Expression of IL-6 mRNA in endometrium with endometriosis

AIM: The purpose of the present study was to explore the relationship between interleukin-6 mRNA expression and endometriosis. METHODS: Using the rat model, IL-6 mRNA expression in the endometrium was examined by RT-PCR. RESULTS: The expression of IL-6 mRNA in control rats did not change at 2, 4, 6 and 8 weeks after sham operation (P>0.05), but in model rats it gradually increased at 2, 4, 6 and 8 weeks after endometriosis (P<0.01). The expression of IL-6 mRNA in uterine endometrium with endometriosis was lower than in endometriotic tissue, but higher than in endometrium from healthy controls. CONCLUSION: The IL-6 mRNA expression may contribute to the development of endometriosis . The increase in IL-6 mRNA expression may promote the implantation and growth of endometriotic tissue.     

两种一步法RNA提取试剂的比较

RNA的提取在生物医学研究和临床检测上应用广泛。本文分别用进口Invitrogen公司生产的以及我们自己研制的一步法RNA提取试剂，提取大肠杆菌RNA、鸡肌肉组织RNA和尿囊液RNA，通过核酸电泳和荧光定量RT-PCR进行比较。结果表明用我们自己研制的和进口的试剂提取的RNA都不含有DNA；这些RNA都可以作为模板进行RT-PCR检测，并且在数量上相互之间没有明显差异。说明我们自己研制的一步法RNA提取试剂可以取代进口试剂。此外，本文还讨论了此类试剂一些评定方法。     

适用于转录组测序的大白菜小孢子总RNA提取方法筛选

为了找到适用于转录组测序要求的大白菜小孢子总RNA 提取方法，以出胚率较高的吉红82 为试材，以热激诱导处理后的小孢子为研究对象，比较分析了TRIzol 法、改良CTAB 法及超纯试剂盒法3 种方法的RNA 提取效果。结果表明：TRIzol法（磨样）和改良CTAB 法提取的RNA 浓度较高，但完整性较差；超纯RNA 提取试剂盒法提取的RNA 质量最高，且完整性好，能够满足转录组测序的要求。     

基于紫花苜蓿混播草地的全草型肉羊放牧育肥模式案例研究

本研究旨在探讨紫花苜蓿（Medicago sativa L.）混播草地放牧利用的生产潜力和集成全草型集约化肉羊放牧育肥技术模式。2017-2018年，在位于河北省廊坊市的中国农业科学院国际农业高新技术产业园内开展了2年羔羊育肥的划区轮牧试验，研究了紫花苜蓿混播草地的地上生物量、育肥羊增重以及羊肉品质的变化规律。结果表明：混播草地地上生物量超过11.2 tDM·hm^-2，5月份紫花苜蓿混播草地的粗蛋白为20.92%，每公顷紫花苜蓿混播草地可满足51只育肥羊、放牧育肥150天，每只增重30 kg所需的干物质、代谢能和粗蛋白三个方面的全部需求。与此同时，紫花苜蓿混播草地还可以显著增加羊肉中欧米伽3型多不饱和脂肪酸的含量，n-6/n-3比例为2.72，羊肉健康品质显著提高。以紫花苜蓿混播草地为基础的全草型肉羊放牧育肥技术具有广阔的应用前景，对我国苜蓿产业以及草地畜牧业发展具有重要意义。     

Effect of the particle size on the ammonia removal rate and the bacterial community composition of bioflocs

The total ammonia nitrogen (TAN) removal efficiency and bacterial community composition of bioflocs with <50-μm particle size, > 50-μm particle size and un-sieved bioflocs were investigated in the current study. The initial ratio of dissolved organic carbon to TAN (DOC/TAN) in the three groups were about 14:1. No significant difference was found in the removal rate of TAN, average concentrations of TAN and nitrite nitrogen among the three groups (P > 0.05). The C/N (w/w) ratio of the > 50-μm bioflocs was significantly higher than those of the other groups. No significant differences were found in the crude protein content in the bioflocs among the three groups. The development of the bacterial community compositions of the bioflocs was analyzed by Illumina MiSeq sequencing analyses. Most OTUs were shared among the three groups at all the sampled time points. With the increase in the relative abundance of phylum Firmicutes, that of phylum Proteobacteria, Chorolexi, and Bacteroidetes decreased in all the three groups. The phylum Firmicutes and genus Bacillus were predominant in all the sampled time points. At the end of the experiment, genus Bacillus accounted for 81% in the < 50-μm group, 82% in the > 50-μm group, and 75% in the un-sieved group.     

油菜种子低亚油酸和亚麻酸含量异交稳定技术的建立

为降低油菜种子中的亚油酸和亚麻酸含量并使之在异交条件下保持基本稳定,以高油品种CY2作为受体亲本,采用RNA干涉技术沉默油菜内源Bn FAD2基因的表达,抑制油酸脱饱和反应。育成的3个独立转基因株系的亚油酸和亚麻酸含量下降到6%以下,其中亚油酸含量较受体亲本降低78.2%~86.5%,亚麻酸含量降低53.4%~65.8%。将3个转基因株系与2个亚油酸和亚麻酸含量较高的常规品种正反交,F1种子中这2种脂肪酸的含量依然保持在与转基因亲本相仿的低水平。由此可知,通过RNA干涉技术沉默Bn FAD2基因的表达可赋予油菜低亚油酸和亚麻酸含量的异交稳定特性。这一特性的获得对于今后培育异交稳定的高油酸油菜新品种具有重要意义。     

Effects of Smo gene silencing on cell activity and apoptosis of human cervical carcinoma HeLa cells

AIM: To investigate the effect of Hedgehog (Hh) signaling pathway on the viability and apoptosis of cervical carcinoma cells by shRNA technique to knock down Smoothened (Smo) gene. METHODS: Smo shRNA was used to transfect the cervical carcinoma HeLa cells. The expression of Smo and Gli1 at mRNA and protein levels in the HeLa cells was determined by RT-PCR and Western blot, respectively. The effect of Smo gene silencing on the growth of the cells was measured by MTT assay. The apoptosis and cell cycle were determined by flow cytometry. RESULTS: Compared with control group, the mRNA and protein expression of Smo and Gli1 were evenly reduced obviously after transfected with Smo shRNA for 72 h (P<0.05). The viability of HeLa cells transfected with Smo shRNA was significantly inhibited. The percentages of the cells in G₀/G₁ phase and early apoptosis rate were obviously higher in Smo shRNA transfection group than those in control group. CONCLUSION: Smo gene silencing effectively inhibits the cell growth and induces the apoptosis of human cervical carcinoma cells.     

Sugar beet rotation effects on soil organic matter and calculated humus balance in Central Germany

In order to quantify the influence of land use systems on the level of soil organic matter (SOM) to develop recommendations, long-term field studies are essential. Based on a crop rotation experiment which commenced in 1970, this paper investigated the impact of crop rotations involving increased proportions of sugar beet on SOM content. To this end, soil samples were taken in 2010 and 2012 from the following crop rotation sequences: sugar beet–sugar beet–winter wheat–winter wheat (SB–SB–WW–WW = 50%), sugar beet–sugar beet–sugar beet–winter wheat (SB–SB–SB–WW = 75%), sugar beet–grain maize (SB–GM = 50%) and sugar beet-monoculture (SB = 100%); these were analysed in terms of total organic carbon (TOC) and microbial biomass carbon (MBC) content, MBC/TOC ratio and the TOC stocks per hectare. In addition, humus balances were created (using the software REPRO, reference period 12 years) in order to calculate how well the soil was supplied with organic matter. In the field experiment, harvest by-products (WW and GM straw as well as SB leaves) were removed. After 41 years, no statistically significant differences were measured between the crop rotations for the parameters TOC, MBC, MBC/TOC ratio and the TOC stock per hectare. However, the calculated humus balance was significantly affected by the crop rotation. The calculated humus balance became increasingly negative in the order SB–SB–WW–WW, SB–SB–SB–WW, SB monoculture and SB–GM, and correlated with the soil parameters. The calculated humus balances for the reference period did not reflect the actual demand for organic matter by the crop rotations, but instead overestimated it.     

Bacterial community analysis of purulent material from liver abscesses of crossbred cattle and Holstein steers fed finishing diets with or without tylosin

Liver abscesses in feedlot cattle are polymicrobial infections. Culture-based studies have identified Fusobacterium necrophorum as the primary causative agent, but a number of other bacterial species are frequently isolated. The incidence of liver abscesses is highly variable and is affected by a number of factors, including cattle type. Holstein steers raised for beef production have a higher incidence than crossbred feedlot cattle. Tylosin is the commonly used antimicrobial feed additive to reduce the incidence of liver abscesses. The objective of this study was to utilize 16S ribosomal RNA amplicon sequence analyses to analyze the bacterial community composition of purulent material of liver abscesses of crossbred cattle (n = 24) and Holstein steers (n = 24), each fed finishing diet with or without tylosin. DNA was extracted and the V3 and V4 regions of the 16S rRNA gene were amplified, sequenced, and analyzed. The minimum, mean, and maximum sequence reads per sample were 996, 177,070, and 877,770, respectively, across all the liver abscess samples. Sequence analyses identified 5 phyla, 14 families, 98 genera, and 102 amplicon sequence variants (ASV) in the 4 treatment groups. The dominant phyla identified were Fusobacteria (52% of total reads) and Proteobacteria (33%). Of the top 25 genera identified, 17 genera were Gram negative and 8 were Gram positive. The top 3 genera, which accounted for 75% of the total reads, in the order of abundance, were Fusobacterium, Pseudomonas, and Bacteroides. The relative abundance, expressed as percent of total reads, of phyla, family, and genera did not differ (P > 0.05) between the 4 treatment groups. Generic richness and evenness, determined by Shannon–Weiner and Simpson’s diversity indices, respectively, did not differ between the groups. The UniFrac distance matrices data revealed no clustering of the ASV indicating variance between the samples within each treatment group. Co-occurrence network analysis at the genus level indicated a strong association of Fusobacterium with 15 other genera, and not all of them have been previously isolated from liver abscesses. In conclusion, the culture-independent method identified the bacterial composition of liver abscesses as predominantly Gram negative and Fusobacterium as the dominant genus, followed by Pseudomonas. The bacterial community composition did not differ between crossbred and Holstein steers fed finishing diets with or without tylosin.     

Differentially expressed genes identified through RNA‐seq with extreme values of principal components for beef fatty acid in Nelore cattle

The aim of this study was to identify differentially expressed genes (DEG) in the Longissimus thoracis muscle of Nelore cattle related to fatty acid (FA) profile through RNA sequencing and principal component analysis (PCA). Two groups of 10 animals each were selected containing PC1 and PC2 extreme DEG values (HIGH × LOW) for each FA group. The intramuscular fat (IMF) was compared between cluster groups by ANOVA, and only the sum of monounsaturated FA (MUFA) and ω3 showed significant differences (p < .05). Interestingly, the highest percentage (95%) of phenotypic variation explained by the sum of the first two PC was observed for ω3, which also displayed the lowest number of DEG (n = 1). The lowest percentage (59%) was observed for MUFA, which also revealed the largest number of DEG (n = 66). Since only MUFA and ω3 exhibited significant differences between cluster groups, we can conclude that the differences observed for the remaining groups are not due to the percentage of IMF. Several genes that have been previously associated with meat quality and FA traits were identified as DEG in this study. The functional analysis revealed one KEGG pathway and eight GO terms as significant (p < .05), in which we highlighted the purine metabolism, glycolytic process, adenosine triphosphate binding and bone development. These results strongly contribute to the knowledge of the biological mechanisms involved in meat FA profile of Nelore cattle.