[摘 要] 目的：开发新型溶瘤呼肠孤病毒（Reo）递送系统，以克服中和抗体对Reo的中和作用并提升其肿瘤靶向性。方法：通过切向流过滤联合超高速离心法制备自然杀伤细胞外泌体（NKexo），叶酸（FA）修饰后采用挤压法包载Reo，构建FA-NKexo-Reo递送系统；通过透射电镜（TEM）、纳米粒径分析、蛋白质印迹（WB）法、核磁共振氢谱及流式细胞术等技术表征其理化性质；采用CCK-8、流式细胞术、Transwell实验及激光共聚焦显微镜评估FA-NKexo-Reo递送系统体外细胞毒性及细胞摄取能力；通过人卵巢癌裸鼠皮下移植瘤模型评价FA-NKexo-Reo的肿瘤靶向性、疗效及安全性。结果：FA-NKexo-Reo粒径为（94.0 ± 28.5）nm，Zeta电位为（-21.26 ± 1.57）mV，包封率达（49.7 ± 15.6）%；在中和抗体的存在下，FA-NKexo-Reo对卵巢癌细胞SKOV3和A2780仍可表现出显著的细胞毒性（P < 0.01）；荷瘤鼠活体成像显示FA-NKexo-Reo肿瘤靶向性显著优于NKexo组，肿瘤抑制率提升60%（P < 0.001）。结论：成功制备FA-NKexo-Reo递送系统，在中和抗体的存在下，FA-NKexo-Reo可保护并靶向递送Reo到高表达叶酸受体的卵巢癌细胞，从而增强Reo的抗肿瘤作用。

Objective: To investigate the association between sleep duration and depressive symptoms in children and adolescents, so as to provide scientific evidence for promoting mental health and preventing depression among relevant populations. Methods: A total of 2 192 children and adolescents aged 10-17 years with complete data from the 2018 China Family Panel Studies Database were included. Eight item Center for Epidemiologic Studies Depressive Scale(CES-D8) was used to assess participants depressive levels, and sleep duration was assessed via questionnaire. Multivariate Logistic regression model was used to analyze the association between different sleep duration categories and depressive symptom occurrence among children and adolescents. A restricted cubic spline(RCS) model analyzed the dose response relationship between sleep duration and the risk of depressive symptoms occurrence and segmented Logistic regression models to identify dose response effects. Results: Among the surveyed children and adolescents, 524(23.91%) exhibited depressive symptoms. Compared to those with sufficient sleep, children aged 10-12 years had a higher risk of depressive symptoms on average per day( OR =1.66, 95% CI =1.19-2.33) and during weekdays( OR =1.76, 95% CI =1.26-2.46), as well as in adolescents aged 13-17 years on a daily basis( OR =1.40，95% CI =1.06-1.85) and during weekdays( OR = 1.48，95% CI =1.12-1.95), and excessive sleep in adolescents on rest days was also significantly associated with higher risk of depressive symptoms( OR =1.65，95% CI =1.11-2.43)(all P <0.05). RCS analysis results indicate that children s sleep duration exhibits a linear negative correlation with the risk of depressive symptoms(all P nonlinear >0.05), while adolescents sleep duration showed a U shaped association with depressive symptoms(all P nonlinear <0.05) on a daily basis, during weekdays and weekends, with potential threshold effects at 10.00, 9.88, and 9.60 hours, respectively. Conclusions Sleep duration among children and adolescents is associated with depressive symptoms, with notable age related differeneces. It is recommended to develop targeted age specific interventions to reduce the risk of depressive symptoms in children and adolescents.

Objective To analyze the main causes and risk factors of unplanned reoperation after liver transplantation. Methods The clinical data of 242 liver transplant recipients in the First Affiliated Hospital of Anhui Medical University from January 2015 to December 2024 were retrospectively analyzed. According to whether unplanned reoperation was performed during the same hospitalization after surgery, the recipients were divided into the reoperation group (n=36) and the non-reoperation group (n=206). The preoperative, intraoperative and postoperative data of the two groups, as well as donor and graft-related data, were compared to analyze the risk factors of unplanned reoperation after liver transplantation and the survival status of the two groups. Results Among the 242 liver transplant recipients, 36 underwent unplanned reoperations, with a total of 54 procedures including various laparotomies, endoscopic and interventional surgeries, among which there were 20 laparotomies, 18 endoscopic surgeries and 16 interventional surgeries. The most common cause of unplanned reoperation was biliary complications (20 times), followed by vascular complications (17 times). Compared with the non-reoperation group, the reoperation group had longer graft cold ischemia time, higher postoperative fatality rate of recipients, longer length of stay in the intensive care unit and postoperative hospital stay, and higher total hospitalization costs (all P<0.05). The incidence of unplanned reoperation was higher in recipients who underwent split liver transplantation (P<0.05). Multivariate analysis showed that intraoperative blood loss ≥1 000 mL, positive culture of graft perfusate and split liver transplantation were independent risk factors for unplanned reoperation (all P<0.05). The postoperative 7-day, 1-month, 3-month and 6-month survival rates of recipients in the reoperation group and the non-reoperation group were 100% vs. 98.1%, 88.9% vs. 94.2%, 69.4% vs. 90.8% and 66.7% vs. 90.8%, respectively, and the postoperative survival rate of recipients in the reoperation group was lower than that in the non-reoperation group (P<0.05). Conclusions The main causes of unplanned reoperation after liver transplantation are biliary complications, vascular complications, abdominal incision infection and intra-abdominal hemorrhage. Intraoperative massive blood loss, positive culture of graft perfusate and split liver transplantation are the risk factors associated with unplanned reoperation after liver transplantation.

Objective To investigate the epidemiological characteristics of lymphoma mortality and potential years of life lost among Wuhan residents, and to provide a scientific basis for the prevention and control of lymphoma. Methods Data on lymphoma deaths among residents in Wuhan from 2010 to 2019 were collected from the population-based Wuhan Mortality Surveillance System. Joinpoint regression model was used to evaluate the trends in age-standardized mortality rates and potential years of life lost due to lymphoma among the residents of different ages and genders. Potential years of life lost rate was used to assess the disease burden. Results There were 4 055 deaths (2481in male and 1 574 in female) from major kidney diseases in Wuhan residents between 2010 to 2019, with an age-standardized mortality rate of 5.11/100 000. The mortality rate of major kidney diseases showed an upward trend with increasing age. Between 2010 and 2019, the age-standardized mortality rates of the multiple myeloma and non-Hodgkin lymphoma were significantly increased (APC=6.924%, APC=1.407%, P＜0.05), and the potential years of life lost rate of non-Hodgkin lymphoma in female were significantly increased (APC=7.935%，P＜0.05). Conclusion From 2010 to 2019, the mortality rate of lymphoma among residents of Wuhan City shows an upward trend, especially for multiple myeloma and non-Hodgkin lymphoma. The disease burden of multiple myeloma shows an increasing trend in the female population, suggesting the need to take corresponding preventive and control measures.

Objective To investigate whether long non-coding RNA(lncRNA)myocardial infarct-associat-ed transcription factor(MIAT)promotes sepsis-induced acute kidney injury(AKI)by regulating microRNA-338-3p(miR-338-3p)/platelet thromboplastin-1(THBS1)axis.Methods Septic AKI model was established by cecal ligation and puncture.The rats were divided into control group and sepsis AKI group,with 10 rats in each group.The expression levels of lncRNA MIAT,miR-338-3p and THBS1gene in renal tissue were detec-ted by quantitative real-time PCR(qPCR).The levels of serum urea nitrogen(BUN)and creatinine(Cre)were detected by automatic biochemical analyzer.Renal tubular epithelial NRK-52E cells were induced by li-popolysaccharide(LPS)to establish a cell model of sepsis induced AKI.NRK-52E cells were divided into CK group,LPS group,LPS+si-NC group,LPS+si-lncRNA MIAT group,LPS+si-lncRNA MIAT+inhibitor NC group,LPS+si-lncRNA MIAT+miR-338-3p inhibitor group,LPS+si-lncRNA MIAT+oe-NC group,LPS+si-lncRNA MIAT+oe-THBS1 group.qPCR was used to detect the expression levels of lncRNA MIAT,miR-338-3p and THBS1 gene in each group.Cell counting kit 8 was used to detect cell proliferation in each group.Flow cytometry was used to detect the apoptosis rate of each group.Enzyme-linked immunosorbent assay was used to detect the levels of lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-10,malondialdehyde(MDA)and the activities of superoxide dismutase(SOD)and catalase(CAT)in each group.The expression of NOD-like receptor pyrin domain-containing protein 3(NLRP3),Caspase-1,cleaved caspase-3 and THBS1 protein were detected by Western blot.The targeting relationship between miR-338-3p and lncRNA MIAT and THBS1 was verified.Results The expression levels of BUN,Cre,lncRNA MI-AT and THBS1 gene were increased(P<0.05),and the expression level of miR-338-3p was decreased in sep-sis AKI group(P<0.05).Compared with CK group,the expression of lncRNA MIAT,THBS1 gene,apopto-sis rate and the levels of IL-6,LDH,TNF-αand MDA were significantly increased(P<0.05),the protein ex-pression levels of NLRP3,Caspase-1,cleaved caspase-3 and THBS1 were significantly increased(P<0.05),while the expression of miR-338-3p,A450(24,48 h)value and IL-10 level were significantly decreased(P<0.05),and the activities of CAT and SOD were significantly decreased in LPS group and LPS+si-NC group(P<0.05).Compared with LPS+si-NC group,the expression of lncRNA MIAT,THBS1 gene,apoptosis rate and the levels of IL-6,LDH,TNF-α and MDA were significantly decreased(P<0.05),the protein expression levels of NLRP3,Caspase-1,cleaved caspase-3 and THBS1 were significantly decreased(P<0.05),while the expression of miR-338-3p,A450(24,48 h)value and IL-10 level were significantly increased(P<0.05),and the CAT and SOD activities were significantly increased in LPS+si-lncRNA MIAT group(P<0.05).Silen-cing miR-338-3p expression or up-regulation of THBS1 gene expression could attenuate the improvement effect of lncRNA MIAT on sepsis AKI(P<0.05).Conclusion lncRNA MIAT promotes sepsis-induced AKI through regulating miR-338-3p/THBS1 axis.

Objective:To investigate the correlation between lymphocyte-monocyte ratio (LMR) and long-term prognosis after distal cholangiocarcinoma.Methods:A retrospective case-control study was conducted to analyze the clinical data of 186 patients with distal cholangiocarcinoma who underwent radical pancreaticoduodenectomy at Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2013 to December 2023. Among them, there were 109 males and 77 females, with an age of (65.4±9.4) years, ranging from 29 to 85 years. The data of preoperative peripheral blood routine examination of the patients were collected, and the patients were divided into a high LMR group(LMR>2.98, n=100) and a low LMR group(LMR≤2.98, n=86). The preoperative, intraoperative and postoperative clinical characteristics of the two groups were compared. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), and t-test was used for inter-group comparison. Measurement data with non-normal distribution were expressed as M( Q1, Q3), and Mann-Whitney U test was used for inter-group comparison. Chi-square test was used for inter-group comparison of count data. The Cox proportional hazards regression model was used for univariate and multivariate prognostic analysis, and the Kaplan-Meier estimation method was used to create survival curves to analyze and evaluate the influencing factors of long-term prognosis after distal cholangiocarcinoma surgery. Results:Univariate analysis results showed that gender, age, BMI, history of diabetes, carcinoembryonic antigen; operation duration, intraoperative blood loss; resection margin status, degree of tumor cell differentiation, and presence of postoperative complications had no statistically significant differences in their impact on the prognosis of patients after distal cholangiocarcinoma surgery( P>0.05). In contrast, LMR, neutrophilto-lymphocyte ratio, platelet-lymphocyte ratio, albumin, total bilirubin, carbohydrate antigen 199, intraoperative blood transfusion, tumor diameter, and lymph node metastasis showed statistically significant differences in their influence on the postoperative prognosis of distal cholangiocarcinoma patients( P<0.05). Multivariate analysis results indicated that LMR≤2.98( HR=1.776, 95% CI: 1.153-2.736), CA19-9>37 U/mL( HR=1.521, 95% CI: 1.025-2.259), and lymph node metastasis( HR=1.601, 95% CI: 1.106-2.318) were independent risk factors affecting patient prognosis( P<0.05). The 1-, 3-, and 5-year cumulative survival rates in the high LMR group were 91%, 40%, and 20% respectively, while those in the low LMR group were 58.1%, 15.1%, and 8.1% respectively, with a statistically significant difference( P<0.05). Conclusion:Preoperative LMR for distal cholangiocarcinoma can suggest a long-term prognosis, and a low LMR value suggests a poor prognosis.

Objective To analyze the risk factors of postpartum mild uterine prolapse and to investigate the effects of Buzhong Yiqi Decoction on serum transforming growth factor β1(TGF-β1),matrix metalloproteinase 2(MMP-2),and microRNA-30d(miRNA-30d).Methods By blinded and controlled methods and through convenience sampling,80 postpartum patients with mild uterine prolapse were retrospectively collected from Tangshan Maternal and Children Health Hospital from January 2020 to January 2022,and they served as the uterine prolapse group.In addition,80 postpartum patients without uterine prolapse from the same department of the hospital in the same period served as the healthy group.Questionnaire investigation was performed for collecting the basic information such as delivery manner,age,parity,gestational weeks,neonatal weight and body mass index(BMI).The risk factors of postpartum uterine prolapse were investigated by univariate analysis and multivariate logistic regression analysis.The patients in the uterine prolapse group were then divided into the observation group and the control group according to the medication of Buzhong Yiqi Decoction or not,with 40 cases in each group.The control group was treated with electromyographic biofeedback therapy(EBT),while the observation group was treated with Buzhong Yiqi Decoction combined with EBT,and the course of treatment for the two groups covered one month.The changes of pelvic floor function and levels of serum TGF-β1,MMP-2,miRNA-30d in the patients of the two groups were observed before and after treatment.After treatment,the clinical efficacy of patients in the two groups was evaluated.Results(1)The results of univariate analysis showed that compared with the healthy group,the patients in the uterine prolapse group were elder,usually had given two childbirths or more by vaginal delivery,and accounted for a high percentage of overweight/obesity(BMI≥24.0 kg/m2)and perineal laceration,the difference being statistically significant(P<0.05 or P<0.01).(2)The results of multivariate logistic regression analysis showed that age,delivery manner,parity,laceration of perineum,and BMI were risk factors for postpartum uterine prolapse(P<0.01).(3)After one month of treatment,the total effective rate of Buzhong Yiqi Decoction in the observation group was 95.00%(38/40),while that in the control group was 77.50%(31/40),and the intergroup comparison(tested by chi-square test)showed that the efficacy of the observation group was significantly superior to that of the control group(P<0.05).(4)The observation of effects of Buzhong Yiqi Decoction on pelvic floor function showed that after treatment,the distance between the cervix and the hymenal edge,the maximum intravaginal pressure,the voltage of vaginal muscles,the contraction time of pelvic floor muscles of typeⅠand Ⅱmuscle fibers in the two groups of patients were significantly improved compared with those before treatment(P<0.05),and the improvement in the observation group was significantly superior to that in the control group(P<0.01).(5)The observation of effects of Buzhong Yiqi Decoction on serological indicators showed the post-treatment serum TGF-β1 levels in the two groups were increased compared with those before treatment(P<0.05),and the serum levels of MMP-2 and miRNA-30d were decreased compared with those before treatment(P<0.05),and the increase of serum TGF-β1 level and the decrease of serum MMP-2 and miRNA-30d levels in the observation group were both significantly superior to those in the control group(P<0.01).Conclusion The incidence of postpartum uterine prolapse is related to age,delivery manner,parity,laceration of perineum,BMI,etc.The treatment of Buzhong Yiqi Decoction on the basis of EBT can improve the pelvic floor function of the patients and regulate the expression of serum TGF-β1,MMP-2,and miRNA-30d,and the clinical efficacy is remarkable.

ObjectiveTo evaluate the efficacy and safety of transcutaneous electrical acupoint stimulation (TEAS) for muscle atrophy in patients with immobilization after surgical fixation of foot and ankle fractures.MethodsThis was a two-arm randomized controlled trial wherein 80 patients were recruited and divided into control (n = 40) and intervention (n = 40) groups. The control group received conventional orthopedic treatment, whereas the intervention group received TEAS and conventional treatment. The intervention group received TEAS 3 times a week for 30 min each time for 8 weeks. The primary outcomes were muscle thickness (MT) and cross-sectional area (CSA) of the rectus femoris and gastrocnemius muscles, whereas the secondary outcome measure was echo intensity (EI). Data were collected before the fixation operations (baseline assessment) and 4 and 8 weeks after intervention.ResultsCompared with baseline, the MT and CSA were reduced in both groups by the end of treatment, whereas EI increased in both groups. At week 4, the reduction in the rectus femoris CSA in the intervention group was significantly lower than that in the control group (P = .02); however, the between-group differences in the MT and EI (all P .05) were not significant. No serious adverse events were observed in either group.ConclusionOur study showed that TEAS can improve muscle atrophy by attenuating the decline in the muscle CSA. Because this was only a pilot trial, subsequent studies will need longer follow-ups and larger sample sizes.

BACKGROUND:Psoralen has a strong anti-osteoporotic activity and may have a restorative effect on chemotherapy-induced osteoporosis. OBJECTIVE:To explore the restorative effect of psoralen on the osteogenic differentiation of bone marrow mesenchymal stem cells in mice inhibited by cyclophosphamide and its mechanism. METHODS:C57BL/6 mouse bone marrow mesenchymal stem cells were isolated and cultured.Effect of psoralen on viability of bone marrow mesenchymal stem cells was detected by MTT assay.Osteogenic induction combined with alkaline phosphatase staining was used to determine the optimal dose of psoralen to restore the osteogenic differentiation of bone marrow mesenchymal stem cells inhibited by cyclophosphamide.The mRNA expression levels of Runx2,alkaline phosphatase,Osteocalcin,osteoprotegerin,and Wnt/β-catenin signaling pathway-related genes Wnt1,Wnt4,Wnt10b,β-catenin,and c-MYC were measured by RT-qPCR at different time points under the intervention with psoralen.The protein expression of osteogenic specific transcription factor Runx2 and Wnt/β-catenin signaling pathway related genes Active β-catenin,DKK1,c-MYC,and Cyclin D1 was determined by western blot assay at different time points under the intervention with psoralen. RESULTS AND CONCLUSION:(1)There was no significant effect of different concentrations of psoralen on the viability of bone marrow mesenchymal stem cells.The best recovery of the inhibition of osteogenic differentiation of bone marrow mesenchymal stem cells caused by cyclophosphamide was under the intervention of psoralen at a concentration of 200 μmol/L.(2)Psoralen reversed the reduction in osteogenic differentiation marker genes Runx2,alkaline phosphatase,Osteocalcin and osteoprotegerin mRNA expression and Runx2 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(3)Psoralen reversed the decrease in Wnt/β-catenin pathway-related genes Wnt4,β-catenin,c-MYC mRNA and Active β-catenin,c-MYC,and Cyclin D1 protein expression and the increase in DKK1 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(4)The results showed that cyclophosphamide inhibited osteogenic differentiation of bone marrow mesenchymal stem cells in mice,and psoralen had a restorative effect on it.The best intervention effect was achieved at a concentration of 200 μmol/L psoralen,and this protective effect might be related to the activation of Wnt4/β-catenin signaling pathway by psoralen.

BACKGROUND:Inflammation affects the osteogenic differentiation of periodontal ligament stem cells,and the osteogenic ability and autophagy level of periodontal ligament stem cells are closely related.However,there are no relevant reports on whether inflammation affects the osteogenic ability and autophagy level of periodontal ligament stem cells at different stages of osteogenic differentiation. OBJECTIVE:To explore alkaline phosphatase expression and autophagy periodontal ligament stem cells levels in periodontitis and normal conditions. METHODS:Periodontal ligament stem cells from normal and periodontitis patients were isolated and cultured,and underwent Vimentin,pan-CK,and Stro-1 fluorescence staining.At 3,7,and 14 days of osteogenic differentiation,western blot assay was used to detect the protein expression levels of alkaline phosphatase,LC3B,Beclin1,and ATG5 in normal and inflammatory periodontal ligament stem cells.The mRNA expression levels of alkaline phosphatase,bone sialoprotein,osteocalcin,Runx2,LC3B,Beclin1,and ATG5 were detected by real-time PCR. RESULTS AND CONCLUSION:(1)Stro-1 was positive,Vimentin was positive,and pan CK was negative in periodontal ligament stem cells.(2)At 3,7,and 14 days after osteogenic differentiation,compared with normal periodontal ligament stem cells,the mineralization nodules formed by periodontal ligament stem cells from inflammatory sources were significantly reduced(P<0.01);the expression of alkaline phosphatase protein and mRNA was significantly lower(P<0.05);the mRNA expression levels of bone sialoprotein,osteocalcin,and Runx2 were significantly decreased(P<0.05).(3)At 7 and 14 days after osteogenic differentiation,compared with normal periodontal ligament stem cells,the expression levels of ATG5,LC3B,and Beclin1 proteins and mRNA of periodontal ligament stem cells were downregulated(P<0.05).These findings suggest that inflammation reduces the activity of periodontal ligament stem cells in mineralizing nodule formation and the expression of alkaline phosphatase and weakens the autophagy potential of periodontal ligament stem cells at 7 and 14 days after osteogenic differentiation.