山羊毛囊干细胞的分离及体外培养

试验采用2．4U／mL Dispase酶消化和机械切割法分离毛囊隆突部,经胰酶（0．5mg／mL胰酶＋0．2mg／mL EDTA）消化从山羊耳部皮肤分离得到的毛囊干细胞,然后进行形态学观察、细胞生长曲线测定、克隆形成率及免疫组化染色检测,并对毛囊干细胞的基础培养基进行了筛选。结果表明：DMEM／F12是一种适合毛囊干细胞体外增殖培养的培养基;在以DMEM／F12为基础培养液的培养体系中细胞可传代至19代。     

羊驼毛囊干细胞分离、培养与鉴定

采用0．2％中性蛋白酶Ⅱ和0．25％胰酶：0．02％EDTA（1：1）“两步酶”消化法从羊驼背部皮肤分离得到羊驼毛囊干细胞。用无血清角质细胞培养基（K-SFM）培养进行形态学观察、细胞生长曲线克隆形成率及免疫组化染色检测。结果显示,细胞生长曲线表明,不同代次毛囊干细胞接种前3d生长缓慢,4～6d进入倍增期,有无限增殖的趋势,所分离的羊驼毛囊干细胞具备毛囊干细胞特征（体积小、立体感强）,呈现未分化特征;CK19、CK15、81-integrin和CD34免疫组化染色阳性;第3、5、7、9代克隆形成率分别为（32．7±2．27）％、（47．0±3．46）％、（46．3±3．18）％和（43．3±3．76）％。结果表明,用“两步酶”消化法成功分离获得羊驼毛囊干细胞,无血清角质细胞培养基（K—SFM）可使羊驼毛囊干细胞体外维持未分化状态并传代至12代。     

毛囊干细胞研究进展

文章阐述了毛囊干细胞的定位、表面标记及分化调控,同时阐述了毛囊干细胞在毛发再生、创面愈合、决定动物皮毛颜色基因程序重调及其它方面的应用。对实现毛发再生,促进皮肤损伤后功能与结构的完全修复具有重要意义。     

Immunohistochemical detection of p27 and p21 proteins in canine hair follicle and epidermal neoplasms

Trichoblastomas, trichoepitheliomas, and squamous cell carcinomas in the skin of dogs were analysed by immunohistochemistry for the nuclear expression of p27, p21 and proliferating cell nuclear antigen (PCNA). High levels of p27 were present in trichoepitheliomas and trichoblastomas compared with squamous cell carcinomas. Detectable p21 was found in trichoepitheliomas and squamous cell carcinomas, but trichoblastomas had low level of p21 nuclear reactivity. Low levels of PCNA were detected in trichoepitheliomas and trichoblastomas compared with squamous cell carcinomas. The results suggested that nuclear p27 acts as a cyclin-dependent kinase (CDK) inhibitor in trichoepitheliomas and trichoblastomas. Nuclear p21 expression is involved in the induction of epithelial differentiation and seems to be unrelated to CDK inhibition.     

The effects of thyroxine on hair growth in the dog

The effects of orally and topically administered thyroxine on hair growth were studied in normal, intact, cross-bred male dogs maintained under the same controlled conditions of temperature and photoperiod. It was found that, irrespective of the route of administration, thyroxine produced an increase in both the rate of hair growth and in the number of hair follicles entering the anagen phase of the hair cycle. With orally administered thyroxine, the changes in hair growth were most marked in the flank region.     

Primary and secondary bone tumours in the dog

During a 14-year period (January 1962 to December 1975), 394 primary and secondary tumours or tumour-like lesions affecting bones were diagnosed in the dog. The most frequently occurring tumour was osteosarcoma (183, 46.5%), followed by chondrosarcoma (forty-four, 11.2%), squamous cell carcinoma (thirty-nine, 10%), fibrosarcoma (thirty-one, 7.8%), multiple myeloma (fourteen, 3.6%), haemangiosarcoma (fourteen, 3.6%), malignant melanoma (thirteen, 3.3%), synovioma (nine, 2.3%), lymphosarcoma (eight, 2.0%), cartilage analogue of fibromatosis (seven, 1.8%), adaman-tinoma (five, 1.3%), osteocartilaginous exostosis, ossifying fibroma, rhabdomyosarcoma (four each), polyostotic cyst, haemangiomatosis (three each), odontoma, tumoral calcinosis (two each), and osteoma, chondroma, desmoid fibromatosis, and malignant mesenchymoma (one each).
Of these, desmoid fibromatosis, malignant mesenchymoma, and haemangiomatosis have not been previously reported in the dog.     

Nerve sheath tumours in the dog and cat

Schwannomas were diagnosed in twelve dogs and five cats at Massey University Small Animal Clinic and Hospital over a 15-year period (1977-92). A further two feline cases were reported at the Batchelar Animal Health Laboratory. In six dogs, the tumour involved nerves of the brachial plexus. Clinical signs observed in these dogs were forelimb lameness, muscle wasting and pain on movement of the affected limb or neck. Hindlimb paresis was observed in two dogs. Surgical excision of the brachial plexus tumour was attempted in one dog, leading to an 8-month remission of signs. In one dog, the tumour involved the sacral nerves, and in two dogs the cranial nerves were affected. Three dogs had skin nodules. Seven of the twelve affected dogs were destroyed. In five cats, the tumours developed on the carpus, tarsus or interdigital area of a forelimb or hindlimb as a slowly developing nodular lesion. In the other two cats, the site of the tumour was the flank and the lateral thigh respectively. Surgical excision of the tumour was successful in three cats.     

Determination of the depth of excision using a dermatome (Aesculap) to export all hair follicle bulbs from a donor site in the dog

The aim of the study was to determine the depth of excision using a dermatome to excise all hair follicle bulbs from the donor site. Partial-thickness skin samples of different depth, ranging from 0.4 to 1 mm, were excised using a dermatome (Aesculap) from the dorsal aspect of the trunk of the dog. Biopsies, using a 6-mm biopsy punch, were performed in the centre of each donor site and excised sample. They were processed routinely for histological examination and the aspect of the hair follicles was observed. Split-thickness skin grafts of >or=0.7 mm deep performed with a dermatome contain the entire hair follicle.     

The clinical diagnosis and management of rectal and pararectal tumours in the dog

Forty-two dogs bearing tumours of the rectum or the pararectal tissues were examined over a 3 year period. The presenting clinical signs, diagnostic techniques and management are described. The prognosis was found to be guarded irrespective of the histological type. The mean survival times for malignant epithelial tumours of the rectal wall was 6–9 months, for rectal lymphomas 6-5 months and for pararectal tumours 10-6 months. The major limiting factor was found to be local recurrence after surgery for rectal epithelial tumours, resistance to chemotherapy and local recurrence for rectal lymphomas and metastatic disease for pararectal tumours.     

Secondary bone tumours in the dog.

During a 2 1/2-year period 209 dogs with 218 malignant tumours were examined for skeletal metastases. There were 77 malignant tumours without visceral or skeletal metastases. Twenty-four (17%) of the 141 tumours with visceral metastases also had skeletal metastases. Ninety-eight of these tumours were of epithelial origin, and of these 21 (21.4%) had skeletal metastases. In 14 dogs, more than one bone had a metastatic tumour. The humerus, femur and vertebral column were most frequently involved. The lung was the most frequent site of metastatic tumours; then, in order, the liver, kidney and skeleton. The incidence of primary and secondary bone tumours was similar. Metastatic bone disease in the dog is far more common than is generally realized.     

豫医卷毛大鼠皮肤毛囊形态学及超微结构观察

为比较卷毛大鼠与SD大鼠皮肤毛囊形态学区别,探讨卷发机理,采用病理组织学和电镜技术观察突变大鼠皮肤毛囊结构。组织学结果显示卷毛大鼠皮肤毛囊弯曲,毛囊皮质内根鞘鞘小皮层排列不规则。扫描电镜下卷毛大鼠毛干弯曲,毛干横断面为扁平形,根鞘厚薄不均。透射电镜观察发现突变大鼠毛囊内根鞘细胞内见多量透明蛋白颗粒与角蛋白丝形成的均质状复合物,排列不规则。研究结果表明,突变基因可能引起毛囊内根鞘细胞角化过程不一致和中间丝的不规则排列而导致卷发。     

A study of normal hair growth in the dog

The rate of hair growth in the forehead, shoulder and flank regions of four normal-coated, cross-bred, male dogs maintained under the same controlled conditions of photoperiod, ambient temperature and diet, was measured during two six-week periods, one during summer and the other in winter. It was found that the rate of hair growth varied between individual dogs and also between the different regions in the same dog. Growth rate was most rapid in the shoulder region, followed by the flank and then the forehead regions, and was slightly more rapid in the shoulder and flank regions of three of the dogs during summer than in winter. Possible explanations for these variations in hair growth rate are discussed.     

Cytologic features of testicular tumours in dog

In this paper, we report on our experience of cytology of fine needle biopsies performed on 92 dogs with testicular tumours during the period from 1998 to 2002. Cytological diagnosis was consistent with seminoma in 20 cases, sertolioma in 16 cases, Leydig cell tumours in 50 cases and mastocytoma in one case. Five cases could not be diagnosed by cytology. Cytological observations were confirmed after surgery by histopathological examination in 87 cases. Cytology provided a sensitivity of 95% for seminoma, 88% for sertolioma and 96% for Leydig cell tumours. The specificity was 100% for all three tumour types. In our experience cytology of fine needle aspirations of testicular tumours is a very reliable technique.     

The reaction of hair follicles in the vicinity of full-thickness excised skin wounds in the dog

The changes which take place in hair follicles close to full-thickness excised skin wounds in the thoracic and metatarsal regions of the dog are described. The follicles make a significant contribution to the bulk of epithelial cells which are capable of migrating to form the new epidermis, and their behaviour explains why the migrating epithelium is first seen to grow on to the surface of the wound in the sector which lies in the direction of hair flow, and why a zone of alopecia surrounds the excised wound.     

Brain stem auditory-evoked responses in the dog

Brain stem auditory-evoked responses (BAER) were recorded from 58 dogs that did not have a known history of hearing problems. The BAER wave forms had an overall mean amplitude approximately 3.0 microV and typically consisted of a series of 4 to 5 vertex-positive peaks (peaks I through V). When acoustic clicks having intensities of 60-dB hearing level (decibels relative to the subjective hearing threshold) were used as stimuli, peak I had a latency of 1.49 +/- 0.13 ms; peak II, 2.32 +/- 0.14 ms; peak III, 3.01 +/- 0.25 ms; peak IV, 4.22 +/- 0.27 ms; and peak V, 5.55 +/- 0.37 ms. Latency values were influenced by a number of nonpathologic factors, including stimulus intensity and the body temperature of the dog. As stimulus intensity was decreased, there was a lengthening of the latency of each peak coupled with a decrease in the overall amplitude of BAER. Decreases in rectal temperature caused a similar lengthening of peak latencies. Age may have an influence on BAER, but under the conditions of the present study, the effect was not significant.     

应用毛发PCR法检测猕猴TRIM5基因序列CypA转座子插入的TRIMCyp嵌合基因型

为检测猕猴TRIM5基因序列CypA转座子插入TRIMCyp突变基因型个体,本研究建立了应用毛发PCR方法,在基因组水平初步检测猕猴属中TRIMCyp基因型方法,并从猕猴属食蟹猴中初步筛选出TRIMCyp突变基因杂合和纯合子个体。实验以平顶猴为阳性对照,无CypA插入的PCR扩增产物片段大小约为2 343 bp,纯合体为3 110 bp,而杂合突变体则为2 343 bp和3 110 bp。与从血液中提取基因组方法检测结果一致。该方法的建立,为进一步调查TRIMCyp嵌合基因型猕猴在中国的分布,以及研究逆转录病毒跨种间传播和建立HIV-1易感猕猴属动物模型提供了实验手段和平台。     

Primary and secondary tumours occurring simultaneously in the brain of a dog

Primary brain tumours of a single histological type and metastatic brain tumours are well described in dogs in the current veterinary literature. However, the concurrent presence of a primary and secondary tumour in the brain of a dog has never, to the authors' knowledge, been previously reported. The clinical and pathological features of a nine-year-old, female boxer with an oligodendroglioma and metastases from a mammary gland adenocarcinoma occurring simultaneously in the brain are described in this case report. Information in the veterinary literature on multiple malignancies affecting the central nervous system is very limited; therefore, a discussion about comparative situations in human medicine has been included.     

Apoptosis and mitosis in tumours of the skin and subcutaneous tissues of the dog

The presence of apoptotic cell death was evaluated in routinely processed tissue samples of 39 neoplasms of the skin and subcutaneous tissues of the dog using the method of terminal deoxynucleotidyl transferase (T d T) mediated deoxyuridine-5'-triphosphate (d UTP)-biotin nick end labelling (TUNEL). The degree of apoptosis was related to the frequency of mitosis, an index of cell proliferation. The correlation between the apoptotic index (AI), the percentage of positive cells after randomly enumerating 1000 cells and the mitotic count (MC), the number of mitotic figures in 10 fields at a magnification of 400 times was assessed by the Spearman non-parametric correlation test. TUNEL signals were observed in all types of tumours as brown products detected in non-pyknotic nuclei, in non-identifiable rounded structures (so-called apoptotic bodies) and occasionally in the cytoplasm, either singly or in combination. An inverse relationship between AI and MC was observed in benign tumours, while no correlation was found between AI and MC in either malignant or locally invasive tumours. Among benign tumours, intracutaneous cornifying epithelioma, fibroma, haemangioma and Schwannoma had high AI and low MC, while histiocytomas had low AI and high MC and pilomatrixomas low AI and MC. All malignant tumours had low AI and high MC, except for fibrosarcomas, which had high AI and MC. Finally, higher heterogeneity was observed among locally invasive tumours, as they had high AI and low MC (squamous cell carcinomas), and low AI with either low MC (haemangiopericytomas) or high MC (basal cell tumours). The classification of the tumours according to their AI (>15.8% high and <15.8% low) and MC (>9 high, <9 low) did not reflect the clinical behaviour of some tumour types.