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Methods:(1) ADM was produced from swine skins treated with trypsin followed by Triton X-100.(2) type I collagen of mouse tail was extracted by 0.5mol/L acetoacetic acid.(3) Two kinds of ADM were got by being soaked with 0.02% glutaraldehyde for 5~10 min or being soaked with mouse type I collagen for 24h, then preserved at 4℃.

(1)应用胰蛋白酶消化-去污剂法制备ADM；(2)用0.5mol/L乙酸溶液提取制备I型鼠尾胶原；(3)用0.02%戊二醛溶液浸泡ADM5-10min，制成交联型ADM；另一部分ADM用质量分数0.25%的I型鼠尾胶原浸泡24h，制成胶原包埋型ADM，冰冻保存；(4)用酶消化法培养原代成纤维细胞，取第三代增殖期细胞，将其调整至浓度为2×105/ml的成纤维细胞悬液。

"PP Mouse" is nickname from her husband, as she was born in the last month of Chinese mouse year, so he leave this agname for her, regarding it is such lovely agname, so she took it delightly.

皮皮鼠是老公给她起的昵称，因为她是鼠年腊月生的，抓住了小老鼠的一点小尾巴，所以老公就给也这么一个绰号，挺可爱的名字，她也乐的接受了。

At the beginning the number was decrease rapidly, then slowly. There were still some MSCs alive 8 week after transplant. Conclusion: The mouse's MSCs can survive more than 8 week in subcutaneous tissue of allogene mouse. MSCs were hypoimmunogenic.

小鼠MSCs植入同种异体小鼠皮下8周后仍能存活，MSCs在体内保持低免疫原性的特点，同种异体与同种同基因MSCs在诱导机体免疫应答方面无明显差异。

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。

Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC；分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态；流式细胞术检测DC表型；HRP吞噬实验测定DC的群体内吞能力；MTT法检测同种异体混合淋巴细胞反应；ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平；冻融法制备肝癌细胞可溶性抗原；流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性；MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响；SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度，ELISA测定活性；流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型；MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应；流式细胞术检测肝癌细胞表面HLA-DR表达；MTT法检测肝癌DC疫苗对自身淋巴细胞的活化；原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达；流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L；MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用；Cell-ELISA检测人肝癌细胞hAFP表达；MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响；ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平；肝癌细胞凋亡的诱导和检测；DC吞噬凋亡肝癌细胞后的电子显微镜观察；DC对肝癌细胞的生长抑制试验；人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定；DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤；冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

The anucleate but rich in globin mRNA reticulocytes of normal mouse were hybridized with mouse lymphoma cells.

本实验选用富含有血红蛋白特征面无核的正常小鼠网织红细胞，与小鼠淋巴瘤细胞株融合，对杂交细胞基因产物血红蛋白进行联苯胺组织化学染色、荧光抗体和生化测定。

MethodsThe immunosuppressive mouse model was established by using hydrocortisone. Huangqi Fuzhengtang and Yupingfeng San apozema were given to the mice intragastrically. The contents of hemolysin, IL-2, and INF-γ in the mouse serum and the expression of CD3+, CD4+, and CD8+ in the peripheral blood lymphocytes were measured.

以氢化可的松造成免疫低下小鼠模型，用黄芪扶正汤和玉屏风散水煮液进行灌胃，检测小鼠血清溶血素的生成、IL-2和INF-γ的含量以及外周血淋巴细胞CD3+，CD4+和CD8+的表达。

The genetic relationship between pig and bovine who are both Artiodactylous is the nearest, the next is human, and the farthest is mouse. The differentiation sequence taken place in four species from the same ancestor is that mouse is the earliest one, and the latter human, and pig and bovine are the latest.

种系发生分析结果表明，作为偶蹄目的猪和牛的遗传关系最近，其次是人类，小鼠与家猪和牛的遗传关系最远。4种动物从共同祖先分化的顺序分别为小鼠最早，人类次之，然后为猪和牛。

In the cytosol preparations, nimodipine inhibited the iNOS enzymatic activity induced by LPS +〓 in mouse astroglia cells with an 〓 value of 0.036±0.003mM and nNOS activity in mouse neurons with an 〓 value of 0.047±0.003mM, but did not affect the eNOS activity in bovine cerebral endothelial cells.

在体外培养细胞制备的匀浆液，尼莫地平抑制由LPS+〓诱导的星型胶质细胞的iNOS活性，〓为0.036±0.003mM，同时还抑制神经元的nNOS活性，IC50为0.047±0.003mM，但对牛脑微血管内皮细胞的eNOS活性无明显影响。

Barometz was significantly effective on the mouse with AA and mouse with DK-AA. And the sample processed with sand was obviously stronger than the raw one.

实验表明，狗脊对肾阳虚佐剂性关节炎大鼠及佐剂性关节炎大鼠具有治疗作用，炮制后作用增强。

For a big chunk of credit-card losses; the number of filings (and thus charge-off rates) would be rising again, whether

年美国个人破产法的一个改动使得破产登记急速下降，而后引起了信用卡大规模的亏损。

Eph. 4:23 And that you be renewed in the spirit of your mind

弗四23 而在你们心思的灵里得以更新