The prospective evaluation of the transformation feasibility of the research achievements is an important portion of the evaluation of the medical research achievements.This study explored to set up a system with Delphi method for the evaluation guideline of the hlansformation feasibility of the medical research achievements.And it also offered some evaluating methods and theoretical guidance to carry out the transformation feasibility of the achievements in the medical scientific research.

Objective To investigate the feasibility of searching for proteins which interact with intracellular domain of hematopoietic growth factor receptors using yeast two-hybrid system. Methods RT-PCR method was performed to amplify the genes of intracellular domains of G-CSF receptor and EPO receptor in NFS-60 and BET-2 cells of mice. The genes were cloned into yeast expression plasmid pGBKT7 vector,and then transformed into yeast AH109. The yeast proteins were isolated and analyzed with Western blotting. Transcriptional activation was analyzed by the ?-galactosidase colony-lift filter assay. Results The intracellular domains of G-CSF receptor and EPO receptor genes were successfully cloned into pGBKT7 vector. The results of Western blotting assay showed that both proteins were expressed in the yeast cells. The ?-galactosidase colony-lift filter assay demonstrated that G-CSF receptor alone had no activity of transcriptional activation,while the EPO receptor alone could activate transcription. Conclusion The findings suggested that intracellular domain of G-CSF receptor could be used as a bait to find interacting proteins using yeast two-hybrid system,while that of the EPO receptor could not. Therefore the system could not be applied to all hematopoietic factor receptor.

Abstract Objective: To construct the Second half of CILP(C2) MBP fusion protein by sub-cloning technology. Methods: Recombinantfusion proteins, which contain the fragments within the C2 region(designated C2F1, C2F2 and C2F3) of the non-porcine nucleotide pyrophos-phohydrolase-homologous region of CILP, were prepared using pMAL-eHis vector. The recombinat genes are induced by different temperatures(22℃,30℃,37℃ ). Results: Expression using pMAL-eHis system can be induced chemically by adding IPTG. 37℃ temperature prmotes in-soluble inclusion-body formation,but 22℃ temperature can not induce the enough expression of recombinant protein. Onl 30℃ temperaturecan induce enough amount of soluble recombinant protein. The characers of fusion proteins that they carried 6 straight histidines, (His)6, at tbeC-terminus of multiple cloning sites for affinity purification were assessed by sodium dodecy1 sulfate-polyacylamide gel electrophoresis(SDS-PAGE) and Western blot. Nucleotide sequences of the insertion genes were confirmed by dideoxy sequencing. Conclusion: C2F1, C2F2,C2F3-MBP fnsion proteins were constructed successfully.These recombinant proteins may provide important roles in the future study on CILP.

Objective To study the correlation between Candida strains and vulvovaginal candidiasis. Methods Two groups of Candida albicans strains were chosen. Strains of group A and group B were isolated from vaginal discharge of normal women and of patients with candidal vaginitis respectively. The two methods, AMS (automated microbiological detection and identification system) and amino black stain, were applied to detect two phenotypic parameters, biochemical reaction of the strains and secretory capacity of the proteinases respectively, and analogous analysis was performed then. Results There was a significant difference in the levels of adonitol (P

Objective To discuss the survivorship of rat mesenchymal stem cells (rMSCs) and the expression of human brain-derived neurotrophic factor (hBNDF) protein after transplantation of the hBDNF-modified rMSCs (hBDNF-rMSCs) to the adult rats with spinal cord injury (SCI) and discuss the effect of hBDNF-rMSCs on the apoptosis of rat neural cells. Methods A total of 240 adult male Sprague-Dawley rats were randomly divided into sham operation group,SCI group,hBDNF-rMSCs transplantation group and empty vector-rMSCs transplantation group,with60 rats in each group.The SCI model was established by using the modified Allen technique.At day 7 after modeling,an equal volume of hBDNFrMSCs suspension,empty vector-rMSCs suspension and phosphate buffered saline (PBS) were injected through the L4,5 subarachnoid space into the hBDNF-rMSCs group,empty vector-rMSCs group and SCI group,respectively.Then,the injured spinal cord tissues were obtained from each group at days 1,2,3,7 and 14 after transplantation to observe the viability and distribution of rMSCs with enhanced green fluorescent protein gene by fluorescent microscope,measure the expression of hBDNF protein by Western blot and detect the apoptosis of neural cells by TdT-mediated dUTP nick end labeling (TUNEL). Results Both hBDNF-rMSCs and empty vector-rMSCs groups showed green fluorescence expression of rMSCs.The hBDNF protein expression was observed in hBDNF-rMSCs group and changed with time,ie,the expression was detected at day 2 after transplantation,reached the highest level at day 7 and then decreased gradually.Among the hBDNF-rMSCs,empty vector-rMSCs and SCI groups,the number of TUNEL positive cells was the least in hBDNF-rMSCs group,followed by the empty vector-rMSCs group and the number was relatively more in SCI group at days 2,3,7 and 14 after transplantation,with significant differences among groups (P ＜ 0.05). Conclusions Transplantation of the hBDNF-modified rMSCs through subarachnoid approach are able to survive and assemble at the injured spinal cord area and express hBDNF protein.The hBDNF-modified rMSCs can inhibit the apoptosis of neural cells after SCI.

Objective To investigate the effect of FK506 on the expression of axon guidance cue slit-2 after spinal cord injury(SCI)in rats.Methods A total of 75 adult Sprague-Dawley rats were randomly divided into three groups,ie,sham operation group,SCI group and FK506 treatment group.The SCI model was made by using the modified Allen's technique.Then,the rats were sacrificed and the spinal cord was removed at different time points(at days 1,3,7,14 and 28)for detection of the expression of slit-2 by means of reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry.Results The expression of slit-2 changed with time.The expression of slit-2 could be detected at day 1 after SCI,reached the highest at day 7 and then decreased gradually,with higher expression level in the injury group compared with treatment group(P ＜ 0.05).Conclusion Following spinal cord injury,administration of FK506 can up-regulate the expression of slit-2 and may exert important effect on the guidance of the axon regeneration.

Objective To observe the effect of FTY720 on the expressions of Bel-2 and Bax protein and explore the neuroproteetive effect of FTY720 on neuron after acute spinal cord injury in the rats.Methods A total of 75 SD rats were divided randomly into three groups,ie,the test group(treated with FTY720),the injury group(treated with normal saline)and the control group.The test and injury groups were impacted to create T10 spinal cord injury(SCI)by Allen's method.Both the mRNA and protein expressions of Bcl-2 and Bax in the injured spinal cord section were studied respectively with hematoxylin and eosin(HE)staining,immunohistochemical examination and reverse transcription polymerase chain reaction(RT-PCR)technique at different time points(at6 h,12 h,24 h,3 d and 7 d). Results The expressions of Bcl-2 and Bax in the test group and the injury group were higher than that in the control group at all time points.Meanwhile,at 12 h,24 h,3 d and 7 d,the mRNA and protein expressions of Bcl-2 in the test group were significantly higher than that in the injury group(P<0.05),while the mRNA and protein expressions of Bax in the test group were obviously lower than that in the injury group (P<0.05). Condusion Early administration of FTY720(0.5 mg/kg)after spinal cord injmy Canincrease the mRNA and protein expressions of Bcl-2,decrease the expression of Bax and inhibit neuron apoptosis in the injured spinal cord.

Objective To discuss the efficacy and complications of using AngioJet rheolytic thrombectomy in treating acute lower extremity deep vein thrombosis (DVT).Methods The clinical data of 22 patients with acute lower extremity DVT,who were treated with AngioJet rheolytic thrombectomy during the period from February 2015 to August 2016,were retrospectively analyzed.The improvement of clinical symptoms and the thrombus clearance rate were calculated to evaluate the curative effect.The procedure-related complications were documented.Results The clinical symptoms were relieved immediately after operation in all 22 patients.The thigh circumference difference between the affected side and the healthy side decreased from preoperative (4.5±0.6) cm to postoperative (1.0±0.4) cm,the difference in change was statistically significant (P＜0.05).The mean used dose of urokinase was (0.18±0.03) million unit and the average duration of thrombolysis was (4.2±0.7) hours.Complete removal of DVT (＞90％) was achieved in 19 patients,most removal of DVT (50％-90％) in 2 patients,and partial removal of DVT (＜50％) in one patient.After treatment,6 patients developed transient hemoglobinuria,which was relieved after hydration with fluid infusion on the same day.No serious complications such as pulmonary embolism or hemorrhage occurred.Conclusion For the treatment of acute lower extremity DVT,AngioJet rheolytic thrombectomy is safe and effective with less complications.

Objective To transfect PCDNA3.0-HIF1-? into the endothelial progenitor cells(EPCs),in order to provide the foundation for exploring myocardial vascular repairing capacity and angiogenesis of the EPCs transfected with HIF gene.Methods The EPCs were separated by the method of density gradient separation in the human bone marrow,and then the cells were inoculated in the fibronectin-packaged petri dishes complete medium with the concentration of 1?106?mL-1,and cultivated for 14 d. The expressions of the endothelial cell-specific components ecNOS and flk-1 were detected with RT-PCR; the cells were stained with acLDL-Dil and FITC-UEA-1; the PCDNA3.0-HIF1-? plasmid was transfected into the EPCs mediated by LipofectamineTM 2000,the transcription of HIF1-? was determined with RT-PCR; the expression of VEGF was detected with enzyme-linked immunosorbent assay (ELISA); the expression of HIF1-? was detected with Western blotting.Results After cultivated for 5 d,some round monocytes transformated into spindled inherent EPCs;7 d later,there were some cell colonies containing dozens of cells;10 d later,there were obvious cell clones; RT-PCR showed ecNOS band in 548 bp,flk-1 band in 819 bp; acLDL-Dil and FITC-UEA-1 double-staining was positive; RT-PCR showed specific band in 383 bp; ELISA showed the content of VEGF in the supernatant of EPCs transfected with HIF1-? was (20.53?2.33) ?g?L-1,and it was (3.96?1.67) ?g?L-1 in the EPCs transfected without HIF1-?,there was significant difference between two groups (P

Objective To discuss the feasibility and effect of labeled antibody on targeting therapy of systemic lupus erythematosus(SLE) associated thrombocytopenia.Methods MTX-IVIG conjugate was prepared by indirect and direct cross linking,and the binding ability of it to Fc fragment was detected by indirect immunofluor-escence.The killing activity of the conjugate was detected by MTT method using murine macrophage with Fc receptor and strain U937 of human monocytic leukemia cell as targets.Results Conjugation showed stronger cytotoxicity upon target cells than free MTX,and it showed less cytotoxic effect on Fc receptor negative cells compared with the positive ones.IVIG-HSA-MTX restrained the phagocyte of macrophage.The killing effect of IVIG-HSA-MTX was significantly stronger than that of IVIG-MTX.Conclusion The conjugation can show a highly specific cytotoxicity upon mononuclear-macrophage in vitro.