Effect of Huangqintang on Oxidative Stress and Ferroptosis-related Indexes GSH-Px4，P53，SLC7A11 in Ulcerative Colitis Mice / 中国实验方剂学杂志

Objective:To explore the possible mechanism of Huangqintang in treating ulcerative colitis （UC）. Method:The animal model of UC was induced by dextran sodium sulfate （DSS）．The experimental animals were divided into control group， model group，Huangqintang low dose （4.55 g·kg^-1）， medium dose （9.1 g·kg^-1）， and high dose（18.2 g·kg^-1） groups. Intragastric administration was also given in the modeling process for 7 consecutive days. At the end of the 8th day， colon tissues were collected to measure colon length and mass， and calculate the colon mass index. Pathological changes were observed by hematoxylin-eosin （HE） staining. Serum iron content， superoxide dismutase （SOD）， glutathione （GSH）， catalase （CAT） and myeloperoxidase （MPO） were determined by biochemical assay. Western blot was used to detect the protein expression of glutathione peroxidase 4 （GSH-Px4）， long-chain acyl-CoA synthetase 4 （ACSL4） and ferritin heavy chain 1（FTH1）. The mRNA expression levels of tumor trotein 53 （P53） and solute carrier family 7 member 11 （SLC7A11） in colon tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:The experimental studies showed that compared with normal group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression in the model group significantly increased （<italic>P</italic><0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression in the model group significantly decreased （<italic>P</italic><0.01）. Compared with model group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression significantly decreased （<italic>P</italic><0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression significantly increased （<italic>P</italic><0.05） after the intervention of Huangqintang， and the effect was most significant in the high-dose group （<italic>P</italic><0.05）. The results of general condition， colon length， colon mass index and HE staining showed that Huangqintang could relieve clinical symptoms and histopathological changes in UC mice. Conclusion:These results indicated that Huangqintang had therapeutic effect on ulcerative colitis mice， and its mechanism might be related to inhibiting the oxidative stress and ferroptosis.

Effect of active component compound of Epimedii Folium,Astragali Radix,and Puerariae Lobatae Radix on expression of ADAM17 in HT22 cells by mediating hepcidin / 中国中药杂志

Alzheimer's disease(AD) patients in China have been surging, and the resultant medical burden and care demand have a huge impact on the development of individuals, families, and the society. The active component compound of Epimedii Folium, Astragali Radix, and Puerariae Lobatae Radix(YHG) can regulate the expression of iron metabolism-related proteins to inhibit brain iron overload and relieve hypofunction of central nervous system in AD patients. Hepcidin is an important target regulating iron metabolism. This study investigated the effect of YHG on the expression of a disintegrin and metalloprotease-17(ADAM17), a key enzyme in the hydrolysis of β amyloid precursor protein(APP) in HT22 cells, by mediating hepcidin. To be specific, HT22 cells were cultured in vitro, followed by liposome-mediated siRNA transfection to silence the expression of hepcidin. Real-time PCR and Western blot were performed to examine the silencing result and the effect of YHG on hepcidin in AD cell model. HT22 cells were randomized into 7 groups: control group, Aβ25-35 induction(Aβ) group, hepcidin-siRNA(siRNA) group, Aβ25-35 + hepcidin-siRNA(Aβ + siRNA) group, Aβ25-35+YHG(Aβ+YHG) group, hepcidin-siRNA+YHG(siRNA+YHG) group, Aβ25-35+hepcidin-siRNA+YHG(Aβ+siRNA+YHG) group. The expression of ADAM17 mRNA in cells was detected by real-time PCR, and the expression of ADAM17 protein by immunofluorescence and Western blot. Immunofluorescence showed that the ADAM17 protein expression was lower in the Aβ group, siRNA group, and Aβ+siRNA group than in the control group(P<0.05) and the expression was lower in the Aβ+siRNA group(P<0.05) and higher in the Aβ+YHG group(P<0.05) than in the Aβ group. Moreover, the ADAM17 protein expression was lower in the Aβ+siRNA group(P<0.05) and higher in the siRNA+YHG group(P< 0.05) than in the siRNA group. The expression was higher in the Aβ+siRNA+YHG group than in the Aβ+siRNA group(P<0.05). The results of Western blot and real-time PCR were consistent with those of immunofluorescence. The experiment showed that YHG induced hepcidin to up-regulate the expression of ADAM17 in AD cell model and promote the activation of non-starch metabolic pathways, which might be the internal mechanism of YHG in preventing and treating AD.

Research advance in metabolism of effective ingredients from traditional Chinese medicines by probiotics / 中国中药杂志

The pharmacological activity of active ingredients from Chinese medicine depends greatly on the microecological environment of probiotics in the human body. After effective ingredients from traditional Chinese medicines are metabolized or biotransformed by probiotics, their metabolites can increase pharmacological activity, and can be absorbed more easily to improve the bioavailability. Therefore, the combination of Chinese medicines with probiotics is the innovation point in R&D of functional food and Chinese medicines, and also a new thinking for the modernization of Chinese medicine.This review summarizes and analyses the research progress on metabolism effects of gut microbiota on Chinese medicines components, the regulating effect of effective ingredients from Chinese medicine on intestinal probiotics, the application status of probiotics in traditional Chinese medicines, and the main problems and prospects in the research and development of Chinese medicines products with probiotic, aiming to provide theoretical guidance and practical value for the fermentation engineering of Chinese herbal medicine.

Secondary metabolites of co-culture of Alternaria alternate YX-25 and Streptomyces exfoliatus YX-32 / 中草药

Objective To study the secondary metabolites of the co-culture of fungus Alternaria alternata YX-25 and Streptomyces exfoliatus YX-32 isolated from mangrove mud in Yunxiao Country of Fujian Province. Methods The compounds were purified by silica gel and Sephadex LH-20, followed by semi-preparative chromatography. The structures of the compounds were determined by MS and NMR. Results Nine compounds were isolated from the crude extract, which were identified as gliotoxin (1), 12,13-dihydroxy-fumitremorgin C (2), fumitremorgin (3), bisdethiobis(methylthio)gliotoxin (4), demethoxyfumitremorgin C (5), vermistatin (6), cyclo-(L-Trp-L-Pro) (7), 6-methoxyspirotryprostatin B (8), and citrinolactone D (9). Conclusion Nine compounds were isolated for the first time from co-culture of the fungus Alternaria alternata YX-25 and Streptomyces exfoliates YX-32, including indole diketopiperazines, polyketides, and cyclopeptides. The co-cultivation method can be as a new method for enrichment of secondary metabolites.

Cloning of Human canstatin gene and expression of its recombinant protein / 中国医学科学院学报

<p><b>OBJECTIVE</b>To clone human canstatin gene and express its recombinant protein.</p><p><b>METHODS</b>The total RNA was extracted from human placenta. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and transformed into E.coli DH5alpha through electroporation. The gene was sequenced by the Sanger Dideoxy-mediated chain-termination method, and then the canstatin cDNA was cloned into the BamHI and HindIII sites of plasmid pET-22b (+) and transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG).</p><p><b>RESULTS</b>The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting products were cloned into pUCm-T vectors, and then were transformed into E.coli DHSa. After an over night culture, both blue and white colonies were found on the agar plate. Six white colonies were selected and cut by BamHI and HindIII. The plasmids DNA in one white colony showed one band near the location of primary plasmid after digested by BamHI and two bands near the locations of primary plasmid and objective gene fragment after digested by HindIII. The cloned gene in this white colony was sequenced and demonstrated to have the same sequence as that of canstatin gene in GenBank. Then canstatin cDNA was cut down from pUCm-T with BamHI and HindIII and ligated into the vector pET-22b (+). The resultant plasmid pET-22b (+)/canstatin was then transformed into E.coli BL21. White colonies were found on LB agar plate. Seven of them were selected and their plasmids were digested with both BamHI and HindIII. After electrophoresis, all selected colonies showed two specific bands, one was found near the location of primary plasmids, and the other near that of objective gene fragment. After IPTG induction, there was a new protein band about Mr 24 000 on SDS-PAGE. As estimated by densitometry, the percentage of the expressed product over total bacterial proteins was 18.2%, 18.8%, 23.0% and 23.4%, respectively, 1, 2, 3, and 4 hours after induction.</p><p><b>CONCLUSION</b>Human canstatin gene was successfully cloned and its recombinant proteins were expressed in this study.</p>