结直肠癌隐匿性腹膜转移风险预测模型的建立和验证

目的利用血液学参数和临床病理特征建立用于术前预测结直肠癌(colorectal cancer,CRC)隐匿性腹膜转移的风险预测模型并进行验证。方法回顾性地收集2015年7月至2021年7月在武汉大学中南医院收治并行手术治疗的710例CRC患者的资料,基于手术时间按3∶1比例分为训练集和内部验证集,并收集2019年7月到2021年7月在长江大学荆州市中心医院的193例CRC患者作为独立的外部验证集。通过套索-逻辑回归算法(the least absolute shrinkage and selective operator-Logistic,LASSO-Logistic)筛选出的预测因子建立风险预测模型,建立术前预测CRC隐匿性腹膜转移的诺模图。从区分度、校准能力和临床净收益等方面验证诺模图的性能和临床效益。结果从血液学参数和临床病理特征中筛选出6个预测因子:肿瘤病理类型、浸润深度、影像学腹水、糖类抗原125(carbohydrate antigen 125,CA125)、糖类抗原199(carbohydrate antigen 199,CA199)和D-二聚体。建立了结合血液学参数和临床病理特征的模型,并基于模型构建了用于术前预测CRC隐匿性腹膜转移的诺模图。诺模图在训练集、内部验证集和外部验证集的受试者工作特征曲线下面积分别为:0.956(95%CI:0.936~0.975)、0.891(95%CI:0.857~0.925)和0.901(95%CI:0.860~0.942),校准曲线和临床决策分析曲线验证了模型较好的校准能力和临床净收益。在训练集中,采用约登指数获得分组的最佳截断值为206.6,此时的灵敏度为94.3%,特异度为86.6%,Kappa值为0.753,说明模型预测结果与实际情况具有较高的真实性和一致性,模型具有较好的临床预测性能。结论结合血液学参数和临床病理特征构建了预测CRC隐匿性腹膜转移的风险预测模型,其具有良好的区分度,校准能力和临床净收益,能为CRC隐匿性腹膜转移的诊断和治疗决策提供参考。

一种喹啉类锌离子荧光探针的合成及荧光性质研究

合成和表征了一种以喹啉为荧光团的锌离子探针FP1,并研究了它的荧光性质。结果显示,FP1对Zn^(2+)具有较好的选择性和灵敏度,响应快,抗干扰能力强,它对Zn^(2+)的线性响应范围在0.2~3.8μmol/L之间,与Zn^(2+)之间的结合比为1∶1。FP1能可视化识别Zn^(2+),在不同水样中对Zn^(2+)的检测表明它在环境污染监测领域将具有潜在的应用价值。

Inhibition of Adhesion and Metastasis of HepG2 Hepatocellular Carcinoma Cells In Vitro by DNA Aptamer against Sialyl Lewis X

The sialyl Lewis X（SLe;） antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin（E-selectin）. The combination of SLe;antigen and E-selectin represents an important way for malignant tumor metastasis. In the present study, the effect of the SLe;-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated. Reverse transcription-polymerase chain reaction（RT-PCR） and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels. The SLe;expression in HepG2 cells treated with different concentrations of SLe;-binding DNA aptamer was detected by flow cytometry. Besides, the adhesion, migration, and invasion of HepG2 cells were measured by cell adhesion assay, and the Transwell migration and invasion assay. The results showed that the FUT7 expression was up-regulated at both mR NA and protein levels in HepG2 cells. SLe;-binding DNA aptamer could significantly decrease the expression of SLe;in HepG2 cells. The cell adhesion assay revealed that the SLe;-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLe;in the HepG2 cells. Additionally, SLe;-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1. The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5, 10, 20 nmol/L SLe;-binding DNA aptamer than those in the negative control group（P<0.01）. Our study demonstrated that the SLe;-binding DNA aptamer could significantly inhibit the in vitro adhesion, migration, and invasion of HepG2 cells, suggesting that the SLe;-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.

Knockdown of GRHL3 Inhibits Activities and Induces Cell Cycle Arrest and Apoptosis of Human Colorectal Cancer Cells

The Grainyhead-like 3（GRHL3） is involved in epidermal barrier formation, neural tube closure and wound repair. Previous studies have suggested that GRHL3 has been linked to many different types of cancers. However, to date, its effects on human colorectal cancer（CRC） has not been clarified yet. Our microarray analysis has indicated predominant GRHL3 expression in CRC. The purpose of this study was to investigate the expression and significance of GRHL3 in CRC tumorigenesis using CRC tissues and paired paracancerous tissues, as well as using distinct CRC cell lines（HT29 and DLD1）. We observed increased GRHL3 expression at both m RNA and protein levels in CRC tissues and CRC cell lines using quantitative real-time polymerase chain reaction（q RT-PCR） and Western blotting. Moreover, silencing GRHL3 with si RNA could suppress CRC cell proliferation, viability and migration in vitro. We also found that knockdown of GRHL3 could promote cell cycle arrest at G0/G1 phase in HT29 cells and DLD1 cells, and induce cell apoptosis in HT29 cells. Together, our study revealed the down-regulation of GRHL3 in vitro could inhibit CRC cell activity and trigger cell cycle arrest at G0/G1 phase and apoptosis.

三野淋巴结清扫术在食管癌手术中的应用

食管癌是常见恶性肿瘤之一,尽管多学科治疗手段提高了食管癌的预后,但其病死率仍较高,因为食管癌淋巴结转移可首先出现在颈部至腹部,所以采取了扩大淋巴结清扫的策略;三野淋巴结清扫术（3FLD）建立于20世纪80年代的日本,定义为一种经胸食管切除术,其进行颈-胸-腹三野淋巴结清扫,目前其被全世界广泛接受.迄今为止,已经报道各种关于3FLD和二野淋巴结清扫术（2FLD）的对比试验,并且证明3FLD在预后方面优于2FLD.但是,在3FLD中出现众多术后并发症如喉返神经损伤和术后胃肠道功能紊乱.近来,食管癌术前治疗的功效得到报道,微创手术的重要性也正在得到验证.术前治疗和可行的手术方法相结合,其将在未来不断提高食管癌患者的预后.

结直肠癌患者错配修复缺陷相关预后基因的筛选

目的探索结直肠癌患者中高微卫星不稳定性(MSI-H)相关的关键预后基因。方法从癌症基因组图谱(TCGA)数据库纳入377例结直肠癌患者, 筛选MSI-H和错配修复稳定(MSS)患者的差异表达基因(阈值为错误发现率<0.05且倍数差异>2), 并对其进行富集分析(错误发现率<0.05)。利用Pearson相关分析构建基因共表达网络(R>0.4且P<0.05), 并筛选枢纽基因。通过Cox回归进一步筛选出肿瘤组织中高表达的关键预后基因(P<0.05), 采用单细胞测序分析验证关键基因表达及与MSI-H的关系。不同患者组间的基因表达差异采用t检验。结果共筛选出558个MSI-H相关基因, 811个MSS相关基因。富集分析显示MSI-H相关基因主要参与炎症及免疫应答。通过网络分析及Cox回归, 鉴定出3个MSI-H显著相关的预后关键基因, 即GJB5、TNNT1和KRT16。其在结直肠癌肿瘤组织中高表达, 且在转移性患者中对总生存期的风险比分别为1.4(P<0.05), 1.5(P<0.01)和1.7(P<0.01)。在单细胞测序数据中, 同样发现GJB5、TNNT1和KRT16在肿瘤细胞和MSI-H患者细胞中高表达。结论 GJB5、TNNT1、KRT16为MSI-H结直肠癌患者高表达的显著不良预后关键基因。