Identifying and Characterizing a Novel Peritrophic Matrix Protein (MdPM-17) Associated With Antibacterial Response From the Housefly,Musca domestica (Diptera: Muscidae)

Peritrophic matrix/membrane (PM) critically prevents the midgut of insects from external invasion by microbes. The proteins in the peritrophic membrane are its major structural components. Additionally, they determine the formation and function of this membrane. However, the role of PM proteins in immune regulation is unclear. Herein, we isolated a novel PM protein (MdPM-17) from Musca domestica larvae. Further, the function of MdPM-17 in regulating host innate immunity was identified. Results showed that the cDNA of MdPM-17 full is 635 bp in length. Moreover, it consists of a 477-bp open reading frame encoding 158 amino acid residues. These amino acid residues are composed of two Chitin-binding type-2 domain (ChtBD2) and 19 amino acids as a signal peptide. Moreover, tissue distribution analysis indicates that MdPM-17 was enriched expressed in midgut, and moderate levels in the fat body, foregut, and malpighian tubule. Notably, MdPM-17 recombinant protein showed high chitin-binding capacity, thus belongs to the Class III PM protein group. MdPM-17 protein silencing via RNA interference resulted in the expression of antimicrobial peptide (defensin, cecropins, and diptericin) genes, and this occurred after oral inoculation with exogenous microbes Escherichia coli (Enterobacteriales:Enterobacteriaceae), Staphylococcus aureus (Bacillales:Staphylococcaceae), and Candida albicans (Endomycetales:Saccharomycetaceae)). Therefore, all the antimicrobial peptide (AMP) gene expression levels are high in MdPM-17-depleted larvae during microbial infection compared to controls. Consequently, these findings indicate that MdPM-17 protein is associated with the antibacterial response from the housefly.     

不同光源对4种蔬菜幼苗叶绿素荧光及生理指标的影响

为了解不同光源或光源组合对茄子、辣椒、番茄、黄瓜幼苗叶绿素荧光及生理指标的影响,在4种基本光源的基础上,比较了不同光源处理下不同蔬菜植株叶片的叶绿素荧光参数、可溶性蛋白和可溶性糖含量。结果表明:红蓝光源不利于4种蔬菜幼苗叶片光合色素的形成,茄子、辣椒、番茄和黄瓜叶片中的光合色素含量分别在2白4红2黄、3白3红2黄、8黄或8红、8白光源下最高;茄子和番茄幼苗叶片在红蓝光源下,辣椒和黄瓜在3白3红2黄光源下,最大光化学效率和实际光化学效率都最大;茄子和辣椒幼苗在红蓝光源下所受非生物胁迫最小,叶片中可溶性蛋白含量最低,而番茄和黄瓜分别在3白3红2黄和8白光源下的可溶性蛋白含量最低;红蓝光源可明显增加4种蔬菜幼苗叶片的可溶性糖含量,对幼苗的形态建成有积极的影响。在冬春季蔬菜育苗生产中,可根据不同光源优势为不同种类蔬菜组配光源,以促进幼苗生长发育。     

利用荧光观测明确载球孢白僵菌松毛虫赤眼蜂对亚洲玉米螟的防控增效作用

载球孢白僵菌松毛虫赤眼蜂能够显著提高对亚洲玉米螟的防控效果,为了明确其对亚洲玉米螟卵期及幼虫期可持续防控的作用机理,采用松毛虫赤眼蜂吸附绿色荧光蛋白（GFP）标记的球孢白僵菌分生孢子,进行赤眼蜂载菌情况,以及亚洲玉米螟卵和幼虫的寄生、侵染过程观察。结果表明,赤眼蜂羽化后能够吸附柞蚕卵表面粘附的白僵菌分生孢子,并将其携带至亚洲玉米螟卵块表面,并吸附于未被赤眼蜂寄生的亚洲玉米螟卵孵化的幼虫体表,实现侵染并致死,幼虫带菌率达60.00%,网室内杀虫生物测定僵虫率达27.00%。本研究表明,载菌赤眼蜂在提高杀虫效率的同时实现害虫可持续防控,该方法为其他载菌天敌的应用提供了依据。     

生物炭对Bt Cry1Ac蛋白抗紫外能力影响

为了研究生物炭对紫外线的防护作用,以豆壳烧制的生物炭作为载体,研究生物炭对Bt Cry1Ac蛋白的吸附行为以及生物炭对Cry1Ac蛋白的紫外保护作用。使用扫描电子显微镜、透射电子显微镜、X-射线粉末衍射以及傅立叶红外光谱等手段对生物炭的形貌和结构进行表征。结果表明,生物炭是典型的多孔结构材料,表面具有丰富的官能团。Cry1Ac蛋白与生物炭吸附平衡时间为50 min,最合适的吸附浓度比（生物炭：蛋白）为1：100,二者吸附符合准二级动力学模型和Langmuir模型。在UVB紫外照射4 h后,生物炭与Cry1Ac蛋白复合物对棉铃虫的生物活性是单纯蛋白的4.93倍,显示生物炭具有较好的紫外抵抗效果。研究结果初步表明,制备得到的生物炭能够显著提高Cry1Ac蛋白的抗紫外能力,为后续研发耐受紫外线的农药剂型提供新材料。     

Effects of calpain-2 and autophagy-related protein 5 on hepatocyte apoptosis induced by endoplasmic reticulum stress

AIMTo investigate the effects of calpain-2 and autophagy-related protein 5 (Atg5) on apoptosis of BRL-3A rat normal liver cells during endoplasmic reticulum stress (ERS) induced by dithiothreitol (DTT). METH?ODS: BRL-3A cells were treated with DTT at 2.0 mmol/L for 0, 6, 12 and 24 h to induce ERS. Real-time cell analysis (RTCA) was used to measure the effect of DTT on BRL-3A cell proliferation. Apoptosis and cell cycle distribution were analyzed by flow cytometry. The mRNA expression of calpain-2 and Atg5 was detected by real-time PCR. The protein levels of calpain-2, Atg5, Atg7, Atg12 and microtubule-associated protein 1 light chain 3 (LC3) were determined by Western blot. The interaction between calpain-2 and Atg5 was investigated by co-immunoprecipitation (Co-IP). RESULTSThe proliferation of BRL-3A cells treated with DTT was significantly inhibited. The apoptosis of BRL-3A cells was significantly increased after DTT treatment for 6, 12 and 24 h as compared with 0 h group (P<0.05). The cell cycle was arrested in G₁ phase after DTT treatment (P<0.05). After DTT treatment for 6, 12 and 24 h, the mRNA expression of calpain-2 and Atg5 in the BRL-3A cells was significantly increased as compared with 0 h group (P<0.05). The protein levels of calpain-2, Atg12 and Atg7 in the cells treated with DTT for 6, 12 and 24 h were significantly higher than those in 0 h group, and the ratio of LC3-II/LC3-I was also significantly higher than that in 0 h group, while Atg5 expression was significantly lower than that in 0 h group (P<0.05). The results of Co-IP found that the anti-calpain-2 antibody precipitated Atg5 protein from the cell lysates, and the anti-Atg5 antibody also precipitated calpain-2 from the cell lysates, which confirmed the interaction between calpain-2 and Atg5. CONCLUSION Calpain-2 may participate in ERS-induced hepatocyte apoptosis by interacting with Atg5.     

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.     

mTOR kinase inhibitor CC-223 suppresses human breast cancer cell viability

AIM: To investigate the inhibitory effect of CC-223, an inhibitor of mammalian target of rapamycin (mTOR) kinase, on the viability of human breast cancer cells and its mechanism. METHODS: The inhibitory effect of CC-223 on the viability of MCF-7 cells and MDA-MB-231 cells was measured by CCK-8 assay. The cell cycle distribution of breast cancer cells was examined by flow cytometry. The expression of cell cycle-related proteins and oncoproteins c-Myc and survivin was analyzed by Western blot. RESULTS: CC-223 significantly inhibited the viability of both MCF-7 and MDA-MB-231 cells (P<0.05). CC-223 induced cell cycle arrest in both G₁ phase and G₂/M phase in the MCF-7 cells (P<0.05). However, low concentration of CC-223 treatment resulted in the accumulation of MDA-MB-231 cell cycle in the G₂/M phase, and the cell number in G₁ phase was unaffected. Treatment with CC-223 for 24 h clearly inhibited the protein levels of cyclin B1, cyclin D1 and phosphorylated cell division cycle protein 2 in the breast cancer cells (P<0.05). CC-223 suppressed the expression of c-Myc and survivin in both MCF-7 cells and MDA-MB-231 cells (P<0.05). CONCLUSION: CC-223 inhibits cell viability by blocking cell cycle progression and down-regulating expression of c-Myc and survivin in both MCF-7 and MDA-MB-231 cells.     

Crude garlic extract significantly inhibits replication of grapevine viruses

Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses.     

灌浆温度和氮肥及其互作效应对稻米贮藏蛋白组分的影响

灌浆结实期温度与氮肥施用量是影响稻米品质的两个重要生态因子,尤其是与稻米蛋白含量及米饭食味关系密切。本文以多个水稻主栽品种为材料,通迆灌浆结实期的人工控温试验、大田长期定位点的施氮处理试验和盆栽条件下的温氮两因素复合处理试验,探讨了水稻灌浆结实期温度对稻米贮藏蛋白含量与组分影响及其有别于氮肥处理效应的差异觃律,幵分析了温度与氮肥两个因素对稻米贮藏蛋白及其组分影响的交互作用特点。结果表明,高温胁迫和增施氮肥均引起水稻籽粒总蛋白及其谷蛋白组分含量(%)的显著增加,但两者对稻米醇溶蛋白影响却存在明显差别。其中,高温处理引起醇溶蛋白含量显著下降,提高稻米谷蛋白/醇溶蛋白比值,而增施氮肥引起稻米谷蛋白和醇溶蛋白含量明显增加,但对谷蛋白/醇溶蛋白比值与贮藏蛋白各亚基的组成比例影响相对较小。在高温处理下,谷蛋白的57kD前体亚基组分含量有所提高,而37kD酸性亚基和22kD碱性亚基随温度处理的差异变化却因品种而异,且高温处理对水稻籽粒蛋白绝对含量(mg grain~(–1))的影响程度也进没有其对蛋白相对含量(%)的影响明显。高氮×高温处理组合对稻米总蛋白与谷蛋白含量的影响程度显著大于单一高温或高氮处理,但在高氮水平下由高温引起稻米醇溶蛋白含量的下降幅度却小于其低氮对照,有利于稻米醇溶蛋白含量在不同温度处理下的相对稳定。