性原细胞

Results:(1)NSCs form typical neurospheres under adequate concentration in vitro, which are immunoreactive to Vimentin. Typically and terminally differentiated mature neural cells could not be found without the stimulus of mitogen or only under NSCs self-regulation and self-induction;(2)NSCs derived from hippocampus maintain the character of stem cells much longer with better biological behavior; NSCs passed to the 2-3 passage are the best to graft since they have not differentiated;(3)NSCs cultured in vitro could self-regulate and differentiate into neurospheres and progenitors positively immunoreactive to specific antibodies representing neurons, astrocytes, oligodendrocytes and Schwann cells;(4)There are widespread synaptic contacts between various kinds of descendent clones and cells;(5)Neurospheres could be formed without the stimulus of mitogen when NSCs and OECs are cocultured. Many neurospheres and cells immunoreactive to Vimentin, GFAP, MAP2, 02, p75NGFR, GFAP, S-100, Synaptosis, Vimentin, Tau (Tau is only positive in cocultureof HNSCs+HOECs) could be found;(6)The supernatant fluid triturated from adult rat spinal cord stimulates NSCs to differentiate into neurons, but do not terminally differentiate;(7)Fibroblasts and O4 oligodendrocytes are not supported to grow under this culture medium.Part II: Isolation, culture and identification of rat and human olfactory ensheathing cellsOlfactory ensheathing cells/glials are the most powerful cells to enable the regeneration of axons in the central nervous system.

结果表明：①在适宜的浓度体外培养条件下，NSCs能形成典型的神经干细胞克隆球，Vimentin免疫荧光染色阳性，单靠丝裂原刺激或NSCs自我调节和分化诱导，不会产生典型的终末分化的成熟神经细胞；②海马源性的NSCs维持干细胞特性的时间更长，生物学特性更优；③传至第2～3代的NSCs尚未分化时移植最佳；④体外培养的NSCs能自我调控分化为神经元、星形胶质细胞、O2少突胶质细胞、雪旺氏细胞染色阳性克隆球和前体细胞；⑤各种子代克隆球和细胞存在广泛的突触联系；⑥NSCs与OECs联合培养时，不需丝裂原刺激即能形成克隆球，获得大量Vimentin、GFAP、MAP2、O2、p75NGFR、GFAP、S-100、Synaptosis、Vimentin、Tau(Tau只有人HNSCs+HOECs联合培养时出现阳染)染色阳性的克隆球和细胞；⑦脊髓研磨后的上清液刺激神经干细胞向神经元方向分化，但并不出现终末分化；⑧本研究培养条件不利于成纤维细胞、O4生长。

Based on these previous work, in this research, in order to explore the the molecular mechanism of enhanced tumor cell immunogenicity treated by elemene and mytomycin C compositely, we used murine Hca-F ascitic hepatic carcinoma ( high lymphatic metastasis substrain of H22) as an experimental model to compare the immunoprotective effects on Hca-F cells as well as the mechanism of the effects caused by HSP70-peptide complex purified from untreated tumor cells, EC-TCV and BCG; We studied the mechanism of increased tumor cell membrane HSP expression treated with elemene by the techniques of RT-PCR and immunoelectron microscopy; In order to deeply elucidate the antitumor effects and the molecular mechanism of elemene in increasing tumor cell immunogenicity we analyzed and compared the effects of elemene and heat shock on the gene expressions of tumor cell using gene expression profile chip.

本论文在上述研究成果的基础上，选择小鼠肝癌H22的高淋巴道转移亚系Hca-F为模型，进一步比较了未处理肿瘤细胞来源、榄香烯复合瘤苗来源的和卡介苗来源的HSP70-肽复合物对Hca-F的免疫保护效应、效应机制，探讨榄香烯-丝裂霉素复合处理增强肿瘤细胞免疫原性的分子机制；以Hca-F和人肝癌细胞株HepG〓等为模型研究了榄香烯处理增强肿瘤细胞膜热休克蛋白表达的机制；以HepG〓为模型用表达谱基因芯片研究比较了榄香烯与热休克处理对肿瘤细胞基因表达的影响，从而进一步阐明榄香烯抗肿瘤作用及增强肿瘤细胞免疫原性的分子机制。

BioActive Glycopeptides and Peptidoglycans promote bradykinin action and activate glomerulus and nephrocyte to make vasopressor factor (angiotension-I)-suppressing as ACE-blocking to anti-hypertension and against Chronic fatigue syndrome, Nephrosis, glomerulonephritis, congestive heart failure.

天然生物活性醣胜肽与肽聚醣，可作为医师处方调配最好的医用营养素补充剂：提升血管舒缓激肽的活动、促进活化肾小球与肾原细胞，抑制血管增压因子、高血压、对抗慢性疲劳综合症、肾变病、血管球性肾炎、充血性心脏衰竭。

BioActive Glycopeptides and Peptidoglycans promote bradykinin action and activate glomerulus and nephrocyte to make vasopressor factor (angiotension-I)-suppressing as ACE-blocking to anti-hypertension and against Chronic fatigue syndrome, Nephrosis, glomerulonephritis, congestive heart failure.

天然生物活性醣胜肽与肽聚醣，可作为医师处方调配最好的医用营养素补充剂：提升血管舒缓激肽的活动、促进活化肾小球与肾原细胞，抑制血管增压因数、高血压、对抗慢性疲劳综合症、肾变病、血管球性肾炎、充血性心脏衰竭。

①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法：(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC，细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构；(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定；(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC，并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况，根据各个细胞克隆受Dox调控表达的程度，选择低背景、高诱导表达AT2R的细胞系，作为双重稳定MSC细胞系；蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况；(4)建立大鼠颈动脉球囊损伤动物模型，将双重稳定MSC在术中导入血管，分别于14 d、28 d进行病理切片，检测可调控AT2R对新生内膜增生的影响；采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变，TUNEL法检测血管组织中细胞凋亡的变化情况。

Apoptotic cells have immunogenicity,and may be the major source of autoantigens in lupus BXSB mice.Lupus genetic factors and Yaa gene can potentiate the immunogenicity of apoptotic cells or accelerate their inducing the lupus nephritis.

凋亡细胞具有免疫原性，它可能是狼疮性BXSB小鼠体内自身抗原的主要来源；狼疮性遗传因素和Yaa基因能促进凋亡细胞的免疫原性和增强凋亡细胞的致肾炎作用。

Apoptotic cells have immunogenicity,and may be the major source of autoantigens in lupus BXSB mice.Lupus gecnic factors and Yaa gene can potentiate the immunogenicity of apoptotic cells or accelerate their inducing the lupus nephritis.

凋亡细胞具有免疫原性，它可能是狼疮性BXSB小鼠体内自身抗原的主要来源；狼疮性遗传因素和Yaa基因能促进凋亡细胞的免疫原性和增强凋亡细胞的致肾炎作用。

ConclusionApoptotic cells have immunogenicity,and may be the major source of autoantigens in lupus BXSB mice.Lupus genetic factors and Yaa gene can potentiate the immunogenicity of apoptotic cells or accelerate their inducing the lupus nephritis.

结论凋亡细胞具有免疫原性，它可能是狼疮性BXSB小鼠体内自身抗原的主要来源；狼疮性遗传因素和Yaa基因能促进凋亡细胞的免疫原性和增强凋亡细胞的致肾炎作用。

Apoptotic cells have immunogenicity,and may be the major source of autoantigens in lupus BXSB mice.Lupus genetic factors and Yaa gene can potentiate the immunogenicity of apoptotic cells or accelerate their inducing the lupus nephritis. Apoptosis; Mice,BXSB; Lupus erythematosus,systemic

凋亡细胞具有免疫原性，它可能是狼疮性BXSB小鼠体内自身抗原的主要来源；狼疮性遗传因素和Yaa基因能促进凋亡细胞的免疫原性和增强凋亡细胞的致肾炎作用。

Apoptotic cells have immunogenicity,and may be the major source of autoantigens in lupus BXSB mice.Lupus genetic factors and Yaa gene can potentiate the immunogenicity of apoptotic cells or accelerate their inducing the lupus nephritis. Apoptosis; Mice,BXSB; Lupus erythematosus,systemic

凋亡细胞常识具有免疫病毒原性，它可能流产是狼疮性BXSB小鼠体内自身抗原的主要来源；狼疮性遗传因素和Yaa基因能促进凋亡细胞常识的免疫病毒原性和增强凋亡细胞常识的致肾炎作用。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐