Everted third eyelid cartilage in a cat: a case report and literature review

An 8-year-old neutered female British Blue cat was presented with a presumed diagnosis of a prolapsed nictitans gland and associated ocular irritation and epiphora. However, during surgery, the apparent nictitans gland protrusion was determined to be an everted cartilage of the nictitating membrane. The scrolled portion of the cartilage was removed through an incision through the conjunctiva on the bulbar aspect of the third eyelid, as previously described in the dog. This operation resolved the ocular irritation occurring, and the third eyelid returned to its anatomically correct position.     

Mucosa-associated lymphoid tissue lymphoma of the third eyelid conjunctiva in a dog

A 4-year-old, neutered female Cocker Spaniel was presented to the veterinary clinic for protrusion of the left third eyelid. When the third eyelids from both eyes were everted, lobulated masses were present on the bulbar surface. The left third eyelid had a larger protrusion. There was no apparent associated ocular or systemic involvement. The tumor of left third eyelid was removed and referred for histological examination. Histologically, there were proliferations of lymphoid follicles surrounded by lymphoid cells forming a marginal zone. Those lymphoid cells occasionally infiltrated into conjunctival epithelium. A few apoptotic bodies with karyopyknotic and karyorrhexic nuclei were observed in the germinal center of lymphoid follicles. Mitotic figures were rare. On immunohistochemistry, tumor cells expressed CD79a but not CD3. A diagnosis of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) of the third eyelid was established based on the histological and immunophenotypical features. At the 1-year follow-up, there was no evidence of recurrence of the mass at the area of excision of the left third eyelid and the remaining tumor of the right third eyelid was still a similar size. The dog still showed no significant findings, except those of the tumor, and no evidence of systemic involvement. To the authors' knowledge, this is the first reported case of MALT lymphoma of the third eyelid in a dog.     

结膜相关淋巴组织和哈氏腺免疫协同关系的研究Ⅰ．结膜相关淋巴组织缺失对哈氏腺点眼免疫应答的影响

采用ＥＬＩＳＡ、免疫组织化学方法、手术切除结膜相关淋巴组织（Ｃｏｎｊｕｎｃｔｉｖａ－ａｓｓｏｃｉａｔｅｄ Ｌｙｍｐｈｏｉｄ Ｔｉｓｓｕｅ，ＣＡＬＴ）法对结膜相关淋巴组织和哈氏腺（Ｈａｒｄｅｒｉａｎ Ｇｌａｎｄ，ＨＧ）在点眼免疫应答过程中的相互协同关系进行了研究。研究发现，采用外科手术于１日龄切除结 膜相关淋巴组织后切除组点眼免疫后泪液中ＮＤＶ－特异性ＩｇＡ、ＩｇＭ型抗体滴度及哈氏腺中ＩｇＡ、ＩｇＭ     

Mast cell tumour in the third eyelid of a mare

A 4‐year‐old Missouri Fox Trotter mare was referred to the Veterinary Teaching Hospital, Koret School of Veterinary Medicine with a reddish mass protruding from the third eyelid and adjacent palpebral conjunctiva of the left eye. On ophthalmic examination, the mare showed mild blepharospasm, mild mucopurulent discharge and mild corneal oedema in the left eye. Menace response, pupillary light reflexes and intraocular examination were normal. A rounded, fleshy red ulcerated mass, approximately 3 cm in diameter, expanded the edge of the outer surface of the medial third eyelid and conjunctiva of the ventromedial palpebral region. Physical examination was otherwise unremarkable. Based on history and clinical findings, surgical removal was elected. The tumour was excised and the conjunctiva in the area of tumour treated with cryotherapy. Histological evaluation revealed a mast cell tumour on the third eyelid. Nine months follow‐up revealed no recurrence.     

Malignant melanoma of the third eyelid in a horse

An 8‐year‐old grey Quarter Horse gelding was referred for evaluation of a rapidly growing mass associated with the third eyelid of the left eye. A pigmented mass of approximately 2 cm in diameter was palpated and visualised associated with the conjunctival lining of the nictitans. It was not possible to palpate normal nictitans deep to the base of the mass. A full dermatological examination revealed no other melanomas in common sites. Based on the size and rapid growth of the mass, surgical excision and one application of local chemotherapy was performed under general anaesthesia. Histopathology confirmed the diagnosis of malignant melanoma and the presence of clean surgical margins. There was no recurrence at 5 weeks post surgery. To our knowledge, this is the first report of a primary malignant melanoma of the third eyelid in a horse.     

玉屏风多糖对小鼠肠黏膜免疫应答和免疫损伤的调控作用

为研究玉屏风多糖对肠黏膜免疫应答和免疫损伤的调控作用,本试验以黄芪多糖为对照药物,以正常小鼠和免疫抑制小鼠为动物模型,150只SPF小鼠随机分为空白对照组、环磷酰胺免疫抑制对照组、玉屏风多糖治疗组、玉屏风多糖阳性对照组和黄芪多糖对照组,每组30只。多糖剂量为200mg/Kg/d,连续灌服7d,环磷酰胺剂量为80mg/Kg,隔日腹腔注射。检测SIgA、IL-2、TGF-β1和IL-6以及小肠黏膜相关淋巴组织的变化。结果发现,玉屏风多糖和黄芪多糖均可显著提高小鼠SIgA、IL-2、TGF-β1和IL-6水平(P<0.05或P<0.01),小鼠肠上皮细胞增生,杯状细胞增多,肠绒毛长度、宽度和隐窝深度均显著增加(P<0.05或P<0.01),肠上皮细胞间淋巴细胞和黏膜固有层淋巴细胞数量明显增多(P<0.05或P<0.01),PP结体积增大,淋巴细胞增生。与黄芪多糖比较,玉屏风多糖对肠黏膜免疫应答的正向调节作用更为明显。对免疫抑制小鼠,玉屏风多糖可有效拮抗环磷酰胺诱导的SIgA降低(P<0.05),并可通过调节TGF-β1(P<0.05)和IL-6(P<0.01)水平,调控Th1/Th2 漂移状态,缓解环磷酰胺所致的免疫损伤,对免疫抑制小鼠小肠黏膜相关淋巴组织也有明显的保护和恢复作用。结果表明,玉屏风多糖可从多方面调控小鼠肠黏膜的免疫应答,可有效增强肠黏膜免疫功能,缓解和改善免疫抑制小鼠肠黏膜免疫屏障的损伤。     

The development of equine immunity: Current knowledge on immunology in the young horse

The development of equine immunity from the fetus to adulthood is complex. The foal's immune response and the immune mechanisms that they are equipped with, along with changes over the first months of life until the immune system becomes adult‐like, are only partially understood. While several innate immune responses seem to be fully functional from birth, the onset of adaptive immune response is delayed. For some adaptive immune parameters, such as immunoglobin (Ig)G1, IgG3, IgG5 and IgA antibodies, the immune response starts before or at birth and matures within 3 months of life. Other antibody responses, such as IgG4, IgG7 and IgE production, slowly develop within the first year of life until they reach adult levels. Similar differences have been observed for adaptive T cell responses. Interferon‐gamma (IFN‐γ) production by T helper 1 (Th1)‐cells and cytotoxic T cells starts shortly after birth with low level production that gradually increases during the first year of life. In contrast, interleukin‐4 (IL‐4) produced by Th2‐cells is almost undetectable in the first 3 months of life. These findings offer some explanation for the increased susceptibility of foals to certain pathogens such as Rhodococcus equi. The delay in Th‐cell development and in particular Th2 immunity during the first months of life also provides an explanation for the reduced responsiveness of young horses to most traditional vaccines. In summary, all immune components of adult horses seem to exist in foals but the orchestrating and regulation of the immune response in immature horses is strikingly different. Young foals are fully competent and can perform certain immune responses but many mechanisms have yet to mature. Additional work is needed to improve our understanding of immunity and immune regulation in young horses, to identify the preferred immune pathways that they are using and ultimately provide new preventive strategies to protect against infectious disease.     

Attenuation of ischaemic injury in the equine jejunum by administration of systemic lidocaine

REASONS FOR PERFORMING STUDY: Absorption of endotoxin across ischaemic-injured mucosa is a major cause of mortality after colic surgery. Recent studies have shown that flunixin meglumine retards mucosal repair. Systemic lidocaine has been used to treat post operative ileus, but it also has novel anti-inflammatory effects that could improve mucosal recovery after ischaemic injury. HYPOTHESIS: Systemic lidocaine ameliorates the deleterious negative effects of flunixin meglumine on recovery of mucosal barrier function. METHODS: Horses were treated i.v. immediately before anaesthesia with either 0.9% saline 1 ml/50 kg bwt, flunixin meglumine 1 mg/kg bwt every 12 h or lidocaine 1.3 mg/kg bwt loading dose followed by 0.05 mg/kg bwt/min constant rate infusion, or both flunixin meglumine and lidocaine, with 6 horses allocated randomly to each group. Two sections of jejunum were subjected to 2 h of ischaemia by temporary occlusion of the local blood supply, via a midline celiotomy. Horses were monitored with a behavioural pain score and were subjected to euthanasia 18 h after reversal of ischaemia. Ischaemic-injured and control jejunum was mounted in Ussing chambers for measurement of transepithelial electrical resistance (TER) and permeability to lipopolysaccharide (LPS). RESULTS: In ischaemic-injured jejunum TER was significantly higher in horses treated with saline, lidocaine or lidocaine and flunixin meglumine combined, compared to horses treated with flunixin meglumine. In ischaemic-injured jejunum LPS permeability was significantly increased in horses treated with flunixin meglumine alone. Behavioural pain scores did not increase significantly after surgery in horses treated with flunixin meglumine. CONCLUSIONS: Treatment with systemic lidocaine ameliorated the inhibitory effects of flunixin meglumine on recovery of the mucosal barrier from ischaemic injury, when the 2 treatments were combined. The mechanism of lidocaine in improving mucosal repair has not yet been elucidated.     

肠相关性淋巴样组织研究概况

系统论述了肠相关淋巴组织的组成结构及其在肠道黏膜免疫中的功能作用，同时，概述了肠相关淋巴组织在消化道感染中的可能作用，并指出从黏膜免疫的角度来研究畜禽消化道疾病的病原体与消化道粘膜感染的关系是揭示这类疾病发生机理的一个新的突破点，它将为策划新的防制战略以控制畜禽消化道疾病的发生和消化道感染提供重要的依据。     

The timeline of lamellar basement membrane changes during equine laminitis development

Reasons for performing study: The timing of lamellar basement membrane (BM) changes occurring during laminitis development is incompletely understood. Objectives: To determine the temporal progression of lamellar BM changes and whether laminin‐332 (Ln‐332) γ2 cleavage products are generated during laminitis development. Methods: Eight clinically normal Standardbred horses were allocated into treatment (n = 5) or sham (n = 3) groups. The treatment group received, via nasogastric intubation, an oligofructose (OF) bolus (10 g/kg bwt) while the sham group was given water. Laminitis induction proceeded for 48 h followed by euthanasia. Lamellar biopsies were obtained prior to dosing and at intervals during the treatment period for analysis (at 12, 18, 24, 30 and 36 h and at 48 h following euthanasia). Results: Changes in lamellar collagen type IV and Ln‐332 were first observed at 12 h post dosing. A unique pattern of reactivity for the Ln‐332 γ2 antibody D4B5 occurred, in which reactivity was observed only in lamellar tissue affected by laminitis. No bioactive Ln‐332 γ2 proteolytic fragments were detected in lamellar samples. Conclusions: Basement membrane changes occurred early during the laminitis process. Direct Ln‐332 γ2 cleavage to release biologically active products did not appear to occur. Thus loss of stability or protein interaction of the BM is probably responsible for the γ2 specific reactivity observed. Potential relevance: Basement membrane changes may a first step in lamellar failure occurring prior to detection with conventional methods. Thus, more sensitive detection methods of BM changes are required to study laminitis development.     

Synovial membrane microarthroscopy of the equine midcarpal joint

OBJECTIVE: To evaluate the value of microarthroscopy in the equine midcarpal joint using the vital stains methylene blue, trypan blue, neutral red, and Janus green B to observe components of the synovial lamina propria, vascular architecture, and synoviocytes. STUDY DESIGN: Experimental. ANIMALS: Ten horses. METHODS: Microarthroscopy of left and right midcarpal joints was performed with and without vital staining of the synovium. Four vital stains (methylene blue, trypan blue, neutral red, and Janus green B) were evaluated, with each stain used in 5 joints. Synovial biopsy specimens were collected from the dorsomedial and dorsolateral aspects of the joint. RESULTS: All dyes were biocompatible. At x 60 without vital staining, synovial surface topography, vascular network, and translucency were observed. Intra-articular vital dyes improved evaluation of synovial surface topography. At x 150 with vital staining, individual synoviocytes were clearly identified with all dyes, except neutral red. Although methylene blue provided the best in vivo microscopic differentiation of the structure of the intima, trypan blue had superior retention in conventionally processed synovial biopsies. CONCLUSIONS: Methylene blue, trypan blue, neutral red, and Janus green B stains can be used safely for microarthroscopy. Good visualization of cells and vascular network can be obtained by microarthroscopy, and microarthroscopic evaluation of the synovium compares favorably with conventional histologic evaluation of biopsy specimens. CLINICAL RELEVANCE: Microarthroscopy may be beneficial in both research and clinical diagnosis of equine articular diseases.     

禽网状内皮组织增生病病毒感染对SPF雏鸡血液和局部淋巴组织CD4+/CD8+细胞及相关细胞因子表达的影响

旨在探讨禽网状内皮组织增生病病毒（reticuloendotheliosis virus,REV）对SPF雏鸡血液和局部淋巴组织中T淋巴细胞数量以及相关细胞因子表达的影响。将96只1日龄SPF雏鸡随机分为REV感染组和对照组,应用流式细胞术、酸性α-醋酸萘酯酶染色（acid α-naphthyl acetate esterase,ANAE）、荧光定量PCR等方法对上述指标进行检测。试验数据表明,与对照组相比,感染组雏鸡血液CD4⁺T淋巴细胞数量在第7~35天显著降低、CD8⁺T淋巴细胞数量在第7~28天显著升高,CD4⁺/CD8⁺比值在第7~28天显著降低（均P<0.05或P<0.01）;局部淋巴组织哈德尔腺（Hader’s gland,HG）、派伊尔结（Peyer’s patch,PP）和盲肠扁桃体（caecal tonsil,CT）中ANAE⁺T淋巴细胞数量均显著降低（均P<0.05或P<0.01）;GH、PP和CT中细胞因子IL-2、IFN-γ和TNF-α 转录量都有不同程度升高。本研究表明,REV感染引起雏鸡血液中CD4⁺ T淋巴细胞数量降低、CD4⁺/CD8⁺T淋巴细胞比例失衡、局部淋巴组织中T淋巴细胞数量相对减少及细胞因子TNF-α转录持续升高与REV造成感染雏鸡细胞免疫功能显著降低密切相关。     

Treatment of corneal ulceration and bullous keratopathy using a nictitating membrane flap in two horses

Objective

The aim of this study was to describe placement of a nictitating membrane flap as a treatment for corneal ulceration and bullous keratopathy in two horses.

Animals Studied

A 13-year-old American Saddlebred mare presented for severe corneal edema, superficial stromal ulceration, and a central bulla of the left eye. A 4-year-old Trakhener stallion also presented with a large axial bulla of the left eye with concurrent severe corneal edema and a deep stromal ulcer.

Procedure

A complete ophthalmic examination was performed. Samples were obtained for corneal cytology, and both horses were started on aggressive medical therapy. Both underwent general anesthesia for placement of a nictitating membrane flap and a subpalpebral lavage system (SPLS).

Results

Corneal cytology for each horse revealed a mixed bacterial population. Moderate Pseudomonas aeruginosa was cultured from the mare, while Aspergillus species and a few Enterococcus gallinarum were cultured from the stallion. The bullae in both horses resolved at 3 and 4 weeks and vision returned in the affected eye 4.5 and 3 months postoperatively at the last follow-up, respectively.

Conclusion

Aggressive medical management with concurrent placement of a nictitating membrane flap is effective to treat bullous keratopathy in two horses. The described treatments could be used to treat horses that develop severe or progressive bullous corneal lesions.     

Detection of calprotectin and its correlation to the accumulation of neutrophils within equine large colon during ischaemia and reperfusion

REASON FOR PERFORMING STUDY: The cytosolic protein complex, calprotectin, is abundant in neutrophils and could be used to improve the ability to localise and assess neutrophil infiltration in the equine intestine during ischaemia and reperfusion (I/R), but further study is required. OBJECTIVES: To assess the number of calprotectin-containing cells by immunohistochemistry in correlation with direct counting and scoring of neutrophils in the equine colon during I/R. METHODS: One and 2 h ischaemia of the left dorsal colon were induced, followed by 30 min reperfusion under general anaesthesia or by 18 h reperfusion after anaesthetic recovery. Biopsies were processed for light microscopy and stained with H/E for detection of neutrophils. To identify calprotectin-containing cells, immunohistochemistry was performed on formalin-fixed tissues with the murine MAC 387 antibody and a biotin-free peroxidase staining procedure. The number of neutrophils within submucosal venules and the colonic mucosa were calculated and compared with the number of calprotectin-positive cells. RESULTS: The number of calprotectin-positive cells within submucosal venules and within the colonic mucosa correlated significantly with the accumulation of neutrophils within the corresponding tissue segments. Within the submucosal venules, both calprotectin-positive cells and H/E-stained neutrophils increased with duration of ischaemia and peaked after 30 min of reperfusion. After 18 h reperfusion the number of these cells declined within the vessels. After 2 h ischaemia, neutrophils started to migrate into the mucosa towards the epithelium, with a significant increase over time during reperfusion, and peak infiltration after 18 h reperfusion. CONCLUSIONS: Neutrophil infiltration into the colon after I/R is a time-dependent process, involving migration through the submucosa towards the epithelium.     

State of the art: Stem cells in equine regenerative medicine

According to Greek mythology, Prometheus' liver grew back nightly after it was removed each day by an eagle as punishment for giving mankind fire. Hence, contrary to popular belief, the concept of tissue and organ regeneration is not new. In the early 20th century, cell culture and ex vivo organ preservation studies by Alexis Carrel, some with famed aviator Charles Lindbergh, established a foundation for much of modern regenerative medicine. While early beliefs and discoveries foreshadowed significant accomplishments in regenerative medicine, advances in knowledge within numerous scientific disciplines, as well as nano‐ and micromolecular level imaging and detection technologies, have contributed to explosive advances over the last 20 years. Virtually limitless preparations, combinations and applications of the 3 major components of regenerative medicine, namely cells, biomaterials and bioactive molecules, have created a new paradigm of future therapeutic options for most species. It is increasingly clear, however, that despite significant parallels among and within species, there is no ‘one‐size‐fits‐all’ regenerative therapy. Likewise, a panacea has yet to be discovered that completely reverses the consequences of time, trauma and disease. Nonetheless, there is no question that the promise and potential of regenerative medicine have forever altered medical practices. The horse is a relative newcomer to regenerative medicine applications, yet there is already a large body of work to incorporate novel regenerative therapies into standard care. This review focuses on the current state and potential future of stem cells in equine regenerative medicine.