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

Vascular endothelial growth factor(VEGF) is one of the best characterized angiogenic regulators which has many effects in promoting endotheliocyte proliferation and differentiation,increasing the microvascular permeability,inducing the angiogenesis,triggering the growth,survival and migration of tumor cells by combining its specificity receptor (VEGFR).Dysregulation of VEGF expression and signaling pathways therefore plays a pivotal role in the pathogenesis and clinical features of hematologic malignancies,direct and indirect targeting of VEGF and its receptors therefore may provide a potent novel therapeutic approach to overcome bone marrow angiogenesis and multidrug resis-tance thereby improve patient outcome.Recent years,a novel VEGF blockade system using RNA interference attracts more and more people's attention.The small interfering RNA (siRNA) targeting VEGF/VEGFR can completely inhibits the expression of VEGF and induce the silence of corresponding genes.

0.05). Compared with the healthy neonates, the concentration of blood MDA and ROS was significantly higher, and that of blood HB and SOD was significantly lower in both groups(P

The sensor of naringin(NG), a non-electroactive substance, was prepared based on the molecular imprinting technique.Using cyclic voltammetry technique (scan rate is 100 mV/s), the naringin imprinting sensitive film, poly-o-aminophenol was coated on the surface of a graphite electrode in the presence of naringin which was considered as the template, and characterized by SEM and X-ray reflective spectrophotometry (XRR).Using K_3Fe(CN)_6 as an electroactive marker, the electrochemical properties of the NG sensor were investigated by CV, electrochemical impedance spectroscopy (EIS), differential pulse voltammmetric and chronoamperometric.The results showed that the imprinted electrode was significantly different from the non imprinted electrode in morphologies and electrochemical properties, and a linear relationship between the peak current and the naringin concentration was found in the range of 6.0 × 10 ~(-5)-1.4 × 10 ~(-4) mol/L with a detec tion limit of 1.6 × 10 ~(-5) mol/L.Moreover, the imprinted electrode exhibited a good selectivity and rapid response to the naringin template molecules, as well as an excellent reproducibility(RSD = 1.8 %, n=5).

Apoptosis ; Inhibitor of Apoptosis Proteins ; Myocytes, Cardiac ; cytology

Congenital Abnormalities ; diagnosis ; therapy ; Ear, External ; abnormalities ; Ear, Middle ; abnormalities ; Humans

Objective To compare the difference of early trauma experience between early-onset and late-onset obsessive-compulsive disorder(OCD),and the kinds of early trauma in the cause of OCD. Methods Three hundred and twenty-six patients with obsessive-compulsive disorder and 480 healthy con-trols were enrolled. Early Trauma Inventory-Short Form ( ETI-SF) was applied to assess the early trauma ex-perience,compare the difference of early trauma experience between early-onset ( n=138) and late-onset ( n=188) OCD,and the kinds of early trauma in the cause of OCD. Results There were significant differences in the early trauma experience between obsessive-compulsive disorder group and control group((3.90±3.61) vs (1.88±2.61), P<0.01).Scores of physical((1.12±1.38) vs (0.71±1.23), P=0.001),emotional((1.58± 1.71) vs (0.42±1.01), P<0.01),and sexual abuse((0.27±0.59) vs (0.09±0.36), P=0.001),and a gener-al traumatic experience((0.94±1.17) vs (0.66±1.09), P<0.01) were significant difference between patients and controls. Except sexual abuse((0.30±0.66) vs (0.24±0.67), P=0.42),there were significant differ-ences in the early trauma between early-onset and late-onset OCD((5.12±3.58) vs (3.01±3.38), P<0.01), especially emotional abuse((2.18±1.76) vs (1.12±1.53), P<0.01).247(75.8%) OCD patients and 79(57. 9%) healthy controls experienced early trauma(χ2=21.48, P<0.01). Experiencing one kind of early trau-ma,the prevalence of OCD was 39.1%,two kind was 61.0%,three kind of early trauma was 65.8%,and ex-periencing four or more kind of early trauma,the prevalence of OCD was up to 84.4%. Conclusion OCD patients have much more early trauma,especially early-onset OCD. Experiencing more kind of early trauma, and the prevalence of OCD higher may associated with the development of obsessive-compulsive disorder. It is important to consider the role of childhood trauma in the prevention and treatment of OCD.

Objective To examine the expression of neuron-specific enolase (NSE) in patients with remission diffuse large B-cell lymphoma (DLBCL),and to explone its prognostic value.Methods Serum NSE values were mesured by electmchemilumineseence immunoassay in 20 health donors and 40 DLBCL patients reached remission after 6-8 cycles of chemotherapy after initial diagnosis.Results Positive expressions rate of NSE in remission DLBCL patients was 50 ％,and was 10 ％ in health donors (P ＜ 0.01).In remission DLBCL patients,there was significant difference in the 2-year PFS rate between in NSE-positive group and negative group (6.6 ％ vs 20.9％,P ＜ 0.001).Serum NSE was an independent poor prognostic factor in remission DLBCL patients.Conclusion Serum NSE expressions is an independent poor prognostic factor of remission DLBCL and has significant clinical value.

Objective To study a novel method for the high-dose-rate brachytherapy (HDR) CT image to the intensity modulated radiation therapy (IMRT) CT image deformable image registration and dose accumulation.Methods The applicator in the HDR CT image is first segmented and removed,then a deflation step is performed on the applicator-free HDR CT image by solving the Navier-Stokes equation.Demons algorithm is utilized to register the deflated HDR CT image to the IMRT CT image,along with the HDR dose.The deformed HDR dose is then added on the IMRT dose and yield the final accumulated dose.Results The HDR CT image and IMRT CT image,as well as the corresponding dose distribution,from five cervical cancer patients are used for evaluation of the proposed algorithm,the results show that the proposed method can effectively get rid of the influence of the applicator and produce an accurate accumulated dose.Conclusions Dose accumulation and supervision is an important step in adaptive radiotherapy for accurate dose delivery and treatment plan re-optimization.The proposed method in this study can effectively accumulate the HDR dose to the IMRT dose domain,and the accuracy is proved to be sufficient for clinical needs.

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.