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Objective@#To analyze the distribution of HCV genotype in eastern Zhejiang Province and its correlation with sex, age, viral load, antiviral effect and so on.@*Methods@#A total of 501 cases of HCV infection seen in Ningbo No. 2 hospital from January 2011 to April 2018 were included. The HCV genotypes and HCV RNA were detected by gene chip method and RT-PCR respectively. The liver function and blood routine tests were performed and the APRI index was calculated. The factors affecting the SVR were analyzed for the patients who were partially treated with pegylated interferon and ribavirin (PR).@*Results@#The HCV genotypes of 501 cases were 1b、6、2a、3a、3b、1a from the higher to lower ranks, and genotype 1b was more than 50%.The distribution of HCV genotypes in different age groups was significantly different (χ2=95.433, P<0.001). The age of patients with 1a, 1b and 2 A was significantly higher than that of 3a, 3b, and type 6 (F=11.879, P<0.001). There were significant differences in viral load, AST, PLT and APRI index among different genotypes (F=2.920, χ2=12.400, F=6.294, χ2=12.700, all P<0.013), but ALT had little difference (χ2=7.994, P>0.157); 179 patients with PR standard regimen were followed up. The result showed that there was no significant difference in the overall efficacy of different genotypes (χ2=6.598, P=0.252), but the rate of sustained virological response in type 3 A was significantly higher than that of the 1b type (χ2=4.193, P=0.041).@*Conclusions@#The HCV genotypes are mainly 1b and 6 in eastern Zhejiang Province. There were significant differences in age, viral load, AST, PLT and APRI indices among patients with different HCV genotypes, and the therapeutic effect of PR regimen was better for genotype 1b than that of 3a.
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Objective@#To explore the effect of hepatitis C virus (HCV) on the expression of serine protease inhibitor Kazal1 (SPINK1) and its clinical implication.@*Methods@#mRNA and protein expression of SPINK1 in Huh7.5.1 cells infected by HCV JFH-1 and the control cells were measured by RT-PCR and western blotting, SPINK1 levels in the cell supernatants and sera of HCV patients were measured by enzyme-linked immunosorbent assay (ELISA), the difference of SPINK1 levels between healthy controls and HCV patients was analyzed.@*Results@#Expression of SPINK1 mRNA and protein was higher in Huh7.5.1 cells infected by HCV JFH-1 than in the control cells, serum SPINK1 levels was much higher in HCV patients than in healthy controls (P=0.016).@*Conclusions@#HCV can upregulate the expression of SPINK1.
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Objective To investigate the clinical value of serum apolipoprotein A-1 (Apo A-1) detection in HBV-related liver cancer.Methods Totally 362 cases of patients with chronic HBV infection were enrolled from January 2010 to December 2014 in our hospital,including 88 cases of chronic hepatitis B,94 cases of HBV-related liver cirhosis,18 cases of HBV-related liver cancer (without cirrhosis) and 162 cases of liver cirrhosis merged cancer.At the same time,45 cases of healthy people were selected for normal control.The serum Apo A-1,AFP and other laboratory markers were detected,and the test results were statistically analyzed.Results The difference of Serum Apo A-1 and AFP levels in all groups was statistically significant (F =29.86,x2 =112.53,P =0.000).As the disease progressed,the serum levels of Apo A-1 gradually decreased (P ＜ 0.05).But the difference of Apo A-1 level between normal control and HBV-related liver cancer group (without cirrhosis),chronic hepatitis B and liver cirrhosis merged cancer group,liver cirrhosis and liver cirrhosis merged cancer group was not statistically significant (all P ＞ 0.05).The liver cancer patients with Child-Pugh score A,B,C had different serum Apo A-1 levels (all P ＜ 0.05);The serum Apo A-1 level of liver cancer patients with Child-Pugh score A was significantly higher than that of liver cirrhosis (t =-3.02,P =0.003),but the differences of serum Apo A-1 levels between liver cancer and liver cirrhosis patients with Child-Pugh score B and C were not statistically significant (t =0.52,1.19,P =0.610,0.240).The serum Apo A-1 levels of liver cancer patients with TNM stage Ⅰ and Ⅱ were significantly higher than those with TNM stage Ⅲ and Ⅳ (t =3.85,P ＜ 0.001).Conclusion The serum Apo A-1 levels of HBV-related liver cancer patients are related with cirrhosis,Child-Pugh score and TNM stage,and the liver reserve function,the body's stress response and many other factors may contribute to the expression of serum Apo A-1.
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Objective To identify antimicrobial susceptibilities of community-acquired Staphylococcus aureus infections and the risk factors of severe infections.Methods Clinical data of 184 cases of community-acquired Staphylococcus aureus infections collected from 4 hospitals in Ningbo during May 2008 and May 2013 were reviewed.Microbial sensitivity test and virulence genes ( pvl and tst) detection were performed in clinical isolates, and SCCmec genotyping was performed in methicillin-resistant Staphylococcus aureus ( MRSA) strains.Binary logistic regression analysis was used to identify the risk factors for severe infections.Results Among 184 cases of community-acquired Staphylococcus aureus infections, 39 ( 21.20%) were severe cases. Staphylococcus aureus strains were highly resistant to penicillin, erythromycin and clindamycin, but more than 75% strains were sensitive to oxacillin, aminoglycosides, quinolones, rifampicin and vancomycin.Logistic regression analysis showed that advanced age (OR=1.024, 95%CI:1.005-1.043, P<0.05), malignant tumor (OR=15.288, 95%CI:1.609-145.229, P<0.05) , autoimmune diseases or long-term hormone therapy ( OR=12.102, 95%CI:2.082-70.338, P <0.01 ) were risk factors for severe community-acquired Staphylococcus aureus infections. Conclusions Strains isolated from the patients with community-acquired Staphylococcus aureus infections in Ningbo are usually sensitive to oxacillin, aminoglycosides, quinolones, rifampicin and vancomycin, which may be recommended for clinical use.Elder patients and those with malignant tumor, autoimmune diseases or long-term hormone therapy are more likely to develop severe Staphylococcus aureus infections.
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Objective To investigate hepatitis B virus (HBV) gene mutations related to entecavir (ETV)-resistance in patients with chronic HBV infections.Methods Serum samples were collected from 44 patients with chronic HBV infections and resistant to ETV treatment who were admitted in Ningbo No.2 Hospital during February 2010 and May 2014.The HBV polymerase regions were amplified by real-time fluorescent quantitative polymerase chain reaction (PCR) method,and the PCR products were analyzed with direct sequencing.SPSS 16.0 was used to assess the frequency of HBV polymerase gene mutations,and its relation to the viral genotype and clinical features.Results The most common HBV polymerase gene mutation was rtS202G/I (52.28％,23/44),followed by rtT184A/G/I/S (36.36％,16/44) and rtM250V/L (11.36％,5/44).Nine mutation patterns were detected,in which rtL180 + rtM204V + rtS202G/I (38.64％,17/44) and rtL180 +rtM204V + rtT184A/G/I/S (27.27％,12/44) were the most frequent ones.The difference in gene mutations between genotype B and C was of statistical significance (x2=12.294,P ＜0.01).Patients carrying rtT184A/G/I/S mutations were associated with worse liver function (x2 =14.499,P ＜ 0.01),and those carrying rtM250V/L mutations were associated with lower HBeAg positive rate (x2 =10.057,P ＜ 0.01).Conclusions rtL180M + rtM204V + rtS202G/I is the most common HBV polymerase gene mutation related to ETV resistance in patients with chronic HBV infections.Different gene mutations may be associated with HBV genotypes,severity of liver damages,and HBeAg positive rate.
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Objective To explore the clinical efficacy and adverse reactions of transarterial chemoembolization(TACE)combined with sorafenib for the treatment of liver carcinoma.Methods Forty-eight cases of intermediate-advanced liver carcinoma patients were divided into TACE combined with sorafenib group(test group)and TACE only group(control group)according to the wishes of the patient,with 24 patients in each group.The median Survival Time(mOS),clinical efficacy,quality of life,liver function indexes and adverse reactions were compared in two groups.Results Until the deadline of follow-up time,the mOS in test group(15.9 months)was significantly higher than that in control group(9.3 months).The difference was statistically significant(P<0.05).The ORR,CBR in test group(58.3%,87.5%)were significantly higher than those in control group(33.3%, 54.2%).The difference was statistically significant(P<0.05).The quality of life improvement rate in test group(79.2%)was significantly higher than that in control group(37.5%).The differences were statistically significant (P<0.05 ).The ALB,TBIL increased significantly after treatment than before treatment in both groups.The differences were statistically significant(P<0.05 ).Conclusion TACE combined with sorafenib is more efficiency than TACE only treatment,and it could effectively extend the period of life to guarantee the quality of life.What's more,it has well tolerated adverse reactions,which is worthy of promoting.
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Objective To evaluate the clinical application of liver shear wave velocity (SWV) in comparison with common serum score systems [AST to platelet ratio index (APRI),Forns,S index and FIB-4] in assessment of liver fibrosis in patients with chronic hepatitis.Methods A total of 237 chronic hepatitis patients with liver fibrosis confirmed by liver biopsy,who were admitted in Ningbo Second Hospital during October 2010 and April 2013,were enrolled in the study.Liver shear wave velocity were measured by acoustic radiation force impulse (ARFI),and the score of APRI,Forns,S index and FIB-4 were calculated based on the measurement of serum markers.Liver fibrosis stages were classified as S0-S4 according to the Scheuer scoring system,and stages ≥ S2 were identified as significant liver fibrosis.The diagnosis value of SWV,4 common score systems and their combination for significant liver fibrosis was evaluated by receiver operating characteristic curve (ROC).Results A significant linear correlation was found between SWV and the stage of fibrosis (r =0.46,P ＜ 0.01).The areas under the ROCs of SWV and 4 common score systems (APRI,Forns,S index and FIB-4) for the diagnosis of significant liver fibrosis were 0.758 (0.696-0.821),0.727 (0.662-0.793),0.777 (0.717-0.836),0.747 (0.684-0.810) and 0.737 (0.673-0.802),respectively.The area under the ROC of the combined prediction nodel established with Logistic regression was 0.810.Conclusion Liver shear wave velocity measured by ARFI is of clinical value in noninvasive assessment of liver fibrosis,and the prediction accuracy can be improved when it is combined with other noninvasive indices.
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Objective To investigate the efficacy of polyethylene glycol (PEG)-interferon α (PEG-IFNα) in treating HBeAg-positive chronic hepatitis B (CHB) and explore the relationship between hepatitis B virus (HBV) genotypes and the effect of interferon α (IFNα) therapy.Methods A total of 199 CHB patients with known genotypes were given subcutaneous injection of PEG-IFNα-2a or PEG-IFNαt-2b once a week for 48 weeks,with another 24 weeks follow up.The seroconversion of HBeAg influenced by HBV genotypes were analyzed after discontinuation of treatment.Results In local area,genotype C was the major genotype[64.32％ (128/199)].Except serum ALT and AST level,the differences in gender,age,liver inflammation,degree of liver fibrosis,HBeAg level and HBV DNA level between genotype B and C were not statistically significant(all P ＞0.05).The seroconversion rate of HBeAg in patients with genotype B at early stage of therapy (3 months) was significantly higher than that of patients with genotype C [26.76％ (19/71) vs 10.16％ (13/128),x2 =9.330,P =0.002].While at the end of follow-up,seroconversion rate of HBeAg in patients with genotype B (followed up for 6 months) was higher than that of patients with genotype C [39.44％ (28/71) vs 30.47％ (39/128)],but the difference was not statistically significant(x2 =1.645,P =0.200).By univariate analysis based on log-rank test,the time of HBeAg seroconversion in patients with genotype B was much earlier than that of genotype C [(13.99 ± 0.67) months vs (15.47 ± 0.41)months],but the difference was not statistically significant (P =0.150).Conclusions The seroconversion rate of HBeAg in patients with genotype B treated with PEG-IFNα was significantly higher than that of genotype C in early stage of therapy (3 months),while similar at the end of therapy.
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Objective To evaluate HBeAg quantification in predicting the efficacy of pegylated interferon (PegIFN) α treatment for patients with HBeAg-positive chronic hepatitis B (CHB).Methods A total of 216 HBeAg-positive CHB patients admitted in Ningbo No.2 Hospital during March 2009 and December 2011 were enrolled in the study.All patients were given subcutaneous injection of PegIFNα-2a or PegIFNα-2b weekly for 48 weeks and followed up for 24 weeks after discontinuation.Patients were divided into HBeAg seroconversion group and non-seroconversion group at the end of the follow-up.Receiver operating characteristic (ROC) curves were used to evaluate HBeAg levels at baseline and 12,24 weeks of treatment in predicting HBeAg seroconversion.Results HBeAg seroconversion was observed in 31.48％ (68/216) patients at the end of follow-up,and there was no significant difference in seroconversion rate between patients treated with PegIFNα-2a and those with PegIFNα-2b (32.00％ vs.29.27％,P ＞ 0.05).There was significant difference in baseline HBeAg levels between patients with HBeAg seroconversion and those without HBeAg seroconversion (Z =-3.834,P ＜ 0.05).HBeAg seroconversion patients had a tendency of rapidly decreasing HBeAg level,but there was no significant difference in decreasing rate between seroconversion and non-seroconversion patients (F =3.321,P ＞ 0.05).ROC curves showed that HBeAg level at 24-week was the best indicator for predicting HBeAg seroconversion with area under curve of 0.861.Conclusion Serum HBeAg level at 24-week of treatment may be used to predict the HBeAg seroconversion in HBeAg-positive CHB patients treated with PegIFNα.
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Objective To comparatively analyze the immunological characteristics of patients with mild and severe influenza A (H1N1), and to provide the evidence for condition monitoring and treatment . Methods 52 cases with mild influenza A ( H1N1), 152 cases with severe influenza A ( H1N1) and 26 healthy subjects from July 1, 2009 to December 31, 2009 were enrolled in the study.Lymphocyte subsets in peripheral blood were analyzed by flow cytometry and the serum concentrations of interferon -γ( IFN-γ) and interleukin-4 (IL-4) were detected by enzyme-linked immune-sorbent assay (ELISA).Results The total lymphocyte counts were decreased obviously in patients with severe influenza A ( H1N1) than in mild pa-tients and in healthy subjects (P0.05).Con-clusion Immune dysfunction in patients with influenza A (H1N1) infection is associated with the severity of disease, especially cellular immunity .Therefore, monitoring of the immune system is valuable for the diag-nosis of influenza A(H1N1) infection.
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Objective To explore the effect of hepatitis B virus(HBV) on the expression of apolipoprotein A1 (ApoA1) and its regulatory mechanism.Methods RT-PCR and Western blot were used to measure the expression of ApoA1 in HepG2 and HepG2.2.15 cells,serum ApoA1 and high density lipoprotein cholesterol(HDL-C) levels in patients with HBV infection and in healthy individuals were measured by biochemical analyzer,statistical difference was analyzed by SPSS13.0,HepG2 cells was co-transfected with ApoA1 promoter containing the luciferase gene and HBV infectious clone pHBV1.3,luciferase activity was measured,expression of ApoA1 in HepG2 cells was measured by RT-PCR and Western blot after transfected with pHBV1.3.Results Expression of ApoA1 mRNA and protein was lower in HepG2.2.15 cells than in HepG2 cells,serum ApoA1 and HDL-C levels were much lower in HBV patients as compared to healthy individuals( P＜0.05 ),HBV represses ApoA1 gene promoter activity,ApoA1 mRNA and protein expression in HepG2 cells.Conclusion HBV can inhibit the expression of ApoA1 bothin vivo and in vitro.
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Objective To investigate the relationship between HBV polymerase gene mutations and HBV genotypes in chronic hepatitis B (CHB) patients resistant to adefovir dipivoxil (ADV).Methods Blood samples were collected from 114 ADV-resistant CHB patients during February 2010 and May 2012.The HBV polymerase regions from serum samples were amplified with real-time PCR,and the PCR products were sequenced.Normal distribution data were presented as x ± s,and non-normal distribution data were presented as M (P25-P75) ; for homogeneous data analysis of variance and LSD-t test were performed.Results There were 8 types of HBV polymerase gene mutations in 114 CHB patients; single point mutation was detected in 102 patients (89.47％) and double or triple points mutations were detected in 12 patients (10.53％).rtA181V/T/S (57.89％),rtN236T (14.91％) and rtA181V/T/S + N236T (9.65％) were the predominant mutations.For 21 patients (18.42％) with HBV genotype B,rtN236T mutation was prevalent (47.62％,10/21) ; while for those with HBV genotype C (93,81.58％),rtA181V/T/S mutation was the predominant (65.59％,61/93).The differences of rtA181V/T/S (x2 =12.269,P ＜0.01) and rtN236T (x2 =18.658,P ＜0.01) mutation rates between B and C genotypes were statistically significant.Conclusion rtA181V/T/S,rtN236T and rtA181V/T/S + rtN236T are the major HBV polymerase gene mutation types in ADV resistant CHB patients,and mutation types are related with HBV genotypes.
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Objective To explore the effect of hepatitis B virus(HBV) on the expression of high sensitive C-reaction protein(hs-CRP) and its clinical implication.Methods mRNA expression of hs-CRP in HepG2 and HepG2.2.15 cells was measured by RT-PCR,serum hs-CRP levels in patients with HBV infection and in healthy individuals were measured by biochemical analyzer Olympus5400,the expression of hs-CRP difference among patients with chronic hepatitis B,liver cirrhosis and hepatocellular carcinoma were analyzed.Results Expression of hs-CRP mRNA was higher in HepG2.2.15 cells than in HepG2 cells,serum hs-CRP levels was much higher in HBV patients as compared to healthy individuals ( P＜0.05 ),hs-CRP was detected at higher levels in patients with liver cirrhosis or hepatocellular carcinoma than those with chronic hepatitis B.Conclusion HBV can upregulated the expression of hs-CRP,which is associated with the disease progression.
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Objective To investigate the pathogens and their antibiotic resistance profile of hospital and community-acquired spontaneous bacterial peritonitis ( SBP) in patients with liver cirrhosis. Methods Eighty-four cirrhotic patients with SBP were recruited, in which 61 (72. 6% ) were diagnosed as communityacquired SBP and 23 (27.4% ) were diagnosed as hospital-acquired SBP. Bacterial identification and drug susceptibility tests were performed. SPSS 16. 0 was used for statistical analysis. Results There were 68 (81.0%) Gram-negative strains and 16 (19.0%) Gram-positive strains. Escherichia coli and Klebsiella pneumoniae were the top two prevalent strains. In community-acquired group, there were 21 (21/61, 34. 4% ) strains of Escherichia coli and 15 ( 15/61 , 24. 6% ) strains of Klebsiella pneumoniae; while in hospital-acquired group, it was 12 (12/23, 52.2%) strains and 6 (6/23, 26. 1%) strains, respectively. There were no statistical differences in the infection rates of Escherichia coli and Klebsiella pneumoniae between community and hospital-acquired SBP patients (x2 = 2. 21 and 0. 02, P > 0. 05). Drug susceptibility tests showed that Escherichia coli and Klebsiella pneumoniae were 100% sensitive to impenem, and the sensitivity to Piperacillin/Tazobactam was also high. But these strains were highly resistant to Ampicillin and Ampicillin/Sulbactam. All extended spectrum β-lactamases ( ESBLs) positive strains were resistant to cephalosporins, while ESBLs-negative strains were all sensitive to cephalosporins. Conclusions SBP in patients with liver cirrhosis are mainly caused by Gram-negative strains, especially Escherichia coli and Klebsiella pneumoniae. ESBLs-positive strains are highly resistant to cephalosporins, so proper use of right antibacterial agents is important.