Objective To establish a new non radioimmunoassay for detecting insulin. Methods We put together the techniques of chemiluminescent analysis, flow injection analysis and immunoassay and established a method,flow injection chemiluminescent immunoassay, for insulin determination using sandwhich procedure with the horseradish peroxdase (HRP) as label of antibody and using the detection system of synergistic enhanced chemiluminescence of para phenylpenol and sodium tetraphenylborate with HRP catalysed oxidation of luminol. Results Satisfactory results were obtained in the dynamic range 0.15～60.00 ?U with a relative coefficient of 0 997 2 ( P

Objective To study the relationship between the operative programs and curative effects in treating non-reflex pubovesical muscle neurogenic bladder by transurethral division of internal sphincter. Methods Bilateral or one laternal division of internal sphincter were performed separately,followed-up for 3 months to 5 years in postoperation,and analysed data obtained .Results Bilateral division had a much higher improving rate(92%) than that of one laternal(60%)(P

Objective To observe the effect of Chinese medicine of replenishing the liver and kidney on the concentration of 5-hydroxytryptamine(5-HT)in hippocampus and the expression of tryptophan hydroxylase 2 (TPH2)in median raphe nuclei in perimenopausal depression model rats and to explore its mechanism.Methods The animal model was established by resecting the ovaries of female rats,then giving isolation-housing in combination with 21 days of chronic unexpected mild stress(CUMS).High performance liquid chromatogram-electrochemical detection(HPLC-ECD)were employed to measure 5-HT in hippocampus,the hemi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)and fluorescence immunohistochemistry were employed to detect the relative expression of TPH2 mRNA and number of TPH2 immunoreactive neurons in median raphe nuclei.Results The concentration of 5-HT in hippocampus was increased in rats of the medicine group compared with that in the model group((2543.06±859.59)ng/g tissue,(1845.81±233.55)ng/g tissue,F=9.617,P＞0.05).RT-PCR showed that the relative expression of TPH2 mRNA in medicine group was much higher than that in model group ((1.282±0.158),(0.985±0.120),F=3.552,P＜0.05).Immunofluorescence showed that the number of TPH2 immunoreaetive neurons in raphe nuclei had statistical significance between medicine group and model group ((152.74±68.52),(74.12±38.01),F=7.939,P＜0.01).Conclusion Chinese medicine replenishing liver and kidney have effects of improving perimenopausal depression,increasing the expression of TPH2 in median raphe nuclei,which leads to enhance the synthesis of 5-HT in hippocampus might be one of its mechanisms.

Objective To investigate the effect of atorvastatin calcium on serum endothelin-1(ET-1), polyamine oxidase(PAO), heart-type fatty acid binding protein(H-FABP), vascular endothelial growth factor(VEGF), 100β, inflammatory cytokines and nerve function in patients with acute cerebral infarction.Methods According to the random number table, 113 patients were randomly divided into two groups (n=61) and control group (n=52).The control group received conventional treatment methods, and observation group received atorvastatin calcium on the basis of control group. The treatment course was two weeks.Serum ET-1, PAO, H-FABP, VEGF, S100β, inflammatory cytokines and NIHSS score were compared between two groups before treatment, 7d and 14d after treatment.Results The serum levels of ET-1, PAO, H-FABP after 7d, 14d treatment of observation group was significantly lower than that of control group, respectively (P<0.05).The VEGF level of observation group after 7 d, 14 d treatment of observation group was significantly higher than that of control group, respectively (P<0.05).The S100βlevel after 7 d, 14 d treatment of observation group was significantly lower than that of control group, respectively (P<0.05).The hs-CRP, IL-8, TNF-αlevels after 7 d, 14 d treatment of observation group was significantly lower than that of control group, respectively (P<0.05).NIHSS score after treatment of observation group was significantly lower than that of control group (P<0.05).Conclusion The atorvastatin calcium can improve neurological function in patients with brain injury through reducing serum ET-1, PAO, H-FABP and S100βlevels, promote angiogenesis through increasing VEGF expression, and alleviate inflammation and ischemia-reperfusion injury through reducing inflammatory cytokines, thereby promote neurological functional recovery.

Objective To explore the effects of combining tumor necrosis factor-α (TNF-αt) inhibitor with adenosine A2b receptor antagonist CVT-6883 on asthmatic lung irflammation in mice.Methods A total of 40 female Balb/c mice were evenly randomized into 5 groups,including normal control group,asthma group,CVT-6883 group,CVT-6883 + etanercept group,and etanercept group.The pathological changes in the lungs were determined and the number of white blood cells(WBC) and eosinophil(EOS) in the bronchoalveolar lavage fluid(BALF) was counted by cell count in each group.The levels of TNF-α in BALF were evaluated by enzyme-linked immunosorbent assay (ELISA).The expression of adenosine A2b receptor mRNA in the lung tissues were measured by reverse transcriptionpolymerase chain reaction (RT-PCR).Results 1.The lung tissue in asthma group,dyed by HE,was found to have a large number of airway inflammatory cell infiltration,thickening of the bronchial mucosa,the alveolar septa widened and fracture.In the CVT-6883,CVT-6883 + etanercept and etanercept group,the pathological changes were relieved.2.The WBC and EOS counts in BALF of the asthma group[(413.8 ±5.8)/L,(139.3 ± 1.4)/L] were higher than those of the normal control group [(24.0 ± 1.3)/L,(1.8 ± 0.1)/L,P ＜ 0.05].The WBC and EOS counts of the CVT-6883 group[(111.5 ±3.8)/L,(3.3 ±0.1)/L],the etanercept group + the CVT-6883 group[(173.8 ±3.9)/L,(10.4 ± 0.2)/L],and the etanercept group[(138.4 ± 3.0)/L,(4.1 ± 0.1)/L] were lower than those of the asthma group (P ＜0.05).3.Compared with the control group(100.4 ± 5.7) ng/L,the TNF-α concentration of the asthma group (145.2 ± 8.8) ng/L was significantly higher (P ＜ 0.05) ; the TNF-α concentration of CVT-6883 group (130.9 ± 5.9) ng/L,CVT-6883 + etanercept group(115.7 ± 8.2) ng/L and the etanercept group(122.0 ± 8.7) ng/L,were significantly decreased compared with asthma group (P ＜ 0.05).4.In asthma group (8.9 ± 1.1) compared with the control group(0.6 ± 0.2),the A2bAR (adenosine A2b receptor) mRNA expression was upregulated (P ＜ 0.05) ; CVT-6883 group(1.6 ±0,3),CVT-6883 + etanercept group(2.5 ±0.6) and the etanercept group(5.3 ±0.4),the A2bAR(adenosine A2b receptor) mRNA expression was significantly decreased compared with asthma group (all P ＜0,05).Conclusion Combination of TNF-α inhibitor with adenosine A2b receptor antagonist can reduce asthmatic lung inflammation.

Objective To discuss the feasibility and long-term effect of free latissimus dorsi muscle flap in repair of severe lower extremity injury in children. Methods From July 1999 to June 2004, nine child patients (at age of 6-13 years) with severe lower extremity injury involving soft tissue defects a-round the calf and the foot associated with complex open fractures, bare dislocation, and injury of the nerve, tendon and artery were repaired with free latissimus donsi flap, with flap area ranging from 30 cm ×12 cm to 10 cm × 5 cm. Results All the latissimus dorsi flaps survived, with success rate of 100%. A follow-up for 4-9 years showed that the flap had sound shape and function and normal blood supply, without significant influence on donor area. Conclusion Latissimus dorsi flap has advantages of constant anatomical site, abundant blood supply, massive area, strong anti-infection ability and less in-fluence on donor area and hence is an ideal method for repairing severe lower extremity injury in children.

Objective To study the effect of postoperative anticoagulant therapy after splenectomy in patients with cirrhosis and portal hypertension.Methods One hundred and forty patients with cirrhosis and portal hypertension receiving splenectomy and periesophagastric devascularization were divided into anticoagulant group (76 cases) and control group (64 cases) by random number table,patients in anticoagulant group received postoperative anticoagulant therapy,principally according to the platelet count,gave ligustrazine,aspirin,low molecular heparin after operation; patients in control group without postoperative anticoagulant therapy.Postoperative monitoring platelet count and D-dimer,ultrasound or CT check the presence of portal vein thrombosis.Results Platelet count,D-dimer levels in anticoagulant group and control group in 2 days after operation were significantly increased,the difference was significant compared with preoperative [anticoagulant group:(95.73 ± 28.06) × 109/L vs.(38.41 ± 11.96) × 109/L,(3.61 ± 0.18) mg/L vs.(0.42 ± 0.09) mg/L;control group:(92.56 ± 27.75) × 109/L vs.(35.13 ± 11.38) × 109/L,(3.26 ± 0.16) mg/L vs.(0.37 ± 0.09) mg/L,P ＜ 0.05].Platelet count and D-dimer levels between two groups at preoperative and postoperative in 2 days had no statistical significance (P ＞ 0.05).Ten cases of control group occurred postoperative portal vein thrombosis,anticoagulant group were 3 cases,portal vein thrombosis incidence of anticoagulant group [3.95％ (3/76)] compared with control group [15.62％(10/64)] was statistically significant (P ＜ 0.05).Conclusion Postoperative anticoagulant therapy after splenectomy in patients with cirrhosis and portal hypertension is an effective method to prevent portal vein thrombosis.

[ ABSTRACT] AIM:To investigate the inhibitory effect of Am80 on neointima hyperplasia in carotid arteries after balloon injury and to observe the interaction between Krüppel-like factor 4 (KLF4) and retinoic acid receptorα(RARα). METHODS:Neointima hyperplasia in carotid arteries was observed by hemotoxylin and eosin staining.The expression of KLF4 and cyclin D1 was examined by immunostaining and Western blotting analysis.To detect the interaction between KLF4 and RARαin the vascular tissue, the injured arteries were harvested, and the protein extracts were prepared and subjected to co-immunoprecipitation assay.RESULTS:Compared with injured group, Am80 significantly reduced neointi-mal hyperplasia and the thickness ratio of intima to media.Am80 not only up-regulated KLF4 or RARαexpression in caro-tid arteries, but also increased the interaction between KLF4 and RARαat tissue levels.CONCLUSION:Am80 inhibits neointima hyperplasia in carotid arteries after balloon injury by promoting the interaction between KLF4 and RARα.

Objective To investigate the effect of PBL combined with EBM applied in the teaching of fever of unknown origin. Methods PBL combined with EBM teaching was applied in fever of unknown origin course for 30 clinical medicine specialty(eight years) students of Tongji class of grade 2009(experiment group), while PBL teaching was applied in fever of unknown origin course for 30 clinical medicine specialty (eight years) students of Tongji class of grade 2008 (control group). After teaching, the theory examination for both basic knowledge and case analysis was organized for all students of both groups. At the same time the questionnaire survey was conducted to 30 students of grade 2009 to evaluate the teaching effect. The results were assessed by using SPSS 18.0 statistical software for the T-test of the experimental group and the control group.Inspection level was α=0.05. Results The theory test score of students in the experimental group was (93.5±3.2) point, signifi-cantly higher than that of the students in the control group(84.7±2.8). There was statistically signifi-cant difference between the scores of the two groups of students (P=0.00). Survey results showed 19 students ( 63 . 33%) thought that the development of PBL teaching combined with evidence-based medicine teaching had its necessity, and 16 students(53.33%) thought that the teaching method im-proved their clinical thinking ability of logical reasoning. Conclusion The concept of PBL combined with EBM has achieved significant resultsinthe teaching offever of unknown origin, and it is necessary to carry out this teaching mode in medical colleges with certain teaching strength.

OBJECTIVETo investigate the therapeutic effect of retroviral endostatin gene transfer on the human colon cancer cell line, LoVo.

METHODSA retroviral vector pLESSN expressing secretable endostatin was constructed and packaged with a titer of 8.2 x 10(5) CFU/ml. A LoVo cell line was subjected to retrovirus-mediated endostatin gene transfer. The proviral integration of endostatin was analyzed with PCR. The function of endostatin was tested by MTT assay in vitro and a mouse xenograft model in vivo.

RESULTSAfter transfection and superinfection, amphotropic retrovirus was collected, and transduction with amphotropic retroviruses resulted in endostatin proviral integration. The endostatin secreted by transduced LoVo cells markedly inhibited cell growth up to 67% (P<0.001), compared with the control cells. The gene expression of endostatin in LoVo colon tumor cells significantly inhibited tumor growth in vivo. There was an 86% reduction in tumor size in the endostatin-transduced group, accompanied by a reduction in vessels, compared with the control group (P<0.01).

CONCLUSIONRetroviruses can allow functional expression of the endostatin gene in human colon tumors, showing promise for an antitumor strategy using antiangiogenesis.

Cell Division ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; therapy ; Endostatins ; genetics ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Retroviridae