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Zipeng Gong,Ying Chen,Ruijie Zhang,Qing Yang,Yajie Wang,Yan Guo,Bingbing Zhou,Xiaogang Weng,Xuchen Liu,Yujie Li,Xiaoxin Zhu,Yu Dong 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.10
Berberine is one of active alkaloids from Rhizomacoptidis in traditional Chinese medicine. The pharmacokineticsof berberine in rat plasma were compared betweennormal and chronic visceral hypersensitivity irritable bowelsyndrome rats (CVH-IBS) established by mechanical colonirritation using angioplasty balloons for 2 weeks after oraladministration of berberine hydrochloride (25 mg/kg) withthe equivalent dose of 22 mg/kg for berberine according tobody weight. Immunohistochemical analysis of c-fos andmyosin light chain kinase (MLCK) and immunofluorescenceanalysis of MLCK in rat colon were conducted. Quantificationof berberine in rat plasma was achieved by using a sensitiveand rapidUPLC-MS/MSmethod. Plasma samples werecollected at 15 different points in time and the pharmacokineticparameters were analyzed by WinNonlin software. Thegreat different pharmacokinetic behavior of berberine wasobserved between normal and CVH-IBS model rats. Compared with normal group, T1/2 and AUC(0–t) of berberinein the model group were significantly increased, respectively(573.21 ± 127.53 vs 948.22 ± 388.57 min; 8,657.19 ±1,562.54 vs 11,415.12 ± 1,670.72 min.ng/ml). Cl/F of berberinein the model group significantly decreased, respectively(13.89 ± 1.69 vs 9.19 ± 2.91 L/h/kg). Additionally,the expressions of c-fos and MLCK in model group werehigher than those in normal group. The pharmacokinetic behaviorof berberine was significantly altered in CVH-IBSpathological conditions, which indicated the dosage modificationof berberine hydrochloride in CVH-IBS were necessary. Especially, improved exposure to berberine in ratplasma inCVH-IBSmodel rats was attributed to increased theexpression of MLCK.
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Jiawei Lv,Shuang Wu,Renyue Wei,Yan Li,Junxue Jin,Yanshuang Mu,Yu Zhang,Qingran Kong,Xiaogang Weng,Zhonghua Liu 대한수의학회 2019 Journal of Veterinary Science Vol.20 No.3
The clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a versatile genome editing tool with high efficiency. A guide sequence of 20 nucleotides (nt) is commonly used in application of CRISPR/Cas9; however, the relationship between the length of the guide sequence and the efficiency of CRISPR/ Cas9 in porcine cells is still not clear. To illustrate this issue, guide RNAs of different lengths targeting the EGFP gene were designed. Specifically, guide RNAs of 17 nt or longer were sufficient to direct the Cas9 protein to cleave target DNA sequences, while 15 nt or shorter guide RNAs had loss-of-function. Full-length guide RNAs complemented with mismatches also showed loss-of-function. When the shortened guide RNA and target DNA heteroduplex (gRNA:DNA heteroduplex) was blocked by mismatch, the CRISPR/Cas9 would be interfered with. These results suggested the length of the gRNA:DNA heteroduplex was a key factor for maintaining high efficiency of the CRISPR/Cas9 system rather than weak bonding between shortened guide RNA and Cas9 in porcine cells.
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