制作角膜缘干细胞完全缺乏动物模型的新方法

目的提出一种新的角膜缘干细胞完全缺乏模型制作方法,为干细胞缺乏疾病的治疗性研究提供可靠的模型。方法利用9mm环钻划界,板层切除角膜组织,通过临床评分、印迹细胞学、组织病理及免疫组织化学验证模型的成功。结果板层切除术后30~45d,模型眼角膜混浊、上皮缺损、大量新生血管长入,印迹细胞学可见PAS( )的杯状细胞,组织病理学检查上皮符合结膜细胞表型,免疫荧光染色可见结膜杯状细胞特异性标志MUC5AC阳性表达。结论环钻划界加板层切除法为一种可靠、有效的角膜缘干细胞完全缺乏模型制作方法。     

宫颈环形电切术对阴道镜活检诊断为宫颈上皮内瘤变的再评估

目的通过宫颈环形电切术(LEEP)对阴道镜活检诊断为宫颈上皮内瘤变(C IN)的准确性和全面性进行评估。方法对118例阴道镜活检诊断为C IN的患者行LEEP,对比术前术后的病理结果。结果32例阴道镜活检病理为C INⅠ级病例中有11例与LEEP术病理相符,6例LEEP术后病理级别上升,15例LEEP术后病理级别下降;26例阴道镜活检病理C INⅡ级病例中有17例LEEP术后病理示相符,5例LEEP术后病理级别上升,4例LEEP术后病理级别下降;60例阴道镜活检病理为C INⅢ级病例中有44例LEEP术后病理示相符,11例LEEP术后病理级别上升,5例术后病理级别下降。结论LEEP术是明确诊断和治疗C IN的有效而理想的方法,阴道镜活检有一定局限性。     

双光气致急性损伤对兔肺泡Ⅱ型细胞形态学的影响

目的：通过对日本大耳兔双光气肺损伤后肺泡Ⅱ型细胞内磷脂和形态学变化的观察，探讨双光气对肺Ⅱ型细胞的损伤及其机制．方法：将纯种日本大耳白兔35只随机分为对照组(n＝10)、染毒即刻组(n＝5)、染毒2h组(n＝5)染毒4h组(n＝5)、染毒8h组(n＝5)、染毒12h组(n＝5)；采用浓度一时间法动态式染毒，染毒后按不同时间点对实验动物进行颈总动脉插管放血处死，取肺并用肺灌洗液对支气管肺泡进行灌洗，然后取肺组织作冰冻切片，进行电镜观察，并测定细胞内磷脂含量．结果：肺Ⅱ型细胞内磷脂含量在染毒即刻开始时较正常组含量下降(P＜0．01)；在染毒2h时降到最低，之后上升(P＜0．05)，而在4h后又下降(P＜0．05)．染毒终止后，损伤逐渐加重，染毒后2h磷脂因消耗而降低．染毒后4h，由于代偿功能启动，磷脂短暂升高，出现致伤性代偿期．从染毒2h后开始，细胞损伤加重，磷脂合成进入失代偿期，合成逐渐减少．到染毒后8h，肺泡Ⅱ型细胞内细胞处于致伤性和致死性细胞改变的临界点，细胞出现致死性改变，磷脂合成、分泌停止，细胞出现死亡．结论：兔在双光气染毒12h内，肺泡表面活性物质磷脂的改变有差异，随细胞损伤程度发生改变，出现明显中毒分期：即致伤性改变期、致伤性代偿期、致伤性失代偿期、致死性改变期．     

Aspiration biopsy cytology of mammary myoepithelioma

A case of ultrastructurally confirmed myoepithelioma of the breast studied by fine-needle aspiration biopsy is reported. Aspirated tumor cells were spindle-shaped with well-defined cytoplasm and oval nuclei showing no remarkable nuclear pleomorphism. They were seen mainly in irregular bundles with strong cellular cohesiveness. Their cytoplasm contained S-100 protein demonstrated by the peroxidase antiperoxidase staining technique.     

花生四烯酸和二十二碳六烯酸对海马神经细胞生长发育的影响

【目的】观察不同浓度花生四烯酸 (AA)和 (或 )二十二碳六烯酸 (DHA)对体外培养海马神经细胞生长发育的影响。【方法】无血清培养液中分别加入不同剂量的AA和 (或 )DHA ,采用噻唑蓝比色试验 (MTT法 )反映各组海马神经细胞存活状况 ,神经元特异性烯醇化酶 (NSE)染色鉴定神经细胞 ,图像分析技术测量神经细胞的胞体大小和突起长度。【结果】培养液中分别添加AA为 4～ 8μmol/L、DHA为 2～ 8μmol/L时 ,神经细胞活力高于对照组 ;当培养液中AA和DHA总浓度为 4μmol/L、比例为 2∶1或 4∶1时 ,神经细胞的活力、胞体面积、最大长径、最大短径和平均突起长度均显著高于单一添加 4μmol/LAA、4μmol/LDHA和对照组。【结论】AA、DHA均具有促进体外培养海马神经细胞生长发育的作用 ;与单独添加AA、DHA相比 ,AA和DHA共同作用的促海马神经细胞生长发育作用更好。     

Interactions between peripheral blood CD8 T lymphocytes and intestinal epithelial cells (iEC)

Intestinal intraepithelial lymphocytes (iIEL) are primarily CD8 cells and most of them have a CD28^? phenotype, the phenotype of effector cytotoxic T cells. We asked whether the predominance of CD8⁺ CD28^? T cells in the gut may result from peripheral blood T cells preferentially migrating to the iIEL compartment and adhering to iEC. Compared with CD4 cells, adhesion of resting CD8⁺ T cells to iEC cell lines was significantly higher. Adhesion could be blocked with a MoAb to gp180, a molecule expressed on iEC which is known to interact with CD8/lck. No significant difference in the level of adhesion was observed between CD8⁺ CD28⁺ and CD8⁺ CD28^? T cells. Thus CD8 cells may preferentially migrate to the iIEL compartment, but loss of CD28 expression could occur in situ after migration. Consistent with this hypothesis, the CD8⁺ CD28^? cells became enriched after co-culturing T cells with iEC cell lines and primary iEC. Induction of the CD8⁺ CD28^? phenotype in cord blood and adult T cells was observed in co-cultures with iEC and also with mitogens and superantigens. In the latter case, CD28 down-modulation was seen specifically in the Vβ subset targeted by the superantigen, indicating that loss of CD28 expression is a direct result of T cell receptor (TCR)-mediated stimulation. The combined results suggest that CD8⁺ CD28^? T cells are antigen experienced T cells, and that they may have a survival advantage in the presence of gut epithelial cells in vitro. This may contribute to the predominance of CD8⁺ CD28^? T cells in the iIEL compartment.     

Granular cell tumors of the breast: A report of two cases

Granular cell tumors (GCT) are rare neoplasms, and only 173 cases of benign GCT of the breast have been documented. We report herein the cases of two patients with this tumor and discuss the methods of diagnosis and treatment. The first patient was a 60-year-old woman who presented with a firm ill-defined mass in her left breast. Mammography showed a dense shadow with spicula and skin thickness, and ultrasonography revealed a hypoechoic mass with an irregular border. Radical mastectomy was performed under the wrong preoperative diagnosis of breast cancer. The second patient was a 31-year-old woman who presented with an elastic-hard mass in her left breast. Mammography showed a well-demarcated dense mass, and ultrasonography revealed a well-defined hypoechoic mass with a large depth-width ratio. Fine-needle aspiration cytology (FNAC) showed a large number of histiocytic cells with abundant granular cytoplasm. An excisional biopsy was performed, and histological examination confirmed a diagnosis of GCT. GCT is benign, but often misdiagnosed as breast cancer both clinically and radiologically. Therefore, histological examination is essential for making a correct diagnosis, while FNAC is also useful. Local resection is still the treatment of choice, and surgeons should do their utmost to avoid performing needless radical mastectomy.     

兔关节软骨细胞的分离、培养和形态学特征

[目的]探讨兔关节软骨细胞的分离、培养方法,观察单层高浓度培养时细胞表型表达情况.[方法]无菌条件下,从 2周龄新西兰白兔的颞颌关节及四肢关节髁突面削取软骨片,采用机械-酶消化法分离软骨细胞,经台盼蓝拒染计数,将细胞按 1× 106个 /孔接种于 6孔培养板,传代培养,描绘生长曲线.利用相差显微镜及透射电镜观察细胞形态.应用甲苯胺蓝及Ⅱ型胶原免疫组化对细胞进行鉴定.[结果]每克软骨能获取 1 5× 106个软骨细胞,活性率为 95%.培养 2～ 3 d,细胞贴壁、变形,呈多角形; 8 d左右,细胞融合成层.透射电镜观察显示细胞核圆形,有丰富的粗面内质网、高尔基体及分泌的基质成分.甲苯胺蓝及Ⅱ型胶原染色阳性.细胞传至 5代后,出现"成纤维细胞样".[结论]本研究建立了简单易行的软骨细胞分离、培养方法;初代、第 2代细胞生长良好,适合于实验研究;软骨细胞 5代培养后,细胞表型发生改变.