Transcriptomics reveals the effect of ammonia nitrogen concentration on Pseudomonas stutzeri F2 assimilation and the analysis of amtB function.

The biological treatment of wastewater with high concentrations of ammonia nitrogen has become a hot research issue, but there are limited reports on the mechanism of ammonia nitrogen utilization by microorganisms. In this paper, a transcriptomic approach was used to investigate the differences in gene expression at 500.0 mg/L (Amo 500) and 100.0 mg/L (Amo 100) ammonium concentrations to reveal the mechanism of ammonia nitrogen removal from water by Pseudomonas stutzeri F2. The transcriptome data showed 1015 (459 up-regulated and 556 down-regulated) differentially expressed genes with functional gene annotation related to nitrogen source metabolism, glycolysis, tricarboxylic acid cycle, extracellular polysaccharide synthesis, energy conversion and transmembrane transport, revealing the metabolic process of ammonium nitrogen conversion to biological nitrogen in P. stutzeri F2 through assimilation. To verify the effect of ammonium transporter protein (AmtB) of cell membrane on assimilation, a P. stutzeri F2-ΔamtB mutant strain was obtained by constructing a knockout plasmid (pK18mobsacB-ΔamtB), and it was found that the growth characteristics and ammonium removal rate of the mutant strain were significantly reduced at high ammonium concentration. The carbon source components and dissolved oxygen conditions were optimized after analyzing the transcriptome data, and the ammonium removal rate was increased from 41.23% to 94.92% with 500.0 mg/L ammonium concentration. The study of P. stutzeri F2 transcript level reveals the mechanism of ammonia nitrogen influence on microbial assimilation process and improvement strategy, which provides a new strategy for the treatment of ammonia nitrogen wastewater.

[Temporal-spatial Distribution and Health Risk Assessment of Heavy Metals in the Surface Water of Ningbo].

A total of 120 surface water samples were collected from industrial and commercial districts of Ningbo, China in the wet and dry seasons. The concentrations of six heavy metals (Cd, Cr, Ni, Pb, Zn, and Fe) in the samples were measured, the temporal-spatial distribution characteristics of the six heavy metals were analyzed, and Pearson correlation coefficients of the six heavy metals were calculated. Combined with the temporal-spatial distribution characteristics and Pearson correlation coefficients of the six heavy metals, the main pollution sources of the two districts were analyzed, respectively. The risk of heavy metals in surface water to the exposed population was evaluated by calculating the health risk index and carcinogenic risk index. The results showed that the pollution characteristics of heavy metals in the surface water of Ningbo industrial district and commercial district differed greatly in different seasons. In the industrial district, the orders of the average concentration of heavy metals in the wet season and dry season were Fe>Zn>Ni>Pb>Cr>Cd and Fe>Zn>Cr>Ni>Pb>Cd, respectively. The concentrations of Cr, Cd, and Pb in the wet and dry seasons exceeded the class Ⅳ recommended values, following the degrees of Cr>Cd>>Pb and Pb>Cr=Cd, respectively. Sewage containing heavy metals was one of the major pollution sources. In the commercial district, the average concentrations of heavy metals in the wet season and dry season were in the order of Fe>Pb>Ni>Zn>Cd>Cr and Fe>Pb>Ni>Zn>Cr=Cd, respectively. The concentrations of Cd and Pb in the wet season exceeded the corresponding levels (class Ⅳ), and the degree followed Cd>Pb. Only Pb exceeded the standard in the dry season, with the exceeding standard rate of 60%. Road pollution containing heavy metals was the major pollution source, and heavy metals entered surface water mostly with surface runoff and precipitation. The carcinogenic risk posed by heavy metals in the surface water of the Ningbo industrial district and commercial district was very high, and the carcinogenic risk in the commercial district was much higher than that in the industrial district. The main carcinogen was Cr. Compared to the research results of the research group in 2015, the pollution degree of heavy metals has been greatly reduced. In the future, we still need to give adequate attention to the prevention and control of heavy metal pollution in surface water in Ningbo.

[Anti-depression mechanism of Zuojin Pills:based on UHPLC-TOF-MS, network pharmacology, and experimental verification].

This study aims to explore the anti-depression mechanism of Zuojin Pills based on the plasma constituents, network pharmacology, and experimental verification. UHPLC-TOF-MS was used for qualitative analysis of Zuojin Pills-containing serum. Targets of the plasma constituents and the disease were retrieved from PharmMapper and GeneCards. Then the protein-protein interaction(PPI) network was constructed and core targets were screened for GO term enrichment and KEGG pathway enrichment. Cytoscape 3.7.2 was employed construct the "compound-target-pathway" network and the targets and signaling pathways of Zuojin Pills against depression were predicted. CUMS-induced depression mouse model was established to verify the key targets. The results showed that a total of 21 constituents migrating to blood of Zuojin Pills were identified, which were mainly alkaloids. A total of 155 common targets of the constituents and the disease and 67 core targets were screened out. KEGG enrichment and PPI network analysis showed that Zuojin Pills may play a role in the treatment of depression through AMPK/SIRT1, NLRP3, insulin and other targets and pathways. Furthermore, the results of animal experiments showed that Zuojin Pills could significantly improve the depression behaviors of depression, reduce the levels of IL-1ß, IL-6 and TNF-α in hippocampus and serum, activate AMPK/SIRT1 signaling, and reduce the protein expression of NLRP3. In conclusion, Zuojin Pills may play a role in the treatment of depression by activating AMPK/SIRT1 signaling pathway, and inhibiting NLRP3 activation and neuroinflammation in the hippocampus of mice.

[Assessment of Heavy Metal Pollution and Human Health Risk of Surface Waters in the City of Ningbo, China].

In order to evaluate the present situation and the potential ecological risk of heavy metal pollution in the surface waters of eastern Chinese coastal cities, the city of Ningbo was used as an example. From surface waters in Ningbo, 255 water samples were collected. The concentrations of six heavy metals (Cd, Cr, Cu, Ni, Pb, and Zn) were measured for the samples. The potential ecological risk of these heavy metals was evaluated by calculating health risk and cancer risk indices. The results indicate that, in the surface waters of Ningbo, the average concentrations of the six heavy metals were in the increasing order of Zn > Pb > Cr > Ni > Cd > Cu. Three of the heavy metal concentrations exceeded the national environmental quality standards for surface water (GB 3838-2002, grade Ⅴ), and the order of exceeding the standard:Cd > Pb > Cr. Electroplating wastewater, metal waste produced, traffic pollutant, dyes, and coatings waste were the main pollutant sources. There were some potential health risks in over 70 percent of the surface waters in Ningbo, and the surface waters of the Zhenhai and Haishu districts had the highest potential health risk. There were three major potential health risk heavy metals, and their order of increasing risk was Cd > Cr > Pb. For adults and juveniles, the average carcinogenic risk indices were 17600 and 24800 times the critical values, respectively. Moreover, over 95 percent of the risks were attributed to Cr. The potential carcinogenic risk index of commercial areas with dense population was nearly 2 times that of the city average. The results indicated that the state of heavy metal pollution in the surface waters of Ningbo was very serious, thus the government should pay more attention to this issue.

Inhibitory Effects of Danshen components on CYP2C8 and CYP2J2.

The use of Chinese herbal medicines and natural products has become increasingly popular in both China and Western societies as an alternative medicine for the treatment of diseases or as a health supplement. Danshen, the dried root of Salvia miltiorrhiza (Fam.Labiatae), which is rich in phenolic acids and tanshinones, is a widely used herbal medicine for the treatment of cardio-cerebrovascular diseases. The goal of this study was to examine the inhibitory effects of fifteen components derived from Danshen on CYP2C8 and CYP2J2, which are expressed both in human liver and cardiovascular systems. Recombinant CYP2C8 and CYP2J2 were used, and the mechanism, kinetics, and type of inhibition were determined. Taxol 6-hydroxylation and astemizole O-desmethyastemizole were determined as probe activities for CYP2C8 and CYP2J2, respectively. Metabolites formations were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results demonstrated that salvianolic acid A was a competitive inhibitor of CYP2C8 (Ki = 2.5 µM) and mixed-type inhibitor of CYP2J2 (Ki = 7.44 µM). Salvianolic acid C had moderate noncompetitive and mixed-type inhibitions on CYP2C8 (Ki = 4.82 µM) and CYP2J2 (Ki = 5.75 µM), respectively. Tanshinone IIA was a moderate competitive inhibitor of CYP2C8 (Ki = 1.18 µM). Dihydrotanshinone I had moderate noncompetitive inhibition on CYP2J2 (Ki = 6.59 µM), but mechanism-based inhibition on CYP2C8 (KI = 0.43 µM, kinact = 0.097 min-1). Tanshinone I was a moderate competitive inhibitor of CYP2C8 (Ki = 4.20 µM). These findings suggested that Danshen preparations appear not likely to pose a significant risk of drug interactions mediated by CYP2C8 after oral administration; but their inhibitory effects on intestinal CYP2J2 mediated drug metabolism should not be neglected when they are given orally in combination with other drugs. Additionally, this study provided novel insights into the underling pharmacological mechanisms of Danshen components from the perspective of CYP2C8 and CYP2J2 inhibition.

[Comparison of Heavy Metal Contamination Characteristics in Surface Water in Different Functional Areas: A Case Study of Ningbo].

From industrial and commercial areas of Ningbo City, China, 85 surface water samples were collected. The concentrations of six heavy metals (Cd, Cr, Cu, Ni, Pb, and Zn) in the samples were measured, and the characteristics of the spatial distributions of those metals were analyzed. Through a combination of regional characteristics and Pearson correlation coefficients of the different heavy metals, the main pollution sources of the two areas were analyzed. The potential ecological risks of these heavy metals were evaluated by considering health risk and cancer risk indices. The results indicate that the heavy metal pollution of surface waters was serious in both the industrial and commercial areas. Furthermore, the differences between the two areas were also observed. In the industrial area, the average concentrations of the six heavy metals were, from highest to lowest, in the order: Zn >>Ni>Pb> Cr> Cu> Cd, and the main sources were industrial emissions. Four heavy metal concentrations exceeded the national environmental standard, which, from highest to lowest, were in the order: Cd> Pb> Cr >>Zn. In the commercial area, the average concentrations, from highest to lowest, were in the order: Cr> Pb >>Zn> Ni> Cd> Cu, and the main sources were road pollutants. Three heavy metal concentrations exceeded the national standard, and the order, from highest to lowest, was Cd> Pb>>Cr, with Cd and Pb having the most potential health risk. In both the industrial and commercial areas, there were some potential health risks and high carcinogenic risks. Cd, Cr, and Pb have the highest potential health risks, whereas Cr is the major potentially carcinogenic metal. The commercial area had 1.7 times the potential carcinogenic risk as that of the industrial area. The government should pay more attention to heavy metal pollution of surface waters in Ningbo City, China.

[Clinical Effect of Treating Refractory Pressure Ulcers by Jinchuang Ointment].

Objective To observe the clinical effect of Jinchuang Ointment (JO) for treatment of refractory pressure ulcers. Methods Totally 306 patients with phase II , IIl, IV refractory pressure ul- cers were randomly assigned to the traditional Chinese herbal medicine ointment group, the JO group, the MepilexAg group,102 cases in each group. Each ulcer was scored using pressure ulcer scale for healing (PUSH) designed by National Pressure Ulcer Advisory Panel (NPUAP) before treatment, at day 7, 14, and 21 after treatment, respectively. The healing rate of pressure ulcer was observed at day 21. Results There was significant difference in PUSH score of the 3 groups between before treatment and at day 7/14)21 after treatment (P <0. 01). PUSH score was better in the JC group, as compared with that in the other two groups (P <0. 05) at day 14 and 21 after treatment. There was no significant difference in PUSH score between the traditional Chinese herbal medicine ointment group and the MepilexAg group (P >0. 05). Conclusions JO had significant effect in treatment of patients with phase II , III, IV pressure ulcers. The rate of wound healing at day 14/21 was significantly higher than that of traditional Chinese herbal medicine ointment and MepilexAg.

[LC-MS/MS method for determination of tripterine in plasma： pharmacokinetic study in Beagles].

To establish a LC-MS/MS method for determination of tripterine in Beagle plasma and study its pharmacokinetics after oral administration of tripterygium tablet. Plasma samples were extracted with dichloromethane and separated on a Phenomenex Luna C8 (2.0 mm×50 mm, 3 µm) column with methanol-acetonitrile isopropanol(1∶1)-1‰formic acid (15∶55 ∶30) as the mobile phase. Tripterine ([M+H] ⁺, m/z 451.3/201.1) and internal standard prednisolone ([M+H] ⁺, m/z 361.1/147.1) were monitored in multiple reaction monitoring (MRM). The concentration-time curves were simulated by drug and statistic software 1.0 and the pharmacokinetic parameters were calculated. There was a good linear relationship between peak area ratio and concentration of tripterine and internal standard prednisolone within range of 0.680 0-136.0 µg•L⁻¹. The limit of quantitation was 0.680 0 µg•L⁻¹ and the intra- and inter-day precision was within 6.15%. The absolute recovery rate was between 50.42% to 51.65%. The concentration-time curves were consistent with the one-compartment model(w=1/cc). The main pharmacokinetic parameters after a single dose were as follows： Cmax (35.64±9.540) µg •L⁻¹,Tmax(2.62±0.69) h,T1/2(2.93±0.29) h, CL (0.308±0.056) L•kg⁻¹â€¢h⁻¹, AUC0-12 (131.16±31.94) µg•L•h⁻¹, AUC0-∞ (142.83±37.57) µg•L•h⁻¹. The established LC-MS/MS method was proved to be sensitive, accurate and convenient, suitable for the pharmacokinetic study of Tripterygium tablet in Beagle dogs.

Pharmacokinetic profiles of hydroxysafflor yellow A following intravenous administration of its pure preparations in healthy Chinese volunteers.

ETHNOPHARMACOLOGICAL RELEVANCE: Hydroxysafflor yellow A (HSYA), the major active marker compound isolated from Carthamus tinctorius L., has been demonstrated to possess various attractive pharmacological activities. However, there is a lack of information about the complete clinical pharmacokinetic profiles of HSYA following the administration of its pure preparations. The purpose of this study was to fully characterize the pharmacokinetic (PK) properties of HSYA in healthy Chinese volunteers following drip intravenous infusion of injectable powder of pure HSYA (IPPH), a new drug recently approved for the phase I clinical study by China Food and Drug Administration. MATERIALS AND METHODS: 36 healthy subjects of either sex were recruited in this single-center, and open-label, single doses (25, 50, and 75 mg) and multiple doses (50 mg, once daily, 7 consecutive days) study. Plasma samples were analyzed with a validated LC-MS/MS method. Various PK parameters were estimated from the plasma concentration versus time data using non-compartmental methods. RESULTS: After single dose administration of IPPH, the values of AUC(0-t), AUC(0-∞) and C(max) for HSYA were statistically proportional over the dose range of 25-75 mg. After 7 repeated doses of 50 mg IPPH, both C(max) and AUC(0-∞) were significantly decreased, from 3207 to 2959 µg L(-1), and from 12,811 to 12,135 µg h L(-1) respectively, while t(1/2) was significantly prolonged from 3.912 to 4.414 h. The minimum plasma concentrations on day 5, 6 and 7 showed good stability with no significant difference. Both Cmax and AUC of HSYA in male volunteers were generally lower than that in females. IPPH was generally well tolerated in healthy volunteers by either single or multiple dosing. CONCLUSION: HSYA displayed moderately linear PK properties over the doses ranging from 25 to 75 mg of IPPH. Repeated administration of IPPH once daily could not lead to the in-vivo drug accumulation, but significantly affect PK behavior of HSYA. Gender difference should be considered for dosage recommendation in the clinic.

A correlative study of polymorphisms of CYP2C19 and MDR1 C3435T with the pharmacokinetic profiles of lansoprazole and its main metabolites following single oral administration in healthy adult Chinese subjects.

Considering that the genotypes of CYP2C19 and MDRI C3435T are two major factors attributed to the inter-individual pharmacokinetic variability of lansoprazole (LSZ), the aim of the study was to simultaneously elucidate the effects of CYP2C19 and MDRI C3435T polymorphisms on the pharmacokinetics difference of LSZ and its metabolites 5'-hydroxy lansoprazole (HLSZ) and lansoprazole sulphone (LSZS) following oral administration of LSZ tablets in healthy Chinese subjects. Plasma concentration of LSZ, HLSZ and LSZS were quantified by a sensitive and specific LC-MS/MS method, while the genotypes of CYP2C19 and MDRI C3435T for each subject were identified by a direct sequencing method. Statistical analysis was performed in the pharmacokinetic parameters including Cmax, t1/2, Tmax, MRTo_-, AUCO-2 and AUCo_r among different genotype groups of CYP2C 19 and MDRI C3435T. Compared to the CYP2Cl9 EMs, the CYP2C 19 PM group showed slower elimination and betteroral bioavailability of LSZ, much higher plasma concentrations of LSZS and lower concentrations of HLSZ with statistically significance. Despite a tendency of more favorable absorption and rapid elimination of LSZ in wild genotype, no significant pharmacokinetics difference was observed between the wild genotype of MDR1 C3435T and its mutant types. In conclusion, the pharmacokinetics of MDRI C3435T.

Enhanced riboflavin production by recombinant Bacillus subtilis RF1 through the optimization of agitation speed.

Dissolved oxygen is one of the most important bioprocess parameters that could affect cell growth and product formation, and it is easy to control by changing agitation speed. In this work, the effects of agitation speed on the performance of riboflavin production by recombinant Bacillus subtilis RF1 was investigated in fed-batch fermentation. The lower agitation speed (600 rpm) was beneficial for cell growth and riboflavin biosynthesis in the initial phase of fermentation process. While, during the later phase, higher agitation speed (900 rpm) was favor for cell growth and riboflavin biosynthesis. Thus, a two-stage agitation speed control strategy was proposed based on kinetic analysis, in which the agitation speed was controlled at 600 rpm in the first 26 h and then switched to 900 rpm to maintain high µ for cell growth and high q(p) for riboflavin production during the entire fermentation process. However, it was observed that a sharp increase of agitation speed resulted in an adverse effect on cell growth and riboflavin synthesis within a short time. To avoid this phenomenon, a multi-stage agitation speed control strategy was set up based on the two-stage control strategy, the maximum concentration of riboflavin reached 9.4 g l(-1) in 48 h with the yield of 0.051 g g(-1) by applying this strategy, which were 20.5 and 21.4% over the best results controlled by constant agitation speeds.

Fermentation of biodiesel-derived glycerol by Bacillus amyloliquefaciens: effects of co-substrates on 2,3-butanediol production.

Cultivation in glycerol instead of sugars inhibits 2,3-butanediol (2,3-BD) production by Bacillus amyloliquefaciens. In this study, we report that B. amyloliquefaciens readily produces 2,3-BD from biodiesel-derived glycerol in the presence of beet molasses as a co-substrate. Unexpectedly, the molasses stimulated 2,3-BD production and simultaneously reduced the duration of fermentation. Productivity of 2,3-BD was enhanced at the start of fermentation, and yields increased under continuous molasses supply. Subsequently, 2,3-BD production in molasses-supplemented fed-batch culture was observed. Prior to inoculation of fed-batch fermentation culture, 15 g/l of molasses was added to the bioreactor. After 6 h of incubation, the bioreactor was fed with a solution containing 80 % glycerol and 15 % molasses. The 2,3-BD concentration, yield, and productivity significantly improved, reaching 83.3 g/l, 0.42 g/g, and 0.87 g/l·h, respectively. To our knowledge, these results are the highest report for 2,3-BD fermentation from biodiesel-derived glycerol.

Simultaneous characterization of prenylated flavonoids and isoflavonoids in Psoralea corylifolia L. by liquid chromatography with diode-array detection and quadrupole time-of-flight mass spectrometry.

RATIONALE: Prenylated flavonoids and isoflavonoids are widely distributed throughout the plant kingdom, with many biological effects. Psoralea corylifolia, which contains many kinds of prenylated components, has been widely used as a medicinal plant in Asia and India for thousands of years. The goal of this study was to characterize the components in P. corylifolia using a liquid chromatography with diode-array detection and quadrupole time-of-flight mass spectrometry (LC-DAD/Q-TOF-MS) method, and to elucidate the fragmentation behavior of the different prenyl substituent groups and their appropriate characteristic pathways in positive ion mode. METHODS: The calculated accurate masses of the protonated molecules, the fragment ions, the retention behavior, and the data from UV spectra were used for identification of the components in P. corylifolia. RESULTS: A total of 45 compounds, including 43 prenylated components, were identified or tentatively identified in P. corylifolia. Different diagnostic fragment ions and neutral losses were observed in different prenyl substructures: neutral loss of 56 Da (C(4)H(8)) and a fragment ion at m/z 69 (C(5)H(9)(+)) were generated by a prenyl chain; neutral losses of 42 Da (C(3)H(6)), 54 Da (C(4)H(6)), 15 Da (CH(3•)) and 16 Da (CH(4)) were observed in a ring-closed prenyl group; neutral losses of 72 Da (C(4)H(8)O), 60 Da (C(2)H(4)O(2)), 58 Da (C(3)H(6)O) and 18 Da (H(2)O) were detected in a 2,2-dimethyl-3,4-dihydroxydihydropyran ring; neutral losses of 72 Da (C(4)H(8)O), 60 Da (C(3)H(8)O) and 18 Da (H(2)O) were yielded from a 2,2-dimethyl-3-hydroxydihydropyran ring, a 2-(1-hydroxy-1-methylethyl)dihydrofuran ring or a 1-hydroxy-3-methylbut-3-enyl chain. CONCLUSIONS: This method can be applied for analysis of prenylated components in P. corylifolia and other herbal medicines.

Oral bioavailability and gender-related pharmacokinetics of celastrol following administration of pure celastrol and its related tablets in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Celastrol is a natural compound extracted from the traditional Chinese medicinal herb, Thunder God Vine (TGV). Owing to its potential anti-inflammatory and antitumor effects, celastrol has been considered as a promising candidate for drug development. AIM OF THE STUDY: To establish a sensitive LC-MS/MS method to investigate the pharmacokinetic properties of celastrol in rats. Key pharmacokinetic issues of celastrol including oral bioavailability, comparative pharmacokinetics between pure compound and tablet preparation, as well as gender-related pharmacokinetic difference are to be addressed for the first time. MATERIALS AND METHODS: Sprague-Dawley rats were administrated an intravenous dose (100 µg kg(-1)) of pure celastrol and an oral dose (1000 µg kg(-1)) of pure celastrol and TGV tablets (corresponding to 534 µg kg(-1) of celastrol), respectively. At different time points, the concentration of celastrol in rat plasma was determined by a sensitive and well-validated LC-MS/MS method. Main pharmacokinetic parameters including area under the plasma concentration-time curve (AUC), maximal plasma concentration (Cmax), the time for maximal concentration (Tmax) and mean residence time (MRT) were estimated by Drug and Statistic1.0 pharmacokinetic software (Chinese Pharmacological Association, Anhui, PR China). Statistical analysis was performed using two one-side t test with p-values less than 0.05 as the level of significance. RESULTS: The standard curve of celastrol showed good linearity in the concentration range of 0.11~54.3 ng mL(-1) in our current method, with acceptable selectivity, precision, recovery, and stability. The oral absolute bioavailability of celastrol significantly increased from 17.06% for pure celastrol to 94.19% for TGV tablets containing equivalent celastrol. After oral administration of TGV tablets, the Cmax and AUC values of celastrol in female rats were (32.03±8.41) µg L(-1) and (379.49±118.19) µg h L(-1), which were significantly higher (p<0.01) than that in males with the values of (14.31±7.33) µg L(-1) and (188.17±92.33) µg h L(-1). CONCLUSION: Celastrol administered orally in the rat was poorly absorbed into the systemic circulation. However, the poor absorption of celastrol could be greatly improved when celastrol-containing TGV tablets orally administered, and thereby the oral bioavailability of celastrol was significantly increased. As for gender difference, female rats showed significantly better absorption of celastrol than males.

[Efficacy of ginsenosides combined with prednisone in patients with systemic lupus erythematosus: a prospective, randomized, double-blind, placebo-controlled trial].

BACKGROUND: The side effects of glucocorticoid in treatment of systemic lupus erythematosus (SLE) have been the focus of debate, and our preliminary study indicates that ginsenosides can enhance the efficacy of dexamethasone. OBJECTIVE: To observe the effects of ginsenosides combined with prednisone in SLE patients. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: A total of 60 SLE patients from Department of Rheumatology and Immunology, Changhai Hospital, Second Military Medical University, were randomly divided into treatment group and control group, with 30 patients in each group. Patients in the treatment group were given routine treatment with prednisone plus ginsenosides, while those in the control group were given routine treatment with prednisone plus placebo. They were all treated for 3 months. MAIN OUTCOME MEASURES: After three-month treatment, syndrome score in traditional Chinese medicine (TCM), total response rate and symptom improvement rate were measured and evaluated. RESULTS: Twenty-eight cases in treatment group and twenty-seven cases in control group were included in analysis. The total response rates in the treatment group and control group were 89.28% and 66.67% respectively, and there was a significant difference between the two groups (P<0.05). After treatment, the TCM syndrome scores in the two groups were lower than those before treatment (P<0.01), and prednisone plus ginsenosides was better in decreasing the TCM syndrome score than prednisone plus placebo (P<0.05). The symptoms were improved in the treatment group as compared with the control group (P<0.05). CONCLUSION: Prednisone combined with ginsenosides can increase the clinical effective rate and improve the clinical symptoms of SLE patients.

Pharmacokinetic characterization of ginsenoside Rh2, an anticancer nutrient from ginseng, in rats and dogs.

The pharmacokinetic characteristics of ginsenoside Rh2, an anticancer nutrient, were analyzed in dogs and rats, including plasma kinetics, bioavailability, tissue distribution, plasma protein binding and excretion. The bioavailability of Rh2 is about 5% in rats and 16% in dogs. Multiple-dosing (7 days, 1 mg/kg bid) did not affect the pharmacokinetics in dogs. After oral dosing, Rh2 distributed mainly to the liver and gastrointestinal tissues in rats. In rats, the circulating fraction of Rh2 bound to plasma proteins was around 70%. The systemic clearance, however, was low -- around 2 and 20 ml/min/kg in dogs and rats, respectively. Only 1% of dosed Rh2 were recovered in excreta of rats as the intact form after oral administration, while 30% was excreted unchanged in bile after i.v. dosing. We subsequently investigated the membrane permeability of Rh2 across Caco-2 cell monolayers, stability and elimination profiles in the gastrointestinal environment. Low membrane permeability (P(app)(AP-BL): 1.91 x 10(-8)cm/s), efflux transport (efflux ratio: 9.8), pre-systemic elimination (degradation in acidic condition; metabolism in intestine tissue and contents), as well as low solubility largely accounted for the low bioavailability of Rh2. Regarding the low solubility of Rh2, micronization of the dose almost doubled the rate of absorption in dogs. Preliminary metabolite profiling confirmed the presence of the deglycosidating product protopanaxadiol in rat feces. A possible metabolite in rat bile and a potential sulfate-conjugate in rat urine were also detected.

Gender difference regarding schizandrin pharmacokinetics in rats.

The difference in the pharmacokinetics of schizandrin (SZ) in male and female rats was studied. SZ concentrations in the plasma were determined after the intragastric (i.g) administration of 10 mg/kg and the intravenous (i.v) administration of 5 mg/kg in male and female rats, respectively. It was found that the plasma concentrations of SZ in female rats were significantly higher than those in male rats. Drug absolute bioavailability, based on the area under curve (AUC(0-tn)), in female rats was roughly 20 times of that in male rats. The terminate half-life (T(1/2)) in male rats was shorter than that in female rats. These results demonstrate the existence of marked gender differences for SZ in rats.

[Enzyme kinetics of schizandrin metabolism and sex differences in rat liver microsomes].

To study the enzyme kinetics of schizandrin metabolism in different gender in rat liver microsomes, liver microsomes were prepared from male or female rats. Schizandrin was incubated with rat liver microsomes. Schizandrin and its metabolites were isolated and identified by HPLC-UV method. Vmax, Km and Cl(int) of schizandrin in male and female rat liver microsomes were (21.88 +/- 2.30) and (0.61 +/- 0.07) micromol x L(-1) x min(-1) x mg(-1) (protein), (389.00 +/- 46.26) and (72.64 +/- 13.61) micromol x L(-1), (0.0563 +/- 0.0007) and (0.0084 +/- 0.0008) min x mg(-1) (protein), respectively. The major metabolites of schizandrin in female and male rat liver microsomes were 7,8-dihydroxy-schizandrin (M1) and 7, 8-dihydroxy-2-demethyl schizandrin (M2b), respectively. Ketoconazole, quinidine, and orphenadrine had different level effects on schizandrin metabolism in both male and female rat liver microsomes, and cimetidine still had some inhibitory effect in male liver microsomes. CYP3A and CYP2C11 may be the main P450 enzymes in schizandrin metabolism and their difference in rat liver microsomes may be the main reason for the sex difference of metabolic enzyme kinetics and metabolites of schizandrin in rats.

Determination of ginsenoside Rd in dog plasma by liquid chromatography-mass spectrometry after solid-phase extraction and its application in dog pharmacokinetics studies.

A sensitive liquid chromatography-mass spectrometric (LC/MS) method for the quantification of ginsenoside Rd in dog plasma was developed and validated after solid-phase extraction (SPE). Chromatographic separation was achieved on a reversed-phase Cromosil C(18) column with the mobile phase of acetonitrile-ammonium chloride (500 micromol/L) and step gradient elution resulted in a total run time of about 5.5 min. The analytes were detected by using an electrospray negative ionization mass spectrometry in the selected ion monitoring (SIM) mode. A good linear relationship was obtained in the concentration range studied (0.005-2.500 microg/mL) (r=0.9998). Lower limit of quantification (LLOQ) was 5 ng/mL by using 500 microL plasma sample. Average recoveries ranged from 70.71 to 75.89% in plasma at the concentrations of 0.010, 0.100 and 2.500 microg/mL. Intra- and inter-day relative standard deviations were 8.49-11.71 and 5.71-16.48%, respectively. This method was successfully applied to the pharmacokinetic studies on dogs. The absolute bioavailability of Rd in dogs was 0.26%.

Development and validation of a sensitive quantification method for hematoporphyrin monomethyl ether in plasma using high-performance liquid chromatography with fluorescence detection.

A rapid, sensitive, precise and specific method for determination of hematoporphyrin monomethyl ether (HMME), a novel photodynamic therapy (PDT) drug, was developed and validated using high-performance liquid chromatography (HPLC) with fluorescence detection. HMME was isolated from the plasma by a single-step liquid-liquid extraction with ethyl acetate. The analyte and internal standard fluorescein were baseline separated on a Diamonsil C(18) analytical column (4.6 x 150 mm, 5 microm) and analyzed using a fluorescence detector with the excitation and emission wavelengths set at 395 and 613 nm, respectively. The method was linear in the concentration range 0.025-5 microg/mL with a lower limit of quantitation (LLOQ) of 10 ng/mL. The inter- and intra-day accuracies and precisions were all within 10% and the mean recoveries of HMME and fluorescein were 95 +/- 3.7 and 90 +/- 2.3%, respectively. The analyte was stable during all sample storage, preparation and analysis periods. This method was successfully applied to a pharmacokinetic study after a single-dose intravenous administration of HMME (5 mg/kg) to beagle dogs. This method was reproducible and sensitive enough for the pharmacokinetic study of HMME. Based on the results of the pharmacokinetic study, we suggest that a rather long light-avoiding time is essential for patients under HMME therapy.