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06 August 2012, Volume 27 Issue 08 Previous Issue    Next Issue

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Investigation into the correlations between T cell subsets and replication， liver function and pathology in children with hepatitis B virus.  LEI Xiao-yan*，SUN Yong-hong， YANG Ju-lan，YUAN Hong. *Department of Pediatrics， 2012, 27(08): 591-594.  Abstract ( )   Objective    To investigate  the correlations between T cell subsets and HBV  replication  levels， liver function and liver pathology in children with hepatitis B virus. Methods    Outpatient and inpatient treatment according to A total of 124 cases of HBV-DNA positive hepatitis B were included in the study. The Department of Pediatrics in The People’s Hospital of Gansu Province and Institute of Infectiou Diseases the First Hospital of Lanzhou University from April 2008 to April 2010 outpatient and hospitalized children with chronic hepatitis B， Among the 124 patients， 84 were male， 40 female；65 were HBV carriers， 59 chronic hepatitis（31 mild，18 moderate and 10 severe）；peripheral blood T cell subsets， cccDNA， DNA， hepatitis B virus markers and liver function were detected； histopathological  examinations were  performed  in  46 cases. Mean while 60 cases of HBsAg (-) were included as the normal control group. Results    In each group of children with hepatitis B，peripheral blood CD3+， CD4+and CD4+/CD8+ were significantly lower than the control group( F = 13.26、12.34、14.73，P＜ 0.01)， CD8+ higher than the control group（F = -11.87，P＜0.01）； in carriers group and mild group CD3+， CD4+and CD4+/CD8+ were significantly lower than the moderate group and severe group， differences being statistically significant. In the moderate and severe groups， HBV cccDNA positive rate was higher than that in the HBV carriers or the mild group（F = 25.429，P＜0.01）. HBVcccDNA positive group and HBeAg positive group were compared with HBVcccDNA negative group and HBeAg negative group， and CD3+， CD4+and CD4+/CD8+ were reduced，while CD8+ cells were increased， while was statistically significant . CD8+ cells increased with increasing viral load（F =-11.43  P＜0.01），while CD3+， CD4+and CD4+/CD8+ decreased with the increase（F＝12.42，13.87，12.75，P ＜0.01）. Abnormal transaminases group was compared with normal group， and CD3+， CD4+and CD4+/CD8+ were reduced (t＝15.28，14.63，27.74，P ＜0.01），while CD8+ cells were increased（t =-16.39，P ＜0.01）， which was statistically significant. CD3+， CD4+and CD4+/CD8+ increased with the degree of inflammation，and fibrosis tended to increase（t＝9.25，11.85，26.34，P＜0.01），and CD8+ increased with the degree of inflammation，and fibrosis showed a downward trend（t =-10.18，P ＜0.01）. Conclusion    T-cell immune dysfunction and the level of viral replication and liver function and liver pathology have a certain relationship. HBV active replication increases immune disorders and immune function recovery，which remoues virus and at the same time leads to pathological changes in liver tissue.

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Pathological observation of acute necrotizing encephalopathy associated with 2009 influenza A（H1N1） virus in chidren. CAO Juan， LI Cheng-rong， WEN Fei-qiu， DENG Ji-kui， WANG Guo-bing， ZHENG Yue-jie 2012, 27(08): 595-598.  Abstract ( )    Objective    To investigate possible immunopathogenesis of acute necrotizing encephalopathy （ANE） associated with 2009 influenza A （H1N1） virus through observing pathological changes in children brain tissues. Methods Three children died from ANE were included in this study. Hematoxylin—Eosin staining， myelin staining，microglia paraffin section staining and immunohistochemical EliVision method were performed， respectively. Results    Processus spinosus of microglia in brain tissues of ANE was observed to disappear and become round. Myelin staining confirmed that nerve fibers of central nervous system changed to demyelination. It was confirmed that microglia was activated in ANE by the Nao menke and Feigin staining method. Immunohistochemisty showed that activated microglia expressed TNF-α and IFN-α. No infiltration of lymphocytes and neutrophilic granulocyte were obseved in brain tissues of ANE chidren. Conclusions    The abnormal hyperplasia of microglia might be one of the pathogenesis involved in the ANE.

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Combied antifungal treatment for invasive fungal infection in children with hematological diseases.    2012, 27(08): 599-605.  Abstract ( )   Objctive    To evaluate the ef?cacies of treating invasive fungal infection （IFI）with double combinations in children with hematological diseases. Methods    A retrospective clinical study was made on curative effect based on 39 patients from January 2006 to April 2011.Results    A total of 48 IFIs were  treated with  combined antifungal therapy. Among them， 11 had possible diagnosis，32 probable diagnosis and 5 proven diagnosis. Eighteen IFIs received amphotericin B combined with caspofungin， 3 invalid； 13 received amphotericin B and itraconazole injection，4 oral  solution，all effective； 9 received amphotericin B and voriconazole， 1 invalid； 6 received caspofungin and itraconazole， 1 invalid； 5 received caspofungin and voriconazole，all effective. Totally 27 responded with complete remission，8 partial remission，1 abandoned，4 dead. There was no significant difference among triazole combined with amphotericin B or caspofungin and amphotericin B with caspofungin （P＞0.05）.Conclusion    Combined antifungal therapy shows impressive curing activity on patients with severe impaired immunological function，who receive strong chemotherapy，HSCT with past history of IFI，severe or breakthrough IFI，monotherapy failure or have intolerable side effects，and those having graft-versus- host disease after HSCT.  Triazole combined with amphotericin B or caspofungin and amphotericin B with caspofungin are all choice.

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CIinical features and antimicrobial resistance of invasive pneumococcal disease in 38 children.    WANG Man-li*，YAN Yong-dong，HUANG Li，ZHU Can-hong，WANG Yu-qing，CHEN Zheng-rong，Z 2012, 27(08): 604-607.  Abstract ( )   Objective    To investigate the clinical features of invasive pneumococcal disease（IPD）and its antimicrobial resistance in Suzhou，and to raise the level of diagnosis and treatment of this disease. Methods    The clinical data of 38 cases of IPD admitted to the Children's Hospital Affiliated to Soochow University from January 2004 to march 2011 and antimicrobial resistance were analyzed retrospectively. Results    （1）The 38 cases of IPD were identified，among whom 25 were male and 13 female，the ratio being 1.92:1. The ages ranged from 78 days to 12 years；18 cases（47.37％） were aged less than 2 years，and 12 cases （31.58％）were aged from 2 to 5 y，8 cases（21.05％） were aged older than 5 years．Among the 38 children，25 patients（65.79％） had sepsis， the most common clinical manifestation ，of whom 10 cases were complicated with pneumonia；12 patients （31.58％） had pneumococcal meningitis，of whom 4 cases were complicated with pneumonia；2 patients（5.26％） had necrotizing pneumonia；2 patients had sepsis ，meningitis and pneumonia.Six cases（15.79％）had an underlying disease；6 cases（15.79％）were with combined other pathogen infections. of the patients 34  were cured，2 died and 2 abandoned therapy．（2） We studied drug resistance of all the invasive pneumococcus：52.63％strain were not sensitive to penicilin，and the resistance to the other 9 commonly used antibiotics was erythromycin（94.74％），clindamycin（93.75％），tetracycline（82.25％），trimethoprim-sulfamethoxazole（61.54％），cefotaxime（23.53％），chloromycetin（15.79％），levofloxacin（0），vancocin（0），and rifampicin（0）．The multidrug resistance rate was 86.21％. Conclusion    Invasive pneumococcal disease has the highest incidence in children under 2 years old. Sepsis and purulent meningitis are the most common clinical manifestation. It is very important to monitor the resistance of invasive pneumococcus for clinical use of medicine．

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Detection of human bocavirus in children with acute wheezing and the relevant analysis about its viral load.  WANG Ting ， LI Chang-chong ，CHEN Xiao-fang， NIE Ying， CHONG Lei. Department of R 2012, 27(08): 608-611.  Abstract ( )   Objective  To investigate the epidemiologic features of human bocavirus （HBOV） in children with acute wheezing in Wenzhou area and analyze the correlation between disease severity and viral load. Method Between March and August of 2011，237 nasopharyngeal aspirates （NPA）were collected from children who had been hospitalized for acute expiratory wheezing in Wenzhou Yuying children's hospital. During the same period， 96 control specimens were obtained from children hospitalized without wheezing. HBOV was assessed in all samples by real-time PCR method. Results HBOV was detected in 62 （26.2%）NPA samples from 237 acute wheezing children and in 12 （12.5%）NPA samples from control children without wheezing， there being a statistically significant difference in the detection rates of HBOV between the symptomatic group and the asymptomatic group（χ2=7.377，P＜0.01）. Of 62 HBOV positive cases ， 28 patients （45.16%） mixed other viruses， in which 21 cases （33.87%） combined RSV infection. To compare the clinical characteristics between RSV single infection and HBOV single infection ， there was no statistical significance （P＞0.05）.No statistical significance was found beween HBOV single infection and HBOV mixed infection （P＞0.05）. HBOV viral load in HBOV-positive patients varied very broadly， from2.59 × 103 to5.36 × 109 copies/mL， the median viral load was 6.2×103 copies/mL（interquartile range，IQR 2.6×103 ~ 4.4×104 copies/mL）. A positive correlation between disease severity and the rank of viral load was found in cases with HBOV single infection（r = 0.752，P＜0.01），there being no correlation between disease severity and the rank of viral load found in cases with HBOV mixed infection（P＞0.05）.Conclusion　 Human bocavirus is prevalent in children with acute wheezing in Wenzhou area and is one of the major respiratory pathogens causing infants wheezing. The pathogenic role in HBOV single infection improve with the rise of viral load level.

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The significance of NSE and S-100β protein levels in CSF and serum of children with EV71 encephalitis. JIANG Qin，LIU Xing-li，ZENG Dong-sheng，LIN Xia，LI Jun.Qilu Children’s Hospital of 2012, 27(08): 612-614.  Abstract ( )   Objective    To study the early diagnostic significance of neuron-specific enolase（NSE）and S-100β protein in diagnosing EV71 encephalitis by detecting the level of NSE and S-100β protein in cerebrospinal fluid and serum in children infected with EV71. Methods    Totally 100 patients with EV71 encephalitis hospitalized in our hospital from May 2008 to December 2010 were chosen to be the study group，and 89 patients with other viral encephalitis and 22 patients without encephalitis hospitalized in our hospital in the same period were chosen to be the control group. The concentration of NSE and S-100β protein in cerebrospinal fluid and serum were detected by ELISA technique. Results    The concentration of NSE in cerebrospinal fluid，from patients in EV71 type encephalitis acute stage，was lower than other types of virus encephalitis group（P＜0.05）.In acute stage，the concentrations of NSE and S100β protein in cerebrospinal fluid and serum from patients with EV71 type encephalitis were obviously higher than EV71 ordinary type encephalitis（P＜0.05），and in convalescence stage，the concentrations were also higher than ordinary type（P＜0.01）.Conclusion    The concentrations of NSE and S-100β protein in cerebrospinal fluid and serum could be used to earlier evaluate the degree of injury and prognosis of intracranial infection in EV71 type encephalitis.

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Detection of human metapneumovirus in respiratory specimen by fluorescent quantitative PCR. WU Xu， WANG Tian-lin， CHEN Zhi-min， WU Yi-dong. Children’s Hospital，Zhejiang Uni 2012, 27(08): 615-618.  Abstract ( )   Objective    To establish fluorescent quantitative RT-PCR of detecting human metapneumovirus（HMPV），and to explore the HMPV infectious status and clinical features. Methods    Searching conservative HMPV N gene  segment for PCR amplification by DNAstar， compare and analyze whole genome sequence of series of HMPV； primers were designed and the method of RT-PCR to detect HMPV were established. HMPV in 623 clinical specimens from hospitalized children  from December 2009 to March 2010 were detected by fluorescent quantitative RT-PCR and direct immunofluorescence assay simultaneously. Results    Taking HMPV as template to carry out fluorescent quantitative PCR， we got positive result， but other common respiratory virus  and Mycoplasma pneumoniae showed negative results. By direct immunofluorescence assay， 10 cases were HMPV positive （1.61%）， and by fluorescent quantitative PCR method  28 cases were positive（4.49%）. Statistical significance between the two methods was found（χ2=16.05， P<0.01）. Of 10 cases with positive direct immunofluorescence assay， PCR all were positive， and 595 cases were negative by the two methods . A good correlation was found（r＝0.59，P<0.01）. The other 18 PCR-positive cases were negative by direct immunofluorescence method.The positive rate of HMPV in asthma was 15.4%（4/26）， much higher than 3.9% of the pneumonia group（20/515） and the tracheobronchitis group 0%（0/36），showing statistical significance（χ2＝11.22，P＝0.011）.The positive rate correlated with age，which in 0～1y group was 2.2%（9/404）， in＞1～3y group it was 13.1%（16/122）， in＞3～6y group it was 3.4%（2/58）， and  in larger-than-6 y group it was 2.6%（1/39）. The positive rate of HMPV in ＞1～3 y group was much higher than other groups（χ2＝26.44，P＝0.000）.Conclusion    By chosing conservative area of  HMPV N gene a pair of primers and single band of TaqMan probe are established， and the method of fluorescent quantitative RT-PCR is bulit up to detect HMPV. The sensitivity of fluorescent quantitative RT-PCR in detecting HMPV is much higher than that of direct immunofluorescence assay.  The positive rate of HMPV in asthma is obviously higher than that of pneumonia.  The positive rate of HMPV is the highest in children among ＞1～3 years old.

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Study of viral etiology of acute respiratory tract infection among pediatric outpatients in Beijing. LIU Chun-yan， ZHANG Hui， XIE Zheng-de， ZHAO Cheng-song. National Key Laborator 2012, 27(08): 619-621.  Abstract ( )    Objective    To investigate the viral pathogens among pediatric outpatients with acute respiratory tract infection in Beijing area. Methods    From March 2010 to February 2011， 540 pediatric outpatients with acute respiratory tract infection were involved in our study. One throat swab specimen was collected from each patient. Multiplex polymerase chain reactions were performed to detect respiratory viruses，including respiratory syncytial virus （RSV） type A and B， rhinovirus （RV）， influzenza virus （IFV） type A and B， parainfluenza virus （PIV） type 1-4， adenovirus （ADV）， enterovirus （EV）， human coronavirus （HCoV）， human metapneumonia virus （MPV） and human bocavirus （HBoV）. Results    At least one viral pathogen was detected in 306 out of 540 patients and the overall positive rate was 56.7%. Two or more viruses were identified in 34 cases and the mixed infection rate was 6.3%. IFV （16.7%） was the most prevalent virus， followed by ADV （12.8%）， EV （7.8%）， PIV （7.2%）， RSV （5.4%）， HMPV （5.0%）， RV （4.3%）， HCoV （3.0%） and HBoV infection （0.9%）. The most predominant type of IFV， PIV and RSV were IFVA， PIV3 and RSVA respectively. Conclusions    Viruses play an important role in respiratory tract infection in children. IFV， ADV， EV and PIV are the most important viral pathogens in pediatric outpatients with acute respiratory tract infection in Beijing area.