Objective To understand the homology of clinical isolates from patients with carbapenem-resistant Klebsiella pneumoniae(CRKP)infection and isolates from environment in a medical institution.Methods One CRKP strain isolated from a patient in this hospital and 4 strains of Klebsiella pneumoniae(K.pneumoniae)isolated from patient's surroundings were collected,susceptibility of 5 strains to commonly used antimicrobial agents was detected,production of carbapenems in 5 strains were detected by modified Hodge testing and carbapenem inactivation method(CIM),homology analysis was performed by pulsed-field gel electrophoresis(PFGE).Results Antimicrobial susceptibility testing results showed that 5 strains of K.pneumoniae(1 from patient,4 from the patient's ward surroundings,including hands of nursing aides,solution bottle opening,handle for lifting and dropping bed,and bedrail)were all resistant to other antimicrobial agents except to cephamycin and aminoglycosides.The modified Hodge testing and CIM confirmed that 5 strains all produced carbapenemases;PFGE results showed that electrophoretogram of CRKP isolated from solution bottle opening of ward,bedrail,and handle for lifting and dropping bed were the same as CRKP isolated from patient,while electrophoretogram of CRKP isolated from hands of nursing aides had 2 different bands,there was a close relationship between the strains.Conclusion The same type of CRKP were isolated from patient and his surroundings,it is necessary to implement healthcare-associated infection(HAI)control system,isolate infected patient,and strengthen environmental cleaning and disinfection,so as to avoid the outbreak of HAI.

Objective To evaluate the correlation between pathological findings and clinical characteristics of patients with Sj(o)gren's syndrome (SS). Methods Eighty-four patients diagnosed with SS from 119 suspected ones at Peking University First Hospital were recruited. According to the pathological changes of lip biopsy, 64 patients were focus score (FS)( + ) and 20 patients were FS (-). In the FS( + ) group, 20 patients had germinal center (GC)( + ) and 44 patients had no GC. x2 test and t test were used for statistical analysis. Results The duration of symptoms of dry eyes or dry mouth in the FS (+) group was longer than that in the FS (-) group (P＜0.05). In the FS ( + ) patients, those with GC (+) had significantly longer duration of xerothalmia or xerostomia, higher serum IgG levels, greater FS score, and higher incidence of system involvement than those without GC patients (P＜0.05). In the FS (-) group, lip biopsies showed degenerative changes in 6 (30%) patients. Those with degenerative changes had longer duration of dry eyes and dry mouth and higher incidence of system involvement. Conclusion GC (+) in FS (+) patients and degenerative changes in FS (-) patients from lip biopsy are associated with severer clinical manifestations in patients with SS, suggesting that more clinical attention should be paid to this subgroup of patients.

Objective To analyze pathogenic bacteria distribution and antimicrobial susceptibility in children with acute otitis media(AOM ) .Methods Otorrhea samples from 146 episodes of AOM were cultured .The antimi‐crobial susceptibility of the main pathogenic bacteria was determined .The results were analyzed by SPSS19 .0 .Re‐sults 1) The strains of bacteria were isolated from 109 children with the positive rate of 74 .66% .Streptococcus pneumoniae (SP ) was the major bacteria(64 episodes ,58 .72% ) ,followed by staphlococcus aureus(SA) (19 epi‐sodes ,17 .43% ) .2) Sp was all sensitive to vancomycin ,levofloxacin ,moderate to penicillin ,amoxicillin ,cefo‐taxime ,and highly resistent to erythromycin and clindamycin .Staphlococcus aureus were all sensitive to vancomy‐cin ,tetracycline ,and Amy card ,and moderate to amoxicillin clavulanic acid potassium ,cefoxitin ,and oxacillin ,all resistent to penicillin and ampicillin .3) The strains of SP in age≤1year ,>1 -3years ,and >3 years respectively were 31(50 .82% ) ,25(56 .82% ) ,8 (19 .51% ) .There were significant differences between them(χ2 =14 .073 ,P=0 .001) .4)The strains of SP in 2012 ,2013 ,2014 respectively were 16(30 .19% ) ,22(48 .89% ) ,26(54 .17% ) ,There were significant differences between them(χ2 =6 .557 ,P=0 .038) .The antimicrobial susceptibility of SP had no sig‐nificant differences among 2012 ,2013 ,2014 ,but a yearly resistance decreasing trend was seen .Conclusion SP was the main bacterial contributor for AOM in Wuhan children .SP detection rate increases every year ,mainly in chil‐dren less than 3 years old .T he antimicrobial susceptibility is stable .

A polydimethylsiloxane ( PDMS )/glass microfluidic chip consisting of a three-layer sandwich structure and three-parallel micro-electrode system was fabricated for the separation of polystyrene microspheres according to the particle size in high conductive solution by electric field produced by alternating current. The principle of electrokinetics of microspheres directional movement was investigated. The results showed that, when the applied voltage was 14 V at 100 kHz, the separation efficiency of 10 and 25 μm polystyrene microspheres was the best. Similarly, with a voltage of 10 V at 2 MHz, the separation efficiency of 5 and 25 μm polystyrene microspheres could achieve the highest value. Meanwhile, the voltage of 11 V at 1 MHz was suitable for the separation of 5 , 10 and 25 μm polystyrene microspheres with the separation efficiency of over 90%. At the same time, the formation of the laminar region in the middle of the electrode gap was the key role of microsphere separation.

ABSTRACT:Nested‐PCR and loop‐mediated isothermal amplification (LAMP) were applied to identify the Borrelia burg‐dor f eri (B .burgdor f eri) in ticks in this study .A total of 112 adult ixodes were collected from vegetation in a forest area and farm animals in Xunhua County ,Qinghai Province and Xinbin County ,Liaoning Province .The ticks were examined for the presence of B .burgdorferi by nested‐PCR and LAMP .Results showed that 12 in 51 samples were found positive in Xunhua County (23 .53% ) .While positive rate in Xinbin County was 29 .51% with 18 samples positive in 61 samples .In total of 112 tick samples ,the PCR‐positive rate was 17 .86% with 20 positive samples identified ,whereas 15 positive samples were con‐firmed with positive rate of 13 .39 % by LAMP assay .There was no significant difference between the two assays (Х2 =0 .85 , P>0 .05) .Results suggest that both nested‐PCR and LAMP could be used in identifying B .burgdorferi in ticks .Combina‐tion of these two assays could improve the testing results .This is the first report of B .burgdorferi in ticks in Xunhua and Xinbin counties ,and helps to complete the database of the infection rate of B .burgdor f eri in ticks in the widely‐forested area of China .

Objective To investigate the protective effect of rhein lysinate (RHL)on the liver of the models with diabetic rats,and to provide basis for research on treatment of fatty liver in the patients with diabetes mellitus. Methods The models of diabetic rats were established by intraperitoneally injecting streptozotocin(STZ).40 rats were divided into control,model,25 mg·kg-1 RHL,and 50 mg·kg-1 RHL groups(n=10).The levels of malonaldehyde (MDA)and the activities of superoxide dismutase (SOD)and glutathione peroxidase (GSH-Px) were detected by thiobarbituric acid method, pyrogallol autoxidation method, and NADPH coupling method, respectively.The pathological changes of liver tissue were observed by hematoxylin and eosin (HE)staining;the content of fat in liver tissue was observed by Nile red staining;the expression levels of fat synthesis-related proteins were detected by Western blotting method.Results Compared with control group,the body weight of the rats in model group was decreased and the levels of blood glucose,total cholesterol(TC)and triglyceride(TG)were increased (P<0.05);the activities of SOD and GSH-Px in liver tissue were decreased (P<0.05);there were a plenty of fat vacuoles and fat accumulation in liver tissue. The signal pathway of fat synthesis-related ERK1/2-SREBP-1c was activated in model group;compared with model group,it was inhibited in 25 and 50 mg· kg-1 RHL groups (P<0.05).Compared with model group,the blood glucose,TC and TG of the rats in 25 and 50 mg ·kg-1 RHL groups were decreased (P<0.05);the activities of SOD and GSH-Px were increased (P<0.05);however the body weight had no change. Compared with model group, the fatty vacuoles and the fatty accumulation of liver tissue in 25 and 50 mg·kg-1 RHL groups were decreased. Conclusion The hepatic protection of RHL is correlated with the inhibition of oxidative stress, fat degeneration and fatty accumulation of liver tissues.

Objective:To evaluate the role of nuclear receptor subfamily 1, group D, member 1/brain and muscle Arnt-like 1(Rev-erbα/Bmal1) signaling pathway in myocardial ischemia-reperfusion (I/R) injury in diabetic rats and its relationship with autophagy.Methods:SPF-grade adult male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-220 g, were used in this study.Type I diabetes mellitus was induced by intraperitoneal streptozotocin 60 mg/kg.The rats were continuously fed for 8 weeks after successful establishment of the model.Thirty rats with type 1 diabetes mellitus were divided into 3 groups by a random number table method: diabetic sham operation group (DS group, n=6), diabetic myocardial I/R group (DI/R group, n=12) and diabetic myocardial I/R plus Rev-erbα antagonist SR-8278 group (DI/R+ SR group, n=12). Eighteen non-diabetic rats were divided into 2 groups by a random number table method: non-diabetic sham operation group (NS group, n=6) and non-diabetic myocardial I/R group (NI/R group, n=12). The myocardial I/R model was established by ligation of the left anterior descending coronary artery for 30 min followed by 120-min reperfusion in anesthetized rats.SR-8278 2 mg/kg was intravenously injected via the femoral vein at 1 h before ischemia in group DI/R+ SR.Blood samples were collected from the carotid artery immediately after the end of reperfusion for determination of serum troponin I (cTnI), creatine kinase-MB (CK-MB) and lactic dehydrogenase (LDH) levels (by enzyme-linked immunosorbent assay). Then the rats were sacrificed, and myocardial tissues were obtained for determination of myocardial infarct size (by TTC method), expression of Rev-erbα and Bmal1 mRNA (by real-time polymerase chain reaction) and expression of Rev-erbα, Bmal1, microtubule-associated protein 1 light chain (LC3) Ⅱ and LC3Ⅰ (by Western blot) and for calculation of the ratio of LC3 Ⅱ/LC3Ⅰand the number of autophagosomes (with a transmission electron microscope). Results:Compared with group NS, the percentage of myocardial infarct size, serum levels of cTnI, CK-MB and LDH and the number of autophagosomes were significantly increased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas increased in group NI/R, and serum levels of cTnI, CK-MB and LDH were increased, the number of autophagosomes was decreased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated and the ratio of LC3 Ⅱ/LC3Ⅰwas decreased in group DS ( P<0.05). Compared with group NI/R, the percentage of myocardial infarct size and serum levels of cTnI, CK-MB and LDH were significantly increased, the number of autophagosomes was decreased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas decreased in group DI/R ( P<0.05). Compared with group DS, the percentage of myocardial infarct size, serum levels of cTnI, CK-MB and LDH and the number of autophagosomes were were significantly increased, the expression of Rev-erbα and its mRNA in myocardial tissues was up-regulated, the expression of Bmall and its mRNA was down-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas increased in group DI/R ( P<0.05). Compared with group DI/R, the percentage of myocardial infarct size, serum levels of cTnI, CK-MB and LDH and the number of autophagosomes were significantly decreased, the expression of Rev-erbα and its mRNA in myocardial tissues was down-regulated, the expression of Bmall and its mRNA was up-regulated, and the ratio of LC3 Ⅱ/LC3Ⅰwas increased in group DI/R+ SR ( P<0.05). Conclusion:Rev-erbα/BMAL1 signaling pathway is involved in the process of myocardial I/R injury by regulating cell autophagy in diabetic rats.

Objective To investigate the cytotoxicity of γδ T cells to HIV-1 latency cells in patients with early HIV-1 infection.Methods Sixteen early HIV-1-infected patients were enrolled in this study.Peripheral blood mononuclear cells (PBMCs) of patients were isolated and γδ T cells were expanded using zoledronate (5 μmol/L) and interleukin (IL)-2 (1 000 IU/mL) ex vivo.Lactic dehydrogenase (LDH) was used to detect the cytotoxic role of γδ T cells to HIV-1 latency cells(J-Lat Full Length Clonel0.6).The phenotype of γδ T cells before and after expansion and the intensity of GFP in HIV-1 latency cells were detected by flow cytometry.Results Zoledronate plus IL-2 stimulated rapid and large γδ T cells proliferation ex vivo (P＜0.001).γδ T cells showed high cytotoxici ty to latency cells,and the intensity of GFP in latency cells was decreased significantly (P＜0.05).Moreover,expanded γδ T cells displayed cytotoxic NK-like phenotype,the frequency of CD56+ Vδ2 T cells in patients with early HIV-1 infection was significantly higher than that of healthy controls.Conclusions γδ T cell has an ability to eradicate HIV-1 latency,and γδ T cell-based autologous or xenogenous adoptive immunotherapy will have promise prospects to cure HIV-1 infection.

Objective To investigate the change of intracellular calcium ion concentration in prostate smooth muscle cells of SD rats with chronic abacterial prostatitis under high potassium solution.Methods SD rats were divided into experiment group and control group.The CP model was set up by castration and estradiol injection.The PSMC was cultured and purified in vitro.Laser confocal scanning microscope was used after the ceils were incubated with Quest Fluo-8TM.The cells were treated with high potassium solution,and the change of fluorescence intensity was observed.Results The pathologic specimens of the experiment group showed typical pathologic characteristics of chronic prostatitis under light microscope,the control group without inflammation performance.Using immunocytochemistry method confirmed that the experiment group and the control group were prostate smooth muscle cells.The change of fluorescence intensity of [Ca2+] i in the experiment group and control group in the high potassium solution was 27.86 ± 9.88 and 7.61 ± 4.31.There were statistically significant differences between the two groups (P ＜ 0.01).Conclusions High potassium solution cause intracellular calcium ion concentration increased.

Objective To understand the pattern of and influencing factors on leisure time physical exercises among adults in Zhejiang. Method Totally 17 437 residents aged 18 years and over selected by stratified cluster random sampling in Zhejiang in 2010 were investigated. Result Overall 21.82%(95%CI:17.12%-27.38%) of adults took part in leisure time physical exercises in 2010. The proportion was higher in urban (29.04%,95%CI:23.46%-35.33%) than in rural (18.81%,95%CI:12.89%-26.61%) (χ2=222.06, P<0.05) areas, and also higher in females (22.91%,95%CI:17.66%-29.15%) than in males (20.56%,95%CI:16.39%-25.48%) (χ2=13.94, P<0.05). Adults aged 18-24 and 55-74 years were more likely to take apart in physical exercises (χ2=266.73, P<0.05), and the lowest proportion was found among farmers (5.33%,95%CI:3.99%-7.11%) (χ2=2 078.40, P<0.05). These proportions both increased along with education level and family income increase (χ2=444.87, P<0.05;χ2=332.20, P<0.05). Overall 83.04%and 90.08%of physical exercisers took part in exercises at least 3 times per week and 30 minutes per time, only 30.67% of those reached moderate or vigorous intensity, and overall 5.38% (95%CI: 3.85%-7.48%) of adults took part in regular leisure time physical exercises in 2010. Multiple logistic regression showed that age, education level, occupation and chronic disease history have influence on regular physical exercises. Conclusion The leisure time physical exercises level was generally low among adults in Zhejiang. Young and middle-aged adults with less education and low income especially farmers should be put on emphasis.