Dysarthric patients often present velopharyngeal incompetence (VPI), characterized nasalization articulation for hypernasality,which seriously impaired their communication. Research of evaluation of VPI is mainly about cleft palate and postoperative, few about the dysarthria. Some approaches, such as physiologic approach to rehabilitation, have been used to correct hypernasality, and prosthesis,such as palatal lift prosthesis (PLP) and nasal speaking valve (NSV), are also proved effectively. PLP has been widely used for hypernasality oversea, but fewer in China.

Objective To observe the clinical safety of dual axis rotational coronary angiography (DARCA) in the diagnosis of coronary artery disease in Chinese population.Methods From March to December in 2010,74 patients undergoing diagnostic DARCA were enrolled.The improved isocentering technique was adopted in 34 of the patients at the end of the study during DARCA.Blood pressure,heart rate and symptoms were recorded immediately before-and-after contrast injections.Contrast dose,radiation exposure and procedure time for DARCA were recorded.Continuous variable data were analyzed using Student＇s t test,if normality assumption was violated,rank sum test would be used.Categorical variables were analyzed using x2 test.Results ( 1 ) Clinical safety:There was no chest pain documented during or immediately post-injection for all patients.Only 1 patient ( 1 ％ ) had an attack of ventricular tachycardia immediately after the contrast injection and then relieved automatically.Pre and post-injection systolic blood pressure values of left coronary artery were statistically different [ ( 116 ± 20 ) mm Hg vs.( 111 + 18) mm Hg( 1 mm Hg =0.133 kPa),t =3.303,P =0.001 ],and heart rates differed,too [ 73 ( 65- 84)bpm vs.71(64-78) bpm,Z =-4.789,P =0.001 ],but that imposed no clinical significance.(2)Contrast dose,radiation dose and procedure time:The mean contrast utilization,radiation dose and procedure time for DARCA were 28 (25-34) ml,8979 ( 6733-12 363 ) mGycm2 and 200 (164-270) s.Compared with conventional DARCA,improved isocentering technique during DARCA had less radiation exposure and procedure time in left coronary artery angiography and the whole coronary artery angiography [ left coronary artery angiographic radiation exposure:4004 (2932-5772) mGycm2 vs.5808 (4798- 8838) mGycm2,Z =-3.471,P =0.001 ;total radiation exposure:(8116 +2493) mGycm2 vs.( 11 371 ±4122) mGycm2,t =-4.176,P =0.001 ; left coronary artery angiographic procedure time:120 ( 80-180)s vs.150(126-214) s,Z =- 2.836,P =0.005; total procedure time:180 (139-240) s vs.220( 186-308 )s,Z =-3.004,P =0.003 ],but there was no statistically difference in contrast utilization [30(25-35) ml vs.27(25-34)ml,Z=-0.906,P=0.365].Conclusion This study demonstrates clinical safety of DARCA in the diagnosis of coronary artery disease in Chinese population.Compared with conventional isocentering technique of DARCA,improved isocentering technique can significantly reduce radiation exposure and procedure time on the basis of simplified operation,and replace the conventional isocentering technique,but randomized double-blind controlled studies should be conducted.

Objective It aims to investigate the relationships among the categories of Comprehensive Version for Stroke as described in the International Classiifcation of Functioning, Disability and Health (ICF) Core Set, and to provide new supports for Judicial Appraisal of functioning in stroke by ICF functioning mapping.Methods The variables of 59 categories of ICF assessment scale and the samples of 106 persons’ are selected and used in the least absolute shrinkage and selection operator (LASSO) for mining dependencies among those variables. The graphical modeling and analyzing with the software Gephi provides a visual map of the correlations among those classiifcations. Results 59 interconnected categories which organized into the functioning mapping. b340, b735, b175 and b152 are centrally positioned categories because of their high correlation.Conclusion Functioning mapping by graphical modeling can reveal complex relational structures embedded in functioning classiifcations, which provides the support for using ICF to appraisal stroke.

Objective : To evaluate the safety and immunogenicity of a split-virion quadrivalent influenza vaccine. Methods : The healthy people aged three years or over in Wuyang County and Xiping County of Henan Province were divided into the experimental group, control group 1 and control group 2, and were vaccinated with split-virion quadrivalent influenza vaccines, split-virion trivalent influenza vaccines (without B/Victoria) and a split-virion trivalent influenza vaccines (without B/Yamagata) , respectively. The hemagglutination inhibition (HI) antibodies were detected before and after immunization. The incidence rate of adverse events following immunization (AEFI) , HI antibody positive conversion rate, the protection rate of HI antibodies and the growth of geometric mean titer (GMT) were calculated and compared with the standard of Food and Drug Administration (FDA). Results: Totally 2 924 people were recruited, with 975 in the experimental group, 974 in the control group 1 and 975 in control group 2. The incidence rate of AEFI in the experimental group was 11.7%, higher than 7.9% in control group 1 and 8.8% in control group 2 (P < 0.05) during 30 minutes and 8 days after inoculation. The positive conversion rates of HI antibodies of H1N1, H3N2, By and Bv in the experimental group were 78.5%, 53.3%, 78.3% and 62.9%, respectively. The rate differences of the positive conversion rates of HI antibodies of By between the experimental group and control group 2, and of Bv between the experimental group and control group 1 were 42.1% (95%CI: 38.0%-46.2%) and 33.2% (95%CI: 28.9%-37.5%) , with both lower limits of 95%CI more than -0.10. The GMT increase of HI antibodies was more than 2.5 times in the three groups. The protective rates of HI antibodies of H1N1, H3N2, By and Bv in the experimental group were 87.7%, 98.7%, 93.6% and 77.2%, respectively. The protective rates of HI antibodies of By in control group 2 and Bv in control group 1 were 71.1% and 51.0%, both lower than those in the experimental group (P < 0.05). Conclusions After the inoculation of the quadrivalent influenza vaccine, the positive conversion rates (>40%) , protection rates (>70%) and GMT increase (>2.5 times) of HI antibodies of H1N1, H3N2, By and Bv all meet the quality standards of FDA. The safety and immunogenicity of the quadrivalent influenza vaccine are not inferior to those of the trivalent influenza vaccine.

Objective:To analyze the molecular characteristics of Echovirus 11 (Echo11) strains isolated in Xiangyang, Hubei Province from 2016 to 2017 based on the sequences of VP1 gene.Methods:Rectal and throat swab specimens were collected from children with hand, foot and mouth disease (HFMD) in Xiangyang from 2016 to 2017. Echo11 strains were detected by real-time reverse transcriptase PCR (RT-PCR) and isolated after cultured in human rhabdosarcoma (RD) cells. The VP1 regions of Echo11 strains isolated from RD cells and the whole genomes of three representative Echo11 strains were amplified by conventional RT-PCR and the sequences were analyzed. DNAStar7.0 (MegAlign) and MEGA6.0 (Data) were used to analyze the homology and mutation sites in nucleotide and amino acid sequences. Neighbor-joining method was used to construct phylogenetic trees. Recombination analysis was performed with SimPlot software (BootScanning).Results:A total of 11 Echo11 strains were isolated from 3 494 HFMD cases, accounting for 0.31%. They were highly homologous in the VP1 gene. These strains shared 98.4%-100.0% homology in nucleotide sequences and 98.3%-100.0% homology in amino acid sequences. The homology between the 11 Echo11 strains and the prototype strain (Echo11/Gregory, X80059) was 73.9%-74.8% in nucleotide sequences and 87.7%-88.7% in amino acid sequences. All of the Echo11 strains circulating in Xiangyang were classified into lineage D, having a similarity to the strains circulating in some regions of mainland China since 2013. In multiple regions of the genome, the Echo11 strains isolated in Xiangyang were highly similar to the Henan Echo1 strains in 2010 and the Hubei Echo6 strains in 2015, suggesting there was recombination within the genome of Echo11 strains in Xiangyang.Conclusions:The Echo11 strains circulating in Xiangyang from 2016 to 2017 belonged to lineage D and were recombinant strains.

Objective To detect and analyze enteroviruses causing suspected aseptic meningitis in a kindergarten in Jinhua City, Zhejiang Province. Methods Viral RNA was extracted from samples and cDNA was prepared by reverse transcription. PCR was performed to amplify the partial sequences of the 5′-untranslated region ( UTR) and VP1 gene of enteroviruses. Serotypes of the viruses were determined by com-paring the homology between the partial sequences of VP1 gene. Phylogenetic tree of the partial VP1 se-quences was constructed using MEGA6. Results This study included seven patients and twenty-six asymp-tomatic students. Coxsakievirus A10 (CV-A10) was detected in 48. 5％ of the students and echovirus 6 (Echo 6) in 21. 2％. Besides, 12. 1％ of the students might be co-infected by the two viruses. Among the seven patients, six were infected by CV-A10 and the other one might have co-infection. According to the phylogenetic analysis, CV-A10 strains detected in this study were closely related to those isolated in China in recent years, including the strains isolated in Xiamen in 2015 and Yunnan in 2017, while the Echo 6 strains were phylogenetically related to those isolated in Yunnan, Guangzhou and Shandong in 2014. Conclusions CV-A10 and Echo 6 were detected in the cases with suspected aseptic meningitis and had close phylogenetic relationships to the strains appeared in China in recent years.

Objective: To construct the prokaryotic plasmid expressing the recombinant protein Coxsackie virus A6 VP1 or VP2 and prepare the antiserum of rabbit anti-CVA6 VP1 or anti-CVA6 VP2. Methods: CVA6 VP1and VP2 gene fragments were amplified by reverse transcription PCR and inserted into the prokaryotic expression vectors. The recombinant plasmids were expressed in E. coli BL21 (DE3). After induction with Isopropyl β-D-Thiogalactoside (IPTG), the fusion proteins were obtained, and then purified by SDS-PAGE electrophoresis and gel extraction. The polyclonal antibodies were prepared by immunizing rabbits with the fusion proteins and analyzed with Western blot(WB) and indirect immunofluorescence assay(IFA). Results: The prokaryotic expression vector of CVA6 VP1 or VP2 was confirmed by PCR, double enzyme digestion and sequencing. CVA6 VP1 and VP2 fusion proteins with high purity were obtained. WB and IFA were used to identify polyclonal antibodies. Conclusions The CVA6 VP1 and VP2 prokaryotic expression vectors were successfully constructed, and the recombinant CVA6 VP1 and VP2 proteins and their corresponding polyclonal antibodies were obtained.

OBJECTIVETo elucidate the characteristics of genetic variability and its relationship with prevalence, through sequencing and analysis of N gene among street rabies virus isolated from different hosts (homo sapiens, ferret badger, dog) in Zhejiang province.

METHODSSamples were screened and confirmed by direct fluorescence assay and reverse transcript PCR. Sequences were analyzed using bio-information software.

RESULTSEighteen street rabies virus strains were identified, including 2 from homo sapiens, 5 from ferret badger, and 11 from dog. Similarities of N gene and N protein were calculated to be 89.7%-100.0% and 98.4%-100.0% respectively. Mutations occurred in N gene were almost non-sense mutations. In addition,Data from phylogenetic analysis showed that all these strains could be classified into traditional genotype 1.

CONCLUSIONThe prevalence of rabies viruses among different hosts in Zhejiang province had certain regional properties. Rabies viruses isolated from the same kind of host or from the same/adjacent county/counties had the closest relationship. However, the characteristics of rabies virus prevalent in homo sapiens were somewhat complicated. In summary, the transmission of street rabies virus in Zhejiang province was from dogs to ferret badgers and homo sapiens, and the virus could circulate and cross-regional transmit among dogs and ferret badgers.

Animals ; China ; epidemiology ; DNA Mutational Analysis ; Dogs ; virology ; Humans ; Mustelidae ; virology ; RNA, Viral ; genetics ; Rabies ; epidemiology ; Rabies virus ; genetics ; isolation & purification ; Viral Envelope Proteins ; genetics

Objective: To prepare peptide minotope-based recombinant diagnostic antigen of Epstein-Barr virus (EBV) infection and evaluate its antigenicity preliminarily. Methods: With Trx at the N-terminal and His tag at the C-terminal, the peptide minotope of EBV (GP125, F1, A2, A3C2) was expressed in Escherichia coli and purified by affinity and anion exchange chromatography (designated 'H58’); based on antigenicity of H58 identified by Western blotting (WB), we constructed and evaluated a novel early diagnostic ELISA for EBV infection. Results: The soluble H58 protein with high concentration (2.8 mg/ml) and purity (99.01) was obtained; WB analysis found that there was an obvious band (28 ×10³) on the NC membrane, using H58 anti-Trx monoclonal antibody or acute-phase sera of EBV infection as the first antibody. With the novel ELISA, 50 positive sera of EBV infection and 50 negative sera were detected, displaying that the grouping of OD value of positive serum (95%CI: 1.233-1.489) and negative serum (95%CI: 0.113-0.159) was different (P<0.05) with the sensitivity 98.0%, specificity 96.0% and kappa value 0.940. Conclusions By E. coli expression and affinity and ion exchange chromatography purification, the peptide minotope-based recombinant diagnostic antigen of EBV infection was obtained with excellent antigenicity, which could be applied for serological detection of EBV infection.

Objective: To study the relationship between the development of hepatocellular carcinoma(HCC) and HBV gene characteristics among the HCC patients with hepatitis B virus (HBV) infection. Methods: Some acute and chronic hepatitis B patients were collected as control group and HBV associated HCC patients as HCC group. Serum samples of subjects were tested for HBV serological markers. HBV DNA of those samples had been extracted and nested PCR was used to amplify the sequence of HBV DNA. Furthermore, MEGA 6.0 and Bioedit softwares were used to made phylogenetic trees and analyze the gene mutations. Results: The sequences of S region and BCP/Precore region of HBV were amplified from 86 samples in study group and 39 samples in control group. The prevalence of PreS deletion, A1762T and A1762T/G1764A in HCC group were 39.53%, 74.42% and 72.09% respectively, and in control group were 20.51%, 53.85% and 53.85% respectively. The statistical differences of them were significant. The prevalence of A1762T and A1762T/G1764A in ≥ 50 years group were higher than that of < 50 years group. The prevalence of A1762T, G1764A and A1762T/G1764A of subjects who infected genotype C were higher than those infected genotype B. On the contrary, the prevalence of G1896A of subjects who infected genotype C were lower than that of genotype B. It was found that ≥ 50 years, genotype C and G1896A mutation were independently associated with HCC. The risk for suffer from HCC of ≥50 years group, genotype C group and G1896A group were 9.349, 28.875 and 7.648 times compared with < 50 years group genotype B group and without G1896A mutation group, respectively. Conclusions The population of ≥50 years or genotype C had a higher prevalence of A1762T, A1762T/G1764A, ≥50years、genotype C、G1896A were independently associated with HCC, as compared with the subjects of the control group.