Pyroptosis mediates high glucose-induced inflammation and injury in MC3T3-E1 osteoblasts

AIM To investigate whether pyroptosis contributes to the inflammation and injury in mouse embryonic osteoblastic cell line MC3T3-E1 induced by high glucose (HG; 45 mmol/L glucose). METHODS The cell viability was measured by CCK-8 assay. The protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and caspase-1 (CASP1) were determined by Western blot. The secretion levels of interleukin-18 (IL-18) and IL-1β were measured by ELISA. The intracellular level of reactive oxygen species (ROS) was detected by 2',7'-dichlorodihydrofluorescein diacetate staining followed by photofluorography. Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorography. The alkaline phosphatase (ALP) activity was determined using the ALP kit, and the number of mineralized nodules was detected by alizarin red S staining. RESULTS After the MC3T3-E1 osteoblasts were treated with HG for 24 h, the protein expression levels of NLRP3 and CASP1, and the secretion levels of IL-18 and IL-1β were significantly increased. The decrease in cell viability, and the increases in ROS generation and MMP loss were also observed. Moreover, the differentiation and mineralization of MC3T3-E1 osteoblasts were inhibited, evidenced by decreases in both ALP activity and mineralized nodule number. Knockdown of CASP1 by siRNA attenuated the HG-induced osteoblast inflammation and injury mentioned above. CONCLUSION Pyroptosis mediates HG-induced inflammation and injury in MC3T3-E1 osteoblasts.     

Mouse melanoma cell exosomes promote expression of Rac1 protein in fibroblasts

AIM To investigate the effect of exosomes secreted by mouse melanoma cells on the expression of Ras-related C3 botulinum toxin substrate 1 (Rac1) protein in fibroblasts. METHODS Ultracentrifugation was adopted to separete exosomes secreted by mouse melanoma B16-F10 cells. The morphological structure of exosomes was observed by negative-staining electron microscopy. The size distribution of exosomes was determined by nanoparticle tracking analysis (NTA). The exosomal markers, tumor susceptibility gene 101 (Tsg101) and tyrosinase-related protein 2 (Tyrp2), were identified by Western blot. Laser confocal microscopy was used to observe the process that mouse embryonic fibroblasts (MEF) took in exosomes during co-culture. Immunocytochemical staining and Western blot were used to detect the expression of Rac1 protein in MEF. RESULTS B16-F10 cell exosomes showed a typical tea tray-like structure, with a size range of 141~255 nm, and expressed protein markers Tsg101 and Tyrp2. The results of laser confocal microscopy showed that compared with co-culture at 0 h, a small number of exosomes appeared in the MEF at 12 h, and a large number of exosomes accumulated in the MEF after co-cultured for 24 and 36 h. Western blot analysis showed that compared with co-culture at 0 h, the expression of Rac1 protein in the MEF was significantly increased at 24 h and 36 h of co-culture (P<0.01). The results of immunocytochemical staining showed that compared with co-culture at 0 h, the positive expression level of Rac1 in the MEF cells was significantly increased at 12 h, 24 h and 36 h of co-culture (P<0.05 or P<0.01). CONCLUSION Intake of exosomes secreted by mouse melanoma cells promotes the expression of Rac1 protein in fibroblasts.     

番茄匍柄霉叶斑病拮抗细菌的筛选与鉴定

应用平板对峙法从连作多年的番茄根际土样中筛选得到对番茄匍柄霉叶斑病具有较强拮抗活性的细菌菌株ZF161，该菌株对番茄匍柄霉的平板抑制率达到70.51%。离体叶片试验显示，菌株ZF161对番茄匍柄霉叶斑病防治效果达到69.83%。通过菌落形态观察、生理生化特性、Biolog测定和多基因系统发育树综合分析，鉴定菌株ZF161为枯草芽孢杆菌（Bacillus subtilis）。番茄盆栽试验结果表明，菌株ZF161对番茄匍柄霉叶斑病的防治效果达到63.27%。进一步平板对峙抑菌谱试验结果显示，菌株ZF161对其他7种病原真菌也具有较好的抑制效果。上述结果说明，菌株ZF161对番茄匍柄霉叶斑病具有良好的防治效果，且对多种病原真菌也具有抑制作用，具有开发应用的潜力。     

Soluble Klotho protein suppresses THP-1-derived foam cell formation

AIMTo investigate the role of soluble Klotho protein in THP-1-derived foam cell formation. METHODSTHP-1 monocytes were induced into macrophages by treatment with 160 nmol/L phorbol myristate acetate for 48 h, and then were divided into 6 groups: negative control group (THP-1-derived macrophages), positive control group [THP-1-derived foam cells induced by oxidized low-density lipoprotein (ox-LDL) for 48 h], and 25, 50, 100 and 200 μg/L soluble Klotho protein groups (THP-1-derived macrophages pretreated with soluble Klotho protein at the indicat?ed concentraions for 2 h and then induced by ox-LDL for 48 h). Lipid droplets in cytoplasm were observed by oil red O staining. The cholesterol outflow rate was detected by scintillation counting technique. The content of intracellular total cholesterol, free cholesterol and cholesterol ester was detected by enzyme fluorescence analysis. The expression of acyl-coenzyme A:cholesterol acyltransferase 1 （ACAT1） and ATP-binding cassette transport?er A1 (ABCA1) at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. RESULTSOil red O staining and lipid mass quantification showed that THP-1-derived foam cell formation was dose-dependently suppressed by soluble Klotho protein. The cholesterol efflux rate of THP-1-derived foam cells was increased by soluble Klotho protein in a dose-dependent manner (P<0.05). In addition, soluble Klotho protein decreased the expression of ACAT1 and increased the expression of ABCA1 in a dose-dependent manner (P<0.05). CONCLUSION The soluble Klotho protein inhibits THP-1-derived foam cell formation in a dose-dependent manner by down-regulating the expression of ACAT1 and up-regulating the expression of ABCA1.     

栅藻B38营养成分分析及蛋白质营养价值评价

对栅藻B38的营养成分进行了全面分析,结果表明,栅藻藻粉中油脂含量14%,总糖含量13.86%,粗纤维3.52%,灰分4.28%,蛋白质49.0%,为高蛋白质类微藻。栅藻B38中含有23种脂肪酸。栅藻B38中氨基酸总量为852.58 mg/g,必需氨基酸占总氨基酸的47.45%,必需氨基酸指数为85.42%。两种模式的氨基酸比值系数分较接近,均在65以上;待评价物质的氨基酸品质与模式蛋白的贴近度接近1(0.93和0.88)。栅藻B38中共检测出23种矿物质元素,其中钠、钾、钙、镁、铁、铝含量较为丰富。重金属的含量均低于国家食品卫生标准。栅藻B38中维生素B3(106.02 mg/100 g)和维生素E(540.77 mg/100 g)的量较高。综上,栅藻B38的营养较丰富,可以进一步开发利用。     

Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.     

四川地区猕猴桃病毒1的RT-PCR检测及外壳蛋白基因序列分析

为了明确近期新西兰报道的侵染猕猴桃的新病毒——猕猴桃病毒1（Actinidia virus 1，AcV-1）在四川地区猕猴桃上的发生情况及其分子特性，采用RT-PCR对来自四川6个地区疑似感染病毒的90份猕猴桃主栽品种‘红阳’和‘金果’的叶片样品进行检测。结果表明，22份样品为AcV-1阳性，检出率为24.4%。将获得的5个AcV-1分离物和已报道的新西兰分离物K75的外壳蛋白（coat protein，CP）基因序列进行比对，结果表明，分离物间核苷酸序列和氨基酸序列的相似性分别为84.8% ~ 97.1%和89.7% ~ 99.6%，其中除邛崃分离物HYH5与新西兰分离物K75的核苷酸序列相似性较高（97.1%）外，其余4个分离物均低于91%。系统进化分析结果显示，这些分离物主要聚集在3个分支上，分离物HYH5与新西兰分离物K75位于同一分支，其余分离物位于另外2个分支。     

不同光质照射下真姬菇的生长发育与光受体的响应表达

采用 5 种不同光质光源，分别对真姬菇（Hypsizygus marmoreus）的菌丝、原基和子实体进行照射，检测菌丝生长速度、原基形成数量、子实体菌柄长度、菌柄直径、菌盖直径以及产量等。结果显示：真姬菇菌丝在黑暗和红光下生长较快，在红绿蓝复合光下生长最慢；在蓝光下形成的原基数量最多，而在红光下原基数量最少；子实体阶段，黑暗下菌柄的长度及直径最大，绿光下菌柄长度及直径最小；而在蓝光和红绿蓝复合光下菌盖直径最大。相同培养时间，真姬菇在黑暗和蓝光下产量较高，在绿光下产量最低。进一步检测真姬菇光受体基因 WC1 和 WC2 在不同光质条件下的差异表达。结果表明，在菌柄中 WC1 在黑暗下表达量最高，WC2 在蓝光下表达量最高；而在菌盖中 WC1 和 WC2 均在蓝光下表达量最高。     

LED补光组合对大棚越橘生长发育的影响

以2年生南高丛越橘‘Emerald’为材料，以大棚内自然光作为对照，研究LED光源的红蓝光（3︰1和6︰1）、紫外光（UVA）对植株长势、叶片光合作用、碳氮代谢、开花基因表达及开花率、果实品质的影响。结果表明：红蓝光组合处理下‘Emerald’的植株营养生长较旺盛，株高、1年生枝条长度和粗度显著高于对照。此外，红蓝光（6︰1）处理下叶片的叶绿素相对含量、比叶重和净光合速率均显著提高。红蓝光组合也可诱导植株开花，开花基因FT表达量和开花率明显高于对照。紫外光下叶片的氮含量、开花率及FT基因表达量显著高于对照。不同光质组合补光对‘Emerald’的果实品质有显著影响，红蓝光（3︰1）处理下果实的质量、横纵径、可溶性固形物、可溶性糖、花青素含量和糖酸比均高于对照。总之，不同光质补光会促进越橘‘Emerald’的生长发育，红蓝光6︰1组合对促进营养生长作用相对较大，红蓝光3︰1组合对提高果实品质效果较好。紫外光虽能改变植株形态和促进开花，但果实品质提高效果较红蓝光处理稍弱。     

生物饵料对赤点石斑鱼不同养殖阶段的影响

通过对赤点石斑鱼(Epinephelus akaara)亲本强化、苗种繁育阶段的饵料进行研究,结果表明:采用牡蛎作为赤点石斑鱼亲本营养强化饵料使得亲本成活率达89.33%,所产的卵子平均受精率达78.9%、孵化率达80.1%,均显著高于使用南美白对虾和蛏子组(P<0.05);苗种培育至摄氏卤虫阶段,采用鱼油(4 g/L)与经小球藻强化轮虫和虾青素(5 g/L)混合代替,成活率29.53%,全长增长率175.00%,效果最佳;苗种培育至摄氏桡足类阶段,采用鱼油(4 g/L)与桡足类和虾青素(5 g/L)培育,成活率和全长增长率均显著高于其他组(P<0.05);而后期采用软颗粒料(鳗粉:鳀鱼=3:2,添加虾青素5 g/kg)可极显著提高赤点石斑鱼的养成成活率和全长增长率(P<0.01)。