Objective To explore the prevalence of qnr gene in gram-negative bacteria isolated from Renmin Hospital of Wuhan University. Methods qnr gene was detected by PCR in non-duplicate E.coli (n=129), E.cloacae (n=10) and K.pneumoniae (n=29) isolates.PCR technique was used to amplify the gene encoding type I integrons.Kirby-Bauer disk diffusion method was used to test the in vitro suscepti- bility of the isolates to 16 antimicrobial agents.ESBLs were investigated according to phenotypie screening and confirmatory test recom- mended by NCCLS.MIC values of ciprofloxacin were determined for qnr gene positive strains.Results qnr gene were identified in 6 clinical isolates including 5 strains of E.coli and I E.cloacae, qnr gene was not found in K.pneumoniae isolates.The 6 qnr-positive clinical iso- lates were positive for type I integrons.All the 6 strains were susceptible to imipenem but resistant to many other drugs tested.Only 2 of the E.coli isolates were susceptible to ciprofloxacin.Conclusions In Hubei Province, the prevalence of qnr is confirmed.The serious situa- tion of multidrug-resistance in clinical isolates with qnr gene requires careful monitoring of qnr-carrying isolates.

Adenoma, Villous ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Lymphoma ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Myosins ; metabolism ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Rhabdomyosarcoma, Alveolar ; metabolism ; pathology ; surgery ; Ureteral Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Objective To evaluate the impact of ghrelin on depleted uranium ( DU)-induced damage of the osteoblast MC3T3-E1. Methods MC3T3-E1 cells were treated with different doses of ghrelin for 1 h before DU (500 μM) treatment. After 24 hours,the cell via-bility,intracellular tartrate-resistant acid phosphatase (StrACP),alkaline phosphatase (AKP),osteoprotegerin (OPG),solvable receptor acti-vator of nuclear factor-κB ligand ( sRANKL) ,catalase ( CAT) and reactive oxygen species ( ROS) were measured. Results After DU expo-sure,ghrelin pretreatment increased the cell viability and CAT levels,and reduced intracellular StrACP,AKP,sRANKL/OPG and ROS in a dose-dependent manner. Conclusion Through maintaining the balance of OPG/RANKL and reducing the oxidative stress,ghrelin could pro-tect against DU-induced damage of MC3T3-E1 cells.

Objective To establish the HeLa cell line that can stably express EYFP fluorescent protein as the model for anion channel blocker (halide ion) screening ,which lays the foundation for high throughput screening of anion channel blocker (halide ion) .Methods Through gene recombination technology ,a new lentivirus vector which can express mutant protein YFP (EYFP‐H148Q/I152L) and puromycin resistance ,was built .The mixture of lentivirus vector and packaging plasmid was transfected into 293T cells to produce lentivirus particles . After infection of HeLa cells by the lentivirus particles ,puromycin was used to screen the cells as YFP‐positive HeLa cell line .Then cell amplification was carried out after purification and efficiency of EYFP‐H148Q/I152L was further detected by Real‐time quantitative PCR (RT‐PCR) and Western blot .We then verified the activity of EYFP‐HeLa transfected cell line as a screening model of anion channel blocker .Results Gene sequencing verified that EYFP‐H148Q/I152L was successfully inserted into lentivirus vectors .RT‐PCR and Western blot results showed that the target gene was overexpressed in HeLa cells . The specific yellow fluorescence of EYFP of HeLa cells could be observed under fluorescence microscope with the efficiency of nearly 100% . I- (low permeability ) solution stimulated the opening of anion (halogen) channels ,and the yellow fluorescence was quenched by I - flow into cells . Conclusion The EYFP‐HeLa cell line can stably express EYFP yellow fluorescent protein and is sensitive to the internal flow of I - .Therefore ,it can be used as an ideal screening model of anion channel blocker (halide ion) .

Objective To construct an eukaryotic expression vector with human adipose most abundant gene transcript 1 (APM1) gene,and to investigate the transfection and expression of pCDEF-APM1 eukaryotic expression plasmid in HEK293 cells.Methods pCDEF-APM1 eukaryotic expression plasmid was constructed by DNA recombinant method.Expression vector pCDEF-APM1 was transfected into HEK293 cells with Effectene reagent.The level of human adiponectin protein in the supernatant of cell culture media was detected with double antibody sandwich ELISA.Results The sequence of DNA fragment from constructed pCDEF-APM1 plasmid was identical to that published in GenBank.There was raised human adiponectin protein level in culture supernatant of HEK293 cells tnmsfected with pCDEF-APM1.Conclusion The pCDEF-APM1,an eukaryotic expression plasmid for APM1 gene is successfully constructed.High protein expression of adiponectin can be obtained in HEK293 cells transfected with pCDEF-APM1 eukaryotic expression plasmid.

OBJECTIVETo observe changes of gonad functions of Gan-qi stagnation (GS), Pi deficiency (PD), Gan-qi stagnation Pi deficiency (GSPD) model rats, and the effect of Chaishu Sijun Decoction (CSD) on them.

METHODSRats were randomly divided into 7 groups according to romdom digit table, i.e., the normal control group, the GS model group, the GS medication group, the PD model group, the PD medication group, the GSPD model group,and the GSPD medication group, 10 in each group. Rats in the GS model group, the PD model group, and the GSPD model group were treated with chronic restraint, improper diet +excessive fatigue, chronic restraint +improper diet +excessive fatigue. The model was established for 4 successive weeks. Starting from the 15th day of modeling, CSD at the daily dose of 3.57 g/kg was given by gastrogavage to them for 14 successive days. Equal volume of distilled water was given by gastrogavage to rats in each model group and the normal control group for 14 successive days. The blood contents of gonadotrophin releasing hormone (GnRH), follicular stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and testosterone (T) were detected in rats of each group.

RESULTSCompared with the normal control group, there was statistical difference in GnRH, T, E,, and FSH in the GS, PD, and GSPD model groups (P < 0.05, P < 0.01). The content of LH was elevated in the GS model group (P < 0.05) and declined in the GSPD model group (P < 0.01). Compared with the GS model group, the contents of FSH, LH, and T decreased and E2 increased in the PD model group (all P < 0.05); the contents of FSH and LH also declined in the GSPD model group (P < 0.05). Compared with the PD model group, the T content increased and FSH decreased in the GSPD model group (all P < 0.05). Compared with each corresponding model group, the FSH content decreased (P < 0.01) and LH increased in the GS medication group; the T content increased, E2 and LH decreased (P < 0.05, P < 0.01) in the PD medication group; the T content decreased (P < 0.01), GnRH, E2, FSH, and LH increased (P < 0. 05, P < 0.01) in the GSPD medication group.

CONCLUSIONSThere exist different degrees of abnormal function of the gonad axis in the GS, PD, and GSPD models. CSD had certain regulatory effect on the 3 syndromes. Of them, it showed a more comprehensive role in improving the gonad function axis. Results of this experiment had provided the experimental evidence for higher correlation between CSD and GSPD syndrome.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Gonadal Steroid Hormones ; blood ; Gonads ; drug effects ; Male ; Rats ; Rats, Wistar

Objective:To analyze the mean levels of skeletal muscle mass and strength in elderly patients with type 2 diabetes mellitus(T2DM), and to investigate the effects of chronic inflammatory factors and oxidative stress on them.Methods:A cross-sectional study was conducted on 120 patients with T2DM aged over 60 years and 126 elderly patients without diabetes(the control group). Skeletal muscle mass, strength and serum levels of chronic inflammatory factors interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and 8-hydroxy-2′-deoxyguanosine(8-OHdG)were determined, and their effects on skeletal muscle mass and strength in elderly patients with T2DM were analyzed.Results:Compared with the control group, grip strength decreased in elderly patients with T2DM(25.03±7.85)kg vs.(29.52±7.73)kg( P<0.01), and skeletal muscle mass decreased(21.36±5.46)kg vs.(22.01±5.22)kg with no significant difference( P>0.05). Serum levels of 8-OHdG were higher in elderly patients with T2DM than in the control group(3.08±0.26)ng/L vs.(2.59±0.16)ng/L( P<0.01). Correlation and regression analysis results showed that 8-OHdG was an influencing factor for muscle strength in elderly patients with T2DM( R2=0.457)and that height and weight could be influencing factors for skeletal muscle mass in elderly patients with T2DM( R2=0.822). Conclusions:Skeletal muscle mass and strength decline in elderly T2DM patients, probably as a result of increased levels of oxidative stress.These findings may serve as evidence for sarcopenia intervention in elderly T2DM patients.

Objective To explore the mastery of relevant knowledge and skills for autism rehabilitation teachers in Beijing private rehabilitation agencies. Methods 51 autism rehabilitation teachers were selected from 10 Beijing private rehabilitation agencies and tested with self-prepared questionnaire. Results Rehabilitation teachers believed that professional knowledge, rehabilitation skills and special education basics ranked the former three positions as for knowledge importance; teachers' basic skills, professional knowledge and special education basics ranked the former three positions as for knowledge mastery. There were difference in academic background and business life. 95% teachers believed that the training form was knowledge plus case plus guidance. The best training time was weekends or summer and winter vacations. The curriculum expected mainly focused on behavior modification, class management, and training system that generally used for children with autism. Conclusion Autism rehabilitation teachers in Beijing private rehabilitation agencies urgently need training in the aspects of expertise, rehabilitation skills and so on.

Objective To evaluate the predictive value of ABCD2 scoring system in cerebral infarction and death events in the medium-term after transient ischemic attack (TIA) of Chinese population. Methods One hundred and seventy-nine patients with TIA having complete clinical data,admitted to our hospital from January 2008, were chosen in our perspective study. The ABCD2 scale was applied to all the patients and used to observe the 18-month cerebral infarction risk events; according to these data, patients after TIA were divided into cerebral infarction group and non-cerebral infarction group; To the end of 18 months, according to the death event, the patients were divided into survival group and death group. The age, level of blood pressure, clinical features, duration of symptoms and history of diabetes were collected and compared between each 2 groups; the cutofflevel of ABCI2 scale for anticipating cerebral infarction risk and mortality was evaluated according to receiver operating characteristic (ROC) curve. Results Among the studied 179 patients, 52 patients appeared cerebral infarctions within 18 months of TIA; patients in the cerebral infarction group had significantly higher percentage of patients with blood pressure ≥ 140/90 mm Hg (86.5％), unilateral weakness (42.3％),duration of symptoms ≥60 minutes (55.8％), or duration of symptoms 10 to 59 minutes (40.4％), and diabetes (80.8％) as compared with patients in the non-cerebral infarction group (P＜0.05). According to the ROC curve, area under curve was 0.874 of ABCD2 scale for anticipating cerebral infarction risk, 95％confidence interval (CI) was 0.817-0.931 and the cutoff level of ABCD2 scale for anticipating medium-term cerebral infarction risk was 4.5. Among the studied 179 patients, 35 patients died within 18months of TIA; no significant difference in the scores of ABCD2 scale was noted between survival group and death group (P＞0.05); according to ROC curve, area under curve was 0.492 of ABCD2 scale for anticipating mortality, and 95％ CI was 0.389-0.596. Conclusion ABCD2 scale has predictive value in cerebral infarction event, while not in death event in the medium-term of TIA.

Objective To investigate the role of HBO on the change of levels of ACA and thrombomodulin among parents with ischemic encephalopathy. Methods Patients were divided into three groups: group A (patients with atherosclerosis and thrombosis),group B (patients with cardiogenic embolism) and group C (patients with small artery lesion). All patients were treated with HBO and regular drugs. And then sss scores were recorded and levels of ACA-IgG and thrombomodulin were detected by ELISA before and after therapy of HBO. Results The index of ACA-IgG and lelvel of TM in A,B and C group were higher than those in control group,there are differences among them(P＜0.05),and the level of them were higher in B group than in A and C group.There were significant differences in SSS score for brain function using different therapies in A、B and C group (P＜0.05), but there are no differences among them(P＞0.05).There were differences in the index of ACA-IgG and thrombomodulin using different therapies in A,B and C group(P＜0.05),however,there were no differences among them (P＞0.05). Conclusion HBO can alleviate clinical symptoms and improve recovery of neuron by decreasing the level of ACA-IgG and thrombomodulin in blood,but the effect is not significantly different for ischemic encephalopathy caused by different factors.