Assembly of a miRNA‐modified QCM sensor for miRNA recognition through response patterns

In this paper, a miRNA‐based quartz crystal microbalance (QCM) biosensor was fabricated and used to the rapid and effective sensing of miRNA. The specific hybridization between probe miRNA and different selected miRNAs (miR‐27a, miR‐27b, and Let‐7a) cause a different interaction mode, thus display different frequency change and response patterns in the QCM sensor, which were used to detect miR‐27a and miR‐27b. The selective sensing of miR‐27a in mixed miRNA solution was also achieved. This miRNA‐based QCM biosensor has the advantages of real‐time, label‐free, and short cycle detection.     

Identification of CEACAM5 as a Biomarker for Prewarning and Prognosis in Gastric Cancer

MGd1, a monoclonal antibody raised against gastric cancer cells, possesses a high degree of specificity for gastric cancer (GC). Here we identified that the antigen of MGd1 is CEACAM5, and used MGd1 to investigate the expression of CEACAM5 in non-GC and GC tissues (N=643), as a biomarker for prewarning and prognosis. The expression of CEACAM5 was detected by immunohistochemistry in numerous tissues; its clinicopathological correlation was statistically analyzed. CEACAM5 expression was increased progressively from normal gastric mucosa to chronic atrophic gastritis, intestinal metaplasia, dysplasia and finally to GC (p<0.05). In gastric precancerous lesions (intestinal metaplasia and dysplasia), CEACAM5-positive patients had a higher risk of developing GC as compared with CEACAM5-negative patients (OR = 12.68, p<0.001). Besides, CEACAM5 was found positively correlated with invasion depth of gastric adenocarcinoma (p<0.001). In survival analysis, CEACAM5 was demonstrated to be an independent prognostic predictor for patients with GC of clinical stage IIIA/IV (p=0.033). Our results demonstrate that CEACAM5 is a promising biomarker for GC prewarning and prognostic evaluation.     

CD133 Is a Useful Surrogate Marker for Predicting Chemosensitivity to Neoadjuvant Chemotherapy in Breast Cancer

Background

Neoadjuvant chemotherapy (NAC) is a standard care regimen for patients with breast cancer. However, the pathologic complete response (pCR) rate remains at 30%. We hypothesized that a cancer stem cell marker may identify NAC-resistant patients, and evaluated CD133 and ALDH1 as a potential surrogate marker for breast cancer. The aim of this study was to find a surrogate maker to predict chemosensitivity of NAC for breast cancer.

Methodology/Findings

A total of 102 patients with breast cancer were treated with NAC consisting of epirubicin followed by paclitaxel. Core needle biopsy (CNB) specimens and resected tumors were obtained from all patients before and after NAC, respectively. Chemosensitivity and prognostic potential of CD133 or ALDH1 expression was evaluated by immunohistochemistry. Clinical CR (cCR) and pCR rates were 18% (18/102) and 29% (30/102), respectively. Forty-seven (46%) patients had CD133-positive tumors before NAC, and CD133 expression was significantly associated with a low pCR rate (p = 0.035) and clinical non-responders. Multivariate analysis revealed that CD133 expression was significantly (p = 0.03) related to pCR. Recurrence was more frequent in patients with CD133-positive tumors (21/47, 45%) than that in patients with CD133-negative tumors (7/55, 13%). The number of patients with CD133-positive tumors (62%) after NAC was higher than that (46%) before NAC. Furthermore, most patients with CD133-positive tumors before NAC maintained the same status after NAC.

Conclusion/Significance

CD133 before NAC might be a useful marker for predicting the effectiveness of NAC and recurrence of breast cancer after NAC.     

肝癌预后miRNA风险评分模型的鉴定和分析

肝细胞癌(hepatocellular carcinoma,HCC)是世界上高发病率和高死亡率的恶性肿瘤之一.研究目的是寻找HCC相关的mi RNA预后生物学标志物,预测HCC患者的风险程度和生存时间,为他们提供有效的预后信息.使用4种方法从TCGA中识别差异表达的mi RNAs(DEMs).并用Kaplan-Meier生存曲线、单因素和多因素Cox回归分析从DEMs中筛选肝癌预后相关的mi RNA.最终4个HCC的预后mi RNA生物学标志物(hsa-mi R-132-3p、hsa-mi R-139-5p、hsa-mi R-3677-3p、hsa-mi R-500a-3p)被筛选出来组合成一个风险评分模型.目前还没有实验证据表明组合中的hsa-mir-3677-3p与HCC相关,是本研究新发现的mi RNA.生存曲线、ROC曲线、卡方检验等多种生物信息学方法的评价结果均表明,该模型计算出的风险分值能有效预测患者的风险程度(P<0.000,风险比=2.551,95%置信区间=1.751-3.717).低风险组HCC患者1-5年生存率比高风险组高20%-30%.通过与临床数据分析发现,组合的生物学标志物较其他临床指标相比具有更好的预后效果,也可以作为独立的预后因子.最后,预测了4种mi RNA的靶基因,包括AGO2、FOXO1、ROCK2、RAP1B、CYLD等,并在细胞增殖、迁移、凋亡、免疫应答等生物学过程中富集.     

miR27a对结肠癌细胞增殖和侵袭能力的影响

目的:检测mi R196a、mi R146a、mi R27a和mi R200a在结肠癌患者中的表达情况,研究差异mi RNA对结肠癌细胞功能的影响。方法:用PCR检测mi R196a、mi R146a、mi R27a和mi R200a在结肠癌患者癌组织中的表达情况;用转染技术高表达和低表达mi R27a后,检测结肠癌细胞的增殖能力和侵袭能力。结果:结肠癌组mi R27a的表达水平与正常组和大肠炎组相比显著增加(P0.05);mi R27a mimics转染组结肠癌细胞的增殖速度和侵袭能力显著增高(P0.05),且mi R27a inhibitors转染组结肠癌细胞的增殖速度和侵袭能力明显降低,差异具有统计学意义(P0.05)。结论:结肠癌患者mi R27a的表达水平显著增高,且mi R27a能增强结肠癌细胞的增殖能力和侵袭能力。     

循环miRNA 表达谱在肺癌早期诊断、判断预后和指导化疗中的应用

MicroRNA(miRNA)是一种高保守,长度大概21-23个核苷酸,非蛋白编码RNA,起着调节基因表达的作用。近年来有关miRNA与肺癌的关系已经得到证实,并且成为当前研究的热点。miRNA能整体调节基因表达,这使得miRNA表达谱在作为生物信号方面比蛋白编码基因更具有提示作用。最近发现miRNA以被保护的状态存在于循环血液中,这使得miRNA表达的发现具有非侵袭性、重现性以及易检测性。研究显示血浆miRNA表达谱可作为肺癌生物信号分子,在肺癌早期诊断、判断预后和指导化疗药物应用等方面具有重要作用。本文将对血浆miRNA与肺癌的研究进展,以及在肺癌早期诊断、判断预后和指导化疗药物应用等方面作一综述。     

Caveolin-1 is a Modulator of Fibroblast Activation and a Potential Biomarker for Gastric Cancer

Stromal fibroblasts play an important role in chronic cancer-related inflammation and the development as well as progression of malignant diseases. However, the difference and relationship between inflammation-associated fibroblasts (IAFs) and cancer-associated fibroblasts (CAFs) are poorly understood. In this study, gastric cancer-associated fibroblasts (GCAFs) and their corresponding inflammation-associated fibroblasts (GIAFs) were isolated from gastric cancer (GC) with chronic gastritis and cultured in vitro. These activated fibroblasts exhibited distinct secretion and tumor-promoting behaviors in vitro. Using proteomics and bioinformatics techniques, caveolin-1 (Cav-1) was identified as a major network-centric protein of a sub-network consisting of 121 differentially expressed proteins between GIAFs and GCAFs. Furthermore, immunohistochemistry in a GC cohort showed significant difference in Cav-1 expression score between GIAFs and GCAFs and among patients with different grades of chronic gastritis. Moreover, silencing of Cav-1 in GIAFs and GCAFs using small interfering RNA increased the production of pro-inflammatory and tumor-enhancing cytokines and chemokines in conditioned mediums that elevated cell proliferation and migration when added to GC cell lines AGS and MKN45 in vitro. In addition, Cav-1 status in GIAFs and GCAFs independently predicted the prognosis of GC. Our findings indicate that Cav-1 loss contributes to the distinct activation statuses of fibroblasts in GC microenvironment and gastritis mucosa, and Cav-1 expression in both GCAFs and GIAFs may serve as a potential biomarker for GC progression.     

IL-27 和PCT 联合检测在不同分型的脓毒症危重患者中的诊断价值

目的：探讨白细胞介素-27（Interleukin 27,IL-27）对成人全身炎症反应综合征（systemic inflammatory response syndrome, SIRS）和脓毒症的诊断价值。方法：214 例SIRS患者按入院诊断结果及感染源不同分为非脓毒症组（n＝80）、肺源性脓毒症组（n＝ 73）和非肺源性脓毒症组（n＝ 61）。采用酶联免疫吸附试验（ELISA）检测各组患者血清IL-27 和降钙素原（PCT）水平;绘制受试者 工作特征曲线（ROC）,判断各指标的诊断价值,分析各生物标志物的性能,判断潜在的预测变量。结果：肺源性脓毒症患者体温符 合SIRS 标准的比例为65.8%,明显高于非脓毒症患者（45.0%）及非肺源性脓毒症患者（45.9%）（P ＜ 0.05）;非肺源性脓毒症患者白 细胞数符合SIRS标准的比例为68.9%,明显高于非脓毒症患者42.5%,（P ＜ 0.05）。确诊脓毒症后的患者血清IL-27 的AUC 为 0.655,PCT的AUC 为0.649。根据不同感染源进一步分析,肺源性和非肺源性脓毒症患者血清IL-27 水平明显高于非脓毒症患 者,肺源性和非肺源性脓毒症患者PCT 水平明显高于非脓毒症患者（P＜0.01）。ROC曲线分析发现,肺源性和非肺源性脓毒症患 者血清IL-27 的AUC分别为0.657 和0.652,肺源性和非肺源性脓毒症患者PCT 的AUC 为0.667 和0.629。分别联合检测三组患 者的血清IL-27 和PCT值,肺源性脓毒症患者的AUC为0.728,非肺源性脓毒症患者的AUC 为0.703。对肺源性脓毒症患者与非 肺源性脓毒症患者诊断的准确性均有所提升。结论：肺源性和非肺源性脓毒症患者较非脓毒症患者更加符合SIRS 标准。IL-27 作 为脓毒症诊断的生物标志物,对病情变化的反应不敏感,而IL-27 和PCT 结合可以使诊断的准确性提高。     

PCNA、P27蛋白在老年胃癌中的表达及其临床意义

目的：探讨增殖细胞核抗原（PCNA）和P27蛋白（P27）在老年胃癌中表达及其临床意义。方法：采用免疫组织化学SP法检测58例老年胃癌组织中PCNA、P27蛋白表达情况,分析它们与老年胃癌临床生物学行为的关系。结果：PCNA在老年胃癌组中的阳性表达率明显高于对照组的阳性表达率,P27在老年胃癌组中的阳性表达率明显低于对照组的阳性表达率。PCNA蛋白阳性表达与老年胃癌是否有浆膜浸润、组织分化程度、淋巴结转移以及TNM分期密切相关（P〈0.01）,而P27蛋白阳性表达与老年胃癌类型、是否有浆膜浸润、组织分化程度、淋巴结转移以及TNM分期密切相关（P〈0.01）。老年胃癌组织中PCNA、P27的表达呈负相关（r=-0.536,P〈0.05）。结论：P27表达下调、PCNA表达增强在老年胃癌的发生、发展过程中具有重要的作用,且两者具有相互作用。     

硫氧还蛋白还原酶：一种新型有效的胃癌化疗疗效监测标志物

胃癌是国际上发病率较高的肿瘤,寻找新型有效的肿瘤标志物用于胃癌的诊断和疗效评估,对于胃癌的临床治疗有较大的帮助。本研究选取经临床确诊的155例胃癌患者为研究对象,探究血浆硫氧还蛋白还原酶（thioredoxin reductase, TR）在胃癌化疗后临床获益人群与未获益人群中检测水平的差异,以及其与治疗效果的关系。结果显示,未获益组TR阳性率为82.67%,显著高于获益组TR阳性率（48.94%）。受试者操作特征曲线（receiver operating characteristic curve,ROC）分析显示,未获益组人群与获益组人群的TR活性诊断阈值为8.95 U/mL,ROC曲线下面积（area under the curve,AUC）为0.8423,明显优于常规胃癌标志物癌胚抗原（carcinoembryonic antigen,CEA）,肿瘤特异性抗原（cancer antigen,CA）19-9和CA72-4。同时,TR与常规胃癌标志物的联合检测提高阳性检出率至98.49%。TR被认为是一种新型的胃癌临床化疗疗效监测标志物,与临床疗效评价具有较高的一致性。     

硫氧还蛋白还原酶：一种新型有效的胃癌化疗疗效监测标志物

胃癌是国际上发病率较高的肿瘤,寻找新型有效的肿瘤标志物用于胃癌的诊断和疗效评估,对于胃癌的临床治疗有较大的帮助。本研究选取经临床确诊的155例胃癌患者为研究对象,探究血浆硫氧还蛋白还原酶（thioredoxin reductase, TR）在胃癌化疗后临床获益人群与未获益人群中检测水平的差异,以及其与治疗效果的关系。结果显示,未获益组TR阳性率为82.67%,显著高于获益组TR阳性率（48.94%）。受试者操作特征曲线（receiver operating characteristic curve,ROC）分析显示,未获益组人群与获益组人群的TR活性诊断阈值为8.95 U/mL,ROC曲线下面积（area under the curve,AUC）为0.8423,明显优于常规胃癌标志物癌胚抗原（carcinoembryonic antigen,CEA）,肿瘤特异性抗原（cancer antigen,CA）19-9和CA72-4。同时,TR与常规胃癌标志物的联合检测提高阳性检出率至98.49%。TR被认为是一种新型的胃癌临床化疗疗效监测标志物,与临床疗效评价具有较高的一致性。     

骨碎补对骨性关节炎患者骨关节滑膜组织中miR-27a、miR-146a表达的影响

目的:探讨骨碎补对骨性关节炎患者骨关节滑膜组织中mi R-27a、mi R-146a表达的影响。方法:选取我院收治的骨性关节炎患者46例,随机分为两组,每组各23例。对照组予抗骨增生治疗,实验组在对照组基础上加以骨碎补治疗。观察和比较两组患者的临床疗效、炎性因子水平的变化以及骨关节滑膜组织中mi R-27a、mi R-146a的表达情况。结果:1治疗后,两组患者的临床症状均有所改善,且实验组疗效高于对照组,差异有统计学意义(P0.05)。2治疗后,两组患者的IL-1β、PGE2水平均较治疗前显著降低,且实验组降低较对照组更明显,差异均有统计学意义(P0.05)。3治疗后,两组患者的骨关节滑膜组织中的mi R-27a、mi R-146a表达均降低,且与对照组比较,实验组较低,差异有统计学意义(P0.05)。结论:骨碎补能够降低骨性关节炎患者骨关节滑膜组织中mi R-27a、mi R-146a的表达,降低IL-1β、PGE2的水平,提高临床疗效。     

培美曲塞联合长春瑞滨治疗复发转移性宫颈癌患者对疾病控制及预后随访研究

摘要 目的：探讨培美曲塞联合长春瑞滨治疗复发转移性宫颈癌对疾病控制及预后的影响。方法：选择2018年5月~2019年6月收治的100例复发转移性宫颈癌患者,按照随机数表法分观察组和对照组,每组50例。对照组采用紫杉醇联合顺铂化疗,观察组则采用培美曲塞联合长春瑞滨治疗。比较2组疾病控制效果,记录患者化疗前后Karnofsky评分变化,统计2组患者毒副反应及生存率、生存期。结果：观察组的疾病控制率为84.00%,明显高于对照组的62.00%,差异显著（P<0.05）;治疗后,2组患者的Karnofsky评分均升高,且观察组[（86.25±3.02）分]明显高于对照组[（81.15±3.11）分],差异显著（P<0.05）; 2组患者胃肠道反应、骨髓抑制、血小板计数减少、恶心呕吐等毒副反应发生率比较,无明显差异（P>0.05）;观察组患者的1年生存率70.00%、2年生存率58.00%、3年生存率26.00%均高于对照组的50.00%、38.00%、10.00%,观察组的生存期[（18.12±3.21）月]长于对照组[（14.18±2.81）月],差异显著（P<0.05）。结论：培美曲塞联合长春瑞滨治疗复发转移性宫颈癌效果显著,可有效提高患者的生存质量及生存率,延长生存期,毒副反应低,患者耐受性好,可在临床推广运用。     

Loss of TRAIL-Receptors Is a Recurrent Feature in Pancreatic Cancer and Determines the Prognosis of Patients with No Nodal Metastasis after Surgery

Introduction

Agonistic antibodies targeting TRAIL-receptors 1 and 2 (TRAIL-R1 and TRAIL-R2) are being developed as a novel therapeutic approach in cancer therapy including pancreatic cancer. However, the cellular distribution of these receptors in primary pancreatic cancer samples has not been sufficiently investigated and no study has yet addressed the issue of their prognostic significance in this tumor entity.

Aims and Methods

Applying tissue microarray (TMA) analysis, we performed an immunohistochemical assessment of TRAIL-receptors in surgical samples from 84 consecutive patients affected by pancreatic adenocarcinoma and in 26 additional selected specimens from patients with no lymph nodes metastasis at the time of surgery. The prognostic significance of membrane staining and staining intensity for TRAIL-receptors was evaluated.

Results

The fraction of pancreatic cancer samples with positive membrane staining for TRAIL-R1 and TRAIL-R2 was lower than that of cells from surrounding non-tumor tissues (TRAIL-R1: p<0.001, TRAIL-R2: p = 0.006). In addition, subgroup analyses showed that loss of membrane staining for TRAIL-R2 was associated with poorer prognosis in patients without nodal metastases (multivariate Cox regression analysis, Hazard Ratio: 0.44 [95% confidence interval: 0.22−0.87]; p = 0.019). In contrast, analysis of decoy receptors TRAIL-R3 and -R4 in tumor samples showed an exclusively cytoplasmatic staining pattern and no prognostic relevance.

Conclusion

This is a first report on the prognostic significance of TRAIL-receptors expression in pancreatic cancer showing that TRAIL-R2 might represent a prognostic marker for patients with early stage disease. In addition, our data suggest that loss of membrane-bound TRAIL-receptors could represent a molecular mechanism for therapeutic failure upon administration of TRAIL-receptors-targeting antibodies in pancreatic cancer. This hypothesis should be evaluated in future clinical trials.     

Rab27a is required for regulated secretion in cytotoxic T lymphocytes

Rab27a activity is affected in several mouse models of human disease including Griscelli (ashen mice) and Hermansky-Pudlak (gunmetal mice) syndromes. A loss of function mutation occurs in the Rab27a gene in ashen (ash), whereas in gunmetal (gm) Rab27a dysfunction is secondary to a mutation in the alpha subunit of Rab geranylgeranyl transferase, an enzyme required for prenylation and activation of Rabs. We show here that Rab27a is normally expressed in cytotoxic T lymphocytes (CTLs), but absent in ashen homozygotes (ash/ash). Cytotoxicity and secretion assays show that ash/ash CTLs are unable to kill target cells or to secrete granzyme A and hexosaminidase. By immunofluorescence and electron microscopy, we show polarization but no membrane docking of ash/ash lytic granules at the immunological synapse. In gunmetal CTLs, we show underprenylation and redistribution of Rab27a to the cytosol, implying reduced activity. Gunmetal CTLs show a reduced ability to kill target cells but retain the ability to secrete hexosaminidase and granzyme A. However, only some of the granules polarize to the immunological synapse, and many remain dispersed around the periphery of the CTLs. These results demonstrate that Rab27a is required in a final secretory step and that other Rab proteins also affected in gunmetal are likely to be involved in polarization of the granules to the immunological synapse.