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Methods seventy-two 15-week-old sd rats were divided into six groups,included control group and five ovariectomized model groups.the model groups were fed with different-dose lycopene[10,15,20 mg/] for 12weeks.then related indexes including length bone mineral density,bone mineral,serum estradiol,sertum alp and uterus index were analyzed.results compared with model group,three different dose groups and nilestriol[1.05mg/] group showed increased bone mineral density,bone mineral,serum estradiol and uterus index,but decreased serum alp (p.05).conclusion as a female sex hormone-like component,lycopene may ameliorate the bone quality and inhibit osteoporosis in ovariectomized rat.

将72只15周龄sd大鼠随机分为6组,空白组做假手术,其余5组做卵巢切除术,术后用药12周后处死,测定右侧股骨骨密度、骨矿含量、血清雌二醇、碱性磷酸酶含量及子宫湿重。结果番茄红素高[20mg/]、中[15mg/]、低剂量组[10mg/]、尼尔雌醇组[1.05mg/kg·bw]能对抗骨密度、骨矿含量、血清雌二醇的下降、血清碱性磷酸酶的升高,与模型组比较,p.05;番茄红素各剂量组、尼尔雌醇组能对抗子宫指数的减轻,与模型组比较,p.05。结论番茄红素可以改善骨质量,抑制大鼠去卵巢骨质疏松的发生。具有类似雌激素的效应。

Result Activity of serum alkalinity phosphatase of cows and sheep was in a normal range. But activity of alkalinity phosphatase of mid pregnancy cows was obviously lower than before pregnancy and not pregnant and later pregnancy (P.05). Alkaline phosphatase retention was lower than 26%. It showed that serum alkalinity phosphatase of cows and sheep came from the skeleton.

结果]所有奶牛和绵羊血清碱性磷酸酶活性均在正常范围内,但奶牛怀孕中期的碱性磷酸酶活性比未怀孕、怀孕前期、怀孕后期明显降低(P.05);碱性磷酸酶保存率均低于26%,表明奶牛和绵羊血清碱性磷酸酶均来自骨骼。

After thecomplete genome extraction of the strain was performed, the genomic DNA was partiallydigested by restriction enzyme Sau3AⅠ, the DNA fragments from 1 to 5Kb was clonedinto prokaryote expression vector pET-28a-c, and transformed host bacteria. The resultsshowed that we succeeded in constructing the gene expression library of haemophilusparasuis serovar 5, which is fundamental for the study of advanced gene screening. Inaddition, primer design was performed based on haemophilus influenzae in this study. In addition, PCR was performed by using genomic DNA of haemophilus parasuisserovar 5 as the template. The results demonstrated that we obtained two neo-gene:23SrRNA gene(conserved gene belonging to the large-subunit of ribosome) and adenylatecyclase gene(encodes adenylate cyclase and participates in converting adenyl nucleosidetriphosphate to cyclic adenosine3",5"-monophosphate). Furthermore, the phylogeneticanalyses between the species was performed, and neighbor-joining tree was constructedbased on comparison of 23S rRNA gene sequences, so it was illuminated betweenHaemophilus parasuis and other species in molecular evolution relationship.

选择我国流行优势菌株副猪嗜血杆菌血清5型地方株为研究对象,提取细菌基因组DNA,用限制性内切酶Sau3AⅠ对基因组DNA进行部分酶切,回收大小为1~5Kb的DNA片段,将其连接入原核表达载体pET-28a-c,最后转化宿主菌,结果成功地构建了基因表达文库,为后续的基因筛选工作奠定基础;另外,本研究选择嗜血杆菌属的流感嗜血杆菌为参考对象进行引物的设计,以副猪嗜血杆菌血清5型地方菌株的基因组DNA为模板,进行PCR扩增反应,结果表明成功地获得两个新基因:23S rRNA基因(存在于核糖体大亚基中的保守性基因)和腺苷酸环化酶基因(负责将腺嘌呤核苷三磷酸转变为环腺苷酸),并进一步做了不同物种之间的分子系统发育分析,构建了基于23S rRNA基因的邻接法系统发育树,阐明了副猪嗜血杆菌与其它菌种的分子进化关系。

Serum alt,ast,and bile acid were observed by enzyme assay and the ratio of ast/alt was worked out. serum bilirubin was measured by peariman and lee method. serum albumin was tested by bromphenol assay and was simultaneously measured to hyaluronic acid, type ⅲ procollagen, laminin and type ⅳ collagen by ria. meanwhile, prothrombin time and platelet count were measured and so on. results: the level of serum aca was raised increasingly in different cirrhotic cases with different classifications of child-pugh, according to severity of the condition.

应用酶联免疫吸附法分别对不同分级的肝硬化患者血清中抗心磷脂抗体的含量进行了测定;用酶法测定了alt、ast与胆汁酸并计算ast/alt值;用peariman和lee改良法测定血清胆红素;用溴甲酚氯比色法测定血清白蛋白;用放射免疫分析法测定透明质酸、ⅲ型前胶原氮端肽、层粘连蛋白与ⅳ型胶原;并测定了凝血酶原时间与血小板计数等相关指标。

The experiment two: enzyme preparation significantly improved average daily gainand feed conversion ratio (P<0.05). Enzyme preparation significantly increased energymetabolizability and digestibility of crude fiber, crude protein and neutral detergent fiber,but had no remarkable effect on digestibility of dry matter, crude fat and acid detergentfiber. Enzyme preparation significantly decreased the relative viscosity of duodenal andjejunal digesta. The pH of intestine had no noticed difference in all groups. Enzymepreparation significantly decreased relative weight of gizzard, proventficulus, duodenum,jejunum and ileum. Enzyme preparation significantly increased villus size of duodenumand jejunum, and villus to crypt ratio of duodenum and ileum significantly increased too.Enzyme preparation considerably decreased ileal crypt height (P<0.05), and didn"t affectthickness of intestinal wall. Supplementing enzyme preparation, the serum glucose, totalprotein and alanine aminotransferase, but enzyme preparation hadn"t noticed influenceupon uric acid, total cholesterol, triglyceride and high-density lipoproteins. Enzymepreparation significantly increased insulin, triiodothyronine and insulin-like growthfactor-Ⅰ. Adding enzyme preparation, the percentage of thyroid stimulating hormone andgrowth hormone in the serum increased 16.44%, 19.18% and 18.84%, 21.74%respectively, and the percentage of glucagon and thyroxine decreased 12.07%, 14.36% and 13.79%, 15.40%, but failed to reach statistical significance (P>0.05). Enzymepreparation significantly increased (P<0.05) the trypsin and amylase activity of duodenaland jejunal digesta, but enzyme preparation didnt affect significantly (P>0.05) theintestinal lipase activity and pancreatic digestive enzyme. Enzyme preparation had nosignificant effect on caecal microbial population.

试验二:酶制剂显著提高平均日增重和饲料转化率(P<0.05);酶制剂显著提高能量代谢率及粗纤维、粗蛋白、中性洗涤纤维消化率(P<0.05),而对干物质、粗脂肪、酸性洗涤纤维消化率影响不显著;酶制剂显著降低十二指肠和空肠食糜相对粘度(P<0.05);添加酶制剂对肠道pH影响不显著;酶制剂显著降低肌胃、腺胃、十二指肠、空肠、回肠相对重(P<0.05),显著提高十二指肠和空肠绒毛高度,显著增加十二指肠和回肠绒毛高度/隐窝深度,降低回肠隐窝深度(P<0.05),对肠壁厚度影响不显著;酶制剂显著提高血清葡萄糖、总蛋白和谷丙转氨酶浓度(P<0.05),对尿酸、总胆固醇、甘油三酯及高密度脂蛋白浓度影响不显著,显著提高胰岛素、T_3、IGF-Ⅰ水平,添加酶制剂后,促甲状腺激素、生长激素分别提高16.44%、19.18%和18.84%、21.74%,胰高血糖素和T_4分别降低12.07%、14.36%和13.79%、15.40%,但差异不显著;酶制剂对胰腺消化酶活性影响不显著,显著增加十二指肠和空肠胰蛋白酶、淀粉酶活性,对小肠脂肪酶活性影响不显著;酶制剂对盲肠微生物菌落数影响不显著。

It showed that the application of 5′-NT in acute hepatitis, live cancer and osteal diseases were significantly higher than that of controls.

用法国Biomerieux公司(5′-NT)诊断试剂盒测定血清5′-NT酶活性,同时测定患者血清谷丙转氨酶和γ-谷氨酰转肽酶,碱性磷酸酶酶活性的变化。

Finally confirmed that,The thighbone and the osteoporosis morbidity is possibly connected protein 6, respectively are: The lactoferrin light chain, membrane association protein A3, the enolase, ATP gather the enzyme, the acetyl coenzyme A reductases, the myo calcium protein; The lumbar vertebra and the osteoporosis morbidity is possibly connected protein 5, respectively are: The actin, the keratin, the enolase, ATP gather the enzyme, the myosin; The thighbone and the strong bone valuable curative effect is possibly connected protein 9, respectively are: The enolase, ATP gather the enzyme, the myo-calcium protein, the creatine activating enzyme isozyme, the phosphoglyceric acid change flavor the enzyme, the myosin, the lactoferrin light chain, the pyruvic acid activating enzyme isozyme, the crown protein; The lumbar vertebra and the strong bone valuable curative effect are possibly connected protein 8, respectively are: The carbonic anhydrase, the actin, the αB-crystal protein, 3-phospho-glycerol aldehyde oxidase, the serum albumin, ATP gather the enzyme, the myosin, the enolase.

最后确认:股骨与骨质疏松发病可能相关的蛋白6个,分别为:乳铁蛋白轻链、膜联蛋白A3、烯醇化酶、ATP合酶、乙酰辅酶A还原酶、肌钙蛋白;腰椎与骨质疏松发病可能相关的蛋白5个,分别为:肌动蛋白、角蛋白、烯醇化酶、ATP合酶、肌球蛋白;股骨与强骨宝疗效可能相关的蛋白9个,分别为:烯醇化酶、ATP合酶、肌钙蛋白、肌酸激酶同工酶、磷酸甘油酸变味酶、肌球蛋白、乳铁蛋白轻链、丙酮酸激酶同工酶、冠蛋白;腰椎与强骨宝疗效可能相关的蛋白8个,分别为:碳酸酐酶、肌动蛋白、αB-晶体蛋白、3-磷酸甘油醛脱氢酶、血清白蛋白、ATP合酶、肌球蛋白、烯醇化酶。

As hollow fiber membranes thickness are different, yet urea clearance rate in simulation solution is relatively high, which can reach more than 64% and little difference among them; the BSA have a relatively high rejection, which can be achieved over 98%. There is a certain difference in the Lysozyme clearance rate, the smaller thickness, the higher the rate of removal, and the removal rate of 10μm thickness of the membrane is 29%. The clearance rates of urea were very high using different fiber diameter dialyzers, at around 80%. Urea clearance ratecan reach to the highest 88.6% using 0.26mm fiber diameter dialyzer. The clearance rates of Lysozyme vary greatly, the smaller the diameter, the higher clearance rate, and the highest removal rate of lysozyme is 62.79% using 0.26mm fiber diameter dialyzer. However, there is little impact on the rejection of BSA, which can reach 98%.

不同壁厚中空纤维膜透析器对模拟液中尿素的清除率均可达到64%以上,且差异不大,对牛血清白蛋白的截留率均可达到98%以上,对溶菌酶的清除率有一定的差异,壁厚越小清除率越高,壁厚为10μm的膜对溶菌酶的清除率为29%;不同纤维内径透析器对尿素的清除率均在80%左右,纤维内径为0.26mm的透析器对尿素的清除率最高,达到88.6%,对溶菌酶的清除率差别很大,内径越小,清除率越高,内径为0.26mm的透析器对溶菌酶的清除率达到62.8%,对牛血清白蛋白的截留影响很小,截留率都能达到98%以上。

Using centrifugal casting method, hollow fiber membrane is encapsulated into dialyzers, and prepared by the water, urea, Lysozyme and BSA simulated liquid form, which uses Lysozyme instead of β_2-MG, BSA instead of human serum albumin. In this paper, the simulation solution is used to evaluate dialysis performance of PES hollow fiber membrane dialyzers.

采用离心浇铸法将中空纤维膜封装成透析器,用溶菌酶代替β_2~-微球蛋白,用牛血清白蛋白代替人血清白蛋白,制备由纯水、尿素、溶菌酶和牛血清白蛋白构成的模拟液,用此模拟液来代替患者血液,对本文研制的聚醚砜中空纤维膜透析器的透析性能进行评价。

The minor core protein s C-encoding gene of Muscovy duck reovirus was cloned into theprokaryotic expression vector pET32a. The recombinant plasmid pET32a-s C was amplified andextracted after being transformed into E.coli DH5a competent cells. Restriction analysis withEcoRⅠand SacⅠand sequences analysis indicated that the recombinant plasmid was inserted withcorrect open reading frame. The fusion protein about 50 ku was produced after induction with 0.15mmol/L IPTG of E.coli competent cells transformed with pET32a-sC. The SDS-PAGE andWestern-bloting test indicated that the fusion protein reacted with the convalescents sera of duckinfected with Muscovy duck reovirus. The indirect ELISA method was developed by using thepurified fusion s C protein as coating antigen. The optimal concentration of s C was 5μg/ml, thedilution of serum sample was 1:40; The results showed that preparation of an ELISA by using sCas coating antigen in detecting 50 field duck sera in comparison with the AGIP were more sensitiveand specific than agar gel immuno-diffusion AGIP test. The results suggest that presence ofantibody against viral protein sC in duck may be a good indicator by the sC-ELISA for detectionof duck infection with reovirus.

同时,本研究将编码外壳蛋白σC的基因克隆于原核表达载体pET32a上,经过EcoRⅠ和SacⅠ双酶切鉴定和序列分析后,得到阳性重组质粒pET32a-σC;将阳性重组质粒pET32a-σC转化到大肠杆菌BL-21感受态细胞中进行诱导表达,经SDS-PAGE和Western-blbtting检测分析,融合表达的蛋白能够与番鸭呼肠孤病毒感染的康复鸭血清发生特异性反应;将融合表达的蛋白纯化后作为包被抗原,建立了检测鸭血清中呼肠孤病毒抗体的间接酶联免疫吸附试验检测方法,此方法中抗原的最佳包被浓度为5μg/ml、标准阳性血清的最适稀释倍数为1:40倍,用此方法对50份鸭血清样品进行检测,并与琼脂糖凝胶扩散试验检测抗体的法相比较,证明此ELISA方法具有良好的特异性和敏感性,本研究为今后鸭呼肠孤病毒诊断试剂盒的研制奠定了基础。

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