AIM: To construct recombinant adenoviral vector carrying the LEDGFp52 gene by homologous recombination in bacteria and to detect its expression in vitro.METHODS: The LEDGFp52 gene was cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then, the resultant pAdTrack-CMV-LEDGFp52 was cotransfected into BJ5 183 bacteria with the adenoviral backbone plasmid pAdeasy-1. The adenoviral plasmid carrying LEDGFp52 was generated with homologous recombination in bacteria, and the adenoviruses were produced in 293 cells. These 293 cells were then infected with adenoviruses, and the expression of LEDGFp52 was detected by CPE (cytopathic effect) and western blot.RESULTS: The titer of Ad-LEDGFp52 adenoviruses was up to 5×1012 pfu/L after proliferation in 293 cells. LEDGFp52 was expressed efficiently in 293 cells after infection.CONCLUSION: The recombinant adenoviruses vector expressing LEDGFp52 was constructed successfully and can be used in further gene transfection experiments.

AIM: To construct and identify LEDGFp52 eukaryotic expression vector for RNA interference.METHODS: Recombinants were designed and established by targeting gene LEDGFp52 and plasmid pGensil-1 based on LEDGFp52 cDNA sequences of Genomes. Two pairs of oligonucleotides were synthesized according to the Tuschl principle and inserted into plasmid pGenSil-I to generate siRNA eukaryotic expression vector. DH5α strains were transformed, plasmids were extracted, and recombinant vectors were identified by the restriction map and the sequence analysis. The cultured cells were transfected by the recombinant plasmid (pGensil-1-RNA. LEDGFp52-1). At 48 hours after transfection, the whole cell protein was extracted, and the protein level was detected using Western blotting with mouse anti-human LEDGFp52 monoclonal antibody.RESULTS: Recombinant plasmids completely concord with the designs by the restriction map and the sequence analysis,the proteinlevel of LEDGFp52 was down regulated at 48hours after transfecting pGensil-1- LEDGFp52-1 expression vector into HeLa cells, the recombinant eukaryotic expression vectors were successfully constructed.CONCLUSION: siRNA recombinant can be successfully constructed by RNAi technique to inhibit the expression of LEDGFp52.

AIM: To construct the prokaryotic expression vector of rhLEDGFp52 gene,to obtain the rhLEDGF p52 protein.METHODS: rhLEDGFp52 gene was constructed into a prokaryotic expression vector pET30a (+) by recombinant DNA techniques and was identified by enzymatic digestion and sequence analysis. rhLEDGFp52 protein was induced expression by IPTG in E.coli BL21 (DE3), it was tested by Western blot and was purified by Ni-NTA His. Bind. Resin.RESULTS: We Successfully constructed the prokaryotic expression vector of rhLEDGFp52 gene and obtained its expression in E.coli BL21 (DE3),it was expressed in a soluble form and detected up to 34.63% of the total bacterial protein expressed in E.coli BL21 (DE3).Western blot analysis demonstrated that rhLEDGFp52 protein could spicifically integrate with LEDGF-ab. After purified by Ni-NTA His. Bind. Resin, the ultimate concentration of purified rhLEDGFp52 protein was 520μ g/ml and its purity was 87.93%.CONCLUSIONS: rhLEDGF p52 protein was obtained that provides an experimental basis for the further study of the biological function of rhLEDGFp52 protein.

At the time of phage’s discovery, phage therapy was regarded as a possible treatment method against bacterial infection. Although phage therapy was used to treat and prevent bacterial infection in the former Soviet Union and Eastern Europe, it was abandoned by the West in the 1940s with the arrival of the antibiotic era. However, the ongoing evolution of bacterial multidrug-resistance has recently motivated the Western scientific community to reevaluate phage therapy for bacterial infections that are incurable by conventional chemotherapy. With the indepth study of phages, it’s increasingly acknowledged that phages, as the medicine to cure bacterial infection, are convenient, safe and efficient therapeutics. This paper summarizes the recent years’ advanced researches in this area.

OBJECTIVETo establish a method to produce virus-like particles (VLP) of poliovirus type I in Saccharomy cescerevisiae to develop potential novel recombinant vaccine against poliovirus type 1.

METHODSThe genes of P1 and 3CD of poliovirus type I were optimized, synthesized and inserted into expression vector, which was further transfected into Saccharomy cescerevisiae. The extracts of yeast cells were purified by CsCl density gradient centrifugation after induction and cell lysis.

RESULTSElectrophoresis and sequencing analyses showed that the genes P1 and 3CD of poliovirus type I were successfully inserted into expression vector and encode a protein whose amino acid sequences were identical with wide-type genes of poliovirus type I. Electronic microscopy analysis showed that the VLPs of poliovirus type I could be efficiently formed in Saccharomy cescerevisiae.

CONCLUSIONThe VLPs of poliovirus type I could be efficiently produced by co-expression of P1 and 3CD genes in Saccharomy cescerevisiae.

Female ; Gene Expression ; Humans ; Male ; Poliomyelitis ; prevention & control ; virology ; Poliovirus ; genetics ; metabolism ; Poliovirus Vaccines ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism ; Virion ; genetics ; metabolism

One of the important approaches for further prolonging remission duration and eradicating minimal residual disease in acute leukemia is immunotherapy. Four kinds of immunotherapy for acute leukemia are under investigation: (1) monoclonal antibodies, among them, Mylotarg (cytotoxic antibiotic calicheamicin linked to CD33 Mab) is given for the treatment of refractory or relapsed acute myeloid leukemia and molecular relapse in acute promyelocytic leukemia with good results, Campath-1H (antiCD52 Mab) is administered in the treatment of prolymphocytic leukemia and Rituximab (anti-CD20 Mab) in B-PLL with high complete remission rates. Other Mabs under preclinical and clinical trials include anti-IL-2 receptor Mab for the treatment of acute T lymphocytic leukemia, anti-220 kD Mab-6G7 for acute leukemias, recombinant immune toxin BL22 (anti-CD22) for hairy cell leukemia and Mabs labeled with radio-isotopes for different types of acute leukemias; (2) adoptive cellular immunotherapy using cytokine-induced killer cell, alloreactive NK cells, allogeneic or autologous leukemic-specific CD8(+) cytotoxic T lymphocytes, and other immune effector cells; (3) cytokines and other immune modulators comprising IL-2, IL-12, GM-CSF, CD40L, FLT-3L and thalidomide and its derivatives; (4) leukemia vaccines of several different formulations including antigen-specific, leukemia cell-based, leukemia antigen-pulsed dendritic cell (DC) and leukemia-derived DC vaccines, the latter two formulations are more attractive. In conclusion, up to now, the most effective example of immunotherapy in acute leukemia is provided by the administration of Mabs, and the majority of other approaches in immunotherapy for acute leukemia although promising, need further studies.
Acute Disease ; Adoptive Transfer ; methods ; Antibodies, Monoclonal ; immunology ; therapeutic use ; Cancer Vaccines ; therapeutic use ; Humans ; Immunotherapy ; methods ; trends ; Leukemia ; immunology ; therapy

Gout ; therapy ; Humans ; Integrative Medicine

Gastrointestinal neuroendocrine tumor(GI-NET)originates from peptide neurons and neu-roendocrine cells in gastrointestinal tract,and secrets peptide hormones,leading to carcinoid syndrome which rarely happens in clinical practice. Because of the improvement of diagnostic method and understanding of this rare disease,the morbidity is rising in recent years. The main treatments of GI-NET are surgery and compre-hensive therapy,consisting of chemotherapy and targeted therapy.

Objective To investigate the clinical value of a three-dimensional motion analysis system by using the body segmental method for testing the balance of hemiplegic stroke patients.Methods Twenty stroke patients with hemiplegia were measured using the lower extremity Fugl-Meyer motor assessment (FMA-L),the Brunel balance assessment (BBA),the Berg balance sale (BBS) and a 5 m timed up-and-go test (5m-TUGT).The three-dimensional motion analysis system using the body segmental method was applied in three positions-sitting,standing and walking.Spearman's rank correlation coefficient was used to determine the extent of correlation between the values measured by the three-dimensional motion analysis system and the FMA-L,BBA,BBS and 5m-TUGT results.Results Some motion analysis variables (MSAx,MSVx,MSAy,MSVy and SPxy,SP3-D) are recorded in sitting and standing,while the center of gravity (COG) swing in the horizontal plane and some other variables (MSAx,MSVx and SPxy and SP3-D) are measured while walking.Anterior-posterior COG swing had a high negative correlation with the FMA-L,BBA and BBS scores and a high positive correlations with 5m-TUGT times.But except in sitting,MSAz and MSVz were both uncorrelated with FMA-L,BBA or BBS scores or with 5m-TUGT times.COG swing in the horizontal plane in sitting and standing apparently correlates with lower extremity motor function,balance,and walking ability.However,only the X axis swing parameters of the COG while walking correlated with lower extremity motor function,balance or walking ability.Conclusions A three-dimensional motion analysis system using the body segmental method can be used clinically to monitor patients' balance in real time and dynamically in different positions and activities,and it can be used to predict motor function and balance control in hemiplegic stroke patients.

AIM: To dynamically observe the feeling change of the photorecrptor layer in the eyes with acute central serous chorioretinopathy ( CSCR ) krypton laser treatment by fourier - domain optical coherence tomography ( FD - OCT), and to study their correlation with the chang of vision.
METHODS: This is a retrospective case series study. The clinical diagnosis of 52 patients with monocular initial onset of central serous chorioretinopathy, krypton laser photocoagulation before treatment, after 1, 2, 4, 6, 8wk, 6mo, FD - OCT were performed to observe the morphological changes characteristic of photoreceptor layer and changes in vision.
RESULTS: After 1wk treatment, all cases were improved; 2wk, 6 cases were cured; 4wk, 38 cases were cured; 6wk, 41 cases were cured; 8wk, 45 cases were cured, the OCT showed macular retinal neuroepithelial layer ( RNL ) from fully absorbed; 6mo with the same 8wk. Before and after treatment in patients with best corrected visual acuity and from the height difference between the macular region of RNL was statistically significant (P<0. 05), there was a correlation between the changes of visual acuity after treatment and the macular detachment of RNL height (P<0. 05), Photoreceptor layer of complete and incomplete best corrected visual acuity difference was statistically significant (P<0. 01).
CONCLUSION: FD-OCT can dynamicaly observed acute central serous chorioretinopathy krypton laser treatment of photoreceptor ultrastruture changes. Photoreceptor layer of complete and incomplete best corrected visual acuity difference was statistically significant (P<0. 01).