Objective To observe the effects of treating knee osteoarthritis(KOA)combined with synovitis with Gubiqing,and discuss its mechanism.Methods A total of 60 cases with KOA were randomly recruited into a control group and a treatment group,30 cases in each.TCM symptoms,signs and health assessment questionnaire(HAQ)were observed before and after the treatment.Results The tohal therapeutic effect was 90%and 70%in the treatment group and the control group respectively.There was significant difference between the two group(χ~2=48,P=0.003).Body signs and HAQ were also greatly improved in the treatment group(t=0.004、P=0.008).Conclusion Gubiqing can not only restrain chondrocyte apoptosis in knee osteoarthritis combined with synovitis,but also relive the damage of articular cartilage.

Objective To explore the therapeutic effect, mechanism and safety of Qingrehuashi recipe in treating chronic superficial gastritis with erosion pertaining to spleen-stomach damp-heat syndrome. Methods 84 cases of chronic superficial gastritis with erosion were recruited into a control group and a treatment group randomly, with each groups containing 42 cases. The treatment group was treated with Qingrehuashi recipe, while the control group was treated with Qingweizhitong granule. Both groups were treated for 8 weeks. The clinical manifestations and signs, gastroscopic results and pathological effects were observed before and after the treatment. Results The total effective rate and curative rate in the treatment group were superior to those in the control group (P<0.05) . Conclusion Qingre Huashi recipe is effective and safe in in treating chronic superficial gastritis with erosion pertaining to spleen-stomach damp-heat syndrome. The mechanism might be related to its effects of anti-inflammation, anti-oxidation, eradicating Hp, promoting gastric motility and enhancing gastric mucosal barrier.

Objective To explone the therapeutic effect and mechanism of Quyuhuatan decoction in treating type 2 diabetes mellitus combined with abnormal metabolism of lipid. Methods 85 cases of type 2 diabetes mellitus combined with abnormal metabolism of lipid were recruited into a control group and a treatment group randomly. The treatment group (45cases) was treated with Quyuhuatan decoction and subcutaneous insulin injection while the control group(40cases) was treated with subcutaneous insulin injection exclusively. Both groups received the treatment for for 8 weeks. The result of TC, TG, LDL-C, and HDT-C were observed before and after the treatment. Results The improvement of blood lipid in the treatment group was superior to that in the control group (t=0.011, P<0.05) . Conclusion Quyuhuatan decoction is effective in treating type 2 diabetes mellitus combined with abnormal metabolism of lipid. The mechanism might be related with its cutting off pathological paths of hyperlipidemia.

Bacterial extracellular metalloproteases ( BEMPs) are a large group of metal ion-contai-ning proteases. All BEMPs identified so far are endopeptidase or endoprotease. BEMPs can be classified into nine metalloprotease families based on the sequences and structures of enzymatic molecules. Double-valence zinc ion ( Zn2+) is necessarily required by catalytic centers of most BEMPs. The main function of BEMPs in non-pathogenic heterotrophic bacteria is to hydrolyze environmental proteins and polypeptides to provide vari-ous amino acids as nutrients. However, BEMPs of pathogenic bacteria, serving as important virulence fac-tors, help the pathogens invade into hosts and spread in hosts. In recent years, the roles and mechanism of BEMPs in bacterial pathogenesis have attracted great attention. Here, we make a brief review about the structures and types as well as the functions and pathogenic roles of BEMPs.

Objective To clone and express the new gene R049 of uropathogenic Escherichia coli 132.and to investigate the immunopmtective effects of the R049 recombinant protein on mice.Methods The pmkaryotic expression system of gene R049 was constructed by directed cloning.Thereafter,the R049 recombinant protein Was expressed and purified by Ni affinity chromatography.Polyclonal antibody was pre-pared by immunizing BALB/c mice with R049 recombinant protein.The R049 recombinant protein and whole bacterial proteins of UPEC132 were analyzed by SDS-PAGE and Western blot.BALB/c mice were im-munized with R049 recombinant protein before challenged by UPECl32 through urinary tract.Then the differences of urine and renal colony counts between immunization group and control group were compared.Results The recombinant strain E coli BL21(DE3)/pET32a-R049 ORF was constructed successfully,and the relative molecular mass of the R049 recombinant protein was 66.9×103 and its purity was up to 95% af-ter purification.The titer of polyclonal antibody wag≥1:102 400 analyzed by indirect ELISA.Both of the R049 recombinant protein and whole bacterial proteins of UPECl 32 were confirmed to show specffic reactions on the antiserunl throughh Western blot.The animal experiments showed the urine and renal colony counts of immunization group were significantly lower than that of the control group(P<0.01,P<0.05).Conclu-sion The new gene R049 of uropathogenic E.coli 132 had immunopmtective effects on mice and the defini-tive mechanism would be needed to further study.

Objective To investigate the genotypes of extended spectrum β-lactamases (ESBLs) and their carrying modes in Escherichia coli (E.coli) isolates,and to analyze the mechanism of protein phosphorylation and ESBLs gene expression induced by β-lactam antibiotics or inhibited by histidine kinase inhibitors.Methods The predominant genotypes of ESBLs (KPC,TEM,SHV and CTX-M) and their carrying modes were identified by PCR and sequencing analysis.E-test and micro-tube dilution method were applied to measure minimal inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs).Immobilized metal ion affinity chromatography,bacterial protein phosphorylation detection kit and real-time fluorescent quantitation RT-PCR were performed to analyze the enhancing effects of 1/4 MIC penicillin or cefotaxime or the inhibitory effects of histidine kinase inhibitors (closantel,bromized or iodized methylimidazol) on protein phosphorylation and the expression of ESBLs at mRNA level in E.coli isolates.Results In 183 β-lactam antibiotics-resistant E.coli isolates,TEM and CTX-M genes (83.1％ and 77.1％) were highly expressed than other two ESBLs genes with a prevalent carrying mode of coexisting (65.0％) (P＜0.05).Penicillin or cefotaxime at 1/4 MIC induced the protein phosphorylation and promoted the expression of TEM,SHV and CTX-M at mRNA level (P＜0.05).Closantel (200 μmol),bromized methylimidazol (2 or 10 μmol) or iodized methylimidazol (20 or 50 μmol) could neither kill E.coli isolates nor inhibit their growth,but could inhibit the protein phosphorylation induced by above mentioned antibiotics and enhance the expression of ESBLs at mRNA level (P＜0.05).Moreover,the susceptibility of antibioticresistant E.coli strains to penicillin and cefotaxime were increased (P＜0.05).Conclusion TEM and CTX-M were the predominant genotypes of ESBLs carried by β-lactam antibiotics-resistant E.coli strains isolated from Zhejiang province,which were mostly found in a TEM plus CTX-M carrying mode.Sublethal dose of β-lactam antibiotics could up-regulate the expression of ESBLs genes in E.coli isolates via TCSS,but it could be inhibited by histidine kinase inhibitors.

Objective To investigate the role of pathogenic Leptospira interrogans lipopolysaccha-ride (L-LPS) and outer membrane proteins (L-OMP) in the apoptosis of mouse macrophages (J774A.1) and their association with Fas/FasL pathway .Methods Phenol-water extraction and Triton X-114 phase separation were used to extract L-LPS and L-OMP from L.interrogans serogroup icterohaemorrhagiae serovar Lai strain Lai, respectively.Polymyxin B ( PMB) and protease K ( PK) were used to treat L-LPS and L-OMP, respectively.J774A.1 cells were stimulated by L.interrogans strain Lai with or without ultraviolet inactivation.In parallel, the cells were stimulated by extracted L-LPS and L-OMP with or without PMB and PK treatments .The apoptosis and necrosis of J 774 A.1 cells before and after treatment were detected by flow cytometry.The siRNAs were used to silence the expression of Fas or FasL gene in J 774A.1 cells and their inhibitory effects were further validated by using real-time fluorescent quantitative RT-PCR.Flow cytometry was used to detect the effects of L-LPS or L-OMP on the apoptosis of J774A.1 cells with Fas or FasL gene-knockdown .Results L.interrogans strain Lai with or without ultraviolet inactivation could cause similar early apoptosis rates (47.1%and 55.6%) and late apoptosis/necrosis rates (7.6%and 7.9%).The ear-ly apoptosis rates of 1×105 J774A.1 cells were 40.4%and 34.0%after the treatment with 100 ng of L-LPS and 100 μg of L-OMP for 4 h.The late apoptosis/necrosis rates of the cells were 7.5%and 6.9%upon the treatments with L-LPS and L-OMP, respectively.However, the apoptosis or necrosis of the cells was not ob-served when using L-LPS and L-OMP pre-treated by PMB and PK, respectively.Silenced expression of Fas or FasL gene reduced the L-LPS-induced J774A.1 cells apoptosis (P<0.05), while decreased early apopto-sis rate of J774A.1 cells mediated by L-OMP was only observed in Fas gene-knockdown cells (P<0.05). Conclusion Both L-LPS and L-OMP can cause the Fas/FasL-associated apoptosis of macrophages , which is beneficial for L.interrogans to establish the productive infection in hosts .

We isolated and identified the bacterial pathogen in a pyogenic balanoposthitis patient and investigated the drug resistance and its mechanism of the isolate.Urethral secretions and balanus pustule liquids were collected for microscopic examination after Gram-staining and detection of mycoplasma using Mycoplasma IST 2 kit.The two samples were inoculated on Columbia blood plate,N.gonorrhoeae selective plate and chromID Candida plate for isolation.The obtained colonies were identified by VITEK 2-compact automatic bacterial detection and analysis system.Moreover,PCR was performed to detect 16S rRNA gene of N.gonorrhoeae in the samples and colonies.KB method was applied for detecting susceptibility of five common antibiotics against the isolate.The β-lactamase and extended spectrum β-lactamase confirmatory tests were used to investigate the enzyme production of the isolate as well as drug resistance-associated tetM,TEM,mefA and ermF genes in the isolate were detected by PCR.Results showed that all the clinic samples showed negative for mycoplasma.All the isolating cultivation results of urethral secretions were negative while the balanus pustule liquids provided positive isolating cultivation in the blood and selective plates.The VITEK 2-compact system and 16S rRNA-PCR revealed that the isolated strain belongs to N.gonorrhoeae.The isolate can produce β-lactamases and resist to penicillin G,ciprofloxacin and tetracycline.The tetM,TEM,mefA and ermF genes could be found in the isolate's genome.The patient's balanoposthitis is caused by infection of N.gonorrhoeae.The multidrug resistance of Neisseria gonorrhoeae isolate is closely associated with its carried resistant genes.

Objective To explore the impact of atrial fibrillation (AF) recognized at primary diagnosis on clinical features and outcomes of patients with AF in emergency service.Methods Data were collected from consecutive patients admitted in resuscitation room in the Department of Emergency (ED) of a major comprehensive teaching hospital,from January 1,2011 through December 31,2015.Patients were checked by electrocardiogram examination and / or monitored in resuscitation room after admission,and were divided into patients with AF recognized at a primary diagnosis and those with AF judged by alternative primary diagnoses in ED.The main criteria of prognosis were the length of resuscitation room stay,number of repeated ED visits,and outcome scale (such as death,transferred to intensive units,transferred to general wards,or direct discharge).Non-paired student t test,x2,and circular distribution analysis were performed using SPSS 10.0 and EXCEL 2007 software.Results A total of 929 patients with mean age of (70.3 ± 12.7) years,and 502 (54.0％) female were enrolled.There were 122 cases with AF not recognized at primary diagnosis but by an alternative primary diagnosis (non-primary group,NPG),and 807 cases with AF recognized at primary diagnosis (primary group,PG).Compared with the PG,the patients were older [(76.9 ±9.3) vs.(68.7 ± 14.4),P ＜0.01],had more comorbidities [(1.75 ± 1.26) vs.(0.08±0.39),P＜0.01],higher APACHE Ⅱ scores [(17.89±8.19) vs.(8.64±4.15),P＜ 0.01],longer resuscitation room stay (P ＜ 0.01),higher mortality (11.5％ vs.0.2％,OR =52.176,95％ CI:11.698-232.710,x2 =78.928,P ＜ 0.01) and a higher percentage of transferring to intensive careunit (14.8％ vs.5.1％,OR=3.234,95％CI:1.791-5.838,x2 =16.674,P＜0.01) in NPG.There were no significant difference in number of repeated-visits in ED between the PG and the NPG.Conclusion Patients with AF in the ED judged by alternative primary diagnosis are older and have more comorbidities,higher mortality and higher probability to be transferred to intensive care unit than patients with AF directly recognized by a primary diagnosis.This cohort of patients with special characteristics should be meticulously cared for and be distinguished from the patients with AF crystal clear at a primary diagnosis.Future studies are needed to examine the specific impact of AF on outcomes in the setting of primarydiagnoses in ED.

Objective To study the distribution of the specific fragment R049 of uropathogenic E. coli(UPEC) 132 in UPEC and fecal E. coli strains. Methods The specific fragment R049 was amplified by PCR from 20 UPEC strains and 40 fecal E. coli strains, and 5 genes encoding virulence factors (papC, fimH, hly, aer, cnf1) were detected from fragment R049 positive strains. Pulse field gel electronphoresis (PFGE) was applied for isolating the Xba Ⅰ restriction fragments of the genomes of fragment R049 positive strains, and then Southern blot was applied for analyzing the distribution features of the positive hybridization bands by digoxin-labeled R049 ORF probe. Results The specific fragment R049 was amplified from 8 of 20 UPEC strains (40%) and 3 of 40 fecal E. coli strains (7.5%), and statistics analysis showed significant difference (P<0.01). The specific fragment 11049 was closely related with 5 virulence factors of UPEC in the fragment R049 positive strains. Southern blot showed the sizes of positive bands were 150 kb, 15 kb and 240 kb in 3 fecal E. coli strains, 350 kb in 6 of 8 UPEC strains, and 280 kb and 25 kb in the rest two UPEC strains. Conclusion The specific fragment R049 of UPEC132 possessed the feature of clustering distribu-tion in domestic isolated UPEC strains.