Two new selariscinins from Selaginella tamariscina (Beauv.) Spring.

Two new selariscinins named selariscinin F (1) and selariscinin G (2), along with one known selariscinin D (3) were isolated from Selaginella tamariscina. The structures of 1-3 were elucidated on the basis of chemical and spectral analysis, including 1D, 2D NMR analyses and HRESIMS.

Characterization of estrogenic active ingredients in Cuscuta chinensis Lam. based on spectral characteristics and high­performance liquid chromatography/quadrupole time­of­flight mass spectrometry.

High-performance liquid chromatography (HPLC) is an efficient method that is widely used to assess the quality of traditional Chinese medicine (TCM). It is well known that the quality of TCM has a direct effect on its efficacy; therefore, in order to thoroughly explain how TCM exerts its efficacy, it is necessary to characterize its active ingredients and assess their quality. The application of the spectrum­effect method is crucial for determining the pharmacological basis of materials. The aim of the present study was to examine the correlation between chemical spectra and estrogenic activity of Cuscuta chinensis Lam., in order to reveal active compounds with potential therapeutic effects. The spectra of Cuscuta chinensis Lam. were recorded using HPLC, and estrogenic activity was determined using a uterus growth test and MTT assay. Combination of the results of bivariate analysis, principal component analysis and Gray relational analysis identified 19 active compounds, as follows: Quercetin­3­O­(2'­O­α­rhamnosy­6'­O­malony)­â€‹ß­D­glucoside, ka-empferol­3­O­ß­D­aplosyl­(1→2)­â€‹[­α­â€‹L­rhamnosy­(1→6)]­ß-wD-glucoside, 6­O­(E)­P­coumaroyl)­ß­â€‹D­fructofuranosyl­(2→1)­α­D­glucopyranoside, kaempferol­7­rhamnosy, kaempferol­3­ß­D-glucuronide, apigenin, 4­caffeoyl­5­coumaroylquinic acid, kaempferol­3­arabofuranoside, quercetin­3­O-ß­D-apiofuranosyl-(1→2)-ß­D­galactoside, dicaffeoylquinic acid, hyperin, quercitin, isorhamnetin, chlorogenic acid, quercetin, quercltrin­2''­gallate, quercetin­3, 7­α­L­dirhamnoside and stigmasterol, as well as one unknown compound. The present study laid a foundation for in vivo metabolic studies regarding Cuscuta chinensis Lam. and for the development of its clinical application.

HPLC/Q-TOF-MS-Based Identification of Absorbed Constituents and Their Metabolites in Rat Serum and Urine after Oral Administration of Cistanche deserticola Extract.

As a famous health food in China, Cistanche deserticola (C. deserticola) suggested an estrogenic activity according to our previous study. However, no one clarifies its active material basis to date. To find more potentially active constituents and elucidate metabolic pathways of metabolites, a method to simultaneously analyze multiple absorbed constituents and metabolites from C. deserticola in rat serum and urine was established using high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC/Q-TOF-MS). Based on HPLC/Q-TOF-MS method, a total of 24 components, involving 9 prototype constituents and 15 metabolites in rat serum and urine samples, were tentatively identified based on retention time, ultraviolet spectrum, MS data, compound fragmentation laws, published literatures, and reference substances. Most of the compounds existed in the form of metabolites. The proposed metabolic pathways of main metabolites were discussed, including methylation, demethylation, hydrolysis, hydroxylation, acetoxylation, glucuronidation, dehydrogenation, sulfation, esterification, and so on. Phenylethanoid glycosides were extensively metabolized and mutually transformed in vivo. This investigation provided valuable information for further study of the active ingredients and action mechanism of C. deserticola.

[Metabolism of recombinant CHO-GS cell reducing of toxic effect of ammonia].

The toxic effect of ammonia on rCHO-GS cell decreased obviously due to the transfection of GS system in serum-free culture. The maximum cell density, 15.6 x 10(5) cells/mL was obtained in the culture with 1.42 mmol/L ammonia. The growth of rCHO-GS cell was inhibited with an increased ammonia concentration. However, a cell density of 8.9 x 10(5) cells/mL was obtained when the concentration of ammonia was 12.65mmol/L. The intracellar metabolic pathways were affected due to the decrease of the toxic effect of ammonia on rCHO-GS cell. With the increase of initial ammonia concentration from 0.36mmol/L to 12.65mmol/L, the yield coefficients of cell to glucose and lactate to glucose decreased. The activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) increased by 43%, 140% and 25%, respectively, indicating that the utilization of glucose increased and the glycolysis pathway was more prone to efficient energy metabolism pathway. An increased activity of glutamate-pyruvate aminotransferase (GPT) showed that the conversation from glutamate to alpha-ketoglutarate was shifted to glutamate-pyruvate transamination pathway. The deamination pathway was inhibited due to a decreased activity of glutamate dehydrogenase. In addition, the number of cells in G0/G1 phase increased and the specific production rate of recombinant protein increased by 2.1-fold with the increase of initial ammonia concentration from 0.36mmol/L to 12.65mmol/L.

Estimation of Chinese hamster ovary cell density in packed-bed bioreactor by lactate production rate.

A method is described for estimating recombinant Chinese hamster ovary (rCHO) cell density in a packed-bed bioreactor by lactate production rate. The lactate production rate, which depended on both the cell numbers and cell growth rate, was modeled by segregating the cell population into two parts: one growing at a maximum specific growth rate and another non-growing. The individual cell in each part had the same lactate production rate. The established rate equation of lactate production matched the experimental data reasonably well and could be used to estimate the cell growth in the batch culture with microcarriers. Furthermore, in the perfusion culture of rCHO cells in a packed-bed bioreactor, the final cell density, 1.3 x 10(10) cells l(-1), estimated by lactate production rate, was comparable to the direct sample counting of 1.2 x 10(10) cells l(-1), showing that lactate production rate method would be useful in tracing the cell growth in packed-bed bioreactors.