Lysinibacillus genus. To measure the residual OTC concentration, a high-performance liquid chromatography method was used. OTC degradation rates were 2.579 mg L−1d−1 with Lysinibacillus strain 3+I and 1.149 mg L−1d−1 without Lysinibacillus strain 3+I. In the presence of strain 3+I, the half-life significantly reduced to 2.68 days, compared to 6.03 days without strain 3+I. The strain demonstrated 85% removal with the OTC concentration of 10 μg/ml. The influence of pH, temperature, carbon sources, and nitrogen source, which influence degradation, were also investigated. The optimum condition favouring degradation was pH 6 at a temperature of 30°C. In addition, Lysinibacillus sp. strain 3+I's ability to degrade OTC in poultry litter offers a promising approach to treat poultry manure and effluent containing OTC, preventing its contamination in the environment.Oxytetracycline (OTC) which is a broad-spectrum veterinary tetracycline antibiotic is extensively used in poultry farms as a prophylactic, therapeutic, and growth stimulator. Upon administration, unmetabolized OTC is excreted from the animal body through droppings and accumulated in litter in the poultry industry. This study aimed at investigating the OTC degradation potential of an-OTC tolerant bacterial strain, isolated from poultry manure. The isolated strain's morphology, biochemical properties, and 16S ribosomal RNA (rRNA) gene sequence confirmed that it belonged to the  Corcoran et al. [B. cereus has been established by Dharmadasa et al. [Acinetobacter, Stenotrophomonas maltophilia, and Aeromonas veronii to OTC in freshwaters of Ireland has been reported by Ding et al. [Ochrobactrum sp. KSS10 [Lysinibacillus 3+I from poultry manure could remove OTC from aqueous solutions.Oxytetracycline (OTC) is a broad-spectrum antibiotic that is used in livestock, particularly poultry, due to its high rate range of application in livestock . For gro0 mg·g−1 . It is a0 mg·g−1 . Current0 mg·g−1 , precipi0 mg·g−1 , irradia0 mg·g−1 , hydroly0 mg·g−1 , and bio0 mg·g−1 . Compost0 mg·g−1 , 33. Com0 mg·g−1 . Most of0 mg·g−1  reportedn et al.  studied n et al.  proved aa et al. , with 56a et al. . The tolg et al. . A reducp. KSS10 . This stPoultry manure was collected from the piled-up manures of five poultry farms randomly located at Coimbatore, Tamil Nadu, India. During August 2019, poultry litter per farm per week was collected using a shovel. We collected manure samples and stored them at −20°C in sterile vials. Homogenised samples were used for the experiments.2HPO4 (0.5 g), and NaH2PO4 (0.5 g) adjusted to pH 7.0. Growth media—nutrient broth and nutrient agar, Mueller–Hinton agar, and OTC (g/l) constituted of (NH4)2SO4 (2.0 g), K2HPO4 (0.5 g), and NaH2PO4 (0.5 g). The solid medium contained 20 grams of agar per litre. Hi-Media provided commercial-grade antibiotic susceptibility discs for six antibiotics: tetracycline, gentamicin, ceftriaxone, oxytetracycline, erythromycin, and vancomycin. Merck provided acetonitrile and methanol of high-performance liquid chromatography grade.For this study, Sigma-Aldrich  provided oxytetracycline of high purity (98%). Inorganic salt medium (ISM) was comprised of—(NH4)2SO4 (2.0 g), Kμg/ml OTC and incubated at 30°C for 48 hours. Using OTC as the sole carbon source, we isolated colonies based on their ability to grow. Using the OTC-ISM agar medium again, the effectiveness of the growth was confirmed at incubation intervals of 0, 1, 2, 3, 5, 7, 14, and 21st days with 100 μg/ml of OTC in ISM broth at 30°C, pH 7.0 and 150 rpm. The growth of the isolated bacteria was determined with a UV-Visible Spectrophotometer . We chose to investigate the OTC-degrading efficiency of the strain showing maximum growth.One gram of poultry manure was transferred to a 250-ml Erlenmeyer flask containing 100 ml of inorganic salt medium (ISM) containing 100 g/ml of OTC. Incubation was carried out at 30°C and 150 rpm/min on a reciprocal shaker for seven days. One ml of this mixture was added to fresh ISM media containing OTC, after every seven days. This was performed four times. The same amount of enrichment culture that exhibited maximum growth with OTC was transferred into sterile test tubes and mixed with 9 ml sterile distilled water, and then serially diluted 10-fold. By using the spread plate technique, 1 ml of the serially diluted culture was inoculated onto OTC-ISM agar plates with 100 According to the bacterial identification manual, the physiological and biochemical properties were determined.Scanning electron microscopy  was employed for the observation of the morphological features of the isolated strain.A series of tests, which included starch hydrolysis, urease utilization, hydrogen sulphide production, tests for catalase, methyl red and carbohydrate fermentation, indole production, oxidase and citrate utilization, and gelatine liquefaction, were carried out .The pure cultures were kept at −20°C in nutritional broth with 15% glycerol and maintained in a refrigerator. Continuous subculturing on ISM agar was performed every 10 days to avoid any contamination and other problems.TM Terminator Cycle Sequencing Kit. In the National Centre of Biotechnology Information (NCBI), the results of the nucleotide sequence analysis were submitted to BLAST analysis (https://blast.ncbi.nlm.nih.gov) in order to generate a universal accession number. Molecular Evolutionary Genetics Analysis Tool (MEGA-X) was used to construct a phylogenetic tree using query and control sequences.As described in our previous study , we perfUtilizing MEGA-X , a phylogenetic tree was constructed from queries and control sequences by the neighbor-joining method, which is based on the Jukes–Cantor model. A neighbor-joining method was used to infer the evolutionary history . BootstrThe isolate was subjected to the antibiotic susceptibility test following the disc diffusion method , 49 for μg/ ml), pH (5–9) temperatures of 20, 30, 35, 40, and 45°C, nitrogen sources such as KNO3, NH4NO3, NH4Cl, and NaNO3, and carbon sources including sucrose, glucose, starch, and fructose.The potential of the strain 3+I to degrade OTC along with their corresponding cell growth was estimated over a period of 7 days. Timed sampling and dilution counting was employed to calculate OTC removal efficiency and cell growth. To measure growth, we quantified colony-forming units (CFU) on agar plates, and residual OTC concentrations were measured using high-performance liquid chromatography. We developed the isolate's dynamic curve by mixing 100 ml of ISM with 10 g/ml OTC at a pH of 7.0 with 3% precultured bacteria and then inoculated the flasks at 30°C and 150 rpm on a shaker. To evaluate the optimum conditions for the degradation of OTC, experiments were performed by varying the initial concentrations of OTC  in the ratio 9 : 1 and 2 : 3 ratios were employed as mobile phases to elute OTC. The mobile phases were sonicated for 5 minutes prior to use. For HPLC determination, aliquots of 0.5 ml were taken from the broth culture on days 0, 1, 3, 5, and 7, and transferred to a sterile 1.5-mL microcentrifuge tube. 0.5 mL extracting solvent comprised of 1 N HCl + 100% acetone + 2 g/L Nae filter  into a s∗ SD) of the results (n = 3). An unpaired t-test was used to analyze changes in the rate of OTC degradation with and without Lysinibacillus sp. 3+I. DMRT was performed with SPSS (version 25) to optimize parameters based on the data.This study used Microsoft Office Excel 2010  for data processing. The error bars in the figures indicate the mean and standard deviation (mean μg/ml OTC agar medium. The isolate OTC-6 showed a higher growth (8.46 ± 0.02 log CFU/ml) with OTC and hence selected for further study. This isolate was labelled as 3+I for experimental purposes. The isolated strain was identified as rod-shaped, Gram-positive bacteria that could form spores and was creamy white in colour as observed from the morphological and biochemical analysis. The scanning electron micrograph (SEM) of Lysinibacillus 3+I revealed the cells as rod-shaped and smooth  and deposited in the GenBank database (MN435620) for universal accession. Using Molecular Evolutionary Genetics Analysis Tool (MEGA-X), a phylogenetic tree was constructed using query and control sequences. BLAST analysis of the 16S rRNA sequence of the isolate exhibited close similarity to the genus Lysinibacillus, based on its biochemical and morphological characteristics.Both strands of the 16S rRNA gene sequence of the isolate were sequenced using PCR techniques using 16S rRNA universal primers . In the The susceptibility tests performed using various antibiotics such as gentamicin, tetracycline, erythromycin, ceftriaxone, vancomycin, and oxytetracycline showed resistance of the isolate 3+I to the studied antibiotics. The zone of inhibition was measured to be 7.6, 8.2, 7.9, 7.3, 7.6, and 8.7, respectively. In accordance with Vlkova et al. , zones oμg/ml of OTC as initial concentration. The cell count increased with an increase in the incubation time, ranging from 7.0 to 8.61 log CFU/ml , denoting that strain Lysinibacillus sp. 3+I had an impact on the reduction of OTC from the medium. In the presence of strain 3+I, 2.68 days was recorded as half-life for OTC in the medium, which is significantly less than the control of 6.03 days suggesting such that microbial degradation is superior to abiotic processes.Using first-order kinetics, the degradation data showed a significant reduction in the OTC concentration with increasing days, with an μg/ml, and the degradation efficiency was highest (92 percent) with 5 μg/ml OTC concentration. The cell growth recorded was higher at 10 μg/ml recording 8.47 CFU/ml and was lower at 5 μg/ml, which was 8.28 CFU/ml , bacterial cells utilize the substrate at a breakaway time. When the concentration of the substrate is depleted (>90%), the bacterial cells die due to the non-availability of substrate. Hence, the viable cell count (CFU) is less on the 7th day. At a higher concentration, the substrate is still available on the 7th day, and hence, the number of viable cells (CFU) is at higher level. The death of bacterial cells has not commenced yet. If the incubation is prolonged further and the degradation reaches >90%, the growth of the organism would have gradually declined due to want of substrate and the number of CFU will come down. When the temperature was raised from 20°C to 45°C, the average degradation was 68, 89, 75, 75, and 70%, respectively  when compared to the abiotic control (54%)   indicatiLysinibacillus. In seven days, strain 3+I was capable of removing 85 percent of OTC, according to the findings. The growth curve and degradation dynamics demonstrated a negative correlation, where an increase in cell growth and time resulted in a drop in the OTC concentration. A positive correlation existed between cell biomass and degradation efficiency. Our results are in par with the findings of Li et al. [β-estradiol by strain E2S. Several species of bacteria have been reported to be tolerant to OTC—Acinetobacter, Stenotrophomonas maltophilia, and Aeromonas veronii [A. nicotianae strain OTC-16 from aqueous media as well as from manure samples. Abiotic control experiments in the absence of strain 3+I showed 54% removal of OTC, which was significantly less when compared to the experiments in the presence of strain 3+I (85%). As reported by Shi et al. [μg/ml OTC (89%) when compared to 5 μg/ml OTC (92%), which may be because of the less availability of OTC to the organisms as has been reported by Ruan et al. [Ochrobactrum sp. decreased its degradation efficiency as temperature increased, as reported by Shao et al. [In the present investigation, the strain 3+I that showed good tolerance to OTC was isolated out of ten bacterial strains from poultry manure. Biochemical characterization revealed that the strain was Gram-negative, motile, rod-shaped bacteria with an ability to form spores and the colony was circular oat meal in colour with smooth edges . The 16S veronii , and fla veronii  which sh veronii  have rep veronii  reportedi et al.  and Harrn et al. . It has n et al. . When pHn et al. . OTC degn et al. . Ochrobao et al. . As the Lysinibacillus 3+I can be considered to share a similar degradation process. The study shows Lysinibacillus 3+I, which is able to utilize OTC for its carbon requirement and operating at pH 6 and 30°C as an appropriate candidate for removing OTC from poultry manure, thus reducing the additional expenditure of adding supplement nutrients to provide optimum conditions.The ability of bacteria to effectively degrade OTC is said to be associated with enzyme productions such as manganese peroxidase , laccaseLysinibacillus 3+I isolated from poultry manure could significantly remove OTC. This organism can be suggested as a suitable candidate for in-depth research and further for incorporation in poultry wastewater pretreatment.To restore our ecosystem and maintain the global carbon cycle established through bioremediation, microorganisms play a critical role. Based on the results of the current study, it is understood that microbes have the ability to clean up contaminants in the environment and could be used as a green instrument for the removal of environmental pollutants. OTC-degrading bacteria have demonstrated their ability to contribute to efforts to mitigate OTC's environmental and public health hazards, and as such, they should be the subject of future research. Our results shows that


Scopolamine is an alkaloid which acts as competitive antagonists to acetylcholine at central and peripheral muscarinic receptors. We report the case of a 41-year-old male convict with a 27-year history of cannabis abuse who suddenly died in the bed of his cell after having smoked buscopan® tablets. Since both abuse of substances and recent physical assaults had been reported, we opted for a comprehensive approach (post-mortem computed tomography CT (PMCT), full forensic autopsy, and toxicology testing) to determine which was the cause of the death. Virtopsy found significant cerebral edema and lungs edema that were confirmed at the autopsy and at the histopathological examination. Scopolamine was detected in peripheral blood at the toxic concentration of 14 ng/mL in blood and at 263 ng/mL in urine, and scopolamine butyl bromide at 17 ng/mL in blood and 90 ng/mL in urine. Quetiapine, mirtazapine, lorazepam, diazepam, and metabolites and valproate were also detected (at therapeutic concentrations). Inmates, especially when they have a history of drug abuse, are at risk to use any substance they can find for recreational purposes. In prisons, active surveillance on the management and assumption of prescribed drugs could avoid fatal acute intoxication. Solanaceae family such as Datura species, Hyoscyamus, and Atropa belladonna. Tropane Solanaceae alkaloids are contained mainly in the seeds, flowers, and leaves. They have strong anticholinergic activity and act as competitive antagonists to acetylcholine at central and peripheral muscarinic receptors. They penetrate the blood–brain barrier and bind to acetylcholine receptors in the cortex and subcortical regions of the brain. At low doses, tropane alkaloids cause peripheral anticholinergic signs including decreased salivation and sweating, mydriasis, loss of accommodation, increased pulse rate, and sinus tachycardia. As the dose increases, ileus, urinary retention, and elevated temperature occur followed by the beginning of central nervous system (CNS) effects . High doses of belladonna alkaloids produce delirium, hallucination, and coma, with higher activity of scopolamine compared to atropine [Scopolamine is one of the main alkaloids, together with atropine, contained in many plants from the atropine .Buscopan® is a pharmaceutical product containing the quaternary ammonium derivative scopolamine N-butylbromide. It acts primarily on parasympathetic ganglia in the walls of the viscera, with antispasmodic action on smooth muscle in the gastrointestinal, biliary, and urinary tracts. It is indicated in the symptomatic treatment of spastic-painful manifestations of the gastroenteric and genitourinary tract and is produced as tablets, suppositories, and injectable solutions . OverdosThe formation of scopolamine from its N-butyl bromide after treatment at high temperature via microwaves or after smoking has been described , 4. The We report the case of a male convict who suddenly died in the bed of his cell. Since both abuse of substances (buscopan® tablets smoking) and recent physical assaults had been reported, we opted for a comprehensive approach (post-mortem CT (PMCT), full forensic autopsy, and toxicology testing) to determine which was the cause of the death.Fig. Fig. A 41-year-old male convict  with a 27-year history of cannabis abuse was found dead in the bed of his cell. Other inmates reported that he had recently smoked an undefined number of buscopan® tablets, obtained by a cellmate. Depakin® , seroquel® (quetiapine), and remeron® (mirtazapine) tablets were found on the floor of his cell. Four days and the day before his death, he had been assaulted, suffering multiple bruises on face, chest, epigastric area and lower limbs, a wound above the left eyebrow, and fractures of six left ribs (from the second to the seventh), and seven right ribs (from the second to the eight). A public prosecutor requested a full forensic autopsy to evaluate whether the death had been caused by the assaults or by a drug intoxication. We performed a CT scan using a Somatom Sensation 16 CT scanner . We adopted these parameters: 140 kVp, 160 mAs, 24-mm feed/rotation, 1-mm slice collimation, 1-mm slice width, and 10–30-40–70-80 reconstruction kernel. After data processing, axial, coronal and sagittal two- dimensional (2D) reconstructions and a 3D-reconstruction with volume rendering (VR) and shaded surface display (SSD) were performed. Virtopsy found significant cerebral edema , and ultrapure water were supplied by Sigma-Aldrich . Scopolamine butyl bromide and sodium valproate were obtained by Sanofi  as 20 and 200 mg/mL solutions, respectively. Zolpidem-D6  was purchased from Lipomed AG  as 0.1 µg/mL solution. One µg/mL solutions were prepared and used as working solutions to prepare the calibration curves.Urine samples were analyzed by LC–MS/MS using a dilute and shoot approach. One hundred microliter of urine were diluted with 400 μL of 0.1% aqueous formic acid, added with a mixture of deuterated internal standards (0.1 µg/mL) and directly injected in the UHPLC-MS/MS system. For the analysis of benzodiazepines another aliquot of sample prepared as described above was preliminary submitted to enzymatic hydrolysis by β-glucuronidase in acidic medium.Peripheral blood samples (0.5 mL) were analyzed and quantified by LC–MS/MS after DLLME (dispersive liquid/liquid microextraction) or after deproteinisation with 0.5 mL of methanol and ultracentrifugation (for scopolamine butylbromide). Detailed analytical conditions for the analysis of blood samples and methods validations are reported elsewhere , 8.Valproic acid determination was performed by GC–MS after liquid/liquid extraction of 2 mL of samples in acidic conditions with ethyl acetate. The extracts were collected, evaporated to dryness, and derivatized with BSTFA (50 μL). One microliter of the extract was injected into the GC port. Calibration curve was in the range 10–100 g/L.2O 0.1% HCOOH/0.1% ammonium formate and B: MeOH 0.1% HCOOH, with a gradient from 10 to 90% B in 14 min. MRM transitions for the detected drugs were the following: Scopolamine 304 156, 138, 103; Scopolamine butyl bromide 360 194, 138; Quetiapine 384 279, 253, 221; Diazepam 285 257, 222, 154; Nordiazepam 271 208, 165, 140; OH-triazolam 359 331, 239, 176; Triazolam 343 315, 308, 239; Oxazepam 287 269, 242, 163; Lorazepam 321 275, 229, 163 Temazepam 301 255, 177, 199; Mirtazapine 266 209, 195, 72.The UHPLC instrument was an Agilent 1290 Infinity system: binary pump with integrated vacuum degasser, high-performance well-plate autosampler, and thermostated column compartment modules. The detection system was an Agilent 6460 triple quadrupole mass spectrometer  with a Jet-Stream electrospray ionization source in positive mode. Instrumental parameters were set as follows: gas temp 350 °C, gas flow 9 mL/min, nebulizer 40 psi, column flow 0.4 mL/min, sheath gas heater 400 °C, sheath gas flow 11 psi, and capillary voltage 4000 V. Chromatographic separation was achieved using a superficially porous Raptor C18 column , mobile phase A: H235, 263; Alprazolam D5 314 286, 210; Diazepam D5 290 154, 227; Nordiazepam D5 276 140, 165; Triazolam D4 347 312, 319; Oxazepam D5 292 246, 109 Lorazepam D4 325 279, 233. Underlined ions were used for quantitation.MRM transitions for the deuterated standards were the following: Zolpidem D6 314 The limits of detection were 0.2 ng/mL for scopolamine and 1 ng/mL for scopolamine butyl bromide, and the limits of quantitation (LOQ) were 0.5 ng/mL for scopolamine and 2 ng/mL for scopolamine butyl bromide. The method was linear in the range from the LOQ to 200 ng/mL for both the substances, giving correlation coefficients of the curves > 0.99 for scopolamine and > 0.98 for scopolamine butyl bromide. The accuracy, intended as %E, was < 15% for scopolamine at 2 and 50 ng/mL and < 20% at 2 ng/mL and < 15% at 50 ng/mL for scopolamine butyl bromide. The validation parameters for the other analytes detected are reported elsewhere [m/z174, 201, 191, and 145 for valproic acid and m/z206 and 233 for GHB-D6, I.S.).The GC–MS system was an Agilent 7890 gas chromatograph coupled to an Agilent 5975c quadrupole mass detector  operating at 70 eV in electron ionization mode. The chromatographic conditions were the following: J&W 5% phenyl-methylsilicone capillary column . Helium was used as carrier gas at a constant flow of 1 mL/min. The oven temperature was held at 75 °C for 2 min, increased to 270 °C at 15 °C/min, and increased to 310 °C at 50 °C/min (held for 4 min). The injection port was set at 270 °C in splitless mode. The mass detector was operated in SIM mode for valproic acid-TMS determination , antipsychotic drugs (quetiapine), and antidepressant drugs . All of them were in therapeutic or subtherapeutic concentrations. The analytical findings are summarized in Table We report the case of a convict who suddenly died in his cell. Since a traumatic cause of death was suspected because of two recent assaults, we performed a PMCT and then a full forensic autopsy, as recommended by the scientific literature , 10. Botolanaceae family, such as Datura stramonium, Atropa belladonna, or Mandragora officinalis. This way of administration causes intoxications with hallucinations, which are the desired effect. Severe complications are often associated with this habit, as anticholinergic and psychotic-like symptoms, but they rarely have a lethal outcome. Cases of paralysis and convulsions following ingestion of flowers of plants of the genus Datura are reported [Datura stramonium seeds or wheat contaminated with seeds were reported. In cases of severe non-fatal intoxications, blood concentrations ranging from 0.5 to 0.8 ng/mL 3 h after the intake were reported; in one case of hypoxemia and coma, blood and urine concentrations were 1.2 and 132 ng/mL, respectively [Scopolamine and atropine intake are generally performed by ingestion of an infusion of the leaves or of the seeds of some plants of the Sreported , 12, 13.ectively . In non-Unlike other cases reported in the literature, in which the way of administration was ingestion, in the present case, scopolamine was administered by smoking crushed buscopan® tablets. The pyrolytic transformation of scopolamine N-butylbromide into scopolamine and its inhalation presumably caused the rapid absorption of a toxic amount of the drug, compatible with a lethal intoxication, as for the high concentration of scopolamine in peripheral blood 14 ng/mL) and in urine (263 ng/mL). After therapeutic use, scopolamine concentrations are in fact reported between 0.1 and 0.3 ng/mL while are considered toxic/lethal concentrations from 1.2 ng/mL . Neverth ng/mL anIn the described case, the presence of other central acting drugs such as valproate, benzodiazepines, mirtazapine, and quetiapine, although in therapeutic concentrations, may have contributed to the fatal outcome by a central depression and alteration of the cardiac rhythm.efficacious, safe, and cost-effective medicines for priority conditions [The misuse of an easily available over the counter drug, considered as essential medicine by WHO, being one of the nditions , must benditions , 17.In this paper, we report the death of a convict caused by an acute intoxication of scopolamine in association to valproate, benzodiazepines, mirtazapine, and quetiapine (in therapeutic concentrations). Inmates, especially when they have a history of drug abuse, are at risk of using any substance they can find for recreational purposes . Therefo


Little is known about the electrophysiological basis of the effect of threat-related emotional stimuli with different motivational direction on duration perception. Thus, event-related potentials were employed to examine the effects of angry expressions and fearful expressions on perception of different duration (490–910 ms). Behavioral results showed there was a greater underestimation of the duration of angry expressions (approach-motivated negative stimuli) than fearful expressions , compared with neutral expressions. Event-related potentials results showed that, the area of Contingent Negative Variation (CNV) evoked by angry expression, fearful expression and neutral expression gradually increased. These results indicated that specific electrophysiological mechanisms may underlie the attention effects of angry and fearful expressions on timing. Specifically, compared with neutral expressions, fearful expressions and angry expressions both are likely to distract more attentional resources from timer, in particular, angry expressions attract more attention resources than fearful expressions from timer. The major contribution of the current study is to provide electrophysiological evidences of fear vs. anger divergence in the aspect of time perception and to demonstrate beyond the behavioral level that the categorization of threat-related emotions should be refined so to highlight the adaptability of the human defense system. Timing is a basic ability of human beings that enables us to adapt to the objective environment . When faThe pacemaker-accumulator (PA) model, as one of the classical theoretical models explaining timing process , postulaIt is more and more recognized that threat-related emotions should be examined separately because each discrete emotion may have a specific function and consequently a specific attention-related effect on duration perception . HoweverAccording to previous literature , we analThrough G-power3.1 software  , the totF = 45.468, p < 0.001, η2p = 0.705; mean ± standard deviation: angry = 3.10 ± 0.50, fearful = 2.95 ± 0.45, neutral = 4.21 ± 0.53; angry/fearful vs. neutral: ps < 0.001] in valence, while no significant difference among the arousal scores of the three types of expressions  occurred. Note that the chosen 20 neutral expressions have a generally higher arousal scores in comparison with the rest of the neutral expressions in the CFAPS to prohibit our results from being polluted by difference in arousal among emotional states [there was a sum of 422 neutral expressions in the CFAPS ].The experiment included a learning stage and a testing stage. A white oval was used as the stimulus of “target” duration for the learning phase. During the testing phase, 60 black-and-white photos  consisting of the same number of facial images for males and females from the native Chinese Facial Affective Picture System (CFAPS) were used as the stimulus of “probe” duration and presented at the center of computer monitoring . The valAfter visiting the laboratory, each participant was seated about 70 cm in front of a DELL computer in a special experimental room. The computer  was responsible for the presentation of the experimental stimuli and recording the participants’ response data with E-Prime 2.0 . Each stimulus was the same as each other in contrast and brightness  against a black background. A temporal generalization task was employed to explore the timing mechanism ,b. The tRecordings were made with a sampling frequency of 500 Hz, an amplifier bandpass of 0.05–80 Hz (6 dB/octave), and an electrode impedance 5 kΩ. Eye movements were subtracted . Tin elepost hoc analysis was performed on behavioral data with affective category  and probe duration  as factors. The significance level was set at 0.05. Where appropriate, the Greenhouse-Geisser correction was applied to ANOVA tests. Post hoc testing of significant main effects was conducted using Bonferroni method. A simple effects model was employed to explain the significant interactions. Partial eta-squared (η2p) was recorded to prove the effect size in ANOVA tests, with 0.0099 showing a small effect, 0.0588 displaying a medium effect, and 0.1379 indexing a large effect  was used for statistical analysis. Mean ± standard deviation are represented as descriptive data. A repeated measures ANOVA with a e effect . For asse effect .‘F = 34.240; p < 0.001; η2p = 0.541], meaning that the temporal distortion induced by two threat-related emotional stimuli were dependent on the probe duration. Simple effect analysis showed that the proportion of “same” responses was modulated by emotion in 490–, 595–, 700–, 805–, 910– ms. In particular, 46.2% angry, 50.6% fearful, and 56.0% neutral expressions under 595 ms condition were estimated as having a duration of 700 ms, while 60.8% angry, 58.7% fearful, and 53.3% neutral expressions under 805 ms condition were estimated as having a duration of 700 ms. The pattern proportion of “yes” responses showed that, compared with neutral expression, the duration of fearful expression and angry expression were estimated to be shorter. Morever, a main effect of probe duration was significant , indicating that the proportion of “yes” responses was smaller in the 490 ms (M = 0.403 ± 0.021) and 910 ms conditions (M = 0.461 ± 0.023), compared to the 595 ms (M = 0.510 ± 0.026), 700 ms (M = 0.553 ± 0.032), and 805 ms conditions (M = 0.576 ± 0.034) (ps < 0.001). The other main effect of emotional expression was present . Post hoc analyses revealed that, compared with the duration of neutral expression (M = 0.519 ± 0.025), the duration of angry expression (M = 0.483 ± 0.025) was more underestimation than that of fear expression (M = 0.500 ± 0.023).A 3  × 5  repeated-measures ANOVA was used to analyze the proportion of “yes” responses . The intF = 146.630; p <0.001; η2p = 0.835)]. Post hoc analyses revealed that, the RT went down with probe duration  going on. Both the effect of discrete emotion  and the interaction effect of discrete emotion by probe duration failed to reach significance .A 3  × 5  repeated-measures ANOVA was used to analyze the response time . There wF = 442.244, p < 0.001, η2p = 0.938], indicating that the VPP amplitude induced by neutral expressions (M = 4.223 ± 0.035 μV), fearful expressions (M = 4.570 ± 0.039 μV), and angry expressions (M = 5.035 ± 0.044 μV) gradually increased significantly (ps < 0.001). Morever, the main effect of the hemisphere also was significant , indicating that the average amplitudes of the VPP in the left hemisphere (M = 4.577 ± 0.040 μV) were significantly smaller than the right hemisphere (M = 4.642 ± 0.041 μV). However, the other effects were not significant  × 5  × 2  repeated-measures ANOVA was employed to analyze the average amplitudes of the VPP component. There was a significant main effect of discrete emotion was present, , indicating that the N170 amplitude induced by neutral expressions (M = -3.690 ± 0.032 μV), fearful expressions (M = -3.942 ± 0.036 μV), and angry expressions (M = -4.645 ± 0.042 μV) gradually increased significantly (ps < 0.001). Morever, The main effect of the hemisphere was significant , indicating that the average amplitudes of the N170 in the left hemisphere (M = -3.568 ± 0.030 μV) were significantly smaller than the right hemisphere (M = -4.617 ± 0.043 μV). However, the other effect were not significant  × 5  × 2  repeated-measures ANOVA was used to analyze the average amplitudes of N170 component. There was a significant main effect of discrete emotion was present, , indicating the CNV area that induced by neutral expressions (M = 3.006 ± 0.028 μV⋅s), fearful expressions (M = 2.634 ± 0.025 μV⋅s), and angry expressions (M = 2.236 ± 0.022 μV⋅s) gradually decreased significantly (ps < 0.001). The main effect of the hemisphere was also significant , indicating that the average amplitudes of the CNV area in the left hemisphere (M = 2.190 μV⋅s) were significantly smaller than the right hemisphere (M = 3.060 ± 0.029 μ V⋅s). In addition, The main effect of probe duration was also significant, , with post hoc analyses indicating that the CNV area for 490 ms (M = 2.070 ± 0.019 μV⋅s) was smaller than for 595 ms (M = 2.414 ± 0.023 μV⋅s); the CNV area for 595 ms was smaller than for 700 ms (M = 2.598 ± 0.026 μV⋅s). However, there was no difference between the area amplitude of CNV at 700, 805 (M = 2.981 ± 0.028 μV⋅s), and 910 ms (M = 3.062 ± 0.031 μV⋅s). The interaction effect of discrete emotion × duration was significant . Simple effect analysis indicated that the effect of emotion on the CNV area first increased, then decreased with the increasing duration of stimulus presentation  in 490-ms condition;  in 595-ms condition;  in 700-ms condition;  in 805 ms condition;  in 910-ms condition  × 5  × 2  repeated-measures ANOVA was used to analyze the average amplitudes of the CNV area. There was a significant main effect of discrete emotion, [ondition .The main aim of the current study was to investigate the effects of two threat-related emotional stimulus, which are arousal-controlled angry expressions and fearful expressions on duration perception with ERPs. Behavioral results revealed that compared with neutral expression, fearful expression and angry expression were being estimated shorter. Several stimulus-locked ERP components were used to objectively characterize the states generated by affective expressions. Accordingly, the ERP data revealed that the early stage of emotion modulated duration perception was represented by larger fronto-central VPP amplitudes and occipito-temporal N170 amplitudes in fear/angry expression conditions, compared with neutral condition. Morever, the CNV, which is an online index of timing, displayed separated waveforms in different emotional stimuli conditions, with smaller amplitudes in angry and fearful conditions when compared with neutral condition.per se relatively low arousing stimuli of fear and anger so as to explore the attention-related mechanism of duration perception. In addition, there might be another explanation of motivational direction influencing the perception of duration  was greater than that in the fear condition (M = 2.634 μV⋅s) and angry condition (M = 2.236 μV⋅s). The medial fronto-central cortex  before the supplementary motor area (SMA) was considered to be the neurosubstrate of the timing function reflected in the CNV  than in the excessive durations (490 and 910 ms). This CNV pattern which showed the duration from 300 to 800 ms was the highest sensitivity on time perception is congruent with previous findings. The main effect of stimulus duration was also significant. The research . The CNVIn summary, arousal-controlled emotional stimuli was used in the current study to verify and compare the effect of two subtypes of threat-related emotional expression on duration perception. We suggested that angry and fearful expressions had different effects on the attention-associated mechanism of time perception, which is issued in divergent behavioral and ERP patterns. Particularly, there was a greater underestimation of the timing of angry expressions than fearful expressions when compared with neutral expressions. ERP results showed that, compared with neutral expressions, a smaller CNV area was observed in the fearful expression condition and angry expression condition, and the trend was more apparent in the angry expression condition. We propose that it is necessary to refine the categorization of threat-related emotions in order to emphasize the adaptability of human defense systems to optimize actions in response to various environmental hazards.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Ethics Committee Southwest Normal University. The patients/participants provided their written informed consent to participate in this study.HY, YB, DL, XC, and JL designed and coordinated the study. YB, GC, LZ, XC, and YO carried out experiments and data process. GC drafted the manuscript. HY reviewed the manuscript. All authors gave the final approval for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Galangin, a natural active bioflavonoid compound, is suggested to inhibit fibrosis and proliferation in certain cells.  Hypertrophic scar (HS) is a benign fibroproliferative skin disease resulting from an aberrant wound healing process and can cause aesthetic and functional damage to patients. Currently, there is no ideal treatment to treat this disease. Galangin could attenuate abnormal scar formation in an HS rabbit ear model. Additionally, the HE staining shows Galangin reduced scar elevation index (SEI) and Masson's trichrome staining changed collagen deposition.  In this study, we found β1 were much lower under a proper dose of Galangin treatment, and Smad7 expression was also enhanced, which are examined by real-time PCR, immunohistochemistry, and western blot.  The expressions of type I collagen, type III collagen, and TGF-Galangin can alleviate dermal scarring via the TGF-β/Smad signaling pathway probably by upregulating Smad 7 expression and, thus, suppressing the expression of type I and type III collagens and TGF-β1. Our data indicated that  Scarring is an inevitable natural process during wound healing. The wound healing progress is a complex one which involves many types of cells, mediators, and three sequential steps: inflammation, new tissue formation, and remodeling , 2. Besiβ (TGF-β) is considered to play the key role [β, activated by Smad proteins in fibroblasts, interacts with TGF-β receptor and leads to an increase in the production of collagen in the ECM and cell proliferation [β/Smad signal path, can prevent receptor-regulated Smads (R-Smads) phosphorylation by integrating with the activated TGF-β1 receptors. Therefore, an overexpression of Smad 7 may possibly attenuate TGF-β1 excretion and inhibit fibrinolysis [However, unlike the scar formation in adults, wound healing of fetal tissue is scarless . The expkey role . TGF-β, feration . Smad 7,inolysis .Galangin is a flavonoid compound extracted from Chinese galangal root and used as herbal treatment in traditional medicine, which is also used in modern clinical treatment because of its antioxidant, anti-inflammatory, and proapoptosis effects [Galangin, we wondered whether it can be used to inhibit HS formation in vivo. Besides, among a large amount of studies to investigate the therapeutic effect of bioflavonoid, little is known about its effect in HS formation. effects –13. Also effects , 15. Conβ/Smad signaling pathway was studied.In this study, HS models were established in rabbits' ears. The effect of bioflavonoid on HS was investigated, and its unique role in regulating the expression of key factors in the TGF-All procedures of this experiment were approved by the Ethics Committee of the First Affiliated Hospital of Xinjiang Medical University. Also, our researchers observed the Animal Welfare Act. Twenty female New Zealand white rabbits  were single-housed under the same standard in the Laboratory Animal Center of Xinjiang Medical University (License No. 20170214-71).A, n = 55) or different doses of Galangin solution , which were purchased from Shanxi Guanchen Biotechnology Co., Ltd., China ) and ultrasonic dissolved with normal saline [On day 0, rabbits were anesthetized by intramuscular injection with 0.15 mL/kg Zoletil 50 (tiletamine hydrochloride and zolazepam hydrochloride) and 0.1 mL/kg ketamine, respectively. Under sterile conditions, six 8 mm diameter round full-thickness dermal wounds were excised down to the bare cartilage on the ventral surface of each ear . The barl saline , 18. Thel saline . On day μm sections, and stained for histopathological analysis. After haematoxylin and eosin (HE) stain, the protuberant degrees were measured as scar elevation index (SEI), which represents the ratio of the scar area to the estimated area below the protuberant portion that had the neogenesis with the same height as the surrounding nonwounded dermis [2O2 incubation for 10 minutes at around 25°C, and then, the sections were incubated with 1% citric acid for 10 minutes in a microwave for antigen retrieval. After that, the sections were blocked with normal goat serum  for 15 minutes at 37°C and then incubated with primary antibodies of collagen I , collagen III , TGF-β1 , and Smad-7  at 4°C overnight. Finally, sections were incubated with goat anti-rabbit secondary antibody  or goat anti-mouse secondary antibody  for 30 minutes at 37°C, followed by DAB  coloration for 5 minutes and haematoxylin staining. All histological images were obtained with an optical microscope connected to cellSens software , and the quantification of positive cells in immunohistochemistry was carried out using Image J software (1.52 v).The 60 HSs (15 in each group) were fixed with formaldehyde, embedded in paraffin, cut into 4 d dermis . To analSixty (15 for each group) HS samples were ground into powders in liquid nitrogen, and then, total RNA was extracted using TRIzol® (Invitrogen) following the manufacturer's instructions. RNA was reverse-transcripted into cDNA with a RevetAid First Strand cDNA Synthesis Kit . Real-time PCR was operated using the Maxima SYBR Green qPCR Master Mix  on the iQCycler thermocycler (Bio-Rad). The primer sequences  are listed in β1 , Smad-7 , and β-actin  overnight at 4°C, respectively. After PBS washing, HRP-conjugated goat anti-rabbit or goat anti-mouse secondary antibody  was added and incubated at around 25°C for 2 hours. The protein bands were visualized using the ECL Kit , and the densities of the bands were compared with that of β-actin as reference.Briefly, 60 HS tissue samples (15 each group) were lysed with Pierce® RIPA buffer  and PMSF (100 : 1). The extracted proteins were measured by using the Pierce® BCA Protein Assay Kit . Proteins were isolated on 12% SDS-PAGE and transferred to polyvinylidene fluoride (PVDF) membranes . Then, the membranes were incubated with primary antibodies of collagen I , collagen III , TGF-t-test. One-way ANOVA was used to compare the mean values of four groups simultaneously, followed by Tukey's test or Tamhane's method while the variance was not homogenous. P value less than 0.05 was considered as statistically significant.All statistic data were analyzed with SPSS version 22 and were demonstreated as mean ± standard deviation (SD). Intergroup comparisons were analyzed by Student's Galangin on scar formation since day 15 after wounding , and type III collagen deposition also decreased from 16.84 ± 0.42 to 13.71 ± 0.47, 11.79 ± 0.60, and 8.83 ± 0.56 . Besides, expressions of TGF-β1 were 15.14 ± 0.71, 17.10 ± 0.36, 12.34 ± 0.73, and 10.88 ± 0.63 , and a higher expression was detected in group B. Also, Smad 7 increased in the Galangin-treated groups, from 4.33 ± 0.26 to 5.47 ± 0.41, 6.81 ± 0.46, and 11.45 ± 0.56 . To further verify the influence of Galangin treatment on the protein expressions of type I collagens, type III collagens, TGF-β1, and Smad 7, western blot was performed. The results demonstrated that type I collagens and type III collagen expressions remarkably reduced in the Galangin-treated groups from 1.32 ± 0.09 to 0.83 ± 0.21, 0.27 ± 0.12, and 0.34 ± 0.16 and from 1.27 ± 0.02 to 0.90 ± 0.09, 0.65 ± 0.11, and 0.34 ± 0.09, respectively . The TGF-β1 protein expressions were 1.13 ± 0.27, 1.56 ± 0.09, 0.36 ± 0.07, and 0.16 ± 0.05, which were effectively suppressed in group C and D, while a higher expression was detected in group B (P < 0.05). By contrast, the Smad 7 protein expression significantly increased after Galangin treatment, from 0.18 ± 0.08 to 0.46 ± 0.25, 0.81 ± 0.13, and 1.21 ± 0.11 (P < 0.05). Thus, it is assumed that Galangin might reduce collagen deposition and TGF-β1 expression, while promoting Smad 7 expression.In order to observe the expression of type I collagens, type III collagens, TGF- group B P < 0.05.β1, and Smad 7, quantitative RT-PCR was carried out. The results demonstrated that the type I collagen mRNA expression was suppressed in all the Galangin-treated groups (P < 0.05), especially in group C . The mRNA expressing of type III collagen and TGF-β1 was inhibited in the Galangin-treated groups, among which the effect in group D  was the most significant . In contrast, the mRNA expression of Smad 7, which is considered as a unique inhibitor of the TGF-β/Smad signal pathway, in the Galangin-treated groups was much higher than that of the control group (P < 0.05). These results suggest that the mRNA levels of type I collagen and type III collagen and TGF-β1 were downregulated after a proper dose of Galangin treatment, while the Smad 7 level was upregulated.To test the mRNA expressions of type I collagens and type III collagens, TGF-Galangin, a kind of bioflavonoid, is found to have antiproliferative property to certain cells [Galangin could suppress HS formation, and this assumption is proved in our further investigation which shows that Galangin could soften the HSs and decrease the SEI of the rabbit ear model in vivo.Scar formation is the normal consequence of skin injury, which is thin and resembles normal skin under correct regulation. However, prolonged inflammation, local mechanical tension, and chronic stimulation may cause excessive deposition of collagen and proliferation of fibroblasts, eventually leading to HS formation , 10, 20.in cells , 23, 24.in cells . Thus, bβ family members, which also participate in regulations of a wide spectrum of cellular functions such as proliferation, differentiation, migration, and apoptosis [β have been found in HS, and the scar-free healing in human fetus is considered as a consequence of TGF-β deficiency [β mediates the fibroblast proliferation, angiogenesis, ECM synthesis, and reepithelialization during the wound healing course [β1 and TGF-β2 was detected in HSs, whereas TGF-β3 was reported to have antifibrotic effects [β1 regulates various fibrosis-related proteins transcriptionally, including type I collagen and type III collagen [β-induced fibrosis in human tubular epithelial cells [β1 protein in group B  might be due to the high dose of bioflavonoid which may cause tissue damage, though no visible wounds were observed on rabbit ears; thus, the proper dose of Galangin in vivo and toxicity study for epidermal cells need further tests. It can also accelerate the transformation of fibroblasts to myofibroblasts, which are considered as major cells in HS formation and characterized as an improved propensity to synthesize collagen and upregulation of cytokines [β1 activities would have potential advantages in inhibiting HS formation.Previous studies have suggested that wound healing and scarring are closely related to TGF-poptosis , 26. Incficiency , 27. Besg course , 29. Ove effects . Particucollagen , 31. A pal cells . Also, oal cells , and theytokines , 34. Theβ receptors and Smads, there may be a greater variety of the signal possibilities than we used to expect. Combinatorial interactions between TGF-β receptors and Smads in oligomeric complexes allow substantial diversity and are complemented by various sequence transcription factors cooperating with Smads, leading to context-dependent transcriptional regulation [β, and it is evidently shown that TGF-β-related protein activation is not a linear signaling transduction pathway. These pathways not only involve Smad-dependent responses but also the Smad-independent responses [β1-activated Smad-dependent pathways can cause cellular changes such as the collagen synthesis and secretion, leading to the aggravated scar formation. However, Smad-independent signaling paths may improve healing. Therefore, inhibition of TGF-β1 transcription may reduce scarring but delay wound healing [Galangin could lessen scarring and delay healing. Meanwhile, the Smad 7 expression was upregulated and that of TGF-β1 was decreased.Despite the limited varieties of TGF-gulation –37. Addiesponses . On the  healing –41. In tR-Smads), common-partner Smads (Co-Smads), and inhibitory Smads (I-Smads). Smad 7, which acts as the I-Smads of TGF-β family signaling, can bind with TGF-β receptor, thus preventing the recruitment and phosphorylation of effector Smads and inhibiting TGF-β/Smads signaling [β/Smad signal path is an autoinhibitory loop to control the intensity and duration of TGF-β signaling response. TGF-β stimulation could cause the export of Smad 7 from the nucleus [β transcriptional responses. Cells with a higher level of Smad 7 are more inclined to resist the profibrotic activities of TGF-β. As this study indicated, Galangin treatment blocked the TGF-β/Smad signaling via increasing the expression of Smad 7, and thus, the stimulation effect of TGF-β1 on collagen deposition in ECM was inhibited.There are three distinct Smad subfamilies: receptor-regulated Smads (ignaling , 38, 42. nucleus . TherefoGalangin could negatively regulate the expression of type I collagens and type III collagens and the collagen deposition. TGF-β1 transcription was downregulated, which might be partially due to Smad 7 upregulation. This study suggests that Galangin may have potential to prevent HS formation.The rabbit ear HS model was established successfully. A proper dose of


Lateral tibial plateau fractures (TPFs) are often treated with conventional open reduction and internal fixation (ORIF) through standard anterolateral sub-meniscal arthrotomy. There has been increasing support for “bidirectional rapid redactor” device-assisted closed reduction and internal fixation (CRIF) for treating TPFs. The aim of the present study is to compare the clinical and radiological outcomes between CRIF and ORIF procedures.We performed a retrospective cohort study of 55 lateral TPF patients (Schatzker types I–III) who accepted surgical treatment at our trauma level 1 center between January 2016 and January 2018. They were divided into the CRIF group (32 patients) and the ORIF group (23 patients) based upon the different surgical protocols. The patients’ clinical outcome analysis was evaluated by using the Knee Society Score (KSS) and Rasmussen’s clinical score. For radiological assessment, changes in tibial plateau width (TPW), articular depression depth (ADD), medial proximal tibial angle (MPTA), and posterior tibial slope angle (PTSA) were evaluated using radiographs and computed tomography (CT) scan.p>0.05). Furthermore, there was no statistically significant difference in terms of age, gender, injury mechanism, follow-up time, time interval from injury to surgery, and Schatzker classification in the two groups. With respect to the clinical outcomes including the KSS score and Rasmussen’s clinical score, there was also no significant difference (p>0.05). Nevertheless, the CRIF group had lower intra-operative blood loss, shorter hospitalization days, and better range of movement of the knee joint than the ORIF group (p<0.05). Furthermore, CRIF had better radiological results when compared to the ORIF group using Rasmussen’s radiological score (p<0.05), although no significant difference was observed in TPW, ADD, MPTA, and PTSA between the two groups (p>0.05).The CRIF group had a mean follow-up of 28.9 months, and the ORIF group had a mean follow-up of 30.7 months (The present study showed that CRIF could achieve comparable clinical outcomes and better radiological results for treating lateral TPFs as compared with conventional ORIF. Tibial plateau fractures (TPFs), accounting for approximately 1.6% of all adult fractures in China , 3. MuchGenerally, the traditional ORIF technique requires excessive soft tissue dissection and may increase the risk of complication rates such as wound infection, neurovascular injury, thrombosis, and soft tissue injuries –11. In r“Bidirectional traction device”-assisted CRIF for TPFs has been successfully applied in the treatment of bicondylar TPFs (Shcatzker V–IV) with advantages of less trauma, lower complication rates, and anatomical restoration of articular congruity . DespiteTo our knowledge, however, the present study is the first to compare the clinical and radiological outcomes between CRIF and ORIF to date. It was our hypothesis that patients with lateral TPFs treated by CRIF had comparable clinical outcomes and better radiological results than ORIF.This study was designed as a retrospective cohort study, and the work complied with the Strengthening the Reporting of Cohort Studies in Surgery (STROCSS) criteria . This stPatients were placed in a supine position with the knees maintained in a flexed 30° position under general anesthesia. A pneumatic tourniquet was used at the proximal thigh for all patients. The surgical technique assisted by “bidirectional traction” device for treating TPFs has been recently reported in our literatures . In the Importantly, in the CRIF group, arthroscopic examination was performed immediately to evaluate the quality of fracture reduction and diagnose and address the concomitant intra-articular structure injuries. For instance, in cases of lateral and/or medial meniscal injuries associated with TPFs, a suture repair or partial meniscectomy was performed simultaneously. In patients with cruciate ligamentous tears, second-stage ligamentous reconstruction would be considered if post-operative knee instability still exists.Patients were evaluated in terms of age, gender, time interval from injury to surgery, mechanism of injury, type of Schatzker fracture, range of movement (ROM), clinical outcomes, and radiological parameters. All patients underwent X-rays and computed tomography (CT) scan of their injured knees pre-operatively. CT scan was performed to evaluate the fracture type according to the Schatzker classification . All theThe Knee Society Score (KSS) and Rasmussen’s clinical score were used to evaluate the patients’ clinical outcomes. Radiological parameters, including tibial plateau width (TPW), articular surface depression depth (ADD), medial proximal tibial angle (MPTA), and posterior tibial slope angle (PTSA), were measured and analyzed from the radiographs and CT scan. The radiological measurements of TPW, ADD, MPTA, and PTSA are shown in Figs. t test was performed for statistical analysis of clinical outcomes and radiological parameters between the two groups, and p value < 0.05 was considered statistically significant for all tests.The SPSS software  was used for the statistical analysis. Continuous variables were recorded as mean ± standard deviation. To determine the number of test patients, the sample size was calculated under a significance level of 0.05 and a power of 0.80. As a result, each group required 20 patients. Categorical variables that presented as percentages were determined by the chi-square test or Fisher exact test. The Student p=0.379), gender (p=0.099), injury mechanism (p=0.315), follow-up time (p=0.187), time interval from injury to surgery (p=0.129), and type of Schatzker fracture (p=0.593) in the two groups (p>0.05). In addition, there were 20 patients associated with intra-articular lesions, of whom 17 in the CRIF group and 3 in the ORIF group, as presented in Table A total of 55 lateral TPF patients treated surgically were enrolled in the present study, and they were divided into the CRIF group and ORIF group, as summarized in Table ism p=0.35, followp=0.027), duration of hospitalization (p=0.003), and range of motion (p=0.014) between the CRIF group and ORIF group (p<0.05). Furthermore, the mean KSS score and Rasmussen clinical score between the two groups were comparable at the final follow-up, and no statistically significant difference was observed between the two groups of patients (p>0.05).The patients’ clinical outcomes are demonstrated in Table p<0.05). One case of infection improved successfully by antibiotic treatment. Three patients with deep venous thrombosis were managed by physical therapy. Furthermore, in one case, stiffness and neural palsy were successfully treated with manipulation under general anesthesia and physical therapy, respectively.The overall complication rate was 10.9% (6/55). It was noted that no complication occurred in the CRIF group while all post-operative complications occurred in the ORIF group, showing a significant difference between the two groups with regard to complication rates (p<0.05). Furthermore, the mean pre-operative TPW was 4.4±3.7 mm in the CRIF group, while that of the ORIF group was 4.3±2.8 mm, being decreased to 1.7±1.5 mm and 1.8±1.6 mm, respectively. Post-operative correction was obvious in both groups, with a statistically significant difference in the width of the tibial plateau (p<0.05). At the final follow-up, the TPW was 2.2±1.2 mm and 2.4±2.1 mm, respectively, showing similar post-operative TPW in the two groups (p>0.05). The mean pre-operative ADD was 12.4±5.7 mm in the CRIF group and 11.8±5.4 mm in the ORIF group, which was significantly decreased to 2.1±1.3 mm and 1.7±1.3 mm after surgery, respectively. At the final follow-up, the ADD was 2.5±1.9 mm and 2.1±1.8 mm, respectively. The mean MPTA was 92.7°±2.3° pre-operatively, 89.2°±1.8° post-operatively, and 88.6°±1.5° at the final follow-up in the CRIF group, while those of the ORIF group were 92.3°±1.9° pre-operatively, 88.7°±2.2° post-operatively, and 87.9°±1.7° at the final follow-up. The mean PTSA was 12.8°±3.7° pre-operatively, 9.6°±3.1° post-operatively, and 10.6°±2.9° at the final follow-up in the CRIF group, while those of the ORIF group were 13.2°±2.8° pre-operatively, 9.8°±4.3° post-operatively, and 10.4°±2.5° at the final follow-up.The radiological results are summarized in Table The most important finding of the present study was that CRIF had comparable clinical outcomes and better radiological results than ORIF. In our study, we identified that the CRIF group had lower intra-operative blood loss, shorter hospitalization days, fewer complication rates, and better ROM of the knee joint, indicating minimally invasive fracture reduction with faster recovery and better functionality when compared to the conventional ORIF technique. However, clinical outcomes were similar concerning the KSS score and Rasmussen’s clinical score, which are in contrast to the findings of a previous study reported by Buckley et al. . They shActually, arthroscopic management in lateral tibial plateau fractures (Schatzker I–III) is widely used and acceptable. Wang et al.  demonstrAlthough the application of arthroscopic examination during surgery increased operation time, our study identified that the CRIF group had no complication related to arthroscopy and had better radiological results with Rasmussen’s radiological score than ORIF alone. This is due in part to the fact that a potential advantage of the arthroscopy may provide a better evaluation for quality reduction and treatment for concomitant intra-articular lesions through a less invasive procedure than ORIF. Furthermore, the bidirectional tractor, a reliable and minimally invasive procedure, provided advantages of nearly anatomical reduction of fracture, restoration of articular congruity, and less dissection of soft tissue. Thus, this reinforces the superiority of arthroscopy management combined with CRIF procedure in treating patients with TPFs, which are consistent with the viewpoint of the Canadian Orthopaedic Trauma Society .The targets of the surgical treatment in TPFs are not only the anatomical reduction of articular congruity and restoration of lower limb alignment, but also the reasonable treatment of concomitant intra-articular injuries to allow early rehabilitation, enhancement of knee stability, and minimize the risk of post-traumatic osteoarthritis. In addition, TPFs are often associated with meniscal tears and cruciate ligamentous injuries. Abdel-Hamid et al.  arthroscSeveral limitations of this present study should also be noted. First, selection bias was inevitable due to the nature of the retrospective study. Second, this study in calculating intra-operative blood loss is not accurate due to the use of arthroscopy with irrigation fluid at the time of fracture fixation. Third, the number of patients was relatively small, and the follow-up time was short, which may not observe the development of post-traumatic osteoarthritis. Fourth, a second-look arthroscopic examination to demonstrate the therapeutic effect of the intra-articular soft tissue injuries in TPFs was absent.Our study showed that CRIF had comparable clinical results with ORIF, and it had advantages over ORIF with regard to better radiological results, less trauma, and fewer complication rates. Furthermore, we recommend that arthroscopic examination should be performed after internal fixation to diagnose and address concomitant intra-articular soft tissue injuries.


The study aimed to estimate the prevalence and correlates of underweight and overweight/obesity among adults in Afghanistan.National cross-sectional survey data of 3779 persons aged 18–69 years were analysed. Multinomial logistic regression was used to estimate factors associated with underweight and overweight/obesity relative to normal weight.2), 49.5% had normal weight (BMI 18.5–24.9 kg/m2), 25.5% overweight (25.0–29.9 kg/m2), and 17.2% obesity. In adjusted multinomial logistic regression, factors negatively associated with underweight were male sex (adjusted relative risk ratio (ARRR): 0.30, 95% confidence interval (CI): 0.15–0.58) and hypertension  and factors positively associated with underweight were sedentary behaviour  and current tobacco use . Factors positively associated with overweight/obesity were aged 30–44 years  and aged 45–69 years  (compared to 18–29 years) , hypertension , and type 2 diabetes , and high physical activity  was negatively associated with overweight/obesity.In all, 7.8% of the study sample was underweight (BMI < 18.5 kg/mAlmost one in ten adult respondents were underweight and more than two in five were overweight/obese, confirming a dual burden of malnutrition in Afghanistan. Afghanistan is a low-income country, its living standards are among the lowest in the world, it has a population of 36.6 million, life expectancy at birth was 52.8 years, and the adult literacy rate was 43% (55.5% among men and 29.8% among women) [In local community surveys in urban centres in Afghanistan, the following proportions of body weight status have previously been shown: in Kabul City , underweight 1.1% and overweight/obesity 69.3% ; in KabuBoth undernutrition and overnutrition in adulthood have been linked to various negative health effects, including morbidity and mortality , 17. As This is a secondary analysis conducted using nationally representative population-based and cross-sectional data from the “2018 Afghanistan STEPS survey” . The 2012 underweight, 18.5–24.4 kg/m2 normal weight, 25–29.9 kg/m2 overweight, and ≥ 30 kg/m2 obesity” [Anthropometric measurements were taken by trained healthcare staff in a safe and secure area; height and weight were measured using a portable electronic weighing scale and measuring inflexible bars . “Heightobesity” .Hypertension or raised blood pressure (BP) was defined as “systolic BP ≥ 140 mm Hg and/or diastolic BP ≥ 90 mmHg or where the participant is currently on antihypertensive medication.” [cation.” . BP was cation.” .Diabetes was defined as “fasting plasma glucose levels ≥ 7.0 mmol/L (126 mg/dl); or using insulin or oral hypoglycaemic drugs; or having a history of diagnosis of diabetes” [iabetes” .Health risk behaviour variables comprised alcohol use, tobacco use status, sedentary behaviour (≥ 8 h/day [ 8 h/day ), and “l 8 h/day .Dietary behaviour included “daily fruit and vegetable consumption measured from the total number of servings of fruit and vegetables eaten per day in a typical week.” Meals outside the home were assessed with the question, “On average, how many meals per week do you eat that were not prepared at home? By meal, I mean breakfast, lunch and dinner?” (Number) [(Number) . Adding (Number) .Sociodemographic information included age, sex, highest educational level, number of adult household members, and residence status [e status . Househoe status .P < 0.05 was considered significant.All statistical procedures were adjusted for complex sample design and conducted with “STATA software version 13.0 ”. The data were weighted “to make the sample representative of the target population in Afghanistan .” . Chi-squ2), 49.5% had normal weight (BMI 18.5–24.9 kg/m2), 25.5% overweight (25.0–29.9 kg/m2), and 17.2% obesity. Further sample details are shown in Table The sample consisted 3779 individuals aged 18–69 years , and 55.9% were male. Two in five participants (41.8%) were living with six or more adult household members, 59.0% had no formal education, and 42.4% lived in rural areas. In all, 7.8% of the study sample was underweight : 0.30, 95% confidence interval (CI): 0.15–0.58) and hypertension , and factors positively associated with underweight were sedentary behaviour  and current tobacco use . Factors positively associated with overweight/obesity were aged 30–44 years  and aged 45–69 years  (compared to 18–29 years) , hypertension , and type 2 diabetes , and high physical activity  was negatively associated with overweight/obesity (see Table 2) in this study is lower than the prevalence rates found in urban centres in Afghanistan, e.g., in Kabul City  [In this national 2018 Afghanistan STEPS survey, the prevalence of underweight 7.8%) was higher than in Kabul City  , in Jala.8% was h11–2012) , in Kabu11–2012) , in Kabu11–2012) , in Iran11–2012) , in Iraq11–2012) , in Moro2) in the low-income country, Afghanistan. The co-existence of undernutrition (15.6%) and overnutrition (18.0%) has also been found in low-income countries in the Asia Pacific region [Findings show the double burden of undernutrition (7.8%) and overnutrition  and among women (9.1%), which was also found in previous studies –33, and In bivariate analysis, obesity was higher in women (23.7%) compared to men 12.1%), which is in line with previous studies .1%, whic, 36. ConIn terms of dietary behaviour, previous research found associations between inadequate fruit and vegetable intake , 40, eatIn agreement with previous studies , 45, 46,Apart from physical and biomedical measures self-reported questionnaire data may have suffered from biased responses. Another limitation was the cross-sectional nature of the survey, which does not allow for causative conclusions. Some variables, such as more details on dietary behaviour, should be included in future studies.The study found in the 2018 adult national Afghanistan STEPS survey that almost one in ten adult participants were underweight and more than two in five were overweight/obese. Several factors associated with body weight status, including female sex, current tobacco use and sedentary behaviour for underweight and older age, hypertension, type 2 diabetes, and physical inactivity for overweight/obesity, were identified. Increased public health interventions are needed to address both forms of malnutrition (underweight and overweight/obesity) and associated factors to improve the health of Afghans.


Cinnamomum zeylanicum stem bark aqueous extract (AECZ) on gentamicin-induced nephrotoxicity. AECZ was prepared by maceration in water and tested orally at the doses of 200 and 400 mg/kg/day to prevent gentamicin-induced nephropathies in male Wistar rats. Gentamicin (100 mg/kg/day) was administered for 14 consecutive days by intraperitoneal route, concomitantly with AECZ or silymarin (50 mg/kg/day) used as reference drug. Animal body weight was monitored during the treatment. After the last treatment on the 14th day, animals were sacrificed. Blood was collected for the evaluation of hematological and renal function biomarkers. The homogenate of one kidney was used to assess oxidative stress markers and proinflammatory cytokines, while the other one was fixed in formaldehyde for histopathological studies. Gentamicin decreased body weight, serum total proteins, and calcium level but increased kidneys' relative weight, serum creatinine, urea, and uric acid. Moreover, the levels of reduced glutathione, catalase, and superoxide dismutase activities were decreased, while an increase in malondialdehyde, proinflammatory cytokines , and nitrites was observed in the negative control group as compared to normal control. Histological analysis of the kidney revealed the presence of tubular necrosis, glomerular degeneration, and macrophage infiltration in the gentamicin-treated group. All these impairment parameters were prevented by AECZ and silymarin treatments. AECZ has a protective effect against gentamicin-induced nephrotoxicity. The antioxidant and anti-inflammatory potentials of this extract may highly contribute to its nephroprotective activity.Nephropathies and especially nephrotoxicity have become one of the serious causes of life-threatening conditions because of intensive exposure to xenobiotic whether by environmental pollution or by drug abuse. The present study was undertaken to assess the protective effects of  Nephrotoxic medications can lead to damage to the kidney via various mechanisms, including structural and functional alterations. When evaluating the primary causes in renal injury, the incidence of drug-induced toxicity has accounted for 20% of all-cause incidents , 2. Manyfunction –7. Severvulgaris , 9. ThisCinnamomum zeylanicum (Lauraceae), also known as true cinnamon, which originates from Sri Lanka and south India, is a tree that is about 10 meters high and can withstand hard climatic conditions. It is found predominantly in tropical and subtropical regions, where the rainfall and the temperature range from 1500 mm to 2500 mm and from 27°C to 30°C, respectively [ectively .C. zeylanicum have demonstrated its therapeutic potentials including antihypertensive and vasorelaxant activities [Cinnamomum zeylanicum stem bark also demonstrated acute and chronic antihypertensive properties in NO-deficient hypertensive rats [Cinnamomum zeylanicum stem barks can attenuate diclofenac sodium and oxytetracycline mediated hepatorenal toxicity by modulating oxidative stress, cell apoptosis, and inflammation in male albino rats [Many communities in Cameroon use the stem bark of this plant traditionally for the management of various ailments such as muscular pain, rheumatism, gastrointestinal disorders, and typhoid fever . It is ativities , antinoctivities , antidiativities , immunomtivities , and anttivities . The metive rats . Recentlino rats . Also, cino rats .Cinnamomum zeylanicum contain 45%∼65% cinnamaldehyde, 12%∼18% eugenol, and small amounts of 2′-hydroxycinnamaldehyde and 2′-benzoyl-oxycinnamaldehyde [The stem barks of aldehyde , 22.Cinnamomum zeylanicum [Cinnamomum zeylanicum are mainly due to the presence of cinnamaldehyde and eugenol [Cinnamaldehyde, a phenolic compound primarily identified as the main phytochemical found in cinnamon bark, is responsible for most of the biological effects of ylanicum . Various eugenol , 25. It  eugenol , 26.Cinnamomum zeylanicum stem barks has been demonstrated [C. zeylanicum protects against kidney damage are not known. The present study was undertaken to determine the preventive effect of AECZ extract in gentamicin-induced nephrotoxicity by investigating the impact of treatment on some renal biomarkers, oxidative stress parameters, proinflammatory cytokines, and histopathological features.Although the nephroprotective activity of the ethanolic extract of nstrated , no infoCinnamomum zeylanicum parts were harvested in Njombe  in August 2018 and used for the authentication of the plant in Yaoundé National Herbarium by comparison with the voucher specimen SRFC/22309. The stem barks were air-dried at room temperature and ground into a fine powder. 800 g of this powder was macerated into 7 L of distilled water for 48 hours. The mixture was then filtered with Whatman filter paper no. 3. The filtrate was lyophilized, and 30.78 g of the aqueous extract was obtained from this preparation (yield of 3.85%).Thirty-six male, 8-9-week-old Wistar rats weighing between 135 and 145 g at the beginning of the experiment were obtained from the animal house of the Department of Animal Biology and Physiology, Faculty of Science, University of Yaoundé 1, Cameroon. They were maintained under standard laboratory conditions with natural light cycle and had free access to normal laboratory rat chow and tap water. The study was conducted in accordance with the guidelines of the Cameroon National Ethical Committee of the use of laboratory animals for scientific research (ref. NFWA-IRD 0001954).p.o.) and saline solution 0.9% , a negative control group which received distilled water  and gentamicin , a positive control group which received silymarin  and gentamicin , the two test groups which received the aqueous extract of Cinnamomum zeylanicum (AECZ) at the doses of 200 mg/kg and 400 mg/kg (p.o.) and gentamicin , and the extract control group which was treated with AECZ  and saline solution 0.9% . The doses of the extract were chosen from literature based on their protective effect against acetaminophen-induced cellular damage and apoptosis in renal tissue [The thirty-six rats were randomized and assigned to six groups composed of six animals each and treated daily for 14 days as follows: a vehicle control group which received distilled water  and blood from the carotid arteries was collected using capillary tubes under an anticoagulant (Ethylenediaminetetraacetate) or not, for hematological and biochemical studies, respectively. After blood collection, the rats were sacrificed by decapitation. The two kidneys were collected and dry-blotted and the relative organ weight was calculated.At the end of the treatment on day 15, animals were fasted overnight but had free access to water. They were then anesthetized with urethane . The parameters considered were the red blood cells (RBC) count, the white blood cells (WBC) count, the hemoglobin (Hb) concentration, the hematocrit (HCT), the mean corpuscular hemoglobin (MCH), the mean corpuscular volume (MCV), the mean corpuscular hemoglobin concentration (MCHC), the platelet count, the lymphocyte, the monocyte, and the granulocyte counts.+, K+, and Cl− were determined using automatic equipment .Serum was separated from the clotted blood sample by centrifugation  and the collected sample was stored at −20°C for analysis. Glucose (CN : BXC0101A), albumin (CN : BXC0221A), aspartate aminotransferase (CN : BXC0128A), urea (CN : BXC0126A), creatinine (CN : BXC0117A), uric acid (CN : BXC0603B), and total calcium (CN : BXC0292A) were assayed in serum samples, using corresponding commercial diagnostic kits purchased from Fortress, UK. Total protein was evaluated using the method of Bradford (1976). Serum NaOne of the collected kidneys was homogenized at 20% (w/v) in Tris-HCl 50 mM buffer (pH 7.4). The obtained homogenates were centrifuged at 1510×g for 15 minutes at 4°C. The supernatant collected was stored at −20°C for further biochemical assays. The total protein was determined by the method of Bradford . The supβ (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), in kidney homogenate were measured using a Sandwich enzyme-linked immunoassay kits specific to each cytokine . The measurements were performed at 450 nm using a microplate-reading spectrophotometer (Thermo Scientific Multiskan FC). The levels of TNF-α, IL-1β, and IL-6 were expressed as pg/mg of protein [The levels of the inflammatory cytokines, interleukin-1 protein .μm were made and further stained with hematoxylin and eosin (H&E). The sections were examined with light microscope (Zeiss Axioscope) and digital camera, and MiniSee software was used for images capture. As an example of kidney damage, tubular clarifications were quantified using ImageJ software, version 1.48 for Windows. The percentage of tubular clarifications was calculated as follows:For histological analysis, the second kidney was fixed in 10% formalin for 7 days and further paraffin-embedded. Sections of 4–5 p < 0.05 was considered statistically significant. All analyses were performed using GraphPad Prism software, version 8.01.Data were expressed as mean ± standard error of mean. Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by the Tukey post hoc test. p < 0.001) decreased the rat weight gain by 45.93% and increased the relative kidney weight by 48.34% as compared to normal control group. AECZ at the doses of 200 and 400 mg/kg significantly (p < 0.001) prevented the decrease of weight gain and the increase of relative kidney weight by 14.13% and 17.39%, respectively, compared to gentamicin group. AECZ administered alone significantly increased the body weight gain as compared to normal control (Gentamicin significantly ( control .p < 0.001) the number of monocytes compared to the normal control. Silymarin (50 mg/kg) and AECZ at the doses of 200 and 400 mg/kg significantly increased the number of monocytes by 52.50% (p < 0.05), 90% (p < 0.001), and 67.50% (p < 0.01), respectively, compared to the group treated only with gentamicin.p < 0.05) and 23.07% (p < 0.01), respectively. In the GM group, the serum concentration of albumin increased significantly by 15.91% (p < 0.001) compared to normal control group, but concomitant treatment with GM and AECZ at the doses of 200 and 400 mg/kg significantly (p < 0.001) reduced the serum albumin by 12.83 and 13.73, respectively. The serum concentration of aspartate aminotransferase increased significantly by 57.7% (p < 0.001) in the GM group when compared to normal control. The administration of C. zeylanicum extract at the doses of 200 and 400 mg/kg or silymarin (50 mg/kg) resulted in a significant reduction in ASAT activity, respectively, by 52.71% (p < 0.01), 64.70% (p < 0.001), and 53.38% (p < 0.01) as compared to gentamicin-intoxicated rats. None of the evaluated parameters were affected by AECZ when administered alone.The effect of AECZ on serum glucose, total proteins, albumin, and aspartate aminotransferase (ASAT) is presented in p < 0.001) inhibited the increase of serum creatinine by 43.72% and 47.16%, urea by 57.08% and 47.16%, and uric acid by 57.08% and 47.16%, respectively. AECZ at the doses of 200 mg/kg and 400 mg/kg also prevented the decrease of calcium level by 122.48% and 116.06%, respectively. Likewise, silymarin also prevented this decrease. The serum levels of potassium and sodium were not significantly affected by any of the treatments as compared to normal control.The effects of AECZ on kidney's function were evaluated by determining the rate of creatinine, uric acid, urea, and electrolytes  in serum . GentamiC. zeylanicum stem barks on some kidney markers of oxidative stress are shown in p < 0.001) by 73.79% (10.3 ± 1.14), compared with the normal control group (39.3 ± 5.05). However, pretreatment with silymarin (50 mg/kg) and AECZ (200 mg/kg and 400 mg/kg) significantly (p < 0.001) enhanced superoxide dismutase concentration by about 300.97%, 406.79%, and 377.66%  compared with the gentamicin group (10.30 ± 1.14).The effects of the aqueous extract of p < 0.001) in the renal activity of CAT (68.60 ± 2.89) as compared to normal control rats (168.00 ± 11.00). Silymarin (50 mg/kg) significantly increased (p < 0.001) the activity of CAT in the kidneys by 76.38% as compared to the GM group (121.00 ± 12.7 versus 68.60 ± 2.89). AECZ at the doses of 200 mg/kg and 400 mg/kg significantly enhanced CAT concentration in a dose-dependent manner by about 57.43% (108.00 ± 3.33) and 86.58% (128.00 ± 7.91) compared to gentamicin group (68.60 ± 2.89).Treatment with GM induced a significant decrease  for 14 days decreased the content of GSH significantly by about 70.87% (43.40 ± 5.43), compared to the normal control group (168.00 ± 11.01). Moreover, pretreatment with silymarin (50 mg/kg) or AECZ 200 mg/kg and 400 mg/kg one hour before gentamicin administration caused significant 2.3-, 3.2-, and 2.94-fold increases in renal glutathione content , respectively, compared with the gentamicin group (43.40 ± 5.43).p < 0.01) of renal lipid peroxidation, which manifested as an increase in malondialdehyde concentration (4.37 ± 0.13), compared with the normal control group (3.19 ± 0.02). Pretreatment with silymarin (50 mg/kg) and AECZ (200 mg/kg and 400 mg/kg) for 14 days before gentamicin significantly reduced malondialdehyde concentration by about 38.58%, 38.21%, and 37.75% reaching 2.64 ± 0.21, 2.70 ± 0.33, and 2.72 ± 0.14, respectively, in comparison with the gentamicin group (4.37 ± 0.13).Gentamicin injection (100 mg/kg/day) for 14 days caused significant elevation (p < 0.01) in kidney protein content as compared to normal control, whereas there was a significant increase in total protein content when animals were treated with silymarin , AECZ 200 mg/kg , and AECZ 400 mg/kg  as compared to GM-treated rats.Gentamicin-treated rats (negative control) showed significant decrease of 27% (p < 0.001) in gentamicin-treated rats as compared to normal rats. This concentration of nitrite was decreased significantly by 40.81% (p < 0.01) in silymarin group, by 53.78% (p < 0.001) in AECZ (200 mg/kg) group, and by 47.02 (p < 0.001) in AECZ (400 mg/kg) group as compared to GM-treated rats.The level of nitrite was increased significantly by 112.64% (p < 0.001) 2.21-fold elevation of TNF-α (116.0 ± 6.3 to 257.0 ± 18.9 pg/mg of protein). However, when these animals were concomitantly treated with silymarin (50 mg/kg) and GM, a 1.43-fold significant (p < 0.001) decrease in the TNF-α levels was noted. In groups treated with AECZ at the doses of 200 and 400 mg/kg, 1.34-fold and 1.70-fold decreases of the TNF-α levels were, respectively, observed.As depicted in p < 0.001) 2.17-fold elevation of IL-6 (from 39.3 ± 1.72 to 85.4 ± 3.45 pg/mg of protein) when compared to normal control rats. As compared to the GM-treated rats, silymarin (50 mg/kg) significantly (p < 0.001) decreased the IL-6 levels by 1.81 folds. In groups treated by AECZ at the doses of 200 and 400 mg/kg, significant (p < 0.001) 1.58-fold and 1.86-fold decreases of the IL-6 level were, respectively, observed.GM administration induced a significant  elevation of IL-1β (from 22.9 ± 1.31 to 51.9 ± 1.6 pg/mg of protein).p < 0.001) decreased the IL-1β levels by 1.37 folds (from 51.9 ± 1.6 to 37.8 ± 1.85 pg/mg of protein). Treatment with AECZ at the doses of 200 and 400 mg/kg significantly decreased the level of IL-1β, respectively, by 1.40 folds (p < 0.01) and 1.66 folds (p < 0.001).As compared to GM group, silymarin (50 mg/kg) significantly  increase of tubular clarification (p < 0.001) this damage , and nitrites compared to gentamicin-treated group. Histological analysis of the kidney revealed that AECZ prevented kidney alterations induced by gentamicin.The purpose of this work was to evaluate the potential nephroprotective activity of the lyophilized aqueous extract of Gentamicin has been intensively used as an experimental model of nephrotoxicity as it perfectly reflects clinical cases. The main aspect of gentamicin nephrotoxicity is tubular cytotoxicity. In fact, it causes tubular damage through necrosis of tubular epithelial cells predominantly in proximal segment and alteration of function of main cellular components involved in transport of water and solutes. In the present study, 14 days of gentamicin injection reduced body weight gain and increased the kidney's relative weight. The decrease of body weight gain could be a result of increased proteolysis and reduced proteins synthesis as previously demonstrated –37. The 2– and NO• generation through activation of inducible isoform of NO• synthase (iNOS) in kidney [• and further boosted the endogenous antioxidant parameters including SOD, catalase, and GSH, suggesting its in vivo antioxidant effects. This antioxidant effect of AECZ might contribute to its nephroprotective activity.Oxidative stress activation and inflammatory process constitute a major part of gentamicin nephrotoxicity. In fact, nephrotoxicity induced by gentamicin is associated with the mitochondrial dysfunction in renal tubular cells. It causes a marked impairment in the activity of mitochondrial respiratory enzymes including NADH dehydrogenase resulting in the excessive ROS generation . Furthern kidney  tissue. n kidney ,  40. AEFor more insight into the beneficial effects of AECZ, we evaluated additional parameters of nephron-cytotoxicity and renal function. Gentamicin induced significant increase in the blood levels of creatinine, urea, uric acid, albumin, and ASAT, indicating renal necrosis and dysfunction , 41. Admα, IL-1β, and IL-6).The development of oxidative stress coupled with the necrosis triggers the inflammatory process , 40. InfCinnamomum zeylanicum stem bark on nociception and anxiety demonstrated that the anti-inflammatory effect is ascribed to trans-cinnamaldehyde component of cinnamon [The role of these proinflammatory mediators is well known in the kidney diseases ,  42. Incinnamon , 45.Any toxic substance that triggers good functioning of the kidney provokes an inflammatory response in the kidney tissue, as well as its hypertrophy , 47; theThe haematopoietic system is one of the most sensitive targets for toxic compounds and an important index of physiological and pathological status in man and animal. Surprisingly, apart from the monocytes whose number decreased in animals treated with gentamicin, no other hematological parameter varied significantly independently of the treatment. This fact remains to be understood.Aminoglycosides cause disturbance of renal functions by accumulating on the membranes of the proximal convoluted tubule, impairing the reabsorption and secretion of electrolytes and in that way causing homeostatic imbalance. In the present study, serum calcium level was reduced significantly and a nonsignificant decrease in serum sodium and potassium levels in gentamicin-treated rats was observed. These observations on serum sodium and potassium levels, although nonsignificant, are in agreement with those of Nafiu et al.  showing Cinnamomum zeylanicum stem bark are cinnamaldehyde and eugenol. It has been well documented that they have various therapeutic properties like antioxidative and anti-inflammatory properties, as well as nephroprotective effects [Evidences have indicated that the renal toxicity of gentamicin is due to its selective accumulation in the renal proximal convoluted tubules, which subsequently leads to the loss of the tubule brush border integrity, severe degeneration, necrosis in epithelial cells of the proximal tubules, and infiltration of mononuclear cells in intertubular spaces , 49. In  effects , 50–52.  effects .Data from the present study evidenced the nephroprotective effect of AECZ against gentamicin toxicity and further support the use of this plant by the local populations in the management of kidney diseases. The nephroprotective effect of AECZ is more likely mediated by its antioxidant and anti-inflammatory capacities. AECZ might be a good candidate for the development of new drugs that can protect against acute kidney diseases.


However, it was observed that plastic products constituted the highest percentage of the pollutants found in almost all the stations, with plastic bags being the most dominant.This study was conducted around the beaches in Indonesia in order to investigate the level of pollution in the marine environment. Thirteen (13) locations in six (6) regions namely: Seribu Island, Banten, South Java, Biawak Islands, West Papua, and East Nusa making up a total length of 12.84 km of the area were studied. This investigation focused on the debris around isolated beaches, tourist attraction centers, fishing zones and marine protected areas (MPA). The method employed in this study was dependent on the international coastal cleanup form. The samples of debris collected and studied varied from the year 2013 to 2018 for a thorough investigation. The beach debris monitoring equipment revealed information about the distribution, abundance, types and, effects of marine debris on the ecosystem. Moreover, the study showed that the mass of debris collected within the areas listed weighted 1113.10 kg for 34,330 collected items. Also, the average density was noted to range between 1.43 and 5.11 items/m However, one of the biggest problems identified in this area is the introduction of debris to the coastal environment.•Debris on the beaches may come from any source, and the different beaches have been identified with different types of debris.1Coastal areas have the potential to collect garbage from two main sources The dataset contained information about macro litter (>2.5 cm) acquired in 13 locations with a total of 121 sample points collected from 2013 to 2018 . The beaDebris was collected from 121 stations in six areas, weighing around 1.1 tons with an average of 8.81 ± 0.08 kilos per 100 m and approximately 34,330 items collected from 12.84 km total length. An accumulated average of marine debris is 260 ± 2.51 items per 100 m of beach length were collected from all the stations. Both plastic and rubber items dominated the pollutants collected in each of the stations. Plastic bags, plastic bottles, foam, tires, and rope are samples of the most likely picked items in this category .Fig. 2Sa2 with the highest varying from 0.01 to 18.71 items/m2. On average, South Java reported the highest marine debris from all observed stations with values of 5.11 Items/m2, and 0.096 kg/m2  of Tunda, Pangandaran, Untung Jawa, Indramayu, Panjang Island, Biawak, Harapan Island, and Handeleum, respectively .Fig. 5Se22.1https://oceanconservancy.org/wp-content/uploads/2017/04/OC-DataCards_volunteerFINAL_ENG.pdf). The line in Transect (LIT) method was used between 100 m parallel to the coastline The sampling method used in this research is based on International Coastal Cleanup (ICC) form developed by Ocean Conservancy International  methods is a technique deployed to test the quality of acquired data. The best method to ensure the acquisition of good quality data is that the method and procedure for data collection should be simulated before sampling https://data.mendeley.com/datasets/r3y43cdd3x/2.The data is stored in Ibnu Faizal: Conceptualization, Writing – original draft; Zuzy Anna: Funding acquisition, Conceptualization; Sanny T. Utami: Validation, Writing – review & editing; Putri G. Mulyani: Data curation; Noir P. Purba: Writing – original draft, Methodology.The authors declare that there are no known competing financial interests or personal relationships that could have influenced the work reported in this paper.


Using this assay, reproducible MICs were obtained for amphotericin B, itraconazole, voriconazole, posaconazole, terbinafine, and micafungin.We present an  in vitro susceptibility assay for Madurella mycetomatis hyphae using resazurin for endpoint reading. Using this assay, reproducible MICs were obtained for amphotericin B, itraconazole, voriconazole, posaconazole, terbinafine, and micafungin. Results were comparable with those of a 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT)-based susceptibility assay. The lowest MICs were obtained for itraconazole and posaconazole  followed by voriconazole . Amphotericin B, micafungin, and terbinafine appeared much less effective.We present an  Madurella mycetomatis is the most common causative agent of eumycetoma. Unfortunately, none of the antifungals currently available for use seem to be therapeutically effective in eumycetoma caused by M. mycetomatis infection. Antifungal therapy always needs to be combined with drastic surgical procedures -2H-tetrazolium-5-carboxanilide salt (XTT) was used as a viability dye to quantify growth after 7 days of incubation , triazoles , echinocandins , and allylamines  was determined. Resazurin-based results were compared with those obtained with the XTT assay published previously . Concentration ranges were as follows: 0.125 to 8 µg/ml for amphotericin B; 0.016 to 1 µg/ml for itraconazole, posaconazole, and voriconazole; 2 to 128 µg/ml for micafungin; and 0.5 to 32 µg/ml for terbinafine. An M. mycetomatis inoculum was prepared in RPMI 1640 medium containing 0.35 g/liter l-glutamine and 1.98 mM 4-morpholinepropane sulfonic acid . After 7 days incubation at 37°C, the mycelia were centrifuged for 5 min at 2,628 × g . The medium was poured off, and fresh medium was added. The mycelia were again sonicated for 10 seconds at 28 µm maximum power and adjusted to transmission of 35% to 45% at 660 nm . A 100-µl standardized hyphal suspension was added to each well of a round-bottom 96-well plate  along with 1 µl of the antifungal agent and 1 µl of resazurin solution (0.15 mg/ml). A growth control and a negative control were included. After being added, the readily soluble resazurin formed a deep blue solution. The plates were then sealed and incubated at 37°C for 7 days. During growth, resazurin is transformed into pink fluorescent resorufin in viable metabolically bioactive fungal cells. The quantity of resorufin produced is proportional to the number of viable cells and can be assessed spectrophotometrically or visually without the use of specialized equipment (600 NC − OD600 test/OD600 NC − OD600 GC) × 100, and percent growth inhibition = 100 − (OD450 test − OD450 NC OD450 GC − OD450 NC) × 100, where OD600 and OD450 are optical density at 600 and 450 nm, respectively.To validate the performance of the newly developed resazurin-based microdilution assay, the susceptibility of 10 eviously . Assays As seen in M. mycetomatis isolate. The lowest MIC50s were obtained for the azoles, itraconazole, and posaconazole, followed by voriconazole (50s were obtained for amphotericin, terbinafine, and micafungin. The MICs reported here were comparable with those reported for itraconazole, posaconazole, and voriconazole (5–M. mycetomatis isolates by either the azole class of antifungal agents or terbinafine offers the best mechanism by which to inhibit M. mycetomatis growth (M. mycetomatis for any of the antifungal agents tested, the absolute value of the MIC is not indicative of therapeutic success or failure (–M. mycetomatis infection than previously reported (M. mycetomatis isolates tested were resistant to micafungin, in agreement with earlier studies (The MICs of the different antifungals used were variable and dependent on the conazole . Higher conazole , 5–10. As growth . Because failure –13. As rfailure –. Howeverfailure –. Based ofailure –. Amphotereported . It has reported . Apparen studies .The assay proved to be reproducible with high percent agreements . The perin vitro susceptibility assays for M. mycetomatis infection. Our in vitro susceptibility assay is flexible and can be used to test the antifungal activity of any compound and thus for drug discovery, making this assay more flexible than the commercially available Sensititre YeastOne assay (Our results indicate that resazurin is a good viability dye to be used in ne assay , which uin vitro susceptibility assays for M. mycetomatis based on resazurin as a viability dye offers an extra advantage of visual endpoint reading without resorting to spectrophotometric measurements. The aforementioned advantage coupled with the use of the less costly resazurin, as well as the flexibility with homemade plate layouts, would be an advanced step forward for in vitro susceptibility testing against M. mycetomatis compared with MTT, especially in endemic settings.In conclusion,


Gentiana macrophylla in Yulong County, China. The pathogenic nematodes were identified by observing morphological characteristics of females, second-stage juveniles and perineal pattern, sequence alignments, and specific amplification of sequence characterized amplified region (SCAR). The results showed that the perineal pattern of this nematode was round or oval, the dorsal arch was moderately high or low, one side or both of the lateral field extended to form a wing shape, the tail region had punctations, and the morphological characteristics and morphometric values of second-stage juveniles and females were similar to those of Meloidogyne hapla. The ITS region fragment of this nematode were highly similar to those of M. hapla in NCBI database, with a similarity of over 99.35%. Using the SCAR specific primers, a specific band with an expected size of approximately 440 bp was amplified from this nematode. Morphological and molecular identification supports the nematode species found on Gentiana macrophylla as M. hapla. This is the first report of this regulated root-knot nematode on Gentiana macrophylla in China.In September 2020, samples of galled roots with rhizosphere soil were collected from declining  Gentiana macrophylla) is a perennial herb of the Gentianaceae family, mainly produced in Mongolia, Russia, and China. Gentiana macrophylla has been widely cultivated in the southwest region of China for medicinal uses (Meloidogyne spp.). In order to clarify the nematode species of Gentiana macrophylla, extracted root-knot nematodes were identified by both molecular and morphological methods as Meloidogyne hapla.Qinjiao  were infested with 1,500 M. hapla J2 and maintained at 20 to 25°C in a greenhouse. The test was repeated three times with an inoculation with sterile water as a control.The nematodes were collected from the soil in the root zone using standard procedures . For morn = 20) was 53.5 ± 14.5 and egg masses 15.1 ± 3.20. The second-stage juveniles (J2) were collected from the soil in the root zone using standard procedures  were pear-shaped, 532.62 to 767.45 μm (659.03 ± 72.21) in length, and 243.64 to 557.52 μm (372.79 ± 67.42) in maximum body width, with round perineal pattern, low dorsal arch, and characteristic punctations present near the anus (n = 20) varied from 339.25 to 406.44 μm (372.32 ± 16.92), body width was 13.43 to 18.73 μm (14.48 ± 1.22), stylet length 10.21 to 14.45 μm (13.38 ± 0.87), tail length 36.66 to 51.43 μm (42.43 ± 4.58). These morphological characteristics are consistent with Meloidogyne hapla as described by M. hapla sequences. Additionally, a specific band with an expected size of approximately 440 bp was amplified by using the SCAR specific primers  and females.