1.Effect of ischemic preconditioning on the expression of glial fibrillary acidic protein after ischemia-reperfusion in rats
Yuman HAO ; Zuming LUO ; Dong ZHOU ; Li GAO ; Zhong ZENG ; Zhong ZHANG ; Yan LIU
International Journal of Cerebrovascular Diseases 2010;18(9):664-667
Objective To observe the effect of focal cerebral ischemic preconditioning on the expression of glial fibrillary acidic protein (GFAP) and to investigate the significance of astrocyte activation in cerebral ischemic tolerance.Methods Thirty-six healthy male SpragueDawley rats were randomly divided into reischenmic,ischemic and control groups (n = 12 in each group) after ischemic preconditioning.The former two groups received 10 minutes middle cerebral artery occlusion (MCAO) preconditioning or sham operation 3 days before the 2-hour MCAO.The rats were killed 24 hours after the second MCAO.The control group only receivedthe two sham operations with an interval of three days.The infarct volume,histopathological changes,and GFAP expression in each group were compared.Results The infarct volume after ischemic preconditioning in the reischenmic and ischemic groups was 136.85 ± 14.51 mm3and 281.37 ± 29.93 mm3 respectively.The former was significantly reduced 53.15%compared to the latter (P =0.007).At the same time,neuronal degeneration and necrosis was reduced significantly,and GFAP expression was upregulated significantly (the mean absorbance for immunohistochemical staining in both groups was 102.66 ± 8.39 and 86.28 ± 6.19respectively,P = 0.009) after ischemic preconditioning in the reischemic group.Conclusions Focal ischemic preconditioning may induce brain ischemic tolerance and promote GFAP expression.The activation of astrocytes may be one of the mechanisms of cerebral ischemic tolerance.
2.Impact of ischemic preconditioning on dynamics of homing of endothelial progenitor cells after renal ischemia reperfusion injury
Hao LIU ; Ran WU ; Ruipeng JIA ; Bing ZHONG ; Peng YU ; Yan ZHAO
Chinese Journal of Nephrology 2013;(3):199-203
Objectives To investigate the impact of ischemic preconditioning (IPC) on dynamics of homing of endothelial progenitor cells (EPCs) after renal ischemia reperfusion injury (IR).Methods Sixty male Sprague-Dawley rats were randomly divided into three groups after right-side kidney nephrectomy:for sham-operated rats,lumbotomy without vascular clamping was performed; IR rats were clamped renal blood vessels for 40 minutes while IPC rats were pre-treated with 15 min ischemia and 10 min reperfusion.At 3,12,24 h,and 3 days after reperfusion,the pool of circulating,kidneys,lungs and spleens were harvested.The extent of renal injury was assessed by biochemical and histological examination.The dynamics of homing of EPCs was observed by flow cytometry.Results The rats in IPC group exhibited significant improvements in renal function and morphology.Compared with IR group and sham group,the number of EPCs in blood was increased in the IPC group at 12 h and 24 h after reperfusion (P < 0.05).The number of EPCs in kidney was increased at all times pointin the IPC group and IR group as compared to the sham group (P < 0.05.In addition,EPCs number was increased in IPC group compared with the IR group at 12 h and 24 h [(11.36±0.66)% vs (6.37±0.69)%,(6.31±0.70)% vs (4.40±0.60)%,all P< 0.05].Compared with IR group and sham group,the number of EPCs in the lung was increased in the IPC groups at 12 h after reperfusion [(2.95±0.66)% vs (1.78±0.59)%,(1.66±0.61)%,all P < 0.05].The number of EPCs in spleen was increased in the IPC group at 72 h as compared with the IR group and sham group [(0.55±0.06)% vs (0.34±0.07)%,(0.31±0.06)%,all P < 0.05].Conclusions Endogenous EPCs may home to injured kidney after IPC.EPCs can also gather in the lungs and spleen.
3.Practical skills promotion of students by competition of acupuncture and moxibustion
Yan ZHONG ; Qianyun YANG ; Jing SHEN ; Liang PENG ; Xiangming ZHANG ; Yubing LI ; Hao WANG
International Journal of Traditional Chinese Medicine 2010;32(4):381-
Objective This article introduced the concrete methods and benefits of promoting and holding competition of acupuncture and moxibustion in TCM universities or colleges. A regular skill competition will not only stimulate students' passion for TCM study and expand teaching methods, but also serve as a effective way to enhance practical skills of students.
4.Effects of HIF-1α on human gastric cancer cell's apoptosis in CO2 pneumoperitoneum environment
Yingxue HAO ; Hua ZHONG ; Dongzhu ZENG ; Yan SHI ; Yun RAO ; Lixin ZHOU ; Peiwu YU
Chinese Journal of General Surgery 2008;23(6):460-463
Objective To study the effect and mechanism of human gastric cancer cell's apoptosis in simulated CO2 pneumoperitoneum environment by HIF-1α. Methods We used a closed box to simulate CO2 pneumoperitoneum environment, a standard surgical insufflator maintained pressure of the box at 0,5,10 and 15 mm Hg respectively. MKN-45 cell's HIF-1α mRNA and protein expression were observed before and after silencing cell's HIF-1α by real time RT-PCR and Western blot. Cell's bcl-2/bax was studied by immunohistochemistry before and after silencing HIF-1α. Cell's apoptosis ratio was measured using Annexin V-FITC/PI double labelled staining. Results In 15 mm Hg group, MKN-45 cell's HIF-1α mRNA and protein expression were significantly higher than control group (P<0.01 ), while the difference was not significant among 0,5,10 mm Hg group and control group (P>0.05). In 15 mm Hg CO2 pressure, cell's bcl-2/bax was obviously lower than in control group (P<0.05) and apoptosis ratio was more than control group(P<0.01). When HIF-1α was silenced, cell's bel-2/bax and apoptosis ratio weren't significantly different between 15 mm Hg group and control group (P>0.05 ). Conclusions MKN-45 cell's apoptosis did not experience any alterations under 0,5,10 mm Hg.15 mm Hg CO2 pneumoperitoneum environment enhanced cell's apoptosis probably by way of HIF-1α.
5.Effects of CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line
Yingxue HAO ; Hua ZHONG ; Peiwu YU ; Chao ZHANG ; Dongzhu ZENG ; Yan SHI ; Yun RAO
Chinese Journal of General Surgery 2010;25(5):389-392
Objective To investigate the effects of different CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line. Methods Human gastric cancer cell line MNK-45 were exposed to 0,10,12 and 15 mm Hg CO2 pneumoperitoneum in vitro for 4 hrs. Cell cycle was measured by flowcytometry, the expression of CDK4 ,Cyclin D1、Rb and pRb was studied by Western-blot, and the binding ability of CDK4 and Cyclin D1 was evaluated by immunoprecipitation. Results The cell proliferation index in 15 mm Hg CO2 pneumoperitoneum group dropped significantly (27.4% ± 3. 7%) vs. (36. 4% ±3. 3%) ,P <0. 05, while that in other groups did not change significantly. The protein of CDK4、Cyclin D1and binding ability of Cyclin D1 and CDK4 dropped dramatically in the 15 mm Hg CO2 pneumoperitoneum group (0.71%±0.12%),(0.93% ±0.21%),(0.54%±0.11%),(0.18% ±0.02%) vs. (1.05% ±0.16%),(1.40% ±0.24%),(0.75% ±0.14%),(0.31% ±0.02%), all P<0.05. There were no changes of Rb in protein levels, while the phosphorylated Rb dropped obviously. Conclusion There was no obvious effects of clinical CO2 pneumoperitoneum on gastric cancer cells growth and proliferation.
6.In Vitro Study on Th1/Th2 Shift Modulated by CTLA4Ig in Psoriasis
Heng YAN ; Qingyi YE ; Fei HAO ; Dongping HUANG ; Yue MAI ; Baiyu ZHONG
Chinese Journal of Dermatology 1994;0(02):-
Objective To investigate Th1/Th2 pattern of response in psoriasis and to study the effects of CTLA4Ig on Th1/Th2 shift in psoriasis. Methods The levels of IL-2, IFN-?and IL-4 were assessed by ELISA in the supernatant of cultured peripheral blood mononuclear cells (PBMCs) from 33 patients with psoriasis vulgaris and 20 healthy blood donors. Cytokine synthesis was induced by activation with phytohemagglutinin (PHA) and staphylococcal enterotoxin B (SEB) with or without CTLA4Ig. Results Levels of IL-2, IFN-?and IL-4 were markedly increased by PBMCs from psoriatic patients incubated with SEB (P
7.Expression and Gene Polymorphism of Cytotoxic T Lymphocyte-Associated Molecule4on Peripheral Blood Mononuclear Cells from Patients with Psoriasis
Fei HAO ; Qingyi YE ; Heng YAN ; Dongping HUANG ; Baiyu ZHONG ; Qingchun DIAO
Chinese Journal of Dermatology 1995;0(03):-
Objective To investigate the status of expression and gene polymorphism of cytotoxic T lymphocyte-associated molecule4(CTLA4)on peripheral blood mononuclear cells(PBMCs)from patients with psoriasis.Methods Expression of CTLA4mRNA/antigen and polymorphism of CTLA4gene were analyzed in33and133patients with psoriasis confirmed clinically and/or pathologically,respectively.Expression of CT-LA4mRNA and antigen was detected by in situ hybridization and immmunohistochemistry.CTLA4exon1and3'untranslated region were amplified by polymerase chain reaction(PCR)and the amplified products were identified by single-stand conformation polymorphism(SSCP),restriction fragment length polymorphism(RFLP)or sequencing.Results Expression of both mRNA and antigen of CTLA4was significantly weaker on PBMCs induced by staphytococcal enterotoxin B(SEB)in patients with psoriasis than that in normal con-trols,without a regular pattern.The guanine49on exon1in association with fragment106bp of3'untranslat-ed region was shown to be linked to the susceptibility of psoriasis.Conclusion Defective translation and ex-pression of CTLA4take place in patients with psoriasis,which is possibly related to the polymorphism of CT-LA4.Our results suggest that CTLA4may be one of the candidate genes which cause autoimmunity in psoria-sis.
8.Detection of TNF-related apoptosis inducing ligand gene expression by real-time fluorescent quantitative method
Yan LIANG ; Zaixing YANG ; Hao WANG ; Jie CHEN ; Xiaojing HOU ; Renqian ZHONG
Chinese Journal of Laboratory Medicine 2008;31(7):797-800
Objective To establish a real time fluorescent quantitative revers transcripatase PCR(FQ-RT-PCR) method to detect the expression level of TNF-related apoptosis inducing ligand (TRAIL) mRNA in peripheral blood mononuclear ceils (PBMC) and determine its expression level in healthy donors, HBV-caused cirrhosis patients and PBC ones. Methods Specific primers and Taqman-MGB probe were designed and β-actin was used as endogenous control. The amplified fragment was obtained by RT-PCR. The quantitative template was constructed and then the fluorescent intensity was documented on the ABI Prism7000 analyzer. The standard curve was established, according to which, the TRAIl. mRNA levels in 30 healthy individuals, 30 patients with primary biliary cirrhosis (PBC) and 25 ones with HBV-caused cirrhosis were calculated automatically by software after the values of cycle threshold (Ct) were detected continuously during amplification. Results The linear detection range of the assay for TRAIL gene was 103 - 109 copies/ ug RNA ( r=-0.997). The coefficients of variation of both intra-and inter-assay reproducibility for high concentration samples were 5.6% and 6. 3% , respectively, and those for low concentration samples were12.5% and 14. 6%. The TRAIL mRNA expression level in PBC patients was [ (3.3±2.5)×105copies/ugRNA] significantly higher than that of healthy control [ (0.5±0.2)×105 copies/ug RNA ] (t=5.994,P <0.01). TRAIl. mRNA level of HBV-caused cirrhosis patients[ (2.1±0.9)×105 copies/ug RNA] wasalso significantly elevated (t=8.536, P<0.01). However, the difference between these two diseased groups had no significance. Conclusion We have successfully set up a FQ-RT-PCR method for detecting TRAIL gene expression and found that its expression levels of peripheral blood mononuelear cells in PBC and HBV caused cirrhosis patients are elevated, which provides a new insight into mechanism study of liver injury caused by cirrhosis.
9.A case of eumycetoma on the submaxilla due to Madurella
Jie YAN ; Jun DENG ; Fei HAO ; Bai-Yu ZHONG ; Qin-Jie LI ; Cun-Jian ZHOU
Chinese Journal of Dermatology 2003;0(08):-
Objective To investigate the clinicopathological features,diagnosis,treatment,prognosis and causative agent of a case of eumycetoma on the submaxilla.Methods A case of eumycetoma diagnosed in our department was assessed for its clinical and pathological features as well as mycologic and molecular identification.Related literature was reviewed.Results The patient was primarily characterized by swelling of the submaxilla,with multiple sinuses draining many black granules.Pathologic examination revealed a pyogenic granulomatous inflammation,and a number of lotus rhizome node-like hypha were observed in tissue samples through PAS staining.Sequence analysis of multiple loci of the isolates,including ITS 1,ITS2 and D1/D2,showed that it was mostly similar to Madurella mycetomatis with a homology of 97%.Conclusion This is a case ofeumycetoma on the submaxilla induced by a novel species of Madurella.
10.Treatment of retinal detachment after laser in situ kera-tomileusis
Ling-Yan, CHEN ; Lin, LÜ ; Yong-Hao, LI ; Shi-Yi, LI ; Hui, ZHONG ; Wang, FANG
International Eye Science 2011;11(6):947-950
AIM: To study the characteristics of retinal detachment surgery after laser in situ keratomileusis (LASIK).METHODS: Eleven eyes of ten patients that experienced rhegmatogenous retinal detachment after LASIK procedure participated in the study. The characteristics of retinal detachment, management and complications after surgery were analyzed . RESULTS: Retinal detachment was characterized by the large percentage of multiple peripheral holes (73%) and giant tears (27%). All eyes underwent sclera buckling, and three of them combined with pars plana vitrectomy (PPV) and silicone oil tamponade. Silicone oil was removed after 1 month. Retina was reattached successfully at the first retinal detachment surgery in all eyes except one that succeeded at the fourth time. One eye of LASIK flap dehiscence and one eye of corneal subepithelial opacity occurred after surgery.CONCLUSION: Patients after LASIK should be carefully examined under pupillary dilation during follow-up. Sclera buckling is necessary to most retinal detachment after LASIK, and corneal protection is important in the treatment.