Optimizing methods in immunocytochemistry: one laboratory's experience

Abstract: The addition of immunocytochemical staining procedures to a diagnostic cytology service enables greater specificity of interpretation for many common disease conditions, especially neoplastic diseases. However, well‐tested immunohistochemical techniques may require modification for cytologic specimens, and other considerations are necessary when working with air‐dried cells. In this article, we describe our experience in evaluating options for sample transport and handling, and discuss methods for obtaining control cells from a variety of tissues for use in immunocytochemical staining. Important immunocytochemical principles and techniques, including fixation, antigen retrieval, and use of primary and secondary antibodies in manual and automated staining systems are described as used in our laboratory for cytologic specimens. Although we emphasize methods relevant to diagnostic laboratories receiving samples from external clients, the information is also applicable to any laboratory interested in adding or enhancing immunocytochemical services.     

一例猫胫腓骨骨折的诊治

猫胫腓骨骨折发生后,由于猫胫腓骨很细,且近心端附着丰富的肌肉,而远心端很少,如果单纯采用外固定或内固定的治疗方法,效果往往不理想。文章就一例猫胫腓骨骨折采用髓内针和金属丝进行内固定后,又在外面用夹板进行外固定,术后恢复良好的病例来进行研究,希望能对猫胫腓骨骨折的治疗提供参考。     

草地生物固氮与集约化草地畜牧业

草地生物固氮不仅是草地生态系统氮素循环输入过程的重要部分，而且与集约化草地畜牧业的建立和退化草场的恢复有重要的关系。在草地生态系统生物固氮的３种方式中，豆科植物根瘤生固是重要方式，细菌自主固氮值得重视，而联合固氮在草地生物固氮中有着广泛的应用前景与潜力。     

Influence of ventral fixation techniques on atlantoaxial joint fusion in canine models with dens partial resection

We evaluated the completeness of bony fusion of the atlantoaxial joint (AAJ) through polymethylmethacrylate fixation (PMF) and atlantoaxial plate fixation (APF) using six canine models with dens partial resection. In both groups, the hydroxyapatite content at the AAJ was measured up to 7 months postoperatively using quantitative computed tomography. Histological assessment revealed fibrous fusion in the PMF group. Meanwhile, in the APF group, only one dog achieved fibrous fusion, whereas the remaining three showed bony fusion. To our knowledge, this study was the first to evaluate AAJ fusion histologically after PMF and APF. The present study demonstrates that PMF and APF may stabilize the AAJ without clinical complications. Therefore, PMF and APF are clinically useful fixation methods for atlantoaxial instability.     

微量补体结合试验检测牛、羊副结核病的研究

采用《中华人民共和国出入境检验检疫行业标准》“副结核病补体结合试验操作规程”方法(简称常量法)和微量补体结合试验(简称微量法)检测3677头进口牛羊的副结核病抗体并进行比较。结果表明，常量法和微量法的溶血素效价测定值相同，补体效价测定值近似，阴性血清对照、抗原对照、补体对照、红细胞对照、血清抗补体结合对照结果均相同，阳性对照血清效价微量法比常量法提高1个滴度，被检血清的阳性检出率微量法高于常量法。比较37℃水浴及37℃恒温培养箱环境下的微量法补体结合试验结果，溶血素效价、补体效价、各项对照试验结果完全相同，被检血清在37℃水浴反应更加充分。比较用变态试验和补体结合试验检验3677头牛羊副结核病的结果、以及用补体结合试验、变态反应和酶联免疫吸附试验方法同时检测2024头牛的结果，发现它们的相关性很小。     

Reactivity of workshop monoclonal antibodies on paraformaldehyde-fixed porcine blood mononuclear cells

One hundred sixty-four monoclonal antibodies (mAbs) of the second international swine CD workshop were tested for their reactivity with porcine blood mononuclear cells before and after fixing the cells with varying concentrations of paraformaldehyde (PFA) (1, 5 and 10 g l⁻¹). A total of 38 (out of 134) positive reacting mAbs were significantly affected in their binding behavior on fixed cells. Modulation was seen as reduction in binding (staining intensity and/or % positive cells, n=18) or in elevated values (n=20). Modified mAb binding occurred after fixing cells with 5 to 10 g l⁻¹ PFA.     

榆林沙区主要人工固沙灌丛营养成分及对放牧羊只生育影响的研究

人工固沙灌丛粗蛋白、粗脂肪平均含量比天然草地牧草分别高出１６．２２％、４３．５０％,钙的含量除紫穗槐外,平均高出２７．３７．％。磷的含量低于天然牧草,氨基酸总量平均值为１０．８５％,略低于草地牧草,７种必须氨基酸含量与天然牧草接近,沙柳、紫穗槐的镁、锌含量高,踏郎铁含量。铜的含量各灌丛均高。灌丛地放牧,杂交绵羊体重平均加３．４￣３２．０％,羊毛增加２０．０￣７９．３％,产羔率也有较大提高。     

Fixation of Cancellous Bone and Physeal Fractures in Dogs and Cats. A Comparison of the Use of Self-reinforced Biodegradable Devices to the Use of Metallic Devices and External Fixations

The use of self-reinforced biodegradable devices made of polyglycolide in the cancellous bone and physeal fractures of dogs and cats was compared to the use of metallic devices and external fixations on similar fractures. The series consisted of 64 dogs and 22 cats divided into 6 comparable groups. The patients in the 2 groups fixed with biodegradable devices started to use their operated limbs earlier than in the other groups. The same 2 groups also healed clinically (showed no lameness) earlier than the other groups. Radiographically there was no statistical difference between the 6 groups.On the basis of this study it may be concluded that the fixation with self-reinforced biodegradable devices is as suitable for the fixation of cancellous bone and physeal fractures of dogs and cats as the fixation with metallic devices or external fixation. This biodegradable technique has additional benefits allowing the patients to feel less pain in their operated limbs and making secondary operations to remove the devices unnecessary.     

不同苜蓿根瘤菌固氮能力和黄酮变异研究

从重庆、安徽和新疆3个地区的宿主苜蓿上分别分离纯化出1个根瘤菌菌株。将3个菌株分别接种到WL323和阿尔冈金两个品种苜蓿上。分析不同处理的固氮能力和它们的芦丁的含量，以及黄酮含量和固氮的关系。结果表明：固氮能力、芦丁、黄酮含量都受到根瘤菌菌株、苜蓿品种和两者的相互作用影响；并且各组合的芦丁含量与固氮量之间成负相关，黄酮含量越高，固氮量越少。     

弗兰克氏菌在农业生产中的应用

农业的可持续发展是现代农业追求的目标,而生物固氮被认为是最有效的方式,其中,弗兰克氏菌以其特有的固氮优势在农业生产应用中受到了越来越多的重视。介绍了弗兰克氏菌在农业生产应用中的现状、前景及存在的问题。     

Arthroscopic guided repair of a slab fracture of the fourth carpal bone in an 8-month-old thoroughbred horse

An 8-month-old thoroughbred colt presented with sudden onset right forelimb lameness. A radiographic series of the right carpus was performed, and it revealed a slab fracture of the fourth carpal bone and fracture of the proximal part of the third metacarpal bone. Arthroscopically guided repair of the slab fracture of the fourth carpal bone with a 3.5 mm cortex screw and lag screw fixation of the fracture of the proximal part of the third metacarpal bone were performed. The horse started to race at 32 months old and started in 65 races over three years without any trouble associated with the right carpus.     

土壤联合固氮菌的研究进展

面临人类社会普遍出现的粮食、人口、环境、能源、资源等问题的困扰,从可持续发展的角度出发,生物固氮逐渐引起了各国科学家的重视,测试技术的进步及细胞工程与基因工程等技术的出现使生物固氮的研究日趋深入。通过阐述生物固氮中细胞联合固氮的研究现状及其方法与手段,为与之相关的研究者提供了参考。     

狗牙根内生固氮菌生长条件

从已分离的多个狗牙根(Cynodon dactylon)内生固氮菌株中筛选到两株固氮酶活性较高的菌株7D和BM13,为明确固氮菌株对环境因子的适应性,研究了在不同的生长环境条件下菌株的生长量。采用Ashby无氮培养基,测定了温度、pH、渗透压以及不同氮源和碳源等环境条件对固氮菌生长的影响。结果表明,在10~40℃的温度范围内、pH值3.92~9.64的酸碱环境和30g·L~(-1) NaCl的高渗透压下,固氮菌7D和BM13均能正常生长;菌株能耐受0~10mmol·L~(-1)的NH_4~+和NO_3~-,而且能利用蔗糖、甘油、麦芽糖、甘露醇和葡萄糖等多种碳源生长。说明狗牙根内生固氮菌株7D和BM13对环境因子有很强的抗逆性和适应性,但两菌株之间存在一定差异,可望进一步研发成为优良的固氮微生物肥料生产菌种。     

固氮菌对燕麦不同生育期促生作用的研究

研究了固氮菌对兰州地区燕麦不同生育期的促生作用。结果表明,假单胞菌属Pseudomonas sp. N4,动胶杆菌属Zoogloea sp. W6,生脂固氮螺菌Azospirillus lipoferum C6,固氮菌属Azotobacter sp. O5和Azotobacter sp. W5对燕麦有促生作用。拔节期,接种固氮菌后燕麦株高增加0.73%~12.11%,叶绿素增加1.14%~39.43%,地上植物量增加21.45%~43.55%,地下植物量增加51.85%~130.86%,根冠比增加7.7%~76.9%,总植物量增加2.00%~45.36%,粗蛋白增加3.02%~25.57%;半量化肥 固氮菌处理的株高增加1.96%~5.82%,叶绿素和地上植物量没有增加,地下植物量增加3.47%~33.17%,根冠比增加15.38%~30.76%。成熟期,各个处理促生作用降低,处理间差距缩小,Pseudomonas sp. N4和Azotobacter sp. O5促生效果好。     

Bilateral correction of metatarsal rotation in a dog using circular external skeletal fixation

An 8-month-old castrated male Saint Bernard was evaluated for bilateral hind limb lameness. Lameness was ascribed to bilateral metatarsal rotational abnormalities on the basis of the physical examination and radiographic evaluations. Staged, bilateral deformity correction and tarsometatarsal arthrodeses were performed using circular external skeletal fixators. The dog's gait improved following surgery and the fixators were removed 3 (left hind paw) and 4 (right hind paw) months following surgery. Twenty months after the initial surgery, the owner reported that the dog was walking well without apparent lameness. Previous reports suggest that metatarsal rotation is untreatable; however, our results suggest that surgical correction of this deformity can substantially improve limb function in dogs affected with metatarsal rotation.     

青海豆科植物根瘤及固氮特性的研究

对青海野生及栽培豆科植物结瘤以及固氮状况进行了调查研究,栽培豆科植物均能自然结瘤,野生豆科植物结瘤率近为45%。青海地区豆科植物根瘤大多圆形,棒状或指状,形状较为规则。栽培豆科植物根瘤多数为粉红色,而野生豆科植物多数为白色,棕色或黄色。125种根瘤样品用ARA测定结果表明,30%根瘤为无效根瘤,不同豆科植物不同地区根瘤固氮活性相差较大,一般固氮活力都较低,乙炔还原活性小于1umolC2H2/gFw.h.的检测样品有40%,最高达30umolC2H2/gFw.h,根瘤固氮活性与生育期,生活习性及生态条件有关。     

Two techniques for supplementing interlocking nail repair of fractures of the humerus, femur, and tibia: results in 12 dogs and cats

OBJECTIVE: To describe 2 devices for improving stabilization of inadequately stabilized interlocking nail (ILN) repairs of the humerus, tibia, and femur in dogs and cats. STUDY DESIGN: Prospective study. ANIMALS: Twelve client-owned dogs and cats. METHODS: Two devices to further stabilize ILN repair of inadequately stabilized diaphyseal fractures were developed. Device 1 was an axial extension for the ILN that was connected to a conventional type I external skeletal fixator (ESF) with a short connecting bar. Device 2 had hybrid ILN bolt/ESF pins that were used to lock the ILN and serve as the pins for a type I ESF. Devices were used at the initial surgery when the stability of ILN repair was considered inadequate based on palpable fracture segment movement, insufficient medullary canal filling of the ILN at the fracture site, or when the ILN was used in a buttress mode. Outcome was obtained by recheck examinations, radiography, and telephone interview. RESULTS: Device 1 was applicable to fractures of the humerus and femur, but was not used for fractures of the tibia because the ILN extension would have interfered with the stifle. No gross loosening of the ILN/ESF extension connection to the ILN occurred. Device 2 was easily placed and used in the humerus, femur, and tibia. Device 2 allowed removal of the ILN interlock to one or both main fracture segments non-invasively. Clinically, both devices added stability compared with ILN repair alone. Both devices facilitated controlled destabilization of the fracture repair as healing progressed. Complications of pin tract infection, and premature hybrid bolt/ESF pin loosening resulting in premature ESF removal each occurred in 1 patient. Four of 28 hybrid ILN/ESF pins were grossly loose at 4- or 6-week postoperative recheck examinations. Outcomes were excellent (9), good (1), fair (1), and poor (1). CONCLUSIONS: Inadequately stabilized ILN repair of fractures can be stabilized by use of either device, both of which also permit controlled destabilization of the repair during healing. Device 2 can be used when non-invasive removal of the ILN interlock is desired during healing. CLINICAL RELEVANCE: These 2 devices should be considered as alternative methods for stabilization of inadequately stabilized ILN repairs in dogs and cats, or when controlled destabilization of an ILN fracture repair is desired.     

Surgical correction of severe strabismus and enophthalmos secondary to zygomatic arch fracture in a dog

A grossly displaced segmental zygomatic arch fracture with marked ventro-lateral deviation of the left globe was diagnosed in a 3-month-old male German Shepherd dog following a bite injury. The fracture was approached via a modified lateral orbitotomy and a fragment of the lacrimal bone removed. The rostral portion of the fracture was stabilized with a 5-hole 2.0 dynamic compression plate bone plate. The surgical correction achieved sufficient skeletal fixation for proper anatomical reduction of the globe and excellent cosmetic and functional outcomes.     

Surgical Repair of a Pelvic Fracture in a Ferret (Mustela putorius furo)

A 9-month-old, chemically spayed female ferret (Mustela putorius furo) presented after a fall (5.5 m) from the first floor of an apartment building. The animal was nonambulatory, with pain localized in the pelvic region. Radiographic examination revealed a bilateral fracture of the iliac body with medial displacement of the left hemipelvis and narrowing of the pelvic canal. Rib fractures, pulmonary contusions, and pleural effusion were also present on thoracic radiographic images. After emergency stabilization of the patient, the fracture of the left iliac body was successfully repaired using a composite internal fixation system with K-wires and polymethylmethacrylate. The implants allowed fracture healing despite loosening of one of the K-wires. Bony healing was achieved within 25 days, including the restoration of pelvic rigidity and maintenance of the diameter of the pelvic canal. Orthopedic examination was unremarkable 4 months following the surgical procedure. To the authors’ knowledge, this is the first described open reduction of a pelvic fracture in a ferret.     

牧草联合固氮研究概况

联合固氮是草原土壤氮素的重要来源,但是目前对于牧草联合固氮方面的研究却仍然比较滞后。综述了牧草联合固氮菌的种类、固氮酶活性、对牧草的促生作用、影响固氮的因素等方面的研究概况,以期能够引起人们对牧草联合固氮菌的重视。