祛疣洗剂治疗尖锐湿疣前后表皮细胞的增殖及凋亡的变化

目的:探讨祛疣洗剂治疗尖锐湿疣(CA)的作用机制.方法:应用免疫组化SP法检测祛疣洗剂治疗前后基底细胞和棘细胞的Ki-67及表皮Bax、Bcl-2的表达水平,同时采用TUNEL法检测治疗前后表皮细胞的凋亡情况.结果:祛疣洗剂治疗后基底细胞和棘细胞增殖指数明显低于治疗前(P<0.05),而Bax、Bcl-2在祛疣洗剂治疗前后表皮的表达无明显差异(P>0.05),细胞凋亡指数在治疗后略高于治疗前,但无显著性差异(P>0.05).结论:祛疣洗剂治疗CA的机制可能是通过抑制表皮的基底细胞和棘细胞的增殖而发挥作用的.     

尖锐湿疣细胞凋亡与caspase-3、bcl-2的表达

目的探讨尖锐湿疣（CA）细胞凋亡与caspase-3、bcl-2的表达及其相互关系。方法对33例CA,5例巨大CA和8例正常上皮用TUNEL法原位检测角质形成细胞（KC）的凋亡,用免疫组化法检测caspase-3、bc1-2的表达。结果在CA和巨大CA两组之间。凋亡指数（AI）、caspase-3及bcl-2的阳性表达率均无统计学差异（P〉0．05）。CA组的AI与caspase-3的表达水平呈极显著正相关（r=0．48979,P〉0．0001）,与bcl-2的表达水平呈极显著负相关（r=-0．51470,P〉0．0001）,caspase-3与bcl-2的表达水平呈极显著负相关（r=-0．70165,P〉0．0001）。,结论CA的KC中存在着明显的细胞凋亡现象。caspase-3和bcl-2可能参与了CA细胞凋亡的调节机制．     

PCNA、P21和COX-2在尖锐湿疣中的表达及其意义

目的：研究尖锐湿疣组织中增殖细胞核抗原（PCNA）、P21及环氧化酶-2（COX-2）蛋白表达及其意义。方法：应用免疫组化SP法对30例尖锐湿疣组织及10例正常组织中PCNA、P21及COX-2蛋白进行检测。结果：30例尖锐湿疣组织中PCNA、P21阳性表达较正常对照组有不同程度的增加，有统计学意义；COX-2蛋白在尖锐湿疣组织中阳性表达率13．3％（4／30），正常组织无阳性表达（0／10），两者之间无显著性差异（P〉0．05）；尖锐湿疣组织中PCNA与P21表达之间无相关性（r=0．196，P〉0．05）。结论：PCNA、P21过表达与尖锐湿疣角质形成细胞自限性增殖密切相关。     

男性尖锐湿疣三种疗法的临床观察

目的探讨减少尖锐湿疣复发的有效方法。方法分别用包皮环切术加二氧化碳激光（甲组）治疗、二氧化碳激光治疗（乙组）和液氧（-196℃）治疗（丙组）男性尖锐湿疣，并做疗效对比和复发率观察。结果甲组治愈率90.4％，乙组治愈率57.6％，丙组治愈率31．8％，甲组与乙、丙组疗效比较差异在统计学上均有非常显著性（P〈0．001），乙、丙组疔效比较差异无显著性（P〉0．05）。追踪观察9个月末三组复发率，甲组10．6％，乙组41．3％，丙组47．7％，差异有非常显著性（P〈0．001）。乙、丙组复发率比较差异无显著性（P〉0．05）。结论包皮环切术加二氧化碳激光治疗男性尖锐湿疣可以除去尖锐湿疣临床表现的病灶，改变闭合潮湿环境，减少尖锐湿疣的复发机会，且方法简便有效，治疗成本低，安全性高，无特殊不良反应，是临床上根治尖锐湿疣有效的治疗方法。     

MCP-1mRNA及其蛋白在尖锐湿疣表皮中的表达

目的：从mRNA和蛋白水平检测尖锐湿疣表皮中单核细胞趋化蛋白-1（MCP-1）的表达，探讨其在尖锐湿疣皮损局部免疫异常机制的作用。方法：采用原位杂交、免疫组化和Western免疫印迹的方法，检测30例尖锐湿疣表皮及20例正常包皮表皮中MCP-1mRNA及其蛋白的表达水平。结果：MCP-1mRNA及其蛋白表达部位主要在表皮基底层。MCP-1mRNA在尖锐湿疣皮损组和对照组中原位杂交染色阳性单位值分别为10．7±5．1和11．2±4．6，差异无统计学意义（t=-8．70，P〉0．05）；MCP-1蛋白在尖锐湿疣皮损组和对照组中免疫组化染色阳性单位值分别为10．9±5．1和11．5±4．9，差异无统计学意义（t=-0．981，P〉0．05）；Western免疫检测结果显示尖锐湿疣皮损组和对照组表达的结合珠蛋白平均光密度值分别为0．18±0．06和0．20±0．06，两者差异无统计学意义（t=-0．721，P〉0．05）。结论：尖锐湿疣患者皮损MCP-1无论是其mRNA水平还是蛋白水平均表达微弱，可能是导致尖锐湿疣免疫微环境异常的原因之-。     

综合治疗女性尖锐湿疣的临床效果观察

目的：探讨女性尖锐湿疣综合治疗的临床效果。方法：我院治疗女性尖锐湿疣患者80例随机分为两组：观察组40例进行综合治疗，包括物理治疗、抗病毒治疗、抗茵治疗、中药消疣汤熏蒸治疗；对照组40例行常规物理治疗。观察并对比两组患者治愈效果和复发情况。结果：观察组与对照组治愈率分别为77．5％、40．0％，相比具有显著性差异（P〈0．01）；观察组治疗总有效率97．5％显著高于对照组的75．0％（P〈O．05）；治疗3—6个月内观察组复发率为5．1％，显著低于对照组的23．3％（P〈0．05）。结论：综合法治疗女性尖锐湿疣治愈率高，治疗效果好，复发率低，值得临床推广。     

中药（消疣汤）对预防尖锐湿疣复发初探

目的：评价消疣汤治疗尖锐湿疣的疗效及其免疫调节作用。方法：62例患者分为中药治疗组和阿昔洛韦对照组,治疗30天后跟踪观察半年,治疗前后采用双抗体夹心ELISA法检测血清IL－2。结果：治疗组和对照组临床疗效基本相同（P＞0．05）,治疗前后血清IL－2水平,治疗组明显升高（P＜0．05）,对照组无明显差异（P＞0．05）。结论：中药消疣汤是治疗尖锐湿疣的有效药物,它不仅具有抗病毒作用而且具有免疫调节作用,对减少尖锐湿疣的复发有一定作用。     

自体疣埋植治疗尖锐湿疣及其对机体T细胞亚群的影响

目的：研究自体疣组织埋植治疗尖锐湿疣（CA）疗效及其对机体细胞免疫功能的影响。方法：将98例尖锐湿疣患者随意分成两组，A组采用自体疣组织埋植＋CO2激光治疗，B组单用CO2激光治疗；两组治疗前后均检测外周血T淋巴细胞亚群。结果：A组复发率低，疗效优于B组，A组与B组比较，CD8^ 细胞数量明显下降（P＜0．05），CD4^ 细胞数量及CD4^ /CD8^ 比值明显提高（P＜0．05）。结论：自体疣组织埋植治疗尖锐湿疣疗效好，能降低复发率，其机理可能是自体疣组织埋植能改善CA患者的细胞免疫功能。     

祛疣灵联合电离子治疗尖锐湿疣疗效观察及对细胞增殖与凋亡的影响

我们应用“祛疣灵”外洗联合电离子治疗66例尖锐湿疣,并采用免疫组织化学和TUNEL法对15例患者中药外洗前后分别进行增殖细胞核抗原(PCNA)及凋亡细胞表达影响的研究。中药“祛疣灵”联合电离子治疗尖锐湿疣临床治愈率为81.8%;PCNA在中药外洗2周后较外洗前表达明显减少,而凋亡细胞较疗前明显增多,其差异有统计学意义。“祛疣灵”是治疗尖锐湿疣的有效外用方剂,其作用机制可能是通过抑制疣体细胞增殖,并促进细胞凋亡,达到治疗尖锐湿疣的目的。     

表皮生长因子受体在尖锐湿疣皮损中的表达

为探讨表皮生长因子受体（EGFR）在尖锐湿疣（CA）细胞增殖中的调节作用。用免疫组化SP法检测EGFR在56例尖锐湿疣与30名正常皮肤中的表达。结果：EGFR在尖锐湿疣组织除角质层外的表皮全层细胞均有强表达，正常皮肤中EGFR也有表达，但主要在基底层细胞表达，EGFR在尖锐湿疣皮损颗粒层细胞和棘层细胞中的表达显著强于正常皮肤（P＜0．01），结果示尖锐湿疣皮损中EGFR的表达增强可能与HPV6／11型感染上皮细胞过度增殖的致病过程有关。     

P53 PCNA K-i67 bcl-2在尖锐湿疣中的表达

目的　探讨尖锐湿疣 (CA)中P5 3、Ki 6 7、bcl 2、增殖细胞核抗原 (PCNA)表达与人乳头瘤病毒 (HPV )感染的关系。方法　用免疫组化方法对 40例CA和 10例正常包皮进行检测。结果　 40例CA中P5 3、Ki 6 7、bcl 2、PCNA阳性标本分别为 33例 ( 82 .5 % )、37例 ( 92 .5 % )、2 0例 ( 5 0 .0 % )、35例 ( 87.5 % )。CA组各种蛋白的表达强度均明显高于对照组 (P <0 .0 5 )。HPV感染的空泡细胞核中同时有P5 3、Ki 6 7、PCNA过度表达者 2 5例。P5 3、bcl 2阳性细胞多分布于基底层、棘细胞中下层。结论　CA中存在P5 3、Ki 6 7、PCNA、bcl 2过度表达 ,提示角质形成细胞异常增生与HPV感染有关。     

Evidence of increased apoptosis and reduced proliferation in basal cell carcinomas treated with tazarotene

A preliminary clinical experience suggested tazarotene, a new acetylenic retinoid, as an effective alternative topical treatment of basal cell carcinomas (BCC). The mechanisms of action of this synthetic retinoid, however, have not been yet clarified. In this work we assessed the in vivo effects of daily application of tazarotene for 24 wk, on 30 small superficial and nodular BCC, and the in vitro effects of tazarotene on immortalized basal and squamous tumor epidermal cells. Cellular proliferation, apoptosis and changes in expression of retinol and retinoic acid receptors (RAR), p53, bcl-2, and bax were studied by immunohistochemistry, western blotting and PCR. Overall, 76.7% of treated tumors showed >50% regression. Complete healing was observed in 46.7% of all treated BCC, without recurrences at 2-y observation. Regression was associated with reduced proliferation and increased apoptosis, demonstrated by Ki-67- and TdT-mediated dUTP-biotin nick-end labelling-positive nuclear staining, and with enhanced RAR-beta and bax expression, with RAR-alpha and -gamma expression unchanged. In vitro, tazarotene induced a concentration-dependent increase of RAR-beta and bax associated with a greater rate of apoptosis and growth inhibition in basaloid tumor cells compared with squamous tumor cells. Our studies provide convincing evidence that tazarotene induces BCC regression possibly by synergistic RAR-beta-dependent anti-proliferative and pro-apoptotic pathways.     

二氧化碳激光联合重组人干扰素α-2b治疗60例尖锐湿疣患者的临床观察

目的：观察尖锐湿疣（CA）应用二氧化碳激光联合重组人干扰素α-2b治疗的有效性、安全性、复发率。方法：将筛选的120例尖锐湿疣患者随机分为试验组和对照组。试验组和对照组患者均行二氧化碳激光术局部治疗。试验组60例患者加用重组人干扰素α-2b皮损局部注射。观察两组的治愈率、复发率。结果：试验组术后治愈率为91．67％,随访复发率为8．33％。对照组治愈率为68．34％,随访复发率为31．67％。两组间治愈率、复发率比较,均有显著性统计学差异（P〈0．01）。结论：应用二氧化碳激光联合重组人干扰素α-2b局部注射对尖锐湿疣疗效较好,并且术后复发率较低。     

Expression of bcl-2, p53 and Ki-67 in arsenical skin cancers

To investigate the regulation of apoptosis and proliferation in arsenic-induced skin cancers, we examined the expression of bcl-2. p53, and Ki-67 using immunohistochemical staining. Thirty patients with Bowen's disease (BD), ten with basal cell carcinoma (BCC), eight with squamous cell carcinoma (SCC) and eleven of perilesional normal skin (PLN) of the non-sun exposure sites from endemic area were examined. The results showed that: 1) bcl-2 was expressed in all of the BCC homogeneously, in none of the SCC, and in 12/30 of the BD focally or homogeneously; 2) p53 was expressed in all of the arsenical skin cancers with a labelling index of 75±14% of BD, 50±17% of BCC. 61±15% of SCC, and also in all of the perilesional normal skin with a labelling index of 55±24%; 3) Ki-67 was expressed in all of the skin cancers with labelling index of 58±17% of BD. 12±7% of BCC, 47±21% of SCC, and in 9/11 of PLN with a labelling index of 41±24%. Expression of bcl-2 in BCC or BD is related to the phenotype of germinative basal cell. The constant expression of bcl-2 i early dysplastic cells of BD and the earliest expression of P53 in the basal cells of perilesional normal skin indicate that the initial step of arsenic-induced carcinogenesis is from the basal germinative cells. There is no mutual relationship between bcl-2, p53 or Ki-67 expression in any type of the arsenical skin cancers, but there is a positive correlation between p53 and Ki-67 expression identified in perilesional normal skin. BD had the highest labelling index of p53 and Ki-67.     

角化棘皮瘤与高分化鳞状细胞癌细胞凋亡及凋亡基因bcl-2和bax比较研究

目的：探讨角化棘皮瘤与皮肤高分化鳞癌在细胞凋亡方面的差异。方法：用脱氧核苷酰转移酶介导的ｄ－ＵＴＰ生物素缺口末端标记技术，原位检测了ＫＡ和ｗＳＣＣ的凋亡细胞。用免疫组化研究技术研究了皮损部位与细胞凋亡有关ｂａｘ和ｂｃｌ－２的基因产物的表达。结果：凋亡细胞在ＫＡ发生率为８０％；在ｗＳＣＣ也为８０％。     

尖锐湿疣组织中的细胞凋亡与caspase-3和bcl-2蛋白的检测

目的探讨尖锐湿疣(CA)细胞凋亡与caspase-3和bcl-2蛋白的水平及其相互关系。方法对于33例CA,5例巨大CA患者皮损和8例正常皮肤组织用TUNEL法原位检测角质形成细胞(KC)的凋亡,用免疫组化法检测caspase-3和bcl-2蛋白的水平。结果33例CA患者中30例(90.91%)凋亡指数(AI)阳性,5例巨大CA患者的AI均为阳性。33例CA和5例巨大CA患者caspase-3蛋白均为阳性。Bcl-2在CA和巨大CA的阳性率分别为30.30%和20.00%。正常对照的AI及caspase-3和bcl-2蛋白均为阴性。在CA和巨大CA两组之间,AI、caspase-3及bcl-2的阳性率均无显著性差异(P>0.05)。CA组的AI与caspase-3蛋白的水平呈极显著正相关(P<0.0001),与bcl-2的水平呈极显著负相关(P<0.0001),caspase-3与bcl-2的水平呈极显著负相关(P<0.0001)。结论CA皮损组织的KC中存在着明显的细胞凋亡现象,caspase-3和bcl-2可能参与了CA细胞凋亡的调节机制。     

Notch1基因对裸鼠皮肤鳞状细胞癌移植瘤细胞增殖和凋亡的影响

目的 探讨Notch1基因在人皮肤鳞状细胞癌SCL-1细胞裸鼠移植瘤中的作用。方法 于裸鼠腋下接种0.2 ml 1 × 108/ml的SCL-1细胞悬液,建立人皮肤鳞状细胞癌裸鼠移植瘤模型,将其分为三组：未处理组（接种PBS悬浮的未转染细胞）,空载体转染组（接种空载体转染的细胞）和Notch1转染组（接种Notch1转染的细胞）。连续2周观察转染荷瘤裸鼠的生长情况,采用TdT介导的dUTP缺口末端标记技术（TUNEL）研究肿瘤组织中的细胞凋亡,利用RT-PCR和Western印迹研究肿瘤组织中Notch1、bcl-2和bax mRNA和蛋白的表达。结果 Notch1转染组中肿瘤的生长明显受到抑制,其肿瘤的重量为（0.574 ± 0.219） g,显著低于未处理组（2.642 ± 0.404） g和空载体转染组（2.606 ± 0.512） g（F = 26.642,P 值均 < 0.01）。TUNEL结果表明,Notch1转染组每500个细胞中凋亡的细胞数为87 ± 9,明显高于未处理组（8 ± 2）和空载体转染组（10 ± 3）中细胞凋亡的数目（P < 0.05）;此外,RT-PCR和Western印迹结果表明Notch1转染组中裸鼠肿瘤组织Notch1和bax mRNA和蛋白表达均显著上升,而bcl-2的表达显著下调,三组间比较差异具有统计学意义（P值均 ＜ 0.05）。结论 Notch1基因能抑制人皮肤鳞状细胞癌细胞SCL-1裸鼠皮下移植瘤的生长,诱导SCL-1细胞凋亡,后者可能与bax表达的上升和bcl-2表达的下调相关。     

Immunohistological detection of proliferating cells in normal and psoriatic epidermis using Ki-67 monoclonal antibody

Ki-67 monoclonal antibody (mAb) has been suggested to react only with proliferating cells. In order to study epidermal proliferating cells, immunohistological staining was performed with Ki-67 on 14 skin specimens from normal subjects and 30 from patients suffering from psoriasis. Staining of nuclei by Ki-67 was clearly observed in the epidermis. In normal epidermis, Ki-67+ cells were sparse in the basal layer (L1) and next upper layer (L2), while in psoriatic epidermis, they were abundant in L1, L2 and a few layers above L2. The percentage of positive cells in L1 of normal and psoriatic skin were 4.5 and 54%, respectively. Double staining (Ki-67 and S phase stain using bromodeoxyuridine) was also performed, and the results showed that: 1) the distribution patterns of Ki-67+ cells and those of S phase cells were similar in every section examined; 2) Ki-67 stained almost all S phase cells, and 3) Ki-67 also recognized considerable numbers of non-S phase cells. Therefore our data indicate that Ki-67 can detect a certain population of epidermal proliferating cells that includes S phase cells. Although it remains unclear whether all the proliferating cells in the epidermis can be detected by this mAb, we suggest that Ki-67 staining is an easy and useful technique for evaluating the proliferative activity of the epidermis.     

Immunohistochemical characterization of pleomorphic giant cells in basal cell carcinoma

Over 20 cases of basal cell carcinomas with pleomorphic giant cells of the mononuclear or multinucleate type have been described. The nature of these cellular and nuclear changes has not been elucidated. Some authors found that these cells have phagocytic properties and others reported an aneuploid DNA content. We have seen mitotic figures in some of these giant cells, and postulate that these cells are capable of proliferation. Twelve cases of basal cell carcinoma with pleomorphic giant cells were examined using monoclonal antibodies recognizing the proliferating cell nuclear antigen (PCNA), Ki-67, and bcl-2 antigens. Expression of proliferation associated antigens in the giant cell population was higher than in the small cell population. Over expression of bcl-2 was detected in both the small and giant cells in all cases. The results demonstrate that the giant tumor cells are cycling and express bcl-2 protein in a manner consistent with basal cell carcinoma. The changes are unlikely to represent a senescent change as seen occasionally in mesenchymal neoplasms.