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Objective To use in vitro experiments to verify the changes of proliferation, senescence and apoptosis of hepatocellular carcinoma cells after inhibiting the expression of NEK7, and to explore the related molecular mechanism. Methods Western blot and RT-PCR were used to detect the expression of NEK7 in hepatocellular carcinoma cells and THLE-2 cells. A viral vector was designed to inhibit the expression of NEK7 based on the gene sequence. After hepatocellular carcinoma cells were transfected, we observed the changes of proliferation activity, cell senescence, cell apoptosis and cell cycle in vitro. Western blot was used to detect the expression of cell cycle-related factors. Results Compared with THLE-2 cells, NEK7 was highly expressed in hepatocellular carcinoma cells. After inhibiting the expression of NEK7 with shRNA, the proliferation of hepatocellular carcinoma cells was inhibited, the proportions of cell senescence and apoptosis were increased, meanwhile, the cell number in stage S and G2/M was significantly reduced, the cell cycle progression was blocked, the expression levels of C-myc, c-Fos, cyclin D1 and cyclin E were inhibited, P16 and P27 expression were increased, and CDK2, CDK4 and CDK6 expression were not significantly changed. Conclusion After inhibiting the expression of NEK7, the proliferation ability of hepatocellular carcinoma cells is reduced, cell senescence is promoted and apoptosis is induced; meanwhile, the cell cycle progress is blocked.
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Metabolic regulation has been proven to play a critical role in T cell antitumor immunity. However, cholesterol metabolism as a key component of this regulation remains largely unexplored. Herein, we found that the low-density lipoprotein receptor (LDLR), which has been previously identified as a transporter for cholesterol, plays a pivotal role in regulating CD8
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Dengue virus (DENV) is the most widely transmitted arbovirus in the world. Due to the lack of diagnostic technology to quickly identify the virus serotypes in patients, severe dengue hemorrhagic fever cases caused by repeated infections remain high. To realize the rapid differential diagnosis of different serotypes of DENV infection by immunological methods, in this study, four DENV serotype NS1 proteins were expressed and purified in mammalian cells. Monoclonal antibodies (MAbs) against NS1 protein were obtained by hybridoma technology after immunizing BALB/c mice. Enzyme-linked immunosorbent assay, indirect immunofluorescence assay, dot blotting, and Western blotting were used to confirm the reactivity of MAbs to viral native NS1 and recombinant NS1 protein. These MAbs include not only the universal antibodies that recognize all DENV 1-4 serotype NS1, but also serotype-specific antibodies against DENV-1, DENV-2 and DENV-4. Double antibody sandwich ELISA was established based on these antibodies, which can be used to achieve rapid differential diagnosis of serotypes of DENV infection. Preparation of DENV serotype-specific MAbs and establishment of an ELISA technology for identifying DENV serotypes has laid the foundation for the rapid diagnosis of DENV clinical infection.
Subject(s)
Animals , Humans , Mice , Antibodies, Monoclonal , Antibodies, Viral/metabolism , Dengue/diagnosis , Dengue Virus/immunology , Enzyme-Linked Immunosorbent Assay , Mice, Inbred BALB C , Sensitivity and Specificity , Serogroup , Viral Nonstructural Proteins/immunologyABSTRACT
Objective:To investigate the serum expression of long non-coding RNA overlapping transcription content KCNQ1OT1 (KCNQ1) gene in patients with coronary artery disease (CAD) and its clinical significance. Methods:A total of 196 patients treated in our hospital were divided into 2 groups:CAD group and Control group, the patients were without CAD. n=98 in each group. Expression levels of serum KCNQ1OT1 and P53 were measured by quantitative RT-PCR;the relationship between KCNQ1OT1, P53 and clinical features in relevant patients were analyzed. Results:Compared with Control group, CAD group had increased expression of serum KCNQ1OT1 and decreased expression of P53, both P Conclusion:CAD patients had obviously increased serum level of KCNQ1OT1;KCNQ1OT1 was independently related to CAD occurrence.
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Objective To establish a mouse model by transplanting subcutaneously human pancreatic cancer tissue fragments.Methods Surgically resected pancreatic cancer tissue fragments from patients were transplanted into NOD/SCID mice subcutaneously,and then the growth of the tumor and transplanting it into the next generation were observed.The growth rate,HE staining and immunohistochemistry staining of Ki67 and VEGF were compared.Results We have obtained 13 cases ofpancreatic cancer tissues and 6 cases of biopsy specimens.In 5 cases transplantation was successful,in onemouse model passing to fourth generation,in 4 models to second generation.With the increase of generaions,tumor growth accelerated.HE staining showed later passage cells behavior in an identical manner as the primary cells.Immunohistochemistry staining showed that expressions of Ki67 and VEGF are increasing.Conclusions Through transplanting human pancreatic cancer tissue fragments directly,we have constructed mouse model of pancreatic cancer successfully.With the passage of subculture,the malignant degree and invasiveness may increase.
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OBJECTIVE:To explore the establishment of performance indicator system for evaluating national essential drug system in China.METHODS:Reference to the field of health policy performance evaluation research and practice at home and abroad.WHO indicators for evaluating national drug policies,and a qualitative study on the theory and approach of policy evaluation was conducted.RESULTS & CONCLUSION:To establish a China-specific performance evaluation indicator system,policy goals should be specified,and evaluation criterions and policy components should be also defined firstly.An indicator system framework of national essential drug system which based on WHO recommendatory indicators should be proposed accordingly.But detailed indicators should be established based on field research and experts counseling.
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OBJECTIVE:To provide references for the establishment of performance indicator system for evaluating National essential medicines system in China. METHODS:To analyze the theory and approach of policy evaluation and WHO indicators for evaluating national drug policies. A literature study on relevant information was conducted. RESULTS & CONCLUSION:China may refer to WHO framework for the evaluation index system,according to the target of various stages of essential drugs system,in the implementation of our system of essential medicines on the basis of a full investigation,combined with our medical and health background and status,build essential drugs system of performance appraisal system and performance indicators.
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This article reviews the recent achievements in parasitology including new diagnostic techniques,molecular mechanism of parasitic pathogenesis,drug resistance,antigenic variation,parasite genomics and proteomics. The perspective development in the area is also discussed.