鸡软骨细胞体外分离培养鉴定以及过表达miR-15a对软骨细胞的影响

为分析miR-15a在肉鸡不同组织中的表达情况,并探究过表达miR-15a对体外培养鸡软骨细胞的影响。本研究首先通过倒置显微镜观察、PCR、凝胶电泳和甲苯胺蓝染色对体外分离培养的软骨细胞进行鉴定。通过实时荧光定量PCR检测miR-15a在肢体内外翻畸形(valgus-varus deformity, VVD)组和健康组肉鸡中(各3只)各组织的表达量。之后通过CCK8和EDU方法分析软骨细胞过表达miR-15a后细胞增殖情况。软骨细胞转染miR-15a mimics后,通过qPCR检测软骨细胞的标志基因Collagen-2、Aggrecan、Collagen-10,成熟分化基因Runx2、Sox9、VEGF、MMP9,炎性因子IL-1β、IL-6、IL-8、IL-10、TNF-α、TGF-β3以及凋亡基因Fas、FasL、Bcl-2的表达量。并构建FKBP5 3'UTR的野生型载体和突变型载体,通过双荧光素酶检测报告检测miR-15a与FKBP5的靶向关系。结果表明,本研究所用的胰蛋白酶、Ⅱ型胶原酶和透明质酸酶联合消化法成功分离得到状态良好的软骨细胞。荧光定量结果显示,miR-15a在各组织中均有表达,与健康组相比,miR-15a在VVD组的肝(P < 0.01)、脾(P < 0.05)、胸腺(P < 0.01)中的表达量显著升高,在心和胸肌组织中的表达量显著降低(P < 0.01)。CCK8与EDU分析结果显示,与NC组相比,过表达miR-15a组软骨细胞增殖速度显著降低(P < 0.01),增殖细胞数量显著减少(P < 0.01)。qPCR结果显示,与mimics NC组相比,miR-15a mimics组的软骨细胞标志基因Aggrecan、成熟分化基因Sox9、Runx2表达量显著降低(P < 0.05),Fas基因表达量极显著上升(P < 0.01),FasL基因和抗凋亡基因Bcl-2极显著下降(P < 0.01)。成功构建了FKBP5 3'UTR野生型和突变型载体,双荧光素酶检测报告结果显示预测的靶基因FKBP5与miR-15a没有靶向关系。本研究成功分离并鉴定了鸡软骨细胞,过表达miR-15a抑制鸡软骨细胞增殖、成熟和分化并促进细胞凋亡。     

GAPB基因原核表达载体的构建

以拟南芥cDNA为模板,用PCR扩增出GAPB的基因全长,然后将GAPB基因片段连接到PET28a（＋）载体上,构建重组质粒并转化大肠杆菌DH5α,经菌落PCR和酶切鉴定筛选出阳性克隆,测序正确后,再将阳性克隆的质粒转化大肠杆菌BL21（DE3）。结果表明：成功构建了原核表达载体PET28a（＋）-GAPB,为后续的G...     

Effect of miR-19a on lipid catabolism in hepatocyte LO2

AIM: To observe the effect of microRNA-19a (miR-19a) on the lipid catabolism of hepatocyte LO2, and to explore the potential mechanism. METHODS: miR-19a was over-expressed or silenced by transfection of miR-19a mimics or miR-19a inhibitor into LO2 cells, then the mRNA level of miR-19a was detected by real-time PCR. The potential target of miR-19a was found by the method of bioinformatics through internet website. The effect of miR-19a on the 3' UTR of peroxisome proliferator-activated receptor α (PPARα) was measured by dual luciferase reporter assay, and the protein level of PPARα and its 2 major downstream rate-limiting enzymes involved in lipid catabolism, acyl-coenzyme a dehydrogenase (ACADM) and carnitine palmitoyltransferase 1A (CPT1A), were detected by Western blotting. Meanwhile, the effect of miR-19a on the generation of ketone body was measured by beta-hydroxybutyric acid (β-OHB) detection assay. RESULTS: The mRNA level of miR-19a was dramatically elevated by the transfection of miR-19a mimics, and sharply decreased by the transfection of miR-19a inhibitor (P<0.05). PPARα was found as a potential target of miR-19a, and dual luciferase reporter assay and Western blotting confirmed the regulatory effect of miR-19a on the expression of PPARα, with the protein level changes of ACADM and CPT1A. miR-19a mimics down-regulated, while miR-19a inhibitor up-regulated the concentration of β-OHB in LO2 cells (P<0.05). CONCLUSION: miR-19a regulates the lipid catabolism of hepatocytes by targeting the PPARα and its 2 downstream rate-limiting enzymes.     

新冠肺炎疫情与猪肉价格波动初探

基于2020年3月-2020年10月猪肉价格及新冠肺炎疫情指数,借助VAR模型,定量研究新冠肺炎疫情对于猪肉价格的影响。研究结果表明:新冠肺炎疫情对猪肉价格存在影响,促使猪肉价格上涨,在此基础上文章提出相应措施。     

Protein and energy concentrations of meat meal and meat and bone meal fed to pigs based on in vitro assays

The objective of this study was to develop equations for estimating ileal digestible crude protein (CP) and metabolizable energy (ME) contents of meat meal (MM) and meat and bone meal (MBM) as feed ingredients for pigs based on in vitro assays. Test ingredients were 4 sources of MM and 3 sources of MBM. Ash and CP contents of the ingredients ranged from 3.8% to 33.1% and 46.8% to 82.9% (as-is basis), respectively. In vitro ileal disappearance (IVID) of CP was determined and ileal digestible CP content was calculated by multiplying CP content by IVID of CP. In vitro total tract disappearance (IVTTD) of dry matter (DM) was determined and ME was calculated using gross energy, CP contents, and IVTTD of DM. The IVID of CP and IVTTD of DM ranged from 77.2% to 88.7% and from 82.7% to 92.4%, respectively. Calculated ileal digestible CP and ME contents ranged from 37.8% to 73.5% DM and 2,405 to 3,905 kcal/kg DM, respectively. Ash contents were negatively correlated (P < 0.001) with CP (r = −0.99), in vitro ileal digestible CP (r = −0.97), gross energy (r = −1.00), in vitro digestible energy (r = −0.97), and adjusted ME (r = −0.97). The most fitting equations for ileal digestible CP and adjusted ME were: ileal digestible CP (% DM) = 11.91 − 0.90 × Ash (% DM) + 0.74 × IVID of CP (%) (R² = 0.99) and adjusted ME (kcal/kg DM) = 130.85 − 50.90 × ash (% DM) + 47.06 × IVTTD of DM (%) (R² = 0.99). To validate the accuracy of the prediction equations for ME, mean bias and linear bias were determined using a regression analysis. Calculated ME values of MM and MBM were in a good agreement with data obtained from animal experiments based on a statistically insignificant bias in the models. In conclusion, ME concentrations of MM and MBM as swine feed ingredients can be calculated using ash concentration and in vitro disappearance of dry matter.     

Experiment for Condensing Heat Transfer Performance in a New Type of Horizontal Three Dimensional Inner Microfin Tubes

A new type of three dimenstional inner microfin tube is devoloped. Under four different pressures and in the range of condensing mass velocities from 33 to 153 kg/m2 . s,and by the experimental study of the three kinds of tubes with different microfin parameters, the performances of the condensation heat transfer for R11 in the horizontal microfin tubes and obtained. The experiments show that the inner microfin tube is a hopeful and nighty effective condensing heat transfer enhancement tube.     

Site‐specific management is crucial to managing Mikania micrantha

Increasingly, weeds have been taking on global distributions. With the proliferation of invasive weeds has come the challenge of managing these species over broad geographical regions, with diverse habitats and political jurisdictions. Here, we review the management of Mikania micrantha Kunth (Asteraceae; mile‐a‐minute) throughout its invaded range, extending through most of the Pacific islands and southern and south‐east Asia. Context matters when determining the best course of action for managing M. micrantha, as it has invaded a large variety of agricultural and natural systems. In Queensland, Australia and Florida, USA, M. micrantha has been targeted in relatively successful eradication campaigns, highlighting the importance of early detection and rapid response methods, while elsewhere in its invaded range, populations are either still increasing or showing limited signs of decline. An inter‐regional approach to research and management should incorporate successful management strategies employed throughout the invaded range including, but not limited to, chemical and cultural control practices, manual and mechanical control, classical biological control using the rust fungus Puccinia spegazzinii, plant–plant competition and integrated approaches utilising two or more control methods concurrently. Additional knowledge of M. micrantha genetics is required to determine if management approaches could be fine‐tuned for particular populations. Countries bordering the Mekong River formed a network in 2011 to co‐ordinate the management of invasive species such as M. micrantha. Expanding such a collaborative approach to other regions could further reduce populations of M. micrantha and limit its spread.     

蔬菜一年三熟高效种植模式分析

绿色有机旱作农业是山西农业的传统特色。长治市属于典型的雨养农业、旱作农业区,长治市潞州区抓住绿色有机旱作农业的发展机遇,积极探索适宜该地区、具有区域特色的绿色有机旱作农业技术模式。该技术模式是通过合理的茬口安排,实行一年多茬栽培,“春白菜-甘蓝-秋白菜”是典型的一年三熟种植模式,通过对这种三收高效种植模式的种植方式、田间管理、病虫害防治等方面的分析,提出了实现高效种植的基本措施;按照绿色有机旱作标准制定种植方案,实现提质增效,通过提高复种指数,增加经济效益,为干旱地区绿色有机旱作农业发展提供技术参考。