凋亡相关基因 bcl-2、bax、bad与乳腺癌

目的探讨乳腺癌发生、发展过程中bcl-2、bax及bad基因表达水平变化及与乳腺癌其他生物因素的相关性。方法复习国内、外相关文献并进行综述。结果在从正常乳腺组织到乳腺癌逐渐演化的过程中凋亡抑制基因bcl-2的表达水平变化尚有待进一步研究,而凋亡促进基因bax、bad的表达水平呈递减趋势。bcl-2基因表达水平与乳腺癌中一些公认的正性因子如雌激素受体(ER)、孕激素受体(PR)呈正相关,与其他一些负性因子如p53、表皮生长因子受体、c-erbB-2、腋窝淋巴结转移情况等呈负相关。bax基因的表达水平与乳腺癌ER、PR水平以及p53表达水平等无关。对于bad基因与乳腺癌其他生物因素的相关性尚未见报道。结论乳腺癌中bcl-2基因表达水平的变化对乳腺癌预后以及治疗的作用尚存在争议,bad基因与乳腺癌预后的关系亦不清楚,而bax基因表达水平与乳腺癌的预后呈正相关。对于它们的研究将为我们评价乳腺癌的预后提供新的指标,为乳腺癌的治疗开辟新的思路。     

睾丸特异性基因TSF22在小鼠中的表达研究

目的 筛选与精子发生相关的基因。方法 将4、9、18、35、54日龄和6月龄小鼠睾丸组织cDNA探针与Affymetrix全基因组芯片进行杂交，筛选出差异表达的基因。利用网络信息资源对该基因进行生物学信息分析。RT-PCR分析该基因在小鼠睾丸不同发育阶段及小鼠不同组织中的表达。结果 芯片结果分析筛选出一个差异表达杂交点（GenBank登录号：NM_199034），生物信息学分析发现该基因全长1597bp，含有570bp的完整ORF，编码190个氨基酸、分子量为22．106kDa的蛋白质，我们将其命名为TSF22。RT-PCR分析表明TSF22基因特异性表达于睾丸组织及18日龄小鼠睾丸中。结论 TSF22基因的表达与小鼠精子发生的过程一致，可能在精子发生中起重要作用。     

骨肉瘤组织中p16基因的缺失突变及CDK4基因的扩增

目的 确定在原发性骨肉瘤中是否有Ｐ１６基因的异常及Ｐ１６基因异常与ＣＤＫ基因扩增之间的关系。方法 应用定量Ｓｏｕｔｈｅｒｎ ｂｌｏｔ方法分析确定Ｐ１６基因的缺失及ＣＤＫ基因的扩增。用ＰＣＲ＝ＳＳＣＰ方法检测Ｐ１６基因突变,结果：在６０例分析Ｐ１６基因的标本中有５例（９％）存在Ｐ１６基因的重排或缺失。均发生于原发肿瘤。在６７例分析ＣＤＫ４基因的标本中,有６例（９％）存在ＣＤＫ４基因的扩是有包括３例,     

β-连环蛋白及APC基因与肿瘤

目的：探讨β－连环蛋白和结肠腺瘤性息肉病（APC）基因的分子结构和功能特征以及其在肿瘤发生、发展的作用。方法：综述近年来有关β－连环蛋白和APC基因分子生物学以及与肿瘤学方面的研究。结果：β－连环蛋白和APC基因由于自身调节和相互调节异常，在肿瘤发生的早期表达异常增高，但与肿瘤的进展、转移关系似乎正相反。此外，β－连环蛋白和APC基因还可以调节p53、c-myc基因以及细胞周期蛋白D1等因子表达。结论：β－连环蛋白和APC基因在多种肿瘤的发生、发展中可能起着中心作用，并与其它癌／抑癌基因和因子相互产生调节作用。     

金黄色葡萄球菌耐药性及mecA、qacA／B和pvl基因检测

金黄色葡萄球菌是临床上最常见的致病菌之一,可产生多种毒素和酶,包括凝固酶、葡萄球菌溶血素、杀白细胞素、肠毒素和表皮溶解毒素等。耐甲氧西林金黄色葡萄球菌（MRSA）具有多药耐药特征,是导致医院感染的主要革兰阳性菌,已成为临床抗感染治疗的难题。监测金黄色葡萄球菌的耐药现状,了解金黄色葡萄球菌的耐药和致病机制,以及分析感染的危险因素与治疗对策具有重要的临床意义。     

IMMUNOTHERAPY,INCLUDING GENE THERAPY,FOR METASTATIC MELANOMA

Current standard therapy for distant metastatic melanoma is ineffective and often compromises the quality of a patient's life. Immunotherapy is briefly reviewed in relation to its many forms: from local non-specific to the more recent specific vaccines, including those using specific melanoma peptides (e.g. from the proteins encoded by melanoma-associated gene (MAGE)) and those involving genetically transduced autologous melanoma cells using retroviral vectors in vitro. The mode of action of genetically transduced melanoma cells incorporating the granulocyte macrophage colony stimulating factor (GM-CSF) gene (GVAX) is presented as a paradigm for cytokine-mediated strategies. Trials of GVAX and other cytokine gene strategies are under way in Brisbane, Boston and Amsterdam, and some interim perspectives on the clinical outcomes and immunological mechanisms involved are sketched. Some of the compounding problems in immunotherapeutic strategies for cancer are identified, and possible adjunct manoeuvres for overcoming them are discussed.     

Localization of APOL1 Protein and mRNA in the Human Kidney: Nondiseased Tissue,Primary Cells,and Immortalized Cell Lines

Although APOL1 gene variants are associated with nephropathy in African Americans, little is known about APOL1 protein synthesis, uptake, and localization in kidney cells. To address these questions, we examined APOL1 protein and mRNA localization in human kidney and human kidney-derived cell lines. Indirect immunofluorescence microscopy performed on nondiseased nephrectomy cryosections from persons with normal kidney function revealed that APOL1 protein was markedly enriched in podocytes (colocalized with synaptopodin and Wilms’ tumor suppressor) and present in lower abundance in renal tubule cells. Fluorescence in situ hybridization detected APOL1 mRNA in glomeruli (podocytes and endothelial cells) and tubules, consistent with endogenous synthesis in these cell types. When these analyses were extended to renal-derived cell lines, quantitative RT-PCR did not detect APOL1 mRNA in human mesangial cells; however, abundant levels of APOL1 mRNA were observed in proximal tubule cells and glomerular endothelial cells, with lower expression in podocytes. Western blot analysis revealed corresponding levels of APOL1 protein in these cell lines. To explain the apparent discrepancy between the marked abundance of APOL1 protein in kidney podocytes observed in cryosections versus the lesser abundance in podocyte cell lines, we explored APOL1 cellular uptake. APOL1 protein was taken up readily by human podocytes in vitro but was not taken up efficiently by mesangial cells, glomerular endothelial cells, or proximal tubule cells. We hypothesize that the higher levels of APOL1 protein in human cryosectioned podocytes may reflect both endogenous protein synthesis and APOL1 uptake from the circulation or glomerular filtrate.     

整合素α6、β-微精浆蛋白基因和染色体8q24区与前列腺癌的关联研究

目的 探讨整合素α6(ITGA6)基因(rs12621278,G)、染色体8q24区(rs10086908,T)和β-微精浆蛋白(MSMB)基因(rs10993994,T)与北京市居民中前列腺癌(PCa)的关联,了解PCa患者基因型和表型的关系.方法 收集112例PCa患者临床、遗传、膳食习惯、嗜好等表型,对PCa患者和91名正常对照者的ITGA6基因(rs12621278,G)、染色体8q24区(rs10086908,T)和MSMB基因(rs10993994,T)进行比较,并进行病例组的基因型-表型分析.结果 2组间相比,MSMB基因rs10993994,T风险等位基因频率差异有统计学意义(病例组56.4%,对照组46.2%;P=0.001,OR=1.97,95%CI为1.28～3.04).8q24区的rs10086908,T(病例组83.5%,对照组79.2%)和ITGA6基因的rs12621278,G(病例组27.2%,对照组27.0%)组间差异无统计学意义(P＞0.05).数量性状分析发现ITGA6风险基因型(G/G)的患者病程为(1.40±0.55)年,显著短于A/G型的(4.38±3.10)年和A/A型的(2.37±1.84)年(P=0.003).结论 MSMB基因变异和PCa易感性之间存在相关性,提示MSMB基因可能与PCa有关联.

Abstract：

Objective To explore the correlation between ITGA6 gene (rs12621278, G), MSMB gene (rs10993994, T), chromosome 8q24 (rs10086908, T) and prostate cancer (PCa) in Beijing residents, and to explore the correlation between genotype and phenotype in PCa patients. Methods PCa patient phenotypes were collected including clinical, genetic, dietary habits, hobbies and blood samples. ITGA6 gene (rs12621278, G), chromosome 8q24 (rs10086908, T) and MSMB gene (rs10993994, T) compared the allele distribution between 112 PCa and 91 healthy control age matched patients. The genotype and phenotype analysis was conducted in the 2 groups. Results Between the case and control groups, only rs10993994, T of MSMB gene (case 56.4%,control 46.2%) was significantly different (P=0.001; OR=1.97, 95%CI:1.28-3.04). The rs10086908, T of 8q24 (case 83.5%, control 79.2%) and rs12621278, G of ITGA6 gene (case 27.2%, control 27.0%) were not significantly associated with PCa in the study sample (P>0.05). Quantitative trait analysis showed that the disease duration of ITGA6 risk genotypes (G/G,1.40±0.55 years) in PCa patients was significantly shorter (P=0.003) than the other genotype carriers (A/G, 4.38±3.10 years; A/A, 2.37±1.84 years). Conclusion The genetic variation in MSMB is possibly associated with PCa susceptibility, suggesting that MSMB genes might be associated with PCa in a Chinese population.     

P-gp、nm23和p53联合检测判断乳腺癌预后的价值

目的　研究乳腺癌组织中P gp、nm23和p53的变化规律及与乳腺癌复发的关系。方法　应用免疫组化方法检测10例乳腺纤维腺瘤(纤维腺瘤组)和47例乳腺癌病变组织中P gp、nm23和p53 的表达,47例乳腺癌中18例随访3~5年无复发(无复发组),29例3年内复发(复发组),并比较其表达阳性率和表达强度的差异。结果　乳腺癌复发组nm23表达阳性率较纤维腺瘤组低(P<0.05),其阳性表达强度较无复发组低。复发组和无复发组p53表达阳性率均较纤维腺瘤组高(P<0.05),复发组p53阳性表达强度高于无复发组(P<0.05)。复发组和无复发组P gp表达阳性率与纤维腺瘤组比较差异无显著性意义,但其阳性表达强度较纤维腺瘤组低(P<0.01,P<0.025)。结论　p53过表达和nm23低表达对预测乳腺癌复发具有一定价值。     

5例无精症病人染色体核型及YRRM1、DYS240基因检测

目的：探讨无精症病人的发病原因。方法：对4例Y染色体有结构畸变、1例46，XX男性无精症病人及10例正常有生育力的男性用PCR方法进行YRRM1、DYS240基囚的检测。结果：10例有生育力核型正常的男性及2例inv（Y）病人均检出YRRM1和DYS240基因片段，而2例del（Y）病人未能检出YRRM1及DYS240基因片段，但他们的父亲均有正常的染色体及YRRM1和DYS基因片段，46，XX男性经检测证实有SRY基囚的存在而无YRRM1和DYS240基因。结论：以上结果表明，发生在Y染色体长臂异染色质区及短臂末端的倒位不会引起YRRM1、DYS240基因的缺失，而Y染色体长臂q11以远部位的缺失则都有YRRM1和DYS240基因的缺失，是造成无精症的原因。     

Gene analysis，treatment,and follow-up of sixteen Chinese patients with Bartter syndrome

Objective To analyze the mutations of causal genes in sixteen Chinese patients with suspicious Bartter syndrome, and follow up their treatment results. Methods Mutations were identified by the next generation sequencing and the multiplex ligation-dependent probe amplification (MLPA). Clinical and biochemical features at the first presentation as well as follow-up results were reviewed. Results 15 different CLCNKB gene mutations were identified in sixteen patients with BS, including 11 novel ones. A novel missense mutation and a novel small deletion were found from SLC12A1 gene. A novel gross deletion was found in CLCNKA gene. A recurrent missense mutation was identified from BSND gene. The whole gene deletion mutation of CLCNKB gene was the most frequent mutation (32%), and the rate of gross deletion was up to 50 percent in this group of Chinese patients. The most common clinical manifestations were development retardation (15/16), polydipsia and polyuria (15/16). All of the patients were detected with hypokalemia, hypochloremia and metabolic alkalosis. Indomethacin treatment had significant improvement to the stature and weight restoration. Conclusion The present study has found 19 mutations, including 14 novel ones, which enriches the human gene mutation database (HGMD) and provides valuable references to the genetic counseling and diagnosis of Chinese population.     

抑癌基因P16与癌基因P21在膀胱肿瘤中的表达及意义

目的：探讨癌基因Ｐ２１和抑癌基因Ｐ１６的表达与膀胱癌生物学行为之间的关系。方法：采用免疫组织化学Ｓ－Ｐ法对４０例膀胱移行细胞癌癌基因Ｐ２１和抑癌基因Ｐ１６进行检测。结果：Ｐ２１和Ｐ１６阳性表达率为６７．５％和５２．５％;Ｐ２１阳性表达率与膀胱移行细胞癌的病理分级和临床分期呈正相关,而Ｐ１６阳性表达率与膀胱移行细胞癌的病理分级和临床分期呈负相关。结论：Ｐ２１、Ｐ１６基因蛋白的表达与膀胱肿瘤组织病理分     

Single‐nucleotide polymorphism in Turkish patients with adolescent idiopathic scoliosis: Curve progression is not related with MATN‐1, LCT C/T‐13910, and VDR BsmI

The role of genetics in the etiopathogenesis of adolescent idiopathic scoliosis (AIS) is unclear. In this study, we investigated the relationship between AIS and polymorphisms in MATN‐1, LCT C/T‐13910, and VDR BsmI genes. 53 Turkish adolescents with diagnosed AIS and 54 healthy adult individuals were included in the study. MATN‐1, LCT C/T‐13910, and VDR BsmI gene mutations were analyzed with real‐time PCR. We did not detect a statistically significant difference between AIS and control groups in respect to those three different gene polymorphisms (p < 0.05). We next evaluated the associations of all three SNPs with scoliosis curve severity. There was no significant difference between curve severity and gene polymorphisms (p < 0.05). In terms of gene polymorphisms, AIS patients with a family history of AIS did not significantly differ from AIS patients who did not have history (p < 0.05). AIS might be caused by many different gene mutations, biomechanical mechanisms that have been modified by environmental factors, different biological interactions, modulation of growth, or a synergy of different factors causing abnormal control of growth. However, the existing knowledge is still not enough to explain the etiopathogenesis of AIS. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1459–1463, 2012     

Polymorphisms in the genes encoding the 4 RET ligands, GDNF, NTN, ARTN, PSPN, and susceptibility to Hirschsprung disease

Purpose

Hirschsprung disease (HSCR) is a developmental disorder caused by a failure of neural crest cells to migrate, proliferate, and/or differentiate during the enteric nervous system development. It presents a multifactorial, nonmendelian pattern of inheritance, with several genes playing some role in its pathogenesis. Its major susceptibility gene is the RET protooncogene, which encodes a receptor tyrosine kinase activating several key signaling pathways in the enteric nervous system development. Given the pivotal role of RET in HSCR, the genes encoding their ligands (GDNF, NRTN, ARTN, and PSPN) are also good candidates for the disease.

Methods

We have performed a case-control study using Taqman technology to evaluate 10 polymorphisms within these genes, as well as haplotypes comprising them, as susceptibility factors for HSCR.

Results

No differences were found in the allelic frequencies of the variants or in the haplotype distribution between patients and controls. In addition, no particular association was detected of the variants/haplotypes to any demographic/clinical parameters within the group of patients.

Conclusion

These data would be consistent with the lack of association between these polymorphisms and HSCR, although they do not permit to completely discard a possible role of other variants within these genes in the disease. Moreover, because the gene-by-gene approach does not take into account the polygenic nature of HSCR disease, it would be interesting to investigate sets of variants in many other different susceptibility loci described for HSCR, which may permit to consider possible interactions among susceptibility genes     

瘢痕疙瘩Fas基因外显子6,8,9的突变分析

目的：研究瘢痕疙瘩Fas基因中外显子6，8，9的突变情况，了解该基因突变与瘢痕疙瘩形成之间的关系。方法：实验分正常皮肤、增生性瘢痕和瘢痕疙瘩组织三组，采用PCR-SSCP(聚合酶链反应-单链构象多态性)方法筛选突变，然后进行序列分析以确定突变。结果：经银染SSCP分析，在23例瘢痕疙瘩标本中，出现异常电泳带的有18例，经测序，外显子6，8，9突变分别有8例、12例和7例，6和8同时存在突变的有2例，6和9同时存在突变的有3例，8和9同时存在突变的有2例，三个外显子同时有突变的有l例，而且发现了新的突变，在正常皮肤组织和增生性瘢痕组织中均未检出突变。结论：瘢痕疙瘩组织中Fas基因的突变可能与该病的发生有密切关系；PCR-SSCP是检测基因点突变的一种快速、敏感、有效的方法。     

胆囊癌组织细菌L型的检测与p21及p53基因突变的关系

目的 探讨胆囊癌的发生与细菌L型感染和p21及p53基因突变的关系。方法 采用免疫组化SP法和革兰氏染色技术，对40例胆囊癌及40例慢性胆囊炎组织中的细菌L型和突变型p53蛋白及p21蛋白进行检测，并对胆囊癌组织中的细菌L型阳性伴p21及p53蛋白阳性表达结果和细菌L型阴性伴p21及p53蛋白阳性表达结果进行对比分析。结果 胆囊癌组织中革兰氏染色细菌L型检出率为77．5％(31／40)，明显高于胆囊炎的57．5％(23／40)，P＜0．05，且与免疫组化细菌L型抗原表达阳性率[80．0％(32／40)]具有一致性；胆囊癌组织中的p21及p53蛋白表达阳性率分别为62．5％(25／40)和65．0％(26／40)，明显高于胆囊炎组织中的p21及p53蛋白表达阳性率[2．5％(1／40)和5.0％(2／40)]，P＜0．05；胆囊癌组织中的细菌L型阳性者其p21及p53蛋白表达阳性率分别为75．0％(24／32)和78．1％(25／32)，明显高于细菌L型阴性者的p21及p53蛋白的表达阳性串率[12.5％(1／8)，12．5％(1／8)]，P＜0。05。结论 细菌L型参与了p21及p53基因的突变，并在胆囊癌的发生中可能起协同作用。     

Functional genomics in osteoarthritis: Past,present, and future

Osteoarthritis (OA) is a common complex disease of high public health burden. OA is characterized by the degeneration of affected joints leading to pain and reduced mobility. Over the last few years, several studies have focused on the genomic changes underpinning OA. Here, we provide a comprehensive overview of genome‐wide, non‐hypothesis‐driven functional genomics (methylation, gene, and protein expression) studies of knee and hip OA in humans. Individual studies have generally been limited in sample size and hence power, and have differed in their approaches; nonetheless, some common themes have started to emerge, notably the role played by biological processes related to the extracellular matrix, immune response, the WNT pathway, angiogenesis, and skeletal development. Larger‐scale studies and streamlined, robust methodologies will be needed to further elucidate the biological etiology of OA going forward. © 2016 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 34:1105–1110, 2016.     

Determination of annulus fibrosus cell response to tensile strain as a function of duration,magnitude, and frequency

As clinical evidence suggests that mechanical forces can have both reparative and traumatic effects on the spine, we investigated responses to different magnitudes, frequencies, and durations of applied tensile strain in an in vitro system. We examined the interactions of inflammatory and mechanical stimuli on cells isolated from the annulus fibrosus. Rabbit annulus fibrosus fibrochondrocytes were cultured in the presence or absence of an inflammatory stimulus. Cells were exposed to various magnitudes and frequencies of tensile strain for 4 or 24 h, and mRNA expression of catabolic mediators of inflammation and matrix degradation was measured by quantitative real time PCR and compared to control cells. Conditioned media were analyzed for matrix metalloprotease activity and production of prostaglandin E₂. Application of low magnitudes and frequencies of tensile strain resulted in down‐regulation of catabolic mediators, particularly under inflammatory stress. However, loss of this protective effect was observed at higher frequency and magnitude, and after prolonged duration. These in vitro data confirm the existence of magnitude, frequency, and duration based effects, which determine biochemical response of disc tissue resulting in either anti‐ or pro‐catabolic outcomes. This may help to explain the beneficial effects of motion‐based therapies as well as the destructive effect of traumatic levels of applied strain. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29: 1275–1283, 2011