草鱼3个6-磷酸葡萄糖酶催化亚基的基因表达分析及高糖饲料对其表达的影响

为了探讨6-磷酸葡萄糖酶催化亚基(glucose-6-phosphatase catalytic subunit,G6PC)在草鱼(Ctenopharyngodon idellus)糖代谢中的作用,采用同源序列比对的方式,在草鱼基因组中获取了3个g6pc基因的序列,通过序列比对和进化树分析,将其分别命名为g6pca、g6pcb1和g6pcb2,其编码的氨基酸序列与斑马鱼(Danio rerio)、虹鳟(Oncorhynchus mykiss)和人(Homo sapiens)具有较高的同源性,相似度分别为85%～94%、64%～83%和54%～66%。同线性分析表明g6pc在草鱼染色体上的分布与斑马鱼等高度相似,表明草鱼g6pc基因在进化中具有较高的保守性。利用RT-PCR检测3个基因在鳃、脂肪、脑、心脏、肝、肾、前肠、中肠、后肠和肌肉10个组织中的表达,结果显示, g6pca在脑和肝中表达量较高,脂肪组织次之; g6pcb1在肝中表达量最高,中肠次之; g6pcb2在心脏表达量最高,其次为脂肪组织。同时探讨了高糖饲料对草鱼不同g6pc亚型转录水平的影响,以及不同浓度葡萄糖和胰岛素刺激草鱼肝细胞(L8824)后对g6pca转录水平的影响。结果显示,饲喂高糖饲料(7周)后,与对照组相比,草鱼肝脏g6pca mRNA水平显著升高, g6pcb1和g6pcb2 mRNA水平无显著变化。离体情况,与5 mmol/L葡萄糖组相比,15 mmol/L葡萄糖显著增加了L8824的g6pca mRNA水平,且1 mol/L胰岛素可以抑制这种作用; 30 mmol/L葡萄糖对L8824 g6pca mRNA水平无显著性影响。本研究表明,草鱼g6pc发生加倍后存在功能分化,高糖可以诱导g6pca mRNA的表达,而对g6pcb1和g6pcb2的转录水平无影响,其具体功能还需进一步研究。     

Comparative analysis of genetic effects of wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL

Wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL, carriers of Pm21 and PmV, respectively, confer high resistance to wheat powdery mildew. For better understanding of the difference in genetic effect between them, a RIL population was constructed based on the cross between “Yangmai 18” carrying T6V#2S·6AL and “Yangmai 22” carrying T6V#4S·6DL. Analysis of distribution of the translocations showed that T6V#2S·6AL is much more transmittable than T6V#4S·6DL. By comparing their effects on main agronomic traits, we firstly found that T6V#2S·6AL contributes greatly to top spikelet fecundity, but causes a decrease of 6.7%–10.5% of spike number. No stable effects of T6V#4S·6DL on agronomic traits were found, except for positive effect on plant height. Excitingly, a new recombinant, T6V#4S‐6V#2S·6AL carrying PmV, was screened and proved to have a higher transmission rate than the original translocation T6V#4S·6DL, which will greatly promote the utilization of PmV. The above conclusions of this research will provide important guidance for utilization of Pm21 and PmV more effectively, in wheat powdery mildew resistance breeding.     

Role of TLR4-TRPC6 in LPS-induced NF-κB P65 expression and nuclear translocation in airway epithelial 16HBE cells

AIM: To investigate the role of Toll-like receptor 4 (TLR4) and transient receptor potential channel 6 (TRPC6) signaling pathway in lipopolysaccharide (LPS)-induced nuclear factor-κB (NF-κB) P65 expression and nuclear translocation in airway epithelial cells (16HBE) for supplementing the mechanism for airway inflammation. METHODS: After stimulating the 16HBE cells with LPS at 1 mg/L for 0, 0.5, 2, 6, 12 and 24 h, the expression of NF-κB P65 at mRNA and protein levels in the 16HBE cells were determined by RT-PCR and Western blot respectively, and the nuclear translocation of NF-κB P65 was detected by immunocytochemical staining method. The effects of TLR4 inhibitor CLI-095 at 5 μmol/L and TRPC6 agonist Hyp9 at 10 μmol/L on LPS (1 mg/L)-induced NF-κB P65 expression and nuclear translocation in the 16HBE cells were determined by RT-PCR, Western blot and immunocytochemical staining. RESULTS: LPS increased the mRNA and protein expression of NF-κB P65 and nuclear translocation in the 16HBE cells(P<0.05). TLR4 inhibitor CLI-095 reduced the mRNA and protein expression of NF-κB P65 and nuclear translocation induced by LPS, while Hyp9 enhanced the mRNA and protein expression of NF-κB P65 and nuclear translocation induced by LPS in the 16HBE cells(P<0.05). CONCLUSION: LPS induces the expression and nuclear translocation of NF-κB P65 in the 16HBE cells via TLR4-TRPC6 signaling pathway.     

Molecular cloning and functional characterization of the fatty acid delta 6 desaturase (FAD6) gene in the sea cucumber Apostichopus japonicus

The sea cucumber (Apostichopus japonicus), an important echinodermata, had high value in nutrition and medicine for its rich collagen, sulphated polysaccharide, glycosides and polyunsaturated fatty acids (PUFA). The cDNA of the fatty acid desaturase gene in A. japonicus (AJFAD6) was cloned and was found to encode a desaturase with delta 6 FAD activity. Sequence analysis indicated that AJFAD6 included an open reading frame of 1392 bp, encoding 463 amino acids. AJFAD6 has all the conserved motifs found in other members of the FAD6 family, including an N‐terminal cytochrome b5 domain and three histidine‐rich regions. qRT‐PCR showed that AJFAD6 was expressed in all tissues tested during juvenile development and was mainly expressed in the respiratory tree at 150 days after adherence (150 days) and in the intestine at 100 days. Furthermore, AJFAD6 mRNA was also detected in the analysed adult tissues, with higher expression in the intestine and testis. Functional characterization of AJFAD6 in a recombinant yeast, Pichia pastoris, showed that AJFAD6 could catalyse exogenous linoleic acid (LA) and α‐linolenic acid (ALA) to produce γ‐linoleic acid (GLA) and stearidonic acid (STA), respectively, at conversion rates of 11.1% for LA to GLA and 3.4% for ALA to STA. Our results suggested that the biosynthetic pathway of PUFA existed in the sea cucumber, but endogenous production of eicosapentaenoic acid, arachidonic acid and docosahexaenoic acid from either LA or ALA precursor appeared to be limited.     

Significant improvement of shrimp growth performance by growth hormone‐releasing peptide‐6 immersion treatments

Growth hormone‐releasing peptide‐6 (GHRP‐6) is one of the earliest developed synthetic peptidyl growth hormone secretagogue receptor agonists. These compounds mimic the effect of the endogenous ligand ghrelin. In vertebrates, ghrelin is a potent circulating orexigenic hormone with functional roles in controlling food intake, energy expenditure, adiposity, growth hormone secretion and immunity. Ghrelin has been studied mainly in vertebrates; thus, little is known about its role in invertebrates, including crustaceans. We first evaluated the effect of GHRP‐6 injection over feed intake in shrimp and its effects on shrimp growth when the peptide was administrated by successive immersion baths. GHRP‐6 increased feed intake, body weight and size, the number of rostral spines and gill branches, protein concentration and haemocyte number in treated shrimps. We also evaluated the peptide uptake and clearance in a pharmacokinetics, using [H³]GHRP‐6 administered to postlarvae. Given a limited exposure and efficient clearance of the peptide‐associated radioactivity from larvae, our findings suggested that GHRP‐6‐treated Litopenaeus vannamei can be consumed safely by humans after aquaculture applications. These results propose that GHRP‐6 could be an additional tool to study growth physiology in crustaceans and also a promising candidate for development into a new biotechnology product for improving shrimp growth and quality.     

Association of TNF-α, IL-10 and IL-6 promoter polymorphisms in pulmonary tuberculosis patients and their household contacts of younger age group

Single nucleotide polymorphisms (SNPs) of cytokine genes have been found to be involved in the clinical outcome of Tuberculosis. The present study was aimed to identify the high risk genotypes in Tuberculosis patients and their household contacts. A total of 490 subjects were studied which includes 150 active pulmonary tuberculosis patients (APTB), 190 household contacts (HHC) and 150 healthy controls (HC). The SNPs of TNF-α (-308A/G), IL-10(-1082G/A) and IL-6(-174G/C) were performed by ARMs PCR. The IL-10 GA genotype showed significant association in APTB and HHC and was 2.3 times higher risk in APTB and 3.7 times in HHC compared to HCs. The A allele was found to be significantly associated with the risk of disease. The CC genotype of IL-6 was found to be significantly associated in APTB and an insignificant positive association in HHCs. The multifactor dimensionality reduction (MDR) analysis indicated that the genotypes of IL-6 were showing high risk with GA genotype of IL-10. In conclusion the gene interaction may be useful for identification of genotypes as biomarkers to distinguish high risk individuals.     

不同浓度NAA/6BA配比对不同品种甘薯茎尖培养特性的影响

为明确不同品种甘薯茎尖培养的最佳NAA/6BA配比,设置0.1 mg/L/2.5 mg/L、0.2 mg/L/2.5 mg/L、0.2 mg/L/1.0 mg/L 3组NAA/6BA浓度配比处理,观测不同浓度激素处理对‘北京553’、‘红香蕉’、‘苏薯8号’、‘烟薯25’、‘安吉芋’、‘济徐23’、‘渝紫7号’、‘商薯19’茎尖培养成苗率、愈伤组织直径、不定根数目、叶片数和植株高度的影响,同时调查不同品种试管苗的移栽成活率。结果表明,‘红香蕉’、‘济徐23’、‘渝紫7号’、‘商薯19’在0.1 mg/L/2.5 mg/L的处理下培养效率最高;‘安吉芋’在0.2 mg/L/2.5 mg/L的处理下培养效率最高;‘苏薯8号’、‘北京553’、‘烟薯25’在0.2 mg/L/1.0 mg/L的处理下培养效率最高。因此,在甘薯茎尖培养过程中,不同甘薯品种适宜的NAA/6BA配比存在显著差异,在实际生产中应根据品种特性来进行调整激素的用量和比例。     

保护地黄瓜新品种唐杂6 号的选育

唐杂6 号是以雌性系S16 为母本，以自交系S26 为父本配制的强雌型黄瓜一代杂种。生长势强，商品瓜短棒状， 瓜长12～14 cm，横径4.0～4.3 cm，非特异性环境下雌花率95% 以上，瓜皮嫩绿有光泽，白刺，刺瘤稀小，平均单瓜质量 131.8 g 左右，高抗霜霉病，抗细菌性角斑病，耐白粉病；春保护地栽培平均产量可达8 000 kg·（667 m²） ^-1，秋冬保护地栽 培产量可达6 500 kg·（667 m²） ^-1；适合河北、北京、天津及东北地区春、秋保护地种植。     

Minimally modified low-density lipoprotein up-regulates endothelin receptors in mouse mesenteric artery through p38 MAPK pathway

AIMTo investigate whether minimally modified low-density lipoprotein (mmLDL) affects the quantity and activity of endothelin （ET） type A (ET_A) and type B (ET_B) receptors in mouse mesenteric artery by activating p38 mitogen-activated protein kinase (MAPK) inflammatory pathway. METHODSThe KM mice were divided into normal saline (NS) group (injection of NS via caudal vein), mmLDL group (injection of mmLDL via caudal vein), LDL group (injection of LDL via caudal vein), mmLDL+SB 203580 group (injection of mmLDL via caudal vein and intraperitoneal injection of p38 MAPK pathway specific inhibitor SB 203580) and mmLDL+DMSO group (injection of mmLDL via caudal vein and intraperitoneal injection of DMSO). Mesenteric artery ring segment vasoconstriction dose-response curves affected by sarafotoxin 6c (S6c) and ET-1 were recorded by the myography system. The mRNA levels of ET_B receptor, ET_A receptor and interleukin-6 (IL-6) were detected by RT-qPCR. The protein levels of ET_B receptor, ET_A receptor, IL-6, p38 MAPK, p-p38 MAPK, NF-κB and p-NF-κB were determined by Western blot. The serum concentration of IL-6 was measured by ELISA. RESULTSThe contractile responses of the blood vessel segments to S6c and ET-1 were significantly increased by mmLDL (P<0.01). The mRNA and protein expression levels of ET_A receptor, ET_B receptor, and IL-6 significantly increased (P<0.01). The protein levels of p-p38 MAPK and p-NF-κB were significantly increased (P<0.01). The serum level of IL-6 was significantly increased (P<0.01). These effects of mmLDL were inhibited by p38 MAPK inhibitor SB 203580. CONCLUSION mmLDL increses the serum concentration of IL-6, up-regulates the expression of IL-6, ET_A receptor and ET_B receptor in mouse mesenteric artery, and enhances the vasoconstriction function medi?ated by ET_A and ET_B receptors, which is related to the activation of p38 MAPK inflammatory pathway and downstream NF-κB pathway.     

光响应性阳离子表面活性剂PCDA-C6-NH3+的合成及其对液滴蒸发行为的调控

农药的有效利用与农药液滴在植物叶片上的蒸发过程息息相关。传统表面活性剂虽然可以改变液滴的蒸发模式或速率,但并不能根据外界环境变化对蒸发过程进行主动调节。为实现"自主调控"农药液滴蒸发的目的,本文设计合成了一种含丁二炔官能团的光响应性阳离子表面活性剂(PCDA-C6-NH3+),利用紫外光刺激改变其在液滴中的聚集状态,调控液滴的蒸发过程。该表面活性剂以10,12-二十五碳二炔酸(PCDA)作为骨架,通过酰胺化反应引入末端氨基并质子化后得到。采用紫外-可见光谱研究了PCDA-C6-NH3+的拓扑化学聚合反应,并利用静态表面张力和动态接触角分析了其紫外光照前后的表面活性和蒸发行为。该研究为实现液滴"自主调控"蒸发过程提供了一种新思路,对提高农药利用率具有重要意义。