活细胞样本的液基制片及染色

病理学中的活细胞样本是指将肿瘤细胞以较低密度接种于液体或半固体的培养液中形成悬浮培养的细胞系,对于已培养的活细胞需要转移至玻片上进行染色后镜下细胞形态学观察及细胞性质的鉴定。本实验将活细胞悬液转移至液基固定瓶中进行薄层制片,经HE染色和免疫细胞化学染色检测取得良好效果,现介绍如下。

微流控芯片上细胞样本低温保存的研究进展

随着分子、细胞以及微组织、器官模型等逐渐转移到微流控芯片上,微流控芯片的平台性价值在生物医学领域日益凸显。发展芯片上细胞样本的低温保存技术不仅是实现上述价值的重要条件,也是对传统细胞样本低温保存的有益创新。对微流控芯片上细胞样本的低温保存技术进行了综述,包括慢速冷冻保存及其升降温过程的控制、超快速降温玻璃化保存的理论分析与验证,并在此基础上进一步介绍了芯片上细胞样本的常温保存,最后对该领域进展中存在的动力与需要解决的问题作了简要分析与展望,以期为相关研究提供参考。

肺癌组织样本和血液样本EGFR基因检测对比研究

目的:研究肺腺癌患者不同样本EGFR基因检测结果的异同,以更精准地反映患者体内EGFR突变情况。方法:收集2016年7月—2017年12月收治的Ⅲ~Ⅳ期经病理确诊为肺腺癌患者175例,175例均有血浆样本,其中119例同时有活检组织样本,12例同时有胸水沉渣细胞样本。采用突变扩增系统(ARMS)-聚合酶链式反应(PCR)方法对收集到的活检组织样本、胸水沉渣细胞样本和血液样本进行EGFR基因检测,分析不同类型样本间结果的差异,以及与患者临床病理指标的关系。结果:联合检测的EGFR基因突变总阳性率较单纯血液样本阳性率提高了8.6%,较单纯组织样本提高了3.1%。EGFR突变在不同的性别(χ^(2)=3.451,P=0.045)、肿瘤直径(χ^(2)=8.911,P=0.002)、T分期(χ^(2)=17.567,P=0.000)、淋巴结转移(χ^(2)=18.511,P=0.000)、转移站数(χ^(2)=3.341,P=0.047)和远处转移(χ^(2)=10.874,P=0.001)组间的差异有统计学意义。将血液和组织两组样本EGFR突变率进行比较,差异有统计学意义(P=0.000)。在患者临床病理指标与生存时间的分析中,肿瘤直径[95%CI(0.151,0.725),P=0.006]、淋巴结转移[95%CI(0.180,0.865),P=0.020]、远处转移[95%CI(0.097,0.612),P=0.003]均是影响患者无进展生存期的重要因素。结论:肺癌患者血液样本、活检组织样本、胸水沉渣细胞样本在EGFR检测过程中的联合应用能够获得更高的检出率,可为临床更多地筛选出适合EGFR-酪氨酸激酶抑制剂(TKIs)治疗的人群。对于女性患者、肿瘤直径≥3 cm、T4期、淋巴结转移且超过3站、远处转移患者检测到血液EGFR基因突变的几率更高。其中肿瘤直径≥3 cm、淋巴结转移、远处转移患者的无进展生存期更短。

一种细胞固定和密度估算及塑形一体化装置设计

目的 开发一种用于脱落细胞样本固定、细胞密度估算和细胞团块制作的装置,并验证其处理临床脱落细胞样本的效果。方法 采用Auto CAD软件和三维立体(3D)打印技术,设计制作本装置中的离心管,然后以黑白打印机打印密度标签。采集脱落细胞样本,通过观察密度标签不同灰度区域中的黑色线条是否可被清晰观察,评估细胞密度;随后将样本装入该离心管,离心获得细胞团块并制作石蜡细胞芯片,切片后进行苏木素-伊红(HE)染色和免疫细胞化学(ICC)染色,对染色结果进行观察。结果 该研究以经病理检测后剩余的12例胸/腹水为样本,采用该装置处理,获得直径4 mm、高1~10 mm不等的圆柱形细胞团块。HE染色结果显示,细胞分布均匀无重叠且形态完整,胞膜、质、核结构清晰;ICC染色结果显示,广谱细胞角蛋白(CK pan)、细胞核相关抗原(Ki67)阳性信号强、定位准确、背景清晰。结论 采用该装置制作脱落细胞蜡块,HE染色和ICC染色效果良好,可获得直径一致的圆柱形细胞团块,有利于建立标准化的脱落细胞蜡块制作流程,具有较高的临床应用价值。

成都市武侯区妇女高危型人乳头瘤病毒的流行病学研究

目的探讨四川省成都市武侯区妇女高危型人乳头瘤病毒(HPV)的流行病学特征。方法选取四川省妇幼保健院行体检的14 000名妇女,根据年龄分为A组(≤35岁)、B组(> 35岁～≤45岁)、C组(> 45岁～≤55岁)、D组(> 55岁～≤65岁)。检测并比较各组宫颈脱落细胞样本中高危型HPV阳性比例及病毒类型。结果本研究妇女高危型HPV总感染率为9. 14%,其中C组感染率最高(9. 94%),显著高于D组(9. 57%)、B组(8. 90%)、A组(7. 93%)感染率(P <0. 05)。基因分型结果显示,单一感染率最高的为HPV-52型,感染率为21. 41%。各高危型HPV亚型在不同年龄段组分布无显著差异(P>0. 05)。随着年龄的增加,人群中感染2种以上HPV的概率增大,D组感染3、4、5种HPV的比例最高(P <0. 05)。结论高危型HPV的感染与年龄有关系,妇女应适时进行体检筛查以降低宫颈癌的发生概率。

超越人类基因组

掌握了编码人类的所有DNA，随之而来的挑战就是怎样利用它了。也许一些最早的成果将出自称为蛋白组学(proteomics)的这一新兴领域。

Intrahepatic natural killer T cell populations are increased in human hepatic steatosis

AIM: To determine if natural killer T cell (NKT) populations are affected in nonalcoholic fatty liver disease (NAFLD). METHODS: Patients undergoing bariatric surgery underwent liver biopsy and blood sampling during surgery. The biopsy was assessed for steatosis and immunocyte infiltration. Intrahepatic lymphocytes (IHLs) were isolated from the remainder of the liver biopsy, and peripheral blood mononuclear cells (PBMCs) were isolated from the blood. Expression of surface proteins on both IHLs and PBMCs were quantified using flow cytometry. RESULTS: Twenty-seven subjects participated in thisstudy. Subjects with moderate or severe steatosis had a higher percentage of intrahepatic CD3+/CD56+ NKT cells (38.6%) than did patients with mild steatosis (24.1%, P = 0.05) or those without steatosis (21.5%, P = 0.03). Patients with moderate to severe steatosis also had a higher percentage of NKT cells in the blood (12.3%) as compared to patients with mild steatosis (2.5% P = 0.02) and those without steatosis (5.1%, P = 0.05). CONCLUSION: NKT cells are significantly increased in the liver and blood of patients with moderate to severe steatosis and support the role of NKT cells in NAFLD.

The peri-operative changes of T-lymphocyte subsets in the elder esophageal carcinoma patients

Objective:The aim of the study was to observe and compare the trend of T-lymphocyte subsets in the elder and adult esophageal carcinoma patients postoperatively. Methods: Forty-four esophageal carcinoma patients, classified as ASA physical status I-II, were divided into two groups. Twenty-two patients aged than 65 years were grouped as elder patient group, and the rest twenty-two patients, served as adult patient group, were younger than 60 years old. The jugular venous blood samples were collected before induction of anesthesia (T1) as baseline, after completion of surgery (T2), on the first, second and fifth postoperative days (T3, T4 and T5), respectively. The blood counts of CD3+, CD4+ and CD8+T-lymphocyte were measured by flow cytometer. Results: Compared with the baseline level, the blood CD3+, CD4+ and CD4/CD8 levels were significantly decreased immediately after surgery, on the first and second postoperative days in the two groups (P<0.01), which returned to baseline values on the fifth postoperative day (P>0.05). On the fifth postoperative day, CD3+, CD4+ and CD4/CD8 levels in adult group were significantly higher than those in elder group (P<0.05). Conclusion: The postoperative immune function in elder patients recovered more slowly than that in adult.

The cancer genomics and global cancer genome collaboration

All cancers arise as a result of abnormalities occurring in the DNA sequence of cancer cells, and we are now stepping into an era in which it is feasible to obtain the complete DNA sequence of large cohorts of cancer patients. The International Cancer Genome Consortium （ICGC） launched in 2007 is devoted to coordinate large-scale cancer genome studies in tumors from 50 different cancer types and/or subtypes and systematic studies of more than 25,000 cancer genomes. Several participant groups have summa- rized and published their data for various cancers. As the active members of ICGC, Chinese cancer genome investi- gators have contributed research for 13 tumor types and released some research articles about esophageal, liver, bladder, and kidney cancers. As genetic alterations in thousands of tumors have now been catalogued, the pan- cancer analysis has become ICGC at present. The ICGC the most significant role of research network will reveal the repertoire of oncogenic mutations, uncover traces of the mutagenic influences, define molecular subtypes for clinicalimplication, and enable the development of individual therapeutics for human cancers.