1.Influences of ultraviolet A (UVA) on the secretion and expression of chemokine CXCL11/I-TAC by HaCaT cells
Shiling GENG ; Shijun SHAN ; Tongwei ZHANG ; Jian WU ; Zhihua WANG ; Ting XIAO ; Chundi HE ; Hongduo CHEN
Chinese Journal of Dermatology 2010;43(3):181-183
Objective To investigate the influences of UVA on the secretion and expression of chemokine CXCL11/I-TAC by HaCaT cells induced by interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α). Methods HaCaT cells were cultured in the presence of IFN-7 and TNF-a and irradiated with UVA of 2, 4 and 8 J/cm~2, respectively; those cells receiving neither treatment with IFN-γ or TNF-α nor UVA irradiation served as the negative control, and those receiving only cytokine treatment but no irradiation as the positive control. After another 24-hour culture, enzyme-linked immunosorbent assay (ELISA) was performed to detect the protein levels of CXCL11/I-TAC in the supernatant of HaCaT celb, real time PCR to measure the mRNA expression of CXCL11/I-TAC in these HaCaT cells. Results As far as the negative control HaCaT cells were concerned, there was a minor secretion of CXCL11/I-TAC protein and expression of CXCL11/I-TAC mRNA. After treatment with IFN-7 and TNF-a of 10 μg/L, the protein and mRNA expressions of CXCL11/ I-TAC were synergistically upregulated, whereas the induced secretion and expression of CXCL11/I-TAC by HaCaT cells were dose-dependently inhibited by UVA irradiation. Conclusions UVA irradiation inhibits the secretion and expression of CXCL11/I-TAC by HaCaT cells, which in turn suppresses the chemotaxis of Th1/ Tel cells in some degree.
2.Comparison of transfection efficiency of two kinds of cells by lipofection and electroporation
Hexuan ZHANG ; Keren SHAN ; Yan HE ; Ting ZHANG ; Chanjuan WANG ; Zhizhong GUAN
Chongqing Medicine 2014;(33):4432-4433
Objective To compare the transfection efficiency between different transfection methods in human HepG2 and SGC7901/ADM cells so as to provide experimental basis for further study .Methods To electrons fect the enhanced GFP plasmid into HepG2 and SGC7901/ADM cells by lipofection and electroporation methods ,respectively .The survival rates and transfection efficiency were analyzed .Results The efficiency of eGFP vector transfected into HepG2 cells by lipofection was (23 .8 ± 2 .1)% , compared with lipofection method ,the efficiency of eGFP plasmid transfected by electroporation was up to (49 .6 ± 2 .5)% ,and the difference was statistically significant(P<0 .05) .The efficiency of SGC7901/ADM cells by lipofection was (25 .4 ± 1 .3)% ,com‐pared with lipofection method ,the efficiency of electroporation was up to(52 .6 ± 2 .1)% ,and the difference was statistically signifi‐cant(P<0 .05) .This study provides reliable test parameters for electransfection of HepG2 and SGC7901/ADM cells .Conclusion The transfection efficiency of large fragment vector is efficiently improved by electroporation .
3.The clinical application of Mycoplasma pneumoniae antibody in children′s respiratory tract infection
Xiujuan PENG ; Ting XIANG ; Yongneng MA ; Fang HE ; Zili YANG ; Shan LIU ; Haoyue REN
International Journal of Laboratory Medicine 2017;38(17):2353-2354
Objective To detect mycoplasma pneumoniae antibody in children having the upper respiratory tract infection.And then investigate mycoplasma pneumoniae infection status of different season different age children.Methods In 5 403 cases of suspected pneumonia mycoplasma infection of 0 to 14 years old children using the method of passive particle agglutination determination of mycoplasma pneumoniae antibody,and analysis of the statistical results.Results The positive rate was 67.8% in the groups of children.The rates of infection was biggest during 2 to 3 years old children and 3-4 years old children,14.9% and 18.4%,respectively.In addition,we found that the highest rate of mycoplasma pneumoniae infection arised from October to January every year of the following year.Conclusion The infection of mycoplasma pneumoniae is on the rise,and children aged 0 to 6 years old are the main population.
4.Correlation between fibroblast growth factor receptor 2 polymorphisms and endemic fluorosis
Mei LIU ; Yan HE ; Jie DENG ; Ting ZHANG ; Chanjuan WANG ; Keren SHAN ; Zhizhong GUAN
Chinese Journal of Endemiology 2015;34(8):559-563
Objective To investigate the correlation between fibroblast growth factor receptor 2 (FGFR2) gene polymorphism and endemic fluorosis.Methods In Bijie City,Guizhou Province coal-burning-borne high fluoride areas,148 patients with fluorosis were selected as endemic fluorosis group;in non high fluoride areas of Changshun County of Guizhou Province,134 healthy people were selected as control group.Short tandem repeats (STRs)-PCR was utilized to detected the FGFR2 rs35668561 and D10S14839 microsatellite polymorphisms in endemic fluorosis cases and controls.Results FGFR2 rs35668561 461 bp (22AG)allele frequency of endemic fluorosis group (1.01%) was significantly lower than that of the control group (3.36%,x2 =5.29,P < 0.05).FGFR2 D10S14839 286 bp (9GT),300 bp (16GT),310 bp (21GT) and 314 bp (23GT) allele frequency in the endemic fluorosis group were 14.53%,11.82%,16.89% and 8.11%,in the control group were 22.01%,6.34%,8.96% and 16.42%,the difference was statistically significant.Then 300 bp (16GT)and 310 bp (21GT)allele frequency of endemic fluorosis group was significantly higher than that of the control group (x2 =6.82,7.77,all P < 0.05),and 286 bp (9GT),314 bp (23GT) allele frequency of endemic fluorosis group was significantly lower than that of the control group (x2 =5.32,9.16,all P < 0.05).Conclusions FGFR2 rs35668561 and D10S14839 polymorphism are associated with endemic fluorosis.FGFR2 rs35668561 461 bp (22AG) allele may be a protective factor of endemic fluorosis.D10S14839 300 bp (16GT) and 310 bp (21GT) allele may be risk factors of endemic fluorosis,286 bp (9GT) and 314 bp (23GT) allele may be protective factors of endemic fluorosis.
5.A study of mRNA expressi on and transcription regulation in the promoter region of myeloperoxidase gene from a population living in the area with coal-burning endemic fluorosis in Guizhou Province
Xi TU ; Keren SHAN ; Yan HE ; Dong AN ; Shiqing XU ; Shuguang ZHOU ; Ting ZHANG ; Changxue WU ; Yi LI ; Zhizhong GUAN
Chinese Journal of Endemiology 2014;(4):374-378
Objective To explore the correlation between myeloperoxidase (MPO) genetic variation and coal-burning endemic fluorosis, and to understand the influence of integrated intervention including stove changes and health education on people’s health in the area. Methods In 2007, coal-burning endemic fluorosis disease areas were selected in Bijie City, Guizhou Province. No stove changes in Yachi Town, 150 patients with dental fluorosis were selected as fluorosis non-intervention group, and the intervention group was 150 patients in Changchun Town where the stoves were changed 2 years ago. The population in control group was selected in an area with non-endemic fluorosis in Changshun County. The mRNA expressions of MPO in leukoxytes were detected by real-time PCR. HepG2 cells were cultured in vitro and divided into four groups: pGL3-A group, pGL3-G group, pGL3-Control group and pGL3-Basic group. pGL3-A and pGL3-G were recombinant plasmid, while pGL3-Basic as a blank control and pGL3-Control as a positive one. The internal reference plasmid pRL-TK co-transfected the HepG2 cells with pGL3-G, pGL3-A, pGL3-Basic and pGL3-Control, respectively. The influence of sudden change of MPO gene promoter on the gene transfection activity was evaluated by a dual luciferasereporter gene system. Results The expression level of MPO mRNA in peripheral blood leukocytes in non-intervention group(0.054 ± 0 . 003 ) were higher than control and intervention groups (0.019 ± 0.004,0.019 ± 0.003, all P<0.05), and no significant change was found between intervention group and control group(P>0.05). After the MPO-463G/A locus genetic variation occured, the luciferase reporter gene expression level of the recombinant plasmid pGL3-G(0.753 4 ± 0.086 6) was higher than that of the pGL3-A(0.490 0 ± 0.022 3, P < 0.05). Conclusions The study on MPO gene promoter-463G/A locus has prompted that MPO gene allele may be a protective factor to coal-burning fluorosis. The integrated interventions have a role in the prevention and treatment of endemic fluorosis.
6.The Association of FGFR2 rs1219648 Polymorphism with Susceptibility of Breast Cancer in Han Population in Guizhou Province
Mei LIU ; Keren SHAN ; Yan HE ; Ting ZHANG ; Yan XIAO ; Changxue WU ; Chanjuan WANG ; Xiaoliang WANG ; Zhizhong GUAN ; Xilin REN
Chinese Journal of Clinical Oncology 2010;37(1):29-31
Objective:To investigate the association between polymorphisms of FGFR2 and the susceptibility of breast cancer in Han population in Guizhou province.Methods:Genotyping was performed using PCR-sequence-specific primers(PCR-SSP)in 106 histologically confirmed breast cancer cases and 116 cancer-free controls.Results:The genotype frequencies of rs1219648 TT,TC,and CC were 50%,25.47%.and 24.53% in breast cancer cases and 29.31%,48.28%,and 22.41% in the controls.The gene frequencies of T in breast cancer cases and the controls were 62.74% and 53.45%.respectively.The gene frequencies of C were 37.26% and 46.55%.respectively.The distribution of allele and genotype frequencies of FGFR2 rs1219648 was statistically different between breast cancer cases and the controls(P<0.05).Conclusion:FGFR2 rs1219648 polymorphism influences the susceptibility of breast cancer.TT genotype might serve as a risk factor for breast cancer.
7.Analysis of the GSTP1 gene Ile105Val polymorphism in fluorosis areas in Guizhou Province
Ting, ZHANG ; Ke-ren, SHAN ; Dong, AN ; Shi-qing, XU ; Shu-guang, ZHOU ; Yan, HE ; Chang-xue, WU ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2009;28(3):268-271
Objective To investigate plasma glutathione S-transferase(GSTs) activity and GSTP1 gene Ile105Val polymorphism in Bijie City, Guizhou Province, a coal-burning fluorosis endemic area. Methods One hundred and sixty villagers from Yachi Twon using non-improved cooking stoves were selected as the non-intervened group in Bijie City, Guizhou Province where coal-burning fluorosis was prevailing; 153 villagers as the intervented group were chosen from Changchun Twon, where cooking stoves were improved; 151 villagers were served as the control group from Baiyunshan Twon, Changshun County without endemic fluorosis. The activity of GSTs was tested by colorimetric analysis with spectrophotometer. The genotype of the GSTP1 gene Ile105Val polymorphism, presenting as either homozygous wild-type (AA), or heterozygous mutation type (AG), or homozygous mutation type (GG), was detected through the PCR-RFLP procedure. Results The activity of GSTs in plasma of non-intervened group [(12.44±4.97) kU/L]was significantly lower than that of intervened group (P < 0.05), and that of intervened group[(20.78±6.20)kU/L]was significantly lower than that of control group[(24.30±6.27)kU/L, P< 0.05]. The difference of the enzyme activity of three groups were statistically significant (F = 51.71, P < 0.05), but this enzyme activity did not vary significantly in each sex of each grnup(P > 0.05). Compared intervened group [AA:67.3%(103/153), AG:29.4%(45/153),GG:3.3%(5/153)]and non-intervened group[AA:66.9%(107/160), AG:30%(48/160), GG:3.1%(5/160)]with control group[AA:74.8%(113/151), AG:25.2%(38/151), GG:0 (0/151)], the Ile105Val polymorphism site of GSTP1 gene had significant difference(χ2= 6.04,6.07, both P< 0.05), but not significant between intervened and non-intervened groups(χ2 = 0.02, P>0.05). Conclusions Fluorosis can decrease the activity of GSTs and introduce the GSTP1 gene Ile105Val polymorphism, intervention with the fluorine intake will improve the effect of fluoride on the body.
8.Expression of USP15, TβR-I and Smad7 in psoriasis.
Ai-Ping, FENG ; Yi-Min, HE ; Xin-Xin, LIU ; Jia-Wen, LI ; Ya-Ting, TU ; Feng, HU ; Shan-Juan, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(3):415-9
The deubiquitinating enzyme ubiquitin specific peptidase 15 (USP15) is regarded as a regulator of TGFβ signaling pathway. This process depends on Smad7, the inhibitory factor of the TGFβ signal, and type I TGFβ receptor (TβR-I), one of the receptors of TGFβ. The expression level of USP15 seems to play vital roles in the pathogenesis of many neoplasms, but so far there has been no report about USP15 in psoriasis. In this study, immunohistochemical staining of USP15, TβR-I and Smad7 was performed in 30 paraffin-embedded psoriasis specimens and 10 normal specimens to investigate the expression of USP15, TβR-I and Smad7 in psoriasis and to explore the relevance among them. And USP15 small interfering RNA (USP15 siRNA) was used to transfect Hacat cells to detect the mRNA expression of TβR-I and Smad7. Of 30 cases of psoriasis in active stage, 28, 24 and 26 cases were positive for USP15, TβR-I and Smad7 staining, respectively. The positive rates of USP15 and Smad7 were significantly higher in psoriasis specimens than in normal skin specimens (44.1%±26.0% vs. 6.1%±6.6%, 47.2%±27.1% vs. 6.6%±7.1%), and positive rate of TβR-I (20.3%±22.2%) in psoriasis was lower than that in normal skin specimens (46.7%±18.2%). There was a significant positive correlation between USP15 and Smad7 expression, and significant negative correlations between USP15 and TβR-expression, an I d between TβR- and Smad7 expression I in psoriasis. After transfection of USP15 siRNA in Hacat cells, the expression of TβR-mRNA was up I -regulated and that of Smad7 was down-regulated. It is concluded that USP15 may play a role in the pathogenesis of psoriasis through regulating the TβR-I/Smad7 pathway and there may be other cell signaling pathways interacting with USP15 to take part in the development of psoriasis.
9.Role of VEGF-D in lymphangiogenesis of breast cancer and relationship with prognosis
xiao-liang, QI ; yan, GU ; rui, ZHANG ; shan-yu, GUO ; yong-feng, XIA ; liu, HE ; ting-ting, TANG ; yong-yang, BAO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(11):-
Objective To evaluate the role of vascular endothelial growth factor-D(VEGF-D)in lymphangiogenesis of breast cancer,and investigate its relationship with some clinicopathological parameters and prognosis. Methods VEGF-D expression was detected in 85 cases with primary breast carcinoma by immunohistochemistry,and VEGF-D mRNA was detected by RT-PCR.Podoplanin monoclonal antibody was used to mark lymphatic endothelial cell and lymphatic vessel density(LVD) was counted.The relationship among the aboved parameters,clinicopathological parameters and prognosis was analysed. Results It was revealed by immunohistochemistry that VEGF-D expression was ranked by 5 levels: negative,n=5(5.88%);"+",n=17(20%);"++",n=34(40%);"+++",n=22(25.88%),and "++++",n=7(8.24%).VEGF-D expression was associated with tumor lymph node metastasis and TNM clinical stage(P
10.A study of interleukin-10 gene polymorphisms in Miao, Dong and Buyi ethnics of Guizhou.
Chan-juan WANG ; Ke-ren SHAN ; Yan HE ; Ting ZHANG ; Yi LI ; Chang-xue WU ; Chan ZHANG ; Zhi-zhong GUAN
Chinese Journal of Medical Genetics 2013;30(1):116-120
OBJECTIVETo investigate allelic frequencies of interluekin-10 (IL-10) gene promoter in Miao, Dong and Buyi ethnics of Guizhou.
METHODSTaqMan MGB-based real-time PCR was used to determine the genotypes of IL-10 -819 and IL-10 -592 in 589 Miao, Dong and Buyi ethnics of Guizhou.
RESULTSThe allelic frequency of IL-10 -819 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. Allelic frequencies of IL-10 -592 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. In Miao, Dong and Buyi ethnics, the distributions of genotype frequencies of IL-10 -819 and IL-10 -592 were statistically different from Han ethnics from Guizhou and Taiwan of China as well as South Koreans.
CONCLUSIONThere is a heterogeneity in the frequencies of polymorphisms of IL-10 promoter among different ethnic groups.
Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Interleukin-10 ; genetics ; Polymorphism, Single Nucleotide ; Population Groups ; genetics ; Promoter Regions, Genetic