BACKGROUND:Chondrocyte apoptosis is an important factor in the development of osteoarthritis,and Complanatoside A has a flavonoid effect,which can inhibit apoptosis of various cells,but its effect on chondrocyte apoptosis and the mechanism of action are not clear. OBJECTIVE:To investigate the intrinsic association and mechanism of Complanatoside A in chondrocyte apoptosis based on the Wnt/β-catenin signaling pathway. METHODS:(1)The cartilage tissues of the femur and tibia transected during knee arthroplasty were collected,and chondrocytes were isolated,cultured in vitro,and identified.(2)Cell counting kit-8 was used to detect the optimal intervention concentration of Complanatoside A in the concentration range of 0-160 μmol/L.(3)Chondrocytes were divided into blank group,sodium nitroprusside(1.5 mmol/L)-induced group,and sodium nitroprusside(1.5 mmol/L)+Complanatoside A(5 μmol/L)group.The viability and apoptosis rate of the cells in each group were detected by cell counting kit-8 and flow cytometry.The expression of type Ⅱ collagen and SOX9 was detected by immunofluorescence staining.The expression of apoptosis-related proteins and Wnt/β-catenin pathway proteins was detected by western blot assay. RESULTS AND CONCLUSION:The cells extracted in vitro were cultured and stained,and were clearly identified as chondrocytes.Complanatoside A had no obvious cytotoxicity to chondrocytes in the concentration range of 0-80 μmol/L,and significantly improved the chondrocyte viability in the concentration range of 2.5-10 μmol/L,especially when the concentration was 5 μmol/L.The apoptotic rate of chondrocytes was higher in the sodium nitroprusside-induced group than the blank control group,while the apoptotic rate was lower in the sodium nitroprusside+Complanatoside A group than the sodium nitroprusside-induced group.The fluorescence intensity of type Ⅱ collagen and SOX9 in chondrocytes was weaker in the sodium nitroprusside-induced group than the blank control group,while the fluorescence intensity of type Ⅱ collagen and SOX9 in the sodium nitroprusside+Complanatoside A group was higher than that of the sodium nitroprusside-induced group.In the sodium nitroprusside-induced group,the protein expression of Bax,Caspase-3,matrix metalloproteinase 13,Wnt3a,Wnt5a and β-catenin was higher than that of the blank control group,while the protein expression of Bcl-2 was lower than that of the blank control group.In the sodium nitroprusside+Complanatoside A group,except for the protein expression of Bcl-2 which was higher than that of the sodium nitroprusside-induced group,the expression of the other aforementioned proteins was lower than that of the sodium nitroprusside-induced group.To conclude,Complanatoside A has a certain inhibitory effect on chondrocyte apoptosis,which could regulate apoptosis-related proteins and promote the expression of chondrocyte regulatory factors,and presumably might play a role through inhibiting the Wnt/β-catenin signaling pathway.

Objective:To investigate the efficacy of Shuotong ureteroscope combined with flexible ureteroscope in the treatment of 2-3 cm lower calyceal calculi, and analyze the influencing factors.Methods:A total of 102 patients with lower calyceal calculi were treated in the Second People′s Hospital of Yulin from February 2019 to December 2022, and they were divided into the observation group and the control group, with 51 cases in each group. The patients of the observation group were treated with Shuotong ureteroscope combined with flexible ureteroscope, while the patients of the control group were treated with flexible ureteroscope. According to whether the stones were completely removed after operation, all patients were divided into non-stone removal group ( n=13) and stone removal group ( n=89). The operation time, hospitalization time, lithotripsy time, intraoperative blood loss, complication rate and stone clearance rate were compared between the observation group and the control group. Generalized Estimation Equation was used to analyze and evaluate the effects of treatment time, treatment scheme and their interaction on visual analogue scale (VAS), white blood cell (WBC), blood urea nitrogen (BUN), blood creatinine (Cr), hemoglobin (HGB) and procalcitonin (PCT). Univariate and multivariate Logistic regression were used to analyze the risk factors of stone removal rate. Nomogram model was constructed based on risk factors and evaluate the model. Results:Compared with the control group, operation time [(118.72±9.61) min vs (136.65±11.27) min], hospitalization stay [(6.43±1.12) d vs (10.29±2.23) d] and the lithotripsy time [ (51.23±10.38) min vs (56.62±11.43) min] of the observation group were shorter, and the amount of intraoperative blood loss [(128.52±10.20) mL vs (157.53±15.31) mL] were significantly less than those of the control group ( P< 0.05). The results of Generalized Estimation Equation analysis showed that treatment time, treatment regimen and their interaction had significant effects on WBC, HGB, BUN, Cr, PCT and VAS ( P< 0.05). Compared with the control group, the incidence of complications (5.88% vs 19.61%) of the observation group was lower and the stone clearance rate ( 94.12% vs 80.39%) was significantly higher than those in the control group ( P< 0.05). The mode of operation, infundibulopelvic angle(IPA), caliceal pelvic height (CPH) and the maximum diameter of stones were all influencing factors of stone removal rate in patients with 2-3 cm lower calyceal calculi. The nomogram model constructed in this study has good differentiation, calibration and clinical practicability, and can better identify high-risk patients with incomplete removal of 2-3 cm lower calyceal calculi. Conclusions:Shuotong ureteroscope combined with flexible ureteroscope is a safe, effective method for the treatment of 2-3 cm lower calyceal calculi. It has the advantages of simple operation, less intraoperative bleeding, less postoperative complications and high stone clearance rate. IPA, CPH, the maximum diameter of calculi and the mode of operation were all independent factors affecting the stone clearance rate of 2-3 cm lower calyceal calculi. The nomogram model constructed in this study can well identify the high-risk patients with incomplete clearance of 2-3 cm lower calyceal calculi.

Objective:To investigate the value of mind map combined with blended teaching in improving the teaching quality of medicinal botany. Methods:A total of 102 students studying the course of medicinal botany were enrolled as subjects. The 51 students in the class of 2020 were enrolled as control group and received conventional teaching, and the 51 students in the class of 2021 were enrolled as observation group and received mind map combined with blended teaching. The two groups were assessed in terms of examination scores, critical thinking ability scores, self-learning ability scores, and student feedback on teaching quality. SPSS 22.0 was used for the t-test and the chi-square test. Results:Compared with the control group, the observation group had significantly higher examination scores ( t=3.01 and 3.14, P=0.003 and 0.002). After practice, both groups had increases in the scores of critical thinking ability and self-learning ability, and the observation group had significantly higher scores than the control group ( t=11.22 and 2.69, P<0.001 and P=0.008). Compared with the control group, the observation group had a better student feedback on teaching quality than the control group ( t=6.79, 7.83, 7.26, 7.43, and 8.54, P=0.009, 0.005, 0.007, 0.006, and 0.003). Conclusion:The combination of mind map and blended teaching can improve the examination scores of students and their critical thinking ability and self-learning ability, and students believe that this teaching model can help to improve teaching quality.

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3-LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.

OBJECTIVE:To investigate the effects of Tongxinluo capsules on the levels of serum cyclophilin A (CyPA) and matrix metalloproteinase-9 (MMP-9) in patients with coronary heart disease (CHD) after percutaneous coronary intervention (PCI).METHODS:A total of 115 CHD patients undergoing PCI were randomly divided into control group (59 cases) and observation group (56 cases).Control group was given conventional treatment.Observation group was additionally given Tongxinluo capsules 0.78 g,3 times a day,for consecutive 6 months,on the basis of control group.The levels of CyPA and MMP-9 as well as the occurrence of ADR were observed in 2 groups before and after treatment.RESULTS:Before treatment,there was no statistical significance in serum level of CyPA and MMP-9 between 2 groups (P＞0.05).The levels of CyPA in 2 groups 1 d and 1 month after surgery as well as the level of CyPA in control group 6 months after surgery were all significantly higher than before surgery,but the observation group was significantly lower than the control group 1 and 6 months,this index was decreased gradually as time,with statistical significance (P＜0.05).There was no statistical significance in the levels of CyPA between 2 groups 1 d after surgery or in observation group between 6 months after surgery and before surgery (P＞0.05).The levels of MMP-9 in 2 groups 1 d and 1 month after surgery were significantly higher than before surgery,and the observation group was significantly lower than the control group,this index was decreased gradually as time,with statistical significance (P＜0.05).There was no statistical significance in the levels of MMP-9 of 2 groups between 6 months after surgery and before surgery(P＞0.05).There was no statistical significance in the incidence of ADR between 2 groups (P＞0.05).CONCLUSIONS:Based on routine treatment,Tongxinluo capsules can significantly reduce the levels of CyPA and MMP-9 in CHD patients after PCI.

BACKGROUND:Liver regeneration is the key factor influencing the prognosis of living donor liver transplantation. There has not been the research on special miRNA of liver regeneration after living donor liver transplantation. OBJECTIVE:To analyze the variation of miRNAs expression profile after rat reduced-size liver transplantation at certain time point, select and verify target miRNA which can provide targeting intervention strategies in liver regeneration after rat reduced-size liver transplantation and provide theoretical evidence for liver regeneration after living donor liver transplantation.METHODS:The reduced-size liver transplantation models were established. miRNAs microarray was used to detect miRNA expression. In differentialy expressed microRNAs, real-time quantitative PCR was utilized to detect target miRNAs. The credibility of miRNAs microarray results was verified. RESULTS AND CONCLUSION:Compared with rat liver tissue in the sham operation group, 11 miRNAs up-regulated in reduced-size liver transplantation, including let-7b-5p, let-7c-5p, miR-101a-3p, miR-103-3p, miR-130a-3p, miR-142-5p, miR-186-5p, miR-199a-3p, miR-21-5p, 221-3p and miR-34a-5p. Four miRNAs were down-regulated, including miR-26b-5p, miR-150-5p, miR-19a-3p and rno-miR-146-5p. PCR test further verified that miR-221-3p and miR-199a-3p expression changes approximated the chip results at 24, 48 hours and 1 week, indicating that results of miRNA microarray were believable. These results verified that it exists variation of miRNAs expression profile after rat reduced-size liver transplantation, which picked out and verified the target miRNAs.

Objective To evaluate the association between birth weight and subsequent risk of type 2 diabetes (T2DM) across two generations in Chinese females.Methods A cross-sectional survey was conducted among 10 324 blood-related females in Shanghai from November 2012 to January 2013.A structured questionnaire was used to collect data about demographic information,birth weight,lifestyle factors and diagnosis of T2DM.Path analysis was performed to estimate the matemal effect of birth weight and diabetes status on their offspring,and mediation analysis was conducted to test whether birth weight mediates the association between maternal and daughters' diabetes status.Results A positive association was observed between maternal and daughters' birth weight.One kg increase in maternal birth weight was associated with an average of 0.335 (95％CI:0.307-0.363) kg elevated birth weight in female offspring.Maternal diabetes status was also related with an average of 0.066 (95％ CI:0.025-0.107) kg increase in birth weight and tripled risk of subsequent T2DM (OR=3.173,95％CI:1.946-5.174) in female offspring.In all subjects aged ≥20 years,no significant relationship was observed between birth weight and risk of T2DM subsequently after adjusting for age.Offspring' s birth weight,as a mediator of maternal and daughters' diabetes status,explained 2.8％ of their association.Conclusion Maternal birth weight and diabetes status may influence their female offspring' s birth weight and subsequent risk of T2DM.

Objective To study the efficacy of medication management, symptom management and community reentry skills-training modules to prevent rural patients with schizophrenia from relapse and increase their social function.Methods 89 subjects were randomly assigned to the skills training group (45 cases) and the control group (44 cases). Both groups received the same treatments, but the skills training courses were given to the skills training group for twelve weeks when the subjects were addmitted into the trial and at their one-year follow-up timepoint respectively. And a two-year follow-up was carried out. All subjects were evaluated with PANSS, SDSS, relapse and employeement.Results 89 subjects had completed the study. The skills training group demonstrated clinical Results significantly superior to those of the control group on overall improvement according to PANSS and SDSS. Substantially, statistically significant advantages of the skills training group were related to 1) the rate of relapse (11.1% vs 31.8%), 2) the rate of employment (37.8% vs 15.9%).Conclusion Medication management, symptom management and community-re-entry skills-training modules are effective in reducing patients' psychiatric symptoms, preventing them from relapse and increasing their social function.

Objective: To select suitable conditions for prokaryotic expression and purification of rhIL-17. Methods: rhIL-17 was expressed in E. coli host under heat induction. After compared among the expression amounts in different media under different heat induction time, the most suitable conditions was selected. The target protein was present in the form of inclusion body. The precipitate of inclusion was obtained and purified after 6M guanidine solublization or 2% SDS solublization. Results: Either protocol could yield rhIL-17 with high purity and stable activity. The SDS solublization mehthod gives rise to much more higher productivity than the guanidine solublization method. Conclusion: rhIL-17 were expression in E. coli system and purified to homogenicity by SDS solublization methods with high productivity.

To investigate effects of rhIL-17 on growth and development of mouse bone marrow progenitors andhuman cord blood LD34~+ stem cells. Methods: Mouse bone marrow progenitors were isolated by routine protocol, and CD34~+ stem cells were isolated from normal human cord blood by Mini-MACS, then cultured with rhIL-17 and/or GM-CSF/IL-4. The phenotypes of the cells were analyzed by FACS, IL-12 level was analyzed by ELISA and T cell stimulating activity in allo-MLR was determined by [~3H]-TdR incorporation. Results: Expression of MHC class II molecules and B7-2 on the surface of immature DC derived from mouse bone marrow progenitors was up-regulated by IL-17. The capacity of the cells to secrete IL-12 and their T cell stimulating activity were also enhanced. The cells showed the characteristics of mature DC. After cultured with IL-17 for 9 days, the number of CD34~+ stem cells increased by 2 times. The phenotypes of some cells were CDla~(high), B7-2~(high), and HLA-DR~(lwo). The cells could stimulate allo geneic T cells to proliferate but their capacity was lower than that of the cells cultured with IL-17 combined with GM-CSF. The cells cultured with IL-17 and GM-CSF proliferated markedly and the rate of CDla~+ and B7-2~+ cells increased significantly. The T cell stimulating activity of cells was also augmented. Conclusion: IL-17 could promote DC derived from mouse bone marrow progenitors to mature. When combined with GM-CSF, IL-17 could induce human CD34~+ stem cells not only to proliferate markedly but also to show characteristics of DC, indicating that CD34~+ stem cells might differentiate to DC by IL-17.