Repression of miR-142–3p alleviates psoriasis-like inflammation by repressing proliferation and promoting apoptosis of keratinocytes via targeting Sema3A

Psoriasis is a multifactorial, recurring, and chronic inflammatory skin disease characterized by hyperproliferation of keratinocytes. Evidence is rapidly accumulating for the role of microRNAs in psoriasis. The object of the study was to explore the functions and precise mechanism of miR-142–3p in human keratinocyte HaCaT cells in the presence of M5. Here, the results showed that miR-142–3p expression was heightened in HaCaT cells induced by M5. In addition, inhibition of miR-142–3p dramatically restricted cell proliferation and enhanced apoptosis in HaCaT cells exposed to M5, as exemplified by a decrease in the antiapoptotic Bcl-2 protein, concomitant with an increase in the proapoptotic proteins Bax. Moreover, depleting miR-142–3p effectively ameliorated M5-induced inflammation response, as reflected by the attenuation of multiple inflammatory factors. Importantly, Sema3A was identified as an authentic target of miR-142–3p, and indeed regulated by miR-142–3p. Mechanistically, silencing of Sema3A effectively abolished the anti-proliferative, apoptosis-promoting, and anti-inflammatory effects of miR-142–3p inhibition in keratinocytes. Taken together, these data elucidated that repression of miR-142–3p protect HaCaT cells against M5-induced hyper-proliferation and inflammatory injury by suppressing its target Sema3A, implying that the miR-142–3p/Sema3A axis may be a new target for preventing keratinocyte injury process. These findings provide a new and better understanding of the mediating role of miR-142–3p in psoriasis.     

基于PI3K/AKT信号通路的谷红注射液抗H9c2心肌细胞缺氧/复氧损伤的机制研究

目的基于PI3K/AKT信号通路,探讨谷红注射液(guhong injection,GHI)对H9c2心肌细胞缺氧/复氧(H/R)损伤的保护作用。方法培养H9c2心肌细胞,CCK-8比色法筛选GHI实验剂量。将细胞随机分为8组:正常组、模型组、GHI低、中、高剂量组(30、60和90μL·mL^-1)、阳性药组(维拉帕米注射液7.5μL·mL^-1)、GHI+LY294002(PI3K抑制剂)组和LY294002组。除正常组外,其他组均建立缺氧4 h复氧16 h的H/R损伤模型。ELISA检测各组细胞内肌酸激酶同工酶(CKMB)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量,Western blot法检测各组细胞内p-PI3K、PI3K、p-AKT、AKT及GSK-3β蛋白表达水平。结果与模型组相比,各给药组LDH、CK-MB均降低(P<0.05),MDA降低(P<0.01)、SOD升高(P<0.01)。p-PI3K/PI3K、p-AKT/AKT及GSK-3β蛋白表达水平均升高(P<0.01)。给予LY294002后,与模型组相比,LDH、CK-MB、MDA及SOD无显著性差异,p-PI3K/PI3K、p-AKT/AKT及GSK-3β蛋白表达水平均降低(P<0.01)。结论GHI可以明显减轻H/R对H9c2心肌细胞造成的损伤,表现出抗氧化应激与抗凋亡的作用,其作用机制可能与PI3K/AKT信号通路有关。     

HSP90β干扰和过表达慢病毒载体的构建及其在骨肉瘤细胞Saos-2中的表达

目的:获得热休克蛋白90β(HSP90β)基因干扰和过表达慢病毒表达系统，并检测其在人骨肉瘤细胞株Saos-2中的表达水平。方法:设计合成shRNA，以慢病毒表达质粒构建HSP90β干扰和过表达载体，酶切电泳、测序技术鉴定载体构建是否成功。重组病毒转染H1299细胞，以嘌呤霉素筛选稳定转染的Saos-2细胞，通过荧光显微镜观察计数获得转染效率。将感染好的细胞分为干扰NC组（NEG-shRNA）、干扰组（shRNA-HSP90β）、过表达NC对照组（NEG-pEZ）及过表达组（pEZ-HSP90β）。通过qRT-PCR 与Western blotting 分别从mRNA 和蛋白表达水平验证目的基因的干扰和过表达水平。结果:插入慢病毒表达载体的基因片段与目的基因的碱基序列完全一致。病毒包装成功后，嘌呤霉素最小致死浓度1 μg/ml，感染复数200，感染人Saos-2的感染效率达80%，其中shRNA-HSP90β组干扰效率为86.35%，pEZ-HSP90β组的mRNA相对表达量增加2.8倍。进一步研究发现，shRNA-HSP90β组较NEG-shRNA组中HSP90β蛋白表达明显降低，pEZ-HSP90β组较NEG-pEZ组HSP90β蛋白表达增加。结论:HSP90β基因干扰和过表达慢病毒载体构建成功，并能够在Saos-2中稳定表达。     

KIF20A对胶质瘤细胞增殖、迁移和侵袭、凋亡及细胞周期分布的影响

目的:研究KIF20A对胶质瘤U87细胞系增殖、侵袭、迁移、凋亡及细胞周期分布的影响。方法:利用 RNA干扰技术下调胶质瘤细胞系 U87中 KIF20A 的表达。RT-qPCR 和 Western blot 分别检测 KIF20A 在mRNA 和蛋白水平上的表达。CCK-8 实验检测 U87细胞增殖能力的变化，Transwell 实验检测U87细胞迁移及侵袭能力的变化，流式细胞术检测 U87细胞凋亡及细胞周期的变化。结果:RT-qPCR和Western blot 结果显示 siRNA-KIF20A显著下调 KIF20A 的表达，CCK-8和Transwell 实验显示下调 KIF20A 的表达显著抑制了U87细胞的增殖、迁移及侵袭能力，流式细胞术结果显示下调 KIF20A 的表达显著促进了U87细胞的凋亡，诱导细胞周期阻滞在G0/G1期。结论:下调KIF20A的表达抑制胶质母细胞瘤细胞的增殖、迁移及侵袭，促进胶质母细胞瘤细胞的凋亡并诱导细胞周期G0/G1期阻滞。     

丝穗金粟兰化学成分研究

目的对丝穗金粟兰Chloranthus fortunei的化学成分进行研究。方法利用多种色谱分离方法和波谱学鉴定方法对丝穗金粟兰中的化学成分进行了分离鉴定,并借助MTT法对其中得到的部分化合物进行了抗肿瘤活性的初步筛选。结果从丝穗金粟兰95%乙醇提取物中分离得到16个化合物,分别鉴定为迷迭香酸(1)、2′-羟基-4,3′,4′,6′-四甲氧基查耳酮(2)、卡瓦胡椒素A(3)、cycloshizukaol A(4)、白术内酯III(5)、4β-hydroxy-8,12-epoxyeudesma-7,11-diene-1,6-dione(6)、(8α)-6,8-dihydroxycadina-7(11),10(15)-dien-12-oic acidγ-lactone(7)、curcolonol(8)、11-hydroxyldrim-8,12-en-14-oic acid(9)、木栓酮(10)、异香草酸(11)、6β-hydroxystigmast-4-en-3-one(12)、3,4-二羟基苯甲酸(13)、莽草酸(14)、东莨菪苷(15)以及N-acetyltyramine 1-O-β-D-glucoside(16)。化合物4和5表现出微弱的细胞毒作用,半数抑制浓度(IC50)值在46～85μmol/L。结论化合物2、10、11、13～15为首次从金粟兰属植物中获得,化合物1～3、6～16为首次从丝穗金粟兰中分离得到。丝穗金粟兰中部分倍半萜显示出弱的抗肿瘤活性。     

益智仁化学成分研究

目的研究益智Alpinia oxyphylla仁的化学成分。方法采用硅胶、MCI柱色谱、Sephadex LH-20、重结晶、半制备液相进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。结果从益智仁95%乙醇提取物的醋酸乙酯部位分离并鉴定16个化合物,分别鉴定为邻苯二甲酸-双(2′-乙基庚基)酯(1)、(E)-1-(4′-hydroxy-3′-methoxyphenyl)-7-(4″-hydroxyphenyl)-hept-4-en-3-one(2)、5-hydroxy-7-(4″-hydroxy-3″-methoxyphenyl)-1-phenyl-3-heptanone(3)、1β,4β,7β-trihydroxyeudesmane(4)、bullatantriol(5)、1,5-epoxy-3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl) heptanes(6)、1-tetratriacontanol(7)、dihydrogingerenone B(8)、杨芽黄素(9)、白杨素(10)、oxyphyllenone B(11)、(1R,4R,10R)-1β,4α-dihydroxy-11,12,13-trinor-5,6-eudesmen-7-one(12)、胡萝卜苷(13)、1-(4′-羟基苯基)-7-(3″-甲氧基-4″-羟基苯基)-4-烯-3-庚酮(14)、益智酮甲(15)、5-dehydroxy-hexahydro-demethoxycurcumin B(16)。结论其中化合物1、2、4～8为首次从该属植物中分离得到,化合物3为首次从该植物中分离得到。     

Combinatorial Inhibition of Myostatin and Activin A Improves Femoral Bone Properties in the G610C Mouse Model of Osteogenesis Imperfecta

Osteogenesis imperfecta (OI) is a collagen-related bone disorder characterized by fragile osteopenic bone and muscle weakness. We have previously shown that the soluble activin receptor type IIB decoy (sActRIIB) molecule increases muscle mass and improves bone strength in the mild to moderate G610C mouse model of OI. The sActRIIB molecule binds multiple transforming growth factor-β (TGF-β) ligands, including myostatin and activin A. Here, we investigate the musculoskeletal effects of inhibiting activin A alone, myostatin alone, or both myostatin and activin A in wild-type (Wt) and heterozygous G610C (+/G610C) mice using specific monoclonal antibodies. Male and female Wt and +/G610C mice were treated twice weekly with intraperitoneal injections of monoclonal control antibody (Ctrl-Ab, Regn1945), anti-activin A antibody (ActA-Ab, Regn2476), anti-myostatin antibody (Mstn-Ab, Regn647), or both ActA-Ab and Mstn-Ab (Combo, Regn2476, and Regn647) from 5 to 16 weeks of age. Prior to euthanasia, whole body composition, metabolism and muscle force generation assessments were performed. Post euthanasia, hindlimb muscles were evaluated for mass, and femurs were evaluated for changes in microarchitecture and biomechanical strength using micro–computed tomography (μCT) and three-point bend analyses. ActA-Ab treatment minimally impacted the +/G610C musculoskeleton, and was detrimental to bone strength in male +/G610C mice. Mstn-Ab treatment, as previously reported, resulted in substantial increases in hindlimb muscle weights and overall body weights in Wt and male +/G610C mice, but had minimal skeletal impact in +/G610C mice. Conversely, the Combo treatment outperformed ActA-Ab alone or Mstn-Ab alone, consistently increasing hindlimb muscle and body weights regardless of sex or genotype and improving bone microarchitecture and strength in both male and female +/G610C and Wt mice. Combinatorial inhibition of activin A and myostatin more potently increased muscle mass and bone microarchitecture and strength than either antibody alone, recapturing most of the observed benefits of sActRIIB treatment in +/G610C mice. © 2022 American Society for Bone and Mineral Research (ASBMR).     

不同产地当归藤HPLC指纹图谱

目的建立不同产地当归藤HPLC指纹图谱。方法当归藤85%甲醇提取物的分析采用Phenomenex C18色谱柱(4.6 mm×250 mm,5μm);流动相乙腈⁃0.1%磷酸水,梯度洗脱;体积流量0.8 mL/min;柱温30℃;检测波长225 nm。对结果进行聚类分析和主成分分析。结果12批样品指纹图谱中有12个共有峰,相似度均大于0.940,并指认了没食子酸,儿茶素;聚类分析将12批样品分为2类。结论该方法准确稳定,重复性好,可用于当归藤药材的质量控制。     

养阴利咽饮与玉液散吹喉对阴虚肺燥型慢性咽炎患者血清因子的影响

目的:探讨养阴利咽饮与玉液散吹喉对阴虚肺燥型慢性咽炎患者血清炎性因子的影响。方法:将2015年8月至2017年3月山东省淄博市第四人民医院收治的慢性咽炎患者118例作为研究对象,按治疗方法不同分为对照组和观察组,每组59例。对照组用常规西药治疗,观察组用养阴利咽饮与玉液散吹喉治疗,均连续治疗30 d。比较2组临床疗效,观察并比较治疗前后2组患者主要临床症状改善情况,唾液中分泌型免疫球蛋白A(SIgA)、血清炎性因子水平及生命质量变化。结果:治疗后观察组总有效率为91.53%,高于对照组的76.27%(P<0.05)。治疗后2组咽痛、干咳、咽黏膜充血水肿临床症状积分低于治疗前,且观察组低于对照组(P<0.01)。与治疗前比较,治疗后2组患者唾液中SIgA水平均明显升高,血清肿瘤坏死因子-α(TNF-α)及C-反应蛋白(CRP)水平均明显降低,且2组比较差异均有统计学意义(均P<0.01)。治疗后2组患者SF-36各项评分均明显高于治疗前,且观察组高于对照组(P<0.05或P<0.01)。结论:养阴利咽饮与玉液散吹喉联用可降低阴虚肺燥型慢性咽炎患者血清TNF-α、CRP水平,升高唾液中SIgA含量,改善患者临床症状,提高患者生命质量。