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 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 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.

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.

OBJECTIVETo explore type 2 diabetes prevalence and its risk factors among migrants and nonmigrants aged 35 years and older in Three Gorge Dam area in Yichang City of Hubei province, China.

METHODSA sample of 9865 rural residents (including 1949 Three Gorge Dam migrants and 7916 nonmigrants) aged 35 years old and over was selected from September to December in 2007 by the method of multi-stage cluster random sampling in Yiling district, Yichang City of Hubei province. The study subjects were assessed by interview, examination, and blood samples. Information on demographics, migrant information, lifestyle, history of diabetes and hypertension was obtained by a questionnaire interview. An overnight fasting blood specimen was collected to measure serum glucose, total cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C). Height, weight, waist circumference, hip circumference and blood pressure were measured. Oral glucose tolerance test (OGTT) was carried out for those whose fasting glucose was equal to or exceeded 6.1 mmol/L. The standardized prevalence of type 2 diabetes was calculated based on national census in the year of 2000. Multiple logistic regression analysis was used to explore the potential risk factors of type 2 diabetes.

RESULTSThe crude prevalence of type 2 diabetes among nonmigrants was 3.93% (310/7885) (male: 3.90% (129/3304), female: 3.95% (181/4581)), and that of migrants was 6.55% (127/1939) (male: 6.85% (52/759), female: 6.36% (75/1180)). The crude prevalence of type 2 diabetes among migrants was higher than that of nonmigrants (χ² = 25.10, P < 0.01 (male: χ² = 12.59, P < 0.01; female: χ² = 12.78, P < 0.01)). The prevalence of type 2 diabetes among nonmigrants was 3.87% in males and 4.15% in females. The standardized prevalence of type 2 diabetes among migrants was 6.92% in males and 6.33% in females. Logistic regression analysis showed that age (taking 35 - 44 years old as reference, 45 - 54 years old: OR = 1.45, 95%CI: 1.07 - 1.95; 55 - 64 years old: OR = 2.08, 95%CI: 1.53 - 2.84; 65 years old and over: OR = 1.85, 95%CI: 1.25 - 2.75), family history of diabetes (OR = 2.83, 95%CI: 1.70 - 4.72), overweight or obesity (overweight: OR = 1.36, 95%CI: 1.05 - 1.78; obesity: OR = 2.11, 95%CI: 1.47 - 3.01), central obesity (OR = 1.84, 95%CI: 1.39 - 2.44), abnormal triglyceride (OR = 1.54, 95%CI: 1.21 - 1.97), abnormal total cholesterol (OR = 1.40, 95%CI: 1.11 - 1.77) and abnormal LDL-C (OR = 1.82, 95%CI: 1.19 - 2.79) increased the risk of type 2 diabetes, and regular physical activity (OR = 0.57, 95%CI: 0.45 - 0.72) was the protective factor of type 2 diabetes.

CONCLUSIONThe prevalence of type 2 diabetes among Three Gorge Dam migrants was higher than that of nonmigrants. Increasing age, regular physical activity, family history of diabetes, overweight or obesity, central obesity, abnormal triglyceride, abnormal total cholesterol and abnormal LDL-C were related to type 2 diabetes.

Adult ; Aged ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Transients and Migrants

Objective To evaluate the epidemiological features and control status of hypertension in rural area of The Three Gorges. Methods A cross-sectional survey was carried out in mral area of Yiling District, Yichang City, which was located north-west bank of Xiling Gorges in 2007. A standard structure questionnaire was used to collect data on the hypertension history and treatment, social-economic status and life-styles, and so on. Blood pressure was measured by trained observers using standardized mercury sphygmomanometer. Hypertension was defined as systolic blood pressure ≥ 140 mm Hg( 1 mm Hg =0. 133kPa) and/or diastolic ≥90 mm Hg, or current treatment with antihypertensive medications. Hypertension control was defined as blood pressure levels of less than 140/90 mm Hg. Results A total of 9618 residents aged 35 years and above were included ( response rate: 81.3％ ), and 19. 7％ residents were the Three Gorges migrants. The age-adjusted prevalence of hypertension was 37.3％ for all, and 21.3％, 39. 8％,58.0％ for aged 35-44, 45-59, ≥60 years, respectively. The age-adjusted rate was higher in men for all (39.1％) and aged 35-44 years (27.0％) than in women (36.3％ for all and 17.8％ for aged 35-44years, P ＜ 0. 01 ). The age-adjusted rate of awareness, treatment and control rates of hypertension were lower in men (18.5％, 9. 0％, 0.9％, respectively) than in women (23.5％, 13.6％, 2. 0％,respectively, all P ＜ 0.01 ).Among the participants with treated hypertension, the control rate of hypertension was only 17.0％. Prevalence and rate of awareness, treatment and control of hypertension were similar between migrants and non-migrants. Conclusion The prevalence of hypertension is high, but the awareness, treatment, control rates were low in rural area of Yiling District. Efforts should be made to reduce the prevalence rate and to improve the control rate of hypertension in this area.

A new aporphine alkaloid (1), together with five known analogues (2-6), has been isolated from the branch of Litsea greenmaniana by using various chromatographic techniques. Their structures were identified by spectroscopic data analysis ( MS, IR, 1D and 2D NMR) as 2,9-dihydroxy-1,10-dimethoxy-4,5-dihydro-7-oxoaporphine (1), laurotetanine (2), N-methyllaurotetanine (3), isodomesticine (4), isocorydine (5), and norisocorydine (6). Compound 1 was a new compound, and compounds 2-6 were obtained from this plant for the first time.
Alkaloids ; chemistry ; Aporphines ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Litsea ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective To express, purify and refold recombinant luteinizing hormone releasing hormone-angiogenin(LHRH-Ang) toxin using E. coli. expression system. Methods Recombinant LHRHAng expression vector was constructed by replacing of EGF fragment in plasmid pET28a/EGF-Ang with LHRH-P Ⅱ fragment amplified from plasmid pET28/MSH-PEA0. DNA sequencing would be used to verify the correction of fused LHRH-P Ⅱ -Ang gene. Then, E. coli strain BI21 (DE3) was transformed by pET28a/LHRH-Ang vector. Expression of recombinant LHRH-Ang toxin was induced by Isopropyl-β-D-Thiogalactoside( IPTG ). Refolding effects of gradient dialysis was evaluated by SDS-PAGE. Results Prokaryotic expression vector pET28a/LHRH-Ang, containing LHRH-P Ⅱ -Ang fusion gene, was constructed by PCR amplification, restriction enzyme digestion and ligation method. Sequence correction of fusion gene was confirmed by DNA sequencing After IPGT induction, recombinant LHRH-Ang protein was expressed in BL21 ( DE3 ) as inclusion body ,it took 18.43% of total protein. Inclusion body was resolved in 8 mol/L urea and purified by DEAE-Sepharose FF column, the purity was 85%. Recombinant LHRH-Ang toxin was refolded and concentrated by gradient dialysis and PEG 20000, respectively. Conclusions Recombinant LHRH-Ang protein was expressed in E. coli and refolded successfully.

OBJECTIVETo investigate the effect of Hedgehog (HH) pathway on proliferation and in vitro tumorigenicity of gastric cancer cell lines.

METHODSThe expression of SHH, PTCH, SMO, SUFU and GLI1 in seven cell lines were tested by RT-PCR. siRNA targeting GLI1 mRNA was transfected into MKN28 cells. Cell proliferation and in vitro tumorigenicity were examined by CCK8 and soft agar colony formation test.

RESULTSSHH in six gastric cancer cell lines was up-regulated. Expression of PTCH in KATOIII cell lines and expression of SUFU in MKN28 and KATOIII were reduced. GLI1 siRNA significantly inhibited the expression of GLI1 in MKN28 cell line. Growth rate and colony formation rate of MKN28 cells treated with GLI1 siRNA were significantly lower than those of control cells (all P <0.001).

CONCLUSIONSHH signaling pathway is widely activated in gastric cancer cell lines. The activation of HH signaling pathway promotes the growth of MKN28 cells.

Cell Line, Tumor ; Cell Proliferation ; Gastric Mucosa ; cytology ; Hedgehog Proteins ; metabolism ; Humans ; Oncogene Proteins ; metabolism ; RNA, Small Interfering ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Trans-Activators ; metabolism ; Zinc Finger Protein GLI1