CTLA4基因多态性与肺结核易感相关性研究进展

探讨CTLA-4基因多态性与肺结核易感相关性,应用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析病例组和对照组中CTLA4基因功能性多态位点（＋49A/G、-1661A/G、-1772T/C）;研究结果显示CTLA-4能下调免疫应答,抑制T淋巴细胞活化;CLTA-4具有基因多态性,影响mRNA转录的稳定性;CTLA-4基因的功能和表达对由T细胞介导的肺结核免疫有重要影响;揭示了结核病易感相关性的分子基础,找到有效控制肺结核的方法。     

Association of Graves＇ disease and Graves＇ ophthalmopathy with the polymorphisms in promoter and exon 1 of cytotoxic T lymphocyte associated antigen-4 gene

Objective： To investigate the association of Graves＇ disease and Graves＇ ophthalmopathy with the C/T transition polymorphism at position -318 of promoter and the A/G transition polymorphism at position 49 of exon 1 within cytotoxic T lymphocyte associated antigen-4 （CTLA-4） gene. Methods： Thirty-three patients with ophthalmopathy of Graves＇ disease, fifty-six Graves＇ patients without ophthalmopathy and sixty normal subjects as control were involved in the present case-control study. The polymorphisms were evaluated by polymerase chain reaction fragment length polymorphism （PCR-RFLP）. Comparisons were made of gene frequencies and allele frequencies between the groups. Results： The gene frequencies of CT and allele frequencies of T were much higher in Graves＇ patients with ophthalmopathy than that in the group without ophthalmopathy （P=-0.020, P=-0.019）. The gene frequencies of GG and allele frequencies of G in patients with Graves＇ disease were significantly increased as compared with control group （P=0.008, P=0.007）. The data suggest that smokers with Graves＇ disease seemed to be more predisposed to ophthalmopathy than non-smokers （P=0.018）. Conclusion： Our results suggest that an allele of T at position -318 of promoter is associated with genetic susceptibility to Graves＇ ophthalmopathy while an allele of G at position 49 of exon 1 is associated with genetic susceptibility to Graves＇ disease instead. Smoking is believed to be a major risk factor for ophthalmopathy.     

Relationships between endothelial nitric oxide synthase gene polymorphisms and osteoporosis in postmenopausal women

Objective: To investigate the relationships between endothelial nitric oxide synthases (eNOS) G894T and 27 bp-variable number tandem repeat (VNTR) gene polymorphisms and osteoporosis in the postmenopausal women of Chinese Han nationality. Methods: In the present study, 281 postmenopausal women from Xi'an urban area in West China were recruited, and divided into osteoporosis, osteopenia, and normal groups according to the diagnostic criteria of osteoporosis proposed by World Health Organization (WHO). The bone mineral density (BMD) values of lumbar vertebrae and left hips were determined by QDR-2000 dual energy X-ray absorptiometry. Blood samples were tested for plasma biochemical indicators including testosterone, estradiol, calcitonin, osteocalcin, and procollagen type I amino-terminal propeptide by enzyme-linked immunosorbent assay (ELISA), tartrate-resistant acid phosphatase by spectrophotometric method, and the content of nitric oxide by Griess method. Genome DNA was extracted from whole blood, and G894T polymorphism of eNOS gene was analyzed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and 27 bp-VNTR polymorphism of eNOS gene was genotyped by PCR method. Then the relationships between genotypes and biochemical indicators, genotypes and osteoporosis, and haplotypes and osteoporosis were analyzed. Results: The average BMD values of the femoral neck, ward's triangle and lumbar vertebrae 1～4 (L1～L4) in the subjects with T/T genotype in eNOS G894T locus were significantly higher than those in the subjects with G/T and G/G genotypes (P<0.05). The average BMD of the femoral neck in the subjects with a/a genotype of eNOS 27 bp-VNTR locus was evidently higher than that in the subjects with b/b genotype (P<0.05). The plasma testosterone and osteocalcin concentrations in the subjects of eNOS G894T G/T genotype were evidently higher than those in the subjects of other genotypes (P<0.05); the plasma estradiol concentration in the subjects of eNOS 27 bp-VNTR a/a genotype was obviously higher than that in the subjects of b/b genotype (P<0.01). eNOS G/G homozygous frequencies in osteoporosis women, osteopenia women, and normal women were 85.37%, 76.38%, and 83.87%, respectively (P＞0.05). 0% osteoporosis woman, 0.79% osteopenia women, and 3.23% normal women were eNOS a/a homozygous (P<0.05). The frequencies of eNOS 27 bp-VNTR a allele were 5.33% in the osteoporosis group, 10.24% in the osteopenia group, and 16.13% in the normal group (P<0.05, odds ratio (OR)=0.29, 95% confidence interval (CI)=0.11～0.77), suggesting that a/a genotype and a allele might have protective effects on osteoporosis. The haplotype analysis showed that G-b was 87.7% (214/244) in the osteoporosis group (P<0.05, OR=2AS, 95% CI=1.18～5.18). G-a was 5.3% (13/244) in the osteoporosis group (P<0.05, OR=0.29, 95% CI=0.11～0.77). G-b was a risk factor for osteoporosis, and G-a a protective factor. Conclusion: eNOS G894T G/T genotype influenced the plasma testosterone and osteocalcin concentrations, and T/T genotype influenced BMD. eNOS 27 bp-VNTR a/a genotype increased plasma estradiol concentration to have a protective effect on osteoporosis.     

Assignment of <Emphasis Type="Italic">CCR7</Emphasis> gene to chicken chromosome 27 by radiation hybrid panel mapping

The protein encoded by CC chemokine receptor 7 （CCR7） is a member of the G protein-coupled receptor family. This receptor was identified as a gene induced by the Epstein-Barr virus （EBV）, and is thought to be a mediator of EBV effects on B lymphocytes. This receptor is expressed in various lymphoid tissues and activates B and T lymphocytes. It has been shown to control the migration of memory T cells to inflamed tissues, as well as stimulate dendritic cell maturation. To map the CCR7 gene in chicken chromosome, a 6000 rads chicken-hamster radiation hybrid panel （ChickRH6） was used. PCR of samples from ChickRH6 revealed that the location of CCR7 gene is linked to the maker SEQ0347 （6 cR away） with LOD score of 16.6 and that the marker SEQ0347 is located on chromosome 27 at 27 cR of RH （radiation hydrid） map. We compared the corresponding human mRNA sequence with the predicted coding sequence of chicken CCR7 gene, and found that the assembled contig shared a high percentage of similarity with that of the human gene.     

A novel mutation in CD40 ligand gene in a sporadic patient with hyper-IgM syndrome

１ＩｎｔｒｏｄｕｃｔｉｏｎＨｙｐｅｒ－ＩｇＭｓｙｎｄｒｏｍｅ（ＨＩＭ）ｉｓａｒａｒｅｉｍｍｕｎｏｄｅｆｉｃｉｅｎｃｙｃｈａｒａｃｔｅｒｉｚｅｄｂｙａｎｉｎｃｒｅａｓｅｄｓｕｓｃｅｐｔｉｂｉｌｉｔｙｔｏｒｅｃｕｒｒｅｎｔｉｎｆｅｃｔｉｏｎｓａｎｄｍａｒ...     

Anti-angiogenesis effect of generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide on breast cancer in vitro

Objective: To study the effects of the generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide (G4PAMAMNEGFASODN) compound on the expressions of vascular endothelial growth factor (VEGF) and its mRNA of breast cancer cells and on the inhibition of vascular endothelial cells. Methods: We examined the morphology of G4PAMAM/VEGFASODN compound and its pH stability, in vitro transfection efficiency and toxicity, and the expressions of VEGF and its mRNA. Methyl thiazolyl tetrazolium assay was used to detect the inhibitory function of the compound on vascular endothelial cells. Results: The compound was about 10 nm in diameter and was homogeneously netlike. From pH 5 to 10, it showed quite a buffered ability. The 48-h transfection rate in the charge ratio of 1:40 was 98.76%, significantly higher than that of the liposome group (P<0.05). None of the transfection products showed obvious toxicity on the cells. The expressions of both VEGF protein and its mRNA after G4PAMAM/VEGFASODN transfection decreased markedly. Conclusion: With a low toxicity, high safety, and high transfection rate, G4PAMAMNEGFASODN could be a promising gene vector. Specifically, it inhibits VEGF gene expression efficiently, laying a basis for further in vivo animal studies.     

Immunotherapy of intracranial G422 glioblastoma with dendritic cells pulsed with tumor extract or RNA

Objective: To investigate the anti-tumor efficacy of dendritic cell (DC)-based vaccines pulsed with tumor extracts or RNA in a mouse model of intracranial G422 glioblastoma. Methods: Bone marrow-derived DCs were pulsed ex vivo with tumor extracts or RNA. Ninety female mice harboring 4-day-old intracranial G422 glioblastomas and 126 normal mice were treated with three spaced one week apart subcutaneous injections either with PBS, unpulsed DCs, G422 tumor extracts, RNA, DCs pulsed with G422 tumor extracts (DC/extract) or with RNA (DC/RNA). Seven days after the third immunization of normal mice, the spleens of 36 of them were harvested for cytotoxic T lyphocyte (CTL) assays and the others were challenged in the brain with G422 tumor cells. All the treated mice were followed for survival. Some mice brains were removed and examined pathologically when they died. Results: Immunization using DC/extract or DC/RNA significantly induced G422-specific CTL responses compared with control groups (P＜0.01). Vaccination with DC/extract or DC/RNA, either prior to G422 tumor challenge or in tumor-harboring mice, significantly prolonged survival compared with other control groups (P＜0.01). Conclusion: DCs pulsed with tumor extracts or RNA derived from autologous tumors has potential antitumor effects via activation of cell-mediated immunity. Our results suggest a useful therapeutic strategy against gliomas.     

Immunotherapy of intracranial G422 glioblastoma with dendritic cells pulsed with tumor extract or RNA

Objective: To investigate the anti-tumor efficacy of dendritic cell (DC)-based vaccines pulsed with tumor extracts or RNA in a mouse model of intracranial G422 glioblastoma. Methods: Bone marrow-derived DCs were pulsed ex vivo with tumor extracts or RNA. Ninety female mice harboring 4-day-old intracranial G422 glioblastomas and 126 normal mice were treated with three spaced one week apart subcutaneous injections either with PBS, unpulsed DCs, G422 tumor extracts, RNA, DCs pulsed with G422 tumor extracts (DC/extract) or with RNA (DC/RNA). Seven days after the third immunization of normal mice, the spleens of 36 of them were harvested for cytotoxic T lyphocyte (CTL) assays and the others were challenged in the brain with G422 tumor cells. All the treated mice were followed for survival. Some mice brains were removed and examined pathologically when they died. Results: Immunization using DC/extract or DC/RNA significantly induced G422-specific CTL responses compared with control groups (P<0.01). Vaccinatio     

Role of CD97stalk and CD55 as molecular markers for prognosis and therapy of gastric carcinoma patients

Objectives: To explore the mechanism of development and aggressiveness in gastric carcinomas by investigating the expression and role of CD97 and its cellular ligand CD55 in gastric carcinomas. Methods: Tumor and corresponding normal mucosal tissue, collected from 39 gastric carcinoma patients, were examined by immunohistochemistry and RT-PCR for the expression of CD97 and CD55. Results: CD97^stalk was strongly stained on scattered tumor cells or small tumor cell clusters at the invasion front of gastric carcinomas. The expression of CD97^stalk was frequently observed in tumors of stage I and T1 gastric carcinoma patients. The expression of CD97^stalk between Stage I and Stage II, III, IV specimens showed significant difference (P<0.05), between T1 and T2, T3, T4 specimens also showed significant difference (P<0.05). Specimens with tumor invasion depth limited in mucosa of T1 specimens showed higher positive CD55 expression than specimens with the same tumor invasion depth in T2, T3, T4 specimens, the expression of CD55 between T1 and T2 T3, T4 specimens was significantly different (P<0.05). There was strong correlation between the distribution patterns of CD97^stalk and CD55 on tumor tissues (r=0.73,P<0.05). Signet ring cell carcinomas frequently contained strong CD97^stalk and CD55-staining. Conclusions: Our results suggest that CD97^stalk is probably involved in the growth, invasion and aggressiveness of gastric carcinomas by binding its cellular ligand CD55. CD97^stalk and CD55 could be useful as molecular markers for prognosis and therapy of gastric carcinoma patients. Project (No. 2004C34010) supported by the Science and Technology Bureau of Zhejiang Province, China     

Study on interleukin-18 gene transfer into human breast cancer cells to prevent tumorigenicity

INTRODUCTIONInterleukin-18(IL-18),originallydescribedasinterferon(IFN)-g-inducingfactor,isan18.3-kDaproinflammatorycytokineproducedbyactivatedmacrophagesanddendriticcells,andplaysanim-portantroleintheTh1response,primarilybasedonitsabilitytoinduceIFN-gproductioninTcellsandNKcells.IL-18inducesproliferationofactivatedTcells,secretionofseveralcytokines,andpartici-patesinbothinnateandacquiredimmunity.TheroleofIL-18inantitumorimmunitywassuggestedinmanystudies.Genetransferofcytokinesisani…     

Study on interleukin-18 gene transfer into human breast cancer cells to prevent tumorigenicity

To study the effect of interleukin-18 gene transfection on the tumorigenesis of breast cancer cell line Bacp37, human breast cancer cell line Bcap37 were transfected with Lipofectamine and selected by G418. The biological expression of rhIL-18 was tested by RT-PCR and ELISA method; nude mice were injected with Bcap37 cell with or without the hIL-18 gene. The hIL-18 cDNA was successfully integrated into Bcap37 cell; 126.3±4.5 pg hIL-18 secreted by one million transduced cells in 24 hours. Nude mice injected with IL-18 gene engineered Bcap37 cell had no tumor growth. These findings indicated that human breast cancer cells were successfully modified by the gene of IL-18 cytokine; the IL-18 gene engineered Bcap37 cells secreted hIL-18 and lost their tumorigenicity. The Bcap37 cells transduced with IL-18 gene may be used as breast cancer vaccine.     

Effect of perioperative autologous versus allogeneic blood transfusion on the immune system in gastric cancer patients

Background: Allogeneic blood transfusion-induced immunomodulation (TRIM) and its adverse effect on the prognosis of patients treated surgically for cancer remain complex and controversial. However, the potential risk associated with allogeneic blood transfusion has heightened interest in the use of autologous blood transfusion. In the present study, the serum concentrations of neopterin, interferon-gamma (IFN-γ), T lymphocyte subsets (CD3 , CD4 , CD8 , CD4 /CD8 ) and a possible association between these variables were investigated. The purpose was to further evaluate the effect of autologous versus allogeneic blood transfusion on immunological status in patients undergoing surgery for gastric cancer. Methods: Sixty ASA I～II (American Society of Anesthesiologists) patients undergoing elective radical resection for stomach cancer were randomly allocated to receive either allogeneic blood transfusion (n=30) or autologous blood transfusion (n=30). Serum concentrations of the neopterin, IFN-γ and T lymphocyte subsets in the recipients were measured before induction of anesthesia, after operation, and on the 5th postoperative day. Results: Both two groups, serum neopterin, IFN-γ, percentages of T-cell subsets (CD3 , CD4 ), and CD4 /CD8 ratio had significantly decreased after operation, but decreased more significantly in group H (receiving allogeneic blood transfusion) than those in group A (receiving autologous whole blood transfusion) (P<0.05). On the 5th postoperative day, serum neopterin, IFN-γ, CD3 , CD4 T-cells, and CD4 /CD8 ratio returned to the baseline values in group A. In contrast, the above remain decreasing in group H, where there were no significant relations between serum neopterin and IFN-γ. Conclusion: Perioperative surgical trauma and stress have an immunosuppressive impact on gastric cancer patients. Allogeneic blood transfusion exacerbates the impaired immune response. Autologous blood transfusion might be significantly beneficial for immune-compromised patients in the perioperative period, clearly showing its superiority over allogeneic blood transfusion.     

用5＇ LongSAGE标签分析蜜蜂Yps基因转录起始位点的多样性

使用5＇ LongSAGE标签序列确定了一个新的西方蜜蜂Ypsilon Schachtel（Yps）基因的转录起始位点,并进而预测了该基因的启动子序列．5＇ LongSAGE标签的蜜蜂基因组定位结果表明：在成年雄蜂的头部中,蜜蜂Yps基因存在23个转录起始位点,其中优势转录起始位点有4个,其起始频率分别为27．9％、23．1％、13．5％和11．5％．Yps基因TSS的碱基组成分析发现,此23个TSS第一个碱基为A、G、T、C的概率分别为52％、39％、4%、4％．这些结果暗示RNA聚合酶和调控因子在Yps基因启动子的一个较宽范围内的互作控制着不同转录本的起始效率．     

Using chimeric piggyBac transposase to achieve directed interplasmid transposition in silkworm Bombyx mori and fruit fly Drosophila cells

The piggyBac transposon has been long used to integrate foreign DNA into insect genomes. However, undesirable transgene expression can result from random insertions into the genome. In this study, the efficiency of chimeric Gal4-piggyBac transposase in directing integration onto a DNA target plasmid was evaluated in cultured silkworm Bombyx mori Bm-12 and fruit fly Drosophila Schneider 2 (S2) cells. The Gal4-piggyBac transposase has a Gal4 DNA-binding domain (DBD), and the target plasmid has upstream activating sequences (UAS) to which the Gal4 DBD can bind with high affinity. The results indicate that, in the Bm-12 and S2 cells, transpositional activity of Gal4-piggyBac transposase was increased by 4.0 and 7.5 times, respectively, compared to controls, where Gal4-UAS interaction was absent. Moreover, the Gal4-piggyBac transposase had the ability of directing piggyBac element integration to certain sites of the target plasmid, although the target-directing specificity was not as high as expected. The chimeric piggyBac transposase has the potential for use in site-directed transgenesis and gene function research in B. mori.     

Individual Differences in Childhood Behavior Disorders Associated With Epigenetic Modulation of the Cortisol Receptor Gene

Behavioral/emotional difficulties in children are the first sign of mental health problems. These problems are however, heterogeneous. A domain that may identify homogenous subgroups is hypothalamic–pituitary–adrenal function. This study tested whether epigenetic regulation of glucocorticoid receptor gene could explain the co‐occurrence of anxiety problems in children with behavior problems. Four‐ to 16‐year‐old clinically referred children (N = 241) were measured for psychiatric symptoms, methylation of target CpG sites in blood or saliva, and morning cortisol levels in those who gave blood. Increased methylation of promoter 1F CpG sites was associated with higher vulnerability to co‐occurring internalizing symptoms and morning cortisol. The results support increasing optimism that epigenetic regulation of key neuroendocrine systems might help explain hitherto unfathomable individual differences in susceptibility to psychiatric symptom profiles.