Purpose: To determine and characterize the radiomics features from structural MRI (MPRAGE) and Diffusion Tensor Imaging (DTI) associated with the presence of mild traumatic brain injuries on student athletes with post-concussive syndrome (PCS).Material and Methods: 122 student athletes , median (IQR) age 18.8 (15–20) years, with a mixed level of play and sports activities, with a known history of concussion and clinical PCS, and 27 , median (IQR) age 20  years, concussion free athlete subjects were MRI imaged in a clinical MR machine. MPRAGE and DTI-FA and DTI-ADC images were used to extract radiomic features from white and gray matter regions within the entire brain (2 ROI) and the eight main lobes of the brain (16 ROI) for a total of 18 analyzed regions. Radiomic features were divided into five different data sets used to train and cross-validate five different filter-based Support Vector Machines. The top selected features of the top model were described. Furthermore, the test predictions of the top four models were ensembled into a single average prediction. The average prediction was evaluated for the association to the number of concussions and time from injury.Results: Ninety-one PCS subjects passed inclusion criteria . The average prediction of the top four models had a sensitivity of 0.80, 95% CI:  and specificity of 0.74 95%CI  for distinguishing subjects from controls. The white matter features were strongly associated with mTBI, while the whole-brain analysis of gray matter showed the worst association. The predictive index was significantly associated with the number of concussions (p < 0.0001) and associated with the time from injury (p < 0.01).Conclusion: MRI Radiomic features are associated with a history of mTBI and they were successfully used to build a predictive machine learning model for mTBI for subjects with PCS associated with a history of one or more concussions. Mild traumatic brain injuries (mTBI) or “Concussion” with Glascow Coma Scale scores of 13 or greater result in transient alterations in brain function with or without loss of consciousness . MoreoveRoutine clinical MRI studies do not show abnormalities in most SRC and PCS patients, however, diffusion-weighted (DWI) and diffusion tensor imaging (DTI) have shown variable localized, diffuse, and widespread changes after mild injury , 10. PriAt present, many research programs are trying to use AI including deep and machine learning, which have been applied to medical image data in other neurological diseases, to develop validated clinical image-based tools to identify post-concussive changes, to correlate them to clinical symptoms, and to predict recovery time and clinical outcomes , 13. SevHere we evaluated whether radiomics and ML can be used to differentiate patients that suffered mTBI with subsequent post-concussion syndrome from concussion-free athletes. To achieve the aim of this study, a small set of student athletes with SRC-related post-concussion syndrome were recruited, as well as a small set of concussion-free student athletes as controls. The data population was studied with state-of-the-art clinical MRI pulse sequences, quantitative image analysis, descriptive statistical procedures, and machine learning (ML) models that enabled the detection of PCS in affected subjects. Finally, we examined the longitudinal behavior of the predictive index and its association with the number of concussions.This retrospective analysis was approved by the Research Subjects Review Board of the University of Rochester. The study consisted of 122 subjects with a mixed level of play and sports activities. The concussed subjects were 65 males and 57 females, with a history of concussion who were High School or College Athletes with a history of one or more concussions and post-concussive symptoms who were diagnosed and evaluated by Sports Medicine Physicians; Physical Medicine, and Rehabilitation Physicians, or Neurologists. These patients were referred for MRI because of persistent clinical symptoms after mTBI from 2016 to 2019. The median (IQR) age for both male and female athletes was 17 15–20) years. All case subjects in our study were referred to the University outpatient imaging center and imaged on the same 3T MRI scanner due to incomplete resolution of symptoms after concussion. Inclusion criteria included a history of concussion and PCS, while exclusion criteria included dental braces, prior brain surgery, ventricular shunt, skull fractures, or standard contraindications for MR imaging. Diagnosis of concussion was made by ED physicians initially and PCS was determined by neurologists, physical medicine and rehabilitation physicians, and/or sports medicine physicians. The number of previous concussions, loss of consciousness (LOC) at the most recent concussion, and time between injury and MRI were extracted by a clinician from electronic medical records. All evaluations included the Acute Concussion Evaluation (ACE) , FOV = 250 mm, Flip angle = 8 degrees, 3D AXIAL SWI (88 slices/1.5 mm slice thickness (interleaved)/FOV 220 mm, TR 27 ms, TE 29 ms, 1 average), DOUBLE IR (FLAIR)-Fat-sat FLAIR 120 slices, 1.4 mm slice thickness, FOV 260 mm, TR 7,500 ms, TE 321 ms, TI 1: 3,000 ms; TI 2: 450 ms, 1 average, Acceleration factor 2, ref lines 24, Turbo Factor 256.2; b-value 2: 1,000 s/mm2 GRE Field Mapping for geometric and eddy current corrections 86 slices, FOV 256 mm, 2 mm slice thickness, TR 838 ms, TE1: 4.92 sec, TE2: 7.38 ms, Flip angle: 60°. All image sets used in this study were anonymized.DTI acquisition parameters were AXIAL DTI/ TA: 10:14 min,70 slices, FOV 256, 2 mm slice thickness, TR 9,000 ms, TE 88 ms, Flip Angle 15°, 1 average, Acceleration Factor 2/ref lines 24, Diffusion directions 64, b-value 1: 0 s/mmi = {1, 2, 3} decomposition levels. The LFD and the three |WD|i were fused into a single 5-Dimensional MRI set for MPRAGE, DTI-FA, and DTI-ADC, hence creating a 3D volume where each voxel was described by 15 signal/textural features; i.e., signal dimensions. Once preprocessed, we proceeded to do the image segmentation. The segmentation process consisted of using the ICBM 152 Linear atlas and main lobe labels as a reference  for any artifacts that might affect the quality of the study, as well as for the presence of recent or remote intracranial hemorrhage, signal abnormalities in the brain, hydrocephalus, congenital or developmental anomalies. Furthermore, each one of them automatically segmented brain VOIs, and tissues were visually inspected to discard subjects with segmentation errors. Subjects with unknown concussions or injury time were also removed from this study.The 10 image features extracted from the 18 ROI and 15 signal dimensions created a 2,700 (10 × 18 × 15) feature vector to be analyzed at the Machine Learning stage. A knowledge-based feature reduction strategy was used to decrease the dimensionality of the quantitative set. Studies have shown that mTBI mainly affects white matter tissue . FurtherwhereTo explore the origin of the signal associated with mTBI, we divided the entire feature set into five different feature sets. t-student test, and the Minimum Redundancy Maximum Relevance (mRMR) (Each one of the five feature sets was modeled by a support vector machine (SVM) using the radial basis kernel function , with dee (mRMR) . The trap-value < 0.01) between gender-age-height and each feature were mitigated by the residual method: fitting linear regression models of the reference group, and subtracting the estimated association to each feature . The test results of sets 2, 3, 4, and 5 were ensembled to get an estimation of a PCS index:Due to the associations of brain structures with development age, body height, and weight , we adju feature . Once asp(PCS) is the ensembled probability of a subject affected by PCS.where p-values of trends between concussion index and the number of concussions and days from injury. We explored gender differences by running the analysis on both males and females.The index was analyzed for associations to the number of subject concussions and the days from injury on subjects with <4 concussions. We used the spearman regression test to compute the p < 0.05). p < 0.001). The characteristics of the selected training cases are described in p = 0.002).Four subjects had to be removed for lack of complete clinical data. Twenty MRI data sets had severe artifacts due to the use of a defective head-neck coil where the neck receiver induced artifacts, and 11 subjects were older than 29 years, hence we removed them from the set. None of the subjects had signal abnormalities in the brain including sites of hemorrhage using the standard conventional (non-DTI) MR imaging sequences. p < 0.001). p < 0.009). p < 0.001), while males did not (p = 0.25). Regarding time from injury, the PCS gender stratified index was not statistically associated with days from injury for females (p = 0.22) nor males (p = 0.09).This proof of principle study showcased the potential of radiomics/ML in the understanding and diagnosis of PCS from MRI series, supporting the findings of DTI and DWI studies that indicate disruption of tract integrity and conductivity on PCS patients , 13, 29.We followed a radiomics approach similar to the efforts in developing predictive and diagnostic tools for brain cancers  and degeThe feature selection process identified top-performing radiomic features from the structural MPRAGE series as well as from DTI-FA and DTI-ADC images. This finding indicates that in addition to the detectable signal intensity and the texture changes in diffusivity maps, the signal intensity behavior has also changed in MPRAGE structural images, even if unable to be visually detected. That MPRAGE finding supported previous studies showcasing the potential of signal and textural information of structural imaging in the study of other brain conditions , 34.The 1,000 hold-out random test results showed consistent behavior of all the trained models. The white-matter model was superior to the gray-matter model, confirming previous findings indicating that white matter is the primary tissue affected by mTBI , 22, 29.Although this material does not include follow-up imaging studies, the findings showing that subjects who were imaged weeks, months, or even over a year post-injury had abnormal findings in FA, ADC, and structural images are consistent with findings from previous studies where follow-up analysis showed long-lasting FA and ADC changes in the brain , 11. TheThe reported findings should be contrasted with alternative PCS diagnostic tests. Premorbid health conditions have been associated with severity of symptoms –40 as weRegarding limitations, all subjects and controls were imaged with the same MRI scanner using the same imaging protocols which provided data harmonization but limits the assessments of clinical validity in a multicenter study. Furthermore, the study did not aim to address the hypothesis of imaging findings and their association to ongoing neural repair; hence we lack evidence that current imaging findings are associated with premorbid conditions or other neural factors. Furthermore, the reported performance is still far from being applicable in a clinical care setting that usually requires 90% accuracy; henceforth the provided method may be used to ease patient concerns and near future estimations of symptom resolution. This study has other limitations: First, the sample size is relatively small, and we did not have a large reference control group. Second, the age of the controls was slightly different from that of the study subjects; hence, the discovered PCS models may be still affected by the imperfect age match and not ready for any clinical use. Third, this study lacked longitudinal observations and was not designed to test longitudinal associations. Therefore, the observed time associations of the PCS index may be a modeling artifact, thus we can't reach strong conclusions regarding the potential of MRI signatures to monitor PCS resolution. Henceforth, these study results should be reproduced and validated in a large independent cohort with longitudinal and serial MRI observations of subjects with and without persistent symptoms. Future work must include direct comparisons to blood biomarkers and premorbid Behavioral Health conditions and their combination in a multimodal setting.This proof of principle study showed that MRI radiomic features from both diffusion and MPRAGE images were different in PCS subjects from trauma-free controls. Following an ML strategy, we modeled a classifier that was able to predict the presence of concussion on 81% of the concussed subjects with a specificity of 74%. The findings suggest that the concussion-induced abnormalities on PCS subjects are not uniformly distributed among the entire brain tissue. Furthermore, this study indicated that a signal build from these MR features accumulates with the history of previous concussions. This proof of principle but tightly controlled study has shown that PCS subjects may have localized brain abnormalities, which may be invisible to conventional radiologic observation, but are present and able to be detected with radiomic feature analysis.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 University of Rochester Office for Human Subject Protection. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.JT-P worked on the statistical analysis, drafting the first version of the paper, and reviewing the final manuscript. SM contributed to reviewing the paper and provided all the MRI images and the IRB. PR contributed to patient recruitment. ES managed the project and reviewed the manuscript. PG processed the images and reviewed the manuscript. AV designed the pulse sequence and reviewed the manuscript. ST processed the images and reviewed the quality of the MRI images as well as the segmentation. All authors contributed to the article and approved the submitted version.JT-P, ST, ES, and PG are shareholders of Qmetrics Technologies. The remaining 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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Rhodoluna (R.) lacicola strain MWH-Ta8 and Aurantimicrobium (A.) photophilum strain MWH-Mo1 grow faster in the light than in the dark, but do not use light energy to support growth. Here, we characterize transcription throughout a light-dark cycle in R. lacicola and A. photophilum. In both species, some genes encoding carbohydrate metabolism and storage are upregulated in the light. However, expression of genes of the TCA cycle is only coordinated with light availability in R. lacicola. In fact, the majority of genes that respond to light and darkness in these two species are different, even though their light-responsive phenotypes are similar. The ability to respond to light and darkness may be widespread in freshwater Actinobacteria, but the genetic networks controlled by these two stimuli may vary significantly.Light is a ubiquitous source of both energy and information in surface environments, and regulates gene expression not only in photosynthetic microorganisms, but in a broad range of photoheterotrophic and heterotrophic microbes as well. Actinobacteria are keystone species in surface freshwater environments, where the ability to sense light could allow them to coordinate periods of nutrient uptake and metabolic activity with primary production. The model freshwater Actinobacteria  Light is a ubiquitous resource in surface environments, and widely used by microbes. In fact, light-sensing proteins that control gene expression are common in photosynthetic microbes, photoheterotrophs, and non-phototrophic heterotrophs . The orgIn illuminated freshwater environments, Actinobacteria in the Microbacteriaceae family are ubiquitous and abundant keystone species which mediate fluxes of organic carbon and nitrogen, reduced sulfur, and vitamins . The freRhodoluna (R.) lacicola strain MWH-Ta8 and Aurantimicrobium (A.) photophilum strain MWH-Mo1, grow faster in blue light than in the dark, even though neither has a functional rhodopsin under laboratory conditions , and the two genomes share 879 genes, representing 65% of the R. lacicola genome and 50% of the A. photophilum genome  and placed on an orbital shaker in a dark room. White light was provided by four 13-watt compact fluorescent bulbs (each providing ∼30–40 μmol photons m–2 s–1) arranged approximately 30 cm above the cultures, on a 12-h ON/12-h OFF cycle. The cultures acclimated to the light/dark cycle for 24 h before sampling began. The first sample was taken in the dark, ∼5 min before the light turned on (t0). Then, samples were taken 15 min , 1 h (t2), and 6 h (t3) after the light turned on, and 15 min , 1 h , and 6 h  after the light turned off. For each time point and each strain, the entire culture (50 mL) was centrifuged at 4,500 × g for 30 min at room temperature. The supernatant was removed and the cell pellet was resuspended in 1 mL RNAlater (Invitrogen) and stored at −20oC until processing. Biological replicates were obtained by repeating the experiment 3 times for a total of 4 replicates, ensuring that the initial concentration of cells remained the same.Y medium  at room Y medium . For theR. lacicola sample and from two pooled aliquots of A. photophilum samples. The remainder of the material was archived at −80oC.The archived cell suspensions were separated into four aliquots, and RNA was extracted from one aliquot of each R. lacicola was extracted as described previously  with lysozyme (15 mg mL–1) and proteinase K (2 mg mL–1). Immediately prior to sonication, Buffer RLT (Qiagen) was added. Cells were then lysed using a Branson Sonifier 450 equipped with a microtip . After sonication, 500 μL 100% ethanol was added, and purification of total RNA continued using the RNeasy Mini Kit (Qiagen 74104) as described previously  and RNA quality was assessed using an AATI fragment analyzer. RNA was concentrated using the RNeasy MinElute Cleanup kit (Qiagen 74204) and quantified using the Qubit RNA BR assay kit (Invitrogen Q10210). The samples sent to the Joint Genome Institute (JGI) for library preparation and sequencing were 25 μL each, containing 25–359 ng RNA μLLibrary preparation and sequencing was carried out at the JGI. For each sample, rRNA was removed from 100 ng of total RNA using the Ribo-Zero Bacterial rRNA Removal Kit (Illumina). Stranded cDNA libraries were generated using the Illumina Truseq Stranded mRNA Library Prep Kit. The rRNA-depleted RNA was fragmented and reverse transcribed using random hexamers and SSII (Invitrogen) followed by second strand synthesis. The fragmented cDNA was treated with end-repair, A-tailing, adapter ligation, and 10 cycles of PCR. The prepared libraries were quantified using KAPA Biosystem’s next-generation sequencing library qPCR kit and run on a Roche LightCycler 480 real-time PCR instrument. Sequencing of the flowcell was performed on the Illumina HiSeq2500 sequencer using HiSeq TruSeq SBS sequencing kits, v4, following a 2 × 150 nt indexed run recipe.Standard pre-processing and filtering were done at the JGI using standard JGI pipelines. Raw fastq file reads were filtered and processed using BBDuk  and its A. photophilum samples  were excluded from downstream analyses because they did not correlate well with their replicate group, as determined by a Pearson correlation calculation.The raw reads from each library were aligned to the reference genomes using BBMap  with onlA. photophilum, this analysis showed that 4 samples from different treatment groups separated from all other samples. These four samples  were located in the same column of the sequencing plate, and data from these samples was also excluded from further analysis. With the 4 samples removed in the earlier stage, this resulted in exclusion of a total of 8 samples, leaving 20 samples from A. photophilum for downstream analysis.To evaluate the similarity of the replicate samples, the plotPCA function of the BiocGenerics R package version 0.34.0  was usedp-value ≤ 0.015.For differential expression analysis, raw gene counts were input to the DESeq2 R package, version 1.28.1 , which fTo group genes by expression patterns throughout the time series, rlog-transformed count values of differentially expressed genes were input to the degPatterns function of the DEGreport R package, version 1.24.1 . The deg–3. The GO-slim function was applied to generate general GO-slim terms to obtain an overview of the biological processes, cellular locations, and molecular functions of predicted proteins. Gene groups were merged based on their mean expression pattern.Gene Ontology (GO) annotations were assigned to identify functions in gene clusters according the GO database, which is the key functional classification of the National Center for Biotechnology Information (NCBI) . ProteinTo determine enrichment of a GO-slim term in a merged gene group, odds ratios were calculated as the ratios of representation of a GO term within a merged group to the representation of the same term in the genome  The stats (version 4.0.4) R package was used to calculate the odds ratios and do a one-sided Fisher’s exact test (using the fisher.test function) to test for significance. Results were then filtered with criteria odds ratio > 1, a count in the merged group > 1, and a count in the whole genome ≥ 10.Gene expression patterns were plotted as Z-score transformed transcript abundances. Z-score transformation standardizes data across sampling time points and allows for analysis that is independent of the raw count transcript abundance. Z-score were calculated by subtracting a gene’s mean transcript abundance across all timepoints from each sample, then dividing by the standard deviation of all timepoints. Z-scores for timepoint replicates were then averaged. The Z-score represents the number of standard deviations away from the mean transcript abundance across all timepoints. Rlog transformed transcript abundance values above the mean have a positive Z-score while rlog transformed transcript abundance values below the mean have a negative Z-score.All plots were generated using the ggplot2 R package, version 3.3.3  and the R. lacicola, we obtained a total of 718,789,878 raw reads  and 653,982,504 reads passed quality filtering , resulting in 326,991,252 total fragments   were sequenced by the JGI using an Illumina HiSeq system, paired-end 150-nt reads. For 650,221) . 326,304A. photophilum, we obtained 547,198,496 raw reads  and 455,067,112 reads  passed quality filtering, resulting in 227,533,556 total fragments  (For  sample) . A totalR. lacicola, PC1 best captures variance among light timepoints  groups closest to the first two light timepoints, possibly indicating a cyclical expression pattern. The replicates roughly group together and lie closest to their neighboring timepoints. In A. photophilum, PC1 captures the variance between light and dark timepoints  and grouped together along both PC1 and PC2, rather than with their timepoint replicates. Removing those samples left three replicates for each timepoint, except for t5, which had two replicates in downstream analyses. The two remaining t5 replicates cluster tightly together along PC1 and PC2 that collectively represent 76% of the variance. Of the top 100 genes with the highest loading values in PC1 and PC2, PC2 loading values were the most affected by removing the four samples. Nevertheless, PC2 captures variance between the early and late dark samples both before and after removing the outlying samples.Regularized logarithmic (rlog) transformed count data was analyzed via a principal component analysis to compare treatment groups. In mepoints . The firmepoints , indicatR. lacicola and 0.995 for A. photophilum, indicating very strong cluster structures in both species . Fourteen expression groups were identified in each strain , putative dark-responsive groups (2), and 4 groups which may respond to all changes in light availability . A second peroxiredoxin, Rhola_00011030, does not fall into an expression group, but its expression clearly follows a light-dark cycle  and putative components of a tad/flp pilus (two homologs of tadA and one tadBC homolog). Although not all of the genes putatively encoding components of the tad/flp pilus in R. lacicola fall into specific expression groups, expression of all except comEA increases sharply within 1 h of the light turning off , phrB (rhola_00007890), cryB (rhola_00013030), and cpf2 (rhola_00011000), encoding a putative DNA photolyase, cryptochrome, and cryptochrome-photolyase family protein, respectively . In R. lacicola, ribAB, ribD, and ribE fall into expression group 3J, and although ribH is not in that group, it shares the same expression pattern: transcript abundance increases between t0 and 15 min in the light, then decreases until 1 h in the dark, then increases again  increases early in the light period, then decreases until the light turns off, then increases again. In contrast, a cluster of three genes encoding a phytoene desaturase and two putative lycopene cyclases decreases in the light, increases in the dark, and a second cluster with a second putative phytoene destaturase and a prenyltransferase increases early in the light period, then decreases for the rest of the experiment, with a transient increase early in the dark period  and their genomes are broadly similar. We had also expected that light would stimulate or repress expression of specific genes, and that the effect would decay over time. Instead, we found that different pathways respond to light in the two strains, and that light and darkness alter transcription differently in both strains.We hypothesized that 1F0 ATP synthase. In R. lacicola, expression of the genes of the TCA cycle and the ATP synthase clearly increases in the light and decreases in response to darkness. Further, the expression patterns of carbohydrate metabolism genes suggest that R. lacicola may prioritize early steps of carbohydrate metabolism (glycolysis) in the dark and the TCA cycle as well as synthesis of carbohydrates and storage molecules such as starch in the light. In A. photophilum, there are three different expression patterns among the TCA cycle genes, but only one  was included in an expression group  in A. photophilum has a similar expression pattern to the carotenoid biosynthetic genes. In R. lacicola, actR expression is similar to expression of about half of the carotenoid biosynthetic genes; the different expression patterns observed in this pathway suggests that different carotenoid products may be produced at different times.Some interesting similarities between the two strains occur in expression of potential light-sensitive proteins and the genes with similar patterns. Expression of the rhodopsin in both strains increases greatly in the dark, and the heliorhodopsin  that control circadian rhythms in cyanobacteria, and that the only predicted proteins with homology to light-sensing domains such as PAS, GAF, and BLUF domains are the photolyases and cryptochromes .The datasets presented in this study can be found in online at the NCBI sequencing read archive under the Bioproject Numbers JM designed the experiments, supervised the laboratory work and data analysis, and led the writing effort. JK did the laboratory experiments and contributed to writing and editing. PH did the bioinformatic analysis and contributed to writing. All authors contributed to the article and approved the submitted version.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


We present a case of hemichorea in a patient with nonketotic hyperglycemia (NKH), a rare presentation of hyperglycemia. A 55-year-old female with diabetes presented to the emergency department with involuntary bilateral upper and lower limb movements for five days. The patient had a serum glucose level of 358 mg/dL (19.87 mmol/L) and improved after controlling blood sugar levels. When we encounter a case of chorea in the emergency department, high blood sugar levels are an essential underlying reversible etiology to be kept in mind. India has a prevalence of diabetes of 8.7 . Type 2 Chorea is a hyperkinetic movement disorder characterized by rapid nonpurposive movements of distal limbs and can involve the face and trunk. It can be caused by neurodegenerative, cerebrovascular, immunological, neoplastic, infectious, and metabolic diseases. Chorea following NKH has favorable outcomes after correction of hyperglycemia. Here, we present a case of an elderly female from a hilly region of Uttarakhand, manifesting as NKH chorea. Clinicians should be aware of this rare entity for early treatment.A 55-year-old female patient presented to the emergency department with complaints of acute onset generalized involuntary choreiform movements for the last five days. It started as bruxism and progressed to involve bilateral upper limbs and left lower limb and was associated with difficulty in speaking. It was noted that the movements resolved while the patient was sleeping. It was not associated with loss of consciousness, bladder or bowel incontinence, or weakness of any body part.The patient had no history of fever, headache, vomiting, seizures, rash, drug intake, falls, and head injury and has been diagnosed with diabetes for 14 years on irregular treatment with insulin. There was no history of similar illnesses in the family. On examination, the patient was conscious, oriented, with a Glasgow Coma Scale (GCS) score of E4V5M6, blood pressure of 110/70 mmHg, respiratory rate of 20 cycles/min, SpO2 of 96% on room air, and pulse rate of 81 beats/min. The patient's bilateral pupils were reactive and of normal size and the patient had normal vesicular breath sounds. The abdomen was soft and nontender. Cardiovascular system examination revealed normal heart sounds. Regarding the central nervous system, choreiform movements in bilateral upper limbs and left lower limb were observed, and the power in all four limbs was 5/5. The tone was normal, and the bilateral plantar were flexor. Cranial nerve examination was normal. Cerebellar signs were absent. Video Investigations revealed mild anemia, and other blood tests were within normal limits Table .Non-contrast computed tomography of the head revealed a tiny calcified granuloma in the right parietal lobe. Magnetic resonance imaging of the brain suggested a hyperintense lesion in the bilateral basal ganglia on T1, indicating a metabolic insult. Figure Random blood sugar level was 358 mg/dL (19.87 mmol/L) with negative urine ketones and normal arterial blood gas analysis . The patient's glycated hemoglobin (HbA1c) was 10.4%; thus, we made the diagnosis of NKH chorea. In the emergency department, isotonic saline (0.9% NaCl) at the rate of 10-20 ml/kg and insulin was administered to control blood sugar levels; and haloperidol (5 mg) was intravenously injected to control the involuntary movements of the patient. The patient was discharged with clonazepam (10 mg per tablet) along with insulin (subcutaneous). The patient is now asymptomatic with regular follow-up in the neurology outpatient department.Chorea, athetosis, and ballism are the involuntary movements of the choreiform spectrum. NKH chorea encompasses a triad of chorea, hyperglycemia, and basal ganglia hyperintensity and is a rare complication of diabetes mellitus. The prevalence of NKH chorea is 1/100,000 and Bedwell, in 1960, has first reported NKH chorea as a rare clinical entity ,3. Oh etThe underlying pathogenesis of chorea in NKH chorea is attributable to hyperglycemia-induced basal ganglia dysfunction. Hyperglycemia leads to the stimulation of the anaerobic pathway in the brain, leading to Krebs cycle inhibition. The metabolic demand of the brain is fulfilled by converting gamma aminobutyric acid (GABA) to succinic acid, resulting in metabolic acidosis. In ketotic hyperglycemia, GABA is resynthesized; however, in NKH, both GABA and acetate are severely depleted, resulting in basal ganglia dysfunction . Chorea One radiological feature of NKH chorea is basal ganglia hyperintensity on T1-weighted MRI. A similar finding was observed in patients with hepatic encephalopathy, hypoglycemic coma, and post-cardiac arrest encephalopathy . Most paNKH chorea is a rare complication of uncontrolled diabetes mellitus and is more commonly seen in elderly females. It is one of the reversible metabolic causes of chorea; timely recognition and appropriate treatment of this condition will improve the patient's quality of life.


Diabetes has become a global public health problem and danger to human health. Diabetes is the main cause of blindness, kidney failure, heart attack, stroke, and lower limb amputation. According to the latest epidemiological survey and research, the overall prevalence of diabetes in mainland China is 11.2%, of which type 2 diabetes mellitus (T2DM) accounts for more than 90% acupuncture combined with Chinese herbal medicine have been widely used in the treatment of T2DM. However, we have not found a meta-analysis of their synergistic effects. Therefore, this systematic review and meta-analysis aims to evaluate the efficacy and safety of acupuncture combined with Chinese herbal medicine in the treatment of T2DM.From inception up to September 20, 2021, the PubMed, Web of Science, Embase, AMED, Cochrane Library, CNKI, VIP, CBM, and Wanfang databases will be searched. The publication date or language will not be limited. We will apply a combination of medical keywords, including “acupuncture”, “Chinese herbal medicine”, and “type 2 diabetes mellitus”. We will also check other ongoing and unpublished studies in the clinical trial registry. At the same time, we will manually search all reference lists from relevant systematic reviews to find other eligible studies. We will use Review Manager software (REVMAN v5.3 Cochrane Collaboration) to meta-analyze the selected literature. The study for acupuncture combined with Chinese herbal medicine in the treatment of T2DM was a randomized controlled study. Two researchers will independently review the research selection, data extraction, and research quality assessments. Finally, we will observe the outcome measures.This study will generate evidence-based data on the treatment of T2DM with acupuncture combined with Chinese herbal medicine and will provide new ideas and treatment modalities to investigate in future research. Although there are a wealth of drugs and methods for the treatment of diabetes, there are various insulin and oral hypoglycemic agents, such as biguanides, thiazolidinediones, α-glycosidase inhibitors, sodium-glucose cotransporter 2 inhibitors, sulfonylureas, glinide, and dipeptidyl peptidase IV inhibitors, that can effectively lower blood sugar in a short time, eliminate clinical symptoms, and delay the occurrence of complications. However, long-term use of these drugs also leads to drug resistance, and the incidence of diabetes and its complications is still on the rise. Obviously, diabetes has become a global public health problem and danger to human health.According to the World Health Organization, the prevalence of diabetes is increasing year by year, and its prevalence is rising faster in low- and middle-income countries than in high-income countries. Diabetes is the main cause of blindness, kidney failure, heart attack, stroke, and lower limb amputation. In 2019, it was estimated that diabetes directly caused 1.5 million deaths.,5 As a complementary and alternative medicine practice, Chinese herbal medicine has significant advantages in the treatment of diabetes and its complications.,7 However, there is no robust evidence showing the effectiveness of acupuncture combined with Chinese herbal medicine in treating T2DM and its side effects; this lack of evidence is also an important factor hindering its promotion in the treatment of T2DM and its complications. Therefore, this systematic review and meta-analysis will evaluate the efficacy and safety of acupuncture combined with Chinese herbal medicine in the treatment of T2DM.In recent years, acupuncture combined with Chinese herbal medicine have been widely used in the treatment of T2DM. As an important part of traditional Chinese medicine, acupuncture can correct various metabolic disorders such as hyperglycemia, overweight, excessive appetite, and hyperlipidemia. Acupuncture can increase insulin sensitivity and improve insulin resistance.22.1 This study is a retrospective study and meta-analysis, so the study design, process, and results do not require patient and public participation or ethical approval. From inception up to September 20, 2021, the PubMed, Web of Science, Embase, AMED, Cochrane Library, CNKI, VIP, CBM, and Wanfang databases will be searched. The publication date or language will not be limited. We will apply a combination of medical keywords, including “acupuncture”, “Chinese herbal medicine”, and “type 2 diabetes mellitus”. We will also check other ongoing and unpublished studies in the clinical trial registry. At the same time, we will manually search all reference lists from relevant systematic reviews to find other eligible studies. The expected registration has been approved by the International Platform of Registered Systematic Review and Meta-analysis Protocols. (https://inplasy.com/inplasy-2021-10-0015/). And the registration number is INPLASY2021100015. The search strategy for the PubMed is presented in Table This study will be based on the reporting guidelines for systematic review protocols and meta-analyses (PRISMA-P).2.2The inclusion criteria for the literature search will be as follows: studies including patients aged ≥18 years, regardless of sex, with T2DM diagnosed according to the standards of the American Diabetes Association or the World Health Organization; randomized controlled studies; studies in which the experimental group was treated with acupuncture combined with Chinese herbal medicine; studies in which the control group was treated with placebo or hypoglycemic drugs. The exclusion criteria will be as follows: studies including patients aged <18, with non-T2DM; studies in which the experimental group was given treatment methods other than acupuncture combined with Chinese herbal medicine; studies in which the control group was not treated with a placebo or a hypoglycemic drug; studies that were not randomized controlled trials.2.3 If the full text or the required information on the analytic process is missing from a paper, the author of the study will be contacted, and the data will be requested. The 2 researchers will independently evaluate the methodological quality of the included studies according to the Cochrane manual guidelines, and they will report the results according to the PRISMA guidelines. During the research period, any disagreements between the researchers will be resolved through discussion or negotiation with a third researcher until a consensus is reached. A flowchart of the screening process is presented in Figure The 2 researchers will independently carry out the literature search; screening and data extraction will be based on pre-established search strategies, literature inclusion and exclusion criteria, and data extraction tables. We will exclude all conference records, reviews, meta-analyses, newspapers, guides, letters, and other documents. The research selection process will be represented by the PRISMA flowchart.2.4 The assessment content addresses the following: whether the random method is correct; whether the allocation concealment is achieved; whether the blind method is implemented; whether the result data are complete; whether there is selective reporting bias; and whether there are other biases. We will use Begg and Egger tests and set P < .1 as statistically significant, and we will use a funnel chart to assess publication bias. If the evaluation quality of any single study is inconsistent between researchers, the discrepancy will be resolved through consensus between the researchers.The 2 researchers will use the Cochrane risk bias assessment tool to separately assess the quality of the randomized studies.2.5The primary outcome measures will include fasting blood glucose, 2-hour postprandial blood glucose, and glycosylated hemoglobin. The secondary outcome measures will include triglycerides, cholesterol, and insulin secretion. Adverse events resulting from safety violations will be reported.2.6P < .05 will be considered statistically significant. The 2 researchers will independently carry out data extraction and input, and a third researcher will conduct inspections, while the first 2 researchers will conduct data calculations. Quantitative data express the combined effect size with standardized mean square error and 95% confidence interval. Binary categorical variables express the combined effect size by odds ratio and 95% confidence interval. The heterogeneity among the included research results will be analyzed by the I2 test. If P > .05 and I2 < 50%, there is homogeneity among the studies, and the fixed-effects model will be used for meta-analysis. If P ≤ .05 and I2 ≥ 50%, there is heterogeneity among the studies. Sensitivity analysis will then be used to analyze the source of heterogeneity. After the influence of clinical heterogeneity is excluded, the random-effects model will be used for meta-analysis. We will use subgroup analysis based on different interventions, controls, and outcomes.We will use Review Manager software (REVMAN v5.3 Cochrane Collaboration) to meta-analyze the selected literature, and 3,13 Although many new drugs for the treatment of T2DM, such as glucagon-like peptide 1 receptor agonists and sodium-glucose cotransporter 2 inhibitors, have been successfully developed, researchers are looking for better blood-glucose-lowering treatment programs. Controlling glucose levels and reducing complications from T2DM involves continual disease management. In the treatment of T2DM, acupuncture combined with Chinese herbal medicine can reduce blood sugar, eliminate clinical symptoms, increase insulin sensitivity, and improve insulin resistance. They can also alleviate the side effects caused by hypoglycemic drugs. Therefore, acupuncture combined with Chinese herbal medicine has broad potential applications in the treatment of T2DM. This study will generate evidence-based data on the treatment of T2DM with acupuncture combined with Chinese herbal medicine and will provide new ideas and treatment modalities to investigate in future research.Diabetes is a global public health problem that affects human physical and mental health, and it is also an independent risk factor for major diseases such as kidney failure, cardiovascular disease, and stroke.All the authors of this manuscript are very grateful to the various departments of Changchun University of Chinese Medicine for their support.Conceptualization: Pengjie Bao, Zheng Nan.Data curation: Pengjie Bao, Jia Mi, Ziyang Yu.Formal analysis: Le Liu, Zhiyue Zhu.Funding acquisition: Jia Mi.Investigation: Pengjie Bao, Shilin Liu.Methodology: Pengjie Bao, Zheng Nan.Project administration: Jia Mi, Zheng Nan.Supervision: Jia Mi, Zheng Nan.Validation: Le Liu, Zhiyue Zhu, Shilin Liu.Visualization: Pengjie Bao, Jia Mi, Ziyang Yu.Writing – original draft: All authors.Final approval of manuscript: All authors.


Ascomycota, while the HS samples were predominated by Mortierellomycota and Basidiomycota. The major species in the LS samples were Plectosphaerellacucumerina and Trichocladiumasperum, whereas the dominant species in the HS samples were Epicoccum nigrum and Cladosporium chasmanthicola. These results suggested that the LS samples and the HS samples had different rhizosphere soil fungal community and composition changes resulting from implementation of FLC in potato growing areas.The application of herbicides to arable land is still the most effective and accepted method to protect plants from weeds. Extensive use of chemicals in conventional agricultural practices has resulted in continuous and serious environmental pollution. Flurochloridone (FLC) is a monophenyl pyrrolidinone selective herbicide that is commonly used to inhibit weeds that occur during the growth of potatoes. In recent years, research on the toxicity of FLC has gradually increased. However, it is relatively rare to analyze the role of FLC by studying the composition of soil microorganisms. Therefore, we used NGS methods to identify the fungal community structure of the low content soil (LS) and high content soil (HS) samples in this study. Subsequently, we identified the fungal community and composition differences of these two group samples using the statistical analysis. Despite the variances of fungal community and composition across the different samples within the group, the fungal composition of the LS samples and the HS samples. LS samples were predominated by  Environmental stresses pose a serious threat to agricultural production globally, and herbicides are among the most widely applied agrochemicals, significantly increasing agricultural productivity and crop yields ,2,3,4. FThe application of herbicides to arable land is still the most effective and accepted method to protect plants from weeds . ExtensiThe rhizosphere zone, a microclimate of soil that surrounds and is influenced by plant roots, harbors a vast number of soil biota involved in complex biological and ecological processes, and it is considered one of the most dynamic interfaces among ecosystems ,15. The Recent methodological advances have enabled researchers to chart the full extent of soil microbial diversity and to build a more comprehensive understanding of specific microbial controls on soil processes. In particular, the next-generation sequencing (NGS) technique has been successfully applied the rhizosphere soil microbial community and composition, because it can provide more detailed information and in-depth microbial community insights when compared to other molecular biology methods ,24,25,26In this study, the NGS technique was used to analyze the role of FLC by studying the composition of rhizosphere soil fungal microorganisms in potato growing areas of Qinghai-Tibet Plateau in China. We systematically compared the differences of the fungal communities in FLC application soil. The fungal microbial compositions were determined by the NGS technique of the internal transcribed spacer (ITS) region. This work is to determine rhizosphere soil fungal community and composition changes resulting from implementation of FLC in potato growing areas. It aims to clarify the issues of its dosage, period of use, method of use and safety, and provide a data basis for the large-scale application of the agent in the Qinghai-Tibet Plateau and even the other potato fields.2 FLC. After 1, 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, and 48 days, each sample comprised three biological replicates, and the rhizosphere soil samples were collected, respectively. The roots of each plant were gently shaken to remove the redundant soil, and the remaining soil adhered to the root hairs was collected with a brush and sieved with a 2-mm aperture sieve. The specific method details refer to previous research [The soil sampling was conducted in a potato growing areas in Qinghai-Tibet Plateau, Qinghai province. The location was at a longitude of 89°35′–103°04′ E and latitude of 31°39′–39°19′ N. The FLC and other chemicals used in the present study were of analytical grade from Sinopharm Chemical Reagent Co., Ltd. . The experimental soil used in the study was castanozems. The soil was mixed thoroughly with 750  and 1125  g/hmresearch . In tota−1 HCl and 0.03 mol × L−1 NH4F and measured by a spectrophotometer. Soil TP was determined colorimetrically using the molybdate method. Total nitrogen (TN) content determined by Kjeldahl digestion method.In order to investigate the changes of physicochemical properties, seven soil physicochemical were analyzed. The soil pH was determined using a calibrated pH meter  by the microelectrode method . The soiTotal genome DNA from rhizosphere soil samples was extracted using the soil DNA extraction kit following the protocol provided by the manufacturer . DNA concentration and purity was monitored on 1% agarose gels. According to the concentration, DNA was diluted to 1 ng/µL using sterile water.® High-Fidelity PCR Master Mix ; 2 µM of forward and reverse primers, and about 10 ng template DNA. Thermal cycling consisted of initial denaturation at 98 °C for 1 min, followed by 30 cycles of denaturation at 98 °C for 10 s, annealing at 50 °C for 30 s, and elongation at 72 °C for 30 s. Finally, 72 °C for 5 min. The “barcode” is an eight-base sequence unique to each sample. Mix same volume of 1x loading buffer (contained SYB green) with PCR products and operate electrophoresis on 2% agarose gel for detection. PCR products was mixed in equidensity ratios. Then, mixture PCR products was purified with Qiagen Gel Extraction Kit .The internal transcribed spacer regions of the ribosomal RNA gene were amplified by PCR using the primers ITS1-1F-F CTTGGTCATTTAGAGGAAGTAA, ITS1-1F-R GCTGCGTTCTTCATCGATGC ,32. All ® DNA PCR-Free Sample Preparation Kit  following the manufacturer’s recommendations, and index codes were added. The library quality was assessed on the Qubit@ 2.0 Fluorometer  and Agilent Bioanalyzer 2100 system. Finally, the library was sequenced on an Illumina NovaSeq platform and 250 bp paired-end reads were generated.The sequencing libraries were generated using TruSeqhttp://ccb.jhu.edu/software/FLASH/, accessed on 22 May 2021) [http://qiime.org/scripts/split_libraries_fastq.html, accessed on 22 May 2021) quality control process [http://www.drive5.com/usearch/manual/uchime_algo.html, accessed on 22 May 2021) to detect chimera sequences, and then the chimera sequences were removed. Then the effective tags were finally obtained.The paired-end reads were assigned to samples based on their unique barcodes and truncated by cutting off the barcode and primer sequence. Paired-end reads were merged using FLASH  , a very  process ,34. In bhttp://drive5.com/uparse/, accessed on 22 May 2021) [The sequence analyses were performed by Uparse software  . Sequencay 2021) . Alpha dAs shown in An average of 89,010 per samples were obtained from the 108 samples  .These saSequencing the ITS gene of all the soil samples showed that the soil fungal communities in all samples covered 14 phyla, 42 classes, 99 families, 204 families, 394 genera, and 405 species.Ascomycota was the most important phylum in LS and HS samples. The relative abundance of Ascomycota in the LS samples was higher than that in the HS samples. In contrast, the relative abundance of Mortierellomycota and Basidiomycota in the LS samples was lower than that in the HS samples.At the phylum level A, AscomyAscomycota plays an important role in the degradation of rhizosphere soil organic matter. It is speculated that the decrease in relative abundance of Ascomycota may be related to the decline of soil organic matter and soil fertility [ertility ,43.Thelebolaceae, Nectriaceae, Mortierellaceae, Glomerellaceae, Plectosphaerellaceae, Chaetomiaceae, Didymellaceae, Hypocreales_fam_Incertae_sedis, Pyronemataceae, and Pleosporaceae). Overall, Thelebolaceae was the most abundant family in both LS and HS samples. The proportions of Thelebolaceae in the LS samples were higher than that in the HS samples.At the family level B, the moThelebolus was the most abundant in all the LS samples and the HS samples. The other genus with high abundance, included Colletotrichum, Fusarium, Plectosphaerella, Trichocladium, Acremonium, Pseudombrophila, Boeremia, Neonectria, and Cladosporium, respectively. In general, Thelebolus was more abundant in the LS samples than that in the HS samples, whereas Colletotrichum and Fusarium in the LS samples was less than in HS samples. At the genus level, Cadophora and Tetracladium were the dominant microorganisms among the plant mycobiota. Although their OTU abundances were different, both Cadophora and Tetracladium belong to Ascomycota, which is considered the most likely endophytic taxon to colonize plants [Ascomycota is widespread in soil and plants, such as forage, flowers, and crops [At the genera level C, Thelebe plants . Ascomycnd crops ,46,47.Thelebolus microsporus, Plectosphaerella cucumerina, Trichocladium asperum, Pseudombrophila hepatica, Boeremia exigua, Epicoccum_nigrum, Cladosporium chasmanthicola, Alternaria alternata, Metarhizium robertsii and Gibberella pulicaris  samples, while the Rozellomycotina_cls_Incertae_sedis was higher in the HS (H27) samples.The Linear Discriminant Analysis Effect Size (LEfSe) analysis was used to determine the significant difference of the fungal communities between the LS (L27) samples and the HS (H27) samples A,B. The The rhizosphere soil fungal community and composition take on the role of soil decomposers. Different types and numbers of fungal communities play an important role in the process of material and energy cycles. They can convert complex and refractory organic matter, which is transformed into nutrient elements that plants can use ,49,50.Ascomycota, while the HS samples were predominated by Mortierellomycota and Basidiomycota. The major species in the LS samples were Plectosphaerella cucumerina and Trichocladium asperum, whereas the dominant species in the HS samples were Epicoccum nigrum and Cladosporium chasmanthicola. These results provided a basic understanding of the fungal community and composition and the significant difference of fungal profiles in the LS samples and the HS samplesafter FLC application. However, the roles of these fungal communities in the rhizosphere soil in potato growing areasof the Qinghai-Tibet Plateau are still unknown, further research is needed to investigate the roles of these species in the rhizosphere soil.We used the NGS technique to identify the rhizosphere soil fungal community and composition in the LS samples and the HS samples, and compared their fungal microbial and physicochemical properties. The 14 phyla, 42 classes, 99 orders, 204 families, 394 genera, and 405 species were identified in all of the rhizosphere soil samples. Despite the variances of soil fungal communities and compositions across the different samples, the fungal community, and composition of the LS samples and the HS samples, the LS samples were predominated by


Solute carriers (SLCs) are important transmembrane transporters with members organized into 65 families. They play crucial roles in transporting many important molecules, such as ions and some metabolites, across the membrane, maintaining cellular homeostasis. SLCs also play important roles in hearing. It has been found that mutations in some SLC members are associated with hearing loss. In this review, we summarize SLC family genes related with hearing dysfunction to reveal the vital roles of these transporters in auditory function. This summary could help us understand the auditory physiology and the mechanisms of hearing loss and further guide future studies of deafness gene identification. Most of the SLC proteins have 12 transmembrane domains , with more than 400 members . Among these genes, five of them belong to the SLC family. Besides, another 13 SLC members were reported to be associated with hearing loss or involved in auditory organ development. These SLC genes, although most of them act as transporters, have different expression patterns in auditory organs and distinct function in hearing.Hearing is one of the most important sensory functions, and hearing loss would cause great inconvenience to the daily life of deaf people. The causes of hearing loss vary, from congenital to acquired impairments, but the defects of genes account for the majority. More than 200 hearing loss genes, including the syndromic and the nonsyndromic hearing loss genes, were identified in the last decades  gene, encoding the Na–K–2Cl cotransporter-1 (NKCC1), is mainly expressed in the stria marginal cell of the cochlea  gene, which encodes electroneutral cation-coupled chloride cotransporters, is very important in some physiological processes, such as cell volume regulation, modulation of intraneuronal chloride concentration, transepithelial ion movement, and blood pressure regulation . There a hearing . The sol cochlea , which i cochlea . In mice cochlea , and knobehavior . Besidesn humans . Therefon humans , which pslc17a8 gene, is exclusively expressed in hair cells and localized to the basal end of hair cells , encoded by the ir cells , and theebrafish . Similar stimuli , and alsn humans . In addiing loss . A recening loss .SLC22A4, also named as OCTN1, a 551-amino acid–long protein, is a pH-dependent organic cation transporter  and inflammatory cytokines, such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α)  . It was  disease . Mice wion model . All of SLC22A4 was found to be associated with hereditary hearing loss in humans. In mammalian auditory organs, SLC22A4 is expressed in all of the inner ear epithelia at the early stage, including the hair cells and spiral ganglion neurons; however, the expression is restricted at the apical surface of stria vascularis (SV) endothelial cells in the mature cochlea  gene, encoding the protein pendrin, is the causal gene of Pendred syndrome, which is a recessively inherited disorder with hearing loss as the obvious feature  (SLC26A4 mutation are likely to be resistant to high blood pressure (The  feature . In addie (EVAS) . It was pressure .SLC26A4 gene was found to be expressed in the thyroid at high levels, and it can help the thyroid follicular cells transport iodine  gene, is the most well-studied solute carrier in hearing-related research. In the mammalian inner ear, it is specifically expressed in the basolateral membrane of outer hair cells  protein, encoded by the ir cells , and defing loss . In nonmy organs . The proy organs . Unlike y organs , which wy organs . Indeed,y organs . Nonmammy organs  and a wey organs . Prestiny organs  and intey organs . Moreovey organs . Recently organs , providiExcept for the SLC members discussed above, which have been identified as hearing loss genes, some other SLCs, waiting to be verified as deafness genes, were reported to function in hearing-related processes.SLC4 family members are bicarbonate transporters and play vital roles in acid–base homeostasis . Among tSLC4A2 gene, encoding HCO3−/Cl− anion exchangers, was reported to be expressed in mammalian inner ear cells, including hair cells and supporting cells , which is important in maintaining intracellular pH homeostasis by exchanging one intracellular H+ for one extracellular Na+ genes was found in a boy with nonsyndromic hearing loss; however, no hearing impairment occurred in his brother, father, and grandfather who have the same translocation , without hearing deficits  and SLC16A10 , which share similarities with each other, prefer to transport the iodothyronines (T4 and T3)  . Distinc and T3) . As know and T3) . Unsurpr tissues , and the tissues .SLC19A1 transports folates but not thiamine, and the other two transport thiamine but not folates , were reported to be associated with thiamine-responsive megaloblastic anemia (TRMA) , of which  folates . Mutatioa (TRMA) , which iSLC26A4 and SLC26A5, discussed above, another gene, SLC26A2, encoding for a sulfate/chloride transporter, may be associated with hearing loss. In zebrafish, the slc26a2 gene was proven to be critical for otic development and hair cell survival, and loss of function of slc26a2 led to defective auditory organ development and impaired hearing  are included in this family . In addi hearing .The SLC44 family, with five members (SLC44A1–5), is a choline-like transporter family .SLC44A2 and blocking its transporter function , is a transmembrane glycoprotein. It was identified as the target of the Kresge Hearing Research Institute-3 (KHRI-3) antibody, which can lead to autoimmune hearing loss by binding to function . Moreoveing loss .SLC44A4 encodes the choline transport protein CTL4. Mutations in this gene were found in a Chinese family with postlingual nonsyndromic mid-frequency sensorineural hearing loss, and knockdown of the slc44a4 gene in zebrafish led to significant defects in the otic vesicle and lateral line neuromast development, accompanied by defective hearing , was found in the BVVL syndrome  syndrome is a rare neurodegenerative disease characterized by sensorineural hearing loss and a variety of cranial nerve palsies . Mutatiosyndrome , and a hsyndrome , indicatAccording to the World Health Organization (WHO), one in four, about 2.5 billion, people worldwide will be living with some degree of hearing loss by 2050, and nearly 60% of hearing loss is caused by genetic factors. Therefore, hereditary hearing loss is a serious problem that needs attention. In the last decades, more than 200 human hearing loss genes have been identified by scientists all over the world.The SLC family genes, with more than 400 members, encode different types of transporters that function in solute transporting. Deficiency of some of these genes would cause human diseases. Until now, 18 members, as listed in this review, in the SLC family were reported to be associated with the occurrence of hereditary deafness, including five known hearing loss genes, which indicated the crucial roles of solute carriers in hearing function. These SLCs have different expression patterns in the inner ear and have distinct function, and each of them is essential for normal hearing. Mutations in any of these genes would result in inner ear cell dysfunction or even cell death, leading to hearing impairment or hearing loss.In this review, we systematically summarized all of the SLCs involved in hearing, from their expression to function to the underlying mechanisms. This study would be helpful in understanding the roles of solute transporting for normal hearing, and it can also provide ideas for deafness gene identification.


Insulin-like growth factors (IGFs) and insulin-like growth-factor-binding proteins (IGFBPs) regulate cell proliferation and differentiation and may be of importance in obesity development. The aim of the study was to analyze the expression of chosen IGF-axis genes and the concentration of their protein products in 28 obese children (OB) and 34 healthy control (HC), and their correlation with essential parameters associated with childhood obesity. The gene expression of IGFBP7 was higher, and the expression of IGF2 and IGFBP1 genes was lower in the OB. The expression of IGFBP6 tended to be lower in OB. IGFBP4 concentration was significantly higher, and IGFBP3 tended to be higher in the OB compared to the HC, while IGFBP1, IGFBP2, and IGFBP6 were significantly lower, and IGFBP7 tended to be lower in OB. We found numerous correlations between IGFs and IGFBP concentration and obesity metabolic parameters. IGFBP6 correlated positively with apelin, cholecystokinin, glucagone-like peptide-1, and leptin receptor. These peptides were also significantly lower in obese children in our study. The biological role of decreased levels of IGFBP6 in obese children needs further investigation. Obesity is a complex condition with a serious impact on overall health, both physical and psychological. It is defined by the World Health Organization (WHO) as “abnormal or excessive fat accumulation that may impair health” . It is cUntil now, the role of the insulin-like growth factors’ axis and the individual markers and hormones associated with obesity in children are poorly studied ,11,12. IBecause the IGF protein family plays a key role in metabolic processes, we decided to examine the differences in the concentration of numerous components of the IGF axis  in obese children in comparison to healthy control and to analyze the correlations of these components with parameters, such as blood pressure, concentration of insulin, adipokines , and peptides regulating gastrointestinal tract . The peptides concentration was assessed both fasting and after an oral glucose tolerance test. Additionally, taking into account the importance of epigenetic factors, the expression of genes regulating studied peptides of the IGF axis were analyzed.Two study groups were recruited. The obesity group (OB) included 28 children, 12 boys and 16 girls, 4–17.8 (average 13.7) years old. All were patients of the Pediatric and Adolescents Endocrinology Department, Jagiellonian University, Medical College in Krakow. Obesity was defined as a BMI at or above the 95th percentile for children of the same age and sex. BMI was calculated by dividing a children’s weight in kilograms by the square of height in meters. The inclusion criteria were obesity (BMI Z-score > 2.0) developed before the age of puberty and negative medical history or any signs or symptoms of acute or chronic diseases, no drugs and dietary supplements, and normal diet. The exclusion criteria were obesity secondary to medical conditions , chronic systemic diseases, or drugs .The control group consisted of 34 healthy peers: 13 boys and 21 girls, aged 4.3–16.9 (average 11.8) years. The children were recruited from the families of the patients and children of medical staff, analogous to the study group in terms of age and sex, all having a negative medical history and without any signs or symptoms of acute or chronic diseases, including obesity.Height and body weight measurements were performed by an anthropometrist. Weight was measured to the nearest 0.1 kg, and height was measured to the nearest 0.1 cm using a stadiometer and a balanced scale. The body mass index (BMI) and BMI percentile/SDS were calculated using online WHO BMI calculators based on CDC growth charts for children and teens ages 2 through 19 years. The results were compared to regional reference values (WC) and values published by WHO (BMI percentile/SDS).g. Plasma samples for insulin, total IGF1 and IGF2, IGFBP1, -2, -3, -4, -6, and -7 analysis were stored at −80 °C until the time of the assay.Blood was taken morning in the fasting state. Blood concentrations of glucose, insulin, adiponectin, apelin, cholecystokinin, fibroblast growth factor 21, glucagon-like peptide-1, leptin, leptin receptor, resistin, and visfatin were measured at fasting as well as at 60 and 120 min of the standard oral glucose tolerance test (OGTT) performed using 1.75 g of anhydrous glucose per kilogram of body weight (maximum of 75 g). Collection was performed once after selection to the study. The samples were collected in tubes containing aprotinin. The material was immediately delivered to the laboratory at +4 °C and centrifuged for 15 min at a relative centrifugal force of 1590× IGF1—Labor Diagnostika Nord GmbH & Co.KG, Germany,IGF2—Mediagnost, Reultingen, Germany,IGFBP1, IGFBP2, IGFBP3, IGFBP6—Mediagnost, Reultingen, Germany,IGFBP4, IGFBP7—FineTest, Wuhan, China.Fasting insulin concentrations were measured with immunoradiometric kits (BioSource Company Europe S.A). The concentrations of total IGF and BP were measured using ELISA kits as follows:We assessed the whole genome expression in peripheral blood leukocytes using GeneChip Human Gene 1.0 ST Array . Total RNA extraction was performed using RiboPure Blood Kit . The whole transcript microarray experiment was performed according to the manufacturer’s protocol .t-test  or Mann–Whitney test  were used. Two-sided p-values < 0.05 were considered statistically significant. To assess the correlations between 2 continuous variables, Spearman’s rank correlation coefficient was calculated. Two-sided p values < 0.05 were considered statistically significant. Gene expression data were robust multi-array average (RMA)-normalized and presented as mean and standard deviation. ANOVA was used to examine the differences in gene expression between 2 independent groups. Benjamini–Hochberg (B–H)-corrected p values < 0.05 were considered statistically significant. Statistical analysis was performed using Statistica 13.3 .Continuous clinical and biochemical variables were presented as mean or median as appropriate. The Shapiro–Wilk test was used to assess the normality of continuous variables. To examine the differences between two independent groups, the Student’s The Permanent Ethical Committee for Clinical Studies of the Medical College of the Jagiellonian University approved the study protocol (consent number KBET/249/B/2013 26 October 2013). All parents, adolescent patients, and adult patients signed a written informed consent before blood sample collection. The study conforms with The Code of Ethics of the World Medical Association (Declaration of Helsinki), printed in the British Medical Journal (18 July 1964).The characteristics of study group was presented in p < 0.05) for IGFBP4; there was a trend for IGFBP3. However, the median value of IGF2, IGFBP1, IGFBP2, IGFBP6, and IGFBP7 were lower in the OB group than in the HC group, and the differences were significant for all the parameters (p < 0.001) except for IGF2 and IGFBP7, where the trend was observed.The differences in the mean concentrations of the IGF-axis proteins were presented in The correlation results of IGF1 and IGF2 proteins’ concentrations with selected metabolic parameters are presented in Fibroblast growth factor 21 was positively correlated with IGFBP6. Ghrelin was positively correlated with IGF2, IGFBP1, and IGFBP2 and negatively with IGFBP4. Leptin was negatively correlated with IGF2, IGFBP1, and IGFBP2. The leptin receptor was positively correlated with IGFBP1, IGFBP2, and IGFBP6 and negatively with IGFBP3. For IGF2, the correlation was positive but not significant. The level of resistin in the 60th and 120th minute correlated positively with IGFBP2 and negatively with IGFBP3. Visfatin revealed a positive correlation with IGFBP4, IGFBP6, and IGFBP7.p = 0.023), and the expression of IGF2 (p = 0.037) and IGFBP1 (p = 0.046) genes were lower in the OB group. IGFBP6 gene expression tended to be lower (p = 0.059) in the OB group.The expressions of IGF proteins’ genes are presented in In our study, we showed that the concentration of IGF-axis proteins differs in the healthy and obese pediatric population, as well as numerous, statistically significant correlations between the concentration of the studied proteins and the concentration of adipokines, gastrointestinal tract hormones, insulin or blood pressure. Additionally, we showed statistically significant differences in gene expression of IGF proteins’ family between obese and healthy children.In the study, we noted the significant differences in IGF-axis proteins’ concentration between the group of obese and healthy children, which may play a role in the pathogenesis of obesity. IGFBP4 showed significantly higher values, and there was a trend toward a higher concentration of IGFBP3 in the OB compared to the HC, while IGFBP1, IGFBP2, and IGFBP6 achieved significantly lower values, and IGFBP7 tended to be lower in OB. No significant difference in the median concentration of IGF1 protein in the serum of obese children and in healthy children was found. The currently published results are inconsistent showing higher , lower ,17,18,19p = 0.06) than in the control group. This protein plays an important role in the fetus growth regulation. Increased secretion is believed to be responsible for overgrowth of the fetus and increased amount of adipose tissue [In the group of obese children, the concentration of IGF2 was lower almost significantly (e tissue . Alfarese tissue . HoweverIGF binding proteins (IGFBPs) are a family of structurally similar proteins that are responsible for transport, extension of half-life, regulation of clearance, and direct modulation of IGF activity . IGFBP1 IGFBP3 is the most abundant protein in the serum and is responsible for the transport of 90–95% of IGF1 and IGF2 . This prIn our study, IGFBP4 had a significantly higher concentration in obese patients than in non-obese children. The liver is the main place of production of this protein, but its presence has also been demonstrated in other tissues ,40. IGFBIGFBP6 has a much greater affinity for IGF2 than for IGF1 . TherefoAdditionally, the IGFBPs undergo numerous post-translational modifications that may change their properties, and they are sensitive to the action of various types of proteases . It has In our study, the correlations of selected parameters related to obesity and other IGF-axis proteins were also assessed. All the correlations we describe is statistically significant. However, they differ in terms of strength  in the OB group. On the other hand, higher expression was noted in the case of IGFBP7 protein. At present, in the literature, there are no comprehensive data on the expression of the IGF-axis genes in humans. The standard source of diagnostic material is peripheral blood, which is readily available. Mononuclears show high expression of genes involved in lipid homeostasis and rapidly detect signals of its disturbance [Comparing the mRNA expression of the IGF-axis proteins, we obtained statistically significantly lower values among obese children of IGF2 and IGFBP1; IGFBP6 tended to be lower (turbance . These gThe study carried out in rats regarding IGF1 expression showed different results of the intensity of expression in obese individuals depending on the type of tissue . AnotherA study in the adult population showed higher levels of IGFBP2 and IGFBP7 mRNA in obese subjects, and lower IGFBP4 mRNA expression. In our study, despite lower IGF-1 mRNA expression in obese children, we showed no significant difference in IGF-1 protein concentration. The decreased expression of IGF2, IGFBP1, and IGFBP6 corresponds to the decreased amount of these proteins in the group of obese children. In the case of decreased IGFBP7 concentration, we showed higher IGFBP7 mRNA expression. The above results indicate that the regulation of the transcription may be of greater importance for the level of IGF2, IGFBP1, and IGFBP6 than for other components of IGF axis. However, the mechanism of regulation of the concentration of proteins of the IGF family seems to be complex and, apart from epigenetic factors, also influenced by many additional factors.There are few data in the literature that attempt to describe the relationship between obesity and IGF-axis proteins, e.g., in the papers by Saitoh et al. or Ballerini et al. ,72. HoweThe main limitation of the study was the small sample size, which could affect the validity of the results. In the future, it would be advisable to carry out similar experiments on larger cohorts to further confirm the obtained results. Certain outcomes, which did not reach the statistical significance cutoff value, are likely to be found significant when investigated with a population of greater size.The study revealed relationship between IGF–axis proteins and gene expression and childhood obesity with its metabolic parameters. We suggest that the IGF axis may be involved in obesity development, but the exact mechanism cannot be distinctly defined based on the study. The relationship between GH, IGF, and IGFBPs, as well as their interactions with the body’s cells, is complex and remains not fully understood. We found numerous correlations between IGFBP6 concentration and obesity metabolic parameters. As the available data for the expression and concentration of IGF-family proteins are inconsistent, further research, concerning pediatric obesity, including larger populations, is necessary.


Sanvitalia procumbens (creeping zinnia) in which the phytochemicals present in plant extract act as a stabilizing and reducing agent. For the stability of the synthesized AgNPs, different parameters like AgNO3 concentration, volume ratios of AgNO3, temperature, pH, and contact time were studied. Further, AgNPs were characterized by UV–visible spectroscopy, FT-IR (Fourier Transform Infrared Spectroscopy), XRD (X-ray Diffraction), SEM (Scanning Electron Microscopy), and EDX (Energy Dispersive X-ray Spectrometer) analysis. FT-IR analysis showed that the plant extract contained essential functional groups like O–H stretching of carboxylic acid, N–H stretching of secondary amides, and C–N stretching of aromatic amines, and C–O indicates the vibration of alcohol, ester, and carboxylic acid that facilitated in the green synthesis of AgNPs. The crystalline nature of synthesized AgNPs was confirmed by XRD, while the elemental composition of AgNPs was detected by energy dispersive X-ray analysis (EDX). SEM studies showed the mean particle diameter of silver nanoparticles. The synthesized AgNPs were used for photocatalytic degradation of Orange G and Direct blue-15 (OG and DB-15), which were analyzed by UV-visible spectroscopy. Maximum degradation percentage of OG and DB-15 azo dyes was observed, without any significant silver leaching, thereby signifying notable photocatalytic properties of AgNPs.Green synthesis of silver nanoparticles (AgNPs) employing an aqueous plant extract has emerged as a viable eco-friendly method. The aim of the study was to synthesize AgNPs by using plant extract of  Andrographis paniculata [Cathranthus roseus [Rhynchosia suaveolens [Metal nanoparticles have a variety of features in various fields. Since the sizes, dimensions, and compositions of metallic nanomaterials are all tied to their optical, physical and chemical properties, nanoscale materials have already been used in a wide range of applications . Nanostrniculata , Cathrans roseus  and Rhynaveolens , which aPhaseolus vulgaris have recently been shown to degrade 4-nitrophenol, reactive red 141 dyes [Solanum surattense, which are utilized as an efficient catalyst towards Rhodamine B (RhB) [Recently, silver nanoparticles (AgNPs) have risen in prominence due to their wide range of applications in biomedicine such as anticancer , antibac141 dyes , and oth B (RhB) . Green sSanvitalia procumbens (S. procumbens) aqueous plant extract, and then to explore the photocatalytic applications of the synthesized AgNPs. The synthesis of AgNPs using aqueous extract of this plant and its photocatalytic degradation of azo dyes (Orange G and Direct blue-15) has not been reported in literature, and in this article we are reporting it for the first time. According to an ethnobotanical survey of weeds in the state of Mexico, S. procumbens  is one of the major plant families of commercial importance for medicinal, herbal, culinary, and ornamental uses [S. procumbens aqueous plant extract has since been used as a possible bioreducing agent for the green synthesis of AgNPs in such a cost-effective and environmentally sustainable manner. In light of the foregoing, the current study was conducted to produce low-cost, clear, and easily reusable AgNPs from S. procumbens that contains bioreductants and stabilizers such as alkamides, phenolics, proteins, and terpenoids, which are responsible for the reduction of Ag+ to Ag0 [Sanvitalia procumbens-based silver nanoparticles can be highly reproducible. Large-scale utilization assessing the reproducible process can show the natural variability that occurs in the chemical composition of plant species that grow at the month of December, as well as assess how a plant can be used for this purpose so that the entire plant extract can be gainfully utilized in the process. Furthermore, the elemental composition, geometry, morphological, and optical study of synthesized AgNPs was carried out by using various techniques such as Energy Dispersive X-ray analysis (EDX), X-ray Diffractometer, Scanning Electron Microscope (SEM), Fourier Transform Infrared Spectroscopy (FT-IR), and UV–visible spectrophotometry. Moreover, the current study also assessed potential applications in the environment via green-synthesized AgNPs for the photocatalytic degradation of hazardous Orange G and Direct Blue-15 azo dyes by considering these dyes to be biodegradable.In the light of other discrepancies, different biological methods have reported on the synthesis of AgNPs and on the photocatalytic activities of other dyes. So, this work was carried out to synthesize AgNPs using tal uses . Thus, t+ to Ag0 ,26,27,28S. procumbens with the AgNO3 solution at a ratio of 1:9 (0.01 mol/L solution of AgNO3), the color change indicated that the synthesis of AgNPs occurred, where the plant extract was used as a reducing and stabilizing agent in this analysis. According to the literature, the initial indication of AgNPs was observed as a color transition from pale yellow to dark yellow and eventually colloidal brown, within a few minutes. The observed results were consistent with previous reports in which AgNPs were synthesized using various plant extracts, and aqueous silver nitrate solution (10−3 mol/L) altered the color from transparent to brown after extract addition [Several studies have stressed and illustrated the possible use of shade-dried plants as a reducing and stabilizing source for nanoparticle synthesis. After integrating an aqueous plant extract of addition ,30.3 solution was observed in a UV-visible spectrophotometer. However, the synthesis of AgNPs was confirmed by a mixture of AgNO3 and plant extract. The AgNPs formation in aqueous colloidal solution showed the sharp SPR (surface plasmon resonance) peak recorded at 438 nm as shown in Datura stramonium plant extract was used to reduce silver nanoparticles, and UV-visible absorbance spectra revealed that the SPR (surface plasmon resonance) band for silver occurred in the 400–450 nm range. The reproducibility of AgNPs synthesis was confirmed using the aqueous extract of a previously collected batch of plant, which provided silver nanoparticles when the optimized ratio of plant extract and AgNO3 (1:9 mL) was used [On the nanoscale level, one of the most intriguing features of metal nanoparticles is their optical properties, which vary proportionally with their size and shape. The large-size AgNPs are obtained at the weak absorbance peak and shift towards higher wavelength, while maximum absorbance peak shifts towards a lower wavelength when the particle size becomes smaller. In the current study, the absence of an absorbance peak in the visible region was examined when the plant extract and AgNOwas used ,34.3 concentrations under ideal circumstances. The size and forms of AgNPs were affected by varying the concentration of AgNO3 solution. In this analysis, SPR (surface plasmon resonance) peak showed that at lower AgNO3 concentrations, AgNPs with larger size distributions were present, implying the presence of AgNPs with broader size distributions. The SPR peak became sharper and narrower as the concentration of AgNO3 rose, along with an increase in its intensity [3 solution was increased. However, if the concentration of AgNO3 further increases to 0.02 mol/L, the intensity of such SPR peak weakens and broadens due to agglomerations induced by Van der Waals interactions between adjacent nanoparticles. It was concluded that with 0.01 mol/L AgNO3 solution, a distinct peak was found at 438 nm, but as stated in the literature, the concentration rises from 0.01 mol/L, and the intensity peak of synthesized nanoparticles decrease, which was also mentioned in This parameter was measured by varying AgNOntensity . This su3v/v ratios were used. The transition in spectrum was observed at each ratio, as seen in 3 solution, as mentioned in 3 solution, the peak intensity also increased, indicating that small-size AgNPs were obtained. While at lower volume of AgNO3 solution, non-uniform large-sized AgNPs were synthesized [v/v ratio, beyond this the peak becomes broader. The absorbance showed the quantity of AgNPs formed due to the reduction of silver ions.To decide the impact of volume ratios on AgNPs, different extracts and AgNOthesized . As a reThe production of AgNPs is heavily influenced by temperature. The reaction mixture of synthesized AgNPs was placed at various temperatures from 20–70 °C for 30 min. The color and intensity shift of the reaction mixture is illustrated in 3 solution at different time intervals. The AgNO3 solution reacts with the plant extract, resulting in a color shift within 1 h of the reaction. With the aging process, the color became more intense [+ to Ag0). Since a significant amount of Ag+ was transformed to Ag0, an outstanding SPR (surface plasmon resonance) spectrum was found as the reaction time increased. The absorption peak of synthesized AgNPs was observed in the UV–vis spectra with strong SPR in the range of 438 nm at 8 h, indicating fast synthesis  spectroscopy. The data suggests that during AgNPs synthesis, the phytochemicals present in a plant extract play an important role in the production of metal nanoparticles, which reduce the metal salt into its nanoparticles. It is tough to select an individual phytochemical entity, but in general, flavonoids, phenolic groups, organic acids, and proteins all are thought to be essential for the synthesis of nanoparticles [S. procumbens extract showed prominent bands of absorbance at around 3277, 2923, 2364, 1590, 1405, and 1033 cm−1 , (200), and (220) Bragg reflections, respectively, which were the exact peak positions as given for the face center cubic (FCC) lattice structure of AgNPs. This diffraction pattern was very similar to JCPDS file no. 04–0783 standard diffraction pattern [D is crystallite size, λ corresponds to the X-rays wavelength that used 1.5406 °A, β is FWHM , and θ corresponds to Bragg’s angle.To investigate the crystalline nature and surface morphology of the synthesized AgNPs, the X-ray diffraction (XRD) analysis was performed in the range of 20–70 degrees at 2θ. The XRD pattern of the AgNPs synthesized by the reaction of silver salt solution with an aqueous plant extract of  pattern . The 38° pattern . The parThe elemental composition was determined by using energy dispersive X-ray (EDX) analysis. The EDX result of synthesized AgNPs revealed the existence of silver, as well as the residual carbon, oxygen, and chlorine peaks that were unwittingly inserted from the surface molecule of the plant extract. Green-synthesized AgNPs were responsible for the observed C, O, Cl, and Ag peaks. The peak of Ag at 3 eV confirms the formation of silver nanoparticles, where standard weight percent of silver was 90.88%. In this analysis, the green-synthesized AgNPs showed aggregated morphologies and a nonuniform size distribution . The parResearchers used scanning electron microscopy (SEM) to design the images of the sample and consider the morphology of the nanoparticles. The shape and size of synthesized AgNPs by using green synthesis were shown in a micrograph of a scanning electron microscope (SEM). According to the given micrograph, the scale of the synthesized AgNPs was up to 250 nm. The micrograph revealed that the particle sizes range from 10–80 nm and are spherical in shape. Most of them are synchronized in the form of large clusters, but few of them are dispersed as seen in S. procumbens-AgNPs [To investigate the photocatalytic capability of stable AgNPs towards the degradation of various organic dye molecules, the commonly used Orange G dye in many dyeing industries was selected. Orange G is a well-known water-soluble dye that has always had widespread usage in industries such as leather, textiles, pharmaceuticals, paper, and printing . To evalns-AgNPs .Dye pollution has grown in tandem with the development of industries that use colorants, such as in clothing, leather, food, and agrochemicals. Since the majority of commonly used azo dyes are made from benzidine, these dyes contain a carcinogenic substance that is hazardous to marine biota. The azo dye DB-15 used in the textile had a variety of toxic effects on microalgae, Cladocera, and zebrafish embryos. So, the dumping of this colorant waterbody should be controlled to prevent environmental impacts . As a reS. procumbens plant was harvested from the area of Kohat, Khyber Pakhtunkhwa, Pakistan and was identified by Dr. Nisar Ahmed, Department of Plants and Environmental Sciences, Kohat University of Science & Technology.The S. procumbens was washed thoroughly with tap water to remove debris and other contamination, followed by washing with distilled water. The plant was shade-dried at room temperature. Twenty grams of finely cut plant was heated at 70 °C for 30 min in 200 mL of distilled water. The extract was cooled down and then filtered via Whatman No. 1 filter paper, and the aqueous plant extract was stored in the refrigerator for further analysis.The plant of 3 solution in a dropwise manner. After mixing the aqueous plant extract into silver ion solution, the reaction mixture was placed onto a rotatory orbital shaker operating at 200 rpm, then it was subjected to a water bath for 70 °C. Complete reduction of Ag+ to Ag0 ions in the reaction mixture was confirmed by the color change of AgNO3; plant extract solution from pale yellow to dark brown implied the formation of AgNPs (further scanned by UV-visible spectrophotometry). Finally, the samples were centrifuged for 15 min at 6000 rpm. The organic moieties were removed after centrifugation, and the pellet was washed with distilled water and methanol. The powder form of nanoparticles was obtained, which were utilized for further characterization and photocatalytic studies.At room temperature, 1 mL volume of aqueous plant extract was added to 9 mL of 0.01 mol/L AgNOS. procumbens aqueous plant extract and different parameters including AgNO3 concentration, volume, pH, time, and temperature were studied to check the stability of AgNPs. Spectrophotometric study of AgNPs after 6–8 h of incubation in the dark at room temperature revealed the optimal conditions for efficient reduction of silver ions and eventual nucleation into silver nanoparticles. The impact of plant extract volume on green synthesis of AgNPs was explored through executing the reaction with various volumes of AgNO3 solution , whilst maintaining the volume of plant extract remains as constant. The reaction mixtures were deposited with various AgNO3 concentrations, which were used to investigate the impact of AgNO3 concentration on AgNPs synthesis . So, the optimal AgNO3 concentration measured the absorbance of nanoparticles by UV-visible spectrophotometer. The aforementioned reaction mixture was subjected at different temperature conditions, namely 20 °C, 30 °C, 40 °C, 50 °C, 60 °C, and 70 °C, for synthesizing AgNPs. Following the synthesis, every single reaction tube was held at the specified temperature for 15 min. The pH effect was observed by exposing a reaction mixture of AgNPs at various pH level, and 0.1 mol/L NaOH solution was used to change the pH of the reaction solution to the optimal level.Green synthesized AgNPs were produced by using −1 resolution, within the range of 4000 to 500 cm−1 [To calculate the reduction of silver nitrate into silver nanoparticles, UV-visible spectroscopy was used on a regular basis (Shimadzu-UV-1800). The AgNPs sample was diluted with distilled water and then examined under UV-visible spectrophotometer. The quartz cuvette cell filled with distilled water was used as a reference for recording the spectrograph at scanning speed of 300–700 nm . The mor500 cm−1 ,59.2 = 0.9935) and y = x0.034 − 0.0428 (R2 = 9.997) for OG and DB dyes, respectively.0’ is the initial absorbance of Orange G and DB-15 azo dyes, and ‘A’ shows the absorbance of dyes solution obtained at each time interval [The photocatalytic degradation studies were performed by mixing 100 mL of the reaction mixture into a 250 mL glass beaker, distinctly. Prior to being irradiated, the reaction sample (0.05 g/100 mL) was mixed with (15 ppm) OG and (10 ppm) DB-15 azo dye solution, respectively. Both solutions were constantly stirred for 30 min in the dark to ensure that equilibrium was reached. An absorption capacity of pure AgNPs for OG was evaluated at room temperature in the dark condition. Then the solution was irradiated by a Hg lamp with a high-pressure  sealed in to a rectangular steel box. The reactors were put on a magnetic stirrer to ensure that the solvent was evenly dispersed. After the irradiation times, aliquots of a previously utilized sample were removed at specific time intervals. Finally, the solution was centrifuged for 10 min at 4000 rpm to extract the suspended particles and then examined with a UV–visible spectrophotometer. The experiment was repeated twice, and the standard deviation did not reach 5%, indicating that the findings were relevant. The percentage degradation was calculated by using the following equation on the base of standard curve y = x0.0386 − 0.035 (RS. procumbens. The synthesized AgNPs were successfully characterized by using different techniques like UV-Visible, FT-IR, SEM, XRD, and EDX. Furthermore, AgNPs exhibited potential photocatalytic degradation activity of Orange G and Direct blue-15; the synthesized catalyst showed more degradation efficiency against OG dye than DB-15. Overall, all these results suggest that AgNPs, as synthesized in this study, can be used as a potent photodegradation analyte.In the current study, the synthesis and optimization of AgNPs was demonstrated by using plant aqueous extract of