Genetic Engineering Approaches to Pig Production*

Modern biotechnology promises a number of new applications in animal breeding and production. Although conventional pig breeding has achieved a high level of efficiency and productivity numerous problems have been encountered with animal health and the loss of meat quality. Selection based on phenotypic performance data of individual animals does not take into account the importance of specific genes and their relevance within a complex regulatory system. In most cases it is therefore difficult to trace back the genetic origins of clinically important disorders. The application of genetic engineering techniques in pig production will facilitate diagnosis, improvement of productivity, and animal health by allowing direct genetic manipulation. Attention must be focussed on the physical and genetic analysis of the procine genome. The isolation and characterisation of genes, DNA-markers, polymorphic DNA-fragments, and their chromosomal assignment will be important prerequisites and tools for the elucidation of genetic disorders. Especially the detection of heterozygous carriers of recessive disorders and their elimination from the breeding stock will increase selection accuracy and decrease the generation intervals. But also the rapid and simple detection of infectious diseases, which is sometimes difficult if not impossible at present, will improve animal health and welfare. Although the production of transgenic animals either by DNA-microinjection into zygotes or the use of embryonal stem cells manipulated in vitro is less straightforward than DNA-based diagnosis it will play an important role in the direct manipulation of the porcine genome and genes. Breeding programmes including the use of transgenic livestock have already been developed. There is no doubt that genetic engineering has reached a degree of practical feasibility, allowing it to play an important role in pig breeding in particular and animal production in general.     

Association of polymorphism of Fc receptor γ chain gene at position-29 in promoter with the susceptibility to systemic lupus erythematosus in southern Chinese

AIM:To detect the association between the polymorphism of Fc receptor γ chain gene at position-29 in promoter and systemic lupus erythematosus(SLE).METHODS:The genotypes at position -29 in promoter of Fc receptor γ chain gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 180 patients with SLE and 140 ethnically matched controls in southern China.RESULTS:The frequencies of TT genotype(33.3%) and T allele (54.4%) at position -29 in patients with SLE were significantly higher than those in controls (17.2% and 42.9%, respectively), whereas, the frequencies of GG genotype (24.4%) and G allele (45.6%) in patients with SLE were remarkably lower than those in controls (31.4% and 57.1%, respectively) (P＜0.05). The TT genotype and T allele at position -29 were not associated with lupus nephritis in SLE patients (P＞0.05).CONCLUSION:Our results indicate that the T allele at position -29 in promoter of Fc receptor gene probably contributes to the susceptibility to SLE, but does not play a role in the occurrence of lupus nephritis.     

徐淮山羊PPAR基因克隆及其表达产物亚细胞定位的研究

研究旨在克隆徐淮山羊过氧化物酶体激活增殖受体(PPAR)基因的cDNA,并通过EGFP融合蛋白对该基因产物亚细胞水平定位。采用RT-PCR方法克隆徐淮山羊脂肪组织PPAR基因cDNA,并构建含有增强型绿色荧光蛋白(EGFP)报告基因的融合表达载体pEGFP-C1-PPAR,聚乙烯亚胺(PEI)介导基因转染NIH-3T3细胞,48h后荧光倒置显微镜下观察基因表达产物的分布,RT-PCR检测mRNA在体外水平的表达。结果表明:首次成功克隆出徐淮山羊PPAR基因的全序列cDNA,大小为1428bp,GenBank登录号为GU082382;构建了融合表达载体pEGFP-C1-PPAR;RT-PCR检测mRNA表达明显;EGFP-PPAR融合蛋白定位在NIT-3T3细胞质中。体外克隆的徐淮山羊PPAR基因cDNA,在单细胞水平上可表达于细胞质中,结果为进一步研究PPAR的生物学功能奠定基础。     

茶色素对胰岛素抵抗大鼠胰岛素受体的影响

探讨茶色素对胰岛素抵抗大鼠红细胞胰岛素受体的影响。将50只清洁级SD大鼠随机分成对照组、高脂组和低中高剂量茶色素干预组。高脂组和茶色素干预组以高脂饲料饲喂8周诱发肥胖致胰岛素抵抗模型,茶色素干预组再给予0.1、0.2g/(kgbw·d)和0.4g/(kgbw·d)剂量茶色素灌胃8周,实验结束后采血测定红细胞胰岛素受体水平,同时测定血糖、血胰岛素、血FFA及血痩素。与对照组相比,高脂组空腹胰岛素、痩素和FFA明显升高(P<0.05),而红细胞高亲和力受体数目明显降低(P<0.05),与高脂组相比,中高剂量茶色素干预组空腹胰岛素、痩素和FFA明显降低(P<0.05),而红细胞高亲和力胰岛素受体数目明显升高(P<0.05)。说明茶色素干预可提高胰岛素抵抗大鼠胰岛素高亲和力受体数目,其机制可能与降低痩素和FFA水平间接提高受体数目或直接作用于受体基因调节元件有关     

温度、丙烯和1-MCP对西洋梨果实乙烯合成和乙烯受体ETR1同源基因表达的影响

研究了贮藏温度、丙烯和 1 MCP (1-甲基环丙烯 )处理对帕斯—卡桑 (Passe -Crassane)西洋梨果实成熟的影响。结果表明 :低温可促进果实的乙烯合成 ,未经低温处理的果实 ,用丙烯处理 ,不能正常合成乙烯。丙烯促进乙烯合成的效应随冷藏时间延长而增强。1 MCP处理可抑制冷藏果实的乙烯合成。定量PCR分析结果表明 ,果实中乙烯受体ETR1同源基因的表达不受低温的调节 ;但冷藏后升温或升温后用丙烯处理时 ,mRNA含量降低     

Protective effect of C1q/tumor necrosis factor-related protein 3 on vascular endothelium in rats with hyperuricemia

AIM To study whether C1q/tumor necrosis factor （TNF）-related protein 3 （CTRP3）protect vascular endothelium in rats with hyperuricemia and its potential mechanisms. METHODS An animal model of hyperuricemia was established by using male SD rats drinking 10% fructose water （n=10）. The rats drinking normal water served as normal controls （n=10）. After 12 weeks, the rats were given a single injection with Ad-CTRP3 or Ad-GFP. The experiment was ended at 14th day after transfection.The serum levels of uric acid and nitric oxide （NO） were evaluated. The serum contents of TNF-α and interleukin-6 （IL-6） were measured by ELISA. HE staining and TUNEL assay were used to assess the morphological changes of intima and apoptosis of endothelial cells in thoracic aorta, respectively. The mRNA levels of endothelial nitric oxide synthase （eNOS）, TNF-α and IL-6 were detected by RT-qPCR. The protein levels of CTRP3 and Toll-like receptor 4 （TLR4） were determined by Western blot. RESULTS Compared with normal control group, the rats with hyperuricemia showed lower CTRP3 and higher TLR4 protein levels in the thoracic aorta （P<0.05）. Hyperuricemic rats had higher serum contents of uric acid, TNF-α and IL-6 （P<0.05）. Also, the intima structure disturbance of thoracic aorta, increased apoptotic rate, higher mRNA levels of TNF-α and IL-6 as well as lower mRNA levels of eNOS were observed （P<0.05）. By contrast, CTRP3 over-expression decreased TLR4 protein levels, reduced inflammatory cytokines, and obviously improved the morphology and function of thoracic aorta in the rats with hyperuricemia. CONCLUSION CTRP3 protect vascular endothelium in rats with hyperuricemia maybe via down-regulation of TLR4- mediated inflammatory signaling pathway.     

环境基准值的科学研究与我国环境标准的修订

对环境基准和环境标准在概念上进行了辨析,论述了环境基准科学研究中需要把握的几个核心问题,其中最为关键的一步是不同类型生态受体的选取,因此环境基准值应该是基于不同保护对象的多目标函数或一个范围值。鉴于我国环境标准迫切需要修订这一实际,呼吁加强环境基准的研究,尤其是要开展一次国家层面上的土壤环境基准的系统研究。     

栀子苷的降糖作用和对PPARγ受体的激活

葡萄糖消耗试验表明,栀子苷能显著促进前脂肪细胞对葡萄糖的吸收。小鼠荷糖试验和四氧嘧啶致糖尿病小鼠血糖试验证实,栀子苷具有剂量依赖性降低小鼠餐后血糖及糖尿病高血糖的作用。通过基于PPARγ受体信号通路的报告基因诱导表达试验,发现栀子苷在体内外的降糖功效,可能与过氧化物酶增殖体激活受体γ的激活有关。     

Effects of mutations in Drosophila nicotinic acetylcholine receptor subunits on sensitivity to insecticides targeting nicotinic acetylcholine receptors

Several strains of Drosophila melanogaster possess mutant alleles in nicotinic acetylcholine receptor (nAChR) subunits, Dα1 and Dβ2 that confer resistance to neonicotinoids such as imidacloprid and nitenpyram, and Dα6, that confers resistance to spinosyns. These mutant strains were bioassayed with a selected set of nAChR active insecticides including neonicotinoids, spinosad, and sulfoxaflor, a new sulfoximine insecticide. All of the neonicotinoids examined, except dinotefuran showed reduced insecticidal efficacy on larvae of the Dα1 mutant, suggesting that this subunit may be important in the action of these insecticides. All of the neonicotinoids, including dinotefuran, showed reduced insecticidal efficacy on larvae possessing the Dβ2 mutation. A similar pattern of broad neonicotinoid resistance to that of Dβ2 alone was also observed for larvae with both the mutations (Dα1 + Dβ2). The Dβ2 mutation exhibited a lower level of cross-resistance to sulfoxaflor (<3-fold) than to any of the neonicotinoids (>13-fold). In contrast, there was no cross-resistance for any of the neonicotinoids or sulfoxaflor in adult flies with the Dα6 mutation, which confers high levels of resistance to spinosad. Thus in the D. melanogaster strains studied, target site resistance observed for the neonicotinoids and the spinosyns does not translate directly to resistance towards sulfoxaflor.     

Expression of natural killer cell stimulatory receptor NKG2D and its ligand MICA in Tibetan patients with gastric cancer at Qinghai plateau

AIM: To investigate the expression of natural killer cell stimulation receptor D(NKG2D) in peripheral blood and the expression of major histocompatibility complex class I chain-related protein A(MICA) on the gastric carcinoma tissue and the neighboring non-cancerous tissue in Tibetan patients with gastric cancer at Qinghai plateau. METHODS: Samples of 33 patients and 20 healthy persons were collected to detect NKG2D in peripheral blood by Flow cytometry analysis. The expressions of MICA in part of the correspondent tissues were examined by RT-PCR and immunohistochemistry.RESULTS: The expression of NKG2D in the plateau of Tibetan patients with gastric cancer group (13.47 ±5.26)% was significantly lower than that in healthy groups (32.62±10.08)%. The mRNA level of MICA in gastric cancer was significantly higher than that in neighboring non-cancerous tissue (P<0.05). The expressions of MICA proteins showed a significant difference between the neighboring non-cancerous tissue (21.21%, 7/33) and the gastric carcinoma tissue (78.79%, 26/33), between the high differentiation (60.00%, 3/5) and the moderate (72.73%, 8/11), low differentiation (88.24%, 15/17). The expressions of MICA were not correlated with tumor size, sex, age, lymph node metastasis of the patients (P>0.05). Spearman rank correlation displayed that the expressions of MICA mRNA were positively correlated with the MICA protein level (r=0.903, P<0.01).CONCLUSION: The immune escape in Tibetan patients with gastric cancer at Qinghai plateau probably relates to the down-regulation of NKG2D and the up-regulation of its ligand MICA. The activity of NK cells and the anti-cancer cellular immunity level are descending in the plateau of Tibetan patients with gastric cancer. The decrease in the receptor NKG2D is a reason for the activity of NK cell descending.