Objective To investigate the molecular types and related clinical features of methicillin-resistant Staphylococcus aureus (MRSA) in Jingzhou area, Hubei Province.Methods A total of 80 MRSA strains confirmed by mecA gene were isolated from inpatients in Jingzhou Central Hospital of Hubei province during January and December 2014. Vitek 2 Compact was used for antibiotic susceptibility test . Staphylococcus protein A (SPA) types and Staphylococcal cassette chromosome mec (SCCmec) genotypes were detected by multiplex polymerase chain reaction ( PCR ) and gene sequencing . Panton-valentine leucocidin ( pvl) gene of the strains was detected by PCR .Chi-square test and Wilcoxon test were used for data analysis .Results There were 16 spa types in 80 MRSA isolates , in which t030 and t437 were the most prevalent ones accounting for 50.0% ( 40 strains ) and 28.8% ( 23 strains ) of the total strains, respectively.There were 77 strains of SCCmec type Ⅰ-Ⅴ, in which SCCmecⅢ and SCCmecⅣ were the most prevalent ones accounting for 45.0% (36 strains) and 35.0% (28 strains), respectively.t030 was the main spa type in isolates of SCCmecⅢ(33/36, 91.7%), while t437 was the main spa type in isolates of SCCmecⅣ(20/28, 71.4%).Patients infected with t030/SCCmecⅢMRSAs were with higher ages than those infected with t437/SCCmecⅣMRSAs (T=446.500 and 607.500, P<0.01).Patients infected with t030/SCCmecⅢ MRSAs were mainly from surgical wards and intensive care unit ( ICU ) , while those infected with t437/SCCmecⅣ MRSAs were mainly from pediatrics wards , and there were significant differences in ward distribution between two groups (χ2 =33.724 and 29.768, P <0.01).Seventy percent and above strains of t030/SCCmec type Ⅲ were resistant to rifampin, erythromycin, clindamycin, tetracycline, levofloxacin, moxifloxacin, ciprofloxacin and gentamicin .Strains of t437/SCCmec type Ⅳwere resistant to erythromycin , clindamycin and tetracycline , but were sensitive to most non-β-lactam antimicrobial drugs (with resistance rates <20%).Virulence gene pvl was found in 11 strains (13.8%), in which 7 were strains of t437-SCCmec typeⅣ.Conclusions MRSAs in Jinzhou are of various genotypes , in which t030-SCCmecⅢand t437-SCCmecⅣare the most prevalent ones .Strains of t030-SCCmec typeⅢare usually multiple-drug resistant , mainly seen in elderly patients in surgical wards and ICU .Strains of t437-SCCmecⅣare sensitive to most non-β-lactam antimicrobial drugs , and its infection is mainly seen in children and young people .

Objective To investigate the epidemiology and antibiotic resistance of communityassociated and hospital-associated meticillin-resistant Staphylococcus aureus (CA-MRSA and HA-MRSA) in Jingzhou.Methods A total of 159 MRSA isolates were successively collected from patients in Jingzhou Central Hospital during January 2012 and December 2013.The minimum inhibitory concentrations of 16 antimicrobial agents against 159 MRSA isolates were detected.SCCmec types of the strains were detected by multiplex PCR,and the homology of the strains was analyzed using pulsed field gel electrophoresis (PFGE) and cluster analysis of antibiogram.WHONET 5.6 and SPSS 19.0 were used for data analysis.Results Among 159 MRSA strains,131 were hospital-associated,and 28 were community-associated,which accounted for 82.4％ and 17.6％,respectively.There were significant differences in the age of patients,ward distribution,specimen type,length of stay,length of anti-infection treatment,type of infection and underlying diseases between patients with CA-MRSA or HA-MRSA infections (x2 =19.103,31.372,59.756,71.703,54.153,59.756 and 54.232,all P ＜ 0.01).No vancomycin,linezolid,tigecyeline and nitrofurantoin resistant strains were found,but all strains were resistant to penicillin,cefoxitin and oxacillin.HA-MRSA had higher resistance rates to moxifloxacin,levofloxacin,rifampicin,ciprofloxacin and gentamicin than CA-MRSA (x2 =30.179,27.352,28.523,28.523 and 25.987,all P ＜ 0.01),but its resistance rates to erythromycin and clindamycin were lower (x2 =13.106 and 11.743,both P ＜ 0.01).Among 159 MRSA strains,12 (7.5％) were of SCCmec type Ⅱ,113 (71.1％) were of SCCmec type Ⅲ,26 (16.4％) were of SCCmec type Ⅳ,and 8 were of undifferentiated type.The predominant SCCmec types were type Ⅳ for CA-MRSA (26/28,92.9％) and type Ⅲ for HA-MRSA (113/131,86.3％),respectively.Six PFGE patters were found in 49 HA-MRSA isolates from ICU,and the predominant patters were A1 (24,49.0％),A2 (9,18.4％) and B (9,18.4％).Cluster analysis of antibiogram showed that three groups of HA-MRSA were of high correlations,and they were of PFGE patter A1,A2 and B,respectively.Conclusions HA-MRSA is the predominant MRSA in Jingzhou area,and it is different from CA-MRSA in the age of patients,ward distribution,type of infection and antibiotic resistance.Most HA-MRSA strains are of type SCCmec Ⅲ,and may cause epidemic outbreak in ICU.

In order to investigate the chemical and morphornetrical properties of the neuronal precursor cells derived from thesubventricular zone(SVZ) of the postnatal rat forebrain in vitro. The cell-type specific antibodies were used for the immunocy-tochemical staining ,and the morphometric parameters which were the mean soma diameter and the ellipticity index (i. e. , thesmallest soma diameter divided by the largest soma diameter) of every SVZ-derived cell were measured for identifying the pheno-types of the SVZ cells in vitro. The experiment animals were SD rats (weights: 100～ 150 g), the SVZ cells derived from thepostnatal rats were cultured on poly-D-lysine-coated 24-well glass chamber slides in the Neurobasal Medium supplemented withB27 in 5% CO2 at 37 C. The following results were obtained.. At 1 day in vitro, almost all SVZ cells (〉90%) from the postna-tal rat forebrain expressed Tujl, an antibody that recognizes neuron-specific tubulin. Likewise, the preponderance of the SVZcells expressed the polysialylated neural cell adhesion molecule (PSA-N-CAM) ; The majority of the SVZ Tujl-positive cells cul-tured were the cells that had oval-shaped bodies with two short, unbranched processes protruded from every two poles, theirmean soma diameter were 8.42±1.03μm and their ellipticity index were 0.57±0.12. Meanwhile, there were approximately20% of the SVZ cells in culture that were sphere-shaped cells with mean soma diameter 7.20±l.04 μm , and it might be observed that these cells connected with one another. As the time in culture went on, these sphere-shaped SVZ-derived cells alsotransformed to oval-shaped ones as described above, but it could be observed that the cells were still connected in the processesof them. By 3 and 5 days in culture, the SVZ cells had larger cell somas (average diameter 9. 07±1.07 μm), and often consider-ably longer processes but still with few branches. Immunocytochemical staining revealed that the majority of the SVZ cells in cul-ture remained Tujl-positive, PSA-N-CAM-positive. By 7 days in culture, the Tujl-positive cells in culture showed remarkablemorphological changes, and possessed typical neuronal phenotypes, which had more larger cell somas (average diameter 12.8 ±1.13 μm), and had more longer, branched processes. Our results showed that the SVZ in the postnatal SD rats contained theneuronal precursor cells which were PSA-N-CAM-positive and could differentiate into new neurons in vitro.

Objective To observe the effect of curcumin combined with cisplatin on the proliferation of human tongue squamous cell carcinoma Tca?8113 cells,and explore the possible mechanism of their anti?tumor effect in vitro. Methods In this study,the samples were divided into four groups:curcumin,cisplatin,curcumin combined with cisplatin,and control. Detection of rate of inhibition of Tca?8113 cell proliferation was carried out by MTT assay. This was followed by observation of the morphological changes of the nuclei by Hoechst 33258 fluorescence staining. Subse?quently,cellular apoptosis was assayed by flow cytometry,and expression of Notch1 and epidermal growth factor receptor(EGFR)was examined by Western blotting. Results Results of MTT assay showed that curcumin combined with cisplatin inhibited the proliferation of tongue squamous cell carcinoma to a greater extent than either curcumin or cisplatin alone(P<0.05). Flow cytometry analysis indicated that,unlike curcumin or cisplatin alone,curcumin combined with cisplatin noticeably induced apoptosis(P<0.05). Results of Western blotting revealed that the expres?sion of Notch1 was upregulated,whereas the expression of EGFR was downregulated to a greater extent in the control group than in cells treated with curcumin or cisplatin alone. Unlike curcumin and cisplatin groups,the combined group revealed statistically significant expressions of Notch1 and EGFR(P<0.05). Conclusion Curcumin and cisplatin have a combined effect on inhibition of proliferation of human tongue squamous cell carcinoma Tca?8113 cells. The mechanism may be related to upregulation of the Notch1 signaling pathway and downregulation of the EGFR signal?ing pathway.

Objective To explore the influence of peritumoral edema (PTE) on the tendency of recurrent location and morphological character after total resection using MRI. Methods MRI data was collected from 43 patients with recur-rent brain glioma after total resection from four clinical centers and then the influence of of PTE on recurrence patterns af-ter total resection was retrospectively analyzed based on the T2 weighted image. Results The PTE had a significant influ-ence on the recurrent patterns of brain gliomas after total resection. When PTE was mild, the shapes of recurrent gliomas tended to be focal (6/8) and the recurrent locations tended to be local (5/8). When PTE was severe, the shapes of the recur- rent gliomas tended to be spread(30/35 and the recurrent locations tended to be distant (25/35), followed by marginal (7/35), In addition, the morphological patterns and locations of recurrent gliomas were significantly different among different PTE types (all P<0.001). When PTE was ring shape, the shapes of recurrent gliomas tended to be focal (7/9) and the recur-rent locations tended to be local (6/9), followed by marginal (2/9) and distant (1/9). When PTE was irregular shape, most of recurrent locations tended to be distant (25/34), followed by marginal (7/34) but rarely local (2/34). Conclusions The de-grees and the types of brain glioma PTE can significantly influence the locations and morphological patterns of recurrent gliomas after total resection.

Objective To explore the relationship between serum homocysteine (Hcy) level before coronary angiography(CAG) and contrast induced nephropathy (CIN) after CAG.Methods We included 2264 cases of suspected coronary heart disease from May 2013 to May 2016 and all patients received CAG examination.According to whether CIN has developed or not after CAG, the patients were divided into the non-CIN group (n=2162) and the CIN group (n=102).We analyzed and compared the clinical baseline data, serum Hcy and creatinine (Cr) levels and the estimated glomerular filtration rate between the 2 groups eGFR.Results Patients in the non-CIN group were younger and with less comorbidities of diabetes and chronic kidney disease (all P<0.05).The volume of contrast media consumed in the non-CIN group was less than the CIN group [(122±21)ml vs.(147±24)ml, P=0.012).Hcy level in the non-CIN group (12.81±6.71) μmol/L was lower than that in the CIN group (21.74±11.9)μmol/L before CAG (P<0.05).No significant differences in serum Cr level and eGFR before CAG (P>0.05).At 72 hours after CAG, Cr level of the non-CIN group (69.34±19.54 μmol/L) was lower than that of the CIN group (87.34±21.38) μmol/L (P<0.05).eGFR was higher in the non-CIN group (79.34±19.54)ml/min than that in the CIN group (67.34±21.38)ml/min (P<0.05).Linear regression analysis showed that Hcy level before CAG were positively correlated with Cr level after CAG (r=0.547,P<0.01) and negatively correlated with eGFR after CAG (r=-0.271,P<0.01).Conclusions Hcy level before CAG can be used as one of an effective parameter to predict CIN.

OBJECTIVE To establish a method of loop-mediated isothermal amplification(LAMP) for detecting Staphylococcus aureus in positive blood culture bottles.METHODS Genomic DNA in 293 positive blood culture bottles was extracted by guanidine hydrochloride and benzenemethanol,then genes ssa and mecA were amplified by LAMP to identify S.aureus.Finally,the results of LAMP were compared with the results of traditional method.RESULTS Twenty-two strains of S.aureus were detected in 293 positive blood culture bottles by LAMP method.Compared with traditional method,the sensitivity and specificity of LAMP method were both 100%,respectively,the detection could be finished in an hour.CONCLUSIONS The LAMP-based assay is simple,rapid,sensitive and specific which can be used to detect S.aureus in positive blood culture bottles rapidly.

OBJECTIVETo establish a fluorogenic probe quantitative RT-PCR (FQ-RT-PCR) method for detection of bcr/abl mRNA fusion gene expression level in leukemia cells, and provide a useful tool for leukaemia diagnosis and minimal residual disease inspectation.

METHODThe conventional RT-PCR was used to amplify bcr/abl gene from cultured K562 cells, the quantitative standard template was constructed with A-T clone method. The fluo-rogenic quantitative RT-PCR method by using Applied Biosystems 7700 Sequence Detector for detecting the expression of bcr/abl fusion gene was successfully. The sensitivity, stability and repetitiveness of this method was determined. The peripheral blood samples from 14 CML patients, one of whom before and one month after bone marrow transplantation (BMT) and 4 cases of ALL in the early stages were detected.

RESULTSThe sensitivity of FQ-RT-PCR for detecting bcr/abl fusion gene was about 10(-5) micro g RNA from K562 cell and 10 copies recombined plasmid. The repetition CT value (cycle threshold) and the coefficient variation (CV) among tubes and batches were 2.0% and 3.7%, respectively. The median bcr/abl fusion gene expression level of 14 CML patients was 5.15 x 10(4) copies/ micro g RNA. The products analyzed by electrophoresis showed that 11 cases were b2a2 and 3 cases b3a2. 1.2 x 10(5) copies/ micro g RNA in one CML patient before BMT was changed to 2.3 x 10(3) copies/ micro g RNA one month after BMT. B2a2 was observed in one of the four (25.0%) patients with ALL, and its level of expression was 8.2 x 10(5) copies/ micro g RNA.

CONCLUSIONThe established FQ-RT-PCR method is sensitive, specific, reliable, accurate and good at repetitiveness. The results expressed in copies were easy for evaluation and comparation. Two different bcr/abl fusion gene form - b2a2, b3a2 can be detected by the method. It can be widely applied to diagnosis and detection of minimal residual disease for CML and some ALL patients.

Objective Exploring clinical prediction value of the ischemia modified albumin (IMA) for acute coronary syndrome (ACS) patients after the percutaneous coronary intervention (PCI).Methods A total of 109 cases of the PCI postoperative patients of ACS at the Panggang General Hospital from January 2010 to July 2012 were included.IMA was determined within 6 hours after admission,and standardized treatment after PCI.After a 6-month follow-up,they were divided into the event group and non-event group according to cardiovascular events occurrence.Results 101 cases were followed-up,including 26 cases of cardiovascular events (25.74％) and 75 cases of no cardiovascular events (74.26％).The left ventricular ejection fraction of the event group [(45 ± 7)％] was significantly lower than the non-event group [(52 ± 10) ％] (t =1.894,P ＜ 0.05).Serum IMA of the event group[(105.51 ± 13.26) U/ml]was significantly higher than the non-event group [(85.18 ± 11.36) U/ml] (t =7.3518,P ＜0.01).After controlling other cardiovascular risk factors,IMA was still independent risk factors for cardiovascular events (OR =1.69,95％ CI: 1.18 ～ 2.13,P =0.01).Conclusions IMA have very good clinical prediction value of cardiovascular events occurrence for ACS after PCI.

Objective: To analyze Internet behaviors of children and adolescents and their influencing factors, and to provide scientific basis for health education guidance and prevention measures. Methods: A stratified cluster sampling method was used. From January to June 2019, 30 955 primary school students in 16 provinces (municipalities), from eastern, central, and western economic zones of China were selected to conduct a questionnaire survey, regarding general demographics, diet and exercise, tobacco, alcohol and sleep, Internet and electronics usage, etc. Results: Among them, 43.1% of students had never been online with girls (44.7%) higher than boys (41.8%)(χ2=21.04, P<0.01); 1.3% of students reported online time for more than 4 hours a day, boys were significantly higher than girls(χ2=15.87, P<0.01); Internet addiction was detected in 2.4% of elementary school students, with boys (3.0%) higher than girls(1.7%)(χ2=331.77, P<0.01), and the trend increases with grade. With sixth grade (3.3%) were significantly higher than children from the first grade (0.8%) and other grades(χ2=163.96, P<0.01). Children’s tendency to Internet addiction was 11.9%, of which boys were 13.0% and girls were 10.6%, boys were significantly higher than girls(χ2=117.66, P<0.01). Pearson correlation analysis revealed that, after controlling gender age, boarding school, family relationship, single parents and regrouped families, smoking, drinking sugary drinks, eating fried foods, and left-behind were positively related to Intenet addiction(r=0.06, 0.07, 0.27, 0.44, 0.07, 0.11, 0.09, P<0.01), while medium and high intensity sports, effective physical education, health education in school, AIDS education and parental restrictions on TV and computer games, were negatively associated with internet addiction(r=-0.74, -0.65, -0.04, -0.05, -0.63, P<0.01). Conclusion The internet addiction of Chinese primary school students is common and increases by year. High-intensity, high-density sports, effective physical education, and health education could help prevent Internet addiction among children.