突变肠杆菌株NRS-1中5个草甘膦逆境应答基因的克隆与功能研究

【目的】微生物对草甘膦的抗性受复杂的遗传体系调控,涉及靶基因和大量相关调控基因。对肠杆菌属细菌NRS-1突变菌株在高浓度草甘膦逆境下的5个重要差异表达基因进行功能研究,以期深入了解非靶标基因在抗草甘膦微生物的作用特点,为发掘优异基因资源,服务抗草甘膦转基因生物育种提供参考。【方法】NRS-1的差异表达基因可能在蛋白质合成、代谢、细胞膜等水平发挥作用,保护细胞免受高浓度草甘膦逆境,因此分别选取易位酶延伸因子fus A、丁二酸脱氢酶sdh A、胸苷磷酸化酶deo A、鸟氨酸氨甲酰转移酶arg F、周质蛋白osm Y进行克隆,采用大肠杆菌原核表达、转化拟南芥实验研究其功能,并通过细菌双杂及KEGG pathway分析基因间互作特点。【结果】在明确5个基因结构特点基础上,通过大肠杆菌原核表达及转基因拟南芥鉴定,发现这5个基因及草甘膦的靶基因5-烯醇式丙酮莽草酸-3-磷酸合成酶基因aro A对提高2种生物的草甘膦耐性均有不同程度的作用,其中arg F、deo A的抗性较好,与aro A相当,表明在应对草甘膦逆境时,芳香族、含有胸腺嘧啶氨基酸及精氨酸的合成代谢通路可能起重要作用;利用基因互作与KEGG分析发现5个基因与靶基因aro A间形成复杂的调控网络,但无直接的蛋白互作。【结论】NRS-1的5个差异表达基因对草甘膦逆境具有抗性,arg F、deo A优于其他3个基因,其与靶基因aro A间表现复杂的基因互作关系。     

不同时间高压氧治疗对颅脑损伤患者认知功能障碍的影响

目的:探讨不同时间高压氧治疗对颅脑损伤患者认知功能障碍的影响。方法:选择了2006年5月-2015年5月在我院接受高压氧治疗的500例颅脑损伤手术或非手术患者,根据患者开始采取高压氧治疗时间的不同分为A(入院后7d内)、B(7-30 d)、C(30 d后)三组,通过相应的治疗比较三组患者认知功能评分、FIM评分及治疗效果。结果:高压氧治疗后,三组患者较治疗前都有了不同程度上的改善,但A组在定向能力、专注能力、理解能力、复述能力、结构组织能力、记忆能力、计算能力以及自我照顾、转移、行走、交流、社会认知较B组、C组改善显著(P0.05),而B组与C组间认知功能各项评分、FIM评分比较差异无统计学意义(P0.05);A组患者治疗后总有效率为90.63%,显著优于B组73.89%的总有效率及C组70.00%的总有效率(P0.05),而B组与C组比较差异无统计学意义(P0.05)。结论:颅脑伤患者尽早的接受高压氧治疗其认知功能改善越明显,治疗效果越好。     

奥美拉唑不同给药方式治疗新生儿应激性溃疡的效果分析

目的:探讨奥美拉唑不同给药方式治疗新生儿应激性溃疡的临床效果及安全性。方法:选取2014年1月～2018年6月我院收治的应激性溃疡新生儿61例,根据随机数字表法分为两组,对照组患儿给予奥美拉唑0.6 mg/kg胃管注入治疗,观察组患儿给予奥美拉唑2 mg/kg持续24 h微泵静脉滴注。比较两组患儿的临床治疗效果、临床症状缓解时间、止血时间、胃肠喂养开始时间、p H值和不良反应的发生情况。结果:治疗后,两组患儿的临床总有效率比较无统计学差异(P0.05);观察组患儿临床症状消失时间和止血时间均显著短于对照组(P0.05),喂养开始时间≤24 h的患儿比例高于对照组,喂养开始时间≥72 h的患儿比例显著低于对照组(P0.05),pH值显著高于对照组(P0.05)。两组患儿治疗期间均未发生恶心、呕吐、便秘、皮疹等不良反应,且肝肾功能未受明显影响。结论:2 mg/kg奥美拉唑持续24 h微泵静脉滴注与0.6 mg/kg胃管注入治疗的临床总有效率相当,但微泵静脉滴注可显著提高患儿的pH值,缩短临床症状恢复时间、止血时间和胃肠喂养开始时间,且安全性高。     

混合甲醛固定液固定大肠癌淋巴结标本的最佳免疫组化效果研究

目的:探讨混合甲醛固定液固定大肠癌淋巴结标本的最佳免疫组化效果。方法:采用不同pH值(6.0、7.0、8.0)的混合甲醛固定液对39枚大肠癌淋巴结标本进行不同时间(6 h、6 h-12 h、1 d-7 d)的固定处理。以细胞角蛋白20(CK20)为目标抗原,运用OIympusdp 70图像采集分析仪抽选出混合甲醛固定液最佳免疫组化染色的pH值及固定时间。结果:经pH值为7.0混合甲醛固定液处理后,阳性率为92.31%,高于经pH值为6.0、8.0的混合甲醛固定液处理后的76.92%、74.36%,且经pH值为7.0、8.0处理后的阳性率比较有统计差异(P0.05)。混合甲醛固定液的固定时间在6 h-12 h时的阳性率为94.87%,高于固定时间为6 h、1 d-7 d处理的30.77%、76.92%(P0.05)。结论:对于大肠癌淋巴结标本,以CK20为目标抗原,选择pH值为7.0的混合甲醛固定液固定6 h-12 h能够得到质量较佳的免疫组化染色效果。     

不同剂量瑞舒伐他汀钙治疗脑梗死患者的临床疗效及机制研究

目的:探讨不同剂量瑞舒伐他汀钙治疗脑梗死患者的临床疗效,并分析其疗效机制。方法:将2015年2月至2018年2月医院诊治的脑梗死患者84例按随机数字表法分为观察组(42例)及对照组(42例)。所有患者入院后均给予降血压、降血糖、稳定颅内压以及改善微循环等基本治疗,在此基础上,对照组口服瑞舒伐他汀钙10 mg/d,观察组口服瑞舒伐他汀钙20 mg/d,两组疗程均为14 d。比较两组疗效、治疗前后美国国立卫生研究所卒中量表(NIHSS)评分以及日常生活能力量表(Barthel指数)变化,并比较两组血清白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、高迁移率族蛋白-1(HMGB1)和超敏C反应蛋白(hs-CRP)水平,分析血清HMGB1、IL-6、IL-8和hs-CRP之间的相关性,记录两组患者治疗过程中不良反应发生情况。结果:观察组治疗总有效率为95.24%(40/42),明显高于对照组的80.95%(34/42)(P0.05)。两组治疗后NIHSS评分均低于治疗前,Barthel指数高于治疗前,同时,观察组NIHSS评分降低程度大于对照组,Barthel指数升高程度大于对照组(P0.05)。治疗后两组患者血清IL-6、IL-8、HMGB1、hs-CRP明显降低,且观察组血清IL-6、IL-8、HMGB1、hs-CRP明显低于对照组(P0.05)。Person相关性分析表明患者血清HMGB1与IL-6、IL-8、hs-CRP呈正相关(r=0.306,0.428,0.367,均P0.05),IL-6与IL-8、hs-CRP呈正相关(r=0.327,0.385,P0.05),IL-8与hs-CRP亦呈正相关(r=0.430,P0.05)。治疗期间两组不良反应发生率比较无统计学差异(P0.05)。结论:高剂量瑞舒伐他汀钙治疗脑梗死能够促进神经功能恢复,提高日常生活能力,临床疗效显著,其机制可能与降低脑梗死急性期炎症反应有关。     

Correlation analysis of Normalized Different Vegetation Index (NDVI) difference series and climate variables in the Xilingole steppe, China from 1983 to 1999

There is a crucial need in the study of global change to understand how terrestrial ecosystems respond to the climate system. It has been demonstrated by many researches that Normalized Different Vegetation Index (NDVI) time series from remotely sensed data, which provide effective information of vegetation conditions on a large scale with highly temporal resolution, have a good relation with meteorological factors. However, few of these studies have taken the cumulative property of NDVI time series into account. In this study, NDVI difference series were proposed to replace the original NDVI time series with NDVI difference series to reappraise the relationship between NDVI and meteorological factors. As a proxy of the vegetation growing process, NDVI difference represents net primary productivity of vegetation at a certain time interval under an environment controlled by certain climatic conditions and other factors. This data replacement is helpful to eliminate the cumulative effect that exist in original NDVI time series, and thus is more appropriate to understand how climate system affects vegetation growth in a short time scale. By using the correlation analysis method, we studied the relationship between NOAA/AVHRR ten-day NDVI difference series and corresponding meteorological data from 1983 to 1999 from 11 meteorological stations located in the Xilingole steppe in Inner Mongolia. The results show that: (1) meteorological factors are found to be more significantly correlation with NDVI difference at the biomass-rising phase than that at the falling phase; (2) the relationship between NDVI difference and climate variables varies with vegetation types and vegetation communities. In a typical steppe dominated by Leymus chinensis, temperature has higher correlation with NDVI difference than precipitation does, and in a typical steppe dominated by Stipa krylovii, the correlation between temperature and NDVI difference is lower than that between precipitation and NDVI difference. In a typical steppe dominated by Stipa grandis, there is no significant difference between the two correlations. Precipitation is the key factor influencing vegetation growth in a desert steppe, and temperature has poor correlation with NDVI difference; (3) the response of NDVI difference to precipitation is fast and almost simultaneous both in a typical steppe and desert steppe, however, mean temperature exhibits a time-lag effect especially in the desert steppe and some typical steppe dominated by Stipa krylovii; (4) the relationship between NDVI difference and temperature is becoming stronger with global warming. __________ Translated from Acta Phytoecologica Sinica, 2005, 29(5): 753–765 [译自: 植物生态学报]     

不同年龄大熊猫肠道可培养芽孢杆菌的多样性及部分功能特性分析

【背景】大熊猫的肠道内微生物类群丰富,种群结构与宿主的年龄、生存环境、季节变化等因素有关,其中年龄是影响肠道菌群组成的重要因素之一。【目的】以不同年龄阶段的大熊猫粪便为研究对象,旨在了解不同年龄大熊猫肠道内芽孢杆菌的多样性,探究大熊猫肠道芽孢杆菌种类与年龄段之间的关系,并为优良益生菌剂的开发提供菌种资源。【方法】用稀释涂布法分离大熊猫粪便中的芽孢杆菌,对分离的菌株进行BOXA1R-PCR、16S rRNA基因系统发育及主成分分析,揭示大熊猫肠道中可培养芽孢杆菌的多样性;采用对峙生长法和药敏纸片琼脂扩散法分别检测菌株的抗菌能力和药敏性。【结果】从大熊猫粪便中共分离出90株芽孢杆菌,基于BOXA1R-PCR分析菌株的遗传多样性并从中选取41株代表菌株,经16Sr RNA基因测序分析后,结果显示归属于枯草芽孢杆菌(Bacillus subtilis)、萎缩芽孢杆菌(Bacillus atrophaeus)、贝莱斯芽孢杆菌(Bacillus velezensis)、甲基营养型芽孢杆菌(Bacillusmethylotrophicus)、解淀粉芽孢杆菌(Bacillusamyloliquefaciens)和短小芽孢杆菌(Bacillus pumilus)6个种;主成分分析结果表明大熊猫肠道芽孢杆菌种群组成与年龄存在一定的相关性。所有的供试菌株都具有纤维素降解潜力,大部分菌株对病原菌具有不同程度的抑制作用;除对青霉素具有耐药性外,供试菌株对常见的抗生素耐受性低。【结论】年龄是影响大熊猫肠道芽孢杆菌分布的重要因素,成年大熊猫肠道芽孢杆菌种类多样性最丰富,这为大熊猫益生菌制剂的开发提供了菌种资源。     

Inducible cyclooxygenase released prostaglandin mediates immunosuppression in acute phase of experimental Trypanosoma cruzi infection

We investigated the possible role of prostaglandins produced by COX-2 in the immunosuppression observed during Trypanosoma cruzi infection. Con-A-stimulated splenocytes isolated from mice on days 5, 10, and 15 of infection released large amounts of PGE2 and this release was inhibited by the treatment of animals with sodium salicylate or meloxicam. The treatment of the animals with these drugs enhanced the release of IL-2 by splenocytes from T. cruzi-infected animals and significantly reduced the blood parasitemia and delayed the mortality of the infected mice. Furthermore, the release of TNF-alpha, IFN-gamma, IL-4, and IL-10 by Con-A-stimulated splenocytes obtained from infected mice on days 5, 10, and 15 of the infection was significantly inhibited by treatment of the animals with salicylate or meloxicam. In conclusion, the results suggest that the prostaglandins produced mainly by COX-2 mediate the immunosuppression observed in the acute phase of T. cruzi infection.