Effects of platelet-rich plasma on chondrocyte apoptosis and NLRP3/IL-1β signaling pathway in rabbits with osteoarthritis

AIM To explore the effect of platelet-rich plasma (PRP) on rabbit osteoarthritis and its possible mechanism. METHODS The rabbits with knee osteoarthritis were prepared and then divided into model group, sodium hyaluronate (SH) group and PRP group, and another sham operation group was set up, with 6 rabbits in each group. The gross morphological changes of rabbit cartilage were observed. HE staining was used to evaluate the pathomorphological changes of the cartilage. TUNEL staining was used to detect the apoptosis of chondrocytes. The expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/interleukin-1β （IL-1β） signaling pathway-related molecules was observed by immunohistochemical staining, and the protein levels of caspase-3, Bcl-2 and Bax were determined by Western blot. Chondrocytes were isolated and processed according to grouping, and the NLRP3 and IL-1β levels of the cells were measured by ELISA. RESULTS Compared with sham operation group, Pelletier score, Mankin score, chondrocyte apoptotic rate, the positive protein expression rates of NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax in model group were increased significantly (P<0.05), while the protein expression of Bcl-2 was decreased significantly (P<0.05). Compared with model group, Pelletier score, Mankin score, the apoptotic rate of chondrocytes, the positive protein expression rates of NLRP3, ASC, caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax in SH group and PRP group were decreased significantly (P<0.05), while the protein expression of Bcl-2 was increased significantly (P<0.05). In PRP group, Pelletier score, Mankin score, the apoptotic rate of chondrocytes, the positive protein expression rates of NLRP3, ASC, caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax were lower than those in SH group, while the protein expression of Bcl-2 was higher than that in SH group (P<0.05). Compared with control group, the expression of NL?RP3 and IL-1β in MCC950 (NLRP3 ihibitor) group were significantly reduced (P<0.05), the expression of NLRP3 in eucalyptol (IL-1β inhibitor) group was not significantly changed (P>0.05), and the expression of IL-1β was significantly reduced (P<0.05). CONCLUSION Platelet-rich plasma promotes the repair of cartilage in osteoarthritis rabbits, which has better effect than SH. The mechanism may be related to the inhibition of NLRP3/IL-1β pathway and the reduction of chondrocyte apoptosis.     

黑种草子提取物对小鼠酒精性肝损伤的修复作用及机制研究

【目的】验证黑种草子提取物对小鼠酒精性肝损伤的修复作用并探究其机制,为其临床开发研究提供理论依据。【方法】将56只3周龄昆明小鼠随机均分为7组：空白对照组,模型组,黑种草子提取物高、中、低剂量组,黑种草子粉剂组及水飞蓟宾阳性对照组,每组8只。除空白对照组外,其余各组均以10 mL/kg 60%酒精灌胃,每天1次,连续15 d。第16天,黑种草子提取物高、中、低剂量组分别给予8、4、2 mL/kg黑种草子提取物,每天1次,连续10 d后处死小鼠。计算各组小鼠肝脏指数;检测血清中谷丙转氨酶（alanine aminotransferase,ALT）、谷草转氨酶（aspartate aminotransferase,AST）、过氧化氢酶（catalase,CAT）、超氧化物歧化酶（superoxide dismutase,SOD）和谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-Px）活力;制备肝脏石蜡切片观察其病理变化;Western blotting检测NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3,NLRP3）、含半胱氨酸的天冬氨酸蛋白水解酶1（cysteinyl aspartate specific proteinase 1,Caspase-1）和白细胞介素1β（interleukin-1β,IL-1β）的蛋白表达水平。【结果】与空白对照组相比,模型组小鼠肝脏指数显著上升（P<0.05）,肝细胞出现明显颗粒变性,水泡变性,核溶解,界限区分不清,肝索消失;血清ALT、AST活力显著上升（P<0.05）,SOD、CAT、GSH-Px活力显著下降（P<0.05）,说明肝损伤模型建立成功,且肝脏组织中NLRP3、Caspase-1和IL-1β蛋白表达水平显著升高（P<0.05）。与模型组相比,黑种草子提取物各剂量组血清ALT、AST活力均呈下降趋势,SOD、CAT、GSH-Px活力均呈上升趋势,其中高剂量组上述各指标均呈显著差异（P<0.05）,肝脏组织结构明显改善,NLRP3、Caspase-1和IL-1β蛋白表达水平均显著下降（P<0.05）。【结论】高剂量（8 mL/kg）黑种草子提取物可显著提高肝损伤小鼠的抗氧化水平,且可以通过抑制NLRP3炎性小体的表达而减少IL-1β的合成,从而改善肝脏功能和修复受损的肝脏组织结构。     

猪ESR、FSH-β和PRLR基因合并基因型对产仔数的影响

摘 要: 本试验以辽宁省三个引进猪种：长白、大白和杜洛克为对象,采用PCR-RFLP方法检测ESR、FSH-β和PRLR基因的多态性,分析三个基因的单基因及合并基因型对产仔数的影响。结果表明,ESR 、FSH-β和PRLR基因都存在AA、AB和BB三种基因型,在各猪群内基因型分布差异不显著(P＞0.05),各基因均处于遗传平衡状态。单基因分析表明,ESR 和FSH-β基因型产仔数的最小二乘均值具有按AA、AB 、BB递增的趋势,BB型为优良基因型,而PRLR基因的优良基因型为AA型。ESR 、FSH-β和PRLR基因的合并分析表明BBBBAA为最优合并基因型,合并后效应显著（P＜0.05)或极显著（P＜0.01),且合并基因型的遗传效应高于单基因型效应。因此,可以利用合并基因型对猪的产仔数进行标记辅助选择。     

β-甘露聚糖酶对AA+鸡生产性能和免疫功能的影响

为了探讨日粮中添加β-甘露聚糖酶对AA+鸡生产性能和免疫功能的影响,选取1日龄健康的AA+肉鸡180羽,随机为2个处理组,每个处理3个重复,每个重复30只。试验分为对照组(基础日粮)和试验组（基础日粮+ 400g/Tβ-甘露聚糖酶）,试验期42天。结果表明β-甘露聚糖酶能不同程度的提高AA鸡各生长阶段的日增重和饲料转化率,但差异显著（P>0.05）,明显提高42d血液中免疫球蛋白IgM、IgG、IgA的含量,差异显著。21d、42d的免疫器官指数、T淋巴细胞转化率和ND抗体水平也有不同程度的提高,与对照组相比,差异不显著（P>0.05）。说明β-甘露聚糖酶能提高肉用鹌鹑生长性能和增强免疫的功能。     

野生与家养乌苏里貉FSHβ和FSHR基因的克隆及序列分析

对野生和家养乌苏里貉的FSHβ基因部分序列、FSHR基因5'端上游调控区和外显子10进行扩增和测序,获得序列长度分别为1257、757和1398 bp,提交GenBank,登录号分别为HQ385977、HQ385978和HQ385979。BLAST比较分析3段序列外显子区,与犬的同源性分别为99%、98%和98%。利用DNAMAN软件对2只野生与2只家养貉进行多序列比对,结果分析,3个基因片段中分别检测到12、10和3个变异位点,其中编码区碱基的突变位点分别有2、1和0个;将2只野生貉的序列相比较,发现3段序列分别有6、9和3个多态位点,而将2只家养貉的序列相比后分别有2、1和0个多态位点,说明野生貉更具有遗传多样性。     

中药HDβ-环糊精包合物的制备及性质研究

目的：研究中药HDβ-环糊精（β-CD）包合物的制备工艺及其药效研究。方法：选用饱和溶液法制备中药HD的包合物,采用正交试验法对包合物的制备条件进行优化。结果：制备中药HDβ-CD包合物的最佳工艺条件为中药HDβ-CD投料质量比1∶3,反应温度50℃,搅拌时间4h,搅拌速度1500 r/min,包合率为35.56%。结论：该法包合效果较好,掩盖了药物的苦味,改善了药物的稳定性,且其药效没有发生改变。     

网渔具计算机数值模拟的研究进展

对近年来网渔具的计算机数值模拟中使用的几何拓扑分解方法、数学建模思想和模型算法等进行了归纳和系统整理,并探讨了各种算法的稳定性和收敛性。分析认为:刚体有限元法数学建模的优点是模型简化,能基本反映网具粗糙形状,缺点是忽略网线张力,无法精确反映网具的受力和网具的运动状态;基于质量弹簧体系和牛顿力学的柔性有限元法是主流,数值模拟结果更符合网具的实际状况,网形更细腻逼真,计算结果与网具实测拟合较好;显式算法对时间步长较敏感,稳定性和精度不如隐式算法好。NEWTON-RAPHSON和NEWMARK-β算法具有无条件稳定性和收敛性,大部分研究采用了此算法。     

Metabonomic study of the biochemical profiles of heterozygous myostatin knockout swine

Myostatin is a transforming growth factor-βfamily member that normally acts to limit skeletal muscle growth. Myostatin gene(MSTN) knockout(KO) mice show possible effects for the prevention or treatment of metabolic disorders such as obesity and type 2 diabetes.We applied chromatography and mass spectrometry based metabonomics to assess system-wide metabolic response of heterozygous MSTN KO(MSTN+/–) swine. Most of the metabolic data for MSTN+/–swine were similar to the data for wild type(WT) control swine. There were, however,metabolic changes related to fatty acid metabolism,glucose utilization, lipid metabolism, as well as BCAA catabolism caused by monoallelic MSTN depletion.The statistical analyses suggested that:(1) most metabolic changes were not significant in MSTN+/–swine compared to WT swine;(2) only a few metabolic properties were significantly different between KO and WT swine,especially for lipid metabolism. Significantly, these minor changes were most evident in female KO swine and suggested differences in gender sensitivity to myostatin.     

黏虫体内两种微管蛋白基因cDNA序列的克隆与序列分析

以黏虫4龄幼虫为材料提取总RNA,利用RT-PCR和cDNA末端快速扩增技术(RACE),分别扩增得到该虫的α和β微管蛋白基因的cDNA序列各1条。其中α微管蛋白基因的cDNA序列1 443个碱基,包括一个1 353个碱基的开放阅读框,编码一个含450个氨基酸的蛋白,分子量约为50.0ku。氨基酸的142~148位存在一个微管蛋白标志信号片段GGGTGSG,在氨基酸序列的C-端有一个酪氨酸残基,N-端存在一个对转录后调控非常重要的保守区MRECI序列,以上特点与其他昆虫α微管蛋白氨基酸序列相同。黏虫β微管蛋白基因cDNA序列含1 906个碱基,开放读码框1 344个碱基,编码氨基酸447个,分子量约为50.2ku,等电点4.75。1~4个氨基酸MREI为β微管蛋白转录后调控信号,140~146GGGTGSG位同样存在一个微管蛋白标志信号片段。序列比对表明,克隆的α和β微管蛋白基因与其他昆虫的α和β微管蛋白基因在核苷酸和氨基酸水平上都是高度同源的,黏虫与家蚕(Bombyx mori)α微管蛋白的氨基酸序列同源性达到99.3%,与其他3种夜蛾科昆虫α微管蛋白的氨基酸序列同源性更是达到100%。黏虫与家蚕β微管蛋白的氨基酸序列同源性达到98.7%,与烟草天蛾β微管蛋白的氨基酸序列同源性达到99.6%。两个基因的cDNA序列已经登录GenBank并获得登录号分别为EU100016和EU234504。     

文昌鸡促卵泡激素β基因内含子1的SNP分析及其与早期产蛋性能的相关性研究

以109只海南文昌鸡为试验动物,利用PCR-SSCP技术检测了促卵泡激素(follicle-stimulating hormone β,FSHβ)基因的多态性,在其内含子1区域内发现了1个SNP,在群体中检测出2种基因型:AA、AB。将不同基因型个体与产蛋数、畸形蛋数、体重和开产日龄等进行了关联分析。结果表明,AA基因型鸡个体的产蛋数比AB基因型个体多9.54枚,畸形蛋数比AB基因型个体少1.54枚,开产日龄比AB基因型个体早2.97 d,且差异均不显著(P＞0.05)。22~26、27~30和31~34周龄的AA基因型鸡的产蛋量显著高于AB基因型(P＜0.05),而18~21和35~39周龄的AA基因型鸡的产蛋量也明显高于AB基因型,但差异不显著(P＞0.05)。AA基因型鸡6、8、14、20周龄的体重高于AB基因型个体,AA基因型个体18周龄的体重却低于AB基因型个体,差异均不显著(P＞0.05)。总体而言,FSHβ基因位点的A等位基因对产蛋数、畸形蛋数、体重和开产日龄的影响均未达到差异显著水平。