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Objective@#To establish a TaqMan-MGB probe-based real-time fluorescence RT-PCR assay for avian influenza H5N6 virus used in rapid diagnosis for suspected cases and surveillance for outer environment of live poultry markets.@*Methods@#Based on the conservative sequences of avian influenza H5N6 virus for HA and NA gene published on GenBank, specific primers and TaqMan-MGB probes were designed to develop and optimize for the dual real-time RT-PCR assay. Specificity, sensitivity, repeatability and comparison tests were carried out.@*Results@#This dual real-time RT-PCR detection can be completed within 80 minutes. There was no cross-reaction with other subtypes of influenza virus and common respiratory pathogens. The minimum detection limit could be up to 10 copies/reaction. The correlation coefficient of standard curve for the gene of H5 and N6 were 0.999 and 0.993, and the coefficients of variation for cycle threshold were range from 0.151%-0.549%and 0.213%-0.575%, respectively. The positive and negative coincidence rates of the validation test were 100%.@*Conclusions@#This TaqMan-MGB probe-based dual real-time RT-PCR for avian influenza H5N6 virus was rapid, specific and sensitive. It will have a good use in early emergency detection of suspected cases and continuous monitoring of external environment in live poultry trade market.
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We analyzed genetic evolution characteristics of avian influenza A (H7N9) virus isolated in Zhaoqing,China,2014-2016.Nucleic acid were extracted and sequenced from 17 samples of H7N9 positive cases in Zhaoqing.Genetic characteristics of homology and important amino acid sites were analyzed by using BioEdit5.0 and MEGA6.0.The evolutionary trees were constructed by Neighbor-Joining and the referenced sequences were downloaded from GenBank,Eight nucleic acid fragments from 7 strains of H7N9 viruses were successfully generated.The highest homology was found in HA gene with A/chicken/Dongguan/695/2014(H7N9),and NA gene with A/chicken/Dongguan/1075/2014(H7N9).The internal genes were high homology with avian H7N9 and H9N2 virus from Dongguan and Shenzhen in Guangdong,China.The HA and NA genes were directly evolved in the Pearl River Delta evolution branch with the H7N9 sequences from the cities of Dongguan,Guangzhou and Shenzhen,while the sequences from the provinces of Anhui,Zhejiang,and Jiangsu were in the Yangtze River Delta evolution branch.There were 2 alkaline amino acids in cleavage site of HA,2 mutations (G186V and Q226L) in the crucial sites related with the receptor of HA protein,1 mutation (E627K) in PB2 protein,and 1 drug resistance mutation (S31N) in M2 protein.And no evidence of neuraminidase resistance in NA protein was found.In conclusion,the H7N9 virus for human infection in Zhaoqing may originate from avian H7N9 and H9N2 viruses,which circulated in the Pearl River Delta region of Guangdong from 2013 to 2014.The mutations of G186V,Q226L and E627 K might be related with high susceptibility to human beings.
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BACKGROUND:Few reports are found about the effect of ovariectomized rats' bone histomorphometry parameters using non-destructive dynamic loading system.OBJECTIVE:To observe the effect of different loads situations on the bone histomorphometry parameters in ovariectomized rats.DESIGN,TIME AND SETTING:A controlled observation on the bone histomorphometry was performed in the Biomedical Engineering Laboratory of Jilin University from April 2007 to August 2007.MATERIALS:A total of 35 9-month-old Sprague-Dawley female rats were randomly divided into five groups,including sham operation group,ovariectomized (OVX) control group,OVX loading 1 N group,OVX loading 2 N group,OVX loading 4 N group.There were 7 rats in each group.METHODS:Rats in OVX control group and castration load group were processed into bilateral OVX on the back. The sham operation group only underwent the excision of fat tissues on back,and then sutured. After castration for 1 week,rats were loaded with non-destructive dynamic loading system in the two sides of the tibia,15 minutes a day. The mechanical loads would continue for 4 weeks and the loads were 1N,2N and 4N.MAIN OUTCOME MEASURES:Changes of proximal tibia bone histomorphometry parameters.RESULTS:The area,number and thickness of trabecular bone in OVX loading group were all higher than OVX control group (P＜0.05,P＜0.01).The trabecular bone area and thickness in OVX 4 N and OVX 2 N groups were significantly higher than OVX control group (P ＜ 0.001).There was a downward trend of trabecular separation in OVX 4 N group compared with OVX control group (P ＜ 0.05). With the increasing loads,there was an increasing trend of the area,number and thickness of trabecular bone,which were close to sham-operated group. The trabecular separation was declined. Single fluorescent labeled surface and double fluorescent labeled surfaces in sham operated group were all lower than that in OVX control group. With the increase in loads,the single fluorescent labeled surface,double fluorescent labeled surface,interlabeled width and mineral apposition rate had been shown to increase. The OVX 2 N and OVX 4 N groups exhibited a remarkably higher level of mineral apposition rate than OVX control group (P ＜ 0.05).CONCLUSION:With the increase in load at the range of 1-4 N,all parameters of bone histomorphometry improve in the OVX rats,the bone microstrcture is greatly ameliorated,bone mass loss is reduced and the process of osteoporosis is delayed.
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@# Objective To investigate the therapeutic effect of percutaneous laser disc decompression on lumbar discogenic pain.Methods56 patients with lumbar discogenic pain were treated with percutaneous laser disc decompression with Nd:YAG laser(wavelength 1064 nm).They were followed up for more than 3 months with Macnab criteria.ResultsAll the patients were followed up.At the end point of postoperative 3 months,32 patients had an excellent outcome,20 were good,3 patients were fair,1 patient were poor.No complication(infection and nerve injury)had been observed.ConclusionPercutaneous laser disc decompression is a safe,little invasive and effective treatment modality for lumbar discogenic pain.
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Objective:To study the effects of pulse electro-magnetic fields (PEMFs) on the healing of tibial condylar osteochondral in rabbits.Methods:Sixteen rabbits were divided into two groups,and osteochondral fractures of left tibia condyle were created,internal fixation was performed with steel wire.Group 1 was constantly treated with PEMFs six hours per day;group 2 was not treated.The results were studied by gross morphology,light microscopy and elect ro-microscopy.Results:75% and 13% fractures were repaired by hyalin-like cartilage in group 1 and in group 2 respectively.Conclusion:PEMFs are essential to facilitation of the cartilage repair.