Animals ; Epitopes, T-Lymphocyte ; immunology ; Equine Infectious Anemia ; immunology ; virology ; Horses ; Infectious Anemia Virus, Equine ; immunology ; physiology ; T-Lymphocytes, Cytotoxic ; immunology ; physiology

NAC transcription factors involved in plant growth and development, as well as responses to biotic and abiotic stress. RNAi Vectors for SmNAC transcription factors of Salvia miltiorrhiza was constructed by using Gateway cloning technology, in order to further study the function of SmNAC1 transcription factor. According to Gateway cloning technology, the specific fragments of SmNAC1 containing attB adapter was amplified by PCR using ultra-fideling phusion polymerase of NEB. By the BP recombination reaction, the PCR product containing attB was transferred to an donor vector (pENTR/SD/D-TOPO). Finally, SmNACi specific gene was cloned into pK7GWIWG2D plant expression vectors by LR recombination reaction. Experimental results showed that Gateway cloning technology provide a rapid and highly efficient way to clone the interested gene.
Cloning, Molecular ; methods ; Genetic Vectors ; genetics ; Plant Proteins ; genetics ; Polymerase Chain Reaction ; RNA Interference ; Reproducibility of Results ; Salvia miltiorrhiza ; genetics ; Transcription Factors ; genetics

Carrier Proteins ; metabolism ; physiology ; HIV-1 ; physiology ; Models, Biological ; Proteins ; metabolism ; physiology ; Retroviridae Infections ; virology

Base Pairing ; Dimerization ; Genome, Viral ; RNA, Viral ; chemistry ; genetics ; Retroviridae ; chemistry ; genetics

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, a full-length cDNA sequence of SQS2 from S. miltiorrhiza f. alba was cloned by the method of reverse transcription polymerase chain reaction (RT-PCR). The SmSQS2 cDNA sequence was obtained, this sequence is named SmSQS2 and its GenBank registration number is KM244731. The full length of SmSQS2 cDNA was 1245 bp, encoding 414 amino acids including 5'UTR 115 bp and 3'UTR 237 bp. Sequence alignment and phylogenetic analysis demonstrated that SmSQS2 had relative close relationship to the SQS2 of S. miltiorrhiza. The induction of E. coli [pET28-SQS2] in different temperature, induction time, IPTG concentrations and density of inducing host bacterium (A600) were performed, Shaking the culture at 30 degrees C until the A600 is approximately 0.6 and add IPTG to final concentration of 0.2 mmol x L(-1), and then the optimal expression of SmSQS2 recombinant protein were accumulated after the induction time of 20 h. The research provided important base for the study of sterol and terpene biosynthesis of SQS2 in S. miltiorrhiza f. alba.
Cloning, Molecular ; Farnesyl-Diphosphate Farnesyltransferase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; classification ; enzymology ; genetics ; Sequence Alignment

AIM:To investigate the effects of silymarin on lipopolysaccharide(LPS)-induced acute lung injury in rats and its possible molecular mechanisms.METHODS: Fifty-eight male SD rats,weighting 230-250 g,were divided into four groups randomly: normal control(n=12);acute lung injury group(n=15),receiving intravenous LPS(O55∶B5,5 mg/kg);silymarin alone group(50 mg/kg,n=15);intervention group(n=16,receiving silymarin 50 mg/kg and LPS 5 mg/kg).The specimens were collected 6 hours later.The following changes,including blood gas analysis,the lung wet/dry weight ratio,the pulmonary vascular permeability,histological manifestations,lung tissue myeloperoxidase activity,the levels of TNF-?,IL-1?,MCP-1 and SOD,GSH-Px as well as malonaldehyde and conjugated diene in plasma and lung tissue,were observed.RESULTS: Compared with control group,the lungs of the rats in LPS treatment group showed significant hyperemia and spotted hemorrhage.The inflammatory granulocyte infiltrating,diffused alveolar septum thickening and spotted hemorrhage were observed in pathological examinations.The lung wet/dry weight ratio and Evans blue content(per gram) increased significantly after LPS treatment.The myeloperoxidase activity in plasma and lung tissue,the levels of TNF-?,IL-1?,MCP-1 and SOD,GSH-Px as well as malonaldehyde and conjugated diene were increased significantly in LPS treatment group.However,in intervention groups,all the above-mentioned measurements were reversed significantly by silymarin treatment compared with LPS treatment group.CONCLUSION: Silymarin may decrease inflammatory reaction and oxidative stress,and further decrease lung damage induced by LPS in rats,all indicating protection of silymarin against acute lung injury.

Objective: To investigate the effects of grasp seed procyanidins(GSP,原青花素) on lipopolysaccharide(LPS)induced acute lung injury(ALI) in rats with renal function damage and the related possible molecular mechanisms.Methods: The homogenates of lung and kidney were prepared and venous blood were collected at 6 hours after injection of LPS and medicine.The changes of contents of creatinine(Cr),blood urea nitrogen(BUN),lactic acid(Lac) and nitric oxide(NO) in the blood were measured.The enzyme linked immunosorbent assay(ELISA) was used to measure the levels of tumor necrosis factor?(TNF-?),interleukin-1?(IL-1?),monocyte chemoattractant protein-1(MCP-1) and IL-6 in the serum,lung and renal cortex tissue homogenate in various groups.The histopathological changes of lung tissues were observed.The pulmonary vascular permeability and the lung wet/dry(W/D) weight ratio were determined;the malonaldehyde(MDA) content,Na+K+-ATPase,superoxide dismutase(SOD),myeloperoxidase(MPO) and glutathion peroxidase(GSH-Px) activities in lung and renal tissues were also determined.Changes of mitogen activated protein kinase(MAPKs) were detected by Western blotting,and the combination activity of nuclear factor-?B(NF-?B) to DNA was detected by electrophoretic mobility shift assay (EMSA) in lung tissues.Results: ①Compared with the normal rats in control group,the lungs of the rats in LPS treatment group and GSP group had significant hyperemia and spotted hemorrhage.The inflammatory granulocyte infiltration,diffuse alveolar septum thickening and spotted hemorrhage were observed in the pathological examinations,while in LPS plus GSP group the above mentioned pathological changes were milder.②Compared with control group,the lung W/D and pulmonary vascular permeability were much higher in the LPS treatment groups(P

Immunomodulatory drug lenalidomide is a synthetic compound derived by modifying the chemical structure of thalidomide to improve its potency and reduce its side effects. Lenalidomide is a 4-amino-glutamyl analogue of thalidomide that has emerged as a drug with activity against various hematological and solid malignancies. It is approved by FDA in USA for clinical use in myelodysplastic syndromes with deletion of chromosome 5q and multiple myeloma. Studies have shown that lenalidomide exert anti-tumor activity probably by various mechanisms in hematologic malignancies, such as immunomodulation, anti-angiogenesis and effects on signal transduction. In this article, the progresses of study on these problems are reviewed.
Hematologic Neoplasms ; immunology ; Humans ; Immunologic Factors ; Thalidomide ; analogs & derivatives ; pharmacology

AIMTo study the chemical constituents in the mycelia of Hypomyces sp..

METHODSSilica gel column chromatography was employed for the isolation and purification. Chemical and spectral methods were used to determine the structures of the isolated compounds.

RESULTSTwo compounds were isolated and identified as: hypomycin C (I) and hypomycin D (II).

CONCLUSIONCompounds I and II are new compounds.

Chromatography, Gel ; methods ; Fermentation ; Hypocreales ; chemistry ; Molecular Conformation ; Molecular Structure ; Mycelium ; chemistry ; Perylene ; analogs & derivatives ; chemistry ; isolation & purification ; Quinones ; chemistry ; isolation & purification

WRKY transcription factor is one of the Zinc finger proteins which contains a highly conserved WRKY domain and is a family of the plant-specific transcription factor. The plasmid pET28a-SmWRKY1 harboring Salvia miltiorrhiza WRKY1 (SmWRKY1) gene was successfully transformed and expressed in Escherichia coli BL21 (DE3). The conditions on protein expression of SmWRKY1 in E. coli, including induction duration, temperature, IPTG concentration and the E. coli concentration were optimized. The results showed that the highest protein expression of SmWRKY1 was obtained at 24 hours after the E. coli was cultured in the presence of 0.2 mol x L(-1) IPTG at 20 degrees C with A600 values of 1.0-1.5. This recombinant histidine-tagged protein was expressed at 2.454 g x L(-1) as inclusion body, which was first extracted using urea, and then purified by Ni2+ affinity chromatography and identified by SDS-PAGE. The expression of SmWRKY1 in E. coli was further confirmed by western blotting analysis.
Blotting, Western ; Cloning, Molecular ; DNA-Binding Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; chemistry ; genetics ; metabolism ; Molecular Weight ; Plant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Salvia miltiorrhiza ; genetics