杂交粳稻花优14及其亲本孕穗期剑叶的基因表达谱芯片分析

本文以孕穗期的杂交粳稻花优14及其母本申9A和父本繁14的剑叶为材料,利用Affymetrix水稻基因组芯片检测了3个样品中的基因表达谱,并用生物信息学方法对差异表达基因进行了分析。结果表明：与其亲本相比,杂交粳稻花优14中共有2057个基因的表达水平出现了2倍(变化倍数≥2或≤0.5)以上的变化。通过对这些差异表达基因进行GO(Gene Ontology)功能分类,发现差异表达基因在光合系统Ⅰ、叶绿体膜和叶绿体被膜等与叶绿体相关的细胞组分中显著富集;同时差异表达基因还在叶绿素合成、叶绿素代谢和类胡萝卜素合成等生物学过程中富集。光合作用效率的改变可能和花优14杂种优势的形成相关。与已报道结果不同,本文在代谢途径分析结果中并没有发现花优14中差异表达基因在碳固定和光合作用等途径中显著的富集,但是发现差异表达基因在光合作用-天线蛋白以及淀粉和蔗糖的代谢途径中显著富集。该结果表明,在不同的杂交组合中,参与杂种优势形成的基因或者代谢途径可能是不同的。     

鸡miR-9不同组织表达差异及其功能预测分析

miRNA在动物生长发育过程中有重要作用. 本文采用定制茎环反转录引物,利用实时荧光定量PCR技术构建miR-9在鸡2个阶段11个组织中的表达谱,同时用TargetScan5.1与PicTar两种计算方法对其进行靶基因预测,交集的基因集合分别进行GO（gene ontology）富集分析和生物通路富集分析. 结果表明,采用实时定量PCR检测的miR-9在鸡下丘脑中的表达量和高通量测序结果一致;采用实时定量PCR在对不同组织定量结果表明,miR-9在0日龄鸡的肾脏、下丘脑、腿肌和大脑中高丰度表达,在成年鸡表达量较高为大脑、腿肌、心脏、小脑和下丘脑.在同一组织的0日龄和成年鸡中表达呈现时序性,除肝脏的表达量差异不显著,其他10个组织miR-9表达量差异显著（P<0.05）.预测到交集靶基因有160个.涉及到多个KEGG通路和GO富集中.GO分类结果显示,这些基因分布于63个群中,其中基因超过45个基因群集有26个群,与代谢有关的群有11个. 其它与发育、调控等过程有关. 在KEGG通路分析中,显著的通路有细胞骨架调控、细胞增殖和分化有关的通路（P<0.01）等5个通路. 表明miR 9基因的表达有组织和时序特异性,靶基因参与细胞代谢、生长发育和调控.这些结果为进一步验证miR 9基因在在脑中调控生长发育过程中的作用奠定了基础.     

大肠杆菌应激诱导基因表达谱芯片分析

应激是影响微生物发酵生产的重要因素。为挖掘细菌耐热和化合物胁迫等调控相关基因,揭示微生物应激反应调控的分子机制,采用Affymetrix全基因组表达谱芯片,对42℃热激和0.1 mol/L CaCl2处理的大肠杆菌E. coli基因谱进行分析。结果表明,应激处理菌体中共有583个差异表达基因,包括374个表达量上调和209个表达量下调差异基因;GO功能分类将差异表达基因分为34类,大部分差异基因的分子功能覆盖结合、催化和转运等生物学活性;差异基因KEGG分析显示富集在28个通路,以ABC转运蛋白、鞭毛组装、双组分系统、尿素循环和氨基酸新陈代谢以及Ⅲ型分泌系统等通路为主要代表;应激胁迫的E. coli主要通过噬菌体休克蛋白操纵子(psp)基因进行逆境适应性转录调节,同时伴随着酶和氨基酸的合成基因以及细胞氧化还原态平衡基因的调控变化。     

虹鳟肝组织新转录本分析及基因结构优化

目的虹鳟热应激下肝RNA-seq数据中新转录本的分析及已注释基因结构优化。方法以虹鳟肝为材料提取总RNA,构建cDNA文库,并利用Illumina双端测序Hiseq 2500平台进行测序。运用Cufflinks软件对测序数据进行组装,将其与虹鳟参考基因组进行序列比对。结果发掘新转录本6555个,其中30个新转录本在热应激前后差异表达(P<0.05)。与GO数据库比对对新转录本进行功能注释,获得3097个新转录本的注释。与KEGG数据库比对,共有3617个新转录本注释到284条代谢通路中。对19 424个已注释基因的结构进行优化,延伸了14 719个基因的5′端和14 796个基因的3′端。结论通过对发掘的6555个新转录本分析,并对19 424个已注释基因结构优化,为虹鳟基因组注释信息的完善提供了有力的借鉴,并为进一步了解虹鳟热应激的机制提供更有力的理论基础。     

利用基因芯片技术筛选秦川牛公牛与阉牛肌肉组织差异表达基因

为了筛选秦川牛公牛和阉牛肌肉组织差异表达的基因,探讨二者肉质差异的分子生物学机理,文章利用Affymetrix公司生产的牛基因组芯片技术,分别检测了3头36月龄秦川牛公牛与阉牛背最长肌肌肉组织的mRNA表达水平变化;运用Significance Analysis of Microarrays(SAM)法对秦川牛公牛和阉牛基因表达谱进行了差异分析;并通过分子注释系统平台(MAS2.0)对差异表达基因进行了功能富集类分析和调控通路分析;最后应用实时定量PCR技术对部分差异表达基因进行了验证。基因表达谱分析结果显示,在36月龄秦川牛的肌肉组织中,共检测到约11000个探针,代表大约10000个基因。共筛选出差异表达基因143条,主要涉及胶原纤维的组织和合成、细胞粘附、细胞生长调控、泛素介导的蛋白分解代谢和横纹肌收缩等生物学过程;在分子注释系统数据库中注释到的显著调控通路为细胞外基质受体反应、细胞通讯、焦点粘连和紧密连接等;所验证的差异表达基因的荧光定量PCR结果与芯片结果基本一致。结合已有的文献报道,文章初步认为ECM受体反应、细胞通讯、焦点粘连、紧密接头等调控通路及COL3A1、COL1A1、COL1A2、SPP1、FBN1、MMP2、ECM1、MYH3、MYH8、S100A4、ASPN、CFD等基因可能是参与调控秦川牛阉割前后肉质性状差异的重要调控通路和基因。此外,还筛选出一些尚未在GenBank上登陆的序列,推测可能是未知的新基因,它们在牛肉质代谢过程中的作用还需进一步的研究证明。     

氧化鱼油对草鱼肠道黏膜抗氧化应激通路基因表达水平的影响

为了探讨氧化鱼油对草鱼肠道黏膜损伤后, 参与抗氧化应激的基因通路及其通路基因表达活性的变化,以草鱼为试验对象, 灌喂氧化鱼油7d后, 采集肠道黏膜组织并提取总RNA, 采用RNA-seq方法, 进行了氧化鱼油组和正常鱼油组草鱼肠道黏膜基因注释、IPA基因通路分析和基因表达活性差异分析。结果显示, 组织切片观察发现氧化鱼油导致草鱼肠道黏膜出现严重的损伤; 肠道黏膜中具有较为完整的Keap1-Nrf2-ARE基因调控通路。肠道黏膜在受到氧化鱼油的氧化损伤作用后, 激活了细胞的抗氧化损伤保护机制, 使NRF2介导的氧化应激反应通路基因差异表达显著性地上调, 并导致了下游的GSH/GSTs通路基因差异表达显著性上调, 促进了GSH的生物合成和GSTs的抗氧化作用; 导致Keap1-Nrf2-ARE信号通路下游的热休克蛋白和泛素-蛋白酶体通路基因差异表达显著性上调, 清除受损伤蛋白质, 保护细胞结构完整性。研究表明, 上述三类抗氧化应激通路构成了对肠道黏膜损伤细胞、损伤蛋白质的降解系统和清除系统, 显示其对肠道黏膜组织和黏膜细胞的保护、修复发挥了重要的作用。     

Keap1/Nrf2/ARE通路相关基因在热诱导的氧化应激小鼠肝脏中的表达

为了研究热应激对小鼠肝脏抗氧化功能及Keap1 (kelch-like ECH-associated protein- 1)/Nrf2（NF-E2-related factor 2）/ARE (antioxidant response element)通路相关基因表达的影响,选用30只8周龄雄性小鼠随机分成6组,每 d连续42 ℃热处理2 h,分别在热处理0 d(对照组)、1 d、2 d、4 d、8 d和12 d时观察肝脏组织形态学和免疫组织化学分析,另取一部分肝脏组织保存于-80 ℃用于后续荧光定量PCR实验,检测肝脏抗氧化指标及Keap1/Nrf2/ARE通路相关基因的表达．结果显示：小鼠的体表温度和直肠温度在热处理后都极显著高于热处理前．组织形态学观察发现,热处理导致小鼠肝脏组织充血和肝细胞水肿．小鼠肝脏氧化应激指标 MDA (malondialdehyde)含量在热处理第2 d较对照组显著升高,GSH (glutathione)含量、GSH-PX (glutathione peroxidase)活力和总SOD (superoxide dismutase)活力在第4 d和12 d都有升高．免疫组织化学发现,与对照组和第12 d组相比,Nrf2蛋白在第1 d,2 d,4 d,8 d表达明显,其中Nrf2蛋白在第4 d表达最为显著. 荧光定量RT PCR结果表明,与对照组比较Keap1基因的表达量从热处理第1 d开始显著降低,Nrf2基因的表达量在第4 d和12 d显著升高,HO-1 (Heme oxygenase-1)基因的表达量在第1 d显著升高,NQO1 (Quinone oxidoreductase)和GCLC (Glutamate cysteine ligase catalytic)基因的表达量在第1 d和4 d显著升高．上述结果表明,热应激引起了小鼠肝脏氧化损伤, Keap1/Nrf2/ARE通路可能参与了肝脏自身缓解热应激的过程.     

感染球孢白僵菌的家蚕免疫应答差异表达基因分析

【目的】筛选家蚕Bombyx mori应对白僵菌Beauveria bassiana侵染的应答基因, 以进一步研究家蚕抵御真菌侵染的分子机制。【方法】采用新一代Solexa高通量测序技术对感染白僵菌及未感染白僵菌的对照组家蚕进行了测序分析, 筛选差异表达基因; 结合生物信息学工具分析差异表达基因的功能注释、 分类及涉及的信号通路等; 应用荧光定量PCR技术验证10个基因的差异表达。【结果】通过测序和生物信息学分析共获得377个差异表达基因, 其中表达上调基因236个, 下调基因141个; KEGG通路分析表明, 各通路中既有表达上调的基因, 也有下调基因; 12个上调基因、 26个下调基因参与3个显著性富集的KEGG通路, 即核糖体、 氨酰tRNA生物合成和剪接体通路。定量PCR与测序结果显示, 溶菌酶、 热激蛋白、 谷胱甘肽S-转移酶、 肽聚糖识别蛋白等与免疫应激相关的蛋白基因均呈现表达上调。【结论】本研究筛选获得的差异表达基因, 特别是上调表达的基因可能与家蚕应对白僵菌侵染的应答机制有关, 其中与免疫应激相关的蛋白基因如溶菌酶、 热激蛋白、 谷胱甘肽S 转移酶、 肽聚糖识别蛋白基因等可能直接参与了家蚕对白僵菌的免疫识别和防御, 研究结果为从分子水平阐明家蚕抵御真菌侵染的防御机制和白僵菌对家蚕的致病机理提供新的依据。     

Affymetrix基因表达谱芯片在新疆维吾尔族与汉族胰腺癌组织样本间差异表达基因筛选中的运用

目的:运用基因表达谱芯片筛选并分析新疆维吾尔族与汉族胰腺癌组织样本间的差异表达基因。方法:收集我院2014年1月至2016年6月间行手术切除的维吾尔族与汉族胰腺导管细胞癌组织并提取总RNA,选取经Nanodrop 2000与Agilent 2100仪器质检合格的样本总RNA采用Affymetrix基因表达谱芯片筛选出差异表达基因并绘制统计图,运用基因本体(GO)分析及信号通路(Pathway)分析对这些差异表达基因的生物信息进行汇总分析。结果:通过基因表达谱芯片分析,新疆维吾尔族与汉族胰腺癌组织样本间共检测到1063个基因存在差异表达,在维吾尔族胰腺癌标本中显著上调表达的基因共281个,差异表达倍数最高的为IGLV1-44基因(差异倍数:9.99)下调表达的基因共782个,差异表达倍数最高的为CPB1基因(差异倍数:33.76);在Gene Ontology数据库中共检索到815个上述差异表达基因具有明确的GO分类,差异表达倍数最高的为CPB1基因(差异倍数:33.76);Pathway分析中共检测到30条信号通路包含有上述差异表达基因,共涉及196个基因,其中以FAK信号通路差异表达基因富集程度最高,差异表达倍数最高的基因为COL11A1基因(差异倍数:5.02)。结论:基因表达谱芯片分析结果显示,在新疆维吾尔族与汉族胰腺癌组织样本间存在大量的差异表达基因,这些基因与胰腺癌的增殖分化、侵袭转移及多药耐药等特性密切相关,且参与了多条生物体内重要信号转导通路的调控。     

6天龄与10天龄大鼠的睾丸组织的基因差异表达

通过基因芯片技术,利用Roche-NimbleGen公司制作的大鼠12×135K全基因组表达谱芯片,对日龄为6d和10d的大鼠睾丸组织进行全基因组表达差异分析。结果显示:具有2倍以上的差异表达基因有4298个,其中表达上调的基因共1878个,表达下调的基因共2420个。这些差异表达的基因中有3154个基因具有基因本体注释,参与了154个生物学通路。进一步分析表明具有8倍以上差异表达的基因有13个,这些基因参与了生物学过程、细胞组分和分子功能等基因本体分类,进一步选择3个差异表达的基因,LOC686076、Cxcl6和Trib3,做了实时定量RT-PCR检测。其结果趋势与芯片数据一致。因此,我们初步认为精原干细胞的发生与增殖在大鼠早期的发育过程中已经有大量的基因参与,是一个多基因协调表达的过程。     

Protective role of 5‐azacytidine on myocardial infarction is associated with modulation of macrophage phenotype and inhibition of fibrosis

We examined whether a shift in macrophage phenotype could be therapeutic for myocardial infarction (MI). The mouse macrophage cell line RAW264.7 was stimulated with peptidoglycan (PGN), with or without 5-azacytidine (5AZ) treatment. MI was induced by ligation of the left anterior descending coronary artery in rats, and the rats were divided into two groups; a saline-injection group and a 5AZ-injection group (2.5 mg/kg/day, intraperitoneal injection). LV function was evaluated and immunohistochemical analyses were performed 2 weeks after MI. Cardiac fibrosis was induced by angiotensin II (AngII) infusion with or without 5AZ (5 mg/kg/day) in mice. Nitric oxide was produced by PGN, which was reduced by 77.87% after 5AZ treatment. Both induction of inducible nitric oxide synthase (iNOS) and iNOS promoter activity by PGN were inhibited by 5AZ. Ejection fraction (59.00 ± 8.03% versus 42.52 ± 2.58%), contractility (LV dP/dt-max, 8299.76 ± 411.56 mmHg versus 6610.36 ± 282.37 mmHg) and relaxation indices (LV dP/dt-min, −4661.37 ± 210.73 mmHg versus −4219.50 ± 162.98 mmHg) were improved after 5AZ administration. Cardiac fibrosis in the MI+5AZ was 8.14 ± 1.00%, compared with 14.93 ± 2.98% in the MI group (P < 0.05). Arginase-1(+)CD68(+) macrophages with anti-inflammatory phenotype were predominant in the infarct border zone of the MI+5AZ group, in comparison with the MI group. AngII-induced cardiac fibrosis was also attenuated after 5AZ administration. In cardiac fibroblasts, pro-fibrotic mediators and cell proliferation were increased by AngII, and these increases were attenuated after 5AZ treatment. 5AZ exerts its cardiac protective role through modulation of macrophages and cardiac fibroblasts.     

Metabolic Consequences of High-Fat Diet Are Attenuated by Suppression of HIF-1��

Obesity is associated with tissue hypoxia and the up-regulation of hypoxia inducible factor 1 alpha (HIF-1α). Prior studies in transgenic mice have shown that HIF-1α plays a role in the metabolic dysfunction associated with obesity. Therefore, we hypothesized that, after the development of diet-induced obesity (DIO), metabolic function could be improved by administration of HIF-1α antisense oligonucleotides (ASO). DIO mice were treated with HIF-1α ASO or with control ASO for 8 weeks and compared with an untreated group. We found that HIF-1α ASO markedly suppressed Hif-1α gene expression in adipose tissue and the liver. HIF-1α ASO administration induced weight loss. Final body weight was 41.6±1.4 g in the HIF-1α ASO group vs 46.7±0.9 g in the control ASO group and 47.9±0.8 g in untreated mice (p<0.001). HIF-1α ASO increased energy expenditure (13.3±0.6 vs 12±0.1 and 11.9±0.4 kcal/kg/hr, respectively, p<0.001) and decreased the respiratory exchange ratio (0.71±0.01 vs 0.75±0.01 and 0.76±0.01, respectively, p<0.001), which suggested switching metabolism to fat oxidation. In contrast, HIF-1a ASO had no effect on food intake or activity. HIF-1α ASO treatment decreased fasting blood glucose (195.5±8.4 mg/dl vs 239±7.8 mg/dl in the control ASO group and 222±8.2 mg/dl in untreated mice, p<0.01), plasma insulin, hepatic glucose output, and liver fat content. These findings demonstrate that the metabolic consequences of DIO are attenuated by HIF-1α ASO treatment.     

Status of Systemic Oxidative Stresses in Patients with Primary Open-Angle Glaucoma and Pseudoexfoliation Syndrome

Background

The involvement of local and systemic oxidative stress in intraocular pressure (IOP) elevation and optic nerve damage has been hypothesized in the pathogenesis of glaucoma. To test this, we measured the systemic levels of prooxidants and antioxidants by analyzing the blood biochemistry in patients with glaucoma.

Methods

Peripheral blood samples were collected from Japanese patients with primary open-angle glaucoma (PG) (n = 206), exfoliation syndrome (EX) (n = 199), and controls (n = 126). Plasma levels of lipid peroxides, ferric-reducing activity, and thiol antioxidant activity were measured by diacron reactive oxygen metabolites (dROM), biological antioxidant potential (BAP), and sulfhydryl (SH) tests, respectively, using a free radical analyzer.

Results

In the PG, EX, and control groups, the mean ± standard deviation values were 355±63, 357±69, and 348±56 (U. Carr), respectively, for dROM; 1,951±282, 1,969±252, and 2,033±252 (µmol/L), respectively, for BAP (µmol/L); and 614±98, 584±91, and 617±99 (µmol/L), respectively, for SH. The differences in the BAP values were significant between the PG and control groups (p = 0.0062), for SH between the EX and control groups (p = 0.0017), and for SH between the PG and EX groups (p = 0.0026). After adjustment for differences in age and sex among groups using multiple regression analysis, lower BAP values were correlated significantly with PG (p = 0.0155) and EX (p = 0.0049). Higher dROM values with and without glaucoma were correlated with female gender, and lower SH values with older age. There were no significant differences between the higher (≥21 mmHg) and lower (<21 mmHg) baseline IOPs in the PG group or between the presence or absence of glaucoma in the EX group.

Conclusions

Lower systemic antioxidant capacity that measured by ferric-reducing activity is involved in the pathogenesis of PG and EX.     

In vivo comparison of the bone regeneration capability of human bone marrow concentrates vs. platelet-rich plasma

Background

Bone marrow aspirate concentrate (BMAC) including high densities of stem cells and progenitor cells may possess a stronger bone regenerative capability compared with Platelet-rich plasma (PRP), which contains enriched growth factors. The objective of this study was to evaluate the effects of human BMAC and PRP in combination with β-tricalcium phosphate (β-TCP) on promoting initial bone augmentation in an immunodeficient mouse model.

Methodology/Principal Findings

BMAC and PRP were concentrated with an automated blood separator from the bone marrow and peripheral blood aspirates. β-TCP particles were employed as a scaffold to carry cells. After cell counting and FACS characterization, three groups of nude mice (BMAC+TCP, PRP+TCP, and a TCP control) were implanted with graft materials for onlay placement on the cranium. Samples were harvested after 4 weeks, and serial sections were prepared. We observed the new bone on light microscopy and performed histomorphometric analysis. After centrifugation, the concentrations of nucleated cells and platelets in BMAC were increased by factors of 2.8±0.8 and 5.3±2.4, respectively, whereas leucocytes and platelets in PRP were increased by factors of 4.1±1.8 and 4.4±1.9, respectively. The concentrations of CD34-, CD271-, CD90-, CD105-, and CD146-positive cells were markedly increased in both BMAC and PRP. The percentage of new bone in the BMAC group (7.6±3.9%) and the PRP group (7.2±3.8%) were significantly higher than that of TCP group (2.7±1.4%). Significantly more bone cells in the new bone occurred in sites transplanted with BMAC (552±257) and PRP (491±211) compared to TCP alone (187±94). But the difference between the treatment groups was not significant.

Conclusions/Significance

Both human BMACs and PRP may provide therapeutic benefits in bone tissue engineering applications. These fractions possess a similar ability to enhance early-phase bone regeneration.     

Associations between body mass and the outcome of surgery for scoliosis in Chinese adults

Background

In this study we intended to prove that being overweight has an unfavorable impact on the surgical treatment outcome of adult idiopathic scoliosis (AdIS).

Methods

This is a retrospective study on the surgical treatment of seventy-one more than 30 years old (58 females and 13 males; mean age 42.9±12.2) idiopathic scoliotic patients with a minimum follow up of at least 2 years. The patients were divided into an overweight group (BMI≥23) and a non-overweight group (BMI<23). Preoperative, postoperative first erect and final follow-up radiographic measures, perioperative data, the Oswestry disability index (ODI), and the visual analog scale (VAS) were reviewed and compared.

Findings

In the overweight group, no significant differences in radiographic measures, perioperative data, preoperative comorbidities, or postoperative complications, except for the more frequent concomitance of preoperative thoracic kyphosis 37.9±7.7 vs. 26.5±11.8 (P = 0.000) and thoracolumbar kyphosis 14.9±10.1 overweighted group vs. 6.5±9.9 non-overweighted group respectively (P = 0.002) were found. A higher morbidity of hypertension 36.8% vs. 9.6% (P = 0.004) was also observed in the overweight group. Postoperative ODI and VAS improved significantly in both groups compared to pre-operative values. The postoperative ODI of the overweight group (19.6±12.4) was significantly higher than that of the non-overweight group (12.4±7.9) (P = 0.022).

Conclusions

Overweight adult idiopathic scoliotic patients had more frequent concomitance of preoperative thoracic kyphosis and thoracolumbar kyphosis and more serious postoperative pain. However, BMI did not affect the outcomes of surgical correction for coronal and sagittal scoliotic deformity and their postoperative complication rates were not significantly affected.     

Relative Bioavailability of Chlorothiazide from Mucoadhesive Compacts in Pigs

The relative bioavailability of chlorothiazide from mucoadhesive polymeric compacts is compared to commercial oral suspension in pigs. A single-dose randomized study was conducted in 12 healthy pigs that are 9–10 weeks old. After overnight fasting, pigs were divided into two groups of six animals. To the first group, a reference product containing 50 mg of chlorothiazide suspension, and in the second group, test product (mucoadhesive compacts) chlorothiazide (50 mg) was administered with 75 mL of water via gastric tubes. Blood samples were collected between 0 to 24 h using catheters inserted into the jugular vein. Plasma was separated by protein precipitation, and chlorothiazide concentrations were determined using a high-performance liquid chromatography method. The mean T_max and the C_max of chlorothiazide following the administration of oral suspension and mucoadhesive compacts were 0.58 ± 0.20 h and 682.97 ± 415.69 ng/mL and 2.17 ± 0.98 h and 99.42 ± 124.08 ng/mL, respectively. The K_el and T_1/2 of chlorothiazide were found to be 1.06 ± 0.28 h⁻¹ and 0.70 ± 0.21 h from suspension and 0.95 ± 1.11 h⁻¹ and 2.05 ± 1.90 h from the compacts, respectively. The T_max of mucoadhesive compacts were significantly longer (p < 0.05; 2.17 h) than the reference products (0.58 h), whereas the C_max of compacts were significantly lower (99 ng/mL) than the reference product (683 ng/mL; p < 0.05). The area under the curve (AUC) of compacts accounts only 50.15% (404.32 ± 449.93 ng h/mL) of the reference product’s AUC (806.27 ± 395.97 ng h/mL). The relative bioavailability of the compacts was lower than that of the suspension, and this may be due to the narrow window of absorption for chlorothiazide.Key words: bioavailability, chlorothiazide, mucoadhesive compacts, pigs     

Detection of Ammonia-Oxidizing Archaea in Fish Processing Effluent Treatment Plants

Ammonia oxidation is the rate limiting step in nitrification and thus have an important role in removal of ammonia in natural and engineered systems with participation of both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). However, their relative distribution and activity in fish processing effluent treatment plants (FPETPs) though significant, is hitherto unreported. Presence of AOA in sludge samples obtained from FPETPs was studied by amplification and sequencing of thaumarchaeal ammonia monooxygenase subunit A (AOA-amoA) gene. Different primer sets targeting 16S rRNA and AOA-amoA gene were used for the detection of AOA in FPETPs. Phylogenetic analysis of the gene revealed that the AOA was affiliated with thaumarchaeal group 1.1a lineage (marine cluster). Quantitative real time PCR of amoA gene was used to study the copy number of AOA and AOB in FPETPs. The AOA-amoA and AOB-amoA gene copy numbers of sludge samples ranged from 2.2 × 10⁶ to 4.2 × 10⁸ and 1.1 × 10⁷ to 8.5 × 10⁸ mg⁻¹ sludge respectively. Primer sets Arch-amoAF/Arch-amoAR and 340F/1000R were found to be useful for the sensitive detection of AOA-amoA and Archaeal 16S rRNA genes respectively in FPETPs. Their presence suggests the widespread occurrence and possible usefulness in removing ammonia from FPETPs which is in line with reports from other waste water treatment plants.

Electronic supplementary material

The online version of this article (doi:10.1007/s12088-014-0484-6) contains supplementary material, which is available to authorized users.     

肺动脉高压及其合并心脏内右向左分流患者的心肺运动试验特征性变化的临床研究*

目的: 明确肺动脉高压及合并心脏内右向左分流（R-L）患者的心肺运动试验（CPET）气体交换变化。方法: 本文通过回顾性分析阜外医院从2016-10至2017-08签署知情同意书后完成CPET的73例肺动脉高压病人CPET数据,采取双盲方式抽取四位医生作为判读者分别独立识别R-L后,结果分为四组：①分流阳性组（n=20）、②分流可疑组（n=9）、③无分流组（n=37）、④分流延迟开放组（n=6）。选择同期完成CPET正常人14例作为对照。结果: 分流阳性组在运动开始时分钟通气量、二氧化碳排出通气效率、氧气通气效率和呼气末氧分压相对于静息期的改变值骤升,分别为（7.36±2.72） L/min、（1.84±3.59）、（5.02±4.34）、（3.75±2.64） mmHg）,明显高于对照组的（（4.26±2.59） L/min、（2.22±2.08）、（-1.46±4.68）、（-3.96±2.82） mmHg）;而呼气末二氧化碳分压相对于静息期的改变值骤降（-1.63±1.66） mmHg,明显低于对照组的（2.22±2.08） mmHg（P均<0.01）。分流延迟开放组在运动后期呼吸商（RER）、二氧化碳排出通气效率、氧气通气效率和呼气末氧分压相对于静息期的改变值骤升,分别为（0.40±0.08）、（11.07±5.60）、（30.55±7.89）、（13.72±2.21） mmHg,明显高于对照组的（0.38±0.12）、（5.67±4.6）、（4.54±3.83）、（5.51±4.24） mmHg;而呼气末二氧化碳分压相对于静息期的改变值骤降（-6.82±1.96） mmHg,明显区别于对照组的（5.67±4.6） mmHg,在恢复期分流延迟开放组二氧化碳排出通气效率、氧气通气效率相对于峰值功率时的改变值（分别为-8.38±3.24、-13.14±6.47）,明显低于对照组（6.22±2.87、16.56±4.2）（P均<0.01）。结论: 肺动脉高压患者较正常人CPET的整体功能和通气效率指标降低;肺动脉高压合并右向左分流患者不仅在静息通气效率受限更剧;且特征性地运动初始时出现PETO₂明显上升、PETCO₂明显下降,RER跳升到1.0左右,VE/VCO₂ 不降反升与VO₂/VE不升反降, 常有SpO₂显著下降,还有VE更大幅度上升;延迟开放型上述特征性变化发生在运动接近峰值的1~3 min而非运动初始,且运动停止后迅速反向变回以示重新关闭。     

Expansion of the Multi-Link Frontier™ coronary bifurcation stent: micro-computed tomographic assessment in human autopsy and porcine heart samples

Background

Treatment of coronary bifurcation lesions remains challenging, beyond the introduction of drug eluting stents. Dedicated stent systems are available to improve the technical approach to the treatment of these lesions. However dedicated stent systems have so far not reduced the incidence of stent restenosis. The aim of this study was to assess the expansion of the Multi-Link (ML) Frontier™ stent in human and porcine coronary arteries to provide the cardiologist with useful in-vitro information for stent implantation and selection.

Methodology/Principal Findings

Nine ML Frontier™ stents were implanted in seven human autopsy heart samples with known coronary artery disease and five ML Frontier™ stents were implanted in five porcine hearts. Proximal, distal and side branch diameters (PD, DD, SBD, respectively), corresponding opening areas (PA, DA, SBA) and the mean stent length (L) were assessed by micro-computed tomography (micro-CT). PD and PA were significantly smaller in human autopsy heart samples than in porcine heart samples (3.54±0.47 mm vs. 4.04±0.22 mm, p = 0.048; 10.00±2.42 mm² vs. 12.84±1.38 mm², p = 0.034, respectively) and than those given by the manufacturer (3.54±0.47 mm vs. 4.03 mm, p = 0.014). L was smaller in human autopsy heart samples than in porcine heart samples, although data did not reach significance (16.66±1.30 mm vs. 17.30±0.51 mm, p = 0.32), and significantly smaller than that given by the manufacturer (16.66±1.30 mm vs. 18 mm, p = 0.015).

Conclusions/Significance

Micro-CT is a feasible tool for exact surveying of dedicated stent systems and could make a contribution to the development of these devices. The proximal diameter and proximal area of the stent system were considerably smaller in human autopsy heart samples than in porcine heart samples and than those given by the manufacturer. Special consideration should be given to the stent deployment procedure (and to the follow-up) of dedicated stent systems, considering final intravascular ultrasound or optical coherence tomography to visualize (and if necessary optimize) stent expansion.     

Effects of dietary fish oil on the depletion of carcinogenic PAH-DNA adduct levels in the liver of B6C3F1 mouse

Many carcinogenic polycyclic aromatic hydrocarbons (PAHs) and their metabolites can bind covalently to DNA. Carcinogen-DNA adducts may lead to mutations in critical genes, eventually leading to cancer. In this study we report that fish oil (FO) blocks the formation of DNA adducts by detoxification of PAHs. B6C3F1 male mice were fed a FO or corn oil (CO) diet for 30 days. The animals were then treated with seven carcinogenic PAHs including benzo(a)pyrene (BaP) with one of two doses via a single intraperitoneal injection. Animals were terminated at 1, 3, or 7 d after treatment. The levels of DNA adducts were analyzed by the ³²P-postlabeling assay. Our results showed that the levels of total hepatic DNA adducts were significantly decreased in FO groups compared to CO groups with an exception of low PAH dose at 3 d (P = 0.067). Total adduct levels in the high dose PAH groups were 41.36±6.48 (Mean±SEM) and 78.72±8.03 in 10⁹ nucleotides (P = 0.011), respectively, for the FO and CO groups at 7 d. Animals treated with the low dose (2.5 fold lower) PAHs displayed similar trends. Total adduct levels were 12.21±2.33 in the FO group and 24.07±1.99 in the CO group, P = 0.008. BPDE-dG adduct values at 7 d after treatment of high dose PAHs were 32.34±1.94 (CO group) and 21.82±3.37 (FO group) in 10⁹ nucleotides with P value being 0.035. Low dose groups showed similar trends for BPDE-dG adduct in the two diet groups. FO significantly enhanced gene expression of Cyp1a1 in both the high and low dose PAH groups. Gstt1 at low dose of PAHs showed high levels in FO compared to CO groups with P values being 0.014. Histological observations indicated that FO played a hepatoprotective role during the early stages. Our results suggest that FO has a potential to be developed as a cancer chemopreventive agent.