Calcineurin-NFAT signaling is involved in phenylephrine-induced vascular smooth muscle cell proliferation

Aim： Catecholamine-induced vascular smooth muscle cell （VSMC） proliferation is one of the major events in the pathogenesis of atherosclerosis and vascular remodeling. The calcineurin-NFAT pathway plays a role in regulating growth and differentiation in various cell types. We investigated whether the calcineurin-NFAT pathway was involved in the regulation of phenylephrine-induced VSMC proliferation. Methods： Proliferation of VSMC was measured using an MTT assay and cell counts. Localization of NFATcl was detected by immunofluorescence staining. NFATcl-DNA binding was determined by EMSA and luciferase activity analyses. NFATcl and calcineurin levels were assayed by immunoprecipitation. Results： Phenylephrine （PE, an α1-adrenoceptor agonist） increased VSMC proliferation and cell number. Prazosin （an α1-adrenoceptor antagonist）, cyclosporin A （CsA, an inhibitor of calcineurin） and chelerythrine （an inhibitor of PKC） decreased PE-induced proliferation and cell number. Additional treatment of VSMC with CsA or chelerythrine further inhibited proliferation and cell number in the chelerythrine-pretreatment group and the CsA-pretreatment group. CsA and chelerythrine alone had no effect on either absorbance or cell number. CsA decreased PE-induced calcineurin levels and activity. NFATcl was translocated from the cytoplasm to the nucleus upon treatment with PE. This translocation was reversed by CsA. CsA decreased the PE-induced NFATcl level in the nucleus. PE increased NFAT＇s DNA binding activity and NFAT-dependent reporter gene expression. CsA blocked these effects. Conclusion： CsA partially suppresses PE-induced VSMC proliferation by inhibiting calcineurin activity and NFATcl nuclear translocation. The calcineurin-NFATcl pathway is involved in the hyperplastic growth of VSMC induced by phenylephrine.     

五味子乙素对慢性应激抑郁大鼠海马BDNF/TrkB/CREB通路的影响

目的 研究五味子乙素对慢性应激抑郁大鼠海马脑源性神经营养因子（BDNF）/酪氨酸激酶B（TrkB）/环磷腺苷效应元件结合蛋白（CREB）信号通路的影响。方法 40只SD大鼠随机选择10只作为对照组,其余大鼠采用慢性不可预知温和应激（chronic unpredictable mild stress,CUMS）结合孤养制备抑郁症模型,造模结束后随机分为3组：模型组、盐酸氟西汀（3 mg·kg^-1）组、五味子乙素（5 mg·kg^-1）组,每天ig给药1次,连续8周。分别于造模前、造模后及给药后进行旷场、悬尾、强迫游泳行为学实验;通过苏木素-伊红（HE）染色观察大鼠海马形态学改变;免疫组织化学染色（IHC）法观察大鼠海马BDNF蛋白表达;实时荧光定量PCR（qRT-PCR）法检测大鼠海马BDNF、TrkB、CREB mRNA相对表达量;Westernblotting检测大鼠海马BDNF、TrkB、CREB蛋白相对表达量。结果 与对照组比较,模型组大鼠旷场实验水平、垂直得分显著降低（P＜0.05）,悬尾不动时间和强迫游泳漂浮时间显著增加（P＜0.05）;HE染色结果显示海马神经元结构损伤,IHC结果显示海马BDNF表达明显降低;海马BDNF、TrkB、CREB mRNA及蛋白相对表达显著降低（P＜0.05）。与模型组比较,盐酸氟西汀及五味子乙素组大鼠水平、垂直得分显著增加（P＜0.05）,不动时间和漂浮时间显著减少（P＜0.05）;海马神经元结构明显复原,海马组织中BDNF染色明显增加;BDNF、TrkB、CREB mRNA和蛋白相对表达量显著增加（P＜0.05）。结论 五味子乙素可以改善慢性应激抑郁大鼠抑郁样行为、海马区神经元数量及形态,其机制可能与上调BDNF/TrkB/CREB信号通路有关。     

Effects of Total Flavones from Acanthopanax senticosus on L‐type Calcium Channels,Calcium Transient and Contractility in Rat Ventricular Myocytes

Acanthopanax senticosus (Rupr. et Maxim.) Harms (AS), a traditional herbal medicine, has been widely used to treat ischemic heart disease. However, the underlying cellular mechanisms of its benefits to cardiac function remain unclear. The present study examined the effects of total flavones from AS (TFAS) on L‐type Ca²⁺ channel currents (I_Ca‐L) using the whole cell patch‐clamp technique and on intracellular calcium ([Ca²⁺]_i) handling and cell contractility in rat ventricular myocytes with the aid of a video‐based edge‐detection system. Exposure to TFAS resulted in a concentration‐ and voltage‐dependent blockade of I_Ca‐L, with the half‐maximal inhibitory concentration (IC₅₀) of 283.12 µg/mL and the maximal inhibitory effect of 36.49 ± 1.95%. Moreover, TFAS not only increased the maximum current in the current–voltage relationship but also shifted the activation and inactivation curves of I_Ca‐L toward the hyperpolarizing direction. Meanwhile, TFAS significantly reduced amplitudes of myocyte shortening and [Ca²⁺]_i with an increase in the time to 10% of the peak (Tp) and a decrease in the time to 10% of the baseline (Tr). Thus, the cardioprotective effects of TFAS may be attributed mainly to the attenuation of [Ca²⁺]_i through the direct inhibition of I_Ca‐L in rat ventricular myocytes and consequent negative effect on myocardial contractility. Copyright © 2015 John Wiley & Sons, Ltd.     

Chemoprotective effect of plant phenolics against anthracycline-induced toxicity on rat cardiomyocytes. Part III. Apigenin, baicalelin, kaempherol, luteolin and quercetin

Flavonoids are found universally in plants and act as free radical scavenging and chelating agents with antiinflammatory, antiischemic, vasodilating and chemoprotective properties. In this study, the antilipoperoxidative and cytoprotective effects of apigenin, baicalein, kaempferol, luteolin and quercetin against doxorubicin-induced oxidative stress were investigated in isolated rat heart cardiac myocytes, mitochondria and microsomes. After preincubation of cardiomyocytes with the test compounds for 1 h the cardiomyocytes were treated with the toxic agent, doxorubicin (100 micro M for 8 h). Cardiomyocyte protection was assessed by extracellular LDH and cellular ADP and ATP production. Cytoprotection was concentration dependent for baicalein > luteolin congruent with apigenin > quercetin > kaempferol. All test compounds had signi fi cantly better protective effects than dexrazoxan, an agent currently used for adjuvant therapy during anthracycline antibiotic therapy. In microsomes/mitochondria the IC(50) values of lipid peroxidation inhibition for quercetin, baicalein, kaempferol, luteolin, and apigenin were 3.1 +/- 0.2/8.2 +/- 0.6, 3.3 +/- 0.3/9.6 +/- 0.5, 3.9 +/- 0.3/10.1 +/- 0.8, 22.9 +/- 1.7/18.2 +/- 0.7, and 338.8 +/- 23.1/73.1 +/- 6.4 mM, respectively. The antilipoperoxidative activity of apigenin differed from its cytoprotective effects, but correlated with the free radical scavenging of 2,2-diphenyl-1-picrylhydrazyl radical and half peak oxidation potential (E(p/2)). Apigenin was the least effective of the flavonoids studied in all models except the cardiomyocyte model where its cardiomyocyte cytoprotective effect was comparable to other compounds.     

7,7'-Dimethoxyagastisflavone-induced apoptotic or autophagic cell death in different cancer cells

7,7'-Dimethoxyagastisflavone (DMGF), a biflavonoid isolated from the needles of Taxus × media cv. Hicksii, was evaluated for its antiproliferative and antineoplastic effects in three human cancer cell lines. Interestingly, DMGF caused cell death via different pathways in different cancer cells. DMGF induced apoptosis, activated caspase-3 activity and changed the mitochondrial membrane potential in HT-29 human colon cancer cells. However, the apoptotic pathway is not the major pathway involved in DMGF-induced cell death in A549 human lung cancer cells and HepG2 human hepatoma cells. Treatment with 3-MA, an inhibitor of autophagy, significantly decreased DMGF-induced cell death in HepG2 and A549 cells, but did not affect DMGF-induced cell death in HT-29 cells. Following DMGF treatment, the HepG2 cells increased expression of LC3B-II, a marker used to monitor autophagy in cells. Thus, DMGF induced apoptotic cell death in HT-29 cells, triggered both apoptotic and autophagic death in A549 cells and induced autophagic cell death in HepG2 cells.     

肿瘤突变负荷位点检测准确性评价

目的 使用肿瘤突变负荷国家参考品中高置信SNP/Indel位点标准集的参考品,评价肿瘤突变负荷检测试剂盒的位点检测准确性.方法 采用TMB检测试剂盒(可逆末端终止测序法)检测肿瘤突变负荷检测国家参考品中高置信SNP/Indel位点标准集的参考品.首先将DNA片段化处理,进行末端修复、接头连接和文库扩增等步骤后构建文库;...     

Investigation of the larvicidal activity of Pothomorphe peltata and isolation of the active constituent

The insecticidal activity of the leaves of Pothomorphe peltata (Piperaceae) was evaluated on Aedes aegypti larvae. The active methanol extract was subjected to a bioactivity-directed fractionation, monitored by the larvicidal assay, and led to the isolation of a catechol derivative, 4-nerolidylcatechol. This compound was quite potent against Aedes aegypti larvae (LC(50) = 9.1 microg/mL).     

陈红风治疗乳腺癌中的扶正观

介绍陈红风教授治疗乳腺癌的临床经验。认为乳腺癌的发病与气血亏虚,肝、脾、肾及冲任二脉功能失调有关;治疗时重视通过调摄冲任以扶正、调理脏腑以祛邪。并附病例1则。     

Selective Antibacterial Activity of Patchouli Alcohol Against Helicobacter pylori Based on Inhibition of Urease

The aim of this study is to evaluate the antibacterial activity and urease inhibitory effects of patchouli alcohol (PA), the bioactive ingredient isolated from Pogostemonis Herba, which has been widely used for the treatment of gastrointestinal disorders. The activities of PA against selected bacteria and fungi were determined by agar dilution method. It was demonstrated that PA exhibited selective antibacterial activity against Helicobacter pylori, without influencing the major normal gastrointestinal bacteria. Noticeably, the antibacterial activity of PA was superior to that of amoxicillin, with minimal inhibition concentration value of 78 µg/mL. On the other hand, PA inhibited ureases from H. pylori and jack bean in concentration‐dependent fashion with IC₅₀ values of 2.67 ± 0.79 mM and 2.99 ± 0.41 mM, respectively. Lineweaver‐Burk plots indicated that the type of inhibition was non‐competitive against H. pylori urease whereas uncompetitive against jack bean urease. Reactivation of PA‐inactivated urease assay showed DL‐dithiothreitol, the thiol reagent, synergistically inactivated urease with PA instead of enzymatic activity recovery. In conclusion, the selective H. pylori antibacterial activity along with urease inhibitory potential of PA could make it a possible drug candidate for the treatment of H. pylori infection. Copyright © 2014 John Wiley & Sons, Ltd.     

The successful use of alemtuzumab for treatment of steroid‐refractory acute graft‐versus‐host disease in pediatric patients

SR‐aGVHD remains a significant cause of morbidity and mortality in allogeneic HCT recipients. Alemtuzumab has been used with success in adult patients but has not been studied in the pediatric setting. To estimate the effectiveness of alemtuzumab for the treatment of SR‐aGVHD in pediatric patients, we retrospectively reviewed the charts of 19 patients (median age 4 yr, range 0.5–28 years) with grades II (n = 3), III (n = 10), or IV (n = 6) SR‐aGVHD who received alemtuzumab treatment. Patients received a median dose of 0.9 mg/kg alemtuzumab (range 0.3–2 mg/kg) divided over 2–6 days. Eighty‐nine percent of patients received additional courses. A complete response, defined as GVHD of grade 0 at four wk following the first alemtuzumab course, was observed in nine patients (47%). A partial response, defined as an improvement in grade after four wk, was observed in five patients (26%). There was no response in five patients (26%). The overall response rate at four wk was 73%. Infectious complications included bacteremia (47%), presumed or documented fungal infections (21%), adenovirus viremia (52%), EBV viremia (36%), and CMV viremia (36%). We conclude that alemtuzumab is effective for SR‐aGVHD in pediatric patients with a tolerable spectrum of complications.