Association of interleukin-10 gene polymorphism with enterovirus 71 infection in children / 中国当代儿科杂志

OBJECTIVE@#To study the association of interleukin-10 (IL-10) -1082A/G, -819C/T, and -592C/A polymorphisms with IL-10 level and the severity of enterovirus 71 (EV71) infection in children.@*METHODS@#A total of 137 children with hand-foot-mouth disease due to EV71 infection were enrolled as EV71 infection group, which was further divided into mild group with 91 children and severe group with 46 children, and 122 healthy children who underwent physical examination were enrolled as healthy control group. Related clinical data were collected. ELISA was used to measure the serum level of IL-10, and polymerase chain reaction-restriction fragment length polymorphism was used to analyze IL-10 -1082A/G, -819C/T and -592C/A polymorphisms.@*RESULTS@#Compared with the healthy control group, the children with EV71 infection had significantly higher frequency of -1082 AA genotype and A allele (P0.05). The severe group had a significantly higher serum level of IL-10 than the mild group and the healthy control group. IL-10 -1082 AA genotype, -819 TT genotype, and -592 AA genotype were associated with the low expression of IL-10 (P0.05).@*CONCLUSIONS@#IL-10 gene polymorphisms are associated with IL-10 expression and the severity of EV71 infection in children.

Polymorphism of OAS2 rs739901 C/A Involves the Susceptibility to EV71 Infection in Chinese Children / 华中科技大学学报（医学）（英德文版）

This study aimed to assess the relationship of OAS2 rs739901 5'-flanking C/A polymorphisms with the susceptibility to Enterovirus-71 (EV71) infection.We investigated 294 hand-foot-mouth disease (HFMD) Chinese children with EV71 infection (165 mild cases and 129 encephalitis cases).The improved multiplex ligation detection reaction (iMLDR) technique was used to test the genotypes.In EV71-infected patients,the CA genotype distribution (P=0.007),A allele frequency (OR 1.32,95％ CI 1.0-1.7,P=0.034)and CA+AA carriage frequency (P=0.003) of OAS2 rs739901 5'-flanking were obviously elevated as compared with controls,but there were no statistically significant differences between mild cases and encephalitis cases.In EV71-infected patients,the counts of white blood cells (P=0.034) and blood glucose concentrations (P=0.042) were raised in A carriers (CA+AA).Among different genotypes of encephalitis cases,the contents of cerebrospinal fluid (CSF) showed no significant differences.IFN-γ levels in EV71-infected patients were higher than those in controls (mild group vs.control group,P＜0.01;encephalitis group vs.control group,P＜0.001).In encephalitis cases,IFN-γ levels were reduced (P＜0.05) in A carriers compared to CC genotype,however,there were no significant differences between genotypes CA and AA (P=0.226).These findings suggest that OAS2 rs739901 5'-flanking C/A genetic polymorphisms involve the susceptibility to EV71 infection,and A allele might be a risk factor of the susceptibility to EV-71 infection.

Polymorphism of OAS2 rs739901 C/A Involves the Susceptibility to EV71 Infection in Chinese Children / 华中科技大学学报（医学）（英德文版）

This study aimed to assess the relationship of OAS2 rs739901 5'-flanking C/A polymorphisms with the susceptibility to Enterovirus-71 (EV71) infection.We investigated 294 hand-foot-mouth disease (HFMD) Chinese children with EV71 infection (165 mild cases and 129 encephalitis cases).The improved multiplex ligation detection reaction (iMLDR) technique was used to test the genotypes.In EV71-infected patients,the CA genotype distribution (P=0.007),A allele frequency (OR 1.32,95％ CI 1.0-1.7,P=0.034)and CA+AA carriage frequency (P=0.003) of OAS2 rs739901 5'-flanking were obviously elevated as compared with controls,but there were no statistically significant differences between mild cases and encephalitis cases.In EV71-infected patients,the counts of white blood cells (P=0.034) and blood glucose concentrations (P=0.042) were raised in A carriers (CA+AA).Among different genotypes of encephalitis cases,the contents of cerebrospinal fluid (CSF) showed no significant differences.IFN-γ levels in EV71-infected patients were higher than those in controls (mild group vs.control group,P＜0.01;encephalitis group vs.control group,P＜0.001).In encephalitis cases,IFN-γ levels were reduced (P＜0.05) in A carriers compared to CC genotype,however,there were no significant differences between genotypes CA and AA (P=0.226).These findings suggest that OAS2 rs739901 5'-flanking C/A genetic polymorphisms involve the susceptibility to EV71 infection,and A allele might be a risk factor of the susceptibility to EV-71 infection.

Association of TLR3-1377C/T gene polymorphisms and expression with susceptibility to enterovirus 71 encephalitis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the association of gene polymorphisms of Toll-like receptor 3 (TLR3)-1377C/T and expression of TLR3 with the susceptibility to enterovirus 71 (EV71) encephalitis in children.</p><p><b>METHODS</b>A total of 187 children with EV71 infection (59 children in the encephalitis group and 128 in the non-encephalitis group) and 232 children who underwent physical examination were enrolled in the case-control study. Polymerase chain reaction-restriction fragment length polymorphism was used to detect the TLR3-1377C/T gene polymorphisms. ELISA was used to measure the serum level of TLR3.</p><p><b>RESULTS</b>There were no significant differences in the genotype and allele frequencies of TLR3-1377C/T between the non-encephalitis group and the encephalitis group. Compared with the control group, the encephalitis group and the non-encephalitis group had significant increases in the serum level of TLR3 (P<0.05), and the non-encephalitis group had the highest level (P<0.05). The encephalitis group had a significantly higher EV71 viral load than the non-encephalitis group (P<0.01). The children aged <1 year or ≥1 year in the encephalitis group and the non-encephalitis group had significant increases in the serum level of TLR3 compared with their counterparts in the control group (P<0.05), and the children aged <1 year or ≥1 year in the non-encephalitis group had a significantly higher serum level of TLR3 than those in the encephalitis group (P<0.05). In the encephalitis group, the children aged ≥1 year had a significantly higher TLR3 concentration than those aged <1 year (P<0.05), and there were no significant differences in the TLR3 concentration between the children aged ≥1 year and <1 year in the non-encephalitis group and the control group. In the encephalitis group, the proportion of children aged <1 year was significantly higher than those aged ≥1 year (P<0.05).</p><p><b>CONCLUSIONS</b>The TLR3-1377C/T gene polymorphisms are not significantly associated with the development of EV71 encephalitis. Low expression of TLR3 might weaken the inhibitory effect on virus replication and promote the development of EV71 encephalitis. The deficiency in the expression of TLR3 in serum after EV71 infection might be an important factor for the development of encephalitis in infants.</p>

Relationship between variation of coxsackievirus B3 VP1 sequence from cerebrospinal fluid of children and severity of damage to central nervous system / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the possible relationship between variation of coxsackievirus B3 (CoxB3) VP1 sequence from cerebrospinal fluid of children with severe and mild central nervous system (CNS) infection and damage to CNS in children from Shandong province.</p><p><b>METHODS</b>The enteroviruses were detected using VP1 typing and sequencing primer for enteroviruses from 73 enterovirus-infected cases confirmed by detection of cerebrospinal fluid by enteroviruses common primer. VP1 sequences (450 nucleotides) were determined and analyzed for 21 CoxB3 enteroviruses strains isolated in Qingdao and Binzhou, and were compared with that of BLAST search procedures from GeneBank in NCBI. The variation of VP1 gene and amino acids sequence of CoxB3 enteroviruses was analyzed for severe and mild CNS infection.</p><p><b>RESULTS</b>The nucleotide homogeneity of these CoxB3 appeared to be 97% - 99%, however, the homogeneity among different genotypes were 83% - 76%. Replacement of glutamine by histidine at amino acid locus 856 of VP1 CoxB3 was found in 4 cases with severe encephalitis. There were different variation in VP1 nucleotide sequence of CoxB3 in 3 cases with mild encephalitis and 14 cases with meningitis, but amino acids sequences had no regular variation. The modified Glasgow's coma score was below 7 in all the 4 cases with severe encephalitis. Of these 4 cases, 3 had consciousness disturbance for less than 3 days. Lethargy, restlessness and psychiatric symptoms were major manifestations, of whom 3 also had dysphagia, 1 had encephalatrophy obviously, Glasgow's coma score was 3, deep coma lasted for 9 days, and had concomitant fatal epileptic attacks. Of these 4 cases, 2 completely recovered, 1 had high muscle tone, 1 remained under anti-epileptic drug treatment at follow-up 6 months later.</p><p><b>CONCLUSION</b>There were a small epidemic of CoxB3 CNS infection in children in 2005 in this area. The amino acid variation of CoxB3 VP1 possibly caused increased viral virulence and caused damage to CNS.</p>

Prevention of beta cell dysfunction and apoptosis by adenoviral gene transfer of rat insulin-like growth factor 1 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Islet beta-cells are almost completely destroyed when patients with type 1 diabete are diagnosed. To date, insulin substitute therapy is still one of the main treatments. The cure of type 1 diabetes requires beta-cell regeneration from islet cell precursors and prevention of recurring autoimmunity. Therefore, beta-cell regeneration and proliferation emerge as a new research focus on therapy for type 1 diabetes. Islet beta-cell regeneration and development are controlled by many growth factors, especially insulin-like growth factor-1 (IGF-1).</p><p><b>METHODS</b>Recombinant adenovirus encoding rat IGF-1 (rIGF-1) was constructed and transduced into rat beta-cells, RINm5F cells. Western blotting analysis and ELISA were used to detect rIGF-1 protein. Streptozotocin (STZ) was used to induce RINm5F cell destruction. The level of nitric oxide (NO) was detected in cell culture supernatants by the Griess reaction. Islet cell function was evaluated by glucose-stimulated insulin production. Flow cytometry analysis was further used to investigate the apoptosis of RINm5F cells. Thiaoollyl blue viability assay was applied to determine cell viability.</p><p><b>RESULTS</b>The recombined adenovirus-rIGF-1 was successfully constructed and the titer was 4.0 x 10(8) pfu/ml. The rIGF-1 protein was effectively expressed in the RINm5F cells and cell culture supernatants. rIGF-1 expression remarkably inhibited STZ-induced islet cell apoptosis and significantly decreased the level of NO. Furthermore, IGF-1 expression also significantly protected insulin secretion and cell proliferation in a time-dependent manner.</p><p><b>CONCLUSIONS</b>Our study suggests that locally produced rIGF-I from RINm5F cells may be beneficial in maintaining beta-cell function, protecting beta-cells from the destruction of apoptosis factors and promoting beta-cell survival and proliferation. IGF-I might be considered as a candidate gene in gene therapy for type 1 diabetes. In addition, it appears that the apoptosis induced by STZ may be NO-dependent.</p>

Effects of astragalus membranaceus on TH cell subset function in children with recurrent tonsillitis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To observe the TH cell subset function in children with recurrent tonsillitis (RT) at the remission stage and to study the effects of astragalus membranacus (AM) on TH cell subset function.</p><p><b>METHODS</b>The peripheral blood mononuclear cells (PBMC) from 27 children with RT at the remission stage were stimulated with either phytohemagalutinin (PHA) (RT-PHA group) or PHA together with AM (RT-AM group) and were then cultured in vitro for 48 hrs. The samples from 21 healthy children stimulated with PHA were used as the Control group. The levels of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) in the supernatants of PBMC were detected using ELISA.</p><p><b>RESULTS</b>The IFN-gamma level and the ratio of IFN-gamma/IL-4 in the RT-PHA group were statistically lower than those in the Control group (P < 0.01). The level of IFN-gamma and the ratio of IFN-gamma/IL-4 in the RT-AM group were markedly higher than those in the RT-PHA group (P < 0.01), but were significantly lower than those in the Control group (P < 0.05). There were no differences in the IL-4 level among the three groups.</p><p><b>CONCLUSIONS</b>TH1 cell subset dysfunction may exit in RT children at the remission stage, suggesting that TH1 cell subset dysfunction plays an important role in the pathogenesis of RT. AM can improve TH1 cell subset function and therefore shows an important significance in treating RT.</p>

Changes of Activated T Cell Disfunction in Children with Refractory Mycoplasma Pneumoniae Pneumonia / 实用儿科临床杂志

Objective To study disfunction of activated T cell from peripheral blood and explore its role on pathogenesis in patients with refractory mycoplasma pneumoniae pneumonia (MPP).Methods T cell subsets and expression of membrane molecule on activated T cell in peripheral blood were measured in 26 refractory MPP patients by using immunofluorescence and flowcytometry,and compared with those of 18 control healthy children.Results The percentage of CD4~+ cell from peripheral blood in refractory MPP children was significantly lower than that in control group(P0.05).Conclusions There are disfunctions of T cell in refractory MPP patients in acute period,and the abnormal T cell activated disfunction may play an important role on pathogenesis.

Regulatory effect of zengmian mixture on T-lymphocyte dysfunction in children with repeated lower respiratory tract infection of both Qi-Yin deficiency type / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the T-lymphocyte dysfunction in children with repeated infection of lower respiratory tract of both Qi-Yin deficiency type (RIR-QYD) and the immune regulatory effect of zengmian mixture (ZMM), to provide theoretical basis for the effective therapy.</p><p><b>METHODS</b>Peripheral T-lymphocyte subsets and expressions of T-lymphocyte activating related surface molecules (CD3+/HLA-DR+ and CD3+/CD25+, etc.) in children with RIR-QYD, 31 of mild type and 28 of severe type cases, were investigated before administration of ZMM and after treatment of ZMM for 3-6 months (non-infectious stage), using immune fluorescent labelling and flow cytometric technique.</p><p><b>RESULTS</b>In the patients with mild RIR-QYD, the expression rate of CD4+ and CD3+/HLA-DR+ activated T-cells before treatment were all obviously lowered, after 3 months treatment, the positive rate of CD4+, CD3+/HLA-DR- resting T-cell, CD3+/HLA-DR+ activated T-cell and CD3+/CD25+ express IL-2R T-cells were all obviously lowered, but after treatment for 6 months, only that of CD3+/HLA-DR+ activated T-cells was lower than that in the control group. In the patients with severe RRI-QYD before treatment, the expression rate of CD3+, CD4+, CD3+/HLA-DR-, CD3+/HLA-DR+ and CD3+/CD25+ all lowered, while after 3-6 months treatment, some recoveries were shown in these parameters but still lower than those in the control group. The total effective rate of ZMM for mild patients was 100%, and the markedly effective rate 78.9%, while for severe cases, the total effective rate was 90.9% and the markedly effective rate 68.2%.</p><p><b>CONCLUSION</b>In patients with RIR-QYD, the T-cells decreased with activating dysfunction, the severity of disease is in accordance with the degree of T-cell activating dysfunction. ZMM shows markedly clinical effect in treating RIR-QYD and evident regulatory effect on T-cell dysfunction, but a long-term treatment is needed for the recovery of laboratory parameters.</p>

Analysis of 187 children with enteroviral central nervous system infection in Shandong area / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the diagnostic potential of previously published enterovirus (EV) reverse transcription polymerase chain reaction (RT-PCR) assay in detection of EV in CSF samples from children with a diagnosis of aseptic meningitis and to investigate the clinical characteristics of the patients seen in Shandong.</p><p><b>METHODS</b>EV RNA was detected in 187 CSF samples and serum and/or urine samples of a part of patients by RT-PCR and viral culture technique.</p><p><b>RESULTS</b>RT-PCR was positive in all 62 CSF specimens which were positive by cell culture (100%). In addition, 93 of 125 (74.4%) CSF samples negative by cell culture were RT-PCR positive. In 4 of these 93 (4.3%) patients, viral culture of specimens from other sites (serum or urine) was also positive. The sensitivity of CSF RT-PCR based on clinical diagnosis in patients with meningitis of negative bacterial culture results was 82.9% (155/187), which was considerably higher than the sensitivity of CSF virus culture 33.2% (62/187). The results of RT-PCR can be reported within 4 hours, whereas the viral culture of CSF requires 4.6 days for a cytopathic effect to develop. EV meningitis occurred in a sporadic form and in some areas there were outbreaks. The clinical characteristics of 155 patients with EV meningitis were different in different age groups.</p><p><b>CONCLUSION</b>EV was one of the most common causes of aseptic meningitis in Shandong area. The RT-PCR assay was rapid, sensitive and specific for the diagnosis of EV meningitis and may be a potential tests to shorten hospital stay and reduce the use of antibiotics.</p>

Study on T-Cell and T-Helper Cell Subpopulation Function in Children with Recurrent Tonsillitis in Remission Period / 实用儿科临床杂志

Objective To study the state of T-cell and T-helper cell subpopulation in children with recurrent tonsillitis(RT) in remission period and make clear its role on pathogenesis in children with RT.Methods The expression of membrane molecule on T-cell subsets was analyzed in 27 RT children and 21 control health children by using immunofluorescence and flowcytometry,the levels of interferon-?(IFN-?) and interleukin-4(IL-4) in culture supernatant of peripheral blood mononuclear cells(PBMC) were detected by enzyme linked immunoadsorbent assay(ELISA).Results The CD4~+ and CD3~+ cell were significantly lower in RT children than those of the controls(P0.05),a ratio of CD4~+/CD8~+ was disproportion(P0.05),the ratio of Th1/Th2 was disproportion(P