Objective To study the effects of tri-ortho-cresyl phosphate(TOCP) on mitochondrial membrane permeability in hen's nerve tissues and investigate the mechanism of the organophosphorus ester-induced delayed neurotoxicity(OPIDN).Methods Adult Roman hens were randomly divided into four groups,including three treated groups and one control group(24 in each group).The hens in the experimental groups were treated with TOCP by gavage at the single dosages of 185,375 and 750 mg/kg respectively.TOCP was dissolved in corn oil and administered at 0.65 ml/kg.The control hens received an equivalent volume of corn oil by gavage.The hens were sacrificed on the 1st,5th,15th and 21st day after treatment and the cerebrum,cerebellum,spinal cord were dissected and homogenized in ice bath.The mitochondria in these nerve tissues were extracted to determine the changes of the membrane permeability and membrane potential.Results Compared with the control group,no significant increase of the mitochondrial membrane permeability in the cerebrum was observed in treated groups.In the cerebellum,the membrane permeability in the 185 mg/kg group had no significant changes,while in the 375 and 750 mg/kg groups it increased significantly on the 1st and 5th day after TOCP treatment(P

Objective To analyze the risk factors of Gram‐negative bacterial blood‐stream infection in ICU for conducting the risk evaluation and guiding medication .Methods The inpatients were diagnosed with Gram‐negative bacterial blood‐stream infec‐tion in ICU of the Renmin Hospital of Wuhan University from January 2013 to December 2014 were retrospectively surveyed and analyzed .The risk factors of blood‐stream infection caused by Gram‐negative bacteria were analyzed and screened by the Logistic re‐gression analysis method .Results A total of 172 case‐times of blood‐stream infection occurred in ICU during this period ,including 93 case‐times of Gram‐negative bacterial infection .The Gram‐negative pathogenic bacteria were Acinetobacter baumanii ,Klebsiella pneumoniae ,Acinetobacter calcoaceticus baumanii ,E .coli ,pseudomonas aeruginosa ,etc .Except E .coli was mainly originated from community acquired infection ,other bacteria were mainly originated from nosocomial infection .In order to differing from other pathogenic bacterial blood stream infection ,the Logistic regression analysis results showed that the independent risk factors of G‐negative bacterial blood‐stream infection in ICU had serum PCT levels over 10 .0 ng/mL (OR= 60 .52 ,P= 0 .001) ,receiving the therapy of carbapenem and third generation cephalosporins (OR=16 .09 ,P=0 .03) ,hospitalization duration less than 2 weeks be‐fore suffering from disease (OR=13 .79 ,P=0 .03) and digestive system basic disease(OR=12 .94 ,P=0 .01) .Conclusion Gram‐negative bacterial blood‐stream infection in ICU of the Renmin Hospital of Wuhan University is mainly caused by multi‐drug resist‐ant bacteria .Serum PCT level over 10 .0 ng/mL ,hospitalization duration less than 2 weeks before infection ,receiving the therapy of carbapenem and third generation cephalosporin and basic diseases of digestive system are the independent risk factors influencing the occurrence and diagnosis of blood‐stream infection .

AIM: To explore the effects of processing on components group of Semen Raphani. METHODS: By comparing HPLC maps of Semen Raphani samples prepared with different methods and computing similarity,(analysis) the change law of the components group. RESULTS: The mechanism of inhibiting the enzyme by processing Semen Raphani was found and C3 was able to produce new compounds A209,B221. CONCLUSION: Processing Semen Raphani can change the components group through influencing the decoction process,eventually produce different clinical effects.

Objective To study the influence of arachidonic acid (AA) on L-type calcium channel in rabbits sin-gle cardiomyocyte and its mechanism of antiarrhythmia. Method The single ventricular cardiomyocyte was isolat-ed by using enzyme dispersion method and whole-cell clamp-patch technique was used to record L-type calcium current.All data were analyzed using ANOVA. Results AA inhibited Ica-L in a concentration-dependent manner. The application of 3 μmol/L, 10 μmol/L and 20 μmol/L arachidonic acid reduced the density of peak Ica-L from (10.79±0.93)pA/pF to (8.99±0.43)pA/pF to (7.60±0.35)pA/pF and to (5.60±0.30)pA/pF, respctive-ly (n=7, P<0. O1 ). The Ica-Lpartially resumed after washout. The AA up-shifted the I-V curves of Ica-L without changes of their shape,peak and reverse potentials. The AA also markedly shifted the inactivation curve to left, and prolonged the recorvery time from inactivation,but did not change the curve of calcium channel activation. Con-clustions By acceleration of L-type calcium channel inactivation and prolongation of recorvery time from inactiva-fion,arachidonic acid can reduce the calcium ion influe and prolong effective refractory period, playing the role of antiarrhythmia.

BACKGROUND:Endothelial progenitor cel s have been shown to play an important role in the pathogenesis of traumatic diseases in recent years. OBJECTIVE:To explore the effect of magnetic labeled endothelial progenitor cel transplantation on renal function of diabetic rats through a MRI imaging study.METHODS:Sixty Wistar rats were randomly divided into normal (no treatment), control and experimental groups. Intraperitoneal injection of 40 mg/kg streptozotocin was performed to make a rat model of type 1 diabetes in the control and experimental groups. Four weeks after modeling, rats in the experimental group were given intravenous injection of magnetic labeled endothelial progenitor cel s (0.15 mL, 1×109/L). Fasting blood glucose, serum insulin, serum creatinine, urea nitrogen and 24-hour urinary protein levels in rats were measured at 8 weeks after cel transplantation. MRI was used to trace transplanted cel s in vivo in comparison with renal biopsy findings, and rat body mass and kidney weight were measured to calculate kidney weight index. RESULTS AND CONCLUSION:After modeling, fasting blood glucose, serum creatinine, urea nitrogen and 24-hour urinary protein levels as wel as kidney weight index were increased significantly (P<0.05), while the insulin level decreased (P<0.05). Compared with the model group, the endothelial progenitor cel transplantation reversed these indices (P<0.05). Additional y, in the experimental group, there was slightly longer T1 and shorter T2 signals as wel as marked lesion edge, and the FLASH sequence became more remarkable compared with the T2-weighted RARE sequence. The other groups showed no significant low signal changes. Magnetic-labeled positive cel s in the experimental group showed by the MRI were consistent with the tissue biopsy results, while no positive cel s were found in the model and normal groups. To conclude, the magnetic labeled endothelial progenitor cel transplantation can improve renal dysfunction in diabetic rats to a certain extent.

Aim To investigate the contribution of cardiac L-and L/T-type Ca 2+ channels blockers on matrix metalloproteinases (MMPs) protein expression and fibronectin (FN) degradation following myocardial infarction(MI). Methods Rat MI model was established by permanent ligation of left coronary artery. Infarcted rats were orally treated with placebo, amlodipine (L-channel blocker, 4 mg?kg -1?d -1) or mibefradil (L/T-Channel blocker, 10 mg?kg -1?d -1) for 7 days before induction of myocardial infarction. Protein levels of MMP-2,3,9 and FN distribution were assayed by immunoflorescence at 1, 3, 7 days post coronary occlusion in left ventricular free wall (LVFW) and myocardium interventricular septum (IS). Results Pathological changes of myocardial tissues in IS and LVFW showed typical myocardial remodeling. The hypertrophy was dominant in IS, but in LVFW the characteristics including disordered alignament, hypertrophy of the myocytes, the discontinuity, dissolving of carediomyofibrills, and the hyperplasia of interstitial tissue were found 7 days after MI. The protein levels of MMP-2,3,9 increased in IS 1,3,7 days post MI gradually, whereas the protein levels of FN decreased gradually, the protein levels of MMP-2,3,9 increased significantly in LVFW 7 days post MI, and the protein levels of FN decreased significantly compared to control group, respectively(P

AIM　The purpose is to construct D-hydantoinase genetic engineering strain for the purpose of the industrial production of D-p-hydroxyphenylglycine. METHODS　D-hydantoinase gene was created from Pseudomonas putida 9801 by PCR technique and inserted into pMD18-T vector. The recombinant plasmid was transformed into several Escherichia coli strains. The positive transformants with D-hydantoinase activity were obtained by the two step screening, digoxigenin DNA labeling in situ hybridization and D-hydantoinase activity assay. RESULTS　The D-hydantoinase activity of the genetic engineering strain E.coli BL21/pMD-dht was 1700 U*L-1 and increased as high as 8 times compared with those of wild-type strain Pseudomonas putida 9801. The subunit molecular weight of recombinant D-hydantoinase was about 53 kDa measured by SDS-PAGE. The amount of the recombinant D-hydantoinase was about 20 percent of total bacterial soluble proteins. CONCLUSION　The genetic engineering strain E.coli BL21/pMD-dht possesses the initial industrial production prospects.

BACKGROUND:Some studies indicate that PI3K/Akt signaling pathway is associated with the expression of glucose transporter 4 (GLUT4) and the function of cAMP response element binding protein (CREB) in skeletal muscle. However, it is stil unclear whether PI3K/Akt signaling pathway has the effects on CREB and GLUT4 in skeletal muscle of the rats with high-fat diet and treadmil exercise. OBJECTIVE:To investigate whether PI3K/Akt signaling pathway has the effects on CREB and GLUT4 in gastrocnemius muscle of the rats with high-fat diet and treadmil exercise. METHODS:A total of 70 rats were fed with normal diet for 2 weeks, and randomly divided into common feed group (n=20) and high-fat feed group (n=50). Rats in both groups were respectively fed with common feed and high-fat feed for 8 weeks. The rats in the common feed group were equaly assigned to common feed quiet group and common feed exercise group. 20 rats from the high-fat feed group whose body weight was 1.4 times of common rats were randomly and equaly assigned to obese quiet group and obese exercise group. Rats in the quiet groups did not do exercises. Rats in the exercise groups received adaptive sports for 1 week and medium-intensity treadmil exercise for 8 weeks. RESULTS AND CONCLUSION:(1) Impairments of PI3K/Akt signaling pathway appeared in obese rats, however, the quantity of GLUT4 expression did not change obviously in gastrocnemius muscles of obese rats. The reasons for the decrease of the nuclear protein CREB level of gastrocnemius muscles of obese rats might be related to the decrease of pAkt-Ser473 level. (2) The increase of the quantity of GLUT4 expression was accompanied by significantly up-regulated pAkt-Ser473 level by exercise intervention in gastrocnemius muscles of obese rats. Exercise intervention significantly increased the expression of nuclear protein CREB in gastrocnemius muscles of chow-fed rats and obese rats, which was consistent with the changes of pAkt-Ser473. These findings suggest that pAkt-Ser473 can play an important role in the effects of high-fat diet and exercise intervention on GLUT4 and CREB protein expression in gastrocnemius muscles of obese rats.

Objective:To investigate the effects of titanium spcimens with different surface character on the proliferation and mRNA expression of IL-6 and Cbfα1 in osteoblasts.Methods:Titanium surface was treated by smooth pretreatment(PT),sandblast and acid etch(SLA)and anodic oxidation(AD)respectively.The morphology and the elements analysis of the spcimens were inspected and detected by SEMand EDS.The surface contact angle was measured by contact angle meter.MC3T3-E1 cells were cultured on the titanium surface and cells cultured on tissue culture plate were served as the control group.The proliferation was measured by MTT assay.The mRNA expression of IL-6 and Cbfα1 was quantified by RT-qPCR.Results:The sample surface in PT group showed scrat-ches,in SLA group showed multiple three dimensional structure,in AD group exhibited porous structure.The elements of the sample surface of group PT,SLA and AD were Ti,Ti/Al and Ti/O respectively;the contact angles were 54.47°±3.33°,75.42°±8.32° and 38.91 °±4.00°respectively(P<0.05).The cells in AD group showed higher proliferation than those in PT and SLA groups(P<0.05).In AD group IL-6 mRNA expression decreased and Cbfα1 mRNA increased more than in PT and SLA groups(P<0.05). Conclusion:Titanium spcimens treated with AD may promote cell proliferation,decrease IL-6 mRNA expression and increase Cbfα1 mRNA expression in MC3T3 cells.Implats treated with AD might have some advantages in early osseointegration.

Objective To establish the rapid purification of Coxsackievirus A16 using ultracentrifugation .And To prepare and i‐dentify the neutralizing monoclonal antibody against CA16 .Methods The CA16 culture supernatant was harvested and then con‐centrated by 100K capsule .The concentration of CA16 was purified by cesium chloride ultracentrifugation .Purification of CA16 were identified by transmission electron microscopy .BALB/c mice were immunized with inactivated CA16 .Spleen cells were harves‐ted and fused with SP2/0 myeloma cells ,hybridoma cell strain secreting mAb against CA16 were objected to screening .Character‐ization of the prepared mAb were analyzed by ELISA and microneutralization assay .Results The purified CA16 method of cesium chloride gradient ultracentrifugation was established ,TEM analysis was showed that CA16 particles have icosahedral structure ,the diameters of the viral particles were approximately 20-30 nm .Two hybridoma cell strains secreting mAb against CA16 were ob‐tained ,the subtypes of two mAbs were IgG2a ,the binding titers of Anti/CA16/5 and Anti/CA16/10 were 103 and 104 respectively . Neutralizing titer of the two mAbs were 1∶256 and 1∶1 024 respectively .Conclusion Establishment method of cesium chloride gradient ultracentrifugation was performed to purify CA16 ,the two mAbs with neutralizing ability to against CA16 may become ap‐plication of treatment and vaccine .