人大肠癌中Ⅳ型胶原酶与基底膜免疫组织化学双重染色研究

目的观察Ⅳ型胶原酶和基底膜在大肠癌生长中的相互关系及Ⅳ型胶原酶的表达与ｐ２１的表达之间的关系。方法应用免疫组化双标记方法，对８６例人大肠癌组织的Ⅳ型胶原酶表达及基底膜的改变作对照研究。结果大肠癌组织中Ⅳ型胶原酶的阳性率为８３．７２％（７３／８６），与对照组癌周正常肠粘膜的阳性率１０％差异有非常显著意义（Ｐ＜０．０１）。同时，双染色显示Ⅳ型胶原酶阳性的癌组织基底膜表现为不连续，出现缺口、片段缺失或完全消失等形态。此外，肠癌中Ⅳ型胶原酶的分布与ｐ２１的表达呈正相关（ｒ＝０．９７４，ｐ＜０．０１）。结论Ⅳ型胶原酶的分泌对肠癌基底膜破坏起重要作用，可以作为判断大肠癌生长特性的辅助指标。     

单侧外周伤害性刺激诱发大鼠脊髓中转录因子CREB的双侧磷酸化(英文)

c AMP反应成分结合蛋白 (c AMP response elem ent-binding protein,CREB)是一种转录因子 ,它在磷酸化之后可调节靶基因的转录。 CREB在 13 3位置的丝氨酸的磷酸化 ,与脊髓中伤害性传入的处理有关 ,本文作者等用特殊抗体对此进行了免疫细胞化学研究。在正常大鼠 ,虽然几乎所有脊髓神经元的核中都可见 CREB的轻度着色 ,但磷酸化的 CREB仅见于双侧腰段脊髓的 ～ 层 (75± 15 vs60± 18)和 层 (9± 3 )。用福尔马林注射引起一侧后脚掌出现炎症时 ,可在双侧腰段脊髓看到磷酸化CREB细胞核的快速 (小于 5 min)和节段性的显著增多 ;它们主要分布在双侧背角表层 ～ 层 (2 5 4± 2 0 vs 2 62± 2 3 )、 层(115± 13 )和双侧 ～ 层 (3 46± 2 0 vs3 2 8± 2 6) ;而在对照和炎症组大鼠的胸段脊髓中均未见磷酸化 CREB的增加。在注射CFA诱发一侧炎症或切断一侧坐骨神经的实验组大鼠 ,也可看到至少延续到第三天的强而双侧性的 CREB的磷酸化。这种由一侧后肢伤害性传入引致腰段脊髓中镜像式双侧 CREB磷酸化的出现 ,与一般看到的损伤传入只在同侧脊髓背角引起某些神经化学改变的结果不同 ,可能是人神经损伤后或在实验动物中出现对侧镜像式疼痛过敏现象的基础     

An IFN-beta-albumin fusion protein that displays improved pharmacokinetic and pharmacodynamic properties in nonhuman primates.

The long half-life and stability of human serum albumin (HSA) make it an attractive candidate for fusion to short-lived therapeutic proteins. Albuferon (Human Genome Sciences [HGS], Inc., Rockville, MD) beta is a novel recombinant protein derived from a gene fusion of interferon-beta (IFN-beta ) and HSA. In vitro, Albuferon beta displays antiviral and antiproliferative activities and triggers the IFN-stimulated response element (ISRE) signal transduction pathway. Array analysis of 5694 independent genes in Daudi-treated cells revealed that Albuferon beta and IFN-beta induce the expression of an identical set of 30 genes, including 9 previously not identified. In rhesus monkeys administered a dose of 50 microg/kg intravenously (i.v.) or subcutaneously (s.c.) or 300 microg/kg s.c., Albuferon beta demonstrated favorable pharmacokinetic properties. Subcutaneous bioavailability was 87%, plasma clearance at 4.7-5.7 ml/h/kg was approximately 140-fold lower than that of IFN-beta, and the terminal half-life was 36-40 h compared with 8 h for IFN-beta. Importantly, Albuferon beta induced sustained increases in serum neopterin levels and 2',5' mRNA expression. At a molar dose equivalent to one-half the dose of IFN-beta, Albuferon beta elicited comparable neopterin responses and significantly higher 2',5'-OAS mRNA levels in rhesus monkeys. The enhanced in vivo pharmacologic properties of IFN-beta when fused to serum albumin suggest a clinical opportunity for improved IFN-beta therapy.     

21－羟化酶CYP21B基因Ile~（172）→Asn错义突变

为了解先天性肾上腺皮质增生症患者的２１－羟化酶ＣＹＰ２１Ｂ基因中Ｉｌｅ~（１７２）→Ａｓｎ错义突变的发生率，根据放大受阻突变体系（Ａｍｐｌｉｆｉｃａｔｉｏｎｒｅｆｒａｃｔｏｒｙｍｕｔａｔｉｏｎｓｙｓｔｅｍ，ＡＲＭＳ）的要求，设计了３种引物：５＇ｄ（ＴＴＧＧＧＡＧＡＣＴＡＣＴＣＣＣＴＧＣＴＣＴ）３＇（共同引物）、５＇ｄ（ＡＧＧＴＧＡＧＧＴＡＡＣＡＧＡ）３＇（正常引物）、５＇ｄ（ＡＧＧＴＧＡＧＧＴＡＡＣＡＧＴ）３＇（突变引物），在７例患儿中进行了检测，发现具有本突变者３例。对其中一例进行的家系分析，结果提示：这组引物有快速、简便的优点，不需使用同位素就能对具有Ｉｌｅ~（１７２）→Ａｓｎ变异的高危家庭成员作产前诊断。     

Functional CD4+T cell subsets defined by expression of CD45RC and NTA260 antigens and age-associated polarization in murine lupus

Using two mAb, one specific to the alternative exon 6-dependentepitope of CD45 molecules(JH6.2) and one a natural thymocytotoxicautoantibody (NTA) with an unknown reactive epitope (NTA260),we subdivided splenic CD4⁺ T cells from 2-month-old BALB/c miceinto five phenotypically distinct subsets. CD45RC⁺NTA260^–(SI) cells were phenotypically analogous to CD4⁺ T cells predominatingin newborn mice and produced a significant amount of IL-2, butnot so IL-4, IL-10 or IFN- when stimulated with immobilizedanti-CD3 mAb in vitro. They appeared to consist mainly of naiveT_hP cells. The CD45RC⁺;NTA260⁺ (S II) subset also produced IL-2,but not other cytokines; however, the IL-2 levels produced weremuch higher than seen with the S I subset, thereby suggestingthe predominance of further maturated T_hP cells. The D45RC^–NTA260⁺(S III) subset mainly produced IL-4, IL-10, IFN- and less IL-2,and contained memory cells that helped the secondary antibodyresponse to a recall antigen, and hence contained T_h2 and probablya mixture of T_h0 and T_h1 cells. The CD45RC^–NTA260^–(S IV) subset was a poor responder to the immobilized anti-CD3mAb. The CD45RC^brightNTA260^dull(S V) subset consisted of a smallnumber of cells that were phenotypically analogous to activatedCD4⁺ T cells. While an age-associated decrease in the proportionof S I and less markedly in S II and in turn increase in S IIIsubsets of CD4⁺ T cells occurred in normal BALB/c mice, autoimmunedisease-prone (NZBxNZW)F₁ mice showed a marked age-associateddecrease in the proportion of not only S I, II but also IIIsubsets. As aged (NZBxNZW)F₁ mice carry CD4⁺ T helper cellsfor IgG anti-DNA antibody production, such age-associated polarizationto the S IV subset appears to be critical in the pathogeneslsof autoimmune disease in these mice.     

Use of radionuclide scanning to estimate size of spleen in vivo.

Splenic ultrasound was performed and spleen scans obtained by a rectilinear scanner after injection of In-113m-labelled heat damaged red cells on 14 patients with various haematological disorders. Correlation was assessed between the measured volume of the operatively removed spleens, their volumes as calculated from the ultrasound data, and their preoperative dimensions of length, circumference, and area on the scans. The relation of scan to volume was best defined by an equation V = 9.88A - 534, where V = volume of spleen (cm3) and A = area (cm2) on posterior scan. The mean percentage difference between calculated and actual spleen volume was 0.2 (SD 6.7)%. Average spleen density was established as 1.04 g/cm3. The formula was shown to be reliable in the range of splenic area c 73-450 cm2--that is, volume c 185-4000 cm3. This range includes the average normal sized spleens and the splenomegalies that are most often found in clinical practice. The formula is not valid when the spleen is smaller than normal. As the method depends on measurement of the spleen image shown by scintigraphs, it cannot be used when there is functional asplenia.     

小白鼠腘淋巴结淋巴窦的超微结构

我们观察了幼年小白鼠腘淋巴结淋巴窦的超微结构,窦壁由腔面向外一般出3层构成:(1)一层连续的内皮;(2)由一薄层细胞间质构成的基膜;(3)一层外膜网状细胞及其扁平的突起。内皮细胞核扁圆,异染色质细小,核仁不明显,胞质极薄,含粗面内质网极少,而有大量的吞饮小泡。细胞邻接处相互重叠或嵌合,有20nm宽的间隙相隔。有的地方可见不发达的细胞连接。当巨噬细胞或淋巴细胞穿越内皮时,可出现临时性间隙,内皮边缘与穿越细胞相贴。基膜由电子透明的无定形基质及细的胶原原纤维构成。外膜网状细胞的核异染色质较多,核仁明显,胞质丰富,含许多粗面内质网而很少吞饮小泡。外膜网状细胞之间常见0.5μm宽的间隙。在窦腔内有巨噬细胞、淋巴细胞及星状的网状细胞等。由于小白鼠腘淋巴结的淋巴窦小,故窦内星形的网状细胞很少,其超微结构特点,与外膜网状细胞及淋巴组织内的网状细胞相似,在突起的切面上,常见包有小束的胶原原纤维。巨噬细胞较多,形态不规则,常贴附于内皮细胞表面,核椭圆,常有凹陷,胞质丰富,含许多溶酶体。在取材前1h,于足垫注射中国墨汁的小鼠,巨噬细胞吞噬了大量粗的墨汁颗粒。本文认为,淋巴窦壁可能具有一定的屏障作用。     

Immunological heterogeneity of autoreactive T lymphocytes against the nicotinic acetylcholine receptor in myasthenic patients

The response of human T lymphocytes against the nicotinic acetylcholine receptor (AChR) was studied in five patients with myasthenia gravis (MG) and in six healthy donors using either native Torpedo AChR or recombinant protein derived from the mammalian AChR α subunit (X4, residues 6–216 of mouse AChR α subunit). The present study demonstrates that (a) AChR-specific T helper cell lines can be generated from MG patients [either from peripheral blood lymphocytes (PBL) or from thymocytes] as well as from PBL of normal controls, (b) lymphocytes from MG patients, but not from controls, recognize the mammalian AChR but not the Torpedo receptor, (c) in humans, the HLA-DR2-associated T cell epitope is probably located in the region of residues 162–216 of the AChR α subunit and (d) there is a considerable heterogeneity of autoreactive T cell responses: (i) Tcell lines from different HLA-type donors have distinct epitope profiles; (ii) the epitope specificity of the PBL-derived T cell line is different from that of the thymocyte-derived line; (iii) the epitope specificities of patient-derived T cell lines are different from those generated from normal controls who share the same HLA phenotype.     

Study on structure-property relationship of polyimide blends, 2. Structure and miscibility of polyimide PBPI-E/PTI-E blends

The structure and miscibility of polyimide PBPI-E/PTI-E blends were studied by wide- and small-angle X-ray scattering and dynamic mechanical analysis, where PBPI-E is a biphenyldianhydride-based polyimide, and PTI-E is a polyimide from 4,4′-thiodiphthalic anhydride and 4,4′-oxydianiline. The results obtained show that there exists a paracrystalline structure in the blends with high content of PBPI-E, but this does not affect the miscibility of the blends. The blends are miscible over the entire composition range, since only one T_g was observed for each blend. Meanwhile, the segregation of PTI-E during crystallization of PBPI-E in the blends is interlamellar.