1.Expression of glutathione S-transferase mRNA in peripheral blood of the population in coal-burning fluorosis area
Chang-xue, WU ; Ke-ren, SHAN ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2010;29(2):121-123
Objective To investigate expression of glutathione S-transferase(GST) mRNA in peripheral blood of the population in coal-burning fluorosis area and to evaluate the effect of comprehensive control intervention. Methods Fifty samples of peripheral blood from patients in the coal-buring fluorosis area in Bijie county of Guizhou province were selected as fluorasis group and 50 samples of peripheral blood from patients in area with comprehensive management were selected as intervention group, respectively. Fifty samples from non-endemic fluorosis area were selected as the control group. Total RNA from blood was extracted and purified by the Trizol- Phenol-Chloroform one-step method. Expression of GST mRNA was detected by using SYBR Green I real-time fluorescence quantitative PCR. Results The data of GST mRNA in fluorosis group, intervention group and control group was 38.28±27.22,70.56±37.23 and 103.46 ± 46.62, respectively. There was a significant difference between the groups(F = 3.75, P < 0.05). Decreased expression of GST mRNA in fluorosis group and intervention group as compare to control was detected(all P < 0.05), and the expression of GST mRNA in intervention group was higher than that in fluorosis group(P < 0.05). Conclusion Coal-burning fluorosis possibly led to the decreased expression of GST mRNA in peripheral blood, and comprehensive control maybe prevent the decreased expression of GST in mRNA level.
2.Evaluation of a health education project on endemic fluorosis in Shandong Province in 2010
yu-xue, WEN ; Zhong-jie, YUN ; Shan-shan, WEN ; Wei-ping, SHEN
Chinese Journal of Endemiology 2013;32(5):576-579
Objective To evaluate the effects of a health education project on endemic fluorosis in Shandong Province,and to provide a basis for formulating control strategies.Methods From December 2010 to June 2011,according to historical conditions,a total of 19 counties (cities,districts) of Shandong Province were chosen,and 3 townships (towns) were chosen in each project county.Health educational activities on endemic fluorosis were carried out in the Central Primary School in grade 4 to 6 in each township(town).In each project township(town),3 villages were chosen in each selected township(town) where the health educational activities in the community were carried out.Before and after the health educational activities,surveys on knowledge questionnaire on drinking-water-borne fluorosis control were conducted among 30 students of grade 5 in the Central Primary School and 15 housewives in every school location in each selected township(town).Results After the health educational activities,the knowledge awareness rates of endemic fluorosis control of the students and housewives were 96.53% (5482/5679) and 94.88% (3501/3690),respectively,and increased significantly compared with those before intervention [62.31% (5154/8271) and 76.91% (2815/3660)],and the difference was statistically significant (x2 =2176.50,490.58,all P < 0.01).Among the primary school students and housewives,the knowledge awareness rates of endemic fluorosis control were increased by 34.22% and 17.97%,respectively.Conclusions Health education activities on endemic fluorosis can significantly improve the knowledge awareness of target population,which will play a positive role in promoting prevention and control of endemic fluorosis.
3.Long noncoding RNA LINC00520 prevents the progression of cutaneous squamous cell carcinoma through the inactivation of the PI3K/Akt signaling pathway by downregulating EGFR.
Chinese Medical Journal 2019;132(4):454-465
BACKGROUND:
Long noncoding RNAs (lncRNAs) play pivotal roles in various malignant tumors. Epidermal growth factor receptor (EGFR) signaling is associated with the pathogenesis of cutaneous squamous cell carcinoma (cSCC). This study aimed to explore the role of LINC00520 in the development of cSCC via EGFR and phosphoinositide 3-kinase-protein kinase B (PI3K/Akt) signaling pathways.
METHODS:
A microarray analysis was applied to screen differentially expressed lncRNAs in cSCC samples. The A431 cSCC cell line was transfected and assigned different groups. The expression patterns of LINC00520, EGFR, and intermediates in the PI3K/Akt pathway were characterized using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting analysis. Cell proliferation, migration, and invasion were detected using the MTT assay, scratch test, and Transwell assay, respectively. Cell-based experiments and a tumorigenicity assay were conducted to assess the effect of LINC00520 on cSCC progression. This study was ended in September 2017. Comparisons between two groups were analyzed with t-test and comparisons among multiple groups were analyzed using one-way analysis of variance. The nonparametric Wilcoxon rank sum test was used to analyze skewed data. The enumerated data were analyzed using the chi-square test or Fisher exact test.
RESULTS:
Data from chip GSE66359 revealed depletion of LINC00520 in cSCC. Cells transfected with LINC00520 vector and LINC00520 vector + si-EGFR showed elevated LINC00520 level but decreased levels of the EGFR, PI3K, AKT, VEGF, MMP-2 and MMP-9 mRNAs and proteins, and inhibition of the growth, migration and adhesion of cSCC cells, while the si-LINC00520 group showed opposite trends (all P < 0.05). Compared with the LINC00520 vector group, the LINC00520 vector + si-EGFR group showed decreased levels of the EGFR, PI3K, AKT, VEGF, MMP-2 and MMP-9 mRNAs and proteins, and inhibition of the growth, migration and adhesion of cSCC cells, while the LINC00520 vector + EGFR vector group showed opposite results (all P < 0.05).
CONCLUSION
Based on our results, LINC00520-targeted EGFR inhibition might result in the inactivation of the PI3K/Akt pathway, thus inhibiting cSCC development.
Animals
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Carcinoma, Squamous Cell
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pathology
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prevention & control
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Cell Line, Tumor
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Cell Movement
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Cell Proliferation
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Disease Progression
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ErbB Receptors
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antagonists & inhibitors
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Female
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Humans
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Lymphatic Metastasis
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Mice
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Neoplasm Invasiveness
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Phosphatidylinositol 3-Kinases
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physiology
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Proto-Oncogene Proteins c-akt
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physiology
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RNA, Long Noncoding
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physiology
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Signal Transduction
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physiology
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Skin Neoplasms
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pathology
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prevention & control
4.Histocompatibility and imprinting status of parthenogenetic embryonic stem cells.
Yuan XUE ; Zhiyan SHAN ; Zhong ZHENG ; Lei LEI
Journal of Biomedical Engineering 2010;27(5):1158-1161
The parthenogenetic embryonic stem cells (pESCs) derived from parthenogenetic embryos have the totipotency and proliferation capacity similar to those of the fertilized embryonic stem cells (fESCs). Therefore, the establishment of pESCs line avoids destroy of embryo and kence may make pESCs less concerns with political and ethical issues. These cells are characterized by their histocompatibility with the oocyte donor and therefore is more suitable for cell and tissue replacement therapy. In addition, because of the typical imprinting status, pESCs also provide a valuable in vitro model system for studying the molecular mechanisms in genomic imprinting.
Animals
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Embryonic Stem Cells
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cytology
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Female
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Gene Expression Profiling
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methods
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Gene Expression Regulation, Developmental
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genetics
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physiology
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Genomic Imprinting
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Histocompatibility
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Parthenogenesis
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genetics
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physiology
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Pluripotent Stem Cells
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cytology
5.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.
6.Modified shock index and mortality rate of emergency patients
Ye-Cheng LIU ; Ji-Hai LIU ; Amy-Zhe FANG ; Guang-Liang SHAN ; Jun XU ; Zhi-Wei QI ; Hua-Dong ZHU ; Zhong WANG ; Xue-Zhong YU
World Journal of Emergency Medicine 2012;3(2):114-117
BACKGROUND: This study aimed to determine whether modified shock index (MSI) is associated with mortality that is superior to heart rate, blood pressure, or the shock index (SI) in emergency patients.METHODS: A retrospective database review was performed on 22161 patients who presented to Peking Union Medical College Hospital Emergency Department and received intravenous fluids from January 1 to December 31, 2009. We gathered data of the patients on age, gender, vital signs, levels of consciousness, presenting complaints, and SI and MSI were calculated for all patients.RESULTS: Multivariate regression analysis was performed to determine the correlation between risk factors and outcome. There is a significant correlation between emergency patient mortality rate and patient's vital signs obtained at the triage desk (HR>120 beats/min, systolic BP<90 mmHg, diastolic BP<60 mmHg). MSI is a stronger predictor of emergency patient mortality compared to heart rate and blood pressure alone, whereas SI does not have a significant correlation with emergency patient mortality rate.CONCLUSION: MSI is a clinically significant predictor of mortality in emergency patients. It may be better than using heart rate and blood pressure alone. SI is not significantly correlated with the mortality rate of the emergency patient.
7.Effect of Ginkgo biloba extract preconditioning on discordant cardiac xenografts.
Xue-shan HUANG ; Xuan LIU ; Dao-zhong CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2006;26 Suppl():108-111
OBJECTIVETo investigate the effect of ginkgo biloba extract (ginaton) preconditioning on discordant cardiac xenografts from guinea pig to rat, and explore its mechanism.
METHODSCervical cardiac transplantation model was established in the rats,which were divided into 4 groups Group 1 (cobra venom factor ( CVF) pretreatment, n = 10]; Group 2 (CVF + ginaton, n = 5) ; Group 3 Ccyclosporine (CsA); Group 4 (CVF + CsA + ginaton, n = 8]. The survival time and histopathology after xenograft were observed and expressions of intercellular adhesion molecule-1 (ICAM-1) heme oxygenase-1 (HO-1) CD68 and CD57 were detected.
RESULTSPathologic manifestion of grafts showed changes of acute vascular rejection (AVR) in all groups. The mean survival time after car diac xenograft was 41 hrs in Group 1, 68 hrs in Group 2, 55 hrs in Group 3 and 74 hrs in Group 4. Expression of intercellular adhesion molecule-1 (ICAM-1 ) decreased after ginaton preconditioning (P < 0. 05). CD68 and CD57 expressions were down-regulated, HO-1 expression was up-regulated, as well as the apoptotic index (Al) reduced significantly after ginaton with cyclosporine A preconditioning.
CONCLUSIONGinaton preconditioning can prolong the survival time after discordant xenograft, and significantly alleviate pathological lesion from acute xenograft vascular rejection combined with cyclosporine A.
Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; CD57 Antigens ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Ginkgo biloba ; Guinea Pigs ; Heart ; drug effects ; Heart Transplantation ; Heme Oxygenase-1 ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Myocardium ; immunology ; metabolism ; Rats ; Transplantation Conditioning ; Transplantation, Heterologous
8.Inhibitory effects of 17beta-estradiol on spontaneous and activated contraction of rat uterus smooth muscle.
Qin MA ; Hong-Fang LI ; Shan JIN ; Xing-Cheng DOU ; Ying-Fu ZHANG ; Li-Xue ZHANG ; Zhong-Rui DU
Chinese Journal of Applied Physiology 2013;29(4):305-309
OBJECTIVETo observe and compare the effects of 17beta-estradiol (EST) on the phasic and tonic contractile activities of the uterine smooth muscles of SD rats in vitro.
METHODSDifferent concentrations of 17beta-estradiol were added into the perfusion muscular sockets containing uterine smooth muscles of SD rats, and the activities of muscle contraction were recorded at the same time.
RESULTS17beta-estradiol had obvious depression effects on spontaneous rhythmic contraction of the uterine smooth muscles in a concentration-dependent manner, it could considerably decrease muscular tension, the mean amplitudes and frequencies of contractile waves (P < 0.01); it could also suppress the uterine contraction stimulated by KCl, CaCl2 or prostaglandin F2alpha (PGF2alpha). Based on the contraction of uterine smooth muscle stimulated by KCl, IC50 was 7.278 micromol/L and pD2 was -0.862 when calculated by linear regression method. 17beta-estradiol could also inhibit the maximal CaC12 contraction of uterine smooth muscle in the Ca2+ free Krebs solution, which the ECQ was 1.422 x 10(-3) mol/L, pD2 was 2.847 (control), but the E50 was 3.028 x 10(-3) mol/L, p2 was 2.519 (added with EST) when calculated by linear regression method.
CONCLUSIONThe depression effects of 17beta-estradiol on the spontaneous rhythmic contraction and activated contraction of the uterine smooth muscles of SD rats could be mediated through the blockage of C2+ influx through potential-dependent Ca2+ channels of plasma membrane.
Animals ; Estradiol ; pharmacology ; Female ; Muscle, Smooth ; drug effects ; Myometrium ; drug effects ; Rats ; Rats, Sprague-Dawley ; Uterine Contraction ; drug effects
9.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
10.Construction and functional characterization of a monocistronic replicon based on the HCV genotype 2a promotor.
Xue-Li LI ; Yu LEI ; Shan ZHONG ; Feng-Ying PENG ; Zhi ZHOU ; Kui LI ; Hong REN
Chinese Journal of Hepatology 2012;20(2):103-107
To construct a hepatitis C virus (HCV) genotype 2a monocistronic replicon and investigate its replication capabilities in the human hepatocarcinoma cell lines, Huh7.5 and Huh7.1, in order to determine its potential as a molecular tool for future in vitro studies of HCV replication and selection studies for putative anti-HCV drugs. Site-directed mutagenesis was used to delete the Core-E1-E2-p7-NS2 fragment (about 3090 bp) from plasmid pJ6JFH1BlaRL. The resultant trianglepJ6JFH1BlaRL plasmid was digested with AgeI and AvrII to release the cDNA fragment (hereafter, referred to as fragment L) containing partial 5'-untranslated region (UTR), the first 12 amino acid (aa) of HCV Core coding sequence, full-length coding sequences for the blasticidin-resistance gene, Renilla luciferase, foot-and-mouth disease virus (FMDV) 2a antiprotease and ubiquitin, and partial coding sequence for HCV NS3. To generate the monocistronic replicon, pSGRmJFH1BlaRL, fragment L was ligated into the pSGR-JFH1 vector that had been digested with AgeI and AvrII to remove the partial 5'-UTR, the first 19 aa of HCV Core coding sequence, the full-length coding sequence for the neomycin phosphotransferase II gene, the internal ribosomal entry site from encephalomyocarditis virus, and partial HCV NS3 coding sequence. A replication-defective mutant replicon, pSGRmJFH1BlaRL/GND, was constructed by a similar procedure using the pSGR-JFH1/GND vector. Fragment L was confirmed in both constructs by sequencing. Replicon RNAs were prepared from XbaI-linearized plasmid DNA templates with Invitrogen's T7 MEGAscript kit, and were purified by DNase I treatment and LiCl precipitation. RNAs were quanti?ed by optical density, and the quality and concentration were con?rmed by agarose gel electrophoresis. Replicon RNAs were transfected into Huh7.5 and Huh7.1 cells using Invitrogen's DMRIE-C transfection reagent at a ratio of 5 mug of lipid to 1mug of RNA. Time course assay of Renilla luciferase activity indicated the replicon's replication function. The pSGRmJFH1BlaRL monocistronic replicon and pSGRmJFH1BlaRL/GND replication-defective mutant replicon were successfully constructed. The pSGRmJFH1BlaRL replicon was replication-proficient in Huh7.5 and Huh7.1 cells, with replication peaking at 72 hours post-transfection and decreasing after 96 hours. No replication was detected at any time point post-transfection for the defective mutant replicon. A monocistronic replicon of HCV genotype 2a was constructed and shown to be replication-proficient in human hepatocarcinoma cell lines.
Cell Line, Tumor
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Genetic Vectors
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Genome, Viral
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Genotype
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Hepacivirus
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genetics
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Humans
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Mutagenesis, Site-Directed
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RNA, Viral
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Transfection
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Virus Replication