Objective To study the inhibitive effect of adenovirus mediated CD gene and 5-FC on proliferative human retinal pigment epithelial (HRPE) cells, and to search for an effective method to take precautions against proliferative vitroretinopathy (PVR).Method Different concentrations of CD and 5-FC were added respectively to the cultured third-growth-generation HRPE cells.Transferance rate was detected by positive HRPE cells marked by X-gal and LacZ. The number of HRPE cells were counted and evaluated by methylthiazol-tetrazollium (MTT) method. Results The adenovirus mediated CD gene could be transfered into HRPE cells with a dose-dependent manner. Positive HRPE cells with CD gene could transform 5-FC to 5-Fu,which could inhibit the increase of HRPE cells effectively. No obvious bystander effect on the growth of HRPE cells was detected.Conclusions The adenovirus may introduce a foreign gene into cultured HRPE cells efficiently. It could be a good method to treat and prevent PVR by medication. (Chin J Ocul Fundus Dis，2003，19：168-171)
Objective To explore the effects of intraperitoneal chemotherapy with fluorouracil (FU) on the growth and metastasis of tumor cells in carbon dioxide (CO2) pneumoperitoneum. Methods Fifty male H-22 mice of clean grade were selected and randomly assigned into 5 groups in each group with 10: simple implantation group, pneumoperitoneum group, pneumoperitoneum and NS group, pneumoperitoneum and low concentration (5.0 g/L) of FU group and pneumoperitoneum and high concentration (10.0 g/L) of FU group. All mice were executed after 11 days to observe the weight and the implantation of tumor in abdominal wall. Then the expressions of proliferating cell nuclear antigen (PCNA) and vascular endothelial growth factor (VEGF) were detected by immunohistochemistry. Results The tumor weight was significantly higher in pneumoperitoneum and high concentration of FU group compared with other groups except pneumoperitoneum and low concentration of FU group (P＜0.05, P＜0.01 ). The inhibition rate of tumor was 64.5% in pneumoperitoneum and high concentration of FU group. The diameter of portsite implantation nodus was significantly bigger in pneumoperitoneum and NS group compared with pneumoperitoneum and low concentration of FU group and pneumoperitoneum and high concentration of FU group (P＜0.01). The expressions of PCNA and VEGF of ascites and portsite implantation nodus were significantly different in every group, respectively (P＜0.05, P＜0.01). Conclusion There is inhibitive effect of intraperitoneal chemotherapy with high concentration of FU on the growth and metastasis of S-180 tumor cells in CO2 pneumoperitoneum, which may be associated with downregulation of PCNA and VEGF expressions.
Objective To assess the efficacy and safety of mytomycin C versus 5-fluorouracil for trabeculectomy. Methods We electronically searched the Cochrane Central Register of Controlled Trials (Issue 3, 2008), MEDLINE (1966 to October 2008), EMbase (1947 to October 2008), CMBdisk (1979 to October 2008).We also handsearched relevant conference proceedings. Data were extracted by two reviewers independently using an extraction form. The Cochrane Collaboration’ s RevMan 5.0 software was used for statistical analyses. Results Nine randomized controlled trials (RCTs) involving 482 participants (495eyes) were identified. The trials enrolled three types of participants (high risk of failure, moderate risk of failure, low risk of failure). As for high risk of failure, compared with mytomycin C, 5-fluorouracil appeared to increase the rate of postoperative complications (RR –5.74, 95%CI –9.91, –1.58). No significant differences were found in postoperative mean intraocular pressure(IOP) (WMD –?2.31, 95%CI –?7.34, 2.71), success rate (RR 1.13, 95%CI 0.91, 1.39) and visual acuity ≥3-line decrease (RR 1.46, 95%CI 0.43, 4.94). As for low risk of failure, there were no significant differences in success rate (RR 1.10, 95%CI 0.99, 1.22) and postoperative complications (RR 1.00, 95%CI –6.21, 8.21). Conclusion In both groups of high risk and low risk of failure, there are no significant differences in postoperative mean IOP and success rate. However, in the group of high risk of failure, compared with 5-fluorouracil, mytomycin C appears to raise the rate of postoperative complications; the rate of reducing the eyes pressure cannot be concluded based on current evidence. However, as the number of the studied cases is rather small and the period of observation is also limited, long-term follow-up of multi-central RCTs with a larger number of cases are still needed before definite conclusions can be made. Further studies are also needed to better determine the pharmacokinetics and cost-effective analyses involving the use of the two agents for glaucoma filtering surgery.
目的觀察直腸癌術中局部植入氟尿嘧啶緩釋劑（5-FU SRI）的可行性及療效。 方法92例直腸癌患者分成治療組和對照組，治療組術中在瘤床和沿淋巴引流途徑分多點植入5-FU SRI 600 mg，對照組行常規直腸癌根治術，觀察2組患者的手術情況、毒副反應、近期并發癥及遠期療效。 結果2組的手術時間、出血量以及腹膜炎、吻合口漏、腸梗阻和切口感染發生率的差異均無統計學意義（P＞0.05）；2組患者的白細胞計數、肌酐及ALT水平治療后高于治療前（P＜0.05），但2組間差異無統計學意義（P＞0.05）。治療組腹腔局部復發率及遠處轉移率均較對照組低（P＜0.05）。 結論直腸癌術中植入5-FU SRI是安全可行的，是預防術后復發轉移的有效途徑。
Objective To study the effect of 5-fluorouracil (5-FU) induced apoptosis of the rectal carcinoma cell line HR8348 in vitro and the relationship between apoptosis induced by 5-FU and the expression of bcl-2，bcl-xl，bax and p53,and to investigate the possible mechanism of apoptosis of rectal carcinoma cell line HR8348 induced by 5-FU.Methods After treatment with 5-FU for 24 h,the apoptotic index was detected by methyl green and pyronine Y staining and by terminal deoxynucleotidyl transferase (TdT)mediated dUTP nick end labeling (TUNEL).The bcl-2,bcl-xl,bax and p53 gene expression of HR8348 cells were examined by immunohistochemical method.Results After treatment with 5-FU,the apoptotic index of experiment group was significantly increased,there was significant difference as compared with the control.Exposed to 5-FU for 12 h,24 h and 36 h,the expression of bcl-2 of HR8348 cell line remained unchanged,but the expression of bcl-xl slightly diminished,while the expression of bax was remarkly increased,the expression of p53 was not detected in both experiment and control groups.Conclusion This results indicate that 5-FU may induce apoptosis of rectal carcinoma cell line HR8348 and the possible mechanism of apoptosis induction is through upregulation of bax expression and the change of bax to bcl-xl ratio.
Objective Neuron purification is essential to procedure of various nerve cell experimental research, however, at present there is few reports on the effect of various factors on neural axons during purification. To find out a simple method of neuron purification, and to investigate the influence factors of corresponding purification culture in dorsal root gangl ion (DRG) tissue culture on β3-tubul in positive axon. Methods The DRGs were obtained from the 3 days neonatal SD rat microscopically and were made into cell suspension. Then, the amount of attached DRG neurons and non neuronal cells in poly-D-lysine (PDL) group, PDL/Laminin (PDL/LN) group and collagen-I (Col I) group was observed from 10 to 100 minutes. Then, the extension and arborization of β3-tubul in positive axons were observed after 72 hours completely randomised DRG tissue culture for the research of the influences among culture substrates (PDL, PDL/LN, and Col I), FBS (0, 5%, and 10%), 5 fluorouracil (5-Fu, 0, 20, and 40 μmol/L), and cytrarabine (Ara-C, 0, 10, and 20 μmol/L). Results Adherent cells were observed instantly after inoculation by inverted phase contrast microscope and inverted fluoresence microscope; after cell suspension was removed, adherent growth of DRGn cells and non-DRGn cells were still seen. In PDL group, the amount of NSE negative cells was significantly higher than that of NSE positive cells at 10 and 30 minutes (P lt; 0.05); the amount of NSE positive cells was significantly higher than that of NSE negative cells at 80, 90 and 100 minutes (P lt; 0.05). In PDL/LN gruop, there was no significant difference (P gt; 0.05) in the amount of NSE negative cells and NSE positive cells at 10, 20, 30, 40, and 50 minutes; the amount of NSE positive cells was significantly higher (P lt; 0.05) than that of NSE negative cells at 60, 70, 80, 90, 100 minutes. In Col I group, the amount of NSE negative cells was higher than that of NSE positive cells at 10-40 minutes, but showing no significant difference (P gt; 0.05); the amount of NSE positive cells was significantly higher (P lt; 0.05) than that of NSE negative cells at 70-100 minutes. At 72 hours after DRG tissue culture, the best result of β3-tubul in positive axon extension and arborization was obtained when the substrate level was PDL/LN, and the average length of PDL/LN level was significantly larger than that of other two substrates (P lt; 0.05). The highest number of β3-tubul in positive axon distal end was obtained at 5% concentration level of FBS (P lt; 0.05), but showing no significant differences in β3-tubul in positive axon length among three levels (P gt; 0.05). Both the most of β3-tubul in positive axon distal ends and the longest β3-tubul in positive axon average length were obtained at 0 μmol/L concentration level of 5-Fu, showing significant differences between 0 μmol/L level and 20, 40 μmol/L levels (P lt; 0.05). A similar result of β3-tubul in positive axon distal end was got at the 0 μmol/L level and 10 μmol/L level of Ara-C, which was significantly higher than that of 20 μmol/L level (Plt; 0.05). Conclusion? A purified DRG neuron suspension for neuron culture could be obtained via PDL differential attachment for 30 minutes. When DRG neuron culture, neuron special medium, PDL/LN substrate and 10 μmol/L Ara-C are recommended in β3-tubul in positive axon research.
ObjectiveTo investigate the effect of expressions of nucleoside transporters subtype (hENT1 and hENT2) on 5-fluorouracil(5-FU) cytotoxicity in breast cancer cell lines(MDA-MB-231, MDA-MB-468, SK-BR-3, MCF-7). MethodsFour breast cancer cell lines were chosen to detect the mRNA expressions of hENT1 and hENT2 by RT-PCR. Cells were incubated in the medium with a serial concentrations of 5-FU from 1.28×104 ng/L to 2.00×108 ng/L for 48 h. Then the cell proliferation in each cell line was measured by MTT assay and the IC50 was evaluated. Results①The mRNA expressions of hENT1 and hENT2 in the MDA-MB-231, MDA-MB-468, or SK-BR-3 cells were significantly higher than thoes in the MCF-7 cells(P < 0.05). The mRNA expression of hENT2 was detected in the MDA-MB-231, MDA-MB-468, or SK-BR-3 cells, not detected in the MCF-7 cells. 2MTT showed that IC50 of 5-FU in the MDAMB-231, MDA-MB-468, or SK-BR-3 cells was significantly lower than that in the MCF-7 cells(P < 0.05). There was no statistical difference of IC50 among the three lines(MDA-MB-231, MDA-MB-468, and SK-BR-3)(P > 0.05).③The three lines(MDA-MB-231, MDA-MB-468, and SK-BR-3) with lower IC50 of 5-FU highly expressed hENTs, and MCF-7 cell with the higher IC50 of 5-FU expressed less hENTs. ConclusionsThe expressions of hENTs in breast cancer cell lines can significantly influence 5-FU cytotoxic effect. It is implicated that the hENTs expressions might be the clue to the choice of nucleoside anticancer drugs in clinic.
ObjectiveTo investigate the effects of dipyridamole (DP), one of the human equilibrative nucleoside transporters (hENTs) blockers, on 5-fluorouracil (5-FU) induced cell apoptosis and cell cycle changes of the pancreatic cancer Panc-1 cell line. MethodsPancreatic cancer cell line Panc-1 was divided into hENTs blocked group and hENTs unblocked group. The hENTs blocked group was further divided into two subgroups according to the DP concentration, 5 μmol/L DP group and 10 μmol/L DP group. Each group was incubated in culture medium with or without 1.5×106 ng/L 5-FU for 24 h. Cell apoptosis and cell cycle were detected by flow cytometry. Results①The apoptosis results of each group： Incubated in culture medium with 1.5×106 ng/L 5-FU for 24 h, the apoptosis rates of 5 μmol/L DP group and 10 μmol/L DP group were higer than those of hENTs unblocked group （Plt;0.05）, and which of 10 μmol/L DP group was higer than that of 5 μmol/L DP group (Plt;0.05). Incubated in culture medium without 5-FU for 24 h, there were no significant differences of the apoptosis rates among three groups (Pgt;0.05). ②The cell cycle results of each group: Incubated in culture medium with 1.5×106 ng/L 5-FU for 24 h, the percentages of S phase cells in the 5 μmol/L DP group and 10 μmol/L DP group were less than those of hENTs unblocked group (Plt;0.05). The percentage of S phase cell of 5 μmol/L DP group reduced to 87.09% and that of 10 μmol/L DP group reduced to 74.06% as compared with hENTs unblocked group. 5-FU had little influence on G2 phase (Pgt;0.05) except for the percentage of G2 phase cells in the 5 μmol/L DP group increased significantly (Plt;0.05) as compared with the hENTs unblocked group. Incubated in culture medium without 5-FU for 24 h, there were no significant differences of the cell cycles among three groups (Pgt;0.05). ConclusionsIn pancreatic cancer Panc-1 cell, DP could enhance the cytotoxicity of 5-FU by blocking hENTs. The enhanced cytotoxicity is related to elevation of 5-FU concentration in cells, and unrelated to DP itself.