Coronary Restenosis ; Drug-Eluting Stents ; Humans

Endolymphatic Hydrops ; Hearing Loss, Sudden ; Humans

Atherosclerosis ; prevention & control ; Humans ; Vaccination

Purpose:To investigate the cyclooxygenase-2(COX-2) expression in gastric carcinoma and investigate the effects of COX-2 expression on P-glycoprotein. Methods:The expression of COX-2 and P-glycoprotein was examined by immunohistochemical staining in 48 cases of gastric carcinoma and 10 cases of normal gastric tissues. Results:There was no positive signal of COX-2 detected in normal gastric tissue. The positive expression rate of COX-2 was 60.4 %(29/48) in gastric carcinoma. The expression of COX-2 was correlated with TNM stage and lymph node metastasis (P

ObjectiveTo evaluate the expression of interleukin-1 β ( IL-1β) ; interlenkin-8 ( IL-8 ) ; tumor necrosis factor-α,(TNF-α) in induced sputum in different stages of stable chronic obstructive pulmonary disease (COPD) and heath smoking controls.Methods90 healthy smoking controls and 100 stable COPD patients were enrolled.The levels of IL-1β,IL-8 and TNF-α in induced sputum were detected by radioimmunoassay.ResultsThe levels of IL-1β,IL-8 and TNF-α in stable COPD patients were much higher than those in healthy smoking controls (P =0.000).There are an inverse correlation between FEV1 and levels of IL-1 β,IL-8 and TNF-α in induced sputum (r=-0.678,-0.752,-0.615,all P＜0.05).ConclusionIL-1β,IL-8 and TNF-α may play important roles in the mechanism of airway inflammation in stable COPD.The levels of IL-1 β,IL-8 and TNF-α in induced sputum may be a marke of severity of airway inflammation in stable COPD.

OBJECTIVETo investigate the relationship between the level of di-2-ethylhexyl phthalate (DEHP) and idiopathic oligoasthenospermia by measuring the content of DEHP in the semen samples of different subjects.

METHODSWe obtained semen samples from 100 infertile men with idiopathic oligoasthenospermia, 50 working all the year round in the plastic greenhouse (group A) and the other 50 constantly dining from plastic meal boxes (group B). We also enrolled 50 normal male volunteers as controls (group C). We conducted semen analyses using a computer-assisted sperm analyzer, measured the DEHP concentration by reversed-phase high-pressure liquid chromatography, and subjected the data to statistic processing by t-test and correlation analysis.

RESULTSThe mean concentrations of DEHP in the seminal plasma were (0.72 +/- 0.48), (0.71 +/- 0.49) and (0.21 +/- 0.18) mg/L in groups A, B and C, respectively, significantly higher in A and B than in C (both P < 0.05). The DEHP concentration was negatively correlated with sperm motility (P < 0.05).

CONCLUSIONThe DEHP level in the seminal plasma is higher in infertile men frequently exposed to plastic products than in normal males and excessive DEHP may be one of the important factors of idiopathic male infertility.

Adult ; Case-Control Studies ; Diethylhexyl Phthalate ; adverse effects ; Humans ; Male ; Oligospermia ; etiology ; Plastics ; adverse effects ; Semen ; chemistry

Objective To study the optimum decoloration technological conditions of crude polysaccharides from Cordyceps militaris Link. (PCM) by macroporous resin. Methods With the decolorization rate and retention rate of PCM as indexes, static adsorption factor experiment was used for the optimization of seven kinds of macroporous resin. And then the optimum conditions of decolorization of PCM were investigated by the static/dynamaic adsorption single-factor experiments. Results D941 macroporous resin is the best. The optimum decoloration technological conditions were as follows:initial concentration of the sample was 50 mg/mL, pH value was 5.5, operating temperature was 45℃, the flow rate of dynamaic adsorption was 2.5 BV/h, and the sample solution volume was 5 bed volume per hour. Under the optimal conditions, decolorization rate and retention rate of PCM were ( 93.9 ± 3.4) % and (91.7 ± 2.2) %, respectively. Conclusion The resin D941 is suitable for the decoloration of PCM. The optimized method is feasible and reliable, having the prospects for popularizing.

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

Objective To develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .

Objective To investigate the relationship between aggrecan and YKL-40 in knee articular cartilage of Sprague-Daw-lay(SD) rats with osteoarthritis (OA) .Methods Fifty-six healthy SD rats were randomly divided into 7 groups ,8 cases per group . The one side of knee joint was randomly selected for performing the anterior cruciate ligment transection (ACLT) and establishing the OA model .The rats in one group were randomly killed on the day of operation and at postoperative 0 ,2 ,4 ,8 ,12 ,16 ,20 weeks . The femoral condyle cartilage samples at different time periods in the operated side were collected for conducting safranin O /fast green staining and HE staining .Meanwhile ,the OA pathological grade was made out according to the modified Mankin scale .The expression of aggrecan and YKL-40 in the cartilage with different stages of OA were analyzed by the immunohistochemistry meth-od ,and the status of expression were measured by average optical density (AOD) .The correlation between aggrecan and YKL-40 was analyzed .Results With the aggravation of OA ,the expression of aggrecan was gradually reduced and the expression of YKL-40 was gradually increased .The differences during the early ,middle and late phases of OA had statistical significance (P<0 .05) . The expression of aggrecan was negatively correlated with the expression of YKL-40(P<0 .05) .Conclusion The level of aggrecan is gradually reduced with the aggravation of OA .Aggrecan is negatively correlated with the YKL-40 level ,which may reflect the dedifferentiation degree of joint chondrocyte to some extent .