OBJECTIVES: To study the therapeutic mechanism of Tuihuang Mixture against cholestasis. METHODS: Forty-eight Wistar rats were randomized equally into blank group, model group, ursodeoxycholic acid group and Tuihuang Mixture group. Except for those in the blank group, all the rats were given α‑naphthylisothiocyanate (ANIT) to establish rat models of cholestasis, followed by treatments with indicated drugs or distilled water. Serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL of the rats were determined, and hepatic expressions IL-1β, IL-18, FXR, NLRP3, ASC, Caspase-1 and GSDMD were detected using q-PCR, ELISA or Western blotting. Histopathological changes of the liver tissues were observed using HE staining. RESULTS: The rat models of cholestasis had significantly increased serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL with increased mRNA and protein expressions of IL-1β and IL-18, decreased protein and mRNA expressions of FXR, and increased protein expressions of NLRP3 and Caspase-1 and mRNA expressions of NLRP3, ASC, Caspase-1 and GSDMD in the liver tissue, showing also irregular arrangement of liver cells, proliferation of bile duct epithelial cells and inflammatory cells infiltration. Treatment of the rat models with Tuihuang Mixture significantly decreased serum levels of ALT, AST, ALP, γ-GT, TBA and TBIL, lowered IL-1β and IL-18 and increased FXR protein and mRNA expressions, and reduced NLRP3, ASC, Caspase-1 and GSDMD proteins and NLRP3, ASC and Caspase-1 mRNA expressions in the liver tissue. Tuihuang Mixture also significantly alleviated hepatocyte injury, bile duct epithelial cell proliferation and inflammatory cell infiltration in the liver of the rat models. CONCLUSIONS Tuihuang Mixture can effectively improve cholestasis in rats possibly by inhibiting NLRP3 inflammatosome-mediated pyroptosis via regulating FXR.
Animals ; NLR Family, Pyrin Domain-Containing 3 Protein ; Rats ; Receptors, Cytoplasmic and Nuclear/metabolism* ; Cholestasis/drug therapy* ; Rats, Wistar ; Inflammasomes/metabolism* ; 1-Naphthylisothiocyanate ; Drugs, Chinese Herbal/therapeutic use* ; Male ; Interleukin-18/metabolism* ; Caspase 1/metabolism* ; Interleukin-1beta/metabolism* ; Liver/metabolism*

ObjectiveTo investigate the effect of modified Weijingtang on the pyroptosis of RAW264.7 macrophages via the cysteinyl aspartate-specific protease-1 （Caspase-1）/gasdermin D （GSDMD） pathway. MethodLipopolysaccharide （LPS） was used to induce pyroptosis of RAW264.7 cells. The blank group was treated with the blank serum， and the intervention groups were treated with the sera containing different doses of modified Weijingtang. After 24 h， the viability of cells in different groups was examined by the cell counting kit-8 （CCK-8）. The pyroptosis and morphology of cells in each group were observed by a scanning electron microscope and a phase-contrast microscope， respectively. The mRNA and protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）， Caspase-1， and GSDMD in each group were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The levels of interleukin （IL）-18 and IL-1β in each group were measured by enzyme-linked immunosorbent assay. ResultUnder the electron microscope， RAW264.7 cells presented the best morphology and structure in the blank group and obvious pyroptosis and leakage of cell contents in the model （LPS） group. Compared with the model group， the intervention groups showed reduced pyroptosis to varying degrees， and the high-dose group had the closest cell morphology and structure to the blank group. Under the optical microscope， RAW264.7 cells were spherical in the blank group and irregular with protrusions in the model group. Compared with the model group， the intervention groups showed improved cell morphology， and the cell morphology in the group with the dose of 20% was the closest to that in the blank group. The mRNA and protein levels of NLRP3， Caspase-1， and GSDMD in the model group were higher than those in the blank group （P<0.05）. Compared with the model group， each intervention group showed down-regulated expression of the above indicators （P<0.05）. Compared with the blank group， the model group presented elevated levels of IL-18 and IL-1β （P<0.05）， which were lowered in the intervention （10%， 20%） groups （P<0.01）. ConclusionModified Weijingtang inhibits the pyroptosis of macrophages by down-regulating the Caspase-1/GSDMD pathway and reducing the release of proinflammatory cytokines.

Objective Isobaric tag for relative and absolute quantitation (iTRAQ) was applied for the quantitative analysis of serum protein in psoriasis vulgaris patients with damp heat syndrome and healthy volunteers to explore the differentially expressed proteins, thus to reveal the nature of damp heat syndrome from the proteomics level.Methods The serum from 15 psoriasis vulgaris patients with damp heat syndrome and 10 healthy volunteers was purified and treated with removal of abundance. After separation with sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and hydrolysis of peptide specific iTRAQ marker enzyme, the differentially expressed proteins of serum samples were identified by tandem mass spectrometry. Results A total of 787 proteins were identified, and 718 proteins had the functional annotation showed by gene ontology ( GO) in psoriasis vulgaris pa tients with damp heat syndrome. A total of 651 differentially expressed proteins were identified by compared with healthy volunteers ( P<0.05) , and markedly significant difference was shown in 418 proteins ( P<0.01) . Conclusion Psoriasis vulgaris patients with damp heat syndrome have differentially expressed proteins which are different from those in the healthy volunteers, but which kind of protein being specific for damp-heat psoriasis vulgaris as well as the relation between the damp heat symptoms and proteins still need to be further studied.

Objective: To study the relationship between occurrence and development of chronic periodontitis and neuropeptide substance P (SP) in gingival crevicular fluid. Methods: There were 48 patients in experimental group and 42 patients in the control group. Gingival crevicular fluid samples were collected from the patients′ intraoral chronic periodontitis inflammatory positive sites and healthy sites. Neuropeptide substance P levels in gingival crevicular fluid from two groups were measured by radioimmunoassay. Gingival index, pocket depth, attachment loss, alveolar bone resorption were recorded and their correlations with SP were analyzed. Results: SP levels of the experimental group were significantly higher than those of the control group(P<0.01). There were significant differences(P<0.01)between SP level and periodontal clinical index in slight periodontitis and the control group. There were significant correlations(P<0.05)between GCF SP content and all PD, AL, ABL but GI in slight periodontitis, and there were significant correlations in between GCF SP content and GI for both moderate periodontitis and severe periodontitis. There were significant correlations(P<0.01) between GCF SP content and PD,AL,ABL for both moderate periodontitis and severe periodontitis. Conclusion:GCF SP level is closely related to occurrence and development of chronic periodontitis.GCF SP level can reflect the degree of the damage of periodontium.Determination of patients′ GCF SP level has important significance for the patients′ condition judgement and treatment guidance.

OBJECTIVE:To establish a HS-GC method for determining methanol and ethanol in human plasma METHOD_S:An external standard method was used The stationary phase was a glass column packed with GDX-102 The carrier gas was nitrogen The column temperature was 140℃ and both the injector temperature and the detector temperature were 200℃ RE_SULTS:A good linearity was obtained in the range of 0 2～7 5g/L of methanol and ethanol The within-day RSD was less than 4% and the between-day RSD less than 5 5% The recoveries were (100 9?3 8)% for methanol,(101 5?3 4)% for et_hanol,respectively The limits of detection for both methanol and ethanol were less than 0 01g/L CONCLUSION:This method is simple,rapid and precise