To compare the activity of RGD-TRAIL in different expression systems, RGD-TRAIL in both Escherichia coli (E.coli) and Pichia pastoris was constructed and expressed. In vitro activity of RGD-TRAIL from Pichia pastoris expression system was also analyzed. Genetic engineering techniques were used to construct recombinant plasmid pET30-rgd-trail and pHBM-rgd-trail. The recombinant protein RGD-TRAIL was purified with Ni ion affinity chromatography after induction. MTT assay, ELISA, scratch wound healing, transwell migration assay and Hoechst 33342 staining were performed to detect the effects of RGD-TRAIL on proliferation, binding activity, migration and apoptosis. The expression of apoptosis-associated proteins was detected by Western blotting. Recombinant protein RGD-TRAIL was successfully expressed in a form of inclusion body in E.coli, while expressed secretorily in Pichia pastoris. It possessed more potent cytotoxicity than RGD-TRAIL in E.coli by MTT assay. The RGD-TRAIL expressed by Pichia pastoris showed powerful binding affinity with cancer cells expressing α(v), DR4, DR5 and highly potent cytotoxicity through inducing apoptosis of cancer cells. Nuclear fragmentation was examined by Hoechst 33342 staining. Cleaved PARP and caspase-3 were also detected after incubation with RGD-TRAIL. Additionally, RGD-TRAIL inhibited migration significantly in A549 and HT1080 cells. The results demonstrate that Pichia pastoris expression system is more suitable for the recombinant protein RGD-TRAIL. Its binding affinity and antitumor activity might make RGD-TRAIL a promising candidate for cancer therapy.

This study is to investigate the effect and its possible mechanisms of lidamycin (LDM) combined with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in human non-small cell lung cancer (NSCLC) cells. MTT assay was used to determine the growth inhibition of the two ingredients on H460 cells. Apoptosis was examined by Annexin V-FITC/PI staining, flow cytometry assay and DNA-specific dye Hoechst 33342 staining. The level of TRAIL receptor and apoptosis-associated protein expression was detected by Western blotting analysis. The results showed that the IC50 value of LDM and TRAIL for H460 cells was 4.603 x 10(-10) mol x L(-1) and 915.3 ng x mL(-1) respectively, but the IC50 value of LDM was 3.064 x 10(-11) mol x L(-1) and 1.611 x 10(-11) mol x L(-1) when different concentrations of LDM was combined with 50 and 100 ng x mL(-1) TRAIL respectively. And the CDI value was less than 1. The apoptosis ratios also increased in the combination group relative to the single-agent treatment and the untreated control. Furthermore, the induction of the cleavage of PARP and the activation of Caspase-3 and Caspase-8 by the combination were more effective than LDM or TRAIL alone. At last, the level of death receptor 5 (DR5) expressions increased in a dose-dependent manner and time-related pattern. The data indicate that LDM inhibits the growth of H460 cells in vitro. DR5 induction contributes to enhancement of TRAIL-induced apoptosis by LDM in human non-small lung cancer cells.

Tea polyphenols (TPs), major biological active constituents of green tea, exert moderate and selective anticancer effects. Molecular mechanisms of TPs in cancer prevention and treatment involve multiple potential molecular targets. TPs inhibit growth factor receptor-mediated signal transduction pathway, decrease the activities of mitogen activated protein kinases and activator protein transcription factor-1, block nuclear factor-kappaB signaling pathway, reduce proteasome activity, lower overexpression of COX-2, subside dihydrofolate reductase and telomerase, and inhibit DNA methylation and matrix metalloproteinases. Furthermore, TPs enhance the inhibitory effect on the growth of cancers by traditional anticancer drugs or targeted antitumor drugs in vitro and in vivo and reverse multidrug resistances of cancer cells to vincristine, doxorubicin, and 5-fluorouracil. Besides, TPs reduce the nephrotoxicity induced by cisplatin, ameliorate irinotecan-induced side effects in the small intestine of mice, and decrease bleomycin-caused DNA damage in human leukocytes. TPs also increase antitumor activity of vaccine through immunological modulation. TPs play roles of the augmentation of antitumor effects, the reversal of multidrug resistance, and the reduction of side effects of chemotherapeutic drugs. TPs could be used as biochemical modulators in cancer therapy.

Cancer cachexia is a state of highly wasting in the terminal stages of cancer, which is characterized by the depletion of adipose tissue, muscle wasting and body weight loss.Loss of fat depots is more prominent than muscle wasting and often precedes.The enhancement of lipolysis, free fatty acids oxidation and white adipose tissue browning, the reduction of lipogenesis, lipid deposition and adipogenesisare important factors causing fat loss.Tumor necrosis factor α, interleukin-6 and zinc α2-glycoprotein are mediators involved in the process of depletion of adipose tissue, thus the drugs targetingthese mediators show a good promise for the treatment of cancer cachexia.The attention paid to the mechanism of depletion of adipose tissue contributes to the development of new therapeutic methods and drugs.

Objective To investigate the influence of human menopausal gonadotropin(HMG)administration at different phase of follicular development upon the outcome of in vitro fertilization-embryo transfer(IVF-ET)in the long-program.Methods A retrospective analysis was performed in 145 patients underwent the long program IVF-ET,who were normal in ovarian reservation but with low levels of serum leteinizing hormone(LH)(＜ 1 U/L)after the pituitary down-regulation.According to the time point of HMG administration,the patients were classified into three groups:early follicular phase(group 1,43 patients),midfollicular phase(group 2,46 patients)and late follicular phase(group 3,56 patients).The outcomes of these three groups were compared.Results Between the three groups,there was no difference in the down-regulation time,days receiving gonadotropin(Gn),the number of oocytes retrieved,day of estradiol(E2)on the day receiving chorionic gonadotrophin(hCG)injection,start date and interim LH,fertilization rate and cleavage rate (all P ＞ 0.05).In group 3,the total Gn dosage([2225 ± 292]U)was lower than that of group 1([2624 ± 422]U)(P ＜ 0.05)and group 2([2472 ± 417]U)(P ＜ 0.05).In group 1,the LH level on the day receiving hCG[(0.46 ± 0.37)U/L]was lower than that in group 2[(0.72 ± 0.58)U/L](P＜0.05).The rate of usable embryos in group 3[62.5％(288/461)]was higher than that of group 1[55.0％(170/309)]and group 252.8％(208/394)](P =0.011).Though the high qualified embryo rate,clinical pregnancy rate and implantation rate in group 3 were higher than that in goup 1 and group 2,and the abortion rate in group 1 was higher than that of group 2 and group 3,the difference was not significant(P ＞ 0.05).Conclusion For the patients with over-suppressed LH in the long-program pituitary down-regulation but with normal ovarian reservation,additional HMG during late follicular phase is helpful to improve the high qualified emryo rate,excellent rates of embryos,embryos availability,implantation rate and clinical pregnancy rate,and lower the abortion rate.

Objective To investigate whether cystatin C-based prediction equations for GFR estimation are superior to SCr-based prediction equations.Methods One hundred and ninety-eight consecutive patients (85 males,113 females,average age 66.5 years) who underwent GFR measurement with 99TcmDTPA and serum cystatin C and SCr tests were included in this retrospective study.GFR,serum cystatin C and SCr concentrations were determined by the Gates method (measured GFR),the particle-enhanced turbidimetric immunoassay,and the Jaffe method,respectively.Eight different equations (6 equations based on the serum cystatin C,and the other 2 based on SCr) were used to estimate GFR values,and the results were compared with that of the Gates method.Patients were divided into different groups according to the measured GFR (normalized to body surface area,1.73 m-2):normal renal function,mild,moderate or severe renal impairment groups.One-way analysis of variance and the least significant difference t test were used to compare the estimated GFR,andx2 test was used to compare the diagnostic efficiencies of different GFR estimation equations.Results Among 198 patients,159 cases were with renal impairment (78 mild,58 moderate,23 severe),and the other 39 cases were with normal renal function.For patients with moderate or severe renal impairment,the estimated GFR calculated by the Tan formula was not different from the measured GFR (severe:(20.7±7.4) ml · min-1 vs (19.9±8.2) ml · min-1; F=6.75,t＜1.05; moderate:(42.1±14.4) ml· min-1 vs (46.8±9.2) ml· min-1; F=10.49,t＜1.63; both P＞0.05),and it had the least error compared with the measured GFR (severe:(12.3±7.0) ％ ; moderate:(17.9± 13.0) ％).For the patients with mild renal impairment and normal renal function,the estimated GFR calculated by the Tan formula was not valuable.For the diagnosis of renal impairment,the sensitivity and accuracy of the modification of diet in renal disease (MDRD) formula were 66.0％(105/159) and 71.2％(141/198),respectively,and those of the chronic kidney disease-epidemiology collaboration (CKD-EPI) formula were 70.4％ (112/ 159) and 73.7％(146/198),respectively.The sensitivities and accuracies of the cystatin C-based formulas (≥83.6％ (133/159) and ≥79.3％(157/198),respectively) were higher than those of MDRD formula and CKD-EPI formula (x2 ≥23.50,all P＜0.01).For the diagnosis of chronic kidney disease (including 81 patients with moderate and severe renal impairment),the sensitivities of cystatin C-based prediction equations (≥ 86.4％ (70/81)) were higher than those of the MDRD formula and the CKD-EPI formula (76.5％ (62/81),79.0％ (64/81)),but the accuracies were slightly lower (Tan formula:80.3％ (159/198),x2≥ 56.42,all P＜0.05).Conclusion The Tan formula may be more suitable for the GFR estimation than the MDRD formula and CKD-EPI formula in the patients with severe or moderate renal impairment (serum cystatin C≥ 1.55 mg/L),but it may not be reliable for the patients with mild renal impairment and normal renal function.

As a kind of pesticide, fungicide and preservative, PCP had been used extensively in industry, agriculture and domesticity throughout the world. PCP contamination is generally associated with sediments or soil, it can also concentrate in organism. Regarding the character of high toxicity, long persistence and difficult to degrade, PCP has become a kind of conspicuous environmental pollutant because of widely use and inappropriate disposal. In the contaminated area, PCP can be detected in the water, soil and the body of organisms. PCP can affect human health through directly exposure or through food chain. The absorbed PCP can be stored in liver, kidney and fat,it can also increase the incidence rate of tumork, disturb the endocrine system, affect immune function,inhibit reproduction and development. PCP not only has a direct impairment on human body but also shows a potential impact in genetics.

AIM To explore the roles of vascular endothelial growth factor (VEGF) in hypoxia pulmonary hypertension and effects of suramin. METHODS Thirty SD rats were randomly divided into normal control group (N), hypoxia hypercapnia group(F), hypoxia hypercapnia +suramin group (S). The levels of VEGF in serum and in lung tissue were measured by ELISA, the ultrastructure of pulmonary arterioles was observed by electron microscopy, the expression of VEGF was observed by immunohistochemistry, the expression of VEGFmRNA was observed by in situ hybirdization. RESULTS ①Mean pulmonary arterial pressure(mPAP), weight ratio of RV to LV+S, the leves of VEGF in serum and in lung tissue of group F were significantly higher than that of group N and group S (P

AIM: To observe the expression of proteins induced by whole-body irradiation in mice with low dose X-rays and their biological activity. METHODS: To analyse the expression of protein induced by whole-body irradiation with 75mGy X-rays and sham-irradiation, the method of gel filtration with Sephadex G-100 and HPLC were used. The biological activity of protein expressed in thymocytes was observed by mouse splenocyte proliferation and chromosome aberration of human peripheral blood lymphocytes. RESULTS: HPLC analysis showed that there was a marked decrease in expression of 24.5 kD protein molecule in the fractions of thymocyte extract in comparison with the corresponding fractions from the sham-irradiated control mice. These protein fractions from the thymocytes of the irradiated mice at the concentration of 6.25 mg/L showed both inhibitory effect on normal T cell proliferation and protective effect on the chromosome damage induced by high dose radiation. CONCLUSION: Down regulation of 24.5 kD protein molecule expression may have implications in the mechanism of immunoenhancement and cytogenetic adaptive response induced by low dose radiation.

Objective　To manufacture adriamycin-porous tricalcium phosphate (A-PTCP) ceramic drug delivery system (DDS)as a possible method for bone defect treatment after bone tumor operation. Methods　A-PTCP DDS was made from putting adriamycin into PTCP. Thirty rabbits were divided randomly into group A(24 rabbits) and group B(6 rabbits). A-PTCP was implanted in the greater trochanter of the right femur in group A. Adriamycin were injected into veins in group B. Muscle around A-PTCP and plasma were taken out at different period. Adriamycin concentrations in muscle and plasma were measured by high performance liquid chromatography (HPLC). Results　A-PTCP could gradually release adriamycin over 10 weeks. Adriamycin concentrations in the muscle were higher than that in plasma. Conclusion　A-PTCP may be a new method for repairing bone defects after bone tumor operation.