| Purpose:EV71 infection has a serious impact on children health,especially EV71-related encephalitis,which is caused by the neurotropic,positive-sense RNA virus,enterovirus71.Since EV71 was first isolated in California in 1969,many outbreaks with fetal cases have been reported.However,not all of the patients were severely infected,and the study found that the host gene was associated with the outcome of the disease.Toll-like 3(TLR3),as a receptor of virus ds-RNA,plays a key role in EV71 infection.Organism will produce some pathogen associated molecular patterns(PAMPs)when pathogens infect host,which can be distinguished by pathogen-recognition receptors(PRRs),and lead to cell signal reaction like typeⅠIFNs/IL10 pathway.The anti-inflammatory master regulators,typeⅠIFNs and IL-10 are critical to protect the host from tissue damage during acute phases of immune responses.A possible association between EV71 infection in a Chinese population and the single nucleotide polymorphism(rs5743303)in promoter of theTLR3 gene,was investigated.At the same time,we detected whether there was a relationship between the level of IFN /IL-10 which are the downstream cytokines of TLR3 and the polymorphism of the gene.We should understand the relationship between host immunity and the prognosis of the disease from the gene level,and provide a molecular basis for the diagnosis,treatment and immune prevention of the disease.Methods:The genotypic and allelic frequencies of these polymorphisms were analyzed in 221EV71 infection patients,including mild cases(153)and severe cases(68),as well as in a control population(175 randomly selected healthy children).EV71 virus nucleic acid is positive in children with throat swab,feces,cerebrospinal fluid through RT-PCR detection.The improved multiplex ligation detection reaction(iMLDR)method was used to detect the TLR3 rs5743303 polymorphism,which is located in promoter of TLR3 on chromosome 4q35.The Interferon α/β and IL-10 concentration in serum or CSF was determined using a commercial enzyme-linked immune sorbent assay kit(R&D System,Minneapolis,MN)according to the manufacturer ’s instructions.Complate clinical data collection and statistics of patients with duration of fever,white blood cells,CRP,ALT,AST and myocardial enzymes.All statistical analysis was carried out using the Statistical Package for Social Science(IBM SPSS software,USA),version16.0,and a P-value less than 0.05 was considered significant.Results:1.There was no significant difference in the frequency of genotype and allele frequency between the control group and the EV71 infection group.The frequency of A allele and AA genotype in severe EV71 patients was significantly higher than that in the control group(P = 0.004 and P = 0.009).The frequency of A allele and AA genotype of severe EV71 infection was significantly higher than that of children with mild infection(p=0.015,P=0.025).2.The level of serum IFN-α(40.76 + 3.78 pg/mL)in the EV 71 infection group was significantly higher than that in the healthy control group(0.20 + 0.12 pg/mL,P=0.000).The level of IFN-α(45.64 + 3.18pg/m L)in the serum of EV71 infected patients with AT+TT genotype was significantly higher than that of the AA genotype(9.34 + 1.24pg/ml,P=0.026).The level of IFN-β in serum of EV 71 infected people was not significantly different from that of the healthy control group(P>0.05).3.The level of serum IL-10(28.82 + 1.30 pg/mL)in the EV 71 infection group was significantly higher than that in the control group(0.35 + 0.14 pg/mL,P=0.000).In patients with EV71 infection,the serum IL-10 level of AT+TT genotype patients(39.31 +3.58 pg / m L)was significantly higher than that of AA genotype patients(14.14 +6.64pg/ml)(P=0.002).4.A carriers(AT + AA)(6.2mmol/l)had significantly higher blood BG levels compared to TT carriers(5.1mmol/l)in patients with EV71 encephalitis(p<0.05).We didn’t find the difference about CRP,duration of fever,ALT,AST and so on.Conclusions:1.Our results indicate that the relationship between TLR3 gene variant and severe EV71 infection is more closer,contrary to healthy children.It could be a genetic risk factor for the development of severe EV71.2.We suggest that this polymorphism conferred reducing secretion of cytokines with antiviral activity,such as IFNα/βand IL-10 secretion.Purpose: Cohen syndrome is a rare,genetic,connective-tissue disorder,which is caused by mutations in the gene COH1(VPS13B)at the chromosome 8q22.The disease is rare reported,which major clinical features include postnatal microcephaly,obesity,short stature,intellectual disability,progressive retinal dystrophy,intermittent neutropenia and many other unusual facial feature.We report three patients in China who were diagnosed with Cohen syndrome by genetic testing and clinical manifestations.At the same time,we review the related literature,and further expound the molecular mechanism of the disease,a variety of clinical manifestations,treatment and prognosis.Methods: We collected 2ml vein blood from each patient and from each inherited parent of the subject separately with the signed informed consent.Then the blood samples were sent to Beijing Kangso Medical Inspection for gene test of intelligence and motor developmental delay diseases,including VSP13 B gene testing.The next generation sequencing was used to complete the genome sequencing of children.The first generation sequencing was used to validate the related mutation sites of parents.Then complete prediction of mutant gene related proteins.And the clinical data of the patients were collected for analysis.Results: These 3 patients were found mutations in the VPS13 B,c.3203C>T,8016+7G>C;c.2199C>A,c.553 T>C;c.6940 + 1 G > T,c.9852-9855 del.And protein prediction has pathogenicity.Their VPS13 B gene mutation was a pathogenic gene mutation in combination with the clinical manifestation and the parents’ corresponding gene mutation sites.Conclusions:1.The clinical manifestations of Cohen syndrome are complex and varied.We should pay attention to the detailed analysis of the patient’s clinical data and carry out a more accurate gene detection.2.The VPS13 B gene mutation in Cohen syndrome is varied,and the mutation locations of the 3 patients have enriched the mutant gene pool.3.Early diagnosis of Cohen syndrome and early intervention can greatly improve the quality of the patients’ life and be conducive to social stability. |
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