自身免疫性感音神经性聋血清抗膜迷路蛋白抗体的检测

采用改良免疫转印法分析了拟诊为自身免疫性感音神经性听力减退患者休内抗膜迷路蛋白抗体，并将该法与常规法比较，发现用该法在患者血清中最多可检出５条强阳性（含６８０００），阳性率为７２％（１８／２５），显著高于常规法（４０％）。还在部分患者体内检出了抗肺炎克雷白杆菌膜蛋白抗体（７／９）。     

酶联免疫吸附试验检测抗耳蜗及蜗神经组织相关抗体的应用

自制牛耳蜗抗原（ＣＣＡｇ）及牛蜗神经抗原（ＣＣＮＡｇ）采用酶联免疫吸附试验（ＥＬＩＳＡ）对５２例（８９侧）不明原因感音神经性聋患者及对照组２０例无耳疾和免疫性疾病的患者进行了检测，结果抗ＣＣＡｇ抗体水平对照组为０．３８１１±０．０７４３，（吸光度Ａ值，ｘ±ｓ，下同），实验组为０．９４２１±０．２４０８，两组差异有高度显著性（Ｐ＜０．０１），以对照组抗ＣＣＡｇ抗体Ａ值的ｘ＋２ｓ判断，实验组２４例抗ＣＣＡＧ抗体阳性，阳性率为４６％。抗ＣＣＮＡｇ抗体水平对照组为０．２２５４±０．０３８２，实验组为０．２３３１±０．０６５６，两组差异无显著性，以ｘ＋２ｓ判断，实验组６例抗ＣＣＮＡｇ抗体阳性，阳性率为１１．５％。认为ＥＬＩＳＡ检测法程序简便，可动态观察患者抗体水平，并能定量分析，配合免疫组织化学方法或免疫转印法可用于内耳自身免疫病的诊断。用ＥＬＩＳＡ法检测到抗ＣＣＮＡｇ抗体，证实了国内外学者的推断，即内耳自身免疫病存在蜗后起源的可能性。     

抗内耳抗体与突发性聋

目的研究突发性聋患者血清中是否存在抗内耳抗体,探讨内耳抗体与突发性聋的相关性。方法经病史询问、耳科检查和听力学检查,由北京同仁医院耳科门诊诊断的突发性聋患者为研究对象。采用豚鼠内耳石蜡切片作为抗原,用间接免疫荧光法检测突发性聋患者体内的抗内耳自身抗体。结果74.0%（37/50）的突聋患者存在各种抗内耳组织抗体,12.8%（9/70）的对照组个体存在抗内耳抗体;经统计学分析P〈0.01,有显著性差异。44.0%（21/50）的突聋患者存在抗内耳血管内皮抗体和抗内耳血管壁抗体,突聋患者还存在其他类型的抗内耳组织抗体。结论突发性聋患者血清中存在各种类型的抗内耳组织抗体。抗内耳抗体与突发性聋的关系需要进一步探讨。     

二黄冲剂对鼻咽癌癌前病变患者EBV-VCA/IgA抗体阳性治疗作用观察

采用随机，单盲法对鼻咽癌癌前病变EBV－VCA/IgA抗体滴度＞1：10的患者62例进行治疗。结果：二黄冲剂治疗组（32例）总显效率75％（24／32），阴转率43．7％（14／32），鼻咽清毒剂组（30例）总显效率13·3％（4／3O），阴转率10％（3／30），两组统计学处理差异显著。提示二黄冲剂能显著降低或阴转血清EBV－VCA／IgA抗体滴度，可产生较好的鼻咽癌预防效应。     

成年人慢性鼻-鼻窦炎鼻内镜术后的细菌学研究

目的探讨成年人慢性鼻-鼻窦炎（chronicrhinosinusitis，CRS）患者鼻内镜手术（endoscopicsinussurgery，ESS）后的细菌分布和耐药性特征。方法实验组来自87例CRS患者ESS术中中鼻道、上颌窦、筛窦和术后1、3、6个月复诊时的术腔分泌物；对照组为30例鼻中隔偏曲（无鼻炎、鼻窦炎）患者的中鼻道分泌物。所取标本分别作细菌培养、药物敏感实验及B内酰胺酶菌株检测。结果464份标本共检出细菌645株26种，总细菌检出阳性率78．9％（366／464）。其中革兰阴性菌（Gramnegativebacteria，GNB）占51．2％（330／645）；革兰阳性菌（Grampostivebacteria，GPB）占48．8％（315／645）；73．6％（64／87）的患者为混合菌生长。需氧菌为主占95．3％（615／645），厌氧菌仅占4．7％（30／645）。CRS患者术后GNB检出阳性率较术前明显增加，以产气肠杆菌、铜绿假单胞菌和流感嗜血杆菌最为常见；术后1个月、3个月组厌氧菌检出阳性率为3．4％（3／87）和2、3％（2／87），较术前9．2％（8／87）明显降低，而迁移不愈组反而升高，占15．4％（2／13）。术后多重耐药（mutipledrugresistance，MDR）菌株检出阳性率较术前明显增加。迁移不愈组B内酰胺酶检出率为30．8％（4／13），其中以铜绿假单胞菌最为常见；而术前阳性检出率为19．5％（17／87），以金黄色葡萄球菌为主，两组间差异有统计学意义（X2=4．85，P〈0．05）。痊愈组与对照组间B内酰胺酶阳性率和菌种分布的差异无统计学意义。结论GNB是CRS患者术后不容忽视的重要条件致病菌或致非条件病菌，CRS术后迁延不愈与MDR-GNB的优势生长有一定的关系。术后痊愈患者鼻腔和鼻窦的细菌微生态逐步恢复平衡。     

豚鼠膜迷路蛋白组成分析及免疫转印法检测感音神经性聋患者抗68kD抗体

比较了多种条件提取的豚鼠膜迷路蛋白，发现用十二烷基磺酸钠（ＳＤＳ）或ＴｒｉｔｏｎＸ－１００提取效率高，冻融法较超声粉碎法更可取。成功地建立了免疫转印（Ｗｅｓｔｅｒｎｂｌｏｔ）法，并初步运用于血清抗膜迷路蛋白抗体的检测。４２例不明原因感音神经性聋患者中１６例患者血清中抗６８ｋＤ抗体阳性，同时伴有ＩｇＧ、ＩｇＭ、ＩｇＡ升高。部分正常对照组血清中也存在该抗体，但反应弱，重复性差。     

耳廓假性囊肿免疫学病因的初步探讨

目的　探讨耳廓假性囊肿发病与全身免疫功能及局部免疫状态的关系。方法　采集４４ 例耳廓假性囊肿患者的囊液和外周静脉血标本，用免疫单扩散法分别检测囊液和血清中的抗体ＩｇＧ、ＩｇＡ、ＩｇＭ 和补体Ｃ３ 水平；用聚乙二醇比浊法分别检测囊液和患者血清中的免疫复合物含量；用聚合酶链反应检测囊液中巨细胞病毒ＤＮＡ；用免疫荧光技术检测囊液和血清中的抗核抗体，用间接荧光抗体检测法（ｉｎｄｉｒｅｃｔｉｍｍｎｏｆｌｕｏｒｅｓｃｅ ａｓｓａｙ，ＩＦＡ） 检查囊液和患者血清与健康小鼠耳廓冰冻组织切片的反应情况。结果　耳廓假性囊肿患者囊液中抗体ＩｇＧ、ＩｇＡ、ＩｇＭ 和补体Ｃ３ 水平均明显低于血清中者（ Ｐ＜ ０．０１），囊液和患者血清中均未检出免疫复合物，囊液中也未检测出巨细胞病毒ＤＮＡ；但在部分（３／１０） 患者囊液中发现了抗核抗体，在小鼠冰冻切片中发现了位于软骨组织上的抗原抗体复合物。结论　耳廓假性囊肿中部分患者可能是局限于耳廓的自身免疫性疾病，可能是由于自身抗体与耳廓组织发生自身免疫反应导致反应性渗出液在局部聚集形成囊肿。     

豚鼠膜迷路蛋白组成分析及免疫转印法检测感音神经…

比较了多种条件提取的豚鼠膜迷路蛋白，发现用十二烷基磺酸钠（ＳＤＳ）或Ｔｒｉｔｏｎ Ｘ－１００提取效率高，冻融法较超声粉碎法更可取。成功地建立了免疫转印（Ｗｅｓｔｅｒｎ ｂｌｏｔ）法，并初步运用于血清抗膜迷路蛋白抗体的检测。４２例不明原因感音神经性聋患者中１６例患者血清中抗６８ｋＤ抗体阳性，同时伴有ＩｇＧ、ＩｇＭ、ＩｇＡ升高。部分正常对照组血清中也存在该抗体，但反应弱，重复性差。     

抗人喉癌单克隆抗体对荷瘤裸鼠放射免疫定位显像的观察

目的 用喉癌裸鼠模型研究抗人喉癌单克隆抗体的免疫导向作用，为诊断和治疗提供依据和方向。方法 用高效碘标法标记三株混合抗人喉癌单克隆抗体（ＭｃＡｂＬＣ）经腹腔注入人喉癌裸鼠模型体内，用单光子发射计算机断层（ＥＣＴ）扫描显像并测定肿瘤、血液和组织单位质量浓集的放射性占注入总放射性的百分比（％ＩＤ／ｇ）和肿瘤与主要器官组织的放射性比值（Ｔ／ＮＴ）。结果 单克隆抗体在一定时限内能够选择性的浓集在移植瘤组织     

100例鼻咽癌患者及其家属血清中EBV－VCA－IgA抗体检测

１００例鼻咽癌患者及其家属血清中ＥＢＶ－ＶＣＡ－ＩｇＡ抗体检测赵素萍，肖健云，谭莉，陶正德鼻咽癌患者血清中可检出高滴度抗非洲淋巴细胞瘤（ＥＢＶ）相关抗体，尤以病毒壳蛋白抗原－免疫球蛋白Ａ（ＶＣＡ－ＩｇＡ）抗体在鼻咽癌患者更具有特异性，故认为ＥＢＶ感染...     

Bcl—2,Bax蛋白产物在鼻咽癌中的表达,分布及意义

目的：探讨Ｂｃｌ－２、Ｂａｘ蛋白产物在鼻咽癌组织中的表达情况。方法：应用免疫组化技术Ｓ－Ｐ法,对４２例ＮＰＣ和１０例正常鼻咽粘膜组织标本,进行了调亡相关基因Ｂｃｌ－２和Ｂａｘ蛋白产物检测。结果：（１）在正常鼻咽粘膜中,Ｂｃｌ－２蛋白主要分布在基底层细胞,而Ｂａｘ蛋白分布在基底层以上细胞;（２）在ＮＰＣ组织中,Ｂｃｌ－２和Ｂａｘ蛋白产物表达阳性率为８５．７％和８３％,且Ｂｃｌ－２蛋白表达与ＢＰＣ颈淋     

Expression of steroid receptors in acoustic neuroma

The expression of steroid receptors has been investigated in an attempt to clarify the role of steroid hormones in the proliferation and progression of acoustic neuromas. Specimens of tumours taken during translabyrinthine surgery were tested for cytosolic (c) and nuclear (n) steroid receptors. Oestrogen and progesterone receptor levels were evaluated by enzymatic immuno-assay, while androgen receptor binding levels were detected by dextran-coated charcoal method in a single-step determination. In some cases, the six point Scatchard analysis of cytosolic and nuclear androgen receptor was also performed. Threshold values were: 3 fmol/mg of proteins for cytosolic steroid receptors and 20 fmol/mg DNA for nuclear steroids, which corresponded to approximate median values of cytosolic and nuclear oestrogen and progesterone, respectively. Oestrogen and progesterone appeared to be localized more frequently in the nuclei rather than in the cytosol (70% oestrogen and progesterone positivity in the nuclei; 30% oestrogen, 40% progesterone positivity in the cytosol), while androgen receptors were preferentially localized in the cytosol (80% positivity in the cytosol; 40% positivity in the nuclei). A negative non-linear correlation between cytosolic oestrogen and cytosolic androgen receptors was found. There was a direct linear correlation between cytosolic oestrogen and nuclear oestrogen levels. A strict correlation between nuclear oestrogen and nuclear progesterone incidence was shown. Preliminary analysis of clinical data and biochemical parameters showed that cytosolic progesterone levels inversely correlated with tumour size.     

bcl-2、P 5 3基因及Epstein-Barr病毒在鼻咽癌中的表达

目的探讨鼻咽癌组织中p53、bcl-2、EBV表达情况及与主要临床指标间关系.方法免疫组化检测p53、bcl-2蛋白,多聚酶链反应(PCR)检测EBV.结果慢性炎性组织中p53、bcl-2及EBV阳性率分别为7.7%、23.1%和7.7%,明显低于鼻咽癌组织中的阳性率(分别为75.9%、88.9%和87.0%)(P<0.01).三者在鼻咽癌组织中共同阳性率为72.2%,p53、bcl-2明显正相关(r＝0.78,P<0.01).鼻咽癌伴颈淋巴结转移组p53阳性率(90.0%)明显高于无颈淋巴结转移组(58.3%)(P<0.01),临床Ⅱ、Ⅲ和Ⅳ期p53阳性率(分别为92.9%、78.9%和100%)均明显高于临床I期(38.5%)(P<0.01),且表达强度有增加的趋势.结论p53、bcl-2及EBV可作为鼻咽癌早期诊断的辅助指标之一,p53阳性的鼻咽癌患者较易发生颈淋巴结转移,随着鼻咽癌病程的发展,p53表达率及表达强度均增加,三者在鼻咽癌发生发展中起着重要作用.     

Significance of Fas protein in squamous cell carcinoma of the larynx

The purpose was to elucidate the clinical significance of Fas protein in laryngeal squamous cell carcinoma (LSSC). Medical records of 30 patients who were operated for LSCC between 1991 and 1997 were reviewed, and histopathological and immunohistochemical studies of the archival tumor tissues were performed. Conventional histopathology included tumor grading, mitotic counting (MC) and status of the cervical lymph nodes. The monoclonal antibodies to Fas (DAKO M 3554) and nm23 (DAKO A0096 Lot 035) proteins were used for immunohistochemical staining. Streptavidin-Biotin, horseradish method was used. Positive staining for both Fas and nm23 was obtained in 27 (90%) tumor samples. There was no correlation between neck metastasis and Fas positivity (r = 0.126, p = 0.505 and chi2 = 0.062, p = 0.996, respectively). Fas positivity did not correlate to grade (r = 0.282, p = 0.130), tumor size or T (r = 0.0457, p = 0.810, chi2 = 0.0.018, p = 0.999) or laryngeal site of involvement (chi2 = 0.0937, p = 0.992). However, a correlation was found between Fas and MC (r = 0.482. p = 0.007). There was also a correlation between Fas and nm23 positivity (r = 0.614, p < 0.0001). Fas protein expression has no prognostic value in LSCC, because the Fas values do not correlate with grade, stage or nodal status of neck. However, 90% of LSCCs expressed Fas protein. Understanding the functional status of the Fas receptor proteins may lead to development of new therapeutic options for LSSC in the future.     

Hörscreening mit automatisch bewerteten TEOAE und einem neuen Verfahren automatisch bewerteter FAEP Optimierung und Feldversuch

BACKGROUND AND OBJECTIVE: To make a rational decision as to which screening test might be adequate as a universal newborn hearing screening, different methods have to be tested under "real-life" conditions. In addition, a good reference is required as "golden standard." PATIENTS/METHODS: In the study presented here, an ABR with a novel algorithm for threshold-estimation was optimized and compared to TEOAE (Echoscreen) and DPOAE (GSI 60) in a three-step protocol using a standard click-evoked ABR (Evoselect) as reference of sensitivity and specificity. RESULTS: 26 ears were found to be hearing-impaired. All of them were also detected by each of the screening method (sensitivity 100%). Specificity in the final "step 2" of the study was around 87.7/92.3% for Echoscreen, 82.4/84.4% for DPOAE and 82.4/89.1 for Evoflash (left/right, respectively). Differences were not at all significant (X2-test, p > > 0.05). In summary, results were somewhat inferior to those found by other authors, probably due to more difficult conditions in the "real-life" setting. CONCLUSIONS: A higher rate of false-alarm shows that a two-step screening is necessary. Therefore, increased resources for diagnostic procedures are required.     

Expression and clinical significance of epidermal growth factor receptor and type 1 insulin-like growth factor receptor in nasopharyngeal carcinoma

OBJECTIVES: Epidermal growth factor (EGF) and type 1 insulin-like growth factor (IGF-1) receptors play an important role in the growth and apoptosis of nasopharyngeal carcinoma (NPC). They were separately found to be associated with prognosis in patients with NPC. To date, their expression correlation and clinicopathologic significance have never been specifically addressed in NPC. METHODS: Seventy-five patients with NPC and 21 noncancerous nasopharyngeal epithelial samples were accrued between 1998 and 2006 in a single hospital. The expressions of EGF and IGF-1 receptors were detected by immunohistochemical staining in the 75 NPC samples and the 21 noncancerous samples. Furthermore, the messenger RNA and protein expressions were assessed by real-time quantitative polymerase chain reaction and the Western blot technique, respectively, in NPC cell lines and normal nasopharyngeal epithelial cells. RESULTS: The 5-year survival rates, assessed by the Kaplan-Meier method, were 71.4% and 66.6% in the EGF and IGF-1 receptor protein-negative groups, respectively, whereas they were only 28.6% and 33.3% in the receptor protein-positive groups. The levels of these two proteins significantly correlated with each other, and the overexpression rates of EGF and IGF-1 receptors were 65.3% and 56% in nasopharyngeal samples, respectively. Furthermore, both protein expressions were significantly higher in NPC patients with cervical lymph node or distant metastasis than in NPC patients without lymph node or distant metastasis. Recurrence more often appears in cases positive for both proteins than in cases negative for both proteins. The expression levels of the receptor messenger RNA and proteins were higher in several NPC cell lines than in normal nasopharyngeal epithelial cells. CONCLUSIONS: These findings demonstrate that both receptor proteins may play an important role in the invasion, metastasis, and recurrence of NPC. Both receptors are valuable markers for assessing the prognosis of NPC. Their expression at such high frequencies provides the basis of combined targeted therapy with specific pharmacologic inhibitors to enhance the effects of radiotherapy and chemotherapy.     

Serum amyloid A, properdin, complement 3, and toll-like receptors are expressed locally in human sinonasal tissue

BACKGROUND: There is a growing appreciation of the role that nasal mucosa plays in innate immunity. In this study, the expression of pattern recognition receptors known as toll-like receptors (TLRs) and the effector molecules complement factor 3 (C3), properdin, and serum amyloid A (SAA) were examined in human sinonasal mucosa obtained from control subjects and patients with chronic rhinosinusitis (CRS). METHODS: Sinonasal mucosal specimens were obtained from 20 patients with CRS and 5 control subjects. Messenger RNA (mRNA) was isolated and tested using Taqman real-time polymerase chain reaction with primer and probe sets for C3, complement factor P, and SAA. Standard polymerase chain reaction was performed for the 10 known TLRs. Immunohistochemistry was performed on the microscopic sections using antibodies against C3. RESULTS: Analysis of the sinonasal sample mRNA revealed expression of all 10 TLRs in both CRS samples and in control specimens. Expression of the three effector proteins was detected also, with the levels of mRNA for C3 generally greater than SAA and properdin in CRS patients. No significant differences were found in TLR or innate immune protein expression in normal controls. Immunohistochemical analysis of sinonasal mucosal specimens established C3 staining ranging from 20 to 85% of the epithelium present. CONCLUSION: These studies indicate that sinonasal mucosa expresses genes involved in innate immunity including the TLRs and proteins involved in complement activation. We hypothesize that local production of complement and acute phase proteins by airway epithelium on stimulation of innate immune receptors may play an important role in host defense in the airway and, potentially, in the pathogenesis of CRS.     

Alloiococcus otitidis in acute otitis media

OBJECTIVE: The bacterium Alloiococcus otitidis has been found to be associated with otitis media with effusion (OME). When the culture method is used, its detection rate is low, whereas applying the polymerase chain reaction (PCR) yields significantly higher frequencies. This study was carried out to investigate the incidence of A. otitidis in children with acute otitis media (AOM). METHODS: Multiplex PCR was used to detect A. otitidis together with Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae in the middle ear effusions (MEEs) of 118 children with AOM. The clinical outcome of AOM and the bacterial findings of MEEs were compared. RESULTS: A. otitidis was detected in 25% (30 of 118) of the tested MEE samples. Children over 2 years of age had significantly more often A. otitidis-positive MEEs (37%; 22 of 59) than younger children (14%; 8 of 59) (chi-square test, P<0.01). There were no significant differences in the duration, clinical failures (after antibiotic treatment), or number of recurrences of AOM between the A. otitidis-positive and A. otitidis-negative children. CONCLUSIONS: A. otitidis is found from the MEEs of AOM. The present data suggest that it has no clinical significance in AOM, and it does not increase the risk of developing OME after AOM.     

Chronic Suppurative Otitis Media — A prospective study

A random selection of 120 cases of Chronic Suppurative otitis Media (CSOM) were investigated for bacterial and fungal isolates at the Department of Microbiology and Parsitology, Benghazi. The specimens were obtained from the out- patient department of Otolaryngology of 28th March Polyclinic. The incidence of CSOM in Benghazi was found to be 23.7%. A total of 145 bacterial isolates were identified and their susceptibility to various antibiotics was tested. Staphylococcus aureus was the predominant pathogen isolated (42.5%), followed by Pseudomonas aeruginosa (21.6%) and proteus species (18.33%). Fungi were isolated from 7.5% of cases, but no growth was observed in 10% of specimens collected. The majority of the bacterial isolates showed multiple drug resistance. Gentamycin still formed the drug of choice in treating the causative strains of either Gram positive or Gram negative bacteria, while tetracycline was found to be the least effective.     

MAPK、PI3K-AKT、NF-κB在小鼠过敏性鼻炎中的表达及意义

目的 通过构建小鼠过敏性鼻炎模型探讨多信号通路蛋白在过敏性鼻炎发生发展中的机制。 方法 40只昆明小鼠随机分为过敏性鼻炎组和对照组(每组20只)。分离两组小鼠头部各8只,4%多聚甲醛固定、石蜡包埋、切片、HE染色法观察小鼠鼻黏膜病理改变、免疫组化检测MAPK、NF-κB、PI3K-AKT信号通路蛋白的表达,24只小鼠鼻黏膜蛋白用于Western blotting分析。 结果 过敏性鼻炎组小鼠的抓挠、喷嚏、流鼻涕等均远多于对照组,差异具有统计学意义(P<0.05);HE染色过敏性鼻炎组黏膜出现假复层纤毛柱状上皮坏死,上皮结构重构,嗜酸性粒细胞浸润等典型鼻炎病理改变。过敏性鼻炎组三条通路关键蛋白的免疫阳性信号AOD值不同程度的高于对照组(P<0.05)。Western blotting结果显示过敏性鼻炎组与对照组相比MAPK、PI3K-AKT、NF-κB三条信号通路蛋白表达升高,差异有统计学意义(P<0.05)。 结论 在小鼠过敏性鼻炎发病过程中MAPK、NF-κB、PI3K-AKT三条信号通路蛋白的表达均升高,其激活在小鼠过敏性鼻炎过程中可能存在协同效应。