Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.     

Binding Sites for Ca2+-Channel Effectors and Ryanodine in Periplaneta americana—Possible Targets for New Insecticides

The calcium channel and the ‘calcium release channel’ of muscle membrane of the cockroach Periplaneta americana have been characterized. Biological assays with calcium channel blockers and ryanodine on different insects and acari revealed pronounced insecticidal effects with ryanodine, but not with calcium channel blockers, at concentrations between 0·1 and 300 μg ml⁻¹. Skeletal muscle membranes derived either from the tubular network or from the sarcoplasmatic reticulum of P. americana were characterized with respect to the binding of the dihydropyridine (DHP) [³H]isradipine (PN 200-110), the phenyl-alkylamine [³H]verapamil and the alkaloid [³H]ryanodine. Preliminary binding studies with the benzothiazepine [³H]diltiazem suggest a low-affinity binding site with a IC₅₀ value of 3·3 μM . All binding sites tested were sensitive to treatment with proteinase K. Optimal conditions for binding of the radioligand ryanodine revealed the highest specific binding at pH 8 and at calcium chloride concentrations between 100 and 500 μM . EGTA at 10 μM abolished 95% of the ryanodine binding. Binding studies with calcium channel binding sites revealed a pronounced effect of low Ca²⁺ concentrations on specific isradipine binding, whereas verapamil and diltiazem binding were only reduced by the presence of 200 μM EGTA. With respect to high Ca²⁺ concentrations, specific binding of diltiazem, isradipine and verapamil was reduced by 73, 40 and 20%, respectively, at 5 mM Ca²⁺. Radioligand binding experiments showed high-affinity binding sites for ryanodine and isradipine. K_D values of 0·95 nM (B_max=550 fmol mg⁻¹ protein) and 0·75 nM (B_max=213 fmol mg⁻¹ protein) were determined respectively. A lower-affinity binding site was identified in binding studies with verapamil (K_D=7·4 nM and B_max=27 fmol mg⁻¹ protein). [³H]isradipine displacement studies with several dihydropyridines revealed the following ranking of affinity: nitrendipine>isradipine>Bay K8664≪nicardipine. Displacement of [³H]verapamil binding by effectors of the phenylalkylamine binding site showed that bepridil and S(-)verapamil had the highest affinities of the compounds tested followed by (±)verapamil, nor-methylverapamil and R(+)verapamil.     

油梨嫁接苗分级标准探讨

通过逐步聚类分析方法,初步探讨了油梨嫁接苗的分级标准。分析得出以苗高和茎粗为油梨嫁接苗的分级质量指标,将油梨嫁接苗分为三级,即Ⅰ级苗：苗高≥74cm,茎粗≥1.13cm;Ⅱ级苗：74cm＞苗高≥66 cm,1.13cm＞茎粗≥0.93cm;Ⅲ级苗：苗高＜66cm,茎粗＜0.93cm。     

美国豆芋花总皂苷制备及其α-葡萄糖苷酶抑制活性

为高效利用美国豆芋花皂苷类化合物,以皂苷得率和纯度为指标,研究美国豆芋花总皂苷的提取工艺及大孔树脂纯化工艺,并对制备的美国豆芋花总皂苷的抗氧化及α-葡萄糖苷酶活性抑制能力进行研究。结果表明,美国豆芋花总皂苷的提取工艺为提取溶剂90%乙醇,料液比1∶20,温度70℃,提取时间2 h。得到的提取物石油醚脱脂后用水饱和正丁醇萃取,D-101大孔树脂吸附柱纯化的工艺为上样皂苷质量浓度1 mg·m L~(-1),上样量40 m L·g~(-1)树脂,洗脱剂为80%乙醇(pH值6),洗脱剂用量4 BV(树脂柱体积)。制备的美国豆芋花总皂苷样得率4.24%,纯度达65.35%,比粗提物(纯度15.27%)提高了约3倍;抗氧化性能(清除DPPH和ABTS自由基IC50值分别为51.08、29.24μg·m L~(-1))和α-葡萄糖苷酶活性抑制能力(IC50值为83.62μg·m L~(-1))也均优于粗提物(清除DPPH、ABTS自由基IC50值分别为286.51、69.92μg·m L~(-1),α-葡萄糖苷酶的抑制浓度(IC50)值为1 043.57μg·m L~(-1)),表明该工艺可用于高活性美国豆芋花总皂苷的富集和纯化。此外,美国豆芋花总皂苷抑制α-葡萄糖苷酶活性的抑制类型为竞争型抑制,抑制常数为49.68μg·m L~(-1)。本研究结果为开发具有降血糖功能的医药中间体或功能性食品提供了新资源和实践基础。     

美洲大蠊提取物对四氧嘧啶性糖尿病小鼠作用研究

[目的]研究美洲大蠊(MZDL)提取物对四氧嘧啶性糖尿病小鼠血糖的影响。[方法]四氧嘧啶诱发糖尿病小鼠后,随机分为模型组(MC),二甲双胍组(250 mg/kg),美洲大蠊提取物低剂量(25 mg/kg)、中剂量(50 mg/kg)、高剂量(100 mg/kg)5个组,模型组给予同体积生理盐水,分别于试验第7、14、212、8天各测定小鼠空腹血糖和体重,第25～28天每天记录进食饮水量。28 d后取血测血清中一氧化氮合酶(NOS)含量。[结果]3个剂量的美洲大蠊提取物均能减缓糖尿病小鼠消瘦和多饮多食的症状。在试验中中、高剂量组糖尿病小鼠血糖降低(P<0.05)。MZDL中、高剂量组NOS含量低于MG(P<0.01)。[结论]美洲大蠊提取物对四氧嘧啶致糖尿病小鼠具有一定的保护作用,其机制可能与其降低一氧化氮合酶水平有关。     

Interaction of dinotefuran and its analogues with nicotinic acetylcholine receptors of cockroach nerve cords

To investigate the action of dinotefuran (MTI-446, 1-methyl-2-nitro-3-(tetrahydro-3-furylmethyl)guanidine), a recently developed insecticide, on insect nicotinic acetylcholine receptors (nAChRs), we determined the potencies of the compound and 15 analogues in inhibiting the specific binding of [3H]epibatidine (EPI), a nAChR agonist, and [3H]alpha-bungarotoxin (alpha-BGT), a competitive nAChR antagonist, to the nerve cord membranes of American cockroaches (Periplaneta americana). Racemic dinotefuran inhibited [3H]EPI binding with an IC50 of 890 nM and [3H]alpha-BGT binding with an IC50 of 36.1 microM. Scatchard analysis indicated that the dinotefuran inhibition of [3H]EPI binding was a competitive one. Slight structural modification caused a drastic reduction in potency; only four analogues were found to be equipotent to or more potent than dinotefuran. Chloropyridinyl and chlorothiazolyl neonicotinoid insecticides displayed two or three orders of magnitude higher potency than dinotefuran. There was a good correlation between the IC50 values of tested compounds obtained with [3H]EPI and those obtained with [3H]alpha-BGT. A better correlation was observed between 3-h knockdown activities (KD50) against German cockroaches (Blattella germanica) and IC50 values obtained from [3H]EPI assays than between 24-h lethal activities (LD50) and IC50 values. While the results indicate that dinotefuran and its analogues interact with the ACh-binding site in cockroach nAChRs, it remains to be elucidated why they displayed lower potencies than those expected based on their insecticidal activities.     

Effect of pre-incubation humidity and temperature treatment on the in vitro germination of avocado pollen grains

Optimum in vitro germination of pollen grain of the avocado cultivars Fuerte, Nabal, Ettinger, Bacon and Zutano occurred at 25 °C. However, there were significant differences between cultivars in percentage germination and relative humidity (RH) requirements for optimum pollen grain growth. The most sensitive cultivar to relative humidity was Fuerte, in which the germination of pollen grain rose from 11.4%, at 40% RH, to about 50%, after one hour at 100% RH. The germination% of Nabal pollen grain was already high at 40% RH and was not increased by higher relative humidity. Increased relative humidity also helped to sustain the viability of avocado pollen. At 30 °C and 5% RH the pollen grains of Fuerte quickly lost its ability to germinate, at 40% RH for 1 hour, germination was reduced spectacularly compared to pollen kept in saturated with moisture environment where it was not affected the first 24 hours. The effects of temperature and relative humidity on fruit-set and yield of avocado are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cellular Defence Reaction of American Cockroach , Periplaneta americana to Metarhizium anisopliae Isolate CQMa102

Injection of conidia solution of Metarhizium anisopliae isolate CQMa102 into the hemocoel of Periplaneta americana caused significant changes of the total hemocytes of the cockroach. P. americana eliminated large amount of the spores efficiently by phagocytosis and nodulation.The experiment shows that the immune system of the cockroach was hardly injured after injected with high dose spores, and the higher dose of spores was injected,the higher mortality of the insect was. However, low dose injection strengthened the immune ability of the insect in certain degree. The saline slightly induced the immune reaction of the insect.     

1-MCP对青脆李亚低温贮藏期间生理生化及品质的影响

以四川茂县青脆李为试材,研究1-甲基环丙烯(1-MCP)对亚低温(8±0.5℃)条件下青脆李的生理生化及其贮藏期间果实品质变化的影响。结果表明:1-MCP处理可显著抑制青脆李果实的呼吸作用和乙烯的释放速率,并降低两者的峰值;可保持果实细胞膜的完整性,降低PPO活性和酚类物质的消耗,减少褐变。其中以1.0μL.L-1浓度的1-MCP处理能够更有效地保持果实的质地与风味,亚低温条件下贮藏28 d,褐变指数为0,腐烂指数为5.3,显著好于对照和其他浓度的1-MCP处理,延长果实保鲜期达到20 d。     

室内人工繁殖美洲大蠊的产卵习性

美洲大蠊从4月份开始活动,产卵高峰期发生在6月下旬至9月中旬,它喜欢于角落产卵,并有自食其卵的习性.用标准饲料饲养美洲大蠊,并提供足够的水,增大养虫笼容积,增设阻隔层,可降低美洲大蠊食卵率.