OPG/RANKL/RANK系统的生理功能

骨是一个代谢活跃的器官,在整个生命周期中都在进行不断的更新与重建。在骨形态发生和重建的过程中主要受到细胞因子和激素的调节,同时骨代谢也受到OPG/RANKL/RANK系统的调控。论文主要对骨骼生理结构与骨组织细胞之间的关系,骨代谢和骨的形成过程,OPG/RANKL/RANK系统的生理功能,OPG/RANKL/RANK系统的影响因子以及骨代谢紊乱所引起的疾病几个方面作一综述。     

Evaluation of bone healing in canine tibial defects filled with cortical autograft,commercial-DBM,calf fetal DBM,omentum and omentum-calf fetal DBM

The present study was conducted to compare the effects of xenogenic bovine fetal demineralized bone matrix (DBM), commercial DBM, omentum, omentum-calf fetal DBM, cortical autograft and xenogenic cartilage powder on the healing of tibial defects in a dog model to determine the best material for bone healing. Seven male adult mongrel dogs, weighing 26.2 ± 2.5 kg, were used in this study. Seven holes with a diameter of 4-mm were created and then filled with several biomaterials. Radiographs were taken postoperatively on day 1 and weeks 2, 4, 6, 8. The operated tibias were removed on the 56th postoperative day and histopathologically evaluated. On postoperative days 14, 42 and 56, the lesions of the control group were significantly inferior to those in the other group (p < 0.05). On the 28th postoperative day, the autograft group was significantly superior to the control and omentum groups (p < 0.05). Moreover, calf fetal DBM was significantly superior to the control group. There was no significant difference between the histopathological sections of all groups. Overall, the omentum and omentum-DBM groups were superior to the control group, but inferior to the autograft, commercial-DBM, calf fetal DBM and calf fetal cartilage groups.     

甲状旁腺激素对骨代谢相关基因调控的研究进展

骨代谢包括骨形成和骨吸收两个过程。甲状旁腺激素是骨形成和骨重建过程中一个重要的调节因子,甲状旁腺激素通过G蛋白偶联受体、甲状旁腺激素/甲状旁腺激素相关肽受体的活动调节骨代谢。近年来甲状旁腺激素显著的临床效果,为骨骼合成代谢药物在骨质疏松症的治疗上开辟了一个新时代。作者主要综述了甲状旁腺激素在骨代谢中的作用机制及相关调控基因的作用。     

Relationship between latency of brainstem auditory‐evoked potentials and head size in dogs

Summary

Cranium and brainstem dimensions were measured in 32 postmortem dog heads. Positive correlations were found between cranium length (CL) and brainstem length (BL) (r=0.87), between cranium width (CW) and brainstem width (BW) (r=0.83), and between cranium distance (CD = CL CW/2) and brainstem distance (BD = BL+BW/2) (r=0.91). Positive correlation coefficients were also found between CL and CW (r=0.90), and between BL and BW (r=0.85). It was concluded that head size accurately reflected brainstem size. A least squares estimation of the brainstem distance (BD) from CL and CW values was BD = 10.9 + 0.16 (CL CW/2) (BD, CL and CW in mm).

Brainstem auditory evoked potentials (BAEPs) and cranium dimensions were measured in 43 dogs (86 ears) with different head size, body size, sex and age. Wave form, absolute and interpeak latencies and correlation coefficients, relating latencies to cranium dimensions and body weight, were analysed CL, CW, and CD were positively correlated with body weight (r=0.93, 0.70 and 0.93, respectively), and CL, CW, and CD were correlated with age (r=0.33, 0.52 and 0.40, respectively). BAEPs consisted of five distinct positive peaks (I to V). Secondary positive peaks following peaks I and II were seen in 60% (I') and 90% (II') of the recordings. Late waves were recorded in 90% (VI), 50% (VII), and 25% (VIII) of the recordings. Latencies increased with decreasing stimulus intensity level (from 90 dB to 10 dB hearing level, HL),especially for peaks I, II, V, and the I‐V interpeak interval Absolute and interpeak latencies were positively correlated with cranium distance and body weight. Correlation coefficients increased as wave latencies increased At 90 dB HL, the highest correlation coefficients, relating cranium distance to peak V and the I‐V interpeak latency, were 0.55 and 0.53 (P < 0.00001), respectively. Regression analysis showed that each 1 cm increase in cranium distance was accompanied by an increase of 0.006 ms in the latency of wave I, 0.03 ms for wave III, 0.05 ms for wave V, and 0.05 ms for the I‐V interpeak interval Regression analysis showed that an increase of 1 kg in body weight was accompanied by an increase of 0.001 ms in the latency of wave I, 0.005 ms for wave III, 0.011 ms for wave V, and 0.01 ms for the I‐V interpeak interval. It is concluded that head size, which accurately reflects brain size, is a relevant source (25%) of intersubject variance of BAEP latencies in the dog.     

Autologous point‐of‐care cellular therapies variably induce equine mesenchymal stem cell migration,proliferation and cytokine expression

Reasons for performing study: Autologous cellular therapy products including adipose‐derived stromal vascular fraction (SVF), bone marrow mononuclear cells (BMMNs), cord blood mononuclear cells (CBMNs) and platelet rich plasma are options for treatment of acute orthopaedic lesions while mesenchymal stem cells (MSCs) are culture expanded. These products may contribute to healing by secreting matrix proteins or growth factors, but they may also act on endogenous MSCs to facilitate healing. Objectives: To determine the effects of cell therapy products on MSCs function in vitro. The hypothesis was that cell therapy products promote MSCs functions including proliferation, migration and mediator release. Methods: Fat, bone marrow (BM), cord blood and platelets were obtained from 6 Quarter Horses. The BM‐MSCs and their autologous cell therapy products were co‐incubated in transwells. Mesenchymal stem cells proliferation, migration, gene expression and cytokine concentrations were determined. Results: All cell therapy products increased MSCs proliferation, but SVF induced significantly more proliferation than any other product. Also SVF elicited more MSCs chemotaxis and, along with BMMNs, significantly more MSCs chemoinvasion. Cord blood mononuclear cells stimulated MSCs to produce high concentrations of interleukin‐6 (IL‐6), transforming growth factor‐β1 (TGF‐β1), and prostaglandin E₂ (PGE₂). Stromal vascular fraction and platelet lysate did not stimulate MSCs but SVF and platelet lysate themselves contained high concentrations of PGE₂ and IL‐6 (SVF) and TGF‐β1 (platelet lysate). Conclusions: Autologous cell products variably stimulate MSCs functions with 2 primary patterns apparent. Products either contained preformed mediators that may have intrinsic healing function, or products stimulated MSCs to secrete mediators. Potential relevance: The specific clinical indications for these products may differ to include administration as a sole treatment modality prior to MSCs injection for intrinsic cell and cytokine activity (i.e. SVF) or administration concurrently with MSCs to activate MSCs for treatment of chronic lesions (i.e. CBMNs).     

A Therapeutic Potential for Marine Skeletal Proteins in Bone Regeneration

A vital ingredient for engineering bone tissue, in the culture dish, is the use of recombinant matrix and growth proteins to help accelerate the growth of cultivated tissues into clinically acceptable quantities. The skeletal organic matrices of calcifying marine invertebrates are an untouched potential source of such growth inducing proteins. They have the advantage of being ready-made and retain the native state of the original protein. Striking evidence shows that skeleton building bone morphogenic protein-2/4 (BMP) and transforming growth factor beta (TGF-β) exist within various marine invertebrates such as, corals. Best practice mariculture and the latest innovations in long-term marine invertebrate cell cultivation can be implemented to ensure that these proteins are produced sustainably and supplied continuously. This also guarantees that coral reef habitats are not damaged during the collection of specimens. Potential proteins for bone repair, either extracted from the skeleton or derived from cultivated tissues, can be identified, evaluated and retrieved using chromatography, cell assays and proteomic methods. Due to the current evidence for bone matrix protein analogues in marine invertebrates, together with the methods established for their production and retrieval there is a genuine prospect that they can be used to regenerate living bone for potential clinical use.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 8. Quantitative back-scattered electron scanning electron microscopy and confocal fluorescence microscopy of the epiphysis of the third metacarpal bone

AIM: To characterise and explain the increase in density evident by computerised tomography (CT) and radiography in companion studies as a response to training, in bone in the palmar and dorsal regions of the condyles of the third metacarpal bone (Mc3) of 2-year-old Thoroughbred horses.

METHODS: Compositional back-scattered electron (BSE) imaging in scanning electron microscopy (SEM) and confocal scanning laser microscopy (CSLM) were conducted on polymethyl methacrylate (PMMA)-embedded mediolateral slices of the right distal Mc3 from seven 2-year-old Thoroughbred horses trained on a racetrack and seven untrained horses kept at pasture. One left Mc3 from each group was studied in transverse section planes. This study focussed on regions of Mc3 found to differ in density between the trained and untrained horses in companion studies using CT and radiography.

RESULTS: The increase of bone density in the condyles of Mc3 in trained horses compared with untrained horses occurred, without prior osteoclastic resorption, via the deposition of new bone on pre-existing internal surfaces. Within prior marrow spaces of cancellous bone, there was also rapid formation of immature strands and fronds of bone which were more cellular and mineralised, and more lamellar bone tissue was deposited on these new scaffolding elements in the trained horses. Both resulted in increased bone volume fraction (BVF). The microscopic mineralisation density of the bulk of the new tissue was lower than in pre-existing bone, and CT and radiography underestimated the increase in BVF. The new tissue was thus probably less stiff at the microscopic scale than pre-existing bone, though its addition would stiffen the global structure.

CONCLUSIONS: In Mc3 of all the trained horses, there were obvious differences in microscopic structure compared with those from the untrained horses. Moderate, industry-standard levels of exercise used to prepare young horses for racing induced the formation of new bone in non-bone spaces in bone tissue, such that the bone organ should better withstand later increased levels of exercise.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 1. Study design,and clinical,nutritional, radiological and histological observations

AIMS: This is the first in a series of papers reporting studies in 2-year-old Thoroughbred racehorses that aimed to determine the response of musculoskeletal tissues to early training on grass and sand racetracks. In this paper, the experimental set-up of the whole study is described, and nutritional, workload, and clinical, radiographic and pathological outcomes are reported, including semi-quantitative assessment of macroscopic changes in articular cartilage.

METHODS: The study group comprised 14 two-year-old Thoroughbred fillies reared entirely at pasture. Of these, seven were selected by a licensed racehorse trainer to undergo a 4-week period of initial training in which they were taught to accept saddle and rider, followed by a 13-week period of flatrace training at a racetrack (Weeks 1–13); the other seven fillies were confined to large grass enclosures and were not trained. Nutrient, including macro- and trace-element intakes were estimated. Distances cantered or galloped and average velocities were quantified for the trained horses. All horses were observed daily, weighed approximately weekly, and underwent a clinical lameness examination at Weeks 5, 9 and 13. Distal forelimbs were radiographed prior to Week 1, during Weeks 7–8, and again at the end of the study, when macroscopic changes in articular cartilage of the proximal surface of the proximal phalanx were also scored after staining with India ink.

RESULTS: Dietary intakes met or exceeded recommended requirements for all nutrients except sodium, which was low in the trained horses. Bodyweight increased throughout the study in the untrained horses, and increased until Week 7 and then decreased slightly in the trained horses. Mean velocity data were used to define three stages of the training programme: Stage 1 comprised canter in Weeks 1–4; Stage 2 comprised canter in Weeks 5–8; and Stage 3 comprised canter in Weeks 9–13 and galloping twice weekly. Four of seven horses completed training. These covered a mean distance of 179.2 km at mean velocities (excluding gallops) of 7.63 m/sec (SD 0.58), 8.99 m/sec (SD 0.56), and 8.43 m/sec (SD 0.74) for Stages 1–3, respectively, and galloped 4.45 km at 14.4 (SD 0.1) m/sec. The three horses that did not complete training became lame in Weeks 9, 10 and 11, and covered 147.9 km at velocities of 7.38 m/sec (SD 0.44), 8.88 m/sec (SD 0.33) and 8.43 m/sec (SD 0.59) and galloped 2.1 km. Overall, slight or intermittent lameness in trained horses was noted on 76/655 (12%) of horse observation days. Swelling was evident on 284/655 (41%) of horse observation days in the metacarpophalangeal (MCPJ) and metatarsophalangeal (MTPJ) joints (92%), palmar metacarpal tendon region (7%) or carpus (1%). Swelling of the MCPJ or MTPJ was not associated with obvious lameness. Radiographic changes were minor and no gross lesions in bone or tendon tissue were evident except for one case of dorsal metacarpal disease. Post mortem, the cartilage of some MCPJ and MTPJ had obvious wear lines and high lesion scores, which were not consistently related to clinical evidence of pain, lameness or joint swelling. Mean lesion scores were not significantly different between the MCPJ and MTPJ, or between trained and untrained horses.

CONCLUSIONS: Workload can be readily quantified in racehorses under semi-commercial training conditions. Obvious lesions in cartilage of the MCPJ or MTPJ were present in some trained and some untrained horses and not consistently associated with clinical evidence of lameness, joint swelling or change in other connective tissues.

CLINICAL RELEVANCE: Workload data in racing horses are likely to be highly relevant for studying the pathogenesis of changes in bone, tendon and cartilage during training, for training management and for risk analysis in racehorse populations. Although obvious cartilage lesions produced little clinical effect, such lesions have previously been shown to be progressive and to prejudice athletic capability. Detection of such occult lesions in young horses will require more sophisticated detection methods.     

高压/酶解/闪式高速提取对金华火腿骨呈味物质的影响

以金华火腿骨为研究对象,研究不同提取方法对其呈味物质释放的影响。通过感官评价及可溶性糖、有机酸、5′-核苷酸和游离氨基酸的测定分析不同处理组间样品的差异,同时采用偏最小二乘回归法（partial least squares regression,PLSR）对提取物感官和呈味物质间进行相关性分析。感官评价发现,高压蒸煮的样品的鲜味、咸味和可接受度最高,其协同滋味的综合评分最佳;呈味物质分析发现,原液中所有呈味物质含量都最低,高压-复配酶解处理的样品中可溶性糖总量最高,高达132.68 mg/100 g;经过酶解处理样品有机酸含量显著增加（P<0.05）,达到3 733.32 mg/100 g;样品经不同处理后,5′-核苷酸含量呈现显著性增加（P<0.05）,总含量最高的是经过高压蒸煮处理的样品,质量分数高达1.24 mg/100 g,是原液和其他处理组样品的2.38～12.4倍;与样品原液相比,游离氨基酸总量都显著增加（P<0.05）,高压蒸煮处理的样品中含量最高,总质量分数为642.44 mg/100 g。借助偏最小二乘回归方法对样品的感官属性及呈味成分进行相关性分析,发现样品间具有显著性差异（P<0.05）,不同的处理方式所释放的呈味物质分布规律显著不同,高压蒸煮的样品与咸味和鲜味具有显著相关性（P<0.05）;酶解处理的样品与酸味和甜味有较强相关性,其他组样品与感官与呈味属性相关性不强。结论：不同处理方式的金华火腿骨具有不同的风味,其风味特征与多种物质有关,该研究结果为火腿骨的深度开发提供了理论指导,提高金华火腿骨的附加值。