Objective To investigate the clinical characteristics, risk factors and prevention measures of carbapenem-resistant Klebsiella pneumoniae (CRKP) infection. Methods Thirty-nine cases whose CRKP isolation and culture were positive and who were confirmed with nosocomial infection in 2nd affiliated hospital medical school of Zhcjiang University between July 2006 and July 2008 were retrospectively investigated. Results There were 22 (56.4%) sputum specimens among CRKP samples. The average age of the patients was 64.0 years old and the average length of staying in hospital was 80.8 days. There wcre 36 patients who had been in intensive care unit (ICU) within 2 weeks and 26 were conscious disturbance,38 received urethral catheterization, 34 inserted with central venous catheter, 32 received tracheal intubation and 30 assisted with mechanical ventilation. There were 27 (69. 2 % ) patients who were co-infected with other bacteria and 8 were co-infccted with fungi. The mortality was 46. 2% in patients with CRKP infection. Before CRKP-positive culture, 27 (69.2%)experienced ＞4 kinds of antibiotics with the majority of β-lactams/β-lactamase inhibitors and carbapenems.Conclusions CRKP infection frequently occurrs in elder and patients who prolonged stay in hospital,especially patients in ICU. Invasive procedures could increase the infection opportunities.

Objective To screen immune molecules expressed on peripheral blood mononuclear cells (PBMCs) which arc related to the aggravation of chronc hepatitis B (CHB). Methods Real-time polymerase chain reaction (PCR) was employed to detect the relative expression of 39 immune molecules at mRNA level and then compare the differences between groups (control group, mild CHB group, moderate CHB group and severe CHB group). The investigated immune molecules included leukocyte differentiation antigens, chemokine receptors, apoptosis-related ligands and receptors,adhesion molecules and Toll like receptors. Scheffe model was used to compare the differences among all groups and Spearman rank correlation was used for correlation analysis. Results Among the 39 immune molecules, the expressions of tumor necrosis factor related apoptosis inducing ligand (TRAIL), TRAIL receptor (TRAIL-R) 2, CD64, CD30, CD27, CD28, L-selectin (CD62L),intercellular adhesion molecule-1 (ICAM-1), chemokine (C-X-C motif) receptor (CXCR) 2 were significantly increased in severe CHB group compared with those in control group ( 1.96, 2.13, 1.33,1.16, 1.57, 2.14, 2.03, 2. 10 and 2.09, respectively; all P＜0.05). The expressions of TRAIL,TRAIL-R2, CD64, CD30, CD27 were highly correlated with the expression of interferon gamma (IFN-γ) (r=0. 816, 0. 572, 0. 462, 0. 697 and 0. 793, respectively; all P＜0.01). The expressions of TRAIL-R2, CD64, CD30, CD62L, ICAM-1 were highly correlated with the expression of tumor necrosis factor alpha (TNF-α) (r=0. 494, 0. 588, 0. 568, 0. 968 and 0. 976, respectively; all P＜0.01). Conclusion The abnormal expression of TRAIL,TRAIL R2,CD64,CD30,CD27,CD28,CD62L,ICAM-1 and CXCR2 may play an important role in the pathogenesis and aggravatoin of hepatitis B.

Objective To explore the relationship between immune functions of patients with acute leukemia (AL) and invasive fungat infection (IFI). Methods T lymphocyte subpopulations and natural killer (NK) cells in 61 AL patients complicated with IFI at first visit, AL remission, the time of IFI onset and 4 weeks after antifungal treatment were measured by flow cytometry. Meanwhile,levels of IgG, IgM and IgA were detected by immunoturbidimetry. The statistical analysis was done using ANOVA, t test and chi-square test. Results CD3+ , CD3+ CD4+ , CD8+ CD28+ T lymphocyte as well as CD4+/CD8+ ratio at the time of IFI onset in AL patients were all lower than those at first visit, AL remission and 4 weeks after antifungal treatment (F= 25.6,26.6,13. 1,167.9; all P＜0.05), while CD8+ CD28- T lymphocyte were higher than those at first visit, AL remission and 4 weeks after antifungal treatment (F= 220.2,P＜0.01). CD3+ , CD3+ CD4+ and CD4 + /CD8+ ratio of patients who responded effectively to antifungal treatment were all higher than those of non-responders (t=3.75,8. 61,3.17; all P＜0.05). The serum levels of IgG, Igm and IgA at first visit, ALremission, the time of IFI onset and 4 weeks after treatment were similar (F=0.78,0.72,0.81; all P ＞0.05). The effect rate of antifungal therapy in AL remission group was higher than that in nonremission group (87% vs 53%,x2 = 7.62, P＜0.05). Conclusions The cellular immune functions are impaired severely in AL patients complicated with IFI, while the levels of IgG, IgM and IgA are similar during IFI. Therefore, the efficacy of antifungal therapy may partly depend on the recovery of cellular immune functions and remission of AL.

Objective To investigate whether the gene polymorphisms of cytochrome P450(CYP) 2E1 are associated with the risk of anti-tuberculosis drug induced hepatotoxity (ADIH).Methods In this case control study, 339 patients who matched the diagnosis criteria of tuberculosis were included. The gcneral healthy status and liver biochemical parameters were checked in all these patients. Polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) technique was used to determine CYP 2Et polymorphisms. The statistic analysis were performed by using both univariate and multivariate Logistic regression analysis. Results The allele frequencies of CYP 2E1 7632T/A, 1019C/T and 1259G/C in 103 tuberculosis patients of ADIH group were 17.5%, 26.2%and 27.2 % respectively, while those in 236 tuberculosis patients of control group were 29.7 % ,39.4 %and 40.7%, respectively (x2 =5.539, P＜0.05; x2 =5.458, P＜0.05; x2 =5.628, P＜0.05). The results of univariate analysis demonstrated that the risk of concurrent ADIH was significantly higher in patients with wild genotypes of CYP 2E1-7632T/A, CYP 2E1-1259G/C, CYP 2E1-1019C/T than in patients with other genotypes. After adjusted for sex, occupation and alcohol consumption status, the results of multivariate Logistic regression analysis also showed that wild genotypes of CYP 2E1-7632T/A, CYP 2El-1259G/C, CYP 2E1-1019C/T were significantly associated with higher risk of ADIH. The results of interaction analysis indicated that the wild genotypes of CYP 2E1-7632T/A and CYP 2E1-1259G/C or CYP 2E1-1019C/T had synergetic effects on the development of ADIH.Conclusions The risk of concurrent ADIH is significantly higher in patients with wild genotypes of CYP 2E1-7632T/A, CYP 2E1-1259G/C, CYP 2E1-1019C/T compared to patients with othergenotypes. Wild genotypes of CYP 2E1-7632T/A and CYP 2E1-1259G/C or CYP 2El-1019C/T have synergetic effects on the development of ADIH.

Objective To evaluate the relationship between serum level of hepatitis C virus (HCV) core protein and antiviral therapy response in patients with hepatitis C. MethodsOne hundred and sixty-two consecutive patients with chronic hepatitis C (CHC) were retrospectively analyzed. Serum level of HCV core protein and HCV RNA were measured. Among them, 35 CHC patients treated with pegylated interferon (PEG-IFN)+ ribavirin (RBV) were followed up before and at week 4, 24, 48 of treatment. The correlations between serum HCV core protein level, HCV RNA and antiviral therapy were evaluated. Comparison between groups was done by t test and comparison of paired data among groups was done by analysis of variance. Results Total 162 patients were divided into six groups according to the HCV virus load: 56 with HCV virus load≤1×103 IU/mL and HCV core protein absorbance (A)=0. 100±0. 029; 9 with 1×103 IU/mL＜ HCV virus load≤1 × 104 IU/mLand A=0. 246±0. 213; 11 with 1 × 104 IU/mL＜ HCV virus load≤1 × 105 IU/mL and A=0. 235±0.179; 28 with 1×105 IU/mL＜ HCVvirusload≤1×106 IU/mL and A=0. 422±0. 319; 51 with 1 × 106 IU/mL＜ HCV virus load≤ 1 × 107 IU/mL and A = 0. 603 ± 0. 330 ; 7 with 1 × 107 IU/mL＜HCV virus load≤ 1 × 108 IU/mL and A = 0. 900± 0. 379. The HCV core protein absorbance was positively correlated with HCV RNA level (r= 0.36, P＜0.05). The HCV core protein absorbance values of 35 CHC patients before therapy, at week 4, 24, and 48 of therapy were 0. 564 ±0. 296,0. 144±0. 062, 0. 091 ±0. 035 and 0. 112±0. 103, respectively. The HCV core protein absorbance values at week 4, 24, 48 all decreased significantly compared with that before therapy (t= 8. 563,9. 195, 9. 250; all P＜0.05), and there was significant difference between those at week 4 and week 24 (t=4. 301, P＜0.05). Conclusion Serum HCV core protein level is closely correlated with HCV viral load during HCV infection process and antiviral treatment.

Objective To analyze the genetic characterization(evolution, antigenicity, enzyme activity sites and glycosylation sites)of the neuraminidase(NA)gene of the novel influenza virus A/H1N1 in 2009 pandemic in Guangdong Province. Methods The viral RNA was extracted from 69 isolates of influenza virus A/H1N1 from patients in 2009 pandemic in Guangdong Province. NA gene fragments were amplified by reverse transcription polymerase chain reaction (RT-PCR) and sequenced. The other 52 NA gene sequences of influenza virus A in different years and different regions were retrieved from GenBank. The analysis of evolution and amino acid sequences were analyzed by MEGA 4.0 software. Results The homology of 2009 novel H1N1 influenza viruses in Guangdong and avian H5N1 influenza virus strains was high(＞85 % ). The amino acid distributions of potential antigenic sites were identical. The enzyme activity sites of NA genes of all virus strains were strictly conserved, which had eight glycosylation sites. But there were amino acid substitutions in 5 glycosylation sites, while it was identical with the 2001 avian H5N1 influenza virus. Conclusion The NA genes of 2009 novel H1N1 influenza viruses in Guangdong are high homologous with avian H5N1 influenza virus and the viral specific binding sites of neuraminidase inhibitor are not changed.

Objective To analyze clinical and laboratory features, viral load and viral shedding period of patients with mild or severe H1N1 influenza A infection. Methods Seventy mild cases and 16 severe cases with concurrent pneumonia were included from Shcnzhen area for analysis.Nasopharyngeal-swab specimens of patients were collected and viral load was detected by real-time quantitative polymerase chain reaction (PCR) assay during their hospitalization. The viral load and viral shedding period were compared between patients over 14 years old and less than 14 years old, and between 70 mild cases without pneumonia and 16 severe cases with pneumonia. The statistic analysis was performed using t test and chi square test. Results The most common symptoms and signs of the patients were fever, cough and enlargement of tonsils. However, the severe cases suffered more frequently from cough, dyspnea and high fever compared with the mild cases (x2 = 10. 9 and 14.3, respectively, t=3.65; both P＜0.01 ). The levels of white blood cell (WBC) count and alanine arninotransferase (ALT) of severe patients were both significantly higher than those of mild patients(t= 3.2, 2.4,respectively; both P＜0.05). The chest radiology of the severe cases showed interstitial pneumonia,mostly with ground glass image. The viral load of patients under 14 years was significantly higher than those over 14 years [(4.86± 1.23) lg vs (4. 17±0.89) lg; t=2.3, P＜0.05], and the viral shedding period of patients under 14 years was significantly longer than those over 14 years [(5.33±0. 49) d vs(3. 63±0.28) d; t=3.4, P＜0.01]. The severe patients also displayed significantly higher viral load and prolonged viral shedding period than the mild patients [(6. 36±1. 44) lg vs (4. 35±0.99) lg, t=6.1,P＜0.01; (5.75±1.77) d vs (4. 24±1. 96) d, t=3.2, P＜0.01]. Conclusion Age anddisease severity of patients with H1N1 influenza A infection are significantly associated with viral load and viral shedding period.

Objective To study the expression profile of microRNA (miRNA) and the roles in pathogenesis of acute liver failure in mouse model. Methods Eighty-five BALB/c mice were divided into four groups: 40 in model group of acute liver failure were intraperitoneally injected with Dgalactosamine (D-GalN) and lipopolysaccharides (LPS); 20 in D-GalN group were injected with DGalN only; 20 in LPS group were injected with LPS only; 5 in control group were injected with saline.Liver histology of mouse was observed at hour 0, 5, 7 of injection, and sera and liver tissues were collected at hour 0, 1, 3, 5, 7, 9 of injection. Meanwhile, levels of inflammatory factors [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] in serum and liver tissue were detected by realtime polymerase chain reaction (PCR). Lock nucleic acid (LNA)-based miRNA microarray technology was used to detect the expression profile of hepatic miRNA, and the expression of miRNA was verified by real time quantification-polymerase chain reaction (RT-PCR). Mouse macrophage Raw264.7 cells were induced by LPS in vitro and the expressions of miRNA at different time points were detected.The comparison of means among groups was analyzed using one way ANOVA and the correlation were analyzed by Pearson and Spearman correlation. Results Microarray analysis found that the expression profile of miRNA during the acute liver failure changed dramatically. There were 97 miRNA in model group changed significantly compared with control group (P＜0.01), including 21 up-regulated and 27down-regulated at hour 5 and 7 of injection. Furthermore, the expressions of miR 146a and miR-155were verified by RT-PCR and found they both increased progressively over time after injection.Correlation analysis showed that miR-155 was well correlated with both TNF-α and IL-6 expressions.It was further found that miR-146a and miR-155 were both up-regulated in activated Raw264.7 cells in vitro. Conclusions The expression profile of miRNA changes during acute liver failure in mouse model. Inflammation associated-miR-146a and miR-155 are both up-regulated significantly, which indicatcs that they may play an important regulatory role in pathogenesis of acute liver failurc.

Objective To understand the clinical characteristics of pediatric patients who developed H1N1 influenza A virus-associated pneumonia during the outbreak of H1N1 influenza A in Shanghai. MethodsA dcscriptivc study was done to analyze the clinical and epidemiologic characteristics of 30 hospitalized children who developed complicated pneumonia caused by H1N1 influenza A virus infection in 2009 in Shanghai. The comparison of medians was done using rank sum test and comparison of rates was done using exact chi-square test. Results Among thirty pediatric patients with H1N1 influenza A virus-associated pneumonia, the median age was 5.9 years old, five cases (16.7 %) had pre-existing medical conditions. Twenty cases (66.7 % ) had been exposed to the classmates or family membcrs with fever. All cases had fever and cough. Eleven cases (36.7 %00 ) had tachypnca and ten (33.3%) had wheeze. Eleven cases (36.7%) showed white blood cell (WBC)＜4.0 × 109/L and 2 (6. 7%) had thrombocytopenia. All patients had bilateral or unilatcral patchy infiltrates in the lung indicated by chest X-ray and four (13. 3%) had extensive infiltrates with the evidence of pulmonary edema. One (3. 3%) critically ill child with pneumonia, chest computed tomography scan revealed lung fibrosis 3 months and 9 months after illness onset. Three(10. 0%) cases had pneumomediastinum and subcutaneous emphysema. Six cases (20. 0%) were complicated with acute respiratory failure, three (10. 0%) with acute asthmatic attack and one (3. 3%) with encephalitis. All patients were treated with oseltamivir plus antibiotics and four required mechanical ventilation. All patients survived. The median duration of fever in group with oseltamivir given within 2 days of fever onset was statistically shorter than that in group with oseltamivir given 2 days after fever onset (2 days vs 5 days, Z= -8. 015, P<0. 01). Conclusions Both pre-school age and schoolage children may develop complicated severe respiratory diseases after H1N1 influenza A virus infection. Early initiation of oseltamivir may shorten the duration of fever and reduce the occurrence of severe complications.

Objective To analyze the clinical features of the hospitalized children with laboratory-confirmed influenza in Suzhou. Methods The demographic information, laboratory test results, clinical features, treatments and outcomes of the hospitalized children with laboratoryconfirmed influenza were collected retrospectively. The data were analyzed using chi square test,Cochran-Armitage trend test. Results Four hundred and eighty hospitalized children were diagnosed with laboratory-confirmed influenza during the period of 2005 to 2009. Among these cases, 414 were subtype A and 66 were subtype B. The positive rate was 2.66%. Four hundred and nineteen cases (87.29 %) were ≤ 5 years old. Most of the cases developed during winter (from December to April the next year) and summer (from July to August). The age and sex distribution didn't vary from year to year (x2=9. 7768,x2 = 8. 7573; both P＞0.05). The mean disease course was (16.22± 9.41)days, and the mean hospitalization duration was (7.89 ±2.97) days. The percentages of patients with symptoms of fever, dyspnea and diarrhea or requiring oxygen treatment decreased with age (Z =4. 9430, Z=2. 1021, Z=3. 2073 and Z=2. 3277, respectively; all P＜0.05). The percentages of cases with concomitant pneumonia and upper respiratory infection also decreased with age (Z =-3. 8762 and Z=-3. 5095, respectively; both P＜0.01). Fifteen point five percent (60/387 cases)of the cases were co-infected with pneumococcus and 15. 0% (72/480 cases) were co-infected with respiratory syncytial virus (RSV). The level of C-reactive protein was significantly higher in cases with bacterial co-infection than those with viral co-infection (Z= -3.1290, P ＜ 0. 01 ).Conclusions Hospitalized children with influenza are more common in winter and summer in Shuzhou.Many patients are co-infected with pneumococcus or RSV. The symptoms are more severe in younger children.