ObjectiveTo explore the effect of fetal bovine serum (FBS) of different concentrations in the culture medium on osteogenic growth peptide (OGP) promoting bone marrow mesenchymal stem cells (BMSCs) proliferation and differentiation. MethodsBMSCs were separated from limb bones of 8 Sprague Dawley rats (5 weeks old) and purified by adherence method, and BMSCs at passage 3 were divided into 4 groups according to OGP concentration: OGP 1×10-10 mol/L group, OGP 1×10-9mol/L group, OGP 1×10-8 mol/L group, and control group without OGP; and 0, 2%, 5%, 8%, and 10%FBS concentration gradient was used in each group. The cell proliferation rate was detected by MTT method at 1, 3, 5, 7, 9, and 12 days after culture, and the activity of intracellular alkaline phosphatase (ALP) was determined by the method of p-nitrophenyl phosphate disodium at 9 days after culture. ResultsBMSCs showed adherent growth, rapid proliferation, long fiber vortex, and typical morphology. MTT analysis showed that cells could not sustain proliferation when FBS concentration was less than 5% in each group; when FBS concentration was above 8%, cells proliferated continually. Proliferation promoting effect of OGP 1×10-8 mol/L and 1×10-9 mol/L groups was significantly higher than that of the control group in all serum concentrations (P<0.05); when FBS concentration was lower than 10%, the proliferation promoting effect of OGP 1×10-8 mol/L group was significantly higher than that of the other 2 OGP groups (P<0.05), but when FBS concentration was 10%, OGP 1×10-8 mol/L group had no advantage of promoting proliferation. ALP test results showed that as the FBS concentration increased, ALP activity of all groups also significantly increased (P<0.05). Under the condition of 5%FBS and 8%FBS, the ALP activity of each OGP group was significantly greater than that of the control group, and it was the highest in OGP 1×10-8 mol/L group (P<0.05). Under the condition of 10%FBS, the ALP activity of each OGP group was still greater than that of the control group (P<0.05), but no significant difference was found between the OGP 1×10-8 mol/L group and OGP 1×10-9 mol/L group (P>0.05). ConclusionThe concentration of 8%FBS is the best concentration of serum for OGP promoting the proliferation and differentiation of BMSCs, and the most suitable concentration of promoting the proliferation and differentiation of BMSCs is OGP 1×10-8 mol/L.
Objective To summarize the available clinical evidence on the treatment of non-proliferative diabetic retinopathy (NPDR). Methods Based on the basic methods and principles of evidence-based medicine, we searched and evaluated the NPDR-related evidence from the Cochrane Library(Issue 3,2007), PubMed (1966 to June 2007) and CBM(1979 to June 2007) Results We finally identified 1 systematic review and 20 randomized controlled trials. Clinical evidence showed that critical glycemic control and blood pressure control were essential in the treatment of NPDR, which might delay the progression of retinopathy. The effectiveness of other therapeutic measures needed to be further investigated. Conclusion NPDR is the early stage of diabetic retinopathy (DR). Relevant systematic reviews and high-quality randomized controlled trials have confirmed the effectiveness of critical control of blood glucose and blood pressure for NPDR. The effectiveness of other therapeutic measures needs to be confirmed by systematic reviews of high quality and rigorously designed randomized, multi-center and large-scale trials.
【摘要】 目的 探讨葡萄糖转运蛋白Ⅰ型（glucose transporter 1，GLUT1）和肿瘤增殖细胞核抗原Ki-67在卵巢上皮性肿瘤组织中的表达及其临床意义。 方法 收集2000年1月-2008年6月不同卵巢上皮性肿瘤病变患者119例的组织标本，采用免疫组织化学SP二步法检测肿瘤组织中GLUT1和Ki-67的表达情况。 结果 卵巢交界性、恶性上皮性肿瘤灶性或广泛高表达GLUT1和Ki-67，其表达强度有差异。卵巢良性上皮性肿瘤不表达GLUT1和Ki-67。在卵巢癌中GLUT1及Ki-67的表达强度与病理分级、临床分期、预后有关。GLUT1表达强度与病理分型无关，Ki-67表达强度与病理分型有关。 结论 卵巢上皮性肿瘤组织中GLUT1和Ki-67的表达具有相关性，其表达强度与肿瘤的良恶性质和增殖状态有关，二者同时检测可以全面了解卵巢上皮性肿瘤的性质、卵巢癌恶性程度和生物学行为，对于判断肿瘤的性质和预后有一定价值。【Abstract】 Objective To investigate the expression and clinical significance of glucose transporter-1 (GLUT1) and tumor proliferating karyon antigen Ki-67 in epithelial ovarian tumor tissue. Methods Immunohistochemistry SP method was used to detect the expression of GLUT1 and Ki-67 protein in epithelial ovarian tumor tissues from 119 patients diagnosed in our hospital from January 2000 to June 2008. Results The expressions of GLUT1 and Ki-67 had local or abroad higher expressions in the borderline and malignant epithelial ovarian tumor, and the expressive intensity was different. In benign tumors, the expression was negative. The expressive intensity of GLUT1 and Ki-67 had correlation with the grade, stage, and prognosis in malignant tumors. The expressive intensity of GLUT1 had no correlation with the type of malignant tumors, while Ki-67 related to the pathological types. Conclusion The expressions of GLUT1 and Ki-67 have relativity. The expressive intensity of GLUT1 and Ki-67 relates to the character and proliferation of epithelial ovarian tumors. The combined detection GLUT1 and Ki-67 is helpful to know the character of epithelial ovarian tumors, the malignant degree and biologic behavior of ovarian carcinoma, which is useful in estimating the character and prognosis of epithelial ovarian tumors.
Objective To study the expression of proliferating cell nuclear antigen (PCNA) in the occurrence and development of hepatocellular carcinoma. Methods Sixty SD rats were randomly divided into control group and experimental group. 3′MeDAB was administrated into rats to establish the experimental model of hepatocarcinoma. The expressions of PCNA of different phases were detected by immunohistochemistry and the liver pathologic changes were observed by optical microscope. Results The process of canceration was divided into three stages: inflammation, proliferative fibrosis and hepatic carcinoma. The expression of PCNA firstly presented in the oval cells that located in the portal area at the stage of inflammation, and a part of PCNA were hyperexpressed in the portal area. The expression rate of PCNA in the middle phase of inflammatory stage was higher than that of any other phases but declined later. Yet, when it came to the stage of hepatic carcinoma, the rate increased again. Conclusion Under the experimental circumstance when liver cancer is caused by the carcinogenic agent, PCNA may be firstly expressed in the oval cells, and the dynamic expression of PCNA may be an indicator for the early diagnosis of hepatocarcinogenesis.
Objective To observe the effects of extracellular-signal regulated kinase (ERK) 1/2 inhibitor U0126 on hepatoma carcinoma cell proliferation and apoptosis. Methods Hepatoma SMMC-7721 cell strain was divided into blank control group and different concentrations of U0126 groups. The proliferation inhibition was measured by MTT assay. FCM was used to analyze the cell cycle distribution and apoptosis. Results U0126 obviously inhibited cell proliferation, induced cell apoptosis and G0/G1 phase cell cycle arrest. There were significant differences between control group and different concentrations of U0126 groups on cell proliferation and apoptosis （P＜0.05， P＜0.01）. Conclusion Blocking ERK1/2 pathway may be an important treatment strategy for liver cancer.
【Abstract】Objective To investigate the expression of PCNA in gastric cancer and its relationship with telomerase activity of peritoneal washings and peritoneal dissemination, and to compare the efficacy of telomerase activity and cytology of peritoneal washings for prediction of peritoneal metastasis of gastric cancer. MethodsTelomeric repeated amplification protocol (TRAP)enzymelinked immunosorbent assay (ELISA) was performed to measure the telomerase activity of peritoneal washings collected from 60 patients with gastric cancer. Exfoliate cytologic analysis of the corresponding samples was used for comparison.Expression of PCNA was measured with immunohistochemical staining.Their relationship with clinicopathologic features were evaluated. ResultsThe positive rate of telomerase activity in peritoneal washing collected from patients with gastric cancer was 41.7%,which well related to serosal invasion, histology types, depth of infiltration and peritoneal metastasis of gastric cancer. The positive rate of telomerase activity increased with the increased depth of infiltration and serosal involvement areas (P＜0.05).The positive rate of exfoliative cytology was 25.0%, which was obviously high in the group with macroscopic peritoneal metastasis (the group of P1-3). The positive rate of exfoliative cytology also increased with the increased depth of infiltration and serosal involvement areas (P＜0.05). Although the positive rate of telomerase activity in peritoneal washing collected from patients with gastric cancer was not significantly higher than that of exfoliative cytology in general, it was significantly higher than that of exfoliative cytology in the group of pT4, P1-3 and undifferentiated type.The PCNA proliferation index (PI) of positive telomerase activity group was significantly higher than that of negative. The PCNA PI was significantly higher in the group of P1-3 and serosal invasion thanthat of P0 and without serosal invasion. ConclusionTo detect telomerase activity in peritoneal washings and to detect tumor cells by cytologic method are useful to predict subclinical metastasis to the peritoneum in patients with gastric cancer,but telomerase activity is more sensitive than the other one.Telomerase activity is well related to proliferating activity of gastric cancer,which was the very important reason of peritoneal metastasis and serosal invasion.
Objective To explore the effects of bile from patients with cholecystolithiasis on the growth of human gallbladder carcinoma cells GBC-SD and the potential correlation between cholecystolithiasis and gallbladder carcinoma. Methods Cholecystolithiasis bile (CB) and normal bile (NB) specimens were used for this study. The proliferative effects of bile were measured by methabenzthiazuron (MTT) assay and cell cycle and apoptosis were analyzed by flow cytometry. Results CB can significantly promote the proliferation of GBC-SD cells, GBC-SD proliferative index increased significantly after treated with 1% CB for 48 h （P＜0.05）.The Sphase fraction of CB 〔（49.26±8.07）%〕 increased remarkably (P＜0.05) compared with that of NB 〔（25.54±6.57）%〕, and the CB percentage of G0/G1 phase 〔（40.59±9.12）%〕 decreased remarkably (P＜0.05) compared with NB 〔(60.64±13.42）〕%. Conclusion CB can promote the proliferation of human gallbladder carcinoma GBC-SD cells.
【Abstract】Objective To explore the clinical significance of β-catenin expression in pancreatic carcinoma.Methods The immunohistochemical staining was performed to detect the expression of β-catenin in the specimens of 46 patients with pancreatic carcinoma and the results were statistically analyzed.Results The abnormal expression rate on the membrane was 54.3%, the poorer the differentiation, the higher the abnormal expression rate. The levels of the cases in whom metastasis occurred were much higher than those without metastasis. The abnormal cytoplasm expression rate was 21.7%,which had not significant correlation with the clinical indexes, such as staging, tumor size, grading and metastasis. In 23 patients who accepted intervention chemotherapy before operation, the cytoplasm expression rate in those with tumor mass smaller was 0, which was evidently lower than that of those without tumor mass change (33.3%). Moreover, the abnormal membrane and cytoplasm expression rates had remarkable concordance (63.0%).Conclusion The abnormal membrane expression of β-catenin may accelerate metastasis, and the abnormal expression of β-catenin in cytoplasm may result in cell proliferation.
ObjectiveTo observe the effects of endovascular radiation (ER) on the proliferation and apoptosis of medial smooth muscle cells (SMC) and to discuss the possible mechanisms of radiation in the prevention of vascular restenosis (RS) in rabbits after carotid endarterectomy (CEA).MethodsForty rabbits undergoing CEA were randomly divided into four groups (each group=10) and given a radiation dose of 0, 10, 20 and 40 Gy 32P respectively. Rabbits were killed on the 3rd, 7th, 14th, 28th and 56th day after operation. The specimens were collected and histopathologic examinations were done.ResultsProliferation apparently occurred in the intima and media of carotid the lumen became narrow in the control group on the 14 th, 28 th and 56 th day after operation. While in the radiation groups, proliferation was apparently suppressed and the lumen was much less narrowed (P＜0.05). The apoptosis rate of SMCs and PCNA positive cells increased on the 3rd day after operation and reached the peak on the 7th day. There was statistical difference between the ER groups and control group (P＜0.01). The effects were much more evident in 20 Gy and 40 Gy groups compared with 10 Gy group (P＜0.01).ConclusionER may prevent RS by suppressing SMC proliferation and migration as well as inducing SMC apoptosis. The effects are positively correlated with radiation doses. SMC proliferation and apoptosis occur in the early period after balloon injury, while hyperplasia of intima and medial happens later.