Cross-talk of intracellular signaling pathways that share common components (hubs) is organized in form of a bow-tie network topology.Signaling cross-talk is functionally pleiotropic for target genes regulation, resulting in functional redundancy, synergism and antagonism, which should be precisely controlled to prevent signaling 'leaking' or 'spillover'.Thus, the biological system has evolved multiple insulating mechanisms to achieve stimulus-specific response that maintains intracellular homeostasis.The insulation mechanism of signaling cross-talk suggests: (1) the functional duality of cross-talk molecules that determine cell fate requires selectively targeting dysregulated cross-talk molecules while protecting the normal ones from off-target or unintended effects, and we propose them as the targetable cross-talk molecules;(2) cross-talk molecules are usually carried on the macromolecular complex as their functional platforms, thus the structural plasticity of conformational changes at the interaction surface of cross-talk molecules asks for intensive work on the relationship study between drug binding and biological activity, which we propose as the accessible cross-talk molecules.Therefore, signaling cross-talk and its insulation mechanism play instructive leading roles in resolving the bottlenecks of current drug R&D and improve the clinical outcome.

This study aimed at investigating the effects of the extract of Chinese herb,Nansheteng (C.orbiculatus Thunb.),on human HepG2 cells through the overexpression of mTOR.The GV238-mTOR recombinant plasmids were transfected into HepG2 cells using molecular biological technology.The expression level of mTOR was evaluated by means of relative activity of luciferase and western blot.Human hepatic carcinoma HepG2/mTOR++ cells were treated with C.orbiculatus extract in different concentrations (20,40,80,160 and 320 tg·mL-1) for 24 h.The mTOR protein expression was detected by western blot.It was found that the protein expression of mTOR in transfected HepG2 cells was significantly enhanced.C.orbiculatus extract significantly inhibited the proliferation of HepG2/mTOR++ cells.Simultaneously,C.orbiculatus extract inhibited mTOR at its protein level in a dose-dependent manner.In conclusion,we successfully constructed recombinant mTOR cloning vectors,and established the stable HepG2 cell line with the overexpression of mTOR.Besides,C.orbiculatus extract significantly inhibited mTOR protein expression in human hepatic carcinoma HepG2 cells.

This study was aimed to investigate the effect of Nan-She-Teng (Celastrus orbiculatus) extract on epithe-lial-mesenchymal transition(EMT) in HepG2 cells. Except the control group, human hepatocellular carcinoma HepG2 cells in other groups were treated with Celastrus Orbiculatus extract in different concentrations (10, 20, 40, 80, and 160 μg·mL-1). The protein expression levels related to EMT were detected by western blotting. At 48 h after fertiliza-tion, the zebrafish embryos were randomly assigned to 7 groups as follows: untreated control group (E3 buffer), DMSO group (E3 buffer with 1% DMSO), and different dosages treatment of C.orbiculatus extract (10, 20, 40, 80, and 160μg·mL-1) for 24 h. The protein expressions of mTOR signaling pathways were detected by western blotting. The re-sults showed that compared with the control group, C.orbiculatus extract significantly increased E-cadherin protein expression. Simultaneously, C.orbiculatus extract inhibited vimentin and mTOR signaling pathways at protein levels. It was concluded that to a certain extent, C.orbiculatus extract prevented EMT in HepG2 cells by modulating the mTOR signaling pathway. Therefore, it suggested that mTOR can be chosen as a new therapeutic target for clinical treatment of hepatic carcinoma.

Spinocerebellar ataxia type 2 (SCA2) is an autosomal dominant degenerative disease. The main clinical manifestation of SCA2 is progressive cerebellar syndrome, but a wide range of extracerebellar and non-motor symptoms can be observed clinically. The non-motor symptoms and extra-cerebellar signs in SCA2 patients are reviewed to provide a better understanding in cognition, diagnosis and treatment of the disease.

Objective:To develop a questionnaire on knowledge, attitude and practice of Helicobacter pylori (Hp) infection among physical examination population, and to verify its reliability and validity.Methods:This study was a cross-sectional survey. Based on the theory of knowledge, attitude and practice, the first draft of the questionnaire on knowledge, attitude and practice of Hp infection in physical examination population was designed by means of retrospective literature research, qualitative interview, discussion in the research group. The structure and items of the questionnaire were consulted and revised by the expert consultation method (Delphi method). Through pre investigation of 186 physical examination personnel from May to June in 2021, the final version of the "knowledge, attitude and practice questionnaire on Helicobacter pylori infection among physical examination population" was formed after adjustment and verification of the reliability and validity of the questionnaire.Results:The questionnaire of knowledge, attitude and practice of Hp infection in physical examination population included 3 dimensions and 28 items, and the cumulative variance contribution rate was 56.271%. The content validity index of each item level of the questionnaire was 0.75-1.00, and the content validity index of the total questionnaire was 0.94. The Cronbach alpha of knowledge, attitude and practice dimensions in this questionnaire were 0.862, 0.901 and 0.798 respectively. The overall Cronbach alpha of the questionnaire was 0.890, and the half reliability was 0.698. The test-retest reliability of the questionnaire was 0.919, and the test-retest reliability of each dimension was 0.924, 0.917 and 0.845.Conclusions:The questionnaire has good reliability and validity, and can be used to measure the level of knowledge, attitude and practice of Hp infection in physical examination population.

Objective To investigate the risk factors of postoperative pancreatitis after endoscopic retrograde cholangiopancreatography(ERCP),establish a quantitative risk prediction model,and conduct external validation.Methods The clinical data of patients with ERCP were analyzed.Among them,603 ERCP patients From January 2017 to January 2021 were selected as the modeling group,and 205 ERCP patients from March 2021 to March 2022 were selected as the validation group.Pancreatitis was diagnosed according to Atlanta standards.There were 45 cases in the modeling group and 23 cases in the validation group developed pancreatitis after ERCP.Compare the clinical data and biochemical indicators of patients with and without pancreatitis in the modeling group,and screen for risk factors of pancreatitis through multivariate Logistic regression analysis.Then,establish a risk prediction model and validate it.Results Univariate analysis showed that there were statistically significant differences in age,history of gastectomy,calculus of common bile duct,papillary foramen nodule type,pancreatic wire channel,sphincterotomy,serum total bilirubin and albumin between the two groups(P＜0.05).Multivariate Logistic regression analysis showed that the history of gastrectomy(O(R) = 6.417,95%CI:1.900～21.675,P = 0.000),calculus of common bile duct(O(R) = 3.442,95%CI:1.496～7.917,P = 0.000),papillary foramen nodule type(O(R) = 2.447,95%CI:1.072～5.585,P = 0.018),pancreatic wire channel(O(R) = 3.673,95%CI:1.609～8.383,P = 0.000),sphincterotomy(O(R) = 1.758,95%CI:1.140～2.711,P = 0.004),elevated total bilirubin(O(R) = 1.415,95%CI:1.084～1.847,P = 0.008)and decreased albumin(O(R) = 1.239,95%CI:1.016～1.510,P = 0.010)were independent risk factors for post ERCP pancreatitis.Establish a risk prediction model Y =-1.023 + 1.859×(history of gastrectomy)+ 1.236×(calculus of common bile duct)+ 0.895×(papillary foramen nodule type)+ 1.301×(pancreatic wire channel)+ 0.564×(sphincterotomy)+ 0.347×(elevated total bilirubin)+ 0.214×(decreased albumin).The receiver operator characteristic curve(ROC curve)showed that the area under the curve(AUC)of the model predicting pancreatitis in the modeling and validation groups were 0.895 and 0.864,respectively.After assigning values to each variable in the model,it was divided into low risk(0～5 points),medium risk(5～10 points),and high risk(≥10 points).The actual incidence of high-risk pancreatitis in the modeling and validation groups was significantly higher than that of low-risk patients,and the actual incidence of high-risk pancreatitis was significantly higher than that of medium risk patients,with statistical significance(P＜0.05).Conclusion The history of gastrectomy,calculus of common bile duct,papillary foramen nodule type,pancreatic wire channel,sphincterotomy,elevated total bilirubin and decreased albumin were independent risk factors for post ERCP pancreatitis.We have developed a quantitative risk prediction model with good predictive efficacy for pancreatitis,which has important clinical application value.

Parkinson′s disease (PD) is a common neurodegenerative disease. Its main pathological feature is the progressive loss of substantia nigra and other catecholaminergic neurons containing neuromelanin (NM). Therefore, NM may be closely related to the pathogenesis of PD. The content of NM can be detected by NM magnetic sensitive sequence imaging, and then it can be applied to the study of the neuropathological mechanism of NM and PD. This paper focuses on the physiological significance of NM, its role in the pathogenesis of PD and the prospect of NM as a biomarker to assist diagnosis and disease monitoring of PD.

OBJECTIVE：To establish th e fingerp rint and c onduct cluster analysis of Hefu zhiyang decoction （HFZYD），and establish the method for content determination of 8 components，so as to provide reference for the quality control of HFZYD. METHODS：UPLC method was adopted. The determination was performed on ACQUITY UPLC BEH C 18 column with mobile phase consisted of acetonitrile- 0.1％ glacial acetic acid solution （gradient elution ）at the flow rate of 0.25 mL/min. The column temperature maintained at 30 ℃，and detection wavelength was 236 nm. The sample size was 5 μL. The fingerprint of 10 batches of HFZYD was established with Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition）. Compared with substance control ，the common peaks were identified. The similarity evaluation was conducted. SPSS 21.0 software was used to conduct cluster analysis of 10 batches of samples. The contents of 8 components such as gallic acid in 10 batches of samples were determined by above UPLC. RESULTS ：There were 34 common peaks in the UPLC fingerprint of 10 batches of HFZYD ， identified as peak 1 was gallic acid ，peak 10 was ferulic acid ，peak 13 was astilbin ，peak 23 was paeonol ，peak 28 was dictamnine，peak 31 was obacunone ，peak 32 was fraxinellone ，and peak 34 was osthole. Its similarity with the control fingerprint was 0.923-0.979. Among 10 batches of samples ，S1，S2，S5，S6，S7，S8 and S 10 were clustered into one category ，and S 3，S4 and S 9 were clustered into one category. The average contents of above 8 components were 0.596-0.714，0.262-0.321，7.647-9.859， 0.113-0.644，0.170-0.202，0.854-1.281，0.631-0.857，3.243-3.548 mg/g，respectively. CONCLUSIONS ：UPLC fingerprint and the method for content determination of 8 components in HFZYD are established successfully.

ObjectiveTo understand the use of antibiotics in inpatients and the incidence and trend of hospital infections， to explore the implementation effect of comprehensive management measures， and to provide reference for hospitals to use antibiotics reasonably. MethodsBased on the hospital infection monitoring and management system， a retrospective analysis and comparison were conducted on the use of antibiotics， submission of microbial test samples， and incidence of hospital infections among inpatients in a tertiary hospital from 2012 to 2021. ResultsFrom 2012 to 2021， the use of antibiotics showed a downward trend， from 50.82% in 2012 to 41.29% in 2021. At the same time， the use rate of restricted and special antibiotics had also decreased， and the use rate of restricted and special antibiotics in patients without hospital infection was significantly lower than that in patients with hospital infection， and the microbial testing rate was also on the rise. The annual incidence rate of hospital infection was 0.69%‒1.92%， and the annual case-time prevalence rate was 0.79%‒2.17%. The annual average rate of the above two in 10 years was 1.18% and 1.34%， respectively. The results of the exponential smoothing model also showed that the utilization rate of antibiotics was decreasing and the incidence of nosocomial infection was stable. ConclusionLarge general hospitals took comprehensive management measures to strengthen the management of rational use of antibiotics， which led to a decline in the use rate of antibacterial drugs for inpatients and an increase in the rate of microbial examination. At the same time， the overall incidence of hospital infection was relatively stable， suggesting that the comprehensive management measures of antibacterial drugs in hospitals had achieved certain results. The current measures need to be optimized in the future to continuously improve the management level of rational use of antibacterial drugs.