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OBJECTIVE：To investigate the anti- hepatic fibrosis （HF）effects of Qiwei qinggan powder and explore its possible mechanism. METHODS ：Male Wistar rats were randomly divided into blank group ，HF model group ，Qiwei qinggan powder low-dose，medium-dose and high-dose groups [ 135，270，405 mg/（kg·d），by total amount of crude drugs] ，with 12 rats in each group. Except for blank group ，other groups were given 50％ CCl4-peanut oil solution intragastrically （2 mL/kg，twice a week ，for consecutive 8 weeks） to induce HF model. At same time ， blank group and model group were given constant volume of 0.5％ CMC-Na solution intragastrically ；administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 8 weeks. General situation of rats were observedand liver morphology was observed after last administration and hepatic indexes were detected. The contents of liverfunction indexes （ALT，AST，ALP，HYP）in serum and the expression of α-SMA in hepatic tissue were determined ， and HE and Masson staining were performed to observe the histopathology. Using the difference multiple of expression quantity as the index ，TMT technology was used to screen the differentially expressed protein in medicine group （combining the liver tissue samples of Qiwei qinggan powder groups ）and HF model group. Uniprot-GOA database and KAAS ，KEGG mapper online tools were used to analyze GO and KEGG pathway enrichment. RESULTS ：The rats in the blank group were in good health ；the liver was bright red and smooth ，the liver lobules were intact ，no degeneration and necrosis ，inflammatory cell infiltration or fibrous tissue proliferation was found. Compared with blank group ，the rats in HF model group had poor diet ，depressed spirit ，disordered and lusterless fur ；the liver was dark red or yellow with rough surface ，hard texture ，inflammatory cell infiltration ，fiber tissue destruction ，bridge connection and so on ；the hepatic index ，the contents of liver function indexes and the expression of α-SMA were increased significantly （P＜0.05）. Compared with HF model group ，above symptoms of rats were improved to different extent in different dose groups of Qiwei qinggan powder ；hepatic index in Qiwei qinggan powder low-dose group ，the content of ALP in high-dose group ，the contents of ALT，AST and HYP and the expression of α-SMA in different dose groups were decreased significantly （P＜0.05）. A total of 42 differentially expressed proteins related to HF were screened ，of which 15 were up-regulated and 27 were down-regulated in expression，including fatty acid binding protein 4（FABP4），cholesterol 7α-hydroxylase（CYP7A1）. The results of enrichment analysis showed that the differentially expressed proteins were mainly enriched in extracellular space ，blood particles and other cell parts，involving the molecular functions of oxidoreductase activity and fatty acid binding ，the biological processes of the regulation of heterotypic cell adhesion ，protein activation cascade ，as well as retinol metabolism ，arachidonic acid metabolism ，PPAR and other signal pathway. CONCLUSIONS ：Qiwei qinggan powder can reduce the hepatic index ，ALT，AST，ALP and HYP contents in serum ，down-regulate the expression of α-SMA，improve the degree of inflammation and fibrosis of liver tissue ，and have a certain protective effect on rats. The anti-HF mechanism of it involves multiple targets and signal pathways ，such as FABP 4， CYP7A1 and PPAR.
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<p><b>OBJECTIVE</b>To develop a green and rapid method for extraction of lobetyolin from C. pilosula.</p><p><b>METHOD</b>Extraction of lobetyolin from C. pilosula with supercritical carbon dioxide in the presence of ethanol was studied. The effects of pressure, temperature, volume of cosolvent and extraction time on efficiency and their interactive relationships were discussed, based on central composite design and response surface methodology (RSM).</p><p><b>RESULT</b>The key effect factor was volume of cosolvent. The extraction yield of lobetyolin was 0.078 6 mg x g(-1) when C. pilosula (40-60 mesh) was extracted at 30 MPa, 60 degrees C and 2 L x min(-1) (as CO2 in normal pressure and temperature) for 100 minutes with supercritical CO2 and 1 mL x min(-1) ethanol as dynamic cosolvent.</p><p><b>CONCLUSION</b>This result is better than that obtained from traditional method. Therefore, the optimized process is valuable for extraction of lobetyolin from C. pilosula.</p>
Subject(s)
Carbon Dioxide , Chemistry , Chemical Fractionation , Methods , Codonopsis , Chemistry , Drugs, Chinese Herbal , Chemistry , Ethanol , Chemistry , Polyacetylene Polymer , ChemistryABSTRACT
Objective To investigate the oxidative mechanism of homocysteine ( Hey) -induced apoptosis in endothelial progenitor cells( EPCs). Methods Total mononuclear cells were isolated from mouse bone marrow by Ficoll density gradient centrifugation and were cultured in vitro for 7 d. Adherent cells were harvested and identified by fluorescence microscopy. EPCs were cultured with Hey (0, 50, 100 and 500 μmol/L) for 12, 24 and 48 h, or pretreated with NAC (1 mmol/L), DPI( 10 μmol/L) or SB203580 (10 μmol/L) for 30 min, then cultured with 500 μmol/L Hey for 24 h. Apoptosis was detected by Annexin-V/PI flow cytometry, levels of reactive oxygen species (ROS) in cells were measured using H2DCF-DA as a fluorescence probe, NADPH oxidases were evaluated with lucigenin-enhanced chemilumine9cence, and NO in the supernatant was determined by nitrate reductase assay. Results Hey induced EPCs apoptosis, ROS accumulation, NADPH oxidase activation and decrease of NO in a time-dose dependent manner( P <0.05 or P < 0.01). Pretreatment with NAC, DPI and SB203580 could inhibit these effects (P < 0.05 or P < 0.01). Conclusion Hey could activate NADPH oxidase, induce ROS increase and NO decrease, and activate p38MAPK to enhance EPCs apoptosis.
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OBJECTIVE:To make clear the antimicrobials used for inpatients in our hospital from September 1999 to March 2000 METHODS:We sampled the records of discharged patients at the ratio of 1/3 of sickbeds in our hospital,and filled in "the questionnaire of antimicrobials used in clinical department" item by item,finally,we carried out comprehensive analysis RESU_LTS:More than half was administration of single and basic antimicrobial Combined use of antimicrobials for severe and mixed infections was rational The rate of using antimicrobials was high(79 64%),especially in preventing perioperative infections,reached nearly 100% The starting-point was high in choice of drug and the newly-developed,special and expensive drugs occupied the front place in order of consumption Combined use of drugs was unduly prevalent Most of treatment depended on experience The rate of bacterial drug resistance test was only performed in 10% of patients CONCLUSION:Rational use of drugs is the main trend,however,the measure directed against existing problem should be worked out
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Objective: To compare nephrotoxicity and ototoxicity of gentamycin administered in single dose or multiple dose daily in guinea pigs. Methods: Thirty two male guinea pigs were divided into physiological saline control, single dose group daily (gentamycin, 120 mg/kg, 1/d) and multiple dose group daily (gentamycin, 60 mg/kg, 2/d). The physiopathology of renal and cochlea in guinea pigs were examined using auditory brainstem response (ABR), SC sound irritation and electron microscope. The gentamycin concentrations in serum and in perilymph were monitored by fluorescene polarization immunoassay (FPIA). Results: (1) Compared with control group, both gentamycin single and mulitiple daily doses injuried kidney and cochlea to some extent.The injury of multiple dose groups were worse than that the single dose groups ( P