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OBJECTIVE@#To investigate the effects of lncRNA HOTAIR on the proliferation, invasion and migration of lymphoma cells through target gene miR-20a-5p and its molecular mechanism.@*METHODS@#After synthesizing HOTAIR siRNA and siRNA NC plasmids, they were transfected into lymphoma Raji cells, respectively. The expression of HOTAIR mRNA was detected by RT-qPCR. The proliferation, invasion and migration of lymphoma Raji cells were detected by CCK-8 assay, Transwell assay and cell scratch healing assay, respectively. The target gene of lncRNA HOTAIR was predicted by miRcode software, and the relationship between HOTAIR and target gene was analyzed by dual luciferase assay. After synthesis of miR-20a-5p inhibitor and inhibitor NC, Raji cells were transiently transfected. The expression of miR-20a-5p was detected by RT-qPCR, and the effects of down-regulation of miR-20a-5p on the proliferation, invasion and migration of Raji cells were analyzed. The overexpression plasmid of lncRNA HOTAIR and miR-20a-5p mimics were transfected into Raji cells simultaneously to analyze the proliferation, invasion and migration ability of Raji cells. After overexpression or down-regulation of miR-20a-5p, the expression of JAK/STAT3 signaling pathway related proteins was analyzed.@*RESULTS@#HOTAIR expression in Raji cells was decreased after transfection of HOTAIR siRNA (P<0.01), and miR-20a-5p expression was also decreased after transfection of miR-20a-5p inhibitor (P<0.01). HOTAIR had a targeting and negative regulation relationship with miR-20a-5p (r=-0.826). Silencing HOTAIR promoted the expression of miR-20a-5p and inhibited the proliferation, invasion and migration of Raji cells. Down-regulation of miR-20a-5p expression promoted the proliferation, invasion and migration of Raji cells. Effect of HOTAIR overexpression on the proliferation, invasion and migration of Raji cells could be reversed by up-regulation of miR-20a-5p. Down-regulation of miR-20a-5p expression activated the intracellular JAK/STAT3 signaling pathway.@*CONCLUSION@#HOTAIR affects the proliferation, invasion and migration of lymphoma cells by targeting miR-20a-5p, and its mechanism may be related to the activation of JAK/STAT3 signaling pathway.
Subject(s)
Humans , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation , Gene Expression Regulation, Neoplastic , Lymphoma , MicroRNAs/genetics , RNA, Long Noncoding/genetics , RNA, Small InterferingABSTRACT
Objective To improve ultrasonic unit configuration of the field medical team.Methods The problems of the ultrasonic unit were analyzed in operating platform setup,personnel and facility allocation,application of interventional ultrasound,cross infection prevention and etc,and some countermeasures were put forward accordingly.Results There were interdependence and interaction among the elements of the ultrasonic unit,and the rational configuration made the requirements satisfied for interventional therapy,massive casualty diagnosis,bed side application and etc.Conclusion Improved configuration enhances the efficacy of the ultrasonic unit of the field medical team.
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Objective To improve ultrasonic unit configuration of the field medical team.Methods The problems of the ultrasonic unit were analyzed in operating platform setup,personnel and facility allocation,application of interventional ultrasound,cross infection prevention and etc,and some countermeasures were put forward accordingly.Results There were interdependence and interaction among the elements of the ultrasonic unit,and the rational configuration made the requirements satisfied for interventional therapy,massive casualty diagnosis,bed side application and etc.Conclusion Improved configuration enhances the efficacy of the ultrasonic unit of the field medical team.
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ObjectiveToinvestigatetheplasmidgenechangesinquinolone‐sensitivePseudomonasaeruginosa.Methods 31iso‐lates from January 2011 to December 2013 from various qualified clinical samples in the hospital were collected .In the 31 isolateds , 16 isolates proved sensitive to quinolones by using K‐B method were used as research objects in the study .The isolates growing ou‐side the sensitive ring of ciprofloxacin paper were selected to continuously transferred into other culture dishes until the resistance to quinolones were acquired .Plasmid transformation and extraction were performed on those isolates to confirm the existence of drug‐resistanceplasmidsacquired,andthroughPCRandgenesequenceanalysistodeterminethetypeofplasmids.Results 2iso‐lates of quinolone‐sensitive Pseudomonas aeruginosa acquired drug‐resistance plasmids qnrS and were resistant quinolones induced by continuous transferring for 9 times .Conclusion If antibiotics of inhibitory concentration were often used for the treatment of Pseudomonas aeruginosa infection ,drug‐resistance plasmids were acquired easily .
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To investigate whether there is mutation in DC-SIGN promoter region in patients with chronic hepatitis B (CHB) and healthy persons previously infected with hepatitis B virus (HBV) and to explore the relationship between the mutation in dendritic cell-specific intercellular adhension molecule-3-grabbing nonintegrin (DC-SIGN) promoter region and HBV.Methods The studied population was composed of two cohorts:47 CHB patients and 20 healthy persons previously infected with HBV.The mutation in DC-SIGN promoter region was detected with PCR,single-stranded conformational polymorphism and heteroduplex analysis,cloning,sequencing and aligning the published DC-SIGN promoter sequence.Results The characteristic mutation within DCSIGN promoter region in HBV infected individuals was observed.In the DC-SIGN promoter region,4 hot spot mutations located in positions - 139,- 142,- 222,and - 336 were observed in the CHB patients,but only 1 spot mutation located in position - 139 was observed in the healthy persons previously infected with HBV.The -336C which was absent in the healthy persons previously infected with HBV was shown in 11 CHB patients (23.40％).The - 139T was far more frequent in the healthy persons previously infected with HBV ( 100％ ) than in the CHB patients (34.04％).Conclusion In the DC-SIGN promoter region,-336C may be a genetic risk factor for developing CHB,but -139T may be associated with protection against HBV.
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<p><b>OBJECTIVE</b>To investigate whether there were particular HBx gene mutations associated with hepatocellular carcinoma (HCC) development in patients.</p><p><b>METHODS</b>The HBx genes were examined in 51 paraffin-embedded tumor tissue samples from patients with HCC and 25 serum samples from HBV carriers from southern China by nested polymerase chain reaction (PCR), single-stranded conformational polymorphism analysis, heteroduplex analysis and DNA sequencing. The HBx genes with deletion variations (HBx-d382, HBx-d431) from tumor tissues were cloned and transfected into QSG7701 cells. Then, the biological characteristics of the transfected cells were analyzed in nude mice by MTT assay, soft agar colony formation assay, flow cytometry and xenografting.</p><p><b>RESULTS</b>Deletion mutation and point mutation were found in the HBx genes of HCC tumor tissues, and there were some differences between the HBx gene mutations in genotype B and those in genotype C. More mutations were found in genotype C than those in genotype B (t=-2.522, P < 0.05), but the deletion variations (HBx-d382, HBx-d431) were detected in genotype B HBV from HCC liver tissues. The HBx genes with deletion variations (HBx-d382, HBx-d431) were recombinant with pcDNA3 and transfected into QSG7701 cell lines successfully, which established four permanent transfected QSG7701 cell lines, including pcDNA3/HBx-d382/QSG7701, pcDNA3/HBx-d431/QSG7701, pcDNA3/HBx/QSG7701, and pcDNA3/QSG7701. pcDNA3/HBx-d382/QSG7701 and pcDNA3/HBx-d431/QSG7701 grew faster and had more potential colony formative activity than those of pcDNA3/QSG7701. Moreover, pcDNA3/HBx-d382/QSG7701 and pcDNA3/HBx-431/QSG7701 cells inoculated in nude mice produced tumors more rapidly than those of pcDNA3/HBx/QSG7701, and pcDNA3/QSG7701. The volumes of the tumors in nude mice were also obviously larger in pcDNA3/HBx-d382 and pcDNA3/HBx-d431 groups than those in pcDNA3/HBx/QSG7701 and pcDNA3/QSG7701 groups.</p><p><b>CONCLUSION</b>Our results suggest that HBx gene mutations occur frequently in HCC tissues, and the deletion at nt382-400 of the HBx gene might play a role in carcinogenesis of HCC in southern China.</p>
Subject(s)
Adult , Aged , Animals , Female , Humans , Male , Mice , Middle Aged , Base Sequence , Carcinoma, Hepatocellular , Virology , Cell Line, Tumor , DNA, Viral , Genetics , Hepatitis B Surface Antigens , Blood , Hepatitis B virus , Genetics , Liver Neoplasms , Virology , Mice, Inbred BALB C , Mice, Nude , Point Mutation , Polymorphism, Single-Stranded Conformational , Sequence Analysis, DNA , Sequence Deletion , Trans-Activators , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the relationship between serum high-sensitivity C-reaction protein (hs-CRP) and heart fatty acid binding protein (h-FABP) on cardiac accidents in patients with unstable angina pectoris (UAP).</p><p><b>METHOD</b>Serum levels of hs-CRP, h-FABP, cardiac troponin-I (cTn-I) and creatine kinase MB isoenzyme (CK-MB) were measured and cardiac accidents within 2 weeks after the test were observed in 74 patients (male 45) with stable AP (SAP) and 56 patients (male 29) with UAP.</p><p><b>RESULTS</b>The incidence of cardiac accidents was significantly higher in UAP group (26.8%) than that in SAP group (10.53%, P < 0.001). Serum hs-CRP [(7.64 +/- 2.18) mg/L vs. (1.78 +/- 0.62) mg/L, P < 0.001], h-FABP [(16.46 +/- 5.28) microg/L vs. (3.15 +/- 2.61) microg/L, P < 0.001] and cTn-I [(1.28 +/- 0.43) microg/L vs. (0.67 +/- 0.09) microg/L, P < 0.001] levels were also significantly higher in UAP group than those in SAP group. The serum hs-CRP and h-FABP levels for patients with cardiac accidents in the SAP group were (6.32 +/- 2.06) microg/L and (8.76 +/- 3.83) microg/L respectively, which were higher than those for the patients having no cardiac accidents in the control (P < 0.01). The serum hs-CRP, h-FABP, cTn-I and CK-MB levels in patients with cardiac accidents were significantly higher than those in patients without cardiac accidents in both SAP and UAP groups.</p><p><b>CONCLUSION</b>Measuring traditional parameters for myocardial damage (cTn-I and CK-MB) in combination with hs-CRP and h-FABP is valuable for predicting the risk of recent cardiac accidents for AP patients.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Angina, Unstable , Blood , Diagnostic Imaging , C-Reactive Protein , Metabolism , Case-Control Studies , Coronary Angiography , Fatty Acid Binding Protein 3 , Fatty Acid-Binding Proteins , BloodABSTRACT
OBJECTIVE:To investigate the anti-HBV activities of deoxynojirimycin(DNJ)derivatives N—benzyl—1—DNJ(P-DNJ)and N—nonyl—1—DNJ(NN-DNJ)in vitro.METHODS:Human hepatoma carcinoma cell lines HepG2 2,2,15,which were transfected from HBV DNA,were taken as target cells,cells were cultured with different concentrations of test drugs, with HBsAg,HBeAg and HBV DNA in the cultured supernatant determined by time-resolved immunofluorometric assay and fluorescent quantitation PCR assay on day 6th and 10th day,meanwhile,with the cytotoxicity of DNJ derivatives determined by MTT assay.RESULTS:No cytotoxic effects was noted when the test concentration of P-DNJ and NN-DNJ was within 5～125?g?mL-1,HBsAg and HBeAg levels and HBV DNA secretion decreased significantly at a concentration of 125?g?mL-1.CONCLUSION:P-DNJ and NN-DNJ showed anti-HBV activity in the in vitro cell culture experiment.