1.Study on protective effect of the extract of ginkgo biloba on myocardial tissue in aged rat
Yan GUO ; Miao LU ; Jin QIAN ; Yunlin CHENG
Chinese Journal of Geriatrics 2009;28(3):243-246
Objective To investigate the protective effect of the extract of ginkgo biloba on myocardial tissue in aged rat.Methods The rats aged 20 months were given the extract of ginkgo biloba (EGB) and ALT-711 respectively by garage for 16 weeks.The aged controls and adult rats were infused with the same volume saline.The superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malonyt diadehyde (MDA) in blood samples and advanced glycation end products (AGEs) in the myocardial tissue were measured .The myocardial histopathological changes under electron microscope and the mitochondrial DNA (mtDNA) deletion rate in myocardial cells were observed.Results Compared with the adult rats, the content of AGEs in myocardial cells in aged rats was significantly increased [(33.5±1.3)AU/mgHYP us.(18.1±1.2)AU/mgHYP, t= 7.18,P<0.05] and the levels of SOD and GSH-PX in blood samples were decreased [(138.4±3.8) U/mlvs.(227.7±13.8)U/ml, (1283.8±28.8) U/ml vs.(2114.1 ±135.9)U/ml, t=-19.59, -18.79;both P<0.01].The MDA level in the serum and mtDNA deletion rate in aged rats were higher than in adult rats[(6.7±0.6) mmol/ml vs.(4.1±1.0) mmol/ml, (0.18054±0.0718) % vs.( 0.0060±0.0001)%, t=7.18,6.98;both P<0.05].Compared with the aged controls, the content of AGEs in myocardial cells, the level of MDA and mtDNA deletion rate were significantly decreased in EGB and ALT-711 treatment adults (all P<0.05).The SOD and GSH-PX in blood samples were increased in EGB and ALT-711 treatment adults (all P<0.05).Conclusions Nonenzymatic glycation may play an important role in myocardial aging, which may be amplified by oxidative stress.EGB and ALT-711 may have the same anti-aging effects by inhibiting nonenzymatic glycation and oxidative stress.
2.Gene expression profiling of a radioresistant esophageal squamous cancer cell line
Lingran ZHOU ; Lu BAI ; Liang WANG ; Min CHENG ; Liting QIAN
Chinese Journal of Radiation Oncology 2017;26(2):215-221
Objective To establish a radioresistant esophageal squamous cancer cell line,and to identify the radioresistant genes and mechanisms.Methods The radioresistant KYSE410-res cell line was established by repeated exposure of cell line KYSE410 to radiation.The proliferation and apoptosis of esophageal squamous cancer cells were evaluated before and after radiation.The changes in gene expression of the esophageal squamous cancer cells after radiation were determined by gene microarray and analyzed by group t test.The genes with significant difference in expression after radiation were validated.Results The KYSE410-res cells had significantly enhanced proliferation and anti-apoptosis than the KYSE410 cells (all P<0.05).The result of gene microarray showed that compared with the KYSE410 cells,the KYSE410-res cells had the expression of 463 and 251 genes upregulated and downregulated by no less than 4 folds,respectively.Those genes with different expression levels after radiation were mainly responsible for cell proliferation,adhesion,signal transduction,angiogenesis,reactive oxygen metabolism,cell damage repair,and the MAPK/ERK signaling pathway.OAS2 and UBD were key proteins in the network.In the KYSE410-res cells,the expression of HLA-DQBI,MMP1,NCAM1,ZNF521,GPC6,SELENBP1,LCN15,and TFPI-2 genes measured by real-time PCR was consistent with that measured by gene microarray.Conclusions Abnormal activation of the MAPK/ERK signaling pathway,upregulated expression of OAS2 and UBD,downregulated expression of TFPI-2,and upregulated expression of MMPs may play a role in radioresistance of esophageal cancer cells.
3.Cancer stem cells in radiation resistance of esophageal cancer:role and molecular mechanism
Lu BAI ; Liang WANG ; Lingran ZHOU ; Min CHENG ; Liting QIAN
Chinese Journal of Radiation Oncology 2016;25(4):401-406
Objective To investigate the role of cancer stem cells in radiation resistance of esophageal cancer and its molecular mechanism, and to provide a theoretical basis for radiotherapy for esophageal cancer.Methods Esophageal cancer cell line TE1 was treated with 8 Gy of radiation. Esophageal cancer cell line with resistance to radiation, TE1-res, was established and screened.Cell counting was used to evaluate cell proliferation.Flow cytometry was used to determine the expression of CD44 (high) CD24(-) CD133(+) and apoptosis in cells.The colony formation assay was used to determine the colony-forming rate and cell survival curve.Bisulfite sequencing PCR was used to determine the methylation status of cancer suppressor genes.Comparison of the data was made by group t test or analysis of variance. Results Compared with TE1 cells, TE1-res cells had significantly enhanced proliferation, a significantly higher proportion of CD44( high) CD24(-) CD133(+) cells, and significantly enhanced resistance to apoptosis (mean value 20.84×105 vs.4.46×105/day, P=0.008;(38.0±2.9)%vs.(10.1±1.3)%, P=0.001;mean value 33.23% vs.10.50%, P=0.003 ) .After treatment with 8 Gy of radiation, TE1-res cells had significantly higher colony-forming rate and D0 value than TE1 cells ((14.3±2.6)%vs.(0.9±0.3)%, P=0.011;3.28 vs.2.19 Gy, P=0.125 ) .Moreover, the promoter methylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B was significantly enhanced in TE1-res cells than in TE1 cells ((89.7±4.9)%vs.(5.0±0.5)%, P=0.001;(92.3±4.7)%vs.(10.4±0.7)%, P=0.001;(90.7±3.7)%vs.(7.9±0.4)%, P=0.001;(83.4±5.7)%vs.(17.2±1.2)%, P=0.002;(90.2±
6.7)%vs.(4.4±1.2)%, P=0.002).Conclusions Cancer stem cells play an important role in radiation resistance of esophageal cancer. The resistance to radiation is closely associated with promoter hypermethylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B.
4.Relationship of vitamin D in children with sepsis/severe sepsis and outcomes in PICU
Bingru YIN ; Suyun QIAN ; Yibing CHENG ; Guoping LU ; Yimin ZHU
Chinese Journal of Emergency Medicine 2016;25(6):709-713
Objective To determine the vitamin D status in children with sepsis/severe sepsis in pediatric intensive care unit (PICU) in order to explor the association between vitamin D status and clinical outcomes,in turn to provide evidence for optimizing nutrition support.Methods It was a prospective,observational,multi-center study,carried out in patients with sepsis/severe sepsis from March 1,2013,to March 30,2014,in the PICUs of three tertiary-care children's hospitals.Total serum 25-hydroxy vitamin D [25 (OH) D] was measured by using an enzyme-linked immunosorbent assay at admission.The association of vitamin D status at admission with length of PICU length of stay,total hospital stay,in-hospital mortality,28-days mortality and costs were analyzed.Results A total of 194 patients includng 117 boys (60.3%)and 77 girls (39.7%) were enrolled.There were 96 patients with sepsis and 98 with severe sepsis.The mortality on discharge and 28 days were 6.7% and 24.2% respectively.The median vitamin D level was 9.79 ng/mL (5.32,18.46) at admission.Of them 77.8% (151/194) had vitamin D deficiency and 50.5% (98/194) had severe vitamin D deficiency.Patients with severe vitamin D deficiency,had higher mortality on discharge (P =0.011).Vitamin D status had no significant correlations with 28 days mortality,length of PICU stay,total hospital stay and costs.Conclusions More than three-quarters (77.8%) of children with sepsis/severe sepsis in PICUs had Vitamin D deficiency.Patients with severe vitamin D deficiency at admission had higher risk of mortality at discharge.
5.DNA Extraction from Rhizoma Coptidus and optimization of RAPD reaction system
Daxia CHEN ; Longyun LI ; Min QIAN ; Cheng LU
Chinese Traditional and Herbal Drugs 1994;0(08):-
Objective To study the genomic DNA extraction from Rhizoma Coptidus and optimization of RAPD reaction system. Methods Different methods, i.e. phenol method, CTAB method, low pH extraction medium with high salt, were used to genomic DNA extract from Rhizoma Coptidus. The DNA samples obtained by the above methods were tested by agarose gel electrophoresis and ultraviolet spectrometer. Results CTAB Method was considered to be an optimal technique. Based on the genomic DNA extracted by CTAB method, a reaction system suitable for Rhizoma Coptidus was established, that is, 25 ?L amplification reactions system containing 1?PCR buffer, 2 mmol/L Mg 2+, 100—150 ?mol/L dNTP, 20 ng primer, 40 ng template DNA, and 1 U Taq DNA polymerase. Conclusion CTAB Method and RAPD reaction system can be used to RAPD analysis in Rhizoma Coptidus.
7.Enlightenment of Medical Informatics Curriculum Reform of Oregon Health and Science University in the USA
Cheng QIAN ; Xiaobin CHU ; Jinyan HE ; Li GE ; Huizhen LU
Journal of Medical Informatics 2015;(8):8-11,24
The paper introduces application and development of medical information technologies and elaborates the overall reform of medical science undergraduate curriculum made by Oregon Health and Science University including the reform method and the curriculum provision after reform.It points out the enlightenment for medical informatics curriculum reform of China from the perspectives of intensif-ying utilization of computing devices, valuing interaction with patients, enhancing combination with clinical practices, etc.
8.rIL-2,TNF-?,IFN-? and anti-CD3/anti-glioma bispecific antibodies cooper atively enhancing T lymphocytes' cytotoxicity against human glioma cells
Meiqing LOU ; Yicheng LU ; Wenzhong WANG ; Qian SHEN ; Qiang HUANG ; Cheng ZHU ; Shimin YANG ; Liying LU ;
Academic Journal of Second Military Medical University 2000;0(08):-
Objective: : To observe whether cytokines rIL 2,TNF ?,I FN ? and anti CD3/anti glioma bispecific antibody(BsAb) can work coordinately, and to investigate how to further enhance cytotoxicity of T lymphocyte against human glioma cells by BsAb. Methods: There were 12 groups,contr ast method were used to analyze the effect of cytokines rIL 2,TNF ?,IFN ? to cytoxicity directed by BsAb by single and combined experiments. Cytotoxicity was assayed by standard 18 h 3H TdR incorporation release. Resul ts: rIL 2,TNF ?,IFN ? and BsAb could cooperatively enhance the cy totoxicity of effect cells( P
9.Assessment of left ventricular regional myocardial function after acute occlusion of the left anterior descending artery in rats model by two-dimensional strain echocardiography
Qian FU ; Mingxing XIE ; Jing WANG ; Xinfang WANG ; Qing Lü ; Xiaofang LU ; Lingyun FANG ; Long CHENG
Chinese Journal of Ultrasonography 2009;18(12):1071-1075
Objective To explore the value of two-dimensional strain echocardiography for quantitative assessing the change of regional left ventricular myocardial function in rats following acute myocardial infarction. Methods Sixty Wistar rats were randomly divided into two groups. The study group consisted of 50 rats with occlusion of LAD for 30-45 minutes and the sham-operated group consisted of 10 rats without occlusion of LAD. Echocardiography were performed before operation, which was defined as baseline, and 1 week, 4 weeks and 8 weeks after operation. Left ventricular internal diameter at diastole ( LVIDd) and systole < LVIDs), fractional shortening( FS), ejection fraction (EF) and left ventricular mass(LVM) were measured by anatomical M-model echocardiography. High frame rate two-dimensional images were recorded in the left ventricular short-axis views at the papillary muscle level. Peak systolic radial strain(PRS) and circumferential strain(PCS) of each segment were measured using 2-dimensional strain software. The rats were sacrificed and the infarcted size of each segment was measured using triphenyl tetrazolium chloride (TTC) after echocardiography was performed. Fibrosis of left ventricular myocardium was displayed using Van Gieson stain in 1 weeks after infarction. Results Based on the TTC findings,the left ventricle of the study group was divided into three regions:infarcted,peri-infarct and remote myocardial regions. Van Gieson stain showed fibrosis existed in all the three regions. Compared with baseline and sham-operated group, PRS and PCS of infarcted, peri-infarct and remote myocardial regions of the study group significantly decreased within 1 week after operation ( P <0. 01) and persisted for 8 weeks. PCS and PRS of infarcted, peri-infarct and remote myocardial regions of the study group in 4 weeks and 8 weeks after operation showed no significant difference when compared with those in 1 week after operation ( P >0. 01). Compared with baseline and sham-operated group,LVIDd,LVIDs and LVM of study group all increased significantly ( P <0. 05) in 4 weeks and 8 weeks after operation,and FS and EF reduced significantly ( P <0. 05). Two-dimensional strain obtained in interobserver and intraobserver both showed high agreement. Conclusions Two-dimensional strain echocardiography can assess regional function of myocardium with different perfusion in rats following acute myocardial infarction, and provides a sensitive and reliable method to follow up the process of left ventricular remodeling after myocardial infarction.
10.Protective mechanism of sirolimus pretreatment against liver ischemia-reperfusion injury in rats
Jianhua RAO ; Ling LU ; Feng CHENG ; Liyong PU ; Xiaofeng QIAN ; Xuehao WANG ; Feng ZHANG
Chinese Journal of Organ Transplantation 2009;30(12):749-752
Objective To investigate the protective effect of sirolimus pretreatment against liver ischemia-reperfusion(I/R)injury in rat model and the possible mechanism.Methods Forty-eight male SD rats were randomized into four groups (12/group):A:sham group with saline,B:sham group with sirolimus,C:saline-operated group,D:sirolimus-operated group.The rats were pretreated with either saline or sirolimus (2 mg·kg~(-1)·d~(-1))by oral gavage for two weeks.The rat partial liver model of I/R injury was established,and the samples were collected at the 24th h after the I/R The serum ALT and AST levels were determined,the histologic changes were observed by HE staining under the light microscopy,the frequency of CD4~+ CD25~+ T cells among mononuclear cells in liver tissue was analyzed by using flow cytometry,the expression of Foxp3 mRNA was detected in liver tissue by real-time PCR,and the serum TGF-β,IL-10 levels were measured by enzyme-linked immunosorbent assay (ELISA).Results Serum ALT and AST levels were significantly decreased and the histological damage was significantly alleviated in the sirolimus-operated group as compared with saline-operated group(P<0.05).The percentage of CD4~+ CD25~+ T cells among mononuclear cells in groups A,B,C,and D was(6.12±1.87)%,(22.36±6.75)%,(4.53±1.02)% and(13.29±3.16)% respectively in liver tissue The expression levels of the Foxp3 mRNA were significantly higher in sirolimus group than in saline group(P<0.05).The ELISA showed that sirolimus could significantly increase the levels of TGF-β and IL-H)(P<0.05).Conclusion Pretreatment of sirolimus can effectively protect against liver ischemia-reperfusion injury in rats,which may be related to induction of CD4~+ CD25~+ Foxp3~+ T regulator cells by sirolimus,and the increase of TGF-β and IL-10 secretion to inhibit the imflammatory response.