China ; epidemiology ; Data Collection ; standards ; Humans ; Occupational Diseases ; chemically induced ; epidemiology ; Occupational Exposure ; statistics & numerical data ; Occupational Health ; statistics & numerical data ; Occupational Medicine ; legislation & jurisprudence

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.
Antineoplastic Agents ; pharmacology ; Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Humans ; Oligopeptides ; biosynthesis ; pharmacology ; Pichia ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; pharmacology

Antiviral Agents ; therapeutic use ; Hepatitis C ; drug therapy ; Humans ; Liver Cirrhosis ; drug therapy ; virology

One of the key contents in tissue engineering trachea replacement research is the scaffold selection.This review summarizes the latest original literatures and investigations about electrospun technique as well as recent progress.To discuss the advantages and disadvantages of natural,synthetic and combined electrospun scaffolds,the versatility in material choosing and production methods is the unique superiority.For specific experimental or clinical objects,the further research is to choose a suitable polymer,to improve surface modification techniques and to control the dimension and arrangement of the fibrous structure of electrospun fibers,which can offer versatility and tissue specificity,and therefore provide a bright prospect for tissue engineering trachea.

Bile Duct Neoplasms ; pathology ; surgery ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; pathology ; surgery ; Humans

Objective To explore the effects of the therapeutic effects of interventional heated chemotherapy( IHC) for moderate and advanced huge primary hepatocellular carcinoma and compare with transcatheter arterial chemoembolization( TACE) . Methods The 68 pa-tients with moderate and advanced huge primary hepatocellular carcinoma accepted the treatment of IHC were considered as the observation group,and 32 patients managed TACE were regarded as the control group. The therapeutic effects,living quality,secondary surgery rates,The reduced level of serum AFP,survival rate,and toxic side effects of 2 groups were contrasted. Results The effective rate of observation group was obviously higher than that of control group (79. 4% vs 56. 3%,P <0. 05),and secondary surgery rates was increased(70. 6% vs 43. 8%,P<0. 01). The ratio of increased Karnofsky score was more than 20 in observation group,which was obviously higher than that of control group(35. 3% vs 12. 5%,P<0. 01). T The reduced level of serum AFP was more than 50% in observation group,which was higher than that of control group (52. 9% vs 25. 0%,P<0. 01). The survival rate of 12,18,24 months after treatment in control group was in-creased than that in control group individually (P<0. 01). Conclusion IHC is effective for huge primary hepatocellular carcinoma and in-creases the secondary surgery rates,which is one of the safe and effective methods for huge primary hepatocellular carcinoma.

Animals ; Diet, High-Fat ; Glucagon ; blood ; Glucose ; metabolism ; Hormones ; blood ; Insulin ; blood ; Leptin ; blood ; Lipid Metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effect of thalidomide on the proliferation of as well as the expression and secretion of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) by the human keratinocyte cell line HaCaT.Methods Cultured HaCaT cells were divided into several groups to be treated with dimethyl sulfoxide (negative control group) and various concentrations (0.01nmol/L-100 μmol/L) of thalidomide (experimental groups) respectively for 20 to 24 hours.Subsequently,water soluble tetrazolium-1(WST-1) assay was performed to estimate cellular proliferation,real time quantitative PCR to detect the mRNA expression of VEGF and TNF-α in HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to quantify the protein expressions of VEGF and TNF-α in the culture supernatants of HaCaT cells.Statistical analysis was done by one-way analysis of variance with least significant difference post hoc test.Results The survival rate of HaCaT cells was 74.3％,82.9％ and 90.8％ after 24-hour treatment with thalidomide of 100,10 and 1 μmol/L respectively,significantly lower than that in the negative control group (100％,all P ＜0.01).A significant decrease was induced in the mRNA expression (0.439-to 0.634-fold change,all P ＜0.01) and supematant level ((0.587-to 0.923-fold change,P ＜0.05) of VEGF in HaCaT cells by thalidomide of 0.01-100 nmol/L,as well as in the mRNA expression (0.493-to 0.587-fold change,P ＜0.05) and supernatant level (0.408-to 0.617-fold change,P ＜0.01) of TNF-α by thalidomide of 0.1-100 nmol/L.Conclusion Within a certain range of concentration,thalidomide could suppress the proliferation of,as well as the expression and secretion of VEGF and TNF-α by,HaCaT cells.

Animals ; Hypertension ; metabolism ; physiopathology ; Male ; Myocardium ; metabolism ; Peptidyl-Dipeptidase A ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism

Background and purpose:Neoadjuvant chemotherapy is an excellent model for evaluation of predictive parameters.The goal of predictive parameters is to select patients who would most likely benefit from neoadjuvant chemotherapy.The objective of this study was to investigate the predictive value of biological markers for the patients' response to neoadjuvant anthracycline combined with taxanes chemotherapy.Methods:We investigated 160 patients with breast cancer who underwent 4 cycles of neoadjuvant anthracycline combined with taxanes chemotherapy,retrospectively.The expression of estrogen receptors(ER),progesterone receptors(PgR),Her2,Topo-Ⅱ and Ki-67 proteins were detected by immunohistochemical assay in core-needle biopsy specimens.The associations between biological markers as well as clinical and pathologic responses were analyzed.Results:The overall clinical response was 85%,including 28.8%(46/160) with clinical complete response(cCR) and 56.3%(90/160) with clinical partial response.The pathological complete response(pCR) was 14.4%.In the univariate analysis,absence of ER,PgR expression and over-expression of Her-2 were predictive of the cCR and pCR(P