无细胞的

RESULTS:①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs: The observation result of phase contrast microscopy showed that, endothelial cells planted on the PEG-PLA-PGL membranes began to attach and stretch after being planted 4-6 hours. Three days later, cells grew in colonies rapidly, after 5 days, colonies began to fuse and seemed like cobble-stone. The cells were shuttle or polygon in shape after passages.

结果：①人脐静脉内皮细胞与聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片的细胞相容性：相差显微镜下种植在聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片上的细胞4~6 h后即开始贴壁、伸展，3 d后细胞开始呈集落样生长，生长较快，5 d后细胞集落开始融合，呈现特征性的鹅卵石样形态，经多次传代后，细胞呈长梭形或多角形，与对照组相比无明显差异。

There are three types of hemocytes: agranular hemocyte, large granular hemocyte and small granular hemocyte.

正常中华绒螯蟹的血细胞可分为三类：无颗粒细胞、小颗粒细胞和大颗粒细胞，而RLOs只特异性地侵染小颗粒细胞。

Results: TFF2 peptide was distributed in cytoplasm of epithelial cells of the intercalated duct, striated duct and interlobular duct, and positive reaction was found in the cavity, especially near the cavosurface of epithelial cells in the striated duct and interlobular ducts.

结果：TFF2免疫反应阳性物质主要位于闰管、纹状管和小叶间导管上皮细胞胞质，特别是近管腔面更多，纹状管和小叶间导管管腔内亦有阳性物质分布；颗粒区管的少颗粒暗细胞或无颗粒细胞强阳性；多颗粒细胞弱阳性或阴性。

In treatment group, there was no progressive necrosis in stasis zone, and at 24 h post injury, capillary dilation, cell edema and inflammatory infiltration were lessened significantly. In control group, 2 wounds had progressive necrosis (accounted for 10% of the total). Cell edema, inflammatory infiltration and capillary thrombosis were serious, Until 72 hr post injury, cell edema did not subside and necrosis of the dermis worsened. 2. Pathological assessment of the stasis zone tissue: As compared with that in control group, in treatment group the dermis structure in stasis zone was intact and the collagenous fiber bundle was normal.

结果：①创面细胞形态学改变：伤后8h毛细血管扩张、细胞水肿，炎性浸润最明显；治疗组中央淤滞区无进行性坏死，伤后24h后毛细血管扩张、细胞水肿、炎性浸润减轻；对照组2个创面呈进行性坏死（占10%），中央淤滞区细胞水肿、炎性浸润明显，毛细血管血栓形成，伤后72h水肿仍明显，真皮坏死加重；②淤滞区组织病理学评分：治疗组与对照组相比，治疗侧淤滞区的表皮结构相对完整、胶原纤维束相对正常，粒细胞浸润小于5个/400倍视野。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实：两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用，并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著，最佳效应浓度为400nM；2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响：①CREG蛋白对VSMC迁移的影响：刮伤实验发现，加入最佳效应浓度的wtCREG和mCREG蛋白24h后，OB2组迁移能力下降，HITASY组无明显变化；细胞外基质金属蛋白酶-2，9(Matrix metallo-proteinase 2，9，MMP2 ，9)明胶酶电泳检测和Western blot检测结果证实，两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2，9减少，而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases，TIMPs)增加；②CREG蛋白对VSMC分化的影响：加入400nM的wtCREG和mCREG蛋白12h后，OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加，LM-1、FN表达减少；③流式细胞仪分析细胞周期和BrDU染色分析证实，加入400nM的wtCREG和mCREG蛋白后，OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572，HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034；3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用：①免疫共沉淀和免疫荧光双染色分析结果显示，CREG蛋白与M6P/IGF2R存在直接结合；②应用抗体阻断实验：将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中，CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱，而且与加入anti- M6P/IGF2R浓度正相关。

The expression of nestin antigen, NSE and GFAP were identified by immunohistochemistry, and the number of NSE-positive cells and GFAP-positive cells were dividedly counted.

结果大鼠胚胎神经管神经上皮干细胞在无血清培养基中可形成大量呈nestin抗原阳性细胞构成的神经球，经传代有血清培养后分化为NSE阳性和GFAP阳性细胞，其中NSE阳性细胞占细胞总数的47.7％，GFAP阳性细胞占细胞总数的39.8％。

Affect on the liver pathological state after XIEZHIPING treated: the golden hamster hepatic tissue morphous structure of normal control was normal, which had regular hepatic lobules and no lipid droplet;that of model control manifested diffuse heavy adipose degeneration and heavy ballooning degeneration with inflammatory cell infiltrate which mainly composed of lympholeukocyte;that of high dose group showed midrange ballooning degeneration with chronic inflammatory cell infiltrate which mainly composed of lympholeukocyte and plasmocyte;that of media dose group indicated slight focus ballooning degeneration with a small quantity normal hepatic cells in some locals;that of low dose group appeared midrange adipose degeneration with a few lympholeukocyte cell infiltrate in the parts of central veins and port area.

泻脂平治疗后对肝脏病理状况的影响：空白对照组金黄地鼠的肝组织形态结构正常，肝小叶规则，无脂滴发现；模型对照组肝脏弥漫性重度脂肪变性，发生重度气球样变，伴见以淋巴细胞为主的慢性炎性细胞浸润；泻脂平高剂量组肝细胞呈现中度气球样变，伴见以淋巴细胞、浆细胞为主的慢性炎性细胞浸润；中剂量组肝细胞轻度灶性气球样变，局部可见少量正常肝细胞；低剂量组肝细胞呈中度脂肪变性，可见中央静脉及汇管区些许淋巴细胞浸润。

Affect on the liver pathological state afterXIEZHIPING treated: the golden hamster hepatic tissue morphous structureof normal control was normal, which had regular hepatic lobules and nolipid droplet: that of model control manifested diffuse heavy adiposedegeneration and heavy ballooning degeneration with inflammatory cell infiltrate which mainly composed of lympholeukocyte; that of high dosegroup showed midrange ballooning degeneration with chronic inflammatorycell infiltrate which mainly composed of lympholeukocyte and plasmocyte;that of media dose group indicated slight focus ballooning degenerationwith a small quantity normal hepatic cells in some locals; that of lowdose group appeared midrange adipose degeneration with a fewlympholeukocyte cell infiltrate in the parts of central veins and portarea.

泻脂平治疗后对肝脏病理状况的影响：空白对照组金黄地鼠的肝组织形态结构正常，肝小叶规则，无脂滴发现；模型对照组肝脏弥漫性重度脂肪变性，发生重度气球样变，伴见以淋巴细胞为主的慢性炎性细胞浸润；泻脂平高剂量组肝细胞呈现中度气球样变，伴见以淋巴细胞、浆细胞为主的慢性炎性细胞浸润；中剂量组肝细胞轻度灶性气球样变，局部可见少量正常肝细胞；低剂量组肝细胞呈中度脂肪变性，可见中央静脉及汇管区些许淋巴细胞浸润。

The hFCECs were cultured by sticking tissues piece method that the cornea were divided into the limbus and central tissue piece and digestive method,respectively.For digestive method,the digestive juice of 5mg/ml DispaseⅡ+0.25%trypsin was chosed.D/F12 with 10%fetal bovine serum and 100 U/ml penicillin and streptomycin were used in this study,the culture condition was 37℃and 5%CO2,and culture medium changed every three days2、Passage of hFCECsAfter more than 80%confluenced,cells from corneal limbus hFCECs were digested for 5-15min with different concentrations of trypsin/EDTA at 37℃,and then passaged at a ratio of 1:2.The cells were subcultured on empty plates,mouse 3T3 fibroblast feeder layer,fetal corneal stromal cell feeder l ayer or HTK feeder layer respectively.And D/F12, mouse 3T3 fibroblast conditioned medium,fetal corneal stromal cells conditioned medium or HTK conditioned medium with 10%FBS and 100 U/ml penicillin and streptomycin were used respectively as the culture medium.And the culture medium were changed every three days as well.

方法一、人胎儿角膜上皮原代和传代培养1、原代培养严格无菌操作获取人胎儿角膜片，采用组织块贴壁法、酶消化法原代培养人胎儿角膜上皮细胞。2、传代培养角膜缘部的细胞生长融合达80%以上，不同稀释浓度的胰酶/EDTA消化液消化传代分别接种于空板、小鼠3T3成纤维细胞饲养层、胎儿角膜基质细胞饲养层及HTK饲养层，培养液分别采用D/F12、小鼠3T3成纤维细胞条件培养液、胎儿角膜基质细胞条件培养液、HTK条件培养液。

The Plant tissue raise is the plant sterile culture technology, has the totipotency theory according to the vegetable cell, the use plant body exsomatize organ (for example root, stem, leaf, stem point, flower, fruit and so on), the organization (for example cambium, epidermis, cerebral cortex, marrow department cell, endosperm and so on) or the cell (for example big spore, small spore, somatic cell and so on) as well as the protoplast, in aseptic and suitable artificial culture medium and illumination, temperature and so on under artificial condition, can induce injuries the organization, not to decide the bud, the adventitious root, finally forms the complete adult plant the discipline.

植物组织培养即植物无菌培养技术，是根据植物细胞具有全能性的理论，利用植物体离体的器官(如根、茎、叶、茎尖、花、果实等)、组织（如形成层、表皮、皮层、髓部细胞、胚乳等）或细胞（如大孢子、小孢子、体细胞等）以及原生质体，在无菌和适宜的人工培养基及光照、温度等人工条件下，能诱导出愈伤组织、不定芽、不定根，最后形成完整的植株的学科。

The labia have now been sutured together almost completely.The drains and the Foley catheter come out at the top.

此刻阴唇已经几乎完全的缝在一起了，排除多余淤血体液的管子和Foley导管从顶端冒出来。

To get the business done, I suggest we split the difference in price.

为了做成这笔生意，我建议我们在价格上大家各让一半。