野生型小鼠皮肤cyclinA1 mRNA的表达及其意义

目的从RNA水平探讨cyclinA1在野生型小鼠皮肤中的表达及意义。方法选取30只大于6个月的野生型小鼠,用原位杂交方法定位检测cyclinA1 mRNA在头颈部皮肤中的表达,同时以不加探针皮肤标本作为阴性对照,另取雄性小鼠睾丸组织作为阳性对照。结果分别有25只野生型小鼠在皮脂腺部位及12只在表皮全层有cyclinA1 mRNA表达。其中,皮脂腺部位强阳性及阳性表达分别占50.0%和33.3%,阳性率为83.3%;表皮全层强阳性及阳性表达分别占13.3%和26.7%,阳性率为40.0%。皮脂腺阳性率显著高于表皮(P0.05)。结论CyclinA1 mRNA在野生型小鼠头颈部皮肤的皮脂腺部位及表皮全层的表达均有较高的阳性率,尤其皮脂腺表达的阳性率更高。     

Mdr2基因敲除小鼠建立原发性肝癌模型观察

目的 观察C57BL/6背景的Mdr2基因敲除小鼠自发肝肿瘤形成情况。方法 (11.3±4.2)周龄Mdr2基因敲除C57BL/6-Abcb4^tm1小鼠9只和野生型C57BL/6小鼠5只,连续饲养65周后处死小鼠,留取血清及肝标本。检测血清ALT、AST、AFP水平,肝组织石蜡切片做HE、天狼猩红染色,免疫组织化学检测肿瘤及肿瘤旁组织CK-7、CK-19表达情况。结果 9只Mdr2基因敲除小鼠均自发形成肝肿瘤,血清ALT、AST、AFP水平均显著高于野生型小鼠(P<0.01),Mdr2基因敲除小鼠肝肿瘤CK-7、CK-19染色均为阴性。结论 Mdr2基因敲除小鼠连续饲养至(76.3±4.2)周龄时均自发形成肝肿瘤,其病理组织分型为肝细胞癌。     

一种皮肤型狼疮小鼠模型的鉴定

目的通过对老龄系统性红斑狼疮小鼠(TC)皮肤症状的观察和鉴定,探讨其是否可以作为一种研究皮肤型狼疮的小鼠模型。方法观察TC小鼠出现皮肤症状的年龄,对比同龄小鼠与C57BL/6小鼠的皮肤症状,ELISA法检测小鼠血清抗dsDNA抗体及抗ANA抗体,HE染色检测小鼠皮肤病理学症状。结果 2/3的TC小鼠出现毛发脱失、皮肤溃烂等症状,其病变最早发生于40周龄,皮肤病变的狼疮小鼠血清抗dsDNA抗体和抗ANA抗体滴度明显高于C57BL/6小鼠(P<0.05);HE染色结果显示狼疮小鼠角质层缺失,上皮层连续性中断,真皮层大量淋巴细胞浸润。结论狼疮小鼠在40周后会出现皮肤病变,显示该小鼠可作为一种研究慢性皮肤型狼疮的疾病模型。     

脱细胞羊膜与小肠黏膜下层促进大鼠皮肤缺损修复和血管形成

目的观察脱细胞羊膜(HAM)与小肠黏膜下层(SIS)促进大鼠皮肤缺损修复和血管形成的作用。方法SD大鼠24只,在两侧背部各做1个直径为1.8cm圆形全层皮肤缺损。创面随机分为A组、B组和C组。A组HAM覆盖,B组SIS覆盖,C组纱布覆盖。在2周时处死动物取材,HE染色观察皮肤缺损修复情况。免疫组织化学染色检测K19和VEGF,RT-PCR检测VEGF mRNA的表达。结果A组、B组愈合较好。C组愈合较差。免疫组织化学染色显示,A、B组K19、VEGF阳性细胞显著多于C组,其中A组最多;RT-PCR结果显示,A、B组比C组表达更多的VEGF mRNA其中A组最多,差异显著,有统计学意义(P<0.05)。结论HAM和SIS均能增加皮肤创面组织中K19阳性细胞数,上调VEGF mRNA的表达、增加VEGF的分泌,其中HAM具有更强的促进皮肤缺损修复和血管形成作用。     

链脲佐菌素糖尿病小鼠睾丸内生精细胞减少与PCNA和Ki67表达降低

目的通过检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及Ki67在正常及糖尿病小鼠睾丸组织中表达的差异,探讨糖尿病对生精细胞增殖的影响。方法 30只正常雄性C57BL/6小鼠,随机分为对照组(10只)和糖尿病组(20只)。采用腹腔注射链脲佐菌素的方法建立糖尿病小鼠模型,对照组注射相同体积的柠檬酸—柠檬酸钠缓冲液。造模成功3周后处死动物,取睾丸组织,常规固定、石蜡包埋、切片HE染色观察睾丸组织的形态学变化;免疫组织化学SP法检测PCNA及Ki67在睾丸组织中的表达;图像分析技术分析组织中PCNA及Ki67免疫组织化学表达水平。结果 HE染色显示,糖尿病小鼠睾丸内处于精子发生前半期的生精小管内生精细胞排列疏松,细胞层数偏少,附睾管内精子密度相对较低;免疫组织化学染色显示,PCNA和Ki67在糖尿病小鼠睾丸生精小管中的表达均明显降低。结论高糖可能通过降低生精细胞的增殖进而影响睾丸精子的发生。     

选择性敲除小鼠神经细胞adam10基因导致胼胝体和海马联合发育不全

目的探讨adam 10基因在小鼠神经系统发育的作用。方法将gfapcre转基因鼠和adam10loxlox转基因鼠杂交,得到神经细胞特异性adam10基因敲除鼠(gfapcre-adam10loxlox);在出生后第14d对小鼠大脑切片行HE染色和免疫组织化学染色,观察adam10基因敲除后神经系统发育情况。结果选择性敲除小鼠神经细胞adam10基因(gfapcre-adam10loxlox)后,小鼠可存活至出生后3w左右;HE染色发现胼胝体缺失,海马发育不全,髓鞘碱性蛋白(myelin basic protein,MBP)免疫荧光染色发现髓鞘发育异常。结论 Adam10基因在小鼠神经系统发育中具有重要作用,选择性敲除小鼠神经细胞adam10基因导致胼胝体和海马联合发育不全,髓鞘发育异常。     

脱钙对鼠颅骨标本中肥大细胞组织化学与免疫组织化学染色的影响

目的:探讨脱钙对鼠颅骨标本中肥大细胞组织化学与免疫组织化学染色影响.方法:用不同脱钙液处理小鼠颅骨标本,组织切片后进行苏木素一伊红染色、甲苯胺蓝染色、醛品红染色以及免疫组织化学染色.结果:经过不同脱钙液处理后的颅骨组织切片组织结构保存完好,肥大细胞的组织化学染色(甲苯胺蓝和醛品红染色),及肥大细胞中胰蛋白酶的免疫组织化学染色清晰.结论:不同脱钙液(8%盐酸脱钙液、EDTA脱钙液、混合酸脱钙液)处理不影响鼻腔粘膜中肥大细胞的组织化学和免疫组织化学染色.     

Notch 2在肌萎缩侧索硬化症SOD1~(G93A)转基因小鼠脊髓内的表达变化

目的明确肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)SOD1G93A转基因小鼠脊髓内Notch 2表达水平与ALS病程的关系。方法应用Notch基因阵列表达谱检测Notch信号通路相关分子在表达人突变SOD1基因的SOD1G93A转基因ALS小鼠及野生型鼠脊髓内的表达差异。应用real-time PCR、免疫组织化学染色、Western blot方法检测Notch 2在不同阶段各组小鼠脊髓内的表达变化。结果 Notch基因阵列表达谱聚类分析结果显示Notch 1、Notch 2在95d、122d SOD1G93A鼠脊髓内均显著增高,而Notch 4仅在122d SOD1G93A鼠升高明显。Notch下游靶基因HES1在108d SOD1G93A鼠脊髓内显著降低。RT-PCR结果证实Notch 2在SOD1G93A鼠脊髓内的表达变化。免疫组织化学标记显示Notch 2在SOD1G93A鼠、野生型鼠脊髓各部均有表达,Western blot结果显示Notch 2蛋白表达在95d SOD1G93A鼠增高显著。结论 Notch 2信号分子的表达随ALS疾病进展,在脊髓退变过程中变化显著,可能参与ALS疾病进程。     

内质网应激介导小鼠急性汞中毒性肝损伤

目的 观察急性汞中毒对小鼠肝的损伤,并探讨内质网应激是否介导此损伤.方法 选取8周龄健康雄性C57B/L6小鼠,随机分为对照组、HgCl2染毒组、4-PBA预处理+HgCl2染毒组,建模成功后,眼球取血检测小鼠肝功能变化;HE染色观察肝形态学变化;免疫组织化学染色及Western blot检测肝组织中内质网应激特异性蛋...     

AQP1在CD-1纯系野生型孕鼠胎盘组织的表达

目的：研究水通道蛋白1（Aquaporin 1,AQP1）在小鼠胎盘组织的分布及表达,初步探讨AQP1在羊水循环及母胎液体平衡中的作用。方法：各取四只雌雄成年健康野生型CD1小鼠（wild type,AQP1＋/＋）及AQP1基因敲除小鼠（AQP1-KO,AQP1-/）-,将纯合子AQP1基因敲除雌雄小鼠等数量合笼交配,第二日检出阴道栓者记为妊娠第1天（1 gestational day,1GD）;野生型小鼠同样合笼记录。分别取两组13GD孕鼠的胎盘组织各一个,应用逆转录-聚合酶链反应（RT-PCR）技术及免疫组织化学技术检测AQP1胎盘组织中的表达,并确定AQP1在小鼠胎盘组织的定位。结果：1.RT-PCR结果表明AQP1在CD-1野生型孕鼠胎盘组织表达,AQP1基因敲除鼠无表达;2.免疫组织化学方法发现AQP1表达于小鼠胎盘血管内皮细胞和滋养细胞,AQP1基因敲除鼠无表达。结论：在mRNA水平和蛋白水平均发现AQP1在CD-1纯系野生型孕鼠胎盘组织的表达,提示AQP1可能在羊水循环及母胎液体平衡中发挥作用。     

The Clouston syndrome mutation connexin30 A88V leads to hyperproliferation of sebaceous glands and hearing impairments in mice

Distinct mutations in the gap junction protein connexin30 (Cx30) can cause the ectodermal dysplasia Clouston syndrome in humans. We have generated a new mouse line expressing the Clouston syndrome mutation Cx30A88V under the control of the endogenous Cx30 promoter. Our results show that the mutated Cx30A88V protein is incorporated in gap junctional plaques of the epidermis. Homozygous Cx30A88V mice reveal hyperproliferative and enlarged sebaceous glands as well as a mild palmoplantar hyperkeratosis. Additionally, homozygous mutant mice show an altered hearing profile compared to control mice. We conclude that the Cx30A88V mutation triggers hyperproliferation in the skin and changes the cochlear homeostasis in mice.     

Ragged--a new rexoid mutant rat with large sebaceous gland

A new hair defected mutant rat was established. This mutant was covered with ragged hair since about 10 days of age, then transiently lost most of hair in the back at approximately 5 weeks of age and re-covered with ragged hair thereafter. Thickened eyerids occurred since about 3 weeks of age. Histological examination revealed enlarged sebaceous glands with greater number of sebaceous cells in the back skin. The oil stained skin samples showed normal sebaceous transformation and pilosebaceous canal. Genetical analysis showed that the ragged hair character was a single recessive trait and indicated that this single recessive gene was not linked with the coat color genes, non-agouti (a), albino (c) and hooded (h). From the present data and previous reports, we recommended this single recessive gene is a new rexoid mutation thereby we termed this gene "Ragged (rg)".     

Phospholipase Cdelta1 is required for skin stem cell lineage commitment

Phosphoinositide-specific phospholipase C (PLC) is a key enzyme in phosphoinositide turnover and is involved in a variety of physiological functions. Here we report that PLCdelta(1)-deficient mice undergo progressive hair loss in the first postnatal hair cycle. Epidermal hyperplasia was observed, and many hairs in the skin of PLCdelta(1)-deficient mice failed to penetrate the epidermis and became zigzagged owing to occlusion of the hair canal. Two major downstream signals of PLC, calcium elevation and protein kinase C activation, were impaired in the keratinocytes and skin of PLCdelta(1)-deficient mice. In addition, many cysts that had remarkable similarities to interfollicular epidermis, as well as hyperplasia of sebaceous glands, were observed. Furthermore, PLCdelta(1)-deficient mice developed spontaneous skin tumors that had characteristics of both interfollicular epidermis and sebaceous glands. From these results, we conclude that PLCdelta(1) is required for skin stem cell lineage commitment.     

Histological characteristics of the pelage skin of rough fur mice (C3H/HeJ- ruf/ruf).

Pelage skin of C3H/HeJ mice homozygous at an autosomal recessive mutant locus, rough fur (ruf) which is located on chromosome 9, was histologically analyzed. Sebaceous glands synthesizing lipids were larger in the mutant mice than in controls in an examination by Sudan IV staining. Electron microscopic analysis of the sebaceous gland showed that lipid droplets were denser in mutant mice than in control mice, and that they were irregular in shape in ruf mice while those of controls were round. Our results suggested that rough fur (ruf) mice might be an animal model for hyperlipogenesis of the pelage skin.     

Intraductal papilloma of the salivary gland. A report of two cases with diagnosis by fine needle aspiration biopsy

BACKGROUND: Intraductal papillomas are rare, benign tumors most commonly encountered in minor salivary glands. They are cystic, solitary neoplasms that arise from ductal epithelium and produce painless swellings. CASES: Two cases arose in major salivary glands. The first case was a superficial, firm mass at the superior edge of the parotid, cytologically evocative of an adnexal tumor. A firm, submandibular mass in the second case was diagnosed as a papillary neoplasm. Fluid was aspirated from both cases. Three-dimensional epithelial clusters, some with a papillary configuration and histiocytes, were the main cellular components. The majority of cells showed oncocytic differentiation; however, benign-appearing ductal cells in honeycomb sheets were also present. The first case also had occasional cells suggestive of sebaceous differentiation. The excised lesions were unilocular cystic papillary neoplasms consistent with intraductal papilloma; focal sebaceous differentiation was noted in the first case. CONCLUSION: Awareness of the cytologic features of intraductal papilloma of the salivary glands should prompt its inclusion in the differential diagnosis of papillary lesions of the head and neck.     

Inherited seborrheic dermatitis--a new mutant in mice

A new mutation, affecting skin and hair, occurred in an expansion colony of Him:OF1 mice. Test crosses showed that a single autosomal recessive gene was responsible for this trait. Homozygotes have sparse greasy fur and lower viability and fertility than normal littermates. Histological observations showed hypertrophy of sebaceous glands, hyperkeratosis, parakeratosis, acanthosis and signs of inflammation. The disease was named 'inherited seborrheic dermatitis' and the gene name seb is proposed.     

Characterization of Hair Follicle Development in Engineered Skin Substitutes

Generation of skin appendages in engineered skin substitutes has been limited by lack of trichogenic potency in cultured postnatal cells. To investigate the feasibility and the limitation of hair regeneration, engineered skin substitutes were prepared with chimeric populations of cultured human keratinocytes from neonatal foreskins and cultured murine dermal papilla cells from adult GFP transgenic mice and grafted orthotopically to full-thickness wounds on athymic mice. Non-cultured dissociated neonatal murine-only skin cells, or cultured human-only skin keratinocytes and fibroblasts without dermal papilla cells served as positive and negative controls respectively. In this study, neonatal murine-only skin substitutes formed external hairs and sebaceous glands, chimeric skin substitutes formed pigmented hairs without sebaceous glands, and human-only skin substitutes formed no follicles or glands. Although chimeric hair cannot erupt readily, removal of upper skin layer exposed keratinized hair shafts at the skin surface. Development of incomplete pilosebaceous units in chimeric hair corresponded with upregulation of hair-related genes, LEF1 and WNT10B, and downregulation of a marker of sebaceous glands, Steroyl-CoA desaturase. Transepidermal water loss was normal in all conditions. This study demonstrated that while sebaceous glands may be involved in hair eruption, they are not required for hair development in engineered skin substitutes.     

Rac1 is crucial for hair follicle integrity but is not essential for maintenance of the epidermis

Rac1 is a small GTPase that regulates the actin cytoskeleton but also other cellular processes. To investigate the function of Rac1 in skin, we generated mice with a keratinocyte-restricted deletion of the rac1 gene. Rac1-deficient mice lost nearly all of their hair within a few weeks after birth. The nonpermanent part of mutant hair follicles developed constrictions; lost expression of hair follicle-specific keratins, E-cadherin, and alpha6 integrin; and was eventually removed by macrophages. The permanent part of hair follicles and the sebaceous glands were maintained, but no regrowth of full-length hair follicles was observed. In the skin of mutant mice, epidermal keratinocytes showed normal differentiation, proliferation, cell-cell contacts, and basement membrane deposition, demonstrating no obvious defects of Rac1-deficient epidermis in vivo. In vitro, Rac1-null keratinocytes displayed a strong spreading defect and slightly impaired adhesion. These data show that Rac1 plays an important role in sustaining the integrity of the lower part of hair follicles but not in maintenance of the epidermis.     

Transient expression of wild type and mutant human apolipoprotein AI in COS cells

A human apolipoprotein AI (apo AI) minigene and two mutants were cloned into the vector pUHD10-1 for expression studies in COS cells under the control of the strong CMV (cytomegalovirus) enhancer and the own apo AI promoter. In the mutated apo AI minigene (mutant M1) the positions of the triplets of Gln(-2)-Gln-1 at the C-terminus of the prosequence were exchanged against Gln(-8)-Ala-7, the recognition site of the signal peptidase of the wild type human apo AI. The prosequence has been deleted in mutant M2 and the presequence linked directly to the N-terminus of the mature apo AI form. We report here on expression studies in COS cells, a cell line, which does not express apo AI. They were transfected by electroporation with pUHD10-1 constructs, which contain a) the wild type apo AI minigene and b) the two mutant apo AI minigenes with mutations described above. The following results were obtained: a) the wild type and mutant apo AI constructs were efficiently transcribed and translated in COS cells, b) the expression of the wild type preproapo AI minigene in COS cells led to the secretion of proapo AI (29 kDa), that of the mutant (M2) gene, devoid of the prosequence of mature apo AI (28.4 kDa), whereas the product of mutant gene M1 (31 kDa) with the recognition site of the signal peptides transposed to the C-terminus of the prosequence remained uncleaved within the COS cells.(ABSTRACT TRUNCATED AT 250 WORDS)