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1.
INTRODUCTION Cell encapsulation in biocompatible and semipermeable polymeric membranes is an effective method for immunoprotection, regardless of the type of recipient (allograft, xenograft, etc.). The semipermeable nature of the membrane prevents high molecular weight molecules, antibodies and other immunologic moieties from coming into contact with the encapsulated cells and destroying them as foreign invaders, but permits the entry of nutrients and oxy- gen and the exit of therapeutic p… 相似文献
2.
Peng-yan QIAO Fang-fang LI Li-min DONG Tao XU Qiu-fei XIE 《Journal of Zhejiang University. Science. B》2014,15(4):382-392
研究目的：本研究应用海藻酸钠-壳聚糖微囊保护成骨细胞,接种到β-磷酸三钙/磷酸钙骨水泥（β-TCP/CPC）浆料中,使β-TCP/CPC骨修复材料具有一定的细胞活性,同时提高固化后材料的孔隙率和孔径,以最终实现提高β-TCP/CPC骨水泥的降解速度,加快成骨和骨修复。创新要点：本研究首次应用海藻酸钠-壳聚糖微胶囊包封成骨细胞与CPC浆料复合,复合后实现自动细胞释放,释放出的细胞具有良好的生物学活性。研究方法：（1）高压静电成囊法制备载小鼠成骨前体细胞（MC3T3-E1）的海藻酸钙和海藻酸钠-壳聚糖微胶囊;（2）微囊化MC3T3-E1细胞,进行体外培养,使用细胞计数试剂盒（CCK-8）检测细胞活性,并用钙黄绿素-AM（Calcein-AM）和碘化丙啶（PI）进行活死细胞双重染色;（3）微囊化MC3T3-E1细胞与β-TCP/CPC浆料复合培养后,激光共聚焦扫描显微镜和环境扫描电子显微镜观测细胞在材料上的释放、粘附,CCK-8法检测材料上细胞的活力,碱性磷酸酶（ALP）检测观察细胞的分化状况,茜素红染色观察释放细胞的矿化能力。重要结论：海藻酸钠-壳聚糖微胶囊可作为可注射磷酸钙骨水泥内部接种成骨细胞并实现细胞释放的良好载体,释放出的成骨细胞具有良好的生物学活性。 相似文献
3.
Zhan-mei Jiang Li-na Bai Nan Yang Zhi-biao Feng Bo Tian 《Journal of Zhejiang University. Science. B》2017,18(10):867-877
The stability of β-carotene microcapsules using Maillard reaction products (MRPs) derived from whey protein isolate (WPI) and galactose as coating materials, was studied under the varying environmental conditions of temperature, pH, air, incandescent light, and ultraviolet (UV) light. Scanning electron microscopy showed that microcapsules prepared by WPI-galactose MRPs displayed a smooth and less concave-convex surface and that the particle size (D50) of the microcapsules made with WPI-galactose MRPs was smaller than those made with WPI-galactose mixture. The storage stability of β-carotene microencapsulated in WPI-galactose MRPs was remarkably better than that of β-carotene microencapsulated in the WPI-galactose mixture and that of β-carotene crystal, in respect of temperature, pH, air, incandescent light, and UV light measurements. When the storage temperature was increased from 5 to 105 °C, the retention rate of β-carotene microcapsules significantly decreased (P<0.05). When pH values were increased from 1 to 12, the β-carotene retention rate of the microcapsules significantly increased and afterward decreased. Compared with the retention rate of β-carotene microencapsulated in a WPI-galactose mixture, the retention rate of β-carotene microencapsulated in WPI-galactose MRPs was at a maximum between pH 8 and 9. Under the actions of air, incandescent light, and UV light, the retention rates of β-carotene microcapsules in WPI-galactose MRPs and WPI-galactose mixture, as well as in β-carotene crystal, decreased significantly as the storage time increased (P<0.05). Therefore, the use of WPI-galactose MRPs as coating materials can aid in improving the storage stability of β-carotene microcapsules. 相似文献
4.
Zhao Q Liu ZD Xue Y Wang JF Li H Tang QJ Wang YM Dong P Xue CH 《Journal of Zhejiang University. Science. B》2011,12(7):534-544
Ds-echinoside A (DSEA), a non-sulfated triterpene glycoside, was isolated from the sea cucumber Pearsonothuria graeffei. In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion, migration, invasion, and
angiogenesis. In this study, we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells
Hep G2, with a half-maximal inhibitory concentration (IC50) of 2.65 μmol/L, and suppressed Hep G2 cell adhesion, migration, and invasion in a dose-dependent manner. DSEA also reduced
tube formation of human endothelial cells ECV-304 on matrigel in vitro and attenuated neovascularization in the chick embryo
chorioallantoic membrane (CAM) assay in vivo. Immunocytochemical analysis revealed that DSEA significantly decreased the expression
of matrix metalloproteinase-9 (MMP-9), which plays an important role in the degradation of basement membrane in tumor metastasis
and angiogenesis. DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase-1 (TIMP-1), an
important regulator of MMP-9 activation. From the results of Western blotting, the expressions of nuclear factor-kappa B (NF-κB)
and vascular endothelial growth factor (VEGF) were found to be remarkably reduced by DSEA. These findings suggest that DSEA
exhibits a significant antimetastatic activity through the specific inhibition of NF-κB-dependent MMP-9 and VEGF expressions. 相似文献
5.
Ya-fei Shi Ju-fang Chi Wei-liang Tang Fu-kang Xu Long-bin Liu Zheng Ji Hai-tao Lv Hang-yuan Guo 《Journal of Zhejiang University. Science. B》2013,14(8):696-704
Objective: To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 (MMP-2) and tissue inhibitors of matrix metalloproteinase-2 (TIMP-2) and on cell migration of cultured rat vascular smooth muscle cells (VSMCs). Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Methods: Rat VSMCs were incubated with different concentrations of homocysteine (50–5 000 μmol/L). Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin (10−9–10−5 mol/L) were added when VSMCs were induced with 1 000 μmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Results: Homocysteine (50–1 000 μmol/L) increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5 000 μmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and activation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine (50–5 000 μmol/L) stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Conclusions: Homocysteine (50–1 000 μmol/L) significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression and activation of MMP-2 and abnormal migration of VSMCs induced by homocysteine. 相似文献
6.
Objective:To investigate the effects of mycotoxin moniliformin (MON) on the metabolism of aggrecan and type II collagen in human chondrocytes in vitro and the relationship between MON and Kashin-Beck disease (KBD).Methods:Human chondrocytes were isolated and cultured on bone matrix gelatin to form an artificial cartilage model in vitro with or without MON toxin.Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.The expression of aggrecan and type II col... 相似文献
7.
Xu Q Li ZX Peng HQ Sun ZW Cheng RL Ye ZM Li WX 《Journal of Zhejiang University. Science. B》2011,12(4):247-255
This paper aims to investigate the effects of artesunate (ART) on growth and apoptosis in human osteosarcoma HOS cell line
in vitro and in vivo and to explore the possible underlying mechanisms. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT) assay. The induction of apoptosis was detected by light and transmission electron microscopy and flow cytometry.
Western blot analysis was used to investigate the related mechanisms. Nude mice were further employed to investigate the antitumour
activity of ART in vivo. MTT assay results demonstrated that ART selectively inhibits the growth of HOS cells in a dose- and
time-dependent manner. Based on the findings of light and transmission electron microscopy, Hoechst 33258 staining, and fluorescein
isothiocyanate (FITC)-annexin V staining, the cytotoxicity of ART in HOS cells occurs through apoptosis. With ART treatment,
cytosolic cytochrome c was increased, Bax expression was gradually upregulated, Bcl-2 expression was downregulated, and caspase-9 and caspase-3
were activated. Thus, the intrinsic apoptotic pathway may be involved in ART-induced apoptosis. Cell cycle analysis by flow
cytometry indicated that ART may induce cell cycle arrest at G2/M phase. In nude mice bearing HOS xenograft tumours, ART inhibited tumour growth and regulated the expressions of cleaved
caspase-3 and survivin, in agreement with in vitro observations. ART has a selective antitumour activity against human osteosarcoma
HOS cells, which may be related to its effects on induction of apoptosis via the intrinsic pathway. The results suggest that
ART is a promising candidate for the treatment of osteosarcoma. 相似文献
8.
Edaravone protects PC12 cells from ischemic-like injury via attenuating the damage to mitochondria 总被引：6，自引：0，他引：6
Background： Edaravone had been validated to effectively protect against ischemic injuries. In this study, we investigated the protective effect of edaravone by observing the effects on anti-apoptosis, regulation of Bcl-2/Bax protein expression and recovering from damage to mitochondria after OGD （oxygen-glucose deprivation）-reperfusion. Methods： Viability of PC 12 cells which were injured at different time of OGD injury, was quantified by measuring MTT （2-（4,5-dimethylthia-zol-2-yl）-2,5-diphenyltetrazolium bromide） staining. In addition, PC 12 cells＇ viability was also quantified after their preincubation in different concentration of edaravone for 30 min followed by （OGD）. Furthermore, apoptotic population of PC 12 cells that reinsulted from OGD-reperfusion with or without preincubation with edaravone was determined by flow cytometer analysis, electron microscope and Hoechst/Pl staining. Finally, change of Bcl-2/Bax protein expression was detected by Western blot. Results： （1） The viability of PC12 cells decreased with time （1-12 h） after OGD. We regarded the model of OGD 2 h, then replacing DMEM （Dulbecco＇s Modified Eagle＇s Medium） for another 24 h as an OGD-reperfusion in this research. Furthermore, most PC 12 cells were in the state of apoptosis after OGD-reperfusion. （2） The viability of PC 12 cells preincubated with edaravone at high concentrations （1, 0.1, 0.01 μmol/L） increased significantly with edaravone protecting PC 12 cells from apoptosis after OGD-reperfusion injury. （3） Furthermore, edaravone attenuates the damage of OGD-reperfusion on mitochondria and regulated Bcl-2/Bax protein imbalance expression after OGD-reperfusion. Conclusion： Neuroprotective effects of edaravone on ischemic or other brain injuries may be partly mediated through inhibition of Bcl-2/Bax apoptotic pathways by recovering from the damage of mitochondria. 相似文献
9.
Zhi Yang Miao Liu Yin-gang Zhang Xiong Guo Peng Xu 《Journal of Zhejiang University. Science. B》2009,10(3):188-192
Objective: We investigated the effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells (BMSCs) in vitro. Methods: BMSCs were isolated from adult marrow donated by a hip osteoarthritis patient with prosthetic replacement and cultured in vitro. The third passage cells were divided into negative pressure treatment group and control group. The treatment group was induced by negative pressure intermittently (pressure: 50 kPa, 30 rain/times, and twice daily). The control was cultured in conventional condition. The osteogenesis of BMSCs was examined by phase-contrast mi-croscopy, the determination of alkaline phosphatase (ALP) activities, and the immunohistochemistry of collagen type I. The mRNA expressions of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) in BMSCs were analyzed by real-time poly-merase chain reaction (PCR). Results: BMSCs showed a typical appearance of osteoblast after 2 weeks of induction by intermit-tent negative pressure, the activity of ALP increased significantly, and the expression of collagen type 1 was positive. In the treatment group, the mRNA expression of OPG increased significantly (P<0.05) and the mRNA expression of OPGL decreased significantly (P<0.05) after 2 weeks, compared with the control. Conclusion: Intermittent negative pressure could promote os-teogenesis in human BMSCs in vitro. 相似文献
10.
Objective: To investigate the infection of human embryo fibroblast cell line HF cells by CMV as well as the effects of CMV
on β-actin mRNA and microfilaments. Methods: HF cells shape was observed after the infection of CMV. RT-PCR assay was used
to detect the mRNA expression of CMV immediate early (IE) gene, β-actin and GAPDH genes of HF cells infected by CMV. CMV particles
and cell microfilaments were detected with electron microscope. Results: Shape of HF cell changed after the infection by CMV.
HF cells infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time-and titer-dependent
manner compared with the uninfected HF cells whose expression of GAPDH mRNA did not change much. CMV particles were found
with electron microscope in the cells. Microfilaments were ruptured and shortened after the infection of CMV. Conclusion:
CMV can not only infect human embryo fibroblast cells line HF cells and replicate in the cells, but can also affect the expression
of β-actin mRNA and the microfilaments.
Project (No. 001103058) supported by the Key Program of Science and Technology Bureau of Zhejiang Province, China 相似文献
11.
目的 :研究新城疫病毒在体外抗胃癌细胞活性及其与细胞凋亡的关系。方法 :应用倒置显微镜观察细胞形态、MTT法测NDV在体外对BGC - 82 3的抑制和杀伤作用 ,同时用流式细胞术检测胃癌细胞凋亡情况及细胞分裂周期各时象的变化。结果 :NDV在体外可使BGC - 82 3胃癌细胞形成明显的细胞病变效应、细胞生长抑制及细胞凋亡 ,且细胞凋亡率与感染时间呈正相关。G2、S期细胞减少 ,增殖指数 (PI)降低 ,与阴性对照组比较有显著性差异 (P<0 .0 1)。结论 :NDV具有显著的抗BGC - 82 3胃癌细胞活性。 相似文献
12.
目的检测复发性自然流产（RSA）患者血清RLX-2,TIMP1,TIMP2的表达水平,探讨RLX-2,TIMPs在RSA发生过程中的作用机制,及其对RSA的临床诊断价值。方法采用ELISA法对18例RSA患者与18例正常妊娠者不同妊娠期血清RLX-2,TIMP1,TIMP2的表达水平进行检测,采用受试者工作特性曲线对血清RLX-2,TIMP2及RLX-2/TIMP-2比值在RSA临床诊断的价值进行评价。结果血清RLX-2的水平在妊娠呈上升趋势,10～12周达到高峰;实验组血清TIMP-2水平在各妊娠期均高于正常组（P〈0.05）,各妊娠期RLX-2/TIMP-2比值的曲线下面积（AUC）均较RLX-2,TIMP-2高,其最佳拐点为≤0.1（P〈0.000 1）。结论RSA的发生可能与妊娠期RLX-2的分泌不足以及过高的血清TIMP-2浓度有关;妊娠10～12周时RLX-2/TIMP-2比值≤0.1对RSA的临床诊断价值最大。 相似文献
13.
目的探讨银杏叶总黄酮对体外培养的人肝癌细胞HepG2增殖与凋亡的影响。方法将银杏叶总黄酮作用于体外培养的人肝癌细胞HepG2,MTT法检测其对HepG2细胞增殖的影响,缺口末端核苷标记（TUNNEL）法检测其对HepG2细胞凋亡的影响。结果银杏叶总黄酮对体外培养的人肝癌细胞HepG2的增殖效率下降,使凋亡细胞数增加（P〈0．01）,且呈剂量依赖效应。结论银杏叶总黄酮对体外培养的人HepG2细胞增殖有抑制作用,并能诱导细胞凋亡。 相似文献
14.
目的探讨银杏叶黄酮单体成分槲皮素体外对人肝癌细胞HepG2增殖与凋亡的影响。方法将银杏叶黄酮单体成分槲皮素作用于体外培养的人肝癌细胞HepG2,MTT法检测其对HepG2细胞增殖的影响,缺口末端核苷标记（TUNNEL）法检测其对HepG2细胞凋亡的影响。结果银杏叶黄酮单体成分槲皮素使体外培养的人肝癌细胞HepG2的增殖效率下降,使凋亡细胞数增加（P〈0.01）,两者均呈剂量依赖效应。结论银杏叶黄酮单体成分槲皮素具有与银杏叶总黄酮相似的作用,对体外培养的人HepG2细胞增殖有抑制作用,并能诱导细胞凋亡。 相似文献
15.
目的：探讨增强型绿色荧光蛋白（enhanced green fluorescent protein, EGFP）在中华仓鼠卵巢细胞（CHO-DHFR）细胞中的作用。方法：将增强型绿色荧光蛋白基因的真核表达载体pcDNA3．1（＋）-EGFP，转染至培养的中华仓鼠卵巢细胞（Chinese Hamster Ovary, CHO-DHFR^- ）中。结果：成功表达并产生绿色荧光。结论：证明EGFP是一种良好的报告基因和筛选标志．为进一步研究应用最广泛的哺乳动物细胞表达系统一CHO细胞表达系统奠定了基础。 相似文献
16.
A novel arterial pouch model of saccular aneurysm by concomitant elastase and collagenase digestion 总被引：2，自引：0，他引：2
Background: An ideal aneurysm model of cerebral aneurysm is of great importance for studying the pathogenesis of the lesion and testing new techniques for diagnosis and treatment. Several models have been created in rabbits and are now widely used in experimental studies; however, every model has certain intrinsic limitations. Here we report the development of a novel saccular aneurysm model in rabbits using an arterial pouch that is subject to in vitro pre-digestion with combined elastase and collagenase. Methods: A segment of right common carotid artery (CCA) was dissected out and treated with elastase (60 U/ml, 20 min) followed by type I collagenase (1 mg/ml, 15 min) in vitro. The graft was anastomosed to an arterial arch built with the left CCA and the remaining right CCA, while the other end of the graft was ligated. The dimension and tissue structure of the pouch were analysed immediately, 2 or 8 weeks after operation. Findings: Ten terminal aneurysms were produced. The gross mor-phology of the aneurysm resembles the human cerebral terminal aneurysms. We have observed the following pathological changes: (1) growth of the aneurysm (mean diameter increased from (2.0±0.1) to (3.2±0.3) mm at 2 weeks, P<0.001, n=7~10); (2) thinning of the aneurysmal wall (the mean wall thickness decreased to 44% at 2 weeks), which was accompanied by significant losses of elastic fibres, collagen and the cellular component; and (3) spontaneous rupture (3 out of 9, one aneurysm ruptured 24 h after operation with the other two at 2 and 4 weeks respectively). Conclusion: This rabbit arterial pouch model mimics human cerebral aneurysms in relation to morphology and histology. In particular, this model exhibited an increased tendency of spontaneous rupture. 相似文献
17.
在人体内,CCR5与许多免疫疾病有关,CCR5有望成为众多药物的作用靶点。将ccr5基因与真核表达载体pBBS242构建成重组质粒pBBS242-ccr5,转染CHO细胞,并经潮霉素B筛选。流式细胞仪检测结果表明CCR5在CHO细胞得到了稳定表达。 相似文献
18.
This study aimed to explore the molecular mechanism in tumor invasion and metastasis. The expression of matrix metalloproteinase-2,
−9 (MMP-2, MMP-9), tissue inhibitor-1 of matrix metalloproteinase (TIMP-1), cell adhesion molecule 44 variant 6 (CD44v6),
HER2/neu and p53 was investigated in 154 patients with head and neck squamous cell carcinoma (SCC) by ABC and ImmunoMax immunohistochemical
method. Their clinical relevance and correlation were analysed. The expression of MMP-2, MMP-9, TIMP-1, CD44v6, HER2/neu and
p53 was found in cancer cells in 87.01%, 85.71%, 68.18%, 98.05%, 55.19% and 50.65% cases respectively. Linear regression and
correlation analysis revealed that there was close positive relationship (P<0.05) between the expression of MMP-2 and MMP-9, TIMP-1 and CD44v6, HER2/neu and MMP-9, MMP-2 and p53. Up-regulation of MMP-2
was accompanied by advanced T stage (P<0.01). There was also a trend of MMP-2 expression being related with tumor metastasis. Increased expression of HER2/neu was
found in patients with tumor recurrence (P<0.05). The expression of TIMP-1 was higher in laryngeal cancer than that in pharyngeal cancer, and higher in keratinizing
and non-keratinizing SCC than that in basaloid SCC (P<0.05). These findings suggested that MMP-2 and MMP-9, HER2/neu and MMP-9, MMP-2 and p53 had a coordinate function in aggression
of tumor; that MMP-2 had a more important function than MMP-9 in tumor invasion and metastasis; and that HER2/neu might serve
as a biomarker for poor prognosis in HNSCC.
Project supported by Lübeck Medical University, Germany 相似文献
19.
20.
目的:研究新城疫病毒(NDV)感染对小鼠肉瘤S180细胞p53蛋白的表达及细胞周期的影响。方法:通过NDV体外感染小鼠肉瘤S180细胞,经倒置显微镜观察细胞形态变化,噻唑蓝(MTT)比色法检测小鼠肉瘤S180细胞的增殖状况;经流式细胞仪分析检测NDV体内感染后荷瘤鼠腹水中S180细胞分裂周期各时相的变化、细胞凋亡情况及细胞表面p53蛋白的表达情况。结果:NDV体内、外感染对小鼠肉瘤S180细胞的杀伤作用明显,NDV体内感染后荷瘤鼠腹水中S180细胞高表达p53蛋白,细胞凋亡率增加,G2/S期细胞减少,增殖指数(PI)降低,与阴性对照组比较有显著性差异(P<0.05)。结论:NDV感染可增强小鼠肉瘤S180细胞p53蛋白的表达,影响其细胞周期,诱导其凋亡且有较强的杀瘤作用。 相似文献