1.Effects of Fengliao-Changweikang in Diarrhea-predominant Irritable Bowel Syndrome Rats and Its Mechanism Involving Colonic Motility
Mengdi JIA ; Xiaofang LU ; Zhengfang WANG ; Luqing ZHAO ; Shengsheng ZHANG
Journal of Neurogastroenterology and Motility 2018;24(3):479-489
BACKGROUND/AIMS: This study was designed to investigate the effect of Fengliao-Changweikang (FLCWK) in diarrhea-predominant irritable bowel syndrome (IBS-D) rats and explore its underlying mechanisms. METHODS: IBS-D model rats were induced by neonatal maternal separation (NMS) combined with restraint stress (RS). In in vivo experiments, the model rats were randomly divided into 5 groups: NMS + RS, FLCWK (low dose, middle dose, and high dose), and pinaverium bromide. The normal control (no handling) rats were classified as the NH group. The therapeutic effect of FLCWK was evaluated by fecal characteristics, electromyographic response and abdominal withdrawal reflex scores. In in vitro experiments, the model rats were randomly divided into 2 groups: NMS + RS, FLCWK (middle dose), and no handling rats were used as the NH group. The differences in basic tension and ACh-induced tension of isolated colonic longitudinal smooth muscle strips (CLSMs) among the 3 groups were observed. In addition, different inhibitors (nifedipine, TMB-8, L-NAME, methylene blue, and 4-AP) were pretreated to explore the underlying mechanisms. RESULTS: In in vivo experiments, fecal characteristics, electromyographic response, and abdominal withdrawal reflex scores significantly improved in the FLCWK group, compared with the NMS + RS group. In in vitro experiments, the basic tension and ACh-induced tension of CLSMs in IBS-D rats were significantly inhibited by FLCWK. After pre-treatment with different inhibitors, the ACh-induced tension of CLSMs in each group showed no significant difference. CONCLUSIONS: FLCWK manifested curative effect in IBS-D rats by inhibiting colonic contraction. The underlying mechanisms may be related to regulatory pathway of nitric oxide/cGMP/Ca2+ and specific potassium channels.
Animals
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Colon
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Gastrointestinal Motility
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Herbal Medicine
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In Vitro Techniques
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Irritable Bowel Syndrome
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Methylene Blue
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Muscle, Smooth
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NG-Nitroarginine Methyl Ester
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Nitric Oxide
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Potassium Channels
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Rats
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Reflex
2.In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig
Huimin ZHAO ; Junyu NIE ; Xiangxing ZHU ; Yangqing LU ; Xingwei LIANG ; Huiyan XU ; Xiaogan YANG ; Yunkai ZHANG ; Kehuan LU ; Shengsheng LU
Journal of Veterinary Science 2018;19(5):592-599
In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.
Acrosin
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Activins
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Alkaline Phosphatase
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Bone Morphogenetic Protein 4
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Dolichos
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Follicle Stimulating Hormone
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Humans
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In Vitro Techniques
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Infant
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Infant, Newborn
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Spermatogenesis
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Stem Cells
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Swine
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Testis
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Testosterone
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Tretinoin
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Up-Regulation