黄磷染毒对小鼠肝脏HSP70表达及NO的影响

[目的]观察黄磷染毒对小鼠HSP70表达及NO的影响，以探讨黄磷的毒作用机制，寻求黄磷中毒早期健康效应的生物标志物。[方法]通过对小鼠黄磷急性和亚急性毒理实验，应用生物化学和分子生物学的技术，选取176只健康雄性昆明种小鼠，体重18—22g，检测黄磷染毒对HSP70、一氧化氮(NO)、一氧化氮合酶(NOS)等指标的影响。染毒时，急性剂量．效应关系研究取小鼠32只，随机分成4组，低、中、高剂量组的染磷量分别为3、6、9mg／kg体重、对照组给予等量的花生油，腹腔注射染毒48h。时间--效应关系研究取小鼠64只，随机分成8组，实验组以9mg／kg体重的剂量一次性腹腔注射染毒，分别于染毒O、12、24、48h后断头处死小鼠，各组配以相应的对照组。亚急性毒理实验取小鼠80只，随机分成以下10组：O、1、2、3、4周时问组，以1．5mg／kg体重的剂量隔日腹腔注射染毒，每组配以相应的对照组。[结果]随染磷剂量的增高和染磷时间的延长，小鼠肝匀浆HSP70的表达也逐渐升高，急性染磷的HSP70表达升高比亚急性染磷明显；小鼠肝匀浆NOS活性和NO含量逐渐升高，并呈现良好的剂量--反应关系和时间--效应关系。[结论]在黄磷染毒过程中，HSP70对细胞起着很重要的保护作用。提示NOS和NO有可能作为黄磷中毒早期健康效应的生物标志物。     

生物标记物在宫颈癌鉴别诊断中的应用

病理学诊断在宫颈病变的诊断中发挥重要作用,用免疫组化法检测生物标记物,并与目前的组织学标准联合应用,可大大提高妇产科疾病诊断的一致性和可重复性。     

Pivoting Novel Exosome-Based Technologies for the Detection of SARS-CoV-2

The National Institutes of Health (NIH) launched the Rapid Acceleration of Diagnostics (RADx) initiative to meet the needs for COVID-19 diagnostic and surveillance testing, and to speed its innovation in the development, commercialization, and implementation of new technologies and approaches. The RADx Radical (RADx-Rad) initiative is one component of the NIH RADx program which focuses on the development of new or non-traditional applications of existing approaches, to enhance their usability, accessibility, and/or accuracy for the detection of SARS-CoV-2. Exosomes are a subpopulation of extracellular vesicles (EVs) 30–140 nm in size, that are critical in cell-to-cell communication. The SARS-CoV-2 virus has similar physical and molecular properties as exosomes. Therefore, the novel tools and technologies that are currently in development for the isolation and detection of exosomes, may prove to be invaluable in screening for SARS-CoV-2 viral infection. Here, we describe how novel exosome-based technologies are being pivoted for the detection of SARS-CoV-2 and/or the diagnosis of COVID-19. Considerations for these technologies as they move toward clinical validation and commercially viable diagnostics is discussed along with their future potential. Ultimately, the technologies in development under the NIH RADx-Rad exosome-based non-traditional technologies toward multi-parametric and integrated approaches for SARS-CoV-2 program represent a significant advancement in diagnostic technology, and, due to a broad focus on the biophysical and biochemical properties of nanoparticles, the technologies have the potential to be further pivoted as tools for future infectious agents.     

Identification of Potential mRNA Biomarkers in Milk Small Extracellular Vesicles of Enzootic Bovine Leukosis Cattle

Enzootic bovine leukosis (EBL) is a disease caused by bovine leukemia virus (BLV); only a small percentage of BLV-infected cattle develop EBL and present with B-cell lymphosarcoma. There is no vaccine against BLV, treatment for EBL, or method for predicting the possibility of EBL onset, thus making EBL control difficult. Herein, to explore biomarkers for EBL in milk, we examined the mRNA profiles of small extracellular vesicles (sEVs) in milk from four BLV-uninfected and four EBL cattle by microarray analysis. It was revealed that 14 mRNAs were encapsulated in significantly higher quantities, and these mRNAs were therefore selected as biomarker candidates. Primers for these mRNAs were designed, and nine primer sets were available for quantitative real-time PCR. Nine mRNAs were evaluated for their availability as biomarkers for EBL using sEVs from newly-collected milk of 7 uninfected and 10 EBL cattle. The quantities of eight mRNAs (TMEM156, SRGN, CXCL8, DEFB4A, FABP5, LAPTM5, LGALS1, and VIM) were significantly higher in milk sEVs of EBL cattle than in those of uninfected cattle. Therefore, our findings indicate that these eight mRNAs in milk sEVs can be used as potential EBL biomarkers with combination use, although single mRNA use is not enough. Consequently, cattle at risk of EBL onset can be identified by monitoring the fluctuation in quantities of these mRNAs in milk before they develop EBL.     

p16基因甲基化与胃黏膜异型增生恶性转化的关系

目的 山东省临朐县是我国北方胃癌高发区之一,人群中胃黏膜异型增生(dysplasia,DYS)病变所占比例较高.DYS是胃黏膜癌前病变发展的高级阶段,具有显著的恶性转化潜能.本研究探讨p16基因甲基化与DYS病变恶性转化的关系,验证其作为胃癌预警标志物的应用价值.方法 以胃癌高发现场长期胃镜随访队列为基础,对101例来自山东省临朐县胃癌高发区且随访至2015-12-31的DYS病例,采用Methylight方法定量检测胃黏膜组织p16基因甲基化水平,评价其与DYS进展为胃癌风险的关系.结果 在进展为胃癌组,p16基因甲基化百分比中位数(四分位数)为1.30％(0.13％～1.87％),显著高于非进展组的0.67％(0～1.15％),P=0.047.与p16基因低甲基化者相比,高甲基化水平的DYS患者进展为胃癌的风险显著增加,OR-3.67,95％CI为1.31～10.34.进一步分析发现幽门螺杆菌(H.pylori)感染阳性者p16基因甲基化百分比中位数(0.98％)明显高于阴性者(0.01％),P＜0.001;分层分析发现,p16基因高甲基化同时H.pylori感染阳性者进展为胃癌的风险进一步增加,OR=5.14,95％CI为1.57～16.82.通过分析p16基因甲基化水平与DYS病例进展为胃癌的时间关系,发现随着胃癌诊断时间的临近,p16基因甲基化水平有缓慢升高的趋势,但差异未见统计学意义.结论 胃黏膜组织p16基因甲基化水平升高可作为DYS患者发生恶性转化的潜在预警标志物.     

Identification and verification of Hsp90-beta as a potential serum biomarker for lung cancer

Background: Hsp90-beta was investigated as prognostic factor because of its apparent association with tumorigenesis. The aim of this study was to investigate the expression of Hsp90-beta in lung cancer patients, to analyze the relationship with respect to the clinicopathological features and to assess whether Hsp90-beta as a potential serum marker for lung cancer. Methods: Expression of Hsp90-beta was examined using immunohistochemistry, in-situ hybridization, western blot and enzyme-linked immunosorbent assay. Sensitivities and specificities for Hsp90-beta serum test were determined using receiver operator characteristic curve and cutoff was defined based on 95% and 85% sensitivities. Results: Lung cancer tissues exhibited higher expression of Hsp90-beta than the normal tissues (P < 0.05) and the serum Hsp90-beta of lung cancer patients also exhibited higher level than control groups (P < 0.05). Moreover, increased serum Hsp90-beta was significantly associated with the pathological grade and clinical stage of lung cancer patients (P < 0.05). Using receiver operator characteristic curve analysis, the cutoffs for distinguishing lung cancer from normal and benign groups were 1.155 and 1.158 ng/ml respectively. The sensitivities of Hsp90-beta for distinguishing lung cancer from normal and benign groups were 98.77% and 95.9%, and specificities were 88.33% and 72.7%. Conclusion: Up-regulation of serum Hsp90-beta was associated with pathological grade and clinical stage of lung cancer patients, which indicated that it could be considered molecular biomarker for diagnosis and prognosis of lung cancer.     

血清中microRNA-210表达水平对胃癌的诊断价值

目的:microRNA-210(miRNA-210)在胃癌组织中表达上调,血液中也能检测到miRNA-210的表达,本研究旨在探索血清中miRNA-210的表达水平对胃癌的诊断价值.方法:选取100例胃癌患者和100例健康成人,血清中miR-210的表达检测采用实时定量PCR法.评估循环血中miR-210用于胃癌诊断的敏感性、特异性,分析血清中miR-210对不同分期胃癌的诊断价值.结果:胃癌患者血清中miR-210较健康对照组显著升高,血清中miR-210对胃癌诊断的敏感性和特异性分别为85.6％和82.4％,血清miR-210对胃癌Ⅰ到Ⅳ期的阳性预测值分别为89.95％,90.31％,90.45％和91.21％.结论:血清miR-210可作为胃癌诊断的生物标志物.     

乳腺癌临床病理指标以及分子分型对TEC新辅助化疗病理完全缓解的预测价值

目的  本研究旨在探讨乳腺癌临床病理指标以及乳腺癌分子分型对多西他赛联合表柔比星、环磷酰胺（TEC）的 新辅助化疗后病理完全缓解率（pathological complete response pCR）的预测价值。方法   对 214例经4周期TEC新辅助化疗的乳腺癌患者的临床病理资料进行回顾性分析;免疫组织化学检测经核心针穿刺 的癌组织雌激素受体（ER）、孕激素受体（PR）、人类表皮生长因子受体-2（HER2）、Ki67、p53表达情况,原位基 因免疫荧光杂交（FISH）检测HER2有无过表达;根据ER、PR、HER2、Ki67的表达情况将乳腺癌分为4种分子分型： LuminalA、 LuminalB、HER2过表达型和三阴性乳腺癌。分析不同的临床病理指标、不同分子分型与pCR的相关性。结果  4周期TEC新辅助化疗后pCR率为14.0%（30/214）;单因素分析：ER、PR、Ki67、乳腺癌分子分型与pCR均具有显 著相关性（P＜0.05）;乳腺癌分子分型各组间显示pCR率不同：LuminalA＜LuminalB＜HER2过表达型＜三阴性乳腺癌 ;多因素分析：与pCR具有显著相关性的分类变量为ER（OR=0.311,95%CI：0.136～0.712;P=0.006）和Ki67 （OR=2.788,95%CI：1.061～7.327;P=0.038）。结论  ER、PR、Ki67以及乳腺癌分子分型可能是TEC新辅助化疗后乳腺癌pCR的预测指标。