Cinobufacini is an aqueous extract of Bufo bufo gargarizans Cantor dried skin, which has been widely used for cancer therapy in China. So far, its active components are still not very clear. In previous reports, bufadienolides with low-concentration were usually studied because of their anticancer effects. However, the high polarity constituents in cinobufacini are less investigated. The present study found that more than 50% contents of cinobufacini were water-soluble peptides. Then, in vitro anticancer experiments were carried out, including human stomach cancer cell lines BGC823 and MCG803, human colon cancer cell lines DLD-1 and HT-29, and human pancreatic cancer cell line MIAPACA-2. The IC50 for these cell lines model were ranged from 25-123 microgmL(-1). The results indicated that these peptides showed similar activity with cinobufacini injection. As a conclusion, this study provides a new and further understanding of anticancer components in cinobufacini injection.

To obtain chemical constituent information of rat plasma after oral administration of Huang-Lian-Jie-Du Decoction (HLJDD), a LC-FT-ICR-MS method has been established, and both positive and negative ions scan modes were include in the analysis. By comparing their retention time, high resolution mass data of HLJDD extracts, blank plasma and dosed plasma, 38 constituents, including 22 prototype compounds and 16 metabolites, were detected in rat plasma after oral administration of HLJDD. In the 22 prototype compounds, 16 constituents were determined unambiguously by comparing with references. In the analysis of metabolites, phase II reactions like glucuronidation and sulfation were the major biotransformation pathways of HLJDD. M11 was observed as the only phase I metabolite in present experiment. The results will be beneficial for the further pharmacokinetics and pharmacological evaluations of HLJDD.

Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.

In order to clarify the metabolism pathways of scandoside methyl ester, the analysis of metabolites profiling in four kinds of liver microsomes was performed by using an ultra-performance liquid chromatography/ electrospray-tandem mass spectrometry (UPLC-ESI-MS). The data obtained from the 0 h-incubation and the 2 h-incubation were compared and analyzed. After incubation, 5 metabolites of scandoside methyl ester were found in rat, Beagles, rhesus monkey and human liver microsome. The results showed that scandoside methyl ester's major metabolic pathway in the liver microsomes is hydrolysis, oxidation and reduction reactions, and there are certain kinds differences between species. The study provides a research base for further research about iridoid compounds in vivo metabolic pathways.

Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.

To identify the active components in Bufo melanostictus Schneider and clarify the difference between fresh and dried Venenum Bufonis, a UPLC-Orbitrap MS method has been established. The separation was performed with gradient elution of acetonitrile and water (with 0.1% formic acid) as mobile phase. By comparing their retention time and high resolution mass data of Venenum Bufonis extracts, 39 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were bufadienolides. There were significant differences in the main composition between fresh and dried Venenum Bufonis. There are fewer bufadienolides in fresh toad venom.

OBJECTIVETo establish a method for determination of geniposide in Beagle dogs plasma by high performance liquid chromatography (HPLC), and study the pharmacokinetics and bioavailability of geniposide in Beagle dogs after oral administration Xingnaojing.

METHODTo determine the geniposide in Beagle dogs plasma by HPLC after oral administration or intravenous injection Xingnaojing, and the pharmacokinetic parameters were calculated by the software of Kinetica.

RESULTThe good linearity range of geniposide was 1.24 - 158.88 mg x L(-1). The main pharmacokinetic parameters after oral administration was as follows: Cmax (11.8 +/- 0.6) mg x L(-1), Tmax (52.0 +/- 4.5) min, AUC(1280.8 +/- 172.0) mg x min x L(-1), MRT(118.7 +/- 25.4) min, and these parameters after intravenous injection was follows: Cmax 107.4 +/- 6.3) mg x L(-1), AUC(7930.1 +/- 670.0) mg x min x L(-1), MRT(92.4 +/- 5.1) min. The bioavailability of geniposide in Beagle dogs after oral administration Xingnaojing was (6.46 +/- 0.87)%.

CONCLUSIONThe HPLC method had good applicability. The extract recovery, method recovery, intra-day precision and inter-day precision of the method were all met the requirements. The stability in conditions of room temperature and freeze-thaw cycle was good. The results indicated that the oral administration bioavailability of geniposide was in low degree.

Administration, Oral ; Animals ; Biological Availability ; Chromatography, High Pressure Liquid ; Dogs ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Iridoids ; administration & dosage ; pharmacokinetics

OBJECTIVETo study and establish the optimal technology for the preparation of evodiae juice.

METHODThe contents of evodiamine, rutaecarpine and evodin were simultaneously determined with HPLC, and each yield of the three compounds were chosen as the evaluating indicator. The orthogonal test coupled with the weighted coefficient method were adopted to acquire the optimal technology for the preparation of evodiae juice.

RESULTThe study showed that the optimal technology for the preparation of evodiae juice was as follows: decocted three times while the first time was with 12-fold of water socked 30 minutes and decocted 45 minutes, the second time was with 8-fold of water decocted 20 minutes and the third time was with 6-fold of water decocted 20 minutes.

CONCLUSIONThis method is simple and accurate. The optimal technology is suitable for industry manufacture of evodiae juice.

Drugs, Chinese Herbal ; analysis ; Evodia ; chemistry ; Technology, Pharmaceutical ; methods

Objective To explore the effect of acteoside (one of the ingredients of Total Glycosides Extracted from Rehmannia capsules) on treatment of proteinuria and protection of podocytes.Methods In this study,puromycin nephropathy rat model was successfully established.After detecting the degree of proteinuria,the expression of podocyte injury markers and the degree of podocyte foot process fusion were investigated by electron microscope.In addition,puromycin treated podocyte injury model was also successfully established in vitro.Podocyte viability,migration,cytoskeleton and injury marker were detected.Results In vivo study showed that acteoside could effectively reduce proteinuria (P ＜ 0.05),restore the expression of podocyte injury markers such as nephrin and synaptopodin (all P ＜ 0.05),and alleviate the degree of podocyte foot process fusion.In vitro study showed that acteoside could effectively restored podocyte viability (P ＜ 0.05),reduce abnormal migration ability (P ＜ 0.05),protecte cytoskeleton and restore the expression of podocyte injury marker nephrin (P ＜ 0.05).Conclusions This study confirms that acteoside can reduce the degree of proteinuria in puromycin nephropathy rat model in vivo and alleviate the degree of podocyte injury in vitro as well as enrich the molecular mechanism of Total Glycosides Extracted from Rehmannia capsules in treatment of proteinuria.

Objective:To evaluate the clinical efficacy and safety of fecal microbiota transplantation(FMT)for the treatment of chronic functional constipation in the elderly.Methods:A total of 33 elderly patients with chronic functional constipation were included and given three sessions of FMT.Changes in fecal characteristics, constipation, mood and quality of life in these patients were evaluated using the Bristol stool form scale(BSFS), the constipation assessment scale(CAS), patient assessment of constipation symptoms(PAC-SYM), the Zung self-rating anxiety scale(SAS), the Zung self-rating depression scale(SDS), and the patient assessment of constipation quality of life(PAC-QOL)before and 12 weeks after treatment.The clinical efficacy was based on comparison between pre-and post-treatment results for each patient.Results:Clear improvement was achieved in 33 patients 12 weeks after treatment, compared with before transplantation.Post-treatment scores of the constipation assessment scale and symptom self-assessment questionnaire for patients with constipation were(8.9±1.2)scores and(26.5±2.4)scores, respectively, significantly lower than pre-transplantation scores of(12.2±1.1)scores and(32.4±2.4)scores( t=15.034, 13.904, both P<0.001). Similarly, post-treatment scores were also lower than pre-transplantation levels for the self-rating anxiety scale[(50.4±8.4)scores vs.(57.5±9.0)scores, t=10.333, P<0.001], the self-rating depression scale[(50.6±8.4)% vs.(55.0±10.5)%, t=5.301, P<0.001], and self-assessment questionnaire for quality of life[(88.2±7.3)scores vs.(103.7±7.3)scores, t=23.300, P<0.001]. Conclusions:FMT can improve fecal characteristics and constipation symptoms, relieve anxiety and depression, improve the quality of life, and provide a new option for the treatment for elderly patients with chronic functional constipation.