1.The effects of electroacupuncture on gastrointestinal motility and nitric oxide synthase positive nerves in myenteric plexus in acute exhaustive exercise rats
Chinese Journal of Physical Medicine and Rehabilitation 2008;30(4):248-251
Objective To study the effects of electroacupuncture(EA)on gastrointestinal motility and nitric oxide synthase(NOS)positive nerves in myenteric plexus in acute exhaustive exercise rats. Methods A total of 54 male Sprague-Dawley rats were randomly divided into an acute exhaustive exercise(AEE)group,an AEE plus EA group and a control(C)group.The gastric empty rate and intestinal motility rate were measured,and histological changes of NOS-positive nerves in ileum myenteric plexus were observed with enzymatic histochemical method and image analysis technique. Results In the rats of AEE group,the gastric empty rate and intestinal motility rate were delayed but the numbers of NOS-positive neurons and expression levels of NOS in the ileum myenteric plexus significantly increased comparing with those of the C group(P<0.01).Compared with AEE group,the gastric empty rate and intestinal motility rate were significantly increased but the number of NOS-positive neurons and expression level of NOS in the ileum myenteric plexus significantly decreased in AEE plus EA group(P<0.01). Conclusion The findings of this study suggest that increase of NOS-positive neurons and expression levels of NOS in the myenteric plexus is one of the mechanisms of slowing down of gastrointestinal movement after acute exhaustive exercise.Electroacupuncture at Zusanli acupoint may decrease the number of NOS-positive neurons and expression levels of NOS in the myenteric plexus,and improve gastrointestinal movement in acute exhaustive exercise rats.
2.Construction of antisense RNA expression plasmid for u-PAR and its transfection to highly invasive PC-3M cell subclones.
Guoning LIAO ; Qingfen LI ; Youmei FENG ; Yaozu DENG ; Zhuoya LI ; Feili GONG ; Ding MA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(4):369-372
To evaluate the specific inhibition of antisense u-PAR on the u-PAR expressions in highly invasive cell subclones and to determine its blocking function in the invasion by those cells, a cDNA fragment of u-PAR obtained by RT-PCR was inserted into a plasmid vector named pcDNA3 in antisense orientation. Then the antisense u-PAR recombinant was transfected into highly invasive cell subclones. The u-PAR expression in neo-resistant cells was examined by RT-PCR and immunohistochemical assay. Compared to the control cells, the content of mRNA and protein of u-PAR in transfected cells decreased sharply, and the rate of inhibition was 53% and 73%, respectively, indicating that an antisense u-PAR might have played a specific inhibitory role in its expression in the cells, which may provide a good cell model for making further investigation of the inhibitory effects of the antisense u-PAR on invasion in highly invasive cell subclones of human prostate carcinoma.
Cell Line, Tumor
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Cloning, Molecular
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Humans
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Male
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Neoplasm Invasiveness
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Plasmids
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Prostatic Neoplasms
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metabolism
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pathology
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RNA, Antisense
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biosynthesis
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genetics
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Receptors, Cell Surface
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biosynthesis
;
genetics
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Receptors, Urokinase Plasminogen Activator
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Reverse Transcriptase Polymerase Chain Reaction
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Transfection
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Urokinase-Type Plasminogen Activator
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antagonists & inhibitors
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genetics
;
metabolism
3.The inhibitory effects of an antisense u-PAR vector on invasion of highly invasive human prostate carcinoma PC-3M cell subclones.
Guoning LIAO ; Qingfen LI ; Youmei FENG ; Yaozu DENG ; Zhuoya LI ; Feili GONG ; Ding MA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(2):101-104
To observe the inhibitory effects of an antisense u-PAR vector on invasion of highly invasive PC-3M cell subclones, the effects of the antisense u-PAR on activity of MMP-9 in those highly invasive cell subclones were detected by a quantitative RT-PCR and zymography. The monolayer invasion assay and colony formation assay in soft agar were used. And tumorigenesis rate and invasions by the cell subclones with or without the antisense u-PAR were observed in nude mice. It was found that in vitro growth of highly invasive PC-3M cell subclones transfected with the antisense u-PAR was declined, and the ability of anchorage-independent growth of those cell subclones was found decreased sharply, with the inhibiting rate becoming 79% and 60%, respectively. Although the antisense u-PAR didn't change MMP-9 gene transcription, they could inhibit the activation of MMP-9 of highly invasive PC-3M cell subclones. Moreover, the tumorigenesis rate of the cell subclones with the antisense u-PAR decreased and the growth of a neoplasm also slowed down. The t tests showed the difference between experimental and control groups was statistically significant (P < 0.01). The antisense u-PAR vector could not only inhibit the invasion ability of highly invasive PC-3M cell subclones in vitro but also restrain the growth of those cell subclones in vivo.
Animals
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Antisense Elements (Genetics)
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genetics
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pharmacology
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Cell Division
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drug effects
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Cloning, Molecular
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Humans
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Male
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Matrix Metalloproteinase 9
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genetics
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metabolism
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Mice
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Mice, Inbred BALB C
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Mice, Nude
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Neoplasm Invasiveness
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Prostatic Neoplasms
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metabolism
;
pathology
;
RNA, Antisense
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Receptors, Cell Surface
;
genetics
;
metabolism
;
Receptors, Urokinase Plasminogen Activator
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Reverse Transcriptase Polymerase Chain Reaction
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Transfection
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Tumor Cells, Cultured