Objective: To observe the clinical value of three dimensional conformal radiation therapy (3D-CRT) followed by radical surgery and discuss the best radiation technique for cervical cancer patients after radical hysterectomy. Methods: From February of 2003 to June of 2006, 155 stage I-IIIa cervical cancer patients received postoperative radiotherapy in our department after radical surgery. They were randomly divided into two groups. There were 81 patients in 3D-CRT group and 74 patients in traditional radiation group. According to FIGO staging, there were 45 cases in stage I, 77 in stage II a, 31 in stage II b, and 2 in stage III a. Pathological examination confirmed that 148 cases had squamous carcinoma and 7 cases had adenocarcinoma. The target volume included supravaginal portion, the cervical stump, paracervical tissue, common iliac, internal and external iliac, obturator, and sacral lymph nodes, and the pelvic lymphatic drainage area. For 3D-CRT group we designed four-field or two-field rotating irradiation in the left-right and the anterior-posterior direction. For traditional radiation group we designed two-field irradiation, anterior-posterior, at opposed lateral directions. The radiation dose ranged from 48-50 Gy. Stage II b patients with a cervical stump recurrence received postoperative boost irradiation by 8-10 Gy. Results: There were no significant difference in 0.5-year, 1-year, 1.5-year, and 2-year local control rate between 3D-CRT group and traditional radiation group (P > 0.05). The occurrence of early and late complications was significantly lower in 3D-CRT group than that in traditional radiation group (P < 0.05). There was significant difference in gastrointestinal reaction and urinary system reaction between the two groups (P <0.05). In postoperative radiotherapy, 3D-CRT was superior compared with traditional two-field radiation at opposed lateral directions. Conclusion: 3D-CRT is superior compared with traditional two-dimensional radiation. Four-field rotating radiation in 3D-CRT induces focused and even dose distribution and causes less side effects and complications. The side-field and cervical stump-targeted boost irradiation are apparent advantages of 3D-CRT.

Plant transgenic system secures a safe,economical and reliable supply of recombinant proteins.Plant oilbody expression system simplifies the downstream purification steps and reduces capital investment based on the nature of oleosin including high expression and easy extraction.The structures and characteristics of seed oil body and oleosin were reviewed.And the research progress and industry of the seed oil body expression system,as a new bioreactor,to produce valuable recombinant proteins were discussed.The benefits and questions of the oil body expression system were also set forth.New medicine haFGF based on the oilbody system is being developed,and its biological activity is being analyzed.As a new resource for medicine protein,oilbody expression system will be perfected and applied broadly.

Aged ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Carcinoma, Merkel Cell ; metabolism ; pathology ; Carcinoma, Small Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Lymphoma ; metabolism ; pathology ; Melanoma ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism

Objective To investigate the role of autophagy in radiation-induced death response of human nasopharyngeal carcinoma cells.Methods MTT method was used to detect cell viability of CNE-2 cells in different time after irradiation.Clonogenic survival assay was used to evaluate the effect of autophagy inhibitor (chloroquine phosphate) and autophagy inductor (rapamycin) on radiosensitivity of nasopharyngeal carcinoma cells.Cell apoptosis was assessed by flow cytometry.The expressions of LC3 and P62 were measured with Western blot.Cell ultrastructural analysis was performed under an electron microscope.Results Irradiation with 10 Gy induced a massive accumulation of autophagosomes accompanied with up-regulation of LC3-Ⅱ expression in CNE-2 cells.Compared with radiation alone,chloroquine phosphate (CDP) enhanced radiosensitivity significantly by decreasing cell viability (F =25.88,P ＜ 0.05),autophagic ratio (F =105.15,P ＜ 0.05),and LC3-Ⅱ protein level(F =231.68,P ＜0.05),while up-regulating the expression of P62 (F =117.52,P ＜ 0.05).Inhibition of autophagy increased radiation-induced apoptosis (F =143.72,P ＜ 0.05).Rapamycin (RAPA) also significantly decreased cell viability,but increased autophagic ratio and LC3-Ⅱ protein level while down-regulated the expression of P62.Induction of autophagy increased radiation-induced apoptosis(F =167.32,P ＜ 0.05).Conclusions Blockage of autophagy with CDP could enhance radiosensitivity in human nasopharyngeal carcinoma cells,suggesting that inhibition of autophagy could be used as an adjuvant treatment to nasopharyngeal carcinoma.

Objective To discover radioresistance-associated proteins by performing comparative proteomic analysis on nasopharyngeal carcinoma cell lines.Methods The total proteins were extracted from radioresistant human nasopharyngeal carcinoma cell line CNE-2R and its parental cell line CNE-2,respectively.These proteins were separated by high quality two-dimensional polyacrylamide gel electrophoresis (2-DE) and then the 2-DE profiles were screened for differentially expressed protein spots by the Image Master 5.0 software.Those spots were identified by a matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry.Results 32 significantly differentially expressed protein spots were screened in two different radiosensitivity cell lines and 11 proteins were identified by tandem mass spectrometry,among which 3 proteins were up-regulated in radioresistant human nasopharyngcal carcinoma cell line CNE-2R and the other 8 proteins were down-regulated.Conclusions The differentially expressed proteins of nasopharyegeal carcinoma cells with different radiosensitivity were mainly involved in apoptosis regulation,DNA damage and repair,cell cycle regulation,RNA transcription,cell signaling,cytoskeleton formation and radiation stress responses.

AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.

Objective To study the difference of gene expression profile between the radioresistant human nasopharyngeal carcinoma cell line CNE-2R and CNE-2,and to screen the signaling pathway associated with radioresistance of nasopharyngeal carcinoma.Methods The radioresistant nasopharyngeal carcinoma cell line CNE-2R was constructed from the original cell line CNE-2.CNE-2R and CNE-2 cells were cultured and administered with 60Co γ-ray irradiation at the dose of 400 cGy for 15 times.Human-6v 3.0 whole genome expression profile was used to screen the differentially expressed genes.Bioinformatic analysis was used to identify the pathways related to radioresistance.Results The number of the differentially expressed genes that were found in these 2 experiments was 374.The Kegg pathway and Biocarta pathway analysis of the differentially expressed genes showed the biological importance of Toll-like receptor signaling pathway and IL-1 R-mediated signal transduction pathway to the radioresistance of the CNE-2R cells and the significant differences of 13 genes in these 2 pathways,including JUN,MYD88,CCL5,CXCL10,STAT1,LY96,FOS,CCL3,IL-6,IL-8,IL-1α,IL-1B,and IRAK2(t=13.47-66.57,P<0.05).Conclusions Toll-like receptor signaling pathway and IL-1R-mediated signal transduction pathway might be related to the occurrence of radioresistance.

Objective To discover radioresistance associated molecular biomarkers and its mechanism in nasopharyngeal carcinoma by protein-protein interaction network analysis.Methods Whole genome expression microarray was applied to screen out differentially expressed genes in two cell lines CNE- 2R and CNE-2 with different radiosensitivity.Four differentially expressed genes were randomly selected for further verification by the semi-quantitative RT-PCR analysis with self-designed primers. The common differentially expressed genes from two experiments were analyzed with the SNOW online database in order to find out the central node related to the biomarkers of nasopharyngeal carcinoma radioresistance. The expression of STAT1 in CNE-2R and CNE-2 cells was measured by Western blot.Results Compared with CNE-2 cells,374 genes in CNE-2R cells were differentially expressed while 197 genes showed significant differences.Four randomly selected differentially expressed genes were verified by RT-PCR and had same change trend in consistent with the results of chip assay. Analysis with the SNOW database demonstrated that those 197 genes could form a complicated interaction network where STAT1 and JUN might be two key nodes.Indeed,the STAT1-α expression in CNE-2R was higher than that in CNE-2 (t =4.96,P ＜ 0.05).Conclusions The key nodes of STAT1 and JUN may be the molecular biomarkers leading to radioresistance in nasopharyngeal carcinoma,and STAT1-α might have close relationship with radioresistance.

Objective To investigate the relationship between genetic single-nucleotide polymorphisms of ligase Ⅳ (LIG4) and heat shock protein B1 (HSPB1) and prognosis in nasopharyngeal carcinomas.Methods DNAs were extracted by phenol-chlorofrom method from peripheral blood samples of 168 nasopharyngeal carcinoma patients.Genetic single-nucleotide polymorphisms were divided by Taqman realtime polymerase chain reaction (PCR).All statistical analyses were performed with statistical product and service solutions v13.0.A p-value of less than 0.05 was confirmed as statistical significance.Single-factor and multiple-factor logistic analyses were used to calculate odds ratio (OR) and 95％ confidence interval (CI).Long-rank test and COX were also used.Results No relationship was found between the recurrence,metastasis and overall survival of nasopharyngeal carcinoma and genetic single-nucleotide polymorphisms of LIG4 and HSPB1 with single-factor and multiple-factor logistic analyses.Conclusions LIG4 rs1805388 and HSPB1 rs2868371 had no relationship with prognosis of nasopharyngeal carcinomas.

Background Normal ultrastructure is the anatomical basis of retinal pigment epithelial(RPE) cells to perform normal physiological function.At present the precipitation method is often used to detect the ultrastructure of RPE cells with transmission electron microscopy(TEM).Objective The aim of this study was to explore a simple and feasible approach to examine the ultrastructure of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells.Methods hESCs were induced and differentiated into RPE cells by the spontaneous differentiation method,and the expressions of microphthalmia associated transcription factor MITF and paired-box gene 6 (PAX6),specific protein of RPE cells,in the cells were detected by immunofluorescence assay.hESC-RPE cells were inoculated into Transwell filter,and the ultrastructure of the cell sheet was examined under the TEM.Then the ultrastructure of the cell sheet specimens was compared with those of hESC-RPE cells from cell precipitation and RPE cell specimens of 90-day-old Long Evans rats.Results MITF and PAX6 were positively expressed in hESC-RPE cells.The normal ultrastructure were visible in the RPE cells of rats under the TEM,including apical microvilli,polarized melanin granules,cellular nucleus,basement membrane and intercellular junctions,and the ultrastructure of hESC-RPE cell sheet on Transwell was similar to the RPE cells in rats.However,only scatter melanin granules,nonpolar nucleus and scanty microvilli were observed under the TEM in the hESC-RPE cells by cell precipitation method.Conclusions Without digestion process,hESC-RPE cell sheet on Transwell can retain the normal ultrastructure of hESC-RPE cells under the TEM,with a more simple and reliable advantage.