Craniosynostosis in Twist heterozygous mice: A model for Saethre‐Chotzen syndrome

Saethre‐Chotzen syndrome is a common autosomal dominant form of craniosynostosis, the premature fusion of the sutures of the calvarial bones of the skull. Most Saethre‐Chotzen syndrome cases are caused by haploinsufficiency for the TWIST gene. Mice heterozygous for a null mutation of the Twist gene replicate certain features of Saethre‐Chotzen syndrome, but have not been reported to exhibit craniosynostosis. We demonstrate that Twist heterozygous mice exhibit fusions of the coronal suture and other cranial suture abnormalities, indicating that Twist heterozygous mice constitute a better animal model for Saethre‐Chotzen syndrome than was previously appreciated. Anat Rec 268:90–92, 2002. © 2002 Wiley‐Liss, Inc.     

TWIST2通过调控FGF21介导的AMPK/mTOR通路诱导卵巢癌细胞氧化应激和凋亡

目的探究TWIST2对卵巢癌细胞氧化应激和凋亡的作用及相关机制。方法收集2018年6月至2019年10月在山东省妇幼保健院进行治疗的30例卵巢癌患者的癌组织和癌旁组织,将pcDNA3.1-TWIST2质粒(TWIST2组)和pcDNA3.1阴性对照质粒(阴性对照组)分别转染卵巢癌细胞(SK-OV-3和HO-8910),未转染质粒组为空白对照组。通过实时荧光定量PCR(qRT-PCR)和免疫印迹(Western blot)实验检测组织和细胞中TWIST2 mRNA和蛋白质的表达量;细胞计数试剂盒CCK-8检测TWIST2组、阴性对照组和空白对照组的细胞活力;Western blot实验检测3组的凋亡相关蛋白caspase-3、Bax和Bcl-2,AMPK/mTOR通路相关蛋白FGF21、p-AMPK和p-mTOR的蛋白表达水平;试剂盒检测3组活性氧(ROS)基因、ATP的水平和超氧化物歧化酶(SOD)的酶活性。通过检测不同处理组的细胞凋亡率和ROS水平,研究TWIST2与FGF21的相互作用。结果TWIST2在卵巢癌组织和卵巢癌细胞中的表达量显著低于癌旁组织和正常卵巢上皮细胞的表达量(P<0.05)。与空白对照组和阴性对照组相比,TWIST2组的细胞活力显著降低,细胞凋亡率显著提高(P<0.05);ROS的水平显著升高,SOD酶活力、ATP水平、NAD~+/NADH水平显著降低(P<0.05)。TWIST2可以抑制FGF21、p-AMPK、p-mTOR蛋白的表达(P<0.05);在TWIST2组中加入FGF21或AMPK激动剂时,p-AMPK、p-mTOR蛋白表达水平显著升高,卵巢癌细胞的凋亡率和ROS水平显著降低(P<0.05),TWIST2与FGF21的表达量呈负相关(r=-0.6254,P<0.01)。结论TWIST2能够促进卵巢癌细胞氧化应激和凋亡,其作用机制是通过调控FGF21介导的AMPK/mTOR通路来实现的。     

左心室旋转运动研究进展

左心室旋转运动由左心室收缩期扭转运动和舒张期解旋运动构成.左心室收缩期心内膜与心外膜下心肌纤维向相反方向运动,导致左心室扭转,进而引起左心室收缩射血.舒张期左心室心肌迅速弹性回缩释放扭转时储存的弹性势能,引起左心室早期快速充盈.任何影响心肌状态的生理或病理变化均可使左心室旋转运动发生改变.心肌收缩及松弛程度、前后负荷、心内膜及心外膜下心肌纤维收缩的平衡和心肌纤维的排列方向共同决定和影响左心室扭转运动;左心室扭转峰值、左心室弹性回缩力及心肌松弛特性决定左心室解旋运动,其他相关因素亦可影响左心室旋转运动,如运动、年龄等.该文就左心室旋转运动相关研究进展进行综述,以促进其在临床的推广应用.     

Twist、p53和E-钙黏附蛋白联合检测在乳腺癌中意义

目的 观察乳腺癌中Twist、p53和E-钙黏附蛋白mRNA的表达及其相互关系.方法 选取初次就诊的30例乳腺癌患者行改良根治术的乳腺癌组织和癌旁组织及10例乳腺良性病变患者的正常乳腺组织,分别应用RT-PCR的方法检测Twist、p53和E-钙黏附蛋白mRNA的表达,与临床病理因素关系及其相互关系.结果 乳腺癌组织Twist mRNA表达与分化程度[高中分化93.33％(14/15),低分化53.33％(8/15)]、淋巴管浸润[是93.75％(15/16),否50.00％(7/14)]和TNM分期[Ⅰ～Ⅱ期46.15％(6/13),III～Ⅳ期94.12％(16/17)]有明显相关性(P＜0.05),而与年龄、肿瘤直径、外膜浸润、雌激素受体和孕激素受体无明显相关性(P＞0.05).乳腺癌组织中Twist和p53 mRNA表达较癌旁组织和正常乳腺组织明显升高[(112.36±12.53)％比(101.37±9.12)％和(98.03±9.04)％、(101.73±13.32)％比(91.43±7.43)％和(90.83±6.58)％,P＜0.01],而E-钙黏附蛋白mRNA表达较癌旁组织和正常乳腺组织明显降低[(55.43±12.49)％比(69.39±5.63)％和(72.73±7.93)％,P＜0.01],癌旁组织和正常乳腺组织比较差异无统计学意义(P＞0.05).乳腺癌组织中Twist mRNA表达与p53 mRNA呈正相关(r=0.526,P=0.004),而与E-钙黏附蛋白mRNA表达呈负相关(r=-0.500,P=0.000).结论 Twist和p53在乳腺癌组织中呈高表达,而E-钙黏附蛋白呈低表达.Twist可能通过上调p53的表达,同时下调E-钙黏附蛋白的表达,从而促进乳腺癌的浸润、转移,并在其中起重要作用.     

不锈钢麻花牵引圈的制作及在管腔器械清洗质量检查中的应用

目的 制作不锈钢麻花牵引圈并探讨其在管腔器械清洗质量检查中的应用效果。方法 将制作的不锈钢麻花牵引圈用于管腔器械清洗质量的检查,通过与目测+光源放大镜法、探条纱布法、ATP生物荧光法做比较,评价其在管腔器械清洗质量检查中的效果。结果 A类空腔器械清洗质量检查中,麻花牵引圈法、目测+放大镜法和ATP生物荧光法的清洗质量检查合格率差别有统计学意义（P<0.010）,两两比较,目测+放大镜法合格率最高,与其他2组的检查合格率差异具有统计学意义（P<0.017）,麻花牵引圈法与ATP生物荧光法的检查合格率差异无统计学意义（P>0.017）。B类空腔器械清洗质量检查中,4种清洗质量检查方法合格率差异具有统计学意义（P<0.010）,两两比较,目测+放大镜法合格率最高,与其他3组的检查合格率差异具有统计学意义（P<0.008）。麻花牵引圈法、探条纱布法和ATP生物荧光法的检查合格率差异无统计学意义（P>0.008）。结论 不锈钢麻花牵引圈在管腔器械清洗质量检查中,能发现目测+放大镜法无法检出的不合格器械,其检测效果与探条纱布法、ATP生物荧光法相当,比ATP生物荧光法节省费用,但操作较金属探条法便捷,适用范围更广。     

Twist基因逆转录病毒载体的构建及其诱导正常乳腺上皮细胞发生上皮间质转化

目的 构建Twist基因逆转录病毒载体,研究其对人正常乳腺上皮细胞MCF10A的影响.方法 酶切pcDNA3/myc-Twist获得myc-Twist,克隆到逆转录病毒载体pBABE-puro中,构建重组质粒pB AB E-myc-Twist.通过酶切、测序鉴定Twist基因正确后,在体外将质粒pBABE-myc-Twist和其对照分别与包装质粒pAmpho共同转染人胚肾上皮细胞系293T细胞,进行逆转录病毒的包装,并感染人正常乳腺MCF10A上皮细胞,用嘌呤霉素(puromycin)筛选,获得成功转入Twist的MCF10A-Twist细胞和MCF10A-Vector对照细胞.通过RT-PCR和Western blot法验证Twist细胞在MCF10A-Twist和MCF10A-Vector细胞内的表达.免疫荧光技术和Western blot法检测细胞内上皮间质转化(EMT)标志蛋白的表达.Transwell(R)法检测细胞的迁移和侵袭能力.结果 重组逆转录病毒载体质粒pBABE-myc-Twist经酶切和测序鉴定构建正确;Twist基因被逆转录病毒成功导入MCF10A细胞,并在靶细胞内稳定表达,RT-PCR检测到Twist mRNA在MCF10A-Twist细胞中表达,Western blot法检测发现myc-Twist蛋白在MCF10A-Twist细胞中表达;免疫荧光和Western blot法检测显示在MCF10A-Twist细胞中E-cadherin表达显著下调、vimentin表达明显上调;与MCF10A-Vector细胞相比,Transwell(R)法检测发现MCF10A-Twist细胞迁移和侵袭能力明显增加(P＜0.01).结论 Twist诱导MCF10A细胞发生EMT转化,并促进其迁移和侵袭.     

miR-580对乳腺癌细胞系中Twist1的调节作用

目的：探讨miR-580在MCF-10A细胞系中对Twist1的调节。方法：本实验利用生物信息学方法预测Twist1的靶miRNA是miR-580。首先,采用qPCR法检测在MCF-10A系列细胞系中Twist1及miR-580的表达。然后,在MCF-10A细胞系中分别转染miR-580类似物和miR-580抑制物后,利用RT-PCR、Western blot、t检验分析Twist1的表达及细胞迁移能力的变化。最后利用荧光素酶实验验证miR-580通过结合在Twist1的3'UTR调节其表达。结果：1)在MCF-10A细胞系中Twist1与miR-580的表达呈负相关;2)在MCF-10A细胞系中转染miR-580类似物后,Twist1的表达下调;在MCF-10A细胞系中转染miR-580抑制物后Twist1的表达上调;3)在MCF-10A细胞系中引入miR-580类似物后细胞迁移能力降低;4)miR-580直接结合在Twist1的3'UTR。结论：miRNA-580在MCF-10A细胞系中通过结合在Twist1的3'UTR负向调节Twist1的表达从而克制细胞的迁移。     

Effects of Boiled Dill Seed on Anxiety During Labor: A Randomized Controlled Trial

Objective: To evaluate the effect of boiled dill seed (Anethum graveolens) on anxiety during childbirth compared with oxytocin. Methods: This study was a randomized controlled trial with 100 pregnant women who were assigned to the oxytocin (50 cases) and the boiled dill seed (50 cases) group by a table of random number. In the boiled dill seed group, 10 g of dill seed was boiled in 100 mL for 10 min and was given to women after filtration once orally at the beginning of active phase. In the oxytocin group, 10 IU of oxytocin in 1000 mL of Ringer solution was prescribed according to the clinical routine. The State-Trait Anxiety Inventory (STAI) in both groups before (at the beginning of the active phase) and after (6 h after the delivery) was completed. Meanwhile, pain intensity and duration of labor, dilatation and effacement scores, Apgar score of newborns, and FHR were measured and evaluated. Results: No statistically significant differences were shown in obvious anxiety scores between the two groups at baseline (P>0.05). After the delivery, in the boiled dill seed group, the number of women with severe [0 vs. 8.0% (4/50)] and almost severe [0 vs. 14.0% (7/50)] trait and almost severe [0 vs. 14.0% (7/50)] state anxiety was lower than those in the control group (P=0.050, P=0.041, respectively). Moreover, labor was shorter in the 1st (P<0.01), 2nd (P=0.78) and 3rd (P=0.10) stages in the boiled dill seed group compared to the control group. Conclusions: Dill seed could be used as an effective treatment to reduce anxiety during labor. Dill seed can be effective in reducing the length of labor. Due to lack of maternal and fetal complications, the boiled dill seed could also be used to reduce cesarean section rates in women who are fearful and anxious of delivery. (RCT Code: IRCT201607177065N2)