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补肾方含药血清干预大鼠椎间盘软骨细胞miRNA的差异表达 被引量:8
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作者 潘欣 梁兴伦 +9 位作者 曾思良 嵇承栋 周文锐 李琛 朱敏洁 许畅 付强强 万悦竹 马瑜 于德华 《上海中医药杂志》 2016年第1期65-69,共5页
目的筛选补肾方含药血清干预大鼠椎间盘软骨细胞后差异表达的miRNA,为补肾方治疗颈椎病提供可供选择的目标miRNA,以探讨其通过调控靶基因表达发挥治疗效应的机制。方法原代培养的大鼠椎间盘软骨细胞经Ⅱ型胶原酶免疫荧光染色和甲苯胺蓝... 目的筛选补肾方含药血清干预大鼠椎间盘软骨细胞后差异表达的miRNA,为补肾方治疗颈椎病提供可供选择的目标miRNA,以探讨其通过调控靶基因表达发挥治疗效应的机制。方法原代培养的大鼠椎间盘软骨细胞经Ⅱ型胶原酶免疫荧光染色和甲苯胺蓝染色鉴定后,分别用补肾方含药血清、空白血清干预椎间盘软骨细胞,在给定时间点收获细胞,抽提纯化总RNA,行miRNA芯片检测,通过芯片差异性分析,挑选差异表达4倍以上的miRNA行定量聚合酶链式反应(PCR)验证。结果二次酶消化法可简便高效培养大量原代椎间盘软骨细胞,所培养细胞具有典型的椎间盘软骨细胞形态和功能,Ⅱ型胶原酶免疫荧光染色和甲苯胺蓝染色均为阳性。补肾方含药血清干预组芯片数据分别与相应时间点的空白血清组芯片数据行组间比较后再行组内比较,与含药血清干预椎间盘软骨细胞1 d相比,干预3 d的共有121个2倍以上表达差异的miRNA,其中上调的53个,下调的68个。差异表达4倍以上的miRNA定量PCR结果与芯片数据一致。结论补肾方含药血清干预椎间盘软骨细胞获得了一套较为特异的miRNA,初步预测上调的miRNA可能通过靶向Opn、Col、Bcl212、Vdr、IGF-I、Mmp-13等基因在椎间盘营养代谢相关信号通路中发挥作用。 展开更多
关键词 补肾方 软骨细胞 大鼠miRNA芯片 差异表达
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大鼠成骨细胞微小RNAs的筛选研究 被引量:2
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作者 潘欣 曾思良 +6 位作者 梁兴伦 嵇承栋 周文博 周文锐 李琛 朱敏洁 沈琪 《转化医学杂志》 2017年第2期68-73,共6页
目的应用微小RNAs(microRNAs,miRNAs)芯片技术筛选补肾方含药血清干预大鼠成骨细胞后差异表达的miRNAs,与前期获得的软骨细胞差异miRNAs比较,筛选出共有差异miRNAs,以探讨补肾方通过共有miRNAs调控靶基因表达发挥治疗效应的机制。方法... 目的应用微小RNAs(microRNAs,miRNAs)芯片技术筛选补肾方含药血清干预大鼠成骨细胞后差异表达的miRNAs,与前期获得的软骨细胞差异miRNAs比较,筛选出共有差异miRNAs,以探讨补肾方通过共有miRNAs调控靶基因表达发挥治疗效应的机制。方法原代培养的大鼠成骨细胞经碱性磷酸酶染色、Ⅰ型胶原免疫荧光染色和矿化结节染色鉴定后,用补肾方含药血清干预成骨细胞,在给定时间点收获细胞,抽提纯化总RNAs行miRNAs芯片检测,并挑选差异表达4倍以上的miRNAs行实时荧光定量逆转录-聚合酶链反应验证。结果所培养的原代细胞具有典型的成骨细胞形态和功能,碱性磷酸酶染色、Ⅰ型胶原免疫荧光染色和矿化结节染色均为阳性。与补肾方含药血清干预成骨细胞1 d相比,干预6 d的共有229个2倍以上表达差异的miRNAs,其中上调的52个、下调的177个。差异表达4倍以上的miRNAs实时荧光定量逆转录-聚合酶链反应结果与芯片数据一致。与前期软骨细胞差异miRNAs芯片数据比较,miRNAs均上调2倍以上的有8个、下调4倍以上的2个。结论补肾方含药血清干预成骨细胞获得了一套较为特异的miRNAs,初步预测上调的miRNAs可能通过靶向Runx2、Wnt、Notch、Axin2等基因在骨发育、骨形态发生、骨吸收以及软骨内成骨形成等相关的信号通路中发挥作用。 展开更多
关键词 补肾方 成骨细胞 微小RNAs芯片 差异表达 大鼠
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Manual acupuncture for patients with major depressive disorder and comorbid insomnia:Evidence from polysomnography and serum biomarkers 被引量:3
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作者 Fei-yi ZHAO Yan XU +6 位作者 Li-ping YUE Ying-xia ZHAO Ying WANG Hua-ling SONG Hong XU Si-han CHEN Qiang-qiang FU 《World Journal of Acupuncture-Moxibustion》 CSCD 2020年第1期5-12,共8页
Objective:This study is conducted to determine effects of manual acupuncture(MA)in patients with a diagnosis of major depressive disorder(MDD)and comorbid insomnia.Methods:A total of 67 participants who met the inclus... Objective:This study is conducted to determine effects of manual acupuncture(MA)in patients with a diagnosis of major depressive disorder(MDD)and comorbid insomnia.Methods:A total of 67 participants who met the inclusion criteria were randomly enrolled in a two-arm randomized,placebo controlled,patients-blind trial and allocated to a real-MA group(patients=34)and a sham-MA group(patients=33).Patients in the real-MA group were treated on‘Five-shen acupoints’,including Sishencong(四神聪EX-HN 1),Shenting(神庭GV 24),Shendao(神道GV 11),bilateral Benshen(本神GB 13),and bilateral Shenmen(神门HT 7).Though being punctured on the same acupoints,patients in the sham-MA group were treated by a placebo acupuncture approach(Streitberger Placeboneedle).Each group received corresponding interventions every other day,three times a week for total eight weeks.Both polysomnography(PSG)and testing of serum biological markers such as neuropeptide Y(NPY)and substance P(SP)were performed at pre-and post-treatment.Additionally,the global scores of Pittsburgh sleep quality index(PSQI)and the global scores of 17-items Hamilton Depression Rating Scale(HAMD17)were used for assessing the subjective sleep and emotion experience of patients,respectively.Meanwhile,adverse effects were monitored and recorded.Results:After eight-week treatment,the global scores of PSQI and global scores of HAMD17 declined significantly(both P<0.05)in the real-MA group but not in the sham-MA group(both P>0.05).According to the parameters of PSG,striking decline were observed in sleep latency(SL)and wake after sleep onset(WASO)and striking climb were observed in total sleep time(TST)and sleep efficiency(SE)in the real-MA group after treatment(P<0.05,respectively)but not in the sham-MA group(P>0.05,respectively).Additionally,there were no significant differences in awakening times(ATs)and rapid eyes movement sleep latency(REM-SL)in both two groups after treatment(both P>0.05).Meanwhile,the expression of NPY increased significantly and the expression of SP decreased significantly in the real-MA group after interventions(both P<0.05)while those indicators only slightly fluctuated in the sham-MA group(P>0.05).No serious adverse event was reported in either real-or sham-MA group.Conclusion:(1)MA may be a potential alternative therapy for improving MDD and comorbid insomnia(particularly in extending total sleep time and shortening wake-up duration and sleep latency)via upregulating the expression of NPY and downregulating the expression of SP;more importantly,this efficacy of acupuncture can not be replaced by sham-acupuncture acting on the same acupoints with the same treatment frequency.(2)There is insufficient evidence to prove that MA can effectively reduce the number of arousals. 展开更多
关键词 Major DEPRESSIVE DISORDER INSOMNIA Manual acupuncture POLYSOMNOGRAPHY NEUROPEPTIDE Y Substance P
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SOOJI CHIM (Koryo hand acupuncture) for perimenopausal insomnia:Evidence from actigraphy and serum hormone testing 被引量:4
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作者 Fei-Yi Zhao Jin-Hyeok Kim +6 位作者 Li-Ping Yue Hai-Xia Yan Hua-Ling Song Yan Xu Hong Xu Yu-Fang Hong Qiang-Qiang Fu 《World Journal of Acupuncture-Moxibustion》 CSCD 2019年第4期274-278,共5页
Objective:This study is conducted to determine the effects of SOOJI CHIM(Koryo hand acupuncture,KHA)therapy on sleep structures and perimenopause symptoms among insomniacs.Methods:Totally 49 participants who met the i... Objective:This study is conducted to determine the effects of SOOJI CHIM(Koryo hand acupuncture,KHA)therapy on sleep structures and perimenopause symptoms among insomniacs.Methods:Totally 49 participants who met the inclusion criteria were enrolled in a two-arm randomized,placebo controlled,patients-blind trial,with 24 cases allocated to a real-KHA group and 25 cases allocated to a sham-KHA group.Patients in each group received corresponding treatment every other day,three times a week for total eight weeks.Both actigraphy and testing of serum hormones level reflected by biological markers such as follicle-stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E2)were performed at pre-and post-treatment.Additionally,the scores of Pittsburgh sleep quality index(PSQI)was used for assessing the subjective sleep experience of insomniacs.Meanwhile,adverse effects were monitored and recorded.Results:After eight-week treatment,the global scores of PSQI declined significantly(P<0.05)in the realKHA group but not in the sham-KHA group(P>0.05).According to the outcomes of actigraphy,a striking increase was observed in both sleep efficiency(SE)and total sleep time(TST)in the real-KHA group(both P<0.05)but not in the sham-KHA group(P>0.05).However,no significant changes of sleep awakenings(SA)in both groups was found after treatment(both P>0.05).Meanwhile,FSH and LH in the real-KHA group decreased markedly after intervention(both P<0.05)while those indicators only slightly changed in the sham-KHA group(P>0.05).The fluctuation of E2 levels were not significant in both groups(both P>0.05).No serious adverse event was reported in either real-or sham-KHA group.Conclusion:KHA may be a potential alternative therapy for improving perimenopausal insomnia via down regulating the levels of FSH and LH. 展开更多
关键词 Perimenopausal insomnia SOOJI CHIM(Koryo hand acupuncture KHA) ACTIGRAPHY Serum hormone testing
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