Comparison of ultrasound‐guided vs. ‘blind’ techniques for intra‐synovial injections of the shoulder area in horses: Scapulohumeral joint,bicipital and infraspinatus bursae

Reasons for performing study: Diagnosis and treatment of pathologies involving synovial structures in the shoulder region are technically difficult. Ultrasound‐guided (UG) injection techniques have been shown to be highly accurate, safe and reliable for various structures in human and equine patients. Objectives: To develop easy‐to‐use and reliable UG injection techniques for the infraspinatus bursa (IB), bicipital bursa (BB) and scapulohumeral joint (SHJ) and to compare them with conventional ‘blind’ methods. Methods: Eight pairs of equine shoulders were positioned on a computed‐tomography (CT) table with left and right shoulders randomly assigned to Operator A (UG) and Operator B (‘blind’). Contrast medium mixed with methylene blue was injected into the target structures. Time and number of attempts for correct needle placement and ease of injection were recorded. A CT scan of each shoulder was performed after every injection to evaluate the location of contrast material. Once injections and CT scans were completed, anatomic dissections were conducted. Statistical analysis was used to compare UG and ‘blind’ techniques with significance set at P<0.05. Results: Intra‐synovial injections using UG techniques were successful in all 24 synovial structures compared with 14/24 with conventional methods. The median number and range of needle repositioning to obtain accurate needle placement was 1 (1–2) for UG and 2 (1–4) for the ‘blind’ techniques. The median time and range required for accurate needle placement was 75.5 s (32–210 s) for UG and 43.5 s (11–140) for ‘blind’ technique. In 31% of all attempts, ease of injection was not indicative of successful intra‐synovial administration. Conclusions: Ultrasound‐guided injections of IB, BB and SHJ proved to be highly reliable and more accurate than conventional ‘blind’ techniques. Potential relevance: Ultrasound‐guided injection techniques may be of great help for equine practitioners in the diagnosis and treatment of pathologies involving synovial structures in the shoulder region. Further work is needed to confirm these results in clinical cases.     

Determination of MMP-2 and -9 activities in synovial fluid of horses with osteoarthritic and arthritic joint diseases using gelatin zymography and immunocapture activity assays

REASON FOR PERFORMING STUDY: Matrix metalloproteinases (MMPs)-2 and -9 activities have been found elevated in synovial fluid from various joint diseases in man. However, in the horse few data are available. OBJECTIVES: To explore the clinical significance of MMP-2 and -9 activities in synovial fluid of horses with different forms of joint diseases. METHODS: Gelatin zymography and MMP-2 and -9 immunocapture activity assays were applied on synovial fluids from control joints and joints with aseptic joint disease (AJD) and septic arthritis (SA). Additionally, MMP-2 and -9 activities were measured in samples from SA to monitor the disease process. RESULTS: Zymographic analysis revealed that samples from AJD and SA contained significantly increased latent MMP-2 activity compared to controls. Samples from SA showed significantly increased monomeric latent MMP-9 activity compared with all other affected joints and controls. Trace amounts of MMP-9 activity, due to the active and dimer form, were detected in samples from SA; however, these bands were absent in samples from AJD and controls. Using immunocapture activity assays, MMP-2 and -9 activities were found to be significantly elevated in joints from SA compared to controls and AJD samples. MMP-2 activity in samples from AJD was significantly increased compared to controls. Both MMP activities decreased in the joints from SA in the course of successful therapy. CONCLUSIONS: Data from zymographic analysis confirmed that MMP-2 and -9 were elevated in equine joint diseases. Immunocapture activity assays have been shown to be suitable for the quantitative determination of MMP-2 and -9 activities in synovial fluid of horses. Both MMP-2 and -9 activities seem to be useful to indicate SA, and MMP-2 activity might be a suitable marker for AJD. POTENTIAL RELEVANCE: These findings encourage the potential use of MMP-2 and -9 as additional aids to clinical investigation. Further work is required to validate the clinical significance of MMP activities in the progress of different joint diseases in horses.     

Cartilage-derived retinoic acid-sensitive protein in equine synovial fluid from healthy and diseased joints

Reasons for performing study: More sensitive and specific diagnostic methods for early detection of changes in the joint cartilage are needed. Cartilage‐derived retinoic acid‐sensitive protein (CD‐RAP) is a potential marker of cartilage synthesis and regeneration. This is the first study on equine CD‐RAP. Objectives: To evaluate the ability of a commercially available human sandwich ELISA assay to detect equine CD‐RAP in synovial fluid from healthy and diseased joints. Methods: Synovial fluid was collected from 28 horses with no signs of joint disease and from 5 with induced inflammatory arthritis. CD‐RAP concentrations were measured using a human CD‐RAP ELISA. Intra‐ and interassay imprecision of the assay were evaluated by multiple measurements on pools of equine synovial fluid. Assay inaccuracy was determined by linearity under dilution. Results: The assay showed moderate to large intra‐ and interassay variation when applied to equine synovial fluid. Equine CD‐RAP was detected in synovial fluid from healthy horses ranged at 8.2–52 ng/ml. Repeated arthrocentesis (after injection of isotonic saline), age, joint or gender did not significantly affect CD‐RAP concentrations. Twelve hours after intra‐articular injection of lipopolysaccharide, concentrations of CD‐RAP were significantly lower than after injection of isotonic saline and remained significantly lower until the end of the study at 144 h. Conclusion and potential relevance: The assay is suitable for longitudinal monitoring of CD‐RAP concentration in individual horses. Disease significantly influenced CD‐RAP levels. Similar to previous results obtained in man, CD‐RAP seems to be a marker of cartilage synthesis and/or regeneration in horses.     

奶牛正常腕、跗关节滑液细胞学的研究

实验对6头健康有牛的腕关节、跗关节滑液的细胞学成分进行了研究,确立了关节滑液细胞的计数方法。用瑞—姬氏混合染色发现奶牛关节滑液细胞分为4类,即:淋巴细胞、嗜中性白细胞、巨噬细胞和退化细胞。滑液中嗜中性白细胞极显著低于血液中的嗜中性白细胞。未见嗜酸性粒细胞和嗜碱性粒细胞。     

MAGNETIC RESONANCE IMAGING FINDINGS IN HORSES WITH SEPTIC ARTHRITIS

Fourteen horses with septic arthritis underwent high‐field (1.5 T) magnetic resonance imaging (MRI). Septic arthritis was diagnosed based on results from historical and clinical findings, synovial fluid analyses and culture, and radiographic, ultrasonographic, arthroscopic, and histopathologic findings. MR findings included diffuse hyperintensity within bone and extracapsular tissue on fat‐suppressed images in 14/14 horses (100%), joint effusion, synovial proliferation, and capsular thickening in 13/14 horses (93%), bone sclerosis in 11/14 horses (79%), and evidence of cartilage and subchondral bone damage in 8/14 horses (57%). Intravenous gadolinium was administered to five of the 14 horses and fibrin deposition was noted in all horses. Other findings after gadolinium administration included synovial enhancement in 4/5 (80%) horses, and bone enhancement in 1/5 (20%) horses. The MR findings of septic arthritis in horses were consistent with those reported in people. MRI may allow earlier and more accurate diagnosis of septic arthritis in horses as compared with other imaging modalities, especially when the clinical diagnosis is challenging. It also provides additional information not afforded by other methods that may influence and enhance treatment.     

Biomarkers for identifying the early phases of osteoarthritis secondary to medial patellar luxation in dogs

The levels of tartrate resistant acid phosphatase (TRAP), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in synovial fluid (SF) and serum in cases of canine osteoarthritis (OA) were measured. OA was induced by a surgically-created medial patellar luxation in the left stifle of 24 dogs. SF and blood samples were collected at 1.5- and 3-month intervals, respectively. Every 3 months, one dog was euthanatized to collect tissue samples from both stifles. TRAP levels in SF and serum were measured using a spectrophotometer, and TRAP-positive cells in joint tissues were identified by enzyme histochemistry. MMP-2 and TIMP-2 in SF and serum were detected by Western blotting and ELISA, respectively. TRAP in SF from the stifles and serum was significantly increased (p < 0.05) after 3 months. TIMP-2 in SF and serum was significantly decreased (p < 0.05), whereas MMP-2 in SF was significantly increased (p < 0.05) during the progression of OA. Histochemistry revealed an increased number of TRAP-positive cells in tissues from OA-affected joints. Assays measuring TRAP, MMP-2, and TIMP-2 in SF and serum, and methods that detect increased numbers of TRAP-positive cells in the joint tissues can play an important role in identifying the early phases of degenerative changes in canine joint components.     

马关节滑液、血清乳酸脱氢酶及其同工酶和它们在疾病状态下的变化

用king氏法和聚丙烯酰胺凝胶电泳法测得马关节滑液和血清乳酸脱氢酶(LDH)及其同工酶正常值。滑液和血清5种LDH同工酶电泳迁移率相同。各种LDH同工酶的分子量和热稳定性均有明显差异。抗尿素作用,血清和滑液LDH同工酶基本相似。Mg~(++)是LDH激活剂,最佳激活浓度为10~(-3)M;Zn~(++)、Ca~(++)、Cu~(++)、Hg~(++)、谷胱甘肽、半胱氨酸、碘乙酸、EDTA等为LDH抑制剂。在四肢的急性或亚急性关节和骨疾病时,滑液LDH明显升高,其同工酶谱随疾病发生部位不同而不同,并与血清一致,但慢性关节疾病时LDH及其同工酶变化则不一致。     

Autologous processed plasma: cytokine profile and effects upon injection into healthy equine joints

This experimental controlled study was performed to evaluate the composition of autologous processed plasma (APP), and the effects of APP intra-articular injection into healthy equine metacarpophalangeal joints. The effects on joints were analysed with a short-phase protocol and a prolonged-phase protocol using saline-injected joints as controls. For the short protocol, horses received one intra-articular APP injection. Synovial fluid samples were collected prior to the injection and 3, 6, 24, 48, and 16 h after treatment. For the prolonged protocol, the joints received three weekly injections of APP, and samples were collected at 0, 7, 14, 21, and 28 days before APP administration. IL1-ra level was found to be increased in APP compared to plasma. Upon intra-articular administration of APP, transient (up to 24 h) increases in white blood cell (WBC) counts along with elevated protein and prostaglandin E₂ (PGE₂) concentrations were observed in the treated joints. Over the 28-day observation period, APP did not elicit changes relative to baseline levels, but WBC counts, PGE₂ and chondroitin sulphate concentrations were lower than those found in the control. In conclusion, APP intra-articular injection induced a mild and transitory inflammatory response but no inflammation reaction was observed over a longer period of treatment and observation.     

肝胆管盆空肠Roux-y定向吻合术治疗83例肝胆管结石的疗效观察

采用肝内1～3级胆管切开取石,肝胆管盆空肠Roux-y定向吻合术治疗肝内胆管结石92例。手术切开胆总管后,采用边夹、边切、边缝、边牵引方法逐渐向上显露和切开肝内1～3级胆管,取净结石,解除肝胆管狭窄,修整肝胆管成一个宽大、平整光滑的肝胆管盆;然后行肝胆管盆空肠Roux-y定向吻合术、并建立一个联合的抗返流装置,术后以胆系及溃疡病症状为内容随诊83例,随诊时间3个月～4年。结果疗效优者44例,良者34例,差者5例,优良率约达94％。92例术后均行胆道引流管造影,发现9例肝内胆管终末支有残石,残石率为9.8％。讨论了不横断胆总管的肝胆管盆Roux-y定向引流术的优越性,认为该术式既能取净结石及解除肝内胆管狭窄,又能抗食糜逆流和可避免术后并发消化性溃疡,因此是治疗肝内胆管结石比较理想和有效的术式。