Objective To establish mixed-sandwich ELISA detection system by screening aptam-ers of MPT64 antibodies with SELEX to detect clinical serum samples, and explore the potential laboratory diagnosis value of this method. Methods To detect the affinity of the final round ssDNA library to MPT64 antibodies inhibited by MPT64 antigen with the competitive ELISA method, optimize the mixed-sandwich ELISA detection method that was aptamer-serum-horseradish peroxidase labeled goat anti-human IgG anti-body detection system to detect 230 cases of clinical serum samples as well as the lowest concentration of MPT64 antibodies and the linear range. Results In competitive ELISA test results, the percentage of inhi-bition effect of MPT64 antigen to final round ssDNA library is from 0.25% to 80% when the MPT64 antigen concentration rised from 2 μg/ml to 256 μg/ml. The Optimized detection system of mixed-sandwich ELISA was constitute of the concentration of ssDNA coated with 0.1μg/hole, serum dilution of 1/200, horseradish peroxidase labeled goat anti-human IgG antibody concentration of 1/40 000. The lowest concentration of MPT64 antibody is 3 mg/L and the linear range is between 10 mg/L and 1000 mg/L. The serum samples of 100 cases of tuberculosis patients, 100 healthy individuals and 30 cases of non-tuberculesis were tested in this system and the test result was analyzed with Graphpad Prism, the difference of tuberculosis group and healthy group was statistically significant (P<0.001 ), the difference of TB group and non-TB control group was also statistically significant (P<0.001). The specificity and the sensitivity was 96.1% and 31.0% re-spectively. Conclusion The aptamer mixed-sandwich ELISA method will play an important role in the sero-logical diagnosis of tuberculosis.

Objective To invesgate the influence of aceclofenac to cartilage cell proliferation and substrate metabolism in osteoarthritis.Methods Select 65 cases of patients with osteoarthritis who were treated in Department of Rheumatism and Hematology of Affiliated Hospital of Qinghai University from September 2011 to September 2013.Patients were divided into observation group(n=35)and control group(n=30).The observation group were treated with aceclofenac,while control group were treated with ibuprofen,Compare the efficacy of patients in two group.Compare human articular cartilage cells of 5-bromodeoxyuridine(BrdU)positive rate,the content of glycogen protein(PG)and collagen type Ⅱ in two groups. Results The effective rate of observation group and control group was 74.29%and 73.33% respectively.There was significant difference between the two groups. Compared with control group,the joints pain,joints tenderness,15 meters walking time,daily activity ability(P<0.05).The incidence of adverse reactions of observation group(1 1.43%)was significantly lower than that of control group(P<0.05 ).The difference of BrdU positive rate in two groups had no statistical significance. Ⅱ type collagen content of observation group was significantly higher than that of control group (P <0.05 ).PG of observation group was significantly lower than that of control group(P<0.05).Conclusion The efficacy of aceclofenac is good and the incidence of adverse reactions is low in treatment of osteoarthritis.Aceclofenac can alleviate clinical symptoms of patients by increasing collagen type Ⅱ and decreasing PG.

AIM:To study the effects and mechanism of peroxisome proliferator-activated receptors(PPARs)ligands,fenofibrate and pioglitazone,on ventricular remodeling in pressure overload rats.METHODS:A pressure overload model was established by the constriction of abdominal aorta in Wistar rats.The hemodynamics and ventricular remodeling parameters,plasma and myocardial renin activity,angiotensin Ⅱ and aldosteron,the mRNA expression of angiotensin Ⅱ type 1 receptor(AT1)were investigated in the constriction of abdominal aorta group(CAA group,n=7)at 12-week after operation and treated experimental groups in which rats were treated with fenofibrate(F group,n=8),pioglitazone(P group,n=7),concomitant fenofibrate and pioglitazone(F+P group,n=6)for 12 weeks since 2 days after operation.The sham-operated rats served as controls(n=8).RESULTS:The ratio of left ventricular weight to body weight,mean arterial pressure,left ventricular systolic pressure,left ventricular end diastolic pressure,left ventricular systolic pressure and heart rate were significantly lower,the maximum left ventricular pressure rising and declining rates(?dp/dtmax)were significantly higher in all treated experimental groups than those in CAA group.Fenofibrate or pioglitazone had no effect on plasma and myocardial levels of renin,angiotensin Ⅱand aldosteron.The mRNA expression of AT1 was downregulated in treated groups except F group.CONCLUSION:PPAR ligands have no effect on plasma and myocardial levels of renin,angiotensin Ⅱand aldosteron,but fenofibrate and pioglitazone inhibit ventricular remodeling,decrease preload and afterload,increase ?dp/dtmax in pressure overload rats.The expression of mRNA of AT1 is downregulated in myocardium of pressure overload rats by the PPAR? signaling pathway.

This article gives a profound exploration on the theoretical origin, prohibitions and mechanisms of moxibustion for heat syndrome. Based upon the ancient and modem literature, this article also gives a classified summarization on diseases of moxibustion for heat syndrome, in order to obtain a thorough understanding about the theory and clinical application of moxibustion for heat syndrome, hence to further perfect the theory of moxibustion and guide the clinical practice.

Objective To perform the bibliometric research on the literatures of the endoscopic retrograde cholangio pancreatog-raphy(ERCP) in the last ten years to provide reference for accurately grasping the international research situation and selecting the cutting-edge technological topics .Methods The literatures of ERCP in the database Web of Science from 2001 to 2011 were collect-ed .The CiteSpace software was adopted to analyze the distribution characteristics ,key point literature ,hotspots fields and research frontier of articles of ERCP .Results 2 921 citations of the research articles were retrieved .The analysis showed that America was strongest in the ERCP research field ;10 key point articles were detected .The high frequency keywords were complications ,manage-ment ,sphincterotomy ,etc .,the concentricity ranged 0 .06-0 .24 .The high frequency burst words were gold standard ,procedure-re-lated-complications ,biliary-stents ,but the concentricity was 0 .Conclusion The research hotspots of ERCP field are the occurrence rate of ERCP postoperative complications ,diagnosis and treatment ,but no obvious research cutting-edge is formed .

ObjectiveTo evaluate the clinical efficacy and safety of nylon ligation combined with endoscopic high-frequency electric snare for the treatment of colorectal polyps.MethodsThe clinical data of 345 patients with colorectal polyps who received the treatment of nylon ligation combined with endoscopic high-frequency electric snare at the Zhongshan Hospital of Fudan University from January 2006 to January 2011 were retrospectively analyzed.Complications including bleeding and perforation were observed.Postoperative wound healing and local residual recurrence were assessed during follow-up.ResultsA total of 362 colorectal polyps were successfully removed.The mean diameter of the colorectal polyps was 2.7 cm (range,1.5-4.0 cm). Operative bleeding occurred in 1 patient,and perforation in 1 patient.The results of histological examination showed that 93.6％(339/362) colorectal polyps were adenoma,17 were hyperplastic polyps,5 were inflammatory polyps,and caneeration of colorectal polyps in 1 patient was detected.Endoscopic follow-up was completed in 334 patients at the first month after operation,and 9 patients with residual benign colorectal polyps received endoscopic resection.No residue or recurrence was found in the other patients.Endoscopic follow-up was completed in 308 patients at the second months after operation,and no residue or recurrence was found.The overall effective rate was 97.1％ (299/308).ConclusionEndoscopic nylon ligation combined with endoscopic high-frequency electric snare is effective and safe for the treatment of colorectal polyps.

Objective To evaluate the clinical efficacy and safety of metallic hemoclips in the treatment of gastric defects during endoscopic full-thickness resection. Methods Patients with submucosal tumors derived from the muscularis propria and adhesion of the gastric serosa diagnosed by EUS and CT were enrolled in the study.A total of 62 patients,22 males and 40 females,mean age 58.5 years,were recruited into the present study from June 2009 to December 2010,in which 37 patients were with tumors in gastric fundus,20 in gastric body and 5 in antrum.All patients were treated with endoscopic full-thickness resection.After the operation,metallic hemoclips were used to close the defects through endoscopic biopsy channel.The closure success rate with metallic hemoclips were assessed.The patients were followed up endoscopically for evaluation of efficacy and safety of the procedures.Results All patients underwent endoscopic full-thickness resection.Uncontrollable bleeding occurred in 1 patient,who subsequently underwent emergent laparoscopic surgery and was excluded from the study.Metallic hemoclips were applied to close defects in 57 patients,and metallic hemoclips combined with omentum obstruction in 4 patients.The overall success rate was 100％ in 61 patients.Endoscopic follow-up was completed in all patients in average of 4.4 months.The clinical wound healing rate was 100％.No complications such as fever,abdominal pain were found in 2months after the procedures.Conclusion Application of metallic hemoclips during endoscopic full-thickness resection for gastric defects is a safe and effective technique.

To explore and compare the response of the protease inhibitors or non-nucleoside reverse transcriptase inhibitors-based therapeutic impact on metabolic indices in hepatitis C virus (HCV)/human immunodeficiency virus(HIV) co-infected patients.A randomized,open,and control approach was performed to enroll 273 cases of HCV/HIV co-infected patients on their initial visits and to choose protease inhibitors(PIs group) or non-nucleoside reverse transcriptase inhibitors (NNRTIs group) based therapy treatments for one year.Laboratory results of metabolic indices before and after the treatment were collected.After treatment,the levels of triglyceride in NNRTIs and Pls groups were (1.93 ± 0.99) mmol/L and (1.62 ± 0.93) mmol/L respectively,high density lipoprotein-cholesterol were(1.28 ± 0.55) mmol/L and (1.08 ± 0.53) mmol/L,low density lipoprotein-cholesterol were (2.60 ± 1.44) mmol/L and (2.22 ± 1.16) mmol/L,fasting plasma glucose were (5.92 ± 1.21) mmol/L and (4.79 ± 0.47) mmol/L,serum creatinine were (70.5 ± 14.6) μmol/L and (56.6 ± 8.3) μmol/L,and serum amylase were(66.9 ± 27.5) U/L and(62.7 ± 33.8) U/L respectively.The difference between the two groups was statistically significant(all P＜0.01).There is a therapeutic impact on metabolic indices in patients wtih HCV / HIV co-infection after non-nucleoside reverse transcriptase inhibitors-based regimen.

To study the protective effect of astragalus saponin extracts (AS) on kidneys of diabetic rats. Totally 32 diabetic rats induced by streptozotocin (STZ) were divided into AS high and low dose groups, the positive control group and the model group (DM group) and orally administered with 50 mg x- kg(-1) x d(-1) AS 200, 25 mg x kg(-1) x d(-1) valsartan, 10 mL x kg(-1) x d(1) physiological saline, respectively. Another 8 healthy rats were collected in the normal control group (NC group, physiological saline 10 mL x kg(-1). d(-1)). All rats were treated for consecutively 6 weeks. After the administration, the body weight was measured every week, the concentration of blood glucose was monitored on week 2, 4 and 6. The total urine and total urinary protein (U-TP) in 24 h were measured by the metabolic cage method on week 6; At the end of week 6, blood samples were collected from hearts to detect blood urea nitrogen (BUN), serum creatinine (Scr), uric acid (UA) , total cholesterol (CH) triglyceride (TG) by biochemical methods. Kidneys were collect to calculate the kidney hypertrophy index and observe the pathological sections. The laboratory results show that in the DM group, the blood glucose, metabolic cost in 24 h, kidney hypertrophy index, U-TP, BUN, Scr, UA, TG were significantly higher than that in the NC group (P < 0.01, P < 0.05) , with significant pathological changes; After the intervention with AS, the metabolic value in 24 h, kidney hypertrophy index, U-TP, BUN, Scr, UA, TG were significantly lower in the high dose group (P < 0.01, P < 0.05), and the kidney hypertrophy index, BUN, Scr, UA, TG in the low dose group were also significantly lower (P < 0.05), with slight reduction in renal pathological changes in both groups. In conclusion, Astragalus saponin extracts have a certain protective effect on kidneys of diabetic rats.
Animals ; Astragalus Plant ; chemistry ; Blood Glucose ; metabolism ; Blood Urea Nitrogen ; Diabetic Nephropathies ; metabolism ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Saponins ; administration & dosage ; Uric Acid ; metabolism

Objective To construct a new recombinant immunotoxin expression vector by using human VEGF165 and a truncated pseudomonas exotoxin A ramification (PE38) gene, and explore the expression of the VEGF165-PE38 fusion protein in HEK293 cells. Methods VEGF165 was cloned by polymerase chain reaction (PCR). PE38 gene was gained from an vector plasmid pRB391 by restriction endonuclease digestion, and then inserted to the eukaryotic expression vector pIRES2-EGFP. After the eukaryotic recombinant vector pIRES2-VEGF165-PE38-EGFP was identified by restriction endonuclease digestion and sequence analysis, the vector was transfected into HEK293 cells by liposome protocol. RT-PCR and ELISA method was used to confirm the expression of the fusion gene in the HEK293 cells. Results Restriction endonuclease digestion and sequence analysis revealed the VEGF165-PE38 fusion gene was cloned into the eukaryotic expression plasmid vector pIRES2-EGFP successfully. The pIRES2-VEGF165-PE38-EGFP fusion gene could express in the HEK293 cells. Conclusion The result provide the basis for search of the targeted cytotoxic activity to tumor vascular endothelial cells and may have some potential value in clinical application.