BMP信号通路在华法林诱导的大鼠血管平滑肌细胞钙化中的作用

目的探讨骨形态发生蛋白(BMP)信号通路在华法林诱导的大鼠血管平滑肌细胞(VSMC)钙化中的作用。方法体外分离培养大鼠胸主动脉VSMC,将其随机分为正常对照组、高磷组、华法林(10μmol/L)干预组及华法林(10μmol/L)+维生素K(10μmol/L)干预组。对细胞进行钙含量和碱性磷酸酶(ALP)活性检测,茜素红染色检测钙结节,Western blot检测细胞中Runx2蛋白的表达变化,RT-PCR检测细胞中骨形态发生蛋白2(BMP-2)、Smad1及Runx2的表达变化。结果茜素红染色结果显示,与正常对照组和高磷组相比,华法林干预组钙化结节明显增多;钙含量测定结果与茜素红染色结果基本一致。与正常对照组和高磷组相比,华法林干预组ALP活性明显增加(P0.05),Runx2 mRNA和蛋白表达水平及BMP-2、Smad1 mRNA表达明显增高(P0.05);与华法林干预组相比,华法林+维生素K干预组细胞钙含量、ALP活性及BMP-2、Smad1、Runx2的表达均明显降低(P0.05)。结论BMP信号通路参与了华法林促进的大鼠VSMC钙化,其可能的作用机制是介导了VSMC的表型转化。     

Deliveries of babies with normal health derived from oocytes with smooth endoplasmic reticulum clusters

PurposeTo examine the impact on development of derived embryos from smooth endoplasmic reticulum clusters (SERC) in human metaphase II (MII) oocytes.MethodsRetrospective analysis at Kyono ART Clinic. Comparison of embryological development, pregnancy, live birth and fetal malformation between oocytes with SERC (the SERC(+) group) and those without (the SERC(−) group) in 2,158 patients (3,758 cycles) after ICSI.ResultsFertilization and implantation rate were significantly lower in SERC(+) MII oocytes than in SERC(−) MII oocytes. After the transfer of fresh and vitrified embryos derived from SERC(+) oocytes, 14 pregnancies resulted in 14 healthy babies, including 2 from fresh embryo transfer (ET) and 12 from vitrified-warmed ET, with no malformations.Conclusion(s)The presence of SERC in MII oocytes was associated with significantly lower fertilization rates and implantation rates than seen in SERC(−) MII oocytes within SERC (+) cycles. However, SERC had no impact on post-implantation development as well as neonatal outcome.     

三氧化二砷对人冠状动脉平滑肌细胞促凋亡作用的体外实验

目的 探讨三氧化二砷对人冠状动脉平滑肌细胞(HCSMC)的促凋亡作用机制及其对损伤动脉再狭窄防治的意义.方法 将人冠状动脉平滑肌细胞传代培养,将细胞分为正常对照组、三氧化二砷(浓度分别为1.0,2.0,3.0,4.0,5.0 μmol/L)组,通过原位细胞凋亡检测试剂盒(TUNEL),流式细胞仪检测细胞周期观察三氧化二砷促细胞凋亡作用,WESTERN印迹检测凋亡相关基因Bcl-2和Bax的表达变化,RT-PCR检测三氧化二砷对E2F-1的作用.结果 随三氧化二砷浓度的增加它能够抑制HCSMC的增殖并且该作用与时间及三氧化二砷浓度呈正相关(观察第7天、浓度为5.0 μmol/L时活细胞数为(4.41±0.10)×105/mL与正常对照组(30.11±0.93)×105/mL相比较差异具有统计学意义(P＜0.05),TUNEL可见凋亡细胞由16.0%±3.1%增至38.7%±2.7%(P＜0.05),流式细胞仪可见4.0 μmol/L三氧化二砷作用HCSMC 48 h后代表细胞凋亡的亚二倍体峰在48 h为最多,WESTERN印迹可见三氧化二砷上调凋亡促进基因Bax的表达而下调凋亡抑制基因Bcl-2表达.RT-PCR可见E2F-1 mRNA表达于48 h最少(P＜0.05).正常对照组均不见典型上述变化.结论 三氧化二砷可促进HCSMC的凋亡,降低HCASMC转录因子E2F-1 mRNA表达,从而抑制HCASMC的过度增殖、迁移和黏附.     

单核细胞趋化蛋白1对人脐静脉平滑肌细胞增殖的影响

目的 研究单核细胞趋化蛋白1对人脐静脉平滑肌细胞增殖的影响。方法用不同浓度单核细胞趋化蛋1（0．1、1．0、10及100μg/L）作用于人脐静脉平滑肌细胞，采用细胞计数、MTT、流式细胞术检测细胞增殖情况；采用逆转录聚合酶链反应检测细胞中c-fos基因的表达。结果单核细胞趋化蛋白1能诱导人脐静脉平滑肌细胞的增殖，呈剂量依赖关系，100μg/L单核细胞趋化蛋白1对人脐静脉血管平滑肌细胞的增殖作用强于平滑肌细胞增殖因子血小板源生长因子的作用（P〈0．001），单核细胞趋化蛋白l在诱导血管平滑肌细胞增殖的同时引起c-fos基因的高表达（P〈0．001）。结论单核细胞趋化蛋白1不仅是趋化激活剂，而且是人脐静脉平滑肌细胞的促增殖因子，并且其促增殖的功能可能与增强c-fos基因的表达有关。     

卯母细胞滑面内质网聚集的研究进展

在人类辅助生殖中,经促排卵体外获得的卵母细胞中,有滑面内质网聚集(smooth endoplasmic reticulum aggregate,SERa)形成者占有一定比例.既往研究显示,SERa对胚胎质量及妊娠结局会造成不良影响.但近年来研究显示:SERa卵母细胞可诞生完全健康的新生儿.卵母细胞胞质内SERa的形成可能与遗传因素、卵巢过度刺激、超促排卵方案及扳机日高雌二醇(E2)等多种因素相关.大多数学者认为卵母细胞SERa的形成会导致受精率、卵裂率和囊胚形成率降低及胚胎质量的下降,使得妊娠率显著降低;然而,也有多项报道已证实SERa的卵母细胞的胚胎质量与正常卵母细胞并无差异.因此对于那些临床助孕治疗中获得SERa胚胎或者胚胎数目不足的患者,可考虑移植SERa胚胎.未来,应多关注SERa对体外受精(IVF)结局的影响及着重阐明SERa形成机制,从而从根本上防止异常卵母细胞的出现.     

RhoA signaling and blood pressure: The consequence of failing to “Tone it Down”

Uncontrolled high blood pressure is a major risk factor for heart attack, stroke, and kidney failure and contributes to an estimated 25% of deaths worldwide. Despite numerous treatment options, estimates project that reasonable blood pressure (BP) control is achieved in only about half of hypertensive patients. Improvements in the detection and management of hypertension will undoubtedly be accomplished through a better understanding of the complex etiology of this disease and a more comprehensive inventory of the genes and genetic variants that influence BP regulation. Recent studies (primarily in pre-clinical models) indicate that the small GTPase RhoA and its downstream target, Rho kinase, play an important role in regulating BP homeostasis. Herein, we summarize the underlying mechanisms and highlight signaling pathways and regulators that impart tight spatial-temporal control of RhoA activity. We also discuss known allelic variations in the RhoA pathway and consider how these polymorphisms may affect genetic risk for hypertension and its clinical manifestations. Finally, we summarize the current (albeit limited) clinical data on the efficacy of targeting the RhoA pathway in hypertensive patients.     

Primary leiomyosarcoma of the thyroid gland with prior malignancy and radiotherapy:A case report and review of literature

BACKGROUND Leiomyosarcoma(LMS) of the thyroid gland is a rarely presented tumor that offers poor prognosis. To the best of the authors' knowledge, there currently exist only 28 known cases described in the literature(limited to English).CASE SUMMARY Herein a case is reported of a 60-year-old female patient who had an LMS of the thyroid, which was accompanied by periodic dysphonia and breathing disorder as well as the feeling of pressure in the chest and neck. At the time the disease was diagnosed, no metastases were detected. Prior to the diagnosis, the patient experienced a uterine adenocarcinoma that had been treated by surgical procedure and radiotherapy. For the LMS, a total thyroidectomy was performed,followed by radiotherapy. Since metastases were also discovered in the lungs,sternum, and femur, chemotherapy was administered as well.Immunohistochemically, the tumor cells in the thyroid indicated positively for alpha smooth muscle actin, calponin, and H-caldesmon, but were negative for CD34, p63, estrogen receptor, progesterone receptor, and Epstein-Barr virus.CONCLUSION Although the etiology of the LMS is as of yet unknown, prior malignancy and radiation should be considered as risk factors.     

脑源性神经营养因子对小鼠结肠平滑肌细胞的作用及其机制

目的 探讨脑源性神经营养因子(BDNF)对小鼠结肠平滑肌细胞(SMC)钙离子浓度及α-平滑肌激动蛋白(α-SMA)的影响及其相关调节机制。方法 Western blotting法检测BDNF基因敲除(BDNF^+/-)小鼠与正常野生型(BDNF^+/+)小鼠α-SMA表达水平的差异。培养原代小鼠结肠SMC,免疫荧光法检测SMC中酪氨酸激酶B(TrkB)受体的表达。同时以BDNF、TrkB受体阻滞剂(K252a)干预SMC,Western blotting法检测α-SMA和TrkB-PLC-Ca²⁺信号通路蛋白表达水平的变化,钙离子成像法检测SMC内钙离子浓度的变化。结果 与BDNF^+/+小鼠相比,BDNF^+/-小鼠结肠α-SMA表达水平明显降低。SMC表达TrkB受体,在BDNF作用下,SMC中α-SMA、TrkB-PLC-Ca²⁺信号通路蛋白表达量和细胞内钙离子浓度增加,且加入K252a可阻断以上变化。结论 BDNF可能通过TrkB-PLC-Ca²⁺信号通路作用于SMC,影响细胞内钙离子浓度及α-SMA的表达水平,进而影响肠道动力。