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).     

桦褐孔菌多糖对急性感染弓形虫小鼠c-Jun、IL-12和IL-1β的影响研究

为了探讨桦褐孔菌多糖对急性感染弓形虫小鼠核转录因子c-Jun、IL-1β和IL-12的影响,本试验在建立急性感染弓形虫小鼠模型后,随机将其分成3组:阴性组、急性感染弓形虫小鼠模型组和桦褐孔菌多糖试验组。采用RT-PCR法检测肝组织中c-Jun及脾组织中IL-1β和IL-12的mRNA表达;Western blot法检测c-Jun蛋白的表达。结果显示:c-Jun、IL-1β和IL-12的mRNA分别在360、736和978 bp处扩增出单一电泳条带,且c-Jun蛋白在39 ku处表达,均呈现出弓形虫感染组表达量最高,桦褐孔菌多糖试验组次之,对照组最弱。说明桦褐孔菌多糖可能通过下调c-Jun过度表达,抑制IL-1β、IL-12的过度表达,部分阻抑了AP-1信号通路转导,从而减轻炎症反应,达到抗弓形虫感染的目的。     

栀子苷对酵母多糖致大鼠肠黏膜屏障损害作用的研究

目的 探讨栀子苷对酵母多糖（zymosan,Zym）致大鼠肠黏膜屏障损害的作用。方法 通过腹腔注射Zym-石蜡混悬液建立大鼠肠黏膜屏障损害模型,将模型大鼠随机分为模型组、栀子苷组、乌司他丁组,腹腔注射Zym后即刻干预,12 h后重复干预1次。另设正常对照组。造模24 h后测定大鼠血清肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、白介素-1β（interleukin-1β,IL-1β）、白介素-6（interleukin-6,IL-6）、白介素-10（interleukin-10,IL-10）、D-乳酸（D-lactate,D-LA）、二胺氧化酶（diamine oxidase,DAO）、内毒素（endotoxin,ET）水平,并取小肠进行病理组织学检查。结果 模型组大鼠血清TNF-α、IL-1β、IL-6、IL-10、D-LA、DAO、ET水平较正常对照组显著增高（P<0.01）;病理组织学检查可见黏膜上皮水肿,黏膜上皮细胞排列紊乱,黏膜上皮炎症细胞增多,固有层松散、充血;与模型组比较,栀子苷可降低肠黏膜屏障损害大鼠血清TNF-α、IL-1β、IL-6、D-LA、DAO及ET水平,增加IL-10水平（P<0.05,P<0.01）,减轻肠组织病理性损伤。结论 栀子苷可以减轻Zym导致的大鼠炎症反应,对Zym引起的肠黏膜屏障损害具有保护作用。     

阿尔泰瑞香提取物的体外抗炎及抗氧化活性

目的 探讨阿尔泰瑞香二氯甲烷提取物（Da-Dm）的体外抗炎及抗氧化活性。方法 以不同浓度（1、6、30 μg/mL）的Da-Dm作用于脂多糖（LPS,1 μg/mL）诱导的RAW264.7小鼠巨噬细胞,采用CCK-8法测定Da-Dm对细胞增殖的影响;Griess法检测细胞上清液中一氧化氮（NO）的释放量;ELISA法检测细胞上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）的分泌量。以维生素C为对照,采用FRAP法、DPPH法评价Da-Dm总抗氧化能力。结果 Da-Dm在浓度为30 μg/mL及以下时对RAW264.7细胞增殖没有影响。与LPS组比较,1~30 μg/mL的Da-Dm可明显降低LPS诱导的RAW264.7细胞分泌炎症因子NO、IL-1β、IL-6和TNF-α的含量（P<0.05,P<0.01）,并呈现浓度依赖性。Da-Dm对DPPH自由基有较好的清除能力,其清除率的IC50为318.1 μg/mL,在FRAP实验中其对Fe²⁺离子具有较强的还原能力。结论 Da-Dm可以抑制LPS诱导的RAW264.7细胞炎症反应,它的抗炎作用可能与抑制NO、IL-1β、IL-6和TNF-α的释放有关。Da-Dm具有较好的抗氧化活性。     

五倍子提取物对蛋鸡的产蛋性能和蛋品质及血液生化指标的影响

挑选1500只34周龄罗曼粉蛋鸡,随机均分为3个处理组,对照组饲喂基础饲粮,试验组分别饲喂添加了300 mg/kg的五倍子单宁酸(GTA)和没食子酸丙酯(PG)的基础饲粮,预试期1周,正试期8周,探究饲喂GTA和PG对产蛋高峰期蛋鸡的产蛋性能、蛋品质及血液生化指标的影响。结果表明：与对照相比,饲粮添加GTA和PG对蛋鸡产蛋性能无显著影响;饲粮添加GTA能显著改善试验8周时的蛋黄着色,但饲粮添加GTA和PG对蛋白高度、哈氏单位、蛋黄比例、蛋壳强度和蛋壳厚度均无显著影响;饲粮添加GTA能显著降低蛋鸡血浆中总蛋白(TP)、白蛋白(ALB)、球蛋白(GLB)和胆固醇(TC)的含量,有降低葡萄糖(GLU)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL–C)和低密度脂蛋白胆固醇(LDL–C)水平的趋势;饲粮添加PG能显著降低血浆TP和ALB水平,有降低GLU、GLB、TG、TC和HDL–C水平的趋势;饲粮添加GTA和PG对蛋鸡血清肝损伤指标(谷丙转氨酶、谷草转氨酶、碱性磷酸酶活性)和抗氧化指标(超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶活性和丙二醛含量)均无显著影响,但饲粮添加GTA能显著降低血清细胞炎症因子(白细胞介素–1、白细胞介素–6、肿瘤坏死因子–?)水平,饲粮添加PG可提高血清免疫球蛋白Y、M质量浓度。可见,在本研究条件下,饲粮中添加300 mg/kg的GTA和PG对罗曼粉蛋鸡的产蛋性能无显著影响,但GTA能提高蛋黄着色;饲粮添加GTA下调蛋鸡血脂、血浆蛋白及血清炎症因子水平的作用均大于PG的;饲粮添加PG可提升机体的免疫水平。     

A Sulfated Polysaccharide from Saccharina japonica Suppresses LPS-Induced Inflammation Both in a Macrophage Cell Model via Blocking MAPK/NF-κB Signal Pathways In Vitro and a Zebrafish Model of Embryos and Larvae In Vivo

Inflammation is a complicated host-protective response to stimuli and toxic conditions, and is considered as a double-edged sword. A sulfated Saccharina japonica polysaccharide (LJPS) with a sulfate content of 9.07% showed significant inhibitory effects against lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells and zebrafish. Its chemical and structural properties were investigated via HPLC, GC, FTIR, and NMR spectroscopy. In vitro experiments demonstrated that LJPS significantly inhibited the generation of nitric oxide (NO) and prostaglandin E₂ (PGE₂) via the downregulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression and suppressed pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-1β production via the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signal pathways in LPS-induced RAW 264.7 cells. Moreover, LJPS showed strong protective effects against LPS-induced inflammatory responses in zebrafish, increasing the survival rate, reducing the heart rate and yolk sac edema size, and inhibiting cell death and the production of intracellular reactive oxygen species (ROS) and NO. Its convenience for large-scale production and significant anti-inflammatory activity indicated the potential application of LJPS in functional foods, cosmetics, and pharmaceutical industries.     

The composition of the inflammatory infiltrate in three cases of polyneuritis equi

Polyneuritis equi (PNE) is a rare neurological disease in the horse. Because of the suspicion in PNE of a T-lymphocyte mediated immune response against the myelin, the objective of this study was to determine the composition of the inflammatory infiltrate in the involved nerves of 3 horses with PNE, studied retrospectively. T-lymphocytes were demonstrated in the lesions, which suggests a T-lymphocyte mediated immune response against myelin. In addition, the presence of B-lymphocytes indicated a local production of antibodies. More research, involving a prospective study, is needed to determine whether the T-lymphocytes are cytotoxic or T-helper lymphocytes.     

α-硫辛酸对热应激绵羊生长性能、抗氧化及免疫的影响

本试验旨在研究α-硫辛酸(α-LA)对热应激绵羊生长性能、抗氧化及免疫的影响,以期阐述其抗热应激的机理。选择4～5月龄、体重35～40 kg的绵羊15只,随机分为对照组(CTL)、低剂量组(LA-L,添加600 mg/kgα-LA)、高剂量组(LA-H,添加900 mg/kgα-LA),在热应激条件下(7～8月)饲喂40 d。结果表明:1)与对照组相比,α-LA添加组平均日采食量增加,低剂量组和高剂量组分别比对照组增加了7. 3%和2. 7%(P> 0. 05);高剂量组血清中丙氨酸转氨酶(ALT)、门冬氨酸转氨酶(AST)活性及α-LA添加组血浆中丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)含量显著下降(P<0.05);α-LA添加组血浆中三碘甲腺原氨酸(T3)、热休克蛋白70(HSP70)、免疫球蛋白G(IgG)含量及超氧化物歧化酶(SOD)活性显著升高(P <0.05)。2)与对照组相比,α-LA添加组肝脏中炎性相关因子IL-1β、IL-8、核因子-κB(NF-κB) mRNA表达量显著下降(P <0.05),抗炎因子干扰素-γ(IFN-γ)mRNA表达量显著升高(P<0.05)。综上,α-LA显著增强热应激绵羊的抗氧化能力,减少炎症相关因子的表达,增强抗炎因子的表达,提高机体免疫机能,且与α-LA剂量有关。     

Characteristic of Inflammatory Airway Disease in Japanese Thoroughbred Racehorses

Inflammatory airway disease (IAD) is a common cause of poor performance, interruption of training and premature retirement in racehorses. It is also reported that up to 80% of horses are affected at some point in the first years of training in UK and Australia. However, no studies with regard to the information on occurrence of IAD in Japanese Thoroughbred racehorses have been reported. To investigate the occurrence and the characteristics of IAD, epidemic research including endoscopic examination of the airway tract and trachea wash was conducted for Thoroughbred racehorses presenting coughs or poor performance which airway tract disease was suspected stalled in training facility managed by Japan Racing Association. Fifty-six out of 76 Thoroughbred racehorses (73.7%) presenting coughing or poor performance were diagnosed as IAD. Mean incidence rate of IAD was 0.3% and it has been confirmed that constant number of IAD exists in Japan. Up to 35.7% of IAD horses showed upper airway abnormalities in some extent. There was a trend for IAD horses to use wood shavings for bedding and fed hay from the ground compared with the control group. Therefore, improvement of stabling environment may aid in preventing IAD. This study demonstrated that Japanese Thoroughbred racehorses are affected by IAD likewise other countries as well as demonstrated the characteristics of IAD which may contribute to the clarification of the pathogenesis of IAD.