Objective To evaluate the effect of acupuncture for treating migraine acute attack to offer some evidence‐based basis for clinical application .Methods The Chinese and English literatures on the acupuncture for treating migraine acute attack were retrived from January 1989 to December 2014 ,the literatures were screened according to inclusion and exclusion criteria ,the Meta‐analysis was performed on these chose literatures .Results A total of 5 studies were included and 618 migraineurs were in‐volved ,four literatares were performed the Meta‐analysis ,and 1 literature was performed the description analysis .Meta‐analysis re‐sults showed that there was statistically significant differences between the acupuncture group and the sham acupuncture group in the VAS score reduction value at 2 h[MD=0 .36 ,95% CI:0 .08 ,0 .65 ,P=0 .01] ,4 h[MD=0 .49 ,95% CI:0 .14 ,0 .84 ,P=0 .007] after acupuncture;while when the VAS score was used as the evaluation indicator ,there was no statistically significant differences were found at 2 h[MD= -0 .38 ,95% CI:-0 .83 ,0 .07 ,P=0 .10] ,4 h[MD= -0 .42 ,95% CI:-0 .96 ,0 .12 ,P=0 .12] after acu‐puncture in the VAS score between the acupuncture group and the sham acupuncture group .Conclusion Acupuncture could effec‐tively relieve the intensity of headache in migraine ,the analgesic effect of acupuncture for treating migraine attacks is significantly superior to the sham acupuncture group ,while with the VAS score as the evaluation indicator ,the difference between the acupunc‐ture group and the sham acupuncture group has no statistical significance .

Objective To investigate the possibility of using well-differentiated human airway epithelial cells (HAE) to isolate and identify human influenza A virus from a stale respiratory tract specimen.Methods The stale specimen used in this study was a nasopharyngeal swab specimen collected from a patient with unexplained pneumonia in Qinghai in 2010.It was positive for influenza A virus (H3N2) RNA, but negative for hemagglutination.Equal amount of the specimen was inoculated on HAE and on Madin-Darby canine kidney (MDCK) cells for virus isolation and passage.Cytopathic effects were observed daily after inoculation.Hemagglutination inhibition test was performed at every passage.Electron microscope was used to observe viral morphology.Viral genome was sequenced, followed by molecular evolutionary analysis.Results No progeny virus was isolated in MDCK cells, while a influenza A virus subtype H3N2 strain [A/Qinghai/178/2010(H3N2)] was isolated in HAE with a typical morphology and cytopathic effect of influenza A infection.The hemagglutination inhibition activity was 1∶16.Results of the molecular evolutionary analysis of viral genome showed that the influenza A virus (H3N2) strain was highly homologous to the A/Nanjing/1655/2010(H3N2) strain, which was isolated during the 2010 influenza pandemic in Nanjing.Conclusion HAE can be used for isolation and identification of virus from stale respiratory tract specimens.It is more sensitive than MDCK cells with regard to human influenza virus isolation.

Objective To construct a reverse genetic platform for influenza B virus and to rescue influenza B virus.Methods Eight plasmids carrying the gene segments of B/Florida/4/2006 virus were constructed by using the bidirectional promoter vector pHW2000.293T cells were co-cultured with MadinDarby canine kidney (MDCK) cells and then transfected with the eight plasmids.The supernatants of cell culture and cell debris were collected after transfection and then injected into embryonated chicken eggs and MDCK cells for rescuing the influenza B virus strains.Results This reverse genetic system could be used for the preparation of reassortant influenza B virus strains.The titers of hemagglutination units of the rescued virus achieved 128-256/50μl.Most of the reassortant virus particles were spherical under electron microscope.Conclusion The pHW2000 reverse genetic system could be used for the rescue of influenza B virus.Moreover,it could also be used for the construction of influenza B virus with specific mutations for further in vestigation on the characteristics of influenza B virus and the construction of vaccine strain.

Objective To quantitatively assess the virucidal activities of three commercial disin-fectants against human infected highly pathogenic avian influenza viruses subtype H5. Methods The 50%tissue culture infective dose ( TCID50 ) of avian influenza viruses was calculated. Quantitative suspension test was performed to evaluate the efficacy of three disinfectants. In that test, 105 TCID50 of avian influenza viru-ses were exposed to different disinfectants at different concentrations for different times with or without the in-terference with fetal bovine serum ( FBS) simulating the contaminated condition. The residual infectivity was determined by endpoint titration in Madin-Darby canine kidney ( MDCK) cells. The detail steps were that the mixture of viruses and disinfectants was inoculated at 37℃ with 5% CO2 for 1 hour. Then, it was re-placed by virus dilution medium and further incubated for 18 to 20 hours. ELISA was performed for the cal-culation of TCID50 . The titers of residual viruses were calculated according to Reed and Muench method. The low pathogenic avian influenza virus H9N2 was chosen as the control in this study. Results The re-mained infectivities of three viruses after 1 minute exposure to 1% Virkon solution were below the limit of de-tection (1. 0 lgTCID50/100 μl). Exposing to 0. 5% Virkon solution decreased the viral titers of H5N1 and H9N2 viruses below the detection limit and reduced the titer of H5N6 virus to 1. 75 lgTCID50/100 μl. The virucidal efficacy of 0. 25% Virkon solution against some of the detected viruses was achieved by increasing the exposure time to 5 minutes. The 84 Disinfectant solutions at concentrations of 10%, 5% and 2. 5% low-ered the viral titers of three viruses below the detection limit of 1. 0 lgTCID50/100 μl, but the 1. 25% 84 Disinfectant solution only lowered the viral titers to 1. 25-2. 5 lgTCID50/100 μl. The similar results were ob-served in groups treated with SOLARSEPT solutions. 1% 84 Disinfectant solution didn′t show any virucidal activity against the three viruses after 1 minute of exposure even when the exposure time was extended to 5 minutes. Under the contaminated condition, 1% Virkon solution, 10% and 5% 84 Disinfectant solutions as well as 100% and 50% SOLARSEPT solutions lowered the viral titers below 1. 0 lgTCID50/100μl. Conclu-sion The three commercial disinfectants (1% Virkon solution, 10% 84 Disinfectant solution and SOLAR-SEPT solution) were efficient virucides for highly pathogenic avian influenza viruses subtype H5 even under the contaminated condition. Increasing the exposure time had no significant effects on the efficacy of three disinfectants after the virucidal activities were neutralized by enough viruses. No significant differences in vi-rucidal activities of three disinfectants against HPAI H5 viruses and LPAI H9 virus were observed.

Preparation of maternal strain A/PR/8/34 HA antiserum for influenza virus classical reassortment. A/PR/8/34 virus was digested by bromelain after inactivation and purification. 5%-20% sucrose continuous density gradient centrifugation method was used to purify HA protein. SIRD method was used to select the target protein. SDS-PAGE method was used to identified HA protein. High Immunogenic A/PR/8/34 HA protein was successfully prepared and HI titer reached 10240. High purity HA antiserum was identified by SIRD method. The key reagent in the classical reassortment of influenza virus was prepared, and the complete set of technical methods were explored, which laid the foundation for the independent research and development of seasonal influenza vaccine strains of China.
Animals ; Antibodies, Viral ; immunology ; Electrophoresis, Polyacrylamide Gel ; Female ; Hemagglutination Inhibition Tests ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; immunology ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Rabbits ; Reassortant Viruses ; genetics ; immunology

Abstract: To investigate the distribution of avian influenza virus in environmental samples from live poultry markets (LPM) in China, samples were collected and tested by nucleic acid during 2009-2013 season. Each sample was tested by real-time RT PCR using flu A specific primers. If any real-time PCR was positive, the sample was inoculated into specific-pathogen-free (SPF) embryonated chicken eggs for viral isolation. The results indicated that the positive rate of nucleic acid in enviromental samples exhibited seasonality. The positive rate of nucleic acid was significantly higher in Winter and Spring. The positive rate of nucleic acid in LPM located in the south of China was higher than in northern China. Samples of Sewage for cleaning poultry and chopping board showed that higher positive rate of nucleic acid than other samples. The Subtype identification showed that H5 and H9 were main subtypes in the enviromental samples. Viral isolation indicated H5 subtypes was more than H9 subtypes between 2009 and 2013 while H9 subtypes increased in 2013. Our findings suggested the significance of public health based on LPM surveillance and provided the basis of prevention and early warning for avian flu infection human.
Animals ; China ; Feces ; virology ; Fresh Water ; virology ; Influenza A virus ; classification ; genetics ; isolation & purification ; Influenza in Birds ; virology ; Poultry ; Poultry Diseases ; virology ; Public Health ; Seasons ; Sewage ; virology

Objective:To investigate the effects of pre-existing antibody on seroconversion rate after influenza vaccination.Methods:This study recruited 1 900 healthy volunteers to receive influenza split vaccines in Xinjiang Uygur Autonomous region and Yunnan Province from September 2009 to October 2018. Hemagglutinin agglutination inhibition assay was used to detect the titers of specific antibodies in blood samples collected before vaccination and 28 d after vaccination and the effects of pre-existing antibody on the seroconversion to different influenza vaccine components were analyzed.Results:Trend analysis showed that with the increasing titer of pre-existing antibody, the seroconversion rates to A/H1N1, A/H3N2, B/Victoria and B/Yamagata vaccine components were gradually decreased (χ 2=121.76, P<0.001; χ 2=67.58, P<0.001; χ 2=45.25, P<0.001; χ 2=54.55, P<0.001). After adjusting for factors such as region, gender and age, multivariate logistic regression showed that pre-existing antibody titer equal to or higher than 40 was an independent factor that affected the seroconversion to A/H1N1, A/H3N2 and B/Victoria vaccine components, and the adjusted OR (95%CI) values were 2.50(2.00-3.13)、1.64(1.35-2.00) and 2.50(1.79-3.45), respectively. Conclusions:The seroconversion rate to each vaccine component was negatively correlated with the pre-existing antibody titer. The factor that pre-existing antibody titer equal to or higher than 40 was detrimental to the seroconversion to A/H1N1, A/H3N2 and B/Victoria vaccine components, but had no significant influence on B/Yamagata seroconversion.

Data based on the antiviral-resistant phenotyping characteristics of 884 influenza B viruses circulating in mainland China from October 2013 to March 2014 were analyzed to assess the susceptibility of influenza B viruses to neuraminidase inhibitors. All 884 viruses were sensitive to oseltamivir; two viruses (0.23%) had reduced sensitivity to zanamivir and all other viruses were sensitive to zanamivir. Among the 38 viruses with a B/Victoria lineage, B/Shandong-Kuiwen/1195/2014 exhibited a half-maximal inhibitory concentration (IC50) for zanamivir that was elevated by 5. 12-fold (1.78 nM) compared with neuraminidase inhibitors sensitive to the reference virus (0.34 nM), suggesting that it exhibited reduced inhibition by zanamivir. D35G, N59D and S402T (39, 64 and 399 with N2 number) amino-acid substitutions in the NA gene were detected with no previously reported antiviral-resistant substitutions. Among viruses with the 846 B/Yamagata lineage, B/Hunan-Lingling/350/2013 exhibited a 7.99-fold elevated IC50 for zanamivir (2.72 nM) compared with neuraminidase inhibitors sensitive to the reference virus (0.34 nM), suggesting that it exhibited reduced inhibition by zanamivir. D197N (N2 number), a previously reported antiviral resistant-related amino-acid substitution in the NA gene, was detected in B/Hunan-Lingling/350/2013. These data suggest that recently circulating influenza B viruses in mainland China have retained susceptibility to neuraminidase inhibitors.
Amino Acid Substitution ; Antiviral Agents ; pharmacology ; China ; epidemiology ; Drug Resistance, Viral ; Enzyme Inhibitors ; pharmacology ; Humans ; Influenza B virus ; drug effects ; enzymology ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Microbial Sensitivity Tests ; Neuraminidase ; antagonists & inhibitors ; genetics ; metabolism ; Viral Proteins ; antagonists & inhibitors ; genetics ; metabolism

Objective To analyze the phenotypic characteristics of antiviral-resistant influenza B viruses circulating in mainland China and to analyze the susceptibility of influenza B viruses to neuraminidase inhibitors ( NAIs) . Methods Antiviral-resistant phenotyping test was performed to analyze the NAI suscep-tibility of 1 386 influenza B viruses isolated in mainland China from April 2014 to March 2015, including the test of susceptibility to oseltamivir and zanamivir. Results All of the 94 B-Victoria lineage viruses were sensitive to oseltamivir and zanamivir. Of all 1 292 B-Yamagata lineage viruses tested, 1 virus showed re-duced sensitivity to oseltamivir with NA gene containing I221T amino acid mutation, 10 viruses showed re-duced sensitivity to zanamivir with 4 having D197N amino acid mutation in NA gene, 3 viruses showed re-duced sensitivity to both oseltamivir and zanamivir with NA gene possessing D197N amino acid mutation and 1 virus carrying the A245T amino acid mutation in NA gene showed reduced sensitivity to oseltamivir and highly reduced sensitivity to zanamivir. Conclusion The majority of influenza B viruses circulating in main-land China during 2014 to 2015 were sensitive to NAIs, which indicated that NAIs could be used continually for clinical treatment of patients with influenza. Sustained monitoring of antiviral susceptibility of influenza B viruses should be emphasized for timely detection of antiviral resistant viruses and more attention should be paid to the D197N mutations in NA gene of influenza B viruses.

OBJECTIVE: To explore the genetic basis for a pedigree affected with KBG syndrome. METHODS: Clinical data of three patients from the pedigree (the proband, his mother and sister) was collected. Genomic DNA was extracted from peripheral blood samples and subjected to whole exome sequencing (WES). Suspected variant was verified by Sanger sequencing. RESULTS: The proband was found to harbor a heterozygous c.4398_4401del (p.Glu1467AsnfsTer63) frameshift variant of the ANKRD11 gene by WES. Sanger sequencing confirmed that the same variant was also present in his mother and sister, but not in his father. CONCLUSION The c.4398_4401de (p.Glu1467AsnfsTer63) variation of the ANKRD11 gene probably underlies the KBG syndrome in this pedigree.