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

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

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.

OBJECTIVETo observe morphological changes of enteric nervous system (ENS)-interstitial cells of Cajal (ICC)-smooth muscle cell (SMC) structure injury in deep muscle nerve plexus offunctional dyspepsia (FD) rats, and the repair of Shuwei Decoction (SD) on it, and to explore its effecton FD.

METHODSTotally 72 rats were randomly divided into the control group, the model group, the lowdose SD group, the medium dose SD group, and the high dose SD group, the Mosapride group, 12 ineach group. Rats in the low dose SD group, the medium dose SD group, and the high dose SD group were intragastrically fed with SD at 0.767, 1.534, 3.068 g/mL, respectively. Rats in the Mosapride group were intragastrically fed with Mosapride (1.37 mg/kg). FD rat model with Gan depression Pi deficiency syndrome (GDPDS) was established using complex pathogenic factors. Corresponding liquors were respectively administered to rats in corresponding groups from the 3rd day after modeling. Distilled water(10 mL/kg) was administered to rats in the control group and the model group, once per day for 14 successive days. Rats were sacrificed and small intestine tissues collected for observing ENS-ICC-SMC structure injury using immunofluorescence double labeling, laser scanning confocal microscope, and transmission electron microscope at day 15. Repair of SD on it was also observed.

RESULTSENS-ICC SMC structure was incomplete, with obvious injury in mutual link of ICC, ICC, SMC, and connecting structure. ENS-ICC-SMC structure was more complete in high, medium, and low dose SD groups, with close link of ICC and SMO. Their connecting structures were in good conditions.

CONCLUSIONSD could keep the integrity of ENS-ICC-SMC structure by promoting regeneration and morphology of ICC, thereby, improving gastrointestinal movement disorder and showing therapeutic effect on FD.

Animals ; Benzamides ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Dyspepsia ; drug therapy ; Enteric Nervous System ; drug effects ; Interstitial Cells of Cajal ; drug effects ; Morpholines ; pharmacology ; Muscle, Smooth ; drug effects ; Random Allocation ; Rats

OBJECTIVE: To investigate the clinical value of negative pressure tympanograms for diagnosis of middle ear effusion in adults. METHOD: GSI-Tympstar II Middle Ear Analyzer of America was used to test tympanogram peak pressure (TPP); tympanogram width (TW); middle ear resonant frequency and stapedius muscle reflex for sceptical secretory otitis media (SOM) adult patients. Patients (207 ears) with C type tympanograms accepted tympanocentesis after tests. According to middle ear effusion quantity, 207 ears have been divided to three groups: non-effusion group (123 ears), small-effusion group (45 ears) and mickle-effusion group (39 ears). Acoustic immittance and ipsilateral stapedius muscle reflex were statistically analysed. RESULT: About TPP and resonant frequency, non-effusion group and small-effusion group have statistical difference (P < 0.05); non-effusion group and mickle-effusion group have significant statistics different (P < 0.01). About TW, non-effusion group have significant difference from mickle-effusion group (P < 0.01). There was statistical difference on ipsilateral stapedius muscle reflex (P < 0.01) among three groups. CONCLUSION Negative pressure tympanograms has diagnostic value for middle ear effusion combined to TPP, TW, middle ear resonant frequency and ipsilateral stapedius muscle reflex.
Acoustic Impedance Tests ; methods ; Adolescent ; Adult ; Aged ; Ear, Middle ; pathology ; Female ; Humans ; Male ; Middle Aged ; Otitis Media with Effusion ; diagnosis ; Young Adult

Objective:To investigate the clinical significance of bone metabolism indexes and human leukocyte antigen B27 (HLA-B27)in ankylosing spondylitis(AS)and analyze their diagnostic value. Methods:The subjects were 94 cases of AS patients,239 cases with other diseases and 80 healthy controls, and the results were retrospectively surveyed. Results:In the AS group, the concentration of the β-collagen specific sequence (β-CTX ) was higher ( P<0. 05 ) while the concentrations of osteocalcin ( OC ) , 25-hydroxyvitamin D[25-(OH)D] and parathyroid hormone(PTH)were lower than those in the control group. In the AS group,there was a positive correlation between the concentration of C-reactive protein(CRP)and β-CTX(P<0. 01),and the concentration of CRP was negatively correlated with the concentration of 25-(OH)D(P<0. 05). However,the concentrations of other bone metabolic indexes had no correlation with the concentration of CRP(P<0. 05). The positive rate of HLA-B27 in the AS group was significantly higher than that in other groups(P<0. 05),and HLA-B27 had highly sensitive and specific in the diagnosis of AS. In the AS group,the sensitivity of the concentration of 25-( OH) D was higher than β-CTX,while its specificity was lower than β-CTX. Conclusion:Bone metabolic indexes have great value in early screening and clinical diagnosis of AS,especially β-CTX and 25-( OH) D were more obvious.

To analyze the epidemiological and etiological characteristics of Hand-Foot-and-Mouth disease (HFMD) in Shanghai in 2009, epidemiological data was retrieved from the National Notifiable Disease Report System (NNDRS). Nucleic acid of enterovirus (EV) was detected by real-time RT-PCR from 799 HFMD cases from 15 districts/counties in Shanghai; the complete sequences of VP1 encoding region of several identified EV71 strains and sequences of VP4 encoding region of several untyped EV were determined and analyzed. Analysis and summary of the epidemiological data was conducted with Microsoft Excel, and sequence analyses were conducted with both BioEdit and MEGA software. Untyped EV was identified through comparing the VP4 sequence to sequence database using BLAST online service. It was showed that all the 18 districts/counties had reported HFMD cases; children less than 6 years old were the most susceptible population group; the peak of epidemics of HFMD was from April to July; EV71 and Coxsackievirus A16 (CA16) were the major pathogens for this epidemic, but the constituent ratio of EV71 and CA16 was different in different months and regions; CA16 infection was mainly responsible for the mild HFMD, but EV71 for most of the severe cases; EV71 strains of Shanghai were clustered with representatives of subgenotype C4a and showed the highest identity to them, based on the sequence analyses of VP1 encoding region; 2 of the untyped EV were identified as CA2 and CA10 respectively. All the results indicated that EV71 and CA16 were the major pathogens for the epidemic of HFMD in Shanghai, 2009; the circulating EV71 belonged to subgenotype C4a. Besides, other types of EV (for example: CA2 and CA10) were also responsible for a few of the HFMD cases.
Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Epidemics ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny

China ; epidemiology ; Enterovirus ; genetics ; isolation & purification ; Genes, Viral ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans

OBJECTIVETo study the effect of astragaloside IV on the expression of cytokines in bone mesenchymal stem cells (MSCs) in rats.

METHODSMSCs were isolated from Wistar rats by the method of adhesive cultiration and clone, and then their biological activities were assessed using indirect immunofluorescence. Proliferation of MSCs stimulated with astragaloside IV was ascertained by the MTT method. Expression of cytokines was ascertained using RT-PCR in MSCs with astragaloside IV stimulation or not.

RESULTSMSCs were effectively isolated and purified in vitro, and had expression of many cytokines except IL-3, such as stem cell factor (SCF), thrombopoietin (TPO), granulocyte macrophage colony stimulating factor (GM-CSF) and transforming growth factor (TGF-beta1). Astragaloside IV stimulation promoted MSCs proliferation, and 200 mg/mL astragaloside IV treatment produced a peak effect 72 hrs after culture. The SCF expression in MSCs stimulated with astragaloside IV increased significantly compared with that in MSCs without astragaloside IV stimulation.

CONCLUSIONSAstragaloside IV may promote MSCs proliferation and increase SCF secretion in vitro.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Proliferation ; drug effects ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; secretion ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Saponins ; pharmacology ; Stem Cell Factor ; biosynthesis ; genetics ; secretion ; Triterpenes ; pharmacology

OBJECTIVETo investigate short-stage results of audiological change of nasopharyngeal carcinoma patients after radiotherapy and chemotherapy treatment.

METHODSAccording to treatment modus of nasopharyngeal carcinoma, 64 cases (128 ears) patients were divided to simple radiotherapy group (45 cases, 90 ears) and radiotherapy with chemotherapy group (combination treatment group, 19 cases, 38 ears). Meanwhile, 25 cases (50 ears) people took as control group, who had no obviously ear and nose disease. About two or three months after radiotherapy and chemotherapy treatment completion, three groups were detected by otoscopy, pure tone test, tympanogram and eustachian tube function, respectively. Then, hearing variation of all patients after radiotherapy and chemotherapy treatment were investigated and compared each other. At the same time, the character and level of audiological change were also analyzed.

RESULTSEardrum character of nasopharyngeal carcinoma patients appeared change after radiotherapy. Simple radiotherapy and combination treatment groups were found having hearing impairment and eustachian tube functional disturbance. Moreover, most patients of simple radiotherapy group showed conductive deaf (24%, 22/90), and combination treatment group exhibited mingle (24%, 9/38) or sensorineural deafness (29%, 11/38).

CONCLUSIONSRecent hearing of nasopharyngeal carcinoma patients were damaged by radiotherapy and chemotherapy treatment, radiotherapy treatment induced middle ear or eustachian tube function disturbance, chemotherapy treatment had cochleotoxicity, compared with other treatment, combination treatment was more aggravated hearing impairment.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; Audiometry, Pure-Tone ; Case-Control Studies ; Eustachian Tube ; physiopathology ; Female ; Hearing Loss ; etiology ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; physiopathology ; therapy ; Otitis Media ; etiology ; Radiation Oncology