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AIM:Using bioinformatics analysis and experiment validation to explore the differential expres-sion genes related to abnormal lipid metabolism in hepatocellular carcinoma(HCC)and the molecular mechanism of pachymaran affecting pyroptosis through squalene epoxidase(SQLE)/nucleotide-binding oligomerization domain-like re-ceptor protein 3(NLRP3)/gasdermin D(GSDMD)signaling pathway.METHODS:(1)The GEO,GSEA,DAVID,STRING and GEPIA databases were employed to screen abnormal lipid metabolism-related differentially expressed genes in HCC.(2)The tumor tissues from HCC patients(n=9)were collected to verify the differential expression of SQLE.(3)The inhibitory effect of pachymaran on the viability of human HCC cell line HepG2 was measured by CCK-8 assay.(4)The HepG2 cells were divided into control group and pachymaran(800 mg/L)group.The cell migration was analyzed by wound-healing assay,and RT-qPCR was used to measure SQLE mRNA expression.(5)The HepG2 cells with overexpres-sion of SQLE(OE-SQLE)were divided into 5 groups as follows:control group,overexpression negative control(OE-NC)group,OE-SQLE group,OE-NC+pachymaran group,and OE-SQLE+pachymaran group.The mRNA and protein expres-sion levels of SQLE and pyroptosis-related factors were determined by RT-qPCR and Western blot.Colorimetric method and ELISA were used to measure lactate dehydrogenase(LDH),interleukin-1β(IL-1β)and IL-18 levels.The necrosis of HepG2 cells was analyzed by flow cytometry.RESULTS:The SQLE gene was screened through bioinformatics analysis,and its mRNA expression was significantly increased in tumor tissues from HCC patients(P＜0.01).In cell experiments,treatment with 800 mg/L pachymaran for 48 h had a significant inhibitory effect on HepG2 cell viability,and the expres-sion of SQLE mRNA was reduced(P＜0.01).After overexpression of SQLE,the mRNA and protein levels of pyroptosis-re-lated factors,necrotic rate,and LDH,IL-1β and IL-18 levels were significantly decreased(P＜0.05).After treatment with pachymaran,the above indicators were significantly increased(P＜0.05).CONCLUSION:The SQLE is abnormal-ly highly expressed in HCC,and pachymaran can affect the growth of HCC cells by activating the NLRP3/GSDMD pyropto-sis pathway through SQLE.
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Kaixin Powder, an ancient Chinese herbal decoction and is composed of Ginseng Radix et Rhizoma, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria Cocos. Its main active chemical components are ginseng ginsenosides, polygalae saponins, oligosaccharide esters, asarone, poria acid, and pachymaran, which have a variety of pharmacological effects such as anti-depression, anti-dementia, improving learning and memory, and anti-fatigue. This article mainly reviewed the literature related to Kaixin Powder published in domestic and foreign research journals, summarized the research on the pharmacological activity and their mechanisms of Kaixin Powder, and prospected the potential clinical application in the treatment of mental illness.
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Guizhi Fuling Prescription is a classic prescription for promoting blood activation and resolving stasis. It has the functions of improving blood rheology, regulating endocrine, anti-inflammation, anti-cancer, and analgesia. In ancient times, it was used to treat women’s bleeding during pregnancy, abdominal pain, and postpartum lochia. It is also often used to treat gynecological diseases in modern clinics, such as uterine fibroids, ovarian cysts, dysmenorrhea, endometriosis and so on. This paper reviewed the research progress of chemical constituents, pharmacological activities, clinical application, and quality control of Guizhi Fuling Prescription in recent years. On the basis of this, the quality markers of Guizhi Fuling Prescription are predicted and analyzed based on the “five principles” of quality markers of Chinese materia medica in order to provide reference for the study of quality control of Guizhi Fuling Prescription.
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Aim To determine the neuroprotective effect of sulfated pachymaran (SP)on MPTP-induced mouse model.Method ICR mice were randomly di-vided into control group,MPTP group and SP treatment group (50,1 00,1 50 mg·kg -1 ,ip).After 1 7 days, the activities of endogenous antioxidants (SOD,GSH-Px,CAT),antisuperoxide anion,hydrogen peroxide and MDA content in the midbrain and cortex were as-sayed.Results The results proved that SP significant-ly reduced the content of MDA and H2 O2 ,regulated the activities of antioxidant enzyme and increased the activity of antisuperoxide anion.Conclusion All these effects indicate that SP is a potential neuroprotective a-gent and its neuroprotective effects are achieved in the MPTP mouse model.
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Objective: To establish a method for the determination of total glycosides in Chinese materia medica (CMM)-differential anthrone-sugar hydrazone colorimetry. Methods: The content of total glycosides (including sugar) in CMM was determined by anthrone colorimetry, and then sugar hydrazone-sulfuric acid colorimetric method was used to determine the content of reducing sugars. The content of total glycosides came from the two subtraction. Results: Anthrone colorimetric reaction was completed within 6 h, detection wavelength was 620 nm, linear range was 80-180 μg/mL, correlation coefficient R2 = 0.994, and recovery was 99.2%, RSD was 1.35%. Sugar hydrazone-sulfuric acid color reaction method was completed within 4 h, detection wavelength was 412 nm, linear range was 2-12 mg/mL, correlation coefficient R2 = 0.991, and recovery was 99.5%, RSD was 1.98%. The total glycosides contents in five Chinese patent medicines such as Buyang Huanwu Decoction, Liuwei Dihuang Pill, Andrographis Tablet, Astragalus Injection, and Pachymaran Oral Liquid were 15.1, 26.0, 6.68 mg/g, and 21.3, 16.9 mg/mL. Conclusion: This method is firstly established for the quantitative determination of total glycosides in CMM, and the ingredient group analysis method is established by anthrone colorimetry and sugar hydrazone-sulfuric acid colorimetric method. The methods have better accuracy, and can be used for the determination of total glycosides ingredients group of CMM.
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Objective To study the effect of pine root liquid on the fermentation Poria mycelia yield and content of water-soluble polysaccharide, and difference between fermentation Poria mycelium and the natural Poria. Methods Poria mycelia was cultured with pine root solution and non-fermented liquid, the production of water-soluble polysaccharide content was compared. The content of water-soluble polysaccharide of fermentation mycelia and natural Poria was also compared. Results The production of fermentation Poria mycelia cultured with pine root liquid increased slightly, but water-soluble polysaccharide content decreased. Water-soluble polysaccharide content in Poria mycelia was significantly higher than that in natural Poria. Conclusion Pine root solution has different effect on fermentation Poria mycelia yield and polysaccharide content. Water-soluble polysaccharide content in Poria fermentation mycelia was higher than natural Poria.
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Objective To study the antileukemia effect of Carboxymethyl pachymaran (CMP) on p388 leukemia mice and its molecular mechanism. Methods After establishing leukemia normal model, they were randomly divided into groups and treated at the same time. FCM and mRNA in situ hybridization(ISH) and S- P assay immunocytochemistry were used for apoptosis and the expression of bcl- 2 mRNA and bcl- 2 protein in peripheral blood lymphocyte samples. Results CMP could prolong the span time of tumor- bearing mice 35.88 %, it was significant for CMP group compared with model group( P