OBJECTIVE: To prepare decellularized nerve grafts from alpha-1, 3-galactosyltransferase (GGTA1) gene-edited pigs and explore their biocompatibility for xenotransplantation. METHODS: The sciatic nerves from wild-type pigs and GGTA1 gene-edited pigs were obtained and underwent decellularization. The alpha-galactosidase (α-gal) content in the sciatic nerves of GGTA1 gene-edited pigs was detected by using IB4 fluorescence staining and ELISA method to verify the knockout status of the GGTA1 gene, and using human sciatic nerve as a control. HE staining and scanning electron microscopy observation were used to observe the structure of the nerve samples. Immunofluorescence staining and DNA content determination were used to evaluate the degree of decellularization of the nerve samples. Fourteen nude mice were taken, and subcutaneous capsules were prepared on both sides of the spine. Decellularized nerve samples of wild-type pigs ( n=7) and GGTA1 gene-edited pigs ( n=7) were randomly implanted in the subcutaneous capsules. Blood was drawn at 1, 3, 5, and 7 days after implantation to detect neutrophil counting. RESULTS: IB4 fluorescence staining and ELISA detection showed that GGTA1 gene was successfully knocked out in the nerves of GGTA1 gene-edited pigs. HE staining showed that the structure of the decellularized nerve from GGTA1 gene-edited pigs was well preserved; the nerve basement membrane tube structure was visible under scanning electron microscopy; no cell nuclei was observed, and the extracellular matrix components was retained in the nerve grafts by immunofluorescence staining; and the DNA content was significantly reduced when compared with the normal nerves ( P<0.05). In vivo experiments showed that the number of neutrophils in the two groups were similar at 1, 3, and 7 days after implantation, with no significant difference ( P>0.05); only at 5 days, the number of neutrophils was significantly lower in the GGTA1 gene-edited pigs than in the wild-type pigs ( P<0.05). CONCLUSION The decellularized nerve grafts from GGTA1 gene-edited pigs have well-preserved nerve structure, complete decellularization, retain the natural nerve basement membrane tube structure and components, and low immune response after xenotransplantation through in vitro experiments.
Animals ; Transplantation, Heterologous ; Galactosyltransferases/genetics* ; Sciatic Nerve/immunology* ; Swine ; Tissue Engineering/methods* ; Humans ; Graft Rejection/prevention & control* ; Gene Editing ; Mice ; Mice, Nude ; Heterografts/immunology* ; Animals, Genetically Modified ; Tissue Scaffolds ; Decellularized Extracellular Matrix

Objective To analyze the value of combined prediction of peripheral blood CD4+,CD3-CD16+CD56+,CD8+,CD19+for the risk of death in patients with Epstein-Barr virus-associated hemophagocytic syn-drome(EBV-HLH).Methods A total of 84 patients with EBV-HLH who were treated in Beijing Friendship Hospital Affiliated to Capital Medical University and Peking University First Hospital from May 2021 to A-pril 2023 were selected and divided into a death group(17 cases)and a survival group(67 cases)according to their survival outcomes within 6 months.The general information and peripheral blood T lymphocyte subsets of the two groups were compared,multivariate Logistic regression analysis was conducted to investigate the influencing factors of death in EBV-HLH patients,and receiver operating characteristic(ROC)curve was used to evaluate the predictive value of peripheral blood T lymphocyte subsets on death in EBV-HLH patients.Results The platelet count,hemoglobin,CD4+and CD3-CD16+CD56+levels in the death group were lower than those in the survival group,while the levels of CD8+and CD19+were higher than those in the survival group(P＜0.05).Multivariate Logistic regression analysis showed that platelet count,hemoglobin,CD4+,and CD3-CD16+CD56+were all independent protective factors for the death of EBV-HLH patients,while CD8+and CD19+were independent risk factors for the death of EBV-HLH patients(P＜0.05).The ROC curve results showed that the area under the curve of combined prediction of CD4+,CD8+,CD19+,CD3 CD16+CD56+for death in EBV-HLH patients was 0.923,with sensitivity and specificity of 94.12％and 80.60％,respectively,which were significantly higher than those predicted by each individual indicator(P＜0.05).Conclusion The abnormal expression of CD4+,CD3-CD16+CD56+,CD8+,and CD19+in peripheral blood of EBV-HLH patients is associated with their survival outcomes,and the combined prediction of CD4+,CD3-CD16+CD56+,CD8+,and CD19+has high reference value for predicting patients'death.

AIM:To investigate whether melatonin can ameliorate acute myocardial infarction(AMI)by in-hibiting ferroptosis.METHODS:H9C2 cells were cultured in AnaeroPack system with low sugar and serum-free medium for 10 h to construct a cell model of AMI.Then cells were treated with melatonin and ferroptosis inducer erastin.The cell activity,reactive oxygen species(ROS),lipid peroxidation,mitochondrial membrane potential(MMP),and ferroptosis related protein expression were detected.A rat model of AMI induced by isoprenaline(ISO)injection was established to evaluate the effects of melatonin,in which the myocardial infarction size,cardiac injury,pathological changes,oxidative stress,iron ion and ferroptosis related protein expression were examined.RESULTS:Melatonin decreased the oxidative stress,lipid peroxidation and expression of ferroptosis protein in cardiomyocytes induced by hypoxia,but these effects could be impeded by the ferroptosis inducer erastin.Furthermore,in vivo experiments,we also found that melatonin im-proved the myocardial infarction size,cardiac injury,pathological changes,oxidative stress,and alleviated iron ion accu-mulation and ferroptosis.CONCLUSION:The cardioprotective effects of melatonin in AMI are associated with the inhi-bition of ferroptosis.

Objective:To investigate the clinical efficacy of open anterior myofascial repair surgery on abdominal wall incision hernia.Methods:The retrospective cohort study was conducted. The clinical data of 188 patients who underwent open anterior myofascial repair surgery on abdo-minal wall incision hernia at three medical centers, including Xiangya Hospital of Central South University et al, from December 2016 to December 2024 were collected. There were 85 males and 103 females, aged (62±12)years. Of the 188 patients, 55 cases had large incisional hernia and 133 cases had non-large incisional hernia. Observation indicators: (1) intraoperative conditions; (2) postopera-tive conditions; (3) follow-up. Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact test. Comparison of ordinal data between groups was conducted using the nonparametic test. Results:(1) Intra-operative conditions. The operation time of the 55 patients with large incisional hernia was (145±40)minutes, and the volume of intraoperative blood loss was 40.0(22.5,55.0)mL, cases with fascial defect located in the central anterior abdominal wall, the superolateral quadrant, the inferolateral quadrant were 26, 7, 22, the fascial defect area was 140(99,169)cm2, cases used with self-fixating mesh and flat mesh were 29, 26. The above indicators of the 133 patients with non-large incisional hernia were (124±34)minutes, 35.0(30.0,45.0)mL, 47, 26, 60, 25(12,40)cm2, 67, 66, respectively. There were significant differences in operation time and fascial defect area between patients with large incisional hernia and non-large incisional hernia ( t=-3.651, Z=-10.339, P<0.05), and there was no significant difference in the volume of intraoperative blood loss, defect quadrant distribution, and mesh type ( Z=-0.501, χ2=2.692, 0.086, P>0.05). (2) Postoperative conditions. Of the 55 patients with large incisional hernia, 7 cases developed postoperative seroma, including 5 cases combined with concomitant surgical-site infection, and 4 additional cases developed with surgical-site infection. Of the 133 patients with non-large incisional hernia, 15 cases developed postoperative seroma, including 3 cases combined with concomitant surgical-site infection. There was a significant difference in surgical-site infection between patients with large incisional hernia and non-large incisional hernia ( χ2=10.707, P<0.05), and there was no significant difference in postoperative seroma ( χ2=0.079, P>0.05). The duration of postoperative hospital stay was 7(6, 9)days for the 55 patients with large incisional hernia and 5(4, 6)days for the 133 patients with non-large incisional hernia, showing a significant difference between them ( Z=-6.292, P<0.05). (3) Follow-up. All 188 patients were followed up for 43(range, 29-67)months. During the follow-up, 9 patients experienced hernia recurrence, including 7 patients with large incisional hernia and 2 patients with non-large incisional hernia. For the 7 patients of large incisional hernia with hernia recurrence, 4 cases underwent reoperation and 3 cases received conservative treatment. All 2 patients of non-large incisional hernia with hernia recurrence received conservative treatment. There was no significant difference in hernia recurrence between patients with large incisional hernia and non-large incisional hernia ( χ2=8.432, P<0.05). Results of chronic pain score at postoperative 3 month showed that among 55 patients with large incisional hernia, 40 cases had mild pain, 7 cases had moderate pain, and 8 cases had severe pain. Among 133 patients with non-large incisional hernia, the above indicators were 102, 28, and 3, respectively. There was no significant difference in chronic pain score at postoperative 3 month between patients with large incisional hernia and non-large incisional hernia ( Z=-0.968, P>0.05). Conclusions:Open anterior myofascial repair surgery can be used for the treatment of abdominal wall incision hernia. Compared with non-large incisional hernia, patients with large incisional hernia have longer operation time, are more prone to surgical-site infection, have longer postoperative hospital stay, and are more likely to experience hernia recurrence.

Objective:To investigate the clinical efficacy of open anterior myofascial repair surgery on abdominal wall incision hernia.Methods:The retrospective cohort study was conducted. The clinical data of 188 patients who underwent open anterior myofascial repair surgery on abdo-minal wall incision hernia at three medical centers, including Xiangya Hospital of Central South University et al, from December 2016 to December 2024 were collected. There were 85 males and 103 females, aged (62±12)years. Of the 188 patients, 55 cases had large incisional hernia and 133 cases had non-large incisional hernia. Observation indicators: (1) intraoperative conditions; (2) postopera-tive conditions; (3) follow-up. Comparison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test or Fisher exact test. Comparison of ordinal data between groups was conducted using the nonparametic test. Results:(1) Intra-operative conditions. The operation time of the 55 patients with large incisional hernia was (145±40)minutes, and the volume of intraoperative blood loss was 40.0(22.5,55.0)mL, cases with fascial defect located in the central anterior abdominal wall, the superolateral quadrant, the inferolateral quadrant were 26, 7, 22, the fascial defect area was 140(99,169)cm2, cases used with self-fixating mesh and flat mesh were 29, 26. The above indicators of the 133 patients with non-large incisional hernia were (124±34)minutes, 35.0(30.0,45.0)mL, 47, 26, 60, 25(12,40)cm2, 67, 66, respectively. There were significant differences in operation time and fascial defect area between patients with large incisional hernia and non-large incisional hernia ( t=-3.651, Z=-10.339, P<0.05), and there was no significant difference in the volume of intraoperative blood loss, defect quadrant distribution, and mesh type ( Z=-0.501, χ2=2.692, 0.086, P>0.05). (2) Postoperative conditions. Of the 55 patients with large incisional hernia, 7 cases developed postoperative seroma, including 5 cases combined with concomitant surgical-site infection, and 4 additional cases developed with surgical-site infection. Of the 133 patients with non-large incisional hernia, 15 cases developed postoperative seroma, including 3 cases combined with concomitant surgical-site infection. There was a significant difference in surgical-site infection between patients with large incisional hernia and non-large incisional hernia ( χ2=10.707, P<0.05), and there was no significant difference in postoperative seroma ( χ2=0.079, P>0.05). The duration of postoperative hospital stay was 7(6, 9)days for the 55 patients with large incisional hernia and 5(4, 6)days for the 133 patients with non-large incisional hernia, showing a significant difference between them ( Z=-6.292, P<0.05). (3) Follow-up. All 188 patients were followed up for 43(range, 29-67)months. During the follow-up, 9 patients experienced hernia recurrence, including 7 patients with large incisional hernia and 2 patients with non-large incisional hernia. For the 7 patients of large incisional hernia with hernia recurrence, 4 cases underwent reoperation and 3 cases received conservative treatment. All 2 patients of non-large incisional hernia with hernia recurrence received conservative treatment. There was no significant difference in hernia recurrence between patients with large incisional hernia and non-large incisional hernia ( χ2=8.432, P<0.05). Results of chronic pain score at postoperative 3 month showed that among 55 patients with large incisional hernia, 40 cases had mild pain, 7 cases had moderate pain, and 8 cases had severe pain. Among 133 patients with non-large incisional hernia, the above indicators were 102, 28, and 3, respectively. There was no significant difference in chronic pain score at postoperative 3 month between patients with large incisional hernia and non-large incisional hernia ( Z=-0.968, P>0.05). Conclusions:Open anterior myofascial repair surgery can be used for the treatment of abdominal wall incision hernia. Compared with non-large incisional hernia, patients with large incisional hernia have longer operation time, are more prone to surgical-site infection, have longer postoperative hospital stay, and are more likely to experience hernia recurrence.

AIM:To investigate whether melatonin can ameliorate acute myocardial infarction(AMI)by in-hibiting ferroptosis.METHODS:H9C2 cells were cultured in AnaeroPack system with low sugar and serum-free medium for 10 h to construct a cell model of AMI.Then cells were treated with melatonin and ferroptosis inducer erastin.The cell activity,reactive oxygen species(ROS),lipid peroxidation,mitochondrial membrane potential(MMP),and ferroptosis related protein expression were detected.A rat model of AMI induced by isoprenaline(ISO)injection was established to evaluate the effects of melatonin,in which the myocardial infarction size,cardiac injury,pathological changes,oxidative stress,iron ion and ferroptosis related protein expression were examined.RESULTS:Melatonin decreased the oxidative stress,lipid peroxidation and expression of ferroptosis protein in cardiomyocytes induced by hypoxia,but these effects could be impeded by the ferroptosis inducer erastin.Furthermore,in vivo experiments,we also found that melatonin im-proved the myocardial infarction size,cardiac injury,pathological changes,oxidative stress,and alleviated iron ion accu-mulation and ferroptosis.CONCLUSION:The cardioprotective effects of melatonin in AMI are associated with the inhi-bition of ferroptosis.

As a chronic metabolic disease,type 2 diabetes poses a significant threat to human health with increasing incidence.An increasing number of studies confirm that the pathogenesis of diabetes is closely related with alterations in multiple cellular signaling pathways.Although numerous studies have reported that traditional Chinese medicine compounds prevent diabetes by modulating cell signaling pathways,asystematic review of the mechanism of action of traditional Chinese medicine compounds in modulating cell signaling pathways is still lacking.Therefore,this paper summarizes the research of type 2 diabetes prevention and treatment,which was found mainly related to the signaling pathways such as PI3K/AKT,AMPK,MAPK,NF-κB,PPAR,TGF-β.This family of signaling pathways can treat type 2 diabetes by inhibiting pancreatic islet cell apoptosis,protecting pancreatic β-cell function,ameliorating insulin resistance,inhibiting hepatic gluconeogenesis,promoting glycogen synthesis,attenuating inflammation,and resisting oxidative stress.At the same time,we analyze the problems in current research and the future development trend,in order to provide a scientific theoretical basis for the drug development and clinical application of traditional Chinese medicine compound in the prevention and treatment of diabetes.

To investigate the differences in methylation levels of cuproptosis related genes in cervical cancer and their effects on clinical prognosis.Methods:The methylation data of 310 cervical tissue specimens were acquired from public databases. The UALCAN database was used to analyze the methylation level differences of 12 cuproptosis-related genes and study their level in different stages or grades of cervical cancer. Genes with statistically significant differences were selected for prognosis analysis using the EWAS datahub. Finally, gene-enrichment analysis, pathway analysis, immune infiltration analysis, the mutation rate and tumor mutation burden (TMB) of the genes in cervical cancer were analyzed using the cBioportal database. Two independent samples rank-sum test was used for differences in methylation levels and immune cell infiltration; comparative analyses of overall survival were performed using KM survival curves and Log-rank two-sided tests. TMB analyses were performed using the Wilcoxon Test for statistical analyses; Pearson correlation analysis was used for assessment in GSEA and pathway analyses.Results:The methylationβvalue of Cyclin Dependent Kinase Inhibitor 2A (CDKN2A gene) in the cervical cancer tissues of patients was 0.075 which was significantly higher than the methylationβvalue of 0.049 in normal human tissues ( P=0.008). Dihydrolipoamide S-Acetyltransferase (DLAT gene) methylation with a β value of 0.102 was significantly higher than normal human tissue methylation with a β value of 0.08 ( P=0.002), and the methylation level β value of Lipoyltransferase 1 (LIPT1 gene) in cervical cancer tissues was 0.06,which was significantly lower than normal human tissue methylation value of 0.092 ( P=0.009). Patients with CDKN2A gene methylation levels≥0.199 had an overall survival of 14.75 years, which was lower than that of patients with methylation levels<0.199 (17.56 years) ( P=0.034).The results of gene enrichment analysis indicated that it mainly involves biological processes such as the response to type I interferon and DNA replication. The expression of CDKN2A gene is positively correlated with the number of neutrophils and dendritic cells in the tumor microenvironment( P<0.05), and negatively correlated with the number ofmacrophages( P<0.05). TMB was higher in the group of variants of the CDKN2A gene than in the group of non-variants ( P=0.019). Conclusion:CDKN2A methylation is a potential biomarker for predicting the prognosis of cervical cancer.

Objective:To prospectively follow up the patients with ileocecal inflammatory lesions, to explore the characteristics of Crohn′s disease(CD) at early stage, and to provide references for early diagnosis of CD.Methods:From January 2013 to December 2018, at Department of Gastroenterology, The Seventh Medical Center of PLA General Hospital, 232 patients with unexplained ileocecal inflammatory lesions under colonoscopy examination were enrolled, which were followed up for more than one year. Chi-square test and Fisher exact probability text were used to compare the patients with early CD, with non-specific enteritis and intestinal tuberculosis in abdominal symptoms (abdominal pain, diarrhea, abdominal distension, constipation, hematochezia, changes in bowel habits), accompanying symptoms (oral ulcer, arthralgia), the proportion of patients with elevated erythrocyte sedimentation rate (ESR) or elevated C-reactive protein (CRP) level, serum antineutrophilic cytoplasmic antibody (ANCA), anti-saccharomyces cerevisiae antibody (ASCA), tuberculosis infection of T cells spot test, positive rate of fecal occult blood, lesion size, morphology, involvement site under endoscopy and histopathological results. Multivariate binary logistic regression was used to analyze the related factors of early CD.Results:Among 232 patients, 155 were males and 77 were females, and the age of first diagnosis was (43.9±13.8) years old. The follow-up period (range) was 27 months (12 to 79 months). Twenty-nine cases (12.5%) were diagnosed as early CD, 45 cases (19.4%) were intestinal tuberculosis, 105 cases (45.3%) were non-specific enteritis, and 53 cases (22.8%) as undetermined. All of 29 patients with early CD had abdominal symptoms, which accounted for 16.9% (29/172) of 172 patients with ileoceccal inflammatory lesion as well as abdominal symptoms. In early CD patients, the proportions of patients with abdominal pain, elevated CRP level and ESR level, positive rate of ASCA, positive rate of tuberculosis infection T cells and percentage of patients with thickened intestinal wall were all higher than those in patients with non-specific enteritis (62.1%, 18/29 vs. 33.3%, 35/105; 13.8%, 4/29 vs. 0; 13.8%, 4/29 vs. 1.0%, 1/105; 24.1%, 7/29 vs. 1.0%, 1/105; 20.7%, 6/29 vs. 3.8%, 4/105; 95.7%, 22/23 vs. 0), and the proportion of patients without abdominal symptoms was lower than that of patients with non-specific enteritis (0 vs. 31.4%, 33/105). And the differences were statistically significant ( χ2=6.692, Fisher exact probability text, χ2=7.162, χ2=17.826, χ2=7.497, Fisher exact probability text, and Fisher exact probability text, all P<0.05). Early CD patients were more likely to have multiple lesion sites (55.2%, 16/29), and mainly deep ulcers (55.2%, 16/29) and ulcers with a long diameter of 5 to 10 mm (39.3%, 11/28). The lesions of non-specific enteritis were mostly confined to the end of ileum (75.2%, 79/105), which were mainly superficial ulcers (41.0%, 43/105) and ulcers with a long diameter less than 5 mm (69.0%, 49/71). The proportion of patients without abdominal symptoms and the positive rate of tuberculosis infection of T cells spot test of early CD patients were both lower than those of intestinal tuberculosis group (0 vs. 15.6%, 7/45 and 20.7%, 6/29 vs. 68.9%, 31/45). The positive rate of ASCA and the proportion of patients with thickened intestinal wall were higher than those of intestinal tuberculosis group (24.1%, 7/29 vs. 0 and 95.7%, 22/23 vs. 11/19), and the differences were statistically significant (Fisher exact probability text, χ2=13.713, Fisher exact probability text and χ2=6.710, all P<0.05). The results of multivariate binary logistic regression analysis showed that abdominal pain and positive ASCA were independent risk factors for early CD (odds ratio ( OR)=2.855, 95% confidence interval ( CI) 1.014 to 8.037, P=0.047; OR=10.033, 95% CI 2.274 to 44.250, P=0.002). Conclusions:Prospective follow-up for more than one year in patients with unexplained ileocecal inflammatory lesions can effectively identify and diagnose early CD. Ileocecal inflammatory lesions with abdominal symptoms are one of the early manifestations of CD. Abdominal pain and positive serum ASCA at the initial diagnosis are independent risk factors for early diagnosis of CD.

Objective To investigate the outcomes of patients with diabetic retinopathy (DR) in Shanghai communities and the influencing factors.Methods From October 2015 to April 2016,533 type 2 diabetic patients with DR were selected by target sampling and cluster random sampling method from six community health service centers in Shanghai.Patients were followed up for 1 year.The demographic information,physical examination,laboratory tests and eye fundus exam results were documented and the DR was graded.The factors associated with the regression of DR were analyzed.Results Total 478 patients,including 280 females (58.6％) and 198 males (41.4％),were followed up for 1 year.The mean age of patients was (64±7) years and the mean disease duration was (8.85±4.20) years.The original DR lesion was remitted in 35 patients with an improvement rate of 7.3％;while the original DR lesion was aggravated in 29 patients with a progression rate of 6.1％.Ordinal logistic regression analysis revealed that age (OR=0.197,95％CI:0.056-0.699),body mass index (BMI) (OR=0.383,95％CI:0.171-0.856),glycosylated hemoglobin (HbAlc) (OR=0.287,95％CI:0.102-0.803),triglycerides (TG) (OR=0.541,95％CI:0.295-0.991),urinary albumin to creatinine ratio (ACR)(OR=0.218,95％CI:0.066-0.720) were associated with DR in type 2 diabetic patients.Conclusion The regression of DR is closely related to age,BMI,glucose,serum lipids and renal function,so it is suggested that lowering BMI,controlling glucose and serum lipids and maintaining normal kidney function are necessary for preventing the progression and promoting the improvement of DR in diabetic patients.