Objective To analyze the clinical characteristics of acinetobacter baumannii infection and drug resistance tendency of our hospital in 2014 ,so as to promote the clinical rational use of antibiotics .Methods The study was a retrospective review ,the re‐sults of clinical distribution and drug resistance of acinetobacter baumannii isolated from our hospital in 2014 were analyzed .The an‐timicrobial susceptibility testing(AST) of acinetobacter baumannii was determined by K‐B disk diffusion method and minimal inhib‐itory concentration(MIC) test ,respectively .The AST was performed as recommended by CLSI 2010 .Results A total of 299 strains of acinetobacter baumannii were isolated from clinical specimens throughout the year .Of the 299 Acinetobacter baumannii isolates , 268 strains(89 .63% ) were isolated from sputum ,165 strains(55 .18% ) of Acinetobacter baumannii were from intensive care unit (ICU) and 52 strains(17 .39% ) were from neurosurgery .The resistance rate of Acinetobacter baumannii to cefoperazone/sulbactam was 5 .02% while its resistance to Imipenem and Meropenem significantly increased to 52 .17% and 56 .86% ,respectively .And the resistance rates ofβ‐lactams ,fluoroquinolones and aminoglycosides were higher than 60% .Conclusion The isolation rate of Acine‐tobacter baumannii is increasing in recent year in our hospital ,as well the resistance rate to the common Antibiotics .Monitoring the resistance of Acinetobacter baumannii should be strengthened for preventing resistant bacteria from spreading .

Objective To investigate the characteristics of HBV replication in HepG2.2. 15 cells treated with LAM alone or sequentially treated with LAM and IFN-α, and to further explore the different suppressive effect on HBV replication by LAM and IFN-α in vitro. Methods Untreated HepG2. 2. 15 cells were used as control group, HepG2. 2. 15 cells treated with 1 000 IU/ml of IFN-α alone for 10 d were served as IFN-α group. The HepG2.2. 15 cells treated with 0.2,1,5,20,100 μmol/L of LAM were used as LAM groups. HepG2. 2. 15 cells treated with 0. 04,0. 2,5,25,125,200 μmol/L of LAM for 7 d, then combined with 1 000 IU/ml of IFN-α for another 3 d. Afterwards, the cells were treated with 1 000 IU/ml of IFN-α only for another 10 d. These cells were served as LAM/IFN-α sequential treatment group. The ELISA was used for analyzing the secreted HBV antigens, while the Dot blot, Southern blot were applied for analyzing the extracellular HBV DNA and intracellular HBV replicative intermediate DNA in HepG2. 2. 15 cells of different treatment groups. Results The secreted HBsAg in the LAM group were 1. 77 ± 0. 22, 1.65 ±0.25, 1.95 ±0. 19, 1.34 ±0. 11, 1.07 ±0.05, respectively, and the secretion of HBeAg were 1.41 ±0. 13, 1.37 ± 0. 09, 1.63 ± 0. 07, 1.26 ± 0. 12, 1.05 ± 0. 09. The secreted HBsAg and HBeAg in control group were 3. 34 ± 0. 15 and 3.33 ± 0. 05. Statistical analysis showed that HBsAg and HBeAg secretion in the LAM group were significantly reduced by treatment with LAM. The t values of HBsAg were 10. 21,10.04, 9.94, 18.62, 24.86, and the t values of HBeAg were 23.87, 32.97, 34.22, 27.57, 38.35,respectively, all P values were ＜ 0.05. Dot blot, Northern blot hybridization analysis indicated that the extracellular HBV DNA and intracellular HBV replicative intermediate DNA could not be detected in the LAM group after cells treated by LAM for 10 days. When LAM was replaced with treatment of 1 000 IU/ml of IFN-α alone, it could not suppress the HBV replication effectively. Moreover, the intracellular HBV replicative intermediate DNA still existed in almost all groups, accompanied with the recovered expression of HBV antigens as well as extracellular HBV DNA, which suggested that the HBV particles restored replication again and secreted extracellular in HepG2. 2. 15 cells, although the sequential treatment lasted for 10 days.Conclusion The effect of viral suppression by LAM and IFN-α in vitro were different, which attributed to the different HBV replicative characteristcs in HepG2. 2. 15 cells.

Objective To observe the expression levels of Mig in the patients with chronic hepatitis B .Methods The study pop‐ulation consisted of 88 chronic hepatitis B patients and 53 healthy controls .The ELISA ,RT‐PCR and Western‐blotting were used for analysing the expression levels of Mig in serum ,peripheral blood mononuclear cells and liver tissue of the patients with chronic hepatitis B ,while the immunohistochemistry was applied for analysing the distribution of Mig in liver tissue .Results The expres‐sion of Mig in serum ,peripheral blood mononuclear cells and liver tissue of the chronic hepatitis B patients with HBeAg negative were (247 .03 ± 63 .14)pg/mL ,(0 .95 ± 0 .21) ,(0 .79 ± 0 .23) ,and that in the chronic hepatitis B patients with HBeAg positive were (243 .05 ± 53 .00)pg/mL ,(0 .98 ± 0 .35) ,(0 .74 ± 0 .18) ,which were both significantly higher than those in healthy controls ,the difference was statistically significant(P<0 .05) .Conclusion Increased levels of Mig in the patients with chronic hepatitis B may be related to immune state of patients .

Objective To study the relationship between serum Treg cells content and superoxide dismutase(SOD) level with the severity and clinical prognosis in the patients with preeclampsia.Methods Forty cases of preeclampsia in our hospital from September 2015 to September 2016 were selected as the research subjects divided into the mild preeclampsia group(25 cases) and severe preeclampsia groups(15 cases) according to the disease severity,and 20 healthy pregnant women served as the control group.The peripheral blood Treg cells and serum SOD level were compared between the two groups.The relationship between Treg cell content and serum SOD level with the disease severity and prognosis in the patients with preeclampsia was investigated.Results The systolic and diastolic blood pressure in the severe preeclampsia group were(145.3±10.6)mm Hg and(102.3±7.8)mm Hg,the occurrence rates of premature delivery,placental abruption and fetal distress were 33.3%,20.0% and 13.3% respectively,BMI before pregnancy was 25.4±1.3,which were significantly higher than those in the mild preeclampsia group and control group,while the mild preeclampsia group was higher than the control group.The Treg cell content and SOD level in the severe preeclampsia group were (2.3±0.7)×106/L and(2.6±0.7)μg/mL,which were significantly lower than those in the mild preeclampsia group and control group and which in the mild preeclampsia group were (3.4±0.6)×106/L and(4.3±0.9)μg/mL,and significantly lower than those in the control group,the differences were statistically significant(P<0.05).The Logistic regression analysis results indicated that BMI index(P=0.018),Treg cell content(P=0.024) and SOD level(P=0.029) were the independent risk factors for poor pregnancy outcome.Conclusion The peripheral blood Treg cells content and serum SOD level are closely related to poor pregnancy outcome in the patients with preeclampsia.

Aim To construct and express the eukary-otic expression vector of double-stranded RNA-depend-ent protein kinase (PKR)fusion green fluorescent and analyse its antiviral activity of HBV in vitro.Methods The PKR gene was cloned into an empty expression vector pEGFP-N1 using molecular clone technology. After being confirmed by restriction enzyme digestion and sequencing methods,the recombinant plasmid was named as pEGFP-PKR that was subsequently transfect-ed into HepG2.2.15 cells using LipofectamineTM2000. The expression level of PKR in HepG2.2.15 cells was confirmed by using fluorescent microscopy. Mean-while,HBV DNA and HBsAg/HBeAg were detected by real-time PCR and electrochemiluminescence meth-od,respectively.Results Both restriction enyme di-gestion and sequencing assays showed that the recombi-nant vector pEGFP-PKR was successfully constructed in our study.Fluorescent microscopy observation indi-cated that the fusion protein pEGFP-PKR expressed ef-ficiently in HepG2.2.15 cells.Moreover,compared with the empty vector group,the expression of HBV antigen in supernatants was significantly decreased (P<0.05 ).However,the extracellular HBV DNA ex-pression was not inhibited significantly.Conclusion In vitro,PKR proteion has certain antiviral activity of HBV.

Objective To evaluation the method of rapid detection of Methicillin-resistant Stphylococcus aureus (MRSA) ST239 clones with multiplex PCR assay and investigation of the epidemic status of MRSA blood stream infections in Hefei area.Methods Antibiotic susceptibility testing were applied to MRSA isolates from bloodstream infection,rapid screening and confirmation of MRSA ST239 clones by using multiplex PCR,Multilocus Sequence typing (MLST) and Staphyloccoccal Cassette Chromosome mec(SCCmec) typing.Results 51 of 106 clinic isolates Staphylococcus aureus were identified as MRSA,accounting for 48.1％.The resistance rate of MRSA to erythromycin,aminoglycosides and quinolone were significantly higher than Methicillin Sensitive Staphylococcus aureus (MSSA).Both MRSA and MSSA had a high sensitivity to cotrimoxazole,the sensivity rates were 86.3％ and 94.5％,respectively; 47 of 51 isolates of MRSA that detected by MRSA ST239 rapid screening were ST239 clones.Randomly selected 20 positive screening stains were confirmed as MRSA-ST239-SCCmec Ⅲ by MLST and SCCmectyping.Conclusions In Hefei area,nearly half of MRSA bloodstream infections in clinical isolates are MRSA-ST239-SCCmeclⅢ type and serious multidrug-resistance.The rapid detection of ST239 clones by multiplex PCR is a reliable and effective method for large-scale screening in laboratory.

OBJECTIVETo compare the CC-chemokine ligand 18 (CCL18) expression in the serum and malignant pleural effusion (MPE) of NSCLC patients and explore its regulatory effect on differentiation of monocyte-derived dendritic cells (Mo-DC).

METHODSCCL18 levels in the serum and MPE from 62 NSCLC patients were quantitated by immunoassay. CCL18 in sera from 26 healthy individuals, 28 exudative pleural effusions from inflammatory pulmonary diseases and 17 transudative pleural effusions from non-inflammatory diseases were used as control. Mo-DC was generated by culturing NSCLC-derived monocytes with GM-CSF and IL-4 in the presence or absence of CCL18. The mean fluorescent intensity (MFI) of CD14, CD80, CD83, CD86 and HLA-DR were analyzed by flow cytometry (FCM). Mo-DC was then co-cultured with purified T cells and the percence of CD25(+)FoxP3(+) cells was assayed by FCM.

RESULTSCCL18 levels in the sera of NSCLC patients and healthy individuals were (132.70 ± 15.52) ng/ml and (18.44 ± 0.99) ng/ml, respectively (P < 0.001). The levels of CCL18 in MPE, exudative PE and transudative PE were (155.6 ± 13.58) ng/ml, (190.4 ± 22.33) ng/ml and (20.89 ± 3.03) ng/ml, respectively. CCL18 in the MPE was significantly higher than that in transudates (P < 0.001), however, no significant difference was observed between CCL18 expression in exudative PE and MPE (P = 0.172). Of note, a moderate positive correlation (r = 0.421, P < 0.01) was observed between CCL18 levels in the paired MPE and serum of NSCLC. In the healthy control group, Mo-DC cultured in the presence of CCL18 showed 31.4 ± 15.8 (MFI) of CD14 expression, which was significantly higher than that in Mo-DC cultured in the absence of CCL18 (18.5 ± 8.9, P < 0.05). In contrast, the expressions of MFI of CD80, CD83, CD86 and HLA-DR were significantly decreased upon CCL18 induction (P < 0.05). In the NSCLC group, GM-CSF+IL-4+CCL18 induced a MFI of 45.2 ± 13.8 of CD14 expression in Mo-DC, which was also significantly higher than that of GM-CSF+ IL-4 induction (22.6 ± 10.5, P < 0.01). Similarly, the expressions of MFI of CD80, CD83, CD86 and HLA-DR were significantly decreased in the presence of CCL18 (P < 0.05). Furthermore, the MFI of CD14, CD83, CD86 and HLA-DR had significant differences between GM-CSF/IL-4/CCL18-induced Mo-DC derived from NSCLC patients and healthy control (P < 0.05). Finally, CD4(+) T cells co-cultured with NSCLC-derived, GM-CSF/IL-4/CCL18-treated Mo-DC had significantly higher percent of CD25(+)FoxP3(+) cells compared with that of CD4(+) T cells stimulated with Mo-DC induced by GM-CSF/IL-4(P < 0.01).

CONCLUSIONSCCL18 is present at a high level in MPE and serum of NSCLC patients complicated with pleural effusion and a moderate positive correlation exists between CCL18 levels in the two fluids. CCL18 inhibits maturation of Mo-DC, which consequently stimulates T cells to differentiate into CD25(+)FoxP3(+) regulatory T cells.

Carcinoma, Non-Small-Cell Lung ; metabolism ; Cell Differentiation ; Chemokines ; Chemokines, CC ; metabolism ; Coculture Techniques ; Dendritic Cells ; metabolism ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; metabolism ; Humans ; Interleukin-4 ; metabolism ; Ligands ; Lung Neoplasms ; Monocytes ; physiology ; Pleural Effusion ; T-Lymphocytes, Regulatory