Objective To analyze the nucleotide sequences of novel HLA class I le,B*1316.Methods Routine sequence-specific oligonucleotide(SSO) typing and sequencing based typing(SBT) was used.Results The B*1316 allele differs from B*1302 by one nucleotide substitution in exon 3: T to A at nt position 184,which results in an amino acid substitution at codon 62 from Val to Glu.Conclusion A novel HLA class I allele,B*1316 has been identified,and was officially recognized by WHO Nomenclature Committee in April 2006.

AIM: To examine expression of macrophage migration inhibitroy factor (MIF) gene and protein in macrophages induced by oxidized low density lipoprotein (ox-LDL). METHODS: Macrophages were incubated with ox-LDL at the concentration of 150 mg/L for time course (0-36 h) and with ox-LDL at the different concentrations (0-300 mg/L) for 24 h, expression of MIF mRNA and protein were detected by RT-PCR and ELISA. RESULTS: The results showed that ox-LDL increased MIF gene and protein expression in macrophages in a dose and time-dependent manner. After the exposure of macrophage to ox-LDL, the expression of MIF mRNA level increased consistently with protein. CONCLUSION: MIF may play an important role in atherosclerosis. [

Objective] To determine the nucleotide sequence of the 3′\|termal of the RESA gene Plasmodium falciparum isolate FCC1/HN, and find out the differences of the sequences of RESA gene among isolate FCC1/HN,FC27,NF7 and Palo Alto. [Methods] 3′\|terminal fragment of RESA gene of P.falciparum isolate FCC1/HN was amplified by PCR method, then was cloned into pMD18\|T vector. The recombinant was screened and identified by BamHI+XhoI and PCR technique. The nucleotide sequence of the 3′\|terminal of the RESA gene was determined by the dideoxy chain termination method. DNASTAR and BLAST software were used to compare and analyze the RESA gene sequences among the different isolates. [Results] The 3′\|termal fragment of the RESA gene with about 846 bp was specifically amplified by PCR, the recombinant pMD18\|T\|RESA was successfully constructed. Different degrees of diversity of the RESA gene sequences were found among P.falciparum isolates FCC1/HN、FC27,NF7 and Palo Alto. [Conclusion] There were differences in the sequences of RESA gene among the P.falciparum isolate FCC1/HN and three other isolates (FC27,NF7 and Palto alto).

AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress ( ERS) model in CASMCs of SD rats.METHODS:CASMCs were cultured by tissue explant method .The morphological characteristics were observed under optical micro-scope.The marker proteins of CASMCs , including α-SMA and SM-MHC, were identified by immunofluorescence tech-nique.The protein expression levels of BiP and CHOP , the marker molecules of ERS, were determined by Western blot . RESULTS:The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical hill and valleygrowth pattern of CASMCs at 9~10 d.The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed .The results of Western blot showed that the protein expression of BiP and CHOP in TG ( 1 and 2 μmol/L ) treatment groups was increased compared with control group .Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION:CASMCs can be successfully cultured by tissue explant method .ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.

AIM:MicroRNAs ( miRNAs) were recognized to play significant roles in cardiac hypertrophy .But, it remains unknown whether cyclin/Rb pathway is modulated by miRNAs during cardiac hypertrophy .This study investigates the potential roles of microRNA-1 (miR-1) and microRNA-16 (miR-16) in modulating cyclin/Rb pathway during cardiomyocyte hypertrophy .METHODS:An animal model of hypertrophy was established in a rat with abdominal aortic constriction (AAC).In addition, a cell model of hypertrophy was also achieved based on PE-promoted neonatal rat ventricular cardiomyocyte .RESULTS:miR-1 and-16 expression were markedly de-creased in hypertrophic myocardium and hypertrophic cardiomyocytes in rats .Overexpression of miR-1 and -16 suppressed rat cardiac hypertrophy and hypertrophic phenotype of cultured cardiomyocytes .Expression of cyclins D1, D2 and E1, CDK6 and phosphorylated pRb was increased in hypertrophic myocardium and hypertrophic cardiomyocytes , but could be reversed by enforced expression of miR-1 and -16.CDK6 was validated to be modulated post-transcriptionally by miR-1, and cyclins D1, D2 and E1 were further validated to be modulated post-transcriptionally by miR-16.CONCLUSION: Attenuations of miR-1 and -16 provoke cardiomyocyte hypertrophy via derepressing the cyclins D1, D2, E1 and CDK6, and activating cyclin/Rb pathway.

AIM:To investigate the effect of miR-214 on cardiomyocyte hypertrophy and the expression of the potential target genes . METHODS:A cell model of hypertrophy was established based on angiotensin-Ⅱ( Ang-Ⅱ)-induced neonatal mouse ventricular car-diomyocytes (NMVCs).Dual luciferase reporter assay was performed to verify the interaction between miR-214 and the 3’ UTR of MEF2C.The expression of MEF2C and hypertrophy-related genes at mRNA and protein levels was determined by RT-qPCR and Wes-tern blotting, respectively.RESULTS:The expression of ANP, ACTA1,β-MHC and miR-214 was markedly increased in Ang-Ⅱ-in-duced hypertrophic cardiomyocytes .Dual luciferase reporter assay revealed that miR-214 interacted with the 3’ UTR of MEF2C, and miR-214 was verified to inhibit MEF2C expression at the transcriptional level .The protein expression of MEF2C was markedly in-creased in the hypertrophic cardiomyocytes .Moreover, miR-214 mimic, in parallel to MEF2C siRNA, inhibited the expression of hy-pertrophy-related genes in Ang-Ⅱ-induced NMVCs.CONCLUSION:MEF2C is a target gene of miR-214, which mediates the effect of miR-214 on attenuating cardiomyocyte hypertrophy .

The effects of ketamine on transient outward potassium current (I(to)) of isolated human atrial myocytes were investigated to understand the mechanism of part of its effects by whole-cell patch-clamp. Atrial myocytes were enzymatically isolated from specimens of human atrial appendage obtained from patients under going cardiac valve displacing. Ito is recorded in voltage-clamp modes using the patch-clamp technique at room temperature. Currents signals were recorded by an Axopatch 200B amplifier with the Digidata 1322A-pClamp 9.0 data acquisition system. Ketamine decreased I(to) of human atrial myocytes in a dose-dependent manner. The current-voltage curve was significantly lowered, 30, 100, 300, and 1000 micromol x L(-1) ketamine decreased respectively I(to) current density about (13.62 +/- 0.04)%, (38.92 +/- 0.05)%, (72.24 +/- 0.10)% and (83.84 +/- 0.05)% at the potential of 50 mV, with an IC50 of 121 micromol x L(-1). The I(to) activation curve, inactivation curve and the recovery curve were not altered by ketamine. So, ketamine concentration-dependently decreased I(to) of human atrial myocytes.

[Objective] To construct a eukaryotic expression plasmid containing a gene encoding a 41-3 kilodalton blood stage antigen (p41-3) of Plasmodiu falciparum isolate FCC1/HN, and to determine the sequence of p41-3 gene and analyze the homology of the sequences of 341-3 gene of different P. falciparum isolates. [ Methods] Two pairs of primers were designed according to the known sequence of p41-3 gene. Using PCR technique, the p41-3 gene was obtained by amplification from genomic DNA of isolate FCC1/HN. By cloning target gene into a eukaryotic expression vector, pcDNA3, a recombinant plasmid pcDNA3-p41-3 was con structed and trarsferred into E. coli DH5α. The positive clones were screened and identified by agarose gel electrophoresis, endonu clease digestion and PCR technique. The correct recombinant plasmid pcDNA3-p41-3 was used as template, and the nucleotide se quence of p41-3 gene was determined by the dideoxy chain termination method. Using softwares to analyze the structure and sequence homology of p41-3 gene between isolate FCC1/HN and FCBR. [Results] The p41-3 gene was specifically amplified from genomic DNA of Plasmodiumm falciparum isolate FCC1/HN, and the correct recombinant plasmid pcDNA3-p41-3 was screened and identi fied. The result of sequence determination showed that the p41-3 gene of isolate FCC1/HN was 2 137 base pairs in full length, encod ing 375 amino acids. Isolate FCC1/HN and isolate FCBR exhibited 98.98 % homology in the nucleiotide sequences and 99.73 % ho mology in the encoded anino acids of p41-3 gene. [Conclusion] The eukaryotic expression plasmid pcDNA3-p41-3 is successfully con structed and nucleotide sequence of p41-3 gene of isolate FCC1/HN is determined. The p41-3 genes of isolate FCC1/HN and isolate FCBR share quite high homology.

Objective:To observe the killing effect of irradiated peripheral blood mononuclear cells (PBMCs) at low dose combined with apogossypolone (ApoG2) on cultured human prostate cancer PC-3 cells.Methods:Human PBMCs were irradated by gamma ray at 1 gray,the irradiation dose rate was 17 Gy/min.The experiment were divided into PC-3 tumor cell control group,PC-3 cells with irradiated and non-irradiated PBMCs co-culture groups,ApoG2 treatment group,irradiated PBMCs and ApoG2 co-treatment group.Acridine orange/ethidium bromide (AO/EB) staining and MTT method were used to observe the killing effect of PBMCs and/or ApoG2.Results:The killing activity of irradiated PBMCs group and ApoG2 treatment group were obviously increased and were higlaer than that of non-irradiated group (P＜0.05).The killing activity of combined group were much higher than that of irradiated group and ApoG2 treatment group (P ＜0.01 ).Conclusion:Irradiated PBMCs at low dose combined with ApoG2 can enhances the anti-tumor effects markedly.

AIM:To determine circular RNA (circRNA) profiles in the diabetic mouse myocardium , and to investigate the effect of circRNA_000203 on fibrotic phenotypes in cardiac fibroblasts .METHODS:Masson trichrome stai-ning was performed on the myocardium of the diabetic db /db mice and the non diabetic db/m control mice .circRNA ex-pression profile in the diabetic myocardium was detected by circRNAs microarray .The expression of circRNA_000203 was determined by real time fluorescence quantitative PCR ( RT-qPCR ) .Recombinant circRNA_000203 adenovirus was pre-pared for enforced the expression of circRNA_000203 in mouse cardiac fibroblasts.The expression of Col1a2, Col3a1andα-SMA was determined in circRNA_000203-modified cardiac fibroblasts , respectively .RESULTS:Masson trichrome stai-ning showed that fibrosis was increased in the diabetic mouse myocardium .The results of circRNA array detection revealed that circRNAs were dysregulated in the diabetic myocardium .circRNA_000203 was up-regulated in the diabetic myocardi-um.Significant over-expression of circRNA_000203 was achieved in the cardiac fibroblasts after infection with the recombi-nant circRNA_000203 adenovirus.The mRNA and protein expression of Col1a2, Col3a1 and α-SMA was significantly in-creased in the cardiac fibroblasts with over-expression of circRNA_000203.CONCLUSION:circRNA_000203 is up-regu-lated in the diabetic mouse myocardium .It has pro-fibrotic effect on the cardiac fibroblasts .