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

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

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.

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

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

Objective To systematically review the efficacy and tolerance of levofloxacin-based rescue regimens for H.pylori eradication failures.Methods A search of Medline,Embase,Cinahl, CBM-disc was performed.Randomized controlled trials comparing levofloxacin-based triple therapy with bismuth-based quadruple therapy were selected for meta-analysis.Assessment of study quality and ex- tracting data to calculate eradication rate and side effect rate.Results Seven randomized controlled stud- ies met the inclusion and exclusion criteria were recruited.Ten-day levofloxacin-based triple therapy was more effective(86.9% vs 61.8% P

Abstract? AIM: To analyze the ocular manifestations of meningiomas in the trigone of the lateral ventricle, discuss the relevant factors of visual impairment in these patients and things need attention clinically.?METHODS:Retrospectively study on the clinical data of 90 eyes in 45 patients diagnosed of trigonal meningiomas treated at Beijing Tian Tan Hospital from October 2011 to October 2015.Preoperative examinations including visual acuity, optic disc findings, visual field, size of tumors and other change in MRI were analyzed.?RESULTS: Patients'age was 12-68 years old ( mean 41.7 ±13.7 years ). Male/female ratio was 1 ∶4.6. Decreased visual acuity occurred in 18 eyes.Optic disc edema was found in 24 eyes and optic disc pale in 6 eyes. Fourty -seven eyes had visual field defect, mostly homonymous hemianopia or defect. The maximum diameter of tumors was 2.1-9.6cm (4.8±1.7cm).Range of tumor volume was 3.02-193.2cm3(48.3±47.8cm3).A positive correlation of preoperative visual field defect was found with tumor volume, tumor maximum diameter, and brain midline shift respectively. While the preoperative visual field defect was not found any correlation with age, gender, course, and the enlargement of the ventricle and the edema of the tissue around the tumors. After Mann-Whitney U test, the differences on tumor volume, the maximum diameter of tumors, and brain midline shift between the two groups were significant.? CONCLUSION: Patients with trigonal meningiomas often have ocular signs and symptoms.The major reason of visual field defect is the damage of optic radiation around the tumor.The specific position of injured optic radiation determines the type and extent of visual field defect.Both ophthalmologist and neurosurgeon should pay attention to tumors nearby posterior visual pathway. Doing neurophthalmology examinations for these patients and realizing the position between the tumor and posterior visual pathway will be helpful and necessary in surgical planning.

Objective To explore the mobile changes of nitric oxide(NO) and the relationship between nitric oxide and glomerular hyperfiltration in experimental diabetic rats.Methods Experimental diabetes mellitus(DM) was induced in rats with streptozotocin(STZ).The levels of NO and NOS in renal tissue homogenate were assayed after establishment of diabetesat the 4 th,8 th,12 th week.At the same time, renal morphology in diabetic rats was examined by light microscope and image of computer.Results The contents of NO and NOS in renal homegenates were evidently increased at 4 th week,and decreased gradually from 8 th week(P

Objective To study the changes and significance of Bid gene expression in hypoxic-ischemic brain damage (HIBD) in neonatal rats.Methods The animal models of HIBD were made. Total RNA from ipsilateral cerebral hemisph ere was extracted.RT-PCR was used to evaluate the levels of Bid gene expression at different time points after hypoxic-ischemic. Results The levels of Bid mRNA were higher in HIBD rats than those in normal rats.The ex pression of Bid mRNA in ipsilateral hemiphere following cerebral hypoxic-ischem ic began to increase at 6 h, increased to the highest level at 24 h, and then gr adually returned.Conclusion The upregulation of Bid mRNA can play a role in the induction of apoptosis fol lowing cerebral hypoxic-ischemic. J Appl Clin Pediatr,2005,20(2):121-122