1.Poly(ADP-ribose) polymerase 1 and its inhibitors in tumor treatment
Journal of International Oncology 2010;37(4):263-265
Poly( ADP-ribose) polymerase 1 (PARP-1) plays an essential role in DNA repair and maintenance of homeostasis and genome stability. Increased PARP-1 activity has been reported in various forms of cancer. Thus the absence of PARP-1 or the use of its inhibitors depresses DNA repair function, sensitizes cancer cells to DNA damage and enhances the therapeutic effect of radio-or chemotherapy. PARP-1 is potentially a therapeutic target of malignant tumors.
2.Reasonable Configuration Education with Assistive Devices in Sichuan
Shuwei LI ; Longwei CHEN ; Hongxia LI
Chinese Journal of Rehabilitation Theory and Practice 2012;18(3):213-214
Reasonable configuration with assistive devices is a comprehensive project that involves much discipline, interdisciplinary technology. This paper summarized World Vision training about reasonable configuration with assistive devices in Sichuan province.
3.Separation,identification and immortalization of precartilaginous stem cells from neonatal rats
Weihua HU ; Fengjin GUO ; Anmin CHEN ; Shuwei ZHANG
Chinese Journal of Tissue Engineering Research 2008;12(43):8588-8592
BACKGROUND: Precartilaginous stem cells (PSCs) have strong proliferation ability and differentiation potential,but they are instable and prone to differentiate.Importing exogenous gene could immortalize them and leave phenotype character unchanged.OBJECTIVE: To establish immortalized precartilaginous stem cells (PSCs) from neonatal SD rats in vitro for the further related research about the differentiation mechanism and clinical application of precartilaginous stem cells.DESIGN,TIME AND SETTING: Single sample observation.The study was carried out in the Department of Orthopedics.Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from October 2005 to September 2006.MATERIALS: Neonatal SD rats,irrespective of gender,24-hour old,were used for prepare PSCs.METHODS: By using LipofectamineTM 2000,a gene transfection reagent,plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigene gene (SV40Tag) was transfected into the primary cultured PSCs isolated by immuniomagnetic beads coasted with the second antibody.Colonies were isolated by puromycin selection and expanded by many passages.MAIN OUTCOME MEASURES: Biological character of PSCs; plasmid identification; biological character of transfected cells and identification; RT-PCR; growth curve.RESULTS: Immunomagnetic beads separation system obtains PSCs,which was confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive PSCs.Double restriction enzyme was cut,electrophoresis confirmed pCMV was 3 kb,SV40T was 2.3 kb.A particular anti-puromycin cell clone was acquired,which was confirmed as FGFR-3 positive PSCs.The total RNA was isolated from the positive cell clones,and a 588 bp fragment,which was specific for the SV40T antigene gene,was amplified.The transfected cells were expanded to immortalized cell strain,named as immortalized precartilaginous stem cells (IPSCs).Thepopulation doubling time of IPSCs was (22.98±2.77) hours,no significant effect of subculture,freezing and recovering had been found.CONCLUSION: Precartilaginous stem cells could be isolated from neonatal SD rats,cultured in vitro,and immortalized through the transfection of pCMVSV40T/PUR.
4.miR-146a expression and its effect on the biological characteristics in gastric carcinoma
Haibo WANG ; Yanjun LIU ; Shuwei CHEN ; Wenlong WU ; Fengchen LI
Chinese Journal of Biochemical Pharmaceutics 2017;37(1):46-50
Objective To explore the expression of miR-146a and its effect on the biological characteristics in gastric carcinoma. Methods The expressions of miR-146a in gastric cancer tissue and carcinoma adjacent tissue were detected by RT-qPCR , and human gastric cancer cell line MKN-28, SGC-7901, MKN-45 and immortalized gastric epithelial cell GES-1 were cultured,GES-1 as a reference, the expressions of miR-146a in each cell line was detected by RT-qPCR; miR-146a mimics and miR-146a control were transfected into gastric cancer cell line MKN-45, and the expression of miR-146a was detected by RT-qPCR;CCK8 was used to detect the effect of miR-146a on the proliferation of cells;flow cytometry was used to detect the effect of miR-146a on cell apoptosis. Western blot was used to detect the expressions of the apoptosis-related proteins. Results The expression of miR-146a in gastric cancer tissue was significantly lower than carcinoma adjacent tissue(P<0.01); expression of miR-146a in gastric cancer cell MKN-45 was the lowest campared to GES-1, so the gastric cancer cell MKN-45 was selected as a follow-up study. The expression of miR-146a in miR-146a mimics group was significantly higher than control group(P<0.01); CCK8results showed that the proliferation rateof miR-146a mimics group was significantly lower thanmiR-146a control on 24 hour (P<0.05),and was significantly lower than miR-146a control on 48 hour and 72 hour (P<0.01).The results of flow cytometry showed that the apoptosis rate in miR-146a mimics group was significantly higher than miR-146a control;Western blot showed that the expressions of Cleaved-caspase-3 and Bax protein in miR-146a mimics group was significantly lower than those in miR-146a control, and Bcl-2 protein expression was significantly higher than miR-146a control (P< 0.05). Conclusion The expression of miR-146a in gastric carcinoma was lower than carcinoma adjacent tissue, and the miR-146a could inhibit cell proliferation and promote cell apoptosis after transfected.
5.The construction and exploration of micro-lecture in Pathology
Shuwei YU ; Wei WEI ; Liyue HUANG ; Chen WANG
Chinese Journal of Medical Education Research 2017;16(2):159-162
[Adstract] Micro-lecture, as the main teaching form of MOOC and SPOC network teaching platform, is deeply welcomed by the majority of students. Pathology is the bridge connecting basic medicine and clin-ical medicine in medical education, which has such characteristics as many knowledge points, dispersion and obscurity, etc. Therefore, the production of micro courses is based on the knowledge points, and according to the characteristics of the knowledge points, it should also be applied reasonably according to the charac-teristics of each chapter. Adopting the teaching of self-study after class and teachers and students' discussion in class for micro courses can not only greatly improve the quality of teaching, but also develop the students' ability of independent thinking and the ability to analyze and solve problems, which transforms the teaching of pathology from pure knowledge to ability.
6.Clinical study of laparoscopic hepatectomy for hepatolithiasis
Qinqiao FAN ; Shuwei CHEN ; Wensheng CAO ; Yi YU ; Xiuhong HU
Chinese Journal of General Surgery 1997;0(06):-
Objective To explore the feaibility of laparoscopic hepatectomy(LH) for hepatolithiasis.Methods Eight patients with hepatolithiasis were treated by laparoscopic cholecystectomy(LC) and common bile duct exploration(LCBDE) and LH.Laparoscopic resection of left lateral lobe of liver was performed in 7 cases,and left hemihepatectomy in 1 case.Results Procedures were all successful with operation time of(285.00?37.42) minutes,and bleeding volume(306.25?29.73)mL.The postoperative hospital stay was(7.88?1.36) days.No complications occurred.No residual stone was found in any patient.Conclusions LH was safe and effective for hepatolithiasis,and gives better results when combined with choledoscopic stone removal.
7.A randomized controlled comparison of efficacy of 4 and 8 weeks lansoprazole treatment of gastric ulcer induced by endoscopic submucosal dissection
Haibo WANG ; Yanjun LIU ; Shuwei CHEN ; Wenlong WU ; Fengchen LI
Chinese Journal of Biochemical Pharmaceutics 2016;36(9):54-57
Objective To compare the efficacy between the treatment courses of 4 weeks and 8 weeks of lansoprazole in the treatment of endoscopic mucosal stripping ( ESD ) induced iatrogenic gastric ulcer.Methods 84 gastric adenoma or early gastric cancer patients were randomly divided into two groups after lansoprazole(30 mg/day) treatment for 4 weeks and 8 weeks.After 8 weeks of ESD, the efficacy between two groups were compared.Results 69 patients from the total 84 patients were included into the final analysis, 34 patents in 4 week-treatment group and 35 patients in 8 week-treatment group.There was no significant difference between the two groups of the ulcer stage(68% in the scar stage of 4 week-treatment group and 69% in the scar stage of 8 week-treatment group, P=0.93 ).There was no significant difference between the two groups of ulcer reduction radio (0.0081 ±0.015 in 4 week-treatment group and 0.0037 ±0.008 in 8 week-treatment group, P=0.15) 8 weeks after ESD.There was no difference in scar stage and ulcer reduction radio between 4 week-treatment group and 8 week-treatment group of patients with large ulcers ( >30 mm ) . Conclusion For ESD-induced gastric ulcer, there was no significant difference in efficacy of lansoprazole between 4 week-treatment and 8 week-treatment.It may be sufficient for proton pump inhibitors only 4 weeks’ treatment in ESD-induced gastric ulcers.
8.Mechanism of β-catenin/Akt signaling pathway in apoptosis of gastric cancer cells induced by selenite sodium
Haibo WANG ; Yanjun LIU ; Shuwei CHEN ; Wenlong WU ; Fengchen LI
Chinese Journal of Biochemical Pharmaceutics 2016;36(8):36-39
Objective To explore the mechanism of β-catenin/Akt signaling pathway in the apoptosis of gastric cancer cells induced by selenite sodium.Methods Gastric cancer cell line SGC-7901 was cultured, and 0 mol/L, 5 mol/L, 10 mol/L and 20 mol/L selenite sodium treated cells; flow cytometry was used to detect the cell apoptosis rate after 24 h;10 mol/L sodium selenite treated cells, flow cytometry was used to detect the cell cycle changes after 24 h;10 mol/L sodium selenite treated cells,protein expression ofβ-catenin,cyclin D1 and protein kinase B ( Akt) activity were detected by Western blot after 0,6,12,24 h.Results Cells apoptosis rate was significantly higher than 0 mol/L after cells was treated by 5 mol/L, 10 mol/L and 20 mol/L sodium selenite, due to cells apoptosis rate was higher by 10 mol/L sodium selenite treated than 5 mol/L and 20 mol/L, 10 mol/L sodium selenite treated cells for follow-up study;gastric cancer cells was treated by 10 mol/L sodium selenite for 24 h, compared with the control group, G0/G1 phase cells decreased, cells in S phase and G2/M phase significantly increased(P<0.05);gastric cancer cells was treated by 10 mol/L sodium selenite for 0 h, 6 h, 12 h and 24 h,protein expression of β-catenin and cyclin D1 and Survivin in 6 h, 12 h and 24 h was significantly lower than that in 0 hour (P<0.01),and with the extension of time, protein expression gradually decreased;gastric cancer cells was treated by 10 mol/L sodium selenite could significantly decrease the phosphorylation of p-Akt in 6,12,24 h, while there was no significant difference of Akt among difference time points( P <0.01).Conclusion Sodium selenite could induce apoptosis of gastric cancer cells, and the cells were arrested in S phase.The mechanism may be associated with beta-catenin/Akt signaling pathway.
9.Expression and function of BRAF-activated long non-coding RNA in colorectal cancer
Qinhao GUO ; Yan ZHAO ; Jiejing CHEN ; Jun HU ; Shuwei WANG ; Dongsheng ZHANG ; Yueming SUN
Chinese Journal of Digestive Surgery 2014;13(5):376-380
Objective To investigate the expression of BRAF-activated long non-coding RNA (BANCR) in colorectal cancer,and the influence of BANCR on the biological function of HCT116 cells.Methods Fifty-six samples of colorectal cancer specimen (including the cancer tissues and precancerous tissues) were obtained at the First Affiliated Hospital of Nanjing Medical University from March 2012 to June 2013.The expressions of BANCR in all the specimens were detected by qRT-PCR (28 cases in the BANCR-high expression group and 28 cases in the BANCR-low expression group).The relationship between the expressions of BANCR and the clinicopathological factors of colorectal cancer was analyzed.The HCT116 cells were divided into 4 groups after interfering BANCR with lentiviral-mediated shRNA-1 and shRNA-2:interference group 1 (HCT116 cells transfected with LV-shRNA-1),interference group 2 (HCT116 cells transfected with LV-shRNA-2),negative control group (HCT116 cells transfected with lentivirus vector with nonsense sequence) and blank control group (HCT116 cells cultured in RPMI 1640 medium).The proliferation,apoptosis and migration of HCT116 cells in the 4 groups were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively.The comparison between the 2 groups was analyzed by u test,and multiple groups were compared by one-way analysis of variance,repeated measurement analysis of variance and LSD-t test.Multivariate analysis was done by Logistic regression model.The difference between categorical data was compared by chi-square test.Results The relative expression of BANCR in the cancer tissues was 1.6 ± 0.4,which was significantly higher than 0.9 ± 0.7 of the precancerous tissues (u =1 020.000,P < 0.05).The result of univariate analysis showed that the high expression of BANCR was correlated with the lymph node metastasis and tumor stage (x2 =4.595,7.487,P < 0.05).The result of multivariate analysis showed that lymph node metastasis and tumor stage (stage Ⅲ-Ⅳ) were the independent risk factors influencing the high expression of BANCR(OR =4.000,5.914,95% CI:1.230-12.900,1.685-20.760,P < 0.05).The relative expressions of BANCR of the interference group 1,interference group 2,negative control group and the blank control group were 0.25 ±0.04,0.20±0.06,0.96 ±0.04,0.98 ±0.03,with significant difference among the 4 groups (F =271.610,P < 0.05).The cell proliferation rates at day 6 of the interference group 1,interference group 2 and the negative control group were 80.6% ± 7.6%,81.2% ± 5.1% and 87.9% ± 13.6%,with no significant difference among the 3 groups (F =0.559,P > 0.05).The apoptotic rates of the interference group 1,interference group 2,negative control group and the blank control group were 4.7% ± 1.7%,5.1% ± 1.1%,3.1% ± 0.6% and 2.8% ± 0.9%,with no significant difference among the 4 groups (F =2.881,P > 0.05).The numbers of transmembrane cells of the interference group 1,interference group 2,negative control group and the blank control group were 135 ± 29,107 ± 18,240 ± 24 and 245 ± 22,with significant difference among the 4 groups (F =45.194,P < 0.05).Conclusions BANCR was overexpressed in the HCT116 cells,and the BANCR overexpression was correlated with lymph node metastasis and tumor stage.BANCR can promote the migration of HCT116 cells.BANCR could be an important biomarker for the diagnosis and prognosis of colorectal cancer.
10.Mechanism of apoptosis effect induced by triptolide in PC3 cells
Hemin LI ; Shuwei YANG ; Yahui LIU ; Ying ZHOU ; Yin CHEN ; Yi ZHANG ; Yuanyuan PANG ; Qttian MU
Journal of Chinese Physician 2012;14(3):313-317
Objective To detect the treatment effect of PC3 cells with triptolide on altering the expression of genes.Methods MTT assay was used to detec the inhibition of proliferation.Apoptosis was detected by Annexin-V/PI staining.RT-PCR was used to analyze the mRNA expressions of BCL-2,BAX,PIG3,P21,FAS,CASPASE3.Results Triptolide caused a time - and dose - dependent inhibition of cell proliferation,and IC50 of 24 h and 48 h were 18.3 ng/ml and 13.5 ng/ml,respectively.Compared with the 24 h group,the low concentration of triptolide(5 ng/ml,t =1.47,P >0.05)and the high concentration of triptolide ( 160 ng/ml,t =0.91,P >0.05)had no statistical significance in 48 h group,while 10 ng/ml( t =3.26,P <0.05),20 ng/ml( t =4.21,P <0.05),40 ng/ml( t =4.09,P <0.05),80 ng/ml( t =2.91,P < 0.05 )had statistical significance.At the concentration of 18.3 ng/ml,triptolide induced PC3 cells apoptosis in a time - dependent manner.Compared with the control group,Anexin-V( + )/PI(-)was(5.42±2.21)%(t =3.52,P <0.05)in 6h,(13.51±3.37)%(t =6.53,P <0.01) 12h,(29.3 ±4.53)% ( t =8.74,P <0.01) 24 h group separately,and it had statistical significance.RTPCR showed that 18.3 ng/ml triptolide up-regulated the mRNA expression of BAX and PIG3,down-regulated P21 and BCL-2.FAS and CASPASE3 did not show obvious changes.Conclusions Triptolide inhibits the proliferation of PC3 and induces apoptosis,and the changes of BCL-2,BAX,PIG3 and P21 may play an important role in the apoptosis of PC-3 cells.