1.Effects of artemisunate on anoikis resistance in human breast cancer cell MCF-7
Yu FAN ; Shu ZHENG ; Gang ZHAO
Chinese Journal of Pathophysiology 1986;0(04):-
AIM: To study the effects of artemisunate on anoikis resistance in human breast cancer cells. METHODS: After breast cancer MCF-7 cells were treated with artemisunate, the growth of anchorage independence of breast cancer MCF-7 cells was examined in soft agar colony formation, and apoptosis of breast cancer were evaluated by terminal deoxynucleatidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and DNA ladder. RESULTS: Artemisunate significantly inhibited the growth of anchorage independence of breast cancer MCF-7 cells in soft agar colon formation in a dose-dependent manner. Artemisunate induced apoptosis in breast cancer cells was conformed by DNA ladder and TUNEL assay, which was in a time-and dose-dependent manner. CONCLUSION: Artemisunate inhibits the growth of anchorage independence of breast cancer MCF-7 cells, which is related to anoikis.
3.Research progress on kidney injury induced by PM2.5 exposure
Journal of Preventive Medicine 2021;33(5):473-475
Abstract
Exposure to atmospheric PM2.5 is closely related to the morbidity and mortality of kidney diseases such as chronic kidney disease, membranous nephropathy and kidney cancer. Acute and chronic PM2.5 exposure lead to the damage of glomerular filtration and kidney tissue of mice. PM2.5 induces cellular oxidative stress, inflammatory response, endoplasmic reticulum stress, renin angiotensin system and bradykinin system activation, so that causes renal blood vessel and tissue damage, decreases glomerular filtration rate and clearance capacity, and mediates the occurrence of kidney damage and diseases. This article reviews the studies into the impact of PM2.5 on kidney and its mechanism form 2016 to 2020, so as to provide the basis for the prevention and treatment of kidney injury induced by PM2.5.
4.Effect of nuclear factor-?B on the anti-apoptosis induced by focal ischemia preconditioning in Wistar rats
Yanqiu DENG ; Gang ZHAO ; Zheng ZHAO ; Jianzhi WANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To explore the effect of nuclear factor-?B (NF-?B) on the anti-apoptosis induced by brain ischemia preconditioning (IP). METHODS: Temporary middle cerebral artery occlusion for 20 min followed three days reperfusion before 6 hours middle cerebral artery occlusion (MCAO) trancranially was used as preconditioning in Wistar rats. The protective role was evaluated by analyzing the infarct volume. The status of neuronal apoptosis was observed by TUNEL. The expression of NF?B p65 protein, the assay of SOD activity and MDA concentration were analyzed by using the methods of immunohistochemistry and cytochemistry. RESULTS: Compared to the control group, 20 min ischemic preconditioning, which did not produce neuronal damage obviously, reduced the infarct volume significantly after MCAO 6 h and obviously decreased the number of neural cell apoptosis in penumbra (P
5.Designation and silencing function of the small interfering RNA of HLA-A2
Jiansheng LIU ; Gang ZHAO ; Dan PEI ; Deyu ZHENG
Chinese Journal of Tissue Engineering Research 2014;(29):4605-4610
BACKGROUND:Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA wil prolong the survival time of transplants.
OBJECTIVE:To design smal interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2.
METHODS:Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis.
RESULTS AND CONCLUSION:According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successful y silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.
6.Safety of the bone marrow derived mesenchymal stem cells after HLA A 2 gene silencing
Suyan YAO ; Gang ZHAO ; Jiansheng LIU ; Wanli MA ; Deyu ZHENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;37(4):489-492,530
Objective To explore the safety of the human bone marrow‐derived mesenchymal stem cells (hBMSCs) after silencing of human leukocyte antigen A2 expression .Methods We divided the cells into three groups:normal cultured cells of the 8th passage served as control group , and hBMSCs after HLA A2 silencing expression of the 5th and 15th passage as experimental groups 1 and 2 ,respectively .The hBMSCs were recultured by sterile methods .The growth curve ,telomerase activation ,and expressions of P27 ,cyclin D2 and cyclin‐dependent kinase 4 (CDK4) were utilized to explore the safety of the hBMSCs induced by LV‐siRNA‐HLA A2 .The BMSCs were transplanted to the subcutaneous layer of nude mice .Tissue types were detected 24 weeks after transplantation . Results The cell curves had no obvious left or right shift in all the groups . The telomerease activation in experimental groups 1 and 2 did not significantly differ from those in control group . The expressions of anti‐oncogene P27 ,cyclin D2 and CDK4 had no obvious difference between the two experimental groups and control group , either . There was only ectopic osteogenesis 24 weeks after the BMSCs (HLA A2 gene silenced ) were transplanted to the subcutaneous layer of the nude mice .Conclusion There was no obvious evidence to support that hBMSCs had undergone change in safety after the silencing of HLA A 2 expression .
7.Comparison on eye biometry of Lenstar 900, A-scan ultrasound and keratometer in patients with cataract
Dan, HU ; Gang-Ping, ZHAO ; Jian-Hong, YU ; Xiao, ZHENG
International Eye Science 2014;(8):1440-1443
AIM:To investigate the differences among Lenstar 900, A-scan ultrasound and keratometer in measurement of axial length ( AL ) , anterior chamber depth ( ACD ) and corneal curvature ( K1 , K2 , Km ) , and evaluate the consistency of the instruments, with the purpose providing references for the clinical application of Lenstar 900.
METHODS: In this study we picked up 36 patients ( 50 eyes ) underwent cataract surgery, and lens nucleus hardness were under level IV. Before the operation, AL, ACD and K1 , K2 , Km were measured by Lenstar 900, A-scan ultrasound and keratometer respectively. The differences between the results were compared by the paired t-test. The correlation of the results was analyzed by Pearson correlation analysis, and the consistency was measured by Bland-Ahamn method.
RESULTS: The mean AL and ACD values measured by Lenstar 900 and A-scan ultrasound had no significantly statistic differences (P>0. 05). The K1, K2, Km measured by Lenstar 900 and keratometer were not significantly statistical different (P>0. 05). The results measured by these three instruments had close linearity correlation ( r>0.9, P<0. 01). The consistency of the results was well in Bland-Ahamn analysis.
CONCLUSION:The preoperatively biometric result of Lenstar 900, A - scan ultrasound and keratometer in patients with cataract are all reliable, and they can be substituted by each other. However, Lenstar 900 can not only measure AL, ACD and corneal curvature at the same time, but also cornal thickness, lens thickness, white to white, pupil size, optical axis eccentricity, retinal thickness and so on. It has a number of advantages such as non-touching, convenient and efficient, and can be recommended to use widely.
8.Effects of PER2 on proliferation, apoptosis and clockgene expressions in human oral squamous cell carcinoma SCC15 cells
Yiran AO ; Qin ZHAO ; Kai YANG ; Gang ZHENG
Basic & Clinical Medicine 2017;37(8):1133-1139
Objective To find the effect of alter-expressed PER2 on proliferation,apoptosis and other clock genes expression in human oral squamous cell carcinoma SCC15 cells.Methods Short hairpin RNA interference was used to knockdown PER2 in SCC15 human oral squamous cell carcinoma cells.Flow cytometry analysis was used to testify the cell proliferation and apoptosis.Quantitative real-time PCR was used to testify the mRNA expressions of PER3,BMAL1,DEC1,DEC2,CRY2,TIM,RORα,NPAS2,PER1 and REV-ERBα.Results The proliferation was enhanced and apoptosis was decreased after PER2 knockdown in SCC15 cells (P<0.05).The mRNA expression of PER3,BMAL1,DEC1,DEC2,CRY2,TIM,RORα and NPAS2 was significantly down-regulated,and the mRNA expression of PER1 and REV-ERBα was significantly up-regulated (P<0.05).Conclusions Clock gene PER2 plays an important role in regulating other clock genes of the clock gene network in cancer cells,PER2 knockdown can enhance proliferation and recede apoptosis of cancer cell.
9.A pilot study of using spurt,repeated cryoinjury and off-ventilator to establish rat model of acute myocardial infarction
Wenping ZHAO ; Gang LIU ; Ye ZHENG ; Kunshen LIU
Journal of Third Military Medical University 1983;0(04):-
Objective To explore a new method of establishing rat model of acute myocardial infarction by off ventilator support,spurt and repeated cryoinjury.Methods Sixty Wistar rats with male and female in half were randomized into three experimental groups according to cryoinjury time with 20 rats in each group:15-second group(cryoinjury for 5 s each time,three times in total),25-second group(5 s each time,five times in total)and 40-second group(5 s each time,eight times in total).The heart was exposed through a 1.0-to 1.5-cm left lateral thoracotomy.Cryoinjury of the LV free wall(LVFW)was performed by applying for 5 s a round copper probe of 6-mm in diameter immersed in liquid nitrogen for 5 min to reach-190 ℃,then the thoracic cavity was closed immediately.When the rats' breath restored,the cryoinjury of the LVFW in situ was repeatedly to ensure a transmural injury according to the protocol.The survival rate during operation and in 28 days after operation was observed.Also infarct size was observed by heart pathological section stained with Masson's trichrome on day 28 after operation.Results Infarct size in all three groups was larger than 20% signified by transmural injury,and completely myocardiolysis and distinct infarct boundary were observed.The survival rate was 16/20 in 15-second group,5/20 in 40-second group(P0.05 vs 15-second group and 40-second group).The infarct size in 40-second group was larger than the other two groups(standard deviation:10.564 4% vs 5.192 6%,6.496 0% in 15,25-second group,respectively)and its survival rate was lowest.The procedure of 25-second group was more complicated and had a tendency of higher death rate than that in 15-second group.Conclusion Using spurt,repeated ultra low temperature cryoinjury for 15 s may be an ideal method to establish the rat model of acute myocardial infarction.
10.Effects of extract of ginkgo biloba on human tubular epithelial-mesenchymal transition induced by transforming growth factor-?_1
Xizhen XU ; Changlong ZHENG ; Ling TU ; Gang ZHAO ; Daowen WANG
Chinese Journal of Pathophysiology 2000;0(11):-
AIM:to investigate the effects of extract of ginkgo biloba (EGB) on human tubular epithelial-mesenchymal transition induced by transforming growth factor-?1.METHODS: HK2 cells were induced to epithelial-mesenchymal transition by transforming growth factor-?1 (TGF-?1, 10 ?g/L). EGB was added into the medium of HK2 cells 2 h before TGF-?1 was added. The expressions of E-cadherin, ?-smooth muscle actin (?-SMA), NADPH oxidase p67phox and superoxide dismutase (SOD) were determined by Western blotting. Malondialdehyde (MDA) in the mediums of HK2 cells was detected. RESULTS: EGB significantly attenuated the downregulation of E-cadherin, the upregulation of ?-SMA and p67phox, the downregulation of SOD and the upregulation of MDA in HK2 cells induced by TGF-?1.CONCLUSION: EGB significantly attenuates human tubular epithelial-mesenchymal transition induced by TGF-?1, and its underlying mechanism is that EGB attenuates the upregulation of p67phox and the downregulation of SOD induced by TGF-?1.