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间接的途径

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We used Aeromonas hydrophila which we separated from the diseased Paralichthys olivaceus to made a inactivated whole cell bacterin .We injected this vaccine in the mice and the Paralichthys olivaceus .Both the mice and the Paralichthys olivaceus gained a good protection of 80% and 70%.The fish with dipping immersion gained a good protection of 85%,and the incidence of disease was 6%.We detected the antibody level in the antiserum by agglutination reaction and indirect ELISA which seemed to be immunogenic crucially.We anatomized the mice then we found that all the purtenance of the immunzied mice are healty.All of this proved that the vaccine is effective.

本实验将所分离到的嗜水气单胞菌用终浓度为0.2%~0.5%的甲醛对菌体进行灭活,制备了灭活疫苗,并优化灭活疫苗的灭活条件;将制备的灭活疫苗分别注射小鼠和牙鲆,均取得了较为理想的效果,其中小鼠的免疫保护率达到80%,牙鲆的免疫保护率达到70%;采用渗透压差法大规模浸泡免疫牙鲆也取得了很好的效果,浸泡免疫过的牙鲆发病率仅为6%,攻毒后免疫保护率为85%;通过应用间接ELISA方法检测受免动物的抗血清,并将牙鲆的抗血清注射新西兰白兔,从而获得兔抗鱼血清,更便于检测,结果都表明动物体内产生了较高的抗体水平;剖检攻毒后的动物,发现免疫过的动物内脏健康无异样,表明该疫苗安全有效,从而为防治牙鲆腹水症开辟了一条新途径,也为进一步制备其基因工程疫苗奠定了基础。

DCs acquired by our reformed methods express both CD83 and CD 14 molecules highly, and have a higher density than other domestic reports. The higher TNF in DCs culture medium of HC patient suggests DCs in patient still have antigen presenting ability and by optimization the culture medium would improve its presenting ability and have a potential value in design and application individual vaccine. Although antigens pulsed DCs have a decrescent antigen presenting ability but BCG HSP70 could induce its mature and improve its presenting ability. Suggests BCG HSP70 would be a useful mature inducer. Lymphocytes primed by DCs based HC vaccine have the specific cytotoxicity against HCC lines. The CTL after freezing and anabiotic could prophylaxis and therapy HC xenograft on nude mouse. The results also suggests that CD4〓 lymphocytes play a important role in HC with a good differentiation and would be useful in treatment this kind of HC. After being activated by Peptide LLNQHACAV of hAFP and apoptotic HCCs pulsed DCs respectively, the culture medium of activated lymphocytes both contains a high level Th1 cytokines IL-12 and TNF. Primed lymphocytes appeared a characteristic of NK cells. DCs not only inhibited the growth of human HCC and other cancer cells in vitro but also prevented the growth of HC xenograft on nude mouse in vivo. There are at least three kinds of mechanism playing important role in DC based vaccine ,there are inhibition of DCs, HC specific CTL and cytokines pathway.

诱导出的DC共同表达CD83和CD14分子,CD83分子表达明显高于国内报道;肝癌患者DC培养上清中TNF水平高于健康人,提示肝癌患者DC仍具一定的抗原呈递能力,适当调控可使其行使APC功能,以期在肝癌个体化疫苗中发挥作用;DC负载肝癌可溶性抗原后,抗原呈递能力降低,BCG HSP70可促进DC成熟,增加其抗原呈递能力,预示BCG HSP70有可能成为促进DC活化和成熟的另一重要分子;肝癌DC疫苗在体外诱导肝癌特异性淋巴细胞,活化的淋巴细胞在体外对肝癌细胞的杀伤以特异性CTL为主,同时分泌较高水平Th1型细胞因子IL-12和TNF,并抑制4种肝癌细胞生长;冷冻复苏后的肝癌特异性淋巴细胞可以预防和抑制人肝癌裸鼠皮下移植瘤,提示DC负载肝癌可溶性抗原后诱发的MHC-Ⅱ类限制性CD4〓T细胞有可能在分化程度高的肝癌治疗中发挥作用;用DC和HLA-A2〓DC分别负载凋亡肝癌细胞和hAFP218-226位LLNQHACAV HLA-A2限制性九肽,在体外诱导肝癌特异性淋巴细胞,活化后的CTL细胞分泌较高水平的Th1型细胞因子IL-12和TNF,并具较强杀伤活性,此CTL同时具备NK细胞特征;DC对肿瘤细胞的抑制作用可能是通过吞噬实现的,Fas-L在DC抑制中也起一定作用;DC对人肝癌裸鼠皮下移植瘤的抑制率为97%;在肝癌DC疫苗的作用中,至少联合3种以上机制,即通过DC的直接作用、肝癌特异性CTL和细胞因子途径直接或间接地杀伤和抑制肝癌细胞。

That means Pho85 kinaseand calcineurin were involved in salt tolerance with an identical target protein or in 中科院上海生化与细胞所博士学位论文摘要 the same pathway. Inhibition of calcineurin decrease the YPH499, pho80? mutant,and pap1 (pcl7)? mutant Mn2+ tolerance but not that of pho85? mutant and thepho85? mutant was more sensitive to Mn2+ than YPH499, pho80? mutant, and pap1(pcl7)? mutant even with the addition of cyclosporin A. Therefore, the conclusioncould be drawn that PHO85 gene played a dominant role in Mn2+ homeostasisregulation in compare with calcineurin. As for Ca2+ tolerance, cyclosporin A canincrease the tolerance to Ca2+ of all the mutant mention above, that means Pho85kinase and calcineurin function antagonistically in regulation of Ca2+ homeostasis.In Bioinformatics, BRI3 gene is a novel gene without any function clue but givehigh conservation in mammalian.

我们通过钙调磷酸酶的特异抑制剂环孢菌素A研究了YPH499、+2+pho85缺失株、pho80缺失株、pap1缺失株在钙调磷酸酶失活后对Na、Mn、2+Ca金属离子敏感性的变化,结果显示Pho85蛋白激酶和钙调磷酸酶通过直接+或间接激活同一个靶蛋白或途径来增强细胞对Na的耐受;和钙调磷酸酶相比,2+PHO85的缺失对酵母细胞Mn耐受性的破坏是相对控制性的,钙调磷酸酶的失2+活不能进一步降低pho85缺失株对Mn的耐受能力;Pho85和钙调磷酸酶在对2+Ca的耐受调节中是相互拮抗的,钙调磷酸酶的失活能增加pho85缺失株和2+pho80缺失株的Ca耐受。i中科院上海生化与细胞所博士学位论文摘要对本实验室克隆的人新基因BRI3进行系统的生物信息学分析,发现它是一个在哺乳动物中保守的,但功能未知的基因。

One was that one molecule of nitrous acid di-rectly reacted with DMA,and the other was that two molecules of nitrous acid reacted first to yield nitrogen trioxide,which then reacted with DMA.

计算结果表明,DMA与亚硝酸间接反应的活化能比直接反应的低约55kJ/mol,由此可推断DMA与亚硝酸生成NDMA的反应是以两分子的亚硝酸先生成ONNO2再亚硝化DMA为主要反应途径。

As to direct correl...

文中评述了间接对比方法的各个步骤的要点及直接对比方法的可能途径。

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