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

Breast Neoplasms ; classification ; genetics ; metabolism ; pathology ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Homeodomain Proteins ; metabolism ; Humans ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Oligonucleotide Array Sequence Analysis ; methods ; Receptors, Interleukin ; metabolism ; Signal Transduction ; Survival Rate

Oncology is extensive in contents,covering a wide range of organs,systems and clinical specialties.Here,we discuss the feasibility and necessity of flipping the classroom teaching through the introduction of Oncology and through the implementation of flip classroom teaching for oncology graduate students,this paper analyzes the evaluation of the classroom teaching by the teachers and students,and compares the assessment results of students under different teaching methods.The results show that the flipped class can promote students' initiative learning,promote students' classroom participation,and help students to internalize and consolidate their knowledge of oncology in the theoretical teaching of graduate oncology.

Objective To observe the effects of exercise on serum adiponectin and adiponectin receptor (AdipoR) level in skeletal muscle of insulin-resistant rats. Methods A total of 30 healthy male rats were randomly divided into a control group ( NC, n = 8) and a high-fat group ( HF, n = 22), fed with normal chow and high fat diet, respectively. Eighteen weeks later, the high-fat group was randomly divided into a high-fat diet control group (HC, n = 10) and an exercise group (HE, n = 12). The HC and HE group were continually fed with high fat diet, while the HE group was administered with swimming training for 6 weeks in addition at the same time. After 24 weeks, the insulin sensitivity index was calculated, and serum adiponectin level was detected by using ELISA. The expressions of AdipoR mRNA in skeletal muscle were detected with real-time fluorescence quantitative polymerase chain reaction (PCR). Results After 18 weeks, compared to NC group, the insulin sensitivity index of HF group decreased significantly. It suggested that insulin resistance appeared in HF group. Twenty-four weeks later, compared to NC group, the ISI of HC group was significantly decreased, meanwhile the level of serum adiponectin, expression of AdipoR1 and AdipoR2 mRNA in skeletal muscle of HC group were 71.9% , 59.9% and 69.2% of those of the NC group, respectively; compared to HC group, the ISI was increased significantly by exercise, meanwhile the expression of AdipoR1 mRNA in skeletal muscle was significantly increased by 1.33 times, however the level of serum adiponectin and the expression of AdipoR2 mRNA in skeletal muscle were not altered in HE group. Conclusion Six weeks of exercise improves insulin sensitivity through increasing the expression of AdipoRI mRNA in skeletal muscle.

Adult ; Humans ; Male ; Occupational Diseases ; Pulmonary Aspergillosis

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carbon Monoxide Poisoning ; epidemiology ; therapy ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Inpatients ; statistics & numerical data ; Male ; Middle Aged ; Young Adult

Objective To investigate the knowledge of chronic heart failure (CHF) and influencing factors in general practitioners (GP) in Beijing.Methods A self-designed questionnaire contained total 28 items,including basic knowledge of CHF,non-drug management,drug management and other management ; clinical cases were used to test clinical ability in 7 items.The questionnaire survey was conducted among GPs who participated in continuing education courses from 16 counties/districts in Beijing during January to June 2013.Results Total 720 questionnaires were distributed and 657 valid questionnaires were returned with a recovery rate of 91.3％.Total scores was 60.6.Scores of basic knowledge,non-drug management,drug management and other management was 63.1,76.9,44.0 and 56.9,respectively.There were statistical differences in scores of basic knowledge,non-drug management,drug management and other management between GPs with different diploma (F value:36.8,5.8,21.6,12.2,respectively; P ＜0.01) ; there were significant differences in scores of basic knowledge and drug management among GPs with different working years (F value:15.1 and 17.4,respectively ; P ＜ 0.01) ; there was significant difference in scores of drug management among GPs with different professional title (F =7.69,P ＜ 0.01).Only for GPs with junior college diploma,the scores of basic knowledge and drug management in GPs with working ≥20 y were higher than those working ＜ 20 y(P ＜ 0.01).The accuracy of clinical ability in GPs with junior college diploma,undergraduate diploma and post-undergraduate diploma was 39.6％,41.6％,41.8％ (P ＞ 0.05).Conclusions The knowledge of CHF is less desirable in GPs of Beijing,so that measures should be taken to improve the GP's knowledge of CHF.

Objective:To analyze the distribution of UGT1A1 gene polymorphisms in Chinese Han patients with extensive-disease small cell lung cancer(ED-SCLC),and evaluate the correlation between UGT1A1 gene polymorphisms and toxicity and efficacy of irino-tecan(CPT-11) based regimen in the patients with ED-SCLC. Methods: The analysis of UGT1A1?28 and UGT1A1?6 gene poly-morphisms was performed in 67 patients with ED-SCLC admitted in our hospital from June 2011 to January 2013. The 67 cases with ED-SCLC treated with irinotecan(CPT-11) based regimen were enrolled to observe the adverse events and efficacy during the chemo-therapy, including objective responserate rate ( ORR) , progression free survival ( PFS) and overall survival ( OS) . The incidence of different genotypes was compared. Results:The distribution of UGT1A1 genotypes in the 67 patients was follows:UGT1A1?28 wild-type (WT) genotype TA6/6 (56, 83. 6%), heterozygous genotype TA6/7 (11, 16. 4%);UGT1A1?6 wild-type (WT) genotype G/G (45,67. 2%), heterozygous genotype G/A (22,32. 8%). No significant difference of PFS and OS was observed between the differ-ent genotypes. The incidence of grade 3 and 4 delayed diarrhea and neutropenia in the patients carrying UGT1A1?6 G/A was higher than that in those with WT genotype (36. 4% vs. 6. 6%, P<0. 05;27. 2% vs. 4. 4%, P<0. 05, respectively). The incidence of grade 3 and 4 thrombocytopenia in the patients carrying UGT1A1?28 TA6/7 was higher than that in those with WT genotype (27. 2%vs. 1. 8%, P<0. 05). The patients simultaneously carrying UGT1A1?28 TA6/7 and UGT1A1?6 G/A were prone to suffering 3 and 4 delayed diarrhea and neutropenia. Conclusion: UGT1A1 polymorphisms may predict the adverse events of CPT-11 in ED-SCLC, while can not predict the efficacy of CPT-11.

The recombination clones contained CFP, LTB-ST foreign gene were screening by PCR using individual bacterial colonies as template, the aimed band was amplified from positive clones, the result was as well as plasmid PCR. The selecting of agrobacterium transformed with recombination plasmid could also use this method of PCR screening of individual bacterial colonies. The result of individual bacterial colonies PCR was as well as that of PCR using bacterial solution as template. It showed that the method individual bacterial colonies PCR was an efficient, easy one that characterized recombination clones.

AIM: To examine the change of serum tumor necrosis factor-? (TNF-?), nitric oxide (NO) in patient with congestive heart failure (CHF) and the effect of angiotensin Ⅱ (AngⅡ), valsartan on TNF-? and NO production in culture peripheral blood mononuclear cells (PBMC), to assess the relationship between the renin-angiotensin system and cytokines. METHODS: Venous blood of both healthy volunteers (n=12) and patients with CHF (n=16) were collected. Serum TNF-? and NO were examined. Peripheral blood mononuclear cells (PBMC) were obtained from both the control and the patients groups and cultured with AngⅡ at concentrations of 0, 0.01, 0.1, 1 ?mol/L, respectively. AngⅡ at concentration of 0.1 ?mol/L combined with 0.1 ?mol/L of valsartan was also used. After 24 h incubation, the contents of TNF-? and NO in the culture supernatants were measured. RESULTS: Serum TNF-? and NO production in CHF group were significantly higher than that in control group (P0.05) were observed. AngⅡ stimulated TNF-? and NO release from PBMC of patients with CHF and normal person, which was inhibited by valsartan. CONCLUSIONS: AngⅡ obviously increases TNF-? and NO production from PBMC, which indicates there is relationship between the renin-angiotensin system and TNF-?, NO. The fact that valsartan inhibits TNF-? production may be one of the mechanisms in treating CHF.