人、兔结膜上皮细胞的原代培养

目的探索结膜上皮细胞体外培养的最佳方法,研究其生长特性,为毒理实验及体外结膜上皮移植提供基础。方法分别用3种培养方法：(1)组织块培养法;(2)混合肖化法;(3)组织块消化后贴壁法来培养人、兔的结膜上皮细胞,观察细胞状态、不同部位的长生特性等,并尝试在羊膜、饲细胞层上培养建系。结果3种培养方法中,组织块培养法能获得较纯的结膜上皮细胞但生长缓慢,消化法能获得大量细胞,生长较快,但常常混有其它杂细胞,组织块消化后贴壁法获得细胞量较少。观察到来自兔、睑结膜及穹隆部结膜上皮的细胞生长速度较快,细胞量多,来自球结膜的细胞生长较慢。应用间接免疫荧光染色,人结膜上皮细胞K13染色阳性。结论组织块培养法及混合消化法皆能培养结膜上皮细胞,各有利弊。     

不同方法体外培养兔结膜上皮细胞的比较研究

目的比较不同方法体外培养兔结膜上皮细胞的效率,确定体外培养兔结膜上皮细胞的最佳方法,建立兔结膜上皮细胞系,为进一步研究结膜上皮干细胞的鉴别、定位及其增殖分化提供基础。方法分别采用DispaseⅡ消化法、组织块培养法、胰酶／EDTA混合消化法获得兔结膜上皮细胞后进行体外培养,在不同时间观察细胞的形态,并应用免疫组化的方法进行细胞鉴定。结果组织块培养法细胞生长较慢,胰酶／EDTA混合消化法容易有成纤维细胞等污染,DispaseⅡ化法能够获得大量的较纯净的结膜上皮细胞,细胞生长增殖快,免疫荧光pan-cytoeratin染色阳性．部分细胞MUC5AC染色阳性。结论DispaseⅡ消化法是进行体外结膜上皮细胞培养的最佳方法。     

自体结膜移植治疗翼状胬肉的临床观察

翼状胬肉是眼科常见病 ,治疗最佳方法是手术切除 ,但单纯翼状胬肉切除术术后复发率高达30 %～ 5 0 % [1]。我们自 2 0 0 0 - 0 3～ 2 0 0 1 - 0 2 ,采用自体结膜移植术治疗翼状胬肉 ,取得了满意的疗效 ,现报道如下。1　资料与方法2 6例 ( 2 9眼 )中 ,男 1 1例 ( 1 2眼 ) ,女 1 5例 ( 1 7眼 ) ;右眼 1 6例 ,左眼 1 3例。年龄 32～ 69岁 ,平均5 1岁。胬肉头部侵入角膜内小于 2 mm者 6例 ( 7眼 ) ,侵入角膜内 2～ 4mm者 1 8例 ( 2 0眼 ) ,接近瞳孔缘者 2例 ( 2眼 )。术后随访 6～ 8mo,平均 1 4mo。手术方法 :0 .5 %地卡因表面麻醉后 ,2 %利多卡…     

人眼结膜上皮细胞体外培养治疗翼状胬肉

目的 探讨体外分离培养结膜上皮细胞治疗翼状胬肉的方法.方法 将126例翼状胬肉患者(均为单眼患者)随机分为2组.治疗组58例,术前2周取其穹隆部结膜(1mm×1mm)剪成0．5mm×O．5mm大小后种植于羊膜上体外培养,术中逆行切除胬肉组织,再将培养好的结膜上皮植片移植于结膜创口,缝合固定.对照组68例,单纯切除翼状胬肉,暴露角巩膜缘.两组术后均行妥布霉素眼膏及滴眼液治疗,预防感染.结果 接种在羊膜上的结膜组织,8d左右融合成膜状,细胞为复层上皮细胞.植片3d后开始呈透明或半透明状,1周时可见新生血管长入,植片与周围结膜逐渐融合,2周左右植片与周围结膜完全融合.术后随访1～2年.治疗组复发2例(3．45％),对照组复发23例(33．82％),两组问复发率差异显著,有统计学意义(x2=16．3．P<0．01).所有患者眼球活动自如,未发生睑球粘连、疤痕增生等并发症.结论 体外分离培养结膜上皮细胞治疗翼状胬肉是一种理想的方法.     

结膜角膜缘上皮自体移植治疗翼状胬肉的效果观察

我院1996年6月～1998年12月采用结膜角膜缘上皮自体移植覆盖胬肉切除术后巩膜和角膜缘创面的方法治疗翼状胬肉共38例，取得满意效果。现报告如下。1 临床资料1.1 一般资料：本组38例47眼，其中男14例16眼，女24例31眼。年龄26～78岁，平均49岁。复发性胬肉6例8眼，其余为原发性胬肉，双眼发病共9例。为观察术后反应，均住院治疗。     

结膜下翼状胬肉逆向切除术

目的 评价结膜下翼状胬肉逆向切除术。方法 回顾分忻了54例结膜下翼状胬肉逆向切除术患者。结果 随访发现，治愈53例，复发1例，无并发症出现。结论 结膜下翼状胬肉逆向切除术，显微镜下操作，手术精细，损伤小，复发少，不失为治疗翼状胬肉的一种较好的手术方式。     

翼状胬肉切除后结膜瓣移植术

目的：通过对34例（34眼）角膜缘翼状胬肉切除加结膜瓣移植术后的观察，总结手术方法及疗效。方法：对34例（34眼）行角膜缘翼状胬肉切除加结膜瓣移植术。结果：通过上述方法可将角膜缘翼状胬肉完整切除，无角膜深层损伤，减少复发等优点。结论：翼状胬肉切除加结膜瓣移植术方法操作相对简单，患者容易接受，疗效满意，是一种有效的治疗方法。     

结膜瓣移植治疗翼状胬肉

目的观察结膜移植治疗翼状胬肉的临床疗效。方法采用翼状胬肉切除联合结膜移植，部分复发性翼状胬肉采用翼状胬肉切除及自体角膜缘上皮联合球结膜移植治疗翼状胬肉。观察术后角膜愈合、角膜缘新生血管及球结膜是否有翼状胬肉样组织。结果术后随访36～72个月，有5眼复发，复发率为6．58％。结论自体球结膜移植治疗翼状胬肉能有效地减少翼状胬肉的术后复发率，是治疗翼状胬肉并防止复发的有效方法。     

结膜延伸移植术治疗71例翼状胬肉的体会

采用结膜延伸移植术治疗翼状胬肉71例89只眼,术后半年随访,3例复发,其余均治愈,治愈率达96.6％。作者认为,复发原因多为手术时对結膜下胬肉组织未作剥离或切除;对巩膜及角膜的胬肉组织切剥不彻底;胬肉分离或切除后,对结膜的创缘未作缝合固定。     

结膜上皮转位与角膜缘上皮移植治疗复发性翼状胬肉的疗效对比

翼状胬肉手术切除术后复发率较高。复发性翼状胬肉瘢痕组织增多增厚，甚至出现眼球运动障碍，形成复视，我们自2000年开始对复发性翼状胬肉患者分别采用结膜上皮转位及角膜缘上皮移植治疗，现将结果报告如下。     

Inhibitory Effect of Curcumin on Proliferation of Human Pterygium Fibroblasts

In order to investigate the effect of curcumin on proliferation and apoptosis of human pterygium fibroblasts (HPF) in culture and search for a new method to prevent the recurrence after pterygium surgery, HPF was incubated with 0-160 μmol/L curcumin for 24-96 h. The MTT method was used to assay the biologic activities of curcumin at different time points and different doses. The expression of proliferating cell nuclear antigen (PCNA) in each group was detected by immunohistochemistry. The cell cycle distribution was detected by flow cytometry (FCM). Admini- stration of 20-80 μmol/L curcumin for 24-72 h could significantly inhibit HPF proliferation in a dose- and time-dependent manner (P<0.05). After treatment with curcumin at different concentrations of 20, 40, 80 and 160 μmol/L for 24 h, FCM revealed there was a significant sub-G1 peak at each concentration. The number of HPF in G0/G1 phase was increased, while in S phase, it was decreased (P<0.05). At the concentration of 20-80 μmol/L, curcumin, in a dose-dependent manner (P<0.05), could inhibit the expression of PCNA in HPF. It was suggesterd that curcumin could significantly in- hibit the proliferation of HPF, make HPF arrest in G0/G1 phase and induce the apoptosis of HPF in a dose- and time-dependent manner.     

老年翼状胬肉患者翼状胬肉切除联合结膜移植术后绷带式角膜接触镜的应用及效果评价

目的：探讨老年人翼状胬肉切除联合结膜移植术后应用绷带式角膜接触镜（BCL）的临床疗效,并阐明该疗法的优势。方法：回顾性分析本院70例70眼老年翼状胬肉患者的临床资料,按术后处理方法分为2组,观察组（BCL组）35例35眼,行翼状胬肉切除联合结膜移植术,术毕术眼立即配戴BCL;对照组35例35眼,术毕常规纱块敷料包眼。观察患者术后1、3和7和10 d视物模糊、畏光、异物感、流泪和眼痛的症状评分、角膜荧光素染色（FSC）评分和角膜上皮缺损面积。结果：术后第1、3和7天,观察组患者视物模糊、畏光、异物感、流泪和眼痛症状评分均低于对照组（P<0.05）。术后第3和7天,观察组患者FSC评分和角膜上皮缺损面积均低于对照组（P<0.05）。术后10 d 2组患者不适症状、FSC评分和角膜上皮缺损面积比较差异无统计学意义（P>0.05）。随访期间,所有患者均未发生角膜感染和植片脱落等并发症。结论：BCL能明显缓解老年人翼状胬肉切除术后的不适感,促进角膜上皮愈合。     

HDAC1 expression and effect of curcumin on proliferation of Raji cells

Summary Histone deacetylase (HDAC₁) has a high expression in many cancer cells and curcumin can inhibit the growth of cancer cells. This paper was designed to investigate the expression of HDAC₁ of Raji cells and the effect of curcumin on their proliferation and apoptosis. Raji cells were treated with 3. 125–50 μmol/L curcumin for 8–48 h and the growth inhibition rates of Raji cells were measured by MTT. The expression of HDAC₁ on Raji cells were examined by mRNA, Western blot at 24 h various concertrations (1.6–50 μmol/L). Curcumin could selectively inhibit the proliferation of Raji cells in a dose and time dependent manner, with the inhibition rate being 52.47%–82.18% (P<0.01). The up-regulation of HDAC₁ expression was observed within 24 h after the treatment with curcumin as shown by RT-PCR and Western blot. With the increase of concentration, the expression was down-regulated in a dose dependent manner. It is concluded that the expression of HDAC₁ plays an important role in the proliferation and apoptosis of Raji cells and curcumin can inhibit the growth of Raji cells at various concentrations and promote the apoptosis of Raji cells. WU Qing, female, born in 1970, M. D., Ph. D. This project was supported by a grant from the National Natural Sciences Foundation of China (No. 30271672).     

姜黄素联合人类细胞因子诱导的杀伤细胞对卵巢癌细胞增殖的抑制作用及其机制

目的:观察姜黄素(Cur)联合人类细胞因子诱导的杀伤(CIK)细胞在体外对人卵巢癌浆液性囊腺癌细胞SKOV-3的增殖抑制作用,探讨其协同抗肿瘤作用及其可能机制.方法:诱导脐血CIK细胞,将SKOV-3细胞随机分为Cur组、CIK细胞组和Cur联合CIK细胞组,MTT法测定各组细胞的增殖抑制率,RT-PCR检测各组细胞F...     

姜黄素在体外对原代白血病细胞的作用

目的 :研究姜黄素在体外对原代白血病细胞的抑制作用。方法 :MTT法 ,台盼蓝拒染法。结果 :姜黄素对临床白血病患者原代白血病细胞初发组抑制率显著大于复发组 (P <0 .0 5 )。结论 :姜黄素对原代白血病细胞具有一定的抑制作用     

姜黄素对翼状胬肉成纤维细胞抑制作用的研究

目的研究姜黄素对体外培养人翼状胬肉成纤维细胞（human pterygium fibroblasts HPF）增殖的影响,探索翼状胬肉防治新方法。方法用0~80μg/mL姜黄素作用体外培养的人翼状胬肉成纤维细胞,观察24~96h后不同浓度姜黄素对人翼状胬肉成纤维细胞的影响。CCK-8法检测细胞生长活性,流式细胞仪检测细胞周期时相变化。结果在0~80μg/mL浓度作用24~96h范围内,姜黄素可以抑制HPF细胞的增长,且呈剂量和时间依赖性。0、10、20、40、80μg/mL姜黄素作用24h后,姜黄素作用组均出现G0-G1期细胞百分比上升,S期细胞百分比下降,且姜黄素对成纤维细胞的抑制作用呈剂量依赖性。结论姜黄素可抑制体外培养人翼状胬肉成纤维细胞增殖。     

兔眼结膜上皮细胞和成纤维细胞的培养及冻存

目的 探讨体外培养兔眼结膜上皮细胞和成纤维细胞的最佳方法,为进一步研究结膜的生物学特性和结膜重建提供基础。 方法 分别用组织块法培养结膜上皮细胞和混合消化法培养结膜基质细胞,用倒置显微镜观察其生长方式和形态特征;收集第2代细胞,-80℃冻存。结果 兔眼结膜上皮细胞和上皮下成纤维细胞可以在体外成功培养,结膜上皮细胞呈圆形或多角形,成纤维细胞呈长梭形;冻存细胞1个月后复苏成功率达80%。结论 组织块法和混合消化法培养可成功获得结膜上皮细胞和上皮下成纤维细胞。     

丝裂霉素C联合自体游离结膜瓣移植治疗翼状胬肉的临床观察

目的探讨翼状胬肉切除并自体游离结膜瓣移植联合丝裂霉素C(MMC)治疗翼状胬肉后出现并发症的原因、治疗及预防。方法对139例翼状胬肉患者(153眼)行翼状胬肉切除联合自体结膜瓣移植,术中MMC(0.2 mg/mL)棉片于巩膜面湿敷2 m in,随访1年,对术后并发症进行分析。结果胬肉复发6眼(3.92%);结膜创口延迟愈合11眼(7.19%)及浅层点状角膜炎9眼(5.88%),未予特殊治疗;结膜瓣溶解7眼(4.58%)、巩膜无血管化2眼(1.31%)及继发青光眼1眼(0.65%),局部药物治疗有效;结膜肉芽肿2眼(1.31%)及角巩膜溶解患者2眼(1.31%),联合手术治疗。结论联合丝裂霉素C治疗翼状胬肉可降低胬肉复发,但同时可能造成角巩膜溶解等并发症。应术前排除免疫性疾病史,术中注意敷贴放置位置,术后注重随访,以减少并发症发生。     

Efficacy of treatment for pterygium by autologous conjunctival transplantation and mitomycin C

Background  The recurrence of pterygium after surgery is high. A variety of adjunctive treatment approaches have been developed to improve the clinical efficacy and many related articles have been published. This study aimed to determine the risk for postoperative pterygium recurrence comparing autologous conjunctival transplantation (ACT) versus mitomycin C (MMC).

Methods  Relevant literature published until December 2010 in Medline, Embase, Cochrane, Cochrane library, Science Citation Index, and Google Scholar were searched. Qualified random clinical trial (RCT) studies on the comparison of recurrence rate of pterygium after ACT and MMC treatment were included in this study.

Results  Eight RCTs with 663 eyes entered the final analysis. The recurrent rate of pterygium was 8.7% (30/343) for ACT and 18.75% (60/320) for MMC. Using fixed-effect meta analysis, we found that the recurrence was significantly lower after ACT than MMC treatment (odds ratio (OR)=0.40, 95% confidence index (CI), 0.25–0.63, P <0.0001). In sensitivity analyses, we employed random-effects model and excluded studies of low quality or studies in which MMC was administrated after the operation. All the sensitivity analyses confirmed that ACT led to lower recurrence rates than MMC. Sub-group analysis revealed that the recurrence rate was 20.2% (20/99) and 27.65% (26/94) for conjunctival autograft (CA) and MMC respectively, and no significant difference in the recurrence rate was detected (OR=0.65, 95% CI 0.33–1.28, P=0.22). However, we found that conjunctival limbal autograft (CLA) had lower recurrence rate than MMC (OR=0.26, 95% CI 0.14–0.48, P=0.0001).

Conclusion  CLA has better therapy efficacy against the recurrence of pterygium than MMC.