1.The effects of arsenic trioxide on epidermal growth factor-induced proliferation and migration of retinal pigment epithelial cell
Shao-bo, ZHANG ; Zhong-lou, ZHOU ; Min, SUN ; Chun-li, CHEN ; Zong-ming, SONG
Chinese Journal of Experimental Ophthalmology 2012;30(6):520-524
Background Growth factor-induced proliferation and migration of retinal pigment epithelium (RPE) cells are the major pathological changes of proliferative vitreoretinopathy (PVR).Arsenic trioxide ( As2O3 ) is an active ingredient of Chinese traditional medicines,which has an inhibition on proliferation and migration of tumor cells.However,it is not clear whether As2O3 could inhibit growth factor-induced proliferation and migration of RPE cells. Objective This study was to explore the effects of As2O3 on epidermal growth factor (EGF)-induced proliferation and migration of ARPE-19 cells. Methods RPE cell line (ARPE-19 cells) were cultured.Different concentrations of As2O3(0,0.5,1.0,2.0,5.0,10.0,20.0 μmol/L) were added in the culture plate to treat ARPE-19 cells with or without 10 mg/L EGF in serum-free group for 24 and 48 hours,respectively.The MTT colorimetric assay was used to check the cell viability and evaluate the drug toxicity.The effects of As2O3 on EGF-induced proliferation of ARPE-19 cells were analyzed to get an effective and avirulent concentrations of As2O3.The effects of As2O3 on EGF-induced migration of ARPE-19 cells were observed by scratch-wound assay and the Boyden chamber assay.Results MTT assay showed that the A values were gradually declined with the increase of As2O3 concentrations after As2O3 treatment without EGF for 24 hours and 48 hours ( Fgroup =38.269,P =0.000 ; Ftime =0.874,P =0.358 ).Compared with the control group,no significant differences were seen in the A values of ARPE-19 cells in 0.5-5.0 μmol/L groups (all P>0.05).Meantime,As2O3 reduced the A values of ARPE-19 cell with 10 mg/L EGF in dose- and time-dependent manner ( Fgroup =152.155,P =0.000 ; Ftime =51.649,P =0.000 ).There were not significant differences in 10 mg/L EGF-induced cell growth after 0.5,1.0,2.0 μmol/L As2O3 was added for 24 and 48 hours ( Fgroup =2.215,P =0.126 ;Ftime =2.230,P =0.155).However,when 5.0-20.0 μmol/L As2O3 added,the A values of 10 mg/L EGF-induced ARPE-19 cells lowed,showing a significant difference in comparison with the control groups ( all P<0.05),with the cellular inhibiting rate 12%,32%,37% in 24 hours and 39%,44% and 53% in 48 hours.Scratch-wound assay showed that EGF-induced horizontal migration of ARPE-19 cells was slow after 0.5-2.0 μmol/L As2O3 treated,and the same results also appeared in cell lognitudinal migration by Boyden chamber assay,with the inhibitory rates 22%,33% and 46% respectively. Conclusions As2O3 is avirulent on ARPE-19 cells within definite concentration range.At ≤ 2.0 μmol/L concentrations,As2O3 dose not affect EGF-induced proliferation of ARPE-19 cells,but it suppresses EGF-induced cell migration.At ≥ 5.0 μmol/L concentrations,As2O3 plays an inhibitory role to EGF-induced proliferation of ARPE-19 cells.
2.Protective effect of epidermal growth factor on proliferation and migration of the human Müller cell from oxidative damage
Chun-li, CHEN ; Zhong-lou, ZHOU ; Dong-sheng, YAN ; Jing-wei, ZHENG ; Zong-ming, SONG
Chinese Journal of Experimental Ophthalmology 2012;30(8):715-720
Background Oxidative damage plays an important role in pathogenesis of age-related macular degeneration( AMD ),and its mechanism is the destroy of blood-retinal barrier.Müller cells is a primary component to stabilize the inner barrier of the blood-retina.Researches showed that epidermal growth factor(EGF) can promote the proliferation and migration of animal Müller cells,but less study was found in the effect of EGF on human Müller cells. Objective The present study was to investigate the effects of EGF on the proliferation and migration of human Müller cells and its molecular mechanism. Methods Human Müller cell line MIO-M1 cells were cultured and incubated,and cultured cells were identified using glial fibrillory acidic protein (GFAP),factor Ⅷ,α-smooth muscle actin( α-SMA ),keratin and S-100.Different concentrations of EGF( 0,1,10,30,100 mg/L)was added in freeserum DMEM,and the positive rate of the cells was calculated using 5-bromo-2-deoxyuridine(BrdU) method.The cells were divided into EGF group,H2 O2 group,EGF + H2 O2 group,glucose oxidase ( GO ) group,GO + EGF group,EGF + LY294002+H2O2 group according to the different intervention,and the effects of LY294002 on the proliferation of Müller cells (A590 )were detected by colorimetric assay for cellular growth and survival( MTT assay).The scratch test of Müller cells was used to assess the influence of EGF(0,1,10,30,100 mg/L)on H2 O2-induced damage of human Müller cell.Western blot was used to detect the cell proliferation under the protection of EGF on co-cultured cells using LY294002 and H2O2 and the activation of Akt signal pathways. Results The proliferative rates of the cells were 28.0%,32.9%,39.0% in 10,30,100 mg/L EGF groups respectively and obviously higher than those in 0,1 mg/L EGF groups (24.5 %,26.2 % ).Under the H2O2 culture,GO culture,respectively,the A570 value of the Müller cell in high concentrations of EGF groups was significantly increased in comparison with lower concentrations EGF groups with the statistical significance among the groups( F=23.582,P=0.000).Compared with EGF+H2O2 group,the A570value of the Müller cells was lowed in EGF+LY294002+H2O2 group.The maximum migration rate of Müller cells was found in 10 mg/L EGF group.Western blot revealed that the presence of H2O2 reinforced the expression of Akt in Müller cells,however,pretreatment with 100 mg/L EGF antagonized the harmful effect of H2O2 on Müller cells.Meanwhile,pretreatment with EGF and LY294002 reduced the expression of Akt in Müller cells. Conclusions EGF can induce the proliferation and migration of human Müller cells with the strongest effect in 10 mg/L.100 mg/L exogenous EGF has a stronger protection to the Müiller cells against H2O2-induced cell damage by activating the PI3KAkt cell survival pathway.
3.Effects of pravastatin on atherosclerotic plaque and cardiovascular events in pa tients with coronary disease
Zong-Gui WU ; Jin-Ming CHEN ; Zuo HUANG ; Jun ZHAO ; Gao-Zhong HUANG ; Jia-You ZHANG ; Wei SONG
Academic Journal of Second Military Medical University 2001;22(2):161-163
Objective: To investigate the effects of pravastatin o n atherosclerotic plaque and cardiovascular events. Methods: Fifty- seven patients with coronary artery disease (44 male and 13 female, 58.4±11.3 y ears) were randommized into pravastatin and control groups. The patients in prav astatin group were administered 10 mg of pravastatin from the night of coronary angiography day. After 7.3 months (mean) of follow-up, plasma lipid parameters and coronary angiograph were repeated. Results: (1) A favorable effect on plasma lipid parameters was found. After administration, total choles terol(TC), low density lipoprotein cholesterol (LDL-C) and triglyceride(TG) red uced by 15.0% (P<0.01), 18.0% (P<0.01) and 6.0%, respectively. High den s ity lipoprotein cholesterol(HDL-C) increased by 10.6%. However, in control grou p, TC and LDL-C showed a tendency to reduce, but no significant difference was found between those of pre- and post-administration. (2)There was no significa nt difference in luminal diameter between pre- and post-administration in both groups. (3) Cardiovascular events in pravastatin group was significantly lower than those in control (P<0.05). (4) Pravastatin had no significant effect on HR, BP and left ventricular ejection fraction in both groups. Conclusio n: Pravastatin can stabilize coronary atherosclerostic plaque and reduce the incidence of cardiovascular events by improving plasma lipid parameters.
4.Effects of transforming growth factor-beta1 gene therapy on bone rarefaction around endosseous implant.
Ying-guang CAO ; Rong WANG ; Ke SONG ; Zong-qiang XIONG ; Jian-ming DU ; Hua-jun WANG
West China Journal of Stomatology 2007;25(4):335-338
OBJECTIVETo investigate the effects of transforming growth factor-beta1 (TGF-beta1) gene therapy on bone defect and bone rarefaction around endosseous implant.
METHODSThe primary cultured bone marrow derived stroma cells (BMSCs) was transfected by plasmid pCDNA3.1(+) -TGF-beta1, and was adhered with polylactic-co-glycolic acid (PLGA) for constructing TGF-beta1 gene-modified artificial bone. The model of rats with placed titanium implants in the proximal metaphyses of the tibiae after ovariectomy was made. The TGF-beta1 gene-modified artificial bone (experimental group), BMSCs-PLGA compound artificial bone (control group) and nothing (blank control group) were placed in the bone defect around implant. The tibiae were examined by decalcified sections with immunohistochemical method and histological analysis methods at intervals of 4 and 8 weeks after implant surgery in order to detect the expression of TGF-beta1 in new bone adjacent to the implant and the healing of the bone defect around the implant.
RESULTSThe expression level of TGF-beta1 of experimental group was higher than that of control group and blank control group at the 4th week. The histological analysis indicated that the gene-modified artificial bone had stronger osetogenic potential than others.
CONCLUSIONTGF-beta1 gene-modified artificial bone promotes the repair of the bone defect around titanium implants in osteoporotic rats.
Animals ; Bone and Bones ; Cells, Cultured ; Dental Implantation, Endosseous ; Dental Implants ; Female ; Genetic Therapy ; Prostheses and Implants ; Rats ; Stromal Cells ; Titanium ; Transfection ; Transforming Growth Factor beta1
5.Comparing the influencing factors of anastomotic bleeding in rectal carcinoma resection between laparoscopic and open radical approaches.
Song-ling YAN ; Zong-bin XU ; Pan CHI ; Hui-ming LIN
Chinese Journal of Gastrointestinal Surgery 2007;10(2):157-159
OBJECTIVETo observe the occurrence of anastomotic bleeding following laparoscopic and open radical resection for rectal carcinoma, and to explore its contributing factors.
METHODSTwo hundred and sixty-three cases of rectal carcinoma undergone radical resection were divided into 2 groups, laparoscopic surgery (LS) group (n=86) and open surgery (OS) group (n=177). According to the different locations of anastomotic stoma and with or without preventive colostomy, the two groups were divided into AR sub-group and LAR/UAR sub-group, colostomy sub-group and non-colostomy sub-group. After analyzing the incidence of anastomotic bleeding in each sub-group, a logistic regression model was established to determine the relationships between anastomotic bleeding and three contributing factors including surgical approaches (LS or OS), location of stoma (AR or LAR/UAR) and preventive colostomy.
RESULTSAnastomotic bleeding occurred on 16 out of 263 patients with radical resection of rectal cancer (6.1%). The rates of anastomotic bleeding in LS group and OS group were 9.3% and 4.5%, in colostomy and non-colostomy were 8.1% and 5.5%, and in AR group and LAR/UAR group were 3.3% and 12.1% respectively, there were no significant differences between them (P>0.05). Comparing the two different surgical approaches (LS vs OS), the coefficient of regression, odd ratio and standard coefficient of regression for LS were 1.319, 3.741 and 0.342 respectively. In comparison of the locations of anastomosis (AR vs LAR/UAR), the three index for LAR/UAR were 2.460, 11.704, and 0.632 respectively. Comparing colostomy with non-colostomy, the three index for colostomy were -1.394, 0.248, and -0.327 respectively.
CONCLUSIONSAnastomotic bleeding after radical rectectomy is related to the choice of surgical approach, location of anastomosis and with or without preventive colostomy. Both LS and LAR/UAR are risk factors, and preventive colostomy is a protective factor. Regarding to the significance of three factors, location of anastomosis takes the first place, following by surgical method and with or without preventive colostomy.
Adult ; Aged ; Aged, 80 and over ; Anastomosis, Surgical ; adverse effects ; Colostomy ; adverse effects ; Female ; Humans ; Laparoscopy ; adverse effects ; Male ; Middle Aged ; Postoperative Hemorrhage ; etiology ; Rectal Neoplasms ; surgery
6.Risk factors of irritable bowel syndrome in adolescents in China.
Hui-qing ZHOU ; Ding-guo LI ; Yan-yan SONG ; Chun-hua ZONG ; Ying HU ; Xiao-xing XU ; Han-ming LU
Chinese Journal of Pediatrics 2008;46(2):136-138
OBJECTIVETo explore the risk factors for irritable bowel syndrome (IBS) among school adolescents in China.
METHODA stratified, randomized study by cluster sampling was conducted, which recruited 51,956 students from high and primary schools in Chinese cities. All students were requested to fill in a questionnaire.
RESULT(1) Factors including class (odds ratio 1.12), excessive intake of pepper (odds ratio 1.17), fried (odds ratio 1.08) and starch-based foods (odds ratio 1.06), gastrointestinal tract infection (odds ratio 2.66), abuse of analgesic (odds ratio 1.49), inheritance (odds ratio 1.83), fatigue (odds ratio 1.32) and repression (odds ratio 1.45) were significantly associated with the presence of IBS (P < 0.05). High protein food (odds ratio 0.90) was a protective factor.
CONCLUSIONDifferent food intake, gastrointestinal tract infection, abuse of analgesic, inheritance and psychological factors might be related to development of IBS in the students of the cities involved in this study.
Adolescent ; Child ; China ; epidemiology ; Humans ; Irritable Bowel Syndrome ; epidemiology ; etiology ; Risk Factors ; Sampling Studies ; Students ; Surveys and Questionnaires
7.Surgical treatment of primary malignant tumors of the trachea and main bronchus.
Zhen-zong DU ; Hua REN ; Chao-ji ZHANG ; Jian-fei SONG ; Yue-pei LIANG ; Min ZHENG ; Ming DENG
Chinese Journal of Oncology 2009;31(2):152-155
OBJECTIVETo explore the clinical experience of surgical treatment of primary malignant tumors of the trachea and main bronchus.
METHODSThe clinicopathological data of 18 patients with primary malignant tumors of the trachea and main bronchus surgically treated from February 1994 to August 2007 were reviewed retrospectively. The surgical management included sleeve tracheal resection in 8 cases, lower trachea and carina resection with carina reconstruction in 4 cases, local enucleation of the tumor in 4 cases, left or right carino-pneumonectomy and carina reconstruction in 2 cases, and resection of the tracheal or bronchial tumor and reconstruction of the airway under cardiopulmonary bypass in 6 cases.
RESULTSAmong the 18 cases, there were 7 adenoid cystic carcinomas, 9 squamous cell carcinomas, 1 lymphoepithelial-like carcinoma and 1 follicular non-Hodgkin lymphoma. All the cases recovered well except one who died of endotracheal bleeding and asphyxia at the 10(th) postoperative day.
CONCLUSIONSurgical resection is the most effective treatment for primary malignant tumors of the trachea and main bronchus. The selection of operation modes should be individualized according to patients' condition. Both complete resection and safety should be taken into consideration simultaneously.
Adult ; Aged ; Bronchial Neoplasms ; surgery ; Carcinoma, Adenoid Cystic ; surgery ; Carcinoma, Squamous Cell ; surgery ; Cardiopulmonary Bypass ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tracheal Neoplasms ; surgery ; Tracheotomy ; methods ; Young Adult
8.Morphological characteristics of spermatozoa before and after renal transplantation.
Long-Gen XU ; Shi-Fang SHI ; Xiao-Ping QI ; Xiao-Feng HUANG ; Hui-Ming XU ; Qi-Zhe SONG ; Xing-Hong WANG ; Zong-Fu SHAO ; Jun-Rong ZHANG
Asian Journal of Andrology 2005;7(1):81-85
AIMTo investigate the changes of the spermatozoa ultrastructures before and after renal transplantation in uremic patients.
METHODSThe sperm of five uremic patients before and after transplantation and four healthy volunteers were collected and examined by scanning electron microscopy.
RESULTSAbnormal spermatozoa were found in patients pre-transplantation; abnormalities included deletion of the acrosome, absence of the postacrosomal and postnuclear ring, dumbbell-like changes of the head, tail curling, and absence of the mitochondrial sheath in the mid-segment. After renal transplantation, most of the spermatozoa became normal.
CONCLUSIONThere are many abnormalities with regard to the appearance and structure of the head, acrosome, mitochondria and tail of the spermatozoa in uremic patients. The majority of the spermatozoa returned to normal after renal transplantation, but a few still presented some abnormalities possibly relating to the administration of immunosuppressants.
Acrosome ; pathology ; Adult ; Case-Control Studies ; Humans ; Kidney Failure, Chronic ; complications ; Kidney Transplantation ; Male ; Microscopy, Electron ; Renal Dialysis ; Sperm Head ; pathology ; Sperm Tail ; pathology ; Spermatozoa ; pathology ; ultrastructure
9.Inhibition of bladder cancer cell growth and angiogenesis by co-blockage of vascular endothelial growth factor and its receptor KDR.
Xiu-ling CHEN ; Lu-cheng LIU ; Zong-ge XU ; Zhe LI ; Ran-wei LI ; Rui-juan GAO ; Song WANG ; Ming ZHANG ; Hang GUO
Chinese Journal of Oncology 2008;30(8):578-582
OBJECTIVETo study the effect of co-blockage of vascular endothelial growth factor (VEGF) and its receptor (KDR) on growth of bladder carcinoma T24 cells and nude mice xenograft.
METHODST24 cell line co-transfected with VEGF siRNA and sKDR expression plasmids was developed and its proliferation was assayed by MTT and apoptosis by FCM. The nude mice model bearing bladder carcinoma xenograft was established. The tumor cell VEGF expression, stroma microvessel density (MVD) and tumor cell topoisomerase II alpha (Topo II alpha) expression were detected by immunohistochemistry. Cell apoptosis was estimated by TUNEL assay.
RESULTSMTT assay showed that cell proliferation in VEGF siRNA, sKDR and combination groups was 56.3% +/- 8.3%, 42.6% +/- 13.8% and 32.5% +/- 4.3%, respectively, significantly lower than that in the scramble control (97.3% +/- 11.6%, P < 0.0001). FCM showed there were sub-diploid apoptotic peaks before G1 phase in VEGF siRNA, sKDR and combination groups, and apoptosis ratio was 5.1% +/- 0.9%, 4.2% +/- 0.5% and 8.8% +/- 0.7%, respectively, all of which were higher than that in the scramble control (0.9% +/- 0.4%, P < 0.05), and the combination group had even more higher apoptosis than the two singlely treated groups (P < 0.01). In vivo test showed that tumor growth was inhibited in VEGF siRNA, sKDR and combination groups, and from day 16 the tumor volume in combination group was significantly smaller than that in scramble control (P < 0.05), and from day 28 the tumor almost lost the ability to further growth. Immunohistochemistry revealed VEGF expression in combination group was 54.37 +/- 5.28, significantly lower than that in the scramble control (141.66 +/- 8.59, P < 0.0001). MVD number was only 8.22 +/- 3.79, much less compared with that in the scramble control (61.76 +/- 5.28, P < 0.0001) or sKDR group (19.46 +/- 4.16, P = 0.0089). Tumor cell proliferation index in the combination group (1.5% +/- 0.7%) was significantly decreased compared with that in the scramble control (11.8% +/- 5.2%, P < 0.0001), and apoptosis index (67.2% +/- 8.5%) was much higher than that in the scramble control (8.7% +/- 2.7%, P < 0.0001), VEGF siRNA group (54.3% +/- 4.8%, P = 0.0492) or sKDR group (52.3% +/- 6.4%, P = 0.0293).
CONCLUSIONVEGF siRNA or sKDR alone can inhibit tumor cell proliferation and induce cell apoptosis, but co-blockage of VEGF and KDR by their combination shows more significant therapeutic efficacy.
Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden ; Urinary Bladder Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism ; Xenograft Model Antitumor Assays
10.Study on the injurious effect of a self designed micro-skin machine on the epithelia.
Jian-she CHEN ; Jin-song CHEN ; Xian-zhi LIU ; Zong-ren ZHANG ; Guang-yu SHEN ; Hong-jie DUAN ; Yong-yue SU ; Yue-ming LIU ; Gen-fa LV
Chinese Journal of Burns 2003;19(6):355-357
OBJECTIVETo observe the injury on micro-skin induced by a self designed micro-skin machine.
METHODSMicro-skin was produced either with the machine or by hand. Cells at the edge of micro-skin were observed by transmission electron microscope. succinic dehydrogenase activity in supernatant of cultivated cells was analyzed, and the cell proliferation of micro-skin was assessed by (3)H-TdR. Twenty patients were enrolled in the study for the observation of the wound healing time between the two groups of micro-skin after being grafted.
RESULTSTransmission electron microscope examination revealed that the cellular injury at the edge of the micro-skin in machine-made group was mild compared with that in man-made group. (3)H-TdR rate was elevated but the activity of succinic dehydrogenase in the supernatant of cultured cells decreased in supernatant of cultured cells of machine produced micro-skin. Wound healing time was shortened in machine made group. (P < 0.05).
CONCLUSIONThe cellular injury at the edge of micro-skin in the machine made group was mild when compared with that in the man-made group with cell proliferation accelerated and wound healing time shortened.
Burns ; surgery ; Cell Division ; Epithelium ; pathology ; Humans ; Microscopy, Electron ; Skin ; ultrastructure ; Skin Transplantation ; methods ; Wound Healing