Questo lavoro è stato sostenuto dalla National Science Foundation della Cina (81573992). Ringraziamo Emily K. Lo e Kathleen DiNapoli per la loro assistenza per la modifica del linguaggio.

Il protocollo segue le linee guida per la cura degli animali dell&#39;Università di Medicina tradizionale cinese di Shanghai ed è approvato dall&#39;esperimento animale Comitato di etica degli animali di Shanghai. 1. Protocollo I: preparazione di EXD <ol><li> Calcolo <ol><li> Amministrare l&#39;EXD ad un gruppo di trattamento di 10 minicenne femminile (CNR) (0.02 kg / mouse), 5 giorni alla settimana per 12 settimane. NOTA: sono necessari complessivamente 1.702 EXD. Poiché gli EXD sono misurati in incrementi discreti, in pratica reale, amministrare 2 EXDs. </li><li> Calcolare il volume di EXD applicato per mouse (V = (0,1418 EXD / kg • giorno) * (0,02 kg / mouse) * 50 mL / EXD = 0,14 mL / mouse • giorno). </li></ol></li><li> ammollo <ol><li> Mettete tutte le materie prime di due EXD ( cioè, Curculigo orchioides Gaertn (18 g), Herbaa Epimedii (18 g), Radix Morindae Officinalis (18 g), RAdix Angelicae Sinensis (18 g), Cortex Phellodendri (12 g) e Rhizoma Anemarrhenae (12 g), 96 g in totale) in un contenitore con coperchio. NOTA: Si consiglia un contenitore in ceramica. </li><li> Macerare le erbe crude con 500 ml di acqua distillata per 1 h; L&#39;acqua dovrebbe coprire le erbe di circa un pollice. Lasciate che tutte le erbe abbassino completamente. </li></ol></li><li> Primo decotto d&#39;acqua <ol><li> Scaldare le erbe utilizzando un fornello a gas o un fornello ad induzione ad alta potenza fino a quando l&#39;acqua bolle (circa 5 - 10 min; il tempo dipende dalla potenza termica, dal contenitore e dalla quantità di erbe e acqua). Abbassare il potere a un minimo di zucchero per 2 ore. </li></ol></li><li> Prima filtrazione <ol><li> Coprire un bicchiere di vetro da 500 ml con carta filtro regolare o con garza e cotone. Spegnere con cautela il decotto nel bicchiere attraverso il filtro. Lasciare le erbe nel contenitore. </li><li> Ritorna il residuo vegetale residuo oN la carta filtrante nel contenitore. </li></ol></li><li> Seconda decotto dell&#39;acqua <ol><li> Aggiungere l&#39;acqua distillata al contenitore con le erbe; Lasciate che copri le erbe di circa un pollice. </li><li> Ripetere il passo 1.3. </li></ol></li><li> Seconda filtrazione <ol><li> Ripetere i passaggi 1.4.1 e 1.4.2. </li><li> Versare la seconda decozione nello stesso bicchiere. Mescolare insieme la prima e la seconda decotto. </li></ol></li><li> Concentrazione <ol><li> Mettere il bicchiere su un fornello a gas con la garza di legare senza amianto tra di loro, scaldare la decozione usando un basilico basso e mescolare lentamente e continuamente con una barra di vetro. </li><li> Quando la decozione si riduce a 200 ml, trasferirla in un bicchiere da 500 ml. </li><li> Ripetere il passo 1.7.1 fino a decottare fino a 100 ml. </li></ol></li><li> Stoccaggio e amministrazione <ol><li> Trasferire la decotto concentrata in una bottiglia di vetro sterile. Lasciare raffreddare a temperatura ambienteRe (RT). Conservare a 4 ° C se impiegato entro una settimana oa -70 ° C per la conservazione a lungo termine. </li><li> Utilizzando un ago di gavaggio, somministrare 0,14 ml / mouse del decotto a topi ovariectomizzati (OVX) una volta al giorno, 5 giorni alla settimana per 12 settimane. </li></ol></li></ol> 2. Protocollo II: Preparazione del siero contenente EXD <ol><li> Calcolo <ol><li> Calcolare la dose equivalente del ratto (ROSSO) usando 0,15 kg come peso corporeo di ciascun ratto (1 mese): dB = (1/60) * (90/100) * (60 / 0,15) 1/3 = 0,111 EXD / kg / die. NOTA: Per la preparazione di 100 ml di terreno di coltura (10% di siero contenente EXD) è necessario un totale di 10 ml di siero contenente EXD. Ogni ratto dovrebbe fornire 2 ml di siero. Quindi, 6 ratti (una perdita del 20% è presa in considerazione) sono necessari per l&#39;associazione EXD una volta al giorno per 3 giorni. Il numero di EXD è: (0,111 EXD / kg • giorno) * (0,15 kg / ratto) * (6 ratti) * (3 giorni) = 0,300 EXD. Anche in questo caso, poiché gli EXD sono misurati in incrementi discreti,Utilizzare 1 EXD. </li><li> Calcolare il volume di EXD applicato per ratto (v = (0.111 EXD / kg • giorno) * (0.15 kg / ratto) * (50 mL / EXD) = 0.83 mL / ratto • giorno). </li></ol></li><li> ammollo <ol><li> Mettere le materie prime per 1 EXD in un contenitore con coperchio. Macerare le erbe crude in acqua distillata per 1 h; L&#39;acqua dovrebbe coprire le erbe di circa un pollice. Lasciate che tutte le erbe abbassino completamente. </li></ol></li><li> Primo decotto d&#39;acqua <ol><li> Scaldare le erbe utilizzando un fornello ad induzione ad alta potenza fino a quando l&#39;acqua bolle (circa 5 - 10 min; il periodo di tempo dipende dalla potenza termica, dal contenitore e dalla quantità di erbe e acqua). Spegnere l&#39;apparecchio fino a raggiungere un minimo di 2 ore. </li></ol></li><li> Prima filtrazione <ol><li> Coprire un bicchiere di vetro da 500 ml con carta filtro regolare o con garza e cotone. Spegnere con cautela il decotto nel bicchiere attraverso il filtro. Lasciare le erbe nel contenitore. </li><li&gt; Rimettere il residuo vegetale sulla carta filtrante nel contenitore. </li></ol></li><li> Seconda decotto dell&#39;acqua <ol><li> Aggiungere l&#39;acqua distillata al contenitore con le erbe; Lasciate che copri le erbe di circa un pollice. </li><li> Ripetere il passo 2.4. </li></ol></li><li> Seconda filtrazione <ol><li> Ripetere i passaggi 2.5.1 e 2.5.2. </li><li> Versare la seconda decottazione nello stesso bicchiere come nel punto 2.4.1 e mescolare insieme la prima e la seconda decotto. </li></ol></li><li> Concentrazione <ol><li> Mettere il bicchiere su un fornello a gas con la garza di legare senza amianto tra di loro, scaldare la decozione usando un basso affumicatoio e mescolare lentamente e continuamente con una barra di vetro. </li><li> Quando la decozione si riduce a 100 ml, trasferirla in un bicchiere da 100 ml. </li><li> Ripetere il passo 2.7.1 fino a decottare fino a 50 ml. </li></ol></li><li> Stoccaggio e amministrazione <ol><li> Trasferire la decottazione concentrata in uno sterBottiglia di vetro del ile. Lasciare raffreddare a RT. Conservare a 4 ° C se impiegato entro una settimana oa -70 ° C per la conservazione a lungo termine. </li><li> Intragastricamente somministrare 0,83 ml / ratto una volta al giorno per 3 giorni. </li></ol></li><li> Preparazione del siero contenente EXD <ol><li> Anestetizzare i ratti mediante iniezione intraperitoneale di 300 mL / 100 g di 80 mg / kg di ketamina e 10 mg / kg di xilazina a 1 ora dopo l&#39;ultima somministrazione di EXD. Confermare l&#39;anestesia appropriata con il pizzico del piede. </li><li> Incise la pelle e il peritoneo del ratto dall&#39;addome al fondo del torace usando un bisturi (o forbici diritte). Creare un&#39;incisione di lunghezza e profondità rispettivamente di circa 5 cm e 0,5 cm. Spostare la viscera addominale a sinistra usando carta tissue. </li><li> Usando carta tissue, rimuovere il tessuto connettivo dell&#39;arto addominale per esporre chiaramente il vaso. </li><li> Tirate lentamente il sangue dall&#39;arto addominale usando una siringa da 10 ml di 22 gauge. TrasferirloUn tubo sterile da 15 ml dopo aver rimosso l&#39;ago; Un ratto può produrre 8-10 ml di sangue. NOTA: il ratto dovrebbe essere vivo ( cioè, con l&#39;aorta pulsata) quando inizia il disegno del sangue e morta dopo che il disegno viene completato. </li><li> Mescolare il sangue in posizione verticale per 30-60 minuti a camera RT. Centrifugare a 500-600 xg per 20 min. Collocare con cura tutto il supernatante (siero) in un tubo sterile da 50 ml e mescolare insieme tutto il siero (da diversi animali) insieme. </li><li> Effettuare l&#39;inattivazione del calore incubando il siero in un bagno d&#39;acqua di 56 ° C per 30 min. Filtrare il siero utilizzando un filtro a siringa con una membrana idrofila di polietersolfone di pori 0,22 μm. Usare il siero fresco o conservarlo a -20 ° C. </li></ol></li><li> Controllo della preparazione sierica contenente farmaci (utilizzata nel gruppo di controllo) <ol><li> Amministrare ogni ratto con lo stesso volume di soluzione salina (0,83 ml) una volta al giorno per 3 giorni. Gli altri passaggi sono gli stessi del passaggio 2.9.</li></ol></li><li> Applicazione <ol><li> Aggiungere 10 ml di siero contenente EXD o di siero contenente droga ad ogni 100 ml di mezzo, più 1% di penicillina-streptomicina (PS) 23 . </li></ol></li></ol>

<ol><li>Zhang, L. G., Ouyang, X. W., Wu, T. T., Ni, L. J., Shi, W. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Quantitative+evaluation+of+in+vitro+effects+and+interactions+of+active+fractions+in+a+Chinese+medicinal+formula+%28Yaotongning+Capsule%29+on+rat+chondrocytes%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">Quantitative evaluation of in vitro effects and interactions of active fractions in a Chinese medicinal formula (Yaotongning Capsule) on rat chondrocytes.</a> J Ethnopharmacol. 155 (3), 1424-1432 (2014).</li>
<li>Ogawa, Y., Fujii, Y., Sugiyama, R., Konishi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+role+of+the+seven+crude+drug+components+in+the+sleep-promoting+effect+of+Yokukansan%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">The role of the seven crude drug components in the sleep-promoting effect of Yokukansan.</a> J Ethnopharmacol. 177, 19-27 (2016).</li>
<li>Kim, J. H., Doh, E. J., Lee, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evaluation+of+Medicinal+Categorization+of+Atractylodes+japonica+Koidz.+by+Using+Internal+Transcribed+Spacer+Sequencing+Analysis+and+HPLC+Fingerprinting+Combined+with+Statistical+Tools%5BTitle%5D)+AND+%22Evid+Based+Complement+Alternat+Med%22%5BJournal%5D)">Evaluation of Medicinal Categorization of Atractylodes japonica Koidz. by Using Internal Transcribed Spacer Sequencing Analysis and HPLC Fingerprinting Combined with Statistical Tools.</a> Evid Based Complement Alternat Med. 2016, 2926819(2016).</li>
<li>Sheridan, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+potential+of+metabolic+fingerprinting+as+a+tool+for+the+modernisation+of+TCM+preparations%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">The potential of metabolic fingerprinting as a tool for the modernisation of TCM preparations.</a> J Ethnopharmacol. 140 (3), 482-491 (2012).</li>
<li>Planas, G. M., Kucacute, J. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/pubmed/17744732">Contraceptive Properties of Stevia rebaudiana.</a> Science. 162 (3857), 1007(1968).</li>
<li>Tashino, S. "Serum pharmacology" and "serum pharmaceutical chemistry": from pharmacology of Chinese traditional medicines to start a new measurement of drug concentration in blood. Ther Drug Monit Res. 5, 54-64 (1988).</li>
<li>Cao, Y., Liu, F., Huang, Z., Zhang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protective+effects+of+Guanxin+Shutong+capsule+drug-containing+serum+on+tumor+necrosis+factor-alpha-induced+endothelial+dysfunction+through+nicotinamide+adenine+dinucleotide+phosphate+oxidase+and+the+nitric+oxide+pathway%5BTitle%5D)+AND+%22Exp+Ther+Med%22%5BJournal%5D)">Protective effects of Guanxin Shutong capsule drug-containing serum on tumor necrosis factor-alpha-induced endothelial dysfunction through nicotinamide adenine dinucleotide phosphate oxidase and the nitric oxide pathway.</a> Exp Ther Med. 8 (3), 998-1004 (2014).</li>
<li>Fu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ex+Vivo+Stromal+Cell-Derived+Factor+1-Mediated+Differentiation+of+Mouse+Bone+Marrow+Mesenchymal+Stem+Cells+into+Hepatocytes+Is+Enhanced+by+Chinese+Medicine+Yiguanjian+Drug-Containing+Serum%5BTitle%5D)+AND+%22Evid+Based+Complement+Alternat+Med%22%5BJournal%5D)">Ex Vivo Stromal Cell-Derived Factor 1-Mediated Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells into Hepatocytes Is Enhanced by Chinese Medicine Yiguanjian Drug-Containing Serum.</a> Evid Based Complement Alternat Med. 2016, 7380439(2016).</li>
<li>Chen, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Application+of+serum+pharmacology+in+evaluating+the+antitumor+effect+of+Fuzheng+Yiliu+Decoction+from+Chinese+Medicine%5BTitle%5D)+AND+%22Chin+J+Integr+Med%22%5BJournal%5D)">Application of serum pharmacology in evaluating the antitumor effect of Fuzheng Yiliu Decoction from Chinese Medicine.</a> Chin J Integr Med. 20 (6), 450-455 (2014).</li>
<li>Sze, S. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+mechanism%3A+Erxian+Decoction%2C+a+Chinese+medicine+formula%2C+for+relieving+menopausal+syndrome%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">A novel mechanism: Erxian Decoction, a Chinese medicine formula, for relieving menopausal syndrome.</a> J Ethnopharmacol. 123 (1), 27-33 (2009).</li>
<li>Sze, S. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+Erxian+decoction%2C+a+Chinese+medicinal+formulation%2C+on+serum+lipid+profile+in+a+rat+model+of+menopause%5BTitle%5D)+AND+%22Chin+Med%22%5BJournal%5D)">Effects of Erxian decoction, a Chinese medicinal formulation, on serum lipid profile in a rat model of menopause.</a> Chin Med. 6, 40(2011).</li>
<li>Zhong, L. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+randomized%2C+double-blind%2C+controlled+trial+of+a+Chinese+herbal+formula+%28Er-Xian+decoction%29+for+menopausal+symptoms+in+Hong+Kong+perimenopausal+women%5BTitle%5D)+AND+%22Menopause%22%5BJournal%5D)">A randomized, double-blind, controlled trial of a Chinese herbal formula (Er-Xian decoction) for menopausal symptoms in Hong Kong perimenopausal women.</a> Menopause. 20 (7), 767-776 (2013).</li>
<li>Wang, S. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Steroidogenic+effect+of+Erxian+decoction+for+relieving+menopause+via+the+p-Akt%2FPKB+pathway+in+vitro+and+in+vivo%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">Steroidogenic effect of Erxian decoction for relieving menopause via the p-Akt/PKB pathway in vitro and in vivo.</a> J Ethnopharmacol. , (2016).</li>
<li>Lin, L., Wu, S., Tang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((%5BClinical+observation+and+experimental+study+of+the+treatment+of+aplastic+anemia+by+warming+and+tonifying+the+spleen+and+kidney%5D%5BTitle%5D)+AND+%22Zhong+Xi+Yi+Jie+He+Za+Zhi%22%5BJournal%5D)">[Clinical observation and experimental study of the treatment of aplastic anemia by warming and tonifying the spleen and kidney].</a> Zhong Xi Yi Jie He Za Zhi. 10 (5), 272-274 (1990).</li>
<li>Nian, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antiosteoporotic+activity+of+Er-Xian+Decoction%2C+a+traditional+Chinese+herbal+formula%2C+in+ovariectomized+rats%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">Antiosteoporotic activity of Er-Xian Decoction, a traditional Chinese herbal formula, in ovariectomized rats.</a> J Ethnopharmacol. 108 (1), 96-102 (2006).</li>
<li>Qin, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antiosteoporotic+chemical+constituents+from+Er-Xian+Decoction%2C+a+traditional+Chinese+herbal+formula%5BTitle%5D)+AND+%22J+Ethnopharmacol%22%5BJournal%5D)">Antiosteoporotic chemical constituents from Er-Xian Decoction, a traditional Chinese herbal formula.</a> J Ethnopharmacol. 118 (2), 271-279 (2008).</li>
<li>Xue, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+HNMR-based+metabonomics+study+of+postmenopausal+osteoporosis+and+intervention+effects+of+Er-Xian+Decoction+in+ovariectomized+rats%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">A HNMR-based metabonomics study of postmenopausal osteoporosis and intervention effects of Er-Xian Decoction in ovariectomized rats.</a> Int J Mol Sci. 12 (11), 7635-7651 (2011).</li>
<li>Lu, X. N., Xu, X. R., Lin, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((%5BClinical+observation+of+bushen+er%27xian+decoction+in+treating+premature+ovarian+failure%5D%5BTitle%5D)+AND+%22Zhongguo+Zhong+Xi+Yi+Jie+He+Za+Zhi%22%5BJournal%5D)">[Clinical observation of bushen er'xian decoction in treating premature ovarian failure].</a> Zhongguo Zhong Xi Yi Jie He Za Zhi. 28 (7), 594-596 (2008).</li>
<li>Yu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Anti-angiogenic+activity+of+Erxian+Decoction%2C+a+traditional+Chinese+herbal+formula%2C+in+zebrafish%5BTitle%5D)+AND+%22Biol+Pharm+Bull%22%5BJournal%5D)">Anti-angiogenic activity of Erxian Decoction, a traditional Chinese herbal formula, in zebrafish.</a> Biol Pharm Bull. 35 (12), 2119-2127 (2012).</li>
<li>Chu, E. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+in+vitro+and+in+vivo+investigation+of+the+antimetastatic+effects+of+a+Chinese+medicinal+decoction%2C+erxian+decoction%2C+on+human+ovarian+cancer+models%5BTitle%5D)+AND+%22Integr+Cancer+Ther%22%5BJournal%5D)">An in vitro and in vivo investigation of the antimetastatic effects of a Chinese medicinal decoction, erxian decoction, on human ovarian cancer models.</a> Integr Cancer Ther. 12 (4), 336-346 (2013).</li>
<li>Zhu, Z., Li, L., Jin, X., Fang, J., Zhang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Er-Xian+Decoction%2C+a+traditional+Chinese+herbal+formula%2C+intervening+early+in+hypothalamic-pituitary+axis+of+male+rats+with+delayed+puberty%5BTitle%5D)+AND+%22Pharmacogn+Mag%22%5BJournal%5D)">Er-Xian Decoction, a traditional Chinese herbal formula, intervening early in hypothalamic-pituitary axis of male rats with delayed puberty.</a> Pharmacogn Mag. 10 (40), 517-521 (2014).</li>
<li>Miao, M. S. Experimental animals and technology. 1997, Chinese Modical Publisher. Beijing. 145(1997).</li>
<li>Soleimani, M., Nadri, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+protocol+for+isolation+and+culture+of+mesenchymal+stem+cells+from+mouse+bone+marrow%5BTitle%5D)+AND+%22Nat+Protoc%22%5BJournal%5D)">A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow.</a> Nat Protoc. 4 (1), 102-106 (2009).</li>
<li>Lu, S., Huang, J., Wang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Er-Xian+Decoction+Stimulates+Osteoblastic+Differentiation+of+Bone+Mesenchymal+Stem+Cells+in+Ovariectomized+Mice+and+Its+Gene+Profile+Analysis%5BTitle%5D)+AND+%22Stem+Cells+International%22%5BJournal%5D)">Er-Xian Decoction Stimulates Osteoblastic Differentiation of Bone Mesenchymal Stem Cells in Ovariectomized Mice and Its Gene Profile Analysis.</a> Stem Cells International. 2016, 4079210(2016).</li>
<li>Turner, P. V., Brabb, T., Pekow, C., Vasbinder, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Administration+of+substances+to+laboratory+animals%3A+routes+of+administration+and+factors+to+consider%5BTitle%5D)+AND+%22J+Am+Assoc+Lab+Anim+Sci%22%5BJournal%5D)">Administration of substances to laboratory animals: routes of administration and factors to consider.</a> J Am Assoc Lab Anim Sci. 50 (5), 600-613 (2011).</li>
<li>Wu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Simultaneous+determination+of+seven+flavonoids+in+dog+plasma+by+ultra-performance+liquid+chromatography-tandem+mass+spectrometry+and+its+application+to+a+bioequivalence+study+of+bioactive+components+in+Herba+Epimedii+and+Er-Xian+Decoction%5BTitle%5D)+AND+%22J+Pharm+Biomed+Anal%22%5BJournal%5D)">Simultaneous determination of seven flavonoids in dog plasma by ultra-performance liquid chromatography-tandem mass spectrometry and its application to a bioequivalence study of bioactive components in Herba Epimedii and Er-Xian Decoction.</a> J Pharm Biomed Anal. 54 (1), 186-191 (2011).</li>
<li>Hu, Y. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Identification+of+the+major+chemical+constituents+and+their+metabolites+in+rat+plasma+and+various+organs+after+oral+administration+of+effective+Erxian+Decoction+%28EXD%29+fraction+by+liquid+chromatography-mass+spectrometry%5BTitle%5D)+AND+%22Biomed+Chromatogr%22%5BJournal%5D)">Identification of the major chemical constituents and their metabolites in rat plasma and various organs after oral administration of effective Erxian Decoction (EXD) fraction by liquid chromatography-mass spectrometry.</a> Biomed Chromatogr. 24 (5), 479-489 (2010).</li>
<li>Xue, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+and+interaction+of+icariin%2C+curculigoside%2C+and+berberine+in+er-xian+decoction%2C+a+traditional+chinese+medicinal+formula%2C+on+osteoclastic+bone+resorption%5BTitle%5D)+AND+%22Evid+Based+Complement+Alternat+Med%22%5BJournal%5D)">Effects and interaction of icariin, curculigoside, and berberine in er-xian decoction, a traditional chinese medicinal formula, on osteoclastic bone resorption.</a> Evid Based Complement Alternat Med. , (2012).</li>
<li>Nair, A. B., Jacob, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+simple+practice+guide+for+dose+conversion+between+animals+and+human%5BTitle%5D)+AND+%22J+Basic+Clin+Pharm%22%5BJournal%5D)">A simple practice guide for dose conversion between animals and human.</a> J Basic Clin Pharm. 7 (2), 27-31 (2016).</li>
<li>Xu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protective+effect+of+the+traditional+Chinese+medicine+xuesaitong+on+intestinal+ischemia-reperfusion+injury+in+rats%5BTitle%5D)+AND+%22Int+J+Clin+Exp+Med%22%5BJournal%5D)">Protective effect of the traditional Chinese medicine xuesaitong on intestinal ischemia-reperfusion injury in rats.</a> Int J Clin Exp Med. 8 (2), 1768-1779 (2015).</li>
<li>Li, Z., Wang, J. On the methods for Chinese herbs serum pharmcology. Zhong Guo Zhong Yi Yao Xing Xi Za Zhi. 9 (2), 5-6 (2002).</li>
<li>Jiang, Y. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+chinese+herbal+drug-containing+serum+for+activating-blood+and+dispelling-toxin+on+ox-LDL-induced+inflammatory+factors%27+expression+in+endothelial+cells%5BTitle%5D)+AND+%22Chin+J+Integr+Med%22%5BJournal%5D)">Effect of chinese herbal drug-containing serum for activating-blood and dispelling-toxin on ox-LDL-induced inflammatory factors' expression in endothelial cells.</a> Chin J Integr Med. 18 (1), 30-33 (2012).</li>
<li>Guo, C. Y., Ma, X. J., Liu, Q., Yin, H. J., Shi, D. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((%5BEffect+of+Chinese+herbal+drug-containing+serum+for+activating+blood%2C+activating+blood+and+dispelling+toxin+on+TNF-alpha-induced+adherence+between+endothelial+cells+and+neutrophils+and+the+expression+of+MAPK+pathway%5D%5BTitle%5D)+AND+%22Zhongguo+Zhong+Xi+Yi+Jie+He+Za+Zhi%22%5BJournal%5D)">[Effect of Chinese herbal drug-containing serum for activating blood, activating blood and dispelling toxin on TNF-alpha-induced adherence between endothelial cells and neutrophils and the expression of MAPK pathway].</a> Zhongguo Zhong Xi Yi Jie He Za Zhi. 35 (2), 204-209 (2015).</li>
<li>Li, Y., Xia, J. Y., Chen, W., Deng, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+Ling+Qi+Juan+Gan+capsule+drug-containing+serum+on+PDGF-induced+proliferation+and+JAK%2FSTAT+signaling+of+HSC-T6+cells%5D%5BTitle%5D)+AND+%22Zhonghua+Gan+Zang+Bing+Za+Zhi%22%5BJournal%5D)">Effects of Ling Qi Juan Gan capsule drug-containing serum on PDGF-induced proliferation and JAK/STAT signaling of HSC-T6 cells].</a> Zhonghua Gan Zang Bing Za Zhi. 21 (9), 663-667 (2013).</li>
<li>Li, Y. K. [Some issues in methology of Chinese herbs serum pharmcology]. Zhong Yao Xin Yao Yu Lin Chuang Yao Li. 10 (5), 263(1999).</li>
<li>Zhang, L., et al. [A review of Chinese herbs serum pharmcology methodological study]. Nan Jing Zhong Yi Yao Da Xue Xue Bao. 18 (4), 254(2002).</li>
<li>Zhang, D. [Issues and strategies for study of serum pharmcology in oncology]. Zhong Yi Yan Jiu. 17 (5), 13-14 (2004).</li>
</ol>

Gli autori dichiarano di non avere interessi finanziari concorrenti.

Negli ultimi anni, più attenzione è stata prestata alle medicine a base di erbe, un tipo di medicina alternativa che è stato applicato nel contesto clinico nel mondo orientale per migliaia di anni. Diverso dal modello &quot;banco a letto&quot; della medicina moderna, la medicina tradizionale a base di erbe richiede innanzitutto il modello &quot;letto a panca&quot; per spiegare i propri meccanismi. Ciò può essere seguito dalla convalida eseguita in fase di panificazione e di elaborazione per lo sviluppo di nuovi far...

L&#39;interesse per lo studio e l&#39;applicazione della medicina tradizionale a base di erbe sta crescendo. Al contrario di farmaci moderni, in cui gli ingredienti chimici sono definiti, le formule a base di erbe hanno alcuni ingredienti sconosciuti e richiedono processi di estrazione per consentire la consegna dei loro composti attivi. Anche se molti studi cercano di selezionare un piccolo composto con una struttura ben nota come rappresentante dell&#39;intera erba o di formula a base di erbe, né le efficienze farmacologiche né i meccanismi possono essere considerati equivalenti 1 , 2 . Mentre l&#39;impronta digitale consente l&#39;analisi dei costituenti delle formule complesse a base di erbe, alcuni componenti non sono ancora chiaramente analizzati, portando ad una sfida quando si combinano tutti gli estratti per lo studio 3 . Le interazioni di numerosi costituenti / estratti mediano gli effetti terapeutici delle medicine a base di erbe. Per mantenere questo vantaggio, il tradizionale estratto forma-decottoIone-è ancora ampiamente utilizzato nella clinica. Poiché la procedura di estrazione ha un grande impatto sull&#39;efficacia terapeutica, è necessario un protocollo standard di decotto acqua tradizionale, specialmente per gli studi in vivo 4 , 5 . D&#39;altra parte, quando esplora i meccanismi farmacologici, è anche una sfida l&#39;amministrazione di decotti a base di erbe durante studi in vitro o ex vivo . Il concetto di un siero contenente droga fu proposto per la prima volta da Tashino nel 1988 6 . Da allora, un numero crescente di ricercatori lo hanno applicato alla medicina a base di erbe 7 , 8 , 9 . Anche se il metodo del siero contenente farmaci ha alcune limitazioni, come l&#39;influenza di determinate componenti del siero, è ancora considerato un metodo che imitava in modo preciso le condizioni fisiologiche. 10 , 11 , 12 , 13 . È stato inoltre applicato al trattamento dell&#39;anemia aplastica 14 , dell&#39;osteoporosi menopausa 15 , 16 , 17 , insufficienza ovarica precoce 18 , cancro mammario 19 , cancro ovarico 20 e pubertà ritardata 21 . Qui presentiamo protocolli dettagliati per la preparazione del decotto er-xian (EXD) e del suo siero contenente droga. Inoltre, descriviamo l&#39;applicazione del siero contenente EXD e EXD ai modelli osteoporotici della menopausa murina. Un EXD è composto da 9 g ciascuno di Curculigo orchioides Gaertn , Herbaa Epimedii </eM, Radix Morindae Officinalis e Radix Angelicae Sinensis e 6 g ciascuno di Cortex Phellodendri e Rhizoma Anemarrhenae per paziente adulto al giorno. La dose equivalente per un topo è 0.1418 EXD / kg / giorno sulla base della seguente equazione 22 : dB = dA * RB / RA * (WA / WB) 1/3 . DA e dB si riferiscono rispettivamente alla dose per peso corporeo (mg / kg) dell&#39;uomo e del mouse. La dose, in mg / kg, è sostituita dal numero di EXD / kg. RA e RB rappresentano rispettivamente il fattore del corpo umano e il fattore del corpo del topo proporzionale alla superficie del corpo (m 2 ) / peso corporeo (kg)) 2/3 (vedi tabella 1 ). WA e WB indicano rispettivamente il peso corporeo (kg) dell&#39;uomo e del mouse.

<table><tbody><tr><td>Curculigo orchioides Gaertn (9&amp;thinsp;g), Herbaa Epimedii (9&amp;thinsp;g), Radix Morindae Officinalis (9&amp;thinsp;g), Radix Angelicae Sinensis (9&amp;thinsp;g), Cortex Phellodendri (6&amp;thinsp;g), and Rhizoma Anemarrhenae (6&amp;thinsp;g)</td><td>Kang-qiao Chinese Medicine Yinpian Co. Ltd (Shanghai, CN)</td><td>160922</td><td>EXD components</td></tr><tr><td>Filter paper</td><td>GElifesciences</td><td>99-103-952</td><td>Filter EXD decoction before concentration</td></tr><tr><td>Imprinting Control Region (ICR) mice</td><td>Shanghai Laboratory Animal Center</td><td>SCXK 2007-0005</td><td>In vivo study</td></tr><tr><td>Sprague Dawley rats</td><td>Shanghai Laboratory Animal Center</td><td>SCXK 2007-0005</td><td>EXD-containing serum preparation</td></tr><tr><td>Syringe filter</td><td>Millipore</td><td>SLGP033RB</td><td>0.22 &amp;micro;m</td></tr><tr><td>gavage needles (10 ml)</td><td>Shanghai BO Ge trade sales department</td><td>59104274</td><td>Adminstration of EXD</td></tr><tr><td>Ketamine (80 mg/kg)&amp;nbsp;</td><td>Fujian Gutian Pharma Co. Ltd</td><td>H35020148</td><td>Anesthesia</td></tr><tr><td>Xylazine (10 mg/kg)</td><td>Sunway Pharma Co. Ltd</td><td>CB07591</td><td>Anesthesia</td></tr><tr><td>Dulbecco&amp;rsquo;s modification of Eagle&amp;rsquo;s medium Dulbecco (DMEM) culture medium</td><td>Gibco</td><td>12800-116</td><td>DMEM with 2 mM L-glutamine and without ribonucleosides and ribonucleotides</td></tr><tr><td>Streptomycin</td><td>Sigma</td><td>1277</td><td>100 &amp;micro;g / ml</td></tr><tr><td>Penicillin</td><td>Sigma</td><td>4687</td><td>100 &amp;micro;g / ml</td></tr></tbody></table>

preparation of herbal medicine: er-xian decoction and er-xian-containing serum for in vivo and in vitro experiments

La medicina tradizionale a base di erbe, una medicina alternativa nell&#39;ambiente clinico, ha ricevuto un&#39;attenzione maggiore negli ultimi anni. Prima della consegna al corpo, è necessaria una procedura di estrazione aggiuntiva per liberare i costituenti attivi dalle erbe grezze. La decozione dell&#39;acqua è una procedura di estrazione classica che è ancora ampiamente utilizzata nelle impostazioni cliniche. Qui proponiamo un protocollo dettagliato per la decotto er-xian (EXD) per applicare decotti a base di erbe a studi sperimentali. Viene descritto il calcolo di una dose appropriata per l&#39;animale, nonché le quattro fasi principali di EXD: l&#39;ammollo, la decozione dell&#39;acqua, la filtrazione e la concentrazione. Inoltre, EXD contenente siero viene introdotto nei ratti come mezzo di convalida in vitro . Qui, i ratti sono stati somministrati per via orale con EXD per tre giorni. I campioni di sangue sono stati raccolti, inattivati, centrifugati e filtrati. Il siero, diluito con il mezzo di coltura, può essere utilizzato per trattare cellule o tessuti in vItro. Ad esempio, EXD è stato applicato sia agli studi in vivo che in vitro e ha dimostrato che EXD migliora l&#39;osteogenesi. Questo protocollo può essere utilizzato come riferimento per la preparazione e l&#39;applicazione di medicinali a base di erbe.

L&#39;effetto di EXD sulla densità ossea dei topi OVX La colorazione ematossilina e eosinica (H &amp; E) della sezione vertebra lombare mostra una maggiore trabecola ossea dopo il trattamento EXD in vivo ( Figura 1A , pannello destro) rispetto a quelli del gruppo OVX ( Figura 1A , pannello a sinistra). Figura 1B mostra le immagini μCT rappresentative del 4 °</sup...

Watch this Scientific Journal Video about Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments at JoVE.com

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

Qui presentiamo la preparazione di un decotto er-xiano (EXD) in quattro fasi - immersione, decotto, filtrazione e concentrazione - e dimostrare la somministrazione di un siero preparato con EXD ai ratti. Questi metodi sono applicabili allo studio in vivo e in vitro di decotti a base di erbe come i medicinali cinesi tradizionali.

Preparazione della medicina a base di erbe: Eruzione di Er-Xian e Serum contenente Er-Xian per<em&gt; In Vivo</em&gt; E<em&gt; In vitro</em&gt; Esperimenti

Traditional herbal medicine, an alternative medicine in the clinical setting, has received increased attention in recent years. Before delivery to the ...

preparation-herbal-medicine-er-xian-decoction-er-xian-containing

Research

JoVE Journal

Medicine

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

10.9K Views.  Shanghai University of Traditional Chinese Medicine.  Qui presentiamo la preparazione di un decotto er-xiano (EXD) in quattro fasi - immersione, decotto, filtrazione e concentrazione - e dimostrare la somministrazione di un siero preparato con EXD ai ratti. Questi metodi sono applicabili allo studio in vivo e in vitro di decotti a base di erbe come i medicinali cinesi tradizionali. 

Video: Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

Extraction and Purification of Polyphenols from Freeze-dried Berry Powder for the Treatment of Vascular Smooth Muscle Cells In Vitro

Epidemiological studies indicate that increased flavonoid intake correlates with decreased mortality due to cardiovascular diseases (CVD) in the United States (US) and Europe. Berries are widely consumed in the US and have a high polyphenolic content. Polyphenols have been shown to interact with many molecular targets and to exert numerous positive biological functions, including antioxidant, anti-inflammatory, and cardioprotective effects. Polyphenols isolated from blackberry (BL), raspberry (RB), and black raspberry (BRB) reduce oxidative stress and cellular senescence in response to angiotensin II (Ang II). This work provides a detailed description of the protocol used to prepare the polyphenol extracts from freeze-dried berries. Polyphenol extractions from freeze-dried berry powder were performed using 80% aqueous ethanol and an ultrasonic-assisted extraction method. The crude extract was further purified and fractionated using chloroform and ethyl acetate, respectively. The effects of both crude and purified extracts were tested on Vascular Smooth Muscle Cells (VSMCs) in culture.

Extraction and Purification of Polyphenols from Freeze-dried Berry Powder for the Treatment of Vascular Smooth Muscle Cells In Vitro

This work details a step-by-step method to prepare polyphenol-rich extracts from freeze-dried berry powder. In addition, it provides a thorough description of how to use these polyphenol-rich extracts in cell culture in the presence of the peptide hormone angiotensin II (Ang II) using Vascular Smooth Muscle Cells (VSMCs).

Estrazione e purificazione di polifenoli da polvere di bacche congelata per il trattamento di cellule muscolari lisce vascolari<em&gt; In vitro</em

Epidemiological studies indicate that increased flavonoid intake correlates with decreased mortality due to cardiovascular diseases (CVD) in the United ...

Ricerca

Chimica

Ultra High-Performance Liquid Chromatography-Mass Spectrometry and Self-established Database Analysis of Chinese Herbal Medicine Components

Chinese herbal medicine is complex and has numerous unknown compounds, making qualitative research crucial. Ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) is the most widely used method in qualitative analysis of compounds. The method includes standardized and programmed protocols for sample pretreatment, MS tune, MS acquisition, and data processing. The sample pretreatments include collection, pulverization, solvent extraction, ultrasound, centrifugation, and filtration. Data post-processing was described in detail and includes data importing, self-established database construction, method establishment, data processing, and other manual operations. The above-ground part of the alpine yarrow herb, Achillea millefolium L., is used to treat inflammation, gastrointestinal disturbances, and pain and its 3-oxa-guaianolides could be useful leads for anti-inflammatory drug development. Three representative compounds in AML were identified, combining TOF-MS with a self-established database. Moreover, the differences from existing literature, liquid-phase parameter optimization, scan mode selection, ion source suitability, collision energy adjustment, isomer screening, method limitation, and possible solutions were discussed. This standardized analysis method is universal and can be applied to identify complex compounds in Chinese herbal medicine.

<meta charset="utf8"></head><body>Cromatografia liquida ad altissime prestazioni-spettrometria di massa e analisi del database auto-consolidato dei componenti della fitoterapia cinese

We describe a general protocol and systematic design that could be applied to separate and recognize complex components in alpine yarrow herb, Achillea millefolium L., a Chinese herbal medicine.

Cromatografia liquida ad altissime prestazioni-spettrometria di massa e analisi del database auto-consolidato dei componenti della fitoterapia cinese

Chinese herbal medicine is complex and has numerous unknown compounds, making qualitative research crucial. Ultra-high-performance liquid chromatography ...

Preparation Of Gushukang (GSK) Granules for In Vivo and In Vitro Experiments

Traditional Chinese herbal medicine plays a role as an alternative method in treating many diseases, such as postmenopausal osteoporosis (POP). Gushukang (GSK) granules, a marketed prescription in China, have bone-protective effects in treating POP. Before administration to the body, one standard preparation procedure is commonly required, which aims to promote the release of active constituents from raw herbs and enhance the pharmacological effects as well as therapeutic outcomes. This study proposes a detailed protocol for using GSK granules in in vivo and in vitro experimental assays. The authors first provide a detailed protocol to calculate the animal-appropriate dosages of granules for in vivo investigation: weighing, dissolving, storage, and administration. Second, this article describes protocols for micro-CT scanning and the measurement of bone parameters. Sample preparation, protocols for running the micro-CT machine and quantification of bone parameters were evaluated. Third, serum-containing GSK granules are prepared, and drug-containing serum is extracted for in vitro osteoclastogenesis and osteoblastogenesis. GSK granules were intragastrically administered twice per day to rats for three consecutive days. Blood was then collected, centrifuged, inactivated, and filtered. Finally, serum was diluted and used for performing osteoclastogenesis and osteoblastogenesis. The protocol described here can be considered a reference for pharmacological investigations of herbal prescription medicines, such as granules.

Preparation Of Gushukang GSK Granules for In Vivo and In Vitro Experiments

This article provides a detailed protocol for preparing a working solution of Gushukang granules for animal studies and GSK granule containing serum for in vitro experiments. This protocol can be applied to pharmacological investigations of herbal medicines as well as prescriptions for both in vivo and in vitro experiments.

Preparazione di Gushukang (GSK) Granuli per in vivo e in vitro

Traditional Chinese herbal medicine plays a role as an alternative method in treating many diseases, such as postmenopausal osteoporosis (POP). Gushukang ...

Medicina

Preparation of Naringenin Solution for In Vivo Application

The preparation of a compound (phytochemical) solution is an overlooked but critical step prior to its application in studies such as drug screening. The complete solubilization of the compound is necessary for its safe use and relatively stable results. Here, a protocol for preparing naringenin solution and its intraperitoneal administration in a high-fat diet and streptozotocin (STZ)-induced diabetic model is demonstrated as an example. A small amount of naringenin (3.52-6.69 mg) was used to test its solubilization in solvents, including ethanol, dimethylsulfoxide (DMSO), and DMSO plus Tween 80 reconstituted in physiological saline (PS), respectively. Complete solubilization of the compound is determined by observing the color of the solution, the presence of precipitates after centrifugation (2000 x g for 30 s), or allowing the solution to stand for 2 h at room temperature (RT). After obtaining a stable compound/phytochemical solution, the final concentration/amount of the compound required for in vivo studies can be prepared in a solvent-only (no PS) stock solution, and then diluted/mixed with PS as desired. The antidiabetic osteoporotic effects of naringenin in mice (intraperitoneal administration at 20 mg/kg b.w., 2 mg/mL) were assessed by measuring blood glucose, bone mass (micro-CT), and bone resorption rate (TRAP staining and ELISA). Researchers looking for detailed organic/phytochemical solution preparations will benefit from this technique.

Preparation of Naringenin Solution for In Vivo Application

Here, the protocol presents the preparation of naringenin solution for in vivo intraperitoneal administration. Naringenin is fully dissolved in a mixture of dimethylsulfoxide, Tween 80, and saline. The antidiabetic osteoporotic effects of naringenin were assessed by blood glucose testing, tartrate-resistant acid phosphatase staining, and enzyme-linked immunosorbent assay.

Preparazione della soluzione di Naringenin per l'applicazione in vivo

The preparation of a compound (phytochemical) solution is an overlooked but critical step prior to its application in studies such as drug screening. The ...

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules

The composition of the Tibetan medicine Rhodiola granules (RG) is complex, and the overall quality of RG is difficult to determine. Therefore, establishing a method to determine the multi-component in vitro dissolution of RG is of great significance for quality control. This study uses the second paddle method of the fourth general rule 0931 from the Chinese Pharmacopoeia (2020 edition), compliant with apparatus 2 of the United States Pharmacopeia (USP). The dissolution apparatus was set to a rotation speed of 100 rpm with ultrapure water as the dissolution medium. A sample volume of 1 mL was collected at each timepoint. Furthermore, the cumulative dissolution of gallic acid, salidroside, and ethyl gallic acid in RG at different time points was determined by high-performance liquid chromatography (HPLC). Finally, the dissolution curves were drawn, and the curves were fitted to the GompertzMod, the Gompertz, the Logistic, and the Weibull equations. The results showed that the cumulative dissolution of gallic acid in RG was over 80% at 1 min, the cumulative dissolution of salidroside and ethyl gallic acid was over 65% at 5 min, and the cumulative dissolution of each index component decreased after 30 min. The curve fitting demonstrated that the GompertzMod equation was the best-fitting model for each index component of RG. In conclusion, the dissolution test method described in this protocol is simple, accurate, and reliable. It can characterize the dissolution behavior of the index components in RG in vitro, which provides a methodological reference for quality control of RG and quality evaluation of other ethnic compounds.

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules

Here, we test the dissolution of Rhodiola granules (RG) in vitro, draw dissolution curves of salidroside, gallic acid, and ethyl gallate in ultrapure water, and fit the curves to different mathematical models. This protocol provides information and guidance for in vivo bioequivalence and in vivo-in vitro correlation studies of RG.

Una determinazione della dissoluzione in vitro di componenti multi-indice nella medicina tibetana Rhodiola granuli

The composition of the Tibetan medicine Rhodiola granules (RG) is complex, and the overall quality of RG is difficult to determine. Therefore, ...

Exploring the Two Herb Combination Strategy to Treat Injured PC12 Cells

In view of the advantages of the combination of traditional Chinese medicine (TCM) in the treatment of cerebral ischemia, we studied the differences in the efficacy and mechanism between the preparation combination and the component combination in order to explore the two herb combination strategy to treat injured PC12 cells. Cobalt chloride (CoCl2) combined with a glucose-free medium was employed to induce oxidative damage of PC12 cells. Then, the optimal combination of Astragalus mongholicus (Ast) and Erigeron breviscapus (Eri) injection was selected and combined following uniform design methods after screening their safe and effective concentration on PC12 cells. Further, the component combination screened comprises 10 &#181;M astragaloside A, 40 &#181;M scutellarin, and 75 &#181;M chlorogenic acid in two herbs. Then, MTT, Annexin V-FITC/PI, immunofluorescence, and Western blot analysis were used to evaluate the efficacy and mechanism of the preparation combination and the component combination on injured PC12 cells. The results showed that the optimal preparation combination for cell pro-survival was Ast injection and Eri capsule with a concentration of 6:1.8 (&#181;M). The component combination (10 &#181;M astragaloside A, 40 &#181;M scutellarin, and 75 &#181;M chlorogenic acid) was more effective than the preparation combination. Both combinations remarkably reduced apoptotic rate, the fluorescence intensity of caspase-3, and intracellular reactive oxygen species (ROS) level; meanwhile, they upregulated the expression levels of p-Akt/Akt, Bcl-2/Bax, and Nrf2. These effects were more evident in the component combination. In conclusion, both combinations can inhibit the injury induced by CoCl2 combined with a glucose-free medium on PC12 cells, thus promoting cell survival. However, the efficiency of the component combination over the preparation combination may be due to its stronger regulation of the PI3K/Akt/Nrf2 signaling pathway related to oxidative stress and apoptosis.

This protocol provides a combination strategy of two herbs to treat injured PC12 cells. The protocol provides a reference for optimizing the best application mode of traditional Chinese medicine (TCM).

Esplorare la strategia di combinazione di due erbe per trattare le cellule PC12 ferite

In view of the advantages of the combination of traditional Chinese medicine (TCM) in the treatment of cerebral ischemia, we studied the differences in ...

Optimization of the Epimedii Folium Mutton-Oil Processing Technology and Testing Its Effect on Zebrafish Embryonic Development

As a traditional Chinese medicine (TCM), Epimedii folium (EF) has a history in medicine and food that is &#62; 2,000 years old. Clinically, EF processed with mutton oil is often used as a medicine. In recent years, reports of safety risks and adverse reactions of products that use EF as a raw material have gradually increased. Processing can effectively improve the safety of TCM. According to TCM theory, mutton-oil processing can reduce the toxicity of EF and enhance its tonifying effect on the kidneys. However, there is a lack of systematic research and evaluation of EF mutton-oil processing technology. In this study, we used the Box-Behnken experimental design-response surface methodology to optimize the key parameters of the processing technology by assessing the contents of multiple components. The results showed that the optimal mutton-oil processing technology of EF was as follows: heating the mutton oil at 120 &#176;C &#177; 10 &#176;C, adding the crude EF, stir-frying it gently to 189 &#176;C &#177; 10 &#176;C until it is evenly shiny, and then removing it and cool. For every 100 kg of EF, 15 kg of mutton oil should be used. The toxicities and teratogenicities of an aqueous extract of crude and mutton-oil processed EF were compared in a zebrafish embryo developmental model. The results showed that the crude herb group was more likely to cause zebrafish deformities, and its half-maximal lethal EF concentration was lower. In conclusion, the optimized mutton-oil processing technology was stable and reliable, with good repeatability. At a certain dose, the aqueous extract of EF was toxic to the development of zebrafish embryos, and the toxicity was stronger for the crude drug than for the processed drug. The results showed that mutton-oil processing reduced the toxicity of crude EF. These findings can be used to improve the quality, uniformity, and clinical safety of mutton oil-processed EF.

In this protocol, the mutton-oil processing technology of Epimedii folium (EF) was optimized by applying a Box-Behnken experimental design-response surface methodology, and the effect of crude and optimized water-extracted EF on zebrafish embryonic development was preliminarily investigated.

Ottimizzazione della tecnologia di lavorazione dell'olio di montone Epimedii folium e test del suo effetto sullo sviluppo embrionale del pesce zebra

As a traditional Chinese medicine (TCM), Epimedii folium (EF) has a history in medicine and food that is &#62; 2,000 years old. Clinically, EF processed ...

<meta charset="utf8"></head><body>Impatto della formula Sanleng Jiashen sul carcinoma epatocellulare ortotopico nei topi

Establishment of Hepatocarcinoma in BALB/c-nu Mice and Investigation of the Therapeutic Effect of the Sanleng Jiashen Formula

The lethality of liver cancer and the resistance to chemical drugs have forced the search for effective prescriptions of traditional herbs for liver cancer. Animal models that are repeatable, easy to manipulate, and highly mimic the pathophysiological processes of liver cancer are the prerequisite for the successful screening of effective drug candidates. Meanwhile, reliable drug efficacy evaluation indicators and means are also the guarantee of anti-liver cancer drug research and development.
Sanleng Jiashen formula, a representative prescription of traditional Chinese medicine containing Sparganium stoloni erum, Buch. -Ham. (Sanleng), Panax ginseng C. A. Mey. (ginseng), Rheum officinale Baill. (rhubarb), and Ligusticum chuanxiong Hort. (Chuanxiong), is prescribed to nourish the liver and clear heat, remove toxins, and promote blood circulation to treat liver cancer. This experimental protocol describes the preparation of lyophilized Sanleng Jiashen formula and the establishment process of in-situ liver cancer in BALB/c-nu mice. Histopathological staining, immunohistochemical detection of cancer markers, in vivo imaging of mice, and chick embryo chorioallantoic membrane test were used to explore the inhibition and anti-angiogenesis effect of Sanleng Jiashen formula on malignant proliferation of liver cancer tissue. The data show that the Sanleng Jiashen formula can effectively resist the malignant proliferation of liver cancer tissue, which is manifested by reduced tumor mass volume, improved pathological damage, and lower levels of the cancer marker ki67.
The superior inhibition of angiogenesis also suggests that the Sanleng Jiashen formula may have the potential to treat and prevent the progression and deterioration of liver cancer. The whole experimental scheme shows a comprehensive process of traditional Chinese medicine components in the treatment of mouse liver cancer, which provides a reference for the establishment and optimization of a liver cancer model, as well as the research and development of drugs to prevent and treat liver cancer.

<meta charset="utf8"></head><body>Istituzione dell'epatocarcinoma nei topi BALB/c-nu e studio dell'effetto terapeutico della formula Sanleng Jiashen

Here, we present the step-by-step protocol to investigate the therapeutic and vascular inhibitory effects of the Sanleng Jiashen formula on BALB/c-nu mice with orthotopic hepatocellular carcinoma.

Istituzione dell'epatocarcinoma nei topi BALB/c-nu e studio dell'effetto terapeutico della formula Sanleng Jiashen

The lethality of liver cancer and the resistance to chemical drugs have forced the search for effective prescriptions of traditional herbs for liver ...

Preparazione della medicina a base di erbe: Eruzione di Er-Xian e Serum contenente Er-Xian per

Riepilogo

Esplora Altri Video

Estrazione e purificazione di polifenoli da polvere di bacche congelata per il trattamento di cellule muscolari lisce vascolari

Cromatografia liquida ad altissime prestazioni-spettrometria di massa e analisi del database auto-consolidato dei componenti della fitoterapia cinese

Cromatografia liquida ad altissime prestazioni-spettrometria di massa e analisi del database auto-consolidato dei componenti della fitoterapia cinese

Preparazione di Gushukang (GSK) Granuli per in vivo e in vitro

Preparazione della soluzione di Naringenin per l'applicazione in vivo

Una determinazione della dissoluzione in vitro di componenti multi-indice nella medicina tibetana Rhodiola granuli

Esplorare la strategia di combinazione di due erbe per trattare le cellule PC12 ferite

Ottimizzazione della tecnologia di lavorazione dell'olio di montone Epimedii folium e test del suo effetto sullo sviluppo embrionale del pesce zebra

Istituzione dell'epatocarcinoma nei topi BALB/c-nu e studio dell'effetto terapeutico della formula Sanleng Jiashen

Istituzione dell'epatocarcinoma nei topi BALB/c-nu e studio dell'effetto terapeutico della formula Sanleng Jiashen