聚合酶链反庆检测角膜组织中的单疱病毒DNA

应用聚合酶链反应（ＰＣＲ）技术检测正常和病变角膜上皮等组织中的ＨＳＶ－１基因ＤＮＡ。结果提示角膜组织可能是ＨＳＶ－１另一潜伏部位。     

聚合酶链反应检测角膜组织中的单疱病毒DNA

应用聚合酶链反应（ＰＣＲ）技术检测正常和病变角膜上皮等组织中的ＮＳＶ─１基因ＤＮＡ，结果表明，８例正常角膜上皮中１例检测到ＨＳＶ─１ＤＮＡ，临床诊断为活动期ＨＳＫ的３０例中１６例ＨＳＶ─１ＤＮＡ阳性，其中树枝状角膜炎７１％阳性，３例ＨＳＫ静止期白斑中２例检测到ＨＳＶ─１ＤＮＡ，９例临床诊断非病毒性角膜炎中２例ＨＳＶ─１阳性，表明ＰＣＲ是一快速、敏感、特异的方法，对角膜炎有早期诊断和鉴别诊断价值的技术，还提示角膜组织可能是ＨＳＶ─１另一潜伏部位。     

用多聚酶链反应检测泪液中单疱病毒DNA

本文应用ＰＣＲ技术，以单纯疱疹病毒（ＨＳＶ）多聚酶基因引物检测２０例正常人泪液，结果全部为阴性。检测６８例各种类型角膜炎泪液，２９例阳性结果。其中临床诊断树枝状或地图状角膜炎１２例，全部检测到ＨＳＶＤＮＡ；角膜没有典型病变、临床未定性角膜炎３０例，有１３例（４３．３％）阳性结果；单纯盘状角膜炎１２例，全部为阴性。实验结果表明：应用ＰＣＲ技术能够从单疱角膜炎（ＨＳＫ）患者的泪液中检测到ＨＳＶＤＮＡ，可以作为ＨＳＫ的病原学诊断方法，尤其对于早期或没有典型病变、角膜炎不能定性者，价值更高。     

原位聚合酶链反应检测角膜组织中HSV—1型DNA

为研究单纯疱疹病毒Ⅰ型（ＨＶＳ－Ⅰ）在角膜内的潜伏感染情况。采用原位聚合酶链反应（ＩＳＰＣＲ）定位检测２１例角膜标本中的ＨＳＶ－Ⅰ，其中７例活动期ＨＳＫ，１３例静止期ＨＳＫ，１例圆锥角膜。结论：在ＨＳＫ活动期及静止期中，角膜各层细胞内均有ＨＳＶ－Ⅰ的存在，极可能整合于细胞核内     

聚合酶链反应检测角膜内皮炎房水中单纯疱疹病毒Ⅰ型DNA及带状疱疹病毒DNA

目的 评价聚合酶链反应技术（PCR）对角膜内皮炎的诊断价值，并以此解释病因，指导临床治疗。方法对临床诊断为角膜内皮炎的患者12例（12眼）及临床诊断为年龄相关性白内障的患者15例（15眼）分别用PCR方法进行房水中单纯疱疹病毒Ⅰ型（HSV-Ⅰ）DNA及带状疱疹病毒（VZV）DNA的检测，结果采用分类变量资料两样本率比较的四格表确切概率比较。结果共扩增角膜内皮炎患者12例（12眼），HSV-Ⅰ阳性者5例（5眼），阳性率为41．67％，共扩增对照组15例（15眼），HSV-Ⅰ阳性者0例，阳性率为0％，二者差异有显著性。结论PCR技术检测角膜内皮炎患者房水中的HSV-Ⅰ，可在分子水平上建立一种快速准确的诊断方法，为临床治疗提供可靠的依据。     

单纯疱疹病毒性角膜炎角膜组织病毒潜伏感染的PCR检测分析

 目的  探讨通过穿透性角膜移植获取的单纯疱疹病毒性角膜炎病变角膜组织中1型单纯疱疹病毒（HSV-1）DNA的表达情况及意义。设计 实验研究。研究对象  2010年5-12月北京同仁医院因病毒性角膜炎角膜白斑行穿透性角膜移植术后角膜标本20例,圆锥角膜、大泡性角膜病变和角膜营养不良等非感染性角膜病变的角膜标本20例。方法  对角膜组织标本中HSV-1 DNA进行聚合酶链反应（PCR）检测。 主要指标  HSV-1 DNA的阳性率。结果  单纯疱疹病毒性角膜炎静止期患者角膜组织中12/20例（60%）检出HSV-1 DNA,非感染性角膜组织中6/20例（30%）检出HSV-1 DNA（χ2=3.64,P=0.057）。结论  单纯疱疹病毒性角膜炎静止期角膜组织多数表达HSV-1 DNA,角膜内潜伏病毒是引起单纯疱疹病毒性角膜炎的可能原因,正常人角膜也可能有HSV-1的DNA存在。（眼科,2013,22：45-48）     

格子状角膜营养不良患者BIGH3基因突变的研究

目的探讨中国格子状角膜营养不良(LCD)患者基因突变类型。方法对2002年7至11月来我院就诊的8例不伴有全身淀粉样物质沉积的LCD患者,自静脉血提取全血白细胞DNA,应用聚合酶链反应(PCR)技术,扩增BIGH3基因目的片段,并对其进行DNA直接测序;同时行裂隙灯显微镜检查并照裂隙灯显微镜外眼像。收集32名正常人静脉血样进行同样检测,作为对照。结果3例检出BIGH3基因第4外显子的R124C突变(417位点碱基C→T),呈杂合子,临床表现为典型的LCDI型;另外5例检出第14外显子的H626R突变(1924位点碱基A→G),亦为杂合子,该型临床表现介于LCDⅠ型与LCDⅢ或ⅢA型之间,为一中间类型。结论中国LCD患者中既存在引起LCDⅠ型的R124C突变,也存在H626R突变。因此,H626R突变并非是英国和法国特有的类型,也可为亚洲患者的一种基因突变类型。     

鼻咽癌放疗后化疗敏感性的变化

目的检测鼻咽癌CNE1细胞x射线照射前后多药耐药基因（mdr1基因）及其编码产物P糖蛋白（P—glycoprotein，p-gP）是否表达，以及化疗药物敏感性的变化。方法利用RT—PCR、Western blotting和流式细胞仪（FCM）检测CNE1细胞x射线照射前后的mdr1基因和P—gp的表达及对柔红霉素（daunorubicin，DNR）的外排功能。结果鼻咽癌CNE1细胞射线照射前mdr1基因、P．gp不表达；射线照射后较长时间内mdr1基因、P—gp均长时间表达。CNE1细胞射线照射后对DNR的蓄积作用略有降低。结论鼻咽癌CNE1细胞x射线照射后化疗敏感性降低。     

鼻咽癌患者调强放射治疗后鼻窦炎的临床特征分析

目的 探讨鼻咽癌(NPC)患者调强适形放射治疗(简称调强放疗)后鼻窦炎的临床特点及发展规律,为更好地进行早期临床干预提供依据。方法 对已行鼻咽部病理活检确诊为NPC拟行放疗的患者进行研究。比较不同时间节点(放疗前、放疗结束时、结束后3个月、结束后6个月、结束后12个月)鼻窦炎各项评分(鼻窦炎症状评分,鼻内镜评分,CT/MRI评分),分析各项评分与T分期、鼻腔鼻窦解剖异常的相关性。结果 NPC放疗后鼻窦炎症状评分、鼻内镜评分、CT/MRI评分在不同时间节点之间的差异均有统计学意义(χ2值分别为19.28、22.33、67.73,P值均<0.01),各项评分在放疗结束后6个月达到峰值。不同时间节点的鼻窦炎症状评分、CT/MRI评分(除放疗结束后6个月外)均与T分期呈正相关性(P<0.05)。在不同时间节点,各项评分与鼻腔鼻窦解剖异常均无明确相关性。结论 NPC患者调强放疗后鼻窦炎有其自身的临床特点及发展规律,放疗后6个月炎症程度达峰值,不同时间节点各项评分多数与T分期相关,而与鼻腔鼻窦解剖异常无明显相关性。     

上海地区鼻咽癌患者血清EB病毒抗体检测结果分析

目的 评估EB病毒的Rta蛋白抗体(Rta/IgG)、核抗原抗体(EBNA1/IgA)和病毒衣壳抗原抗体(VCA/IgA)的检测对上海地区鼻咽癌患者的临床诊断价值.方法 收集206例病理证实未经治疗的鼻咽癌患者和249例健康成人的血清,均采用酶联免疫吸附试验法定量检测血清中的Rta/IgG、EBNA1/IgA、VCA/IgA抗体水平.结果 206例鼻咽癌患者的EB病毒抗体Rta/IgG、EBNA1/IgA、VCA/IgA的敏感度分别为64.08％、88.35％、86.89％;特异度分别为92.62％、92.92％、95.69％;3种抗体同时阳性的模式在鼻咽癌患者中出现的概率最高(55.83％),而在健康对照组中无一例出现.结论 Rta/IgG、EBNA1/IgA、VCA/IgA的联合检测对上海地区鼻咽癌患者的诊断具有参考价值.(中国眼耳鼻喉科杂志,2012,12:40-41,46)     

鼻咽癌放疗后大出血15例诊疗体会△

目的 探讨鼻咽癌(NPC)患者放疗后大出血的原因以及可行的治疗和挽救措施。方法 回顾分析2012年1月~2018年10月就诊于我科的15例NPC放疗后出血患者的临床资料。所有患者均予以心电监护、补液扩容、鼻腔填塞等处理。8例予输血,4例行气管插管。9例行数字减影血管造影术(DSA),其中5例提示颈内动脉假性动脉瘤(PSA)形成,1例行岩骨段覆膜支架置入,1例在外院行颈内动脉闭塞术,其余3例未行治疗。结果 15例患者中死亡5例,自动出院4例(2例深度昏迷),好转出院6例。行覆膜支架置入者术后好转出院,随访提示术后2个月死亡。行颈内动脉闭塞术者术后肢体偏瘫,经康复治疗后改善。未行治疗者均死亡。4例患者行颌内动脉栓塞术,3例好转出院,1例死亡。结论 NPC放疗后大出血病情凶险,预后较差。出血部位可为颌内动脉出血及颈内动脉PSA形成。鼻腔填塞是止血的首选措施,鼻咽部填塞及DSA为可选方法。     

鼻咽癌长期放射治疗后双眼缺血综合征1例

目的：报道1例因接受放射治疗后出现双侧颈总动脉完全闭塞和双眼缺血综合征(ocular ischemic syndrome,OIS)的患者。 方法：病例报告。1例57岁男性患者主诉左眼一过性黑矇伴有眼部和眶周疼痛6mo,诊断为双侧OIS,依次给予双侧全视网膜光凝(panretinal photocoagulation,PRP)、白内障手术、颈动脉内膜剥脱术和Ahmed青光眼阀植入术。 结果：经过PRP和白内障超声乳化手术治疗,双侧眼部病情稳定。但患者行双侧颈动脉内膜剥脱术后出现眼压升高,视力降至指数。 结论：鼻咽癌放射治疗后出现的双侧颈总动脉完全闭塞和双眼OIS的治疗比较困难,预后不佳。     

Increased presence of Epstein-Barr virus DNA in ocular fluid samples from HIV negative immunocompromised patients with uveitis

AIMS—To investigate whether routine testing for Epstein-Barr virus (EBV) is necessary in the examination of a patient with uveitis.
METHODS—Intraocular EBV DNA was determined in 183 ocular fluid samples taken from patients with AIDS and uveitis, HIV negative immunocompromised uveitis, acute retinal necrosis, toxoplasma chorioretinitis, intraocular lymphoma, anterior uveitis, and miscellaneous uveitis of unknown cause. In 82 samples from this group of patients paired serum/ocular fluid analysis was performed to detect local antibody production against EBV. Controls (n=46) included ocular fluid samples taken during surgery for diabetic retinopathy, macular pucker, or cataract.
RESULTS—Serum antibody titres to EBV capsid antigen proved to be significantly increased in HIV negative immunocompromised patients with uveitis (p<0.01) compared with controls. Local antibody production revealed only three positive cases out of 82 patients tested, two results were borderline positive and one patient had uveitis caused by VZV. EBV DNA was detected in three out of 46 control ocular fluid samples. In the different uveitis groups EBV DNA was noted, but was not significantly higher than in the controls, except in six out of 11 HIV negative immunocompromised patients (p=0.0008). In four out of these six cases another infectious agent (VZV, HSV, CMV, or Toxoplasma gondii) had previously been identified as the cause of the uveitis.
CONCLUSIONS—When comparing various groups of uveitis patients, EBV DNA was found more often in HIV negative immunocompromised patients with uveitis. Testing for EBV does not have to be included in the routine management of patients with uveitis, since indications for an important role of this virus were not found in the pathogenesis of intraocular inflammation.

Keywords: Epstein-Barr virus; intraocular fluid; polymerase chain reaction; uveitis     

Outcome of abducens nerve paralysis in patients with nasopharyngeal carcinoma

PURPOSE: The objective of this study was to investigate the incidence, management and outcome of patients with nasopharyngeal carcinoma (NPC) who developed Vlth nerve palsy. METHODS: Between December 1993 and December 1999, we investigated retrospectively the charts of 166 patients with NPC. RESULTS: Twenty-eight of 166 patients (16.8%) had cranial nerve involvement at the time of the diagnosis of NPC. Sixteen (57.2%) were identified as having abducens nerve palsy. In 25% abducens nerve palsy was the presenting symptom. Three patients were able to compensate for their diplopia after prism correction or botulinum toxin-A injection, and six (50%) completely recovered from abducens nerve palsy after either radiotherapy or chemotherapy. CONCLUSIONS: Abducens is the most common cranial nerve involved in NPC, radiotherapy and or chemotherapy relieves the paralysis in half the patients. Prism correction or botulinum toxin-A injection are effective non-invasive procedures for patients with significant diplopia.     

Quantitative PCR for the detection of genomic DNA of Epstein-Barr virus in ocular fluids of patients with uveitis

Purpose  To measure the genomic DNA from Epstein-Barr virus (EBV) in ocular fluids and to analyze the clinical relevance of EBV in uveitis. Methods  Intraocular fluids (30 aqueous humor and 30 vitreous fluid samples) were taken from 55 patients with uveitis after informed consent was obtained. Samples were assayed for EBV DNA using qualitative multiplex polymerase chain reaction (PCR) and quantitative real-time PCR. Antibodies to EBV were examined using a complement fixation test. Results  EBV DNA was detected in 17 of 60 samples (28%) and 16 of 55 patients (29%) using multiplex PCR. However, only three of the 17 samples showed significantly high copy numbers of EBV DNA with real-time PCR. EBV DNA was not detected in the serum of all patients. EBV-specific antibodies were positive in the serum of all patients, but not in the vitreous fluid. Vitreous anti-EBV antibodies were positive only in patients displaying genomic DNA of EBV in the vitreous samples. Conclusions  EBV DNA was detected by qualitative PCR in ocular fluids of many uveitis patients, but only a small proportion of patients showed high viral loads on quantitative real-time PCR, indicating that replication of the virus takes place only in a few patients.     

Impairment of optic path due to radiotherapy for nasopharyngeal carcinoma

Purpose: To evaluate the impairment of optic path caused by radiotherapy for nasopharyngeal carcinoma (NPC). Methods: Visual evoked potential was used to evaluate the functional impairment of optic path by an 8 MV linear accelerator or ⁶⁰Co -ray radiotherapy in 63 (23 women and 40 men) patients with nasopharyngeal carcinoma before radiotherapy, at the end of radiotherapy, 6 months and 1, 2 and 3 years after their radiotherapy respectively. Results: In the female group, the latency of VEP elicited by the three types of elements significantly delayed 2 and 3 years after radiotherapy than that before radiotherapy, at the end of radiotherapy, 6 months and 1 year after radiotherapy; there were no significant difference among VEP amplitudes elicited by the three types of elements before and after radiotherapy. In the male group, the latency of VEP elicited by the medium and the small elements significantly delayed 1 year after radiotherapy than that before radiotherapy and at the end of radiotherapy, the VEP latency elicited by the small elements was significantly prolonged 3 years after radiotherapy compared to that just after radiotherapy, while the VEP amplitude elicited by the large elements was significantly attenuated 1 year after radiotherapy compared to those before and just after radiotherapy. Conclusions: In the female group, the significant prolongation of VEP latency happened at the end of radiotherapy and lasted for 3 years, while the VEP amplitude did not change significantly during the 3 years after radiotherapy. It indicates that the sustained impairment by radiation within the female visual nerve system starts at the end of radiotherapy, but the impairment was mild. In the male group, the significant prolongation of VEP latency mostly happened 1 or 2 years after radiotherapy. It was shown that the radiotherapy for nasopharyngeal carcinoma certainly injured the optic path, and there was difference in the impairment between the two sexes.     

Epstein-Barr virus infection and immunologic dysfunction in patients with aqueous tear deficiency

The authors tested their hypothesis that Epstein-Barr virus (EBV) infection is a risk factor for aqueous tear deficiency (ATD) by evaluating 38 ATD patients and 17 controls for serologic evidence of EBV infection. Aqueous tear deficiency was graded clinically as mild or severe. A linear trend toward elevated EBV capsid (P less than 0.05) and early antigen (P less than 0.001) titers was noted from control to severe ATD patients. Rubella and cytomegalovirus antibody titers were poorly correlated with EBV titers, suggesting that the elevated EBV antibodies in ATD patients were not due to nonspecific polyclonal B-cell activation. Epstein-Barr virus antigens were detected in two of six lacrimal gland biopsies from severe ATD patients with Sj?gren's syndrome, but in none of the control glands. Aqueous tear deficiency patients were evaluated for immunologic dysfunction associated with EBV infection. Linear trends of elevated serum IgG (P less than 0.05), autoantibody and immune complex positivity (P less than 0.05), and reduced natural killer cell cytotoxicity (P less than 0.05) were found from controls to severe ATD patients. Furthermore, reduced T-helper lymphocyte counts (P less than 0.06) and an increased percentage of HLA-DR+ CD8 lymphocytes (P less than 0.05) were observed in severe ATD patients compared with the mild and control groups. A multivariate analysis of the data showed a significant correlation between severe ATD and elevated EBV early antigen titers, Sj?gren's syndrome, and an increased percentage of HLA-DR+ CD8 lymphocytes. The authors' findings suggest that EBV infection may be a risk factor for development of ATD in a subset of ATD patients with greater disease severity, Sj?gren's syndrome, and immunologic dysfunction.