Flow Cytometric Analysis of Lymphocyte Subsets of Mice Maintained on an Ethanol-Containing Liquid Diet

Alcoholic patients often have impaired immune function, yet little is known about the precise mechanism(s) of this impairment. We have previously shown that ethanol consumption by mice alters copolymer-specific humoral and cellular immune responses. In this study, we asked whether alcohol consumption by mice would phenotypically alter lymphocyte populations. Female C57BL/6 mice were fed a nutritionally complete liquid diet containing 35% ethanol-derived calories for up to 8 days. As controls, mice either were fed a liquid control diet that isocalorically substitutes sucrose for ethanol or remained on a standard solid diet and water ad libitum. Although mice fed ethanol-containing liquid or pair-fed control liquid diets have decreased numbers of spleen cells compared with solid diet controls, only the ethanol-containing diet allowed normally nonresponder C57BL/6 spleen cells to make antibody responses to the poly(Glu⁵⁰Tyr⁵⁰) synthetic copolymer antigen. Flow cytometric analysis of splenic lymphocyte populations of mice on the ethanol-containing diet shows an increase in the relative proportion of T-lymphocytes as compared with mice on either solid or liquid control diets. No such change is seen for either B-cell or natural killer cell populations in these same mice. Both liquid control and liquid ethanol diets caused a slight decrease in the CD4:CD8 ratios of splenic T-lymphocytes. We see the relative percentage of T-cells bearing the αβ-cell receptor (TcR) increases in the spleens of liquid ethanol diet mice; a smaller increase TcRαβ usage is seen in the spleens of liquid control mice, compared with solid diet mice. Flow cytometric analysis shows that little, if any, difference exists in TcRγδ expression between the liquid ethanol and either the liquid control or solid diet groups. Preliminary analysis of TcRαβ subsets suggest that ethanol increases the percentage of T-cells expressing Vβ5 and Vβ8, and decreases the percentage of Vβ11 expressing cells. These findings suggest that, in addition to modifying the immune response, ethanol alters the phenotypic expression of lymphocyte subsets.     

应用流式细胞仪对肺癌细胞增殖活力的分析

本文应用流式细胞仪对35例肺癌、10例良性病变肺组织及10例正常肺组织的细胞增殖活力(S％)进行了分析,结果显示肺癌组织细胞增殖活力非常显著高于良性病变组,良性病变组细胞增殖活力亦非常显著高于正常组,提示检测细胞增殖活力对肺癌的诊断、筛选有一定意义。     

子宫肌瘤与细胞凋亡的研究

目的:探讨子宫平滑肌瘤的发生与细胞凋亡的关系.方法:采用凝胶电泳技术对子宫平滑肌瘤患者的肌瘤核组织(实验组)及其周围的正常肌组织(对照组)进行DNA片段分析;采用碘化丙啶(PI)对DNA染色,流式细胞仪检测两组细胞的凋亡率,进行细胞凋亡的定量分析.结果:实验组有9例出现DNA断裂的梯形条带(9/30,30.0%),对照组有20例出现DNA梯形条带(20/30,66.7%),二者比较有明显差异;实验组凋亡率明显低于对照组.结论:子宫肌瘤的发生与细胞凋亡异常有关.     

A case of multiple leiomyomatous lesions of the lung: An analysis of flowcytometry and hormone receptors

A 36 year old woman was admitted to our department because of a chest X-ray which showed multiple developing shadows. She underwent bilateral exploratory thoracotomies and a total 5 tumors were resected and pathologically diagnosed as benign metastasizing leiomyoma, the largest of which was positive for the progesterone receptor and negative for the estrogen receptor. A histogram of this tumor using a flow cytometer showed a diploid pattern and 4.6 percent of the S phase which was not more than that of a leiomyoma of the uterus from another patient. Two months later, she underwent a hysterectomy and bilateral salpingo-oophorectomy for treatment of the positive progesterone receptor in the pulmonary lesions. The resected uterine myoma and normal myometrium showed positive estrogen and progesterone receptors. For the subsequent 28 months she has been free of any further symptoms. Benign metastasizing leiomyoma of the uterus is a rare disease and very interesting because of its histological benignity and hormonal dependency. However, according to the literature, it is often confused in entity due to the fact that normal lung tissue also possesses hormone receptors. Considering our data on hormone receptors, it is rational to think that multiple leiomyomatous lesions in the lung should only be diagnosed as benign metastasizing leiomyomas when they possess positive estrogen and progesterone receptors.     

细小病毒H－1对正常仓鼠肝细胞DNA周期动力学的影响

健康成年金黄仓鼠腹腔内接种ＰＶＨ－１（５×１０６ｐｆｕ），ＦＣＭ动态检测肝细胞ＤＮＡ周期动力学变化，结果显示：ＰＶＨ－１接种后处于Ｇ０＋Ｇ１期的肝细胞与对照组动物相比无显著差异。Ｓ期肝细胞在第３天显著高于对照组动物（Ｐ＜０．０５），且在第７天时仍呈上升趋势，第１４天到３０天趋于正常水平。Ｇ２＋Ｍ期肝细胞在ＰＶＨ－１接种后第３０天升高（Ｐ＜０．０５）。结果提示，动物在ＰＶＨ－１接种后第３到第７天是病毒在肝细胞中的复制高峰，并阻滞仓鼠处于Ｓ期肝细胞分裂进入Ｇ２＋Ｍ期，但是这种阻滞作用较短暂、并可逆，在ＰＶＨ－１接种后１４～３０天自然消失。     

DNA flow-cytometric analysis in colorectal cancer: A comparison of metastasizing and non-metastasizing tumours

The most common cause of death in patients with colorectal cancer is metastatic liver disease. In order to identify patients at a high risk of developing hepatic secondaries from colorectal cancers, DNA content was measured in metastasizing colorectal primaries (Group I, n= 32) as well as in their subsequently resected liver secondaries and in sections of non-metastasizing colorectal cancers (Group II, n= 25). A modified interpretation system involving both a DNA index and percentage of cycling cells (those in S and G2 + M phases) was developed. DNA content was measured in paraffin-embedded sections by flow cytometry using internal controls (human peripheral blood mononuclear cells) and non-malignant tissue controls (19 patients with diverticular disease). In Group I there were significantly more tumours with both abnormal ploidy (aneuploid or abnormal tetraploid peak) and > 15% cycling cells compared with Group II (Chi-squared; P= 0.034). The combination of abnormal ploidy and > 15% cycling cells was superior to Dukes’ classification for identifying metastasizing tumours (Logistic Regression; P= 0.047). However, it was not possible to discriminate between the two groups using either DNA ploidy or the percentage of cycling cells alone. The metastasizing colorectal cancers exhibited similar DNA ploidy characteristics and had a similar percentage of cycling cells compared with their liver metastases. These results suggest that tumour DNA ploidy plus the percentage of cycling cells may predict the development of liver metastases and thus survival in patients with colorectal cancer.     

瘤体内DNA倍体异质性与食管鳞状细胞癌病理特性的关系

目的探讨食管鳞状细胞癌瘤体内ＤＮＡ倍体异质性及其与肿瘤病理学特性的关系。方法采用多点取材方法，应用流式细胞术对８０例中晚期食管鳞癌标本的ＤＮＡ倍体类型进行分析。如果一个标本中的３个样品中的倍体类型不一致，即判定为倍体异质性肿瘤（ＰＨＴ），其余为非倍体异质性肿瘤（Ｎ－ＰＨＴ）。然后用统计学方法分析ＤＮＡ倍体异质性与肿瘤病理学特性的关系。结果８０例中，共检出异倍体肿瘤７１例（８８．８％），ＤＮＡ指数范围为０．７７～１．７４。倍体异质性肿瘤的检出率为４７．５％（３８／８０）。倍体异质性与肿瘤的侵润程度，淋巴结转移及患者预后有关（Ｐ＜０．０５～０．０２５），而与组织分级和瘤体大小无关（Ｐ＞０．０５）。结论ＤＮＡ倍体异质性是一种重要的肿瘤生物学现象。深入研究它将对判断肿瘤的病理特性、患者预后及协助组织学分级具有重要的价值。     

单侧睾丸扭转引发对侧睾丸生精损伤免疫机制的实验研究

目的探讨单侧睾丸扭转对侧睾丸生精功能损伤的机理。方法雄性SD大鼠24只,随机分为3组,建立睾丸扭转动物模型。第1组为假手术组,第2、3组扭转左侧睾丸720°并固定维持,分别于12 h和24 h后复位固定。术后1个月留取对侧睾丸称重。应用流式细胞术(FCM)检测各组生精细胞凋亡和各级生精细胞计数,免疫组化SABC法检测抗精子抗体免疫复合物(IgG)在睾丸组织中的沉积。结果第1、2、3组对侧睾丸重量分别为(1555.73±72.34)、(1184.20±101.02)、(783.60±117.93)mg,凋亡细胞数依次为53.25±8.61、1622.00±129.31、3401.25±179.75,抗精子抗体阳性染色率为0、0.55±0.02、0.69±0.03,组间两两比较差异均有统计学意义(P<0.01)。与第1组相比,第2、3组对侧睾丸单倍体、二倍体和四倍体细胞群计数明显减少,同时可见抗精子抗体免疫复合物在睾丸组织中明显沉积,其差异均有统计学意义(P<0.01)。第2、3组间(除四倍体细胞群计数外)差异亦有统计学意义(P<0.01)。结论单侧睾丸扭转导致对侧睾丸生精细胞广泛凋亡,自体免疫反应可能在对侧睾丸生精功能损伤中起重要作用。     

高糖等因素对系膜细胞细胞周期调节负控蛋白P27的影响

目的探讨高糖不同浓度刺激或抑制剂对肾小球系膜细胞表达细胞周期负控蛋白P27的影响.方法不同浓度高糖(5.5 mmol/L和25mmol/L、内皮素-1(10-7mol/L)、白介素-13(10 ng/ml和100 ng/ml)作用于培养的系膜细胞.流式细胞仪检测系膜细胞P27表达水平.结果5.5mol/L浓度组在不同时间P27表达为5.10±0.94和3.84±0.81,较对照级明显降低(P＜0.001),而25 mmol/L浓度组在不同时间P27表达为26.82±3.15和30.88±3.68,较对照组明显升高.内皮素-1刺激14 h和18 h后P27的表达分别为14.76±1.49和12.18±1.30,与对照组比较有明显差异(P＜0.05,P＜0.05).IL-13 10 ng/ml浓度组和100 ng/ml浓度组P27表达为46.74±3.52和23.8±2.56,对对照组比较有明显差异(P＜0.001,P＜0.05),不同浓度组之间有显著差异(P＜0.01).结论细胞周期调节负控蛋白P27在调节系膜细胞增殖过程中起重要作用.     

彩色多普勒超声对肝癌肝动脉化疗栓塞的疗效评价

目的 :探讨彩色多普勒超声 (CDFI)在评价肝癌肝动脉化疗栓塞 (TAE)疗效中的应用价值。方法 :对 4 8例原发性肝癌TAE术前、后行CDFI检查 ,观察肿瘤大小、血供及肝动脉、门静脉血流动力学改变。结果 :TAE术后大多数肿瘤均有不同程度缩小 ;肿瘤内部及周边血供明显减少 (P <0 0 5 ) ,以肿瘤内血流减少更明显 ,其血供减少程度与肿瘤缩小程度呈正相关 (r =0 75 ,P <0 0 0 1) ,肝动脉峰值流速明显下降 ,门静脉血流速度增快 (P <0 0 0 1)。结论 :CDFI能客观判断肿瘤大小及血流灌注状态 ,是目前检验肝癌TAE效果较理想的影像学检查方法