人白细胞介素15基因克隆及其在大肠杆菌中的表达

目的 克隆人白细胞介素（IL）15全长cDNA,并在大肠杆菌中表达。方法从人外周血分离的淋巴细胞中提取总RNA,通过RT—PCR扩增出hIL-15全长cDNA。构建到原核表达载体pET28a（＋）中,通过卡那霉素平板筛选及PCR鉴定,挑出阳性克隆进行扩增,DNA测序正确后,转化到大肠杆菌BL21（DE3）中,扩增后IPTG诱导表达。通过SDS-PAEG及Western-blot方法鉴定表达的融合蛋白。结果 成功构建人IL-15表达载体,并表达于大肠杆菌中。SDS—PAEG及Western-blot分析显示表达的融合蛋白分子量为16．1ku,与理论值相符。结论 获得了hIL-15融合蛋白,为hIL-15单克隆抗体的制备及其功能研究奠定了基础。     

Ca15-3在乳腺外Paget病鉴别诊断中的价值

目的：探讨Ｃａ１５３ 在Ｐａｇｅｔ 病鉴别诊断中的价值。方法：对１１ 例阴囊Ｐａｇｅｔ 病回顾性地做Ｃａ１５３、ＥＭＡ、ＣＥＡ和ＨＭＢ４５ 免疫组化染色,并以４ 例Ｂｏｗｅｎ 病和１ 例恶性黑色素瘤做对照。结果：Ｃａ１５３ 、ＥＭＡ 在Ｐａｇｅｔ 病和大、小汗腺及皮脂腺均有较强的表达,４ 例Ｂｏｗｅｎ 病均无Ｃａ１５３ 表达,但其中２ 例有ＥＭＡ 表达。ＣＥＡ 在６ 例（５４．５ ％）Ｐａｇｅｔ 病中有表达。ＨＭＢ４５ 只在恶性黑色素瘤中表达。结论：Ｃａ１５３ 在Ｐａｇｅｔ 细胞的相对特异性表达提示它可作为Ｐａｇｅｔ 病与Ｂｏｗｅｎ 病鉴别诊断的标志物,其对诊断的敏感性与ＥＭＡ 相同,但特异性高于ＥＭＡ。同时还说明Ｐａｇｅｔ 病的发生可能与汗腺有关。     

原发胃癌中p15、p16基因蛋白的表达及临床意义

目的检测原发胃癌原发灶及癌旁正常组织中p15、p16基因蛋白表达情况,探讨p15、p16基因失活与胃癌发生发展及临床病理特征的关系。方法应用免疫组织化学方法检测32例进展期胃癌石蜡标本中p15、p16的表达,并探讨其表达与临床病理特征之间的关系。结果p16、p15在原发灶与癌旁正常组织中阳性表达率差异有统计学意义（P〈0．05）,p15、p16基因表达在有无淋巴结转移和,下同TNM分期的病例之间差异均有统计学意义（P〈0．05）。结论p15、p16蛋白袁达可能与胃癌癌变过程相关．并可以作为临床治疗提供理论依据。     

Hirschsprung-associated enterocolitis develops independently of NOD2 variants

Backgroud/Purpose

Hirschsprung-associated enterocolitis (HAEC) represents a cause for significant pre- and postoperative morbidity and mortality in Hirschsprung disease (HD). Although multiple studies on HAEC have been performed and several mechanisms have been presumed, the pathogenesis of this condition remains unclear. As changes in colonic mucosal defense are key factors suggested in both Crohn's disease (CD) and HAEC pathogenesis, the aim of the current study was to investigate genetic alterations in the most important susceptibility gene for Crohn's enterocolitis (NOD2) to see whether carriers of polymorphisms within the NOD2 gene are predisposed to the development of HAEC.

Methods

Genotyping for the NOD2 variants in exon 4 (p.Arg702Trp [rs2066844]), exon 8 (p.Gly908Arg [rs2066845]), and exon 11 (p.1007fs [rs2066847]) was performed in 52 white children with HD (41 boys, 11 girls), 152 healthy controls, and 152 children with CD (onset of disease <17 years; mean, 11.8 years). Seventeen patients with HD (32.7%) were carriers of a RET germline mutation, 35 children (67.3%) had short segment disease, and 17 (32.7%) had long segment disease.

Results

Ten children (19.2%) with HD were heterozygous carriers of at least one NOD2 variant vs 17 (11.2%) in the healthy control group and 69 (45.4%) in the CD cohort. Hirschsprung-associated enterocolitis was observed in 7 children (13.5%), with 4 having short segment HD and 3 with long segment HD; but none of them were carriers of NOD2 variants.

Conclusion

Our study shows that NOD2 variants described to be causatively associated with CD do not predispose to the development of HAEC. As data on the molecular basis of HAEC are limited, the distinct mechanisms involved in the pathogenesis of this complication remain unclear.     

TLR7激动剂Gardiquimod上调BxPC-3细胞中IL-15 mRNA表达

目的：研究TLR7在胰腺癌BxPC-3细胞中表达,并探讨TLR7激动剂激活TLR7后细胞因子白介素15( IL-15)的表达。方法通过 Western blot 和 Real-time PCR 分析TLR7在细胞内的表达水平;BxPC-3细胞经过不同时间点Gardiquimod(3μg/ml)的处理后,Real-time PCR 分析 TLR7激活后 IL-15 mRNA 水平表达变化;Western blot 分析Gardiquimod刺激细胞后,磷脂酰肌醇-3-激酶-蛋白质丝氨酸苏氨酸激酶( PI3K-AKT)信号通路的变化。结果 Western blot和 Real-time PCR结果显示：与外周血单核细胞( PBMC)相比, TLR7在 BxPC-3中弱表达;Real-time PCR分析显示：Gardiquimod处理细胞后能刺激细胞中IL-15的表达;TLR7激动剂能够激活 PI3K-AKT信号通路。结论 Gardiquimod激活TLR7后能够上调 IL-15的表达,并且其激活与 PI3K-AKT信号途径相关。     

乳腺肿块患者血清CA15-3及CEA检测的临床意义

目的 探讨电化学发光法检测血清糖类抗原15-3(CA15-3)及癌胚抗原(CEA)在乳腺疾病诊断以及术后监测中的临床应用价值.方法 分析2005年5月至2006年12月间住院患者血清CA15-3及CEA检测结果,结合临床病历资料进行统计学分析.结果 乳腺良性肿块组与乳腺癌组CA15-3及CEA检测结果差异显著,血清CA15-3及CEA用于诊断乳腺癌的特异性较高(97.6%及96%),但敏感性较低(30.5%及13.9%);血清CA15-3异常的乳腺癌患者,术后CA15-3明显下降;CA15-3与CEA之间存在一定的相关性,但相关系数不高.结论 CA15-3及CEA在诊断早期乳腺癌时虽然特异性很高,但由于敏感性太低导致其临床应用价值有限,二者联合应用对提高诊断效率意义不大,当乳腺癌患者血清CA15-3较高时,可以用于评价治疗效果.对于乳腺癌的早期诊断,尚需寻找更敏感的标志物.     

A novel recessive 15-hydroxyprostaglandin dehydrogenase mutation in a family with primary hypertrophic osteoarthropathy

Abstract

We present two PHO siblings having a novel homozygous truncating mutation in HPGD. The purpose of the study was to attempt medical treatment, and to find the HPGD mutation causing the disease, in a 22-year old Turkish male and his 23-year old sister afflicted with primary hypertrophic osteoarthropathy (PHO). In combination with NSAIDs and colchicine, treatment with sulfasalazine was started in both cases, and methotrexate was added to the treatment regimen of the female patient at the end of the first year. The patients were found to be typical PHO. Ultrasonographic examination of the joints revealed synovitis and inflammation by B mode and power Doppler ultrasonography. Joint symptoms responded to sulfasalazine treatment in both patients. However, after the addition of methotrexate, the female patient had better remission. All exons of HPGD, the known disease gene, were analyzed by Sanger sequencing. A homozygous 2-bp deletion (c.310_311delCT or p.L104AfsX3) was identified. Seven relatives carrying the mutation in the heterozygous state were examined and none was found affected. Although not specific for this disease, skin, soft tissue and joint ultrasonography can be helpful for evaluation of the musculoskeletal findings in the patients.     

Ordered mesoporous silica modified with lanthanum for ibuprofen loading and release behaviour

The ordered mesoporous silicas SBA-15 and KIT-6, modified with lanthanum, have been for the first time applied in investigation of ibuprofen adsorption and release. The materials of hexagonal and regular structure were obtained by the hydrothermal method using a triblock copolymer Pluronic P123 as a template. The mesoporous silicas were impregnated with an aqueous solution of lanthanum(III) chloride in the amount necessary to obtain 1, 3 and 5 wt.% La loading. The physicochemical properties of the modified silicas were characterised by X-ray diffraction, transmission electron microscopy, UV–Vis spectrophotometry and low-temperature nitrogen sorption. The results showed that lanthanum strongly determined structural as well as textural properties of the silicas. The samples of modified silica were checked for the ability to adsorb and release of ibuprofen. The storage capacity of the modified silicas obtained increased with increasing their average pore diameter and percentage content of lanthanum. The amount of ibuprofen adsorbed onto KIT-6 silica modified with La was higher than that adsorbed onto SBA-15 materials. The high coverage of lanthanum on the surface of KIT-6 and SBA-15 solids was found to increase the amount of ibuprofen and the rate of its release.