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.

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。

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.

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.

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.

EXTRACAPSULAR SPREAD IN IPSILATERAL NECK METASTASIS:AN IMPORTANT PROGNOSTIC FACTOR IN LARYNGEAL CANCER

Objective To evaluate the impact of extracapsular spread （ECS） in ipsilateral neck metastasis on prognosis and its related factors in laryngeal cancer. Methods The study included 184 patients who underwent laryngectomy and simultaneous radical or modified radical neck dissection between January 1994 and December 1997 for laryngeal cancer. All of them had a complete 5-year follow-up. We used transparent lymph node detection and continuous slicing method on all neck dissection specimens. Kaplan-Meier model was used for survival analysis and the log-rank test was used to assess significance. Reults We found pathological neck metastases in 80 patients. Among them, 26 cases （32. 5% ） had ECS in ipsilateral neck. ECS incidence increased with advanced pathological N （pN） stages （pNl 3.7%, pN2a 25.0%, pN2b 50. 0%, and pN2c 55.6% ; P =0. 001）. ECS incidence also increased with number of positive nodes （1 positive node 8.6%, 2 positive nodes 33.3%, 3 and more positive nodes 66. 7% ; P 〈0. 001 ）. Incidences of contralateral neck metastases and ipsilateral neck recurrence in patients with ECS were higher than those in patients without ECS （46.2% vs. 24. 1%, P=0.046; 34.6% vs. 7.4%, P =0.002）. The 5-year survival rate of patients with ECS was significantly lower than that of patients without ECS （23.1% vs. 57.4%, P =0. 013）. Conclution ECS is an important prognostic factor in laryngeal cancer. Patients with ECS have a higher incidence of contralateral neck metastasis, so bilateral neck dissection should be selected.

Reconstruction of laryngeal function in subtotal laryngectomy with preservation of monoarytenoid cartilage

Objective: To evaluate subtotal laryngectomy with preservation of monoarytenoid cartilage to re-construct laryngeal function in the treatment of advanced laryngeal cancer. Methods: We retrospectively re-viewed 48 patients with advanced laryngeal cancer (T3 or T4 on the primary site) treated by subtotal laryngec-tomy with preservation of monoarytenoid cartilage to reconstruct laryngeal function. Eighteen of them under-went neck dissection. Ipsilateral recurrent laryngeal nerve was preserved during this surgery. The cricoid cartilage was anastomosed to hypopharynx accordingly. Results: The overall 3- and 5-year survival rates were 81.25% (39/45) and 66% 67% (32/48), respectively. All patients had good phonation and swallowing function after surgery. Decannulation rate was 93. 75%. Conclusion: Subtotal laryngectomy with preserva-tion of monoarytenoid cartilage is satisfactory for treatment of elected advanced laryngeal cancers. The effect of this surgery is more satisfactory than that of the near-total laryngectomy (Pearson''s technic).

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.

结合术前中性粒细胞与淋巴细胞比值、血小板与淋巴细胞比值构建喉恶性肿瘤进展风险的列线图模型

目的探究术前血小板与淋巴细胞比值(platelet-lymphocyte ratio,PLR)及中性粒细胞与淋巴细胞比值(neutrophil-lymphocyte ratio,NLR)在预测喉癌恶性肿瘤程度的价值并构建列线图模型进行验证。方法回顾性分析2016年1月~2022年6月在宁波市医疗中心李惠利医院接受喉部手术的339例患者临床、组织病理学和实验室资料,分为喉部良性病变组(n=113)、喉部癌前病变组(n=105)和喉恶性肿瘤(n=121)组,研究三组患者通过单因素分析和多因素Logistic回归分析研究喉部恶性病变的相关影响因素,采用R软件构建预测喉癌恶性肿瘤进展风险的列线图模型并进行内部验证。结果单因素分析显示年龄>60岁、吸烟、高PLR及高NLR与喉部病变程度有关(P<0.05)。多因素Logistic回归分析显示年龄>60岁、吸烟、高PLR及高NLR是喉部恶性病变的独立影响因素(P<0.05)。列线图模型C-index=0.809,95%CI=0.733、0.885,P<0.05,该列线图模型具有很强的预测能力。结论术前NLR、PLR、血小板分布宽度(PDW)是影响喉部恶性病变的因素,其中NLR高水平、PLR高水平在用于构建预测喉恶性肿瘤进展的列线图模型上准确性较强,有效的运用此模型可助于临床医师通过预测喉恶性肿瘤趋势为患者提供及时有效的用于早期筛查和疾病诊断的个性化方案。

喉神经内分泌癌1例

1临床资料患者,男,60岁,因“咽部不适1年”入院。1年前患者无明显诱因出现咽喉部不适和异物感,咽痛、咳嗽、咳粘痰,无呼吸困难,无吞咽阻挡感,药物治疗未见明显好转,为明确诊断就诊于济宁医学院附属医院耳鼻咽喉头颈外科门诊,电子喉镜检查可见会厌喉面右侧缘一不规则肿物,呈淡蓝色菜花样,黏膜表面不光滑(图1A)。喉部MRI示会厌右缘肿物,大小约9 mm×10 mm×14 mm(图1B、1C)。于2020-11-16全麻下行支撑喉镜下会厌肿物切除术,将会厌喉面肿物用电钩切除送快速病理,结果为(会厌)恶性肿瘤,但家属拒绝扩大手术切除范围。术后病理结果为非典型类癌(图1D)。

去整合素金属蛋白酶10和高迁移率族蛋白B1在声门型喉癌患者中的表达及预后分析

目的探讨去整合素金属蛋白酶10(a disintegrin and metalloprotease 10,ADAM10)和高迁移率族蛋白B1(high mobility group box-1 protein,HMGB1)与声门型喉癌患者病理特征及预后关系分析。方法回顾性收集2017年3月~2020年12月于南京医科大学附属南京医院确诊及治疗的声门型喉癌患者50例(观察组),另取相对喉癌组织切缘0.5cm以上部位标本作为对照组。观察并比较ADAM10和HMGB1在两组中的阳性表达率,分析其阳性表达与声门型喉癌患者的病理特征关系。单因素分析影响声门型喉癌预后的危险因素,Cox多因素回归分析声门型喉癌患者不良预后的独立危险因素。结果ADAM10和HMGB1在观察组的阳性表达率均高于对照组,差异均有统计学意义(P<0.05)。声门型喉癌组织中的ADAM10与淋巴结转移和T分级差异比较有统计学意义,而与年龄、性别、饮酒史、吸烟史、分化程度差异比较无统计学意义(P>0.05);HMGB1与分化程度差异比较有统计学意义(P<0.05),而与年龄、性别、饮酒史、吸烟史、淋巴结转移、T分级差异比较无统计学意义(P>0.05)。单因素分析结果表明,淋巴结转移、T分级、分化程度、ADAM10、HMGB1是患者预后的影响因素。Cox多因素回归分析结果表明,淋巴结转移、T3+T4分级、低分化程度、ADAM10阳性、HMGB1阳性为声门型喉癌患者预后不良的独立影响因素(P<0.05)。结论ADAM10和HMGB1可作为声门型喉癌不良预后的风险评估指标。

基于磷脂酰肌醇3激酶/蛋白激酶B/雷帕霉素靶蛋白信号通路小干扰RNA沉默微小RNA-373对喉癌细胞的影响

目的 探讨磷脂酰肌醇3激酶(Phosphatidylinositol 3 kinase,PI3K)/蛋白激酶B(Protein kinase B,Akt)/雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路小干扰RNA(Small interfering RNA,siRNA)沉默微小RNA-373(Microrna-373,miR-373)对喉癌细胞生物学行为的影响。方法 喉癌TU212细胞株经常规培养后分为空白组、空白转染组、过表达组和沉默组,四组细胞分别培养。检测各组细胞增殖、凋亡、迁移、侵袭能力及PI3K/AKT/mTOR通路蛋白表达。结果 与过表达组相比,沉默组miR-373、P13K、AKT、mTOR表达量较低(P<0.05);沉默组24、48、72 h细胞增殖率较低,72 h细胞凋亡率较高(P<0.05);沉默组细胞迁移率较少、侵袭数较少(P<0.05)。结论 沉默miR-373可能通过作用于PI3K/AKT/mTOR信号通路,下调P13K、AKT、mTOR表达,抑制喉癌细胞增殖、迁移、侵袭,促进凋亡。

喉肿瘤患者术前C反应蛋白检测的临床意义

C反应蛋白（C-reacitve protein,CRP）是第一个被确认的急性时相反应蛋白,在急性创伤、感染、炎性疾病及肿瘤发生时,在炎性因子尤其是IL-6的刺激下由肝细胞产生。近年来大量研究表明CRP在肺癌、结直肠癌、食管癌等肿瘤患者的发病及预后方面起着重要作用,但关于CRP与喉肿瘤的关系尚无讨论。

半坐下肢抬高位在喉梗阻强迫体位气管切开术中的应用

目的 探讨半坐下肢抬高位(V形体位)在喉癌引起的喉梗阻患者强迫体位气管切开术中的应用优势。方法收集2010年1月~2015年1月喉癌强迫体位喉梗阻在半坐位行气管切开术(半坐位组)患者20例,与2015年1月~2019年1月喉癌强迫体位喉梗阻在V形体位下行气管切开术(V形体位组)患者25例,进行临床对比分析。结果 两组患者手术均顺利完成,其中V形体位组4例出现术后并发症,半坐位组7例出现术后并发症,两组比较差异无统计学意义(χ^(2)=2.172,P>0.05)。V形体位组手术总时间及气管切开时间分别为(5.16±0.94)min、(3.44±0.77)min,半坐位组手术总时间及气管切开时间分别为(6.60±0.68)min、(5.00±0.79)min,两者相比差异有统计学意义(t分别为6.67、5.94,P<0.05)。结论 不能平卧的喉癌伴强迫体位喉梗阻患者可考虑采取V形体位行紧急气管切开手术。

喉神经内分泌癌临床病理学分析

神经分泌癌是一种由神经内分泌细胞分化而来的特殊的恶性肿瘤，尤其是喉神经内分泌癌（1aryngeal neuroendocrine carcinoma．LNEC）发生率极低。本病首先由Goldman（1969年）报道，目前国外共报道650例。近20年来随着诊疗技术的进步，本病的相关报道有所增多，但多属于个例报道．目前国内外对本病仍无系统的认识。

B细胞异位基因2在喉癌组织中的表达及临床意义

目的探讨喉癌组织中B细胞异位基因2(B cell translocation gene 2,BTG2)的表达与患者临床病理特征和预后的关系。方法收集川北医学院附属医院2015年1月~2018年8月手术切除的喉癌组织及癌旁组织各76例,采用免疫组化SP法检测喉癌组织及癌旁组织中BTG2的表达情况,并分析BTG2与喉癌临床病理因素及预后的关系。结果BTG2在喉癌组织及对应癌旁组织中均为弥漫性胞浆表达。喉癌组织中BTG2的阳性率为38.16%(29/76),低于癌旁组织中BTG2的阳性率90.79%(69/76),差异有统计学意义(χ2=45.957,P<0.001);BTG2表达与年龄(χ2=4.112,P=0.043)、分化程度(χ2=5.824,P=0.043)、颈部淋巴结转移(χ2=4.786,P=0.029)和复发(χ2=4.461,P=0.035)密切相关。Kaplan-Meier生存曲线显示BTG2阳性表达组生存率明显高于阴性表达组,单因素Log-rank检验分析显示临床分期、颈部淋巴结转移和BTG2表达与患者预后相关,进一步行多因素Cox风险比例回归模型分析发现颈部淋巴结转移和BTG2表达水平是影响喉癌预后的独立因素。结论BTG2在喉癌组织中的表达显著低于癌旁组织,且其表达与喉癌患者的年龄、分化程度、颈部淋巴结转移、复发及预后密切相关。BTG2在喉癌的发生发展中可能发挥重要作用,该蛋白有望成为评估喉癌预后的潜在指标。

喉鳞状细胞癌患者自噬相关微小RNA的鉴定及预后意义

目的鉴定喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)中自噬相关微小RNA(microRNA,miRNA),探讨其在肿瘤发展中的作用。方法数据库下载自噬相关基因及各个LSCC样本的miRNA表达谱,用limma包筛选差异表达的miRNA,并构建miRNA与自噬相关基因之间的共表达网络,从而筛选出自噬相关miRNA。Cox回归分析用于评估自噬相关miRNA在LSCC中的预后价值,建立基于自噬相关miRNA的风险指数模型以预测LSCC的预后。结果最终鉴定了6个关键自噬相关miRNA分别为hsa-miR-100-5p、hsa-miR-143-3p、hsa-miR-3607-5p、hsa-miR-454-3p、hsa-miR-455-5p、hsa-miR-99a-5p,均与LSCC的预后显着相关。富集分析表明所有的关键自噬相关miRNA在mTOR信号传导途径中均显著富集,利用多元Cox回归计算每个样品的风险指数,风险指数较低的患者具有更好的生存结果。结论关键自噬相关miRNA可以做为LSCC潜在的治疗靶标,基于自噬相关miRNA新构建的风险指数模型可以预测LSCC的预后。

Alveolar Rhabdomyosarcoma of the Laryngohypopharynx： A Case Report

Introduction Rhabdomyosarcoma is one of the most commonly seen soft-tissue sarcomas in children and teenagers, approximately accounting for half of all soft-tissue sarcomas. The tumor is usually found in head and neck, four limbs and urogenital system. However, the tumor orig- inating from the laryngohypopharynx is rare. In 2008, a patient with alveolar rhabdomyosarcoma of the laryngohypopharynx was admitted to our hospital. Clinical information of the disease was reported as follows, and clinicopathologic features and current therapeutic principle were discussed and summarized in combination with the review of literature, to facilitate the diagnosis and treatment of the patients with rhabdomyosarcoma of the laryngohypopharynx in future.

喉炎性肌纤维母细胞瘤诊断及治疗方法的探究

目的探讨喉炎性肌纤维母细胞瘤的诊断与治疗方法。方法回顾性分析4例喉炎性肌纤维母细胞瘤患者的临床病例资料,分析并总结其临床特征、治疗及预后情况。结果病变部位均为声带前端,声音嘶哑为主要症状,均于全麻下行支撑喉镜下喉显微CO2激光手术,病理示瘤组织细胞多呈梭形、短梭形、束状、旋涡状或编制装排列,异形明显。术后1个月嗓音功能恢复良好,3例(75%)声音嘶哑症状消失,1例(25%)明显改善。最长随访6年,均未复发及转移。结论喉炎性肌纤维母细胞瘤好发于声带,以病理及免疫组化结果为主要诊断依据,及早行根治性手术治疗是其治愈的关键。

LOSS OF HETEROZYGOSITY ON CHROMOSOME 13 IN SQUAMOUS CELL CARCINOMAS OF THE LARYNX

Objective: To locate lost region of tumor suppressor gene on chromosome 13q in squamous cell carcinoma of the larynx (LSCC) and to provide clues and evidence for discovering and locating new suppressor gene. Methods: Loss of heterozygosity (LOH) on chromosome 13q was analyzed in 58 LSCC patients by microsatellite polymorphic sequences in loci D13S765 (13q13), RB120 (13q142), D13S133 (13q143) and D13S318 (13q21) on chromosome 13 by PCR. Results: There weren't any LOH on chromosome 13q in 3 cases with preinvasive LSCC. Fortyfive percentage (24/53) of the 53 invasive LSCC cases showed LOH at one or more loci on chromosome 13q region. The highest percentage of LOH on chromosome 13q was 52% (22/53) at D13S765 locus. Conclusion: The deletion region on chromosome 13q was located near by D13S765 locus which is centromeric to RB1. In this region there is suppressor gene, which is related to the genesis and development of LSCC, possibly including RB1. The inactivation of these suppressor genes may be related to the genesis and development of invasive LSCC.