Objective To detect ESBLs in E.coli and K.pneumoniae with appropriate method.Methods The suspiciously produced ESBLs were detected by standard screen test among 110 E.coli and 84 K.pneumoniae. ESBLs possessing strains were confirmed by E test, single disc diffusion test with clavulanic acid or sulbatan, and two kinds of double disc synergy test. Results There was a significant difference between the detected rate of ESBLs in E.coli (16/26) and K. pneumoniae (4/20) by E test( P 0.05). The single disc diffusion test with clavulanic acid or sulbatan was applicable to confirm ESBLs in K. pneumoniae (12/20). Conclusions Single disc diffusion test is a sensitive, convenient and inexpensive method to confirm ESBLs in E.coli and K.pneumoniae in clinical laboratory.

Objects To observe the TEM gene characteristic of Acinetinbacter lwoffi strains which were resistant to most kind of antibiotics and to compare their sequences with that of Escherichia coli Methods Two strains of A lwoffi, JN18 and JN70,were isolated from sputum of patients′ who suffered from respiratory infection TEM, SHV, OXA, IMP and CTX M genes were tested by PCR TEM sequences of JN18 and JN70 were detected by ABI automated sequencer and were analysed by DNAStar software to compare the differences with E coli TEM genes that had been published in GenBank Results Detected sequences of A lwoffi JN18 and JN70 strains were 1012 bp and 887 bp, respectively They coded regions of 832 bp and 772 bp for TEM of the two strains that were 98 2% identical In other hand, there were 14 pair bases differently in TEM regions The TEM sequence of JN18 was 98 72% identical to that of E coli TEM on the average, which was over 99% to TEM1D, TEM 70, TEM 76, TEM 77 and TEM 95 of E coli The divergence of TEM genes was 1 26 between A lwoffi JN18 strains and E coli JN70 strain had higher identity (98 93%), which reached 99 5% to TEM1D, TEM1F and TEM84 of E coli As JN18 strain, TEM gene of JN70 had very small divergence (1 06 ) with E coli Conclusion JN18 and JN70 strains of A lwoffi isolated from patients′respiratory tract in Jinan shared most of sequences with E coli TEM76 and 84 However, there were many mutant sites in them

Objective To evaluate curative effect and safety of moxifloxacin in treatment of acute pelvic inflammatory with Meta analysis.Methods The database of Pubmed, MEDLINE, Cochrane, China Biology Medicine disc (DBMdisc), China National Knowledge Infrastructure (CNKI), Wanfang med online and VIP were retrieved, and randomized clinical trials of moxifloxacin in treatment of acute pelvic inflammatory which were open literatures published at home and abroad were collected from January 2000 to December 2014.The literatures which were accorded with the criterion were chosen and analyzed by Review Manager 5.1.7.Results The results showed that there were 14 literatures which were homogenous corresponded with the criterion, changeless benefit model statistics and analysis showed that the odds ratio (OR) was 3.76(95%CI 2.14-6.59, P<0.01).And there were 5 studies on bacterial removal showed that the OR was 2.34(95%CI 1.46-3.75, P<0.01).There were 12 studies on incidence of adverse reactions showed that the OR was 0.5(95%CI 0.30-0.86,P=0.01).Conclusion Meta analysis displayed that moxifloxacin is effective for acute pelvic inflammatory.But the high quality RCTs were too few in domestic studies in treatment of the acute pelvic inflammation with moxifloxacin and the overall level of studied quality needs improvement urgently.

Aim To establish a mouse model of AIDS and observe the effects of Combivir,an anti-AIDS drug.Methods The model was set up by infecting infant mice with L6565 murine leukemia virus.Quantitative RT-PCR method was used to measure viral load in plasma.The percentage of CD4~+ and CD8~+ lymphacytes was estimated by flowcytometric method.The number of WBC,RBC and marrow karyote was counted.indexes of immune organs were weighed and calculated.Results Compared with control,the viral load in plasma in the model was at high level.The percentage of CD4~+ lymphocyte,the ratio of CD4~+/CD8~+,the number of RBC and marrow karyote,and thymus index were significantly decreased in the model,while the number of WBC and spleen index was remarkably increased.Conclusion AIDS patient symptom is shown in mouse infected with L6565 murine leukemia virus.This suggests that this model is likely to be used as a mice model to evaluate the effect of anti-AIDS drugs.

AIM: To extract the effective composition from Radix Arnebiae seu Lithospermi and make it into the vagina effervescent tablet. It would be convenient and effective to treat the disease for the patients. METHODS : To sift the technology of extracting the effective composition by experiment and find out the prescription of the vagina effervescent tablet,its time of disintegration and the quantity of producing foam can accord with the principle of the vagina effervescent tablet in Chinese pharmacopoeia. RESULTS : The technology of extracting the effective composition and the prescription of the vagina effervescent tablet by experiment are practical. CONCLUSION : It is convenience for patients to take vagina effervescent tablet of Radix Arnebiae to treat their diseases

BACKGROUND: MyoD gene is one of family members of muscle transcription factors. Transfection MyoD gene can switch on the procedure of differentiation of muscles, and transit non-muscle cells into muscle cells.The MyoD gene only expresses in skeletal muscles. Based on the same contractive structure in myocardial cells and skeletal muscle cells, it is imagined that the conversion from exogenous MyoD gene-induced fibroblast in local myocardium into skeletal muscle cells that had contractive function may become another method in the treatment of congestive heart failure on clinic.OBJECTIVE: To construct and identify a recombinant adenoviral vector carrying MyoD gene for further studies on the recovery function of MyoD gene in myocardial injury.DESIGN: Single sample experiment.SETTING: Department of Cardiology, First Affiliated Hospital, Nanjing Medical University.MATERIALS: The experiment was conducted at the Laboratory of Microbiology and Immunology, Nanjing Medical University between September 2004 and September 2005. MyoD gene and non-replicating form expressive vector of adenovirus were taken as research materials.METHODS: MyoD cDNA fragments were extracted from plasmids pEMSV-MyoD with polymerase chain reaction (PCR), and PCR was used to clone the whole-length gene of MyoD. After adding CACC sequence at 5' end, MyoD gene was cloned by orient topology into transfer ventor, pENTR/D-TOPO. Objective gene was transferred into adenoviral expression vector DNA via pENTR/D-TOPO vector. The recombinant adenoviral vectors transfected into HEK293A cells by using lipofectamine were packaged and amplified.MAIN OUTCOME MEASURES: Evaluation of PCR and DNA sequencing were used for confirming the size of segment and correctness of rank of MyoD cDNA and detecting the titre of virus.RESULTS: MyoD recombinant adenovirus contained target segment with precise length confirmed by PCR and DNA sequence that was correct. The titre of virus was 1.3×1011 pfu/mL.CONCLUSION: The recombinant adenoviral vector carrying MyoD gene is constructed successfully.

Self-biting is a chronic disease, which cause wound to take effect on mink growth and pelt quality. In this study, we firstly adopted RAPD (random amplification polymorphism DNA) technique based on the reproducible 26 polymorphism primers screened from 100 random primers to analyze hereditary constitution of the samples from healthy minks and self-biting minks, respectively, at molecular level to aim to discuss the causes of self-biting. The results showed that 29 straps showed polymorphism among amplified 105 straps, of which the polymorphism rate is 27.62%. Between healthy and sick mink groups, the amplified DNA fragment through different primers indicated different distribution frequency. The similarity coefficient of mink groups is 0.8471 and genetic distance (variation) index is 0.1529. Through primer S356 (whose sequence is CTGCTTAGGG), we amplified different straps between healthy and sick mink. The amplified 1000 bp DNA fragment in the sick mink groups can preliminarily serve as molecular genetic label to distinguish from healthy and sick mink groups to gradually remove the mink individual of self-biting, achieve to purify mink groups and reduce economy loss of mink breeding industry. This work provide theoretical basis for further study on molecular breeding and disease prevention of mink.
Animals ; Base Sequence ; Genetic Diseases, Inborn ; etiology ; genetics ; veterinary ; Mink ; genetics ; Molecular Sequence Data ; Random Amplified Polymorphic DNA Technique

Objective To identify the expression of PTPRD in esophageal squamous cell carcinoma (ESCC)and analyze its correlation with pathological features and patient survival.Methods Immunohistochemistry analysis was applied to detect the expression level and location of PTPRD in 236 patients with ESCC.Clinical and pathological features were collected and a 5 years’follow-up after surgery were performed.Results Statistic analysis showed that expression of PTPRD in ESCC was lower than in normal esophageal epithelial cells (22.0% vs 57.2%,P =0.000).The expression of PTPRD was correla-ted to the differentiation grade,depth of tumor invasion and lymph nodes metastasis.The expression of PTPRD was higher in group with well differentiation,less invasion depth and no lymph node metastasis (P =0.013,0.025,0.019).The expres-sion of PTPRD was not correlated to age or gender (P =0.170,0.787).The survival analysis showed that thegroup with more PTPRD expression had better prognosis.Conclusion PTPRD was correlated to progression and prognosis of ESCC.It may be a new potential tumor suppressor gene of ESCC,and its expression level might may a useful marker for predicting prognosis for ESCC patients.

Objective To ascertain the specific activities of nigericin on inhibiting human epithelial ovarian cancer(EOC)cells,and to investigate the possible molecular mechanism of nigericin on cell migration and invasion.Methods Cell viability under different treatments of nigericin(0.312 5,0.625,1.25,2.5,5,10,20,40,80 μmol/L)on EOC A2780 and SKOV3 cell lines was examined by CCK-8 assay,with DMSO as a control.The human epithelial ovarian cancer cell lines A2780 and SKOV3 were treated with 5,10 or 20 μmol/L nigericin or with DMSO as a control.Transwell chambers was used to observe the impact of nigericin on migration and invasion of EOC cells.Western blot was used to detect the expressions of epithelial cell marker(E-cadherin),mesenchymal cell marker(Vimentin)and the epithelial-mesenchymal transition(EMT)-related transcription factors Slug,Snail,and Twist,as well as the expressions of proteins related to Wnt/β-catenin signaling pathway such as Gsk-3β,p-Gsk-3β and β-catenin under different concentration treatment of nigericin.Results CCK-8 assay showed that nigericin exhibited strong cytotoxicity on A2780 [ IC 50(16.19 ± 0.26)μmol/L,95％CI 1.077-1.341)] and SKOV3 [ IC50(11.87 ±0.21)μmol/L,95％CI 1.003-1.146] cell lines.Transwell chamber assay revealed that nigericin at different concentration(5,10,20 μmol/L)induced a remarkable reduction in the number of cells migrating through the membrane relative to the vehicle-treated controls in A2780 and SKOV3 cells [(121±9),(92±7),(59±5)/HP and(120.4±2.6),(91.8±5.5),(80.0±4.0)/HP,all P <0.05]; the invasive ability of A2780 and SKOV3 cells also inhibited [(61.2±3.7),(43.2±4.3),(23.6±2.1)/HP and(85.2±7.0),(65.2±4.6),(45.6±4.4)/HP,all P< 0.05].Western blot revealed that the increased expression of E-cadherin and decreased expression of Vimentin,Slug,Snail,Twist,p-Gsk-3β,β-catenin in EOC cells with the nigericin treatment at different concentration(5,10 and 20 μmol/L)showed concentration dependence(P < 0.05).Conclusion Nigericin may induce EMT by activating Wnt-β-catenin signaling pathway to promote the migration and invasion of EOC cells.

PURPOSE: Chronic cough in allergic rhinitis (AR) patients is common with multiple etiologies including cough variant asthma (CVA), non-asthmatic eosinophilic bronchitis (NAEB), gastroesophageal reflux-related cough (GERC), and upper airway cough syndrome (UACS). Practical indicators that distinguish these categories are lacking. We aimed to explore the diagnostic value of the fraction of exhaled nitric oxide (FeNO) and forced expiratory flow at 25% and 75% of pulmonary volume (FEF(25–75)) in specifically identifying CVA and NAEB in these patients. METHODS: Consecutive AR patients with chronic cough were screened and underwent induced sputum, FeNO, nasal nitric oxide, spirometry, and methacholine bronchial provocation testing. All patients also completed gastroesophageal reflux disease questionnaires. RESULTS: Among 1,680 AR patients, 324 (19.3%) were identified with chronic cough, of whom 316 (97.5%) underwent etiology analyses. Overall, 87 (27.5%) patients had chronic cough caused by NAEB, 78 (24.7%) by CVA, 16 (5.1%) by GERC, and 81 (25.6%) by UACS. Patients with either NAEB or CVA (n = 165, in total) were further assigned to a common group designated as CVA/NAEB, because they both responded to corticosteroid therapy. Receiver operating characteristic curves of FeNO revealed obvious differences among CVA, NAEB, and CVA/NAEB (area under the curve = 0.855, 0.699, and 0.923, respectively). The cutoff values of FeNO at 43.5 and 32.5 ppb were shown to best differentiate CVA and CVA/NAEB, respectively. FEF(25–75) was significantly lower in patients with CVA than in those with other causes. A FEF(25–75) value of 74.6% showed good sensitivity and specificity for identifying patients with CVA. CONCLUSIONS: NAEB, CVA, and UACS are common causes of chronic cough in patients with AR. FeNO can first be used to discriminate patients with CVA/NAEB, then FEF(25–75) (or combined with FeNO) can further discriminate patients with CVA from those with CVA/NAEB.
Asthma ; Bronchial Provocation Tests ; Bronchitis ; Cough ; Eosinophils ; Gastroesophageal Reflux ; Humans ; Methacholine Chloride ; Nitric Oxide ; Rhinitis, Allergic ; ROC Curve ; Sensitivity and Specificity ; Spirometry ; Sputum