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ObjectiveTo predict the mechanism of Sinitang in treating myocardial ischemia-reperfusion injury （MI/RI） based on network pharmacology and verify the prediction results by cellular experiments. MethodThe traditional Chinese medicine system pharmacology database and analysis platform （TCMSP） was employed for retrieval of the main components and potential targets of Sinitang. Online Mendelian Inheritance in Man （OMIM） and GeneCards were employed to obtain the targets of Sinitang in treating MI/RI. STRING was employed to construct the protein-protein interaction （PPI） network， and DAVID to perform gene ontology （GO） annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. Finally， cellular experiments were carried out to verify the predicted anti-MI/RI mechanism of Sinitang. ResultA total of 105 active ingredients and 234 targets of Sinitang were screened out， among which 116 targets were predicted to be involved in the treatment of MI/RI. The GO annotation gave 587 entries， including 417 biological process entries， 101 cell component entries， and 69 molecular function entries. The KEGG analysis enriched 125 signaling pathways， involving vascular endothelial growth factor （VEGF）， phosphoinositide 3-kinase/protein kinase B （PI3K/Akt）， forkhead box transcription factor O （FoxO）， hypoxia-inducible factor-1 （HIF-1） apoptosis and other signaling pathways. The results of cell viability assay showed that Sinitang increased the survival rate of H9C2 cells damaged by hypoxia/reoxygenation （H/R）. Sinitang decreased the levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and creatine kinase-MB （CK-MB） in H9C2 cells damaged by H/R. The results of flow cytometry demonstrated that Sinitang decreased the apoptosis rate of H9C2 cells damaged by H/R. Western blot showed that Sinitang down-regulated the expression of Bcl-2 related X protein （Bax） and up-regulated that of B-cell lymphoma-2 （Bcl-2） in H/R-injured H9C2 cells. ConclusionSinitang treats MI/RI in a multi-target and multi-pathway manner， which involves the signaling pathways associated with apoptosis.
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Tripterygium glycosides tablets (TGT) have good immunosuppressive activity, but they can also significantly injure the liver and kidney and its mechanism is unclear. In this study, delayed-type hypersensitivity (DTH) Balb/c mouse were administrated with different doses of TGT. Then the changes of sphingolipids levels in live, kidney and plasma as well as the mRNA expression levels of their metabolic enzymes were studied by the integrated targeted sphingolipidomics and transcriptomics methods to reveal the mechanism of efficacy and toxicity of TGT. It was found that low dose of TGT could significantly decrease levels of total ceramide in the plasma, long chain sphingolipids and saturate sphingolipids in the liver and kidney, but increase them in the plasma, which were related to the efficacy mechanism of TGT. High dose of TGT can significantly increase levels of total ceramide, Cer(d18:1/18:0)-1-P, long chain sphingolipids and decrease saturation sphingolipids mechanism. TGT can also cause significant changes of mRNA expression levels of various sphingolipid metabolic enzymes in the liver and kidney, which were correspond to the changes of sphingolipid levels. The efficacy and toxicity of TGT were related to the regulation of these key enzyme expression levels. In conclusion, the efficacy and toxic mechanism of TGT were closely related to the sphingolipids metabolism. A variety of potential biomarkers were found and they can provide valuable information for the evaluation of the efficacy and toxicity of TGT.
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Objective To seek a operation method of quality control of biochemical tests in peacekeeping mission via perspective of evidence-based laboratory medicine .Methods Literatures related to quality control of biochemical tests in international peace-keeping were referred ,compared and discriminated .Combined with 5-year peacekeeping laboratory experience ,a set of feasible ap-proaches were worked out and a practice evaluation was performed in a task period .Results Under the supervision of the quality control system ,providing accurate and reliable test results to the clinical ,developing and implementing quality control of biochemical tests were economical ,practical and reliable ,and could guarantee the effectiveness of reports of biochemical test in peacekeeping medical institutions .Conclusion Evidence-based laboratory medicine is important for quality control of biochemical tests in peace-keeping medical institutions .
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<p><b>OBJECTIVES</b>To study the phenomena of hepatitis B virus (HBV) integration into the tissues of hilar cholangiocarcinoma (HCCA) and to identify the integration sites in the host genome.</p><p><b>METHODS</b>Ten fresh HCCA samples were collected from the tissues by surgical ablation, 1 normal hilar bile duct sample selected as control. Cellular DNA were extracted by Wizard SV Genomic DNA Purification System. PCR-derived assay (HBV-Alu-PCR) was employed to amplify the viral-host junctions which contain the HBV sequence and the adjacent cellular flanking sequences. The PCR products were purified and subjected to sequencing by ABI-3730XL Auto DNA Analyzer. The sequence analysis of viral-host junctions was performed by DNASIS MAX 3.0 bioinformatics software. The insertion sites between viral and cellular sequences were identified through homology comparison using NCBI BLAST and MapViewer search.</p><p><b>RESULTS</b>In 10 HCCA samples, 5 were demonstrated to have HBV integration fragments with total 6 inserted sites identified. Sequence analysis from viral-host junction showed that HBV X gene inserted into host genome at random distribution with truncated fragments. HBV integration recurrently targeted the unknown region in upstream of CXXC finger protein-1 (CpG-binding protein) gene (4 cases). p53 tumor suppressor gene was also found at the integration site.</p><p><b>CONCLUSIONS</b>There is high integration rate of HBV DNA into cellular genome of HCCA. HBV integration is found frequently into or close to cancer-related genes. The findings demonstrate that HBV infection might have association with the pathogenesis of HCCA.</p>
Subject(s)
Aged , Female , Humans , Male , Base Sequence , Bile Duct Neoplasms , Genetics , Virology , Cholangiocarcinoma , Genetics , Virology , DNA, Viral , Genetics , Hepatitis B , Virology , Hepatitis B virus , Genetics , Virus IntegrationABSTRACT
Objective To survey the status of nosocomial infection after total knee arthroplasty,analyze the risk factors of nosocomial infection and possible prevention measures.Methods Datas were collected retrospectively on 80pmients (80 knee joints) who were treated by total knee arthrophsly,the patients were divided into two groups,group A with nosocomial infection and group B without nosocomial infection.Statistic patient's age,basic diseases situation,preoperative hemoglobin content,serum albumin,operation time,blood transfusions,indwelling urethral catheter time,antibiotic treatment time of the two groups.And study the location,pathogenic bacteria and outcomes of the nosocomial infection patients.Results 10 patients occured nosocomial infection,the infected site in turn is urinary tract in 5 cases,respiratory tract 4 cases,skin infections in 1 case,the incidence of nosocomial infection is 12.5％.In noscomial infection group,patient's age,blood transfusions,operation time and postoperative indwelling urinary canal time significantly higher than no nosocomial infection group,anemia,hypoalbuminemia have relevance of nosocomial infection,there is no difference between the two groups in basic diseases situation.Conclusions The nosocomial infection after total knee arthroplasty caused by multiple factors,patient's age,hypoalbuminemia,anemia,operation time and indwelling urethral catheter time is closely related with nosocomial infection
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<p><b>OBJECTIVE</b>To investigate the effect and safety of the hook needle knife for the treatment of stenosing tenovaginitis of flexor digitorum.</p><p><b>METHODS</b>From September 2007 to September 2008, 60 outpatients with stenosing tenovaginitis of flexor digitorum were randomized divided into the treatment group and the control group, 30 cases in each group. Among the patients, 44 patients were female and 16 patients were male, aged from 34 to 69 years, averaged 56 years, the duration of disease ranged from 1 month to 1 year, averaged 3 months. All the patients were treated with hook needle knife and local-blocking respectively. The patients were followed up for 6 months, and the relief of moving-pain, tender-pain, stretching-pain and resist-ing--pain were observed respectively. All the patients were evaluated by the symptoms with numerical rating scale.</p><p><b>RESULTS</b>The relief of moving-pain, tender-pain, stretching-pain and resisting-pain in the treatment group were significantly better than those of the control group; and the therapeutic effects of treatment group were better than those of the control group.</p><p><b>CONCLUSION</b>The method for treating stenosing tenovaginitis of flexor digitorum with hook needle knife has advantages of definite effects, micro-invasion and safety.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Medicine, Chinese Traditional , Minimally Invasive Surgical Procedures , Methods , Tendon Entrapment , General SurgeryABSTRACT
Objective To investigate the distribution of pathogenic bacteria of vaginal discharge in bacterial vaginosis.Methods The results of bacterial culture and drug sensitive tests of vaginal discharge from patients with bacterial vaginosis were analyzed.Results The positive rate of bacteria culture of vaginal discharge was 79.3%(115/145).The dominant bacteria were staphylococcus epidermidis 27.0%(31/115),staphylococcus intermedius and staphylococcus aureus 13.0%(15/115),which were obviously higher than other germs.The drug sensitive tests showed that staphylococcus were relatively sensitive to vancomycin,fosfomycin,amikacin and rifampin.But the drug resistance to penicillin,tetracycline,erythromycin and oxacillin was the highest.Conclusion The kinds of pathogenic bacteria in vaginal discharge are various.The main bacterium is staphylococcus,and drug resistance is very severe.The isolation and drug sensitive test of pathogenic bacteria play an important role in diagnosis and treatment of gynecological disease.
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<p><b>OBJECTIVE</b>To study the effect of Hepatitis B virus X (HBx) gene transfection on expression of human telomerase reverse transcriptase (hTERT) mRNA in human bile duct carcinoma cell lines QBC939 and to elucidate the significance of cis-activation of hTERT mRNA by HBx gene on the carcinogenesis of bile duct.</p><p><b>METHODS</b>QBC939 were cultured in vitro and co-transfected with eukaryotic expression vector containing the HBx coding region and cloning vector containing enhanced green fluorescent protein (EGFP) coding sequence using liposome-mediated gene transduction technique. Thirty six hours after transfection, EGFP expression, the indicator of successful transfection in cells, was determined. Flow cytometry was applied to determine the transfection efficiency. Cells were harvested and total RNA was extracted with TRI(ZOL) Reagent. The expression of hTERT mRNA in QBC939 was assayed by Reverse Transcription Polymerase Chain Reaction. The expression of HBx protein in QBC939 was detected by immunocytochemistry staining and western blotting.</p><p><b>RESULTS</b>The transfection efficiency was 29.6% for both HBx expression vector and vector control group. The expression of hTERT mRNA was significantly increased when transfected with HBx expression vector than that transfected with OPTI-MEM medium and vector only. The expression of HBx protein could only be found in the cells when transfected with HBx expression vector by immunocytochemistry staining and western blotting.</p><p><b>CONCLUSION</b>HBx gene transfection may up-regulate the transcriptional expression of hTERT mRNA in bile duct carcinoma cells. The cis-activation of hTERT gene by HBx gene is primary mechanism for carcinogenesis of biliary epithelia after HBV infection.</p>
Subject(s)
Humans , Bile Duct Neoplasms , Genetics , Metabolism , Cell Line, Tumor , DNA-Binding Proteins , Metabolism , Gene Expression Regulation, Neoplastic , Genetic Vectors , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Telomerase , Metabolism , Trans-Activators , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of human telomerase reverse transcriptase (hTERT) protein and mRNA in bile duct carcinomas and the adjacent tissues and to elucidate its role in bile duct carcinogenesis.</p><p><b>METHODS</b>The expression of hTERT protein and hTERT mRNA in the formalin-fixed paraffin-embedded specimens of 71 cases of bile duct cancers and 39 cases of adjacent tissues was detected by streptavidin-peroxidase immunostaining and in situ hybridization. The correlation was analysed statistically between the expression of hTERT protein and mRNA and clinicopathological parameters bile duct carcinomas.</p><p><b>RESULTS</b>The positive rate of hTERT protein expression and mRNA expression in malignant specimens was 78.9% (56/71) and 67.6% (48/71), while that in the adjacent tissues was 35.9% (14/39) and 23.1% (9/39), respectively. All the positive signals were found in the hyperplastic biliary epithelia. No significant correlation was established between hTERT expression and clinicopathological parameters.</p><p><b>CONCLUSION</b>hTERT gene transcription and protein expression is most likely involved in the proliferation and malignant transformation of bile epithelia and the malignant progression of bile duct carcinomas. The detection of hTERT expression may serve elucidating the carcinogenesis of bile duct.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bile Duct Neoplasms , Pathology , DNA-Binding Proteins , Immunohistochemistry , RNA, Messenger , Telomerase , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of HBV X gene (HBx mRNA) in extrahepatic biliary tract carcinomas and the adjacent non-cancerous tissues, and to analyzed the relationship between HBV infection and incidence of biliary tract carcinomas, thereby to elucidate the possible role of HBx in the carcinogenesis of biliary tract.</p><p><b>METHODS</b>The plasmid pSPX46 was digested by appropriate restriction enzyme. HBx fragment was obtained through gel extraction kit. The digoxigenin-labeled DNA probes for HBx mRNA were prepared by a random prime technique. The expression of HBx mRNA was detected in formalin-fixed- paraffin-embedded specimens from 71 cases of biliary tract carcinomas and 39 specimens of non-cancerous tissues adjacent to cancer by in situ hybridization. The correlations between HBx mRNA expression and clinicopathological parameters were statistically analysed in 71 cases of biliary duct carcinomas.</p><p><b>RESULTS</b>Forty-three of 71 malignant specimens had detectable HBx mRNA expression with a positive rate being 61%. Only 7 of 39 specimens of non-cancerous tissues adjacent to cancer had weak HBx mRNA expression, with a positive rate being 18%, and all these positive signals were found in the hyperplastic biliary epithelium. No significant correlation was found between HBx mRNA expression and clinicopathological parameters, but a strong positive correlation was found between HBx mRNA and protein expression.</p><p><b>CONCLUSION</b>There is a high frequency of HBx mRNA expression in extrahepatic biliary tract carcinomas. HBV infection and its gene integration might play a role to certain extent in the development of biliary tract carcinomas.</p>