Objective:To investigate influences of two different HLA-B antigens expressed on K562 cells on receptors expression of NK cells from peripheral blood lymphocytes.Methods:Studied the alteration of the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells before and after PBMC interaction with K562 cells for 24 hours,and also compared the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells after PBMC interaction with two different kind of K562 cells transfected with HLA-B39 and HLA-B51 respectively.Results:After PBMCs were incubated with K562 cells for 24 hours,the percentage of CD16+CD56+ cells and the percentage of KIR3DL1+ cells were both increased.However,after PBMCs were incubated with K562-HLA-B51 cells for 24 hours,the percentage of KIR3DL1+ cells and the percentage of CD16+CD56+ cells were both decreased in comparison with that interaction with K562-HLA-B39 cells.Conclusion:CD16 up-regulation was associated with an up-regulation of inhibitory receptors(KIR3DL1).The interaction between HLA-Bw4 and KIR3DL1 would down-regulate the expression of KIR3DL1.In addition,KIR3DL1 down-regulation was associated with down-regulation of activating receptors(CD16).

Objective To investigate whether MICA gene exon 2,3 and 4 polymorphism is associated with seronegative spondylarthropathies (SpA) or not in Chinese Han population.Methods All 199 B27 positive patients with SpA and 183 randomly ethnically matched healthy controls,and 12 B27 positive controls were enrolled to detect the MICA genotype from its exons 2,3 and 4 by using PCR SSOP method in Shanghai area.Results The MICA007 allele frequency was significantly more in the patient group (18 0%) than in the randomly healthy control group (6 6%) (RR=3 04, P =0 000 045).However,the MICA007 allele frequency was not significantly higher in the B27 positive patient group than in the B27 positive control group.Conclusion The MICA007 allele itself may not be the real disease susceptibility gene involved in the development of ankylosing spondylitis.The increased frequency of MICA007 allele is supposed to be due to a strong linkage disequilibrium between MICA and HLA B genes.

Objective:To analyze the frequencies of HLA-A*0201 restricted CEA-specific CD8+T cells, HLA-A*0201/FLUmp tetramer and HLA-A*0201/CAP-1 tetramer were applied in patients with colorectal cancer.Methods: Lymphocytes from peripheral blood and lymph node,1×106 cells/ml,were incubated with 1μg HLA-A*0201/peptide tetramers and anti-CD8 for 1 h at 25 coseperately.The cells were then washed in PBS.Next,the cells were illuminated by detecting frequencies of FLUmp-specific CD8+T cells and CAP-1-specific CD8+T cells with flow cytometry.Results: HLA-A*0201/peptide were used to detect CAP-1 or FLUmp-specific CD8+T cells,which were analyzed either healthy individuals or patients with colorectal cancer.We did not find differences in average frequencies of FLUmp-specific CD8+T cells between 11 HLA-A*0201+patients with colorectal cancer and 14 HLA-A*0201+healthy individuals [ ( 0.671 ±0.421 )%, ( 0.564 ±0.408 )%].But the frequencies of CAP-1-specific CD8+T cells of HLA-A*0201+patients with colorectal cancer showed higher than HLA-A*0201+healthy individuals [ ( 2.409 ± 2.385 )%, ( 0.020 ± 0.021)%respectively],which was statistically significant(P=0.008).Conclusion:The frequencies of CAP-1-specific CD8+T cells in PBMC from peripheral blood and lymph node of HLA-A*0201+patients were increased,showed CEA-specific CTs has a vital role in colorectal cancer.

Objective To summarize and analyze the results of our university graduates' participation in the national medical licensing examination in order to investigate the clinical medical teaching reform.Methods There were 770 candidates who took the NMLE from 2011 to 2013.Excel 2007 was used to statistically analyze the total score,passing rate,average score and mastery rate for each subject,and average score and mastery rate at different cognitive level.Results In these three years,our candidates' total pass rate was higher than the national average total pass rate,respectively higher 17.11％,18.10％,17.82％;the pass rate of practical skills examination was also higher than the national average pass rate,respectively higher 7.30％ and 7.19％ and 5.90％;the pass rate of theory examination was higher than the national average pass rate,respectively higher 12.91％,12.97％ and 14.29％.The average score and mastery rate for each subject,the average score and mastery rate at different cognitive level were also all higher than those of the national average of medical colleges.Conclusions National medical licensure examination can be used as an important reference,through optimization of medical curriculum and teaching mode,optimization of clinical practice skills training system and optimization of clinical practical skill evaluation system etc.,to further improve the clinical medical personnel training mode.

Objective To investigate whether the killer cell immunoglobulin like receptor (KIR) gene polymorphisms are associated with seronegative spondylarthropathies.Methods All 197 HLA B27 positive patients with seronegative spondyloarthro pathies (SpA) and 83 randomly ethnically matched healthy controls were enrolled to detect the KIR genotype using PCR SSP in Shanghai area.Results The KIR3DL1 gene frequency was significantly lower in the patient group (0 763) than in the control group (1 00) (RR=0 76, P =0 003).Meanwhile,the gene frequencies of two pseudogenes (KIR2DP1,KIR3DP1) were significantly higher in SpA group than in healthy control group (RR=1 1, P =0 004).Conclusion There may be a negative association between pathogenesis of SpA and KIR3DL1 gene.The function of KIR3DL1 molecule should be investigated deeply.

AIM: To study the effects of recombinant soluble MHC class Ⅰ chain-related protein A (sMICA) on the cytotoxicity, secretion of IFN-γ, proliferation and apoptosis of peripheral NK cells. METHODS: After NK cells were cocultured with recombinant soluble MICA proteins overnight, the cytotoxicity of NK cell on target cells was detected by flow cytometry. The supemant was collected to determine the concentration of IFN-γ by ELISA. The proliferation of NK cells to sMICA was detected by MTS/PMS. NK cells were labeled with annexin V and PI to analyze their apoptosis. RESULTS: Soluble MICA inhibited the cytotoxicity of NK cells and down-regulated the secretion of IFN-γ, but it showed no effects on the proliferation and apoptosis of freshly isolated peripheral NK cells. CONCLUSION: The soluble MICA shedding from tumor cells could be a pathway of cancer immune evasion by down-regulating the biologic activities of NK cells.

AIM: To detect the effect of conjunction matrigel with mammary fad pat(MFP)implantation on the tumorigenesis, proliferation, apoptosis and metastasis of Her2 positive and negative breast cancer model. METHODS: The Her2 positive BT 474 and Her2 negative MDA-MB 231 breast cancer cells were injected into MFP of nude mice with or without matrigel to establish breast cancer model. The tumor volume was measured every 3 d. Followed up for 30 d after implantation, the nude mice were killed and the tumors and associated organs were dissected for pathological sectioning and staining with hematoxylin and eosin. The time of tumor formation and the tumorigenesis were determined after implantation. The tumor volume and metastasis rate were calculated and compared with each other. The proliferation and apoptosis of Her2 positive and negative tumors were also determined. RESULTS: Matrigel and MFP implantation technology shortened the time of tumorigenesis significantly(P<0.01). The tumorigenesis rate of BT 474 and MDA-MB 231 breast cancer cells did not show any different(P>0.05). The metastasis rate of MDA-MB 231 breast cancer cells were improved from 25.0% to 37.5%(P<0.05). CONCLUSION: Matrigel and MFP implantation can be combined to shorten the time of tumor formation by two kinds of breast cancer cells, and improve the metastasis of Her2 negative MDA-MB 231 cells. Using matrigel does not show any effect of proliferation and apoptosis on Her2 positive and negative breast cancer cells.

Angiopoietin-like protein (ANGPTL) 4 is involved in the regulation of multiple tumor growth and metabolism pathways and plays a multifunctional role in many aspects of tumorigenesis such as growth,metastasis,apoptosis,angiogenesis and tumor microenvironment.As one of the members of ANGPTL family,it is confirmed that ANGPTL4 both has a tumor-promoting and tumor-inhibiting effect on digestive system neoplasms in recent years.This dual role may be related to its involvement in regulating different tumor signaling pathways.ANGPTL4 is expected to be a new target for diagnosis and treatment of digestive system neoplasms.

BACKGROUND/AIMS: The functional variant (rs56109847) in the 3'-untranslated regions (3'-UTR) of the serotonin receptor 3E (HTR3E) gene is associated with female diarrhea predominant irritable bowel syndrome (IBS-D) in British populations. However, the relationship of the polymorphism both to HTR3E expression in the intestine and to the occurrence of Chinese functional gastrointestinal disorders has yet to be examined. METHODS: Polymerase chain reaction amplification and restriction fragment length polymorphism analyses were employed to detect polymorphisms among Chinese Han women, particularly 107 patients with IBS-D, 99 patients with functional dyspepsia (FD), 115 patients with mixed IBS and 69 patients with IBS-D + FD. We also assessed microRNA-510 (miR-510) and HTR3E expression in human colonic mucosal tissues with immunohistochemistry and other methods. Dual-luciferase reporter assays were conducted to examine the binding ability of miR-510 and HTR3E 3'-UTR. RESULTS: Genotyping data showed the variant rs56109847 was significantly associated with IBS-D, but not with FD, mixed-IBS, or FD + IBS-D. HTR3E was abundantly expressed around the colonic mucosal glands but less expressed in the stroma. miR-510 expression decreased, whereas HTR3E expression increased in the colonic mucosal tissue of patients with IBS-D compared with those in controls. HTR3E expression was significantly higher in patients with the GA genotype than that in patients with the GG genotype. The single-nucleotide polymorphisms disrupted the binding site of miR-510 and significantly upregulated luciferase expression in HEK293 and HT-29 cells. CONCLUSIONS: The single-nucleotide polymorphisms rs56109847 led to reduced microRNA binding and overexpression of the target gene in intestinal cells, thereby increasing IBS-D risk in the Chinese Han population. The decreased expression of miR-510 might contribute to IBS-D.
Asian Continental Ancestry Group* ; Binding Sites ; Colon ; Diarrhea* ; Dyspepsia ; Female ; Gastrointestinal Diseases ; Genotype ; HT29 Cells ; Humans ; Immunohistochemistry ; Intestines ; Irritable Bowel Syndrome* ; Luciferases ; MicroRNAs ; Mucous Membrane ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Serotonin*

OBJECTIVETo detect genetic polymorphism in the exons 2 to 4 of the MICA gene in Chinese Han population in Shanghai, Dai population in Yunnan province and Uygur population in Xinjiang province respectively.

METHODSDNA samples from 183 random healthy individuals in Han population, 41 in Dai population and 66 in Uygur population were genotyped by using the polymerase chain reaction (PCR) and sequence-specific oligonucleotide probing (SSOP) method.

RESULTSIn total, 10, 7 and 9 alleles of MICA were observed in Han, Dai and Uygur population respectively. MICA*008 was the most common allele in the population of both Han and Uygur, whereas MICA*010 was the most popular one in Dai population. Han and Dai had a bit similar distribution pattern (Chi-square=12.809 P=0.046), in contrast with Han to Uygur (Chi-square=58.499 P=0) and Dai to Uygur (Chi-square=49.273 P=0).

CONCLUSIONMICA gene displayed different allele distributions in different populations.

Alleles ; China ; Exons ; genetics ; Gene Frequency ; Genotype ; Geography ; Haplotypes ; Histocompatibility Antigens Class I ; genetics ; Humans ; Linkage Disequilibrium ; Polymorphism, Genetic