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Objective To establish a quantitative detection method for Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA detection system with double internal standards(IMC-PCR-ELISA) .Methods The immunomagnetic (Dynabeads? ) which could specifically capture Mycobacterium tuberculosis were prepared .According to Mtp40 and IS6110 gene sequence of Mycobacterium Tuberculosis ,2 pairs of primers(upstream primer was modified with Biotin at 5′end) ,2 same-length mutant fragments with PCR amplified fragments ,and 3 capture probes(modified with digoxigenin at 3′end) were designed .Myco-bacterium tuberculosis were captured by immunomagnetic ,then detected by PCR-ELISA with double internal standards .Results The IMC-PCR-ELISA method could yield quantitative results in about 4 h with a detection limit at 5 × 103 copies/mL .There was a fine linear relationship between the copies of Mtp40(IS6110)in fact and in the calculation through formula when the concentrations of low internal standards were 30-70 copies/mL and the concentrations of high internal standards were 8 000-12 000 copies/mL (r2 =0 .998) .No nonspecific amplification was observed .Conclusion A rapid and quantitative method for the detection of Myco-bacterium tuberculosis was established successfully .The IMC-PCR-ELISA method was rapid ,sensitive ,secific and quantitative .
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Objective To investigate the effect of methylprednisolone pretreatment on cardiopulmonary bypass(CPB)-induced intestinal barrier injury in patients undergoing cardiac surgery.Methods Ninety NYHA Ⅰor Ⅱ patients,aged 30-50 yr,weighing 50-75 kg,scheduled for elective cardiac surgery with CPB,were randomly divided into 3 gnoups(n =30 each):control group without CPB(group Ⅰ),control group with CPB(group Ⅱ)and administration of methylprednisolone before CPB group(group Ⅲ).Anesthesia was induced with midszolam,fentanyl,etomidate and rocuronium and maintained with intravenous infusion of propofol and intermittent iv boluses of fentanyl and rocuronium.The patients were mechanically ventilated after tracheal intubation.In group Ⅲ,methylprednisolone 10 mg/kg was injected intravenously before operation and CPB.While in groups Ⅰ and Ⅱ,the equal volume of normal saline was injected instead.The blood samples were taken from the central vein before induetion of anesthesia(T1),before CPB(T2),at 30 min after the beginning of CPB(T3),at 30 rin afier the end of CPB(T4)and at 120 min after operation(T5)for determination of the plasma endotoxin concentration.Infection was recorded within 7 days after operation.Results The plasma endotoxin concentrations at T1 were within the normal range in all groups,without significant difference among the three gnoups(P ＞0.05).The plasma endotoxin concentration at T3-5 and incidence of postoperative infection in group Ⅲ were significantly lower than those in group Ⅱ,while higher than those in group Ⅰ(P ＜ 0.05).Conclusion Methylprednisolone pretreatment can reduce CPB-induced impairment of the intestinal harrier function in patients undergoing cardiac surgery.
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High mobility group A2 protein (HMGA2), an architectural factor, is highly expressed in various cancer types including lung cancers. It is a candidate target for cancer therapy. RNAi is an effective gene silencing method with low cost and less time-consuming. It is possible to exploit this technology in therapy. Here, 5 siRNAs targeting Hmga2 gene (HMGA2 siRNA1-5) were designed and synthesized. MTT assay, colony formation assay, transwell assay and flow cytometry were used to evaluate the effects of these siRNAs on lung cancer cell lines (NCI-H446 and A549). Results from cell proliferation, clone formation, migration and apoptosis showed that HMGA2 siRNA1, 3, 5 could affect these aspects for both lung cancer cell lines. Among the five siRNAs, HMGA2 siRNA5 showed the greatest inhibition effects. The inhibition effects of HMGA2 siRNA5 are sequence specific and are not due to the induction of interferon response. Taken together, siRNAs targeting Hmga2 gene are potential candidates for lung cancer gene therapy.
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Objective:To study the relation between clinical index and prognosis and the level of vascular endothelial cell growth factor(VEGF) in serum of non small cell lung cancer(NSCLC) patients before and after radiotherapy.Methods:VEGF serum of 39 patients with NSCLC and in 20 healthy individuals as control were analyzed with double sandwich in assay ELISA.Results:The level of VEGF serum elevated significantly before radiotherapy in NSCLC patients group as compared with the control(P0 05).And the level gradually declined 0,2～3 months after radiotherapy (P
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Objective To examine if high-dose glucocorticoid has any cerebral protective effects aftercardiopulmonary resuscitation. Methods Twenty-one healthy mongrel dogs of both sexes weighing 10 .5-15 kg weresubjected to cerebral ischemia using Pittsburgh standard ventricular fibrillation-cardiac arrest (VF-CA) model. Theanimals were anesthetized with intraperitoneal pentobarbital sodium 30 mg?kg~(-1) and mechanically ventilated thertracheal intubation (V_T 15 ml?kg~(-1), RR 18-20 bpm). Right femoral vein was cannulated for fluid administration(lactated Ringer's solution 0. 15 ml?kg~(-1)?min~(-1)) and right femoral artery was cannulated for BP monitoring. ECGwas continuously monitored. The animals were randomly divided into 3 groups: group A received only routineresuscitation treatment (control, n = 5); group B received dexamethasone 5 mg?kg~(-1) (n = 8) and group Cmethylprednisolone 30 mg?kg~(-1) (n = 8) after successful cardiopulmonary resuscitation. 8 hours after resuscitationthe animals were anesthetized and three pieces of brain tissue were obtained from right parietal lobe for light andelectron microscopic examination and determination of dopamine (DA) and 5-hydroxy-tryptamine (5-HT) contentsusing high-performance liquid chromatography- electrochemical technique (HPLC- ECD ). Results The DA and 5 -HT contents in parietal lobe of cortex were significantly decreased in group B and C as compared with group A (P