1.Analysis on gene E sequence of Dengue type Ⅰ virus strains in Zhaoqing during 2014
Haifang TAN ; Dawei GUAN ; Hanqing TAN
International Journal of Laboratory Medicine 2017;38(3):370-372
Objective To understand the gene E sequences of prevalent strains of Dengue fever type Ⅰ virus in Zhaoqing City during 2014.Methods The medical record data and acute stage serum of the patients with Dengue fever in Zhaoqing City during 2014 were collected.Dengue virus was cultured and isolated by using the C6/36 cell culture.Gene E of positive strains was amplified with RT-PCR.The phylogenetic tree was drawn and the bioinformatics analysis was conducted.Results Of 36 samples,20 samples were positive in viral isolation and culture.The gene E sequences of 20 strains of type Ⅰ Dengue virus prevalence in Zhaoqing dur-ing 2014 were obtained;the homology of these sequences was close to that of the 2 prevalent strains found in Zhongshan City,but was distant from that found in Guangzhou City.Conclusion The epidemic situation of Dengue fever in Zhaoqing City is closely re-lated to the prevalence situation of Zhongshan and is characterized by imported prevalence.
2.An analysis of adverse drug reactions of thalidomide in treatment of immune-related bowel diseases
Hanqing LUO ; Bei TAN ; Hong Lü ; Jiaming QIAN
Chinese Journal of Internal Medicine 2013;52(9):726-729
Objective To investigate the safety of thalidomide in the treatment of immune-related bowel diseases for providing clinical reference.Methods Thirty-five patients with immune-related bowel diseases (31 Crohn's disease,2 ulcerative colitis and 2 Behcet's disease) treated with thalidomide were enrolled in this study.The incidence,type,severity,duration of thalidomide related adverse drug reaction (ADR) and the dose-effect relationship of neurotoxicity were analyzed.Results All the patients were treated with a mean dose of thalidomide (109.29 ± 30.37) mg/d for (18.8 ± 12.4) months,and 33 occurred ADR.The three most frequent ADR were numbness [51.4% (18/35)],somnolence [48.6% (17/35)] and dermatitis [37.1% (13/35)].The median time to development of these three ADR were 6.50,0.25,and 1.00 months,respectively.Severe ADR leading to withdrawal accounted for 20.0% (7/35),including reasons of peripheral neuritis (3/7),dermatitis (2/7) and myelosuppression (2/7).The incidence of peripheral neuritis was not significantly related to the maximal and initial dose of thalidomide (P > 0.05).Conclusions Although the incidence of ADR was relatively high during the treatment of thalidomide,most of them were mild and well tolerated.Thalidomide can be safely used in patients with immune-related bowel diseases under close monitoring.
3.Genomic characterization of avian influenza A(H7N9) virus in Zhaoqing, China, 2014-2016
Hanqing TAN ; Jieping CHENG ; Haifang TAN ; Yingmei ZHU ; Feng LIN ; Xiaoqing LIANG ; Jie CHEN ; Wei MAI
Chinese Journal of Zoonoses 2017;33(3):202-207,240
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.
4.Efficacy and safety of thalidomide in the treatment of 29 refractory Crohn's disease patients
Hanqing LUO ; Yue LI ; Hong LYU ; Zhanghan DAI ; Bei TAN ; Jiaming QIAN
Chinese Journal of Digestion 2016;36(3):172-176
Objective To evaluate the efficacy and safety of thalidomide (100 to 200mg per day) in the treatment of adult refractory Crohn's disease (CD).Methods From July 2008 to February 2013,29 refractory CD patients were enrolled in thalidomide (100 to 200 mg per day)cohort study.The clinical activity was evaluated by simplified CD activity index.Patients in clinical remission underwent colon endoscope examination,and mucosal healing was assessed by simple endoscopic score for Crohn's disease (SES-CD).Adverse reactions (ADR) were also observed.Results Among the 29 CD patients,23 males and six females,the baseline of 19 patients (65.5%) were in clinical active period and 10 in clinical remission period.Among patients with baseline in clinical active period,three patients did not reach the target dose because of ADR,the left 16 patients were treated with thalidomide for one year and 14 patients achieved clinical remission.The median time of inducing clinical remission was one month.A totle of 24 patients with clinical remission induced by thalidomide and with baseline in clinical remission period were assessed in efficacy evaluation of mucosal healing.Thalidomide was withdrawn in three patients in six months because of ADR and colonoscopy evaluation did not complete,while the other 21 patients received colonoscopy evaluation among whom 33.3% (7/21) achieved mucosal healing after two years of thalidomide treatment.Numbness of the hands,feet or mouth,somnolence and dermatitis were the top three ADR of thalidomide treatment.A total of nine patients withdrew the medication because of ADR (four (13.8%) with numbness of the hands,feet or mouth,four (13.8%) with dermatitis and one (3.4%) with leukopenia).Conclusions Thalidomide 100 to 200 mg per day can induce clinical remission and mucosal healing in refractory CD.However,it has some adverse reactions and close monitoring and follow up are required during treatment.
5.Study on the value of PCR for rapid screening Salmonella/Shigella in preventive health physical examination
International Journal of Laboratory Medicine 2017;38(23):3268-3269,3273
Objective To investigate the clinical value of PCR method for rapid screening Salmonella/Shigella in health physical examination .Methods A total of 3256 cases who were taken health physical examination from July 2011 to June 2015 ,were select-ed as the research object .Real-time fluorescence quantitative PCR method and the culture method were used for detecting Salmonel-la/Shigella ,and two methods were compared on their strengths and weaknesses for screening Salmonella/Shigella .Results A total of 46 cases with Salmonella were detected by PCR ,and the ratio was 1 .41% ;41 cases with Salmonella were detected by traditional culture method ,and the ratio was 1 .26% .39 cases with Shigella were detected by PCR and ,the ratio was 1 .20% ;32 cases were de-tected by traditional culture method ,and the ratio was 0 .98% .The difference was statistically significant (P<0 .05) .Sensitivity and specificity of PCR to detect Salmonella were 100 .00% and 99 .84% ,respectively ,and sensitivity and specificity of PCR to detect Shigella were 100 .00% and 99 .78% ,respectively .The PCR method is better than the traditional culture method concerning man-power ,processing time ,and satisfaction of test-takers ,and the cost of PCR method is significantly higher than that of traditional culture ,with a significant difference (P<0 .05) .PCR method contains high technology and has strong objectivity .Conclusion Real time fluorescent quantitative PCR method ,with high efficiency ,can be employed well for screening intestinal tract pathogenic bacte-ria ,improve the detection rate and accuracy for detecting Shigella and Salmonella ,and save time and energy as well .
6. Development and application of dual real time RT-PCR for avian influenza H5N6 virus
Hanqing TAN ; Jieping CHENG ; Yingmei ZHU ; Haifang TAN ; Qiang HUANG ; Lebin SU ; Feng LIN ; Tingguo DENG ; Bijian LI
Chinese Journal of Experimental and Clinical Virology 2017;31(1):62-65
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.
7.Association of aggressive behaviors of schizophrenia with short tandem repeats loci.
Chun YANG ; Huajie BA ; Xingqi TAN ; Hanqing ZHAO ; Shuyou ZHANG ; Haiying YU
Chinese Journal of Medical Genetics 2017;34(6):901-904
OBJECTIVETo assess the association of short tandem repeats (STRs) loci with aggressive behaviors of schizophrenia.
METHODSBlood samples from 123 schizophrenic patients with aggressive behaviors and 489 schizophrenic patients without aggressive behaviors were collected. DNA from all samples was amplified with a PowerPlex 21 system and separated by electrophoresis to determine the genotypes and allelic frequencies of 20 STR loci including D3S1368, D1S1656, D6S1043, D13S317, Penta E, D16S639, D18S51, D2S1338, CSF1PO, Penta D, TH01, vWA, D21S11, D7S820, D5S818, TPOX, D8S1179, D12S391, D19S433, and FGA.
RESULTSAll of the 20 STR loci have reached Hardy-Weinberg equilibrium in both groups. A significant difference was found in allelic and genotypic frequencies of loci Penta D between the two groups (alleles: P=0.042; genotypes: P=0.014) but not for the remaining 19 loci (P> 0.05). Univariate analysis also showed a significant difference for allele 10 and genotypes 10-12 of Penta D between the two groups (P=0.0027, P=0.0001), with the OR being 1.81 (95%CI: 1.22-2.67) and 4.33 (95%CI: 1.95-9.59), respectively.
CONCLUSIONPenta D may be associated with aggressive behaviors of schizophrenia. Allele 10 and genotypes 10-12 of Penta D may confer a risk for the disease.
Adolescent ; Adult ; Aged ; Aggression ; Gene Frequency ; Genotype ; Humans ; Microsatellite Repeats ; Middle Aged ; Schizophrenia ; genetics ; Young Adult
8.Expression of Twist1, SIRT1, FGF2 and TGF-β3 genes and its regulatory effect on the proliferation of placenta, umbilical cord and dental pulp mesenchymal stem cells.
Yao TAN ; Yin DENG ; Keyou PENG ; Zhengzhou SUN ; Jianqiu HUANG ; Xuntong GU ; Fusheng ZHANG ; Hanqing PENG ; Xuechao ZHANG ; Rong ZHANG
Chinese Journal of Medical Genetics 2021;38(2):117-122
OBJECTIVE:
To compare the mRNA level of cell proliferation-related genes Twist1, SIRT1, FGF2 and TGF-β3 in placenta mesenchymal stem cells (PA-MSCs), umbilical cord mensenchymals (UC-MSCs) and dental pulp mesenchymal stem cells (DP-MSCs).
METHODS:
The morphology of various passages of PA-MSCs, UC-MSCs and DP-MSCs were observed by microscopy. Proliferation and promoting ability of the three cell lines were detected with the MTT method. Real-time PCR (RT-PCR) was used to determine the mRNA levels of Twist1, SIRT1, FGF2, TGF-β3.
RESULTS:
The morphology of UC-MSCs and DP-MSCs was different from that of PA-MSCs. Proliferation ability and promoting ability of the PA-MSCs was superior to that of UC-MSCs and DP-MSCs. In PA-MSCs, expression level of Twist1 and TGF-β3 was the highest and FGF2 was the lowest. SIRT1 was highly expressed in UC-MSCs. With the cell subcultured, different expression levels of Twist1, SIRT1, FGF2, TGF-β3 was observed in PA-MSCs, UC-MSCs and DP-MSCs.
CONCLUSION
Up-regulated expression of the Twist1, SIRT1 and TGF-β3 genes can promote proliferation of PA-MSCs, UC-MSCs and DP-MSCs, whilst TGF-β3 may inhibit these. The regulatory effect of Twist1, SIRT1, FGF2 and TGF-β3 genes on PA-MSCs, UC-MSCs and DP-MSCs are different.
Cell Differentiation
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Cell Proliferation/genetics*
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Cells, Cultured
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Dental Pulp/cytology*
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Female
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Fibroblast Growth Factor 2/genetics*
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Humans
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Mesenchymal Stem Cells/cytology*
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Nuclear Proteins/genetics*
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Placenta/cytology*
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Pregnancy
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Sirtuin 1/genetics*
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Transforming Growth Factor beta3/genetics*
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Twist-Related Protein 1/genetics*
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Umbilical Cord/cytology*