Changing trends in etiologies of hospitalized patients with liver disease during 2006 -2014 / 中华肝脏病杂志

Objective@#To analyze the changing trends in etiologies of hospitalized patients with liver disease and provide clinical basis for the formulation of medical policy.@*Methods@#Patients who were hospitalized in the Department of Infectious Diseases from 2006 to 2014 were selected as the research subjects. Data of patients with liver diseases were retrospectively analyzed to determine the proportion of main causes of infection, the proportion of different viral infections in viral hepatitis, and the changing trends in proportion of hepatitis B in different age groups. Kruskal-Wallis test was used for statistical analysis.@*Results@#During 9 consecutive years, the overall proportion of inpatients with liver disease decreased continuously, but the number of patients increased. The top five etiologies of liver diseases were viral hepatitis, drug-induced liver disease, autoimmune liver disease, alcoholic and non-alcoholic fatty liver disease. The proportion of viral hepatitis decreased gradually, and the proportion of drug-induced liver disease and autoimmune liver disease increased markedly. Among viral hepatitis patients, hepatitis B, hepatitis C and hepatitis E were in the top three, with hepatitis B stabilized at around 70%, and the proportion of hepatitis C showed an upward trend. The hospitalization time of hepatitis B patients was gradually shortened, the difference was statistically significant (χ 2 = 205.31, P < 0.001), and the hepatitis B patients were mainly distributed in age groups 31-40, 41-50, and 51-60, the total proportion was above 60%. The difference between the different years of the same age group was not evident, but the proportion of hepatitis B patients decreased gradually in the 14-23 –year- old age group, the difference was statistically significant (χ 2 = 19.51, P = 0.01).@*Conclusion@#Liver disease still holds a principal position in the distribution of infectious diseases, and especially the cause of non-infectious liver disease require sufficient attention and concern. The use of hepatitis B vaccine has effectively diminished the infection rate, but the prevention and control of chronic hepatitis B infection is still facing challenges.

Clinical analysis of twenty patients with hepatic echinococcosis in Nanjing City / 中华地方病学杂志

Objective To investigate the epidemiological and clinical features of patients with hepatic echinococcosis in Nanjing City. Methods Epidemiological data, clinical manifestations, laboratory and image examination results, diagnosis and treatment of twenty patients with hepatic echinococcosis from January 2009 to December 2016 at the First Affiliated Hospital of Nanjing Medical University were retrospectively analyzed. Results A total of 20 hospitalized cases were investigated, of which 7 cases were males and 13 cases were females, the median age was 48 years (17 - 76 years). One case came from Tibet which was the epidemic area of hepatic echinococcosis;the other 19 cases were local hepatic echinococcosis patients including 3 cases that had raised dogs for many years and 2 cases engaged in processing and selling livestock products. Among the 20 cases, 14 cases had no remarkable symptoms and not sought medical advice until imaging examination showed focal liver lesions during a routine check-up, 2 cases saw a doctor due to hepatic cyst, 1 case due to progressive enlarged hepatic hemangioma, the remaining 3 cases due to abdominal discomfort or pain. In addition, 7 cases were diagnosed definitely in 7 to 14 days, 4 cases in 15 to 30 days, 5 cases in > 30 to 60 days, 1 case in 3 months, 1 case in 7 years and 1 case in 9 years. Thirteen patients had partial hepatectomy; the other 7 patients received endocystectomy. All patients had a good prognosis during the follow-up period. Conclusions Most of the local hepatic echinococcosis cases have no definite epidemiological data and no significant manifestations, leading to misdiagnosis and missed diagnosis. Clinical medical staff should strengthen their understanding of imaging characteristics especially the sonographic features of hepatic echinococcosis.

Association between nuclear factor kappa-light-chain-enhancer of activated B cells genetic polymorphisms and HCV susceptibility among the Chinese population under high-risk / 中华流行病学杂志

Objective To explore the association between nuclear factor kappa-light-chainenhancer of activated genetic polymorphisms in B cells (NF-κB) and the HCV susceptibility,among the Chinese population.Methods A total of 1 679 participants were enrolled;including 503 drug users and 1 176 other participants at risk under the exposure for blood.By using the logistic regression analysis,related risk factors for HCV infection among subjects were analyzed.Two NF-κB pathway variants,NF-κB1 rs72696119 and REL rs13031237 were then genotyped by TaqMan assay method.Logistic regression analysis was performed to analyze the association between gene polymorphisms and the susceptibility on HCV.Results Among the drug users,women (OR=0.408,95％CI:0.308-0.767) appeared to be associated with the decreased risk for HCV infection,while factors as drug injection (OR=8.817,95％CI:5.577-13.937) and the duration of drug-intake ＞5.5 years (OR=2.891,95％CI:1.824-4.583) were associated with the increased risk for HCV infection.Among the participants who had been exposed to blood,women (OR=3.431,95％ CI:2.360-4.988) were associated with the increased risk for HCV infection,while the levels of education beyond elementary school (OR =0.613,95％ CI:0.429-0.876) were associated with the decreased risk for HCV infection.Compared to the reference NF-κB1 rs72696119 CC genotype,the carriage of GG genotype was associated with an increased risk of susceptibility on HCV (OR=1.412,95％ CI:1.035-1.927) among the total study population.Results from the interaction analysis showed that there was no interactive effects appeared between rs72696119 and route of infection,or between rs72696119 and gender among the total population under study (all P＞0.05).Conclusion NF-κB1 polymorphism rs72696119 and related factors seemed associated with the susceptibility to HCV infection among high-risk Chinese populations.

Association between nuclear factor kappa-light-chain-enhancer of activated B cells genetic polymorphisms and HCV susceptibility among the Chinese population under high-risk / 中华流行病学杂志

Objective To explore the association between nuclear factor kappa-light-chainenhancer of activated genetic polymorphisms in B cells (NF-κB) and the HCV susceptibility,among the Chinese population.Methods A total of 1 679 participants were enrolled;including 503 drug users and 1 176 other participants at risk under the exposure for blood.By using the logistic regression analysis,related risk factors for HCV infection among subjects were analyzed.Two NF-κB pathway variants,NF-κB1 rs72696119 and REL rs13031237 were then genotyped by TaqMan assay method.Logistic regression analysis was performed to analyze the association between gene polymorphisms and the susceptibility on HCV.Results Among the drug users,women (OR=0.408,95％CI:0.308-0.767) appeared to be associated with the decreased risk for HCV infection,while factors as drug injection (OR=8.817,95％CI:5.577-13.937) and the duration of drug-intake ＞5.5 years (OR=2.891,95％CI:1.824-4.583) were associated with the increased risk for HCV infection.Among the participants who had been exposed to blood,women (OR=3.431,95％ CI:2.360-4.988) were associated with the increased risk for HCV infection,while the levels of education beyond elementary school (OR =0.613,95％ CI:0.429-0.876) were associated with the decreased risk for HCV infection.Compared to the reference NF-κB1 rs72696119 CC genotype,the carriage of GG genotype was associated with an increased risk of susceptibility on HCV (OR=1.412,95％ CI:1.035-1.927) among the total study population.Results from the interaction analysis showed that there was no interactive effects appeared between rs72696119 and route of infection,or between rs72696119 and gender among the total population under study (all P＞0.05).Conclusion NF-κB1 polymorphism rs72696119 and related factors seemed associated with the susceptibility to HCV infection among high-risk Chinese populations.

Construction of the recombinant plasmids encoding nonstructural proteins of SFTS virus and immunogenicity study / 中华实验和临床病毒学杂志

Objective@#To study the construction and humoral immunogenicity of the recombinant plasmids encoding nonstructural proteins(NSs) of severe fever with thrombocytopenia syndrome(SFTS) virus(SFTSV).@*Methods@#Recombinant plasmids encoding NSs gene and optimized NSs gene of SFTSV were constructed by polymerase chain reaction(PCR) and cloned into eukaryotic expression vector pJW4303, which were named pJW4303-NSs and pJW4303-NSs-opt, respectively. Then, the plasmid pJW4303-NSs was tagged with Flag, named pJW4303-NSs-Flag. Meanwhile, Nhe I restrict site and tissue plasminogen activator(tPA) signal sequence were inserted to construct bi-optimized recombinant plasmid named pJW4303-tPA-NSs-opt. All plasmids were identified by sequencing. The transient expression of NSs was confirmed by Western blotting in human embryonic kidney 293T cells. The NSs-specific IgG antibodies in BALB/c mice which were immunized by intramuscular injection with electroporation were examined by enzyme-linked immunosorbent assay(ELISA).@*Results@#The recombinant plasmids pJW4303-NSs, pJW4303-NSs-opt, pJW4303-tPA-NSs-opt and pJW4303-NSs-Flag were successfully constructed. The expression of NSs was confirmed in lysates and supernatants of 293T cells. The NSs-specific IgG responses of all three recombinant plasmids were detected by ELISA in BALB/c mice. It was found that optimized recombinant plasmid pJW4303-NSs-opt elicited higher levels of the NSs-specific IgG than that of pJW4303-NSs in week 6, 8 which induced stronger immune response.@*Conclusions@#The recombinant plasmids encoding SFTSV NSs possess the satisfied immunogenicity. In addition, the plasmid pJW4303-NSs-opt could induce the strongest humoral immune response.

A retrospective study of clinical and immunologic features of 67 cases of patients with scrub typhus / 中华地方病学杂志

Objective To study the clinical and immunologic features of 67 cases of patients with scrub typhus.Methods Epidemiological data,clinical manifestations,laboratory and image examination results and treatment of 67 cases of patients with scrub typhus hospitalized from January 2010 to December 2013 at the First Affiliated Hospital of Nanjing Medical University were retrospectively analyzed.Results Autumn-winter type scrub typhus was predominant in the 67 patents.The main clinical manifestations included hyperpyrexia (100％,67/ 67),eschar or ulcer (91.0％,61/67),rash (52.2％,35/67),lymphadenopathy (46.3％,31/67) and other non-specific symptoms.Laboratory test results:①80.6％ (54/67) of the patients had normal or decreased white blood cell count,62.7％ (42/67) of them with elevated lymphocytes (＞ 40％),thrombocytopenia was detected in 34.3％ (23/67) of the patients,the lowest platelets count was 14 × 109/L.②14 of 17 cases (82.4％) had normal percentage of CD3+ T lymphocytes,CD8+ T lymphocytes increased in 16 of 17 cases (94.1％),while the percentage of CD4+ T lymphocytes (14 of 17,82.4％) and B lymphocytes (15 of 17,88.2％) decreased in most of the patients.③C reactive protein (CRP),ferritin and erythrocyte sedimentation rate (ESR) were elevated in 95.2％ (59/62),93.8％ (45/48) and 89.1％ (49/55) of the patients,respectively.④Elevated alanine aminotransferase (77.6％,52/67) and aspartate aminotransferase (80.6％,54/67) were frequent findings.⑤94.6％ (35/37) of the patients had increased adenosine deaminase.⑥ Antinuclear antibody was positive in 31.0％ (13/42) patients.⑦Epstein-barr virus DNA was detected in 15.6％ (7/ 45) cases,77.6％ (38/49) was accompanied with chest radiographic abnormalities.Two cases had tuberculosis.All the patients recovered after therapy with doxycycline,levofloxacin or moxifloxacin.Conclusion Autumn-winter scrub typhus has typical clinical manifestations in the 67 patients,and it can cause patients' immune disorders.

Viral load and cytokines in the pathogenesis of severe fever with thrombocytopenia syndrome / 中华传染病杂志

Objective To observe the relationship of viral load,serum cytokines and tissue damage after severe fever with thrombocytopenia syndrome virus (SFTSV)infection,and to explore the impact of SFTSV levels on tissue injury and prognosis.Methods Twenty-four ambulatory and hospitalized patients who were infected with SFTSV were enrolled between May 2011 and July 2012 at Department of Infectious Disease, First Affiliated Hospital with Nanjiang Medical University. According to their prognosis,they were divided into cure and death group,while 32 healthy blood donators were also enrolled from center blood station in Nanjing as control.The serum SFTSV load was detected using fluorescence quantitative polymerase chain reaction (PCR).The serum T helper (Th)1/Th2/Th17 cytokines in patients with severe fever with thrombocytopenia syndrome (SFTS)were determined dynamically and quantitatively by flow cytometry.The relationships between viral load,cytokines and serum enzymes, white blood cell (WBC),platelet (PLT)counts were analyzed.Comparisons among groups were achieved by rank sum test and correlation analysis among serum cytokines,blood cell counts and tissue damage was done by Spearman correlation test.Results All of the 24 patients showed a positive reaction to SFTSV RNA.The SFTSV loads of 21 cured cases,those of 2 were > 7.0 lg copy/mL,and those of 3 death patients were 6.7 lg copy/mL,8.8 lg copy/mL and 9.8 lg copy/mL,respectively.Serum level of interleukin (IL)-6 (21 .76 pg/mL in day 5 and 7.12 pg/mL in day 7)and IL-10 (14.33 pg/mL in day 5 , 14.13 pg/mL in day 7 and 3.01 pg/mL in day 9)of cured patients were significantly higher than those of healthy controls (IL-6:2.82 pg/mL and IL-10:1 .56 pg/mL)(P <0.05 ).At day 7 and day 9,serum levels of IL-6 of death cases were 137.61 pg/mL and 1 450.83 pg/mL,respectively and serum levels of IL-10 were 50.26 pg/mL and 49.43 pg/mL,respectively.Both of the indicators in the death group were significantly higher than those of cure group (P <0.05 ).However,serum levels of IL-2 and IL-4 were significantly lower than those in healthy control group (P <0.05 ).In the cure group,WBC and PLT counts were lowest during the early course of the disease,and serum alamine aminotransferase (ALT), aspartate aminotransferase (AST ), lactic dehydrogenase (LDH ) and creatine kinase (CK ) were significantly higher than their upper limits of normal.The correlation analysis showed that serum IL-6, IL-10 levels were negatively correlated with PLT count (r=-0.390 and -0.608,respectively;both P <0.01),and positively correlated with SFTSV load (r=0.560 and 0.758,respectively),ALT (r=0.412 and 0.390,respectively),AST (r = 0.686 and 0.764,respectively),LDH (r = 0.633 and 0.677, respectively)and CK (r =0.527 and 0.636,respectively)(all P <0.01 ).Conclusions SFTSV load, IL-6,IL-10 and serum enzyme levels are closely related to the severity of the disease.The inflammatory and anti-inflammatory cytokine storm after SFTSV infection may be involved in the immune pathological injury in patients with SFTS.

The investigation of adefovir dipivoxil antiviral clinical efficacy on chronic hepatitis B and its effect on peripheral blood T lymphocyte subsets / 中国生化药物杂志

Objective To investigate the clinical efficacy of adefovir dipivoxil in antiviral treatment of chronic hepatitis B and its effect on peripheral blood T lymphocyte subsets.Methods 40 cases of patients with chronic hepatitis B were selected from The Fourth People’s Hospital of Qinghai in October 2009 to June 2012,the patients were given the antiviral treatment of adefovir dipivoxil.Before treatment,3 months,6 months and 12 months after treatment,the changes of liver function indicators of alanine transaminase(ALT),aspartate transaminase(AST)and total bilirubin(TBiL),and hepatitis B virus-DNA(HBV-DNA)were recorded;the total effective rate was observed;the peripheral blood T lymphocyte subsets changes was tested and analyzed by flow cytometry.Results The ALT,AST and TBiL of the patients all improved good,and the indicators of ALT,AST and TBiL after 12 months decreased significantly than before treatment,the HBV-DNA testing of the patients recovered negative,the difference was statistically significant (P<0.05 );the total effective rate of hepatitis B patients is 92.5%;the peripheral blood T lymphocyte subsets has no significant improvement in patients after antiviral treatment for 3 months.After the treatment for 6 months and 12 months,the CD4 +T and CD4 +/CD8 +T were significantly higher than those before treatment,there was no significant changes in CD3 +T and CD8 +T before and after treatment.Conclusion Adefovir dipivoxil has a good clinical efficacy in antiviral treatment of chronic hepatitis B,and can significantly improve the patients’liver function,improve peripheral blood T lymphocyte subsets imbalance.

Dynamic changes of peripheral blood lymphocyte subsets in fever with thrombocytopenia syndrome patients / 中华检验医学杂志

Objective The aim of this study is to dynamically investigate peripheral blood lymphocyte subsets in fever with thrombocytopenia syndrome (SFTS) patients at different stages,to evaluate the influence of these changes in the infection process.Methods Case-control study was used in the research.Twelveconfirmedthrombocytopeniasyndromevirus ( SFTSV ) infectedpatientswere enrolled.According to SFTS prevention guide issued by Chinese Ministry of Health,these patients were divided into two groups,recovery group and death group.For each group,dynamic profiles of the CD3 + T cells,CD4 + helper T cells,CD8 + cytotoxic T cell and CD3 - CD16 + CD56 + natural killer cells were tested by flow cytometry.Meanwhile, the relationshipsbetween these dynamicchanges and liver function,leukocytes,and platelets were analyzed respectively.Two independent-samples t test was used to compare the difference of the peripheral blood lymphocyte subsets count between the SFTS patients and healthy control.Small sample was analyzed by Mann-Whitney U test.Results In the early stage of infection,Th cells in peripheral blood of recovery group were significantly reduced and Th/Tc ratio was reversed.On day 5,7,9 of post infection,Th cell counts in peripheral blood were (740.9 ± 6.4),(836.2 ± 272.3 ) and ( 1083.6 ± 319.7 ) cells/μl respectively,which were significantly lower than health control ( 1351.4 ± 295.1 ) cells/μl ( t value was -2.883,-4.235,-2.145 respectively,all P ＜0.05).Tc cell counts were significantly more than healthy controls (690.1 ± 194.8) cells/μl through the course,which were ( 1006.3 ±356.5),(1166.4±242.4),(1102.4±245.9),(991.3±205.1) and (886.5±154.5) cells/μl on day 7,9,11,13,15 of the course (t value was 3.312,5.661,4.574,3.874,2.382,all P＜0.05).NK cells were decreased significantly from the ninth day of the course.Associated with abnormal changes of cell subsets,WBC and PLT decreased significantly,and serum ALT,AST,LDH and CK etc.were higher than normal level.With the disease recovery,the abnormality above was gradually improved.In contrast,death cases showed significant decrease in T and Th cells compared with health control (P ＜ 0.05).On day 7,8,9 of the course,the counts of total T cell were (735.9 ± 359.9),(724.9 ± 125.9),(845.3 ± 389.3) cells/μl and the counts of Th cell were ( 533.2 ± 246.9 ),( 532.1 ± 105.7 ),( 551.7 ± 86.9 ) cells/μl,significantly lower than healthy control ( 1727.9 ± 230.2 ) cells/μl and ( 1351.4 ± 295.1 ) cells/μl,with statistically differences (z value was - 2.828, - 2.342,- 2.342 and - 2.828, - 2.342, - 2.342,all P ＜ 0.05 ).On day 7,8,9 of the course,the numbers of NK cell in death group were ( 1141.8 ± 415.5),( 1047.2 ±68.4),( 1276.3 ±545.3) cells/μl,which were significantly more than health group (470.7 ± 242.2) cells/μl,with statistically differences (z value was - 2.180,- 2.335,- 2.258,all P ＜0.05).Conclusions SFTSV infection can induce cell immunity damage.The changes of lymphocyte subsets are associated with clinical classification and prognosis.Significant reduction of T cell and CD4 +cell in peripheral blood are accompanied with significant increase of NK cell,which may be a pivotal indicator of poor prognosis and play an important role in making appropriate strategy in clinical treatment.( Chin J Lab Med,2012,35:826-831 )

The mechanism of quinolone resistance in Pseudomonas aeruginosa / 中华临床感染病杂志

Objective To investigate the mechanism of quinolone resistance in Psendomonas aeruginosa.Methods The minimum inhibitory concentrations (MICs)of ciprofloxacin and levofloxacin with and without carbonylcyainde-m-chlorophenylhydrazone(CCCP)were determined by agar dilution method.Polymerase chain reaction(PCR)and DNA sequencing were used to study the mutations in quinolone resistance-determining region of gyrA and parC genes.The strains were genotyped by enterbacterial repetitive intergenie consensus-PCR(ERIC-PCR).Results Sixteen quinolones-resistant Pseudomonas aeruginosa strains were obtained.The MICs of ciprofloxacin and levofloxacin were not reduced significantly by adding CCCP.Thr-83→Ile of gyrA and Ser-87→Leu of parC were found simultaneously in 16 strains of resistant Pseudomonas aeruginosa.Analysis of ERIC-PCR products indicated that 16 quinolone-resistant strains had an identical band pattern which was different from that seen in the sensitive strains.Conclusion Mutations in gyrA and parC may be the main mechanism of quinolone resistance in clinical isolates of Pseudomonas aeruginosa.

The association between antigen-specific cytotoxic lymphocytes response and different clinical status in patients with hepatitis B / 中华传染病杂志

Objective To analyze human leucocyte antigen (HLA)-A0201 restricted antigen-specific cytotoxic lymphocytes (CTL), and to investigate the difference of T cell response to specific antigen epitopes between patients with acute phase of acute hepatitis B and active phase of chronic hepatitis B. Methods Peripheral blood mononuclear cells (PBMC) from 5 patients with acute phase of acute hepatitis B and 6 patients with active phase of chronic hepatitis B were isolated. The numbers and functions of CD8+ T-lymphocyte epitope peptide specific CTL were detected using enzyme-linked immunosorbent spot (ELISPOT) assay, and the 3 peptides were from HBV polymerase region (Pol575-583), envelope region (Env348-357) and core region (Core18-27), respectively. The data were analyzed using t test. Results The spot formation cell counts (SFC) of Pol575-583, Env348-357 and Core18-27 stimulations in patients with acute phase of acute hepatitis B were 110±13, 165±17 and 185±20, respectively; and those in patients with active phase of chronic hepatitis B were 22±4, 23±5 and 30±5, respectively; the differences were all significant (t=10.9, 15.2 and 8.0, respectively, all P<0.05). The CTL responses to the three peptides in patients with acute phase of acute hepatitis B were Pol575-583<Env348-357<Core18-27; and the difference between responses to Pol575-583 and Core18-27 was significant (t=4.0, P<0.05), while there was no statistical difference between CTL responses to Env348-357 and Core18-27 (P>0.05). The SFC were increased upon non-antigen specific HLA-A2404 restricted epitope (Core117-125), but the difference was not significant compared with negative control group (P>0.05). Conclusions Hepatitis B virus-specific CTL responses in patients with acute hepatitis B are significantly higher than those in patients with chronic hepatitis B. The number and function of polyclonal CTL are both impaired in patients with chronic hepatitis B.