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1.
从鼠的肝脏组织中提取总 RNA,采用 RT- PCR获得了鼠层粘蛋白 ( laminin)α5链的 E3 ( L G4 - 5)、L G4 和 L G5等 3个基因片段 ,与 NCBI数据库参考序列相比 ,同源性在 99%以上。将 E3 ( L G4 ,5)、L G4 和 L G5定向克隆到原核高效表达载体 Pet- 2 8a中 ,构建 Pet- 2 8a- E3 、Pet- 2 8a- L G4 、Pet- 2 8a- L G5等 3个原核表达质粒。将表达质粒转化表达受体菌 BL2 1中 ,IPTG诱导表达。收集菌液进行 SDS- PAGE电泳、Western- blotting分析 ,结果显示 ,这 3段基因在原核细胞中成功表达 ,薄层扫描显示表达蛋白占细胞总蛋白的 2 0 %以上。大量提取包涵体 ,纯化后免疫家兔 ,8周后得多克隆抗体 ,间接 EL ISA在抗体稀释到 1∶ 1 2 80 0时仍为阳性 相似文献
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ABSTRACT: This study incorporated the 43 kDa Zn-binding membrane protein isolated from common carp into liposome. The specificity and strength of the binding of 65 Zn to the 43 kDa protein-liposomes, and the binding of the 65 Zn-labeled 43 kDa protein-liposomes to laminin were studied. It was found that 65 Zn was bound to the external side of the 43 kDa protein-liposomes. Specific binding of 65 Zn to the protein-liposomes was detected. The binding parameter of Zn to the protein was found to be: maximum binding site (Nmax ), 76.7 pmole/µg protein (approx. 3 mole of Zn2+ /mole); and equilibrium dissociation constant (Kd), 0.19 µM. Of the cations introduced (Ca2+ , Cd2+ ,Co2+ , Cr2+ , Cu2+ , Fe2+ , Hg2+ , Mg2+ , Mn2+ , Ni2+ , Pb2+ ), only Co2+ competed significantly with Zn. The protein-liposomes were also found to bind specifically to laminin with a Nmax of 1.1 pmole/µg laminin, and Kd of 4.79 µM. No significant protein-liposome binding occurred to other extracellular matrix proteins (fibronectin, fibrinogen or vitronectin). Furthermore, the binding was specifically inhibited by the Arg-Gly-Asp (RGD) peptide or GRGDSPG, while two other analogs (GRGESPG and GRADSPG) were without effect. 相似文献
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Objective:To observe the effect of Busui Shengxue Granule (补髓生血颗粒Herbal granule for replenishing marrow to produce blood) on chronic aplastic anemia (CAA) patients' integrin a 6 (VLA-6/CD49f) and... 相似文献
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目的 :检测肺癌患者血浆 P-选择素 (P- selectin,P- sel)和层粘连蛋白 (laminin,L N)的水平并探讨其临床意义。方法 :用 EL ISA法检测了 85例不同期肺癌患者血浆中 P- sel和 L N的含量。结果 :各期肺癌患者血浆中 P- sel和 L N与正常对照组比较差异有显著性 (P<0 .0 1) ;恶性程度较高的小细胞性肺癌患者血浆 L N与非小细胞性肺癌比较差异有显著性 ,但两者 P- sel差异无显著性 ;小细胞性肺癌中广泛型患者血浆 P- sel和 L N与局限型比较差异有显著性 (P<0 .0 1) ;非小细胞性肺癌患者中 期与 期、 期比较 ,血浆 P- sel和 L N的含量的差异均有显著性 (P<0 .0 1) ;18例小细胞性肺癌患者经化疗后随肿瘤体积的缩小和临床症状的改善 ,P- sel和 L N水平较化疗前显著降低。结论 :肺癌患者血浆 P- sel和L N的含量随病情的恶化而显著增高。检测血浆 P- sel和 L N对肺癌患者的病情判断及疗效评估有一定的参考价值。 相似文献
5.
The hepatoprotective effects of polyprenols from Ginkgo biloba L. leaves were evaluated against carbon tetrachloride induced hepatic damage in Sprague-Dawley rats. The elevated levels of serum ALT, AST, ALP, ALB, TP, HA, LN, TG, and CHO were restored towards normalization significantly by GBP in a dose dependent manner. The biochemical observations were supplemented with histopathological examination of rat liver sections. Meanwhile, GBP also produced a significant and dose-dependent reversal of CCl4-diminished activity of the antioxidant enzymes and reduced CCl4-elevated level of MDA. In general, the effects of GBP were not significantly different from those of the standard drug Essentiale. 相似文献
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目的：探讨皮肤角质形成细胞肿瘤中Ⅳ型胶原和层粘蛋白的表达及其与肿瘤转移的关系。方法：用ABC免疫组化技术观察48例皮肤鳞状细胞癌、40例基底细胞癌及l5例角化棘皮瘤手术切除标本。结果：Ⅳ型胶原和层粘蛋白在高分化或无淋巴结转移的鳞状细胞癌中的表达明显高于低分化或有淋巴结转移者(P＜0．01)。基底细胞癌中Ⅳ型胶原和层粘蛋白的表达类似于鳞状细胞癌(P＞0．05)，但其表达明显高于有转移的鳞状细胞癌(P＜0．01）。角化棘皮瘤中Ⅳ型胶原和层粘蛋白的表达一般呈连续性，未见明显缺乏。结论：皮肤癌的癌分化、淋巴结转移与Ⅳ型胶原和层粘蛋白的表达有关。 相似文献
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旨在研究Ghrelin对产蛋鸡等级前卵泡发育的调节作用.通过RT-PCR方法检测卵泡Ghrelin受体(GH-SR)的表达情况,采用组织学和免疫组化方法探讨Ghrelin和促卵泡素(FSH)单独或联合处理对鸡等级前卵泡形态学的变化以及层粘连蛋白(LN)和缝隙连接蛋白43(Cx43)的标记率.结果表明,GHSR在颗粒层和膜层上均有表达,其表达量随着卵泡的发育分别至大白卵泡(LWF)和小黄卵泡(SYF)达到最高,随后逐渐降低.悬浮培养的卵泡经Ghrelin和FSH处理24 h后,LWF和SYF的颗粒层和膜层厚度及颗粒细胞密度均显著增加(P＜0.05).同时,颗粒细胞中LN和Cx43的标记率显著提高(P＜0.05),且以Ghrelin与FSH联合处理组最为显著.结果提示,Ghrelin可通过提高LN和Cx43的表达以增强颗粒细胞的连接功能,促进鸡等级前卵泡颗粒细胞的增殖,并可协同FSH调节卵泡的发育. 相似文献
8.
Cutaneous myxidiosis in European eel,Anguilla anguilla (Linnaeus, 1758): histopathology,histochemistry and laminin immunohistochemistry 
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下载免费PDF全文 Histopathology, histochemistry and immunohistochemistry of the integument of European eel, Anguilla anguilla (Linnaeus, 1758), infected by Myxidium sp. are reported. Skin samples from affected and unaffected eels were dissected, formalin fixed, paraffin embedded, sectioned and stained with H&E, Periodic acid–Schiff's staining method, Alcian Blue 8 GX pH 2.5/Periodic acid–Schiff's and McCallum‐Goodpasture's Gram stain. Moreover, immunohistochemistry was performed using a primary polyclonal laminin antibody. Histologically, cysts (diameter 2–3 mm) were observed mainly under the scale pockets, encircled by a thin collagen layer, lined by elongated, flattened fibroblasts and containing bipolar, PAS‐ and Gram‐positive spores with opposite polar capsules. The epidermis stretched by the underlying cyst appeared dysplastic, thinned with a significant reduction in mucous cells number. Only inconsistent and aspecific inflammatory reaction was noted around the cysts at the dermis/epidermis interface. Intense laminin‐like protein immunolabel was documented in the plasmodial ectoplasm and related to host anergia. This was the first report of laminin immunolabel in a member of the Myxozoa. Epidermal dysplasia represents likely an aspecific response against the underlying tensile force exerted by the developing parasite cyst, while fibroblast and collagen encapsulation denote a parasite‐driven host response protecting, rather than harming, the encircled parasite. 相似文献
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Primary cell culture and morphological characterization of canine dermal papilla cells and dermal fibroblasts 总被引：12，自引：0，他引：12
Bratka-Robia CB Mitteregger G Aichinger A Egerbacher M Helmreich M Bamberg E 《Veterinary dermatology》2002,13(1):1-6
Skin biopsies were taken from female dogs, the primary hair follicles isolated and the dermal papilla dissected. After incubation in supplemented Amniomax complete C100 medium in 24-well culture plates, the dermal papilla cells (DPC) grew to confluence within 3 weeks. Thereafter, they were subcultivated every 7 days. Dermal fibroblast (DFB) cultures were established by explant culture of interfollicular dermis in serum-free medium, where they reached confluence in 10 days. They were subcultivated every 5 days. For immunohistochemistry, cells were grown on cover slips for 24 h, fixed and stained with antibodies against collagen IV and laminin. DPC showed an aggregative growth pattern and formation of pseudopapillae. Intensive staining for collagen IV and laminin could be observed until the sixth passage. DFB grew as branching, parallel lines and showed only weak staining for collagen IV and laminin. 相似文献
10.
Transmissible spongiform encephalopathies known as prion diseases are a group of fatal neurodegenerative disorders that affect both humans and animals. The generally accepted principle of the disease is that the conversion of the cellular prion protein (PrP(c)) into the disease associated isoform PrP(Sc) leads to spongiform degeneration of the brain and amyloid plaque formation. Until now no therapy leading to potential alleviation or even cure of the disease exists. It is therefore important to develop therapeutic approaches for the treatment of TSEs since these infections are inevitably fatal and, especially in the case of vCJD, they affect youngsters. Besides current conventional therapeutic strategies, this review summarizes new therapeutic tools targeting the prion receptor LRP/LR. 相似文献