Objective To construct the mice pneumonia model with imipenem-resistant Acinetobacter baumannii and provide experimental model in anti-pan-resistant Acinetobacter baumannii therapy study. Methods A total number of 120 4-week-old BALB/C male mice were randomly selected and divided into three groups including micro-intratracheal injection, ultrasonic atomizing and nasal dripping. The mice were treated with methotrexate to induce hypo-immunity in every group. These BALB/C mice of normal immunity and hypo-immunity were infected through Imipenem-resistant Acinetobacter baumannii by microintratracheal injection, ultrasonic atomizing and nasal dripping, respectively. The morbidity, mortality,bacterial clearance rate and histopathology in lung were determined. Results The morbidities of BALB/C mice with hypo-immunity infected by micro-intratracheal injection and ultrasonic atomizing achieved 100%(30/30), while the mortalities were 100% (10/10) and 33.3% (3/10), respectively. Mice in two groups above displayed the influx of neutrophils, lymphocytes and macrophages in the peri-bronchial and alveolar interstitial space 12-24 h after pulmonary infection. In addtion, the mice in micro-intratracheal injection group displayed coUapse of partial alveolar walls, formation of abscesses and bacterial colonies in alveoli. While the lung pathology in mice of ultrasonic atomizing group was characterized by cell degeneration in some regions in the lungs, slight relaxation, congestion in alveolar wall vessels and normal of bronchial and alveolar tissue 24 h after inoculation. Degeneration in peri-tracheal and peri-bronchial areas was observed 24-48 h after inoculation, along with highly expanded pulmonary blood vessels and edems. The inflammation was reduced at 48 hours. There was no obvious pulmonary infection in BALB/C mice with hypo-immunity by nasal dripping with mortality of 0% (0/10) and no significant histopathologic change in lungs. Conclusions BALB/C mice with hypo-immunity pneumonia model with Imipenem-resistant Acinetobacter baumannii can be conducted by micro-intratracheal injection or ultrasonic atomizing, but the latter has the advantages of high-productivity, easy-operation, low-cost, time-saving and usefulness. Mice with normal immunity are not susceptible to imipenem-resistant Acinetobacter baumannii.

Objective To evaluate the differences of α-fetoprotein (AFP), ffeeβ-human chorionic gnnadotropin (HCG) indexes in 3 foreign median databases for antepartum risk screening, and establish the median databases of normal pregnant women in Wenzhou for antepartum screening of AFP, free-β-HCG indexes through the suitable median computational models. Methods The levels of AFP and free β-HCG of 20054 normal pregnant women in Wenzhou were detected by time-resolved fluorometry. The data in this paper were compared with the data of 2T-risk ( 2T), Lifecycle-2. 2 (LC2.2 ) and Lifecycle -3.0 (LC3.0) by double-factor ANOVA. The differences between the data in the paper and the data from Shenyang and Ningbo were analyzed. The median database of Wenzhou pregnant women was established by the suitable regression model, with the stability of nonlinear regression models of the 3 software assessed by model correcting fitting, distribution mean of model fitting logarithmic and standard deviation. Results The levels of AFP and freeβ- hCG reported here were 10% and 16% higher than the data of 2T-risk, 15% and 20% higher than that of LC 2. 2, 6% and 17% higher than that of LC 3.0 respectively. The differences were statistically significant. ( FAFP = 161. 757 ,P < 0. 01 ; Ffree-β-HCG = 58. 261, P < 0.01 ). The levels of AFP and free β- hCG in Wenzhou were 2% higher and 3% lower than that of Shenyang, 1% and 2% higher than that of Ningbo. There was no statistical difference of AFP levels among Wenzhou , Shenyang and Ningbo( FAFP = 0. 174 ,P =0. 840) while the differences of free-β-hCG were statistically significant( F<,free-β-HCG> = 13.303 ,P < 0. 01 ). The differences of quadratic equation regression model, exponent quadratic function regression model and exponent quadru-function regression model of 2T, LC-2. 2 and LC-3.0 were not remarkable. The exponent quadru-function regression model was the best. Conclusions There are significant differences between the data from Wenzhou, Shenyang and Ningbo and the data of T-2 risk, LC-2. 2, LC -3.0. The discrepancy is due to the ethnic and different parameters of regression models. So the model parameters and the median databases are urgently required in China. The differences of large sample size of AFP from Wenzhow, Shengyang and Ningbo are not significant, while the differences of free-β- hCG from Wenzhow, Shengyang and Ningbo is remained because of its instability. The levels from Wenzhow and Ningbo are near. It is suggested that the laboratories with small sample size can establish their own laboratory parameters using the reference obtained from large sample size under the same experimental conditions. There are no significant differences of stability among regression computational models in the 3 software. The exponent quadru-function regression model can be used to establish the median databases for the screening with the similar data distribution in the paper.

Objective To investigate the correlation of T-lymphocyte expressing HLA-DR with serum HBV DNA and HBeAg contents in chronic hepatitis B. Methods Totally 134 chronic hepatitis B patients and 36 healthy blood donors were enrolled in the study. The T-lymphocytes (CD3 + HLA-DR + ,CD4 + HLA-DR+ and CD8 + HLA-DR+ T) expressing HLA-DR were detected by flow cytometry, the serum HBV viral loads were detected by the real-time quantitative PCR and HBeAg was detected by chemiluminescence method. According to serum HBV DNA viral loads patients were defined as HBV DNA negative (≤ 103 copies/mL), low (＞ 103 - 105 copies/mL), medium (＞ 105 - 107 copies/mL) and high groups (＞ 107 - 109 copies/mL) ; according to serum HBeAg levels, patients were defined as HBeAg negative (≤1 PEIU/mL), low (＞ 1 - 100 PEIU/mL), medium (＞ 100-1 000 PEIU/mL) and high groups (＞ 1 000-10 000 PEIU/mL). T test and one-way ANOVA were performed. Results With HBV DNA loads, HBeAg levels increased, the percentage of CD3 + HLA-DR + , CD4 + HLA-DR + and CD8 + HLA-DR +decreased, especially CD8 + HLA-DR +. Compared with HBV DNA negative group, the percentages of CD3 +HLA-DR + , CD4 + HLA-DR + and CD8 + HLA-DR + were significantly reduced in high group (t = 3. 686,4. 592 and 3. 216, P ＜ 0. 0l); the percentages of CD4 + HLA-DR + and CD8 + HLA-DR + were also reduced in medium group (t = 3. 761 and 2.862, P ＜ 0.01); while in low group, only the percentage of CD8 + HLA-DR + was reduced (t = 2.215, P ＜ 0.05). Compared with HBeAg negative group, the percentages of CD3 +HLA-DR+, CD4 + HLA-DR+ and CD8 + HLA-DR+ were significantly reduced in medium and high groups (thigher =3. 144, 2.222 and 4.035; tmiddle =3.311, 2.362 and 3.374, P ＜0.05), while in the low group,only the percentage of CD8+HLA-DR+ was reduced (t=2.029, P＜0. 05). Conclusion The combined measurement of HBV DNA, HBeAg and T-lymphocytes expressing HLA-DR in chronic hepatitis B patients may not only help to evaluate the immune status of patients, but also can predict the disease progression and clinical outcomes.

Objective To understand the genotypic characteristics and the neutralizing antibody levels of Japanese encephalitis virus(JEV)and Japanese encephalitis(JE)in both vector mosquitoes and in healthy people of Zhejiang province. Methods Virus was isolated from mosquitos sampled from the Monitoring Stations located in Xianju county during 2012 to 2013. Phylogenetic and homological studies were carried out on the E gene. A total of 1 263 blood specimens from 642 healthy people were collected before and after the seasons of JE epidemics. JEV neutralizing antibody was detected by the micro-neutralization test. Results Twenty-five JEV strains were isolated from a total of 11 650 mosquitoes. The identity of nucleotide appeared as 87.8%-99.7% both from 2012 to 2013 and from 1982 to 2010 while as 87.7%-88.0%with vaccine strain SA14-14-2,in Zhejiang. The phylogeny tree of E gene indicated that the newly isolated virus belonged to genotypeⅠ but no mutation of amino acid sequence coding conformational epitope was identified in the envelop protein. Both positive rates and the geometric mean titer(GMT)of neutralizing antibody in healthy people were 31.5%-42.0%and 1∶2.56-1∶3.53 in Xianju county,during 2012 and 2013,respectively. Both of the two positive rates(χ2≤1.76,P>0.05)and the two GMTs(u≤0.64,P>0.5)for antibodies pre or post the epidemic season did not show significant differences. Conclusion JEV isolated in Xianju during 2012 and 2013 belonged to genotypeⅠ. The positive rates of JEV neutralizing antibody from healthy people in Xianju were less than 42.0%,which showed no significant differendes pre or post JE epidemic season.

Objective To investigate the level of CD31 molecule expression on peripheral T lymphocytes of patients with chronic hepatitis B (CHB),and to study the relationship between the expression of CD31 molecule and serum ALT levels,serum levels of HBV DNA and HBeAg.Preliminarily explore the relationship between the expression frequency of CD31 molecule and patients CHB,in order to understand changes in different immune status when chronic HBV infecting.Methods A total of non-antiretroviral therapy 42 patients with CHB and 25 healthy volunteers were enrolled for the study.In peripheral blood,separated lymphocytes conventionally,the percentage of CD4 + 、CD8 + 、CD31 + 、CD31 + CD4 + 、CD31 + CD8 + 、CD31 + CD4 +/CD4 + 、CD31 + CD8 +/CD8 + cells were assessed by flow cytometry,serum ALT levels were tested by automatic biochemical analyzer,serum HBVDNA levels were determined by real-time fluorescent quantitative PCR,and the serological markers of HBV were examined through chemiluminescence simultaneously.Results The percentages of CD4 + 、CD31 + 、CD31 + CD4 + 、CD31 +CD4 +/CD4 + cells in patients with CHB were significantly less than the healthy(P ＜ 0.05);the percentages of CD31 + 、CD31 + CD4+ 、CD31 + CD8+ 、CD31 + CD4+/CD4+ cells in ALT ＜2ULN group of patients with CHB were significantly lower than ALT≥2ULN group(P ＜ 0.05);serum HBVDNA loads lower group (＞103-105copies/ml),middle group (＞ 105-107 copies/ml) and higher group of HBVDNA (＞ 107-109 copies/ml) CD31 molecule expression frequency were on the rise,but no statistically significant difference were observed in the comparison between groups (P ＞ 0.05);the percentages of CD31 + 、CD31 + CD8 + 、CD31 + CD4 +/CD4 + cells in the HBeAg-negative group were significantly lower than HBeAg-positive group (P ＜0.05).Conclusions The frequency of CD31 molecule expression on peripheral T lymphocytes of patients with CHB were declined;there is a certain relationship between the frequency of CD31 molecule expression and serum ALT levels,different loads of HBVDNA,HBeAg positive or negative in patients with CHB;chronic HBV infection of immune status may be associated with T cell surface expression of CD31 molecule frequency.

OBJECTIVETo understand the genotypic characteristics and the neutralizing antibody levels of Japanese encephalitis virus (JEV) and Japanese encephalitis (JE) in both vector mosquitoes and in healthy people of Zhejiang province.

METHODSVirus was isolated from mosquitos sampled from the Monitoring Stations located in Xianju county during 2012 to 2013. Phylogenetic and homological studies were carried out on the E gene. A total of 1 263 blood specimens from 642 healthy people were collected before and after the seasons of JE epidemics. JEV neutralizing antibody was detected by the micro-neutralization test.

RESULTSTwenty-five JEV strains were isolated from a total of 11 650 mosquitoes. The identity of nucleotide appeared as 87.8%-99.7% both from 2012 to 2013 and from 1982 to 2010 while as 87.7%-88.0% with vaccine strain SA14-14-2, in Zhejiang. The phylogeny tree of E gene indicated that the newly isolated virus belonged to genotype I but no mutation of amino acid sequence coding conformational epitope was identified in the envelop protein. Both positive rates and the geometric mean titer (GMT) of neutralizing antibody in healthy people were 31.5%-42.0% and 1 : 2.56-1 : 3.53 in Xianju county, during 2012 and 2013, respectively. Both of the two positive rates (χ(2)≤1.76, P > 0.05) and the two GMTs (u≤0.64, P > 0.5) for antibodies pre or post the epidemic season did not show significant differences.

CONCLUSIONJEV isolated in Xianju during 2012 and 2013 belonged to genotype I. The positive rates of JEV neutralizing antibody from healthy people in Xianju were less than 42.0%, which showed no significant differendes pre or post JE epidemic season.

Amino Acid Sequence ; Animals ; Antibodies, Neutralizing ; blood ; Antibodies, Viral ; blood ; China ; Culicidae ; virology ; Disease Vectors ; Encephalitis Virus, Japanese ; genetics ; immunology ; isolation & purification ; Encephalitis, Japanese ; virology ; Epitopes ; Genotype ; Humans ; Neutralization Tests ; Phylogeny