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Cell Counting Kit-8 |
产品编号: C0038
产品包装: 500次
产品价格: 612.00元 |
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产品简介
产品简介:
Cell Counting Kit-8,简称CCK-8试剂盒或CCK8试剂盒,是一种基于WST-8而广泛应用于细胞增殖和细胞毒性的快速、高灵敏度检测的试剂盒。
WST-8是一种类似于MTT的化合物,在电子耦合试剂存在的情况下,可以被线粒体内的一些脱氢酶还
原生成橙黄色的formazan(参考图1)。细胞增殖越多越快,则颜色越深;细胞毒性越大,则颜色越浅。对于同样的细胞,颜色的深浅和细胞数目呈线性关系。
图1. WST-8检测原理图 (EC=electron coupling reagent,即电子耦合试剂)
WST-8是MTT的一种升级替代产品,和MTT或其它MTT类似产品如XTT、MTS等相比有明显的优点。首先,MTT被线粒体内的一些脱氢酶还原生成的formazan不是水溶性的,需要有特定的溶解液来溶解;而WST-8和XTT、MTS产生的formazan都是水溶性的,可以省去后续的溶解步骤。其次,WST-8产生的formazan比XTT和MTS产生的formazan更易溶解。再次,WST-8比XTT和MTS更加稳定,使实验结果更加稳定。另外,WST-8和MTT、XTT等相比线性范围更宽,灵敏度更高。
WST-8和WST-1相比,检测灵敏度更高,更易溶解,并且更加稳定。
本试剂盒可以用于细胞因子等诱导的细胞增殖检测,也可以用于抗癌药物等对细胞有毒试剂诱导的细胞毒性检测,或一些药物诱导的细胞生长抑制检测。
本试剂盒检测非常便捷。试剂盒仅一管已经配制好的含有WST-8的CCK-8溶液,无须再进行任何配制等操作。无须使用同位素,所有的检测步骤仅在同一块96孔板内完成。不必洗涤细胞,不必收集细胞,也不必采用额外的步骤去溶解formazan。可以用于大批量样品的检测。
酚红和血清对本试剂盒的测定无明显影响。
WST-8对细胞无明显毒性。加入CCK-8溶液显色后,可以在不同时间反复用酶标仪读板,使检测时间更加灵活,便于找到最佳测定时间。
关于不同的细胞增殖和细胞毒性检测试剂盒的比较和选择,请参考http://www.beyotime.com/mtt-comp.htm。
本试剂盒可以测定500个样品。
包装清单:
产品编号 |
产品名称 |
包装 |
C0038 |
CCK-8溶液 |
5ml |
— |
说明书 |
1份 |
保存条件:
4℃避光保存一年有效,-20℃避光保存两年有效。
注意事项:
由于使用96孔板进行检测,如果细胞培养时间较长,一定要注意蒸发问题。一方面,由于96孔板周围一圈最容易蒸发,可以采取弃用周围一圈的办法,改加相同量的PBS、水或培养液;另一方面,可以把96孔板置于靠近培养箱内水源的地方,以缓解蒸发。
本试剂盒的检测依赖于脱氢酶催化的反应,所以还原剂(例如一些抗氧化剂)会干扰检测,如果待检测体系中存在较多的还原剂,需设法去除。
用酶标仪检测前需确保每个孔内没有气泡,否则会干扰测定。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。
使用说明
使用说明:
1. 通常细胞增殖实验每孔加入100微升2000个细胞,细胞毒性实验每孔加入100微升5000个细胞(具体每孔所
用的细胞的数目,需根据细胞的大小、细胞增殖速度的快慢等因素决定)。按照实验需要,进行培养并给
予0-10微升特定的药物刺激。
2. 每孔加入10微升CCK-8溶液。如果起始的培养体积为200微升,则需加入20微升CCK-8溶液,其它情况以此
类推。可以用加了相应量细胞培养液和CCK-8溶液但没有加入细胞的孔作为空白对照。如果担心所使用的
药物会干扰检测,需设置加了相应量细胞培养液、药物和CCK-8溶液但没有加入细胞的孔作为空白对照。
3. 在细胞培养箱内继续孵育0.5-4小时,对于大多数情况孵育1小时就可以了。时间的长短根据细胞的类型
和细胞的密度等实验情况而定,初次实验时可以在0.5、1、2和4小时后分别用酶标仪检测,然后选取吸
光度范围比较适宜的一个时间点用于后续实验。
4. 在450nm测定吸光度。如无450nm滤光片,可以使用420-480nm的滤光片。可以使用大于600nm的波长,例
如650nm,作为参考波长进行双波长测定。
常见问题:
1. CCK-8溶液对细胞的毒性大小如何?
CCK-8溶液对细胞的毒性非常低,通常观察不到细胞毒性。细胞在CCK-8法检测后仍然可以正常生长,并
可以用于其它的细胞实验。但为了避免CCK-8溶液可能带来的对于后续检测的影响,除非该细胞极难获
得,否则不推荐把CCK-8溶液孵育过的细胞用于其它实验。
2. 如果吸光度值太低,可以采取什么办法?
a. 适当增加细胞数量。
b. 延长加入CCK-8试剂后的孵育时间。
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Overexpression of Dickkopf-3 induces apoptosis through mitochondrial pathway in human
colon cancer.
World J Gastroenterol. 2012 Apr 14;18(14):1590-601. doi: 10.3748/wjg.v18.i14.1590.
72.Qing H, Gong W, Che Y, Wang X, Peng L, Liang Y, Wang W, Deng Q, Zhang H, Jiang B.
PAK1-dependent MAPK pathway activation is required for colorectal cancer cell
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Tumour Biol. 2012 Aug;33(4):985-94. doi: 10.1007/s13277-012-0327-1. Epub 2012 Jan 18.
73.Su Y, Bi J, Wang Y, Tan Y, Yang J, Liu H, Gu W.
Chemical constituents from Chirita longgangensis var. hongyao with inhibitory activity
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74.Wang M, Liu X, Cao C, Wang L.
Highly luminescent CuInS2–ZnS nanocrystals: achieving phase transfer and nuclear homing
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75.Shen Y, Zhao L, Qi L, Qiao J, Mao L, Chen Y.
Reactive polymer as a versatile toolbox for construction of multifunctional
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76.Li XX, He GR, Mu X, Xu B, Tian S, Yu X, Meng FR, Xuan ZH, Du GH.
Protective effects of baicalein against rotenone-induced neurotoxicity in PC12 cells
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Eur J Pharmacol. 2012 Jan 15;674(2-3):227-33. doi: 10.1016/j.ejphar.2011.09.181.
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77.Zhong ZN, Zhu SF, Yuan AD, Lu GH, He ZY, Fa ZQ, Li WH.
Potential of placenta-derived mesenchymal stem cells as seed cells for bone tissue
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78.Feng YJ, Zhang BY, Yao RY, Lu Y.
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Hepatobiliary Pancreat Dis Int. 2012 Aug 15;11(4):418-23.
79.Lin SZ, Wei WT, Chen H, Chen KJ, Tong HF, Wang ZH, Ni ZL, Liu HB, Guo HC, Liu DL.
Antitumor activity of emodin against pancreatic cancer depends on its dual role:
promotion of apoptosis andsuppression of angiogenesis.
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80.Yue X, Zhang Z, Liang X, Gao L, Zhang X, Zhao D, Liu X, Ma H, Guo M, Spear BT, Gong Y,
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81.Shi S, Jia J, Guo X, Zhao Y, Liu B, Chen D, Guo Y, Zhang X.
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82.Yang ZR, Liu M, Peng XL, Lei XF, Zhang JX, Dong WG.
Noscapine induces mitochondria-mediated apoptosis in human colon cancer cells in vivo
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83.Tu Z, Li H, Ma Y, Tang B, Tian J, Akers W, Achilefu S, Gu Y.
The enhanced antiproliferative response to combined treatment of trichostatin A with
raloxifene in MCF-7 breast cancer cells and its relevance to estrogen receptor β
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Mol Cell Biochem.2012 Jul;366(1-2):111-22.doi: 10.1007/s11010-012-1288-9.
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84.Wang M, Liu X, Cao C, Shi C.
Synthesis of band-gap tunable Cu–In–S ternary nanocrystals in aqueous solution.
RSC Advances.Issue 7, 2012 ,2, 2666-2670.
85.Wu D, Guo Z, Min W, Zhou B, Li M, Li W, Luo D.
Upregulation of TCTP expression in human skin squamous cell carcinoma increases tumor
cell viability throughanti-apoptotic action of the protein.
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86.He H, Yang T, Jia S, Zhang R, Tu P, Gao J, Yuan Y, Han W, Yu Y.
Expression and purification of bioactive high-purity human S100A6 in Escherichia coli.
Protein Expr Purif. 2012 May;83(1):98-103. doi: 10.1016/j.pep.2012.03.002.
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87.Shao X, Hu Z, Hu C, Bu Q, Yan G, Deng P, Lv L, Wu D, Deng Y, Zhao J, Zhu R, Li Y, Li H,
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Taurine protects methamphetamine-induced developmental angiogenesis defect through
antioxidant mechanism.
Toxicol Appl Pharmacol. 2012 May 1;260(3):260-70. doi: 10.1016/j.taap.2012.03.003.
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88.Zhang L, Yang M, Gan L, He T, Xiao X, Stewart MD, Liu X, Yang L, Zhang T, Zhao Y, Fu J.
DLX4 upregulates TWIST and enhances tumor migration, invasion and metastasis.
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89.Sun Q, Kawamura T, Masutani K, Peng X, Sun Q, Stolz DB, Pribis JP, Billiar TR, Sun X,
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Oral intake of hydrogen-rich water inhibits intimal hyperplasia in arterialized vein
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90.Li X, Zheng SL, Li X, Li JL, Qiang O, Liu R, He L.
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91.Ning L, Li X, Ding X, Yin Y, Li G.
An iRGD Based Strategy to Study Electrochemically the Species Inside a Cell.
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92.Zhou X, Meng Q, Xu X, Zhi T, Shi Q, Wang Y, Yu R.
Bex2 regulates cell proliferation and apoptosis in malignant glioma cells via the c-Jun
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94.Wang W, Tao R, Tong Z, Ding Y, Kuang R, Zhai S, Liu J, Ni L.
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Peptides.2012 Feb;33(2):+212-9.doi:10.1016/j.peptides.2012.01.006.Epub 2012 Jan 20.
95.Du X, Zhao B, Li J, Cao X, Diao M, Feng H, Chen X, Chen Z, Zeng X.
Astragalus polysaccharides enhance immune responses of HBV DNA vaccination via promoting
the dendritic cell maturation and suppressing Treg frequency in mice.
Int Immunopharmacol. 2012 Dec;14(4):463-70. doi: 10.1016/j.intimp.2012.09.006.
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96.Zhou BR, Yin HB, Xu Y, Wu D, Zhang ZH, Yin ZQ, Permatasari F, Luo D.
Baicalin protects human skin fibroblasts from ultraviolet A radiation-induced oxidative
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Free Radic Res. 2012 Dec;46(12):1458-71. doi: 10.3109/10715762.2012.726355.
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97.Wu Q, Wang C, Guo L, Ge Q, Lu Z.
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98.Hu Y, Qi Y, Liu H, Fan G, Chai Y.
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99.Shen XL, Zhang Y, Xu W, Liang R, Zheng J, Luo Y, Wang Y, Huang K.
An iTRAQ-based mitoproteomics approach for profiling the nephrotoxicity mechanisms of
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101.Guo Z, Zhou B, Liu W, Xu Y, Wu D, Yin Z, Permatasari F, Luo D.
MiR-23a regulates DNA damage repair and apoptosis in UVB-irradiated HaCaT cells.
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102.Ma GF, Chen SY, Sun ZR, Miao Q, Liu YM, Zeng XQ, Luo TC, Ma LL, Lian JJ, Song DL.
FoxP3 inhibits proliferation and induces apoptosis of gastric cancer cells by activating
the apoptotic signalingpathway.
Biochem Biophys Res Commun.2013 Jan 11;430(2):804-9.doi:10.1016/j.bbrc.2012.11.065.
103.Zhao L, Wang H, Huo K, Zhang X, Wang W, Zhang Y, Wu Z, Chu PK.
The osteogenic activity of strontium loaded titania nanotube arrays on titanium
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104.Pang H, Wu XH, Fu SL, Luo F, Zhang ZH, Hou TY, Li ZQ, Chang ZQ, Yu B, Xu JZ.
Co-culture with endothelial progenitor cells promotes survival, migration, and
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105.Gao C, Cai Y, Kong X, Han G, Yao J.
Development and characterization of injectable chitosan-based hydrogels containing
dexamethasone/rhBMP-2 loaded hydroxyapatite nanoparticles.
Materials Letters.Volume 93, 15 February 2013, Pages 312–315.
106.Chang Y, Huang X, Liu Z, Han G, Huang L, Xiong YC, Wang Z.
Dexmedetomidine inhibits the secretion of high mobility group box 1 from
lipopolysaccharide-activatedmacrophages in vitro.
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107.Wang Y, Wang K, Li H, Chen L, Xu F, Wu T.
Effects of different sustained hydrostatic pressures on connexin 43 in human bladder
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108.Li Y, Wang Y, Wang P, Zhang B, Yan W, Sun J, Pan J.
In vitro cytocompatibility evaluation of MGF-Ct24E chemically grafted and physically
blended with maleicanhydride modified poly(D, L-lactic acid).
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109.Alibrahim A, Zhao LY, Bae CY, Barszczyk A, Sun CL, Wang GL, Sun HS.
Neuroprotective effects of volume-regulated anion channel blocker DCPIB on neonatal
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Acta Pharmacol Sin. 2013 Jan;34(1):113-8. doi: 10.1038/aps.2012.148. Epub 2012 Dec 3.
110.Shu G, Xie B, Ren F, Liu DC, Zhou J, Li Q, Chen J, Yuan L, Zhou J.
Restoration of klotho expression induces apoptosis and autophagy in hepatocellular
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111.Kuang P, Zhao W, Su W, Zhang Z, Zhang L, Liu J, Ren G, Yin Z, Wang X.
18β-glycyrrhetinic acid inhibits hepatocellular carcinoma development by reversing
hepatic stellate cell-mediatedimmunosuppression in mice.
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112.Liu Z, Chang Y, Zhang J, Huang X, Jiang J, Li S, Wang Z.
Magnesium deficiency promotes secretion of high-mobility group box 1 protein from
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113.Zhang M, Qiu Z, Li Y, Yang Y, Zhang Q, Xiang Q, Su Z, Huang Y.
Construction and characterization of a recombinant human beta defensin 2 fusion protein
targeting the epidermalgrowth factor receptor: in vitro study.
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114.Ji X, Lu W, Zhou H, Han D, Yang L, Wu H, Li J, Liu H, Zhang J, Cao P, Zhang S.
Covalently dimerized Camelidae antihuman TNFa single-domain antibodies expressed in
yeast
Pichia pastorisshow superior neutralizing activity.
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115.Shan D, Shi Y, Duan S, Wei Y, Cai Q, Yang X.
Electrospun magnetic poly(l-lactide) (PLLA) nanofibers by incorporating PLLA-stabilized
Fe3O4 nanoparticles.
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116.Xie H, Li J, Li L, Dong Y, Chen GQ, Chen KC.
Enhanced proliferation and differentiation of neural stem cells grown on PHA films
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FIM-A, a Phosphorus-Containing Sirolimus, Inhibits the Angiogenesis and Proliferation of
Osteosarcomas.
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118.Wang H, Jiang W, Yuan L, Wang L, Chen H.
Reductase-like activity of silicon nanowire arrays.
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119.Zhan XX, Liu Y, Yang JF, Wang GY, Mu L, Zhang TS, Xie XL, Wang JH, Liu YM, Kong QF,
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BRG1 is a prognostic marker and potential therapeutic target in human breast cancer.
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121.Feng H, Du X, Tang J, Cao X, Han X, Chen Z, Chen Y, Zeng X.
Enhancement of the immune responses to foot-and-mouth disease vaccination in mice by
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122.Yang Q, Sui G, Shi Y, Duan S, Bao J, Cai Q, Yang X.
Osteocompatibility characterization of polyacrylonitrile carbon nanofibers containing
bioactive glass nanoparticles.
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123.Hu J, Zhang L, Wang N, Ding R, Cui S, Zhu F, Xie Y, Sun X, Wu D, Hong Q, Li Q, Shi S,
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Mesenchymal stem cells attenuate ischemic acute kidney injury by inducing regulatory T
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124.Wang S, Pang T, Gao M, Kang H, Ding W, Sun X, Zhao Y, Zhu W, Tang X, Yao Y, Hu X.
HPV E6 induces eIF4E transcription to promote the proliferation and migration of
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125.Yang L, Zhou L, Zong X, Cao X, Ji X, Gu W, Zhang S.
Characterization of the molecular structure, expression and bioactivity of the TNFSF13B
(BAFF) gene of the SouthAfrican clawed frog, Xenopus laevis.
Int Immunopharmacol.2013 Mar;15(3):478-87.doi:10.1016/j.intimp.2013.02.006.
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126.Wu K, Song W, Zhao L, Liu M, Yan J, Andersen MØ, Kjems J, Gao S, Zhang Y.
MicroRNA functionalized microporous titanium oxide surface by lyophilization with
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127.Yao W, Cai X, Liu C, Qin Y, Cheng H, Ji S, Xu W, Wu C, Chen T, Xu J, Long J, Fang Z,
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Profilin 1 potentiates apoptosis induced by staurosporine in cancer cells.
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Cullin1 is a novel marker of poor prognosis and a potential therapeutic target in human
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129.Zou X, Yu C, Zhang H.
In Vitro Analyses of Effect of Light on Toxicity of Titanium Dioxide Nanoparticles to
Tetrahymena Pyriformis Using Fourier Transform Infrared Spectra.
An International Journal for Rapid Communication. Volume 46, Issue 8, 2013.
130.Xu D, Yang W, Hu Y, Luo Z, Li J, Hou Y, Liu Y, Cai K.
Surface functionalization of titanium substrates with cecropin B to improve their
cytocompatibility and reduceinflammation responses.
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131.Shen Z, Jiang X, Zeng C, Zheng S, Luo B, Zeng Y, Ding R, Jiang H, He Q, Guo J, Jie W.
High expression of ubiquitin-conjugating enzyme 2C (UBE2C) correlates with
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132.Jia D, Duan F, Peng P, Sun L, Liu X, Wang L, Wu W, Ruan Y, Gu J.
Up-regulation of RACK1 by TGF-β1 promotes hepatic fibrosis in mice.
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133.Zhu C, Qin H, Cheng T, Tan HL, Guo YY, Shi SF, Chen DS, Zhang XL.
Staphylococcus aureus supernatant induces the release of mouse β-defensin-14 from
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134.Zhang X, Cai Z, Wang G, Wang H, Liu Z, Guo X, Yang C, Wang X, Wang H, Shu C, Xiao L.
F-actin may play an important role in IL-1β-stimulated hippocampal neurons.
Behav Brain Res.2013 Apr15;243:165-70.doi:10.1016/j.bbr.2013.01.001.Epub 2013 Jan 11.
135.Wang Q, Shen L, Huang W, Song Y, Xiao L, Xu W, Liu Y.
Vasculogenesis of decidua side population cells of first-trimester pregnancy.
Stem Cell Res Ther. 2013 May 7;4(3):50. [Epub ahead of print].
136.Zhen Y, Ye Y, Yu X, Mai C, Zhou Y, Chen Y, Yang H, Lyu X, Song Y, Wu Q, Fu Q, Zhao M,
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Reduced CTGF Expression Promotes Cell Growth, Migration, and Invasion in Nasopharyngeal
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PLoS One. 2013 Jun 3;8(6):e64976. doi: 10.1371/journal.pone.0064976. Print 2013.
137.Shi SF, Zhang XL, Zhu C, Chen DS, Guo YY.
Ultrasonically enhanced rifampin activity against internalized Staphylococcus aureus.
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138.Duan S, Yang X, Mao J, Qi B, Cai Q, Shen H, Yang F, Deng X, Wang S.
Osteocompatibility evaluation of poly (glycine ethyl ester-co-alanine ethyl
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139.Li G, Liu Y, Su Z, Ren S, Zhu G, Tian Y, Qiu Y.
MicroRNA-324-3p regulates nasopharyngeal carcinoma radioresistance by directly
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140.Jiao Q, Wang H, Hu Z, Zhuang Y, Yang W, Li M, Yu X, Liang J, Guo Y, Zhang H, Chen X,
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Lidocaine inhibits staphylococcal enterotoxin-stimulated activation of peripheral blood
mononuclear cells frompatients with atopic dermatitis.
Arch Dermatol Res. 2013 Apr 16. [Epub ahead of print].
141.Xu Y, Wu J, Wang H, Li H, Di N, Song L, Li S, Li D, Xiang Y, Liu W, Mo X, Zhou Q.
Fabrication of Electrospun Poly(L-Lactide-co-ε-Caprolactone)/Collagen Nanoyarn Network
as a Novel, Three-Dimensional, Macroporous, Aligned Scaffold for Tendon Tissue
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142.Lv L, Liu H, Chen X, Yang Z.
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143.Xia J, Wang F, Wang L, Fan Q.
Elevated serine protease HtrA1 inhibits cell proliferation, reduces invasion, and
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144.Jiang ZQ, Yan XJ, Bi L, Chen JP, Zhou Q, Chen WP.
Mechanism for hepato-protective action of Liangxue Huayu Recipe (LHR): Blockade of
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145.Wu S, Ju GQ, Du T, Zhu YJ, Liu GH.
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146.Yang C, Wu D, Jia J, Liu D, Li Z, Zhang C, Li M, Xia Y.
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147.Zhou X, Qian J, Hua L, Shi Q, Liu Z, Xu Y, Sang B, Mo J, Yu R.
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148.Jin Y, Peng D, Shen Y, Xu M, Liang Y, Xiao B, Lu J.
MicroRNA-376c Inhibits Cell Proliferation and Invasion in Osteosarcoma by Targeting to
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DNA Cell Biol. 2013 Jun;32(6):302-9. doi: 10.1089/dna.2013.1977. Epub 2013 Apr 30.
149.Hu X , Liu R, Zhu N.
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Immunobiology.Available online 30 May 2013.
150.Xia L, Huang Q, Shi J, Nie D, Gong M, Gong P, Zhao L, Zuo H, Huang H, Ju S, Chen J,
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PAX3 is overexpressed in human glioblastomas and critically regulates the tumorigenicity
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151.Han Y, Guo XH, Zheng QF, Zhu YL, Fan YY, Zhang XY.
Down-regulation of platelet-derived growth factor-D expression blockades NF-κB pathway
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152.Wang L, Liang B, Li L, Liu W.
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153.Gao F, Ding B, Zhou L, Gao X, Guo H, Xu H.
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154.Chen Q, Xue Y, Sun J.
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