棉铃虫对菊酯类杀虫剂抗药性的神经电生理研究

本文用神经电生理方法研究了氰戊菊酯、氯菊酯对棉铃虫Helicoverpa armigera(Hubner)相对敏感(HD-S)种群和抗性(HJ-R)种群的神经毒理作用。10^-5mol/L的氰戊菊酯、10^-5mol/L的氯菊酯诱发腹神经索自发发放频率的增加和随后的神经传导阻断,10-^-5mol/l的氯菊酯抑制HD-S种群的神经兴奋,直接阻断神经传导。以兴奋时间、神经传导阻断时间、对药剂作用反应时间的个体分布频率3个参数比较两种群对杀虫剂的反应,均发现HJ-R种群比相对HD-S种群表现了2～3倍的神经不敏感机制,并且发现这种神经不敏感机制对毒理I型和Ⅱ型拟除虫菊酯同样有作用。     

高效氯氟氰菊酯对大鳞副泥鳅急性毒性和生理毒性的影响

为了估测高效氯氟氰菊酯对水生生物毒性的大小,以大鳞副泥鳅作为受试动物,研究不同浓度的高效氯氟氰菊酯对其急性毒性和生理毒性的影响。结果表明:高效氯氟氰菊酯对大鳞副泥鳅24h、48h、96h的半数致死浓度(LC50)分别为25.93μg/L、17.28μg/L、14.11μg/L,安全浓度为2.30μg/L。低浓度高效氯氟氰菊酯对大鳞副泥鳅肝脏谷丙转氨酶(GPT)和谷草转氨酶(GOT)起诱导作用,高浓度对其起抑制作用,试验组与对照组存在显著或极显著差异。研究表明,高效氯氟氰菊酯对大鳞副泥鳅有较强的毒性。     

氯吡格雷对大鼠肝药物酶CYP3A4 和CYP1A2 表达的影响

目的：氯吡格雷主要由CYP3A4 催化使其激活,CYP1A2 也参与氯吡格雷活化。关于氯吡格雷对肝微粒体酶的影响国内外 文献报道不多,因此本实验通过检测肝细胞色素氧化酶CYP3A4 和CYP1A2 的表达,探讨氯吡格雷对大鼠肝药物酶的影 响。方法：生理盐水为对照组,氯吡格雷设高、中、低三个剂量组（27,13.5,6.75mg/kg/d）,雄性健康大鼠连续灌胃给药7天,脱臼处 死,取肝组织,通过western blot法检测大鼠肝脏CYP3A4 和CYP1A2 蛋白表达情况。结果：1）、氯吡格雷抑制大鼠CYP3A4 蛋白 表达,氯吡格雷高中低剂量组分别比生理盐水组大鼠CYP3A4 蛋白表达量降低（P<0.05）;氯吡格雷低中高剂量组间进行比较,大 鼠CYP3A4 蛋白表达量呈梯度减少（P<0.05）;2）、氯吡格雷抑制大鼠CYP1A2 蛋白表达,氯吡格雷高中低剂量组分别比生理盐水 组大鼠CYP1A2 蛋白表达量降低（P<0.05）,氯吡格雷低中高剂量组间进行比较,大鼠CYP1A2 蛋白表达量呈梯度减少（P<0.05）。 结论：氯吡格雷使肝细胞色素氧化酶CYP3A4 和CYP1A2 的表达量减少,因此氯吡格雷高、中、低3 个剂量组均不同程度的抑制 大鼠肝脏CYP3A4 和CYP1A2 的表达,提示当氯吡格雷与某些主要经CYP3A4 和CYP1A2 代谢的药物合用时,发生代谢性相关 作用的可能性大。     

氯吡格雷对大鼠肝药物酶CYP3A4和CYP1A2表达的影响

目的：氯吡格雷主要由CYP3A4催化使其激活,CYPlA2也参与氯吡格雷活化。关于氯吡格雷对肝微粒体酶的影响国内外文献报道不多,因此本实验通过检测肝细胞色素氧化酶CYP3A4和CYPlA2的表达,探讨氯吡格雷对大鼠肝药物酶的影响。方法：生理盐水为对照组,氯吡格雷设高、中、低三个剂量组（27,13．5,6．75mg／kg／d）,雄性健康大鼠连续灌胃给药7天,脱臼处死,取肝组织,通过westernblot法检测大鼠肝脏CYP3A4和CYPlA2蛋白表达情况。结果：1）、氯吡格雷抑制大鼠CYP3A4蛋白表达,氯吡格雷高中低剂量组分别比生理盐水组大鼠CYP3A4蛋白表达量降低（P〈0．05）;氯吡格雷低中高剂量组间进行比较,大鼠CYP3A4蛋白表达量呈梯度减少（P〈0．05）;2）、氯吡格雷抑制大鼠CYPlA2蛋白表达,氯吡格雷高中低剂量组分别比生理盐水组大鼠CYPlA2蛋白表达量降低（P〈0．05）,氯吡格雷低中高剂量组间进行比较,大鼠CYPlA2蛋白表达量呈梯度减少（P〈0．05）。结论：氯吡格雷使肝细胞色素氧化酶CYP3A4和CYPlA2的表达量减少,因此氯吡格雷高、中、低3个剂量组均不同程度的抑制大鼠肝脏CYP3A4和CYPlA2的表达,提示当氯吡格雷与某些主要经CYP3A4和CYPlA2代谢的药物合用时,发生代谢性相关作用的可能性大。     

不同pH和温度对A型肉毒毒素结构及活性的作用

目的研究p H和温度对A型肉毒毒素结构及活性的影响。方法将A型肉毒毒素溶解于p H6.6和p H7.8的磷酸盐缓冲液(PB)中,分别采用4℃和35℃两种温度保存30天,在不同时间采用高压液相色谱(HPLC)及小鼠试验研究其结构及生物学活性的变化。结果样品1(p H6.6,4℃)的HPLC图谱30天内只呈现Ⅰ峰(复合物),毒力未见明显下降;样品2(p H7.8,4℃)则在溶解的最初便有Ⅱ峰(神经毒素)解离出来,并且随着时间的增加Ⅱ峰所占比例逐渐增大,毒力在30天内未见明显下降;样品3(p H6.6,35℃)最初只呈现Ⅰ峰,在第7天便有Ⅱ峰解离出来,第21天开始Ⅲ峰(轻链)出现并伴随着毒力的大幅下降(最初毒力的50%),30天降低至最初毒力的约40%;样品4(p H7.8,35℃)在第1天便有Ⅰ、Ⅱ、Ⅲ峰出现,伴随着毒力的大幅下降(最初毒力的50%),30天降低至最初毒力的约25%。结论采用酸性条件有利于维持A型肉毒毒素复合体的结构,碱性会破坏复合体各组分间的非共价键。不论用酸性还是碱性条件,高温度会加速复合体的解离与神经毒素二硫键的断裂,二硫键的断裂导致活性的下降。     

circSAMD4A靶向miR-141-3p对肺泡上皮细胞损伤的作用和机制

该文旨在探讨circ SAMD4A对肺炎链球菌(Streptococcus pneumoniae, SP)诱导的肺泡上皮细胞损伤的作用,并分析其分子机制。将肺泡上皮细胞分为对照组、感染组、感染+si-circSAMD4A组、感染+si-NC组、感染+mi R-141-3p组、感染+mi R-NC组、感染+si-circ SAMD4A+antimi R-NC组、感染+si-circ SAMD4A+anti-mi R-141-3p组; qRT-PCR测定circ SAMD4A和mi R-141-3p水平;酶联免疫吸附试验(ELISA)测定IL-6、IL-10水平;流式细胞术检测细胞凋亡率;蛋白质印迹法检测Cleaved-caspase3和Cleaved-caspase9蛋白水平;双荧光素酶报告实验检测circ SAMD4A和mi R-141-3p的靶向关系。SP诱导的肺泡上皮细胞中circSAMD4A表达水平、IL-6水平、细胞凋亡率、Cleaved-caspase3和Cleaved-caspase9蛋白表达水平升高, miR-141-3p和IL-10表达水平、细胞活力降低(P<0....     

电刺激尾核头部对隐神经A类和C类纤维传入诱发体感皮层电反应的影响

实验用猫,在氯醛糖麻醉、三碘季铵酚制动下,用方波电脉冲刺激隐神经,配合极化电流阻断技术分别引起隐神经Ａ和Ｃ类纤维兴奋,在对侧大脑皮层ＳⅠ区和ＳⅡ区记录平均诱发电位（分别同称为Ａ－ＳⅠＥＰ、Ａ－ＳⅡＥＰ、Ｃ－ＳⅠＥＰ、Ｃ－ＳⅡＥＰ）。用双极同心圆电极刺激尾核头部不同部位。实验结果表明,只有刺激同侧尾核头部内侧浅表部位,才对Ｃ－ＳⅠＥＰ,Ｃ－ＳⅡＥＰ,Ａ－ＳⅡＥＰ有抑制作用,提示尾核头部对大脑皮层不同躯体感觉代表区有不同的调制作用,对ＳⅠ区Ｃ类纤维传入信息有独特的抑制作用。     

不同剂量炔雌醚对小鼠器官鲜重、繁殖生理和肝肠CYP3A4酶含量的影响

为探究不同剂量炔雌醚对小鼠器官、激素和肝肠药解酶的影响,分别以0.008 mg/kg、0.04 mg/kg、0.2 mg/kg、1.0 mg/kg和5.0 mg/kg的炔雌醚油溶液连续3d灌胃小鼠,首次给药7d后解剖取材,检测其器官鲜重、雄鼠精子数量、血清中激素浓度、小肠和肝脏中CYP3 A4酶含量的变化.结果 发现:...     

高压氧联合高频rTMS对重型颅脑损伤后意识障碍患者促醒作用、神经电生理和脑损伤标志物的影响

摘要 目的：探讨高压氧联合高频重复经颅磁刺激（rTMS）对重型颅脑损伤后意识障碍（DOC）患者促醒作用、神经电生理和脑损伤标志物的影响。方法：选取2021年6月~2023年6月期间安徽中医药大学附属六安医院收治的的80例重型颅脑损伤后DOC患者。根据随机数字表法分为对照组（n=40,高压氧治疗）和观察组（n=40,高压氧联合高频rTMS治疗）。对比两组疗效、促醒作用、神经电生理和脑损伤标志物水平变化情况。结果：观察组的临床总有效率高于对照组（P<0.05）。两组治疗4周后修订的昏迷恢复量表（CRS-R）、格拉斯哥昏迷量表（GCS）评分升高,观察组高于对照组（P<0.05）。两组治疗4周后脑干听觉诱发电位（BAFP）、四肢体感诱发电位（SEP）、脑电图（EEG）分级有所改善,观察组改善效果优于对照组（P<0.05）。两组治疗4周后髓鞘碱蛋白（MBP）、神经元特异性烯醇化酶（NSE）、S100β蛋白下降,观察组低于对照组（P<0.05）。结论：高压氧联合高频rTMS治疗可有效促醒重型颅脑损伤后DOC患者,还可改善神经电生理活动,减轻脑损伤程度。     

Aβ_(25～35)和人参皂苷Rb1对鼠神经干细胞分化后GSK-3β、CDK-5和PP2A表达的影响

目的:研究大鼠神经干细胞诱导分化后GSK-3β、CDK-5和PP2A的表达以及Aβ25～35和人参皂苷Rb1的调节作用。方法:取新生SD大鼠的海马组织体外培养获得NSCs,诱导第3代的NSCs向神经细胞分化1周后,免疫荧光细胞化学染色检测活化型GSK-3β(pTyr279,216)和蛋白磷酸酯酶-2A(PP2A)的表达及Aβ25～35和人参皂苷Rb1对它们的影响;RT-PCR分析糖原合成激酶-3β(GSK-3β)、细胞周期依赖性蛋白激酶-5(CDK-5)和蛋白磷酸酯酶-2A(PP2A)的mRNA表达以及Aβ25～35和人参皂苷Rb1的影响。结果:免疫荧光细胞化学染色显示:NSCs诱导分化1周后有活化型GSK-3β(pTyr279,216)和PP2A的表达,Aβ25～35能增强GSK-3β(pTyr279,216)的表达同时抑制PP2A的表达,而人参皂苷Rb1则能逆转Aβ25～35的作用;RT-PCR检测发现:NSCs诱导分化1周后表达GSK-3β、CDK-5和PP2A的mRNA,使用Aβ25～35处理后GSK-3β、CDK-5的表达增强而PP2A的表达减弱,用人参皂苷Rb1预处理神经干细胞后Aβ25～35的作用受到抑制。结论:体外培养的神经干细胞分化后表达GSK-3β、CDK-5和PP2A,并受Aβ25～35和人参皂苷Rb1的调节,提示在体外由神经干细胞分化的细胞具备正常神经细胞的蛋白磷酸化调节系统。     

Development and characterization of microsatellite markers in the Colorado potato beetle,Leptinotarsa decemlineata

The characterization of 11 Leptinotarsa decemlineata microsatellite loci isolated using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol is reported. Three to 15 alleles per locus were detected in 54 beetles collected from four populations of L. decemlineata. The mean number of alleles was 7.4 ± 3.2 and the level of expected heterozygosity ranged from 0.451 to 0.798. The total exclusionary probabilities using these loci for the first and the second parent were 0.982 and 0.999, respectively. These are the first microsatellite loci characterized from the Colorado potato beetles that can be used for estimating genetic diversity, population structure and parentage analysis.     

Ecdysteroids and adult diapause in the Colorado potato beetle, Leptinotarsa decemlineata

ABSTRACT. The difference in ecdysteroid activity in short-day (10-h photo-phase) and long-day (16-h photophase) Leptinotarsa decemlineata Say adults was characterized by high performance liquid chromatography, which revealed that for short-day beetles ecdysterone and ecdysone are the major constituents of the free ecdysteroids, whereas in long-day beetles products co-migrating with 2-deoxyecdysone and 2-deoxyecdysterone are just as abundant. Ecdysteroids were detected (15 ng/ml) only in the haemolymph of females. Ligation experiments showed that the induction of the diapause syndrome by inactivation of the corpora allata or the brain not only resulted in the formation of the typical diapause proteins but also in an increase of the ecdysteroid activity. It is suggested that adult diapause in L.decemlineata is regulated by a multifactorial system.     

Biological control potential of Turkish entomopathogenic nematodes against the Colorado potato beetle,Leptinotarsa decemlineata

Three Turkish isolates of the entomopathogenic nematodes Steinernema carpocapsae, Steinernema feltiae and Heterorhabditis bacteriophora were evaluated under laboratory conditions (in different temperatures and doses) for their biocontrol efficiency against last instar Leptinotarsa decemlineata. Herein, the effects of infective juveniles in aqueous suspension against L. decemlineata were evaluated. S. feltiae appeared to be the most pathogenic nematode among the tested species.     

Genetic relationships of introduced Colorado potato beetle Leptinotarsa decemlineata populations in Xinjiang,China

The Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), is an infamous invasive species worldwide that aggressively attacks potato and other Solanaceae crops. CPB was first found in China in 1993 and has since spread across 2.77 × 10⁵ km² in Xinjiang Uygur Autonomous Region. To better understand genetic variation and migration patterns, we used seven polymorphic microsatellite loci to elucidate the genetic relationships and gene flow among 10 CPB populations across Xinjiang. (i) Overall low levels of genetic diversity were detected on the entire population in Xinjiang but most of the diversity was retained among populations during invasion. (ii) The mean pairwise F_ST was low (0.071 ± 0.043) among populations. The genetic differentiation was little (pairwise F_ST 0.038 ± 0.016) between the five interior populations (Wusu, Urumqi, Jimsar, Qitai and Mulei) and Tacheng population. The six populations might come from the same genetic group via Bayesian clustering and were closely related on a neighbor‐joining tree. Combining the history data, the five interior populations may have originated from Tacheng. (iii) Gene flow was frequent, especially among the five interior populations. Individuals from the interior populations could be assigned to Tacheng at higher probabilities (means 0.518 ± 0.127) than vice versa (means 0.328 ± 0.074), suggesting that the beetle population has spread from the border to the interior in Xinjiang.     

Effects of four Bacillus spp. of soil origin on the Colorado potato beetle Leptinotarsa decemlineata (Say)

Four species of Bacillus were isolated from soil in an effort to find safe, effective and alternative biological control agents against plant pests. These bacteria were identified as Bacillus pumilus, Bacillus sphaericus, Bacillus megaterium and Bacillus cereus on the basis of fatty acid methyl ester analysis and carbon utilization profiles by using Microbial Identification and Biolog Microplate Systems. Laboratory experiments carried out to determine the insecticidal activities of these isolates showed that B. pumilus caused 95.7 and 26.7% mortality and B. sphaericus caused 74.5 and 23.3% mortality of Leptinotarsa decemlineata larvae and adults, respectively. B. cereus and B. megaterium showed 51.1 and 29.7%, respectively, of L. decemlineata larvae. This study presents at least two Turkish isolates from the genus Bacillus showing high insecticidal activity against L. decemlineata.     

Host-plant discrimination and evolution of feeding preference in the Colorado potato beetle Leptinotarsa decemlineata

ABSTRACT. The behaviour of newly emerged adult Colorado potato beetles on preferred hosts follows a stereotyped pattern of sampling, feeding, grooming and rest. Reduced meal sizes on less-preferred hosts is accompanied by increased sampling and frequent interruptions in feeding. A systematic increase in pre-ingestive sampling on less-preferred foodplants indicates that beetles discriminate among closely related species within the Solanaceae. This ability may depend primarily on stimuli perceived at, and near, the leaf surface. Three geographic populations of beetles have adapted to different local host plants, but have not lost their preference for feeding on an ancestral host species. Host shifts by oligophagous insects to related plant species may evolve through selection for feeding generalists in isolated populations, and may not require genetic changes affecting the perception of a particular novel host.     

Sequencing and characterization of a cDNA encoding a ferritin subunit of Colorado potato beetle, Leptinotarsa decemlineata

A differentially expressed cDNA fragment (P311) from Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), was identified by restriction fragment differential display-polymerase chain reaction (RFDD-PCR) technique, and showed a strong similarity to ferritin heavy chain subunits of other organisms. Based on P311, we constructed specific primers and obtained a 840-bp cDNA fragment spanning the open reading frame of CPB ferritin subunit using the rapid amplification of cDNA ends (RACE) technique. The sequence encodes 213 amino acid residues, including a 19 amino acid signal peptide. The sequence has a conserved cysteine in the N-terminus and has the seven conserved residues that comprise the ferroxidase center, which is the feature of heavy chain ferritins of vertebrates. The CPB ferritin subunit has high amino acid sequence identity with the Apriona germari (69.3%), Galleria mellonela (54.5%), Manduca sexta (54.0%), Drosophila melanogaster (53.2%), Calpodes ethlius (51.4%), and Nilaparvata lugens (47.6%) but lower identity with the Anopheles gambiae (38.7%) and Aedes aegypti (37.8%). Using Northern blot analysis, the subunit mRNA was identified from fat body and midgut of 4th instars with much higher mRNA levels found in midgut than that in fat body (2.5-fold). Nevertheless, only the levels of mRNA in fat body was induced by dexamethasone (1.5-fold).     

Effect of diet and feeding history on flight of Colorado potato beetle, Leptinotarsa decemlineata

We evaluated the hypothesis that Colorado potato beetle (Leptinotarsa decemlineata Say) (CPB) flight frequency is related to diet, and that it changes with duration of food unavailability or exposure to poor quality food by exposing adult overwintered and summer CPB populations to an acceptable host plant (conventional foliage), a poor host (insect resistant transgenic foliage expressing Bacillus thuringiensis tenebrionis[Btt] Cry3a toxin) and no host. Exposure to poor host and no host treatments (with or without water) decreased mean daily flight frequencies and the overall number of overwintered CPB flying, but increased the mean daily flight frequency and number of summer population CPB that flew. Overwintered CPB did not react to an absence of plants at emergence whereas summer CPB increased mean daily flight frequencies when plants and water were not available. The flight response to insect resistant foliage was similar to that for starvation treatments in both populations indicating that flight may not be triggered by Btt toxins but by starvation brought on by feeding on poor quality food. Flight was observed in all treatments for the duration of the test with two exceptions; overwintered beetles fed insect resistant foliage ceased flying after day 17 and summer beetles starved without water ceased after day 8 of a 29‐day study.     

Body movements and their role as triggers of heartbeats in pupae of the Colorado potato beetle Leptinotarsa decemlineata

Rhythmic body movements and their role as triggers of intermittent heartbeats were studied in pupae of Leptinotarsa decemlineata Say. Heartbeats and body movements were recorded simultaneously by means of an optical method (infra‐red cardiography) combined with respirometry. IR‐cardiography allows heartbeats and body movements to be distinguished on the basis of their different rates (40–80 min^?1 and 4–8 min^?1, respectively) and amplitude. In the mid‐interecdysial period, abdominal movements in the pupae were always accompanied by heart activity beginning after the first 5–12 abdominal strokes. Simultaneous periods of abdominal movements and heartbeats lasted 2–5 min, while the intervening pause ranged from 40 to 72 min at 24°C. Experiments of forced heart activity showed that a slight external tactile stimulus (prodding once with a single hair), applied prior to an expected heartbeat bout, evoked abdominal movements followed soon afterwards by contractions of the dorsal vessel. Repeated prodding with a hair evoked body rotating movements (1–3 strokes) at any time with heartbeats starting at the first movement. We surmise that rhythmic body movements play an active role in the triggering of heart activity in pupae according to the principle of cardiac reflex response. This is a possible mechanism for synchronizing periods of heart activity with periods of rhythmic body movements. Haemolymph circulation in pupae is favoured when heartbeat bouts are accompanied by active body movements.