1.Effect of Changji'an Formula (肠激安方) on the miR-29b-3p/TRAF3/NF-κB/MLCK Axis in Colonic Tissues in Diarrhea-predominant Irritable Bowel Syndrome Model Rat with Liver Depression and Spleen Deficiency Syndrome
Yongfu WANG ; Wei KE ; Xiangyu XIE ; Hongmei TANG ; Liuze SI ; Yuna CHAI
Journal of Traditional Chinese Medicine 2026;67(4):439-446
ObjectiveTo explore the potential mechanism of Changji'an Formula (肠激安方) on intestinal permeability for rats with diarrhea-predominant irritable bowel syndrome (IBS-D) of liver depression and spleen deficiency syndrome by the microRNA-29b-3p (miR-29b-3p)/tumor necrosis factor receptor-associated factor 3 (TRAF3)/nuclear factor-κB (NF-κB)/myosin light chain kinase (MLCK) axis. MethodsTwenty-four 1-day-old male Sprague-Dawley (SD) suckling rats were selected, and the IBS-D rat model of liver depression and spleen deficiency syndrome was established via a three-factor method,i.e. maternal separation plus acetic acid stimulation and restraint stress, for 6 consecutive weeks. After successful modeling, the rats were randomly divided into a model group, pinaverium bromide group, low-dose and high-dose Changji'an Formula groups, with 6 rats in each group. Another 6 age-matched non-modeled SD rats were included as the control group. The low-dose and high-dose Changji'an Formula groups were given intragastric administration of Changji'an Formula solution at doses of 16.74 g/(kg·d) and 33.48 g/(kg·d), respectively; the pinaverium bromide group received intragastric administration of pinaverium bromide tablets at 0.018 g/(kg·d); and the control group was given distilled water at 10 ml/(kg·d) via intragastric gavage. The intervention was conducted once daily for 14 consecutive days. After the gavage treatment, the fecal water content of rats in each group was measured. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum levels of intestinal permeability indicators, including D-lactic acid (D-LA), diamine oxidase (DAO), and lipopolysaccharide (LPS). Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to determine the mRNA expression levels of miR-29b-3p, TRAF3, tumor necrosis factor-α (TNF-α), p65, p50, and MLCK in colonic tissues. Western Blot analysis was employed to detect the protein expression levels of TRAF3, TNF-α, p65, phosphorylated p65 (p-p65), MLCK, myosin light chain (MLC), phosphorylated MLC (p-MLC), and tight junction proteins including junctional adhesion molecule-A (JAM-A), Occludin, and Claudin-1 in colonic tissues. ResultsCompared with the control group, the model group exhibited significantly increased fecal water content and serum levels of D-LA, DAO, and LPS, along with decreased protein expression levels of JAM-A, Occludin, and Claudin-1 in colonic tissues (P<0.05 or P<0.01). Additionally, in the model group, the mRNA expression levels of miR-29b-3p, TNF-α, p65, p50, and MLCK in colonic tissues were up-regulated, while the mRNA and protein expression levels of TRAF3 were down-regulated; the protein levels of TNF-α and MLCK, as well as the ratios of p-p65/p65 and p-MLC/MLC, significantly elevated (P<0.01). Compared with the model group, all treatment groups showed reduced fecal water content and serum levels of D-LA, DAO, and LPS, along with down-regulated mRNA expression levels of miR-29b-3p, TNF-α, p65, p50, and MLCK, and up-regulated TRAF3 mRNA expression in colonic tissues. Moreover, the pinaverium bromide group and high-dose Changji'an Formula group presented increased protein levels of Occludin, Claudin-1, and TRAF3, as well as decreased protein levels of TNF-α and MLCK, and reduced ratios of p-p65/p65 and p-MLC/MLC in colonic tissues (P<0.05 or P<0.01). Compared with the low-dose Changji'an Formula group, the high-dose group had lower fecal water content and serum levels of DAO and LPS (P<0.01). In comparison with the pinaverium bromide group, the high-dose Changji'an Formula group showed a significant decrease in serum DAO level (P<0.01). ConclusionsChangji'an Formula can reduce intestinal permeability and restore intestinal barrier function in IBS-D rats of liver depression and spleen deficiency syndrome by regulating the miR-29b-3p/TRAF3/NF-κB/MLCK axis.
2.Screening and verification of miRNAs differentially expressed in traumatic femoral head necrosis
Ying ZHANG ; Leilei ZHANG ; Yuna CHAI ; Ruibo SUN ; Yanan FAN ; Qiushi WEI ; Wei HE ; Huichao WANG ; Youwen LIU
Chinese Journal of Orthopaedic Trauma 2017;19(11):978-985
Objective To identify the plasma miRNAs which are differentially expressed in the patients with traumatic avascular necrosis of the femoral head by means of high-throughput screening so as to provide data for further research into the pathogenesis of traumatic avascular necrosis of the femoral head.Methods We selected 10 patients with femoral neck fracture who had been treated from January to April 2015 for traumatic avascular necrosis of the femoral head following manual reduction and internal fixation with percutaneous hollow screws (necrosis group) and another contemporary age-matched 10 patients with femoral neck fracture who had been treated in the similar way but did not develop traumatic avascular necrosis of the femoral head for over 2 years (control group).After the peripheral blood was obtained from the 20 patients,Axon GenePix 4000B chip scanner was used for peripheral blood chip screening to identify the miRNAs which were differentially expressed.QPCR technique was used to verify the top 5 miRNAs which were significantly up-regulated and the top 5 miRNAs which were significantly down-regulated.Results The Harris score for the necrosis group was significantly lower than that for the control group while the Visual Analog Scale score for the former significantly higher than that for the latter (P < 0.05).There were no significant differences between the 2 groups in general data (P > 0.05).In comparison of the 2 groups,the chip screening identified 95 miRNAs which were significantly up-regulated and 413 miRNAs which were significantly down-regulated.The "volcano" analysis screened out 147 miRNAs differentially expressed of which 35 were up-regulated genes and 112 down-regulated genes.Of the 10 genes selected,8 were found in line with the results of microarray screening:hsa-miR-93-5p,hsa-let-7i-5p,hsa-miR-320a,hsa-miR-25-3p,hsa-miR-16-2-3p,hsa-miR-122-5p,hsa-miR-4711-3p and hsa-miR-3191-5p.Conclusion This study indentified 8 differentially expressed miRNAs associated with traumatic osteonecrosis of the femoral head,providing data for further study.
3.Comparative Study on the HPLC Fingerprint of Sini Decoction Prepared by Traditional Decoction and Mod-ern Machine Decoction
Yusheng HUANG ; Hongmei TANG ; Yuanming ZHAO ; Ting WANG ; Yuna CHAI ; Yuan LIU ; Rufan ZHONG
China Pharmacy 2016;27(24):3440-3442
OBJECTIVE:To establish the HPLC fingerprints for Sini decoction and compare the differences of compositions of Sini decoction prepared by traditional decoction and modern machine decoction. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.1% phosphoric acid(gradient elution)at a flow rate of 1.0 ml/min,the detec-tion wavelength was 235 nm,column temperature was 30 ℃,and injection volume was 10 μl. The HPLC fingerprints of 10 batch-es of Sini decoction were determined with reference peak of liquiritin peaks,and common peak identification and similarity evalua-tion were conducted by using Chromatographic Fingerprint Similarity Evaluation System(2004 A edition). RESULTS:There were 18 common peaks and the similarity was no less than 0.982. According to the verification,the fingerprint of 10 batches of Sini de-coction showed good similarity with reference fingerprint,and the similarity of 10 batches of Sini decoction was high,which was prepared by the 2 methods. CONCLUSIONS:The established fingerprint is specific and stable,and can provide reference for quali-ty evaluation and control for Sini decoction;and there are no obvious differences in the main chemical compositions of Sini decoc-tion prepared by traditional decoction and modern machine decoction.
4.Cost-effectiveness Analysis of Xiyanping Injection and Ribavirin Injection for Common Type Hand-foot-mouth Disease in Children
Ting WANG ; Fen XIONG ; Hongmei TANG ; Yuna CHAI ; Yusheng HUANG ; Yuan LIU ; Rufan ZHONG
China Pharmacy 2016;27(35):4900-4903
OBJECTIVE:To compare cost-effectiveness of Xiyanping injection and Ribavirin injection in the treatment of com-mon type hand-foot-mouth disease (HFMD) in children,and to provide evidence for rational drug use in the clinic. METHODS:The literatures about Xiyanping injection in the treatment of common type HFMD in children using Ribavirin injection as control were retrieved from CNKI,VIP,Wanfang,PubMed,Cochrane library and other databases. The decision tree was established with TreeAge Pro 2011 software to conduct cost-effectiveness analysis. Tornado diagram was used to analyze sensitive factors;single fac-tor and double factors sensitivity analysis were also conducted. RESULTS & CONCLUSIONS:The total cost of Xiyanping injection and Ribavirin injection were 2 887.53 and 3 058.72 yuan,respectively. The total effective rates were 92.49% and 78.12%. Xiyan-ping injection shows cost-effectiveness advantage. The results of cost-effectiveness analysis were supported by sensitivity analysis.

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