羊驼皮肤角蛋白基因家族的表达

为了揭示羊驼皮肤结构发生和皮肤生理的分子机制,对构建的皮肤cDNA文库进行了大规模的测序,以对羊驼皮肤的结构基因--角蛋白基因家族进行基因水平的表达分析.通过构建羊驼皮肤cDNA文库的基因表达谱,发现了羊驼皮肤结构基因角蛋白的表达具有以下特点:keratin家族有7个家庭成员表达,其中上皮角蛋白类型I keratin10表达丰度最高,而上皮角蛋白类型Ⅱ表达丰度较低;毛干特异性上皮角蛋白以类型I keratin 25低表达为主;毛发角蛋白以类型I keratin 40低表达为主.这些基因的表达对维持羊驼皮肤及其衍生物(毛发)的正常生理起着重要的作用.     

肝龙胶囊联合水飞蓟宾胶囊对肝纤维化大鼠的治疗作用

观察肝龙胶囊联合水飞蓟宾胶囊对肝纤维化大鼠的治疗效果,探讨肝龙胶囊是否对水飞蓟宾胶囊抗肝纤维化具有增效的作用。用猪血清制备大鼠肝纤维化模型,水飞蓟宾胶囊临床给药剂量单用及联合不同剂量肝龙胶囊处理肝纤维化大鼠,通过试剂盒及酶联免疫吸附试验(ELISA)检测大鼠血清和肝组织内肝纤维化标志物透明质酸(hyaluronic acid,HA)、层黏素(laminin,LN)、血清Ⅲ型前胶原氨基端肽(procollagenⅢN-termi,PCⅢ)、Ⅳ型胶原(typeⅣcollagen,Ⅳ-C)及转化生长因子β1(transforming growth factor-β,TGF-β1)、Ⅰ型胶原(collagen typeⅠ,Col-Ⅰ)的含量,以及肝脏内丙二醛(malondialdehyde,MDA)、谷草转氨酶(glutamic oxaloacetic transaminase,AST)、谷丙转氨酶(Cereal third transaminase,ALT)、γ-谷氨酰转肽酶(γ-glutamyl transpeptidase,γ-GT)的含量。与模型组比较,各治疗组中大鼠血清肝纤维化标志物的含量均低于模型组(P<0.05);肝龙胶囊与水飞蓟素联合用药与单用水飞蓟宾组相比,可使大鼠血清内HA和TGF-β1的含量显著降低(P<0.05);大鼠肝脏组织中除PCⅢ外,其余血清肝纤维化标志物的含量治疗组均低于模型组。肝组织中的γ-GT和ALT高剂量联合给药组显著降低,AST无显著差异,MDA中剂量联合给药组显著升高。说明肝龙胶囊可增强水飞蓟宾胶囊对猪血清性肝纤维化大鼠的抗肝纤维化作用。     

Agouti在不同颜色被毛羊驼皮肤组织中的表达

哺乳动物毛色是一个多基因控制的性状,Agouti是研究哺乳动物毛色发生的一个主要的候选基因。为研究Agouti基因在羊驼毛色发生过程中是否表达及其作用机理如何,本研究分别采用半定量RT-PCR技术和免疫组化技术对白色和棕色被毛成年羊驼皮肤组织进行研究。半定量RT-PCR显示,Agouti基因在白毛羊驼皮肤组织中高量表达,而在棕色毛羊驼皮肤组织中表达量较低;免疫组织化学显示,棕色被毛羊驼皮肤组织毛囊外结缔组织中有少量的Agouti阳性着色,白色羊驼毛囊内根鞘有大量的Agouti阳性着色,且形成了集中的Agouti特异性阳性着色圈。研究结果说明:Agouti基因在不同被毛羊驼皮肤组织中均有表达,在白色被毛羊驼真皮毛乳头表达的Agouti基因可能与白色被毛发生相关。     

黑素皮质素受体1(MC1R)在不同毛色羊驼皮肤组织中的表达与定位研究

旨在通过研究黑素皮质素受体1(MC1R)在羊驼皮肤组织中的定位与表达,探讨其在羊驼毛色形成中的作用机制及与毛色的相关性。选用成年白色羊驼与棕色羊驼为研究对象,采用免疫组织化学方法及免疫印迹法(Western blotting),对MC1R在羊驼皮肤中的表达进行定位和定量分析。结果,(1)免疫组化结果显示,MC1R蛋白在白色和棕色羊驼皮肤表皮、毛囊周围外根鞘组织、毛乳头以及毛球周围都呈阳性着色。在棕色羊驼,阳性表达部位主要分布于毛球顶部及表皮,呈强阳性着色。在白色羊驼,阳性表达部位主要在外根鞘及表皮,毛球部表达呈弱阳性。MC1R在棕色羊驼皮肤组织中极显著表达(P<0.01)。(2)Western blotting结果显示,羊驼皮肤组织粗蛋白提取物中存在分子量约35ku与兔抗MC1R多克隆抗体发生免疫阳性反应的蛋白条带,且棕色羊驼平均蛋白表达量显著高于白色羊驼,差异极显著(P<0.01)。MC1R在羊驼皮肤组织中的定位与表达量的不同与羊驼毛色表型之间存在一定的相关性。     

3β-HSD在初情期从江香猪附睾中的特异性表达模式

3β-羟基甾脱氢酶（3β-HSD）特异性表达于从江香猪的附睾组织中，并且在初情期时表达量最高。而目前对3β-HSD在初情期附睾不同区段的研究未被报道，因此本实验以香猪不同区段的附睾组织作为实验对象，通过使用免疫组织化学（IHC）与蛋白质免疫印迹（WB）分析3β-HSD在初情期香猪附睾5个区段（Ⅰ-Ⅴ区）的具体表达位置和表达量。IHC结果显示3β-HSD定位于香猪附睾的5个区段，且主要定位于附睾管上皮周围的平滑肌、管腔内附睾液及管腔微绒毛中。WB结果显示3β-HSD在Ⅳ区的表达量最高，并且显著高于Ⅰ区、Ⅱ区、Ⅲ区及Ⅴ区。此外，Ⅲ区的表达量显著高于Ⅰ区，但3β-HSD在Ⅰ区、Ⅱ区和Ⅴ区的各组之间并无统计学差异。综上，本研究发现3β-HSD在初情期香猪附睾定位于附睾管管周平滑肌、管腔内附睾液及管腔微绒毛，表达量以Ⅳ区最高，并且以区段特异性表达。     

内皮素受体B在不同毛色羊驼皮肤中的表达与定位

研究内皮素受体B(endothelin recepter B,EDNRB)在羊驼皮肤中的表达与定位,以及在不同毛色中的差异比较,探讨其在羊驼毛色形成中的作用及其相关机制。以不同毛色的成年羊驼为研究对象,用RT-PCR法检测EDNRB基因在不同毛色皮肤中的表达,并使用实时荧光定量PCR技术分析不同毛色羊驼皮肤EDNRB基因的相对表达量,采用Western blot法和免疫组织化学法对EDNRB在羊驼皮肤中的表达进行定位和半定量分析。荧光定量PCR结果显示棕色羊驼中EDNRB基因的相对表达量是白色羊驼的2.1651倍;Western blot结果显示EDNRB蛋白在棕色组的表达显著高于白毛组(P0.05);免疫组化试验结果显示,EDNRB在羊驼皮肤的毛乳头、毛根鞘及表皮细胞中均有表达,且在棕色皮肤组织中显著表达(P0.05)。试验结果提示,EDNRB与羊驼黑色素细胞的活动密切相关,并参与了羊驼毛色和肤色的形成。     

蛋白酶激活受体-2基因mRNA在不同被毛颜色羊驼皮肤组织中的转录分析

蛋白酶激活受体-2(Protease-activated receptor-2,PAR-2)是表达于角质化细胞的膜受体蛋白之一,经证实参与黑色素颗粒的胞间转运,本文通过分析PAR-2基因在不同毛色羊驼皮肤组织中的mRNA转录水平,以期从色素颗粒转运角度研究羊驼不同颜色被毛形成机制。以4头成年白色羊驼和4头棕色羊驼为研究对象,应用实时荧光定量PCR技术,对PAR-2在白色和棕色羊驼皮肤中的mRNA表达水平进行研究。结果显示,PAR-2基因mRNA在棕色羊驼皮肤组织中的相对基因表达量是白色羊驼的2.43倍。结果提示,PAR-2基因mRNA在白色和棕色羊驼皮肤组织中的表达差异表明:毛囊黑色素细胞与角质形成细胞间色素颗粒转运水平是羊驼不同被毛颜色形成的机制之一。     

β-catenin在不同毛色羊驼皮肤中的表达和定位

旨在研究β-catenin在不同毛色羊驼皮肤中的表达和定位,探索其与毛色的关系。以成年白色和棕色羊驼为研究对象,采用实时荧光定量PCR技术、Western blot和免疫组织化学技术,对β-catenin在白色和棕色羊驼皮肤中mRNA、蛋白表达水平和定位进行研究。实时荧光定量PCR结果显示,β-catenin在棕色羊驼皮肤组织中的相对基因表达量是白色羊驼皮肤组织的1.662倍;Western blot结果显示,羊驼皮肤组织粗蛋白提取物中存在分子量约85 ku与兔抗β-catenin多克隆抗体发生免疫阳性反应的蛋白条带,棕色羊驼平均蛋白表达量显著高于白色羊驼;免疫组织化学结果显示,β-catenin在羊驼皮肤的表皮、毛乳头、毛根鞘和皮脂腺中表达,根据光密度值得出,除皮脂腺之外,在表皮、毛乳头和毛根鞘的表达差异极显著(P0.01)。结果提示β-catenin在白色和棕色羊驼皮肤组织中定位以及表达的差异,表明β-catenin参与毛色形成。     

羊驼皮肤中可溶性鸟苷酸环化酶的表达差异

采用免疫组织化学、实时荧光定量PCR和Western blotting对可溶性鸟苷酸环化酶(sGC)在棕色和白色羊驼皮肤中的表达差异进行了研究,以探讨sGC在羊驼皮肤中的作用。实时荧光定量PCR结果显示,sGC在棕色羊驼皮肤中的相对表达量较低,是其在白色羊驼皮肤中的表达量的0.16倍,且差异极显著(P<0.01);Western blot-ting结果显示,在羊驼皮肤总蛋白中含有与兔多克隆抗体发生免疫阳性反应的条带,在棕色皮肤中的反应阳性弱于白色皮肤,且差异极显著(P<0.01);免疫组织化学结果表明,sGC免疫阳性主要分布在白色羊驼皮肤毛囊的毛球上方细胞及细胞间质,而在棕色羊驼皮肤毛囊中,sGC免疫阳性主要分布在毛球底部和外根鞘细胞及细胞间质。结果提示sGC参与羊驼毛色形成。     

Supplementation of graded levels of organic zinc in the diets of female broilers: effects on performance and carcase quality

Zinc is an essential trace element. The objective of this research was to investigate the effects of various levels of organic zinc (OZ) supplementation on growth performance and carcase quality of female broiler chickens. A total of 3200 1-d-old female broiler chicks were randomly allotted to 16 floor pens with 200 birds per pen. A maize-wheat-soyabean meal basal diet (Control) was formulated and 20 mg/kg OZ (20 OZ), 40 mg/kg OZ (40 OZ), and 80 mg/kg OZ (80 OZ) were added to the basal diet to form 4 dietary treatments with 4 replicates per treatment. The OZ source was zinc proteinate which contained 15% zinc. Results showed no significant difference between the treatments in growth performance. A significant increase in thigh skin epidermis and dermis thickness was shown in the OZ supplementation groups; however, no effect was found on the thickness of back skin epidermis and dermis. Collagen contents in breast and thigh meats were not influenced by OZ supplementation but a significant increase in collagen content was found in the back and thigh skin. This increase in collagen content was significantly greater in the back and thigh skin of OZ 80 than with OZ 20. Shear force value and zinc concentration in skins and meat were not significantly influenced by supplementation with OZ. It is concluded that dietary OZ does not improve growth performance of broilers; however, it could increase skin thickness by increasing collagen content in skin, thereby improving carcase quality.     

梅花鹿颈背部异常松弛皮肤光镜组织学观察

本研究利用Weigert’s-Van Giesson染色法和Masson氏三色染色法分别对正常梅花鹿和颈背部皮肤异常松弛的梅花鹿皮肤中的弹性纤维和胶原纤维进行观察研究,探讨梅花鹿松弛皮肤组织学致病特点。结果表明,正常梅花鹿真皮网状层中有较多的弹性纤维有规律地平行排列,呈波浪状,染色均匀;真皮乳头层有少量弹性纤维垂直于表皮分布。皮肤松弛梅花鹿或是真皮乳头层中缺乏弹性纤维（大0号和79号鹿）,或是网状真皮层中弹性纤维严重发育不全,出现集聚、断裂现象（33号和79号）,且排列紊乱、呈不规则状（大0号）;正常梅花鹿皮肤胶原纤维在真皮层中呈多层规律性分布。皮肤松弛梅花鹿胶原纤维束比正常胶原束粗大、胶原纤维束减少、堆积、排列杂乱无序,胶原纤维间表现为严重的稀疏。这些现象与人类皮肤松弛症的皮肤组织学症状非常相似。     

Review: The Role of Collagen in Meat Tenderness

Meat tenderness is a major factor affecting the consumers’ assessment of meat quality. Collagen is an abundant connective tissue protein and is a contributing factor to variation in meat tenderness and texture. It is found in three specific regions in muscle and in various forms and types. Collagen molecules are bound together through intermolecular crosslinks that help provide structure and strength. These crosslinks are initially reducible, but over time are replaced by mature, thermally stable, and less soluble crosslinks. These mature crosslinks, rather than the total amount of collagen, are the key factors in collagen-related toughness. The proportion of mature to reducible crosslinks increases with age, resulting in older animals that often have less tender meat than younger animals. Collagen crosslinks are also affected by growth rate, nutrition, and genetics. Collagen plays major roles in cooked meat as well. As collagen fibrils are heated during cooking, they shrink, resulting in a fluid loss and less tender meat. They also act to hold muscle fibers together after shrinkage. Post-mortem degradation of collagen and the use of collagenases appear to play a role in providing the desired tenderness and texture by altering the connective tissue structure. Collagen is very important in maintaining acceptable texture; however, high amounts of crosslinks can greatly decrease tenderness and consumer acceptability.     

胶原蛋白交联的研究进展及其对肉的纹理和嫩度的影响

胶原蛋白是肌间结缔组织的主要成分 ,它在肌肉中的含量和质量对肌肉纹理性状的影响实际上是取决于胶原蛋白分子间交联的水平、方式和成熟程度。影响胶原蛋白分子间交联的因素很多 ,包括动物的品种、年龄、性别和肌肉的来源以及其它一些与生长有关的环境因素。本文着重对由一系列酶介导的胶原蛋白分子间的共价交联、胶原蛋白在细胞外的被动修饰、胶原蛋白对烤制后肌肉纹理的影响、胶原蛋白分子间交联及其影响因素和调控机制进行综述和探讨     

Practical treatment of body and open leg wounds of horses with bovine collagen,biosynthetic wound dressing and cyanoacrylate

Horses with severe and deep lacerations are represented by ten cases in which treatment emphasized the use of bovine collagen preparations to promote controlled second-degree repair. Traumatized areas were tarsal, metatarsal, neck, forearm, metacarpal and pastern. Wound size changes were recorded. Depending on the wound type, the site was treated with antibiotic-steroid ointment, organic acid cream, sterile collagen particles, suspension or dressings protected by hydrogel dressing, non-adherent pads, occlusive skin dressings, roll gauze and elastic tape. In three cases, a fiberglas cast was applied over a hind leg wound and lacerated tendon for stability. When controlled granulation of the deeper wounds reached skin level, the area often was stabilized by only cyanoacrylate spray. As these cases presented a wide range of trauma each with a unique history, healing rates were based on initial measurements. An overall progression of wound reduction occurred at a predictable rate. The exogenous collagen formulations were used to stimulate controlled granulation, ie. to “jump start” the healing process. Collagen particles, suspension or dressings were packed into depressions, placed under suture lines, secured over abraded tissue, and placed under protective bandage or cast. To further evaluate the use of cyanoacrylate tissue spray in wound treatments, an additional ten cases are presented. The variety of wounds were produced experimentally in Center-owned ponies or provided as clinical cases. Wound size changes and healing progress were recorded. Wounds occurred on the neck, abdomen, metacarpal, metatarsal, fetlock and pastern areas. Depending on wound type, the site was treated with cyanoacrylate only; or treated as above until controlled granulation attained skin level. In one case punch grafts of skin were transferred from one foreleg to the opposite with the horse standing. Cyanoacrylate spray provided a water proof barrier protecting the wound from dirt, debris and insects as well as stabilizing full-thickness skin lacerations by bridging normal to traumatized skin allowing uninterrupted granulation and epithelialization. The use of a neck cradle prevented wound disturbance and stall confinement aided stabilization.     

Reduced anchoring fibril formation and collagen VII immunoreactivity in feline dystrophic epidermolysis bullosa

Dystrophic epidermolysis bullosa was diagnosed in a cat with juvenile-onset epithelial sloughing of the oral mucosa, footpads, and haired skin. Dermoepidermal separation occurred in the absence of inflammation or cytolysis of basal epidermal cells. Collagen IV-specific immunostaining corroborated the fact that clefting took place below the epidermal basement membrane. Ultrastructural examination revealed that the proband's anchoring fibrils exhibited a filamentous morphology and were decreased in number compared with those in a normal cat. Finally, the attenuated immunoreactivity for collagen VII in our patient led us to suspect that its encoding gene, COL7A1, could be mutated in this case of feline dystrophic epidermolysis bullosa.     

Factors regulating collagen synthesis and degradation during second-intention healing of wounds in the thoracic region and the distal aspect of the forelimb of horses

OBJECTIVE: To determine significant molecular and cellular factors responsible for differences in second-intention healing in thoracic and metacarpal wounds of horses. ANIMALS: 6 adult mixed-breed horses. PROCEDURE: A full-thickness skin wound on the metacarpus and another such wound on the pectoral region were created, photographed, and measured, and tissue was harvested from these sites weekly for 4 weeks. Gene expression of type-I collagen, transforming growth factor (TGF)-beta1, matrix metalloproteinase (MMP)-1, and tissue inhibitor of metalloproteinase (TIMP)-1 were determined by quantitative in situ hybridization. Myofibroblasts were detected by immunohistochemical labeling with alpha-smooth muscle actin (alpha-SMA). Collagen accumulation was detected by use of picrosirius red staining. Tissue morphology was examined by use of H&E staining. RESULTS: Unlike thoracic wounds, forelimb wounds enlarged during the first 2 weeks. Myofibroblasts, detected by week 1, remained abundant with superior organization in thoracic wounds. Type-I collagen mRNA accumulated progressively in both wounds. More type-I collagen and TGF-beta1 mRNA were seen in forelimb wounds. Volume of MMP-1 mRNA decreased from day 0 in both wounds. By week 3, TIMP-1 mRNA concentration was greater in thoracic wounds. CONCLUSIONS AND CLINICAL RELEVANCE: Greater collagen synthesis in metacarpal than thoracic wounds was documented by increased concentrations of myofibroblasts, type-I collagen mRNA,TGF-beta1 mRNA, and decreased collagen degradation (ie, MMP-1). Imbalanced collagen synthesis and degradation likely correlate with development of exuberant granulation tissue, delaying healing in wounds of the distal portions of the limbs. Factors that inhibit collagen synthesis or stimulate collagenase may provide treatment options for horses with exuberant granulation tissue.     

鹿产品中胶原蛋白的研究及应用进展

胶原蛋白作为一种重要的功能性蛋白质已被广泛应用于食品、化妆品、生物及医学等各个领域。鹿产品,作为一种新型胶原蛋白资源,对其进行研究与开发具有重要意义。文中就鹿产品胶原蛋白的研究及应用现状进行概括,为鹿产品胶原蛋白的深度开发利用提供参考。     

The comparison of the collagen fiber contents and hepatic stellate cell distribution in male and female chicken livers

The difference in collagen fiber content, morphological properties and hepatic stellate cell (HSC) distribution was investigated in the liver of both sexes in chicken. Collagen fiber specimens were obtained by maceration treatment with NaOH solution. HSCs were detected using desmin‐specific immunohistochemistry. The ratio of liver weight to body weight was larger in the female than the male chickens. Collagen fiber content, the numerical density of HSCs and the percentage area displaying desmin immunopositivity were not different between the right and left lobes of the liver, in both male and female chickens. However, all of these parameters were larger in the males than the females. In the light microscopic observation, many HSCs in the male had large and elongated cytoplasmic processes. Conversely, HSCs with poorly developed cytoplasmic processes were frequently observed in females. Liver tissue is structurally stronger in male chickens than females and the activity and density of HSCs may be related to the collagen fiber content in chicken liver.