Metformin activates chaperone-mediated autophagy and improves disease pathologies in an Alzheimer disease mouse model

Chaperone-mediated autophagy (CMA) is a lysosome-dependent selective degradation pathway implicated in the pathogenesis of cancer and neurodegenerative diseases. However, the mechanisms that regulate CMA are not fully understood. Here, using unbiased drug screening approaches, we discover Metformin, a drug that is commonly the first medication prescribed for type 2 diabetes, can induce CMA. We delineate the mechanism of CMA induction by Metformin to be via activation of TAK1-IKKα/β signaling that leads to phosphorylation of Ser85 of the key mediator of CMA, Hsc70, and its activation. Notably, we find that amyloid-beta precursor protein (APP) is a CMA substrate and that it binds to Hsc70 in an IKKα/β-dependent manner. The inhibition of CMA-mediated degradation of APP enhances its cytotoxicity. Importantly, we find that in the APP/PS1 mouse model of Alzheimer's disease (AD), activation of CMA by Hsc70 overexpression or Metformin potently reduces the accumulated brain Aβ plaque levels and reverses the molecular and behavioral AD phenotypes. Our study elucidates a novel mechanism of CMA regulation via Metformin-TAK1-IKKα/β-Hsc70 signaling and suggests Metformin as a new activator of CMA for diseases, such as AD, where such therapeutic intervention could be beneficial.

Establishment of a rapid identification of adverse drug reaction program in R language implementation based on monitoring data / 浙江大学学报·医学版

OBJECTIVE@#To establish a clinically applicable model of rapid identification of adverse drug reaction program (RiADP) for risk management and decision-making of clinical drug use.@*METHODS@#Based on the theory of disproportion analysis, frequency method and Bayes method, a clinically applicable RiADP model in R language background was established, and the parameters of the model were interpreted by MedDRA coding. Based on the actual monitoring data of FDA, the model was validated by the assessing hepatotoxicity of lopinavir/ritonavir (LPV/r).@*RESULTS@#The established RiADP model included four parameters: standard value of adverse drug reaction signal information, empirical Bayesian geometric mean value, ratio of reporting ratio and number of adverse drug reaction cases. Through the application of R language parameter package "phViD", the model parameters could be output quickly. After being encoded by MedDRA, it was converted into clinical terms to form a clinical interpretation report of adverse drug reactions. In addition, the evaluation results of LPV/r hepatotoxicity by the model were matched with the results reported in latest literature, which also proved the reliability of the model results.@*CONCLUSIONS@#In this study, a rapid identification method of adverse reactions based on post marketing drug monitoring data was established in R language environment, which is capable of sending rapid warning of adverse reactions of target drugs in public health emergencies, and providing intuitive evidence for risk management and decision-making of clinical drugs.

Construction of data remote monitoring and auditing system for clinical trials / 浙江大学学报·医学版

Clinical trial management system is independently developed by our hospital, which basically realized the whole process management and data collection of clinical trials. Based on the platform, the functional architecture of data remote monitoring and auditing was established. By desensitizing and encrypting of data, the project and subject hologram were visualized to facilitate to review of data. The data remote monitoring and auditing cloud platform adopts the B/S architecture pattern. Users register to apply for an account through the cloud platform, and access to the account via HTTPS security protocol. The authorized users were able to view the relevant items online to ensure the secure data transmission and easy operating. The electronic management of data is the direction of future efforts. By compliance with laws and regulations, the remote monitoring/auditing can be realized, and the data security and personal privacy can be ensured with the application of information technology. In this paper, the feasibility of remote monitoring/auditing mode is explored, specific technical schemes and system functions are suggested, and the realization scenarios are conceived in case of major public health emergencies.

Guideline Formulation of the Off-label Use of Drugs Based on Evidences / 中国药房

OBJECTIVE:To explore an effective method to formulate management-related strategies for off-lable use of drugs by the evidence-based medicine. METHODS:The process of guideline formulation included seven procedures,i.g. establishment ofguideliesformulation workgroup;investigation and selection of the status quo on off-label drug use;identification of the clinical problems;retrieval and evaluation and comprehensing of evidence;applification of GRADE in evidence quality grading;formation of the recommendations consensus;peer review and result publication. And eventually guidelines were formed based on the steps. This study took off-label use of rheumatoid immunoprotective subjects as a case to explore. RESULTS & CONCLUSIONS:Based on the evidence evaluation system and above 7 steps,the methods and process of guideline formulation on off-label use of rheuma-toid immunoprotective subjects that integrated administration,law,clinical medicine,pharmacy subjects were made .The process of guideline formulation fully reflects multidisciplinary characteristics of the workgroup,the advanced nature of the process,the comprehensiveness of evidence ,the rigor of evidence quality grading,and the normalization of consensus. It provides reference in methodology for establishing a comprehensive evidence-based evaluation and management system of off-label use of drugs for all clinical specialist disease. Therefore,this scientific research results may promote the standardization and legalization of the off-label use of drugs management in China.

Current status and future prospects of the development of clinical pharmacy in China: a SWOT analysis

In many industrialized countries, clinical pharmacy has developed into a separate discipline and become a vital part of inpatient care in hospitals. However, as compared to many established branches of medicine, clinical pharmacy is still in its infancy, with much room for growth, improvement, and recognition by both the medical community and patients. In this study, a widely-recognized development strategy analysis tool, Strength, Weakness, Opportunity and Threat [SWOT], was used to systematically address several key issues to the development of clinical pharmacy in China. This analysis aims to provide feasible recommendations for the development of clinical pharmacy in China by identifying current problems and growth opportunities. Full development of clinical pharmacy as a mature clinical discipline will help promote the rational use of drugs by both clinicians and patients and lead to enhanced drug efficacy and safety

Effect of Gemcitabine on viability and apoptosis of non-small cell lung cancer HCC827 in vitro / 中国免疫学杂志

Objective:To observe the effect of Gemcitabine ( GEM) on the viability and apoptosis of non-small cell lung cancer HCC827 in vitro. Methods:The cell viability,apoptosis and cell cycle of HCC827 cells induced by Gemcitabine were detected with cell counting kit-8 assay (CCK-8),Annexin V-FITC/PI staining and flow cytometry. The expression of Bcl-2 protein of cells treated with GEM was examined by Western blot assay. Results: There was significant inhibition effect on HCC827 cells treated with 0. 1-1 000 ng/ml of GEM,which can promote the occurrence of HCC827 cell apoptosis and arrest cell in the S phrase. The apoptosis induced by GEM was accompanied with the down regulation of Bcl-2 protein. Conclusion: GEM can inhibit the cell viability and induce the HCC827 cell apoptosis and S phrase arrest. Its cell dead type was apoptosis,which was related with the expression of Bcl-2 protein.

The regulatory effects of HTLV-1 Tax protein on HMGB1 gene in T cells / 中华微生物学和免疫学杂志

Objective To explore the regulatory effects of HTLV-1 ( human T-cell leukemia virus type 1 ) Tax protein on the expression of HMGB 1 ( high mobility group box 1 ) gene in T cells .Methods Total RNA and protein were extracted from Tax +-T cells ( TaxP ) , Tax--T cells ( TaxN ) and Jurkat cells which were stably transfected with pCMV-Tax and pCMV-Neo, respectively.Then, the expression levels of HMGB1 mRNA and protein in different CD 4+T cells were analyzed by real-time PCR and Western blot (WB).By using liposome-mediated method, pGL3-HMGB1-luc reporter genes and pGL3-neo-luc were tran-siently transfected into TaxP and TaxN cells and the basal transcriptional activity was observed in different T cells.Additionally, pCMV-Tax and pGL3-HMGB1-luc reporter genes were also co-transfected into Jurkat cells and the regulatory effects of Tax protein on HMGB 1 gene was detected .The chromatin immunoprecipi-tation (ChIP) assay was used to identify HMGB1 genomic sites directly targeted by Tax .Results The ex-pression levels of HMGB1 mRNA and protein in Tax+-T cells ( TaxP) were higher than those in Tax--T cells (TaxN).The transcription regulation trends for HMGB1 gene in TaxN and TaxP cells were similar but not identical in diverse T cells.pHLuc3 (containing -504-+83 HMGB1) showed the highest transcriptional ac-tivity of HMGB1 gene in both TaxP and TaxN cells , but HMGB1 transcriptional activity of pHLuc 6 in TaxP cells was significantly stronger than that in TaxN cells .Luciferase assays also showed that Tax protein promo-ted the transcription of HMGB1 gene in a dose-dependent manner .The ChIP assay further confirmed that Tax protein enriched at the HMGB1 region of -1163--1043.Conclusion The region of nt -504--383 is essen-tial for the basal promoter activity of -1163-+83 HMGB1 gene originated from pHLuc 6 reporter plasmid , and Tax protein enriched probably at the HMGB 1 site of -1163--1043 enhances HMGB1 transcription.

Changes of whole hemocyte parameters and hemorrheologic indexes after treatment in patients with cerebral infarction / 中国组织工程研究

BACKGROUND: The abnormal changes of hemorrheologic indexes are closely correlated with vascular injury, disorder of microcirculation, thrombosis and its development.OBJECTIVE: To investigate the changes of 25 indexes of hemorheology and whole hemocytes in patients with acute cerebral infarction.DESIGN: A controlled case study.SETTING: Department of Medical Laboratory, Xinxiang Medical College.PARTICIPANTS: Cerebral infarction group: Totally 127 patients with the first attack of acute cerebral infarction were selected from the Department of Neurology, the Second Affiliated Hospital of Xinxiang Medical College from January to December in 2002. The onsetof disease ranged from 4 to 12 hours, and the lesions were confirmed by cranial CT, the diagnostic standard accorded with the diagnostic guidelines for various cerebrovascular diseases formulated by the Fourth National Academic Conference for Cerebrovascular Diseases. Normal control group: Fifty-three healthy physical examinees with normal blood pressure and without diseases of heart,brain and kidney, as well as diabetes mellitus. There were no sigmficant differences in ages and male to femaleratios between the two groups.METHODS: ① Patients in the cerebral infarction group received fibrinogen reduction therapy, they were injected with two pieces of 10 u defibrin on the day of admission, and injected with one piece of 10 u defibrin again on the 3rd, 5th and 7th days respectively. ② Fasting venous blood (5 mL) was drawn with thromboliquine anticoagulated vacuum blood collecting tube,and 2 mL blood was coagulated with EDTA-K2. The whole blood viscosity,whole blood high-sheafing reduction viscosity, whole blood low-shearing reduction viscosity, whole blood high-sheafing relative index, whole blood lowsheafing relative index, equation K value of erythrocyte sedimentation, plasma viscosity, hematocrit, erythrocyte sedimentation, aggregation index of red blood cells (RBC), rigidity index of RBC, deformation index of RBC, counts of while blood cells (WBC) and RBC, mean volume of RBC, mean corpuscular volume (MCV), RBC distribution width (RDW), content of hemoglobin,mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet count, mean platelet volume (MPV) and platelet distribution width (PDW) were detected when the shear rate was 200 s-1, 30 s-1, 5 s-1 and 1 s-1 respectively. ③ The above indexes of the patients with cerebral infarction were detected within 24 hours after attack (exacerbation) and at two weeks after treatment (recovery period) respectively.MAIN OUTCOME MEASURES: The hemorrheologic and whole hemocyte indexes were compared between the two groups.RESULTS: All of the 127 cases in the cerebral infarction group entered the analysis of results at exacerbation, and 5 of them were not reexamined because of discharge and 122 cases entered the analysis of results at recovery period. All the 53 cases in the normal control group were involved in the analysis of results. ① Except RDW and MCHC, all the other indexes in the patients with cerebral infarction at exacerbation were significantly different from those in the normal control group (P＜0.05, 0.01). As compared with the normal control group, the whole blood low-shearing viscosity, whole blood low-shearing reduction viscosity, whole blood high-sheafing relative index, whole blood low-shearing relative index, plasma viscosity,hematocrit, aggregation index of RBC, WBC count and MCH all returned to normal in the patients with cerebral infarction at recovery period (P＞0.05).② As compared with the exacerbation, the RBC count and MPV were obviously decreased in the patients with cerebral infarction at recovery period (P ＜ 0.05), while the other indexes were similar (P ＞ 0.05).CONCLUSION: Abnormalities of hemorrheologic indexes commonly exist in patients with acute cerebral infarction, which indicates that the abnormalities of hemorrheologic and whole hemocyte parameters may be related to the occurrence of acute cerebral infarction.