基于转录组学的白扁豆总皂苷抑制前列腺癌细胞系PC-3细胞生长的机制研究

目的 白扁豆总皂苷是中药白扁豆经过提取分离纯化步骤制备得到,关于白扁豆的总皂苷成分如何影响前列腺癌细胞系PC-3细胞的生长情况缺少研究。因此,有必要探讨白扁豆总皂苷对前列腺癌细胞系PC-3细胞生长的机制研究。方法 本研究采用CCK8方法检测不同浓度的白扁豆总皂苷对前列腺癌细胞系PC-3细胞生长的影响。利用转录组学分析白扁豆总皂苷抑制前列腺癌细胞系PC-3细胞生长的分子机制,并且进一步通过实时定量PCR（qRT-PCR）和蛋白质免疫印迹（Western blot）实验对相关差异基因的表达进行验证。利用Western blot和CCK8检测白扁豆总皂苷处理过表达醛脱氢酶7家族成员A1（ALDH7A1）的PC-3细胞存活率。结果 随着白扁豆总皂苷浓度升高,前列腺癌细胞PC-3的存活率显著下降,白扁豆总皂苷的IC₅₀值为1 086 mg/L。转录组学测序结果显示,与对照组相比,白扁豆总皂苷处理的细胞中有2 360个差异表达基因,其中1 982个基因上调,378个基因下调。基因功能注释（GO）结果显示,差异表达基因显著富集到与有丝分裂纺锤体检查点（mitotic spindle checkpoint）、纺锤体组装检查点（spindle assembly checkpoint）等一系列跟癌症的发生发展密切相关的生物学过程。此外,基因组京都百科全书（KEGG）分析结果也显示,差异表达基因富集在肿瘤代谢等信号通路。进一步对其中的差异基因进行验证,结果显示,与对照组相比,白扁豆总皂苷处理的前列腺癌细胞中ALDH7A1、甘氨酸C-乙酰转移酶（GCAT）和磷酸甘油酸变位酶家族成员4（PGAM4）的蛋白质表达水平明显降低（P<0.05）,而二甲基甘氨酸脱氢酶（DMGDH）和胱硫醚β合成酶样（CBSL）的蛋白质表达水平显著升高（P<0.001）。体外细胞实验结果表明,白扁豆总皂苷通过下调前列腺癌细胞中ALDH7A1的表达抑制PC-3细胞生长。结论 白扁豆总皂苷可能通过下调ALDH7A1表达从而在体外抑制前列腺癌细胞的生长。     

Effect of antioxidants resveratrol and quercetin on in vitro evaluation of frozen ram sperm

The objective was to assess the effects of the antioxidants resveratrol and quercetin on frozen-thawed ram sperm. Semen samples (which exceeded minimum standards) from four mature crossbreed Santa Inês rams were pooled and aliquots of each pool were diluted in Tris-egg yolk-glycerol, with the addition of 0, 5, 10, 15, and 20 μg/mL of resveratrol and quercetin in Experiment 1 and Experiment 2, respectively. In Experiment 1, the proportion of sperm with a high mitochondrial membrane potential was greater (P < 0.02) in the control group than in resveratrol 20 μg/mL group. In Experiment 2, the proportion of sperm with high mitochondrial membrane potential was greater in the control group (P < 0.0001) than in the other experimental groups, and greater in the quercetin 5 μg/mL group (P < 0.05) than in the other quercetin-treated groups. Thus, addition of 5 to 20 μg/mL of either resveratrol or quercetin to the Tris-egg yolk-glycerol extender reduced sperm mitochondrial membrane potential.     

秸秆和地膜覆盖对渭北旱作玉米农田土壤氮组分与产量的影响

基于田间定位试验,研究了秸秆和地膜覆盖措施对旱作春玉米田土壤氮组分和作物产量的影响。试验包括无覆盖对照,秸秆覆盖和地膜覆盖3个处理,观测指标包括土壤全氮(STN)、颗粒有机氮(PON)、潜在可矿化氮(PNM)、微生物量氮(MBN)、硝态氮(NO_3~--N)、铵态氮(NH_4~+-N)含量及作物产量。结果表明:试验进行5到7年后,与对照相比,秸秆覆盖处理0—10 cm土层STN、PON、PNM、MBN和NO_3~--N含量3年平均分别提高了13.11%、64.29%、17.51%、16.94%和55.37%,10—20 cm土层STN、PON、MBN和NO_3~--N含量3年平均分别提高了5.93%、33.33%、15.78%和27.57%(P0.05)。而地膜覆盖处理0—10 cm和10—20 cm土层NO_3~--N的含量较对照分别提高了189.14%和135.75%(P0.05),其他氮组分与对照处理差异不显著。秸秆和地膜覆盖处理玉米产量较对照处理3年平均分别提高了6.90%和36.74%(P0.05)。玉米产量与0—20 cm土层NO_3~--N含量和NO_3~--N/STN值呈显著正相关关系。总的来看,秸秆覆盖能显著增加旱地土壤全氮和活性有机氮含量,促进氮素固定,但需注意作物生长后期补充氮肥以满足作物生长需要。而地膜覆盖能显著提高土壤氮素有效性和作物产量,但不利于土壤有机氮的固定,且表层土壤存在潜在氮淋失风险。     

A STUDY OF THE EFFECTS OF STATIC AND EXTREMELY LOW FREQUENCY MAGNETIC FIELDS ON LIPID PEROXIDATION PRODUCTS IN SUBCELLULAR FIBROBLAST FRACTIONS

Cultured fibroblasts isolated from murine livers by tissue trypsinization were exposed to a static magnetic field (0.490 T) and to extremely low frequency (ELF) magnetic field (50 Hz, 0.020 T). The cultures were exposed to magnetic fields on four consecutive days for exposure times of 2, 4, 8, 16, 32, and 64 min. After such exposures and obtaining of fibroblast subcellular fractions, lipid peroxidation product—malondialdehyde (MDA) was measured. Increased peroxidation of fibroblasts' membrane structures exposed to an ELF magnetic field was observed in subcellular fractions—microsomal, mitochondrial, and nuclear. No changes in peroxidation of membrane structures were found in fibroblasts exposed to a static magnetic field.     

Chemical partitioning of iron,cadmium, nickel and chromium in contaminated soils of south-eastern Nigeria

Abstract

Selected heavy metals Fe, Cd, Ni and Cr were studied in contaminated soil samples collected from south-eastern Nigeria. Geochemical differentiation into different chemical fractions, using Ma and Rao six-step sequential chemical extraction procedure, was carried out to assess the potential mobility and bioavailability of the heavy metals in the soil profiles. The residual fraction was the most important phase for the four heavy metals with the following average percentage values 74.43 for Fe, 37.69 for Cd, 70.11 for Ni and 62.47 for Cr. The carbonate fraction contained an appreciable proportion of Fe, Cd and Ni with the average percentage values of 16.29, 14.86 and 10.47 respectively, while organic fraction was of next importance for Cr with an average percentage value of 27.14. The Fe-Mn oxide fraction also contained 15.86% of Cd. Relatively low amounts of the metals were associated with water soluble and exchangeable fractions. The mobility factors for the metals in all the sites ranged from 8.55 to 40.04 for Fe, 8.66 to 56.58 for Cd, 12.74 to 30.19 for Ni and 0.82 to 7.22 for Cr. The generally low values of mobility factors coupled with significantly high levels of association of the metals with the residual fraction, indicate that the metals do not pose any environmental risk nor hazard.     

黑河下游湿地土壤有机氮组分剖面的分布特征

结合野外调查,用Bremner法研究了黑河下游湿地不同土壤类型的有机氮组分,结果表明:在0—50 cm土层,5种土壤有机氮均以酸解性氮为主,占全氮的71.04%—81.79%。泥炭土、沼泽土、草甸土、亚高山草甸土所含的酸解氮、非酸解氮和酸解氮组分氨态氮、氨基酸态氮、氨基糖态氮含量的剖面分布总体上均随土层深度的增加而呈降低趋势,而风沙土却相反,上述有机氮组分呈升高趋势。5种土壤酸解氮及其组分氨态氮、氨基酸态氮、氨基糖态氮占全氮比例的剖面分布总体上均随土层深度的增加而呈降低趋势,而非酸解氮却呈升高趋势。5种土壤酸解未知态氮含量及占全氮比例均在剖面分布上无明显特征。在0—30 cm各相同土层内,5种土壤酸解氮各组分含量及占全氮比例的大小顺序均为氨基酸态氮氨态氮未知态氮氨基糖态氮;而在30—50 cm土层,5种土壤酸解氮各组分含量及占全氮比例的大小顺序均无明显特征。此外,黑河下游湿地土壤干化、沙化过程中,表层0—10 cm土壤有机氮组分含量变化明显,其中土壤氨态氮对生态环境变化最为敏感。     

Identification of specific proteins and glycoproteins associated with membrane fractions isolated from Zea mays L. coleoptiles

The aim of this work was to identify proteins specific for plant cell membranes which could then be used as unique markers. A crude membrane fraction was isolated from corn coleoptiles and separated on non-linear sucrose density gradients. Separation of endoplasmic reticulum (NADH-cytochrome c reductase), mitochondria (cytochrome c oxidase), golgi (inosine diphosphatase), and plasma membranes (N-1-naphthylphthalamic acid-binding) was achieved. The membrane proteins from the gradient fractions were separated using sodium dodecyl sulphate-poly-acrylamide gel electrophoresis and the gels stained with coomassie blue or with concanavalin A/peroxidase to detect glycoproteins. Proteins specific for the various membranes were identified. Five proteins including two glycoproteins were plasma membrane markers. Protoplasts were isolated and iodinated using lactoperoxidase/glucose oxidase covalently attached to beads. Eleven iodinated proteins were found and three of these corresponded to proteins specifically associated with plasma membranes in the density gradients. Two methods for detecting Ca²⁺-binding proteins following sodium dodecylsulphate polyacrylamide gel electrophoresis were employed. The majority of such proteins were found in the endoplasmatic reticulum and one was specific for plasma membranes. In vitro and in vivo phosphorylation of membrane proteins was examined and the majority of proteins phosphorylated were glycoproteins. Two of the phosphorylated proteins (M_r=110,000 and 20,000) were also iodinated on protoplasts and may be part of the plasma membrane ATPases.Abbreviations ER endoplasmic reticulum - IDP inosine diphosphate - NPA N-1-naphthylphthalamic acid     

Controlling contagious agalactia in artificial insemination centers for goats and detection of Mycoplasma mycoides subspecies capri in semen

Many goat artificial insemination (AI) centers in Spain have adopted new measures to control contagious agalactia (CA). To avoid the introduction of male goats carrying mycoplasma organisms subclinically in their external ear canal (auricular carriers) in these centers, two ear swabs and a blood sample are obtained from all candidate animals for polymerase chain reaction (PCR), culture (swabs) and serologic tests to detect the presence of mycoplasmas. In addition, the semen produced at these centers is routinely cultured and PCR tested also to detect the presence of mycoplasmas. One y after the introduction of this program, we tested 48 ear swabs and 24 blood samples from 24 candidates for admission to these AI Centers. Three of these ear swab samples (3/48, 6.25%) scored positive for the presence of mycoplasmas; Mycoplasma agalactiae (Ma) was detected in two samples and Mycoplasma mycoides subsp. capri (Mmc) in one. All animals were serologically negative for Ma. Also, out of 173 semen samples obtained from 137 admitted animals (2 and 3 samples were obtained in 16 and 10 bucks, respectively), one (1/173, 0.56%) was positive for Mmc. Our findings suggest that ear swab and semen samples are useful tools to control CA at AI Centers. The introduction of this program has also resulted in the first detection of Mmc in semen from a naturally infected goat, confirming the ability of this mycoplasma to colonize the reproductive tract of male goats. These results highlight the need to improve control measures in semen producing centers to minimize the risk of CA transmission.     

Genetic diversity of UPASI tea clones (Camellia sinensis (L.) O. Kuntze) on the basis of total catechins and their fractions

Tea leaf catechins and the ratio of dihydroxylated to trihydroxylated catechin fractions were analysed to identify the genetic diversity of 26 UPASI released tea clones. Principal component analysis (PCA) based on regression factor separated tea clones into five groups according to their jats (Jats are region based rays for e.g., Assam, China and Cambod origin) as well as their quality constituents (such as total polyphenols, total catechins, amino acids in the green leaves and liquor characteristics of black tea), particularly the catechins. Group 1 represented medium quality (quality of the final produce) clones, such as UPASI-10, UPASI-12 and UPASI-15 and drought tolerant clones like UPASI-1, UPASI-2, UPASI-9 and UPASI-10. Group 2 contained purely "China" cultivars while group 3 possessed high quality tea cultivars. "Assam" (group 5) teas had the lowest ratio of dihydroxylated to trihydroxylated catechin fractions (1:4) than the "Chinery" (group 2) teas (1:5). This biochemical differentiation indicated that there is a vast genetic diversity in UPASI released tea clones in terms of catechin fractions, even though the majority of them were selected from one tea estate located in the Nilgiris.     

土壤稳定性有机碳组分与优势细菌门类陆向分布及相关性

【目的】为了探究土壤稳定性有机碳组分和优势细菌门类的陆向分布特征及相关性。【方法】本文选择锡林河流域沿着由水及陆的方向依次采集长期性水流、季节性水流、长期性无水流的湿地及旱地土壤,基于国际腐殖物质协会推荐的方法和16SrRNA基因高通量测序技术分别检测土壤稳定性有机碳组分(富里酸、胡敏酸、胡敏素)含量和优势细菌门类的相对丰度,结合Pearson相关性及冗余分析和结构方程模型研究两者的相关性。【结果】三类稳定性有机碳及酸杆菌门、放线菌门、芽单胞菌门呈大致升高的陆向分布趋势,在长期性无水流的旱地土壤中达到峰值;拟杆菌门则呈现降低的陆向分布趋势。结果显示,芽单胞菌门、酸杆菌门和放线菌门作为长期性无水流旱地土壤的优势细菌门类与胡敏酸、胡敏素含量存在显著(P<0.05)或极显著(P<0.01)正相关关系;拟杆菌门作为长期和季节性水流湿地土壤的优势菌门与富里酸、胡敏酸、胡敏素的含量均呈极显著(P<0.01)负相关关系。结构方程模型结果显示,稳定性有机碳组分与优势细菌门类间存在直接和间接作用。【结论】锡林河流域土壤稳定性有机碳组分及优势细菌门类存在陆向分布特征,稳定性有机碳的升高与...