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 .

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 .

Objective To compare the efficacy of supraclavicular approach and infraclavicular approach in ultrasound-guided brachial plexus block(BPB).Methods One hundred and twenty patients,ASA Ⅰ-Ⅲ,aged 18-80 yr,scheduled for upper extremity surgeries of both gender,were randomized into two groups(n =60):supraclavicular group(group SCB)and infraclavicular group(group ICB).The anesthetic mixture consisted of ropivacaine 0.375％ and lidocaine 1％ in equal volumes with epinephrine 1∶200 000,the total dose was 0.5 ml/kg.The block performance time,duration of anesthesia and success of anesthesia(surgery was accomplished without supplementary block)were recorded.A blinded observer assessed pinprick sensory block in the seven distal nerve territories(axillary,radial,musculocutaneous,median,ulnar,medial antebrachial and medial brachial cutaneous nerves)every 5 min up to 30 min after injection.Success of nerve block,side effects and complications were recorded during and after operalion.Results Group ICB was superior in success rate of anesthesia and nerve block of ulnar,medial antebrachial and medial brachial cutaneous nerves,the rate of parasthesia was lower and the block performance time was longer compared with group SCB(P ＜ 0.05).No major complications occurred in both groups.Conclusion Under ultrasound guidance,infraclavicular BPB is superior to supraclavicular approach.

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.

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: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

Objective To evaluate the changes of respiratory mechanics and central drive in sleeping patients with chronic obstructive pulmonary disease (COPD).Methods A total of 14 patients with moderate to severe COPD (age 63.7± 6.4 years) and 10 healthy volunteers,admitted to our hospital from October 2010 to October 2011,were chosen in our study.After performing routine pulmonary function test,all subjects were monitored by respiratory mechanics,diaphragm electromyogram (EMGdi) and polysomnography (PSG) in waking and sleeping state.The different physiological parameters were recorded and calculated on a continuous basis in time sequence for 30 minutes.Results In sleeping state as compared with waking state,there were no differences(P＞0.05) in respiratory rate(RR) and the ratio of inspiratory time to duration of one breath (Ti/Ttot),but there were reductions (P ＜ 0.05) in the tidal volume (VT),minute ventilation (VE),mean inspiratory volume per second(VT/Ti),dynamic lung compliance(CLdyn),root mean square (RMS)and pulse oxygen saturation(SpO2).Meanwhile there were significant increases (P＜0.05) in airway resistance(Raw) and pressure-time product(PTP) in COPD group.In normal group,there were no differences (P＞0.05) in all above physiological parameters in sleeping state compared with waking state.Conclusions There are abnormal changes in respiratory mechanics characterized by increased airway resistance and work of breathing,meanwhile in reduced central drive and requirement of ventilation,which may be the main reasons for hypopnea and hypoxemia in sleeping COPD patients.