Addressing the MSICS learning curve: identification of instrument-holding techniques used by experienced surgeons

AIM: To detect the quantitative expression levels of the pro-inflammatory interleukin-8 (IL8), antimicrobial peptides Human beta defense-2 (HBD2), and Human beta defense-3 (HBD3) genes in bacterial conjunctivitis. METHODS: The human conjunctival epithelial cells were obtained using the impression cytology technique from healthy controls and patients. The genes expression levels were determined utilizing a quantitative real-time polymerase chain reaction (RT-PCR). The contribution of causative agent type, the number of isolates and severity of clinical features, in the increase of genes expression was also determined. RESULTS: The RT-PCR showed that IL8, HBD2, and HBD3 expression increased in bacterial conjunctivitis as compared to healthy control (P<0.001). In gram-negative bacterial conjunctivitis, HBD2 was highly up-regulated (P<0.001) compared to other types of bacterial conjunctivitis. In mixed bacterial conjunctivitis, a direct correlation between HBD2 up-regulation and HBD3 up-regulation was observed (P<0.05). The severity of clinical features was related to the up-regulation of IL8 and HBD2 (P<0.05). CONCLUSION: IL8, HBD2, and HBD3 are immune-effectors in infectious conjunctivitis. HBD2 is active during different bacterial conjunctivitis but is more released with gram-negative bacteria compared to gram-positive bacteria. HBD3 is an obvious defender in different bacterial conjunctivitis.     

Is it possible to detect the presence of adenovirus in conjunctiva before the onset of conjunctivitis?

Purpose: It is widely accepted that patients with adenoviral conjunctivitis are contagious approximately as long as 14 days after the onset. However, there is controversy about whether patients with adenoviral conjunctivitis are contagious during the incubation period. We investigated the potential of the eye as a source for spreading infection during the incubation period by analysing the presence of adenovirus (Ad) in contralateral eyes before onset in patients with bilateral adenoviral conjunctivitis using the immunochromatography (IC) test and polymerase chain reaction (PCR). Methods: Patients with adenoviral conjunctivitis were directed to visit an outpatient clinic at approximately 3‐day intervals until onset in the contralateral eye or 2 weeks after onset in the first eye. Conjunctival scrapings obtained from the first infected eye and the contralateral eye before and after onset were tested to detect Ad antigen using the IC test and PCR. Results: Out of the 32 cases, 23 (72%) developed conjunctivitis in the contralateral eye. Among these bilateral cases, 18 (78%) were found positive for Ad by PCR after onset in the contralateral eye. However, no specimen obtained before onset showed a positive result for Ad in both tests. Conclusion: Although there is a possibility that eyes are contagious during the incubation period of adenoviral conjunctivitis in some situations, these results suggest that the possibility of the eye acting as a source for spreading infection during the incubation period is very small.     

Polymerase chain reaction in the diagnosis of bacterial endophthalmitis

BACKGROUND—Microbiological investigations of vitreous fluid (VF) and aqueous humour (AH) specimens have often failed to detect the infecting agent in infectious endophthalmitis, resulting in a clinical dilemma regarding therapy. In this study, the polymerase chain reaction (PCR) was evaluated in the diagnosis of bacterial and Propionibacterium acnes endophthalmitis.
METHODS—58 intraocular specimens (30 VF and 28 AH) from 55 cases of endophthalmitis and 20 specimens (14 VF and 6 AH) as controls from non-infective disorders were processed for microbiological investigations. Nested PCR directed at the 16S rDNA using universal primers for eubacterial genome was done. PCR for P acnes was performed on specimens microbiologically negative by conventional techniques but eubacterial genome positive.
RESULTS—Of the 20 controls from non-infective cases, one (5%) was positive using eubacterial primers and none with P acnes primers. PCR for eubacterial genome showed 100% correlation with 20 (34.5%) bacteriologically positive specimens. Eubacterial genome, was detected in 17 (44.7%) of 38 bacteriologically negative specimens and nine (52.9%) out of the 17 were positive for P acnes genome. Among the 21 eubacterial PCR negative specimens, seven were fungus positive. By inclusion of PCR, microbiologically positive specimens increased from 46.5% to 75.8%. PCR on AH was as sensitive as that on VF for the detection of both eubacterial and the P acnes genome.
CONCLUSION—PCR performed on AH and VF is a reliable tool for the diagnosis of bacterial and P acnes endophthalmitis particularly in smear and culture negative specimens.

Keywords: polymerase chain reaction; bacterial endophthalmitis; infectious endophthalmitis     

复发性单纯疱疹性角膜炎患者血清IL-2及SIL-2R水平的研究

目的研究复发性单纯疱疹性角膜炎患者血清白细胞介素-2（IL－2）及其可溶性白细胞介素-2受体（SIL－2R）水平。方法本文测定了30例复发性单纯疱疹性角膜炎患者（实验组）和25例健康人（对照组）血清白细胞介素－2（IL－2）及其可溶性白细胞介素－2受体（SIL－2R）的水平。结果对照组IL－2的水平为5．41±1．45mg·L－1,实验组IL－2R水平为2．80±1.47mg·L-1,2组比较t＝2．90,P＜0．01,说明复发性单纯疱疹性角膜炎患者血清IL－2水平较对照组明显降低。对照组SIL－2R水平为90．86±17．90mg·L－1,实验组SIL－2R水平为171·46±19.30mg·L－1,2组比较t＝5．17,P＜0．01。复发性单纯疱疹性角膜炎患者血清SlL－2R水平较对照组显著升高。结论复发性单纯疱疹性角膜炎患者体内免疫功能紊乱,血清IL－2活性降低、SlL－2R水平升高可能是其原因之一,这也为免疫治疗提供了一定依据。     

息肉状脉络膜血管病变患者房水中IL-8及其受体的表达

目的：探究息肉状脉络膜血管病变(PCV)患者房水中IL-8及其受体1、2(CXCR1、CXCR2)的水平,并探讨其临床意义。

方法：选取2016-04/2018-03本院收治的PCV患者67例作为PCV组,另选取白内障患者50例作为对照组。采用实时荧光定量PCR法检测两组患者房水中IL-8、CXCR1、CXCR2 mRNA表达量,采用国际标准视力表测量所有受试者BCVA,OCT检测CMT,创建受试者工作特征曲线(ROC)分析IL-8、CXCR1、CXCR2对PCV的诊断价值; 采用Logistic多因素回归分析评估PCV发生的危险因素。

结果：与对照组相比,PCV组患者房水中IL-8、CXCR1、CXCR2 mRNA表达升高,BCVA降低,CMT升高(均P<0.05)。PCV组患者房水中IL-8、CXCR1、CXCR2水平与BCVA(LogMAR)呈正相关(r=0.438、0.346、0.385,均P<0.05),与CMT呈正相关(r=0.378、0.606、0.357,均P<0.05)。IL-8、CXCR1、CXCR2、BCVA、CMT是PCV发生的危险因素。IL-8、CXCR1、CXCR2诊断PCV的ROC曲线下面积(AUC)分别为0.882、0.860、0.812。

结论：PCV患者房水中IL-8、CXCR1、CXCR2表达均升高,其不仅与患者BCVA和CMT相关,且与PCV的发生有关,可能成为PCV的潜在预测指标。     

兔眼IOL术后房水及泪液中白介素-8、钙离子的动态变化

目的研究晶状体超声乳化人工晶状体植入术（phaco＋IOL）及晶状体囊外摘出人工晶状体植入术（ECCE＋IOL）术后房水及泪液中自介素-8（IL-8）、钙离子（Ca^2＋）动态变化及其临床意义。方法30只青紫蓝家兔，随机分为3组（phaco＋IOL组，ECCE＋IOL组，正常对照组），每组10只。均为右眼。术后抽取房水和收集泪液。用比色法测定Ca^2＋，用放射免疫法测定IL-8。结果术后房水中IL-8活性第3天达高峰（P〈0．01），术后第3天Ca^2＋含量最低（P〈0．01）。术后各组泪液中IL-8术后第7天达高峰（P〈0．01），手术各组泪液中Ca^2＋在术后第3天达最低（P〈0．01）。房水与泪液中Ca^2＋、IL-8水平呈直线正相关（P〈0．05）．结论IL-8、Ca^2＋参与术后眼内炎症反应，检测泪液中IL-8和Ca^2＋的含量，能推断房水中IL-8、Ca^2＋含量，可作临床病情监测和指导用药的指标。     

Comparison of uveitis induced by interleukin-8 (IL-8) and endotoxin in rabbits

IL-8 is a potent chemoattractant which has been postulated to play a role in the cytokine cascade associated with uveitis. The authors studied the effect of intravitreal IL-8 on the induction of uveitis in the rabbit. IL-8 at varying concentrations (1 ng, 10 ng or 100 ng) or endotoxin (100 ng) was injected intravitreally within the rabbit eye. At 6, 24 and 48 hours following injection the induction of uveitis was evaluated by clinical scoring, anterior chamber (AC) leukocyte count, AC protein concentration and histopathology in 15 rabbits. Only the 100 ng concentration of IL-8 induced uveitis at 6 and 24 hours by clinical scoring and AC leukocyte count; the AC protein concentration remained normal. In contrast, endotoxin caused a severe uveitis with a significant increase in all the parameters evaluated. The authors conclude that intravitreal IL-8, in the concentrations studied, induces a limited uveitis which is detectable at six hours and resolves within 48 hours. It is characterized by leukocyte infiltration without an increased AC protein concentration. Thus, IL-8 may play a role in the cytokine cascade involved in the induction of uveitis     

IL-23和IL-17在糖尿病性视网膜病变患者房水中的表达

目的：观察年龄相关性白内障合并糖尿病性视网膜病变(DR)患者房水中白细胞介素-23(IL-23)和白细胞介素-17(IL-17)的表达水平及二者的相关性。

方法：收集2016-06/2019-06我院眼科收治的因年龄相关性白内障需行白内障手术的患者70例70眼,其中单纯年龄相关性白内障患者20例(Ⅰ组),年龄相关性白内障合并单纯糖尿病者18例(Ⅱ组),合并非增殖性糖尿病性视网膜病变者17例(Ⅲ组),合并增殖性糖尿病性视网膜病变者15例(Ⅳ组)。采用ELISA法检测各组患者房水中IL-23、IL-17表达水平。

结果：四组患者房水中IL-23的表达水平分别为22.18±5.48、37.63±4.52、45.06±4.64、68.89±6.11pg/mL,IL-17的表达水平分别为4.69±2.03、6.83±1.02、9.52±1.30、10.89±1.26pg/mL,房水中IL-23和IL-17的表达水平随着DR的严重程度呈增高趋势,且糖尿病患者房水中IL-17的表达水平与IL-23呈正相关。

结论：房水中IL-23和IL-17的高表达可能协同促进DR的发生发展,IL-23/IL-17通路作为致病因素,可能通过加重视网膜炎症反应参与DR的病理损伤过程,这为DR的早期诊断和治疗提供了新的靶点。     

Study of the prevalence and association of ocular chlamydial conjunctivitis in women with genital infection by Chlamydia trachomatis, Mycoplasma genitalium and Candida albicans attending outpatient clinic

AIM: To determine the association between chlamydial conjunctivitis and genital infection by Chlamydia trachomatis, Mycoplasma genitalium and Candida albicans, in addition to the possible relationship between cultured bacterial pathogens and oculogenital chlamydial infection. METHODS: This study was performed on 100 (50 symptomatic and 50 asymptomatic) women attending the Gynecological and Obstetric outpatient clinic of Alzahra hospital, Alazhar University. Simultaneously a conjunctival swab was taken from these patients. Polymerase chain reaction (PCR) was done on DNA extracted from both vaginal and conjunctival swab samples. Culture for both vaginal and conjunctival swabs was also done. RESULTS: Candida albicans was the predominant organism isolated by culture in 20% and 40% of conjunctival and vaginal swabs respectively. By the PCR method, ocular Chlamydia trachomatis was present in 60% of symptomatic women, while genital Chlamydia trachomatis infection was present in 30% of symptomatic women. The results of this method also indicated that 25/50 (50%) vaginal swabs were positive with PCR for Candida albicans versus 15/50 (30%) were PCR positive in conjunctival swab. Mycoplasma genitalium was present in only 10% of vaginal swabs. Concomitant oculogenital PCR positive results for Chlamydia trachomatis and Candida albicans were 30% and 28% respectively. CONCLUSION: Ocular Chlamydia trachomatis was associated with genital Chlamydia trachomatis in a high percentage of women followed by Candida albicans. Cultured bacterial organisms do not play a role in enhancement of Chlamydia trachomatis infection.     

Polymerase Chain Reaction and Goldmann-Witmer Coefficient Testing in the Diagnosis of Infectious Uveitis in HIV-Positive and HIV-Negative Patients in South Africa

Purpose: To use polymerase chain reaction (PCR) and Goldmann-Witmer coefficient (GWC) calculation to diagnose infectious uveitis.

Methods: Prospective cross-sectional study.

Results: Twenty-seven of 106 patients had positive PCR and/or GWC results on aqueous humor (AH) sampling and 15 of 27 (55.6%) were HIV-positive. Patients with non-anterior uveitis (NAU) were more likely to be HIV+ (p = 0.005). More than 1 possible pathogen was identified in 9 of 27 patients of whom 7 were HIV+. The final clinical diagnosis was discordant with AH findings in 9 of 27 cases. A positive EBV PCR result was associated with a discordant diagnosis (p = 0.001). All cases of herpetic anterior uveitis (42.9% HIV+) tested PCR-/GWC+ while all cases of herpetic NAU tested PCR+/GWC- (83.3% HIV+). All rubella virus cases were PCR+/GWC+.

Conclusion: PCR is useful to diagnose herpetic NAU in HIV+ patients while GWC is useful to diagnose herpetic anterior uveitis.     

结合磁珠hRPE细胞受附加磁场作用后MCP-1和IL-8的表达

目的:观察磁场对结合磁珠培养的人视网膜色素上皮细胞(hRPE)单核细胞趋化蛋白-1(MCP-1)和白细胞介素-8(IL-8)的表达,以模拟在原发性视网膜脱离形成早期,hRPE受到机械牵拉力后的一些细胞因子分泌的改变。方法:将培养的hRPE与黏附有胶原包被磁珠的培养hRPE分别置于一个附加磁场下,分别暴露一定时间后,用反转录—聚合酶链反应(RT-PCR)方法测定hRPE,黏附包被胶原磁珠的hRPE,及其在细胞松弛素D处理后,各时间点MCP-1和IL-8的表达。结果:只受磁场作用,未结合磁珠的hRPE表达的MCP-1及IL-8mRNA水平较低,不随暴露于磁场时间的延长而增加;在磁场作用下,结合磁珠的hRPE表达MCP-1及IL-8的有两个峰值,一是在暴露后0.5h内开始升高,分别达到3.30±0.12和1.88±0.08(P=0.009),另一是在4h左右时再次升高。细胞松弛素D预处理后,两种因子的表达均下降,分别为2.36±0.27和1.64±0.08。结论:hRPE细胞结合磁珠模拟的机械牵拉模型,可诱导细胞表达MCP-1和IL-8增加。这种作用可能是细胞骨架被部分破坏,短时间地抑制了细胞表达。我们的结果提示这两种炎性细胞趋化因子可能参与了视网膜脱离起始阶段的炎症反应过程。     

LED光源对人视网膜色素上皮细胞分泌单核细胞趋化因子-1和白细胞介素-8的影响

目的　观察ＬＥＤ光源对人视网膜色素上皮（ｒｅｔｉｎａｌｐｉｇｍｅｎｔｅｐｉｔｈｅｌｉａｌｃｅｌｌｓ,ＲＰＥ）细胞增生及分泌单核细胞趋化因子-１（ｍｏｎｏｃｙｔｅｃｈｅｍｏｔａｃｔｉｃｐｒｏｔｅｉｎ-１,ＭＣＰ-１）和白细胞介素-８（ｉｎｔｅｒｌｅｕｋｉｎ-８,ＩＬ-８）的影响。方法　传代培养的ＡＲＰＥ细胞分别置于５００ｌｕｘ、１０００ｌｕｘ的ＬＥＤ白光、蓝光、绿光中分别照射６ｈ、１２ｈ、２４ｈ,并分别设置对照组避光培养。采用ＭＴＴ法检测ＲＰＥ细胞的增生值（Ａ４６０）,分别使用Ｒｅａｌ-ｔｉｍｅＰＣＲ和ＥＬＩＳＡ法检测各组ＲＰＥ细胞ＭＣＰ-１和ＩＬ-８的表达水平。结果　ＭＴＴ法检测细胞增生值显示:蓝光、白光各组比较差异均有统计学意义（均为Ｐ＜０．０５）,白光５００ｌｕｘ２４ｈ和１０００ｌｕｘ１２ｈ、２４ｈ,蓝光各照度６ｈ、１２ｈ、２４ｈ,ＲＰＥ细胞增生值均较对照组低,差异均有统计学意义（均为Ｐ＜０．０５）,细胞增生值随着白光和蓝光光照强度及光照时间的增加逐渐下降。ＲＴ-ＰＣＲ结果显示,各组ＲＰＥ细胞中ＭＣＰ-１和ＩＬ-８的ｍＲＮＡ比较差异均有统计学意义（均为Ｐ＜０．０５）。其中白光５００ｌｕｘ２４ｈ和１０００ｌｕｘ２４ｈ,蓝光５００ｌｕｘ１２ｈ、２４ｈ和１０００ｌｕｘ１２ｈ、２４ｈ,绿光５００ｌｕｘ２４ｈ和１０００ｌｕｘ２４ｈ,各组ＲＰＥ细胞中ＭＣＰ-１和ＩＬ-８的ｍＲＮＡ表达水平较对照组高,差异均有统计学意义（均为Ｐ＜０．０５）。ＥＬＩＳＡ结果显示,白光５００ｌｕｘ２４ｈ和１０００ｌｕｘ１２ｈ、２４ｈ,蓝光５００ｌｕｘ１２ｈ、２４ｈ和１０００ｌｕｘ１２ｈ、２４ｈ,绿光５００ｌｕｘ２４ｈ、１０００ｌｕｘ２４ｈ,与对照组相比, ＲＰＥ细胞ＭＣＰ-１、ＩＬ-８分泌量增加,差异均有统计学意义（均为Ｐ＜０．０５）;蓝光在５００ｌｕｘ２４ｈ和１０００ｌｕｘ１２ｈ、２４ｈ,ＭＣＰ-１和ＩＬ-８分泌量较白光、绿光增加,差异均有统计学意义（均为Ｐ＜０．０５）。结论　ＬＥＤ白光、蓝光和绿光能以照度和时间依赖性影响ＡＲＰＥ细胞增生及分泌ＭＣＰ-１和ＩＬ-８,以蓝光更为显著。     

硫酸镍与芪归解毒汤对人晶状体上皮细胞中HSP27表达的影响

目的:探讨硫酸镍损伤后人晶状体上皮细胞(lens epithelial cell,LECs)中热休克蛋白27(heat shock protein27,HSP27)表达情况,并给予芪归解毒汤干预,观察晶状体上皮细胞HSP27表达的变化。方法:体外培养正常人晶状体上皮细胞分为正常组,硫酸镍组和中药组,分别作用24h后,逆转录聚合酶链反应(RT-PCR)检测晶状体上皮细胞HSP27 mRNA的表达水平。结果:硫酸镍损伤可造成晶状体上皮细胞HSP27 mRNA表达水平上调(P<0.05),预先的中药干预能使HSP27 mRNA表达水平上调(P<0.05)。结论:硫酸镍损伤后晶状体上皮细胞HSP27mRNA表达水平上调,给予预先的中药干预可能通过提高HSP27的表达水平从而保护晶状体上皮细胞。     

白介素-18及信号转导和转录激活因子5在 糖尿病大鼠视网膜的表达

目的观察白介素-18（IL-18）及信号转导和转录激活因子5（STAT5）在4 ～24周糖尿病大鼠视网膜的表达，初步探讨其在糖尿病视网膜病变（DR）中可能的分子机制。方法用限制性片段差异显示聚合酶链反应（RFDD-PCR）技术,建立正常大鼠和糖尿病8周大鼠视网膜基因表达谱。以生物信息学分析两者差异，筛选出候补基因IL-18及STAT5。以半定量反转录聚合酶链反应（RT-PCR）观察IL-18及STAT5在4、8、24周糖尿病大鼠视网膜的表达。结果RFDD-PCR结果显示，正常组IL-18表达明显强于糖尿病组；正常组STAT5无表达，糖尿病组较强表达。RT-PCR结果显示，与正常相比，糖尿病4周时，视网膜高表达IL-18，8周IL-18表达降低，24周表达最低。 STAT5在正常及糖尿病4周视网膜未见表达； 8周开始表达，24周时最强。结论IL-18表达变化及STAT5的活化与DR发生有关。IL-18的表达不依赖于STAT5的活化。(中华眼底病杂志,2005,21:258-260)     

FOXO3和IL-2在干眼患者结膜上皮细胞及泪液中表达及意义

目的:探究叉头框蛋白O3(FOXO3)、白细胞介素-2(IL-2)在干眼(DE)患者结膜上皮细胞及泪液中的表达及意义。方法:前瞻性研究。选择2019-03/2021-03收治的DE患者106例,另选同期85位健康体检者为对照。利用实时荧光定量PCR(qRT-PCR)法检测结膜上皮细胞及泪液中FOXO3水平;酶联免疫吸附(ELISA)法检测样本中IL-2水平;分析DE患者治疗前后的泪膜破裂时间(BUT)、泪液分泌试验(SⅠt)、角膜荧光素染色(CFS)等眼表临床指标变化;Pearson相关性分析DE患者结膜上皮细胞、泪液中FOXO3和IL-2水平的相关性以及二者与临床指标的关系。结果:与对照组相比,DE组患者结膜上皮细胞及泪液中FOXO3水平均明显降低,IL-2水平均明显升高(均P<0.01)。与治疗前相比,DE患者治疗后结膜上皮细胞及泪液中FOXO3水平明显上调,IL-2水平明显下调(均P<0.01)。Pearson相关性分析结果显示,结膜上皮细胞、泪液中FOXO3和IL-2水平均呈显著负相关(r=-0.531、-0.469,均P<0.01)。DE患者治疗后BUT、S...     

22例中国Coats病患儿NDP基因检测分析

目的 针对国外有报道认为NDP基因SNP与Coats病发病可能相关,观察22例中国Coats病患儿NDP基因外显子突变情况,探讨其在Coats病发病机制中的作用。设计 基因研究。研究对象 22例Coats病患儿及6例正常对照。方法 将全部22例患儿和6例正常对照样品的基因组DNA经聚合酶链反应扩增的产物进行测序,分析检测NDP基因全外显子突变情况。主要指标 基因序列。结果 在NDP基因的3个外显子中,未发现有意义的SNP位点,仅发现c.943T＞C（9号、18号）及c.943T＞Y（27号、28号,Y代表C/T杂合子）,但上述突变均位于蛋白质编码区外。结论 22例患儿均未发现有意义的NDP基因SNP,中国Coats病患儿的发病可能与NDP基因突变无关。（眼科, 2016, 25： 127-129）     

大鼠钝挫性眼外伤动物模型中晶状体上皮细胞热休克蛋白70和27的表达及其热耐受的调节作用

目的探讨大鼠眼钝挫伤后晶状体上皮细胞HSP70和HSP27的表达,并给予预先的热刺激(热休克),观察热休克对晶状体上皮细胞HSP70和HSP27表达的调节。方法48只Sprague-Dawley(SD)大鼠,应用随机数字表法随机分成扑打组和水浴组,每组24只鼠,右眼为实验眼。扑打组:20g铁球20cm高度拍打眼球100次。水浴组:给大鼠体温提高至40.5~41.5℃8min诱导出热耐受,2~3h后再给予20g铁球20cm高度拍打眼球100次。逆转录聚合酶链反应检测晶状体上皮细胞HSP70和HSP27基因的表达丰度。结果钝挫性眼外伤可造成晶状体上皮细胞HSP70基因表达的增强,扑打眼球后1h HSP70表达升高至4.59±0.12,3h后达到高峰7.72±0.27,24h后降至正常1.32±0.14。预先的热刺激导致HSP70基因表达增高至1.83±0.30。两组HSP27基因表达丰度均无明显改变。结论钝挫性眼外伤中晶状体上皮细胞HSP70表达的增强提示HSP70可能在钝挫性外伤过程中对晶状体变性蛋白起保护作用,给予预先的热刺激可能通过提高HSP70的表达保护晶状体。(中华眼科杂志,2006,42:241-245)