舒尼替尼对胃癌细胞BGC-823增殖的影响及机制

探索舒尼替尼对胃癌细胞BGC-823增殖的影响及机制。不同浓度舒尼替尼处理BGC-823细胞24、48、72、96 h后,MTT法检测细胞增殖,得出药物的半数抑制浓度为0.592 8μmol/L。筛选出比较接近半数致死量的3个药物浓度0.25、0.50、1.00μmol/L作用细胞48 h后,细胞核染色检测细胞凋亡;免疫荧光染色法检测Notch-1的表达及细胞凋亡水平;蛋白质免疫印迹检测Notch-1、caspase-3、caspase-9蛋白表达水平。MTT检测结果显示,舒尼替尼在浓度0.156 25～10μmol/L时可显著抑制BGC-823细胞增殖,且具有时间和剂量依赖性。细胞核染色结果显示,不同浓度的舒尼替尼处理BGC-823细胞48 h后随浓度的增加,细胞凋亡数量显著增多。蛋白质免疫印迹结果显示,不同浓度舒尼替尼作用48 h后,Notch-1蛋白的表达水平随浓度的增加而降低,caspase-3、caspase-9表达水平随浓度的增加而升高。舒尼替尼通过抑制Notch-1的信号通路活性,进而抑制BGC-823细胞的增殖并诱导细胞凋亡。     

舒尼替尼对转移性肾癌患者髓系来源抑制细胞的影响

舒尼替尼是一种口服的多靶点酪氨酸激酶抑制剂,具有抗血管生成和抑制肿瘤增殖的作用.与IFN-α相比,舒尼替尼明显提高了转移性肾癌(mRCC)患者的无进展生存期,并成为治疗mRGC的一线治疗方案.近期有研究报道,舒尼替尼还可以通过抑制髓系来源抑制细胞(MDSC)调节肿瘤免疫.MDSC是一群来源于髓系细胞谱系的异质细胞,其通...     

酪氨酸激酶抑制剂甲磺酸伊马替尼对PTEN介导的K562细胞侵袭的影响

目的:研究酪氨酸激酶抑制剂甲磺酸伊马替尼对K562细胞PTEN信号转导的调控,以及对细胞侵袭功能的影响.方法:不同浓度甲磺酸伊马替尼作用K562细胞不同时间后,通过荧光定量PCR检测BCR/ABL、PTEN、FAK水平变化及相互关系,免疫细胞化学染色检测FAK蛋白水平,Transwell小室检测K562细胞侵袭功能.结果:2μg/mL甲磺酸伊马替尼作用K562细胞在36 h内,随着BCR/ABL融合基因表达减低,PTEN mRNA表达上调,FAK mRNA及蛋白表达下调,K562细胞侵袭功能明显减弱.作用48 h后,随着BCR/ABL融合基因的抑制减弱,PrEN表达进而减低,而FAK表达升高.BCR/ABL mRNA与PTEN mRNA呈负相关趋势,与FAK mRNA呈正相关趋势.结论:甲磺酸伊马替尼通过抑制BCR/ABL融合基因调控PTEN/FAK信号转导通路,参与抑制白血病K562细胞侵袭作用.     

紫草素逆转由肝细胞生长因子诱导的肺癌吉非替尼耐药

目的:研究肝细胞生长因子(hepatocyte growth factor,HGF)诱导人肺癌HCC827细胞对吉非替尼的耐药以及紫草素(shikonin)逆转此耐药作用的可能机制。方法:体外培养HCC827细胞,分别采用不同浓度的紫草素和吉非替尼单独/联合作用,MTT法检测细胞活力,并计算吉非替尼的半数抑制浓度(half maximal inhibitory concentration,IC_(50)); Transwell小室实验检测紫草素对HGF诱导的吉非替尼耐药HCC827细胞侵袭能力的影响; Western blot检测HGF诱导的吉非替尼耐药HCC827细胞中上皮间质转化(EMT)及相关信号通路蛋白水平的变化。结果:随着给药剂量的增加,紫草素对HCC827细胞的生长抑制率显著上升(P 0. 05),并呈一定的剂量依赖关系,其IC_(50)为3. 06μmol/L。肺癌吉非替尼敏感细胞HCC827对吉非替尼的IC_(50)为0. 51μmol/L。利用不同浓度的HGF诱导吉非替尼耐药,发现20μg/L HGF诱导耐药最为明显,在外源性HGF存在的情况下,HCC827细胞对吉非替尼的IC_(50)为12. 71μmol/L。紫草素能逆转由HGF诱导的吉非替尼耐药(P 0. 05)。Transwell小室实验结果显示,与对照组相比,HGF组能显著提高肺癌HCC827细胞的侵袭能力;同时,紫草素联合吉非替尼作用组则明显抑制由HGF诱导的侵袭(P 0. 01)。Western blot实验结果显示,HGF可诱导HCC827细胞发生EMT,使细胞中E-cadherin蛋白表达下调,vimentin蛋白表达上调,紫草素则能逆转EMT的发生(P 0. 01)。此外,HGF组可激活细胞中AKT蛋白的磷酸化;紫草素联合吉非替尼作用组则能明显抑制由HGF激活的AKT蛋白磷酸化水平(P 0. 01)。结论:紫草素能逆转由HGF诱导肺癌HCC827细胞的吉非替尼耐药,其分子机制可能与逆转EMT和抑制AKT蛋白通路的磷酸化水平有关。     

苹果酸舒尼替尼抑制膀胱癌T24细胞系的体外增殖

 摘要：目的：探讨苹果酸舒尼替尼对人膀胱癌T24细胞系体外增殖的影响。方法：取人膀胱癌T24细胞系悬液, 分别加入浓度为0.625、1.25、2.5、5、10 和 20μmol/L的苹果酸舒尼替尼,分别于24、48、72 h进行处理,用细胞毒抑制实验（MTT法）检测药物对其生长的影响。结果：苹果酸舒尼替尼可浓度依赖性抑制人膀胱癌细胞系T24细胞增殖,当药物浓度>5μmol/L时,苹果酸舒尼替尼与顺铂比较,对T24细胞系具有更强的抑制作用（P<0.05）。这种抑制作用随着药物培育时间延长（24、48、72ｈ）而增强,表现为IC50值显著降低, 但其抑制能力与顺铂比较无显著差别（P>0.05）。结论：苹果酸舒尼替尼可呈浓度和时间依赖地抑制膀胱癌T24细胞系增殖。     

PI3K / Akt / mTOR 信号通路介导的N822K 突变对c-KIT 抑制剂诱导的AML 细胞凋亡的影响

目的:研究c-KIT N822K 突变对c-KIT 抑制剂诱导AML 细胞凋亡的影响,并初步探讨相关的分子机制。方法:以c-KIT N822K 突变的Kasumi-1 细胞为实验组,以HL-60、NB4 细胞为非c-KIT N822K 突变的对照组,分别用0、0.04、0.16、0.64 μmol/ L 的c-KIT 抑制剂舒尼替尼处理这三株AML 细胞24 h 后收集细胞,采用Western blot 检测凋亡相关蛋白和PI3K/ Akt/ mTOR 通路蛋白水平,比较各组细胞相关信号通路蛋白的变化。结果:随着舒尼替尼浓度的增加,HL-60 及NB4 细胞中Bax 及CytoC、Caspase-9、Actived-Caspase-3、PARP 蛋白剪切体等促凋亡相关蛋白表达均上调(P<0.05),抗凋亡蛋白Bcl-2表达均下调(P<0.01),在具有N822K 突变的Kasumi-1 细胞中这一变化趋势则明显减弱;Kasumi-1 细胞中c-myc 蛋白及PI3K、Akt、4EBP1、mTOR 等PI3K/ Akt/ mTOR 通路蛋白的磷酸化水平均出现剂量依赖性下调(P<0.05),而HL-60 细胞和NB4 细胞则无此变化。结论:N822K 突变引起的c-KIT 结构性激活可影响c-KIT 抑制剂舒尼替尼对Kasumi-1 细胞的凋亡诱导作用,其机制可能与PI3K/ Akt/ mTOR 通路抑制有关。     

榄香烯通过c-Met信号通路逆转由肝细胞生长因子诱导的吉非替尼耐药作用

目的:研究肝细胞生长因子(hepatocyte growth factor,HGF)诱导肺癌PC-9细胞对吉非替尼耐药的作用,并探讨榄香烯逆转此耐药作用的可能机制。方法:应用不同浓度的吉非替尼和榄香烯单独或联合作用于HGF诱导的PC-9细胞株。MTT法检测细胞活力,并计算吉非替尼的半数抑制浓度(half maximal inhibitory concentration,IC50);Transwell小室实验检测榄香烯对HGF诱导的吉非替尼耐药PC-9细胞侵袭能力的影响;Western blot检测PC-9细胞中c-Met、AKT及其磷酸化蛋白的水平。结果:榄香烯可以显著抑制肺癌PC-9细胞的活力(P0.05),随着药物剂量的增加,榄香烯对PC-9细胞的生长抑制率显著上升,在给药24 h后,其IC50为169.31 mg/L。肺癌PC-9细胞对吉非替尼敏感,随着吉非替尼浓度的增加,其对PC-9细胞生长的抑制作用不断增强,IC50为0.30μmol/L;外源性HGF(50μg/L)对肺癌PC-9细胞的生长有明显的促进作用,并诱导吉非替尼耐药。同时,使用cMet抑制剂SU11274联合吉非替尼作用于HGF诱导的PC-9细胞时,其存活率比吉非替尼单独作用于HGF诱导的PC-9细胞明显降低(P0.05)。Transwell小室结果显示,与对照组相比,HGF能显著提高肺癌PC-9细胞的侵袭能力;与HGF联合吉非替尼组相比,榄香烯、HGF联合吉非替尼作用能明显抑制肺癌PC-9细胞的侵袭能力。Western blot结果显示,与对照组相比,HGF可显著上调p-Met和p-AKT的蛋白水平;与HGF联合吉非替尼组相比,榄香烯、HGF联合吉非替尼作用能明显下调p-Met和p-AKT的蛋白水平(P0.01)。结论:榄香烯可逆转HGF诱导的肺癌PC-9细胞对吉非替尼耐药,其机制可能与抑制HGF活化的c-Met及其下游信号通路有关。     

舒尼替尼抑制小鼠骨髓来源树突状细胞表面PD-L1和PD-L2的表达

目的观察小鼠髓源性树突状细胞(DC)在舒尼替尼刺激下,其表面程序性死亡分子1配体1(PD-L1)和PD-L2的表达变化。方法取小鼠骨髓细胞,对照组加入二甲基亚砜,实验组分别加入(100、200、300)ng/m L舒尼替尼,刺激48 h,用流式细胞术检测DC表面PD-L1和PD-L2的表达水平。结果与对照组相比,实验组中成熟DC(m DC)和总DC(包括m DC和im DC)表面PD-L1的表达水平显著降低;表达PD-L1的未成熟DC(im DC)、m DC和DC百分比均显著降低,表达PD-L2的m DC百分比显著降低;表达PD-L2的DC百分比在100 ng/m L、300 ng/m L舒尼替尼组中显著降低。结论舒尼替尼可显著降低小鼠DC表面PD-L1、PD-L2的表达。     

人肺腺癌厄洛替尼耐药细胞系PC-9/ER的建立及其特性

目的：构建厄洛替尼耐药人肺腺癌细胞模型PC-9/ER,观察单用人表皮生长因子受体(epithelial growth factor receptor,EGFR)酪氨酸激酶抑制剂厄洛替尼或联合胰岛素样生长因子受体1(insulin-like growth factor receptor 1,IGF1R)酪氨酸激酶抑制剂苦鬼臼毒(picropodophyllotoxin,PPP)作用于该细胞后,该细胞对厄洛替尼耐药性的影响,并探讨耐药相关机制。方法：选择人肺腺癌细胞株PC-9,采用逐步递增厄洛替尼浓度的方法体外诱导构建耐药株PC-9/ER。CCK-8法检测耐药指数;细胞计数法绘制PC-9和PC-9/ER的生长曲线,并计算出两细胞系的倍增时间;流式细胞术检测两细胞系的细胞周期;Western Blot法检测p-EGFR及p-IGF1R的表达水平,并进一步检测厄洛替尼及PPP单独或联合作用于PC-9/ER后,各组Akt,ERK,p-Akt及p-ERK的表达水平。结果：PC-9/ER细胞株的耐药指数是72.3。细胞生长曲线显示,PC-9/ER细胞生长较亲代细胞慢,PC-9与PC-9/ER细胞的倍增时间分别为32.9及36.9 h (P=0.003)。与PC-9相比,PC-9/ER细胞的G1期细胞增多(P=0.001),而S期细胞则明显下降(P=0.015)。Western Blot结果表明,PC-9/ER细胞中p-IGF1R表达比亲代细胞明显增多(P=0.016),而p-EGFR无明显变化(P=0.152)。在亲代及耐药细胞系,厄洛替尼联合PPP组均较其他组更能显著的抑制细胞增殖(P<0.05);且Western Blot法表明联合用药组EGFR下游磷酸化的Akt、ERK的表达水平明显减少。结论：成功构建了人肺腺癌厄洛替尼耐药细胞株PC-9/ER,IGF1R通路可能与肺腺癌EGFR-TKI获得性耐药有关。     

舒尼替尼用于治疗晚期肾细胞癌的临床分析

目的 研究晚期肾细胞癌患者应用舒尼替尼进行治疗的临床效果,为今后的治疗提供参考依据。方法 我院2015年10月~2016年10月共收治运用舒尼替尼治疗的晚期肾细胞癌患者36例,用药过程中如患者出现药物耐受力差的现象,则改注射为口服用药。了解分析患者在治疗期间的临床资料,观察舒尼替尼运用在治疗过程中具体用量、疗效及不良反应。结果 舒尼替尼用于治疗晚期肾细胞癌的有效控制率为88.88%。不良反应主要有手足综合征、高血压、困顿乏力、口腔粘膜炎、骨髓抑制以及食欲减退等,在调整药物用量后,患者基本可以忍受。结论 应用舒尼替尼治疗晚期肾细胞癌患者,疾病控制效果较为显著,治疗过程安全可靠,由于用药所引发的不良反应患者表示可以忍受,应用舒尼替尼治疗后,患者有望延长存活期,缓解疾病痛苦。     

An investigation of the fate of cells transplanted orthotopically between morulae/nascent blastocysts in the mouse

A technique has been devised for selectively replacing someinside cells (ICs) or outside cells (OCs) of late morulae/nascentblastocysts with corresponding cells from genetically dissimilar,synchronous embryos. The main purpose was to determine whetherthe inner cell mass (ICM) contributes cells to the overlyingpolar trophectoderm at any stage during the blastocyst phaseof development. Notwithstanding the high incidence and levelof chimaerism in ICM derivatives of postimplantation conceptusesobtained in the IC transplantation experiments, trophoblasttissue was composed entirely of host cells in the majority ofcases. Even where a donor IC contribution to trophoblast wasdetected there were strong grounds for suspecting that it wasdue to tissue contamination rather than genuine chimaerism.Thus, not only did such contributions differ in both level anddistribution from those produced by transplantation of OCs butthey also varied markedly in frequency according to the dayof gestation on which conceptuses were dissected. The possibilitythat ICs regularly colonize the polar trophectoderm but failto persist there was excluded by the results of short-term transplantation experiments using an in-situ genetic marker. These findingsoffer no support for the hypothesis that the ICM serves as apopulation of stem cells for the trophectoderm as well as theprimitive endoderm and ectoderm during normal development. Thefrequency of chimaerism was lower in OC than IC transplantationexperiments. Nevertheless, in a substantial proportion of chimaeras,OCs colonized derivatives of the ICM. This is consistent withevidence from other studies that some outside cells divide differentiallyat 5th cleavage to produce an OC plus an IC rather than twoOCs.     

The effects of progesterone on apoptosis in the human trophoblast-derived HTR-8/SV neo cells

Progesterone (P4) is frequently used in the treatment of threatened abortion, prevention of recurrent miscarriage and threatened preterm labor. However, little is known about the molecular mechanism of P4 in the regulation of extravillous trophoblasts' (EVTs) function. This study was designed to examine the presence of progesterone receptor (PR) in the human trophoblast-derived HTR-8/SV neo cell line, which is a possible model of EVTs, and the effects of P4 on apoptosis in those cells. The HTR-8/SV neo cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 microg/ml streptomycin. When the cell the population reached 50% confluency, the cells were stepped down to serum-free conditions in the presence or absence of graded concentrations of P4 (1, 10 and 100 ng/ml) for 48 h. The cultured cells were used for RT-PCR, terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL) assay, immunocytochemistry and western blot analyses. Immunocytochemistry and western blot analyses revealed that PR was evident in HTR-8/SV neo cells. Compared with untreated cultures, treatment with P4 (10 and 100 ng/ml) resulted in significant decreases in the TUNEL-positive rate, Fas, Fas ligand (Fas-L), caspase-8, caspase-3 and poly (ADP-ribose) polymerase (PARP) expression in HTR-8/SV neo cells, and a significant increase in Bcl-2 expression in those cells. Consistently, Fas mRNA expression in those cells was significantly inhibited by the treatment with 10 ng/ml P4 compared with untreated cultures. This study suggests that PR exists in HTR-8/SV neo cells and that P4 inhibits apoptosis by down-regulating Fas, Fas-L, caspase-8, caspase-3 and PARP expression as well as up-regulating Bcl-2 expression in HTR-8/SV neo cells.     

Stem cell factor/c-kit system in spermatogenesis

One of the major unresolved questions with male infertility is the identification of the molecular origin of a great majority of the spermatogenetic arrests currently diagnosed as idiopathic male infertility. During the past years, several families of regulating factors have been implicated in spermatogenesis defects observed essentially in animal models. Among these factors are signalling molecules, and particularly the stem cell factor (SCF)/c-kit system. The SCF and its receptor c-kit are an appropriate example to illustrate the role of signalling molecules in the physiology and pathology of spermatogenesis. The SCF/c-kit regulates primordial germ cell migration, proliferation and apoptosis during fetal gonadal development. The SCF/c-kit also regulates spermatogonia proliferation in the adult animal. In mutant mice, abnormalities of the SCF/c-kit gene expression, such as gene deletion, point mutation, alternative splicing defect, lead to different types of spermatogenesis alterations (e.g. decrease in primordial germ cell migration, decrease in spermatogonia proliferation). More recently, defects in SCF/c-kit gene expression have also been shown in human testicular dysfunctions. Indeed, a reduction in SCF/c-kit expression has been evidenced in oligozoospermia/azoospermia associated with an increase in the germ cell apoptosis process. In addition, c-kit seems to be a good marker of seminoma testicular tumours. This review reports a large number of data--obtained essentially in animal models--that suggest an important role for the SCF/c-kit system in spermatogenesis and, as a corollary, its potential involvement in spermatogenic defects.     

IDO expression on decidual and peripheral blood dendritic cells and monocytes/macrophages after treatment with CTLA-4 or interferon-gamma increase in normal pregnancy but decrease in spontaneous abortion

Recent data demonstrated that CD4+CD25+ regulatory T (Treg) cells and an enzyme called indoleamine 2,3-dioxygenase (IDO) mediate maternal tolerance to the fetus. Interestingly, Treg cells express the CTLA-4 molecule on their surface, and B7 (CD80/86) ligation by CTLA-4 enhanced IDO activity of dendritic cells (DCs) and monocytes by the induction of interferon gamma (IFN-gamma) production. In this study, we studied the IDO expression on peripheral blood monocytes and decidual monocytes or DCs after treatment with CTLA-4/Fc fusion protein or IFN-gamma using flow cytometry. IDO expressions on both peripheral blood DC and decidual DC and monocytes were up-regulated during normal pregnancy. On the other hand, both IDO expression on DC and monocytes after IFN-gamma treatment or CTLA-4 treatment were decreased in spontaneous abortion cases. The expression of CD86 on peripheral blood and decidual monocytes and DC in spontaneous abortion cases was lower compared with those in normal pregnancy subjects. Also, IFN-gamma production by decidual and peripheral blood mononuclear cells after CTLA-4/Fc treatment in spontaneous abortion cases was significantly lower than those in normal pregnancy subjects. These data suggest that CTLA-4 on Treg cells up-regulates IDO expression on decidual and peripheral blood DC and monocytes by the induction of IFN-gamma production.     

Synergistic effects of snail and quercetin on renal cell carcinoma Caki-2 by altering AKT/mTOR/ERK1/2 signaling pathways

Renal cell carcinoma has become the most common subtype of kidney cancer, and has the highest propensity to manifest as metastatic disease. Because of lack of knowledge in events that correlated with tumor cell migration and invasion, few therapeutic options are available. Therefore, in current study, we explore the anti-tumoral effect of a potential chemopreventive natural product, quercetin, combined with anti-sense oligo gene therapy (inhibiting Snail gene). We found that either one of them had the remarkable effects in suppressing cell proliferation and migration, inducing cell cycle arrest and apoptosis in a ccRCC cell line, Caki-2 cells. The combination of both means provides even strong suppressive effects toward these ccRCC cells. Our study, for the first time, provides the possibility of using a novel treatment for renal cancer, by combining natural product and gene therapy.     

Autoimmune gld mutation uncouples suicide and cytokine/proliferation pathways in activated,mature T cells

Antigen receptor-directed suicide plays an important role in the elimination of potentially autoaggressive immature T cells during thymic differentiation. Here we demonstrate evidence for a second pathway of receptor-directed suicide in mature T cells that is missing in a mutant strain (gld) of mice with an “autoimmune” lymphoproliferative syndrome. The defect is evident within the gld activated T cell and does not require the presence of an antigen-presenting cell for its expression. Receptor-driven suicide is intact in immature T cells of animals with this mutation. These results support the significance of receptor-directed suicide in the mature T cell compartment and suggest that the immune system may use three independent pathways for regulating programmed cell death in shaping and controlling the immune response.     

Possible involvement of crosstalk cell-adhesion mechanism by endometrial CD26/dipeptidyl peptidase IV and embryonal fibronectin in human blastocyst implantation

When human blastocysts hatch through the zona pellucida, gaining the ability to adhere to the endometrium, crosstalk between the embryo and the uterus may represent a successful outcome of their synchronized development and differentiation. CD26/dipeptidyl peptidase IV is known as a marker molecule of the implantation phase endometrium. To study the role of CD26 in implantation, 35 human hatched blastocysts were prepared by enzymatic treatment of expanded blastocysts that had been grown on schedule from frozen-thawed surplus embryos at the 2- or 4-cell stage. The blastocysts were placed on CD26-overexpressing or mock-transfected control monolayer cell cultures. The CD26-overexpression caused significantly higher blastocyst adhesion rate (53.3% versus 25.0%, P < 0.05) and significantly larger outgrowth area of trophectoderm (1.7-fold, P < 0.05). The second part of the present study was to show the expression of fibronectin, a CD26 ligand, in human preimplantation embryos, using the same donated resources. Fibronectin mRNA was detected by RT-PCR from the single hatched blastocyst (2/2) and from the single early blastocyst (3/6) but not from the single morula (0/5) samples. An indirect immunofluorescence technique verified the localization of fibronectin on the surface of the blastocyst. These results indicate that the adhesion mechanism by endometrial CD26 and embryonal fibronectin may be involved in human blastocyst implantation.     

Bufalin induces apoptosis and the G0/G1 cell cycle arrest of endometriotic stromal cells: a promising agent for the treatment of endometriosis

Most of the current medical treatments for endometriosis aim to down-regulate the estrogen activity. However, a high recurrence rate after medical treatments has been the most significant problem. Bufalin is a major digoxin-like immunoreactive component isolated from the skin and parotid venom glands of toad and is considered an apoptosis-inducing agent. To apply bufalin to the medical treatment of endometriosis, we investigated the effects of this agent on the cell proliferation and apoptosis of cultured ovarian endometriotic cyst stromal cells (ECSC) by a modified methylthiazoletetrazolium (MTT) assay, a 5-bromo-2'-deoxyuridine (BrdU) incorporation assay and internucleosomal DNA fragmentation assays. The effect of bufalin on the cell cycle of ECSC was also determined by flow cytometry. The expression of apoptosis- and cell cycle-related molecules was also examined in ECSC using Western blot analysis. Bufalin significantly inhibited the cell proliferation and DNA synthesis of ECSC and induced apoptosis and the G0/G1 phase cell cycle arrest of these cells. The down-regulation of the cyclin A, Bcl-2, and Bcl-X(L) expression with the simultaneous up-regulation of the p21 and Bax expression, and caspase-9 activation was observed in ECSC after bufalin treatment. It is suggested that bufalin induces apoptosis of ECSC by simultaneously suppressing anti-apoptotic proteins and inducing pro-apoptotic proteins. Caspase-9-mediated cascade is involved in this mechanism. Therefore, bufalin could be used as a therapeutic agent for the treatment of endometriosis.     

HeLa细胞及Ehrlich瘤细胞分别提取RA33/36抗原用于诊断RA的比较

目的比较由HeLa细胞及Ehrlich瘤细胞分别提取RA33／36抗原在类风湿性关节炎（rheumatoid arthritis，RA）诊断中的敏感性和特异性。方法RA33／36抗原由HeLa细胞及Ehrlich瘤细胞提取，抗RA33／36用免疫印迹法检测。研究对象分RA患者、非RA患者及正常对照。结果HeLa细胞抗原和Ehrlich瘤细胞抗原两者检测抗RA33／36对RA诊断的特异性为85．7％和87．5％，敏感性为28．2％和26．9％。抗RA33和抗RA36对RA均有诊断价值。结论HeLa细胞及Ehrlich瘤细胞均可用于提取RA33／36抗原。     

Hyperbaric oxygen-stimulated proliferation and growth of osteoblasts may be mediated through the FGF-2/MEK/ERK 1/2/NF-κB and PKC/JNK pathways

We investigated whether the hyperbaric oxygen (O₂) could promote the proliferation of growth-arrested osteoblasts in vitro and the mechanisms involved in this process. Osteoblasts were exposed to different combinations of saturation and pressure of O₂ and evaluated at 3 and 7 days. Control cells were cultured under ambient O₂ and normal pressure [1 atmosphere (ATA)]; high-pressure group cells were treated with high pressure (2.5 ATA) twice daily; high-O₂ group cells were treated with a high concentration O₂ (50% O₂) twice daily; and high pressure plus high-O₂ group cells were treated with high pressure (2.5 ATA) and a high concentration O₂ (50% O₂) twice daily. Hyperbaric O₂ significantly promoted osteoblast proliferation and cell cycle progression after 3 days of treatment. Hyperbaric O₂ treatment stimulated significantly increased mRNA expression of fibroblast growth factor (FGF)-2 as well as protein expression levels of Akt, p70^S6K, phosphorylated ERK, nuclear factor (NF)-κB, protein kinase C (PKC)α, and phosphorylated c-Jun N-terminal kinase (JNK). Our findings indicate that high pressure and high O₂ saturation stimulates growth-arrested osteoblasts to proliferate. These findings suggest that the proliferative effects of hyperbaric O₂ on osteoblasts may contribute to the recruitment of osteoblasts at the fracture site. The FGF-2/MEK/ERK 1/2/Akt/p70^S6K/NF-κB and PKC/JNK pathways may be involved in mediating this process.