Protein kinase C delta null mice exhibit structural alterations in articular surface,intra-articular and subchondral compartments

IntroductionStructural alterations in intra-articular and subchondral compartments are hallmarks of osteoarthritis, a degenerative disease that causes pain and disability in the aging population. Protein kinase C delta (PKC-δ) plays versatile functions in cell growth and differentiation, but its role in the articular cartilage and subchondral bone is not known.MethodsHistological analysis including alcian blue, safranin O staining and fluorochrome labeling were used to reveal structural alterations at the articular cartilage surface and bone–cartilage interface in PKC-δ knockout (KO) mice. The morphology and organization of chondrocytes were studied using confocal microscopy. Glycosaminoglycan content was studied by micromass culture of chondrocytes of PKC-δ KO mice.ResultsWe uncovered atypical structural demarcation between articular cartilage and subchondral bone of PKC-δ KO mice. Histology analyses revealed a thickening of the articular cartilage and calcified bone–cartilage interface, and decreased safranin O staining accompanied by an increase in the number of hypertrophic chondrocytes in the articular cartilage of PKC-δ KO mice. Interestingly, loss of demarcation between articular cartilage and bone was concomitant with irregular chondrocyte morphology and arrangement. Consistently, in vivo calcein labeling assay showed an increased intensity of calcein labeling in the interface of the growth plate and metaphysis in PKC-δ KO mice. Furthermore, in vitro culture of chondrocyte micromass showed a decreased alcian blue staining of chondrocyte micromass in the PKC-δ KO mice, indicative of a reduced level of glycosaminoglycan production.ConclusionsOur data imply a role for PKC-δ in the osteochondral plasticity of the interface between articular cartilage and the osteochondral junction.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-015-0720-4) contains supplementary material, which is available to authorized users.     

Enhancement of Astragalus polysaccharide on the immune responses in pigs inoculated with foot-and-mouth disease virus vaccine

The effects of Astragalus polysaccharides (APS) on the immune response in pigs immunized with foot-and-mouth disease virus (FMDV) vaccine were investigated. Fifteen pigs were randomly divided into five groups. Four groups were vaccinated with a FMDV inactivated vaccine. Pigs in three experimental groups were administered varying doses of APS (APS1, 5 mg/kg; APS2, 10 mg/kg; APS3, 20 mg/kg). The influence of APS on the number of CD3⁺CD4⁻CD8⁺ cytotoxic T cells, CD3⁺CD4⁺CD8⁺ T helper memory cells, and CD3⁻CD4⁻CD8⁺ natural killer cells among peripheral blood lymphocytes (PBL) in the three APS groups were significant compared to the vaccine group. In vitro stimulation of PBL by Con A and LPS in APS groups induced a stronger proliferative response at 2 and 6 weeks post-inoculation (PI). APS markedly increased the titer of FMDV-specific antibody in a dose-dependent manner, and up-regulated mRNA expression of IFN-γ and IL-6. APS could potentially be used as an immunomodulator for a FMDV vaccine and provide better protection against FMDV.     

Functional comparison of metallothioneins MTT1 and MTT2 from Tetrahymena thermophila

Metallothioneins MTT1 and MTT2 from Tetrahymena thermophila have been characterized. The MTT1 contains mainly characteristic Cys-Cys-Cys and Cys-Cys clusters, but MTT2 contains mainly Cys-X-Cys cluster. Cd₁₆-MTT1 mainly consists of α-helix and β-turns, in contrast, Cd₁₁-MTT2 mainly consists of random coils. Reaction of Cd₁₆-MTT1 and Cd₁₁-MTT2 with nitric oxide leads to intramolecular disulfide bond formation, respectively. Binding stabilities of Cd²⁺, Hg²⁺ and Zn²⁺ to MTT1 are stronger than those to MTT2. Cu²⁺ can not replace Cd²⁺ from Cd₁₆-MTT1 complex, but can replace Cd²⁺ from Cd₁₁-MTT2 complex. The analysis of qRT-PCR revealed MTT2 mRNA levels were 31-fold higher than those of MTT1 under basal conditions. These results further suggest MTT1 possibly play a role in the detoxification of heavy metal ions, and MTT2 may be involved in the homeostasis of copper ions.     

Effects of dissolved organic matter from sewage sludge on the atrazine sorption by soils

The effects of dissolved organic matter (DOM), water soluble organic matter derived from sewage sludge, on the sorption of atrazine (2-chloro-4-ethylamino-6-isopropylamino-1,3,5-trazine) by soils were studied using a batch equilibrium technique. Six paddy soils, chosen so as to have different organic carbon contents, were experimented in this investigation. Atrazine sorption isotherms on soils were described by the linear equation, and the distribution coefficients without DOM (Kd) or with DOM (Kd*) were obtained. Generally, the values of Kd*/Kd initially insoil-solution system form. Critical concentrations of DOM (DOMnp) were obtained where the value of Kd* was equal to Kd. The presence of DOM with concentrations lower than DOMnp promoted atrazine sorption on soils (Kd* ＞ Kd), whereas the presence of DOM with concentrations higher than DOMnp tended to inhibit atrazine sorption (Kd* ＜ Kd). Interestingly, DOMnp for tested soils was negatively correlated to the soil organic carbon content, and the maximum of Kd*/Kd (i.e.Kmax) correlated positively with the maximum of DOM sorption on soil (Xmax). Further investigations showed that the presence of hydrophobic fraction of DOM evidently promoted the atrazine sorption on soils, whereas the presence of hydrophilic DOM fraction obviously tended to inhibit the atrazine sorption. Interactions of soil surfaces with DOM and its fractions were suggested to be the major processes determining atrazine sorption on soils. The results of this work provide a reference to the agricultural use of organic amendment such as sewage sludge for improving the availability of atrazine in soils.     

效应蛋白LepB的表达,纯化及其亚克隆片段的结晶研究

为了研究嗜肺军团菌通过其效应蛋白发挥的致病机理,效应蛋白的结构和生物学功能研究是至关重要的。我们构建了效应蛋白LepB全长载体PFastBac1,应用Bac-to-Bac杆状病毒表达系统成功表达了全长LepB蛋白,并构建9个LepB亚克隆的原核表达载体PGEX-1,通过GST亲和层析,离子交换层析,凝胶过滤层析,纯化得到了纯度较高的蛋白片段。通过悬滴气相扩散法进行蛋白结晶筛选,获得了效应蛋白LepB片段480~679的蛋白晶体,为解析效应蛋白LepB的结构和生物学功能研究奠定基础。     

Generation and Analysis of ESTs from the Grass Carp,Ctenopharyngodon idellus

Grass carp, Ctenopharyngodon idellus (Valenciennes, 1844), is an economically important species widely cultured in the world, but its genome research resources are largely lacking. The objectives of this study were to construct normalized cDNA libraries for efficient EST analysis, to generate ESTs from these libraries, and to identify EST-related molecular markers such as microsatellites and single nucleotide polymorphisms (SNPs) for genetic analysis of this species. A total of 6,269 ESTs were generated representing 4,815 unique sequences, from which 105 putative microsatellites and 5,228 SNPs were identified. These genome resources provide the material basis for future genetic and functional analyses in this species.     

紫茎泽兰的化学成分初报

紫茎泽兰(Eupalorium adenophorum Spreng)原产中美墨西哥,现在滇南一带广泛分布,对林、牧业生产造成严重危害。其化学成分研究未见报道。 从紫茎泽兰的叶和花序中,分到九个单体,经详细的光谱解析和与标准品对照,其中五个成分的化学结构分别为:正三十二烷n-dotriacontane(1),β-谷甾醇β-sitosterol(2),豆甾醇stigmasterol(3),蒲公英醇棕榈酸酯taraxasteryl palmitate(4),蒲公英醇乙酸酯taraxastcryl acetate(5)。     

嘌呤拟似物对豚鼠胃窦环行肌自发性收缩及电活动的影响

为探讨非肾上腺素能非胆碱能神经递质对胃窦环行肌功能的调节作用,在离体胃平滑肌上观察了嘌呤拟似物对胃窦环行肌自发性收缩活动和电活动的影响。电活动用传统的细胞内微电极记录,并和收缩活动同步描记于多道生理记录仪。结果表明,嘌呤能P2Y受体激动剂,三磷酸腺苷(ATP)和2-methylthio ATP(2-MeSATP)均增强胃窦平滑肌的收缩活动,但不影响电活动,而且ATP和2-MeSATP对胃平滑肌收缩活动的增强作用可被嘌呤能P2Y受体阻断剂,reactive blue-2和苏拉明(suramin)所阻断。用100μmol／L α,β-MeATP引起的脱敏感使P2X受体被阻断,ATP增强胃窦平滑肌收缩活动的效应不受影响。嘌呤能P2X受体激动剂,α,β-MeATP明显抑制胃窦环行肌自发性收缩活动,同时使膜电位明显超极化。ATP对胃窦平滑肌的收缩作用不被L型钙通道阻断剂尼卡地平(nicardipine)阻断,但细胞外用无钙液灌流时这种效应则完全被阻断。用前列腺素合成抑制剂消炎痛预先处理20min后,ATP和2-MeSATP仍能增强胃窦平滑肌的自发性收缩活动。以上结果提示：(1)ATP和2-MeSATP通过嘌呤能P2Y受体增强胃窦平滑肌的自发性收缩活动,而α,β-MeATP或β,γ-MeATP通过嘌呤能P2X受体使膜电位超极化,引起胃窦平滑肌的舒张;(2)ATP和2-MeSATP增强胃窦平滑肌自发性收缩活动的效应依赖于细胞外钙,但钙离子进入细胞的途径并不是电压依赖性钙通道;(3)ATP和2-MeSATP增强胃窦平滑肌自发性收缩活动的效应不通过前列腺素介导。