1.Bioinformatic analysis of regulation of microRNA on target genes in pediatric asthma
Xiaoyan DONG ; Quan LU ; Huiyan ZHANG ; Jianlei GU ; Nan ZHONG
Journal of Clinical Pediatrics 2016;(2):81-87
Objective To understand the underlying mechanism of mites-induced pediatric asthma by bioinformatic analysis on speciifc microRNA (miRNA) array and target gene screening. Methods This is a case control study of 62 pairs of dust mites-induced asthma children with age and gender matched healthy controls. Twelve pairs were randomly selected for miRNA array. The abnormal expression of miRNAs was compared between asthma and control children. The results were validated by RT-qPCR and bioinformatic analysis in remaining pairs of children. Results Six miRNAs (miRNA-151a-5p, 625-5p, 126-3p, 513a-5p, 27b-3p, 22-3p) were signiifcantly down-regulated more than two folds in dust mites-induced asthma children than those in controls. The enriched bioinformatics analysis showed that these miRNAs and their target genes CBL, PPARGC1B, ESR1, ONECUT2, EGFR, SYK, and STAT1 were related to inlfammatory cytokine signaling pathway. Conclusion It is suggested that miR-22-3p, 513a-5p, 625-5p, 27b-3p, and miRNA-target genes form a network through co-regulation to target genes to participate dust mites-induced asthma in children.
2.The comparison of establishment of the acute myocardial infarction model between the beagle dogs and the mini-pigs by interventional technique
Xiaolong GU ; Jun HUANG ; Zhenghua DONG ; Huiyan LEI ; Zhihua GONG ; Yanfei WEN ; Dingcheng XIANG
Chongqing Medicine 2014;(9):1090-1092
Objective To explore the success rate and the risk of establishment of the acute myocardial infarction model between the beagle dogs and the mini-pigs by interventional technique ,further to provid theoretical basis for choose a more suitable animal model .Methods 6 dogs and 6 mini-pigs were anaesthetized ,then underwent the coronary arteriography via femoral artery .After is-chemic preconditioning the coronary balloon was inflated to occlude the middle left anterior descending coronary for 180 minutes . The electrocardiogram was examined throughout the operation and the pathological sections were examined until the animals were executed one week later .Results All beagle dogs survived ,while 1 case of mini-pigs dead(1/6) .There was 1 cases(1/6) of beagle dogs had acute myocardial infarction ,while 5(5/6)cases in mini-pigs .All mini-pigs had malignant arrhythmia(6/6) but never seen in beagle dogs .The time needed for building a model was similar between the two groups ,the difference had no statistical signifi-cance(P>0 .05) .Conclusion The risk of establish myocardial infarction model in mini-pigs is higher than beagle dogs ,but the suc-cess rate is still high ,it might be the better choice .
3.Expression and immunization assessment of HSV-2 glycoprotein D in baculovirus expression vector system
Wei LIU ; Xiaohua LUO ; Wenjing CHEN ; Yuan DONG ; Jie ZHU ; Jian GUO ; Yong JIANG ; Lei ZHANG ; Guixian MENG ; Huiyan WANG
Journal of Jilin University(Medicine Edition) 2017;43(4):720-724
Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.
4.Analysis of Thyroid Carcinoma Animal Model Based on Clinical Characteristics of Chinese and Western Medicine
Yike AN ; Mengfan PENG ; Huiyan YANG ; Hongyang DONG ; Mingsan MIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(4):951-956
Objective To summarize the modeling methods and standards of thyroid cancer,to provide reference for the establishment of thyroid cancer animal model close to the clinical syndrome of Chinese and Western medicine,and to promote the progress of clinical diagnosis and treatment.Methods The modeling methods and characteristics of animal models of thyroid cancer in CNKI,Wanfang,Web of Science and PubMed databases were analyzed and summarized.The advantages and disadvantages were evaluated,and the coincidence degree with the clinical characteristics of traditional Chinese and Western medicine was analyzed.Results There are many modeling methods for thyroid cancer animal models,which are mainly divided into four types:spontaneous,induced,genetic engineering and transplantation models.Genetic engineering mouse models include transgenic models,gene knockout models,gene replacement models and the latest reported restricted mouse models.Among them,the genetic engineering mouse model and the transplanted mouse model have a high clinical coincidence,and the spontaneous and induced tumor model have a low clinical coincidence.Conclusion It is one of the important directions for future research on thyroid cancer to establish an animal model consistent with"TCM syndrome"and"Western medicine diagnostic criteria".
5.mRNA expression of DNA methyltransferase 2 and 3a in the epidermis of patients with psoriasis vulgaris
Xuhua MAO ; Chao LUAN ; Yu HU ; Huiyan DONG ; Mingjun JIANG ; Min CHEN
Chinese Journal of Dermatology 2018;51(8):607-608
Objective To investigate the expression of DNA methyltransferase 2 (DNMT2) and 3a (DNMT3a) in the epidermis of patients with psoriasis vulgaris.Methods Between March 2009 and December 2010,46 patients with psoriasis vulgaris were enrolled from the Department of Dermatology,Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College,and the Department of Dermatology of Yixing People's Hospital,and 28 healthy controls were enrolled from the Department of Dermatologic Surgery,Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College.Real-time quantitative PCR was performed to determine the mRNA expression of DNMT2 and DNMT3a in the epidermis of the lesional and nonlesional skin of patients with psoriasis vulgaris,as well as in the epidermis of normal skin of the healthy controls.Results The mRNA expression of DNMT2 (expressed as 2-△△Ct) in the lesional skin,non-lesional skin of the patients and normal skin of the healthy controls was 0.62 ± 0.02,0.36-± 0.05 and 0.15 ± 0.11,respectively.The mRNA expression of DNMT2 was significantly higher in the lesional skin than in the non-lesional skin of the patients (t =6.23,P < 0.01),and higher in the non-lesional skin of the patients than in the normal skin of the healthy controls (t =7.33,P < 0.01).Additionally,the mRNA expression of DNMT3a was significantly higher in the lesional skin (0.85 ± 0.03) than in the non-lesional skin (0.43 ± 0.04) of the patients (t =5.66,P < 0.01),and higher in the non-lesiona] skin of the patients than in the normal skin of healthy controls (0.18 ± 0.09,t =8.62,P < 0.01).Conclusion Both DNMT2 and DNMT3a mRNA were abnormally expressed in the epidermis of patients with psoriasis vulgaris.
6.Inhibitory effect of proliferation and promotion effect of apoptosis induction of T-2 toxin on human hepatocellular carcinoma HepG2 cells
Wenhe ZHU ; Yu LIU ; Jiaqi ZHANG ; Yan LI ; Wei LIU ; Yuan DONG ; Lei LIU ; Huiyan WANG
Journal of Jilin University(Medicine Edition) 2017;43(6):1087-1091,前插2
Objective: To investigate the inhibitory effect of T-2 toxin on proliferation of the human hepatocellular carcinoma HepG2 cells and its promotion effect of apoptosis.Methods:The HepG2 cells in the logarithmic phase were selected and divided into control group (without T-2 toxin)and experimental groups (given 0.25,2.50,25.00,250.00 and 2500.00 μg·L-1 T-2 toxin).After 24 h treatment,the morphology of cells was observed under inverted microscope;the inhibitory rate of proliferation of cells was determined by MTT assay;the cell cycle and apoptotic rate of cells were analyzed by flow cytometry;Hochest 33258 staining was used to observe the apoptotic morphology of cells;the activity of caspase-3 in HepG2 cells was detected.Results:After treated for 24 h,the inverted microscope observation results showed that the number of the cells in experimental groups was decreased significantly and the cells shrank and deformed.The MTT results showed that compared with control group,the inhibitory rates of proliferation of the cells in experimental groups were increased (P <0.01).The flow cytometry results showed that compared with control group, the percentage of the cells in SubG1 phase in experimental group was significantly increased,and the apoptotic rates of the cells in experimental groups were significantly increased.The Hoechest 33258 staining results showed that the chromatin condensation was observed in the cells in experimental groups,and the nuclei were dense and stained.Compared with control group,the activities of intracellular caspase-3 of the cells in experimental groups were significantly increased (P < 0.01 ). Conclusion:T-2 can inhibit the proliferation of human hepatocellular carcinoma HepG2 cells,and induce the apoptosis.
7.Effects of group practice health management on negative emotions of polycystic ovary syndrome patients
Huiyan JU ; Pan DOU ; Jing SHANG ; Fei CHEN ; Li DONG ; Na ZHANG ; Xueying LI ; Yang XU
Chinese Journal of Modern Nursing 2016;22(34):4959-4961,4962
Objective To investigate the effect of polycystic ovary syndrome( PCOS) group practice on the patients′negative emotions, and aim to help PCOS patients improve emotions, establish healthy lifestyle and improve the reproductive function, as well as to lay basis for the health care management model of PCOS group practice. Methods In this study, 56 patients whose percentage of body fat ( BF%) was more than 30% were included in this PCOS group practice. We spent 8 to 12 weeks on the health management, and compared the negative emotions before and after the health management. Results Before the health management, the incidence rate of anxiety was 78.6%, the score of self-rating anxiety scale (SAS) was (45.33±7.94), the incidence rate of depression was 51.8%, and the score of self-rating depression (SDS) was (0.49±0.11). After the health management, the incidence rate of anxiety was 51.8%, SAS score was (41.25±8.48), the incidence rate of depression was 30.4%, and the SDS score was (0.45±0.10) (P<0.05). Conclusions The health management of PCOS group practice can improve PCOS patients′negative emotions effectively.
8.Influence of infection frequency and vaccination on virus mutation of SARS-CoV-2
Guo XU ; Huan FAN ; Jianguang FU ; Huiyan YU ; Fei DENG ; Zhuhan DONG ; Shihan ZHANG ; Fengcai ZHU ; Changjun BAO ; Liguo ZHU
Chinese Journal of Experimental and Clinical Virology 2024;38(5):481-488
Objective:To analyze the effects of SARS-CoV-2 infection and vaccination on virus mutation.Methods:The whole genome sequencing sequences of 2 659 local SARS-CoV-2 specimens from Jiangsu Province in 2023 were selected for analysis, and relevant information such as demographic and clinical characteristics were collected, and the effects of infection and vaccination on the genome-wide mutation rate and S gene′s selective pressure of the virus were analyzed by univariate and multivariate linear regression models.Results:The average age of these infected patients was 55.0 (31.0, 74.0) years, 1 150 cases (43.2%) in the age group of ≥60 years, 1 367 cases (51.4%) were males, 2 044 cases (76.9%) had a history of COVID-19 vaccination, and 1 629 cases (61.3%) had the first-time infection. The clinical symptoms of the infected patients were mainly mild, with a total of 2434 cases (91.5%), and 29 cases (1.1%) with severe symptoms or more. The average substitution rate of SARS-CoV-2 was 9.69 (9.38, 9.98)×10 -4 subs/site/year, and the dN/dS value of the S gene was 6.08 (5.56, 8.66), which was significantly greater than that of 1 ( P<0.001), indicating positive selection. The result of univariate and multivariate linear regression model analysis showed that the SARS-CoV-2 substitution rate was higher in those with vaccination history and reinfection, aged 20-30 years, ≥60 years, and the SARS-CoV-2 substitution rate was lower in males with moderate clinical symptoms and severe disease and above. Those with a history of vaccination and reinfection, aged 50-60 years old, ≥60 years old have smaller S gene dN/dS. Conclusions:Under the immune pressure exerted by vaccination and infection, the genome-wide mutation of SARS-COV-2 accelerated, but the non-synonymous mutation rate of the S gene decreased. The mechanism causing these phenomena needs further study.