急性臭氧暴露对大鼠肺部细胞遗传毒性的影响*

目的: 探讨不同浓度臭氧急性暴露对大鼠肺部细胞的遗传毒性的影响。方法: 36只wistar大鼠随机分为对照组(过滤空气暴露)、臭氧暴露组(0.12 ppm、0.5 ppm、1.0 ppm、2.0 ppm、4.0 ppm)共6组,每组6只。以不同浓度的臭氧对大鼠进行动态染毒4 h后,取肺组织并分离单细胞,采用酶联免疫吸附法检测8-羟基脱氧鸟苷(8-OHdG),利用彗星实验、微核试验和DNA-蛋白质交联实验进行DNA和染色体损伤分析。结果: 与对照组相比,肺组织中8-OHdG含量从臭氧暴露浓度为0.12 ppm起即显著增加,在0.5 ppm时达到最高值。随着臭氧暴露浓度升高,彗星拖尾率逐渐上升,且存在明显的剂量-效应关系;DNA-蛋白质交联率有先升高后下降的趋势,且在2.0 ppm时达到最大值;而肺部细胞微核率尽管呈现出上升趋势,但与对照组相比无显著性差异。结论: 急性臭氧暴露在较低浓度(0.12 ppm)时即可导致大鼠肺部细胞的DNA损伤;而在较高浓度(4 ppm)时却未见显著的染色体损伤。     

+Gz反复暴露对大鼠主动脉和心脏组织中HO-1、p21及LCAD表达的影响

目的：观察＋Gz反复暴露对大鼠心脏及主动脉组织HO-1、p21和LCAD基因表达的影响。方法：提取＋Gz反复暴露后1h组及对照组大鼠心脏、主动脉组织总RNA,经半定量RT—PCR检测其mRNA水平。结果：＋Gz反复暴露后心脏组织HO-1、p21和LCAD mRNA水平明显升高,主动脉组织中LCAD mRNA水平上调。结论：在＋Gz反复暴露状态下,HO-1是。脏组织中一个早期重要的抗过氧化损伤,维持正常功能的保护性因子。     

富含血小板血浆和脐带干细胞联用对糖尿病足模型lncRNA表达谱的影响

目的:探讨富含血小板血浆(PRP)和脐带干细胞(UBC-MSCs)联用对糖尿病足(DF)模型lncRNA表达谱的影响。方法:采用SPF级SD大鼠(20只)建立糖尿病足模型,随机分为4组,每组5只大鼠。A组注射PBS于DF动物溃疡处,B组采用人PRP注射于DF动物溃疡处,C组采用人UBC-MSCs注射于DF动物溃疡处,D组采用人UBC-MSCs+PRP注射于DF动物溃疡处。治疗14d后,记录大鼠的血糖水平与血清CRP、TNF-α、VEGF相对表达,采用arraystar公司12×135K芯片测定lncRNA的表达谱。结果:治疗14d后,B、C、D组均较A组显著降低,D组显著低于B、C组,组间差异有统计学意义(P0.05);D组大鼠的血清CRP、TNF-α相对表达较A、B、C组显著降低,血清VEGF相对表达较A、B、C组显著升高,差异都有统计学意义(P0.05)。B组与A组比较存在1563个显著差异的IncRNA,C组与A组比较存在1982个显著差异的IncRNA,D组与A组比较存在2854个显著差异的IncRNA。将靶基因带入差异m RNA数据,筛选得到与DF相关的靶基因的lncRNA有6条,具体为CHI3L1、FA2H、CAMTA1、DOCK、STRADB和TNFRSF1A (P1101039、BF74369、Uc001aoh、N385370、AF116618和HMlincRNA1336)。结论:UBC-MSCs和PRP联合可能通过改变lncRNA的表达,影响炎症因子、血管生成因子等的表达,进而发挥降血糖和治疗DF的目的。     

慢性肾衰病人血浆MMS对大鼠心脏活动能力的影响

血浆中分子物质 (ＭＭＳ)是分子量在 30 0～ 5 0 0 0Ｄａｌｔｏｎ之间的一类多肽。有实验证明 :肾衰患者血浆ＭＭＳ在体外可抑制大鼠膈肌切片、肾皮质、大脑皮质和人类红细胞对葡萄糖的摄取 ,可抑制腺苷酸环化酶、Ｎａ Ｋ ＡＴＰ酶的活性。ＭＭＳ影响机体多种酶的活性及膜的物质转运 ,势必会导致心肌的电生理特性及心肌舒缩能力的变化 ,甚至出现尿毒症性心肌病。本实验目的在于观察ＭＭＳ对心肌舒缩能力的影响 ,为肾衰患者的心功能改变及治疗提供理论基础。1　材料与方法(1)ＭＭＳ的分离与测定　取正常人和慢性肾衰 (尿毒症期 )患者空腹血 …     

臭氧暴露对大豆叶片群体光合作用及产量的影响

臭氧（O₃）已经成为我国许多大中城市夏季的首要污染物,其具有较强的植物毒性,严重威胁农业安全。O₃污染常发生于高温晴天,具有间歇性和累积性的特点,但现有研究多集中于在叶片尺度上探究长期O₃暴露对植物生理过程的影响,而间歇性暴露对植物整株生长和光合生理特性的影响鲜有报道。以大豆为实验对象,依托开顶式气室（OTC）进行间歇性臭氧暴露,探究大豆叶片群体光合作用及产量对间歇性O₃暴露的响应。结果发现（1）间歇性O₃暴露具有累积性和恢复性,在低O₃暴露剂量（AOT40≤2.47μL L^-1 h^-1）处理下,大豆植株的净光合速率降低,但与对照组无显著差异。当AOT40较高时（AOT40≤5.35μL L^-1 h^-1）,大豆植株的净光合速率显著降低,而随着O₃胁迫的消失,大豆植株的净光合速率逐渐回升,并最终恢复。（2）不同的光合参数对间歇性臭氧暴露敏感性不同,其中最大净光合速率最为敏感。在低AOT40下最大净光合速率显著降低,且恢复时间更长。（3） O₃二次暴露后,净光合速率降低幅度较低,且恢复更快,说明间歇性O₃暴露可能会提高大豆的耐受阈值。（4）当AOT40低于5.35μL L^-1 h^-1时,对大豆产量无显著影响,说明间歇性臭氧暴露条件下,大豆减产阈值更高。     

脑发育不同阶段慢性铅暴露对大鼠在体海马齿状回LTP的影响

目的和方法：探讨脑发育不同阶段慢性铅暴露对在体海马ＬＴＰ的影响。应用细胞外微电极记录单脉冲刺激穿通路纤维在海马齿状回诱发的群体锋电位（ＰＳ）,观察母体期、断乳后及出生前后持续性慢性铅暴露大鼠于高频刺激（ＨＦＳ）前后的ＰＳ幅值变化,并与对照组相比较。结果：ＨＦＳ前,基线记录的各铅暴露组ＰＳ平均幅值及峰潜伏期与对照组无显著差异;ＨＦＳ后,各铅暴露组ＬＴＰ发生率虽与对照组无显著差异,但铅暴露组的ＬＴＰ增幅减小,并出现了短时增强转为抑制及ＬＴＤ型反应。统计显示各铅暴露组ＨＦＳ后ＰＳ振幅的平均增强率显著低于对照组：对照组平均增强至基线值的１３８．２％,母体期铅暴露组为基线值的１０８．８％,断乳后铅暴露组为基线值的１０７．８％,持续铅暴露组为基线值的１０４．４％。结论：脑发育任一阶段的慢性铅暴露均可损害海马ＬＴＰ的在体诱导和维持,且以维持过程受损为主;与发育成熟海马相比,未成熟期海马对铅的神经毒性更为敏感,突触可塑性更易受损。     

臭氧急性暴露对大鼠血管的损伤及其机制

目的:探索不同浓度臭氧(O3)急性暴露对雄性Wistar大鼠血管的损伤效应和可能的机制。方法:120只雄性Wistar大鼠随机分为6组,每组20只;实验动物置于气体染毒柜中,对照组暴露于过滤后空气,处理组分别暴露于浓度为0.12ppm,0.5ppm,1.0ppm,2.0ppm和4.0ppm的臭氧,持续暴露4h。利用PC-lab医学生理信号采集系统获得动脉血压数据;血流变指标和血生化指标由天津迪安诊断实验室检测;血清中内皮素(ET-1)、同型半胱氨酸(HCY)、血管性血友病因子(vWF)、8-羟基脱氧鸟苷(8-OhdG)、白介素(IL-6)和肿瘤坏死因子α(TNF-α)采用酶联免疫(ELISA)微孔板法检测;氧化应激指标超氧化物歧化酶(SOD)活力和丙二醛(MDA)分别采用黄嘌呤氧化酶法、硫代巴比妥酸(TBA)法测定,还原型谷胱甘肽(GSH)和一氧化氮(NO)采用微孔板比色法;取胸主动脉组织制备石蜡切片,经HE染色后观察血管结构改变。结果:0.12ppm臭氧急性暴露可导致动脉收缩血压(SBP)显著升高;不同浓度臭氧暴露均可导致血浆粘度显著升高,1.0ppm臭氧暴露组血沉(ESR)方程K值显著升高,全血高切相对指数和还原粘度均在臭氧浓度为0.5ppm和4.0ppm时显著降低,而红细胞变形指数在臭氧浓度为0.12ppm、0.5ppm、1.0ppm和2.0ppm时显著升高;急性臭氧暴露可导致总胆固醇含量降低,高密度脂蛋白胆固醇(HDL-C)在0.12ppm臭氧暴露组显著降低;当臭氧浓度高于1.0ppm时还可导致机体出现炎症反应(TNF-α升高)和氧化应激反应(MDA升高、GSH降低);臭氧急性暴露可导致血液中ET-1含量升高,在4.0ppm浓度组具有显著性差异,而HCY水平呈现先降低后升高的趋势,在1.0ppm浓度组达到最高值,胸主动脉未见明显的病理改变。结论:臭氧急性暴露可影响大鼠的动脉血压、血流变及胆固醇代谢,可能的机制是臭氧暴露导致炎症反应和氧化应激反应,引起血管内皮功能损伤,并且随着臭氧暴露浓度升高血管内皮细胞功能损伤越显著。     

lncRNA与慢性肾病关系的研究进展

近年来,慢性肾病(chronic kidney disease,CKD)的发病率和死亡率逐年升高,越来越引起人们的重视。其发病机制复杂,病情往往持续性进展,最终发展为终末期肾功能衰竭。lncRNA是长链非编码RNA,显示出高度组织特异性,在最近一些研究中暗示了其在疾病中的重要性,lncRNA可能成为人类疾病未来治疗靶点。本文综述lncRNA在慢性肾病中的研究进展,为科学研究和临床应用提供参考。     

两种家猪心脏组织基因表达谱的分析

选取丹麦成年长白母猪的心脏、肌肉、主动脉、左心室和中国成年二花脸母猪心脏共5种组织,建立相应的cDNA文库,在此基础上测定了35180条表达序列标签。通过比较分析长白猪4种组织的基因表达,找出了长白猪心脏组织中3个显著高表达的功能群和47个显著高表达的基因。长白猪心脏组织在分子伴侣活动、马达活动和生理过程中表达的基因较多,47个显著高表达的基因都是和运动、运动调节、能量及保护有关的基因。两种家猪心脏组织表达差异显著的基因有74个,并且绝大部分是在长白猪心脏中表达更高,显示出长白猪心脏组织生理活动的高效与旺盛。     

Effects of sub-chronic aluminum chloride exposure on rat ovaries

Aims

This experiment investigated the effects of sub-chronic aluminum chloride (AlCl₃) exposure on rat ovaries.

Main methods

Eighty female Wistar (5 weeks old) rats, weighed 110–120 g, were randomly divided into four treatment groups: control group (CG), low-dose group (LG, 64 mg/kg BW AlCl₃), mid-dose group (MG, 128 mg/kg BW AlCl₃) and high-dose group (HG, 256 mg/kg BW AlCl₃). The AlCl₃ was administered in drinking water for 120 days. The ovarian ultrastructure was observed. The activities of acid phosphatase (ACP), alkaline phosphatase (ALP), succinate dehydrogenase (SDH), Na⁺–K⁺-ATPase, Mg^2 +-ATPase and Ca^2 +-ATPase, the contents of Fe, Cu and Zn, and the protein expression of follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) in the ovary were determined.

Key findings

The results showed that the structure of the ovary was disrupted, the activities of ALP, ACP, SDH, Na⁺–K⁺-ATPase, Mg^2 +-ATPase and Ca^2 +-ATPase, the contents of Zn, Fe and the protein expression of FSHR and LHR were lowered, and the content of Cu was increased in AlCl₃-treated rats than those in control.

Significance

The results indicate that sub-chronic AlCl₃ exposure caused the damage of the ovarian structure, the disturbed metabolism of Fe, Zn and Cu and the decreased activities of Na⁺–K⁺-ATPase, Mg^2 +-ATPase and Ca^2 +-ATPase in the ovary, which could result in suppressed energy supply in the ovary. A combination of suppression of energy supply and reduction of expression of FSHR and LHR could inhibit ovulation and corpus luteum development, leading to infertility in female rats.     

LncRNA‐mRNA expression profiles and functional networks in osteoclast differentiation

Human osteoclasts are differentiated from CD14⁺ monocytes and are responsible for bone resorption. Long non‐coding RNAs (lncRNAs) have been proved to be significantly involved in multiple biologic processes, especially in cell differentiation. However, the effect of lncRNAs in osteoclast differentiation is less appreciated. In our study, RNA sequencing (RNA‐seq) was used to identify the expression profiles of lncRNAs and mRNAs in osteoclast differentiation. The results demonstrated that expressions of 1117 lncRNAs and 296 mRNAs were significantly altered after osteoclast differentiation. qRT‐PCR assays were performed to confirm the expression profiles, and the results were almost consistent with the RNA‐seq data. GO and KEGG analyses were used to predict the functions of these differentially expressed mRNA and lncRNAs. The Path‐net analysis demonstrated that MAPK pathway, PI3K‐AKT pathway and NF‐kappa B pathway played important roles in osteoclast differentiation. Co‐expression networks and competing endogenous RNA networks indicated that ENSG00000257764.2‐miR‐106a‐5p‐TIMP2 may play a central role in osteoclast differentiation. Our study provides a foundation to further understand the role and underlying mechanism of lncRNAs in osteoclast differentiation, in which many of them could be potential targets for bone metabolic disease.     

大气臭氧浓度升高对银杏叶片活性氧代谢及相关基因表达的影响

采用开顶式气室熏蒸法,设置自然条件下臭氧(O₃)浓度(对照,约40 nmol·mol^-1)、80、160及200 nmol·mol^-14个臭氧浓度,观测了不同浓度臭氧条件下银杏叶片可见伤害、活性氧生成量、抗氧化酶活性及相关基因表达变化情况,分析大气臭氧浓度升高对植物活性氧代谢的影响.结果表明: 160和200 nmol·mol^-1 O₃熏蒸明显伤害银杏叶片,80 nmol·mol^-1与对照无差异,无可见伤害.O₃处理20 d后,160和200 nmol·mol^-1条件下银杏叶片的超氧自由基(O₂^-·)产生速率显著高于80 nmol·mol^-1和对照,而80 nmol·mol^-1与对照无差异;O₃处理40 d后,160和200 nmol·mol^-1熏蒸下叶片过氧化氢(H₂O₂)含量显著高于80 nmol·mol^-1和对照,而过氧化氢酶(CAT)活性显著高于80 nmol·mol^-1和对照,各臭氧处理抗坏血酸过氧化物酶(APX)活性均低于对照.熏蒸40 d后,CAT、APX基因的转录表达持续加强;防御素(GbD)的表达强度则随着臭氧浓度的增加及熏蒸时间的延长而呈显著加强.高浓度臭氧胁迫可使银杏叶片活性氧生成量增加、抗氧化酶活性下降、相关基因表达水平上调,有明显可见叶片伤害.     

Integrative analysis of genome‐wide lncRNA and mRNA expression in newly synthesized Brassica hexaploids

Polyploidization, as a significant evolution force, has been considered to facilitate plant diversity. The expression levels of lncRNAs and how they control the expression of protein‐coding genes in allopolyploids remain largely unknown. In this study, lncRNA expression profiles were compared between Brassica hexaploid and its parents using a high‐throughput sequencing approach. A total of 2,725, 1,672, and 2,810 lncRNAs were discovered in Brassica rapa, Brassica carinata, and Brassica hexaploid, respectively. It was also discovered that 725 lncRNAs were differentially expressed between Brassica hexaploid and its parents, and 379 lncRNAs were nonadditively expressed in this hexaploid. LncRNAs have multiple expression patterns between Brassica hexaploid and its parents and show paternal parent‐biased expression. These lncRNAs were found to implement regulatory functions directly in the long‐chain form, and acted as precursors or targets of miRNAs. According to the prediction of the targets of differentially expressed lncRNAs, 109 lncRNAs were annotated, and their target genes were involved in the metabolic process, pigmentation, reproduction, exposure to stimulus, biological regulation, and so on. Compared with the paternal parent, differentially expressed lncRNAs between Brassica hexaploid and its maternal parent participated in more regulation pathways. Additionally, 61 lncRNAs were identified as putative targets of known miRNAs, and 15 other lncRNAs worked as precursors of miRNAs. Some conservative motifs of lncRNAs from different groups were detected, which indicated that these motifs could be responsible for their regulatory roles. Our findings may provide a reference for the further study of the function and action mechanisms of lncRNAs during plant evolution.     

The role of low-level ozone exposure and mycorrhizas in chemical quality and insect herbivore performance on Scots pine seedlings

The effects of low‐level ozone exposure and suppression of natural mycorrhizas on the above‐ground chemical quality of Scots pine (Pinus sylvestris L.) needles and insect herbivore performance were studied in a two‐year field experiment. Seedlings were fumigated with the ozone doses 1.5–1.7 times the ambient, and natural mycorrhizal infection level was about 35% reduced in roots with fungicide propiconazole. On ozone‐exposed seedlings the mean relative growth rate (MRGR) of Lygus rugulipennis Popp. nymphs was lower than on ambient ozone seedlings, but Gilpinia pallida Klug sawfly larvae grew better on elevated ozone seedlings than on ambient ozone seedlings. MRGR of Schizolachnus pineti Fabr. and Cinara pinea L. aphid nymphs or Neodiprion sertifer Geoffr. sawfly larvae or the oviposition of L. rugulipennis and N. sertifer were not affected by ozone exposure. Although ozone exposure did not affect total phenolics, total terpene, total or individual resin acid, total free amino acid, nutrient or sugar concentrations in needles, MRGR of L. rugulipennis positively correlated with total terpenes and MRGR of G. pallida positively with total amino acids. In addition, ozone exposure increased serine and proline concentration and marginally also starch concentration in needles. When mycorrhizas were reduced with fungicide, only MRGR of L. rugulipennis nymphs increased, but performance of other insect herbivores studied was not changed. However, number of L. rugulipennis eggs correlated positively with mycorrhizal infection level and also with total sugars. Reduction of mycorrhizas did not strongly affect the concentrations of analysed compounds in needles, because only phosphorus and potassium and some individual resin acids were reduced by fungicide treatment. These results suggest that low‐level ozone exposure and moderately declined mycorrhizal infection do not drastically affect either the above‐ground chemical quality of Scots pine seedlings or performance of studied insect herbivores.     

Long-term effects of dietary isoflavones on uterine gene expression profiles

Isoflavones (ISOs) are bioactive food ingredients of the traditional East Asian diet and currently discussed as alternatives to classical hormone replacement therapies and for reducing the prevalence of hormone-dependent cancers. Although there are many studies on ISOs, not much is known about their long-term effects.Therefore, we performed an animal experiment analyzing the effects of three different diets: a phytoestrogen-free diet, a diet supplemented with genistein (700 μg/g diet) and an ISO-high diet (232 μg daidzein and 240 μg genistein/g) at two distinct time points, juvenile (21 days) and adult (97 days). Exposure started prior to mating of the parents and throughout the life of the offspring.We observed a stronger increase of uterine wet weights in juvenile offspring with genistein exposure (1018 ± 350 mg/kg BW) than with ISO-high diet (497 ± 133 mg/kg BW). Whereas the expression of proliferation related genes (PCNA; Ki67; IGF-1; IGF-1R), analyzed by real-time-qPCR and Western blot, were significantly down-regulated in juvenile animals exposed to genistein. Additionally, genistein exposure led to estrogenic responses, observed upon increase of complement C3 and decrease of estrogen receptors gene expressions, while the exposure to ISO-high diet did not show these effects.In conclusion, both the time point on which phytoestrogen exposure starts together with the composition of the ingested phytoestrogen containing diet are of great importance for the biological response of the offspring.     

基因芯片技术与基因表达谱研究

基因芯片技术是近年来出现的分子生物学与微电子技术相结合的最新DNA分析检测技术,该技术将成为信息科学与生命科学之间的联系纽带,为后基因组时代基因功能的分析提供一种最重要的技术手段,目前基因芯片技术已在基因表达谱等研究中得到广泛应用。