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 evaluate the impact of epidural anesthesia with levobupivacaine combined with general anesthesia on colon surgery. Methods Sixty patients undergoing elective radical procedure for colon carcinoma were randomLy divided into four groups: saline group (group S), 0.125% levobupivacaine group (group L1), 0.25% levobupivacaine group (group L2), and 0.5% levobupivacaine group (group L3). Group S received normal saline of 10 mL epidurally and then infusion of 5 mL·h-1 until the procedure was finished; groups L1, L2, and L3 received levobupivacaine instead. Anesthetic induction was performed after epidural puncture. Mean blood pressure and heart rate were recorded at 8 time points including 5 min after entering into the operation room, 1 min after intubation, skin incision, abdominal exploration, 1 h after skin incision, completion of operation, extubation, and leaving PACU; meanwhile blood glucose and cortisol were detected, anesthesia time, time to PACU stay, bleeding, transfusion volume, adverse reaction, and doses of propofol, remifentanil, ephedrine, and fentanyl were noted. Results Time to PACU stay was longer in S group than in other 3 groups. Doses of remifentanil and fentanyl were larger in L1 group than in L2 group and L3 group. Ephedrine dose in L3 group was larger than in other 3 groups. Blood sugar in L1 group was higher than L2 group and L3 group. Cortisol in S group was higher than in other 3 group. Cortisol in L1 group was higher than in L3 group. The number of patients with hypotension was greater in L3 group than other 3 groups. Conclusions Continue epidural infusion of 0.25%levobupivacaine can reduce stress response and opioid uses, shorten PACU stay, whereas it does not increase use of ephedrine.

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.

Adolescent ; Child, Preschool ; Eosinophilia ; complications ; Female ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; complications

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

Adult ; Humans ; Male ; Occupational Diseases ; Pulmonary Aspergillosis

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.

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.