2.To induce uveal melanoma cells apoptosis by mixed structure bcl-2 antisense oligodeoxynucleotides multidrug resistance
Linjie GUO ; Zhongyao WU ; Sheng ZHANG
Chinese Journal of Ocular Fundus Diseases 1996;0(01):-
Objective To investigate the role of anti apoptosis gene bcl 2 in the survival of cultured uveal melanoma UM cells. Methods Antisense oligodeoxynucleotides (AS ODN) bcl 2 were delivered with cationic lipid to primary cultured UM cells. The inhibitory effect of AS ODN bcl 2 on proliferation of UM cells was examined by 3, 4,5 Dimethyliazol 2,5 diphenyl tetrazolium bromide (MTT) method. Using DNA ladder to determine if the UM cells had been apoptotic. Bcl 2 expression was detected by RT PCR and Westernblot technics. Results The effect of AS ODN bcl 2 in inhibiting the proliferation of cultured UM cells had opposite relation to dosage. It down regulated the mRNA and protein level of bcl 2 gene, and the sense ODN didn′t have this effect. Conclusion AS ODN bcl 2 can down regulate bcl 2 expression, inhibits UM cells proliferation and induces apoptosis.
3.Clinical outcomes of tibial plateau fractures (SchatzkerⅠ,Ⅱ) treated with arthroscopy-assisted operative management and locked by LCP
Bin CHEN ; Gang GUO ; Sheng ZHANG
Orthopedic Journal of China 2006;0(06):-
[Objective]To discuss the clinical outcomes of low-energy tibial plateau fractures treated with arthroscopy-assisted operative management and locked by LCP.[Method]From January 2006 to March 2007,15 patients with tibial plateau fractures(SchatzkerⅠ,Ⅱ)were reduced with arthroscopy-assisted and treated by LCP and bone defects were filled with homogeneous allograft bone.[Result]With follow-up visits for 12~26 months,all cases were healed.According to creiteria of Rasmussen,excellent were in 10 cases,good in 4 cases,fair in 1 case,with the rate of being excellent and good added up to 93%.[Conclusion]LCP fixation is an effective technique for the low-energy tibial plateau fractures treatment with the advantage of less invasion,steadier fixation and lower complication rate.Arthroscopy-assisted operative management can help to reduce reduction loss from intra-articular more precisely to reach a higher union rate.It is also a preferable method for low-energy tibial plateau fracutures treatment(SchatzkerⅠ、Ⅱ).
4.Mild hypothermia reduces and delays cell apoptosis in the hippocampal CA_1 region following ischemia-reperfusion injury
Fu-Sheng ZHANG ; Gang ZHANG ; Rui-Guo DONG ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(09):-
Objective To investigate the protective effects of mild hypothermia on cells in the CA_1 region of the hippocampus in gerbils following global ischemia-reperfusion injury(IRI),and to explore their mechanism. Methods IRI models were established in 75 gerbils.Any changes in TUNEL positive cells and the expression of Bax and Cytochrome C were then observed in normothermic(N) ,hypothermic(H)and sham(S) groups through immu- nohistochemistry methods.Results In the H group(as compared with the N group)apoptotic cells in the CA_1 sub- field of the hippocampus significantly decreased.The expression of Bax and Cyt C at 3 h,6 h and 1 d were de- creased,and the expression fastigium was delayed.Conclusion Mild hypothermia can moderate and delay cell ap- optosis,and its mechanism might be related with reducing and delaying the expression of Bax and Cyt C released by mitochondria.
6.The mechanism of synapse changes in the region around a cerebral infarct after mobility training
Lai-Xun TAN ; Sheng-Gang SUN ; Shuang-Guo ZHANG ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(07):-
Objective To investigate how mobility training affects the synapses and their functioning in the region around a cerebral infarct.Methods One hundred and fifty rats were randomly divided into 5 groups:a mo- tor training group,a normal saline group,an Ara-c inhibition group,a mevastatin group and a control group.Cere- bral infarcts were surgically induced in all 150 rats,and the level of either glial fibrillary acidic protein(GFAP)or synaptophysin,and the cholesterol content around the infarct were observed at 7,21 and 42 days using immunohisto- chemical staining and high performance liquid chromatography.Results At 7,21 and 42 days after the infarct model was induced,significant differences in the optical density of either GFAP or synaptophysin and in cholesterol levels were noted between the motor training group and the control group.Ara-c inhibition was also significantly high- er in the controls.The optical density of synaptophysin and the cholesterol levels were significantly lower in the me- vastatin group than in the motor training group.Conclusion Motor skill training can improve synapse redefinition in rats with a model of acute cerebral infarct.Astrocytes may play a crucial role by means of the secretion of choles- terol in the region around the infarct.
7.Mechanism of hsa-miRNA124-3p regulating the proliferation and migration of human lung cancer cell line NCI-H460
Ye WANG ; Wang XIE ; Jie ZHANG ; Jieying SHENG ; Zhongliang GUO
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(8):1051-1058
Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism. Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 (KLF4) levels. The theoretical binding site of hsa-miRNA124-3p in 3'-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method. Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue (P<0.01), and KLF4 protein was lower in the cancer tissues than in the adjacent tissue (P<0.01). The bioinformatic analysis showed there is a theoretical binding site (5'-UGCCUUAA-3') of hsa-miRNA124-3p in 3'-UTR of KLF4. Luciferase activity assay showed that hsa-miRNA124-3p could bind to the 3'-UTR region of KLF4 gene and negatively regulate the expression of protein. The proliferation of NC-H460 cells was suppressed by transfection with pshRNA-Sponge-miRNA12472 h after transfection (P<0.05 ). Compared with the control group, the proliferation activity of pshRNA-KLF4 transfection group was further enhanced (P<0.05) There was no significant difference in the proliferation of pshRNA-Sponge-miRNA124 and pshRNA-KLF4 cotransfection group and the control group (P>0.05). The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection. Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migration via upregulating KLF4 expression.
8.Effects of YinlingⅠon Cell Viability and Oxidative Injury of Lead-Exposed Vero Cell
kai, SHENG ; guo-cheng, ZHANG ; dong-liang, XU
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To study effects of yinlingⅠon cell viability and oxidative injury of lead-exposed cell.Methods After lead exposure Vero cell was treated with yinlingⅠof different concentrations.The cell viablitity was measured by methyl thiaxiolyl tetrazolium(MTT) method and the superoxide dismutase(SOD)and malondialdehyde(MDA) activity in cell suspensions were measured.Results When Vero cell lived in 125 ?mol/L lead acetate surrounding,the MDA concentration increased,but the viability of cell and the SOD content in the Vero cell suspension decreased.YinlingⅠcould increase the viability of lead-exposed cell during a certain extent;in the most non-toxicity concentration yinlingⅠcould elevate the SOD content in the Vero cell suspension,reduced the MDA concentration and resisting lead toxication in vitro.Conclusion YinlingⅠhas the protective effects on the cell viability and oxidative injury of lead-exposed cell.
9.The treatment of lumbar intervertebral infection via single-stage posterior midline incision and bilateral muscle gap ap-proach
Jian ZHANG ; Hailong GUO ; Long LI ; Weibin SHENG
Chinese Journal of Orthopaedics 2016;36(11):709-716
Objective To evaluate the effect of treatment of the intervertebral infection via single?stage posterior midline incision and bilateral muscle gap approach. Methods A retrospective of 39 cases (male 25 cases, female 14 cases) of lumbar in?tervertebral infected patients from October 2012 to December 2014 who were treated by posterior debridement, interbody fusion using allograft and posterior instrumentation through paraspinal muscle gap approach were analyzed, whose mean age was 48 years (range 11-70 years). According to the confirming diagnosis, patients underwent postoperatively anti?inflammatory or chemotherapy treatment. The disease controlling statues were evaluated based on laboratory results of ESR, CRP;Imaging examinations were tak?en to evaluate the fusion of vertebral body;Clinical effects were evaluated using the Visual Analog Scale (VAS) and the JOA score of lumbar function. Results In these 39 cases of intervertebral infection patients, 8 cases ware diagnosed as pyogenic infec?tious, 25 cases were diagnosed as tuberculosis infections, 2 cases were diagnosed as unknown infections, and brucellosis infec?tious was found in 4 cases. All patients' symptoms were significantly improved. The lower back VAS score: average 8.22±0.93 points before operation, average 2.21 ± 0.88 points one week after operation, and an average score of 0.80 ± 0.58 points by the last follow?up time. The lower extremity VAS score: average 2.32 ± 1.82 points before operation, average 1.89 ± 0.62 points one week after operation, and an average score of 0.61±0.47 points by the last follow?up time. All patients were followed up for 12-18 months (average 13 months), One patient with pyogenic infectious occurred wound infection 1 week postoperatively, and healed after a repeatedly surgery. No internal fixation loosening, fracture, or segmental collapse was observed ,and good fusion was present in all patients after 12 months. JOA lumbar function score: all patients were effective after operation, the improve?ment rate was excellent in 76.9%, good in 17.9%, and passable in 5.2%. Comparing with preoperation, the excellent and good rate was 94.8%. Conclusion The treatment of lumbar intervertebral infection via single?stage posterior midline incision and bi?lateral muscle gap approach was clinically effective, which can completely remove the lesion, and achieve rigid internal fixation.
10.Ischemia reperfusion-induced proteomic changes in rat skeletal muscle
Zhenying ZHANG ; Xiuhua LIU ; Sheng SUN ; Xiaosun GUO
Chinese Journal of Pathophysiology 2000;0(11):-
AIM:To investigate ischemia reperfusion (I/R)-induced proteomic changes in rat skeletal muscle. METHODS: Healthy male Wistar rats were randomly divided into two groups as follows (n=6): sham group and I/R group. I/R of right hind limb was induced by 4 h ischemia followed by 24 h reperfusion. The 2-DE was applied to separate the proteins extracted from skeletal muscle tissue at the end of experiment, followed by Coomassie Brillant blue R-250 staining. Computer image analysis was used to determine the differential expression of proteins between the two groups, and 7 protein spots expressed differentially were picked out and subjected to in-gel digest and MALDI-TOP for identification. RESULTS: 354?13 proteins were detected and the match rate was (78.7?1.4)%. 10 proteins displayed significant changes after I/R, of which, 6 proteins increased and 3 proteins decreased in expression. Moreover, 2 spots in I/R group were observed, only 1 spots of which in control. 5 proteins were identified after mass spectrometry. Mitochondrial aldehyde dehydrogenase (ALDH) precursor, heat shock 27 kD protein (HSP27), an unnamed protein product (increased in I/R group), ?-actin (decreased in I/R group), and nuclear transport factor 2 (NTF-2) W7a mutant were found in I/R group. CONCLUSION: I/R injury induced differential proteomic changes in rat skeletal muscle. ALDH, ?-actin and HSP27 expression, and NTF-2 mutation are involved in I/R injury.