Circulating immune parameters-based nomogram for predicting malignancy in laryngeal neoplasm

BACKGROUND Malignancy prediction remains important to preoperative diagnosis and postoperative follow-up in laryngeal neoplasm.AIM To evaluate the circulating immune population and develop a nomogram for prediction of malignancy in patients with laryngeal neoplasm.METHODS A primary cohort of 156 patients was divided into laryngeal benign lesion,premalignant lesion and malignant lesion groups.Peripheral blood from patients was measured by blood routine test and flow cytometry.A nomogram was developed and applied to a validation cohort containing 55 consecutive patients.RESULTS Age,gender and seven circulating immune parameters exhibited significant differences between laryngeal benign lesion and premalignant lesion.The nomogram incorporated predictors,including gender,age,smoke index,proportions of monocytes,CD8+T cells,CD4+T cells,B cells and CD4/CD8+T cell ratio.It showed good discrimination between laryngeal premalignant lesion and malignant lesion,with a C-index of 0.844 for the primary cohort.Application of this nomogram in the validation cohort(C-index,0.804)still had good discrimination and good calibration.Decision curve analysis revealed that the nomogram was clinically useful.CONCLUSION This novel nomogram,incorporating both clinical risk factors and circulating immune parameters,could be appropriately applied in preoperative individualized prediction of malignancy in patients with laryngeal neoplasm.

Clinical significance of tumor-associated macrophage infiltration in supraglottic laryngeal carcinoma

Tumor-associated macrophages(TAMs) can elicit contrasting effects on tumor progression,depending on different tumor microenvironment.This study aimed to explore the correlation between TAM infiltration and clinicopathologic characteristics,metastasis,and prognosis of supraglottic laryngeal carcinoma.TAMs in intratumoral and peritumoral regions of 84 specimens of supraglottic laryngeal carcinoma tissues were detected by immunohistochemical staining with monoclonal CD68 antibody.The density of peritumoral CD68+ TAMs in recurrence cases(9/11) and in dead cases(17/23) were significantly higher than those in non-recurrence cases(33/73) and in survival cases(25/61),with significant differences(P = 0.024 and 0.007,respectively).The Kaplan-Meier survival analysis showed a significant relationship between the infiltration of both intratumoral and peritumoral CD68 + TAMs and the overall survival of patients.The 5year survival rate was significantly lower in the group with a high density of intratumoral CD68+ TAMs than in the group with a low density(39.6% vs.82.5%,P < 0.05).Similarly,the 5-year survival rate was significantly lower in the group with a high density of peritumoral CD68+ TAMs than in the group with a low density(50.6% vs.73.1%,P < 0.05).Cox regression analysis revealed that T classification,distant metastasis,and intratumoral or peritumoral CD68 + TAMs were independent factors for disease-free survival,whereas T classification and intratumoral CD68 + TAMs were independent factors for overall survival.The results indicate that TAM infiltration in supraglottic laryngeal carcinoma can be used to predict metastasis and prognosis and is an independent factor for prognosis.

Fiberoptic Intubation through Adapter Removable Supraglottic Airways;Comparison of the Air-Q ILA^TM, LMA Classic Excel^TM, and LMA Unique^TM

Study Objective: We studied the overall efficacy of fiberoptic aided intubation using three different supraglottic airways (SGA) as intubation conduits with a standard endotracheal tube (ETT) to determine which, if any, is superior as an intubation conduit. Design: After induction of general anesthesia, subjects were randomized to one of three groups: Air-Q ILATM, LMA Classic ExcelTM, and LMA UniqueTM. Subjects were intubated with a fiberoptic aided technique with continuous ventilation with FiO2 = 1.0 through one of these SGAs. The primary endpoint was the overall efficacy of the intubation procedure. In addition, the following data were collected: demographic data, intubation times, grade of view of the larynx, and a visual analog scale (VAS) score of difficulty as determined by the primary anesthesiologist performing the procedure. Data were analyzed using a Kruskal-Wallis one-way analysis of variance and Post hoc analysis was done using Dunn’s Multiple Comparison Test. Results: 126 total subjects were studied. Intubation success rates were 100%, 87.8%, and 95% with the Air-Q ILATM, LMA Classic ExcelTM, and LMA UniqueTM respectively. There was no significant difference among the three different SGAs when comparing the times to place the SGA (T1), the true intubating time (T2), the time to remove the SGA (T3), or the total time (T4). Data were also stratified by the grade of view of the larynx;all grade I views, grade II views, and grade III views were grouped together regardless of the type of the SGA used. The grade I view of the larynx group had significantly faster true intubation times (T2 = 75.1 sec, p = 0.01) and significantly lower VAS scores (VAS = 1.9, P = TM provides the best view of the larynx and is the easiest one to use as an intubation conduit.

Investigating the Fluid Seal of Supraglottic Airway Devices in Humans Using Indicator Dye via the Drainage Tube:A Potential Roadmap for Future Studies

Background: The fluid seal of supraglottic airway devices (SGA) protects the airway from fluid contamination. We evaluated the suitability of indicator dye placement in the upper digestive tract of anesthetized patients combined with fiberoptical tracing to investigate the fluid seal of SGA. Methods: Patients swallowed a capsule of indigo carmine green (ICG) prior to induction of anaesthesia. After induction of anesthesia, one of two different SGA (either an i-GelTM or an LMA-SupremeTM (LMA-S)) was inserted after randomization. Methylene blue stained normal saline was injected through the proximal opening of drainage tube during mechanical ventilation as well as spontaneous breathing. We monitored regurgitation of ICG with a flexible fiberscope (FO) inserted through the drainage tube and checked for the appearance of methylene blue in the mask bowl with the FO inserted through the airway tube. Results: In thirty-six patients with an i-GelTM and 37 with a LMA-S no regurgitation of ICG was observed at the level of the upper oesophageal sphincter (UES). Methylene blue stained saline was not visible in any patient during pressure-controlled ventilation, but was detected in two of the 36 patients with the i-GelTM during spontaneous breathing. Conclusion: Instilling dye through the drainage tube of SGA models with a built-in drainage tube represents a useful method to examine and to compare the fluid seal of different SGA. Our protocol presented in this study proved to be an easy and reproducible approach for future studies. Furthermore, the clinical results gained during this evaluation highlight the necessity for further investigations regarding the fluid seal competencies of SGAs in humans under clinical conditions.

喉NUT癌1例

1临床资料患者,女,19岁,因“咳血2个月”就诊于外院,支气管镜活检病理:(声门下)鳞状上皮乳头状增生,伴重度非典型性。病理会诊:鳞状上皮乳头状增生,伴中重度非典型性,未治疗。2022-09-09患者因Ⅱ度呼吸困难收入郑州大学第一附属医院。喉部增强CT示:喉咽部壁明显增厚,右侧壁软组织增强扫描边缘可见强化,相应喉腔狭窄,双侧颈部肿大淋巴结(图1A)。入院后患者再次无诱因出现呼吸困难,急诊行气管切开术,气管前淋巴结送病理,术后冰冻病理结果:高级别转移性癌,结合免疫组化,符合低分化鳞状细胞癌。

沉默miR-373对喉癌细胞增殖、凋亡能力的影响及作用机制研究

目的探究沉默RNA-373(miR-373)对喉癌细胞增殖、凋亡能力的影响及作用机制研究。方法将喉癌细胞分为对照组、过表达组、沉默组3组,并通过细胞转染建立稳定转染的过表达组和沉默组。采用MTT法检测细胞增殖能力,TUNEL法检测细胞凋亡能力,Transwell小室检测细胞侵袭,细胞划痕实验检测细胞迁移,蛋白质印迹法检测各组细胞Wnt/β-catenin信号通路β-连锁蛋白(β-catenin)、c-myc、细胞周期素D1(CyclinD1)、基质金属蛋白酶9(MMP-9)、Bc1-2、Bax蛋白表达量。结果与过表达组相比,沉默组miR-373表达量明显下降(t=15.062,P<0.05)。与过表达组相比,沉默组细胞凋亡率较高,细胞不同时间点增殖率较低(t=31.025、16.453、22.475、29.672,P<0.05);与过表达组相比,沉默组细胞侵袭能力、迁移数均较低(t=35.254、37.205,P<0.05)。与过表达组相比,沉默组β-catenin、c-myc、CyclinD1、MMP-9、Bc1-2蛋白表达量较低,Bax蛋白较高(t=4.218、5.307、4.609、5.005、4.328、3.984,P<0.05)。结论沉默miR-373可能通过促进Bax表达,抑制β-catenin、c-myc、CyclinD1、MMP-9、Bc1-2表达,阻滞Wnt/β-catenin信号通路,从而促进喉癌细胞凋亡,抑制喉癌细胞侵袭、增殖、迁移。

支撑喉镜CO_(2)激光术后复发的喉癌患者窄带成像特征分析

目的分析行支撑喉镜CO_(2)激光术后复发的早期声门型喉癌患者的窄带成像(NBI)内镜特征。方法回顾性分析2017年1月~2023年12月就诊于西安交通大学第二附属医院耳鼻咽喉头颈外科的Ⅰ~Ⅱ期声门型喉癌患者中,行支撑喉镜CO_(2)激光术后复发者。系统收集复发者术前、术后1个月、3个月、6个月等不同时间点的病史、治疗方式、喉镜检查、影像学检查和病理结果,总结复发者NBI内镜特征。结果18例行支撑喉镜CO_(2)激光术后复发的早期声门型喉癌患者,NBI内镜诊断喉癌复发的诊断率显著高于普通白光内镜(88.89%vs.55.56%,χ^(2)=4.985,P=0.026)。所有复发患者按ELS分型,上皮内乳头样毛细血管袢(intraepithelial papillary capillary loop,IPCL)均为垂直血管;按Ni分型,72.22%(13/18)IPCL为V型,27.78%(5/18)IPCL仅表现为可疑异常新生血管(点状或扩张弯曲)。术后随访时的NBI内镜特征显示:术后1个月、3个月、6个月,术区表面有异常IPCL的患者比例分别为0.00%、27.78%(5/18)、61.11%(11/18),差异有统计学意义(χ^(2)=16.164,P<0.001),即喉癌复发患者随访时间越长,NBI内镜下术区表面有异常IPCL的患者比例越高。结论对行支撑喉镜CO_(2)激光术后的早期声门型喉癌患者,当NBI内镜下出现V型IPCL或点状、扩张弯曲的可疑新生血管时,可能早期提示喉癌复发。

缺氧环境下GAPDH通过抑制铁死亡增强喉鳞状细胞癌恶性生物学行为和顺铂耐药性研究

目的探讨缺氧环境下磷酸甘油醛脱氢酶(GAPDH)调控喉鳞状细胞癌(简称喉癌)铁死亡、恶性生物学行为和顺铂耐药性中的作用。方法人喉癌细胞系Hep-2和TU212在体外进行培养,分别在缺氧(1%O_(2))和常氧(21%O2)条件下进行无血清培养。Hep-2细胞分为5个实验组:DMSO对照组、Erastin组、缺氧+Erastin组、缺氧+Erastin+NC-GAPDH组和缺氧+Erastin+si-GAPDH组,以研究细胞在不同实验条件下的反应。使用定量聚合酶链反应(qPCR)和蛋白免疫印迹法(WB)分析各组细胞系GAPDH的mRNA和蛋白表达水平。细胞计数试验(CCK-8)、集落形成实验和Transwell侵袭实验用于评估各组喉癌细胞活力、增殖能力和侵袭能力。CCK-8实验用来测定细胞对不同浓度顺铂(5、15、20μg/ml)的耐药性。试剂盒法评估细胞内丙二醛(MDA)和谷胱甘肽(GSH)的水平。WB检测谷胱甘肽过氧化物酶4(glutathioneperoxidase4,GPX4)和4-羟基壬烯醛(4-hydroxynonenal,4-HNE)蛋白表达水平。同时,使用Fe2+探针和活性氧簇(ROS)荧光探针检测细胞内Fe2+含量及ROS水平,透射电子显微镜用于观察线粒体形态变化,以评估铁死亡发生情况。结果与常氧条件相比,缺氧显著增强Hep-2和Tu212细胞中GAPDH mRNA和蛋白的表达(P均<0.05),在缺氧条件下,细胞的集落形成数量、迁移数量以及在15μg/ml和20μg/ml顺铂处理下的细胞活力显著上升,均高于常氧组(P均<0.05)。GAPDH被沉默后,缺氧引起的这些增强效果得到逆转。在常氧条件下,与DMSO对照组相比,Erastin处理显著降低Hep-2细胞的集落形成数量、迁移数量以及在15μg/ml和20μg/ml顺铂处理下的细胞活力(P均<0.05),但在缺氧条件下,Erastin的抑制作用有所减弱。此外,缺氧+Erastin+si-GAPDH组的集落形成数、迁移数以及在15μg/ml和20μg/ml顺铂处理下的细胞活力显著低于缺氧+Erastin+NC-GAPDH组(P均<0.05)。结论在缺氧环境下,沉默GAPDH能够逆转喉癌细胞铁死亡抑制过程,进而抑制其恶性生物学行为和顺铂耐药性。

The Prognostic Value of Pathological and Molecular Margins Marked by p53 and eIF4E in Laryngeal Carcinoma

Objective: To study the prognostic value of the pathological margin and molecular margin marked by eIF4E and P53 protein in laryngeal carcinoma. Methods: The prognostic value of pathological and molecular margin was studied in 253 cases and 67 cases respectively, the latter were pathological negative margin chosen from the former. Immunohistochemisty was used to detect the expression of eIF4E and p53 proteins. Results: The rate of pathological, p53 and eIF4E positive margins was 20.2%, 19.4% and 32.8% respectively. The recurrent rate of those with positive margins was higher than that of negative margins, which including pathological margin (70.6% vs 35.1%, P =0.0000), p53 margin (69.2% vs 33.3%, P =0.018) and eIF4E margin (63.6% vs 28.9%, P =0.018); The survival rate of those with negative margins was higher than those with positive margins, including pathological margin (the 5-year cumulative survival rate was 37.52% and 64.37% respectively, P =0.0023), p53 margin (the 5-year cumulative survival rate was 24.62% and 75.69% respectively, P =0.0012) and eIF4E margin (the 5-year cumulative survival rate was 43.31% and 77.52% respectively, P =0.0006). Conclusion: The prognosis of those with both pathological and molecular positive margins was worse than that of the negative margins; Both the eIF4E and p53 were useful markers to pick out the poor prognostic patients from those with pathological negative margin, and the former seemed to be more potential.

血清CC型趋化因子2、CC型趋化因子18与声门型喉癌患者临床病理参数和预后的关系

目的研究血清CC型趋化因子2(CC type chemotactic factor 2,CCL2)、CCL18与声门型喉癌患者临床病理参数和预后的关系。方法选择邯郸市中心医院和河北工程大学附属医院2015年8月~2018年12月收治的168例声门型喉癌患者为研究对象,受试者工作特征(ROC)曲线确定血清CCL2、CCL18最佳截点,将患者分为CCL2高表达组和低表达组、CCL18高表达组和低表达组,分析血清CCL2、CCL18水平与声门型喉癌患者临床病理参数的关系,采用Kaplan-Meier曲线和Log-Rankχ^(2)检验分析血清CCL2高表达组和低表达组、CCL18高表达组和低表达组的5年无病生存率,采用Cox回归模型分析声门型喉癌预后的影响因素,并分析血清CCL2和CCL18表达与肿瘤复发/转移的关系。结果根据ROC曲线计算得出CCL2、CCL18的最佳截点分别为100.81、218.99 pg/ml。相较于CCL2、CCL18低表达组,CCL2、CCL18高表达组T3~T4a级、N1~N3级、肿瘤低分化占比明显升高(P<0.05)。168例声门型喉癌患者随访5年失访8例,完成随访160例。67例发生复发/转移,39例因复发/转移而死亡,肿瘤复发/转移率为41.88%(67/160),无病生存率为58.13%(93/160)。Kaplan-Meier生存曲分析显示,血清CCL2、CCL18高表达组5年无病生存率均明显低于血清CCL2、CCL18低表达组(P<0.05)。Cox回归分析显示,T分级升高、颈淋巴结复发、N分级升高、咽喉部复发、CCL2高表达、CCL18高表达是声门型喉癌预后不良的危险因素(P<0.05)。分析血清CCL2和CCL18表达与肿瘤复发/转移的关系发现,CCL2和CCL18均高表达时其复发/转移率明显高于CCL2和CCL18均低表达、CCL2低表达且CCL18高表达、CCL2高表达且CCL18低表达,差异比较有统计学意义(χ^(2)=10.450,P=0.015)。结论声门型喉癌患者血清CCL2、CCL18高表达与T分级、N分级、肿瘤低分化和预后不良显著相关。

术前静脉期CT影像组学列线图预测喉鳞癌患者预后

目的:研究增强CT影像组学术前预测喉鳞癌术后复发的价值。方法:收集本院64例经术后病理证实为喉鳞癌的患者作为训练组;将TCGA数据库中下载的39例患者作为验证组,基于患者术前CT增强静脉期图像,利用3D-slicer软件提取喉鳞癌全域影像组学特征。在训练组中,应用单因素COX及LASSO回归确定复发相关的最佳组学特征,构建影像组学风险评分对喉鳞癌患者预后分层;基于多因素COX回归构建预测1年、3年无进展生存的列线图。采用受试者工作特征曲线(ROC)评估影像组学风险评分的诊断效能。最后采用相同阈值对验证组患者进行模型验证。结果:训练组确定4个影像组学特征与复发显著相关,构建影像组学评分模型,Kaplan-Meier曲线显示训练组和验证组低风险组(组学评分<0.7)无进展生存时间均显著长于高风险组(P<0.001,P=0.025)。多因素COX回归研究表明影像组学评分是喉鳞癌术后复发的独立危险因素。在训练组中列线图预测喉鳞癌患者三年术后无进展生存C指数为0.816,影像组学模型C指数为0.795。在验证组中列线图C指数为0.858,影像组学模型C指数为0.795。结论:基于术前CT增强图像提取的喉鳞癌组学特征并建立列线图模型,可用于术前预测患者术后复发,且诊断效能高于临床常用TNM分期,可作为临床诊疗决策依据。

Pathologic features of occult lymphatic metastasis in supraglottic carcinoma

OBJECTIVE: To study the pathologic features of occult lymphatic metastasis in supraglottic carcinoma. METHODS: Serial sections of 153 neck dissection specimens in 100 patients with supraglottic carcinoma were evaluated under the microscope. RESULTS: In 100 patients, 38 had occult metastatic lymph nodes. 51 metastatic lymph nodes were found in pathology, and their sizes ranged from 0.5 cm to 2.6 cm (average 1.1 cm). The distribution of 51 lymph nodes was 1 in level I (2%), 37 in level II (73%), 12 in level III (24%), and 1 in level IV (2%). Among the 51 nodes, 21 (41%) were early stage, 18 (35%) were growth stage, 7 (14%) were tull stage, and 5 (10%) were extracapsular stage. The differentiation degree and appearance of supraglottic carcinoma was not directly related with occult metastasis. CONCLUSION: The occult metastatic rate of supraglottic carcinoma is high, and selective neck dissection may be necessary.

经口低温等离子射频消融在早期声门上型喉癌治疗中的有效性及安全性分析

目的:探究经口低温等离子射频消融(RFA)在早期声门上型喉癌治疗中的有效性及安全性。方法:选取2018年3月—2022年3月焦作市人民医院收治的102例行早期声门上型喉癌患者作为研究对象,根据治疗方式分组,分为对照组(57例)和观察组(45例)。其中对照组采用传统手术治疗,观察组采用经口低温等离子RFA治疗。比较两组患者围术期指标、黏膜恢复时间;比较术前、术后嗓音声功能指标、嗓音障碍评分(VHI)情况;比较术前及术后3 d、6 d、10 d的视觉模拟评分(VAS),并记录术后并发症和1年内复发率情况。结果:观察组手术时间、住院时间、黏膜恢复时间均短于对照组,且术中出血量低于对照组,差异有统计学意义(t=40.122、43.548、20.302、11.064,P<0.05);术后,观察组谐噪比高于对照组,且基频微扰、振幅微扰低于对照组,差异有统计学意义(t=8.129、6.539、12.726,P<0.05);观察组VHI评分均低于对照组,差异有统计学意义(t=19.086、27.101、23.151,P<0.05);观察组术后3 d、6 d、10 d VAS评分低于对照组,差异有统计学意义(t=3.969、6.732、16.645,P<0.05);观察组术后并发症低于对照组,差异有统计学意义(χ^(2)=4.157,P<0.05);两组患者1年内术后复发率比较,差异无统计学意义(χ^(2)=0.638、1.611,P>0.05)。结论:经口低温等离子RFA在早期声门上型喉癌治疗中具有手术时间短、出血量少、创伤性小的特点,能减轻患者疼痛情况,并有利于患者黏膜及嗓音恢复,降低嗓音功能障碍,减少并发症的发生。

ANALYSIS OF C-HA-RAS GENE AMPLIFICATION AND MUTATION IN LARYNGEAL CARCINOMA

In order to study the altered molecular events during laryngeal carcinogenesis and elucidate the role of Haras oncogene amplification and mutation. we have examined their profile by polymerase chain reaction (PCR) and selective oligonucleotide hybridization. We analyzed the mutational status of codon 12 of Haras in 22 laryngeal carcinomas and 10 normal tissues. and found that 7 of 22 laryngeal carcinomas contained a Ha-ras mutation at codon 12. The frequency of mutation was 32%. None of the normal tissues revealed mutation. Moreover. no amplification was found in cancers when compared to the normal. Ourfindings indicated that the activated Ha-ras gene existed in laryngeal carcinoma. and activation of the Haras gene by mutation at codon 12 might play a key role in laryngeal carcinogenesis.

幼年型复发性呼吸道乳头状瘤病中抑制性细胞因子与疾病严重程度的相关性研究

目的探讨免疫调节性细胞因子在幼年型复发性呼吸道乳头状瘤病(juvenile onset recurrent respiratory papillomatosis,JORRP)患者外周及病变局部的表达变化,分析其与JORRP疾病严重程度的相关性。方法采集JORRP患者外周血,同时采集性别、年龄相匹配的健康儿童的外周血作为对照组;收取JORRP患者病变组织,同时采集扁桃体切除术患者的扁桃体前腭舌弓黏膜上皮组织作为对照组。ELISA检测血浆样本中细胞因子浓度,并对组织样本进行mRNAseq检测及RT-PCR验证分析。进一步将细胞因子表达水平和疾病严重程度进行相关性分析。结果JORRP患者血浆中白细胞介素10(IL-10)浓度显著高于对照组(P<0.05),且患者血浆中IL-10表达与初次发病年龄呈负相关(r=-0.3307,P<0.05)。IL-10 mRNA表达水平在侵袭性组较非侵袭性组显著升高(P>0.05)。JORRP患者血浆TGF-β浓度与采样时手术次数呈正相关(r=0.5144,P<0.05),且侵袭性组TGF-β浓度较非侵袭性组显著增高(P<0.05)。TGF-βmRNA表达水平在JORRP患者较对照组显著增高(P<0.001)。结论JORRP患者中抑制性免疫调节因子IL-10和TGF-β高表达且与疾病严重程度正相关。

GPRC5A在喉癌发生中的生物学作用及机制研究

目的探讨G蛋白偶联受体C家族5A(Gprotein-coupled receptor family C,member 5,group A,GPRC5A)在喉癌组织中的表达特征,分析其在喉癌发展进程中的作用及机制。方法选取河北医科大学第二医院收治的32例经病理检查确定为喉鳞状细胞癌患者的喉癌组织及癌旁组织,采用免疫组织化学方法检测GPRC5A在喉癌组织及癌旁组织中的表达,并分析其表达特征与患者临床资料的相关性。采用定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)方法对GPRC5A在喉癌组织与癌旁组织的表达进行组织水平的验证,在人支气管上皮细胞系BEAS-2B、喉癌细胞系TU686及下咽癌细胞系Fadu中进行细胞水平的验证。构建GPRC5A过表达质粒p-GPRC5A,转染至喉癌TU686细胞中采用cell counting kit 8、Transwell方法检测GPRC5A基因过表达对喉癌TU686细胞增殖、侵袭及迁移的影响。Western blot检测GPRC5A基因过表达对细胞凋亡相关蛋白表达的影响。Western blot验证GPRC5A基因过表达对表皮生长因子受体(epidermal growth factor receptor,EGFR)/STAT3(epidermal growth factor receptor/signal transducer and activator of transcription 3)通路的影响。结果免疫组织化学结果表明,GPRC5A在喉癌组织中的阳性表达率显著低于癌旁组织(χ^(2)=14.190,P<0.001)。GPRC5A在不同肿瘤分期、淋巴结转移、分化程度喉癌组织中表达差异无统计学意义(P>0.05)。GPRC5A在喉癌组中的表达显著低于癌旁组(t=3.175,P=0.003)。GPRC5A在喉癌TU686和Fadu细胞中表达为BEAS-2B的(0.73±0.08)和(0.78±0.04)倍,差异有统计学意义(F=9.060,P=0.015);CCK8结果表明,48 h后p-GPRC5A组的OD值为0.76±0.03,显著低于NC组1.20±0.01和对照组1.30±0.08(F=69.970,P<0.001);Transwell实验表明,p-GPRC5A组细胞迁移的细胞数为138.70±10.97,显著低于对照组251.3±16.9和NC组247.7±17.5(F=5731.100,P<0.001)。p-GPRC5A组细胞侵袭的细胞数为113.00±10.21,显著低于对照组193.3±010.02和NC组190.00±7.90(F=8894.100,P<0.001)。Western blot结果表明,与对照组相比,Caspase3蛋白在p-GPRC5A组表达显著升高(F=78.880,P<0.001),Bcl-2蛋白在p-GPRC5A组的表达显著降低(F=125.820,P<0.001)。EGFR和p-STAT3蛋白在p-GPRC5A组的表达显著降低(F=27.573,P=0.001;F=60.614,P<0.001)。结论GPRC5A在喉癌组织及细胞系中低表达。GPRC5A基因过表达可抑制喉癌细胞增殖,迁移和侵袭,促进细胞凋亡发生,并抑制EGFR/STAT3通路的激活。

微RNA-509-3p/干扰素刺激基因15轴对喉癌细胞血管生成及血管内皮生长因子蛋白表达的影响

目的探究微RNA(miR)-509-3p/干扰素刺激基因15(ISG15)轴对喉癌细胞血管生成及血管内皮生长因子(VEGF)蛋白表达的影响。方法2022年1―9月,将喉癌M4E细胞分为Con组、miR-NC组、miR-509-3p组、pcDNA-ISG15组。逆转录聚合酶链式反应(RT-PCR)检测细胞中miR-509-3p表达;细胞计数试剂盒(CCK8)检测细胞增殖能力;Transwell检测细胞侵袭能力;划痕实验检测细胞迁移能力;流式细胞仪检测细胞凋亡率;Matrigel体外成管实验检测血管生成能力;蛋白质印迹法检测细胞中ISG15、VEGF蛋白表达;萤光素酶活性报告检测miR-509-3p和ISG15的靶向关系。结果与喉上皮细胞NP69中miR-509-3p表达(1.01±0.13)μg/L相比,喉癌细胞SCC-40、SCC-2、M4E中miR-509-3p表达[(0.61±0.08)、(0.45±0.07)、(0.18±0.07)μg/L]均降低,且M4E细胞中miR-509-3p表达低于SCC-40、SCC-2细胞(P<0.05)。与Con组相比,miR-509-3p组细胞中miR-509-3p表达增加[(1.17±0.05)μg/L比(118±0.03)μg/L];Con组细胞中miR-509-3p表达与miR-NC组相比差异无统计学意义(P>0.05)。与Con组相比,miR-509-3p组细胞增殖(1.71±0.02比3.09±0.07)、侵袭(55.33±2.52比150.67±2.52)、迁移(30.58±2.08比98.33±1.53)、形成小管数量(61.28±2.15比135.62±2.54)及细胞中ISG15(0.24±0.03比1.11±0.13)、VEGF(0.35±0.02比0.99±0.04)蛋白表达均降低,细胞凋亡率[(18.76±0.18)%比(5.61±0.08)%]增加(P<0.05);Con组细胞增殖、侵袭、迁移、形成小管数量、凋亡率及细胞中ISG15、VEGF蛋白表达和miR-NC组相比差异无统计学意义(P>0.05);与miR-509-3p组相比,pcDNA-ISG15组细胞增殖(3.25±0.06)、侵袭(144.00±2.65)、迁移(89.34±5.13)、形成小管数量(131.56±2.08)及细胞中ISG15(0.83±0.05)、VEGF蛋白(0.89±0.03)表达增加,细胞凋亡率降低(6.85±0.06)%(P<0.05)。结论过表达miR-509-3p可抑制喉癌细胞血管生成及VEGF蛋白表达。

幼年型复发性呼吸道乳头状瘤病患者生长发育、智力水平及心理状态的临床观察

目的探讨幼年型复发性呼吸道乳头状瘤病(juvenile-onset recurrent respiratory papillomatosis,JORRP)对生长发育、智力水平及心理状态的影响。方法回顾性分析2023年1~12月就诊于首都医科大学附属北京同仁医院的40例JORRP患者(观察组)与同期体检的40名健康青少年(对照组)的临床资料。应用体重指数(BMI)、瑞文标准推理测验(Raven’s standard progressive matrices,SPM)、自我缺陷感量表(the feelings of inadequacy scale,FIS)评价入组人群的生长发育、智力水平及心理状态。结果观察组BMI为(21.320±3.609)kg/m²,对照组为(22.250±2.685)kg/m²,两组间差异无统计学意义(U=668.000,P>0.05)。SPM分数经过标准表换算后得出,观察组智力水平良好者占5%(2/40)、中等者占55%(22/40)、中下者占35%(14/40)、低下者占5%(2/40),对照组智力水平良好者占5%(2/40)、中等者占90%(36/40)、中下者占5%(2/40),两组间SPM总分、A组知觉辨别能力、D组系列关系能力及E组抽象推理能力得分差异有统计学意义(P<0.05),且两组智力水平均与生活地区相关(|r|>0.3,P<0.05)。观察组与对照组FIS分数差异有统计学意义(86.650±21.680 vs.62.250±9.446,U=270.000,P<0.001),且观察组的初次发病年龄与其自卑感呈负相关(|r|>0.3,P<0.05)。观察组手术次数与智力水平、FIS得分无相关性(|r|<0.3,P>0.05)。结论JORRP不影响患者的生长发育,但能在一定程度上增加患者的自卑感,对患者心理健康造成负面影响。

Prognostic Value of Ki67 and VE6F Expression in Laryngeal Squamous Cell Carcinoma

OBJECTIVE To investigate the prognostic value of measuring Ki67 and VEGF expression in laryngeal squamous cell carcinoma (LSCC). METHODS The expression of Ki67 and VEGF in 32 LSCC tissues was studied by immunohistochemical staining. Of these cases, 5 recurred (recurrent group), 3 cases metastasized (metastatic group), 8 cases died (deceased group) and 24 cased survived (survival group) over a 3 year period of follow-up after their operation. RESULTS The expression of Ki67 and VEGF in the deceased group was higher compared to that in the survival group (P<0.01). Overexpression of Ki67 was found in the recurrent group and in the metastatic group (P< 0.05). VEGF expression was higher in the recurrent group than in the non recurrent group (P<0.05). With Cox-regression of multivariate analysis, Ki67 (RR:3.236; P=0.001), the clinical T stage (RR: 1.382; P=0.029) and metastasis in lymph nodes (RR:0.316; P=0.033) were shown to be independent prognostic factors for survival of LSCC patients. CONCLUSION Ki67 and VEGF expression is related to the prognosis of LSCC. Overexpression of the 2 markers indicated poor prognosis of the disease, a finding which might be helpful for the treatment of laryngocar-cinoma.

Effect of DRB on the Biological Characteristics of Human Laryngeal Carcinoma Hep-2 Cell Line

In order to study the effect of 5, 6-Dichloro-l-13-D-ribofuranosyl-benzimidazole （DRB） on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry （FCM） and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was （0.68±0.19）%, （1.95±0.12）%, （8.51±0.26）%, （11.26±0.17）% and （14.99±0.32）%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.