1.Role of NO Pathway in Membrane Estrogen Receptor Mediated Proliferation and Apoptosis of Endothelial Progenitor Cells
Zhi TAN ; Yuhong CUI ; Qiuling XIANG ; Guiping LIN ; Tinghuai WANG
Journal of Sun Yat-sen University(Medical Sciences) 2010;31(1):64-68
[Objective] The aim of the present study was to investigate the role of membrane estrogen receptor (mER) mediated pathway in the proliferation and apoptosis of endothelial progenitor cells (EPCs). [Methods] Bone marrow (BM)-derived EPCs were cultured. The cells were divided into different groups, plus or not plus estrogen receptor blocker (ICI 182,780), PI3K inhibitors (LY294002), and NOS inhibitor (L-NAME) to show the effect of E_2-BSA on EPCs. The proliferation of EPCs was determined by MTT and nitric oxide (NO) release was measured by chromatometry. Apoptotic cell death was determined using the Hochest 33258 staining. The expression of phosphorylated eNOS (p-eNOS) were detected by Western blot. [Results] E_2-BSA could increase EPCs proliferation, and this effect was inhibited by estrogen receptor blocker ICI 182,780, thus indicated that mER-initiated membrane signaling pathways were involved in the action of estrogen on EPCs. E_2-BSA increased nitric oxide production and inhibited apoptosis induced by serum withdrawal, and this effect also inhibited by PI3K inhibitor (LY294002), NOS inhibitor (L-NAME)and estrogen receptor blocker(ICI 182,780), thus indicated that PI3K/Akt/NO pathway was involved the effect of estrogen on EPCs apoptosis. Moreover, E_2-BSA treatment increased phosphorylation of eNOS (p-eNOS). PI3K inhibitors (LY294002) also blocked these effects. [Conclusions] The results of present study suggested that mER mediated EPCs proliferation and apoptosis were related to the PI3K/Akt/eNOS pathway.
3.HLA-DQA1 genotyping by PCR-SSP technique in Jing nationality of Central Vietnam
Ting-Ping CHEN ; Lin-Lin WANG ; Wei-Xiong LIN ; Jian-Feng CHEN ; Xiang-Zhi XIE ;
Chinese Journal of Immunology 1985;0(03):-
Objective:To investigate the polymorphism of HLA-DQA1 genes in Jing nationality of Central Vietnam.Methods:Applied PCR-SSP tecnique to determine the polymorphism of the HLA-DQA1 alleles of 105 healthy children and youth,unrelated individuals in Central of Vietnam.Results:10 HLA-DQA1 alleles were detected of which DQA1*0104 were the most common allele with frequency of 21.3% and lowest frequency is DQA1*0601.Conclusion:The results indicate that HLA-DQA1 alleles polymorphism of Jing nationality in Central Vietnam is different from the other Chinese. [
4.Relationship between Gene Polymorphisms in Interleukin-12B Promoter and Upper Gastrointestinal Disease Infected with Helicobacter Pylori in Children and Adolescent
qing-wen, SHAN ; cheng-xue, JING ; lin-lin, WANG ; zi-li, LV ; xiang-zhi, XIE ; qing, TANG ; xiang, YUN
Journal of Applied Clinical Pediatrics 2006;0(19):-
Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.
5.Exploration on the Research-oriented Teaching Reform of Microbiology
Yue-Lan YIN ; Xin-An JIAO ; Zhi-Ming PAN ; Lin SUN ; Jin-Lin HUANG ; Xiang CHEN ;
Microbiology 2008;0(12):-
This paper is discussed about course system construction of Microbiology, teaching method, in- struction means and experimental teaching mode. Teaching practice indicated that reform the pattern of Mi- crobiology educational mode can stimulate students’ interest in studying the course, cultivate their inde- pendent ability to solve questions, develop their creative thinking. It is an important way to train high-caliber talents.
6.Design, synthesis and antiproliferative activities of artemisinin derivatives substituted by N-heterocycles.
Zhi-zhong ZUO ; Hang ZHONG ; Ting CAI ; Yu BAO ; Zhi-qiang LIU ; Dan LIU ; Lin-xiang ZHAO
Acta Pharmaceutica Sinica 2015;50(7):868-874
Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.
Antineoplastic Agents
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chemical synthesis
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chemistry
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Artemisinins
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chemical synthesis
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chemistry
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Breast Neoplasms
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pathology
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Cell Proliferation
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Doxorubicin
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Drug Design
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HL-60 Cells
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drug effects
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Humans
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MCF-7 Cells
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drug effects
7.Enhancement of artemisinin biosynthesis in transgenic Artemisia annua L. by overexpressed HDR and ADS genes.
Ya-Xiong WANG ; Shi-Ping LONG ; Li-Xia ZENG ; Li-En XIANG ; Zhi LIN ; Min CHEN ; Zhi-Hua LIAO
Acta Pharmaceutica Sinica 2014;49(9):1346-1352
Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua
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genetics
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metabolism
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Artemisinins
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metabolism
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Biosynthetic Pathways
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Drugs, Chinese Herbal
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Mixed Function Oxygenases
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genetics
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Oxidoreductases
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genetics
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Plant Proteins
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genetics
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Plants, Genetically Modified
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genetics
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metabolism
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Plants, Medicinal
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genetics
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metabolism
8.Sinus histiocytosis with massive lymphadenopathy in infant: report of a case.
Hui-yun LIN ; Lan-xiang GAO ; Guang LIU ; Guang-zhi YANG
Chinese Journal of Pathology 2009;38(9):630-631
Diagnosis, Differential
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Histiocytoma, Benign Fibrous
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metabolism
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pathology
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Histiocytosis, Langerhans-Cell
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metabolism
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pathology
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Histiocytosis, Sinus
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metabolism
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pathology
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surgery
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Humans
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Infant
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Male
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S100 Proteins
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metabolism
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Xanthogranuloma, Juvenile
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metabolism
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pathology
9.Enhancement of artemisinin biosynthesis in transgenic Artemisia annua L. by overexpressed HDR and ADS genes.
Yaxiong WANG ; Shiping LONG ; Lixia ZENG ; Lien XIANG ; Zhi LIN ; Min CHEN ; Zhihua LIAO
Acta Pharmaceutica Sinica 2014;49(9):1346-52
Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
10.Influence and mechanism of He-Ne laser on scar formation of filtration canal after trabeculectomy in rabbit
Rui-Lin, WANG ; Gui-Gang, LI ; Zhi-Qi, CHEN ; Yan, XIANG ; Hong, ZHANG
International Eye Science 2010;10(9):1650-1653
AIM: To investigate the influence of He-Ne laser on connective tissue growth factor (CTGF) expression and collagen formation of fibroblast in filtration site after trabeculectomy in rabbit, and to discuss the mechanism for preventing scar formation with He-Ne laserin vivo.METHODS: The upper nasal limbus area next to the upper rectus muscle in right eyes Received 10 minutes He-Ne laser irradiation (200mW/cm2) every day for three days, the left eyes served as control. Twenty-four hours after the last irradiation, both eyes of the rabbits were took trabeculectomy surgery. The expressions of CTGF in the filtration area were tested on the 7th, 14th and 28th day after surgery and collagen density was tested on the 14th and 28th day after surgery. Each of the time point had 7 rabbits. RESULTS: The expression of CTGF was lower than that of the control group's on the 7th and 14th day after trabeculectomy surgery (P=0.01, P=0.005). When examined on the 14th and 28th day, the collagen density of irradiation group were significantly lower than that of the control group's (P=0.013, P=0.01).CONCLUSION: Pretreating the filtration area with 200mW/cm2 He-Ne laser may be helpful in preventing scar formation after trabeculectomy in rabbit, possibly due to downregulation of the expression of CTGF and collagen synthesis in fibroblasts. He-Ne laser may be developed into a new scar preventing method in filtration surgery.