遗传性出血性毛细血管扩张症相关ACVRL1基因新突变——c.1246+5G>C

目的　遗传性出血性毛细血管扩张症（hereditary hemorrhagic telangiectasia，HHT）是一种遗传性系统性血管性疾病。本研究拟对1例HHT家系先证者临床特征及致病基因进行分析。方法　收集1例HHT家系先证者，对其行表型和基因突变进行分析。应用PCR扩增测序的方法对该病相关基因全部编码区进行序列分析，并应用NetGene2和minigene方法对发现的变异功能进行分析。结果　先证者 具有鼻出血、皮肤毛细血管扩张及家族史，符合HHT诊断；基因检测发现ACVRL1 基因c.1246+5G>C变异，为未报道的新突变。NetGene2软件分析发现其可影响外显子8的剪接。Minigene分析发现突变型在转录时外显子8被略过，导致ACVRL1 基因功能异常。结论　本研究在1例HHT家系中发现ACVRL1 基因外显子8附近一个新突变—c.1246+5G>C，支持ACVRL1基因外显子8区域为中国HHT的突变热点。     

遗传性出血性毛细血管扩张症临床及基因检测综合诊断的应用

目的　对3例遗传性出血性毛细血管扩张症（hereditary hemorrhagic telangiectasia，HHT）病例的临床和基因分析，探索临床及基因检测结合方法对该病的意义。方法　收集2例HHT家系，1例疑似病例，并进行临床和基因特征的分析；应用PCR-测序的方法对内皮素（endoglin，ENG）、激酶1样激活素受体（activin A receptor type II-like 1，ACVRL1）、SMAD4和骨形成相关蛋白9（bone morphogenetic protein 9，BMP9）基因全部编码区进行序列分析；结合临床特征及基因检测结果进行综合诊断。结果　基因检测发现两HHT家系先证者分别携带ENG基因的c.772delT（p.Tyr258Thrfs*101）突变和ACVRL1基因的c.1232G>A（p.Arg411Pro）突变，明确病因，并对疾病进行分类诊断。对1例疑似病例基因检测未发现基因突变，综合临床和基因检测分析，定义为不能诊断HHT。结论 HHT的临床和基因相结合可确定疾病的病因并分类诊断，可发现早期的仅具有鼻出血等单一症状的患者，可发现无症状基因突变携带者，便于疾病的早期预防，避免严重并发症发生。     

遗传性出血性毛细血管扩张症的基因检测和遗传咨询

遗传性出血性毛细血管扩张症（hereditary hemorrhagic telangiectasia，HHT）是一种罕见的系统性血管性疾病，主要症状来源于动静脉吻合畸形，表现为反复的鼻出血、皮肤黏膜毛细血管扩张、动静脉畸形，可合并严重的并发症。然而，由于HHT患者发病年龄和表型变异大，临床对于该病的早期诊断存在困难。HHT是一种常染色体显性遗传性疾病，常见的致病基因包括ENG、ACVRL1和SMAD4，这几种基因的突变和90%以上的患者发病有关。随着对HHT分子致病机制认识的不断深入，使得基因检测成为HHT辅助诊断的工具。基因检测和遗传咨询不仅可以确认临床HHT的诊断，并可对其进行分型，通过家系成员基因筛查和遗传咨询，明确无症状 感染者、高危个体以及非高危个体。本文将对基因检测和遗传咨询在HHT高危人群的健康管理及明确诊断中的应用进行总结，为其广泛应用提供相应准则。     

遗传性出血性毛细血管扩张症致严重鼻出血的早期基因诊断

目的:探讨遗传性出血性毛细血管扩张症(HHT)致严重鼻出血的早期基因诊断。方法:对2个HHT家系共23例成员进行详细的临床检查及评估,提取外周静脉血DNA,聚合酶链反应扩增目的基因ENG和ACVRL-1,测序并进行序列分析,判断突变的致病性。结果:NMG-1家系采集到静脉血标本的11例中有6例携带ACVRL-1基因的错义突变c.263A>G。GD-2家系采集到静脉血标本的12例中有5例携带ACVRL-1基因的错义突变c.199C>G。有鼻出血病史者突变基因检出率为100%,无鼻出血者史突变基因检出率为25%。结论:基因诊断极高的灵敏度和特异性在HHT早期诊断中具有重要的应用价值,可成为临床常规检测项目。     

遗传性出血性毛细血管扩张症一家系ALK1基因突变研究和临床微创治疗

目的研究遗传性出血性毛细血管扩张症（hereditary hemorrhagic telangiectasis,HHT）一家系活化索受体激酶1（activin receptor-like kinase-1,ACVRL1或ALK1）基因突变情况,评估双极电凝治疗鼻出血的疗效。方法收集1例HHT家系,采用聚合酶链反应（polymerase chain reaction,PCR）扩增其成员ALK1基因全部编码外显子及侧翼序列,对PCR产物进行序列分析,以其健康者和100例无血缘关系的正常人作对照。12名患者鼻出血时应用鼻内镜下双极电凝止血术43例（次）。结果所有患者均出现ALK1基因的错义突变c．1010T〉C（p．L337P）,对照组未发现此突变。采用鼻内镜下双极电凝止血术,有效止血,且复发时间较治疗前明显延长,复发频率延长至1个月～1年以上,2例两年无出血症状。结论ALK1基因的错义突变c．1010T〉C（p．L337P）是导致该家系临床表型的主要原因,鼻内镜下双极电凝止血术处理此类病变效果良好。     

遗传性出血性毛细血管扩张症Smad4基因的筛查

目的检查遗传性出血性毛细血管扩张症(HHT)患者中Smad4基因突变及听力情况。方法根据2000年Shovlin提出的临床诊断标准,对7例ENG和ACVRL1基因筛查均阴性的HHT患者进行Smad4基因筛查和听力检查。结果 7例患者smad4基因测序均未发现突变位点,2例患者除严重鼻出血外还伴肝脏血管畸形,7例患者均无听力障碍、胃肠出血者及肠息肉患者。结论 Smad4基因与HHT、耳聋的相关性值得进一步研究,为疾病诊疗提供新的思路。     

两个耳聋家系TMPRSS3基因突变分析及产前诊断

目的对两个耳聋家系进行遗传性耳聋基因突变检测,为家系遗传咨询与产前诊断提供参考。方法2018年3—12月,郑州大学第一附属医院遗传与产前诊断中心应用二代测序技术对两个家系患儿进行耳聋基因检测(包含168个已知致病基因,包括核基因、相关线粒体区域及miRNA),并对可疑基因在患儿及家系成员中进行Sanger双向测序验证,确定致病突变后,对两个家系的高危胎儿进行产前诊断。结果家系1患儿检测到TMPRSS3基因c.432delA和c.617-2_617-1insTC复合杂合突变,家系2患儿检测到TMPRSS3基因c.271C>T(p.R91X)和c.147dupT复合杂合突变,两家系患儿父母均为携带者。产前诊断结果显示两家系胎儿均只携带1个杂合突变。随访至2019年8月,两家系二胎分别为15月龄和13月龄,听力未见异常。结论TMPRSS3基因突变可能是两个耳聋家系的致病基因,用二代测序技术可以高效、经济准确地对遗传性耳聋患者进行基因诊断,为家系遗传咨询和产前诊断提供参考。     

一个非综合征型聋家系的遗传学分析及产前诊断

目的 对一个遗传性聋家系进行致病基因鉴定、遗传咨询和产前诊断。方法 运用目标区域捕获测序检测一个非综合征型聋家系遗传学病因，对检出的致病突变进行Sanger测序验证，结合STR检测技术对该家系行产前诊断。结果 一个家系两代人(Ⅰ:2、Ⅱ:2、Ⅱ:3)均为遗传性聋，但病因不同，先证者(Ⅰ:2)是SLC26A4基因C.919-2A>G纯合突变导致，两个异卵双胞胎女儿(Ⅱ:2、Ⅱ:3)耳聋病因是MYO15A基因c.5062＿5063delCT/c.7396-1G>A复合杂合突变。先证者孕期胎儿产前诊断结果显示耳聋风险低，出生后复查与产前诊断一致，并顺利通过新生儿听力筛查。结论 本研究明确了该非综合征型聋家系的基因型，首次明确了MYO15A基因c.5062＿5063delCT突变为致病性变异，拓展了MYO15A基因致病突变谱。     

耳聋基因panel在耳聋基因诊断中的临床应用

目的探讨耳聋基因panel技术在耳聋患者基因诊断中的应用。方法40例耳聋患者首先采用荧光定量PCR结合Sanger测序法检测4个常见耳聋基因的25个位点突变,初检结果单杂合致病突变者行耳聋基因单基因测序或耳聋基因panel检测;初检结果未发现耳聋基因致病性突变者直接行耳聋基因panel检测。16例患者行父母耳聋基因溯源验证。结果40例患者中,耳聋基因筛查检出GJB2基因纯合或复合杂合突变8例、单杂合突变2例,SLC26A4基因纯合突变1例、单杂合突变2例。4例单杂合突变检出者接受进一步的耳聋单基因或耳聋基因panel测序,其中2例分别检出GJB2基因c.235delC/c.610delC及c.235delC/c.109G>A复合杂合突变,2例检出SLC26A4基因c.919-2A>G/c.1548_1549insC复合杂合突变。27例初检结果阴性患者接受了进一步的耳聋基因panel检测,检出GJB2基因c.109G>A纯合突变4例和c.571T>C/c.G109A复合杂合突变1例,MYO7A基因c.397dupC/c.3484A>T复合杂合突变1例,MYO15A基因c.4779+2T>C/c.5008-2A>G复合杂合突变1例,ACTG1基因c.118C>T单杂合突变1例,CDH23基因c.1765G>A/c.6504T>A及c.6049G>A/c.7225-1G>A复合杂合突变各1例。在16例行父母溯源的耳聋患者中,15例患者耳聋基因突变分别遗传自其父母。结论对于耳聋基因热点突变检测结果阴性的耳聋患者,应用耳聋基因panel能有效提高遗传性致病基因检出效率,为其遗传学诊断和临床治疗提供依据。     

GJB2基因在遗传性聋中的检测

目的对遗传性聋家系进行GJB2基因突变检测,为该病的基因诊断提供依据。方法采用PCR直接测序法对20个非综合征型遗传性聋家系的先证者(均为耳聋患者)进行GJB2基因的突变检测。结果发现了三种碱基改变:109G>A、79G>A和341G>A。109G>A是已报道的具有争议的致病突变,本实验在两个隐性遗传性聋家系的先证者中检测到109G>A纯合突变,且与耳聋共分离。79G>A和341G>A是已报道的多态。结论本研究发现了具有争议的致病突变109G>A的纯合突变,极可能导致隐性遗传性聋。     

A Decrease in Maxillary Sinus Pressure,as Seen in Upper Airway Allergy or Infection,Results in an Increase in Upper Airway Nitric Oxide Levels

The paranasal sinuses are connected to the nasal cavity via small osties. Ostial occlusion, caused by mucosal swelling, will result in a slowly increasing negative pressure inside the sinus cavity. In parallel, the oxygen content in the sinus will decrease, resulting in the development of relative hypoxia. Hypoxia is a powerful inducer of nitric oxide (NO) synthase, and inducible NO synthase has been shown to be present in considerable amounts in the upper airways, including the sinuses. The present study was designed to investigate whether a reduction in sinus pressure would affect upper airway NO production. Thirteen healthy volunteers were investigated. A pressure chamber was used to lower the ambient pressure to-4.9 kPa. NO was sampled from one nostril or via a drainage tube inserted into the maxillary sinus before, during and after the hypobaric exposure. When the pressure was decreased, NO levels increased from 256 &#45 15 to 316 &#45 19 ppb ( n =13, p <0.001). The NO levels remained elevated (282 &#45 21 ppb; p <0.05) when measurements were repeated 20 min after leaving the chamber. The nasal airway resistance (V2 tot ) also increased as a result of the chamber session (from 16 &#45 2° before to 21 &#45 3° after; p <0.05). An increase in NO levels was also found when the experiments were repeated with NO sampled directly from the maxillary sinus (225 &#45 6 before and 265 &#45 9 ppb after; n =6, p <0.001). For control purposes the nasal analyses were repeated again, this time under hyperbaric conditions (+4.9 kPa). This resulted in a slight decrease in the NO levels (from 273 &#45 22 to 241 &#45 17 ppb; n =10, p <0.001), but there was no change in the nasal airway resistance. We conclude that a reduction in sinus pressure, as seen in upper airway allergy or infection, may result in an increase in upper airway NO production.     

One‐stop neck lump clinic: phase 2 of audit. How are we doing?

One‐stop neck lump clinic: phase 2 of audit. How are we doing? Regular monitoring and audit of a service are integral to ensuring maintenance of efficiency and standards. This is particularly important where the quality of the service is operator dependent, as is the case in the clinical diagnosis of neck lumps and fine needle aspiration cytology. The one‐stop neck lump clinic has now been running in the department for more than 20 months. A previous article described the results of the first phase audit carried out at 6 months and had identified a waiting time to be seen that was longer than that recommended by the British Association of Otorhinolaryngologists, Head and Neck Surgeons. Measures were implemented to reduce this waiting time and a second audit was carried out after another 10 months with the aims of assessing if modification of the means of referral reduces waiting time and if the outcomes of clinical performance in phase 1 could be maintained or improved. We discuss the results of phase 2 in the audit spiral.     

A case of cortical deafness and anarthria

The nasal epithelium protects the underlying tissue from damage. Epithelial cell growth is controlled by epidermal growth factor (EGF) and is possibly affected by toxic proteins, e.g. eosinophil cationic protein (ECP). The aims of this study were to examine nasal fluid epithelial cell counts and their relations to EGF, eosinophils and ECP in 23 patients with seasonal allergic rhinitis and 20 healthy controls. Nasal fluid epithelial cell counts were lower in patients than in controls. EGF levels did not differ between patients and controls, and correlated with epithelial cell counts in controls but not in patients. Eosinophils and ECP were higher in patients than in controls, but did not correlate with epithelial cell counts. The role of growth factors, such as EGF, in regulating epithelial cells merits further study.     

A simple objective evaluation and grading for facial paralysis outcomes

Matrix metalloproteinase (MMP)-2 and -9 degrade type IV collagen, which is one of the major components of the basement membrane in normal tissue and expressed in the surroundings of the cancer nest in squamous cell carinoma. The degeneration of type IV collagen is an essential step in the metastasis to lymph nodes and distant organs. In this study, we examined MMP-2 and -9 levels of cancer tissue and serum obtained from patients with head and neck squamous cell carcinoma (HNSCC) in order to evaluate the relationship between the clinicopathologic features and MMPs. We examined the production of MMP-2 and -9 in cancer tissue homogenates of 73 patients who had HNSCC and the serum MMP levels of 16 patients with HNSCC and 8 healthy volunteers. We also studied the localization of MMP-2 in the carcinoma using an immunohistochemical approach. The concentrations of MMP-2 and -9 in the tissue homogenates and serum were measured by means of a sandwich enzyme immunoassay using a monoclonal antibody. Immunohistochemical analyses were performed with monoclonal antibody to MMP-2. The concentration of MMP-2 in the tumor tissue homogenates was unrelated to tumor size, but that in patients with lymph node metastases was significantly higher than in those without lymph node metastases. The concentration of MMP-9 was unrelated to lymph node metastasis and tumor size. The levels of both MMP-2 and -9 in serum were unrelated to lymph node metastasis. Immunohistochemistry indicated that MMP-2 was mainly expressed in cancer cells. Because MMP-2 degrades type IV collagen, the level of MMP-2 in carcinomas may be a useful indicator of the degree of invasion and metastasis.     

Bilateral versus unilateral cochlear implantation in young children

Objectives

To compare the preverbal communication skills of two groups of young implanted children: those with unilateral implantation and those with bilateral implantation.

Material and methods

The study assessed 69 children: 42 unilaterally and 27 bilaterally implanted with age at implantation less than 3 years. The preverbal skills of these children were measured before and 1 year after implantation, using Tait Video Analysis that has been found able to predict later speech outcomes in young implanted children.

Results

Before implantation there was no significant difference between the unilateral group and the bilateral group. There was still no difference at 12 months following implantation where vocal autonomy is concerned, but a strongly significant difference between the groups for vocal turn-taking and non-looking vocal turns, the bilateral group outperforming the unilateral group. Regarding gestural turn-taking and gestural autonomy, there was a strongly significant difference between the two groups at the 12 month interval, and also a difference before implantation for gestural autonomy, the unilateral group having the higher scores. Multiple regression of non-looking vocal turns revealed that 1 year following implantation, bilateral implantation contributed to 51% of the variance (p < 0.0001), after controlling for the influence of age at implantation and length of deafness which did not reach statistical significance.

Conclusions

Profoundly deaf bilaterally implanted children are significantly more likely to use vocalisation to communicate, and to use audition when interacting vocally with an adult, compared with unilaterally implanted children. These results are independent of age at implantation and length of deafness.     

Autoimmunity in Sudden Sensorineural Hearing Loss: Possible Role of Anti-endothelial Cell Autoantibodies

In order to verify whether anti-endothelial cell autoantibodies (AECAs) can be used as serological markers of inner ear vasculitis in sudden sensorineural hearing loss (SSHL), 32 patients affected by idiopathic SSHL were investigated. All patients underwent a routine general physical examination and extensive audiovestibular, microbiological and immunological investigations. Fourteen normal subjects without a history of HL, autoimmune or metabolic disease served as controls. Detection of AECAs was performed using an indirect immunofluorescence technique. AECA-positive patients were treated with methylprednisone, while AECA-negative patients were treated with a combined regimen of steroids, plasma expander and aspirin. The average hearing recovery for 5 frequencies (0.25-4 kHz) was analyzed in each subject 1 month after treatment and every 3 months thereafter; median follow-up was 12 months (range 9-18 months). A total of 15/32 patients (46.8%; 11/19 females, 4/13 males) were AECA-positive and thus differed significantly from the normal population in whom only 2/14 tested cases were positive ( p =0.03). Severe hearing loss was associated with being AECA-positive in 8/11 cases. During follow-up, 25/32 patients improved their hearing and 17 of these patients were AECA-negative. The seven cases without hearing improvement were all AECA-positive. In patients with SSHL, immune-mediated vascular damage may have a pathogenetic role and AECAs may represent a serological marker of vasculitis even if they are not inner ear-specific and even if they represent an epi-phenomenon rather than the only cause of SSHL.     

Correction of the Glosso-Larynx and Resultant Positional Changes of the Hyoid Bone and Cranium

In ankyloglossia with deviation of the epiglottis and larynx (ADEL) the tongue is located forward and as a result the epiglottis is elevated and leans towards the mouth. The larynx is also raised and curves ventrally. Various symptoms have been observed as a result of this condition. Correction of the glosso-larynx (CGL) is the operation performed to treat ADEL. The CGL procedure and the results obtained with it are reported in this paper. In addition, we studied the following six parameters using head and neck X-rays before and after CGL (the changes in these parameters as a result of CGL are shown in parentheses): ( i ) the shortest vertical length between the hyoid bone and mandible (+10.3 mm); ( ii ) the vertical length between the hyoid bone and the tangent line of C2-4 (+4.6 mm); ( iii ) the shortest length between the hyoid bone and the chin (+2.9 mm); ( iv ) the angle between the hyoid bone and the tangent line of C2-4 (+3.3°); ( v ) the length of H-M, where H is the intersection of a tangent line of C2-4 and a vertical line from the hyoid bone and M is the intersection of a tangent line of C2-4 and the mandible (+7.4 mm); and ( vi ) the width of the narrowest part of the hypopharynx (+3.0 mm). The changes in all the measured parameters after CGL were significantly different ( p <0.05).     

Carcinoma of the Nasopharynx: Analysis of Treatment Results in 91 Patients

The outcome of 91 patients (69 males, 22 females; age range 16-82 years) with nasopharyngeal carcinoma treated in our hospital between 1971 and 1999 was evaluated. Factors that appeared to influence prognosis were assessed using the Kaplan -Meier method. The cause-specific cumulative 5-year survival rate for the entire study population was 61.2%. The 1997 International Union Against Cancer classification was used for disease staging. The 5-year survival rates were as follows: 66.7% ( n ¾ 3) for Stage I; 100% ( n ¾ 2) for Stage IIA; 90.9% ( n ¾ 11) for Stage IIB; 78.8% ( n ¾ 25) for Stage III; 53.0% ( n ¾ 29) for Stage IVA; 37.5% ( n ¾ 16) for Stage IVB; and 20.0% ( n ¾ 5) for Stage IVC. The disease-free cumulative 3-year survival rates of the patients classified based on initial therapy were as follows: radiation alone, 50.0% ( n ¾ 28); combined radiotherapy and chemotherapy that included an undefined anti-cancer drug, 67.2% ( n ¾ 39); combined radiotherapy and chemotherapy that included carboplatin (CBDCA), 92.3% ( n ¾ 19). These results showed a statistically significant difference ( p ¾ 0.043; log-rank test). Stage IVC patients were excluded from the analysis. We conclude that combined therapy, including chemotherapy with CBDCA, is necessary for the treatment of nasopharyngeal carcinoma. In terms of radiation therapy, a field covering the bilateral cervical regions seemed to produce favorable results, even if cervical node metastasis was not confirmed by palpation at the first hospital visit.     

Treatment Effects in Patients with Squamous Cell Carcinoma of the Oral Cavity

A total of 221 patients (155 males, 66 females; stage I, n ¾ 55; stage II, n ¾ 58; stage III, n ¾ 57; stage IV, n ¾ 51) with squamous cell carcinoma of the oral cavity were studied. Tumor localization was as follows: cancer of the tongue, n ¾ 161; cancer of the oral floor, n =28; cancer of the hard palate, n ¾ 12; cancer of the buccal mucosa, n ¾ 11; and cancer of the gingiva, n ¾ 9. In order to compare the effect of different treatments, three major treatment groups were defined, namely a surgery group, a radiotherapy group and a combination treatment group. Five-year cumulative survival rates showed significant differences between stage classifications (stage I=91%, stage II=73%, stage III=63%, stage IV=47%; p <0.01) but not between tumor sites. The 5-year cumulative survival rate was highest for oral floor cancer (80%). In the early-cancer group, the 5-year cumulative survival rate for the surgery group (92%) was significantly higher ( p <0.05) than those for both the radiation (69%) and combination (71%) groups. In the advanced-cancer group, the 5-year cumulative survival rate for the surgery group (74%) was significantly higher ( p <0.05) than those for both the radiation (37%) and combination (51%) groups. No significant difference in regional control rates was observed between the treatment groups. Five-year regional control rates were 86% for cervical untreated patients with T1N0 tumors and 60% for cervical untreated patients with T2N0 tumors. Fourteen N0 cases were treated with neck dissection. Cervical metastasis was found pathologically in 2/14 (14%) of these cases. The 5-year survival rate for patients with cervical recurrences after primary tumor resection was 70% ( n ¾ 15). In contrast, the 5-year survival rate for patients with both primary tumor resection and neck dissection was 74% ( n ¾ 14) but no significant difference was observed between these 2 groups.rate .     

Otolaryngologists may not be doing enough to diagnose pediatric eosinophilic esophagitis

Objective

To systematically evaluate the diagnosis of eosinophilic esophagitis (EE).

Methods

A retrospective review of 657 patients seen at the EE center of a tertiary care children's hospital between 1994 and 2007 was performed. Charts were reviewed for the 144 patients who were also seen by the otolaryngology service.

Results

One hundred forty-four patients received 193 otolaryngology-related diagnoses. Eustachian tube dysfunction (27.5%) and sleep disordered breathing (24.9%) were the most common, followed by dysphagia (13.0%), rhinosinusitis/nasal congestion (9.3%) and airway stenosis (5.2%). Seventy-nine patients (54.9%) had a pre-existing diagnosis of EE at the time of their otolaryngology consultation. Twenty-one patients (14.6%) were referred to the gastroenterology service for evaluation for EE. Forty-four patients (30.5%) remained undiagnosed. Twenty-five of these patients presented with dysphagia, 16 of whom were not previously diagnosed with EE; only 4 of these 16 patients were referred for evaluation for EE. In one case, a child with moderate sized tonsils underwent adenotonsillectomy for dysphagia and failure to thrive; this patient was diagnosed with EE 1 month post-operatively.

Conclusions

Twenty percent of patients with EE may require care by an otolaryngologist for a myriad of complaints. Even experienced pediatric otolaryngologists may not recognize this condition. Otolaryngologists should consider EE in patients presenting with dysphagia. A careful gastroenterology review of symptoms may also allow otolaryngologists to identify EE in patients with allergy mediated nasal complaints, or laryngeal/airway disorders.