Epidemics of hand, foot and mouth disease (HFMD) have mainly been caused by Coxsackievirus A16 (CVA16) and Enterovirus A 71 (EV-A71), which circulated alternatively or together in the affected area. CVA16 has caused numerous outbreaks and epidemics in multiple countries and geographical regions, and has become an important public health problem. Based on an analysis of the complete VP1 coding region, all CVA16 strains can be divided into genotypes A, B1, and B2. Furthermore, genotype B1 can be divided into subgenotypes B1a, B1b, and B1c. After 2000, no reports of genotype B2 virus strains have been reported. All of the CVA16 strains reported in mainland China have belonged to subgenotypes B1a and B1b. Most CVA16-associated infections cause only mild symptoms; however, some CVA16 infections can lead to severe complications and even death. Vaccination is considered to be the most effective method to control the transmission and infection rate of this virus. A number of research groups are studying various vaccine types, including inactivated vaccines, genetic engineering vaccines, and DNA vaccines, amongst others. In this review, an overview is provided of the research advances in molecular epidemiology and vaccines of CVA16.
Animals ; China ; Coxsackievirus Infections ; epidemiology ; immunology ; prevention & control ; virology ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Humans ; Molecular Epidemiology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

Adult ; Coal Mining ; Dermatitis, Occupational ; epidemiology ; Humans ; Male ; Middle Aged ; Prevalence

Background Retinal angiomatous proliferation (RAP) is a subtype of neovascular age-related macular degeneration (AMD).There are currently very few studies on RAP.Objective This study was to explore the pathogenic mechanism of RAP in apolipoprotein E-deficient (ApoE-/-) mice with dyslipidemia.Methods Twenty-four 2-month-old SPF ApoE-/-mice were randomly divided into the high fat diet group and the normal diet group,and twelve 2-month-old C57BL/6 mice received the normal diet as controls.A diet with a higher content of fat was given for 4 consecutive months in the high fat diet group,and normal diet was given in the same way in the mice of the normal diet group.The mice were sacrificed at 6 months of age.The expression of vascular endothelial growth factor (VEGF) in the retinal pigment epithelial (RPE) cells,the expression of vascular endothelial growth factor receptor-2 (VEGFR-2) in the outer plexiform layer (OPL),microvascular density (MVD) and microvascular area (MVA) in the OPL were examined by immunohistochemistry and semi-quantitatively by histopathology with the Mias 2000 Imaging Analyzer System.The expression of VEGF protein in the retina was examined by Western blot.Results The MVD in the retinal OPL were (20.67±3.20) and (19.50± 1.87),respectively,in the ApoE-/-mice of the high fat diet group and the normal diet group,which were significantly higher than that (12.50±1.87) of the C57BL/6 normal diet group (all at P＜0.01).MVA in the retinal OPL were (626.49± 120.99) μm2 and (514.06±88.83) μm2 in the ApoE-/-mice of the high fat diet group and the normal diet group,respectively,showing a significant increase in comparison with the (336.52±84.96) μm2 of the C57BL/6 normal diet group (P＜0.01).The staining area of VEGF in RPE cells was (21 048±1849) μm2 in the ApoE-/-mice of the high fat diet group,showing a significant increase in comparison with the (17 116±2023) μm2 of the C57BL/6 normal diet group.However,no significant difference was found in the staining area of VEGF between the ApoE-/-mice with normal diet group and the C57BL/6 normal diet group ([17 854±2967] μm2 vs.[17 116±2023] μm2) (P＞0.05).Significant elevation was also seen in the staining area of VEGFR-2 in the retinal OPL of the ApoE-/-mice of the high fat diet group (12 193±3806)μm2 and the ApoE-/-mice of the normal diet group (11 969± 3616)xm2 compared with C57BL/6 mice of the normal diet group (5387±2225)μm2(all at P＜0.01).The relative expression values (VEGF/β-actin) of VEGF in the retinas were (1.51 ±0.32) and (1.17±0.39) in the ApoE-/-mice of the high fat diet group and the normal diet group,respectively,showing a significant increase in comparison with (0.28±0.14) of the C57BL/6 normal diet group (P＜0.01).Conclusions The expression of VEGF and VEGFR-2 in the retinas increases in the ApoE-/-mouse,which leads to the enlargement of MVD and MVA in the retinal OPL and subsequent RAP occurrence.

Objective This study was designed to explore the incidence rate of occuIt HBV infection in patients with anti-HBc positive alone and analyze the possible reasons of occuIt infection.Methods Sera of 183 patients carrying anti-HBc alone(A≤0.1) were collected and real-time PCR was used to select samples with HBV DNA positive.HBV pre-S/S amplification products were obtained by PCR,and clonal sequencing were then used for these samples with HBV DNA positive.Results DNA quantitative results of three samples were greater than 103 copies/ml in 183 samples,with a fraction of 1.6%.Pre-S/S sequencing results of two samples from these three samples were obtained.Point mutations within "a" determinant with Q129R/P mutations and co-existence of the mutant type and wild type were found in the two samples.Conclusions Occult HBV infection existed in samples with anti-HBc alone.Factors contributing to the loss of HBsAg detection by immunoassays include S gene mutations and low levels of cireulating antigen which are below the assay limit of detection.Occult HBV infection not only can lcad to a false clinical diagnosis,but also can result in hematological pollution due to such occult infection of blood donors.

Cognitive impairment is a common complication of diabetes. Hippocampus plays an important role in cognitive function. In hyperglycemia, synaptophysin, a crucial synaptic vesicle membrane protein in hippocampus neuron is found to be down-regulated. Recent evidences have shown that angiotensin IV can facilitate memory acquisition and recovery. However, whether it can also improve cognitive functions of diabetic rats with cognitive disorder, and the possible mechanisms are uncertain. Hence, the objectives of this study. Forty fi ve Sprague Dawley male rats were randomly divided into three groups: Control, diabetic group and diabetes with angiotensin IV treatment group. The cognitive functions, mainly learning and memory of the rats were evaluated using Morris water maze task. The synapses ultrastructure, relative mRNA concentrations and protein expression levels of synaptophysin in hippocampus CA1 area were estimated using transmission electron microscope, RT-PCR, immunohistochemistry and western blotting, respectively. Our study showed that in the diabetic rats with angiotensin IV treatment, the cognitive impairment as measured by Morris water maze task improved, the ultrastructure of synapses in hippocampus reversed, the relative mRNA concentrations and protein levels of synaptophysin in hippocampus signifi cantly increased, when compared with diabetic rats. We conclude that angiotensin IV plays an important role in improving cognitive function of diabetic rats. The possible mechanisms are up-regulating the expression of synaptophysin and normalizing the ultrastructure of synapses in hippocampus.

Objective To determine the therapeutic effect of recombined rat insulin-like growth factory 1 gene and transforming growth factor beta-1(TGF-?_1)gene on anterior cruciate ligament transection(ACLT)- induced osteoarthritis-like changes in NZW rabbit knee joints.Methods Eighteen NZW rabbits were divided into 3 groups randomly after osteoanhritis was established by ACLT and another six rabbits were used as normal control group(group 1).Chondrocytes which had been transfected with IGF-1 gene,co-transfected with TGF-?_1 and IGF-1 gene(group 3,4)were injected into the rabbits knee joints.Experimental control group(group 2)only had ACLT bul was not transfected.After 4,8 weeks,rabbits were sacrificed and their joints were evaluated by morphological grades,histological examination,in situ hybridization examination,immunohistochemistry exami- nation,and transmission electron microscopy examination(TEM).Results The morphological grades showed that the normal control group had a very significant difference with the experimental control group(P

To obtain a number of penicillinases and degrade penicillin in milk by using the penicillinases,the gene encoding penicillinase was amplified by PCR from Bacillus cereus ATCC10987,cloned into pET28a(+) ,transformed into E. coli BL21;analysis of SDS-PAGE and penicillinase activity of the recombinant protein were done under induction of IPTG and the result showed that the maximum penicillinase activity reached 480 U/mL;the purity of penicillinase purified by Ni2+ Purification System was more than 90%;the immobilized penicillinases were obtained by sodium periodate method and the residual quantity of penicillin in milk(containing 0.5 U penicillin G/mL) was less than 4 ppb after degraded by the immobilized penicillinase.

Objective To evaluate the significance of human interleukin-31(IL-31)in the pathogenesis of atopic dermatitis and its correlation with pruritus in patients with atopic dermatitis(AD).Methods Twenty-two children with mild to severe atopic dermatitis and 22 age-matched healthy controls were included in this study.Patients and controls were randomly and equally assigned into stimulation and non-stimulation groups.Venous blood samples were obtained from all participants,peripheral blood mononuclear cells were isolated from these samples and cultured with(stimulation groups)or without(non-stimulation groups)staphylococcal enterotoxin B(SEB)for 24 hours.Then,the mRNA expression of IL-31 on PBMCs was assessed via real-time reverse transcription-PCR.ELISA was used to detect the total serum IgE level in these objects.The severity of AD in patients was rated according to scoring atopic dermatitis(SCORAD).The relationship between the mRNA expression of IL-31 and the level of serum total IgE.severity of atopic dermatitis,and degree of pruritus.was evaluated.Results The expression of IL-31 mRNA on non-stimulated PBMCs from patients was 23.2 folds as high as that from the healthy controls(P＜0.01).The stimulation with SEB upregulated the mRNA expression of IL-31 on PBMCs.and the increase on PBMCs from patients was 20.44 times of that from the controls.The total serum IgE level was 260.05 IU/mL(5.9-1131.01 IU/mL)and 17.7 IU/mL(5-140.7 IU/mL)in the Patients and controls respectively(P＜0.01).There was no significant correlation between the mRNA expression of IL-31 and disease severity or total serum IgE level(r=0.07.0.22respectively.both P＞0.05)in patients witll AD.Condusions IL.3 1 is involved in t11e pathogenesis of AD,which is unlikely to be IgE-dependent.SEB can induce the rapid expression of IL-31 on PBMCs of healthy human,and is an important modulator for the production of IL-31.

Objective To evaluate the capacity of atopy patch test in diagnosis of food allergy in children with atopic dermatitis (AD).Methods Egg and milk,as the most common food allergens among Chinese children,were employed in this study.Skin prick test (SPT) and atopy patch test (APT) with fresh egg and milk were carried out in 68 children with AD.Oral food challenges in an open style were performed to confirm the diagnosis of food allergy.Resuits Of these patients,58(85.3%)were sensitive to egg,40(58.8%)to milk and 34(50.0%) to both.Of 98 patients with positive challenge,47 showed late response,10 immediate reactions.and 41 mixed reactions.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and the agreement with food challenges in diagnosis of egg/milk allergy were 96.6%/67.5%.90.0%/82.1%,98.2%/84.4%,81.8%/63.9% and 95.6%/73.5%,respectively for APT alone,37.9%/30.0%,100%/89.3%,100%/80.0%,21.7%/47.2% and 47.1%/54.4%,respectively for SPT alone.APT was found to be more sensitive in diagnosis of late-phase reactions than SPT (P<0.01).No significant difference was found in the sensitivity between APT alone and the combination of APT and SPT in parallel algorithm for diagnosis of egg or milk allergy (x2=0.509,0.549,both P>0.05) or in the specificity between APT alone and that in serial algorithm( P=1.000;x2=3.514,P>0.05).Conclusions APT is superior to SPT in diagnosis of late responses to food,and the combination of SPT and APT does not facilitate the diagnosis of food or milk allergy compared with APT alone.

This study aims to construct inactivated coxsackievirus A16 (CVA16) vaccine and to investigate its protective effect in ICR mice. A clinical isolate of CVA16, 521-01T, was cultured in VERO cells, inactivated by formaldehyde, and purified by ultracentrifugation for vaccine preparation. Purity and other characteristics of the vaccine were determined by SDS-PAGE and Western blot. Female ICR mice were subcutaneously inoculated with inactivated CVA16 or Al(OH)3-absorbed CVA16, followed by booster immunization at the end of 2 and 4 weeks. CVA16-specific IgG titers in serum were determined by ELISA, and titers of neutralizing antibodies were determined by viral neutralization assay. The immunity of T lymphocytes was evaluated by IFN-gamma ELISPOT assay. The protective effect was evaluated by challenging the neonatal offspring (< 48 hours) of vaccinated female mice with 1 000 LD50 of CVA16 521-01T. The mortality rates of different groups were compared. The results showed that Al(OH)3 +CVA16 could induce high titers of specific IgG antibodies in ICR mice. After being boosted two times, the serum IgG antibody titer could reach up to 1 : 1 x 10(5) (P = 0.000), and neutralizing antibody titer was higher than 1 : 256. Additionally, more spot forming cells were induced in the immunized groups than in the negative controls. The maternal antibodies showed protective effect in 100% of the neonatal mice challenged with 1 000 LD50 of CVA16 521-01T. The inactivated CVA16 vaccine has ideal immunogenicity and immunoprotective effect. This research lays a foundation for the development and evaluation of CVA16 vaccines.
Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Enterovirus ; immunology ; Enterovirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Immunization ; Mice ; Mice, Inbred ICR ; T-Lymphocytes ; immunology ; virology ; Vaccines, Inactivated ; administration & dosage ; immunology ; Viral Vaccines ; administration & dosage ; immunology