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

The brain structural imaging changes in patients with post-traumatic stress disorders (PTSD) and the related research progress are reviewed. It is elucidated that the volume decrease and the density reductions of gray and white matters in PTSD patients are mainly manifested as the imaging changes of the hippocampus, callus, and anterior cingulate cortex (ACC). The imaging change of callus is characteristic in PTSD patients. The image structure analyses of the white matter and gray matter are the current frontiers and strategic trends of PTSD-related research. Meanwhile, it is pointed out that the relationship between PTSD and brain structure imaging is a problem needs to be urgently solved.

Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Cyclophosphamide ; administration & dosage ; Doxorubicin ; administration & dosage ; Humans ; Lymphoma, B-Cell ; classification ; drug therapy ; therapy ; Prednisone ; administration & dosage ; Stem Cell Transplantation ; Vincristine ; administration & dosage

Based on vector auto-regression (VAR) model, this paper takes advantage of Granger causality test, variance decomposition and impulse response analysis techniques to carry out a comprehensive study of the factors influencing the price of Chinese herbal, including herbal cultivation costs, acreage, natural disasters, the residents' needs and inflation. The study found that there is Granger causality relationship between inflation and herbal prices, cultivation costs and herbal prices. And in the total variance analysis of Chinese herbal and medicine price index, the largest contribution to it is from its own fluctuations, followed by the cultivation costs and inflation.
Agriculture ; economics ; Commerce ; Costs and Cost Analysis ; Drugs, Chinese Herbal ; economics ; Humans ; Models, Statistical ; Plants, Medicinal ; growth & development ; Regression Analysis

Cerebral Palsy ; mortality ; pathology ; physiopathology ; Child ; Humans ; Prognosis ; Survival Analysis ; Survival Rate ; Time Factors ; Walking

1-Alkyl-2-acetylglycerophosphocholine Esterase ; blood ; genetics ; Adolescent ; Asthma ; blood ; enzymology ; genetics ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Enzymologic ; Genotype ; Humans ; Infant ; Male ; Mutation ; Polymerase Chain Reaction

China ; Haplopappus ; Humans ; Phytotherapy ; Sesquiterpenes

Objective To evaluate the feasibility of contrast material on image quality of coronary CT angiography (CCTA)by using double injection technology and iterative reconstruction.Methods 120 patients with suspected coronary heart disease who underwent CCTA were randomly divided into two groups.Then,60 patients with 30 kg/m2>BMI≥25 kg/m2 were averagely divided into A1 and B1 groups,and other 60 ones with BMI<25 kg/m2 were also averagely divided into A2 and B2 groups.The patients in group A were scanned with 120 kV tube output and iolromide 350 mg I/mL,and the images were reconstructed using filtered back projection (FBP).Meanwhile,the patients in group B were scanned with 80 kV or 100 kV tube output and 245 mg I/mL or 280 mg I/mL of contrast media (BMI≥25 kg/m2 )using double injection technology,and the images were reconstructed using adaptive iterative dose reduction 3D (AIDR 3D).The effective radiation dose (ED)of each patient was also calculated.CT attenuation of the main proximal vessels (ascending or descending aorta,pulmonary artery,RCA,LM,LAD and LCX)and the obj ective image quality (SD,CNR and SNR)were measured,and the subj ective evaluation was also assessed by an experienced radiologist.Results There was significant differences in mean image quality scores,SD,SNR and CNR of ascending aorta between group A1 and B1(P<0.05),but no difference in the SI of values (P>0.05).There was no significant difference in mean image quality scores between group A2 and B2 (P>0.05), however,there were significant differences in the SI,SD of values and SNR,CNR of ascending aorta (P<0.05).The values of BMI were negatively related to the values of CT in ascending aorta between two subgroups (r=-0.66 and -0.441,P<0.05).The values of weight were not related to the values of CT in ascending aorta (P>0.05).There was significant difference in effective dose between subgroups (P<0.05).Conclusion The method with a combination of iterative reconstruction and a contrast material of 245 mg I/mL using double inj ection technology can improve the contrast enhancement without impairing image quality.

Cerebral Palsy ; complications ; Child ; Child, Preschool ; Epilepsy ; etiology ; Humans

Objective To explore the relationship between the expression of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) in gestational trophoblastie diseases and its invasiveness.Methods The expression and distribution of MMP-7,MMP-9,TIMP-1 and TIMP-2 were determined by immunohistochemistry PV-6000 in 49 cases of hydatidiform mole (eight of them with malignant transformation),six cases of invasive hydatidiform mole,two cases of choriocarcinoma and 18 cases with normal villi in early pregnancy.Results The expression of MMP-7,MMP-9,TIMP-2 and MMP- 9/TIMP-1 was significantly higher in choriocareinoma,invasive mole and malignant transformed mole than that in normal villi in early pregnancy and non-malignant transformed mole (F=10.950,7.760,7.304, 3.442;P