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.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.
3.Study on the Mechanism of Aurantii Fructus Immaturus and Its Main Active Ingredients in Promoting Gastrointestinal Motility of Model Rats with Spleen Deficiency
Yuanxiang HU ; Haifang CHEN ; Yupeng SONG ; Shushu TAN ; Xiaoquan LUO ; Wuliang YANG
China Pharmacy 2017;28(13):1747-1750
OBJECTIVE:To study the mechanism of Aurantii fructus immaturus(AFI)and its main active ingredients in pro-moting gastrointestinal motility of model rats with spleen deficiency. METHODS:170 rats were randomly divided into blank group (10 rats) and modeling group (160 rats),rats in modeling group was induced models with spleen deficiency by bitter cold diar-rhea+irregular diet. After modeling, rats were randomly divided into model group, naringin (NA) low-dose, medium-lose, high-dose groups(3.267,6.535,13.070 mg/mL),neohesperidin(NE)low-dose,medium-lose,high-dose groups(3.865,7.730, 15.460 mg/mL),synephrine(SY)low-dose,medium-lose,high-dose groups(0.252,0.504,1.008 mg/mL),compatibility groups with 3 monomer ingredients (NA-NE-SY) low-dose,medium-lose,high-dose and AFI water decoction low-dose,medium-lose, high-dose groups(0.104,0.208,0.416 g/mL,calculated by crude drug),ig,once a day,10 mL/kg,for 7 d. After the last admin-istration,gastrin (GAS) in serum,and acetylcholine (ACh),motilin (MTL),substance P (SP),vasoactive intestinal peptide (VIP)levels in plasma were detected. RESULTS:Compared with blank group,GAS level in serum and ACh,MTL,SP levels in plasma in model group were reduced(P<0.01),VIP level in plasma was increased(P<0.05). Compared with model group,ex-cept for the GAS level in serum showed no obvious change in NA high-dose group and SY doses groups,other medicine groups were obviously increased (P<0.05 or P<0.01);the ACh levels in serum were obviouly increased in NE high-dose group,SY high-dose group and AFI water decoction low-dose group(P<0.01). MTL levels in plasma were obviously increased in NE medi-um-dose,high-dose groups,SY high-dose group,compatibility low-dose,medium-dose groups and AFI water decoction medi-um-dose,high-lose groups (P<0.05);SP levels in plasma were obviously increased in NA low-dose,medium-dose groups and NE doses groups(P<0.05 or P<0.01);VIP levels were reduced in NA low-dose group,SY high-dose group and AFI water decoc-tion low-dose,medium-lose groups(P<0.05). CONCLUSIONS:AFI may promote the gastrointestinal motility of model rats with spleen deficiency by promoting the secretion of GAS,ACh,MTL,and inhibiting the secretion of VIP;there are differences be-tween AFI and the 3 monomer ingredients in regulation of gastrointestinal hormones.
4. 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.
5.Study on the relationship between SNP of CD14 gene and allergic rhinitis in Xinjiang Uygurs and Hans.
Hua ZHANG ; Haifang HU ; Zhilian LIU ; Jie SUN ; Yuping YANG ; Yan WANG ; Guoping TAN ; Yangbing XIANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2011;25(23):1076-1080
OBJECTIVE:
This study aimed to investigate the relationship between CD14 gene rs2569192(C/G), rs3138078 (--1359G/T) and allergic rhinitis (AR) in Xinjiang Uygur and Han populations as well as to determine characteristics of polymorphisms.
METHOD:
A total of 300 AR and 300 healthy controls subjects were included. The frequencies of genotypes and alleles were detected as well as the levels of tIgE in different genotypes were compared.
RESULT:
(1) The distribution of genotypes or alleles of CD14 gene rs2569192 (C/G), rs3138078 (--1359G/T) had no differences between the Xinjiang Uygurs and Hans (P > 0.05). The highest frequency of alleles was C, G. (2) The frequencies of genotypes and alleles were not different between the AR and control group in Uygur and Han (P > 0.05). The frequencies of genotypes and alleles of rs2569192 were different between the Uygur AR and Han AR group (P < 0.05). (3) The distribution of genotype frequencies and allele of rs 2569192 in the Xinjiang Uygur and Han population were quite different from Chinese Beijing Han populations, Japanese, European and African (P < 0.05). (4) The serum total IgE level in AR group was higher than that in healthy control group (P < 0.05).
CONCLUSION
(1) rs2569192 (C/G), rs3138078 (--1359G/T) polymorphisms were not different between the Chinese Xinjiang Uygur and Han population. The major allele were both C and G. rs2569192 of CD14 in Xinjiang populations was different from that in the other populations. (2) No relationship between rs2569192, rs3138078 and AR was found. (3) The serum total IgE level in AR group was higher than that in healthy control group. No relationship between CD14 two SNP and serum total IgE level was found.
Adolescent
;
Adult
;
Aged
;
Aged, 80 and over
;
Alleles
;
Case-Control Studies
;
Child
;
Child, Preschool
;
China
;
epidemiology
;
Ethnic Groups
;
genetics
;
Female
;
Gene Frequency
;
Genotype
;
Humans
;
Hypersensitivity
;
epidemiology
;
genetics
;
Lipopolysaccharide Receptors
;
genetics
;
Male
;
Middle Aged
;
Polymorphism, Single Nucleotide
;
Rhinitis
;
epidemiology
;
genetics
;
Young Adult
6.Gualou Xiebai Banxiatang in Treatment of Cardiovascular Diseases: A Review
Yalong KANG ; Bo NING ; Juanjuan TAN ; Hongfei QI ; Yan SHI ; Fang GUAN ; Haifang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):256-267
Cardiovascular diseases (CVD),a group of common diseases in clinical practice,are witnessing a steady rise in both incidence and mortality rates,posing a challenge to public health. Gualou Xiebai Banxiatang,originating from Synopsis of the Golden Chamber (《金匮要略》),was initially used to treat severe cases of chest impediment. The formula consists of Trichosanthis Fructus,Allii Macrostemonis Bulbus,Pinelliae Rhizoma,and Baijiu. It has a wide range of clinical applications,with therapeutic effects including moving Qi to relieve depression,activating Yang to dissipate mass,and expelling phlegm to alleviate chest congestion. In recent years,clinical research has confirmed that Gualou Xiebai Banxiatang,with or without modification,used alone or in combination with Western medicine,has definite effects in the treatment of CVD such as hyperlipidemia,coronary atherosclerotic heart disease,hypertension,heart failure,and arrhythmia. It can alleviate disease symptoms and reduce the risk of re-hospitalization. Basic research indicates that the mechanisms of Gualou Xiebai Banxiatang include improving endothelial functions,exhibiting anti-inflammatory properties,countering oxidative stress,preventing apoptosis,inhibiting ventricular remodeling,regulating mitochondrial functions,improving hemorheology,and modulating autophagy and neurotransmitters. This article reviews relevant articles in recent years with focuses on the compatibility,clinical application,and mechanism of Gualou Xiebai Banxiatang. This review is expected to provide a theoretical basis for the mechanism research and clinical application of this formula in treating CVD and to offer ideas and reference for in-depth research.
7.Naoqingtong Decoction Ameliorates Kidney Damage in Spontaneously Hypertensive Rats via NLRP3 Inflammasomes
Jiaxin JU ; Caocao CHENG ; Teng GE ; Yalong KANG ; Fang GUAN ; Haifang WANG ; Juanjuan TAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):125-131
ObjectiveTo investigate the effect of Naoqingtong decoction (NQT) on the kidney damage and the inflammatory factors NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC), cysteinyl aspartate-specific proteinase-1 (Caspase-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in spontaneously hypertensive rats (SHRs). MethodsTwenty-four SHRs were randomized into a model group, a low-dose (12.9 g·kg-1·d-1) NQT (NQT-L) group, a high-dose (25.8 g·kg-1·d-1) NQT group (NQT-H), and a captopril (CTP, 20 mg·kg-1·d-1) group, with 6 rats in each group. In addition, 6 homozygous male Wistar-Kyoto rats were used as the control group. The control and model groups were administrated with the same amount of normal saline by gavage for 8 weeks. General behaviors of rats were observed during the intervention period, and the blood pressure was measured periodically. At the end of intervention, the body mass was weighed, and both kidneys were collected and weighed for the calculation of the renal index. Hematoxylin-eosin staining was performed to observe the pathological changes in the kidney tissue. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the expression levels of NLRP3, ASC, Caspase-1, IL-6, and TNF-α in the kidney tissue. ResultsDuring the experiment period, the control group had normal mental status, food intake, and activity, while the model group showed thinning of hair, loss of luster, reduced activity, loss of appetite, fecal adhesion, and irritability, and some of the skin had scratches or blood scabs. The above symptoms were alleviated to different degrees after 8 weeks of NQT administration. An intelligent non-invasive sphygmomanometer was used to measure the tail artery pressure of rats, which showed that the systolic and diastolic blood pressure of rats in the model group was higher than that in the control group (P<0.01). Compared with the model group, drug interventions lowered the systolic and diastolic blood pressure (P<0.05, P<0.01). Compared with the control group, the model group showed severe pathological damage in the kidney tissue, which was alleviated in each drug intervention group. Compared with the control group, the model group showed up-regulated expression levels of NLRP3, ASC, Caspase-1, IL-6, and TNF-α in the kidney tissue (P<0.05, P<0.01). Compared with the model group, the drug intervention groups showed down-regulated expression levels of NLRP3, ASC, Caspase-1, IL-6, and TNF-α in the kidney tissue (P<0.05, P<0.01). ConclusionNQT can lower the blood pressure in SHRs by inhibiting the activation of NLRP3 inflammasomes, suppressing renal inflammation, and ameliorating hypertensive kidney damage.