Objective To study the alteration of surfactant protein A and D(sp-A,SP-D)result-ing from pneumcystis carinil pneumonia(PCP)and investigate its implication in the pathogenesis of PCP.Methods SD rat models of PCP were induced by subcutaneous injection of 25 mg cortisone acetate,normal control and negative control as well as bacterial pneumonia group were set up for comparison.During 8～12weeks.broncboalveolar lavage fluid (BALF) of rats was collected.Total nucleate cells of BALF were counted and differentiated as well as the concentrations of surfactant protein A(SP-A)and surfactant pro-tein D(SP-D)were measured by immunoblotting assay.Results The rats were divided into three im-munosuppressive groups,plus a norrflal control group. Group A: normal control(n=6)consisted of healthy SD rats;group B:negative control(n=6)employed rats with cortisone acetate injection over 8weekz without tung infection;group C:bacterial pneumonia(n=11),rats were injected with cortisone ac-etate over 8 weeks and resulted in bacterial pneumonia without other pathogens isolated;group D(n=14):rats were injected with cortisone acetate during 8～12 weeks and resulted in PCP without other pathogens isolated.During PCP infection,the total cell counts and the percentage of polymorphonuclears (PMNs)in BALF were significantly increased(P<0.01),but were lower than those in the bacterial pneumonia group.The concentration of SP-A of BALF in PCP(45.1 μg/ml 4±22.1 μg/m1)was signifi-cantly increased in comparison with that in negative control group(16.2 μg/ml±9.9 gg/ml,P<0.05)and that in bacterial pneumonia group(6.2 μg/ml±5.6 μg/ml,P<0.001).We also found that the rela-tive content of SP-D was significantly higher in PCP(24 249±4 780 grey values)than that in both nega-tive control(13 384±2 887 grey values,P<0.001)and bacterial pneumonia group(11 989±2 750 grey values,P<0.001).SP-A and SP-D were also higher in moderate to severe group of PCP than those seen in mild group(P<0.01,P<0.001).Conclusion There was obvious increase of SP-A and SP-D in PCP rats,and particularly,the change of which was greater than that in bacterial pneumonia.Therefore,the alteration of SP-A and SP-D may be of implication in the prevention and management of PCP.

Objective To establish a cell model that is close to the HCV replication in vivo and can support long-term HCV replication in vitro.Methods A human liver cell line 7721 was inoculated with serum from a chronic hepatitis C patient for 8 hours.After incubation,the presence of HCV RNA,the expression of HCV antigen and the location of HCV RNA in cells and/or supernatant were detected by RTPCR,immunohistochemistry and in situ hybridization respectively.Results It was found that plus-and minus-strand of HCV RNA could be discontinuously detected in both cells and supernatant as late as 66 days after inoculation even if cells had been subcultured for 6 times.HCV NS3、NS5 antigens could be expressed in cells and HCV RNA was mainly located within cytoplasm.Conclusion The above results suggested that 7721 cell line was not only susceptible to HCV but also could support its long-term replication in vitro.This HCV infection model in vitro was proved to be a useful tool for studying the mechanism of HCV infection and replication,as well as evaluating the antiviral compounds and screening the protective antibodies/vaccines.

Objective To analyze the epidemiological characteristics, clinical manifestations, diagnosis and treatment effect of brucella in=nfection, and to provide practical foundation and valuable experience for clinical treatment and prevention of brucellosis.Methods One hundred and nineteen patients with brucella infection hospitalized at the First Hospital of Lanzhou University from January 1st 2011 to July 30th 2011 were enrolled.Clinical data including gender, age, contact history with bovine and clinical manifestations such as fever, sweating and joint and muscle pain were extracted.Diagnosis and treatment effect of these patients were all recorded.The epidemiological characteristics and clinical manifestations were analyzed.Results The majority of patients were middle-aged male, and the number of male cases were 2.5 times of female cases.The brucella infections were observed all seasons in Lanzhou, while most cases were observed between May and July.A total of 78 cases had clear animal contact history.Among them, 53 cases (44.54%) had contact history with bovine.Main clinical manifestations included fever (n=84, 70.59%), muscle and joint pain (n=93, 78.15%), sweating (n=52, 43.70%), fatigue (n=29, 24.37%), and the most common clinical symptoms was joint and muscle pain.Blood routine test showed neutropenia (n=25, 21.01%), increase in blood sedimentation (n=34, 28.57%) and blood coagulation dysfunction (n=30, 25.21%).Liver function examination showed increase in ALT (n=37, 31.09%) and AST (n=43, 36.13%).Other examination showed that splenomegaly (n=28, 23.52%) was the most common finding, and liver diffuse lesions (n=22, 18.49%) ranked second.The most common drugs were doxycycline (n=79, 66.39%), rifampin (n=60, 50.42%) and cephalosporins (n=50, 42.02%).Among them, 45 cases (37.82%) were treated with combination therapy of rifampin and doxycycline, and 31 cases were treated with combination therapy of doxycycline and rifapentine or cephalosporins.Totally 112 cases (94.12%) recovered.Conclusions The majority of patients with brucellosis are male.The clinical manifestations of brucellosis are varied.Clinicians should pay attention to identify and prevent misdiagnosis.The timely and effective treatment should be provided according to patients' conditions.The relevant departments should also strengthen supervision and education of brucellosis.

Objective To investigate the antibiotic resistance and resistance genes of carbapenem-resistant Enterobacteriaceaes (CRE) isolated from 5 hospitals in Northeast China.Methods This study collected 85 CRE isolates during January 2013 to June 2015 from five hospitals in Northeast China.Drug sensitivities of 14 antimicrobial agents were determined by the broth microdilution method.The phenotypes of carbapenemases were screened by modified Hodge test and EDTA test respectively.The genotypes of carbapenemases and other extended spectrum β-lactamases (ESBL) were detected by PCR gene amplification and DNA sequencing method.Using the PCR result as gold standard, the performances of other two carbapenemase detection methods were evaluated.Results Among the 85 CRE strains collected in this study, Klebsiella pneumoniae was the most frequently isolated species (61/85,71.8%).The results of antimicrobial agent sensitivity showed that the 85 CRE strains had resistance rate of cephalosporin and β-lactams/enzyme inhibitor (piperacillin-tazobactam) over 80.0%.The resistance rate of carbapenem was high, with ertapenem 100.0% (85/85), meropenem 65.9% (56/85), imipenem 71.8% (61/85).There were 36 isolates resistant to both meropenem and imipenem.For fluoroquinolones, the resistance rates of levofloxacin and ciprofloxacin were 72.9% (62/85) and 65.9% (56/85), respectively.The resistance rate to fosfomycin and amikacin were 65.0% (55/85) and 54.1% (46/85), respectively.The resistance rate of colistin (21.2%, 18/85) and tigecycline (20.5%, 17/85) were low.Forty-nine strains were modified Hodge test positive and 12 strains were EDTA test positive.By PCR gene amplification and DNA sequencing method, 64 strains carried carbapenemase-encoding genes, of which KPC-2 was the main type (53/85, 62.4%), followed by IMP-4 (10/85, 11.8%), NDM-5 (7/85, 8.2%) and NDM-6 (1/85, 1.2%).At the same time, 85 CRE isolates had the ESBL gene detection and 47 isolates were CTX-M type ESBLs (47/85, 55.3%), with no TEM or SHV type.Conclusions Klebsiella pneumoniae is the majority of CRE strains from 5 large hospitals in Northeastern China.The CRE strains are resistant to most of antimicrobials.Most carbapenemases-producing isolates have the KPC-2 type.Nearly half of the carbapenemase-producing strains also carry ESBL genes, which makes the resistance mechanisms more complicated.

Objective To compare the clinical characteristics and prognosis of bacterial liver abscess in patients with or without type 2 diabetes mellitus, in order to guide clinical diagnosis and treatment.Methods The clinical data of 110 patients with bacterial liver abscess at the First Affiliated Hospital of Anhui Medical University were retrospectively analyzed.Among them, 45 cases with type 2 diabetes mellitus and 65 cases without type 2 diabetes mellitus were included in the study.The demographic data, clinical signs and symptoms, laboratory findings, pathogenic results, imaging findings, treatment and prognosis were compared between the two groups.Differences among the quantitative data with normal distribution were compared using t test, while count data were compared with χ2 test or Fisher exact test.Multivariate Logistic regression was used to determine the prognostic risk factors of two groups.Results Fever as initial symptom in diabetic group and non-diabetic group were 37 cases and 40 cases, respectively, while abdominal pain presented in 7 cases and 22 cases of the two groups, respectively, both with statistically significant difference (χ2=5.417 and 4.582, respectively, both P<0.05).As for laboratory examination, neutrophil counts in the two groups were (12.87±8.83)×109/L and (10.24±4.86)×109/L, respectively, the percentages of neutrophils were 0.841±0.077 and 0.799±0.103, respectively, albumin levels were (28.36±4.65) g/L and (30.67±6.16) g/L, respectively, with statistically significant difference (t=2.010, 2.317 and-2.265, respectively, all P<0.05).Patients with elevated blood urea nitrogen in the two groups were 13 cases and 8 cases, respectively, patients with elevated creatinine were 9 cases and 4 cases, respectively, with statistically significant difference (χ2=4.733 and 4.892, respectively, both P<0.05).In diabetic group, pus culture was positive in 13 out of 19 cases, and blood culture was positive in 7 out of 21 cases.In non-diabetic group, pus culture was positive in 9 out of 13 cases, and blood culture was positive in 6 out of 25 cases.The positive rates of Klebsiella pneumoniae in the two groups were 37.5% (15/40) and 15.8% (6/38) , respectively, with statistically significant difference (χ2=4.669, P=0.031).The effective rates of the two groups with glycemic control were 84.4% (38/45) and 84.6% (55/65), respectively, and the mortality rates of the two groups were 2.2% (1/45) and 1.5% (1/65), respectively.Multivariate Logistic regression showed that multiple abscess (OR=34.61, 95% CI: 1.601-748.457, P=0.024) was prognostic risk factor and invasive intervention (OR=0.028, 95%CI: 0.001-0.984, P=0.049) was protective factor in the diabetes mellitus group.Hypoalbuminemia (OR=14.793, 95% CI: 1.605-136.322, P=0.017) and the history of abdominal surgery within two years (OR=7.624, 95% CI: 1.294-44.913, P=0.025) were prognostic risk factors in the non-diabetic group.Conclusions Patients with bacterial liver abscess showing symptoms of severe infection in diabetic group are more frequently than patients in non-diabetic group.Klebsiella pneumoniae infection rate is also higher in diabetic group.Patients with bacterial liver abscess and diabetes mellitus should receive antibiotic treatment combined with invasive therapy in time.

Objective To explore the distribution of parvovirus B19 (HPVB19) infection in patients with leukopenia.Methods Patients who visited the Affiliated Hospital of Hangzhou Normal University from January 2015 to June 2016 were analyzed.Patients with peripheral leucocytes count less than 3.5×109/L were included in experiment group and healthy people were included in control group.HPVB19 IgG and IgM were detected by enzyme-linked immunosorbent assay, and HPVB19 DNA was detected by quantitative polymerase chain reaction.Differences in continuous data between two groups were compared with two-sample t test and those in categorical data were compared with Chi-square test.Results A total of 79 patients were included in experiment group, including 32 males and 47 females.Ages ranged from 24 to 62 years old.And 126 healthy individuals were included in control group, including 55 males and 71 females.Ages ranged from 28 to 67 years old.The positive rates of HPVB19 IgG, IgM and DNA in experiment group were 34.2%, 5.1% and 3.8%, respectively, while those in control group were 36.5%, 0 and 0, respectively.The detection rates of HPVB19 IgM and DNA between two groups were significantly different (χ2=6.507, P=0.011 and χ2=4.856, P=0.028, respectively).Sequence analysis for 3 of the HPVB19 DNA positive samples showed that there were two single nucleotide polymorphisms in VP1/VP2 sequence from one patient, which contributed to the 153rd (L/H) and 219th (N/Y) amino acids mutations, respectively.Phylogenetic analysis found that two strains belong to genotype 1a and one strain belongs to genotype 1b.Conclusions Detection rate of parvovirus HPVB19 infection (positive rates of HPVB19 IgM and DNA) in leukopenic patients is significantly higher than healthy controls.HPVB19 should be detected before considering transfusion in leukopenic patients in clinical practice.

Objective To investigate the relationship between interleukin (IL)-28B gene polymorphisms (rs12979860 and rs8099917) and treatment response in patients with chronic hepatitis C in China.Methods Taqman probes single nucleotide polymorphism genotyping methods were used to detect the genotypes of rs12979860 (C/T) and rs8099917 (T/G) located at IL-28B gene in 105 included patients.The patients were treated with standard doses of pegylated interferon plus ribavirin and were followed up regularly for therapeutic response and adverse reaction.The relationship between IL-28B gene polymorphism and antiviral treatment response of patients were analyzed.Categorical data were analyzed using Pearson chi-square test or Fisher exact test.Results Totally 105 cases were included in our study and 2 cases lost to follow-up because of moving away.Eight-one cases (78.6%) of the remaining 103 patients were CC/TT genotype (CC/TT group) at rs12979860 and rs8099917, 19 cases (18.4%) were CT/TG (CT/TG group) and 3 cases (2.9%)were TT/TG (TT/TG group).No other genotypes were detected and linkage disequilibrium was discovered at the two polymorphism loci (r2=0.11).After 4 weeks of treatment, 35 cases (43.2%) in CC/TT group, 3 cases (15.8%) in CT/TG group and non in TT/TG group achieved rapid virological response (RVR).There were statistically significant differences among three groups (P=0.033).After 12 weeks of treatment, 45 cases (55.6%) in CC/TT group, 6 cases (31.6%) in CT/TG group and none in TT/TG group achieved early virological response (EVR).There were statistically significant differences among three groups (P=0.025).At the end of the treatment, 68 cases (83.9%) in CC/TT group, 10 cases (52.6%) in CT/TG group and only 1 case (33.3%) in TT/TG group achieved end-of-treatment response (ETR).There were significant statistical differences among the three groups (P=0.003).After 24 weeks of follow-up, 62 cases (76.5%) in CC/TT group, 9 cases (47.4%) in CT/TG group and 1 case (33.3%) in TT/TG group achieved sustained virological response (SVR).There were statistically significant differences among the three groups (P=0.014).One hundred and one cases in CC/TT group developed adverse events, among them 19 cases needed clinical treatment.There were 43 cases in CT/TG group developed adverse events and 9 cases needed treatment.Seven cases in TT/TG group developed adverse events and only 1 case needed treatment.There were no statistically significant difference among three groups (χ2=0.139,P>0.05).Conclusions The genotype of rs12979860 (C/T) and rs8099917 (T/G) at IL-28B gene could affect the treatment response in patients with chronic hepatitis C.RVR and SVR are higher in patients with genotype CC/TT, which might help to guide HCV treatment.

Objective To analyze the electroencephalograph (EEG) features of 43 patients with clinically possible or probable Creutzfeldt-Jakob disease (CJD) and its correlation with megnetic resonance imaging (MRI) imaging and clinical manifestations.Methods All patients diagnosed with suspected CJD who were hospitalized in Xuanwu Hospital from January 2013 to December 2015 were collected.The clinical data, EEG and MRI imaging features were analyzed retrospectively.Based on the periodic sharp wave complexes (PSWC) appearance in EEG results, the patients were divided into typically changed group (TCG), atypically changed group (ACG) and unchanged group (UCG).Age, disease duration, clinical manifestations and MRI features among three groups were analyzed and the correlations between patient′s EEG features and age, disease duration, clinical manifestations or MRI features were explored using spearman method.Results Among the 43 patients with possible or probable CJD disease, 26 were male and 17 were female with an average disease duration of 4 months.The age of onset ranged from 31 to 80 with an average of (58.0±9.8) years old, and 86.0% of patients were 51 years old or above.Clinical characteristics of CJD patients according to occurrence rate were as follows: 35 cases (81.4%) with cognitive impairment, 29 cases (67.4%) with mental and behavior disorder, 28 cases (65.1%) with pyramidal tract damage, 24 cases (55.8%) with cerebellar symptoms, 23 cases (53.5%) with extrapyramidal symptoms,17 cases (39.5%) with myoclonic, 13 cases (30.2%) with dyssomnia, 13 cases (30.2%) with visual disorder and 2 cases (4.7%) with akinetic mutism.Regarding EEG features, 39.53% (17/43) of patients showed typical periodic sharp wave complexes (PSWC) (TCG group), 51.2% (22/43) had irregular rhythm and different forms of slow wave (ACG group) and only 9.3% (4/43) had no EEG change (UCG group).The occurrence rate of ribbon sign in MRI was 82.4% (14/17) in TCG group, 77.3% (17/22) in ACG group and none in UCG group.The rates were significantly higher in TCG and ACG group than that in UCG group (both P<0.05).Spearman correlation analysis revealed that EEG features was correlated with disease duration (r=0.351, P=0.021) and visual impairment (r=-0.377, P=0.013) for all CJD patients.There was no correlation between EEG and MRI or other clinical manifestations such as myoclonic, age and so on (all P>0.05).Conclusions EEG showed typical changes associated with disease duration in different stages of disease.EEG and MRI are two different means to evaluate different aspects of patients with CJD disease, and combination of two means could achieve better evaluation results.

Objective To investigate the clinical indicators which can predict esophageal varices in patients with primary biliary cirrhosis (PBC).Methods A total of 351 patients with PBC from the First Affiliated Hospital of Xinjiang Medical University from January 2013 to December 2016 were retrospectively analyzed, including 173 patients with esophageal varices and 178 patients without varicose veins.The alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), γ-glutamyl transpeptadase (γ-GT), total bilirubin (TBil), albumin (Alb), prothrombin time (PT), platelet (PLT), AST to ALT ratio (AAR), fibrosis index based on the 4 fator (FIB-4), AST to PLT ratio index (APRI) and Mayo scores were compared between two groups.Group t test or rank sum test was used to compare the two groups.Relation between the indicators mentioned above and esophageal varices were tested by univariate analysis.Multivariate unconditional Logistic regression was used to screen these indicators to independently predict esophageal varices in PBC patients.Results Age, PT, TBil, AAR, FIB-4, APRI and Mayo scores of PBC patients with esophageal varices were all higher than those of patients without esophageal varices ([60.3±10.6] years old vs [51.9±10.9] years old, [13.31±3.12] s vs [11.17±2.42] s, 28.06 [18.05, 60.06] mmol/L vs 15.39 [10.64, 33.63] μmol/L, 1.69±0.91 vs 1.23±0.95, 6.18 [4.05,9.16] vs 1.80 [1.10,2.74], 1.95 [1.12,3.08] vs 0.69 [0.38,1.57], 6.45±1.52 vs4.62±1.53, respectively).Whereas ALT, γ-GT, Alb and PLT levels were all lower than those without varicose veins (36.60 [19.88, 74.28] U/L vs 59.32 [23.58, 132.70] U/L, 71.00 [38.36, 165.38] U/L vs 125.00 [37.50, 336.21] U/L, [29.78±6.33] g/L vs [39.51±25.16] g/L, [103.43±52.84]×109/L vs [234.44±90.40]×109/L, respectively).The differences were all statistically significant (t=-7.25, t=-7.18, Z=-5.823, t=-4.60, Z=-8.427, Z=-12.661, t=-11.25, Z=-3.218, Z=2.987, t=4.94, t=16.63, respectively;all P<0.01).Multivariate unconditional logistic regression analysis showed that PLT<149×109/L (OR=0.966, 95% CI: 0.957-0.974), PT>11.95 s (OR=0.705, 95%CI: 0.569-0.874), TBil>17.19 μmol/L (OR=0.99, 95%CI: 0.982-0.999), FIB-4>3.02 (OR=0.868, 95% CI: 0.807-0.932) and Mayo score>4.88 (OR=6.053, 95%CI: 2.388-15.342) were independent clinical indicators for the prediction of esophageal varicose veins.Conclusions PLT, PT, TBil, FIB-4, and Mayo scores can be used as predictors of esophageal varices in patients with PBC.

Objective To study the epidemiology and genotypes of extended-spectrum beta-lactamases (ESBL)-producing Escherichia coli (EC) and Klebsiella pneumoniae (KP) that caused community-onset bloodstream infections (COBSI) in 9 county hospitals of Zhejiang Province.Methods This is a multi-center, prospective, observational study.The cases and isolates with COBSI caused by EC and KP were consecutively collected from 9 county hospitals in Zhejiang Province between 1st March 2014 and 30th April 2015.The double disk diffusion method was used to confirm the production of ESBL.The ESBL genotypes were determined by polymerase chain reaction(PCR) amplification and sequence analysis.Multi-locus Sequence Typing (MLST) was used to analyze the homology of ESBL-producing isolates.Minimal inhibitory concentration (MIC) of frequently used drugs for ESBL-producing isolates was determined by in-vitro antimicrobial susceptibility tests.Results During the study period, a total of 172 cases with COBSI were collected and 171 cases were eligible, among which 126 were caused by EC and 45 were caused by KP.The overall prevalence of ESBL was 34.5% (59/171),and the prevalence of ESBL-EC and-KP was 41.3% (52/126) and 15.6% (7/45), respectively.CTX-M-type ESBL accounted for 96.6% (57/59) of all the ESBLs-producing isolates, and the most common type was CTX-M-14 (27.1%, 16/59), followed by CTX-M-55 (22%, 13/59).MLST analyses revealed significant genetic diversity among ESBL-EC and-KP.The most prevalent ST of ESBL-EC was ST131 (23.1%).In addition to carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam, moxalactam, amikacin and fosfomycin also showed good in-vitro activity against ESBL-EC and-KP.Conclusions The prevalence of ESBL in EC and KP is high in 9 county hospitals of Zhejiang Province, and the most common genotypes are CTX-M-14 and CTX-M-55.The detection rate of ESBL in EC is higher than in KP.It could be considered adequate empirical therapy according to the results of antimicrobial susceptibility tests.Carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam, moxalactam, amikacin and fosfomycin have good in-vitro activity against ESBL-EC and-KP.