Proteases and their inhibitors as prognostic factors for high-grade serous ovarian cancer

Ovarian carcinoma is one of the most lethal malignancies, but only very few prognostic biomarkers are known. The degradome, comprising proteases, protease non-proteolytic homologues and inhibitors, have been involved in the prognosis of many cancer types, including ovarian carcinoma. The prognostic significance of the whole degradome family has not been specifically studied in high-grade serous ovarian cancer. A targeted DNA microarray known as the CLIP-CHIP microarray was used to identify potential prognostic factors in ten high-grade serous ovarian cancer women who had early recurrence (<1.6 years) or late/no recurrence after first line surgery and chemotherapy. In women with early recurrence, we identified seven upregulated genes (TMPRSS4, MASP1/3, SPC18, PSMB1, IGFBP2, CFI – encoding Complement Factor I – and MMP9) and one down-regulated gene (ADAM-10). Using immunohistochemistry, we evaluated the prognostic effect of these 8 candidate genes in an independent cohort of 112 high-grade serous ovarian cancer women. Outcomes were progression, defined according to CA-125 criteria, and death. Multivariate Cox proportional hazard regression models were done to estimate the associations between each protein and each outcome. High ADAM-10 expression (intensity of 2–3) was associated with a lower risk of progression (adjusted hazard ratio (HR): 0.51; 95% confidence interval (CI): 0.29-0.87). High complement factor I expression (intensity 2–3) was associated with a higher risk of progression (adjusted HR: 2.30, 95% CI: 1.17–4.53) and death (adjusted HR: 3.42; 95% CI: 1.72–6.79). Overall, we identified the prognostic value of two proteases, ADAM-10 and complement factor I, for high-grade serous ovarian cancer which could have clinical significance.     

外周血液中miRNA表达与老年性黄斑变性相关性研究

目的　检测微小核糖核酸（microribonucleicacids，miRNA）在老年性黄斑变性（age-related macular degeneration，AMD）患者中的表达，并探讨miRNA的表达量与AMD病程之间的关系。方法　选取2014年1月至2016年11月于同济大学附属第十人民医院眼科门诊就诊的AMD患者6例为试验组，并选取同期6名正常人为对照组，通过基因芯片技术检测两组血液中miRNA的表达量。扩大样本的病例对照研究中共纳入126例AMD患者和140名正常人，检测其血液样本中miRNA的表达，比较两组人群间miRNA的表达量差异。结果　通过基因芯片技术，在试验组与对照组间共检测出216个miRNA存在表达差异（均为P<0.05），与对照组相比，试验组中111个miRNA表达量上升，105个miRNA表达量下降，差异均有统计学意义（均为P<0.05）。扩大样本的病例对照研究结果表明，在AMD患者中，miR-27a-3p、miR-29b-3p、miR-195-5p的表达量显著上升，同时，湿性AMD患者血液中miR-27a-3p的表达量高于干性AMD患者，差异均有统计学意义（均为P<0.05）。结论　AMD患者外周血中miRNA表达量水平有明显变化，miR-27a-3p、miR-29b-3p、miR-195-5p可能成为AMD血清学诊断和预后的标志物。     

A novel lncRNA-LINC01116 regulates tumorigenesis of glioma by targeting VEGFA

Brain glioma is the most common malignant tumor of the central nervous system, and one of the leading causes of death in patients with intracranial tumors. The clinical outcome of glioma is usually poor due to abundant vascularity, fast growth and susceptibility of invasion to normal brain tissues. Our microarray study showed that lncRNA-LINC01116 was significantly upregulated in glioma tissues and played an important role in cell proliferation, cycle, migration, invasion and angiogenesis. In addition, vascular endothelial growth factor (VEGFA) may be the major target genes in the downstream of lncRNA-LINC01116. Dual luciferase assay showed that LINC01116 and VEGFA both contained a miR-31-5p binding site, and LINC01116 could regulate the expression of VEGFA through competitive absorption of miR-31-5p. RNA immunoprecipitation indicated that LINC01116 and VEGFA were present in the miR-31-5p-RISC complex, and biotinylated miR-31-5p pull-down assay suggested that there was a competitive relationship between LINC01116 and VEGFA to bind with miR-31-5p. Collectively, our study has identified a novel lncRNA-LINC01116 and clarified the role and mechanism of LINC01116 in the tumorigenesis of glioma. LINC01116 may prove to be a potential target for the clinical diagnosis and treatment of glioma.     

一例外周血染色体核型为单纯型18三体但智力正常女性的遗传学研究

目的分析1例外周血染色体核型为单纯型18三体但智力正常女性的遗传学机制。方法用G显带染色体核型分析、荧光原位杂交(fluorescence in situ hybridization,FISH)和单核苷酸多态性微阵列芯片(single nucleotide polymorphism microarray,SNP-array)技术对患者的外周血和颊粘膜细胞进行检测。结果患者的外周血染色体核型、SNP-array以及FISH检测结果均提示为47,XX,+18;颊粘膜间期细胞FISH检测结果提示为45,X合并低比例的18三体和18单体。结论胚层染色体嵌合的个体临床表现复杂,遗传学异常所造成的影响取决于相关胚层分化所形成的器官及功能。     

糖尿病肛瘘lncRNA芯片分析及CNC图搭建＊

目的 研究糖尿病肛瘘创面特异表达LncRNA与mRNA基因功能之间调控网络。方法 用基因芯片技术对糖尿病肛瘘创面和普通肛瘘创面组织中差异性表达的LncRNA及mRNA进行基因表达谱检测，筛选的标准为2倍差异及P < 0.05,再进行差异表达分析，然后对差异表达的mRNAs进行KEGG通路分析及Pathway Map展示，挑选出显著mRNA指标，将这些显著mRNA指标进行q-PCR验证，得到有意义的阳性指标，再将阳性指标与差异LncRNA交集得出LncRNA-mRNA共表达网络，并基于LncRNA-mRNA共表达网络挑选出不同LncRNA进行功能验证。结果 将芯片标准化后分析差异表达的长链非编码RNA和mRNA，发现上调差异有502个，下调差异有1204个；mRNA分析发现上调差异621个，下调差异505个，KEGG通路分析发现上调和下调的通路均有10条；通过分别对上调、下调最明显的通路进行Pathway Map展示，挑选出显著mRNA指标8个：BMP2、IFNB1、IL6、IL18、PIK3CB、SMAD7、SMAD9、β-actin，分别将其进行q-PCR验证，得到有意义的阳性指标个5个：BMP2、IL6、IL18、PIK3CB、SMAD7，将5个阳性指标和差异LncRNA交集得出LncRNA-mRNA共表达网络，并基于LncRNA-mRNA共表达网络挑选出不同LncRNA进行功能验证。结论 挑选出的20个LncRNA（NR_125383、T323486、ENST00000582334、TCONS_00018312、ENST00000418393、TCONS_00017190、TCONS_00019532、ENST00000601559、ENST00000566575、NR_109882、NR_026913、T301537、NR_109774、ENST00000415536、ENST00000610000、ENST00000412485、ENST00000573220、T175957、ENST00000580756、TCONS_00014747）所调控的mRNA居于LncRNA-mRNA共表达网络，其在各个领域当中均有不同程度的报道，为后续的功能和机制研究提供了方向和重点，为加速慢性难愈合和创面愈合的研究提供了新的思路，为开发促愈药物提供基础研究借鉴。     

Improved Prognostication for the Updated AJCC Breast Cancer Pathological Prognostic Staging Varied in Higher-Stage Groups

BackgroundIn addition to TNM-based anatomical staging (AS), a novel pathological prognostic staging (PPS) has been proposed by the American Joint Committee on Cancer (AJCC). PPS demonstrated better prognostication, but its superiority in breast cancer subtypes and related to staging discrepancies between AS and PPS are not clear.MethodsA cohort of 1729 patients with breast cancer was staged into AS and PPS according to the latest AJCC staging. Patient characteristic and restaging outcomes were compared.ResultsCompared with AS, 799 and 135 cases were upstaged and downstaged respectively in PPS, mostly involved stage I cases. For the overall cohort, PPS demonstrated superior prognostic power over AS in both disease-free survival (DFS) and breast cancer–specific survival. However, such superiority was found mainly in estrogen receptor (ER)/progesterone receptor (PR)+ but not ER−PR− cancers. Comparing the restaged cases within the same PPS, PPS 1A cases showed similar survival irrespective of the original AS. Interestingly, in other PPS groups (PPS 1B and higher), there was a difference in outcome among patients with same PPS but different AS. Within PPS 1B patients, downstaged cases from higher AS showed worse DFS (3A>1B vs. 2A>1B: χ² = 4.732, P = .030).ConclusionsPPS may provide a more accurate prognostication, mostly among ER/PR+ cancers and with PPS 1A patients. Patients restaged to higher PPS stages showed significant differential survival even within the same PPS. Also, only limited improvement was observed for ER–PR– cancers. Caution needs to be exercised in using PPS for patient prognostication, as in some cases the outcome can be variable with the same PPS.     

Microarray-based measurement of microRNA-449c-5p levels in hepatocellular carcinoma and bioinformatic analysis of potential signaling pathways

The clinical role and potential molecular mechanisms of microRNA-449c-5p (miR-449c-5p) in hepatocellular carcinoma (HCC) tissues remains unclear. Combining multiple bioinformatic tools, we studied the miR-449c-5p expression levels in HCC tissues and explored possible target genes and related signaling pathways. First, miR-449c-5p expression data from microarrays provided by publicly available sources were mined and analyzed using various meta-analysis methods. Next, genes that were downregulated after miR-449c-5p mimic transfection into HCC cells were identified, and in silico methods were used to predict potential target genes. Several bioinformatic assessments were also performed to evaluate the possible signaling pathways of miR-449c-5p in HCC. Five microarrays were included in the current study, including GSE98269, GSE64632, GSE74618, GSE40744 and GSE57555. The standard mean difference was 0.44 (0.07–0.80), and the area under the curve was 0.68 (0.63–0.72), as assessed by meta-analyses, which consistently indicated the upregulation of miR-449c-5p in HCC tissues. A total of 2244 genes were downregulated after miR-449c-5p mimic transfection into an HCC cell line, while 5217 target genes were predicted by in silico methods. The overlap of these two gene pools led to a final group of 428 potential target genes of miR-449c-5p. These 428 potential target genes were primarily enriched in the homologous recombination pathway, which includes DNA Polymerase Delta 3 (POLD3). Data mining with Oncomine and the Human Protein Atlas showed a decreasing trend in POLD3 mRNA and protein levels in HCC tissue samples. This evidence suggests that miR-449c-5p could play an essential role in HCC through various pathways and that POLD3 could be a potential miR-449c-5p target. However, these in silico findings should be validated with further experiments.     

Wolf-Hirschhorn综合征4例及文献复习

目的　探讨Wolf-Hirschhorn综合征（WHS）临床特征及基因突变。方法　回顾分析2017-11-20—2018-05-26南京医科大学附属儿童医院收治的4例发育延迟及智力低下患儿的临床资料，临床拟诊为WHS。应用染色体微阵列芯片分析技术进行基因检测，并复习相关文献总结疾病特点。结果　2例男性和2例女性患儿因生后特殊面容（希腊头盔样面容）、智力低下、发育延迟、肌张力低下、癫痫，应用染色体微阵列芯片分析技术发现患儿4p16.3区域2.24～3.8 Mb的缺失，确诊WHS，给予抗癫痫及康复治疗并定期随访。结论　尽早完善染色体芯片技术检查有助于早期诊断WHS，且能判断预后。染色体微阵列芯片分析与传统细胞遗传学分析方法相比，具有高分辨和高准确度的优点，可为产前遗传学诊断提供更详细信息。