高血压病人小动脉中磷酸化细胞外信号调节激酶1/2及ets样基因1的蛋白表达

目的研究细胞外信号调节激酶系统（ERK）及其下游底物ets样基因1（E1k-1）在原发性高血压中的作用。方法采用免疫组织化学方法，对比观察高血压和非高血压病人胃肠小动脉血管平滑肌细胞及内皮细胞中细胞外信号调节激酶（ERK1／2）和ets样基因1（E1k-1）的磷酸化情况。结果高血压组胃肠小动脉血管平滑肌细胞中磷酸化ERK1／2染色阳性率（8．31％）明显高于非高血压组（0．53％），P〈0．05；高血压组内皮细胞中磷酸化ERK1／2的阳性率（4．97％）明显高于非高血压组（P〈0．05）。高血压组胃肠小动脉血管平滑肌细胞中磷酸化Elk-1染色阳性率（3．53％）明显高于非高血压组（0．27％），P〈0．05。在血管平滑肌细胞、血管内皮细胞中磷酸化ERK1／2与磷酸化E1k-1的表达均有正相关关系。结论原发性高血压患者的胃肠细小动脉血管平滑肌细胞以及内皮细胞中细胞外信号调节激酶及其下游底物Elk-1的磷酸化增加。     

高血压大鼠血管紧张素Ⅱ对血管平滑肌细胞PAI-1表达的作用与ERK及AT1受体的关系

目的观察高血压大鼠的血管内皮细胞和平滑肌细胞上胞外信号调节激酶(ERK)和血管紧张素Ⅱ(Ang Ⅱ)1型受体(AT1R)的变化与Ang Ⅱ对纤溶酶原激活物抑制物-1(PAI-1)活性调节作用的内在因果关系,探讨AngⅡ促血管内皮细胞和血管平滑肌细胞合成与分泌PAI-1的受体和受体后信号途径.方法将16只健康SD大鼠随机分为腹主动脉缩窄型高血压组(n=8)和假手术对照组(n=8).第6周时应用放射免疫法检测大鼠血浆与主动脉组织匀浆中Ang Ⅱ含量,发色底物法检测血浆与主动脉孵育液中PAI-1活性的变化,免疫组织化学法测定ERK和AT1R在血管内皮细胞及血管平滑肌细胞的表达.结果血浆Ang Ⅱ、血浆PAI-1、血管平滑肌细胞ERK、血管平滑肌细胞AT1R均显著增高(P均＜0.05),且四者之间互为正相关(r=0.89～0.96,P＜0.01～0.001),主动脉匀浆Ang Ⅱ、主动脉孵育液PAI-1、血管平滑肌细胞ERK、血管平滑肌细胞AT1R也显著增高(P均＜0.05),且四者之间亦互为正相关(r=0.86～0.96, P＜0.01～0.001).结论高血压时Ang Ⅱ具有诱导PAI-1合成与分泌的作用,可能与Ang Ⅱ经AT1R激活ERK,介导血管平滑肌合成及释放PAI-1有关.     

替米沙坦对高血压大鼠纤溶功能的作用机制

目的 探讨血管紧张素Ⅱ(Ang Ⅱ)促血管内皮细胞和平滑肌细胞合成与分泌纤溶酶原激活物抑制物-1(PAI-1)的受体和受体后信号转导途径以及替米沙坦对高血压大鼠纤溶参数的影响和可能机制.方法 腹主动脉部分缩窄构建高血压大鼠模型,随机分成高血压组和替米沙坦组(3 mg/kg·d)6周,另设假手术组为对照组.检测大鼠血浆与主动脉组织匀浆中Ang Ⅱ含量,血浆与主动脉孵育液中纤溶酶原激活物(t-PA)、PAI-1活性,血管内皮细胞、平滑肌细胞胞外信号调节激酶(ERK)和血管紧张素Ⅱ1型受体(AT1R)蛋白的表达.结果 ①高血压组血浆Ang Ⅱ含量、血浆PAI-1活性、血管平滑肌细胞ERK蛋白及AT1 R蛋白的表达4者之间显著正相关(r=0.89～0.96,P＜0.01～0.001).主动脉匀浆Ang Ⅱ含量、主动脉孵育液PAI-1活性、血管平滑肌细胞ERK蛋白及AT1R蛋白的表达4者之间显著正相关(r=0.86～0.96,P＜0.01～0.001).②与高血压组比较,替米沙坦能明显抑制主动脉分泌PAI-1的能力(P＜0.05),降低血浆PAI-1活性(P＜0.05),升高血浆与主动脉孵育液中t-PA活性、t-PA/PAI-1值(P均＜0.05),明显改善纤溶参数.③替米沙坦组ERK和AT1R在血管内皮细胞和平滑肌细胞的表达较高血压组明显减少(P均＜0.05).结论 替米沙坦抑制血管内皮细胞、血管平滑肌细胞ERK和AT1 R的表达,抑制Ang Ⅱ引起的PAI-1合成增加,可能是其改善纤溶功能的机制.     

高血压大鼠血管紧张素Ⅱ对血管平滑肌细胞PAI-1表达的作用与ERK及AT1受体的关系

目的　观察高血压大鼠的血管内皮细胞和平滑肌细胞上胞外信号调节激酶 (ERK)和血管紧张素Ⅱ (AngⅡ ) 1型受体 (AT1R)的变化与AngⅡ对纤溶酶原激活物抑制物 - 1(PAI 1)活性调节作用的内在因果关系 ,探讨AngⅡ促血管内皮细胞和血管平滑肌细胞合成与分泌PAI 1的受体和受体后信号途径。方法　将 16只健康SD大鼠随机分为腹主动脉缩窄型高血压组 (n =8)和假手术对照组 (n =8)。第 6周时应用放射免疫法检测大鼠血浆与主动脉组织匀浆中AngⅡ含量 ,发色底物法检测血浆与主动脉孵育液中PAI 1活性的变化 ,免疫组织化学法测定ERK和AT1R在血管内皮细胞及血管平滑肌细胞的表达。结果　血浆AngⅡ、血浆PAI 1、血管平滑肌细胞ERK、血管平滑肌细胞AT1R均显著增高 (P均 <0 0 5 ) ,且四者之间互为正相关 (r =0 89～ 0 96 ,P <0 0 1～ 0 0 0 1) ,主动脉匀浆AngⅡ、主动脉孵育液PAI 1、血管平滑肌细胞ERK、血管平滑肌细胞AT1R也显著增高 (P均 <0 0 5 ) ,且四者之间亦互为正相关 (r =0 86～ 0 96 ,P <0 0 1～ 0 0 0 1)。结论　高血压时AngⅡ具有诱导PAI 1合成与分泌的作用 ,可能与AngⅡ经AT1R激活ERK ,介导血管平滑肌合成及释放PAI 1有关。     

替米沙坦对高血压大鼠纤溶功能的作用机制

目的探讨血管紧张素Ⅱ(AngⅡ)促血管内皮细胞和平滑肌细胞合成与分泌纤溶酶原激活物抑制物1(PAI1)的受体和受体后信号转导途径以及替米沙坦对高血压大鼠纤溶参数的影响和可能机制。方法腹主动脉部分缩窄构建高血压大鼠模型,随机分成高血压组和替米沙坦组(3mg/·d)6周,另设假手术组为对照组。检测大鼠血浆与主动脉组织匀浆中AngⅡ含量,血浆与主动脉孵育液中纤溶酶原激活物(tPA)、PAI1活性,血管内皮细胞、平滑肌细胞胞外信号调节激酶(ERK)和血管紧张素Ⅱ1型受体(AT1R)蛋白的表达。结果①高血压组血浆AngⅡ含量、血浆PAI1活性、血管平滑肌细胞ERK蛋白及AT1R蛋白的表达4者之间显著正相关(r=0.89～0.96,P<0.01～0.001)。主动脉匀浆AngⅡ含量、主动脉孵育液PAI1活性、血管平滑肌细胞ERK蛋白及AT1R蛋白的表达4者之间显著正相关(r=0.86～0.96,P<0.01～0.001)。②与高血压组比较,替米沙坦能明显抑制主动脉分泌PAI1的能力(P<0.05),降低血浆PAI1活性(P<0.05),升高血浆与主动脉孵育液中tPA活性、tPA/PAI1值(P均<0.05),明显改善纤溶参数。③替米沙坦组ERK和AT1R在血管内皮细胞和平滑肌细胞的表达较高血压组明显减少(P均<0.05)。结论替米沙坦抑制血管内皮细胞、血管平滑肌细胞ERK和AT1R的表达,抑制AngⅡ引起的PAI1合成增加,可能是其改善纤溶功能的机制。     

高血压大鼠血管平滑肌细胞中ERK-2和原癌基因c-jun的表达

目的ERK-2是细胞外信号调节激酶,本文研究MAPK/ERK系统在高血压中的作用及其机制.方法Wistar大鼠两肾一夹型高血压模型,4周后处死,用免疫组织化学方法,观察肾细小动脉平滑肌细胞中ERK-2和c-jun表达.结果实验期间,高血压组动脉血压从(112±18)mmHg升高到实验结束时的(198±33)mmHg;高血压组肾小球发生了明显的纤维化(P＜0.05);高血压组可见肾入球动脉玻璃样变,小动脉发生了管壁增厚、细胞数目增多;高血压组肾小叶间动脉ERK-2染色阳性率(16.86%)高于对照组(P＜0.01),ERK-2在高血压组入球动脉、小叶间动脉、叶间动脉和弓形动脉VSMC中染色阳性率(分别为7.09%、14.57%,29.44%及13.63%)均明显高于对照组(P＜0.01);在高血压组入球动脉及其VSMC中c-jun的阳性率(分别为17.62%和8.73%)明显高于对照组(P＜0.01),小叶间动脉、弓形动脉VSMC中c-jun染色阳性率均明显高于对照组(P＜0.01).结论两肾一夹型高血压时,可出现ERK-2过表达,转录相关基因c-jun活化,最终使VSMC增多.     

胰岛素样生长因子-1对大鼠结肠平滑肌细胞增殖、凋亡及MAPK通路的影响

目的 探讨胰岛素样生长因子-1(IGF-1)对大鼠结肠平滑肌细胞(SMCs)增殖、凋亡及丝裂原活化蛋白激酶(MAPK)信号转导通路的影响.方法 利用酶解法分离培养Sprague-Dawley大鼠结肠SMCs,进行免疫组化染色鉴定后,将其分为对照组、IGF-1组和IGF-1+ PD98059[细胞外信号调节激酶(ERK)抑制剂]组,分别采用噻唑蓝(MTT)法检测SMCs增殖,流式细胞术AnnexinV-FITC/PI检测SMCs凋亡,Western印迹检测磷酸化ERK、ERK、磷酸化p38MAPK、p38MAPK和磷酸化c-Jun氨基末端激酶(JNK)、JNK的表达.结果 分离培养的细胞经免疫组化鉴定为结肠SMCs,IGF-1组较对照组细胞增殖增强[(1.786 ±0.271)比(0.998±0.057),P＜0.01],凋亡率降低[(2.59±0.28)％比(20.68±2.48)％,P＜0.01],磷酸化ERK表达增强,磷酸化ERK/ERK比值升高[(42.71±3.74)％比(23.88±2.52％),P均＜0.01];磷酸化p38MAPK、p38MAPK、磷酸化JNK、JNK表达无差异(P均＞0.05).IGF-1+ PD98059组较对照组细胞增殖下降[(0.154±0.021)比(0.998±0.057),P＜0.01],凋亡率升高[(84.31±7.54)％比(20.68±2.48)％,P＜0.01],磷酸化ERK表达减弱,磷酸化ERK/ERK比值降低[(10.47±1.22)％比(23.88±2.52)％,P均＜0.01].结论 IGF-1可能通过激活结肠SMCs MAPK通路中的ERK途径,促进细胞增殖,抑制凋亡,可能与p38MAPK途径和JNK途径无关.     

KATP通道与人肺动脉平滑肌细胞外信号调节激酶磷酸化的关系

目的 研究三磷酸腺苷(ATP)敏感性钾(KATP)通道与人肺动脉平滑肌细胞外信号调节激酶1和2(ERK1/2)磷酸化的关系.方法 原代培养人肺动脉平滑肌细胞,用Western-blot方法 检测磷酸化细胞外信号调节激酶1和2(p-ERK1/2).对照组不予干预,实验组分别加入内皮素-1(ET-1)、ET-1+埃他卡林、吡那地尔或格列本脲等孵育.结果 ①在2～30 min,ET-1呈时间依赖性促进人肺动脉平滑肌细胞ERK1/2磷酸化,10 min时最明显.②埃他卡林和吡那地尔可拮抗ET-1对ERK1/2磷酸化的影响.③特异性KATP通道阻断剂格列本脲可逆转埃他卡林和吡那地尔的作用.结论 KATP通道开放剂可能通过激活KATP通道,抑制ET-1诱导的原代培养人肺动脉平滑肌细胞ERK1/2磷酸化,KATP通道可能是研发新型治疗肺动脉高压药物的重要靶分子.     

血管紧张素(1-7)阻断细胞外信号调节激酶1/2、核因子κB信号通路影响泡沫细胞内胆固醇含量

目的研究血管紧张素(1-7)对THP-1源性泡沫细胞中细胞外信号调节激酶1/2及核因子κB信号转导通路的影响,以进一步探讨血管紧张素(1-7)促进胆固醇逆转运的调节机制以及细胞外信号调节激酶1/2与核因子κB信号通路之间是否相互影响。方法采用体外培养的THP-1单核细胞构建泡沫细胞模型,用不同的干预方法处理细胞72 h,将细胞分为单核细胞(空白对照)组、泡沫细胞组、分别预先经10-6mol/L血管紧张素(1-7)、10μmol/L核因子κB特异性阻断剂、10μmol/L细胞外信号调节激酶1/2特异性阻断剂、10-6mol/L血管紧张素(1-7)+10μmol/L核因子κB特异性阻断剂、10-6mol/L血管紧张素(1-7)+10μmol/L细胞外信号调节激酶1/2特异性阻断剂干预的泡沫细胞组。油红O染色后显微镜下观察细胞形态;高效液相色谱法检测细胞内胆固醇含量的变化;免疫组化法检测细胞内核因子κB(p65)活性的表达;免疫印迹法检测磷酸化细胞外信号调节激酶1/2蛋白的表达。结果血管紧张素(1-7)显著降低了泡沫细胞内胆固醇的含量,下调了磷酸化细胞外信号调节激酶1/2、核因子κB(p65)活性的表达(P0.05),细胞外信号调节激酶1/2、核因子κB信号通路被特异性阻断后泡沫细胞内胆固醇含量降低(P0.05);血管紧张素(1-7)联用细胞外信号调节激酶1/2、核因子κB信号通路的特异性阻断剂后泡沫细胞内胆固醇含量显著降低(P0.01);核因子κB信号通路被阻断后核因子κB(p65)活性表达显著降低(P0.01),而磷酸化细胞外信号调节激酶1/2活性表达无明显降低(P0.05),细胞外信号调节激酶1/2信号通路被阻断后磷酸化细胞外信号调节激酶1/2和核因子κB(p65)活性表达均降低(P0.05)。结论血管紧张素(1-7)可能通过降低磷酸化细胞外信号调节激酶1/2、核因子κB(p65)的活性,减少细胞内胆固醇的蓄积;细胞外信号调节激酶1/2、核因子κB信号通路被特异性阻断后可减少泡沫细胞内胆固醇的含量;细胞外信号调节激酶可能是核因子κB(p65)信号通路的上游信号。     

氧化型低密度脂蛋白对血管平滑肌细胞小凹蛋白1表达的抑制作用及其与信号调节激酶的关系

目的 观察氧化型低密度脂蛋白对血管平滑肌细胞小凹蛋白1表达的抑制作用及其与细胞外信号调节激酶活性的关系.方法 50 mg/L 氧化型低密度脂蛋白处理血管平滑肌细胞不同时间,或同时加入25 μmol/L细胞外信号调节激酶信号通路的特异性阻断剂,Western 印迹检测血管平滑肌细胞小凹蛋白1和细胞外信号调节激酶的变化.结果 50 mg/L氧化型低密度脂蛋白作用细胞24 h后小凹蛋白1的表达明显下降,细胞外信号调节激酶磷酸化活性显著增高,阻断氧化型低密度脂蛋白对细胞外信号调节激酶的激活可显著促进小凹蛋白1的表达.结论 血管平滑肌细胞源性泡沫细胞小凹蛋白1的表达下调与氧化型低密度脂蛋白激活细胞外信号调节激酶及其介导的信号传导密切相关.     

MEK1/2-ERK1/2 mediates alpha1-adrenergic receptor-stimulated hypertrophy in adult rat ventricular myocytes

We examined the relative roles of the mitogen-activated protein kinases (MAPK) in mediating the alpha1-adrenergic receptor (alpha1-AR) stimulated hypertrophic phenotype in adult rat ventricular myocytes (ARVM). Norepinephrine (NE; 1 microM) in the presence of the beta -AR antagonist propranolol (Pro; 2 microM) caused activation of Ras (>six-fold), MAPK/ERK kinase 1 and 2 (MEK1/2, >10-fold) and extracellular signal-regulated kinases 1 and 2 (ERK1/2, approximately 30-fold) within 5 min, as determined by kinase activity assays and Western blots using phospho-specific antibodies. Conversely, p38 and c-Jun amino-terminal kinases (JNK) were not activated by NE/Pro. Activated MEK1/2 signals remained detectable at 2 h, and activated ERK1/2 remained detectable at 48 h. The alpha1-AR selective inhibitor prazosin (100 nM) completely inhibited the NE/Pro-stimulated activation of Ras, MEK1/2 and ERK1/2. The MEK inhibitor PD98059 caused a concentration-dependent inhibition of NE/Pro-stimulated protein synthesis (as assessed by [3H]leucine incorporation and cellular protein accumulation) and ERK1/2 activation, with approximately 50% inhibition at a concentration between 10 and 50 microM, which is consistent with the known IC50 values of PD98059 for MEK1 (4 microM) and MEK2 (50 microM). Thus, these data show that alpha1-AR stimulated hypertrophy in ARVM is dependent on the MEK1/2-ERK1/2 signaling pathway.     

Cytoplasmic FMRP interacting protein 1/2 (CYFIP1/2) expression analysis in autism

Metabolic Brain Disease - Cytoplasmic FMRP interacting proteins 1 and 2 (CYFIP1/2) have been previously shown to be associated with central nervous system (CNS) disorders such as autism spectrum...     

Switchable S = 1/2 and J = 1/2 Rashba bands in ferroelectric halide perovskites

The Rashba effect is spin degeneracy lift originated from spin–orbit coupling under inversion symmetry breaking and has been intensively studied for spintronics applications. However, easily implementable methods and corresponding materials for directional controls of Rashba splitting are still lacking. Here, we propose organic–inorganic hybrid metal halide perovskites as 3D Rashba systems driven by bulk ferroelectricity. In these materials, it is shown that the helical direction of the angular momentum texture in the Rashba band can be controlled by external electric fields via ferroelectric switching. Our tight-binding analysis and first-principles calculations indicate that and Rashba bands directly coupled to ferroelectric polarization emerge at the valence and conduction band edges, respectively. The coexistence of two contrasting Rashba bands having different compositions of the spin and orbital angular momentum is a distinctive feature of these materials. With recent experimental evidence for the ferroelectric response, the halide perovskites will be, to our knowledge, the first practical realization of the ferroelectric-coupled Rashba effect, suggesting novel applications to spintronic devices.The Rashba effect has been widely investigated in 2D surfaces, interfaces, quantum wells, and 3D bulk systems (1–6). The essential requisite for the Rashba effect is that the spin degeneracy is lifted by the inversion symmetry-breaking (ISB) field in the presence of the spin–orbit coupling (SOC). To date, major concerns have focused on enlarging Rashba strength characterized by the Rashba coefficient (3, 5, 6). The controllability in the direction of the ISB field, on the other hand, has not been seriously considered. In a surface or an interface, the potential gradient generated by the structural inversion asymmetry results in the loss of controllability; the field direction is mainly fixed according to the preformed surface or interface configuration. The situation is similar in recently discovered 3D Rashba material BiTeI (6), because this material has the compositional ISB field between Te and I layers.Controlling the ISB field and ultimately Rashba-type band splitting can be achieved by using a novel ferroelectric Rashba material (7, 8). In a ferroelectric system, the bulk polarization controlled by external electric fields generates the ISB field. Therefore, the ferroelectric polarization directly couples to the spin splitting and the helical spin texture in the ferroelectric Rashba material, enabling the helicity reversal via the ferroelectric switching. Recent theoretical study suggested GeTe as a possible candidate for this mechanism, but the direct measurement of the ferroelectric polarization and switching is still missing due to the sizable conductivity of bulk GeTe (7, 8).We consider organic–inorganic hybrid metal halide perovskites as promising ferroelectric Rashba materials. The general formula for this material class is where , i.e., methylammonium (MA); M = Pb and Sn; and X = I and Br. The materials have several advantages over GeTe. Firstly, polar distortions and ferroelectric responses in the halide perovskite have been clearly observed in experiments (9, 10). On the other hand, GeTe inevitably generates Ge vacancies to give considerable bulk conductivity, which in turn hinders the polarization switching (11). Secondly, unlike the multiple band edges and strong hexagonal warping in GeTe (7, 8), the band edge states of the halide perovskites lie at a single point in the Brillouin zone with nearly isotropic Rashba bands. Thus, the halide perovskites have ideal Rashba-split bands with proper material quality. Finally, whereas GeTe has an indirect band gap (11), the halide perovskites have a direct one. This direct gap becomes important when we consider the transition between the valence and conduction bands in optical devices.Interestingly, the above halide perovskite series has two contrasting types of Rashba bands simultaneously: the and Rashba bands at the valence and conduction bands, respectively (Fig. 1). This originates from the different band characters in terms of the angular momentum at the valance and conduction band edges common in halide perovskite compounds (12). The angular momentum character of the individual band can be affected by relative energy scales of the crystal field and SOC; the crystal field quenches orbital degrees of freedom, whereas SOC entangles spin and orbital degrees. Through the competition between the two energy scales, the band character on which the Rashba Hamiltonian is based can vary from the fully spin–orbital entangled total angular momentum state (J) to the spin state (S) (13), which causes a significant distinction in the angular momentum texture of the Rashba band.Open in a separate windowFig. 1.Schematic illustration of the switchable Rashba effect. The polarization-coupled Rashba-type band splitting of the and manifolds in hybrid metal halide perovskites.In this work, we examine the electronic structures of the hybrid metal halide perovskites as candidates for the ferroelectric Rashba materials. By constructing a minimal tight-binding (TB) model Hamiltonian, we can capture the key features of the band structures; the low-energy effective Hamiltonian gives rise to the ferroelectric-coupled and Rashba bands at the valence band maximum (VBM) and the conduction band minimum (CBM), respectively. The Rashba-type splitting of the fully spin–orbital-entangled subspace stems from the bandgap-independent intraorbital as well as the bandgap-dependent interorbital terms. On the contrary, Rashba splitting only consists of the interorbital term (14). We present several examples of possible ferroelectric Rashba materials (β-MAPbI₃, β-MASnI₃, and ortho-MASbBr₃) by adopting first-principles electronic structure calculations based on density functional theory. These halide perovskites are shown to have the characteristic features predicted in the TB model with the sizable Rashba coefficient . A different type of controllability on the relative helicity between the two Rashba bands is also discussed according to the positions of the lateral halide atoms.     

A1/A2-Untergruppenbestimmungen in Abstammungsprozessen

Zusammenfassung Verfasser gibt die Resultate seiner in 1460 Rechtsfällen an 1500 vermutlichen Vätern unternommenen A₁/A₂-Untergruppen-Untersuchungen bekannt. In 15 Fällen, bzw. bei 16 Männern (d. h. 1,06% der Männer) gelang es, die Vaterschaft mit Hilfe der Untergruppenbestimmungen auszuschließen. Die absolure Zahl der Vaterschaftsausschließungen kann somit um 0,466%, erhöht werden. Der Verfasser vertritt den Standpunkt, daß, falls die Forderungen streng eingehalten werden, dem Vaterschaftsausschluß auf Grunde der Untergruppenbefunden volle Beweiskraft zuzuschreiben ist.

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Summary The author presents the results of his A₁/A₂ subgroup examinations made in 1460 court cases in 1500 assumed fathers. In 15 cases or in 16 men (i.e. 1.06% of the men) it was possible to exclude fatherhood by means of the subgroup determinations. The absolute number of fatherhood-exclusions thus may be increased by 0.466%. The author holds the view that the requirements should be kept a strict secret, in order to ensure full conclusiveness to the exclusion of fatherhood on the basis of subgroup findings.

     

Ultraviolet-A1 phototherapy modulates Th1/Th2 and Tc1/Tc2 balance in patients with systemic lupus erythematosus

OBJECTIVE: Ultraviolet-A1 (UVA1) phototherapy is effective for a variety of dermatological diseases. We examined the effectiveness and reliability of low-dose UVA1 phototherapy (60 kJ/m2/treatment) in patients suffering from systemic lupus erythematosus (SLE). We studied the changes in immunological parameters. METHODS: The patients received a 9-week course of phototherapy according to the following regimen: five times a week during the first 3 weeks, three times a week during the second 3 weeks and twice during the last 3 weeks. Among other things, we analysed the proportions of T helper 1 (Th1), Th2, T cytotoxic (Tc1) and Tc2 cell populations in the peripheral blood of patients by flow cytometric detection of intracytoplasmic interferon gamma (IFN-gamma) and interleukin 4 (IL-4). RESULTS: Our study showed the improvement of clinical symptoms determined by the subjective clinical disease activity scoring and the SLE Disease Activity Index (SLEDAI). By the end of UVA1 phototherapy, the mean value of SLEDAI had decreased from 7.2+/-5.6 to 0.9+/-1.8, which was significant (P = 0.005). Immunological investigations detected a decrease in the frequency of IFN-gamma-producing Th1 and Tc1 cells and a decrease in the Th1/Th2 and Tc1/Tc2 ratios after UVA1 therapy. CONCLUSION: According to the literature, IFN-gamma has a pathogenic role in the development of SLE. We observed a decreased proportion of IFN-gamma-secreting cells, which we think is presumably one of the beneficial effects of UVA1 therapy. On the basis of our study, UVA1 phototherapy does seem to be an effective adjuvant in the treatment of SLE patients.     

Epitaxial Growth of Diamond-Shaped Au1/2Ag1/2CN Nanocrystals on Graphene

Epitaxial synthesis of inorganic nanomaterials on pristine 2D materials is of interest in the development of nanostructured devices and nanocomposite materials, but is quite difficult because pristine surfaces of 2D materials are chemically inert. Previous studies found a few exceptions including AuCN, AgCN, CuCN, and Cu_0.5Au_0.5CN, which can be preferentially synthesized and epitaxially aligned onto various 2D materials. Here, we discover that Au_1/2Ag_1/2CN forms diamond-shaped nanocrystals epitaxially grown on pristine graphene surfaces. The nanocrystals synthesized by a simple drop-casting method are crystallographically aligned to lattice structures of the underlying graphene. Our experimental investigations on 3D structures and the synthesis conditions of the nanocrystals imply that the rhombic 2D geometries originate from different growth rates depending on orientations along and perpendicular to 1D molecular chains of Au_1/2Ag_1/2CN. We also perform in situ TEM observations showing that Au_1/2Ag_1/2CN nanocrystals are decomposed to Au and Ag alloy nanocrystals under electron beam irradiation. Our experimental results provide an additional example of 1D cyanide chain families that form ordered nanocrystals epitaxially aligned on 2D materials, and reveal basic physical characteristics of this rarely investigated nanomaterial.     

IL-1 RN 2/2 genotype and simultaneous carriage of genotypes IL-1 RN 2/2 and IL-1beta-511 T/T associated with oesophagitis in Helicobacter pylori-negative patients

BACKGROUND: Interleukin-1 receptor antagonist genotype 2/2 is associated with a prolonged and enhanced inflammatory response. It is suspected of being a risk factor for atrophic gastritis and gastric cancer and for some autoimmune diseases. No specific genetic risk factors for oesophagitis have been identified so far and there are no reports of IL-1 polymorphism in relation to oesophageal disease. METHODS: We studied the IL-1RN, IL-1beta-511 and IL-1beta + 3953 polymorphisms in an unselected series of 142 adult patients scheduled for gastrointestinal endoscopy because of dyspepsia. The control group consisted of university staff and students (n = 179). Helicobacter pylori status was determined by antibody testing and bacterial detection. RESULTS: Endoscopic oesophagitis was noted in 40 patients. The IL-1RN 2/2 genotype was significantly more prevalent in the patients with H. pylori-negative oesophagitis than in the control subjects (27% versus 9%; OR 3.574, CI 1.23-10.35, P = 0.034) or in the dyspeptic patients (27% versus 7%; OR 5.089. CI 1.51-17.11, P = 0.009). IL-1beta-511 T/T genotype tended to be more frequent in the H. pylori-negative patients with oesophagitis than in the control subjects (P = 0.071). The strongest association was between the simultaneous carriage of genotypes IL-1RN 2/2 and IL-1beta -511 T/T and H. pylori-negative oesophagitis. where the combined genotype was more prevalent than in the control subjects (23% versus 6%; OR 4.492, CI 1.40-14.46, P = 0.012) or the dyspeptic patients without oesophagitis (23% versus 3%: OR 9.706. CI 2.12-44.42, P = 0.003). CONCLUSIONS: The results indicate that the IL-1RN 2/2 genotype and the carriage of combined genotypes IL-1RN 2/2 + IL-1beta-511 T/T are associated with H. pylori-negative oesophagitis. This is the first report on the association between IL-1 gene polymorphism and oesophagitis.     

Acid-sensing ion channel (ASIC) 1a/2a heteromers have a flexible 2:1/1:2 stoichiometry

Acid-sensing ion channels (ASICs) are widely expressed proton-gated Na⁺ channels playing a role in tissue acidosis and pain. A trimeric composition of ASICs has been suggested by crystallization. Upon coexpression of ASIC1a and ASIC2a in Xenopus oocytes, we observed the formation of heteromers and their coexistence with homomers by electrophysiology, but could not determine whether heteromeric complexes have a fixed subunit stoichiometry or whether certain stoichiometries are preferred over others. We therefore imaged ASICs labeled with green and red fluorescent proteins on a single-molecule level, counted bleaching steps from GFP and colocalized them with red tandem tetrameric mCherry for many individual complexes. Combinatorial analysis suggests a model of random mixing of ASIC1a and ASIC2a subunits to yield both 2:1 and 1:2 ASIC1a:ASIC2a heteromers together with ASIC1a and ASIC2a homomers.Acid-sensing ion channels (ASICs) are proton-gated Na⁺ channels, which are probably ubiquitously expressed in neurons, yet surprisingly little is known about their physiological function in the brain (1). It is believed that ASICs localize to the postsynapse and carry an excitatory postsynaptic current (2). ASICs in central neurons are mainly composed of homomeric ASIC1a and heteromeric ASIC1a/2a (3–6) and ASIC1a/2b (7). Genetic ablation of ASIC1a has indeed led to the conclusion that ASICs contribute to long-term potentiation and memory formation (2), but a more recent study challenged these findings (8). In contrast to our incomplete understanding of the physiological functions of ASICs, there is a comparatively large body of evidence that activation of ASIC1a-containing channels in pathophysiological states that are associated with sustained acidosis contributes to neuronal or axonal degeneration (9, 10). In addition, blockade of the ASIC1a/2a heteromer causes central analgesia (11). Together, these findings render ASIC1a-containing channels attractive drug target candidates.The crystal structure of chicken ASIC1 revealed an acidic pocket at subunit interfaces, which has been proposed to be the ligand-binding site of ASICs (12). In agreement, it has recently been shown that a gating modifier toxin of ASIC1, psalmotoxin 1, which behaves like an agonist (13), binds at the acidic pocket at subunit interfaces (14–16). Therefore, knowing the subunit composition of the ASIC1a/2a heteromer is of major interest, in particular for pharmacologically targeting this subtype.Fluorescence resonance energy transfer analysis suggested that the ASIC1a/2a heteromer contains at least two ASIC1a and two ASIC2a subunits (17). In agreement, several studies reported that the related epithelial Na⁺ channel (ENaC) is composed of four (18–21) or nine subunits (22–24). In strong contrast, however, the crystal structure of chicken ASIC1 revealed a number of three subunits (12, 25), suggesting that all ASICs and their relatives like ENaC are trimers. The trimeric structure of ASIC1a has in the meanwhile been confirmed by atomic force microscopy imaging (26). The stoichiometry of heteromeric ASICs, however, remains completely unknown.In this study, we first used electrophysiology to characterize mixtures of ASIC1a and ASIC2a at different expression ratios in Xenopus laevis oocytes to demonstrate that at least one heteromeric channel forms. Because we were unable to decide on the existence of a second heteromeric species by electrophysiology, we used a single-molecule photobleaching approach that resolves stoichiometries of membrane proteins with high accuracy. We tagged ASIC1a and ASIC2a with green and red fluorescent reporter proteins, which did not change the electrophysiological characteristics of homo- and heteromeric ASICs, suggesting that fusion of a fluorescent reporter has no impact on the molecular composition of ASICs. Colocalization of red and green reporter tags on a single-molecule level and counting of green bleaching steps confirms the trimeric nature of functional ASICs and shows that ASIC1a and ASIC2a randomly assemble into a complex with a flexible stoichiometry of either 1:2 or 2:1.