Genomic characterization of small cell carcinomas of the uterine cervix

Small cell carcinoma (SCC) of the uterine cervix is a rare and aggressive form of neuroendocrine carcinoma, which resembles small cell lung cancer (SCLC) in its histology and poor survival rate. Here, we sought to define the genetic underpinning of SCCs of the uterine cervix and compare their mutational profiles with those of human papillomavirus (HPV)‐positive head and neck squamous cell carcinomas, HPV‐positive cervical carcinomas, and SCLCs using publicly available data. Using a combination of whole‐exome and targeted massively parallel sequencing, we found that the nine uterine cervix SCCs, which were HPV18‐positive (n = 8) or HPV16‐positive (n = 1), harbored a low mutation burden, few copy number alterations, and other than TP53 in two cases no recurrently mutated genes. The majority of mutations were likely passenger missense mutations, and only few affected previously described cancer‐related genes. Using RNA‐sequencing, we identified putative viral integration sites on 18q12.3 and on 8p22 in two SCCs of the uterine cervix. The overall nonsilent mutation rate of uterine cervix SCCs was significantly lower than that of SCLCs, HPV‐driven cervical adeno‐ and squamous cell carcinomas, or HPV‐positive head and neck squamous cell carcinomas. Unlike SCLCs, which are reported to harbor almost universal TP53 and RB1 mutations and a dominant tobacco smoke‐related signature 4, uterine cervix SCCs rarely harbored mutations affecting these genes (2/9, 22% TP53; 0% RB1) and displayed a dominant aging (67%) or APOBEC mutational signature (17%), akin to HPV‐driven cancers, including cervical adeno‐ and squamous cell carcinomas and head and neck squamous cell carcinomas. Taken together, in contrast to SCLCs, which are characterized by highly recurrent TP53 and RB1 alterations, uterine cervix SCCs were positive for HPV leading to inactivation of the suppressors p53 and RB, suggesting that these SCCs are convergent phenotypes.     

Identification of HPV genotypes causing cervical precancer using tissue-based genotyping

Identification of high-risk human papillomavirus genotypes causing cervical precancer is crucial for informing HPV vaccine development and efficacy studies, and for determining which types to include in next-generation genotyping assays. Co-occurrence of hrHPV infections is common and complicates carcinogenicity assessment; accurate attribution requires tissue-based genotyping of precancers. We included all women with cervical intraepithelial neoplasia Grade 2 or worse (CIN2+) from the Biopsy Study, an observational study of 690 women enrolled between 2009 and 2012 at the University of Oklahoma. Tissue-based genotyping, including whole tissue sections (WTS) and laser-capture microdissection (LCM), was performed on all precancers with multiple hrHPV infections detected in cytology, totaling over 1,800 HPV genotyping assays. Genotype attribution was compared to hierarchical and proportional hrHPV-type attribution models. Of 276 women with CIN2+, 122 (44.2%) had multiple hrHPV genotypes in cytology. Of 114 women with genotyping data, 94 had one or more hrHPV detected in tissue. Seventy-one women (75.5%) had a single causal hrHPV genotype, while 23 women had multiple hrHPV genotypes causing CIN2+. Ten women had multiple causal infections in a single biopsy, contrary to the previous notion that each lesion is caused by a single type only. While HPV16 was the predominant causal hrHPV genotype using all approaches, the hierarchical model overattributed HPV16, whereas other causal hrHPV genotypes, particularly HPV18 and HPV35, were underattributed. Understanding true causal genotypes is important for the evaluation of vaccine efficacy, to estimate the extent of unmasking, and for type-specific risk assessment in screening and management.     

Long‐term effectiveness of HPV vaccination against HPV infection in young Japanese women: Real‐world data

In Japan, public funding for HPV vaccination began in 2010 for girls aged 13–16 years (birth cohort years 1994–1997) and women born in 1994 who turned 25 in 2019. We aimed to verify the long‐term effectiveness of the bivalent HPV vaccine in women aged 25 years. Subjects were women aged 25–26 years who underwent cervical cancer screening and HPV testing in Niigata from 2019 to 2020 (birth cohort years 1993–1994). Information on vaccination status and sexual behavior was obtained from a questionnaire and municipal records. We compared the HPV infection rates of the vaccinated and unvaccinated groups. Of the 429 registrants, 150 (35.0%) and 279 (65.0%) were vaccinated and unvaccinated, respectively. The average period from HPV vaccination to HPV testing was 102.7 months (8.6 years), with a median of 103 months (range 92–109 months). The HPV high‐risk infection rate was 21.3% (32/150) in the vaccinated group and 23.7% (66/279) in the unvaccinated group (P = 0.63). The HPV16/18 infection rate was 0% (0/150) in the vaccinated group and 5.4% (15/279) in the unvaccinated group, showing a significant difference (P = 0.0018), and the vaccine effectiveness was 100%. The cross‐protective type HPV31/45/52 infection rate in the vaccinated group was significantly lower than that in the unvaccinated group (3.3% vs. 10.0%, P = 0.013). There was no significant difference in the mean age at sexual debut and the number of previous sexual partners between the two groups. We have demonstrated the long‐term 9‐year effectiveness of the bivalent vaccine against HPV infection for the first time in Japan.     

儿童急性淋巴细胞白血病中RAS基因突变的检测及其临床意义

目的探讨RAS基因在儿童急性淋巴细胞白血病中的突变率及其临床意义。方法回顾性收集2015年1月至2020年1月于郑州大学第三附属医院收治的新确诊且完成二代测序的急性淋巴细胞白血病120例患儿病例资料，分析其临床及分子学特征，以及RAS基因突变对急性淋巴细胞白血病患儿总生存率的影响。结果120例患儿中，共有35例（29.2%）患儿伴有RAS基因突变，其中仅KRAS基因突变30例（25.0%），NRAS基因突变伴KRAS基因突变5例（4.2%）。全部NRAS基因突变及71%（25/35）KRAS基因突变位于第12、13号密码子。RAS基因突变在急性B淋巴细胞白血病中检出率为33.3%（35/105），未在急性T淋巴细胞白血病中检出。失访11例（9.2%），随访109例患儿中死亡16例（14.7%）。RAS基因突变患儿的2年总生存率低于RAS基因阴性患儿（P<0.05）。伴RAS基因突变急性淋巴细胞白血病患儿合并WT1基因过表达、初诊白细胞数计数>50×10⁹/L时，预后更差（P<0.05）。结论 RAS基因突变多发生于急性B淋巴细胞白血病，对预后有不良影响。     

血浆置换在儿童重症噬血细胞综合征中应用的疗效分析：前瞻性随机对照研究

目的探讨血浆置换在辅助救治儿童重症噬血细胞综合征（hemophagocytic syndrome，HPS）中的疗效和应用价值。方法采用前瞻性随机对照研究方法，选取2018年10月至2020年10月在湖南省儿童医院儿童重症监护室（pediatric intensive care unit，PICU）接受救治的重症HPS患儿40例为研究对象，将患儿随机分为置换组和常规组（n=20），常规组患儿进行病因和对症常规支持治疗，置换组在常规治疗的基础上加以血浆置换辅助治疗。对两组患儿的基本情况、治疗前后的临床症状体征、主要实验室检查指标、疗效和预后情况进行比较分析。结果两组患儿治疗前在性别、年龄、入院前病程、病因构成、小儿危重病例评分、器官或系统功能累及情况等指标上比较差异均无统计学意义（P>0.05），在实验室指标的比较上差异亦均无统计学意义（P>0.05）。两组患儿治疗7 d后，临床症状体征均有所缓解和改善。治疗后置换组C反应蛋白、降钙素原、血清铁蛋白、丙氨酸氨基转移酶、总胆红素水平均低于常规组（P<0.05）。置换组的PICU住院时间较常规组明显缩短，总有效率较常规组明显提高（P<0.05），但两组总的住院时间和3个月病死率比较差异无统计学意义（P>0.05）。结论血浆置换在辅助治疗儿童重症HPS上的疗效优于常规治疗，能改善患儿的临床症状体征和部分实验室指标，缩短PICU住院时间，可能成为治疗儿童重症HPS的一项有效辅助治疗方法。     

宫颈液基细胞学检查与高危型HPV检测用于宫颈癌前病变并发感染早期筛查的对比研究

目的:探讨宫颈液基细胞学检查与高危型HPV检测用于宫颈癌前病变并发感染的早期筛查的临床价值。方法:通过分析2014年4月至2015年4月在我院进行宫颈癌前病变筛查的480患者的临床资料,全部患者行宫颈液基细胞检查和高危型HPV检测,检查结果呈异常的患者行宫颈组织病理检查,观察两组临床疗效。结果:480例患者中TCT检查检出54例患者涂片异常,检出率为11.25%;高危型HPV检测结果显示76例患者高危型HPV感染呈阳性占15.83%;TCT检查和高危型HPV检查异常患者经宫颈组织病理学检查显示79例患者发生宫颈癌前病变,单纯TCT检查确诊率为77.14%,联合使用TCT检查和高危型HPV检测确诊率为96.20%,两种检测方法结果比较差异有统计学意义(P0.05)。结论:宫颈液基细胞联合高危型HPV检测可提高宫颈癌前病变并发感染的早期筛查率,对诊断宫颈癌变有一定的临床价值。     

唐氏综合征胎儿羊水外泌体miRNA差异表达谱分析

目的探讨胎儿羊水外泌体中miRNA在唐氏综合征（DS）胎儿生长发育中的作用。方法收集DS胎儿羊水和正常胎儿羊水，分别提取羊水外泌体miRNA。设置对照组：正常胎儿羊水外泌体miRNA；DS组：DS胎儿羊水外泌体miRNA。运用miRNA测序技术筛选出两组中差异表达的miRNA，并对差异表达的miRNA进行靶基因预测及功能分析（GO）和信号通路分析。从差异表达的miRNA中挑选3个与DS表型最相关的miRNA进行qPCR验证。通过双荧光素酶报告基因技术验证let-7d-5p对BACH1的靶向调控作用。结果和对照组相比，DS组中存在15个差异表达的miRNA，其中表达上调的miRNA有7个，表达下调的miRNA有8个。靶基因预测结果发现差异表达的miRNA可以靶向调控17种与DS相关的基因。GO分析发现靶基因主要功能与蛋白结合、蛋白转运、ATP结合、转移酶活性、突触等有关。Pathway通路分析发现富集显著的功能通路与神经系统发育有着密切的联系。qPCR验证结果发现与对照组相比，DS组中miR-140-3p、let-7d-5p水平显著降低（P < 0.05），与测序结果一致；而DS组中miR-4512水平较对照组显著增加（P < 0.05），与测序结果相反。双荧光素酶报告基因检测结果证实let-7d-5p可靶向调控BACH1的表达。结论羊水外泌体let-7d-5p可能通过调控BACH1的表达促进DS胎儿大脑氧化应激事件。     

γH2AX在HPV16阳性宫颈鳞癌组织中的表达及意义

目的 探讨γH2AX在HPV16阳性宫颈鳞癌组织中的表达及意义.方法 对74例宫颈鳞癌组织通过DNA提取,PCR检测,分析HPV的感染情况并筛选HPV16阳性宫颈癌组织;进而对HPV16阳性的宫颈癌石蜡组织连续切片HE染色明确组织型别,进行HPV16 DNA原位杂交检测HPV定位、免疫组化检测γH2 AX和p16蛋白的表达;最后选取30例典型的包括从正常宫颈组织、宫颈上皮内瘤变到宫颈原位癌渐变的组织切片,对比HPV DNA定位与γH2 AX和p16蛋白的表达,分析γH2 AX作为HPV感染导致宫颈癌发生过程中生物标记物的可行性.结果 经PCR扩增证明HPV感染率为98.65％,HPV16是最常见的型别占74.32％;原位杂交结果显示正常宫颈组织和CIN Ⅰ检测不到HPV16 DNA的存在,CINⅡ中HPV主要为游离型存在,随着宫颈病变的加重,HPV16 DNA逐渐出现整合形式;p16和γH2AX的表达均随着宫颈上皮病变级别的增加其阳性表达率增高,HPVDNA和γH2AX的表达均以颗粒细胞层和棘细胞层为主,定位具有一致性,HPV DNA和p16的表达定位不具有一致性.结论 γH2AX作为DNA损伤激活的重要蛋白,可作为HPV16感染致宫颈癌发生过程中的生物标记物.