Objective To investigate the drug resistance of Acinetobacter baumannii strains isolated during 2014 in a hospital of Beijing.Methods Isolated bacteria were cultured by routine method,identified and performed drug susceptibility test by bacteria a-nalysis system.Statistical analysis of Acinetobacter baumannii was conducted by Whonet5.6 software.Results A total of 226 Acin-etobacter baumannii strains were isolated from January 2014 to December 2014 in Fengtai Teaching Hospital of Capital Medical U-niversity.These strains showed the lowest resistance rates to minocycline and Cefoperazone-sulbactam(23.0% and 30.1%).The other antimicrobial resistance rates were more than 50.0%.The resistance rate of Acinetobacter baumannii varied from one depart-ment to another.Pandrug-resistant and multi-drug resistant Acinetobacter baumannii were 71.2%,1 9.0% respectively. Conclusion The resistant rates of Acinetobacter baumannii strains to a variety of antibiotics are high.It is essential to strengthen monitoring the drug resistance of Acinetobacter baumannii,as well as use antibiotics reasonable and separate patients to control Acinetobacter baumannii infection.

Objective To compare the activation of microglia in vitro induced by oligomeric and fibrillar Aβ,and research the effects of Piper Kadsura Ohwi extracts on the microglial activation.Methods Microglia were divided randomly into 4 test groups,intervened by fibrillar Aβ25-35,fibrillar Aβ25-35 + Piper Kadsura Ohwi extracts,oligomeric Aβ25-35 and oligomeric Aβ25-35 + Piper Kadsura Ohwi extracts respectively.Also a blank contrast group was set without any intervention.48 hours later,activation of microglia was tested by immunocytochemistry,using the CD68 molecule as a specific marker of microglial activation and the incidences of active microglia in different groups were compared.Results Each of the 4 test groups had a higher positive incidence of CD68 expression than that of the blank contrast (5.1％ ) (P ＜ 0.05 ) ; positive incidence of fibrillar Aβ + Piper Kadsura Ohwi extracts group (52.1％) was lower than that of the fibrillar Aβ group (60.8％) (P＜0.05) ; positive incidence of oligomeric Aβ + Piper Kadsura Ohwi extracts group (67.0％) was not significantly different with that of the oligomeric Aβ group (71.2％),P=0.101.Conclusion Both fibrillar and oligomeric Aβ has the ability to activate the silent microglia.Piper Kadsura Ohwi extracts could inhibit the activation of microglia induced by fibrillar Aβ25-35,but didn't show significant effects on the activation induced by oligomeric Aβ25-35.

Objective To explore the effect of chemokine CXCL12 and its receptor CXCR4 on proliferation,migration and invasion of epithelial ovarian cancer cells.Methods CXCR4 and CXCL12 mRNA and protein expression of human ovarian cancer cell line CAOV3 was detected by RT-PCR and immunocytochemistry.Integrin ?1 and vascular endothelial growth factor-C(VEGF-C)mRNA expression were detected in CAOV3 cells stimulated by CXCL12.The CAOV3 cells were divided into 6 groups:control group(un-stimulated),experimental group 1(stimulated by 100 ng/ml CXCL12),experimental group 2 (stimulated by 10 ng/ml CXCL12),experimental group 3(100 ng/ml CXCL12 and 10 ?g/ml neutralizing CXCR4 antibody),experimental group 4(100 ng/ml CXCL12 and 1 ?g/ml CXCR4 antagonist AMD3100),experimental group 5(10 ?g/ml neutralizing CXCR4 antibody or ascites).Methyl thiazolyl tetrazolium(MTT)was used to analyze the effects of different concentrations of CXCL12 on CAOV3 cell proliferation.Transwell invasion chamber and reconstructed basement membrane(Matrigel)were used to evaluate effect of various concentrations of CXCL12 and ascites on CAOV3 cell migration and invasion. Results CAOV3 cells expressed CXCR4 mRNA(0.70?0.10)and protein,but did not express CXCL12 mRNA or protein.Immunostaining of CXCR4 was mainly located in cytoplasm.CXCR4 mRNA was up- regulated after 100 ng/ml CXCL12 stimulation(1.24?0.14;t=-7.1088,P=0.0021).Integrin ?1 mRNA was greatly increased at 3 hours by stimulation of 100 ng/ml CXCL12(before and after stimulation 0.53?0.10,1.53?0.16;P0.05).Experimental group 1 stimulated the migration and invasion of CAOV3 cells in chemotaxis assay compared with control group and experimental group 2(number of cell migration respectively 523.3?25.2,108.0?7.2,211.7 ?24.7,number of cell invasion respectively 39.3?4.0,4.0?1.0,15.7?3.1;P

Objectives To explore the impact of Aβ oligomer(Aβo) and fibrillar Aβ( Aβf) on the ethology of model rat and to explore whether or not piper kadsura ohwi(PKO) can ameliorate the ethological changes.Methods AD animal model was made by continuous intracerebroventricular infusion of Aβ through mini-osmotic pump.After surgery,rats of Aβo + PKO group and Aβf + PKO group received intraperitoneal injection of 10％PKO everyday,rats from Aβo + DMSO group and Aβf + DMSO group received intraperitoneal injection of 2.5％ DMSO.The treatment lasted 5 weeks.Morris water maze experiment began on the 31st day after surgery.Results ( 1 ) Acquisition trials:the escape latency was longer in Aβo group ( ( 89.40 ± 7.20 ) s ) than that in Aβf group ( (65.00 ± 10.89 ) s ) ; escape latency was shorter in Aβf + PKO group ( ( 34.00 ± 11.26 ) s) than that in Aβf group and Aβf + DMSO group( (60.6 ± 6.95 ) s) ; escape latency was shorter in Aβ3o + PKO group ( ( 65.33 ±8.89) s) than that in Aβo group and Aβo + DMSO group ( ( 85.60 ± 6.02) s).( 2 ) Robe trial:the times ( 3.00 ±0.71 ) and the proportion of time (0.23 ± 0.02 ) and path(0.23 ± 0.04) spent in the target quadrant in Aβo group was less than that in Aβf group(6.00 ± 1.58,0.25 ± 0.01,0.26 ± 0.03 ) ; the three parameters in Aβf + PKOgroup were more than those in Aβ3f group and Aβf+ DMSO group; the three parameters in Aβo + PKO group were more than those in Aβo group and Aβo + DMSO group.Conclusions Aβ oligomer had a more severe impact on the ethology of AD model rats than fibrillar Aβ did; piper kadsura ohwi may ameliorate the ethological changes of AD model rats.

Objective To evaluate the association of expressions of leptin and leptin receptor in breast cancer with tumorigenesis and development and clinicopathologic factors.Methods The expression of leptin and leptin receptor was performed in 132 cases of breast cancer,66 cases of benign breast lesions,and 30 cases of corresponding normal breast tissue by using immunohistochemical methods. ResultsThe expressions of leptin and leptin receptor were 76.5 ％ (101/132) and 70.5 ％ (92/132).It was significantly higher than that in benign breast lesions 56.1％ (37/66) and 56.1％ (37/66),and corresponding normal breast tissues 46.7 ％ (14/30) and 43.3 ％ (13/30) (x2 =8.72,P =0.003,x2 =4.04,P =0.044,x2 =10.57,P =0.001and x2 =7.94, P =0.005).The level of leptin expression showed a significant correlation with the level of leptin receptor (r =0.307,P ＜ 0.05).The expression of leptin and leptin receptor in breast cancer tissue were not significantly related to ages,menopause,tumor size,classification,pathological type,distant metastasis,ER and PR expression (P ＞ 0.05).The positive rate of leptin in patients with lymph node metastasis was 91.7 ％,which was significantly higher than that in patients without lymph node metastasis (67.9 ％,x2 =10.65,P =0.002).Conclusions The expressions of leptin and leptin receptor have a significant correlation with breast cancer and may have the promoting effect on the carcinogenesis and development of breast cancer.

Objective To observe the activation of anti-viral innate immune response of type Ⅰinterferon and inhibition of hepatitis B virus (HBV)genome replication in mice by HBV-3p-siRNA. Methods HBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA (NC-siRNA)and non-modified HBV-siRNA were used as control groups.Blood samples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group.The model group was only injected with pGL3.0-HBV1 .2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3p-siRNA group received peritoneal injection of HBV-3p-siRNA.The interferon-β(IFN-β)and hepatitis B surface antigen (HBsAg)in serum were detected by enzyme linked immunosorbent assay (ELISA).The copies of HBV DNA were assessed by fluore scence quantitative polymerase chain reaction (PCR ).The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Results Serum level of IFN-β was (12.37±5 .32)pg/mL in model group,(22.61 ±6.29 )pg/mL in negative control group,(26.40±5 .39)pg/mL in HBV-siRNA group and (68.37± 21 .00 ) pg/mL in HBV-3p-siRNA group.The secretions of IFN-β into serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group (F =23.988 and 46.523,respectively,both P <0.01).Serum level of HBsAg was (2 864.86±907.11 )ng/mL in model group,(2 198.86±456.89 )ng/mL in negative control group,(1 049.71 ± 396.28 )ng/mL in HBV-siRNA group and (640.86±383.08)ng/mL in HBV-3p-siRNA group.The expressions of HBsAg were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F = 23.537 and 39.144, respectively;P =0.025 and 0.010,respectively).Serum level of HBV DNA was (2.54 ×104 ±1 .46 × 104 )copy/mL in model group,(2.22×104 ±2.62×103 )copy/mL in negative control group,(3.59×103 ±2.88×103 )copy/mL in HBV-siRNA group and (2.65 ×103 ±1 .46×103 )copy/mL in HBV-3p-siRNA group.Serum level of HBV DNA were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F =15 .013 and 16.741 ,respectively,both P <0.05 ).All of the indicated siRNA used in the experiments showed no apparent effects on the body mass index of the mice models.Conclusion HBV-3p-siRNA,which induces the production of IFN-β and inhibits HBV replication through gene silencing in vivo ,may be a powerful bifunctional antiviral molecule.

The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.

In this study,nearly 200 endophytic bacteria were isolated from different part of Huperzia serrata, over 60 bacterium with clear antifungal activity were selected from those cultures.Among them,strain H-6 exhibited the highest antifungal activity which was strongly inhibits the growth of many plant pathogenic fungi such as Sclerotinia scleroliorum,Fusarium graminearumt,Sclerotinia libertiana,Phytophthora capsici Leonia and Sesame fusarium wilt.According to the characteristics of morphology,physiology and biochem- istry tests and the comparison of 16S rDNA sequence,the strain H-6 was similar to the Burkholderia.So strain H-6 was identified as Burkholderia sp.H-6.The results also showed that Burkholderia sp.H-6 was markedly different from Burkholderia cepacia that was applied widely in agriculture as antagonistic bacteria. The medium and culture conditions of the strain all were optimized by single factor and orthogonal experiments.In the investigation of the culture condition,growth was carried out in a basal medium(potato juice)and gradually supplemented with the various ingredients to be investigated.The major ingredients be- ing investigated included carbon sources and nitrogen sources.The optimal antifungal activity production condition is growth in a medium(potato juice with 2.5%mannitol and 0.1%NaNO3),initial pH 4.0 at 28℃.

AIM To determine the dynamic changes of monoamine neurotransmitters and their metabolites in various brain regions of cerebral ischemia reperfusion mice. METHODS Concentrations of monoamine neurotransmitters such as norepinephrine (NE), dopamine (DA), serotonin (5-HT) and metabolites were determined by HPLC-ECD on d 0,1,3,5 and d 20 after cerebral ischemia reperfusion by common carotid artery occlusion. RESULTS The cerebral ischemia reperfusion mice showed decreased concentrations of NE, MHPG, DA, DOPAC, 5-HT and 5-HIAA in various brain regions, especially in hippocampus. CONCLUSION Several neuron systems play an important role in neurons damage of cerebral ischemia reperfusion, especially the NE and DA in hippocampus which is sensitive to the ischemia damage. The data offer useful guides for clinical treatments of cerebral ischemia diseases.

AIM:To observe the effect of fresh amniotic membrane transplantation ( FAMT) in acute ocular chemical burns.
●METHODS:A prospective study of 25 consecutive cases (36 eyes) with acute ocular chemical burns were treated with FAMT. The clinical efficacy was observed such as the time of amniotic membrane absorbed, corneal epithelialization & transparency, visual acuities and complications.
●RESULTS: With follow-up ranged from 3 to 6mo, 31 eyes′ amniotic membrane were dissolved in 2wk (86%). A total of 33 eyes showed corneal epithelialization in 4wk ( 92%) , 3 eyes showed persistent corneal epithelial defects and need secondary limbal stem cell transplantation or corneal transplantation ( 8%) . A total of 10 eyes showed superficial corneal vascularization (28%), 6 eyes′ cornea were opacity in part (17%), and one eye was symblepharon (3%).
●CONCLUSION:Early FAMT is an effective treatment in the management of acute ocular chemical burns to support epithelial healing, restore ocular surface integrity with potential to improve vision and reduce the incidence of complications. Furthermore, FAMT has advantages of easily obtain and convenient usage, which is suitable in local hospital of our country.