1.Involvement of nuclear factor of activated T-cells (NFATc) in calcineurin-mediated ischemic brain damage in vivo
Yingjun ZHANG ; Heshan MEI ; Chuan WANG ; Yongli WANG ; Yongjian ZHANG
Acta Pharmaceutica Sinica 2005;40(4):299-305
Aim To study the involvements of nuclear factor of activated T-cells (NFATc) and NFκB in calcineurin-mediated ischemic brain damage in vivo. Methods The rat transient forebrain ischemia conducted through 15 min ischemia followed by 8, 24, and 72 h reperfusion was induced using the fourvessel method. The rats were divided randomly into five groups; sham control group, ischemia/reperfusion (I/R) group, CsA treated groups (for 8, 24, and 72 h reperfusion). Western blotting was performed to detect changes of FasL, NFATc, I-κB-α, and phospho-I-κB-α protein expression, and gel shift assays for NFAT FasL-DNA binding activities. Results Western blotting showed that the expressions of both FasL and NFATc protein were significantly increased in the hippocanpus of rat subjected to transient forebrain ischemia in comparison with those of the sham control group, which were markedly reduced by CsA. The I-κB-α protein showed no changes in all groups, and phospho-I-κB-α protein was not observed in this study. Proximal and distal FasL promoter NFAT sites bind NFAT proteins from the hippocampal neurons subjected to transient forebrain ischemia, and DNA-binding activities increased significantly compared with those of the sham control group. CsA markedly inhibited these changes. Conclusion NFATc may be involved in calcineurin-mediated ischemic brain damage and transcription factor NF-κB may not be involved.
2.Inhibition of valdecoxib on the proliferation of human gastric cancer cells
Junxia LI ; Suwen SU ; Heshan MEI ; Yongli WANG ; Chuan WANG ;
Chinese Pharmacological Bulletin 1986;0(04):-
AIM To study the effect and mechanism of valdecoxib, a selective COX 2 inhibitor, on human gastric cancer BGC 823 cells. METHODS MTT assay and flow cytometry were used to observe the effect of valdecoxib on proliferation, apoptosis and the cell cycle distribution of BGC 823 cells. Laser confocal microscopy, transmission electron microscope and DNA fragmentation assay were further used to detect the apoptosis. The content of LDH was examined using LDH kit. RESULTS Valdecoxib in 25~400 ?mol?L -1 significantly inhibited the proliferation of BGC 823 cell in a time and dose dependent fashion, the inhibition rate of proliferation was 24 0%~92 0% after 72 h, and the rate of apoptosis was increased from (2 6?0 7)% to (7 6?1 5) %~(16 5?1 5)%. 100~400 ?mol?L -1 valdecoxib also decreased the proliferation index and the proportion of cells in the S phase, increased the proportion of cells in the G 0/G 1 phase, but had no effect on the proportion of cell in the G 2/M phase. CONCLUSION Valdecoxib inhibits human gastric cancer BGC 823 cells growth by inducing apoptosis and cell cycle arrest. The growth inhibitory effect of 400 ?mol?L -1 valdecoxib is also associated with cell necrosis.
3.The study of genome DNA methylation in rheumatoid arthritis
Xiao-Yan XU ; Mei-Mei WANG ; Chuan-Shi XIAO ; Xiao-Feng LI ; Lai-Yuan WANG ;
Chinese Journal of Rheumatology 2003;0(08):-
Objective To study the genome DNA methylation in rheumatoid arthirits(RA)and the re- lated factors of DNA methylation.Methods Twenty-first cases with RA and 20 controls were recruited to par- ticipate the study.Plasma Hcy,SAM,SAH,the MTHFR gene C677T polymorphism and the expression of LFA-1 in CD4~+T cells was measured in all patients and controls.Results①The SAM levels were lower sig- nificantly in RA groups than in controls.The SAH levels were higher significantly in RA groups than in con- trols.②There was significant inverse correlation between plasma Hcy level and SAM level(r=-0.932,P<0.01). There was significant positive correlation between plasma Hcy level and SAH level(r=0.924,P<0.01).③The expression of LFA-1 in CD4~+T cells was higher significantly in RA groups than in controls.There was a signif- icant positive correlation between LFA-1 expression level and Hcy level(r=0.557,P<0.01),a significant in- verse correlation between LFA-1 expression level and SAM level(r=-0.651,P<0.01).④The MTHFR gene mu- tation lead to dramatically increase of Hcy,SAH level and the expression of LFA-1 level in CD4~+T cells and genome DNA hypomethylation.Conclusion①Hypomethylation of genome DNA is found in most RA pa- tients.②The factors associated with genome DNA hypomethylation include MTHFR gene mutation and hyper- homocysteinemia.③The expression of LFA-1 in CD4~+ T cells is higer in RA groups than in controls,which re- lates to the DNA methylation level and the MTHFR gene C677T polymorphism.
4.Effects of strophanthidin on intracellular calcium concentration in ventricular myocytes of guinea pig
Suwen SU ; Yanfang XU ; Heshan MEI ; Yajuan QI ; Jingxiang YIN ; Chuan WANG ; Yongjian ZHANG ; Yongli WANG
Acta Pharmaceutica Sinica 2008;43(3):259-266
Effect of strophanthidin (Str) on intracellular calcium concentration ([Ca2+]i) was investigated on isolated ventricular myocytes of guinea pig. Single ventricular myocytes were obtained by enzymatic dissociation technique. Fluorescent signal of [Ca2+]i was detected with confocal microscopy after incubation of cardiomycytes in Tyrode's solution with Fluo3-AM. The result showed that Str increased [Ca2+]i in a concentration-dependent manner. The ventricular myocytes began to round-up into a contracture state once the peak level of [Ca2+]i was achieved in the presence of Str (10 μmol·L-1), but remained no change in the presence of Str (1 and 100 nmol·L-1). Tetrodotoxin (TTX), nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str (1 and 100 nmol·L-1), but had no obvious effects on the action of Str (10 μmol·L-1). The elevation of [Ca2+]i caused by Str at all of the detected concentrations was partially antagonized by rynodine (10 μmol·L-1) or the removal of Ca2+ from Tyrode's solution. In Na+, K+-free Tyrode's solution, the response of cardiomycytes in [Ca2+]i elevation to Str (10 μmol·L-1) was attenuated, while remained no change to Str (1 and 100 nmol·L-1). TTX, nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str at all of the detected concentrations in Na+, K+-free Tyrode's solution. The study suggests that the elevation of [Ca2+]i by Str at the low (nomomolar) concentrations is partially mediated by the extracellular calcium influx through Ca2+ channel or a "slip mode conductance" of TTX sensitive Na+ channel. While the effect of Str at high (micromolar) concentrations was mainly due to the inhibition of Na+, K+-ATPase. Directly triggering the release of intracellular Ca2+ from sarcoplasmic reticulum (SR) by Str may be also involved in the mechanism of [Ca2+]i elevation.
5.Heterotopic osteogenesis in vivo of PHBHHx and human umbilical cord mesenchymal stem cells
Zhixu HE ; Mei WANG ; Chuan YE ; Yong WANG ; Xiao ZENG ; Junbiao ZHANG
Chongqing Medicine 2016;45(20):2740-2743,2747
Objective To explore the ability for constructing tissue engineering bone in vivo in complex scaffolds with PHB‐HHx as the scaffolds material and human umbilical cord mensenchymal stem cells (hUCMCs) as the seed cells .Methods hUCM‐SCs were inoculated into PHBHHx scaffolds to induce osteogenesis culture in vivo for two weeks ,the the induced group was the experimental group and those without instilling hUCMCs served as the control group ,the nude mouse subcutaneous implantation was performed .Then taking material at 1 ,3 ,5 months after implantation in vivo was performed for conducting HE ,collagenⅠim‐munohistochemical ,alkaline phosphatase staining and RT‐PCR .Results hUCMSCs showed good cellular adsorbability .The size and form in the experimental group basically maintained the original status ,and the osteogenesis specific indicators were positive ;but the control group did not keek the original status ,its volume was gradually shrunk until complete degradation ,and the osteogen‐esis specific indicators were negative .Conclusion The PHBHHx scaffolds combined with hUCMSCs has the capability of in vivo heterotopic constructing tissue engineering bone in nude mouse by in vitro osteogenic induction .
6.Effect of salidroside on regulating phenotypic expressionof human osteoblast-like MG-63 cells through HIF-1α signal passway
Lin QI ; Chuan WANG ; Guohua MEI ; Xiaochang WEN ; Liqun MAO ; Xin JIN ; Ling LI ; Yue WANG
Chinese Pharmacological Bulletin 2017;33(6):836-843
Aim To investigate the effect of salidroside on the phenotypic expression of osteoblast and the possible molecular mechanism in hypoxia environment.Methods MTT, Annexin V/PI double staining, alkaline phosphatase(ALP) activity assay and enzyme-linked immnosorbent assay(ELISA) were performed respectively to detect the effect of salidroside on regulating phenotypic expression of MG-63 cells exposed to hypoxia.Then RT-PCR and Western blot were conducted to detect the expression levels of Osterix and Runx2 which were related to differentiation.At last, we investigated the expression levels of components of the HIF-1α pathway by RT-PCR,Western blot and ELISA, respectively.Immunofluorescence confocal microscope technology and luciferase reporter assay were performed to explore nuclear translocation and transcriptional activity of HIF-1α.Results Salidroside could markedly promote MG-63 cell proliferation, inhibit hypoxia-induced apoptosis, stimulate cell differentiation and promote expression levels of components of the HIF-1α pathway in hypoxia environment.Conclusion Salidroside could regulate phenotypic expression of MG-63 cells through HIF-1α/VEGF signaling pathway in hypoxia environment.
7.In vitro biocompatibility of polyhydroxybutyrate-co-volerate membrane with human bone marrow mesenchymal stem cells
Tao ZHANG ; Zhixu HE ; Chuan YE ; Jielin LIU ; Minxian MA ; Bo SUN ; Mei WANG ; Hua YANG
Chinese Journal of Tissue Engineering Research 2014;(12):1920-1925
BACKGROUND:Polyhydroxybutyrate-co-volerate (PHBV) is a noticeable tissue engineering material of polyhydroxyalkanoates family. It has the properties of low immune rejection response and good biocompatibility, and its degradation products are non-toxic.
OBJECTIVE:To investigate the biocompatibility of PHBV membrane material and human bone marrow mesenchymal stem cel s in vitro.
METHODS:Human bone marrow mesenchymal stem cel s at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cel number of two groups at 1, 2 and 4 hours and got the cel adherent rate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cel proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cel s at days 3, 6, 9 and 12 in both groups. After cel s were seeded upon PHBV membrane for 5 days, the cel growth upon the material was examined under a scanning electron microscope.
RESULTS AND CONCLUSION:When the cel s were cultured for 1 hour, the adherent rate in the experimental group was lower than that in the control group;but there were no significant differences between two groups at the other two periods. No difference was found in the cel proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cel s seeded upon PHBV membrane for 5 days grew wel with spindle morphology and the intercel ular connections were tight and more extracel ular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cel s.
8.Angiomyolipoma of the kidney with lymph node involvement.
Chuan-Zhen WU ; Feng-Hua WANG ; Cheng-Mei LI ; Wen-Chang FANG ; Jia-Ni YU
Chinese Journal of Pathology 2005;34(11):715-715
Adrenalectomy
;
Adult
;
Angiomyolipoma
;
pathology
;
surgery
;
Follow-Up Studies
;
Humans
;
Kidney
;
pathology
;
Kidney Neoplasms
;
pathology
;
surgery
;
Lymph Node Excision
;
Lymph Nodes
;
pathology
;
Lymphatic Metastasis
;
Male
;
Nephrectomy
;
Ureter
;
surgery
9.Keratocytoma of submandibular gland:report of a case.
Ji-man LI ; Ying WANG ; Jin-chuan FAN ; Mei TANG ; Jun HOU ; Lun LI
Chinese Journal of Pathology 2013;42(12):838-839
Carcinoma, Squamous Cell
;
pathology
;
Child
;
Cysts
;
pathology
;
Diagnosis, Differential
;
Epithelium
;
pathology
;
Female
;
Follow-Up Studies
;
Humans
;
Keratin-19
;
metabolism
;
Keratin-5
;
metabolism
;
Keratin-6
;
metabolism
;
Keratin-7
;
metabolism
;
Keratins
;
metabolism
;
Neoplasm Recurrence, Local
;
surgery
;
Reoperation
;
Submandibular Gland
;
surgery
;
Submandibular Gland Neoplasms
;
metabolism
;
pathology
;
surgery
;
Transcription Factors
;
metabolism
;
Tumor Suppressor Protein p53
;
metabolism
;
Tumor Suppressor Proteins
;
metabolism
10.Factors analysis and unstabe plaques of carotid atherosclerosis in ischemic cerebrovascular patients
Mei ZHANG ; Chuan-Qing YU ; Min XUE ; Zong WANG ; Jia-Lan DAI ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(12):-
Objective To explore the relative factors of unstable plaques of carotid atherosclerosis in ischemic eerebrovascular patients.Methods Carotid arteries of a total of 132 cases with ischemic cerebrovascular disease of carotid artery system were inspected by color Doppler ultrasound.The plaques discovered were classified according to ultrasonic appearance and their stability was judged.The relation between hypertension,diabetes,hyperlipemia, smoking and unstable plaques of carotid atheroselerosis was analyzed.Results The most common site of plaque for- mation was the bifurcate of the common carotid artery(56.99%),and the second commonest was carotid artery (23.12%).The incidence of unstable plaques in the patients with smoking,hypertension and diabetes was higher than those without them(P