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

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

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.

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.

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

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

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

Objective To evaluate left ventricular systolic synchrony in patients with Ⅲ grade atrio-ventricular block(Ⅲ grade AVB) after pacemaker implantation by real-time three-dimensional echocardiography. Methods Methods Twelve patients with Ⅲ grade AVB implanted by VVI pacemaker, seven patients with Ⅲ grade AVB implanted by DDD pacemaker, twenty subjects with normal LV function were examined by Philips iE33 with a X3-1 probe. Results The minimal excursion in patients with Ⅲ grade AVB implanted by VVI pacemaker and DDD pacemaker was less than that in subjects with normal LV function (P <0.05) ,and two "bull eyes" displayed that some regional systolic timing delayed or ahead, but also some regional systolic excursion hypokinesis or dykinesis. Parameter index [including Tmsv 16-SD, Tmsv 12-SD, Tmsv 6-SD, Tmsv 16-Dif, Tmsv 12-Dif, Tmsv 6-Dif, Tmsv 16-SD (%), Tmsv 12-SD (%), Tmsv 6-SD (%), Tmsv 16-Dif (%), Tmsv 12-Dif (%), Tmsv 6-Dif(%)] of 17 segment time to minimal systolic volume in patients with Ⅲ grade AVB implanted by VVI pacemaker and DDD pacemaker was more than that in subjects with normal LV function (all P < 0.05). Conclusions Real-time three-dimensional echocardiography can rapidly evaluate left ventricular systolic synchrony in patients with Ⅲ grade AVB implanted by DDD pacemaker and VVI pacemaker.

Objective To evaluate the value of fine needle aspiration (FNA) and core needle biopsy (CNB) in highly suspicious breast malignant lesions in terms of diagnostic accuracy,complication rate and cost-effectiveness.Methods We retrospectively reviewed records of patients with imaging diagnosis of BI-RADS 4c or 5 categories who have undergone either FNA or CNB under ultrasound guidance in Peking University People's Hospital from 2012 to 2014.Sensitivity,specificity,diagnostic accuracy,positive predictive,negative predictive,false positive rate,false negative rate,accuracy rate and unsatisfactory rate (non-diagnostic rate) were calculated and compared between FNAC and CNB.The complication of the procedures,operation time and cost were assessed.Results Among 638 consecutive cases,273 of them underwent FNA,and 365 underwent CNB.The accuracy rate of FNA and CNB were 99％ and 97.2％.The sensitivity,specificity,diagnostic accuracy,positive predictive,false positive rate and false negative rate were similar between the two groups.The unsatisfactory rate of FNA was significantly higher than that of CNB (25.3％ vs.10.9％,x2 =22.59,P =0.000).There was not severe complication in either groups,while subcutaneous ecchymosis and hematoma were more common in FNA group than in CNB group.The operation time of FNA was shorter than that of CNB (4.8 ± 1.3 min vs.15.5-± 1.7 min,P =0.000).The waiting time for final pathological report was shorter in patients undergoing FNA (1 vs.3 days).An estimated cost for FNA was RMB 447.5 Yuan/case,whereas that was 995 Yuan/case for CNB.Conclusions FNA and CNB are both accurate and safe preoperative diagnostic procedures.FNA is a simple and cost-effective method.

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