鹿复合麻醉剂对鹿生理指征、ANP和CGRP的影响

研究鹿复合麻醉剂对鹿生理指征、血浆心钠素(ANP)及降钙素基因相关肽(CGRP)的影响,以明确药物对机体作用,进而指导药物的临床应用。试验选取6只健康成年梅花鹿,肌注鹿复合麻醉剂0.04 mL/kg,注药前及注药后15、30、45、60、75、90及120 min进行收缩压(SBP)、舒张压(DBP)、平均动脉压(MAP)和心率(HR)的监测,并同步采集颈静脉血样,酶联免疫吸附法测定血浆中ANP和CGRP含量。结果表明,鹿复合麻醉剂引起麻醉期鹿血浆ANP和CGRP含量变化,使血管发生舒缩改变,导致SBP、DBP、MAP和HR产生相应的变化,苏醒后各指标均恢复到麻前水平。该药物可以应用于鹿的临床麻醉。     

Gross anatomical study of the thoracolumbar primary dorsal branches and their course from the spinal cord to the skin in the cat

The course of spinal nerves and the corresponding cutaneous areas are fundamental for numerous therapeutic approaches used in complementary veterinary medicine. Positive effects of these methods are primarily based on segmental reflex arcs which are associated with the course of the spinal nerves. In this morphological study, the lateral cutaneous branches of the thoracolumbar dorsal branches from Th9 to L7 were examined in cats with special regard to their anatomical course. A four-layer dissection was carried out to reveal the course of nerves between the intervertebral foramina and their point of entry into the skin, starting in the dorsal midline. Dorsal branch courses and covered distances were documented and measured in each layer. The covered distance was evaluated by the Caudal Shift Index (CSI_n) on both body sides and within each layer. The ‘back region’ was used as relative dimensional unit, describing the distance between the cranial tips of two consecutive spinous processes. Overall, the mean CSI_n for dorsal branches of Th9 to L7 amounted to three back regions from the intervertebral foramen to the skin entry point of a dorsal nerve branch. This provides therapists with clues and should be put into practice, by extending the treatment area up to three segments caudally from the nerve exit point. Furthermore, the results of this study present new data on inferred lumbar dermatomes in cats, data which until now have only been transferred from other species. These results may serve as an anatomical foundation for manual therapies.     

Immunohistochemical localization of calcium binding proteins and some neurotransmitters in myenteric plexus of goat stomach

To understand the neurochemical properties of the gastric myenteric plexus of ruminants, the expression patterns of calbindin D-28k (CB), calretinin (CR), substance P (SP) and calcitonin gene-related peptide (CGRP) were explored in the Korean native goat. In gastric myenteric plexus, CB and SP immunoreactivity were observed in round- or oval-shaped neurons. CR and CGRP immunoreactivity were detected only in the nerve fibers. This immunohistochemical localization of CB, CR, CGRP and SP in the myenteric plexus of the goat stomach exhibited species-specific patterns. These findings suggest that these substances may be directly or indirectly related to the gastric functions of the goat stomach.     

ORIGINAL ARTICLE: The approach to the mechanism of calcitonin gene‐related peptide‐inducing inhibition of food intake

The aim of this study was to investigate the anorectic mechanism of calcitonin gene‐related peptide (CGRP) in rats. Intraperitoneal injection of CGRP (50 μg/kg) resulted in decline (p < 0.05) in the food intake of rats at 0.5, 1, 2 and 4 h in comparison with saline control. Compared with saline‐treated group, the levels of hypothalamic 3′,5′‐cyclic adenosine monophosphate (cAMP) and plasma glucagon were increased (p < 0.05) in CGRP‐treated group, but insulin level was decreased (p < 0.05). No significant changes (p > 0.05) in the plasma leptin were observed between two treatment groups. Calcitonin gene‐related peptide injection down regulated (p < 0.05) both neuropeptide Y (NPY) and melanin‐concentrating hormone (MCH) genes at mRNA levels, but up regulated (p < 0.05) the expression of cholecystokinin (CCK) gene. The correlations analysis showed that food intake was negatively correlated (p < 0.05) with CCK mRNA, cAMP and glucagon levels. Moreover, there existed negative correlations (p < 0.05) between MCH mRNA and glucagon levels, and positive correlations (p < 0.05) between insulin and leptin levels. The results showed that cAMP acting as the second messenger may play a vital role in the anorectic effects of CGRP. Calcitonin gene‐related peptide could stimulate anorexigenic neuropeptides (i.e. CCK) and/or inhibit orexigenic neuropeptides (i.e. NPY and MCH) expression, and ultimately suppressed food intake that was functionally coupled to cAMP/PKA pathway activation.     

Effect of active immunization against GnRH‐I on the reproductive function in cat

This study was designed to explore the effect of active immunization against maltose binding protein‐gonadotropin releasing hormone I hexamer (MBP‐GnRH‐I6) on the reproductive function in cats. Each immunized cat was administered twice intramuscularly in the neck at 16 and 20 weeks old. The concentrations of the testosterone and estradiol and the level of anti‐GnRH‐I antibody in the serum were measured by radioimmunoassay and ELISA, respectively. The results showed that the weight and size of testicles and ovaries, and the concentrations of serum testosterone and estradiol in the immunized animals were lower than those of the control cats (P < 0.05), but that the levels of anti‐GnRH‐I antibody were significant higher compared to control animals (P < 0.05). Testicular tissues from the immunized male cats showed that seminiferous tubules were depauperate with the lumen relatively empty and that the differentiation of spermatogonia was not obvious. Tissues from the immunized female cats showed that the ovaries had many primordial follicles and primary follicles, but no secondary follicle was observed. These results showed active immunization against MBP–GnRH‐I6 could make the gonads atrophy and reduce the concentrations of gonadal hormones, which suggested that MBP‐GnRH‐I6 was a very effective immunogen in the cat.     

Duration of corneal anaesthesia following multiple doses and two concentrations of tetracaine hydrochloride eyedrops on the normal equine cornea

Reasons for performing study: There is a clinical impression that tetracaine hydrochloride (THCl) eyedrops is a suitable topical anaesthetic in horses. Objective: To determine the duration of corneal anaesthesia following instillation of multiple doses and 2 concentrations of THCl in 10 healthy horses. Methods: The corneal touch threshold (CTT) was determined, in both eyes, before (basal CTT) and after application of one drop of 0.5%THCl, 2 drops at a 1 min interval of 0.5%THCl or one drop of 1%THCl. CTT was measured in mm every 5 min until complete recovery of the basal CTT. Treatments were separated by an interval of at least one week. Results: Corneal sensitivity was significantly reduced from baseline values for 30, 60 and 50 min after application of one drop of 0.5%THCl, 2 drops of 0.5%THCl and one drop of 1%THCl, respectively. Mean maximal anaesthetic effects, corresponding to a CTT of 0 mm, lasted 5.5, 16 and 15.25 min and maximal anaesthetic effect was present in 55, 90 and 80% of eyes, 5 min after application of one drop of 0.5%THCl, 2 drops of 0.5%THCl and one drop of 1%THCl, respectively. Conclusions: The application of a second drop or the use of more concentrated eyedrops significantly increases duration of both anaesthesia and maximal anaesthetic effect. Potential relevance: Duration of corneal anaesthesia following tetracaine instillation was established enabling a better use when performing diagnostic and therapeutic procedures. Comparison of tetracaine with other ocular anaesthetics needs to be published in the future.     

Differences between µCT‐imaging and conventional CT for the diagnosis of possible diseases of the middle and inner cat ear

The aim of this study was to check the relevance of using in‐vivo micro computed tomography (µCT) for the diagnosis of possible diseases of the middle and inner ear of the cat. Therefore, on the one hand, differences of the detail detectability between the two imaging methods conventional computed tomography (cCT) and in‐vivo µCT were analyzed. Six healthy cat ears were dissected and scanned several times and the obtained images were compared with each other. On the other hand, histological slices of all ears were prepared and pictures of defined anatomical structures were taken and compared with the identical sectional plane of the µCT‐images. This way it was possible to evaluate the quality and clinical limitations of the in‐vivo µCT. The results show that an in‐vivo µCT is suitable to analyze even the smallest osseous structures, such as the semicircular ducts, the spiral osseous lamina or the ossicles whereas with the help of cCT it is not possible to identify such small osseous structures because of their blurred and less detailed representation. Delicate soft tissue structures as the membranous labyrinth including hearing and vestibular organ cannot be differentiated with as well in‐vivo µCT‐ as with cCT‐images. In‐vivo µCT represent a good possibility for more detailed diagnosis of extremely fine structures which cannot be detected with cCT. Histological slices can nonetheless not be replaced by in‐vivo µCT due to a too low spatial resolution and the limitations of the in‐vivo µCT with regard to the evaluation of soft tissue dense structures.     

Modulation of electrogenic transport processes in the porcine proximal colon by enteric neurotransmitters

The aim of our study was to evaluate the involvement of essential pro- and antisecretory neurotransmitters in regulation of secretion in porcine proximal colon. Choline acetyltransferase (ChAT), nitric oxide synthase (NOS), vasoactive intestinal peptide (VIP), substance P (SP), somatostatin (SOM) and neuropeptide Y (NPY) were located immunohistochemically in the epithelium and subepithelial layer. Modulation of epithelial secretion was studied in Ussing chambers. Application of carbachol (CA), sodium nitroprussid (SNP), VIP and SP but not of NPY or SOM resulted in a chloride dependent increase in short circuit current (I(sc) ). I(sc) increase induced by CA, VIP or SNP was not altered by preincubation with tetrodotoxin or indomethacin. In contrast, SP-induced I(sc) increase was diminished by preincubation with tetrodotoxin, indomethacin, L-nitro-arginin-methyl-ester, and atropine but not hexamethonium. Simultaneous application of CA and VIP, or CA and SNP increased the I(sc) stronger as expected. Applying SP/CA led to a smaller increase in I(sc) as calculated. It is concluded that mainly prosecretory neurotransmitters are involved in regulation of colonic secretion. Cross-potentiations of acetylcholine and nitric oxide and acetylcholine and VIP suggest activation of different intracellular cascades. Similar intracellular pathways may be stimulated by acetylcholine and SP, thus preventing an additive effect of the transmitters.     

Immunohistochemical analysis of pseudorabies virus spread through neurons innervating the eyeball

Pseudorabies virus (PRV) was inoculated intraocularly into BALB/c mice, and the distribution pattern of cells positive for several neurotransmitters and viral antigens in the eyeball, trigeminal nerve ganglia, and superior cervical ganglia was examined immunohistochemically to clarify the neural route of the virus spread. In the eyeball, substance P (SP)- and calcitonin gene-related peptide (CGRP)-positive cells were detected in the ipsilateral iris and ciliary body, neuropeptide tyrosine (NPY)-positive cells were detected in the choloid membrane, and tyrosine hydroxylase (TH)-positive cells were detected in the ipsilateral inner nuclear layer of the retina; all these cells contained viral antigens. In the superior cervical ganglia, viral antigen-positive cells containing TH or NPY were found at bilateral sites. In the trigeminal nerve ganglia, viral antigen-positive cells containing SP or CGRP were found at bilateral sites. These findings indicated that the SP- and CGRP-positive ganglion cells of the trigeminal nerve ganglia innervating the iris or ciliary body, and the NPY-positive ganglion cells of the superior cervical ganglia innervating the iris, ciliary body, and choroid membrane served as the route for the virus spread. These findings also suggested that dopaminergic neurons, such as the TH-positive retinal cells and TH-positive ganglion cells of the superior cervical ganglia, served as the route for virus spread.     

Medication‐related osteonecrosis of the jaw after long‐term bisphosphonate treatment in a cat

A 12‐year‐old, neutered female, domestic medium hair cat was evaluated for a nonhealing, oral mucosal ulceration. The cat had a history of idiopathic hypercalcemia that had been treated with a bisphosphonate for 41 months. Oral examination identified exposed maxillary bone adjacent to a previous extraction site. Histopathology of the exposed bone and associated mucosa was most consistent with medication‐related osteonecrosis of the jaw. Treatment involved both medical and surgical interventions. Oral mucosal healing occurred after 6 months of treatment.     

潮霉素抗性基因筛选标记质粒的构建及LL37的表达

为了实现以潮霉素磷酸转移酶基因(hpt)为选择标记的质粒在毕赤酵母中高效表达抗菌肽LL37,在pGAPZαA质粒的基础上,通过EcoRⅤ、BamHⅠ双酶切将Zeocin抗性基因替换成潮霉素磷酸转移酶基因,获得的表达质粒命名为pGAPHαA。根据LL37氨基酸序列,通过毕赤酵母密码子偏嗜性改造,合成的LL37基因片段克隆到pGAPHαA质粒中,获得重组分泌型表达质粒pGAPHαA-LL37,线性化后电击转化毕赤酵母SMD1168,经潮霉素B筛选阳性转化子。72 h表达上清LL37蛋白含量约为167 μg /mL,表达产物耐热性好,对大肠杆菌DH5α、猪链球菌最小抑菌浓度分别为2.94、7.06 μg/mL。     

Properties of the feline tumour suppressor reduced expression in immortalized cells (REIC/Dkk‐3)

Reduced expression in immortalized cells (REIC/Dkk‐3), a member of the human Dickkopf (Dkk) family, is a growth suppressor in human and canine mammary tumours. Mammary gland tumours are common neoplasms with high malignancy in female cats. The purpose of this study was to clone the feline REIC/Dkk‐3 homolog, investigate its expression in cell lines established from feline mammary gland tumours, and test its tumour suppressor function. Western blot analysis revealed that expression of the REIC/Dkk‐3 protein was reduced in feline mammary carcinoma cell lines. Forced expression of REIC/Dkk‐3 induced apoptosis in feline mammary tumour cell lines. These results demonstrate that REIC/Dkk‐3 expression, which is downregulated in feline mammary tumour cell lines, results in the induction of apoptosis in these cells. Our findings suggest that feline REIC/Dkk‐3 represents a potential molecular target for the development of therapies against feline mammary cancers.     

Distribution of the neurokinin-1 receptor in equine intestinal smooth muscle

REASON FOR PERFORMING STUDY: Tachykinins have profound effects on equine intestinal motility, but the distribution of the neurokinin receptors (NKRs) through which they act is unknown. This study reports the distribution of one of these receptors, the neurokinin-1 receptor (NK1R), in smooth muscle throughout the equine intestinal tract. OBJECTIVES: To quantify the distribution of the NK1R, based upon mRNA expression, in smooth muscle of different regions of the equine intestinal tract. METHODS: Nine regions of the intestinal tract were sampled in 5 mature horses. Total RNA was isolated from smooth muscle and reverse transcribed; NK1R mRNA was then quantified using real-time PCR. RESULTS: NK1R mRNA was found at all levels of the sampled intestinal tract. The smooth muscle of the proximal small intestine and the ventral colon exhibited the highest level of NK1R mRNA expression in the equine intestinal tract. CONCLUSIONS: Tachykinins probably affect intestinal contractility and propulsion in the proximal small intestine and in the ventral colon.