Identification and Transfer of Free Cutaneous Flaps by Microvascular Anastomosis in the Dog

An axial pattern cutaneous free flap in the dog based on superficial cervical vessels was identified. Consistent anatomic landmarks were proposed. The vessels consistently emerged from the angle formed by the omotransversarius and trapezius muscles, and coursed in a craniodorsal direction. Eight orthotopic transfers were performed to evaluate the flap's potential as a free tissue transfer. Six of eight (75%) flaps survived. Arterial insufficiency was the likely cause of one failure. The cause of the other failure was considered to be venous occlusion. We conclude that the superficial cervical flap can be used successfully as a microvascular free tissue transfer in the dog.     

Transfer of Free Vascular Cutaneous Flaps by Microvascular Anastomosis Results in Six Dogs

Skin defects on the distal extremities of six dogs were reconstructed with free vascular cutaneous transfers by microvascular anastomosis. The donor flaps were based on the superficial cervical artery and vein. In five of the dogs, bone was exposed and skin was lost from half of the circumference of the limb. Two had infected fractures with sequestra and three had acute shearing injuries. The sixth dog had sensory denervation of the left antebrachium and a carpal acral lick granuloma. Before surgery, the patency of potential recipient vessels was confirmed with arteriography in five dogs and an ultrasonic doppler in one dog. Microvascular technique was used to reestablish circulation to the flaps after they were transferred to the recipient site. Total ischemic time of the flaps averaged 100 minutes. All flaps survived. Successful reconstruction of the cutaneous defects was achieved in these six cases.     

猪繁殖与呼吸综合征病毒-脂多糖刺激对中性粒细胞黏附猪肺微血管内皮细胞的影响

【目的】 研究猪繁殖与呼吸综合征病毒(PRRSV)单独和联合脂多糖(LPS)刺激对肺脏募集中性粒细胞(Neu)的影响, 以及猪肺微血管内皮细胞(MVECs)在其中的作用。【方法】 取约10日龄仔猪的肺脏组织, 采用Ⅱ型胶原酶消化和差速贴壁法, 分离培养原代MVECs, 采用密度梯度离心法分离猪外周血Neu; 以PRRSV HN株或PRRSV-LPS刺激Neu和MVECs后, 将Neu加入培养MVECs的培养板, 孵育1 h, 4%多聚甲醛固定, 瑞氏染色, 观察并计数分析各组黏附的Neu数量; 采用免疫细胞化学染色法检测MVECs对P-选择素和E-选择素的表达; 采用虎红溶液染色法测定分析P-选择素和E-选择素抗体封闭MVECs时, PRRSV HN株或PRRSV-LPS刺激对Neu黏附于MVECs数量的影响。【结果】 分离培养的MVECs呈CD34免疫荧光染色阳性, 阳性率约为92%;PRRSV HN株刺激18 h, 黏附于MVECs的Neu数量显著增加(P<0.05);PRRSV-LPS联合刺激时, 黏附的Neu数量显著多于LPS单独刺激(P<0.05);PRRSV或PRRSV-LPS刺激Neu和MVECs二者时, 黏附Neu数量的增加比单独刺激MVECs或Neu更明显; 免疫细胞化学染色显示, MVECs强阳性表达P-选择素和E-选择素; 用P-选择素和E-选择素抗体封闭MVECs时, PRRSV或PRRSV-LPS刺激诱导的Neu黏附增加呈不同程度下降, E-选择素封闭时具有显著差异(P<0.05)。【结论】 PRRSV感染能促进Neu与MVECs黏附, 亦能增加后继LPS刺激时黏附的Neu数量, MVECs表达的E-选择素是介导PRRSV致Neu黏附增加的重要分子之一。     

Myoperitoneal Microvascular Free Flaps in Dogs: An Anatomical Study and a Clinical Case Report

The purpose of the anatomical study was to identify potential myoperitoneal microvascular free flaps, in dogs, that are based on a single artery and vein. The angiosomes of the right deep circumflex iliac artery and left phrenicoabdominal (cranial abdominal) artery were evaluated in six medium-sized canine cadavers. The right deep circumflex iliac artery and left phrenicoabdominal (cranial abdominal) artery were injected with a mixture of barium and latex (equal parts). The entire right and left transversus abdominis muscles were dissected from the abdominal wall and radiographed. The angiograms of the deep circumflex iliac artery showed poor arborization of the vessels within the transversus abdominis muscle in all six cadavers. The angiograms of the phrenicoabdominal (cranial abdominal) artery showed consistent filling of the vascular bed of the cranial half of the transversus abdominis muscle flap in all six dogs. The vascular pedicle lengths and the diameter of the arteries and veins of both the deep circumflex iliac and phrenicoabdominal (cranial abdominal) myoperitoneal free flaps were found to be acceptable for microvascular anastomosis. The deep circumflex iliac flap was unacceptable because of inadequate vascular perfusion. The cranial abdominal artery had a consistent, large branch that supplied the cranial half of the transversus abdominis muscle, thereby making a myoperitoneal flap supplied by this vessel a potentially useful free flap. An 8-year-old male, neutered, mixed-breed dog was evaluated for possible repair of a large defect of the hard palate. Previous operations, using local tissue flaps, had been unsuccessful. A myoperitoneal free flap, based on the right cranial abdominal artery, and consisting of the cranial portion of the transversus abdominis muscle, was used successfully to reconstruct the hard palate. Migrating epithelium from the edges of the wound covered the myoperitoneal flap by 10 weeks after surgery. Therefore, the cranial abdominal myoperitoneal free flap can be considered for reconstruction of intra-oral defects that cannot be repaired using conventional local flap techniques.     

Transfer of Deep Circumflex Iliac Flaps to the Tarsus by Microvascular Anastomosis in the Horse

Eighteen deep circumflex iliac flaps were elevated in healthy adult horses. Four flaps survived well when elevated as islands, but five orthotopic flaps and nine heterotopic flaps transferred to the tarsus and face failed. Technical reasons could explain the failure of the orthotopic flaps, but not the heterotopic flaps. Failure of the heterotopic flaps was apparently caused by the no-reflow phenomenon.     

ZD制剂对马肢蹄软组织损伤部位肌组织形态及VEGF和bFGF表达的影响

ZD制剂为治疗马非开放性软组织损伤的经典验方,为探索其作用机制,通过建立马前肢急性软组织损伤模型,对其进行组织形态学观察,评价ZD制剂对马急性软组织损伤的作用效果,采用免疫组织化学方法检测ZD制剂对VEGF、bFGF在肌肉中表达的影响。用20匹蒙古马随机分为4组,选取左前肢肩部臂部肌肉,制造急性软组织损伤模型。于造模前、后用取样器进行组织取样,制作成石蜡切片,观察其形态学变化,用免疫组织化学方法对阳性细胞分析VEGF、bFGF的表达。结果表明,试验药物组在损伤后的修复能力优于阳性药物组和阴性对照组。表明ZD制剂可通过促进成纤维细胞、毛细血管新生和肌细胞的生成来进行组织的修复再生,试验药物组在损伤后VEGF、bFGF的表达优于阴性对照组和阳性药物组,ZD制剂可能通过提高VEGF、bFGF的表达进行损伤的修复。     

Triiodothyronine Has No Enhancement Effect on the Osteogenic or Chondrogenic Differentiation of Equine Adipose Tissue Stem Cells

The effects of two concentrations of triiodothyronine (T3; 0.01 and 1,000 nM) on the osteogenic and chondrogenic differentiation abilities of equine adipose-derived mesenchymal stem cells (AD-MSCs) were evaluated. The osteogenic study evaluated the effect of T3 using alkaline phosphatase activity (ALP) assay; cell viability and density; and formation of mineralized nodules at Days 7, 14, and 21 in culture. The chondrogenic study tested the effect of T3 through ALP assay, mitochondrial metabolism, cell density, and periodic acid–Schiff-positive (PAS+) matrix percentage at Days 7 and 14. In both experiments, analysis of variance was used to compare averages through the Student–Newman–Keuls test. In the osteogenic study, no differences in any variable were detected between groups at Day 7. At Day 14, 0.01 nM T3 reduced cell density and the number of mineralized nodules despite the increase in ALP activity and mitochondrial metabolism (P < .05). ALP activity increased at 1,000 nM T3 concentration (P < .05). At Day 21, 0.01 nM T3 treatment increased ALP activity compared with control treatment (P < .05). At 1,000 nM concentration, T3 reduced mitochondrial metabolism and cell density (P < .05). In the chondrogenic study, the two T3 concentrations increased cell density compared with control treatment at Day 7. At Day 14, higher T3 concentration reduced mitochondrial metabolism, ALP activity, cell density, and PAS+ chondrogenic matrix percentage compared with control treatment (P < .05). Thus, T3 addition to equine AD-MSC cultures has no enhancement effect on osteogenic or chondrogenic differentiation and may, in fact, negatively affect cell density and matrix synthesis depending on hormone concentration and culture time.     

Preliminary Notes on Equine Tissue Transglutaminase Serology and A Case of Equine Gluten-Sensitive Enteropathy and Dermatitis in an 11-Year-Old Dutch Warmblood Horse

It has been suggested that gluten may play a role in equine inflammatory small bowel disease (ISBD). Previous work showed an association between equine gluten-sensitive enteropathy and IgA antibodies to tissue transglutaminase (TGA) in serum. The purpose of this study is to investigate the prevalence of IgA antibodies to TGA in a group of healthy non–gluten-free sport ponies and to present a case of tentative gluten-sensitive enteropathy and dermatitis in a horse. Blood samples were obtained from 40 healthy jumping ponies. The ponies comprised 12 mares, 8 stallions, and 20 geldings with an average age of 9.0 ± 3.8 years (±SD; range 3–19 years). Sera were tested for IgA antibodies against human recombinant TGA. Significance (P < .05) of the correlation between TGA titer and age in these ponies was assessed using Pearson test (two tailed). In addition, to further illustrate tentative equine gluten-sensitive enteropathy and dermatitis, the clinical course in an 11-year-old Dutch Warmblood sport horse gelding has been described. The average TGA titer was 21.4 ± 13.6 AU/mL (range 2–65 AU/mL). There was a significant (P = .013) correlation (r = 0.389) between age and TGA titer in ponies. One of the 40 ponies (2.5%) showed an elevated TGA titer. An elevated TGA titer decreased after a gluten-free ration for 3 months in an 11-year-old Warmblood gelding with a tentative diagnosis of ISBD associated with full remission of the generalized skin reaction. To our best knowledge, this is the first study assessing TGA antibodies in sera from healthy non–gluten-free ponies and showing a correlation with age. The presented case could be the first one of a horse with a tentative diagnosis of gluten-sensitive enteropathy combined with dermatitis. Given the reported findings, this study warrants further investigations into gluten-sensitive enteropathy and dermatitis in individual horses affected with ISBD.     

Microvascular Free Tissue Transfer: Results in 57 Consecutive Cases

Objective —To evaluate the outcomes and complications in a consecutive series of animals undergoing microvascular reconstructive procedures at two veterinary institutions. Study Design—Retrospective study. Animals or Sample Population—A total of 44 client-owned dogs and one red-necked wallaby. Methods —The medical records of all animals undergoing reconstructive microsurgical procedures at the Western College of Veterinary Medicine and Michigan State University were reviewed. Microvascular flap survival and related complications were described. Statistical analysis was performed to determine the significance of relationships between operative factors and outcome. Results —A total of 57 microvascular procedures were performed on 55 animals. Reconstruction was required after trauma in 42 animals, after ablative cancer surgery in 11 animals and for correction of congenital tissue aplasia in 1 animal. Donor tissues included the superficial cervical cutaneous, medial saphenous fasciocutaneous or musculofasciocutaneous, caudal superficial epigastric cutaneous, trapezius muscle or musculocutaneous, caudal sartorius muscle, latissimus dorsi muscle or musculocutaneous, cranial abdominal myoperitoneal, carpal footpad, digital footpad, and vascularized ulnar bone flaps. A total of 53 of 57 flaps (93%) survived. There was a significant relationship between flap failure and level of assistant surgeon experience (P < .05). Latissimus dorsi flaps were significantly more likely to fail when compared with pooled data from all other flap types (P < .01). Conclusions —The success of microvascular tissue transfer in this case series compares favorably with those reported in human reconstructive microsurgery. Both the primary and assistant surgeon should be practiced in microsurgical technique. Failure of latissimus dorsi flaps was not likely caused by an inherently deficient flap design, but was more likely attributed to the location and severity of trauma at the recipient site, the difficulty in isolating suitable recipient vessels for anastomosis or the absence of a trained assistant surgeon during these procedures. Clinical Relevance —This retrospective study documents the successful application of microvascular technique in a series of clinical cases requiring tissue reconstruction.     

Experiences and Skills for Establishment of Rat Model of Myocardial Ischemia and Reperfusion

The establishment of rat model of myocardial ischemia and reperfusion is a very sophisticated experimental process. During animal experiments, beginners always meet with troubles inevitably. This paper have conducted a great deal of this type of experiments for years, and intend to share some experiences and lessons with colleagues.     

Equine Endometrial Tissue Concentration of Fluconazole Following Oral Administration

The objective of this study was to determine the plasma and endometrial tissue concentrations of orally administered fluconazole and to determine if these tissue levels surpassed the minimum inhibitory concentration (MIC) for Candida spp. organisms in the reproductive tract of the mare. Mares from study 1 (n = 9) were administered a single oral loading dose of 14 mg/kg fluconazole. Plasma and endometrial tissue samples were collected before fluconazole administration and for 24 hours after the loading dose. Study 2 mares (n = 3), a subset of study 1, were administered the loading dose, followed by maintenance doses of 5 mg/kg every 24 hours for 6 days. Plasma and biopsy samples were collected for 48 hours after the last maintenance dose. High pressure liquid chromatography-tandem mass spectrometry was used to determine the concentration of fluconazole in all samples. The mean plasma and endometrial fluconazole levels 24 hours after the loading dose were 9.53 ± 0.824 μg/mL (mean ± standard deviation) and 11.3 ± 2.38 μg/g, respectively. Fluconazole levels in plasma and endometrial tissue 24 hours after the last maintenance dose were 7.82 ± 1.81 μg/mL and 7.23 ± 3.86 μg/g, respectively. Oral fluconazole administered as a 14-mg/kg loading dose and a 5-mg/kg maintenance dose every 24 hours will result in endometrial tissue levels near the accepted MIC values for most Candida spp. and surpass the MIC for Candida albicans in the reproductive tract of the mare. Consequently, this dosage regimen could be considered for the treatment of infectious endometritis caused by susceptible fungal organisms of Candida spp. in the mare.     

6-苄氨基嘌呤对大鼠小肠缺血再灌注损伤的影响

本试验旨在研究6-苄氨基嘌呤(6-BA)对大鼠小肠缺血再灌注(I/R)损伤的保护作用。选取80只雄性SD大鼠,随机分为4组,即对照组、I/R模型组和低、高剂量6-BA组。低、高剂量6-BA组于术前3周分别连续灌胃10、20mg/kg的6-BA,对照组和I/R模型组灌胃同体积的生理盐水,每天1次。对照组在暴露肠系膜上动脉后不做阻断;I/R模型组和低、高剂量6-BA组阻断肠系膜血管30min后再灌注60min。随后取大鼠空肠组织分别进行总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量检测;采用单细胞凝胶电泳法检测细胞DNA损伤程度,采用免疫组化法检测半胱氨酸天冬氨酸特异性蛋白酶3(Caspase-3)表达情况。结果显示,与I/R模型组相比,补充10、20 mg/kg的6-BA后,T-SOD、GSH-Px活性显著升高(P0.05),MDA含量显著降低(P0.05);小肠细胞DNA的拖尾现象好转,尾部DNA含量和拖尾率都显著低于I/R模型组(P0.05);Caspase-3阳性表达细胞数量显著减少(P0.05)。由此可见,10、20 mg/kg的6-BA能有效防护I/R对小肠的损伤,且20mg/kg的6-BA作用效果尤为明显。     

Microvascular Free Tissue Transfer of the Rectus Abdominis Muscle in Dogs

OBJECTIVE: To assess donor-site morbidity and survival of the rectus abdominis muscle with an overlying skin graft after free tissue transfer to a medial femorotibial defect in dogs. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Phase 1: 6 canine cadavers; phase 2: 7 adult mixed-breed dogs. METHODS: Phase 1: The rectus abdominis muscle was removed from canine cadavers, muscular and vascular dimensions were recorded, and angiography was performed. Phase 2: Muscular transfer was performed through anastomosis of the caudal epigastric artery and vein to the saphenous artery and medial saphenous vein. Transferred tissues were evaluated on postoperative days 3, 6, 10, and 13. Animals were examined daily until euthanasia between postoperative days 31 and 42. Postmortem angiograms were performed and tissues collected for histopathologic evaluation. RESULTS: Phase 1: Appropriate vascular dimensions for microvascular anastomosis were confirmed and surgical technique perfected. Phase 2: Muscular excision produced minimal donor-site morbidity. All muscles survived after microvascular transfer and angiography confirmed vascular patency. All of the skin grafts survived, with one graft undergoing partial necrosis. CONCLUSIONS: The rectus abdominis muscle can be successfully transferred to a medial femorotibial defect and can serve as a bed for acute skin grafting. No significant donor-site morbidity is associated with its removal. CLINICAL RELEVANCE: Microvascular free tissue transfer of the canine rectus abdominis muscle has not been previously described. This technique provides a new alternative for repair of appropriate wounds. Additional studies are needed to define its utility in clinical patients.     

家兔脑缺血再灌注损伤后边缘系统一氧化氮合酶活性的变化及血塞通对其变化的影响

为了研究家兔脑缺血再灌注损伤后边缘系统一氧化氮合酶(NOS)活性的变化及血塞通对其活性变化的影响,探讨血塞通对全脑缺血家兔的脑保护作用.选取3月龄哈白兔63只,体质量(1500±150)g,随机分为脑缺血治疗组,脑缺血未治疗组和对照组3组,手术建立家兔全脑缺血模型,应用生化检测技术测定脑缺血再灌注后不同时间边缘系统NOS活性变化及血塞通对其活性的影响.结果表明,边缘系统内NOS活性在脑缺血再灌注2 h后开始升高,6 h达到高峰,随后逐渐下降,24～96 h活性持续下降,120 h后恢复至正常水平;脑缺血治疗组下降幅度大、速度快,在96 h即恢复至正常水平.缺血治疗组和正常对照组NOS的活性极最著低于缺血未治疗组(P<0.01).结果提示,血塞通可以通过减弱NOS活性进而维持NO的生理含量,对全脑缺血再灌注损伤家兔有明显的脑保护作用.     

Tissue Culture of the Enteric Nervous System from Equine Ileum

Ileal samples were harvested fresh from euthanized adult horses. The tissues were microdissected to prepare wholemount preparations for immunohistochemistry and for either explant or dissociated culture systems of the enteric nervous system. Explant culture systems were established using wholemounts of either the submucous plexus or the muscularis externa (including the myenteric plexus). Dissociated cell cultures could only be obtained from the submucous plexus. Culture systems were maintained for up to 5 days. Immunoreactivity for a neuronal marker (Pan-N) and for glial cell markers (GFAP and S100) indicated the presence of both neurons and enteric glia in the tissue culture preparations.This is the first report of equine enteric neurons being grown in tissue culture. Further refinements to the techniques will be required before this in vitro model can be used for quantitative analysis.     

The Effect of Tension on Perfusion of Axial and Random Pattern Flaps in Foals

Caudally based axial pattern and random pattern skin flaps, 5 cm x 10 cm, were raised on the abdomens of seven foals. Blood flow was measured 7.5 cm from the flap bases during incremental increases in applied tension. At tensions less than 1400 g, cutaneous blood flow was more than four times higher in axial pattern flaps than in random pattern flaps. Blood flow in axial pattern flaps at tensions up to 1500 g was greater than baseline flow in random pattern flaps. After removal of applied tension, perfusion in axial pattern flaps gradually increased but remained below baseline values. A short period of hyperperfusion after removal of tension in random pattern flaps may have been indicative of accumulation of waste products and localized acidosis. All flaps healed in their donor beds without complications.