超微结构及免疫组化染色下观察藓化饮对金黄地鼠颊黏膜癌前病变的逆转作用
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摘要
目的:口腔黏膜癌前病变与口腔癌的发生密切相关,近些年来,为了降低口腔癌的发病率,人们越来越重视对口腔黏膜癌前病变的预防和治疗。目前,口腔黏膜癌前病变的治疗手段较多,主要分为药物治疗和非药物治疗。非药物治疗如手术治疗、激光治疗、冷冻治疗等等,有着适应证的局限性。而西药治疗如:增生平、维甲酸等等,长期服用可能会出现一些毒副作用。近年来研究表明,中医中药治疗口腔黏膜癌前病变显示出明显的临床效果,也成为目前口腔黏膜癌前病变研究的热点。本课题通过建立口腔黏膜癌前病变动物模型,应用电镜和免疫组化的方法观察中药藓化饮对口腔黏膜癌前病变的逆转作用。
方法:选取SPF级LVG叙利亚金黄地鼠55只,鼠龄六周,体质量80g。按完全随机设计方法将动物分为:空白对照组5只和实验组50只。实验组用于建立口腔黏膜癌前病变动物模型,其中又分为癌前病变对照组10只、藓化饮治疗组30只、生理盐水治疗对照组10只。将实验组50只动物,颊囊黏膜涂以0.5%DMBA丙酮溶液,每周3次,连续涂药6周后,藓化饮治疗组、生理盐水治疗对照组分别灌胃治疗。治疗8周时,各组同步取材。采用电镜及免疫组化方法,观察中药藓化饮对金黄地鼠颊黏膜癌前病变的逆转作用。
结果:
1癌前病变动物模型的建立
实验组50只动物颊黏膜致癌六周后,双侧颊黏膜肉眼观全部为充血、糜烂、条索状皱折等表现。随机切取10例不同肉眼表现的部分病变部位,HE染色后观察,上皮黏膜表现为不同程度的异常增生。由此说明,经DMBA丙酮溶液致癌六周后,已成功建立金黄地鼠颊黏膜癌前病变的动物模型。
2藓化饮对叙利亚金黄地鼠口腔黏膜癌前病变的逆转作用
2.1肉眼观察
空白对照组金黄地鼠颊黏膜表面光滑,呈淡粉色,透亮具有弹性,血管清晰。实验组50只动物涂DMBA丙酮溶液六周后,颊黏膜变得粘连、粗糙、无弹性、充血糜烂,甚至可见条索状皱折。中药藓化饮灌胃八周后,可见颊黏膜皱折减轻,黏膜表面逐渐光滑,充血糜烂面基本消失,但弹性较正常稍差。生理盐水灌胃治疗八周后与治疗前无明显改变。
2.2光镜观察
空白对照组金黄地鼠颊黏膜为角化的复层鳞状上皮,上皮和肌层之间为薄层结缔组织及少量散在的纤维细胞和血管。停涂DMBA丙酮溶液八周后,镜下观察金黄地鼠颊黏膜可见不同程度的异常增生。中药藓化饮灌胃治疗八周后,黏膜上皮层次逐渐清晰,上皮层数逐渐恢复正常,基底层细胞排列比较有规则,异型性的细胞减少。生理盐水灌胃治疗八周后与治疗前无明显改变。
2.3扫描电镜观察
空白对照组金黄地鼠颊黏膜上皮表现为多边形细胞,细胞间桥很明显,个体细胞表面显示很多网状排列的微嵴,微嵴围成均匀一致的小窝,整个细胞表面呈现一种蜂窝状外观。停涂DMBA丙酮溶液八周,细胞外形更不规则,细胞重叠,松解现象更为突出,微嵴肿胀变宽,不规则排列,呈卵石状外观,蜂窝状结构完全消失。中药藓化饮灌胃治疗八周后,细胞外形逐渐清晰,呈现出多边形。细胞表面大部分区域呈现蜂窝状结构,可见连续的细胞界限。生理盐水灌胃治疗八周与治疗前无明显改变。
2.4透射电镜观察
空白对照组金黄地鼠颊黏膜上皮细胞外形规则,细胞紧密相邻,桥粒丰富,张力纤维发育正常,基底膜完整。停涂DMBA丙酮溶液八周后,可见细胞间隙增宽,桥粒大量减少,张力纤维减少,细胞外形不规则,核仁边集,核外形不规则,核膜内陷,可见基底膜断裂。中药藓化饮灌胃治疗八周后,增宽的细胞间隙逐渐减小,桥粒和张力纤维逐渐丰富,细胞外形逐渐规则。生理盐水灌胃治疗八周与治疗前无明显改变。
3藓化饮治疗后EZH2、PCNA在各组免疫组化染色结果
EZH2、PCNA在空白对照组和藓化饮治疗组低表达,而在癌前病变对照组和生理盐水治疗对照组高表达。癌前病变对照组EZH2、PCNA高表达率与空白对照组相比,有显著性差异(P<0.05);藓化饮治疗组EZH2、PCNA高表达率与癌前病变对照组相比,有显著性差异(P<0.05);而藓化饮治疗组EZH2、PCNA高表达率与空白对照组相比,无显著性差异(P>0.05);生理盐水治疗对照组EZH2、PCNA高表达率与藓化饮治疗组相比,有显著性差异(P<0.05)。
统计学分析,两指标表达具有相关性,表达吻合度具有统计学意义。
结论:
1使用0.5%DMBA丙酮溶液涂抹叙利亚金黄地鼠颊黏膜六周后,能成功建立金黄地鼠颊黏膜癌前病变动物模型。
2藓化饮治疗八周后,叙利亚金黄地鼠颊黏膜充血糜烂面基本消失,细胞外形逐渐清晰,上皮层异型性的细胞减少,EZH2、PCNA高表达率明显下降。说明藓化饮对金黄地鼠颊黏膜癌前病变有明显逆转作用。
Objective:There was a correlation between oral mucosal premalignant lesion and carcinoma of oral mucosa. These years, people had been paid more and more attention on the carcinoma of oral mucosa preservation and treatment to cut down the disease incidence of carcinoma of oral mucosa. Now, there were lots of therapeutic tool for oral mucosal premalignant lesion. And mainly divide into drug treatment and un-drug treatment. The un-drug treatment usually included operation treatment、laser therapy、freezing therapy and so on. But these ways had indication confine and higher recurrence of post-treatment, especially could not prevent the new lesion appearance in other part of oral cavity. So these defects greatly cut down the un-drug treatment possibility. Medical doctor treat such as ratinoic acid would had adverse reaction if long-term using. So, the emphasis of oral mucosal premalignant lesion drug treatment was to prevent the new lesion appearance in other part of oral cavity and deeply developed. And Chinese herbal medicine properly had these ideal condition. These years research showed that Chinese herbal medicine had played markedly therapeutic effect during the oral mucosal premalignant lesion treatment process. And it had become to oral mucosal premalignant lesion research hot spot. The topic investigated the role of Xianhuayin on the reversal of the premalignant lesion in the mucosa of cheek pouch in golden hamster by using immunohistochemical methodand electron microscope by premalignant lesion animal model generation.
Methods:A total of 55 SPF LVG Syrian golden hamsters (6 weeks old, weight: 80 g) were randomly divided into untreated control group (n=5) and experimental group (n=50). Animals in the experimental group were used to generate the oral mucosal premalignant lesion models. Of these 50 animals with oral mucosal premalignant lesions, 10 hamsters were treated with normal saline (NS-treated group); 10 hamsters were untreated (premalignant lesion group); 30 hamsters were treated with Xianhuayin (Xianhuayin-treated group). The buccal mucosa of 50 hamsters in the experimental group was painted with DMBA (0.5% in acetone) as described previously. Application with DMBA was performed three times a week for a continuous of 6 weeks. Then intragastic administration was performed once a day for a continuous 8 weeks in Xianhuayin-treated group and NS-treated group. All the animals were sacrificed and the tissue samples were collected at the same time. To investigate the role of Xianhuayin on the reversal of the premalignant lesion in the mucosa of cheek pouch in golden hamster by using immunohistochemical method、electron microscope.
Results:
1 Premalignant lesion animal model generation
Six weeks after treatment with DMBA acetone solution, in the experimental group, both sides of the cheek pouch mucosa of hamsters became adhered, lacking of elasticity, congested, anabrotic and streak-like ruffle. Histological analysis was performed by randomly collecting ten part of lesion tissues, which had different of visual manifestation. Variable degree of epithelial dysplasia was observed under light microscopic . So six weeks after treatment with DMBA acetone solution, it had successfully generated the premalignant lesion animal model.
2 Role of Xianhuayin on the reversal of premalignant lesion in the oral buccal mucosa of golden hamsters
2.1 Visual observations
The cheek pouch mucosa of hamsters in the untreated-group exhibited smooth surface, light pink color, transparency, elasticity and clear blood vessels. Six weeks after treatment with DMBA acetone solution, the cheek pouch mucosa of hamsters in the experimental group became adhered, rough, lacking of elasticity, congestion, anabrosis and streak-like ruffle was evident . Eight weeks after intragastric administration with Xianhuayin, the adherence of cheek pouch mucosa became mitigated, smooth surface appeared gradually, and congestion as well as anabrosis disappeared, but the elasticity was still relatively poorer than the normal tissues. No significant changes were observed after the hamsters in the premalignant lesion group were intragastrically administered with normal saline .
2.2 Light microscopic observations
Buccal mucosa of the normal hamsters in the untreated group was keratinized and stratified squamous epithelium under light microscope. Lamellar connective tissues, small amount of sporadic fibrocytes and blood vessels were present between epithelium and muscular layer . Variable degree of epithelial dysplasia were observed in the hamster of premalignant group 8 weeks after painting with DMBA was stopped. Eight weeks after treatment with Xianhuayin, the irregular epithelial mucosa gradually became distinct, the layer of epithelium was recovered to normality, basilar membrane cells were regularly organized and the number of atypical cells was reduced . No significant changes were observed after hamsters were intragastrically administered with normal saline .
2.3 SEM analysis
Scanning Electronic Microscopic (SEM) analysis showed that in the hamster of untreated-group, the buccal mucosa contained polygonal cells and clear cell bridge. The surface of the individual cells exhibited honeycomb structures . Eight weeks after painting with DMBA was stopped, the epithelial cells were morphologically irregular, overlapped and loosened. Swelling and widening micro-crests were irregularly arranged and appeared as gravel shape. Eight weeks after intragastric administration with Xianhuayin, most of the cell surface exhibited honeycomb structures and continuous cellular border was observed in the hamsters of Xianhuayin-treated group . No significant changes were observed for the hamsters in NS-treated group compared with those in the premalignant group.
2.4 TEM analysis Transmission Electronic Microscopic (TEM) analysis showed that in the hamsters of untreated group, buccal mucosal epithelial cells were morphologically regular, closely connected and contained abundant desmosomes. The tonofibrils were developed normally. Eight weeks after painting with DMBA was stopped, desmosomes and tonofibrils were significantly reduced, intercellular gap was increased, the shape of epithelial cells were irregular, the morphologically irregular nucleolus was marginally gathered and the caryotheca was invaginated. Eight weeks after intragastric administration with Xianhuayin, the widening intercellular gap became reduced gradually, desmosomes and tonofibrils were gradually becoming abundant and the cells were becoming morphologically regular. No significant difference was observed between the hamsters in NS-treated group and those in the premalignant group.
3 Expression of EZH2、PCNA in different group after Xianhuayin treatment
EZH2、PCNA were rarely expressed or lower expressed in the untreated group and Xianhuayin-treated group, and they were higher expressed in the premalignant group and NS-treated group.
The rate of EZH2、PCNA high expression of the premalignant lesion group compared with the untreated control group’s, it had statistical difference (P<0.05); The rate of EZH2、PCNA high expression of the Xianhuayin-treated group compared with the premalignant lesion group’s, it had statistical difference (P<0.05); The rate of EZH2、PCNA high expression of the Xianhuayin-treated group compared with the untreated control group’s, it had no statistical difference (P>0.05); The rate of EZH2、PCNA high expression of the NS-treated group compared with the Xianhuayin-treated group’s, it had statistical difference (P<0.05)
Statistical analysis, there was a correlation between EZH2 and PCNA, and the expression goodness of fit had statistical difference.
Conclusion:
1 Six weeks after treatment with DMBA acetone solution, it had successfully generated the premalignant lesion animal model.
2 Eight weeks after intragastric administration with Xianhuayin, congestion as well as anabrosis disappeared; the cells were becoming morphologically regular; the number of atypical cells was reduced; the rate of EZH2、PCNA high expression obviously decreased. It’s concluded that Xianhuayin had the obvious role on the reversal of premalignant mucosal lesions in the golden hamster buccal pouch.
方法:选取SPF级LVG叙利亚金黄地鼠55只,鼠龄六周,体质量80g。按完全随机设计方法将动物分为:空白对照组5只和实验组50只。实验组用于建立口腔黏膜癌前病变动物模型,其中又分为癌前病变对照组10只、藓化饮治疗组30只、生理盐水治疗对照组10只。将实验组50只动物,颊囊黏膜涂以0.5%DMBA丙酮溶液,每周3次,连续涂药6周后,藓化饮治疗组、生理盐水治疗对照组分别灌胃治疗。治疗8周时,各组同步取材。采用电镜及免疫组化方法,观察中药藓化饮对金黄地鼠颊黏膜癌前病变的逆转作用。
结果:
1癌前病变动物模型的建立
实验组50只动物颊黏膜致癌六周后,双侧颊黏膜肉眼观全部为充血、糜烂、条索状皱折等表现。随机切取10例不同肉眼表现的部分病变部位,HE染色后观察,上皮黏膜表现为不同程度的异常增生。由此说明,经DMBA丙酮溶液致癌六周后,已成功建立金黄地鼠颊黏膜癌前病变的动物模型。
2藓化饮对叙利亚金黄地鼠口腔黏膜癌前病变的逆转作用
2.1肉眼观察
空白对照组金黄地鼠颊黏膜表面光滑,呈淡粉色,透亮具有弹性,血管清晰。实验组50只动物涂DMBA丙酮溶液六周后,颊黏膜变得粘连、粗糙、无弹性、充血糜烂,甚至可见条索状皱折。中药藓化饮灌胃八周后,可见颊黏膜皱折减轻,黏膜表面逐渐光滑,充血糜烂面基本消失,但弹性较正常稍差。生理盐水灌胃治疗八周后与治疗前无明显改变。
2.2光镜观察
空白对照组金黄地鼠颊黏膜为角化的复层鳞状上皮,上皮和肌层之间为薄层结缔组织及少量散在的纤维细胞和血管。停涂DMBA丙酮溶液八周后,镜下观察金黄地鼠颊黏膜可见不同程度的异常增生。中药藓化饮灌胃治疗八周后,黏膜上皮层次逐渐清晰,上皮层数逐渐恢复正常,基底层细胞排列比较有规则,异型性的细胞减少。生理盐水灌胃治疗八周后与治疗前无明显改变。
2.3扫描电镜观察
空白对照组金黄地鼠颊黏膜上皮表现为多边形细胞,细胞间桥很明显,个体细胞表面显示很多网状排列的微嵴,微嵴围成均匀一致的小窝,整个细胞表面呈现一种蜂窝状外观。停涂DMBA丙酮溶液八周,细胞外形更不规则,细胞重叠,松解现象更为突出,微嵴肿胀变宽,不规则排列,呈卵石状外观,蜂窝状结构完全消失。中药藓化饮灌胃治疗八周后,细胞外形逐渐清晰,呈现出多边形。细胞表面大部分区域呈现蜂窝状结构,可见连续的细胞界限。生理盐水灌胃治疗八周与治疗前无明显改变。
2.4透射电镜观察
空白对照组金黄地鼠颊黏膜上皮细胞外形规则,细胞紧密相邻,桥粒丰富,张力纤维发育正常,基底膜完整。停涂DMBA丙酮溶液八周后,可见细胞间隙增宽,桥粒大量减少,张力纤维减少,细胞外形不规则,核仁边集,核外形不规则,核膜内陷,可见基底膜断裂。中药藓化饮灌胃治疗八周后,增宽的细胞间隙逐渐减小,桥粒和张力纤维逐渐丰富,细胞外形逐渐规则。生理盐水灌胃治疗八周与治疗前无明显改变。
3藓化饮治疗后EZH2、PCNA在各组免疫组化染色结果
EZH2、PCNA在空白对照组和藓化饮治疗组低表达,而在癌前病变对照组和生理盐水治疗对照组高表达。癌前病变对照组EZH2、PCNA高表达率与空白对照组相比,有显著性差异(P<0.05);藓化饮治疗组EZH2、PCNA高表达率与癌前病变对照组相比,有显著性差异(P<0.05);而藓化饮治疗组EZH2、PCNA高表达率与空白对照组相比,无显著性差异(P>0.05);生理盐水治疗对照组EZH2、PCNA高表达率与藓化饮治疗组相比,有显著性差异(P<0.05)。
统计学分析,两指标表达具有相关性,表达吻合度具有统计学意义。
结论:
1使用0.5%DMBA丙酮溶液涂抹叙利亚金黄地鼠颊黏膜六周后,能成功建立金黄地鼠颊黏膜癌前病变动物模型。
2藓化饮治疗八周后,叙利亚金黄地鼠颊黏膜充血糜烂面基本消失,细胞外形逐渐清晰,上皮层异型性的细胞减少,EZH2、PCNA高表达率明显下降。说明藓化饮对金黄地鼠颊黏膜癌前病变有明显逆转作用。
Objective:There was a correlation between oral mucosal premalignant lesion and carcinoma of oral mucosa. These years, people had been paid more and more attention on the carcinoma of oral mucosa preservation and treatment to cut down the disease incidence of carcinoma of oral mucosa. Now, there were lots of therapeutic tool for oral mucosal premalignant lesion. And mainly divide into drug treatment and un-drug treatment. The un-drug treatment usually included operation treatment、laser therapy、freezing therapy and so on. But these ways had indication confine and higher recurrence of post-treatment, especially could not prevent the new lesion appearance in other part of oral cavity. So these defects greatly cut down the un-drug treatment possibility. Medical doctor treat such as ratinoic acid would had adverse reaction if long-term using. So, the emphasis of oral mucosal premalignant lesion drug treatment was to prevent the new lesion appearance in other part of oral cavity and deeply developed. And Chinese herbal medicine properly had these ideal condition. These years research showed that Chinese herbal medicine had played markedly therapeutic effect during the oral mucosal premalignant lesion treatment process. And it had become to oral mucosal premalignant lesion research hot spot. The topic investigated the role of Xianhuayin on the reversal of the premalignant lesion in the mucosa of cheek pouch in golden hamster by using immunohistochemical methodand electron microscope by premalignant lesion animal model generation.
Methods:A total of 55 SPF LVG Syrian golden hamsters (6 weeks old, weight: 80 g) were randomly divided into untreated control group (n=5) and experimental group (n=50). Animals in the experimental group were used to generate the oral mucosal premalignant lesion models. Of these 50 animals with oral mucosal premalignant lesions, 10 hamsters were treated with normal saline (NS-treated group); 10 hamsters were untreated (premalignant lesion group); 30 hamsters were treated with Xianhuayin (Xianhuayin-treated group). The buccal mucosa of 50 hamsters in the experimental group was painted with DMBA (0.5% in acetone) as described previously. Application with DMBA was performed three times a week for a continuous of 6 weeks. Then intragastic administration was performed once a day for a continuous 8 weeks in Xianhuayin-treated group and NS-treated group. All the animals were sacrificed and the tissue samples were collected at the same time. To investigate the role of Xianhuayin on the reversal of the premalignant lesion in the mucosa of cheek pouch in golden hamster by using immunohistochemical method、electron microscope.
Results:
1 Premalignant lesion animal model generation
Six weeks after treatment with DMBA acetone solution, in the experimental group, both sides of the cheek pouch mucosa of hamsters became adhered, lacking of elasticity, congested, anabrotic and streak-like ruffle. Histological analysis was performed by randomly collecting ten part of lesion tissues, which had different of visual manifestation. Variable degree of epithelial dysplasia was observed under light microscopic . So six weeks after treatment with DMBA acetone solution, it had successfully generated the premalignant lesion animal model.
2 Role of Xianhuayin on the reversal of premalignant lesion in the oral buccal mucosa of golden hamsters
2.1 Visual observations
The cheek pouch mucosa of hamsters in the untreated-group exhibited smooth surface, light pink color, transparency, elasticity and clear blood vessels. Six weeks after treatment with DMBA acetone solution, the cheek pouch mucosa of hamsters in the experimental group became adhered, rough, lacking of elasticity, congestion, anabrosis and streak-like ruffle was evident . Eight weeks after intragastric administration with Xianhuayin, the adherence of cheek pouch mucosa became mitigated, smooth surface appeared gradually, and congestion as well as anabrosis disappeared, but the elasticity was still relatively poorer than the normal tissues. No significant changes were observed after the hamsters in the premalignant lesion group were intragastrically administered with normal saline .
2.2 Light microscopic observations
Buccal mucosa of the normal hamsters in the untreated group was keratinized and stratified squamous epithelium under light microscope. Lamellar connective tissues, small amount of sporadic fibrocytes and blood vessels were present between epithelium and muscular layer . Variable degree of epithelial dysplasia were observed in the hamster of premalignant group 8 weeks after painting with DMBA was stopped. Eight weeks after treatment with Xianhuayin, the irregular epithelial mucosa gradually became distinct, the layer of epithelium was recovered to normality, basilar membrane cells were regularly organized and the number of atypical cells was reduced . No significant changes were observed after hamsters were intragastrically administered with normal saline .
2.3 SEM analysis
Scanning Electronic Microscopic (SEM) analysis showed that in the hamster of untreated-group, the buccal mucosa contained polygonal cells and clear cell bridge. The surface of the individual cells exhibited honeycomb structures . Eight weeks after painting with DMBA was stopped, the epithelial cells were morphologically irregular, overlapped and loosened. Swelling and widening micro-crests were irregularly arranged and appeared as gravel shape. Eight weeks after intragastric administration with Xianhuayin, most of the cell surface exhibited honeycomb structures and continuous cellular border was observed in the hamsters of Xianhuayin-treated group . No significant changes were observed for the hamsters in NS-treated group compared with those in the premalignant group.
2.4 TEM analysis Transmission Electronic Microscopic (TEM) analysis showed that in the hamsters of untreated group, buccal mucosal epithelial cells were morphologically regular, closely connected and contained abundant desmosomes. The tonofibrils were developed normally. Eight weeks after painting with DMBA was stopped, desmosomes and tonofibrils were significantly reduced, intercellular gap was increased, the shape of epithelial cells were irregular, the morphologically irregular nucleolus was marginally gathered and the caryotheca was invaginated. Eight weeks after intragastric administration with Xianhuayin, the widening intercellular gap became reduced gradually, desmosomes and tonofibrils were gradually becoming abundant and the cells were becoming morphologically regular. No significant difference was observed between the hamsters in NS-treated group and those in the premalignant group.
3 Expression of EZH2、PCNA in different group after Xianhuayin treatment
EZH2、PCNA were rarely expressed or lower expressed in the untreated group and Xianhuayin-treated group, and they were higher expressed in the premalignant group and NS-treated group.
The rate of EZH2、PCNA high expression of the premalignant lesion group compared with the untreated control group’s, it had statistical difference (P<0.05); The rate of EZH2、PCNA high expression of the Xianhuayin-treated group compared with the premalignant lesion group’s, it had statistical difference (P<0.05); The rate of EZH2、PCNA high expression of the Xianhuayin-treated group compared with the untreated control group’s, it had no statistical difference (P>0.05); The rate of EZH2、PCNA high expression of the NS-treated group compared with the Xianhuayin-treated group’s, it had statistical difference (P<0.05)
Statistical analysis, there was a correlation between EZH2 and PCNA, and the expression goodness of fit had statistical difference.
Conclusion:
1 Six weeks after treatment with DMBA acetone solution, it had successfully generated the premalignant lesion animal model.
2 Eight weeks after intragastric administration with Xianhuayin, congestion as well as anabrosis disappeared; the cells were becoming morphologically regular; the number of atypical cells was reduced; the rate of EZH2、PCNA high expression obviously decreased. It’s concluded that Xianhuayin had the obvious role on the reversal of premalignant mucosal lesions in the golden hamster buccal pouch.
引文
1高岩. WHO口腔黏膜癌前病变组织学新分类及其进展[J].中国实用口腔科杂志, 2008,1(12): 705-708
2周曾同.提高口腔黏膜癌前病变的诊疗质量[J].中华口腔医学杂志,2005,40(2):89-91
3刘伟,周曾同.口腔白斑的中医药治疗现状及研究进展[J].临床口腔医学杂志,2009,25(1):53-55
4 The Ministry of Science and Technology of the People's Republic of China. Guidance suggestion of caring laboratory animals.2006-09-30
5周曾同,娄佳宁.绞股蓝总甙对金地鼠颊囊白斑癌变过程中端粒酶活性化学预防作用研究[J].临床口腔医学杂志,2008,24(4):234-237
6李萍,葛化冰,孙正.酪氨酸激酶抑制剂GW2974对口腔鳞状细胞癌生长抑制作用的实验研究[J].中华口腔医学杂志,2007,42(7):432-435
7许彦枝,辛晓红,刘亚娴,等.藓化饮对口腔扁平苔藓患者激活调节因子表达的影响[J].山东中医,杂志,2009,28(1):13-14
8许良中,杨文涛.免疫组织化学反应结果的判定标准[J].中国癌症杂志,1996,6(4):229-231
9吴明利,王士杰,徐志彬,等.家兔颊黏膜鳞状上皮癌前病变模型制备及癌前病变逆转研究[J].实用癌症杂志,2005,20(1):5-7
10刘栋才,杨竹林,杨乐平.胆囊良恶性病变组织EZH2和PTEN表达及意义[J].中南大学学报,2008,33(7):618-622
11 Gil J, Bernard D, Peters G. Role of polycomb group proteins in stem cell self-renewal and cancer [J].DNA Cell Biol, 2005, 24(2):117- 125
12苗芳,李劲松.EZH2和Ki-67在胃癌中的表达及其临床意义[J].泰山医学院学报,2008,29(2):87-89
13黄优,王林,马俊青,等.大鼠生长发育过程中颅底软骨联合细胞增殖和凋亡的研究[J].口腔医学,2008,28(2):57-60
14李晓光,于肖鹏,赵海兰,等.PCNA在口腔鳞癌中的表达及其临床意义[J].泰山医学院学报,2005,26(1) :20-22
15 Bracken AP , Pasini D , Capra M, et al. EZH2 is downstream of the pRB- E2F pathway , essential for proliferation and amplified in cancer[J ] . EMBO , 2003 , 22 (20) : 5 323- 5 335
16王文斌,祁佩时.电子显微技术研究生物膜特性[J].工业安全与环保,2008,34(7):15-16
17范刚启,宋祥龙,王辉,等.活血化瘀治疗癌及癌前病变效应的两重性与血管生成的关系[J].中国中西医结合杂志,2003,23(8):624-626
18魏道智,宁书菊,林文雄.佩兰的研究进展[J].时珍国医国药,2007,18(7):1782-1783
19高贵峰,杜作鹏.浅谈蝉蜕在临床上的多功能应用[J].中华实用中西医结合杂志,2008,21(8):650-651
20许继文,付春梅.丹参的药理作用研究进展[J].医学综述,2006,12(23):1467-1468
21杨媛媛,周刚,马晓康,等.赤芍的研究进展[J].医药导报,2008,27(1):67-69
22许红兰.苦参的药理研究进展[J].现代医药卫生,2008,24(20):3066-3067
23郑利光,翟所迪.甘草锌膜对实验性口腔溃疡的治疗作用及黏膜刺激性研究[J].中国新药杂志,2006,15(8) :601-603
24刘庆林.赤芍的临床配伍应用浅析[J].中国新药杂志,2009,24(1):131-132
25王晓菲,金鸣.红花抗炎作用机制研究进展[J].中国新药杂志,2007,36(1) :51-53
26徐萍利,雷正荣.中医药防治胃癌前病变的研究进展[J].中外健康文摘,2008,5(1) : 1- 4
1周曾同.提高口腔黏膜癌前病变的诊疗质量[J].中华口腔医学杂志,2005,40(2):89-91
2刘伟,周曾同.口腔白斑的中医药治疗现状及研究进展[J].临床口腔医学杂志,2009,25(1):53-55
3 Bokor-Bratic M.Prevalence of oral leukoplakia. Med Pregl[J]. 2003, 56 (11-12): 552-555
4姜倩,祝磊,高黎.口腔白斑发病因素的初步探讨[J].河南医学研究,2006,15(4):363-365
5高岩. WHO口腔黏膜癌前病变组织学新分类及其进展[J].中国实用口腔科杂志,2008,1(12):705-708
6周曾同,钟文静,张水龙.微波对白斑上皮异常增生12项病理特征出现率的影响[J].临床口腔医学杂志,1998,14(3):163-165
7周曾同,张水龙,王英,等.金地鼠颊囊上皮异常增生12项病理特征的光镜分析[J].上海口腔医学,1997,6(1):32-35
8葛姝云,周曾同.口腔癌前病变生物标志物的研究进展[J].中华口腔医学杂志,2005,40(2):166-168
9孟宪瑞,刘进忠.口腔癌的早期诊断[J].国际口腔医学杂志,2008,35(3):329-331
10孙善珍.口腔癌前病变的诊断及其恶变潜能的监测[J].山东医药,1997,37(10):33
11周曾同,张水龙.口腔黏膜白斑研究概况[J].中华口腔医学杂志,1999,34(6):382-384
12庞劲凡.口腔白斑的药物治疗[J].武警医学,2008,19(4):349-352
13黄霞萍.益气活血、通络解毒法治疗口腔扁平苔藓30例[J].中医研究,2008,21(12):28-30
14王贵林.灯盏细辛注射液对不稳定心绞痛患者血流变学的影响[J].中国现代药物应用,2009,3(2):121-122
15娄佳宁,周曾同.绞股蓝总甙对金地鼠颊囊白斑癌变过程中端粒酶活性化学预防作用研究[J].临床口腔医学杂志,2008,24(4):234-236
16张为新,胡慧芳,李伟国,等.7种中药对实验性口腔癌阻断作用的研究[J].上海口腔医学,2004,13:34-37
17许彦枝,辛晓红,刘亚娴,等.藓化饮对口腔扁平苔藓患者激活调节因子表达的影响[J].山东中医杂志,2009,28(1):13-14
18张玉禄,李军祥,朱陵群.三种活血法对大鼠萎缩性胃炎癌前病变早期细胞凋亡的影响[J].中国中西医结合杂志,2008,28(5):448-450
19郑利光,翟所迪.甘草锌膜对实验性口腔溃疡的治疗作用及黏膜刺激性研究[J].中国新药杂志,2006,15(8):601-603
20陈谦明,李秉琦.口腔白斑病诊断、分级与治疗中的新观点[J].华西口腔医学杂志,2004,22(2):142-144
21林梅.口腔癌前病变的临床检查和风险评估[J].中华口腔医学研究杂志(电子版),2008,2(5):1-3
22周曾同.口腔白斑、红斑和黑斑的诊断与治疗[J].中华口腔医学杂志, 2006,41(8):502-505
23常云龙,左德崴.微波治疗舌下腺囊肿92例临床观察[J].中国医学创新,2008,5(35):52
24秦文敏,田永云.螺旋藻的药理研究进展[J].实用医技杂志,2007,14(20):2830-2831
25刘全未,黄维义.姜黄素抗动脉粥样硬化作用研究进展[J].心血管病学进展,2009,30(1):176-178
26许彦枝,李少成,王小玲.天然胡萝卜素对叙利亚金黄地鼠颊黏膜癌前病变逆转作用的实验研究[J].现代口腔医学杂志,2001,15(3):471,571
27尹鸿.灯盏细辛治疗冠心病心绞痛的疗效分析[J].中国医药指南,2008,6(17):344
28李芳,孙正,吴洪儒,等.Zyflamend对实验性口腔癌前病变化学预防作用的研究[J].北京口腔医学,2007,15(2):61-64
29陈伟,刘华一.中医药防治胃癌前病变机制的实验研究进展[J].天津中医药大学学报,2007,26(1):54-56
30陆网坤.绿茶提取物对人肺癌细胞影响的初步探讨[J]中国现代药物应用,2008,2(3):38-39
2周曾同.提高口腔黏膜癌前病变的诊疗质量[J].中华口腔医学杂志,2005,40(2):89-91
3刘伟,周曾同.口腔白斑的中医药治疗现状及研究进展[J].临床口腔医学杂志,2009,25(1):53-55
4 The Ministry of Science and Technology of the People's Republic of China. Guidance suggestion of caring laboratory animals.2006-09-30
5周曾同,娄佳宁.绞股蓝总甙对金地鼠颊囊白斑癌变过程中端粒酶活性化学预防作用研究[J].临床口腔医学杂志,2008,24(4):234-237
6李萍,葛化冰,孙正.酪氨酸激酶抑制剂GW2974对口腔鳞状细胞癌生长抑制作用的实验研究[J].中华口腔医学杂志,2007,42(7):432-435
7许彦枝,辛晓红,刘亚娴,等.藓化饮对口腔扁平苔藓患者激活调节因子表达的影响[J].山东中医,杂志,2009,28(1):13-14
8许良中,杨文涛.免疫组织化学反应结果的判定标准[J].中国癌症杂志,1996,6(4):229-231
9吴明利,王士杰,徐志彬,等.家兔颊黏膜鳞状上皮癌前病变模型制备及癌前病变逆转研究[J].实用癌症杂志,2005,20(1):5-7
10刘栋才,杨竹林,杨乐平.胆囊良恶性病变组织EZH2和PTEN表达及意义[J].中南大学学报,2008,33(7):618-622
11 Gil J, Bernard D, Peters G. Role of polycomb group proteins in stem cell self-renewal and cancer [J].DNA Cell Biol, 2005, 24(2):117- 125
12苗芳,李劲松.EZH2和Ki-67在胃癌中的表达及其临床意义[J].泰山医学院学报,2008,29(2):87-89
13黄优,王林,马俊青,等.大鼠生长发育过程中颅底软骨联合细胞增殖和凋亡的研究[J].口腔医学,2008,28(2):57-60
14李晓光,于肖鹏,赵海兰,等.PCNA在口腔鳞癌中的表达及其临床意义[J].泰山医学院学报,2005,26(1) :20-22
15 Bracken AP , Pasini D , Capra M, et al. EZH2 is downstream of the pRB- E2F pathway , essential for proliferation and amplified in cancer[J ] . EMBO , 2003 , 22 (20) : 5 323- 5 335
16王文斌,祁佩时.电子显微技术研究生物膜特性[J].工业安全与环保,2008,34(7):15-16
17范刚启,宋祥龙,王辉,等.活血化瘀治疗癌及癌前病变效应的两重性与血管生成的关系[J].中国中西医结合杂志,2003,23(8):624-626
18魏道智,宁书菊,林文雄.佩兰的研究进展[J].时珍国医国药,2007,18(7):1782-1783
19高贵峰,杜作鹏.浅谈蝉蜕在临床上的多功能应用[J].中华实用中西医结合杂志,2008,21(8):650-651
20许继文,付春梅.丹参的药理作用研究进展[J].医学综述,2006,12(23):1467-1468
21杨媛媛,周刚,马晓康,等.赤芍的研究进展[J].医药导报,2008,27(1):67-69
22许红兰.苦参的药理研究进展[J].现代医药卫生,2008,24(20):3066-3067
23郑利光,翟所迪.甘草锌膜对实验性口腔溃疡的治疗作用及黏膜刺激性研究[J].中国新药杂志,2006,15(8) :601-603
24刘庆林.赤芍的临床配伍应用浅析[J].中国新药杂志,2009,24(1):131-132
25王晓菲,金鸣.红花抗炎作用机制研究进展[J].中国新药杂志,2007,36(1) :51-53
26徐萍利,雷正荣.中医药防治胃癌前病变的研究进展[J].中外健康文摘,2008,5(1) : 1- 4
1周曾同.提高口腔黏膜癌前病变的诊疗质量[J].中华口腔医学杂志,2005,40(2):89-91
2刘伟,周曾同.口腔白斑的中医药治疗现状及研究进展[J].临床口腔医学杂志,2009,25(1):53-55
3 Bokor-Bratic M.Prevalence of oral leukoplakia. Med Pregl[J]. 2003, 56 (11-12): 552-555
4姜倩,祝磊,高黎.口腔白斑发病因素的初步探讨[J].河南医学研究,2006,15(4):363-365
5高岩. WHO口腔黏膜癌前病变组织学新分类及其进展[J].中国实用口腔科杂志,2008,1(12):705-708
6周曾同,钟文静,张水龙.微波对白斑上皮异常增生12项病理特征出现率的影响[J].临床口腔医学杂志,1998,14(3):163-165
7周曾同,张水龙,王英,等.金地鼠颊囊上皮异常增生12项病理特征的光镜分析[J].上海口腔医学,1997,6(1):32-35
8葛姝云,周曾同.口腔癌前病变生物标志物的研究进展[J].中华口腔医学杂志,2005,40(2):166-168
9孟宪瑞,刘进忠.口腔癌的早期诊断[J].国际口腔医学杂志,2008,35(3):329-331
10孙善珍.口腔癌前病变的诊断及其恶变潜能的监测[J].山东医药,1997,37(10):33
11周曾同,张水龙.口腔黏膜白斑研究概况[J].中华口腔医学杂志,1999,34(6):382-384
12庞劲凡.口腔白斑的药物治疗[J].武警医学,2008,19(4):349-352
13黄霞萍.益气活血、通络解毒法治疗口腔扁平苔藓30例[J].中医研究,2008,21(12):28-30
14王贵林.灯盏细辛注射液对不稳定心绞痛患者血流变学的影响[J].中国现代药物应用,2009,3(2):121-122
15娄佳宁,周曾同.绞股蓝总甙对金地鼠颊囊白斑癌变过程中端粒酶活性化学预防作用研究[J].临床口腔医学杂志,2008,24(4):234-236
16张为新,胡慧芳,李伟国,等.7种中药对实验性口腔癌阻断作用的研究[J].上海口腔医学,2004,13:34-37
17许彦枝,辛晓红,刘亚娴,等.藓化饮对口腔扁平苔藓患者激活调节因子表达的影响[J].山东中医杂志,2009,28(1):13-14
18张玉禄,李军祥,朱陵群.三种活血法对大鼠萎缩性胃炎癌前病变早期细胞凋亡的影响[J].中国中西医结合杂志,2008,28(5):448-450
19郑利光,翟所迪.甘草锌膜对实验性口腔溃疡的治疗作用及黏膜刺激性研究[J].中国新药杂志,2006,15(8):601-603
20陈谦明,李秉琦.口腔白斑病诊断、分级与治疗中的新观点[J].华西口腔医学杂志,2004,22(2):142-144
21林梅.口腔癌前病变的临床检查和风险评估[J].中华口腔医学研究杂志(电子版),2008,2(5):1-3
22周曾同.口腔白斑、红斑和黑斑的诊断与治疗[J].中华口腔医学杂志, 2006,41(8):502-505
23常云龙,左德崴.微波治疗舌下腺囊肿92例临床观察[J].中国医学创新,2008,5(35):52
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