BACKGROUND:In vitro construction of tissue-engineered intestinal models plays an important role in intestinal regeneration and intestinal disease research.The interaction of intestinal nervous system and intestinal epithelial barrier to maintain body homeostasis is a hot topic in the bionic construction of tissue-engineered intestinal tract.OBJECTIVE:To construct a bionic model that can mimic the enteric nervous system in vivo.METHODS:Using fibroin protein with villus structure as scaffold,human induced neural stem cells solidified with collagen were added to intestinal epithelial cells(Caco-2 and HT29-MTX-E12)for 3-day culture to construct a co-culture system of intestinal epithelial cells and nerve cells(co-culture group).Human induced neural stem cells or intestinal epithelial cells cultured alone that were inoculated with fibroin scaffolds were set as controls.Cell morphology was observed by scanning electron microscopy and hematoxylin-eosin staining.Cell activity was detected by Live/Dead cell staining.Human induced neural stem cell differentiation was detected by β-microtubulin immunofluorescence staining.Intestinal epithelial histological properties and barrier function were detected by microvillin,sucrase-isomaltase,tight junction protein 1,E-calmodulin,and mucin-2 immunofluorescence staining.The function of mucus secretion from intestinal epithelial cells was detected by Alcian blue staining.Alkaline phosphatase staining was performed to detect differentiation of intestinal epithelial cells,at the same time,sucrase-isomaltase,tight junction protein 1,and alkaline phosphatase mRNAs were detected by RT-qRCR.RESULTS AND CONCLUSION:The neuralized intestinal mucosal co-culture model with villi structure was successfully constructed,and neural stem cells and intestinal epithelial cells on the fibroin scaffold showed good cellular activities.After neuralization,the activity of alkaline phosphatase and sucrase-isomaltase in intestinal epithelial cells was enhanced,while the expression level of tight junction protein 1 was up-regulated.To conclude,the neuralized bionic intestinal epithelial model is beneficial to the maturation of intestinal mucosal epithelial cells and the formation of barrier function.

Objective: To understand the impact of childhood trauma on psychological distress among upper grade elemetary school students, and to explore the mediating role of leisure activities in the relationship, so as to provide a basis for developing mental health intervention strategies. Methods: From August to November 2024, a combination of convenience sampling and stratified cluster random sampling was employed to recruit 1 373 fourth to sixth grade students from four primary schools in Harbin. The Childhood Trauma Questionnaire(CTQ), a self designed leisure activity scale (including active and passive leisure activities), and the Kessler Psychological Distress Scale (K10) were used to assess childhood trauma experiences, leisure activities, and levels of psychological distress. Spearman correlation analysis and linear regression analysis were conducted to explore the relationships among childhood trauma, leisure types, leisure time, and psychological distress. Based on the mediation analysis framework proposed by Hayes (Model 4), the mediating role of leisure types in the relationship between childhood trauma and psychological distress was examined. Results: Totally 19.1% of the upper elemetary school students exhibited psychological distress, while 30.2% had experienced childhood trauma. During school days, 64.6% of the students were reported of having leisure time concentrated between 1 and 5 hours per day, whereas 67.4% reported leisure time exceeding 5 hours per day on weekends. After controlling for potential demographic confounders such as gender, grade, ethnicity, household registration, being an only child, parents educational level, co residence, and whether parents are first time married,linear regression analysis showed that childhood trauma experience had positive predictive effect on psychological distress in upper primary school students( β =0.20, P <0.01). Leisure time showed no statistically significant association with psychological distress, both on school days ( β =-0.58 to -0.56) and weekends ( β =0.26- 0.98 )(all P >0.05). Active leisure activities were negatively associated with psychological distress ( β =-0.20), while passive leisure activities were positively associated with psychological distress ( β =0.29)(both P <0.01). Leisure type partially mediated the relationship between childhood trauma and psychological distress, accounting for 11.7% of the indirect effect. Conclusion Childhood trauma experiences positively predict psychological distress in upper elementary school students, and affect psychological distress through active leisure and passive leisure.

BACKGROUND:In vitro construction of tissue-engineered intestinal models plays an important role in intestinal regeneration and intestinal disease research.The interaction of intestinal nervous system and intestinal epithelial barrier to maintain body homeostasis is a hot topic in the bionic construction of tissue-engineered intestinal tract.OBJECTIVE:To construct a bionic model that can mimic the enteric nervous system in vivo.METHODS:Using fibroin protein with villus structure as scaffold,human induced neural stem cells solidified with collagen were added to intestinal epithelial cells(Caco-2 and HT29-MTX-E12)for 3-day culture to construct a co-culture system of intestinal epithelial cells and nerve cells(co-culture group).Human induced neural stem cells or intestinal epithelial cells cultured alone that were inoculated with fibroin scaffolds were set as controls.Cell morphology was observed by scanning electron microscopy and hematoxylin-eosin staining.Cell activity was detected by Live/Dead cell staining.Human induced neural stem cell differentiation was detected by β-microtubulin immunofluorescence staining.Intestinal epithelial histological properties and barrier function were detected by microvillin,sucrase-isomaltase,tight junction protein 1,E-calmodulin,and mucin-2 immunofluorescence staining.The function of mucus secretion from intestinal epithelial cells was detected by Alcian blue staining.Alkaline phosphatase staining was performed to detect differentiation of intestinal epithelial cells,at the same time,sucrase-isomaltase,tight junction protein 1,and alkaline phosphatase mRNAs were detected by RT-qRCR.RESULTS AND CONCLUSION:The neuralized intestinal mucosal co-culture model with villi structure was successfully constructed,and neural stem cells and intestinal epithelial cells on the fibroin scaffold showed good cellular activities.After neuralization,the activity of alkaline phosphatase and sucrase-isomaltase in intestinal epithelial cells was enhanced,while the expression level of tight junction protein 1 was up-regulated.To conclude,the neuralized bionic intestinal epithelial model is beneficial to the maturation of intestinal mucosal epithelial cells and the formation of barrier function.

Objective To investigate the expression and clinical significance of miR-483-3p in patients with gesta-tional diabetes mellitus(GDM).Methods A total of 100 GDM patients with 24-32 weeks of gestation who under-went routine obstetric examination in Anqiu People's Hospital were selected as the normal glucose tolerance(NGT)group,and a total of 98 healthy pregnant women of the same age of 24-32 gestational weeks were selected as the NGT group.The expression level of miR-483-3p was detected by RT-qPCR,the diagnostic value of miR-483-3p in GDM was analyzed by receiver operating characteristic(ROC)curve,the correlation between the expression levels of miR-483-3p and clinical indicators in the GDM group was analyzed by chi-square test,and the risk factors of GDM were analyzed by Logistic regression.Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-483-3p and autophagy-related protein 7(ATG7).CCK8,flow cytometry and Transwell assays were used to detect the proliferation,apoptosis,and migration and invasion ability of cells,respectively.Results The expression level of miR-483-3p in the serum of patients in the GDM group was significantly higher than that in the NGT group.The expression of miR-483-3p had diagnostic value for GDM.Pre-pregnancy BMI,FBG,FINS,HOMA-IR and TG were significantly correlated with serum miR-483-3p expression.In addition,BMI and TG before pregnancy were risk factors leading to GDM.miR-483-3p regulated HG-treated HTR-8/SVneo cell proliferation,apoptosis,migration and invasion by targeting at ATG7.Conclusions miR-483-3p is involved in the disease pro-gression of GDM and is a potential biomarker of GDM diagnosis.

Objective:To explore the dynamic change trajectory and influencing factors of postoperative fatigue (POF) in patients with early digestive tract cancer undergoing endoscopic submucosal dissection (ESD), so as to provide theoretical basis for individualized management of such patients.Methods:Using cross-sectional survey and convenient sampling method, the patients who underwent ESD for early cancer of digestive tract in the endoscopic center of the Second Affiliated Hospital of Zhejiang University School of Medicine from January to June 2024 were selected as the research objects. The questionnaires were conducted at 30 min, 24 h, 3 days, 5 days, 7 days after ESD with Christensen's postoperative fatigue score. Spearman test was used to analyze the correlation between pain, insomnia and POF. The latent variable growth model was used to identify the potential categories of POF trajectory, and the influencing factors were analyzed by Logistic regression.Results:A total of 232 patients were finally induded, with ages of 19-94(59.53 ± 13.29) years, including 120 males and 112 femalss. The POF level of patients with early cancer of digestive tract showed a downward trend one week after operation. Three postoperative fatigue trajectories were fitted, 9.05% in moderate or severe fatigue low-speed smooth descent group (C1 group); 32.76% in moderate fatigue first decreased quickly and then slowly group (C2 group) and 58.19% in mild fatigue continuous decline group (C3 group). The result of Logistic regression analysis showed that, compared with C1 group, people without religious beliefs were morelikely to enter C2 group [ β = 1.572, OR = 4.818(1.033 - 22.465), P<0.05]; compared with C3 group, patients with high pain level and severe insomnia degree were likely to enter C1 [ β = 2.621, 0.663, OR were 13.754(2.692 - 70.283) and 1.942(1.429 - 2.638), both P<0.05] and C2 [ β = 2.010, 0.491, OR were 7.464(1.890 - 29.482) and 1.634(1.348 - 1.982), both P<0.05] group. Conclusions:There were three potential types of POF in patients with early digestive tract cancer ESD. Medical staff should pay attention to patients with severe insomnia, intense pain and religious beliefs, and give staged fatigue assessment and individualized intervention, so as to reduce the level of POF and promote rapid recovery.

Objective To investigate the potential value of crocetin(CRO)in the treatment of diabetic retinopathy(DR)and its effect on signal transducer and activator of transcription 3(STAT3)activity.Methods(1)In vitro experi-ment:RPE cells were divided into C group,HG group,HG+5/10/20CRO groups and HG+20CRO+Colivelin TFA(C-TFA)group.Cells in the C group were cultured with normal glucose medium(5 mmol·L-1).Cells in the HG+5/10/20CRO groups were cultured in high glucose medium(25 mmol·L-1)with 5,10 and 20 μmol·L-1 CRO added.Cells in the HG+20CRO+C-TFA group were cultured in high glucose medium with 20 μmol·L-1 CRO and 0.5 μmol·L-1 C-TFA added.The cells in each group were cultured for 48 h.Cell viability,proliferation and apoptosis were detected using MTT,EdU staining,and TUNEL,respectively.Meanwhile,the levels of oxidative stress and inflammatory factors were detected by ELISA,and the activation level of STAT3 was detected by Western blot.(2)In vivo experiment:Rats were divided into 6 groups:NC group,DR group,20/40/80CRO groups and 80CRO+C-TFA group.NC group was normal control rats,and the other groups were STZ-induced DR model rats.NC group and DR group were gavaged with 5 g·L-1 CMC-Na.20/40/80CRO groups were gavaged with 20,40 and 80 mg·kg-1 of CRO.80CRO+C-TFA group was gavaged with 80 mg·kg-1 of CRO,and 1 mg·kg-1 of C-TFA was injected intraperitoneally.The treatment cycle was 4 weeks.HE staining and TUNEL staining were employed to examine retinal morphology and apoptosis.The concentrations of oxidative stress markers and inflammatory factors in the retina were measured using ELISA,while the activation level of STAT3 in the retina was as-sessed through Western blot analysis.Results(1)In vitro experiments:Compared with the HG group,the HG+5/10/20CRO groups exhibited increased relative cell viability,SOD and GPx activities,and EdU+cell ratio,but decreased TUNEL+cell ratio,MDA content,TNF-α,IL-1β and IL-6 levels,and STAT3 phosphorylation level(P＜0.05).Compared with the HG+20CRO group,the abovementioned indicators of RPE cells in the HG+20CRO+C-TFA group were reversed(all P＜0.05).(2)In vivo experiments:Compared with the DR group,the 20/40/80CRO groups exhibited increased SOD and GPx activities,but decreased proportion of retinal TUNEL+cells,retinal MDA content,TNF-α,IL-1β and IL-6 levels,and retinal STAT3 phosphorylation level(all P＜0.05).Compared with the 80CRO group,the abovementioned indicators of reti-nal cell in the 80CRO+C-TFA group were reversed(all P＜0.05).Conclusion CRO can alleviate high glucose-induced rat RPE cell injury and retinal injury of DR rats by inhibiting STAT3 activity.

Objective To investigate the potential value of crocetin(CRO)in the treatment of diabetic retinopathy(DR)and its effect on signal transducer and activator of transcription 3(STAT3)activity.Methods(1)In vitro experi-ment:RPE cells were divided into C group,HG group,HG+5/10/20CRO groups and HG+20CRO+Colivelin TFA(C-TFA)group.Cells in the C group were cultured with normal glucose medium(5 mmol·L-1).Cells in the HG+5/10/20CRO groups were cultured in high glucose medium(25 mmol·L-1)with 5,10 and 20 μmol·L-1 CRO added.Cells in the HG+20CRO+C-TFA group were cultured in high glucose medium with 20 μmol·L-1 CRO and 0.5 μmol·L-1 C-TFA added.The cells in each group were cultured for 48 h.Cell viability,proliferation and apoptosis were detected using MTT,EdU staining,and TUNEL,respectively.Meanwhile,the levels of oxidative stress and inflammatory factors were detected by ELISA,and the activation level of STAT3 was detected by Western blot.(2)In vivo experiment:Rats were divided into 6 groups:NC group,DR group,20/40/80CRO groups and 80CRO+C-TFA group.NC group was normal control rats,and the other groups were STZ-induced DR model rats.NC group and DR group were gavaged with 5 g·L-1 CMC-Na.20/40/80CRO groups were gavaged with 20,40 and 80 mg·kg-1 of CRO.80CRO+C-TFA group was gavaged with 80 mg·kg-1 of CRO,and 1 mg·kg-1 of C-TFA was injected intraperitoneally.The treatment cycle was 4 weeks.HE staining and TUNEL staining were employed to examine retinal morphology and apoptosis.The concentrations of oxidative stress markers and inflammatory factors in the retina were measured using ELISA,while the activation level of STAT3 in the retina was as-sessed through Western blot analysis.Results(1)In vitro experiments:Compared with the HG group,the HG+5/10/20CRO groups exhibited increased relative cell viability,SOD and GPx activities,and EdU+cell ratio,but decreased TUNEL+cell ratio,MDA content,TNF-α,IL-1β and IL-6 levels,and STAT3 phosphorylation level(P＜0.05).Compared with the HG+20CRO group,the abovementioned indicators of RPE cells in the HG+20CRO+C-TFA group were reversed(all P＜0.05).(2)In vivo experiments:Compared with the DR group,the 20/40/80CRO groups exhibited increased SOD and GPx activities,but decreased proportion of retinal TUNEL+cells,retinal MDA content,TNF-α,IL-1β and IL-6 levels,and retinal STAT3 phosphorylation level(all P＜0.05).Compared with the 80CRO group,the abovementioned indicators of reti-nal cell in the 80CRO+C-TFA group were reversed(all P＜0.05).Conclusion CRO can alleviate high glucose-induced rat RPE cell injury and retinal injury of DR rats by inhibiting STAT3 activity.

Objective:To retrospectively analyze the current status of consultation, clinical characteristics, and treatment status of patients with IgG4-related disease (IgG4-RD) in order to provide assistance and a basis for early and standardized diagnosis and treatment.Methods:IgG4-RD cases admitted to our hospital from June 2015 to October 2023 were collected. The details of patients' basic information, initial symptoms, department visits, laboratory and imaging findings, histopathological examination results, and treatment plans were recorded. A statistical descriptive analysis was performed on the data.Results:A total of 105 patients with IgG4-RD were included, with a median age of 59.0 (18.0, 78.0) years. The main departments visited were clinical immunology and gastroenterology (83.8%, 88/105). The median diagnostic duration was eight months, with a maximum of 300 months, and 33.3% (35/105) of patients needed over one year for diagnosis. 92 cases underwent histopathological examinations and IgG4 staining, with a total positivity rate of 87.0% (80/92). Among these, sixteen cases underwent pathological examination after surgery, with a positivity rate of 100%; the remaining 76 cases out of 92 underwent liver biopsy, with a positivity rate of 76.1%. Out of these, there were 22 cases from the pancreas, 21 from the submaxillary gland, nine from the labial gland, and seven each from the duodenal papilla and liver, with positivity rates of 81.8%, 81.0%, 55.6%, 85.7%, and 85.7%, respectively. Eleven cases (10.5%) with normal serum IgG4 were diagnosed based on multi-organ involvement and pathological results. 94 cases (89.5%) had elevated IgG4, with a predominance of＞2.70 g/L. The median follow-up period for the 87 cases was 14 months. Two cases had poor response, twelve patients relapsed, five cases relapsed without combined drug treatment after surgery, five cases relapsed due to drug withdrawal, and two cases relapsed while tapering off steroids.Conclusions:As a multisystem disease, IgG4-RD still faces the difficulties of time-consuming diagnosis and inappropriate treatment. Therefore, it is necessary to rely on a multidisciplinary collaboration model to improve the awareness level and promote the early and standardized diagnosis and treatment of patients with IgG4-RD.

Objective:To investigate the effect of thrombocytopenia and coagulation dysfunction on bleeding complications in patients undergoing percutaneous liver biopsy.Methods:The clinical, laboratory, and demographic data of patients undergoing percutaneous liver biopsy at the First Affiliated Hospital of Air Force Medical University from January 2005 to January 2024 were retrospectively analyzed. The incidence of bleeding was recorded. Univariate and multivariate logistic regression analyses were used to assess the effects of thrombocytopenia and coagulation dysfunction on the risk of postoperative bleeding. Furthermore, we assessed the bleeding risk in patients with autoimmune hepatitis.Results:A total of 2 885 liver perforations were performed in 2 364 patients, 98.4% of whom had an autoimmune liver disease. There were 27 cases of postoperative bleeding (0.9%). The univariate logistic regression analysis showed that platelet count (PLT)( P<0.05, OR=0.975), coagulation dysfunction (international normalized ratio; INR)( P<0.05, OR=6.954), and cirrhosis ( P<0.05, OR=3.857) were associated with bleeding. The multivariate logistic regression analysis showed that PLT was an independent risk factor for bleeding ( P<0.05, OR=0.975). PLT scores of 40×10 9/L and 65×10 9/L can classify the bleeding risk of patients with thrombocytopenia into high, medium, and low risk. There was no difference in the risk of bleeding between the 40×10 9/L0.05, χ2=0.10). The risk of bleeding in the 31×10 9/L≤PLT≤40×10 9/L group was higher than that in the 40×10 9/L0.05, χ2=0.10). Furthermore, the bleeding cases were considered mild as the bleeding stopped after pressure hemostasis. Conclusions:PLT is an independent risk factor for bleeding after hepatocentesis. When the PLT is >40×10 9/L, hepatocentesis can still be performed safely with appropriate management measures.