鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

Utilizing a DUALEM‐421 and inversion modelling to map baseline soil salinity along toposequences in the Hunter Valley Wine district

In the oldest commercial wine district of Australia, the Hunter Valley, there is the threat of soil salinization because marine sediments underlie the area. To understand the risk requires information about the spatial distribution of soil properties. Electromagnetic (EM) induction instruments have been used to identify and map the spatial variation of average soil salinity to a certain depth. However, soils vary with depth dependent on soil forming factors. We collected data from a single‐frequency and multiple‐coil DUALEM‐421 along a toposequence. We inverted this data using EM4Soil software and evaluated the resultant 2‐dimensional model of true electrical conductivity (σ – mS/m) with depth against electrical conductivity of saturated soil pastes (EC_p – dS/m). Using a fitted linear regression (LR) model calibration approach and by varying the forward model (cumulative function‐CF and full solution‐FS), inversion algorithm (S1 and S2), damping factor (λ) and number of arrays, we determined a suitable electromagnetic conductivity image (EMCI), which was optimal (R² = 0.82) when using the full solution, S2, λ = 3.6 and all six coil arrays. We conducted an uncertainty analysis of the LR model used to estimate the electrical conductivity of the saturated soil‐paste extract (EC_e – dS/m). Our interpretation based on estimates of EC_e suggests the approach can identify differences in salinity, how these vary with parent material and how topography influences salt distribution. The results provide information leading to insights into how soil forming factors and agricultural practices influence salinity down a toposequence and how this can guide soil management practices.     

Comparative effect of advanced soy products or corn protein concentrate with porcine meal on growth,body composition,and distal intestine histology of Florida pompano,Trachinotus carolinus

The present study was designed to investigate the effects of diets containing advanced soy products (enzyme‐treated soy and fermented soy) or corn protein concentrate (CPC) in combination with porcine meal (PM) to completely replace poultry byproduct meal (PBM) on growth performance, body composition, and distal intestine histology of Florida pompano, Trachinotus carolinus. Four experimental diets were formulated to be isonitrogenous and isolipidic, to contain 400 g/kg crude protein and 80 g/kg lipid. A reference diet (PBM diet [PBMD]) contained 150 g/kg PBM and 495 g/kg soybean meal (SBM), and three test diets were formulated replacing PBM with 15 g/kg of CPC (CPC diet [CPCD]) or replacing all SBM and PBM with 535 g/kg fermented soy (fermented soybean meal diet [FSBMD]) or 451.3 g/kg enzyme‐treated soy (enzyme‐treated soybean meal diet [ESBMD]). All three test diets were supplemented with 38 g/kg of PM. Diets were fed based on a percentage of bodyweight adjusted after sampling the fish every 2 weeks to triplicate groups of Florida pompano juveniles (mean weight 8.06 ± 0.22 g). After 8 weeks of feeding, fish fed CPCD and ESBMD performed equally well in terms of final body weight, thermal growth coefficient, and percentage weight gain in comparison to fish fed PBMD. In all cases, feeding FSBMD resulted in poor feed conversion and lower feed intake compared to other treatments. Protein retention efficiency, whole‐body proximate composition, phosphorus, sulfur, potassium, magnesium, calcium, sodium, and zinc contents were not significantly influenced by the dietary treatments. The results obtained in the present histological study showed no significant differences in the thickness of serous layer, muscular layer, and submucosal layer of the intestine among treatments. Fish fed CPCD showed a significant widening of the lamina propria with an increase of cellular infiltration and higher presence of goblet cells compared to other dietary treatment. Based on these results, 451 g/kg ESBM or combination of 150 g/kg of CPC and 495 g/kg SBM supplemented with 38 g/kg PM can be utilized to develop a practical diet for juvenile Florida pompano without impacting growth, nutritive parameters, and several distal intestine health parameters.     

Replacement of fish oil in plant‐based diets for Pacific white shrimp,Litopenaeus vannamei,by stearine fish oil and palm oil

Stearine fish oil (SFO) and palm oil (PO) have emerged as promising alternatives for the replacement of fish oil (FO) in aquafeeds. This study evaluated the replacement of FO with alternative oils in practical diets for Litopenaeus vannamei. In a clear brackish water study (14.1 g/L) utilizing shrimp (0.29 ± 0.02 g, initial weight), FO was replaced by SFO at inclusion ratios of 100:0, 75:25, 50:50, 25:75, and 0:100 (FO:SFO) and PO as 90% of FO. After 55 days, no significant differences (p < 0.05) in final weight, growth, or survival of shrimp were observed. A second trial (8 weeks) in low‐salinity water (2.1 g/L) with shrimp (0.92 ± 0.02 g, initial weight) evaluated diets with 100% FO, 100% SFO, 90% PO, 90% soybean oil (SO), or 90% flaxseed oil (FXO) as a replacement for FO and four commercially produced diets with 2% of FO, SO, PO, or FXO. One treatment received half rations of the commercial FO diet, and one treatment was based entirely on natural productivity. Results show that the fatty acid profiles of the tail muscle conformed to the lipids of the feed, and highly unsaturated fatty acids (HUFAs) were preserved. Following 8 weeks of culture, there were no significant differences in production performance.     

Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens

We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.     

Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.     

Influence of soil phosphorus status,texture, pH and metal content on the efficacy of amendments to pig slurry in reducing phosphorus losses

Cost‐effective strategies for using chemically amended organic fertilizers need to be developed to minimize nutrient losses in surface and groundwater. Coupling specific soil physical and chemical characteristics with amendment type could increase their effectiveness. This study investigated how water‐extractable phosphorus (P) was affected by chemical amendments added to pig slurry and how this effect varied with soil properties. A 3‐month incubation study was conducted on 18 different mineral soils, stored at 10 °C and 75% humidity and treated with unamended and amended slurry which was incorporated at a rate equivalent to 19 kg total P (TP )/ha. The amendments examined were commercial‐grade liquid alum, applied at a rate of 0.88:1 [Al:TP ], and commercial‐grade liquid poly‐aluminium chloride (PAC ), applied at a rate of 0.72:1 [Al:TP ]. These amendments were previously identified by the authors as being effective in reducing incidental losses of P. The efficacy of the amendments varied with the soil test P, the degree of P saturation (DPS ) and the Mehlich aluminium, iron and calcium, but not soil texture. Chemical amendments were most effective in soils with DPS over approximately 20%. Due to their high cost, the incorporation of amendments into existing management practices can only be justified as part of a holistic management plan where soils have high DPS .     

T7 RNA多聚酶基因的克隆、序列测定及其在E.coli中的表达

从BL21(DE3)E．coli菌株中以PCR的方法扩增得到了与T7RNA多聚酶(T7RNApolymerase，T7pol)基因大小一致的DNA片断。将PCR产物纯化后直接克隆到pGEM—T载体中，经酶切鉴定和DNA序列分析表明克隆得到了正确的T7pol基因。将T7pol基因亚克隆入pET-28b( )中，构建得到原核表达质粒pET28T7。该质粒的BL21(DE3)pLysS转化菌在IPTG的诱导下可表达约98800的蛋白，这与T7pol的相对分子质量一致。将该质粒转化DH5α、JMl09、HBl01、BL21(DE3)和BL21(DE3)pLysS等5种不同的宿主菌，仅有转化T7pol酶活性受到抑制的宿主菌BL21(DE3)pLysS才能得到转化子，而其余4种T7T7pol酶活性不受抑制的E．coli宿主菌不能得到转化子。pET28T7原核表达质粒这种仅能在T7pol酶活性受到抑制的宿主菌中才能存活的现象说明本试验所克隆的T7pol基因能正确表达出具有RNA转录酶活性的蛋白。     

硼浓度对毛木耳菌丝生长的影响试验

试验通过不同浓度的Na2B4O7在改变PDA培养基及棉子壳一木屑培养基的试验,测定了其对毛木耳菌丝生长的影响。结果表明:在200mg／kg浓度下促进作用最为显著,而其浓度超过400 mg／kg时对菌丝有强烈的抑制作用。