大鼠高氧暴露导致的肺泡内皮细胞结构和功能的变化(论著)

目的：进一步探索肺氧中毒的机理。方法：在大鼠常压高浓度氧（97％±2％）暴露的早期，观察肺血管紧张素转化酶（ACE）活性、脂质过氧化物（LPO）水平及肺泡毛细血管超微结构的变化。结果：随着肺ACE活性的下降和LPO水平的升高，部分肺泡内皮细胞（EC）出现胞浆水肿，同时肺泡毛细血管内白细胞、血小板明显增多。结论：肺泡EC胞膜功能的受损，尤其是胞膜结合的ACE活性下降所致对炎性介质清除的减少，可能起动了以肺泡EC胞膜通透性增高和炎细胞粘附为特征的早期肺氧中毒。     

Analysis of T Cell Receptor Variability in Fresh Tumor-infiltrating Lymphocytes from Human Head and Neck Cancer

In this study, we analyzed T cell receptor (TCR) gene rearrangements in tumor-infiltrating lymphocytes (TIL) freshly obtained from 15 patients with head and neck cancer using the reversely transcribed polymerase chain reaction (RT-PCR) method. These TILs showed preferential expression of Vα10, Vα8 and Vα1, detected in 13 (87%), 11 (73%), and 9 cases (60%), respectively. The TCRVβ gene revealed diversity without preferential usage. The head and neck region is exposed to bacteria and viruses, so it is possible that the tumor site can become infected and accumulate T cells involved in infection and inflammation. Therefore, we also investigated TCR gene usage in T cells infiltrating in chronic sinusitis mucosa to address the question of whether the Vα1, Vα8, and Vα10 subfamilies are characteristic in TIL from squamous cell carcinoma of head and neck. TCR Vα10 gene usage was also the most common in Vα segment in T cells infiltrating the sinus mucosa, but Vα and Vα8 were not detected in the T cells in sinusitis. These results indicate that the Vα10 subfamily, the preferred T cell population in both TIL and T cells in inflammatory disease, might he involved mainly in inflammation or infection. On the other hand, Vα1 and Vα8 appear to be relatively specific populations for antitumor immunity in head and neck cancer.     

Vein of Galen and sinus thrombosis with bilateral thalamic infarcts in sickle cell anaemia: CT follow-up and angiographic demonstration

A 2-year-old boy with known sickle cell disease presented in acute coma. CT revealed bilateral thalamic infarcts and incomplete sinus thrombosis. Angiography confirmed thrombosis of the straight sinus and vein of Galen.     

T cell responses to orbital antigens in thyroid-associated ophthalmopathy.

Thyroid-associated ophthalmopathy (TAO) is most likely to be a T cell-mediated disease, in which cytokines released in the extraocular muscles activate fibroblasts, increasing glycosaminoglycan production. The nature of the orbital antigen recognized by the infiltrating T cells is unclear, although it is possible that there is cross-reactivity between this and a thyroid autoantigen to explain the close association with thyroid autoimmunity. We have tested the ability of human and porcine eye muscle antigen preparations to stimulate proliferation of circulating T cells from healthy subjects and patients with TAO or Graves' disease without clinical TAO. Occasional responses were seen, particularly after depletion of CD8+ T cells, and two out of 10 TAO patients responded to eye muscle proteins of 25-50 kD after fractionation of antigens on gels and subsequent elution. There was no disease-specific response of T cells to R1, R14, D1 and 1D3, recombinant proteins identified from screening an eye muscle cDNA library with sera from patients with autoimmune thyroid disease. We have also found that interferon-gamma (IFN-gamma) production by T cells from TAO patients was not stimulated by eye muscle membrane antigens or by 1D3. These results suggest that the frequency of circulating T cells responding to eye muscle antigens in TAO is low, and that several candidate orbital antigens, including the 64-kD protein 1D3, are unlikely to be important T cell autoantigens in this condition.     

应用因特网对SARS病毒M蛋白B细胞抗原表位的预测

目的：预测SARS冠状病毒M蛋白的B细胞表位。方法：采用EMBOSS软件结合Hopp＆woods亲水性参数、Janin可及性参数、极性参数、柔韧性参数及二级结构方案对SARS冠状病毒M蛋白的B细胞表位进行了预测，并用吴玉章等已建立的B细胞表位预测方法进行评述。结果：SARS冠状病毒M蛋白B细胞识别的表位可能位于第3～13和153～166位氮基酸等区域内或附近，这2个被预测的表位均含有β—转角和无规卷曲结构。结论：该研究对应用合成肽抗原进行SARS早期诊断研究及相关抗体的制备具有重要指导意义。     

缬沙坦与苯那普利拉对SHR肾小球系膜细胞的保护作用

目的：观察ARB－valsartan及ACEI－benazeprilat能否抑制由AngⅡ、PDGF引起的SHR肾小球系膜细胞的增殖与肥大。且比较单用时作用孰强孰弱，联合应用后有无协同效应。方法；采用经典SHR系膜细胞培养技术，细胞中加入各种因子和（或）药物，以^3H-leucine或^3H-thymidine掺入后的cpm值反映蛋白或DNA合成。采用t test检验差异的显著性。结果：1、系膜细胞在AngⅡ、PDGF刺激后引起的增殖与肥大，均可被valsartan及benazeprilat抑制。2、比较发现单一用药在同一浓度下，valsartan较benazeprilat能更有效地抑制细胞的增殖与肥大。3、联合用药在同一浓度valsartan benazeprilat均比单用valsartan或benazeprilat效果明显。结论：1、valsartan与benazeprilat均能抑制SHR系膜细胞的增殖与肥大。2、valsartan在抗系膜细胞增殖与肥大作用稍强于benazeprilat。3、在一定浓度条件下联合用药对抗系膜细胞的增殖与肥大作用，较单一用药效果明显。     

骨髓间充质干细胞-支架复合体向软骨诱导分化的实验研究

目的:以骨髓间充质干细胞作为种子细胞,探索组织工程化软骨的构建.方法:分离、获取、扩增骨髓间充质干细胞,通过将扩增的骨髓间充质干细胞接种于聚羟基乙酸(PGA)上,在成软骨诱导因子作用下在体外诱导培养3周后,将实验组和对照组的标本转植入自体兔腹腔内,进行体内培养6～12周后,再进行检测,评估工程化软骨的形成.结果:骨髓间充质干细胞接种于PGA支架上后,经过体外诱导培养和体内培养后,获得标本(7/10)出现软骨组织外观,进行组织学切片可见软骨陷窝,进行Ⅱ型胶原蛋白的免疫组化、Ⅱ型胶原mRNA原位杂交均为阳性.结论:兔骨髓间充质干细胞在成软骨诱导剂作用下,经体外和体内培养后,可生成组织工程化类软骨.     

纤维蛋白胶体外重建兔角膜上皮的实验研究

目的利用组织工程技术体外重建角膜上皮组织,为重建角膜表面提供良好的移植材料和方法。方法以等量凝血酶和纤维蛋白原作用生成的纤维蛋白胶为载体(实验组),种植兔角膜缘组织块,体外培养重建角膜上皮层,每天记录细胞生长覆盖面积,从生长特性、光镜特征、超微结构特点以及免疫细胞化学方面进行观察检测。结果角膜缘上皮细胞在纤维蛋白胶上黏附生长增殖,体外培养7d左右即形成单层上皮细胞,2周左右形成的复层上皮细胞与纤维蛋白胶构成角膜上皮组织经检测与生理状态下的角膜上皮组织相近。实验组与对照组(置盖玻片为载体)比较细胞生长覆盖面积第4天至第7天差异有显著性(P<0.05)。结论以纤维蛋白胶为载体,体外培养的角膜上皮组织片可作为眼表重建角膜上皮移植良好的来源。     

HIGH-LEVEL EXPRESSION OF RATD1A DOPAMINE RECEPTOR cDNA IN MOUSE FIBROBLASTLTK- CELLS BY n-BUTYRATE

1. In orefer to develop a simple, efficient system for the highlvel expression of dopamine recetors in eukaryotic cells, we have studied the effects of n-butyrate on the expression of rat D_1A dopamine receptor cDNA in mouse fibroblast LTK- cells as compared with those of n-butyrate on endogenous D₁ receptor levles in opossum kidney cells.
2. In the transfected LTK- cell mebranes with pRc/CMVD_1A receptor cDNA, a selective D₁ dopamine antagonist, [³H]-SCH 23390, exhibited a K_d of 0.9 ± 0.1 nmol/L and a B_max of 0.35 ± 0.05 pmol/mg protien (n = 5).
3. Addtion of n-butrate (2–10 mmol/L) to the culture medium for 48h dose-dependently increased the D_1A receptor level up to 1.5 ± 0.3 pmol/mg protien (n = 7), although the K_d values were not affected. The increase in receptor level was accompanined by an elevation of selective D₁ agoinist-induced adenyly cyclase activity.
4. In contrast, n-butyrate treatment (2–10 mmol/L) did not affect either endogenous D₁ receptor levels or fendoldopmainduced adenylyl cyclase activity in possum kidney cells.
5. These results sugges n-butyrate is a sueful tool for obtaining high-level expression of D_1A dopamine receptor cDNA in mouse fibroblast LTK- cells.