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.

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.

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

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

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.

OBJECTIVETo examine the expression of nestin and neurogenin 3 (Ngn3), the markers of pancreatic stem cells, in the human fetal pancreas.

METHODSThe human fetal pancreas tissue of 12 and 14 weeks were examined for the expression of nestin and Ngn3 using the techniques of immunofluorescence dye and RT-PCR.

RESULTSBoth nestin and Ngn3 expressed widely in 12 and 14 weeks before in human fetal pancreatic tissue. In these positive cells there was no co-expressing insulin or glucagon. There were nestin and Ngn3 co-expressing cells in ducts but not in the islets. The results of RT-PCR also indicated the expression of nestin and Ngn3.

CONCLUSIONSThere was no expression of the markers of mature endocrine cells in the nestin and Ngn3 positive cells, and they were the marks of no-differentiation cells in the human fetal pancreatic tissue.

Basic Helix-Loop-Helix Transcription Factors ; biosynthesis ; genetics ; Fluoroimmunoassay ; Humans ; In Vitro Techniques ; Intermediate Filament Proteins ; biosynthesis ; genetics ; Microscopy, Fluorescence ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Nestin ; Pancreas ; cytology ; embryology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the clinical characteristics and prognosis of second primary tumor of tongue (SPTr) after nasopharyngeal carcinoma (NPCR) treated with radiotherapy.

METHODSClinical data of 53 patients with SP7T after NPCR (group A) and 252 patients with primary tongue carcinoma (group B) were analyzed retrospectively with regard to clinical characteristics and survival rate (Kaplan-Meier); and multivariate analysis was performed using Cox proportional hazards model.

RESULTSThere was no significant difference between group A and group B ( P > 0. 05) in the presenting age, sex, tumor size, cTNM stage, tumor differentiation and the rate of distant metastasis. The overall 5-year survival rates were 41.6% in group A and 56.3% in group B (chi2 = 4.40, P = 0.0359) with a statistically significant difference between two groups. The differences of tumor location (chi2 = 61.18, P = 0.000) and rate of clinical (cN+, chi2 = 6.846, P = 0.009) or pathological lymph node metastasis (pN+, X2 = 3.993, P = 0.046) were also statistically significant between group A and group B, respectively. Multivariate analysis showed that age at presence, cTNM stage and with or without neck lymph node dissection were independent risk factors affecting survival.

CONCLUSIONSecond primary tongue carcinoma after radiotherapy for nasopharyngeal carcinoma is likely to occur on the dorsal aspect of the tongue with worse prognosis but with a lower rate of lymph node metastasis than that of primary tongue carcinoma. However, radiotherapy history is not an independent influencing factor on prognosis. Surgical resection or combined modality therapy may give a better prognosis.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Combined Modality Therapy ; statistics & numerical data ; Female ; Follow-Up Studies ; Glossectomy ; methods ; Humans ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Nasopharyngeal Neoplasms ; radiotherapy ; Neck Dissection ; Neoplasms, Radiation-Induced ; etiology ; pathology ; therapy ; Neoplasms, Second Primary ; etiology ; pathology ; therapy ; Prognosis ; Proportional Hazards Models ; Radiotherapy ; adverse effects ; Retrospective Studies ; Tongue Neoplasms ; etiology ; pathology ; therapy

OBJECTIVETo investigate the factors that influence survival of the patients with differentiated thyroid carcinoma in young people and evaluate the efficiency of unilateral lobectomy plus isthmectomy with therapeutic cervical lymph node dissection and postoperative TSH (thyroid stimulating hormone) suppressive therapy.

METHODSOne hundred and thirty-one patients under 30 years old with differentiated thyroid carcinoma treated in this hospital (14 cases no more than and 117 cases more than 16 years) from Jan. 1st, 1985 to Dec. 31st, 1997 were retrospectively reviewed. One hundred and twenty-eight patients were received only surgery and TSH suppressive therapy, and 3 patients received chemotherapy or radiotherapy because of the progressive metastasis in necks or mediastina. A multivariate analysis was performed in these patients by the Cox proportional hazard model.

RESULTSThe mean follow-time (x +/- s) of all patients were (140.86 +/- 43.76) months, with range from 20 to 229 months; Ninety-eight patients followed more than 10 years. Ten patients died of thyroid cancer. The overall 10-year survival rate was 97.18%. The 10-year survival rate for patients < or = 16 years of age and > 16 years were 75.97% and 96.57% respectively (P = 0. 0006). The 10-year survival rate for women and men were 94.91% and 93.69% respectively (P = 0.5261). The 10-year survival rates of patients with papillary thyroid carcinoma and follicular thyroid carcinoma were 93.77% and 96. 55% respectively (P = 0.8137). For patients with tumor size of < or = 1 cm, 1-4 cm and >4 cm the survival rate was 100.0%, 96.40%, and 80.67% respectively (P = 0. 0589). The 10-year survival rates of patients with or without lymph node metastasis were 88.37% and 100. 0% respectively (P = 0.0313). For patients of with or without distant metastasis, The survival rate was 96.64% or 60.00% (P = 0.0000). The 10-year survival rates with or without recurrence were 86. 67% and 95.48% respectively (P = 0. 5681). Using multivariate analysis, risk factors that independently influence survival were distant metastasis, tumor size and age.

CONCLUSIONSThe distant metastasis, tumor size and age at diagnosis were the independent factors influencing survival significantly. The status of lymph node metastasis may have certain effect on the prognosis. Unilateral lobectomy plus isthmectomy with a therapeutic cervical lymph node dissection followed by postoperative TSH suppressive therapy is a favourable model to children and young adults with DTC without distant metastasis, but to the patients with distant metastasis, their prognosis of this therapy model is disappointing.

Adenocarcinoma, Follicular ; mortality ; pathology ; surgery ; Adolescent ; Adult ; Child ; Female ; Humans ; Lymphatic Metastasis ; Male ; Papilloma ; mortality ; pathology ; surgery ; Prognosis ; Retrospective Studies ; Survival Rate ; Thyroid Neoplasms ; mortality ; pathology ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo study the diagnosis and treatment of a second primary malignant tumor induced by previous radiotherapy.

METHODSFrom March 1970 to March 1997, 108 nasopharyngeal cancer (NPC) patients who developed a second primary malignant tumor induced by radiotherapy were treated. There were squamous carcinoma 43 (39.8%), sarcoma 26 (24.1%), malignant fibrous histiocytoma 14 (13.0%), adenoid cystic carcinoma 12 (11.1%), thyroid papillary adenocarcinoma 8 (7.4%) and malignant melanoma 5 (4.6%). Fifty patients underwent operation, 32 received radiotherapy, 18 received chemotherapy and 8 received operation combined with chemotherapy.

RESULTSThe 3- and 5-year tumor-free survival rates were 64.0% and 36.0% in the operation group. They were 34.4% and 18.8% in the radiotherapy group.

CONCLUSIONSurgery, if not contra-indicated, is the first choice for the second primary malignant tumor induced by radiotherapy. Aggressive treatment for these patients is, hence, indicated clinically.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasms, Radiation-Induced ; diagnosis ; mortality ; therapy ; Neoplasms, Second Primary ; diagnosis ; mortality ; therapy ; Radiotherapy ; adverse effects ; Survival Rate

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