Objective:To explore the relationship between insomnia and osteoporosis.Methods:Mendelian randomization (MR) analysis were used in this study. The single nucleotide polymorphisms (SNPs) related to insomnia from genome-wide association analysis research data were selected as the instrumental variables by using inverse variance weighted (IVW), MR-Egger regression, weighted median method, maximum likelihood, penalized weighted median estimator, and Mendelian randomization robust adjusted profile score (MR-RAPS) to determine the causal relationship between insomnia and osteoporosis. Odds ratio ( OR) and 95% confidence interval ( CI) values were used to evaluate the association between insomnia and osteoporosis. Cochran′s Q-test was used to detect heterogeneity of SNPs, MR-Egger regression was used to test for level pleiotropy, and the leave-one-out method was used to test sensitivity, MR pleiotropy residual sum and outlier (MR-PRESSO) method and radial MR were used to detect erroneous outliers. Results:The screening criteria were set based on the three major assumptions of MR; finally, 31 SNPs were included in the MR analysis. The results of MR causal effect analysis using the IVW method showed that insomnia increased the risk of osteoporosis by about 0.7% ( OR=1.007, 95% CI 1.001-1.014, P=0.044); heterogeneity testing showed heterogeneity between SNPs ( Q=57.91, P<0.001); and the MR- Egger intercept test did not indicate horizontal pleiotropy in this study (intercept value=3.807×10 -5, P=0.888). Leave-one-out method showed that no single SNP had a significant impact on the overall results. No abnormal SNP was detected according to the MR-PRESSO results ( P=0.059), and radial MR did not detect any outliers. Conclusion:Mendelian randomization analysis showed that insomnia can increase the risk of osteoporosis.

Objective To investigate the effect of Sclareol(SCL)on LX-2 hepatic stellate cell activation and CCl4-induced liver fibrosis in mice,and to further explore its mechanism.Methods A total of 40 Kunming mice were randomly divided into healthy group,model group(10%CCl4)and SCL administration group,and silybin positive control group(10%CCl4+100 mg·kg-1 Silybin),and SCL administration group was divided into low SCL(10%CCl4+20 mg·kg-1 SCL)and high dose group(10%CCl4+40 mg·kg-1 SCL).Mice in all groups were intraperitoneally injected with 10%olive oil-diluted CCl4 three times a week for four weeks,except for the healthy group.Starting from the third week,the dosing group was given different doses of SCL by gavage daily,and the positive control group was given silybin daily,and the mice were sacrificed and serum and liver tissue were collected after four weeks.In whole animal experiments,biochemical kits were used to detect the changes in the serum levels of glutamate aminotransferase(ALT)and aspartate aminotransferase(AST)in mice with liver fibrosis.Hematoxylin-eosin(HE),Sirius Red and Masson staining were used to detect microstructural changes and collagen deposition in liver tissues.Immunohistochemistry was used to detect the expression of fibrosis marker proteins α-smooth muscle actin(α-SMA)and fibrous collagen I.in liver tissues.In vitro,LX-2 human hepatic stellate cells were used for normal culture in the blank group,and the activation of LX-2 hepatic stellate cells was induced by transforming growth factor-β1(TGF-β1)in the model group,and the SCL administration group was divided into SCL low-dose group(5 ng·mL-1 TGF-β1+10 μmol·L-1 SCL)and high-dose group(5 ng·mL-1 TGF-β1+20 μmol·L-1 SCL).Subsequently,Transwell and EdU assays were used to detect the effects of SCL on the migration and proliferation of LX-2 cells.The expression of fibrosis marker proteins α-SMA and Collagen I.affected by SCL was detected by immunofluorescence.Western blot was used to detect the expression of related proteins in TGF-β/Smad pathway.Results In animal experiments,compared with the model group,SCL could significantly improve the liver function indexes and liver histopathological changes in liver fibrosis model mice.In addition,in vitro cell experiments,compared with the model group,SCL can effectively inhibit the migration and proliferation of hepatic stellate cells and inhibit their activation.Further studies showed that compared with the model group,SCL significantly up-regulated the expression of Smad7 protein and significantly down-regulated the phosphorylation levels of Smad2 and Smad3 proteins.Conclusion SCL has a significant alleviating effect on CCl4-induced liver fibrosis and TGF-β1-induced LX-2 activation in mice,and the mechanism may be related to the regulation of TGF-β/Smad pathway.

Objective To investigate the effect of Sclareol(SCL)on LX-2 hepatic stellate cell activation and CCl4-induced liver fibrosis in mice,and to further explore its mechanism.Methods A total of 40 Kunming mice were randomly divided into healthy group,model group(10%CCl4)and SCL administration group,and silybin positive control group(10%CCl4+100 mg·kg-1 Silybin),and SCL administration group was divided into low SCL(10%CCl4+20 mg·kg-1 SCL)and high dose group(10%CCl4+40 mg·kg-1 SCL).Mice in all groups were intraperitoneally injected with 10%olive oil-diluted CCl4 three times a week for four weeks,except for the healthy group.Starting from the third week,the dosing group was given different doses of SCL by gavage daily,and the positive control group was given silybin daily,and the mice were sacrificed and serum and liver tissue were collected after four weeks.In whole animal experiments,biochemical kits were used to detect the changes in the serum levels of glutamate aminotransferase(ALT)and aspartate aminotransferase(AST)in mice with liver fibrosis.Hematoxylin-eosin(HE),Sirius Red and Masson staining were used to detect microstructural changes and collagen deposition in liver tissues.Immunohistochemistry was used to detect the expression of fibrosis marker proteins α-smooth muscle actin(α-SMA)and fibrous collagen I.in liver tissues.In vitro,LX-2 human hepatic stellate cells were used for normal culture in the blank group,and the activation of LX-2 hepatic stellate cells was induced by transforming growth factor-β1(TGF-β1)in the model group,and the SCL administration group was divided into SCL low-dose group(5 ng·mL-1 TGF-β1+10 μmol·L-1 SCL)and high-dose group(5 ng·mL-1 TGF-β1+20 μmol·L-1 SCL).Subsequently,Transwell and EdU assays were used to detect the effects of SCL on the migration and proliferation of LX-2 cells.The expression of fibrosis marker proteins α-SMA and Collagen I.affected by SCL was detected by immunofluorescence.Western blot was used to detect the expression of related proteins in TGF-β/Smad pathway.Results In animal experiments,compared with the model group,SCL could significantly improve the liver function indexes and liver histopathological changes in liver fibrosis model mice.In addition,in vitro cell experiments,compared with the model group,SCL can effectively inhibit the migration and proliferation of hepatic stellate cells and inhibit their activation.Further studies showed that compared with the model group,SCL significantly up-regulated the expression of Smad7 protein and significantly down-regulated the phosphorylation levels of Smad2 and Smad3 proteins.Conclusion SCL has a significant alleviating effect on CCl4-induced liver fibrosis and TGF-β1-induced LX-2 activation in mice,and the mechanism may be related to the regulation of TGF-β/Smad pathway.

Objective:Observed activity of adjuvant arthritis ( AA) rats Hedgehog signaling pathway and explore the effect of Cyclopamine on rat arthritis and kidney injury.Methods:40 rats were randomly divided into control group,Cyclopamine group,AA+Cyclopamine group and AA model group.We used Freund′s complete adjuvant rat model with Cyclopamine intraperitoneal injection.By measuring paw swelling, systemic inflammation and arthritis semi-quantitative assessment methods of rats to evaluate the model.HE staining was used to detect the renal pathological changes of rats.Western blot was used to detect kidney Gli1 protein expression levels of the rats.Immunohistochemical staining was used to detect the rat kidney TNF-α,IFN-γ,IL-6 expression.Results:After using Cyclo-pamine,the AA rat paw swelling reduced and arthritis refers relieved significantly.Compared with the control group,Cr,BUN and organ coefficient increased significantly in AA model group, and the difference was statistically significant.Meanwhile renal TEM detection appeared obvious pathological changes in rats of AA model group.After using cyclopamine,the content of Cr,BUN and organ coefficient change reduced significantly, so did the pathological.Western blot detected kidney tissue Gli1 protein in each group.Compared to control group,there was no significant difference in Cyclopamine group.AA model group and AA+Cyclopamine group Gli1 protein expression was significantly higher compared with Cyclopamine Gli1 protein group,the difference was statistically significant.AA model group and AA Cyclopamine group Gli1 protein expression was significantly higher,the difference was statistically significant.Compared to the AA model group,AA+Cyclopamine group Gli1 protein levels have decreased significantly,the difference was statistically signifi-cant.Immunohistochemical assay detection found that kidney tissue proinflammatory cytokines TNF-α,IFN-γ,IL-6 expression and semi-quantitative score changed.Compared with the control group, TNF-α, IFN-γexpression of AA model group were significantly increased.After using Cyclopamine,the expression of TNF-α,IFN-γin kidney of AA model reduced significantly.IL-6 expression in AA model group was significantly higher than the control group.Conclusion:Cyclopamine AA can relieve arthritis and kidney injury in rats with arthritis AA,the Hh pathway was on activity state in the process,may altered expression of inflammatory factors.

Objective To investigate the expression of E-cadherin(E-cad),N-cadherin(N-cad),β-catenin (β-cat),which are the markers of Epithelial-mesenchymal transition and analyze their relationships with the clinicopathological features and the prognosis of the laryngeal squamous cell carcinoma.Methods The expression levels of E-cad,N-cad,β-cat in 76 tumor tissues and their corresponding adjacent normal laryngeal tissues were determined by immunohistochemistry method.Relationships between the proteins' expression and clinicopathological features were analyzed.Survival curves were calculated using the Kaplan-Meier method.Differences in the survival rates were analyzed by the log-rank test among different expression groups.Cox's regression model was used to examine the independent predictor of the prognosis of the laryngeal cancer.Results The expression levels of E-cad,N-cad,β-cat in tumor tissues and adjacent normal tissues were statistically significant (P ＜ 0.001).The expression level of E-cad and β-cat in the laryngeal carcinoma was related to lymph node metastasis,clinical pathological stage and differentiation degree (P ＜ 0.05),while N-cad expression level was associated with clinical stage and differentiation degree (P＜0.05).The expression of E-cad was correlated with the expression of β-cat (P =0.001),and the expression of N-cad was correlated with β-cat (P =0.02),but the expression of E-cad was not correlated with N-cad.There were four subgroups of patterns of E-cad and N-cad expression:E-cad (+)/N-cad (-),E-cad (+) /N-cad (+),E-cad (-) /N-cad (-),E-cad (-) /N-cad (+).The expression level of each group was related to the clinical pathological stage and differentiation degree (P ＜ 0.05).The expression level of E-cad/β-cat was associated with lymph node metastasis,clinical pathological stage and differentiation degree (P ＜ 0.01).Log-rank analysis showed that the prognosis of negative and positive groups was statistically different (P ＜ 0.05),and the combined analysis showed that the prognosis of E-cad/N-cad or E-cad/β-cat group was significantly different (P ＜ 0.01).Cox's regression model analysis showed that the clinical stage and β-cat were independent predictors of the prognosis of laryngeal carcinoma.Conclusions The low expression of E-cad,high abnormal expression of N-cad and β-cat played an important role in the occurrence and development of laryngeal carcinoma.It can provide a reference for evaluating clinical prognosis.The clinical pathological stage and β-cat can be used as independent predictors for the prognosis of laryngeal carcinoma.

Objective To discuss the indications and complications of primary closure of bile duct incision in laparoscopic bile duct exploration and balloon dilatation catheter dilatation to treat the papillary stenosis and the intrahepatic bile duct stenosis. Methods A pospective study of 42 ptients of bile duct incision closure primary in laparoscopic bile duct exploration and balloon dilatation catheter dilatation, laparoscopic bile duct exploration and extraction of bile duct stones with choledochotomy was first adopted in order to clear the stones, then followed by the balloon dilatation catheter(explosive pressure reached 2020 kPa, used 505kPa) to dilate the papillary stenosis and the intrahepatic bile duct stenosis (CT-7542～ CT-75104) until the stenosis was released. Whether the primary closure of duct incision was selected or not, it was based on the situation of intraoperative choledochoscopic exploration, if it had been selected, the closure of bile duct incision would accepted by using absorbable suture 4-0 or 5-0, without placing bile duct drainage.It was routinely to place the drainage tube in the oriffice of the lesser omentum. Results 41 out of 42 patients had obtained successful duct clearance, the dilatation of the stenosis to reach the expected expansion and without bile leakage. One patient had bile leakage about 30-150 ml daily persisted for 4 days through cured conservatively. Conclusion Eventually it was safe and effective for some patients who had completed successful duct clearance and the dilatation of the stenosis to reach the expected expansion with the balloon dilatation catheter. They were adopted to the primary closure of duct incision using absorbable suture and did not need to place bile duct drainage.

ObjectiveTo summarize the experience on laparoscopic common bile duct exploration(LCDE). MethodsFrom Mar. 1992 to Mar. 2003, there were 693 cases undergoing LCDE. ResultsThe procedure was successful in 373 out of 391 cases with primary closure of duct incision and 272 out of 302 cases of T tube placement. Membrane stenosis dilation was successful in 68 out of 74 cases. Eleven out of 15 cases of malignant stenosis was successfully treated by biliary endoprosthesis and stent without bile leakage. Seven cases were shifted to open CBD exploration. Bile leakage was cured conservatively in 24 cases. Endoscopy failed to totally remove residual stones in 11 cases. Four cases died postoperatively. Conclusion LCDE was safe and effective in the hands of skilled endoscopists.