The aim of this study was to better understand the characteristics and etiology of acute nonvariceal upper gastrointestinal bleeding (ANVUGIB) in recent years in this region and to provide evidence-based medical evidence.  100 patients with acute nonvariceal upper gastrointestinal bleeding (ANVUGIB) who met the clinical diagnostic criteria of ANVUGIB admitted to Suzhou First People's Hospital from January 2017 to December 2021 were analyzed, as well as the age difference and change rule. According to age, 100 patients were divided into young (18-39 years), middle-aged (40-59 years), and elderly (60 years and above), and the differences in the three groups were compared. The etiology was confirmed by endoscopic examination and was recorded one by one in a well-designed ANVUGIB case data registration form. Statistical software SPSS 23.0 was used for analysis. P < 0.01). Drinking, fatigue, and emotional excitement led to a higher proportion in the middle-aged group and the young group, in comparison to the elderly group (P < 0.01).  Gastric ulcer was the main cause in the elderly group (50.0%), duodenal ulcer was the main cause in the middle and young groups, and gastrointestinal cancer (7.1%) and marginal ulcer (2.3%) in the elderly group were higher than those in the young group. Nonsteroidal anti-inflammatory drugs (52.3%) were the main inducement in the elderly group, which was significantly higher than in the middle-aged group (13.1%) and the young group (5%) ( Peptic ulcer is the most common cause of acute nonvariceal upper gastrointestinal bleeding, followed by acute gastric mucosal lesions and upper digestive system tumors, compared with nonulcer. Acute nonvariceal upper gastrointestinal bleeding (ANVUGIB) refers to bleeding caused by nonvariceal diseases of the digestive tract above the flexion ligament, including bleeding from the pancreas and biliary tract and bleeding caused by diseases near the anastomosis after gastrojejunostomy, with an annual incidence of 50~150/100,000, and a case fatality rate of 6%~10% . TherefoTherefore, the clinical characteristics of ANVUGIB, such as inducing factors, etiology composition, and onset age, have undergone corresponding changes . It is oWith the continuous development and improvement of endoscopic treatment technology, the advantages of endoscopic treatment for acute nonvariceal upper gastrointestinal bleeding are becoming more and more obvious, which greatly reduces the mortality, rebleeding rate, and surgical treatment rate of bleeding patients, saves resources and reduces costs, and opens a new chapter for the treatment of ANVUGIB –19. TherThis study will carry out a large-sample clinical epidemiological study to clarify the etiological structure, variation, and influencing factors of acute nonvariceal upper gastrointestinal bleeding, in order to clarify the etiology for improving prognosis and guiding treatment.Medical records of a total of 100 inpatients who met the clinical diagnostic criteria of ANVUGIB and had the etiology confirmed by endoscopic examination from January 2017 to December 2021 were included. According to age, the 100 patients were divided into young (18-39 years), middle-aged (40-59 years), and elderly (60 years and above), and the differences in the three groups were compared. Age ranged from 18 to 96 years, with an average of 54.6 ± 16.3 years. There were 80 males and 20 females, with a male to female ratio of about 4:1.(1) The patient presents symptoms of hematemesis and/or melena, which may be accompanied by dizziness, pallor, increased heart rate, decreased blood pressure, and other signs of peripheral circulatory failure, and the diagnosis of acute upper gastrointestinal bleeding is established. (2) Endoscopic examination found nonvariceal bleeding lesions in the upper digestive tract, and ANVUGIB diagnosis could be established.(1) We exclude patients with bleeding from lesions of the mouth, nose, pharynx, or respiratory tract. (2) We exclude patients who are taking some drugs (such as iron and bismuth) and food  or have black stool. For suspicious patients, gastric juice, vomit, or fecal occult blood test can be performed. (3) Lower gastrointestinal bleeding and bleeding caused by esophagogastric varices were excluded. (4) Patients younger than 18 years old and without endoscopic examination to determine the cause of bleeding were excluded.Calibration criteria for successful hemostasis: after treatment, the bleeding was stopped under gastroscopy, the patient's condition was stable, vital signs were stable, there were no signs of active bleeding and no hematemesis and large amounts of melena, the stool gradually turned yellow, and stool specimens was negative. [Criteria for rebleeding: one of the following conditions occurs after the first successful hemostasis treatment: hematemesis, melena/hematochezia, the stomach tube sucks out fresh blood, hemodynamic instability , and rebleeding was confirmed under endoscopy. [Criteria for the effectiveness of endoscopic treatment: the first endoscopic treatment was successful and there was no further bleeding within 3 days ((1)egative.  Criteriadoscopy.  Criterian 3 days .The design scheme is as follows: well-designed ANVUGIB case data registration form was signed, recording The general data of 100 ANVUGIB patients  and date of visit were recorded by details; while also contained inducement before onset and Rockall score; and treatment method, endoscopic time, endoscopic diagnosis, Forrest grade, and disease outcome of patients .P values < 0.05 were considered significant.Continuous data are expressed as the means ± SDs. All statistical calculations were carried out using SPSS 23.0 software. A chi-square test was used to compare count data and groups. For multiple comparisons, data were analyzed via analysis of variance (ANOVA) with the Tukey-Kramer Multiple Comparison Test. There were 100 patients, 80 men and 20 women, with a male to female ratio of approximately 4 : 1. Age 18~96 years, with a mean of 54.6 ± 16.3 years, included 42 elderly patients aged 60 years, 38 middle-aged patients aged 40~59 years, and 20 young patients aged 18-39 years.After admission, all patients were given symptomatic treatment such as fasting, fluid rehydration, and acid preparation. For patients with serious conditions and hemodynamic changes, supportive treatments such as dilatation, blood transfusion, and correction of shock should be performed to maintain blood pressure stability and correct hemoglobin to more than 70 g/L. An endoscopic examination was performed as soon as possible, and an electronic gastroscope was used for the operation. ECG monitoring and high flow oxygen inhalation (oxygen flow 4-5 L/min) were performed during the examination. The etiology was confirmed by endoscopic diagnosis, and endoscopic treatment was performed according to the needs of the disease. Among the 100 patients, 2 cases had rebleeding after admission. None of the patients died in the hospital because seriously ill patients were discharged automatically,. And 2 gastric cancer bleeding cases were transferred to surgical treatment. The 100 patients were followed up by telephone after discharge, and 2 patients had died, both of them died from non bleeding-related diseases, including 1 patient with advanced tumor consumption, and 1 patient with cardiovascular and cerebrovascular diseases.The results showed that peptic ulcer was mainly used in the three groups, with ulcer disease in the old group reaching 76.1% and 88.8% in the young group. Ulcer disease accounted for 86.8% of the middle-aged group .P < 0.01, and for the middle and others, the main causes were drinking and fatigue, which was significantly higher than for the elderly group (P < 0.01). Overall, there were no significant differences between NSAIDs, drinking, and exertion among the three major triggers (P > 0.05). ALL patients were common in acute nonvariceal upper gastrointestinal bleeding predisposition , which was significantly higher than in the middle-aged group (13.1%) and young group (5%), position .P < 0.05, statistically significant difference). In the combined treatment group, 91.3% of grade IA and grade IIA lesions were treated by titanium clip combined with submucosal epinephrine injection. Therefore, mechanical clamping of the titanium clip is the main treatment method for grade IA lesions and grade IIA lesions, and the immunohistochemistry of gastrointestinal bleeding tissue is shown in Analyzing the treatment efficiency of different treatment groups with different Forrest grades, we show that, overall, grade Forrest II lesions are more efficient than each treatment method of grade Forrest lesions. For grade Forrest Ia and A lesions, titanium clamp and combination treatment were highly efficient, significantly higher than in the injection and APC electrocoagulation groups (ANVUGIB is mainly caused by diseases of the upper digestive tract and biliary and pancreatic diseases. Peptic ulcers, upper gastrointestinal tumor, and acute gastric mucosal lesions were the most common . This stIn addition to ulcer disease, the upper gastrointestinal tumor was the second cause of acute nonvariceal upper gastrointestinal bleeding . Due to The advantage of this study was to demonstrate the importance of ulcer in the progress of nonvariceal upper gastrointestinal bleeding, which was brought to light for the future prevention and treatment of this disease. However, there are also limits to this study. First, the number of patients is not so large, which needs further validation in future. Second, the mechanism was not clarified. Further studies are needed to carry more experiments to explain it.Peptic ulcer is the most common cause of acute nonvariceal upper gastrointestinal bleeding, followed by acute gastric mucosal lesions and upper digestive system tumors, compared with nonulcers. Among the causes of acute nonvariceal upper gastrointestinal bleeding, gastric ulcers were mainly found in the elderly group, while duodenal ulcers in the middle and young groups, and the proportion of tumors in the elderly group was higher than that in the younger group.


Bronchogenic cysts are congenital malformations caused by aberrant foregut budding. They major occur in the thorax, with subdiaphragmatic cases being uncommon. Here, we present a series of 19 patients diagnosed with subdiaphragmatic bronchogenic cysts histopathologically at a single institution in China from 2012 to 2021. A literature review was also conducted by searching the PubMed database using keywords related to “bronchogenic cysts” and “subdiaphragmatic,” yielding 107 cases. Taken together, the 126 cases had a median age of 41.0 years  and 62 of them were male (49.2%). The cysts were most commonly detected in the left adrenal region (36.2%), followed by the pancreatic region (11.5%) and gastric cardia/lesser curvature of the stomach (9.2%). All patients except two underwent surgery for a definite diagnosis, symptom alleviation, and (or) malignancy prevention. Most patients recovered fast and were discharged from the hospital within 1 week after surgery, and the surgical complications were infrequent. The prognosis was generally favorable, as no recurrence was reported during the follow-up as long as 77 months. Bronchogenic cysts are congenital cysts caused by aberrant budding of the primitive foregut or tracheobronchial tree. They usually locate in the thorax, especially in the mediastinum –3. HowevDespite the fact that cases of sBCs have been previously reported , 7, the All cases of sBCs from 2012 to 2021 at Peking Union Medical College Hospital in China were retrospectively identified from the hospital information system. Only patients with histopathological diagnoses of sBCs were included in the analysis. Parameters, such as demographic characteristics, clinical presentation, imaging findings, surgical information, and histopathological examination, were extracted from electronic medical records. All patients were contacted by phone in May 2022 for information on recurrence and surgical complications.t-test or Pearson’s chi-square test was employed when appropriate. All statistical analyses were performed using SPSS 22.0.For the literature review, PubMed was searched in May 2022 using the following keywords: “bronchogenic cyst*”, “bronchial cyst*,” “subdiaphragm*,” “retroperitoneal,” and “abdom*.” The detailed search strategy can be found in n = 12) or a radiographic examination done for unrelated indications (n = 2). Among the symptomatic patients (n = 5), two complained of flank pain, one of abdominal pain, one of coughing, and the other of acid reflux and abdominal distention. In all patients, the cyst was solitary with a median diameter of 5.1 cm (interquartile range [IQR]: 3.6-6.6 cm). The cysts were inclined to locate in the left abdomen  rather than the midline or right abdomen, the upper abdomen  instead of the lower abdomen, and the retroperitoneal space  compared with intra-abdomen. The left adrenal region  was the most common location, followed by the pancreatic region  and gastric cardia/lesser curvature of the stomach . On computed tomography (CT), the density of cysts ranged from low to slightly high. The number of cysts with low to slightly low, soft-tissue, and slightly high density on CT is eight, nine, and two, respectively. Only three patients underwent magnetic resonance imaging (MRI), and all of their cysts were hypointense on T1 weighted image. As for T2 weighted image, two of them were hyperintense while the other was hypointense. Twelve of 19 cysts (63.2%) showed no enhancement on CT or MRI. Ten patients received the B ultrasound examination, displaying an anechoic (n = 3), hypoechoic (n = 3), or mix-echoic (anechoic to hypoechoic) cyst (n = 4). Three of the cysts were multilocular with septa present. The representative appearance of the cysts on CT, MRI, and ultrasound is demonstrated in Nineteen patients  with histopathological diagnoses of subdiaphragmatic bronchogenic cysts from 2012 until 2021 were identified. Patient demographics are depicted in Laparoscopic surgery was conducted in most of the patients , while two underwent laparotomy and one underwent video-assisted thoracoscopic surgery VATS; . The conWe attempted to contact all patients for information on recurrence and long-term complications in May, 2022, with 12 responses. Unfortunately, not every patient had regular imaging examinations after surgery, but they all claimed no signs of recurrence until their most recent imaging assessment, and the longest one was 77 months after surgery. As for long-term complications, two patients (cases 8 and 12) had incision hernias and both received surgery for repairment. Additionally, left renal artery stenosis and left renal atrophy were discovered 8 months after surgery in case 5 whose cyst was located in the left adrenal region. Later, this patient received a left nephrectomy.194 records were retrieved initially after searching the PubMed using the strategy displayed in The 95 citations reported 107 cases  in total . The medsBC was solitary in 96.3% (103/107) cases. One case had two cysts in the left adrenal region , and oneGenerally speaking, patients with sBCs usually showed normal blood tests, though abnormalities could be seen seldomly. For instance, several patients had significantly elevated serum levels of carbohydrate antigen 19-9 (CA19-9), even reaching 4330.7 U/mL. In these cases, bronchial epithelial cells were positive for CA19-9 in the immunohistochemical study on surgical specimens and the serum levels returned to normal after surgery , 81, 98.All patients except two underwent surgery in the literature. Laparoscopy was the most common surgical procedure for patients with sBCs , followed by laparotomy (23/72 31.9%). One of the two patients that did not receive surgery was diagnosed with sBC based on the pathohistological examination of the specimen gained from CT-guided biopsy . The patBronchogenic cysts are primitive-foregut-derived congenital cystic abnormalities that usually occur in the thorax, particularly in the mediastinum –3. InfreAs displayed in three articles, sBCs can be detected even during pregnancy , 64, 87,Streptococcus anginosus, providing direct evidence of intra-cyst infection. Patients with sBCs lacked specific symptoms, which was the same for laboratory findings. The tumor markers test did not aid in the diagnosis of sBCs, as nearly all patients tested negative for them. Seldomly, the epithelial cells lining the cyst were able to produce tumor markers like CA19-9, leading to a significantly elevated serum level . Symptoms caused by sBCs were usually non-specific, mainly due to complications such as local compression and infection. For example, one patient with sBC in the left adrenal region had a 5-year history of hypertension . After tum level , 81, 98.Up to now, a histopathological examination is required for the diagnosis of sBCs. Biopsy guided by CT or EUS could provide the specimen before surgery. Several cases from the literature were diagnosed based on biopsy, and two of them did not receive surgery because the patients were asymptomatic , 82. In n = 12 in our series, and n = 33 in the literature) patients had no recurrence during the follow-up, and the longest one reached 77 months in our series. Particularly, one patient with malignant change reported in the literature was also free of recurrence for 14 months after surgery (After the complete cyst(s) resection, patients with sBCs generally had a good prognosis. Forty-five ( surgery .In conclusion, our series added another 19 cases to the list of rarely reported sBCs. Together with the 107 published cases, we demonstrated that the sBCs are predisposed to locate in the left adrenal region, pancreatic region, and gastric cardia/lesser curvature of the stomach. Patients with sBCs lack specific clinical symptoms, laboratory examinations, and imaging features. Surgery is still the best management strategy for patients for a definite diagnosis, symptom alleviation, and complication prevention. The prognosis is generally favorable after complete resection of the lesion.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. The ethics committee waived the requirement of written informed consent for participation.WL conceived the study. JX and XZ contributed to the case collection, discussion, literature review, and first manuscript draft. WL, HZ, TH, BL, and XH provided critical revisions. All authors contributed to the article and approved the submitted version.


Phytocannabinoids possess a wide range of immune regulatory properties, mediated by the endocannabinoid system. Monocyte/macrophage innate immune cells express endocannabinoid receptors. Dysregulation of macrophage function is involved in the pathogenesis of different inflammatory diseases, including inflammatory bowel disease. In our research, we aimed to evaluate the effects of the phytocannabinoids D9 tetrahydrocannabinol (THC) and cannabidiol (CBD) on macrophage activation. Macrophages from young and aged C57BL/6 mice were activated in vitro in the presence of pure cannabinoids or cannabis extracts. The phenotype of the cells, nitric oxide (NO•) secretion, and cytokine secretion were examined. In addition, these treatments were administered to murine colitis model. The clinical statuses of mice, levels of colon infiltrating macrophages, and inflammatory cytokines in the blood, were evaluated. We demonstrated inhibition of macrophage NO• and cytokine secretion and significant effects on expression of cell surface molecules. In the murine model, clinical scores were improved and macrophage colon infiltration reduced following treatment. We identified higher activity of cannabis extracts as compared with pure cannabinoids. Each treatment had a unique effect on cytokine composition. Overall, our results establish that the effects of cannabinoid treatments differ. A better understanding of the reciprocal relationship between cannabinoids and immunity is essential to design targeted treatment strategies. Macrophages are specialized innate immune cells that orchestrate homeostatic, inflammatory, and reparative activities. Murine macrophages are located in the brain, skin, liver, kidney, lungs, and heart and originate from the yolk sac or fetal liver; their maintenance in adulthood in the absence of stressors is independent of circulating monocytic precursors. In other tissues, such as the gastrointestinal tract, monocytic precursors contribute to tissue macrophages . In the Immunological dysregulation in IBD is characterized by epithelial damage, expansion of inflammation driven by intestinal flora, a large number of cells infiltrating into the lamina propria, and a failure of immune regulation to control the inflammatory response . In IBD The endocannabinoid system (ECS) regulates various aspects of physiological, behavioral, immunological, and metabolic functions. It is now clear that many of the components of the endocannabinoid system function as key regulators of the immune system and the immune response . EndocanPhytocannabinoids, the biologically active constituents of cannabis, possess a wide range of immune regulatory properties, mediated by the endocannabinoid system. Two cannabinoids have been the focus of most of the studies that have examined medical uses, i.e., D9 tetrahydrocannabinol (THC) and cannabidiol (CBD). THC and some of the other phytocannabinoids mediate their biological effects primarily through the classical cannabinoid receptors CB1 and CB2. In addition, THC can act as an agonist of the receptors/channels GPR55, GPR18, PPARγ, transient TRPA1, TRPV2, TRPV3, and TRPV4, and as an antagonist of the receptors/channels TRPM8 and 5-HT3A. Interestingly, although CBD affects the immune function, it has a very weak affinity to CB2 or CB1, where it can act as a negative allosteric modulator. Several reports have demonstrated that CBD acts as an agonist of other receptors/channels, such as TRPA1, TRPV1, TRPV2, TRPV3, PPARγ, and 5-HT1A, and as an antagonist of the receptors GPR55, GPR18, and 5-HT3A. CBD is also an inverse agonist of the receptors GPR3, GPR6, and GPR12 [Previously, we compared the influence of cannabinoid-based treatments on lymphocyte function . The aimWhile Cannabis is not yet registered as a drug, the potential of cannabinoid-based medicines for the treatment of various conditions has led many countries to authorize their clinical use. As a result, in recent years, there has been a rapid increase in the medical use of cannabis and a wide range of cannabinoid-based treatments are offered to patients. THC and CBD are considered to be the two essential elements in these treatments. Therefore, it is crucial to explore the various biological effects of these molecules. A better understanding of the effects of THC, CBD, and other active molecules on the immune response will assist physicians in providing the best possible individually targeted treatment for their patients and will allow the design of new treatments.This research was performed under the approval of The Medical Cannabis Unit in the Israeli Ministry of Health (REQ46). Pure THC was generously provided by the laboratory of Prof. Raphael Mechoulam. Synthetic CBD was purchased from STI Pharmaceuticals Ltd., Newtown, UK. Cannabis Sativa and Indica extract with high content in THC or CBD  were supplied by Cannabliss . Extraction was obtained using ethanol, and evaporated. Identification and quantification of phytocannabinoids in the cannabis extracts were done by ultrahigh performance liquid chromatography with an ultraviolet detector (UHPLC/UV) system . The terpenoid analysis was performed by static headspace gas chromatography tandem mass spectrometry (SHS-GC/MS/MS) using full evaporation technique with external calibrations, as previously described ,16. The Female and male 8- to 11-week-old and 8-month-old C57BL/6 female mice were purchased from Envigo, Jerusalem, Israel and were acclimated for at least 7 days before the experiment in the specific pathogen-free (SPF) facility of the Authority of Biological and Biomedical Models at the Hebrew University of Jerusalem. The aged mice were up to 18 months old in the SPF animal facility. The study was approved by the Institutional Animal Care and Use Committee of the Hebrew University of Jerusalem in accordance with national laws and regulations for the protection of animals . The mice were housed under specific SPF conditions in the animal facility under the AAALAC accreditation, throughout the experiments.Peritoneal exudate cells were induced in mice by an intraperitoneal injection of 0.5 mL of 3% thioglycollate . After 4 days, mice were anesthetized with ketamine and xylazine, and then killed by cervical dislocation. Peritoneal exudate cells were washed from the peritoneal cavity of mice by lavage with 5 mL of ice-cold, sterile phosphate buffered saline (PBS). Cells were washed with PBS and re-suspended in Dulbecco’s modified Eagle medium (DMEM)  supplemented with 10% fetal calf serum (FCS), 1% penicillin/streptomycin, and 1% L-glutamine . Cell viability was determined by MTT colorimetric assay in which a yellow tetrazole, is reduced to purple formazan in living cells . The resultant color was measured at 450 nm using a Biotek PowerWave XS Microplate Reader.5 cells/well in 96-well plates and incubated overnight at 37 °C and 5% CO2. On the following day, the medium was changed to fresh DMEM containing 5 µg/mL CBD, THC, or cannabis extracts. The cells were then stimulated for 24 h by the addition of lipopolysaccharide (LPS) to a concentration of 1 μg/mL. After 24 h, cell supernatants (SNs) were harvested for nitric oxide radical (NO•) assay by addition of 100 μL SN to an equal volume of Griess reagent . After 10 min of incubation, the resultant color was measured at 550 nm. The amount of NO• produced, and any inhibition by the tested materials, was calculated from a standard curve prepared with NaNO2. Controls: non-activated cells, activated cells + vehicle, activated cells + 1400W dihydrochloride .Peritoneal macrophages were seeded at a density of 2.5 × 105 cells/sample were washed once in ice-cold staining buffer . Then, cells were stained in the dark at 4 °C for 30 min with fluorochrome-labeled anti-mouse mAb , specific for cell surface antigens: F4/80, I-Ad (MHC class II), and CD16/32. Cells were subsequently washed, re-suspended in staining buffer, and analyzed by flow cytometry.5 × 10Total cellular RNA was extracted using RNeasy Mini Kit columns , according to the manufacturer’s protocol. One microgram of total RNA was used to synthesize cDNA using a high-capacity cDNA kit , following the supplier’s instructions. Detection of transcript levels of CB1 and CB2 was performed using a TaqMan Gene Expression Assay Kit , with HPRT-1 as a reference. All primers were purchased from Applied Biosystems . Real-Time PCR reactions were conducted using a QuantStudio 5 instrument . Data were analyzed using the QuantStudio design and analysis Software .Colitis was induced in C57BL/6 mice with 2% DSS dissolved in drinking water given ad libitum (Days 1–7), and then replaced with plain drinking water for 3 days. Then, 5 mg/kg cannabis/cannabinoids were prepared in 5% Cremophor EL , 5% ethanol  in PBS, and 0.1 mL were administered IP every other day, starting from Day 1. Body weight and stool were monitored once a day. Changes of body weight are indicated as loss of baseline body weight . Clinical score (0–9) included: stool score (0–3), rectal score (0–3), and general clinical parameters . On the tenth day of colitis induction, blood was collected from the mouse tails into ethylenediaminetetraacetic acid (EDTA)-coated capillary tubes, and then the mice were anesthetized using ketamine and xylazine, and then killed by cervical dislocation. The intestines were excised, measured, and carefully rinsed with saline. Blood tubes were centrifuged at 1500 rpm, room temperature, for 5 min; plasma was collected and kept at −80 °C for cytokine and chemokine analysis.Colon tissue was fixed in 4% buffered formaldehyde  and embedded in paraffin. For histology, the sections were stained with H&E according to standard protocols. Histological scoring (0–9) was based on 3 parameters: Crypt damage (0–3), percent involvement (0–3), and damage to bowel wall structure (0–3).2O. Sections were treated with 3% H2O2 and antigens retrieved by incubation with 1 mg/mL pronase. Then, the samples were washed in PBS and blocked in CAS blocking reagent . The slides were stained with anti-F480 . Anti-rat IgG universal immune peroxidase polymer  was used as secondary antibody. Sections were incubated with Stable Peroxidase Substrate Buffer , washed with H2O, and analyzed on a BX41 microscope .For immunostaining, paraffin embedded sections were heated to 60 °C, deparaffinized using xylene, dehydrated using ethanol, and washed with HPeritoneal macrophages were activated for 24 h with LPS, in the presence of 5 µg/mL CBD, THC, or cannabis extracts. The supernatant was collected and analyzed using a LEGENDplex™ MU Macrophage/Microglia Panel cytokine array assay , according to the manufacturer’s instructions.Plasma samples from DSS model mice were analyzed using a LEGENDplex™ Mouse Inflammation Panel cytokine array , according to the manufacturer’s instructions.p value < 0.05 were considered statistically significant.Data from in vitro studies are represented as mean ± SE. The mean was calculated from the indicated number of experiments. The mean of triplicates from each experiment was used for this calculation. For statistical analysis of the macrophage NO• secretion experiments in female and male mice, we used Friedman test. For statistical analysis of the macrophage NO• secretion and cannabinoid receptors expression, macrophage cytokine secretion, and flow cytometry experiments in young and aged mice, we used the Mann–Whitney test. For statistical analysis of colon length, colon histopathology, macrophage infiltration to the colon, and blood cytokines in the DSS model experiments, we used the Kruskal–Wallis test. For statistical analysis of weight loss and clinical score in the DSS model experiments, we used the Kruskal–Wallis test of area under the curve (AUC). In all experiments, p < 0.0001) than the pure cannabinoids in female and male mice. A treatment with the combination of THC and CBD (2.5 µg/mL of each) was less effective than the pure cannabinoids . To test the effect of cannabinoid treatments on macrophage activation, NO• secretion was determined. Macrophages from previously thioglycollate (tg)-injected C57BL/6 mice were collected by peritoneal lavage, and then activated for 24 h with lipopolysaccharide (LPS), in the presence of cannabinoid treatments; 1400W dihydrochloride, a specific iNOS inhibitor, served as control. All treatments show dose dependent effect on NO• secretion . For ourabinoids .Next, we examined the influence of aging on the responsiveness of macrophages to cannabinoid treatments. For this aim, we obtained peritoneal macrophages from aged (18 months old) mice and compared their NO• secretion with macrophages from young (2 months old) mice. To examine the effect of the treatments on inflammatory cytokine/chemokine secretion from peritoneal macrophages, we collected the supernatant of 24 h LPS-activated cells and analyzed the levels of different cytokines using a LEGENDplex™ MU Macrophage/Microglia Panel cytokine array assay. IL6, TNF-alpha, CXCL2, and G-CSF levels in the culture media increased upon activation a–d. Our To learn more about CBD and THC molecular effects in macrophages, we tested cell surface expression of several molecules in the activated macrophages. Class II molecules of the major histocompatibility complex (MHCII) is upregulated on some polarized macrophage populations upon activation. We found that both THC and CBD treatments induce small, but significant, further elevation in MHCII expression a. R. A. We chose the murine colitis dextran sodium sulfate (DSS) model to compare the efficacy of the different cannabinoid treatments in macrophage-related inflammation in vivo. DSS is a chemical colitogen with anticoagulant properties. In the acute intestinal inflammation model, DSS causes disruption of the intestinal epithelial monolayer lining, leading to the entry of luminal bacteria and associated antigens into the mucosa and activation of the innate immunity .w/v) ad libitum for 7 days, and then replaced with plain water. Cannabinoid treatments, 5 mg/kg, were administered intraperitoneal, every other day, from Day 1 to Day 10 (p < 0.0001). In THCE-treated mice, the average length of the colon was 4.8 cm vs. 4.1 cm in the THC-treated mice (p < 0.0001). All treatment significantly improved the clinical condition of the mice as compared with the vehicle DSS group. The difference between the CBD and THC groups was not significant.Acute colitis was induced by adding DSS to the mice drinking water at 2% . The THCE and THC treatments were not significantly different.Next, we examined the effects of the treatments on macrophages and inflammation in the DSS model. In IBD patients, an increased number of macrophages in the inflamed mucosa initiate a rapid response to luminal microbial antigens . ImmunohCytokine levels in the blood reflect the inflammatory status in the body. It was also demonstrated that inflammatory cytokines have a crucial role in the pathogenesis of IBD, where they control multiple aspects of the inflammatory response . We, theMacrophages play key roles in innate immunity. They are specialized cells involved in the detection and destruction of bacteria and other harmful pathogens. In addition, they can present antigens to T cells and initiate inflammation. Macrophages and microglial cells express the Gi protein-coupled seven transmembrane cannabinoid receptors, CB1 and CB2 ,22,23,24Phytocannabinoids possess a wide range of immune regulatory properties, mediated by the endocannabinoid system . Two canPreviously, we compared the influence of THC, CBD, and cannabis extracts on lymphocyte activation in vitro and in a murine graft versus host disease (GvHD) model . In the 2•− by macrophages and other cells. NO• and O2•− generate large quantities of the toxic molecule peroxynitrite (ONOO−), an oxidant and nitrating agent which can damage a wide array of molecules in cells, including DNA and proteins [First, we examined the effect of cannabinoid treatments on NO• secretion from peritoneal macrophages, ex vivo. Macrophages produce large amounts of NO• as a defense mechanism. However, in pathological conditions, such as autoimmune diseases, there is excessive simultaneous production of NO• and Oproteins . Elevateproteins . We founproteins . CannabiAging is concurrent with a slow and constant functional deterioration of the immune system, known as immunosenescence, which is accompanied by an increase in chronic inflammatory processes, a phenomenon known as “inflammaging”. The devastating consequences of an aged immune system include impairment of the ability to cope with infections and an increased risk of developing chronic diseases such as cancer and heart disease—the two leading causes of death in old age . Indeed,The effect of THC and CBD treatments on the expression of cell surface molecules on activated peritoneal macrophages was examined. Stimulation with LPS, a major component of the outer membrane of Gram-negative bacteria, leads to an effective classical, proinflammatory, macrophage activation. MHC-II molecules, expressed by antigen presenting cells, are central in the initiation of cellular and humoral immune responses. Upon LPS activation, MHCII is upregulated in macrophages , and henF4/80 is widely used as a mouse macrophage antigen marker. Although the ligands for F4/80 have not been defined, evidence shows that this molecule is required for peripheral tolerance . In addiIBD is comprised of well-known gastrointestinal autoimmune inflammatory disorders such as ULcerative colitis (UC) and Crohn’s (CD). Cannabinoid receptor stimulation attenuates murine colitis, while cannabinoid receptor antagonism and cannabinoid receptor deficient models reverse these anti-inflammatory effects . PhytocaDysregulated macrophages have a key role in the pathogenesis of IBD. In IBD patients, the number of macrophages increases in the inflamed mucosa. The phenotype and functions of the macrophages in the inflamed sites differ from those in physical conditions. For instance, they express high levels of costimulatory molecules and produce high levels of IL-12 and IL-23 in vitro under the microbial stimulation [Inflammatory cytokines are produced by macrophages, but also by other cells such as dendritic cells, fibroblasts, and lymphocytes. The cannabinoid treatments all affected the levels of inflammatory cytokines in the mice blood. However, each treatment had a unique effect on the composition of cytokines: CBD treatment reduced the levels of IL6 and the more regulatory cytokine, interferon-beta; THC treatment reduced the levels of IL6 and interferon-beta better than CBD; CBDE treatment reduced the levels of TNF-alpha and had the strongest effect on the level of IL6; and THCE treatment reduced the levels of IL6, interferon-beta; and TNF-alpha. These results demonstrate, once again, the different consequences of treatment with different cannabinoid-based treatments. Interestingly, IL6, a key cytokine in IBD  was reduOverall, our results indicate both similarities and differences between the impact of CBD- and THC-based drugs. Although all the tested treatments had an anti-inflammatory effect, their specific effects  differed. These differences may influence the clinical outcome of the treatment. We were surprised to find very similar anti-inflammatory results for the two cannabis extracts, which had diverse content of THC and CBD. This could suggest that THC/CBD content may not be the best indicator for anti-inflammatory properties of a cannabis-based drug. These results highlight the need to expand the research on the interplay between cannabinoids and other phytochemicals in the cannabis extracts. A better understanding of the effects of each molecule and the synergism between these molecules on the immune response will assist physicians to provide the best possible individually targeted treatment for their patients and will allow the design of new treatments.


Leptoglossus Guérin-Méneville, 1831 species are important plant pests. Leptoglossus occidentalis (western conifer seed bug), a native of western North America, has become a highly successful worldwide invader that feeds on conifer cones and can be a significant pest in conifer seed orchards. Leptoglossus zonatus, or the large-legged bug, occurs throughout much of the Western Hemisphere and is an important pest to a wide range of crops in the southern United States. Mouthparts are necessary appendages specialised in detecting food sources and feeding. The types and quantities of various sensory organs are closely related to the feeding habits of insects. The labial sensilla occurring in the two aforementioned species were examined using field emission scanning electron microscopy, and their structural details were described for the first time. The elongated and slender, four-segmented labia have the same structure in both species. Nine types of aporous sensilla, whose function is probably mechanoreception, and three uniporous sensilla, most likely responsible for chemoreception (gustatory receptors), were found. This study increases the available detailed morphological and behavioural data for Hemiptera and contribute to improve our understanding of these pests’ feeding behaviour and sensory mechanisms.Several Leptoglossus occidentalis Heidemann, 1910 and L. zonatus  (Hemiptera: Heteroptera: Coreidae) is presented. The detailed morphology, location, and distribution of different sensilla types on mouthparts were investigated and shown in micrographs taken with a scanning electron microscope. Nine types of aporous sensilla, and three uniporous sensilla were found. The possible functions of these sensilla as well as similarities and differences between the mouthparts of Leptoglossus and those of other terrestrial Heteroptera are discussed. The tip of the labium constitutes a functional “touch and taste area”, combining the chemosensitivity of uniporous sensilla P1–P3 and the mechanoreceptivity of A8 and A9 hair-like sensilla. A set of two cone-like chemosensilla types (9 + 2) was found on each lateral lobe of the labial tip. Literature analysis showed that such a set of cone-like sensory organs on the labial tip may be common in terrestrial Heteroptera. This observation confirms that the number and arrangement of sensilla is conservative and can be important in diagnosing taxa at various levels and in phylogenetic studies based on morphology.A detailed description of the labial sensory organs of  Leptoglossus Guérin-Méneville, 1831 (Pentatomomorpha: Coreidae) [Heteroptera (or true bugs) is a monophyletic suborder within Hemiptera. It has about 45,000 species with a habitat range unmatched by other groups, including aquatic and semiaquatic habitats ,2. Many Leptoglossus is a large genus with 62 species characterised by flattened and dilated hind tibiae [Leptoglossus is neotropical in origin, with most species limited to Central and South America [d tibiae   [Leptoglossus zonatus is an important pest of a wide range of crops in the southern United States, including cotton, tomato, eggplant, almonds, pistachios, pomegranate, peach, citrus, watermelon, corn, and pecan [s, 1852)   [Neomegalotomus parvus  [Riptortus pedestris  [Coreus marginatus  [Thus far, the morphology and ultrastructure of the labial receptors of true bugs have been studied in several species within families: Nepomorpha ,25,26 Ger, 1837) , and twod, 1842)  and Ripts, 1758) .Leptoglossus species using a scanning electron microscope (SEM). The morphology, location, and distribution of different sensilla types on mouthparts are investigated, and the possible functions of these sensilla are discussed.Thus, consistent and detailed studies are needed in order to provide valuable comparative data. This study aims to provide the first detailed morphological characterisation of the labial sensory organs of two L. occidentalis collected in Poland (Upper Silesia) as well as a male and three females of L. zonatus from the neighbourhood of the town of Parlier, California (Fresno County). Specimens were picked up with tweezers and poisoned with ethyl acetate. The examined SEM preparations are preserved in the Institute of Biology, Biotechnology and Environmental Protection collections, Faculty of Natural Sciences, at the University of Silesia in Katowice (DZUS).The study is based on two males, and two females of Because of the fact that no sexual dimorphism of the labial structure was found, it was decided to present the results without distinction of the sex of specimens.The photographs  were takThe material was dissected to separate the labia from the head and cleaned in detergent using an ultrasonic cleaner. Then, the standard procedure was applied : dehydraThe lengths of the labia and their particular segments were measured using an electronic vernier by Leica application suite 4.12.0 software and presented in millimetres (mm).The identification of sensilla and the terminology used in the present study were based on the main classifications according to Altner and Prillinger  and ShieSince the structure of the labium and the sensory organs of both species showed no differences, they are discussed together.L. occidentalis and 11.56 mm in L. zonatus), and four-segmented . Except for the partially fused dorsal surface SII and SIII . The names of different morphological types of sensilla, particularly “trichoid”, “chaetic”, and “basiconic”, are used variably and interchangeably by various authors. In the current categorisation of sensilla, length is not an important criterion for classification because of the significant discrepancies in the descriptions of sensilla in the heteropteran species by different authors. Therefore, we have decided to simplify the nomenclature and include the sensilla’s features in their descriptions.A1—oval and cupola-shaped structure, not evidently protruding over the surface of the cuticle  sensilla A1 and a few scattered sensilla A2 on the basal part. Excluding the distal end, the rest of the segment is covered with sensilla A2 and A5. Ventral surface: almost glabrous; only the side edges are covered with sensilla A2 and A5.Dorsal surface: on the basal part of the segment, there are three symmetrically located pairs of sensilla A3 and a pair of sensilla A4. Additionally, in the basal part between the sensilla A2, there are scattered sensilla A5. Along the entire length of the segment, there is a single row of sensilla A2 on both sides of the labial groove. Between them, there are a few asymmetrically distributed sensilla A6. At the distal end of the segment, there are two pairs of symmetrically arranged sensilla A1. Ventral surface: almost glabrous along the midline; the sides are covered with sensilla A2 and A5.Dorsal surface: covered with numerous sensilla A2 and A5, and a few sensilla A6. Ventral surface: except for the midline, covered sparsely with short sensilla A2. The sides are covered with sensilla A2 and A5.Dorsal surface: at the segment’s basal edge, there is a symmetrically located single pair of sensilla A3. Along the entire length of the segment, there is a single row of sensilla A2 on both sides of the labial groove. Between them, in the distal half, there are a few sensilla A6. Ventral surface: covered sparsely with short sensilla A2 along its entire length. A few A6 sensilla are also present in the distal part.Labial tipThe lateral lobes of the labial tip consist of two symmetrical sensory fields. Below the lateral lobes, on the sides of the base of the apical plate, there is a pair of symmetrically located sensilla A7. Around the base of each lateral lobe, there are three sensilla A8 , a single sensillum A9  and a single sensillum P3 . At the top of the lateral lobe, there are nine sensilla P1, with two sensilla P2 in between.Apical plate (Ap)—trapezoidal plate —is a pair of cuticular protuberances covered by numerous cuticular processes of various shapes .Chemoreceptors are responsible for insects’ senses of taste and smell. The olfactory receptors detect gaseous chemicals, and the gustatory receptors detect compounds dissolved in water or even on dry surfaces ,55,58. SLeptoglossus is characterised by a unique, long base . Not all authors distinguish between P1 and P2 sensilla. However, high-quality micrographs in papers on species from various taxonomic groups allow for the analysis of sensilla. Within the Cimicomorpha, these are: Miroidea, Miridae: Lygus lineolaris [L. rugulipennis [Adelphocoris fasciaticollis [Cheilocapsus nigrescens [Stephanitis nashi [Himacerus apterus, H. mirmicoides, and Prostemma guttula [Pyrrhocoris sibiricus [Physopelta cincticollis [Cazira bhoutanica, Picromerus bidens, and P. lewisi [Leptoglossus, there are nine sensilla P1 on each lateral lobe. It is widely recognised that sensilla P1 are contact chemoreceptors and therefore have a gustatory function. This is evidenced by the presence of terminal pores and ultrastructural details [Sensilla P1 occur in many terrestrial Heteroptera and are most often referred to as sensilla basiconica ,49,51,60neolaris ,29, L. rlipennis , Adelphogrescens ; Miroideis nashi ; Naboide guttula . Within . lewisi . In all  details .Eocanthecona furcellata (Pentatomidae), sensilla basiconica type B’s  surface is perforated with numerous pores, which indicates olfactory function [Dysdercus fasciatus (Pyrrhocoridae), Peregrine [Leptoglossus and P. sibiricus [Dolycoris indicus [Sensilla P2 are also present in all of the above species , although, as already mentioned, they are not always distinguished. Similar to the P1 type, sensilla P2 are most often considered to be uniporous contact chemoreceptors. However, in the case of function . Furthereregrine  suggesteeregrine  assignederegrine . Either ibiricus  or Dolyc indicus .S. nashi [P. sibiricus, a single sensillum basiconicum IV on each lateral lobe [Sensilla P3 does not seem to have an equivalent in Heteroptera’s representatives studied so far. They look a bit like campaniform sensilla but stand out clearly above the surface and have a large, centrally located pore. In addition to the previously mentioned P1 and P2 sensilla equivalents, various sensory organs were observed on the labium tip of heteropterans. These include, e.g., oval plate sensilla with a terminal pore and placoid elongated sensilla with wall pores, which have been shown in Nabidae , multipoS. nashi . The menral lobe   were found. The tip of the labium constitutes a functional “touch and taste area”, combining the chemosensitivity of uniporous sensilla P1–P3 and the mechanoreceptivity of A8 and A9 hair-like sensilla. The set of two cone-like chemosensilla types (9 + 2) on each lateral lobe of the labial tip was found. Literature analysis showed that such cone-like sensory organs on the labial tip may be common in terrestrial Heteroptera. This observation confirms that the number and arrangement of sensilla are conservative and can be important in diagnosing taxa at various levels and in phylogenetic studies based on morphology. It should be noted that further studies, including electron transmission microscopy observation or electrophysiological and behavioural bioassays, are necessary to confirm the functions of the sensory organs proposed here.


Salmonella and Staphylococcus aureus (S. aureus) and Listeria monocytogenes were examined and the migration assays were carried out. The migration analysis results of CS1, CS2, and CS3 nanocomposite films were found as 0.305, 0.297, and 0.297 mg/dm2, respectively. The inhibition zone of CS1, CS2, and CS3 nanocomposite films were found as 1547, 386, and 1884 mm2 against Salmonella bacteria. The results show that silver nanofilms are suitable as packaging films for the production of packaging in milk and dairy products, liquid foods, and acidic foods.In this original research, biodegradable corn starch (CS) and wheat gluten (wg)-based silver nanofilms were synthesized and analyzed by using goji berry extract taurine (ta), garlic extract (GC), whey powder (wh), and montmorillonite clay nanoparticles. Antibacterial-corn-starch-based nano films were analyzed by using the methods of high-performance liquid chromatography (HPLC), Fourier Transform infrared spectroscopy (FTIR-ATR), X-ray diffraction (XRD), dynamic and mechanical (DMA) analysis, and scanning electron microscopy (SEM). In addition, the antibacterial resistances of the corn starch nano films against the bacteria  Goji berries have strong antioxidant features and a high level of beta carotene, iron, vitamins , and protein, as well as 18 amino acids and 21 minerals; moreover, they have more vitamin C than an oranger. They provide a feeling of satiety with its high fiber content of 21%. They are beneficial for treating high cholesterol and cancer since they protect retina cells and bones, etc., and they have bioactive benefits. It is also known that they provide energy, facilitate digestion, and are good for depression and anxiety disorders [The taurine is a thiol-containing free amino acid that does not participate in the structure of proteins. As an essential amino acid and pharmacologically speaking, its clinical uses are expanding. The taurine extract, which is colorless, water-soluble, and strongly acidic due to its sulfonate group, has high antioxidant and antibacterial properties, much like whey powder ,4,5.As is known, the synthetic polymers used in food packaging contain components that are very harmful to the environment and human health. Petroleum-based synthetic polymers and derivatives cause serious damage to the environment due to the waste they transform. Plastic packaging materials, which are widely used in food packaging throughout the world, are a serious problem ,7. The mS. aureus, Listeria monocytogenes, and Salmonella bacteria, and migration analyses were performed. In this study, biopolymer food-packaging films, which display antimicrobial activity and contribute to the protection of nature in a short time, were synthesized. Initially, montmorillonite (Mt) clays were modified with taurine (ta) extracts obtained from goji berries dried at different temperatures, and the organoclays were synthesized. The resulting organoclays interacted with antibacterial garlic extract and whey powder, and corn starch and wheat gluten biopolymers. The antibacterial biopolymer packaging silver nanofilms, which have high antibacterial resistance, were synthesized. Synthesized silver nanofilms were analyzed by HPLC, FTIR, SEM, and DMA. The well diffusion method was applied to synthesized biofilms for antibacterial analyses for 3), DPPH , methanol anhydrous (99.8%), nutrient agar, hydrochloric acid (37%), sodium borohydride, and sodium hydroxide were purchased from Sigma-Aldrich . Corn starch, goji berry fruit, wheat gluten, and whey powder were supplied by a local producer (Food Market in Istanbul/Turkey). Na+-montmorillonite was provided by Southern Clay Products Inc. . The physical and chemical properties of sodium montmorillonite had the following the chemical formulas: 0.33 2 Si4O10 (OH)2 6H2O, density: kg/dm3 2.860, specific surface area: 0.750 m2/kg, and CEC: 920 meq/kg; their composition was wt. %: 1.40 Na, 2.44 Ca, 9.99 Al, 8.88 Mg, 20.7 Si, 35.53 O, and 0.37 H.Garlic, silver nitrate   with an accelerating beam at a voltage of 40 kV. The organic groups on the silver nanofilms were determined by FTIR-ATR. The mechanical properties of the cornstarch silver nanofilms were analyzed by using TA Instruments’ dynamic mechanical analyzer (DMA), Model 2980, New Castle, DE, USA).Listeria monocytogenes, S. aureus, and Salmonella were used for the antibacterial analysis. The analysis procedure was applied according to the literature [The bacteria terature . The nutMigration analyses were performed by preparing 50% ethanol, 10% ethanol, and 3% acetic acid solutions.50% ethanol; simulant imitating milk and milk products,10% ethanol; simulant imitating liquid foods,3% acetic acid; simulant imitating acidic foods.The measurement method of the migration analysis has been carried out according to the literature , and TGKFor statistical analysis, the Minitab 16 software was used. ANOVA and Tukey’s test at a confidence level of 95% were used .−1 assigned to carbon–carbon and carbon–nitrogen stretching vibrations in taurine extract. The band of O-H represents water adsorption on the montmorillonite at 3627 cm−1. The stretching region of C-H is related to the modified organoclay molecules in the region of 2850–2928 cm−1. These bands are based on the C-H antisymmetric and symmetric stretching bonds. The amin peaks shown in the spectrum of the wheat gluten amide I have been defined as mainly arising from amide carbonyl stretching, a combination of amide NH bending and CH stretching that can be used to characterize the protein secondary structures. A peak at 1650 cm−1 is associated with α-helical and a random structure, and the shoulder at ≈1668 cm−1 is associated with β-turns and could also be related to glutamine side chains. The amide I region of the hydrated gluten protein broad shoulders was observed at 1654–1650 cm−1, indicating α-helix conformation [The FTIR-ATR spectra of orgaoclays showed peaks at 1668 cmormation ,22,23,24−1 may be interpreted to the stretching vibrations of C=C and C=O, respectively, which exist in the FTIR-ATR spectra of taurine and garlic-loaded polymer [−1 are higher than the spectrum of pure starch proves that the modification made by taurine and garlic extracts was successful. Moreover, it can be assumed that the interactions resulting from the modification originated from aliphatic C-H tensile bands [Moreover, the peaks at wave numbers around 1300 and 1500 cm polymer ,15,16. Tle bands ,20,21,22−1 showed C-O stretching vibrations in the C-O-C groups. The broad band that appeared at 3272 cm−1 represented hydroxyl groups bonded with hydrogen and N-H stretching bands. The FTIR-ATR spectra of the silver nanofilms are shown in −1 in −1 also represented the C-O stretching that overlapped with the N-H stretching in the same region (amide II).In the spectrum of the wheat gluten b, the peListeria Monocytogenes, Salmonella, and S. aureus. As is known, the high antimicrobial resistance of nano films to prolonging the shelf life of foods is an effective parameter in packaging applications. CS1 and CS3 silver nanofilms showed maximum resistance to Salmonella, with an area of 1547.5 and 1884.3 mm2, respectively. In addition, it may assumed that the taurine and garlic extract showed higher antibacterial resistance, especially against Salmonella bacteria. According to the results of the antibacterial analysis, the whey powder and goji berry extract taurine dried at 60 °C created larger inhibition zones, whereas the goji berry extract taurine dried at 40 °C exhibited low antibacterial performance [formance .Salmonella bacteria are found in non-typhoidal poultry, eggs, raw meat, and products made from raw eggs. Moreover, the milk and dairy products, egg products , meat and meat products, fishery products, live bivalve mollusks, live sea urchins, live tunicates, broth tablets and powders, soups in dry form, seasonings, cream, and other foods in powder and tablet form such as whipped cream and sauces can be given as examples of food types with a high risk of carrying Salmonella bacteria. For this reason, packaging syntheses that are resistant to Salmonella bacteria, which have such a high risk factor, are of great importance. 1 (c and d), CS2 (e and f), and CS3 (g and h) silver nanofilms, which formed a multilayered structure. These multi-layered structures also resulted in a rough morphology. The surface morphology of the samples was affected by the presence of taurine and garlic extract [ extract . The add extract . The str2 of packaging material per 1 kg in accordance with the European Regulation 10/2011 , according to TGK 2013/34 Article 8 in Turkey, the migration limit concentration of materials for foods is 10 mg/dm2 [1, CS2, and CS3 nanocomposite films were suitable to be packaging films for packaging production in milk and milk products, liquid foods, and acidic foods. It can be seen that all of the migration analysis results are quite below the standards values. If they are evaluated only according to the migration analysis results, it is possible to say that the synthesized silver nanofilms can be used in all food packaging, but the antibacterial analysis results limit this area, so it is recommend that they be used primarily in the food packaging of milk and dairy products.0 mg/dm2 ,28,29,30This means that the CS silver nanofilms were found to be packaging films appropriate for usage with all food products. The concentration values of the silver nanofilm’s migration analysis were calculated according to the method described in a previously published article . Table 4The tensile stress (a), elastic modulus (b), and toughness (c) analysis results of the CS and CS silver nanofilms are shown in 1, CS2, and CS3 silver nanofilms. According to the analysis results obtained, the tensile stress and toughness values of the CS2 and CS3 silver nanofilms showed more improvement than the CS1 silver nanofilm. The tensile stresses of CS2 and CS3 were increased by 68%, and the tensile stress of CS1 was increased by 8%, respectively. The reason for this can be said to originate from organoclay contained in the CS2 and CS3 silver nanofilms. As is known, organoclays are filling materials that support mechanical durability. In addition, even if the absence of organoclay in CS1 has lower tensile stress than CS2 and CS3, it can be said that it develops compared to the mechanical properties of pure CS biopolymer [1 showed more improvement than CS2, CS3, and pure CS. The increase in elastic modulus tends to decrease by 68% for the CS2 and CS3 silver nanofilms.opolymer . Moreove2 and CS3 silver nanofilms is a factor that reduces elasticity. The improvement of the mechanical properties of the synthesized silver nanofilms is also confirmed by scanning electron microscope images. The roughness of the surfaces is proof that their mechanical properties have improved. The presence of organoclay in the structure of the CSFoods are exposed to contact materials including cutlery and dishes, containers, processing machine, and packaging materials during all steps passed from farm to fork. Food industry has been conducting research and development activities on food packaging to increase shelf life, keep the food quality at optimum level, attract consumer interests, and reduce waste. A package material for any type of food should minimize aroma and flavor losses, constitute an excellent barrier for gas and water, provide a perfect hermetically sealed seam, as well as have a good mechanical properties. Food contact materials including food packaging are generally based on paper, metal, ceramic, aluminum, lacquers and coating, and plastic.Food packaging is used to increase shelf life, to keep food quality at optimum level, to attract consumer interest, to facilitate the sale and distribution. Foods packaging provides information to consumers on product name, brand name, net weight, manufacturer information, price, production date, as well as the nutrient values in addition to keeping food at the desired amount in a single vessel and making it easier to bring a number of units to be moved into a single cluster and use. Therefore, food industry makes expenditures on the research and development activities of food packaging systems. The degree of the final product quality and safety, and consumer expectations from the ergonomic features of the package affects the acceptance criteria of a package material. A package material for any type of foods should minimize aroma and flavor losses, constitute an excellent barrier for gas and water, provide a perfect hermetically sealed seam, as well as have a good mechanical properties and offer chemical and biological protection against contamination ,32.1, CS2, and CS3—were synthesized. As a result of the analyzes made, it was confirmed by the FTIR, SEM, HPLC, and DMA results that the modification produced successful results. According to the results of antimicrobial studies, it has been proven that garlic-plant extract provides positive resistance against bacteria. In the migration analysis results, it was confirmed that the synthesized antimicrobial biopolymer food-packaging films were made in accordance with Article 8 of TGK 2013/34. It has been determined that the pore structure of the CS1 film has less gas permeability than CS2 and CS3, thus creating a stronger gas barrier. This is because of the whey powder used in CS1 synthesis. The synthesized films will play an effective role in increasing the shelf life of the food products to be used. As a result, the migration analyses of the synthesized nanocomposite antimicrobial films show that they are suitable for use in food packaging. The successful results have shown that the synthesized films are not dangerous. In addition, it was concluded that the need for additives and chemical preservatives in foods can be reduced. In this way, the shelf life of foods is extended with natural bioactive ingredients.In this study, three different nanocomposite films—CS


TBCK gene, and to date, this disease was reported rather limitedly in number and all described cases were Caucasians.Infantile hypotonia with psychomotor retardation and characteristic facies type 3(IHPRF3)  is an autosomal recessive disorder caused by biallelic pathogenic variants of the TBCK variant c.247C > T(p.Arg83Ter). The parents were both carriers without any positive symptoms or signs. With an extremely low frequency (0.0000082) in Exome Aggregation Consortium, the variant has not been reported in any other databases or official literatures, and was diagnosed to be pathogenic according to the American College of Medical Genetics and Genomics(ACMG) standards and guidelines. Neurorehabilitation training did not work well and the long-term prognosis remained to be observed.This paper reported the clinical and genetic features of a Chinese patient with IHPRF3. The patient was a 15-month-old male with global developmental delay, profound hypotonia, and typical facial dysmorphic features including mildly coarse facial appearance, hypertelorism, tented upper lip, exaggerated Cupid’s bow, macroglossia and arched eyebrows. Magnetic Resonance Imaging (MRI) analysis of the brain revealed slightly widened bilateral ventricles and subarachnoid space. On genetic analysis, the patient was homozygous for a novel TBCK mutation, which provided a novel molecular marker for the definite diagnosis of IHPRF3 patients and for its genetic counseling and prenatal diagnosis in the affected families.This study reported the clinical and molecular features of the first non-Caucasian patient with IHPRF3 arising from a novel homozygous  TBCK (TBC1 domain containing kinase) gene, which is located on chromosome 4 with a total of 26 exons and encodes an 893-aa protein with kinase, TBC  and Rhodanese homology domains [TBCK is involved in the regulation of cell proliferation, cell growth, and actin organization by making an impact on the expression of mammalian target of rapamycin (mTOR) complex (mTORC) components at the transcriptional level. The main clinical manifestations included systemic growth retardation, special facial features, drug-resistant epilepsy, and chronic respiratory failure. Other frequently occurring phenotypes included poor language expression ability, hypotonia, visual impairment, hearing impairment and bone metabolism disorders [Infantile hypotonia with psychomotor retardation and characteristic facies type 3 (IHPRF3) is an infantile encephalopathy syndrome caused by biallelic pathogenic mutations in the The first case of IHPRF3 was reported in 2008 –6. HenceThe patient was a 15-month-old boy with global developmental delay noticed for 10 months. He was found to be unable to raise his head at 5 months of age when electroencephalogram was normal but brain Magnetic Resonance Imaging (MRI) . No jaundice was observed in the skin and sclera. Special facial features were observed, including mildly coarse facial, hypertelorism, tented upper lip, exaggerated Cupid’s bow, macroglossia, open mouth and arched eyebrows  analysis was conducted, and a score of 8.9 for gross motor function was got, along with the scores 23.7 for fine motor function, 26.6 for adaptive behavior, 26.6 for language ability and 26.6 for social skill, indicating global developmental delay.TBCK mutation c.247C > T(p.Arg83Ter) in the patient, which was a nonsense mutation located in exon 3. Sanger sequencing confirmed that the child was homozygous for this variant, and his healthy father and mother were both carriers  was carried out to explore the underlying genetic etiology. As a result, NGS detected a homozygous Based on the clinical and genetic findings above, the patient in this study was diagnosed with IHPRF3.Neurorehabilitation training was continued, but on a follow-up at his age of 17 months, no significant improvement was observed in the clinical manifestations such as growth retardation. The long-term prognosis remained to be observed.Figure TBCK mutation c.247C > T which caused transcription to stop at 83-aa, giving rise to a truncated protein p.Arg83Ter,which met the PVS1 criteria in the American College of Medical Genetics and Genomics (ACMG) standard and guidelines [In this study, the patient had phenotypes of severe global developmental delay, while low muscle tension and strength, special facial features and macrocephaly at birth were noticed. Although positive, the brain MRI findings was not severe or specific. On genetic analysis, the child was a homozygote of the TBCK was documented to play an important role in the control of cell proliferation, cell size, and actin-cytoskeleton dynamics by modulation of the mTOR signaling network [ network , 8. And  network . The pat network . WhetherTBCK influences the expression of mTOR components at the transcriptional level [At present, the treatment of IHPRF3 was mainly symptomatic and supportive, including regular follow-up, rehabilitation training and giving respiratory support if necessary to delay the progression of the disease. Of note, al level , and theal level , and leual level , 12. Howal level . DelayedTBCK mutation, which provided a novel molecular marker for the definite diagnosis of IHPRF3 patients, and for its genetic counselling and prenatal diagnosis in the affected families.In conclusion, this study reported the clinical and molecular features of the first non-Caucasian patient with IHPRF3 arising from a novel homozygous


In order to allow a more standardized and tailored training approach, the connection between FATmax and the individual aerobic thresholds (AerT) has been examined. Although recent findings strongly suggest that a relationship exists between these two intensities, correlation alone is not sufficient to confirm that the intensities necessarily coincide and that the error between the two measures is small. Thus, this systematic review and meta-analysis aim to examine the agreement levels between the exercise intensities matching FATmax and AerT by pooling limits of agreement in a function of three parameters: (i) the average difference, (ii) the average within-study variation, and (iii) the variation in bias across studies, and to examine the influence of clinical and methodological inter- and intra-study differences on agreement levels. This study was registered with PROSPERO (CRD42021239351) and ClinicalTrials (NCT03789045). PubMed and Google Scholar were searched for studies examining FATmax and AerT connection. Overall, 12 studies with forty-five effect sizes and a total of 774 subjects fulfilled the inclusion criteria. The ROBIS tool for risk of bias assessment was used to determine the quality of included studies. In conclusion, the overall 95% limits of agreement of the differences between FATmax and AerT exercise intensities were larger than the a priori determined acceptable agreement due to the large variance caused by clinical and methodological differences among the studies. Therefore, we recommend that future studies follow a strict standardization of data collection and analysis of FATmax- and AerT-related outcomes.Regular exercise at the intensity matching maximal fat oxidation (FAT During  highest ,3. As th highest . With th highest . max is of high interest for athletes as training at this intensity can improve lipid oxidation during exercise, which is associated with the performance of endurance-trained individuals [max has also gained attention among public health professionals and has been recommended for treating several chronic metabolic health issues [max training provides a beneficial alternative for optimizing fat oxidation during exercise, which has proven to be an effective strategy for improving body composition, insulin sensitivity, and oxidative capacity in patients with obesity, metabolic syndrome, and type 2 diabetes mellitus [max-based training [Regular exercise at FATividuals ,6. Moreoh issues ,6. In pamellitus ,8. Concrtraining ,10,11. 2) or heart rate (HR), expressed either in absolute , respectively) or relative  values [max (35–75%VO2max and 45–65%HRmax) has been revealed [The accurate prescription of individualized exercise intensities is a complex and still controversial task ,13,14. T) values . Indeed,revealed ,16. Prevrevealed ,18,19. TTwo metabolic thresholds have been traditionally identified, defining three distinct zones of energy production . In thismax and AerT in different populations [max and AerT [max and AerT seems to be influenced by the variations in the studies’ methodological approaches, fore mostly used for the estimation of both physiological biomarkers [max and AerT intensities can be used interchangeably and yield an acceptably low error at the individual level. Even though the presence of a strong association between FATmax and AerT confirms that both outcomes are correlated, knowing the error for a given individual—which is of paramount importance for aerobic exercise prescription purposes—remains unknown. For example, Rynders et al. [max and AerT despite observing a strong association between both parameters. On the contrary, Gonzales-Haro [max and AerT have been thoroughly investigated [max and AerT is still a controversial topic that requires further research. In order to increase the accuracy and effectiveness of training prescription ,16, scieulations ,25,26. Hulations ,25,26. Aand AerT . It is womarkers . Despiteomarkers , the ress et al.  reportedles-Haro  revealedstigated ,31, the max and AerT and identify relevant moderators affecting their agreement.Hence, the purpose of this systematic review and meta-analysis is to examine the agreement between the exercise intensities at FATThe present systematic review was registered in both PROSPERO  and the ClinicalTrials.gov website (NCT03789045). We used the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) checklist/flow-diagram in order to (i) structure this document, (ii) effectively explain the methodology, and (iii) systematically report the search results .max and AerT in humans. For this purpose, we used a Boolean Logic limiting the search results through different operators  in order to identify the manuscripts that reported the above-mentioned biomarkers. The following terms were combined to account for differences in the terminology [max concept was initially referred to in 2001 [A systematic search in MEDLINE (via PubMed) and Google Scholar was conducted for the identification of studies investigating exercise intensities at FATminology :  or (ii) the levels of agreement of Bland–Altman analysis. If one of the requirements failed to be reported or could not be determined from the full-text, the corresponding author was contacted and asked to provide either the missing information or the individual subjects’ raw data, allowing us to compute the data. Nevertheless, independent of its form, all studies required a clear and reproducible description of the methods used to determine both AerT and FATmax to be considered eligible. Case reports, editorials, reviews, and opinion papers were excluded. In addition, only studies that included participants with no evidence of any metabolic, pulmonary, or cardiovascular diseases  were considered acceptable. No other restrictions to participants’ characteristics were introduced other than age (18–60 years). Report criteria required studies to be written in English and to be published in a peer-reviewed journal. The grading of recommendations, assessment, development, and evaluation (GRADE) approach was applied to define the quality of a body of evidence for selected studies [In the present systematic review, only original studies  in the form of full text, abstracts, or congress presentations were included. Selected articles were classified by using the defined population, intervention, comparison, outcomes, and study design (PICOS) . Moreove studies . A tool  studies .Data extraction of the articles that met the inclusion criteria was performed in an unblinded standardized way, by four independent reviewers , using a data extraction Excel spreadsheet . Any disagreements about data extraction were resolved by consensus between the reviewers.max were assessed by using a Bland–Altman meta-analytic approach [max and AerT, the agreement analyses were performed separately for each of the measurement units identified. The primary outcome of the Bland–Altman meta-analyses was the limits of agreements (LoAs) of the differences in exercise intensities matching AerT and FATmax, and secondarily their mean bias. For each study, the mean bias  and the standard deviation (SD) of the bias were retrieved or imputed from the AerT and FATmax means, SDs, and Pearson correlation (r) or bias LoA of the selected studies. In one study [r between AerT and FATmax was estimated as the median correlation of the studies having the same measurement unit  [All statistical analyses were performed with R software (version 4.0.4)  by using metafor (version 3.8.1), clubSandwich (version 0.5.8), forester (version 0.2.0), and outliers (version 0.15) packages ,37,38,39approach . Due to ne study , due to  = 0.76) , which w2)) and the between-study variation ) were used to account for different sources of variability and to compute population LoAs as follows: δ ± 2√(σ2 + τ2) [Specifically, both the average within-study variation  and of the within-study variations in the differences (σσ2 + τ2) ,43. Addi2. Second, the study (logged) variances were modeled using separate (multilevel) random effects, providing an estimate of σ2. The standard errors of estimates of δ and σ2 were estimated using Robust Variance Estimation (RVE), while the standard error of τ2 was estimated using a formula previously provided [Estimating the LoAs and their 95% CIs required several steps. First, the study mean biases were modeled using a (multilevel) random-effects model, providing estimates of δ and τprovided . Third, provided .max  [In analyses in which there was only a single effect size from each sample, standard random-effects models were used . In analctively) . In ordectively) ,45. Addictively) . The samctively) . When a ctively) .The Grubbs test was used to evaluate the presence and effect of the outliers in the current data, and a cut-point for such outliers’ recognition was estimated ; such oumax zone  [max zone reported for the corresponding measurement unit, then AerT could be used for optimizing fat oxidation with an acceptable error. The R software code used in this study is freely available in Finally, a priori determination of a clinical and physiologically practical acceptable range of the LoA was based on the concept of FAT FATmax) , which w FATmax) . If the The systematic search retrieved a total of 72,205 papers with 18,420 identified as duplicates. After the screening of the paper’s title and language, a total of 675 manuscripts were retained. Finally, articles’ abstracts were screened and a total of 89 records were selected to be read in full as potentially eligible. An additional 77 articles were excluded after reading the full text as they did not meet the inclusion criteria. The literature search and study selection process are reported using the PRISMA statement in When the selected studies were examined for their clinical diversity, two moderators were identified: (i) sex and (ii) physical activity level, each having two different subgroups . The incmax and AerT and, consequently, agreement. Due to the high variability present, only this moderator is presented according to the effect sizes (max) was used, whereas in five (11.1%) cases, HR beats per minute (b/min) was used. For two (4.4%) effect sizes, absolute VO2max values were used (L/min). A %VO2max was used in 12 (26.7%) cases, whereas relative VO2max values (mL/kg/min) were used in 13 cases (28.9%). As for the ergometry type used, cycle ergometry was the preferred method in seven (58.3%) studies, whereas a treadmill was employed in five (41.7%). When considering the methods used to determine the AerT, four (33.3%) studies preferred the lactate method, whereas gas analysis was used in six (50%). On two (16.7%) occasions, the studies used both methods. Regarding the protocols used to assess VO2max/AerT, our results showed that all studies favored a graded exercise test (GXT) type protocol. Even though the included studies used GXT protocols ranging anywhere from 1 min to 5 min, no two studies implemented the same GXT protocol. Hence, due to high variability and to assure appropriate statistical analysis, stage lengths ≤3 min were classified as short, whereas those >3 min were considered as long. Ten (83.3%) studies preferred shorter stages while longer stages were used in only one study (8.3%). One (8.3%) study used both short and long stages. FATmax identification was performed by using visual inspection of the appropriate plots in seven (58.3%) studies, while five (41.7%) studies identified this occurrence by using a mathematical model. Lastly, ten (83.3%) studies determined FATmax during the assessment of VO2max/AerT, whereas two (16.7%) studies used an additional test.When the selected studies were examined for their methodological diversity, six moderators with relevant subgroups were identified . The prict sizes . In thremax, expressed as mL/kg/min and %VO2max, the effect of pooling study distributions can be seen in Assuming the normal distribution of differences between AerT and FAT2max, respectively  at the individual level when aiming to interchangeably use FATmax and AerT exercise intensities. In the present study, the agreement between FAT2 is expressed in mL/kg/min, the main factors (moderators) affecting the agreement between FATmax and AerT are the physical activity level , the type of ergometer used , the analytical process used to identify the AerT , and the duration of GXT exercise stages . However, the same trends were not observed in the agreement analyses between FATmax and AerT when exercise intensities are expressed in %VO2max where apparently none of the moderators have an effect on the agreement. Follow-up subgroup analyses show that, when VO2max, and a higher heterogeneity due to different individual characteristics caused by the practice of different sports types . The results obtained for the physical activity level (using mL/kg/min) moderator could be due to the fact that the subgroup including inactive individuals was characterized by studies that included participants with a similar lower fitness level , creating a narrower range of possible exercise intensities (resulting in narrower LoAs) for this subgroup. Contrarily, the fitter individuals were characterized by a wider range of exercise intensities due to a higher VO2max) moderator could be related to the exercise intensity range and the fact that it can always reach 100% of %VO2max, resulting in a greater normalization of the exercise intensity range, independently from the physical activity type. Therefore, the narrower LoAs observed for the exercise intensity expressed in mL/kg/min might be related to a lack of standardization in this particular method and its smaller scale caused by participants with lower fitness level. This could be further explained by both physiological and methodological factors. From a physiological standpoint, the fitness level and the training exercise intensity can dictate the levels of improvement of the AerT [max [max and AerT tend to be affected by similar factors  and, as shown in a recent systematic review [max should cause a shift in both parameters simultaneously when certain factors change. Consequently, the factors that affect the exercise intensity at which FATmax and AerT appear do not necessarily affect their agreement. According to our data, such a reaction was only observed in inactive subjects, whereas active individuals failed to demonstrate such a connection, leaving this open for further debate.On the other hand, the results observed for the physical activity level  [max and AerT. As previously mentioned, our results showed narrower LoAs for mL/kg/min when compared to %VO2max. It is worth noting that comparing LoAs coming from different measurement units with different scales will always lead toward a higher range if % is used. Therefore, between-measurement-unit differences should be taken with care and not directly compared. Indeed, when interpreting the main factors (subgroups) moderating the agreement , a probable cause of our observation could be due to the smaller scale. Therefore, the lack of a visible important effect of a subgrouping variable when a different measurement unit is used is presumably due to the inability of one of the measurement units to standardize the intensity in the different subgroups, which could be due to the different scales of the units and not directly due to the moderator (subgroup) effect. Another limitation of the obtained results is that the agreement for all measurement units used to prescribe aerobic exercise intensity was not assessed. As in the case of the moderators, this was due to the limited number of studies presenting agreement for certain measurement units  on the agreement, a limited number of the included studies and a large presence of methodological between-studies differences failed to allow a more detailed subgrouping, leaving some questions unanswered . Further on, only those studies performed in young and middle-aged adults were included into this meta-analysis. Previously, Tolfrey et al.  reportedi et al.  reported: 3–26%) ,62; hencmax and AerT intensities is low and seems to be primarily affected by the measurement unit used to define exercise intensity matching AerT and FATmax followed by the influence of certain moderators. Therefore, prescribing exercise based on the AerT to maximize fat oxidation should be performed cautiously because, due to the high variability, there is a substantial possibility that AerT could fall well outside the FATmax zone.Our results showed that the agreement between FAT


A urinary tract stone is a common urologic problem in Asia. In recent years, the incidence of urinary tract stones has increased but tends to be neglected by people with a lack of knowledge and low socioeconomic status. The occurrence of a bladder stone weighing more than 100 grams is unusual. We report the case of a 46-year-old Indonesian male patient who presented with complaints of lower abdominal pain, urinary pain, frequency, urgency, and hematuria. The patient also had severe anemia due to chronic hematuria. The patient then received open cystolithotomy because the ultrasonographic and X-ray findings of the kidney, ureter, and bladder revealed a giant bladder stone. This bladder stone measured 62 × 59 mm and weighed 301 grams. Gender, age, living in a tropical country, and lack of knowledge contributed to the patient’s neglect of symptoms. Based on data from the Indonesian Ministry of Health, the incidence of urinary tract stones in the country in 2002 was 37,636 new cases, with 58,959 visits. The total number of patients treated was 19,018, with 378 deaths (3). Urinary tract stones can be found in many locations, such as the kidney, ureter, urethra, and bladder.A urinary tract stone is one of the most prevalent urologic diseases in Asia. The frequency, incidence, and composition of urinary tract stones vary around the world and have altered over the previous few decades, with the prevalence ranging from 7% to 13% in North America, 5% to 9% in Europe, and 1% to 5% in Asia. Urinary tract stones are prevalent in 5%-19.1% of the population in West Asia, Southeast Asia, South Asia, and some developed countries (South Korea and Japan), but just 1%-8% of the population in East Asia and North Asia (4). This case report presents the case of a giant bladder stone in a 46-year-old male patient, who complained of abdominal pain, urinary pain, frequency, urgency, and hematuria. The patient eventually received open cystolithotomy because of the massive size of the stone.A bladder stone weighing over 100 grams is a rare occurrence We report the case of a 46-year-old Indonesian male patient, who was admitted to the Pambalah Batung General Hospital, with the chief complaints of lower abdominal pain, urinary pain, frequency, urgency, and hematuria. The following symptoms were absent: fever, nausea, vomiting, nocturia, and urinary incontinence. From the history taking, it was known that the patient had passed a small stone when urinating about 5 years ago. The patient’s daily job was a farmer, and he had graduated junior high school.On a physical examination, the patient’s vital signs were normal; abdominal palpation was within normal limits and the bladder was not palpable. An assessment of the patient’s radiological and blood laboratory examinations was also carried out.An X-ray of the kidney, ureter, and bladder (KUB) revealed the giant bladder stone shadow , and ultPreoperative blood laboratory examinations showed severe anemia, with a hemoglobin value of 5 g/dL . The patAs the stone size was very large, open cystolithotomy was performed to remove the stone from the bladder. The operation was performed under general anesthesia and minimal blood loss was reported.Proteus mirabilis bacteria . Bladder stones account for 5% of all occurrences of urinary tract stones and are caused by bladder blockage, neurogenic urinary dysfunction, urinary tract infections, or foreign substances (7). Adults with bladder stones are common in non-endemic areas. Men are more likely to be affected than women (8). Men are more likely to consume excessive alcohol and coffee and more meat than women. The testosterone hormone in men also increases the likelihood of stone formation, whereas estrogen in women seems to inhibit it (1). Otherwise, in endemic areas, children are also at high risk of developing bladder stones even without major anatomic abnormalities (9). The main influencing factor is the climate ; lifestyle, lack of fluid intake, lack of education, and low socioeconomic factors also contribute to the formation of bladder stones (10). Bladder stones are frequent in Turkey, Iran, India, China, and Indonesia as a result of malnutrition in the early years of life; however, as social conditions improve, the incidence of bladder stones is decreasing (8). The main risk factors found in our patient was male gender, age 46 years, living in a tropical country, and lack of knowledge (he had graduated junior high school).Epidemiological studies of urinary tract stones had shown a prevalence of between 1% and 19.1% in Asia; the highest incidence was found in the age group of 30-60 years (6). From history taking, we discovered the symptoms of lower urinary tract infection, lower abdominal pain, and gross hematuria in our patient. Bladder stones are mostly associated with kidney or ureteral stones and rarely occur without associated upper urinary tract stones, as was found in our patient (5). Primary bladder stones, which are made up of ammonium urate and calcium oxalate, are relatively frequent in Asia (8). After extraction, laboratory findings showed that our patient’s bladder stone did contain calcium oxalate.Bladder stones are characterized by recurrent urinary tract infections, hematuria, and urinary retention (7). Endourologic equipment with a reliable energy supply has just been introduced. Endourologic therapy for bladder stones might be advised as a safe and effective noninvasive treatment option (12). From January 2011 to April 2015, Deswanto et al. examined data from the medical records of 92 patients with bladder stones at Cipto Hospital Mangunkusumo. In these investigations, 49 patients with bladder stones measuring 2.5 ± 2.0 cm received extracorporeal shock wave lithotripsy (ESWL), with 46 patients (93.9%) remaining stone-free, and 33 patients with bladder stones measuring 4.2 ± 2.8 cm underwent intracorporeal lithotripsy, with a 97% stone-free rate. Sectio alta was only performed in 10 of 92 patients, with a 100% stone-free rate (11). According to the findings of Cicione et al., the stone-free percentage of ESWL in bladder stones ranged from 72% to 99%. In most cases, no anesthetic is required for the procedure. A urethral catheter is used to fill and empty the bladder, making stone localization and fragment clearance easier. However, around 17% of patients require cystoscopy to remove the pieces, and 10%-25% of patients require repeat therapy (12).The preferred method for diagnosis is cystoscopy, but an X-ray or ultrasound is sometimes sufficient (7). USG and KUB X-ray assessments were performed on our patient and revealed a giant bladder stone; based on these considerations, cystolithotomy was finally performed. The size of the stone is the most significant point to consider when selecting a treatment method. Endourologic equipment with an effective energy source has recently been created, although it is difficult to take calculi larger than 2 cm. The patient’s general condition, signs, and symptoms improved after the surgery (12).Cystolithotomy is an acceptable treatment for big bladder stones because of their size A giant bladder stone with minimal symptoms is a rare case and should be treated with cystolithotomy. A giant bladder stone, measuring 62 × 59 mm and weighing 301 grams, was extracted intact from the patient’s bladder. In this case, the patient developed severe anemia as a result of prolonged hematuria, which was neglected by the patient for about 5 years. Laboratory findings revealed that the stone was calcium oxalate, which is the most common compound found in giant bladder stones. Gender, age, living in a tropical country, and lack of knowledge contributed to the patient’s neglect of symptoms.NoneThe authors thank PubMed, the DOAJ, and Cochrane for the accessibility and wide publication of the journals that were collected and analyzed in this study.All authors contributed to the study conception and design, performed data collection, made substantial contributions to the analyses and interpretation of the data, and wrote this manuscript. All authors read and approved the final manuscript.The patient had agreed and signed informed consent regarding publishing the case in an academic journal without exposing his identity.Not applicable