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.

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 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.

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.

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.

OBJECTIVE To investigate the efficacy and feasibility of endoscopic surgery to manage the tumors involving naso-ethmoidal complex and anterior skull base.METHODS 19 cases of tumors involving nasal cavity,ethmoid sinus and anterior skull base were summarized retrospectively, including 3 cases of ethmoidal squamous carcinoma, 3 cases of esthesioneuroblastoma,3 cases of melanoma,1 case of plasmocytoma,2 cases of ethmoidal adenocarcinoma,4 cases of adenoid cystic carcinoma,2 cases of meningioma.RESULTS Tumors were totally removed in 18 cases and subtotally removed in 1 case,all the cases were followed up for 1 to 3 years.Of the 3 cases of melanoma,1 case died of brain metastases 1 year after surgery,1 case recurred 8 months after surgery,1 case of adenoid cystic carcinoma recurred 17 months after surgery.No recurrence were found in the rest 16 cases during 1-3 years of following-up.CONCLUSION Endoscopic surgery is a effective method for malignant tumors localized in nasal cavity and ethmoid sinus and benign anterior skull base tumors involving nasal cavity and sinuses.Indication must be carefully selected for malignant tumors and postoperative radiotherapy should be advised.