阻断交感神经对小鼠小肠黏膜免疫相关细胞的影响

目的研究交感神经对肠道黏膜屏障的调节作用。方法采用常规组织学和免疫组织化学技术,观察经腹腔注射交感神经损毁剂6-羟多巴胺后小鼠小肠上皮内淋巴细胞、杯状细胞、IgA+浆细胞、肥大细胞的数量分布变化。结果注射6-羟多巴胺组小鼠与对照组小鼠相比,小肠各肠段的上皮内淋巴细胞减少了5.6%～33.8%,杯状细胞减少了10.6%～18.5%,两组指标在十二指肠之间和回肠之间差异极显著(P<0.01);肥大细胞增加了15.0%～57.1%,在空肠之间差异极显著(P<0.01);IgA+浆细胞增加了13.9%～61.4%,在十二指肠之间差异显著(P<0.05)。结论交感神经对调节小鼠肠道的免疫学屏障有着重要作用。     

大鼠视网膜发育过程中细胞增殖与凋亡的形态学

目的:观察视网膜发育过程中增殖细胞核抗原(PCNA)的表达变化及细胞凋亡状态。方法:14~20 d胎鼠(E14-20 d)、生后0~15 d幼鼠(P0-15)及成年鼠(P36),取出眼球组织,免疫组织化学显色及原位末端标记(TUNEL)观察PCNA的表达和细胞凋亡变化。结果:发育早期E14~P7,可观察到有较强PCNA阳性表达,P9时PCNA阳性细胞数目明显减少,P15开始未见明显PCNA阳性表达。E18可观察到凋亡细胞,随着发育的进行,凋亡细胞逐渐增多,P7时凋亡细胞数目最多,之后又逐渐减少,成熟的视网膜组织内未见明显凋亡细胞。结论:在视网膜神经细胞发育过程中,细胞的增殖和凋亡呈现了有序的动态变化,正是这种有序的增殖和凋亡的平衡才使视网膜最终发育成了正常的组织结构及功能。     

脑星形细胞瘤中bFGF及其受体对血管新生 和肿瘤细胞增殖的影响

目的：为探讨碱性纤维母细胞生长因子（ｂＦＧＦ）及其受体（ＦＧＦＲ－２）对人星形细胞瘤的血管新生及细胞增殖的作用。方法：彩和免疫组织化学方法,结果：发现ｂＦＧＦ及其受体,增殖细胞核抗原（ＰＣＮＡ）在瘤细胞及血管内皮细胞中均有表达,高级别星形细胞瘤的表达阳性率高于低级别者,ｂＦＧＦ在瘤细胞中的表达阳性率分别为与ＦＧＦＲ－２及ＰＣＮＡ的表达阳性率呈正相关关系（Ｐ〈０．０１,Ｐ〈０．０５）,ｂＦＧＦ及其受     

脑星形细胞瘤中bFGF及其受体对血管新生和肿瘤细胞增殖的影响

目的：为探讨碱性纤维母细胞生长因子（ｂＦＧＦ）及其受体（ＦＧＦＲ－２）对人星形细胞瘤的血管新生及细胞增殖的作用。方法：彩和免疫组织化学方法,结果：发现ｂＦＧＦ及其受体,增殖细胞核抗原（ＰＣＮＡ）在瘤细胞及血管内皮细胞中均有表达,高级别星形细胞瘤的表达阳性率高于低级别者,ｂＦＧＦ在瘤细胞中的表达阳性率分别为与ＦＧＦＲ－２及ＰＣＮＡ的表达阳性率呈正相关关系（Ｐ〈０．０１,Ｐ〈０．０５）,ｂＦＧＦ及其受     

不同固定剂对检测增殖细胞核抗原表达的影响

不同固定剂对检测增殖细胞核抗原表达的影响牛兆山邹伟项锋刚郭成浩增殖细胞核抗原（ｐｒｏｌｉｆｅｒａｔｉｎｇｃｅｌｎｕｃｌｅａｒａｎｔｉ－ｇｅｎ，ＰＣＮＡ）作为评价细胞增殖指数的指标之一，已广泛地应用于肿瘤病理学研究中。但许多证据表明，ＰＣ－ＮＡ的免疫反...     

髓母细胞瘤增殖活性的研究及其与预后的关系

增殖细胞核抗原（ＰＣＮＡ）为ＤＮＡ合成中聚合酶δ的辅助蛋白。ＰＣＮＡ的水平直接与ＤＮＡ的合成和细胞的增殖密切相关，并在其中发挥重要作用〔１〕。因此，ＰＣＮＡ的表达反映了细胞增殖活性〔２〕。ＰＣＮＡ表达的水平与肿瘤的预后和分化程度等方面也有一定的关系〔...     

阻断交感神经对青年和老龄小鼠小肠T淋巴细胞活性的影响

目的探究交感神经在青年和老龄小鼠中对小肠T淋巴细胞的免疫调控机制。方法选用健康的3月龄和18月龄的雄性昆明小鼠,腹腔注射六羟多巴胺,通过石蜡切片和HE染色观察上皮内淋巴细胞的数量,MTT法测定小肠T淋巴细胞的增殖,ELISA试剂盒测定小肠T淋巴细胞分泌IL-2、IL-4水平。结果阻断交感神经后,2个年龄组上皮内淋巴细胞数量均减少,老龄组尤为突出。青年小鼠T淋巴细胞增殖在处理后第5、10天出现下降,老龄小鼠在第3天至第15天均显著下降,其中第5天下降39.4%(P<0.01)。青年小鼠损毁交感神经后主要引起淋巴细胞分泌IL-4含量降低,而老龄小鼠主要引起IL-2降低。结论损毁交感神经可导致青年和老龄小鼠小肠T淋巴细胞活性下降,老龄组下降幅度大,恢复周期长。     

靶向PCNA基因的小干扰RNA对鼻咽癌CNE2细胞周期的影响

目的：研究小干扰RNA抑制PCNA基因表达后对鼻咽癌CNE2细胞株增殖及细胞周期的影响。方法:利用体外合成法合成针对PCNA基因的小干扰RNA(siRNA)序列,应用脂质体转染鼻咽癌CNE-2细胞,实时定量PCR(real-timePCR)和免疫组织化学法观察siRNA干扰后细胞中PCNAmRNA水平和PCNA蛋白表达,倒置相差显微镜观察转染前后CNE2细胞的生长变化,流式细胞仪检测siRNA干扰后的细胞周期。结果:在siRNA转染CNE2细胞后,细胞增殖受到抑制,蛋白表达不同程度降低,对PCNAmRNA水平的抑制达98.5%,细胞周期在G0/G1停滞。结论:siRNA可在鼻咽癌中有效发挥RNA干扰效应,抑制PCNAmRNA及蛋白表达,降低PCNA活性,抑制鼻咽癌细胞增殖,为鼻咽癌基因治疗提供了新的方法。     

HBx及其羧基末端40个氨基酸对SMMC-7721肝细胞增殖的影响

目的研究HBx羧基末端40个氨基酸对肝癌细胞增殖的影响及作用机制。方法用HBx及去除羧基末端40个氨基酸HBx3′40转染含野生型(wt)p53的SMMC7721细胞,经稳定筛选后,通过细胞生长曲线绘制以及平板克隆形成实验来观察HBx对细胞增殖的影响,并通过裸鼠体内的成瘤实验观察各移植瘤的大小及重量、增殖细胞核抗原(PCNA)表达的变化。结果细胞生长曲线表明HBx及HBx3′40的细胞生长速度明显较空载体增快;平板克隆形成试验表明克隆形成率,HBx组(48.7±8.1)%及HBx3′40组(82.8±6.0)%较对照组(26.9±3.5)%明显增多(P<0.05),pcDNA3HBx3′40组又较pcDNA3HBx组增多(P<0.05)。裸鼠成瘤实验pcDNA3HBx3′40组肿瘤的大小及重量(1.476±0.232、0.987±0.279)%及pcDNA3HBx组(0.412±0.212、0.395±0.159)%较对照组(0.051±0.024、0.033±0.004)%明显增大(P<0.05),肿瘤细胞的PCNA表达率pcDNA3HBx3′40组(69.25±3.77)%及pcDNA3HBx组(59.00±2.58)%也较对照组(37.67±2.52)%明显增强(P<0.05),且以HBx3′40组的表现更为明显(P<0.05)。结论HBx的羧基末端40个氨基酸可刺激细胞增殖,其突变可能与肝癌细胞发生过程中促进癌细胞的恶性转化有关。     

桃核承气汤对Bcl-2和增殖细胞核抗原蛋白表达的影响

目的探讨桃核承气汤(THCQT)对S180荷瘤小鼠环磷酰胺化疗后的Bcl-2和增殖细胞核抗原(PCNA)蛋白表达的影响.方法建立小鼠S180肿瘤模型,采用免疫组织化学方法测定S180小鼠Bcl-2和PCNA蛋白表达的情况.结果 Bcl-2蛋白的表达THCQT组与模型组比较t=5.29,P＜0.01,差异有统计学意义,T...     

小檗碱和育亨宾对LPS诱导的小鼠肠道损伤和肠上皮细胞增殖抑制的影响

目的:观察小檗碱(Ber)和育亨宾(Y)对LPS诱导的小鼠肠道损伤和肠上皮细胞增殖抑制的影响。方法:雄性BALB/c小鼠随机分为对照组(control)、脂多糖组(LPS)、小檗碱组(Ber+LPS)、小檗碱与育亨宾合剂组(Ber+Y+LPS)、育亨宾组(Y+LPS)、小檗碱组(Ber)、小檗碱和育亨宾合剂组(Ber+Y)及育亨宾组(Y)。分别予以双蒸水、Ber(50mg/kg)、Ber(50mg/kg)+Y(2mg/kg)、Y(2mg/kg)灌胃,1time/d,连续3d,于实验第3d灌胃后1h,腹腔注射生理盐水或LPS(18mg/kg,0.2mL/10g)。观察各组小鼠肠组织形态学、肠黏膜重量和绒毛高度的改变;酶联免疫吸附实验(ELISA)测定肠组织二胺氧化酶(DAO)含量;免疫组化染色方法分析各组肠组织增殖细胞核抗原(PCNA)的表达。结果:LPS组小鼠肠腔显示炎性渗出、出血;肠损伤评分明显高于对照组;LPS组肠黏膜重量、肠绒毛高度、肠组织DAO含量、肠上皮细胞PCNA表达明显低于对照组。与LPS组比较,Ber组、Ber+Y组上述改变明显减轻,但两组间未见明显差异;育亨宾对LPS引起的上述指标变化无明显抑制作用。结论:Ber通过非α2肾上腺素能受体依赖的途径减轻LPS引起的肠上皮细胞增殖抑制和肠道损伤。     

甘油致BALB/C小鼠急性肾损伤后肾脏观察

目的 观察甘油致小鼠急性肾脏损伤后肾脏修复的情况.方法 肌肉注射甘油致小鼠发生急性肾脏损伤后出取肾脏组织,于损伤后的不同时间点进行肾脏组织形态学检测;同时,应用免疫组织化学技术检测核增殖抗原在肾脏组织的表达情况,分析肾脏损伤后的修复情况.结果 肌肉注射甘油3 d后小鼠开始出现急性肾脏损害的表现.主要表现为肾脏肿胀,肾脏...     

胶质瘤增殖活性和肿瘤基因蛋白表达及其意义

目的探讨胶质瘤增殖活性和肿瘤基因蛋白表达与分化和预后间的关系。方法应用免疫组化和图像分析技术对１２４例脑胶质瘤增殖细胞核抗原（ＰＣＮＡ）和几种肿瘤基因蛋白进行定性、定量研究。结果ＰＣＮＡ反应强度与级别和预后显著相关;ｃ－ｅｒｂＢ－２蛋白表达在分化程度高的胶质瘤强于分化程度低者,存活５年以上的病例阳性强于存活５年以下者。ＥＧＦ（４０．０％）、ＥＧＦＲ（９１．４％）和ｐ２１ｒａｓ（５３．３％）与分级和预后无显著关系。３种ｐ５３蛋白抗体阳性率均以Ⅱ～Ⅳ级胶质瘤为高;ｐ５３与ＰＣＮＡ反应强度平行。结论ｐ２１ｒａｓ、ｃ－ｅｒｂＢ－２、ＥＧＦ和ＥＧＦＲ改变在胶质瘤发生、发展过程中可能是早期事件,恶变后不再随级别增加而加重,而ｐ５３基因突变涉及胶质瘤发展各个阶段;ＰＣＮＡ能较好地反映胶质瘤的恶性程度。     

乳腺叶状囊肉瘤的增殖细胞核抗原的表达及其意义

目的研究乳腺叶状囊肉瘤增殖细胞核抗原（ＰＣＮＡ）的表达及其临床病理意义。方法采用免疫组化ＳＰ法对１００例乳腺叶状囊肉瘤及３９例乳腺腺纤维瘤进行检测。结果叶状囊肉瘤中间质性瘤细胞呈ＰＣＮＡ阳性着色者８６例（８６％），不同组织学级别者ＰＣＮＡ指数之间有显著差异（Ｆ＝８５．３３，Ｐ＜０．０１），分化越差，ＰＣＮＡ平均指数越高，ＰＣＮＡ指数与组织学分级密切相关（ｒ′ｓ＝０．７７）。Ⅰ级叶状囊肉瘤与乳腺腺纤维瘤间质性瘤细胞丰富组的ＰＣＮＡ指数也有显著差异（ｔ＝３．４２，Ｐ＜０．０１）。ＰＣＮＡ指数与核分裂相计数间相关关系并不密切（ｒ＝０．３９）。结论用ＰＣＮＡ来反映乳腺叶状囊肉瘤增殖活性，辅助进行组织学分级，鉴别良恶交界性病变及估测预后，都具有一定的实用价值。     

Proliferation kinetics of streptozotocin-induced renal tumours in mice

Renal tumours were induced in female mice 132 days after intravenous administration of streptozotocin. The tumours exhibited papillary and/or solid architecture with papillary tumours showing no histological evidence of malignancy. Malignant behaviour, manifest as infiltration of adjacent renal tissue and lymphatic infiltration, was noted for tumours with solid architecture. Intermediate architectural forms exhibiting dual papillary and solid architecture were identified. Proliferation kinetics were evaluated by enumeration of silver-staining nucleolar organizer regions and proliferating cell nuclear antigen expression. The results showed a stepwise progress in cell proliferation between histologically normal renal tubule epithelium [untreated animals, mean AgNOR score (MAS) 2.32, mean PCNA index (MPI) 0.53%; treated animals, MAS, 2.44; MPI 0.99%], dysplastic tubule epithelium (MAS, 4.15; MPI 1.65%), papillary tumours (MAS, 5.90; MPI 3.89%) and solid tumours (MAS, 6.94; MPI, 6.80%). Solid tumours as a group were significantly larger than papillary tumours and were associated with a significantly longer mean post-injection survival interval. The findings suggest that at least some solid tumours evolve from tumours exhibiting papillary architecture.     

养血方与养血解毒方对咪喹莫特诱导的银屑病样小鼠的作用

目的:探讨并比较养血方与养血解毒方对银屑病样小鼠模型皮损的干预作用。方法:将50只BALB/c小鼠随机分为空白对照组、模型组、甲氨蝶呤组、养血方组和养血解毒方组,每组10只,采用背部涂抹咪喹莫特的方式诱导银屑病样模型。动态观察皮损变化并进行银屑病皮损面积和严重程度指数(PASI)评分;通过皮肤水/油测试笔检测小鼠背部皮肤水/油含量;HE染色观察皮损病理改变并检测表皮厚度;免疫组化法检测皮损表皮中增殖细胞核抗原(PCNA)和真皮中CD3+T淋巴细胞表达;real-time PCR法检测皮损中白细胞介素(IL)-17、IL-23和IL-1β的mRNA相对含量。结果:养血方组、养血解毒方组和甲氨蝶呤组背部皮损表现均优于模型组,PASI评分和表皮厚度均低于模型组,皮肤水分/油分含量高于模型组(P0.05),养血解毒方组背部皮损表现和皮肤水分/油分含量优于养血方组,PASI评分和表皮厚度低于养血方组;养血方组、养血解毒方组和甲氨蝶呤组皮损表皮PCNA表达与真皮CD3+T细胞表达均低于模型组(P0.05),养血解毒方组表皮PCNA表达与真皮CD3+T细胞表达均低于养血方组;real-time PCR显示养血方组与养血解毒方组IL-17、IL-23和IL-1β的mRNA相对表达量低于模型组(P0.05),养血解毒方组IL-1β的mRNA相对表达低于养血方组。养血解毒方组与甲氨蝶呤组比较,皮损表现无明显差异,皮肤水/油分含量和对炎症因子的抑制作用优于甲氨蝶呤组。结论:养血方与养血解毒方可通过降低T细胞活化相关因子含量,减轻免疫反应,改善咪喹莫特诱导的银屑病样小鼠皮损;同时养血解毒方效果优于养血方。     

Proliferating cell nuclear antigen in malignant and pre-malignant lesions of epithelial origin in the oral cavity and the skin: an immunohistochemical study

Summary Proliferating cell nuclear antigen (PCNA) is a nuclear protein synthesized in the late G₁ and S phase of the cell cycle and immunohistochemical detection of the protein represents a useful marker for the proliferating fraction of cells in tissue specimens. A series of malignant and pre-malignant lesions of the oral cavity and skin were evaluated by the streptavidin biotin immunoperoxidase method for detection of this protein. Monoclonal anti-PCNA antibody (PC 10) labelled proliferating cells in all cases with varying intensity of nuclear staining. In squamous cell carcinoma (n=48), PCNA positivity correlated with the differentiation and atypia of the tumour cells; however, in poorly differentiated tumours, the relationship between PCNA expression and proliferation was lost. Basal cell carcinoma showed an increased growth fraction in tiny epithelial nests (mean 43.8, SD 6.0,n=20) than in neoplastic basal cells (mean 30.1, SD 6.9,n=8). The growth fractions were significantly higher in the pre-malignant lesions (leukoplakia, mean 22.3, SD 7.7,n=14; Bowen's disease, mean 45.2, SD 11.7,n=12; senile keratosis, mean 41.2, SD 7.0,n=12) than in the normal mucosa (mean 9.8, SD 4.9,n=10), suggesting that cellular growth fractions correlate with the degree of dysplasia in pre-malignant lesions.     

结直肠癌、结直肠腺瘤微卫星不稳定性检测及其临床意义

目的 探讨微卫星不稳定性（ＭＳＩ）在结直肠癌发生中的意义及其与结直肠癌细胞增殖活性和预后的关系,方法 采用聚合酶链反应－简单序列长度多态性（ＰＣＲ－ＳＳＬＰ）及免疫组化ＳＰ方法,对５６例结直肠癌,９例腺瘤及６例腺瘤癌变的ＭＳＩ和增殖细胞核抗原（ＰＣＮＡ）表达进行了检测。结果 显示结直肠癌ＭＳＩ总阳性率为４４．６４％（２５／５６例）,共中遗传性非息肉病性结直肠癌３／４例阳性,散发性结直肠癌２２／５２     

Neonatal sympathectomy by 6-hydroxydopamine: cardiovascular responses in the paralyzed rat

Rats who had been neonatally sympathectomized by peripheral injections of 6-hydroxydopamine or vehicle were paralyzed by succinylcholine and received either atropine or saline injection. Heart rate and blood pressure responses to foot shocks and to a conditioned stimulus (CS) for shock were continuously recorded. Sympathectomized rats showed lower basal systolic blood pressure, less second to second blood pressure variability and attenuated pressor responses to the shock and the CS. Basal heart rate and the heart rate response to shock were were unaffected by sympathectomy, but cardiac rate deceleration to the CS was ablished. Atropine increased basal heart rate, decreased heart rate variability and abolished the cardiac deceleration to the CS. These data were interpreted as suggesting that conditioned cardiac deceleration in the rat is a vagal mediated response compensatory to prior blood pressure increases. Sympathectomized rats also showed higher core temperatures under paralysis.     

Immunoreactivity of proliferating cell nuclear antigen in salivary gland tumours: An assessment of growth potential

Immunoreactivity of proliferating cell nuclear antigen (PCNA) was assessed to evaluate growth potential in surgically resected tissue specimens from 70 cases of benign and malignant salivary gland tumours. Three stage streptavidin-biotin immunoperoxidase immunostaining using monoclonal antibody to PCNA showed a heterogeneity of PCNA index and distribution. In normal salivary gland specimens, PCNA was demonstrated in the nuclei of few ductal and acinar cells. In pleomorphic adenoma a multiple nodular growth pattern was observed with positive immunoreactivity restricted to the nuclei of tubulo-ductal structures. Warthin's tumour had positive nuclei in the outer cuboidal cells of epithelial component and germinal centres of lymphoid tissue. Myoepithelioma and acinic cell carcinoma showed slightly differing values and a statistically significant difference in the value of the index was observed in tumour cell aggregates of the cribiform type of adenoid cystic carcinoma and the solid undifferentiated type and between low/intermediate and high-grade mucoepidermoid tumours. PCNA is a useful marker of tumour cell proliferation; the index correlates with the grade of malignancy in salivary gland tumours.