A review on the pharmacokinetics of paeoniflorin and its anti-inflammatory and immunomodulatory effects.

Increasing pharmacological evidence supports that paeoniflorin, a water-soluble monoterpene glycoside isolated from Paeonia lactiflora Pall. (Shaoyao in Chinese), has a wide range of medicinal properties including anti-inflammatory, antioxidant, antithrombotic, anticonvulsive, analgesic, cardioprotective, neuroprotective, hepatoprotective, antidepressant-like, antitumoral, and immune-regulatory activities; as well as enhancing cognition and attenuating learning impairment. In addition to pharmacodynamic studies, information on pharmacokinetics is also significant for the further development and utilization of paeoniflorin. The present review focuses on the absorption, distribution, metabolism, and excretion of paeoniflorin, especially main pharmacological activities of paeoniflorin on inflammation and immune function. According to the findings obtained both in vitro and in vivo, a broad application prospect has been opened for paeoniflorin. However, further studies are needed to clarity the direct molecular mechanisms and key targets underlying the beneficial effects of paeoniflorin on inflammation and immunity.

Investigation of Efficacy Enhancing and Toxicity Reducing Mechanism of Combination of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba in Adjuvant-Induced Arthritis Rats by Metabolomics.

Combination of Aconiti Lateralis Radix Praeparata (FZ) and Paeoniae Radix Alba (BS) shows a significant effect in rheumatoid arthritis (RA). This study aimed to investigate the efficacy enhancing and toxicity reducing mechanism of combination of them in adjuvant-induced arthritis (AIA) rats by metabolomics. Rats were randomly divided into seven groups, including A (healthy control), B (model control), C1 (therapy group), C2 (efficacy enhancing group), D1 (toxicity group), and D2 (toxicity reducing group), and dexamethasone group was used as positive control. The plasma biochemical indexes showed that therapeutic dose of lipid-soluble alkaloids of FZ could significantly inhibit the concentrations of IL-1ß, TNF-α, and IFN-γ in AIA rats, and combination with total glucosides of peony could further reduce the concentration of IL-1ß. Then, UPLC-LTQ/Orbitrap MS with untargeted metabolomics was performed to identify the possible metabolites and pathways. Through multivariate data analysis of therapeutic dose groups (A vs. B vs. C1 vs. C2) and multivariate data analysis of toxic dose groups (A vs. B vs. D1 vs. D2), 10 and 7 biomarkers were identified based on biomarker analysis, respectively. After inducing AIA model, the plasma contents of spermidine, vanillylmandelic acid, catechol, and linoleate were increased significantly, and the contents of citric acid, L-tyrosine, L-phenylalanine, leucine, L-tryptophan, and uridine 5'-monophosphate (UMP) were decreased significantly. High dose of lipid-soluble alkaloids of FZ could increase the plasma contents of L-lysine, L-arginine, and deoxycholic acid, while the plasma contents of UMP, carnitine, N-formylanthranilic acid, and adenosine were decreased significantly. The pathway analysis indicated that therapeutic dose of lipid-soluble alkaloids of FZ could regulate energy and amino acid metabolic disorders in AIA rats. However, toxic dose could cause bile acid, fat, amino acid, and energy metabolic disorders. And combination with total glucosides of peony could enhance the therapeutic effects and attenuate the toxicity induced by lipid-soluble alkaloids of FZ.

MicroRNA-197 controls ADAM10 expression to mediate MeCP2's role in the differentiation of neuronal progenitors.

Duplication of MECP2 (Methyl-CpG-binding protein 2) causes severe mental illness called MECP2 duplication syndrome (MDS), yet the underlying mechanism remains elusive. Here we show, in Tg(MECP2) transgenic mouse brain or cultured neural progenitor cells (NPCs), that elevated MeCP2 expression promotes NPC differentiation into neurons. Ectopic expression of MeCP2 inhibits ADAM10 and thus the NOTCH pathway during NPC differentiation. In human cells, this downregulation on ADAM10 was mediated by miRNA-197, which is upregulated by MeCP2. Surprisingly, miR-197 binds to the ADAM10 3'-UTR via its 3' side, not the canonical seed sequence on the 5' side. In mouse cells, a noncoding RNA Gm28836 is used to replace the function of miR-197 between MeCP2 and ADAM10. Similar to MeCP2, overexpressing miR-197 also promotes NPCs differentiation into neurons. Interestingly, three rare missense mutations (H371R, E394K, and G428S) in MECP2, which we identified in a Han Chinese autism spectrum disorders (ASD) cohort showed loss-of-function effects in NPC differentiation assay. These mutations cannot upregulate miR-197. Overexpressing miR-197 together with these MeCP2 mutations could rescue the downregulation on ADAM10. Not only the inhibitor of miR-197 could reverse the effect of overexpressed MeCP2 on NPCs differentiation, but also overexpression of miR-197 could reverse the NPCs differentiation defects caused by MECP2 mutations. Our results revealed that a regulatory axis involving MeCP2, miR-197, ADAM10, and NOTCH signaling is critical for NPC differentiation, which is affected by both MeCP2 duplication and mutation.

Investigation of Liver Injury of Polygonum multiflorum Thunb. in Rats by Metabolomics and Traditional Approaches.

Liver injury induced by Polygonum multiflorum Thunb. (PM) have been reported since 2006, which aroused widespread concern. However, the toxicity mechanism of PM liver injury remained unclear. In this study, the mechanism of liver injury induced by different doses of PM after long-term administration was investigated in rats by metabolomics and traditional approaches. Rats were randomly divided into control group and PM groups. PM groups were oral administered PM of low (10 g/kg), medium (20 g/kg), high (40 g/kg) dose, while control group was administered distilled water. After 28 days of continuous administration, the serum biochemical indexes in the control and three PM groups were measured and the liver histopathology were analyzed. Also, UPLC-Q-TOF-MS with untargeted metabolomics was performed to identify the possible metabolites and pathway of liver injury caused by PM. Compared with the control group, the serum levels of ALT, AST, ALP, TG, and TBA in middle and high dose PM groups were significantly increased. And the serum contents of T-Bil, D-Bil, TC, TP were significantly decreased. However, there was no significant difference between the low dose group of PM and the control group except serum AST, TG, T-Bil, and D-Bil. Nine biomarkers were identified based on biomarkers analysis. And the pathway analysis indicated that fat metabolism, amino acid metabolism and bile acid metabolism were involved in PM liver injury. Based on the biomarker pathway analysis, PM changed the lipid metabolism, amino acid metabolism and bile acid metabolism and excretion in a dose-dependent manner which was related to the mechanism of liver injury.

[Effect of processing time of Polygoni Multiflori Radix on content changes of 16 componens].

To study 48 h processing time of Polygoni Multiflori Radix on its contents and changes of chemical components. HPLC was used to determine the contents of various components in 22 Polygoni Multiflori Radix samples with different processing time, and then the fingerprint similarity analysis and clustering analysis were used for characteristics analysis. Results showed that the similarity was between 0.9-1.0, with good correlation between the samples. In the clustering analysis, the 22 Polygoni Multiflori Radix and processed Polygoni Multiflori Radix samples were classified into 4 types according to the composition changes. The results demonstrated that 4-5 h was the best processing time, providing references for quality control and further study of Polygoni Multiflori Radix.

Simultaneous Determination and Comparative Pharmacokinetics of Fuzi Water-Soluble Alkaloids between Normal and Acute Heart Failure Rats by Ultra Performance Liquid Chromatography Method.

In order to compare the pharmacokinetics of Fuzi water-soluble alkaloids between normal and acute heart failure rats, an ultra performance liquid chromatography (UPLC) method for simultaneous determination of it in rat plasma was developed. Plasma samples were treated by protein precipitation method. Seven water-soluble alkaloids were separated on a CAPCELL column and detected under the optimized chromatography condition. The calibration curves of seven targeted components showed good linearity with r2 > 0.9990 with average recoveries from 82.72 to 103.33% and matrix effect ranged from 90.02 to 104.03%. The intra- and inter-batch relative standard deviations were <10.72%, and the relative error of accuracy was within the range of -12.79-4.44%, respectively. After oral administration, the pharmacokinetic characteristics of it were investigated. Compared with the normal group, the Cmax and AUClast (area under the plasma concentration-time curve from the time of dosing to the time of last quantifiable concentration) of seven components except 3-Methoxy-p-tyramine Hydrochloride were obviously increased with remarkable prolonged t1/2 in acute heart failure group. In conclusion, a rapid, simple and sensitive UPLC method for the simultaneous quantification of seven water-soluble alkaloids in rat plasma was developed and validated for the first time. And the study showed that the disease condition had an impact on pharmacokinetics of Fuzi water-soluble alkaloids in rats in vivo.

A New Strategy for Quality Evaluation and Identification of Representative Chemical Components in Polygonum multiflorum Thunb.

Polygonum multiflorum Thunb. (HSW) is widely used as herb medicine and health food additive. Recently, a series of HSW-induced hepatotoxicities have been reported and many studies have been carried out to investigate it. But contradictory conclusions were drawn that might be caused by the inconsistent quality of market decoction pieces. Therefore, the HSW decoction pieces quality was evaluated with a developed novel method in the paper. 25 batches of raw HSW (RHSW) and 21 batches of processed HSW (PHSW) samples were purchased from different provinces of China. HPLC determination was performed to identify and detect the contents of 16 chemical compounds in herbal material. Fingerprint similarity was analyzed using chromatography information and the results showed that most herbs were in good similarity. Then, a comprehensive evaluation strategy based on principal component analysis with representative quality control indicators was developed to evaluate the quality of HSW samples. And the rationality of the developed method was verified by HCA analysis. The results showed that the herb from Dabashan, Sichuan Province, no matter RHSW or PHSW had the best quality. Different representative components were selected for RHSW or PHSW decoction pieces which might be caused by the chemical reaction during processing. And most PHSW were unqualified according to the requirement of Chinese Pharmacopeia which might take the responsibility for the toxicity of HSW.

The Effects of Rheum palmatum L. on the Pharmacokinetic of Major Diterpene Alkaloids of Aconitum carmichaelii Debx. in Rats.

BACKGROUND AND OBJECTIVES: Aconitum carmichaelii Debx. (Fuzi) is usually compatible with Rheum palmatum L. (Dahuang) in clinic. The study is conducted to investigate the influence of Dahuang on the pharmacokinetics of Fuzi. METHODS: Twelve rats were randomly divided into two groups. Fuzi group was orally administered a single dose of 38.4 mg/kg total alkaloids from Fuzi, and Fuzi-Dahuang group was given 38.4 mg/kg total alkaloids from Fuzi and 76.8 mg/kg Dahuang anthraquinones at the same time. The plasma concentrations of aconitine (AC), mesaconitine (MC), and hypaconitine (HC), benzoylaconine (BAC), benzoylmesaconine (BMC), benzoylhypaconine (BHC), and aconine (ACN) were determined by ultra-performance liquid chromatography-quadrupole time of flight mass spectrometry method. The pharmacokinetic parameters were calculated including maximum plasma concentration (C max), area under the plasma concentration-time curve in all time-points (AUClast), apparent volume of distribution (V z/F), apparent plasma clearance (CL/F), elimination half-life (T 1/2), and time to achieve maximum concentration (T max). RESULTS: AUClast of diester diterpene alkaloids (DDAs) were 58.20, 169.78, 278.48 ng·h/mL for AC, MC, and HC in Fuzi-Dahuang group which were remarkably lower than that in Fuzi group (71.62, 183.13, 410.59 ng·h/mL for AC, MC, HC). CL/F was significantly increased from 173.88 to 218.85 mL/h for AC, 433.22 to 800.21 mL/h for MC, 1150.61 to 1307.30 mL/h for HC after combination. However, with the significantly increased C max, AUClast of monoester diterpene alkaloids (MDAs) and amine diterpenoid alkaloids (ADAs) were 152.42, 1238.95, 287.96, 123.33 ng·h/mL for BAC, BHC, BMC, ACN in Fuzi-Dahuang group which were remarkably higher than that in Fuzi group (54.47, 1105.48, 200.75, 86.48 ng·h/mL for BAC, BHC, BMC, ACN). At the same time, CL/F was significantly decreased from 1030.15 to 607.09, 3594.06 to 1437.54, 1441.23 to 1310.14, and 391.30 to 239.50 mL/h for each one after combination. CONCLUSIONS: Fuzi diterpene alkaloids pharmacokinetics was greatly influenced by Dahuang which may account for the compatibility mechanism of effect-enhancing and toxicity-reducing.

Development and Validation of an UPLC-Q-TOF-MS Method for Quantification of Fuziline in Beagle Dog After Intragastric and Intravenous Administration.

A specific and sensitive UPLC-Q-TOF-MS method operated in the positive ion mode was developed and validated for the quantification of Fuziline in Beagle dog plasma. Fuziline and Neoline internal standard were separated on an Acquity UPLC BEH C18 column with the total running time of 4 min using gradient elution at the flow rate of 0.25 mL/min. The calibration curves for Fuziline showed good linearity in the concentrations ranging from 2 to 400 ng/mL with correlation coefficients (r) greater than 0.9971. The lower limit of quantification was 0.8 ng/mL. Intra- and interbatch relative standard deviations ranged from 2.11 to 3.11% and 3.12 to 3.81%, respectively. Fuziline was stable under different sample storage and processing conditions. The developed method was successfully applied to the comparative pharmacokinetic study of Fuziline in Beagle dog after intravenous and oral administration. Low absolute bioavailability of Fuziline (1.45 ± 0.76%) suggested a significant metabolism transformation extent in Beagle dog.

The application of HPLC-MS/MS to studies of pharmacokinetics and interconversion of isoliquiritigenin and neoisoliquiritigenin in rats.

A specific and sensitive HPLC-MS/MS method was developed and validated for the simultaneously quantification of isoliquiritigenin (ISL) and neoisoliquiritin (NIS) in rat plasma by oral administration. Analytes were analyzed on an Agilent 6460 LC-MS/MS system (Agilent, USA) using an Agilent Zorbax SB-C18 column (4.6 × 150 mm, 5 µm). Gradient elution was applied for the analyte separation using a mobile phase composed of 0.1% formic acid aqueous solution and methanol at a flow rate of 1.0 mL/min with a total running time of 12 min. The calibration curves for ISL and NIS showed good linearity in the concentrations ranging from 0.001 to 4.000 µg/mL with correlation coefficients >0.998. The precision, accuracy, recovery and stability were deemed acceptable. The method was applied to the pharmacokinetics study of ISL and NIS in rats by single and combination administration. The result showed that Cmax and AUC0→t of ISL were markedly increased from 0.53 to 1.20 µg/mL, and from 69.63 to 200.74 min µg/mL by combination administration. The mean t1/2 value was also prolonged from 64.55 to 203.74 min in the combination group. These results indicated that NIS may have been metabolized to ISL which increased the absorption and extended the elimination of ISL. However, little difference was found for NIS pharmacokinetics parameters between single NIS and the combination group, which suggested that there was no significant biotransformation of ISL to NIS. Copyright © 2015 John Wiley & Sons, Ltd.

Pharmacokinetics and bioavailability study of neoline in Beagle dogs.

This paper is aim to investigate the pharmacokinetics and absolute bioavailability of neoline in Beagle dogs, and provide a theoretical basis for further study. Ethyl acetate was used for liquid-liquid extracting after 10% ammonia alkalizing. The method of UPLC-Q-TOF-MS was established for the determination of neoline plasma concentrations. Beagle dogs were orally or intravenously administered with neoline for pharmacokinetic and absolute bioavailability study. Good linear relationship of neoline was found over the range of 0.1-4 mg x L(-1) (R2 = 0.9982) and 2-100 microg x L(-1) (R2 = 0.9945). Intra-and inter-day precision, expressed as the relativestandard (RSD) were less than 5.0%. Accuracy, expressed as the relative error (RE) was within 90.0%-115%. The recovery of neoline in dog plasma was more than 80%. After 6 mg x kg(-1) for ig and 1 mg x kg(-1) for iv administration of neoline, the main pharmacokinetic parameters were analyzed with Winnonlin software. t(1/2) were (313.88 +/- 63.18), (236.33 +/- 229.84) min, and AUC(0-infinity) were (58,027.40 +/- 14,132.69), (473,578.02 +/- 82,333.08) min x microg x L(-1) for ig and iv administration respectively. The absolute bioavail ability was (73.15 +/- 10.29) %. The method of UPLC-Q-TOF-MS described in the report was sensitive, reliable and specific, and suitable for pharmacokinetic study of neoline in Beagle dog. The high absolute bioavailability of neoline in dog suggested good absorption of neline which was worth of further investigation.

The Influence of Aconitum carmichaelii Debx. on the Pharmacokinetic Characteristics of Main Components in Rheum palmatum L.

Rhei Radix et Rhizoma was one of the commonly used traditional Chinese medicines, and the compatibility of Rhei Radix et Rhizoma and Aconiti Lateralis Radix Praeparata was the basic herb pair applied in many Chinese traditional prescription. Rhubarb anthraquinones were the main bioactive materials of Rhei Radix et Rhizoma. To elucidate the compatibility of Rhei Radix et Rhizoma and Aconiti Lateralis Radix Praeparata, the pharmacokinetics of rhubarb anthraquinones as the main marker constituents were investigated. In the present study, pharmacokinetic differences of rhubarb anthraquinones were detected after oral administration of extract of Rheum palmatum L. and compatibility with Aconitum carmichaelii Debx. After oral administration, no difference of peak time can be found for anthraquinones between rhubarb group and compatibility group. But Cmax and area under the curve of aloe-emodin, emodin and chrysophanol in compatibility group were significantly higher than that in rhubarb group. Although the Cmax of rhein in compatibility group was much lower than that in rhubarb group, the area under the curve value was similar in two groups. The clearance and t1/2 of rhubarb anthraquinone were also changed after compatibility. The change of pharmacokinetics characteristics of rhubarb anthraquinone after compatibility may be caused by the drug-drug interaction medicated by chemical reaction and cytochromes P450.

The synergism mechanism of Rhubarb Anthraquinones on constipation elucidated by comparative pharmacokinetics of Rhubarb extract between normal and diseased rats.

In the study, it was hypothesized that Rhubarb Anthraquinones synergistically enhanced the purgative effect on constipation rat from the direct and indirect pathway at the same time. A validated HPLC method was successfully applied to elucidate the synergism mechanism from pharmacokinetics aspect after oral administration of Rhubarb extract with a dose of 0.25 g to normal and constipation rats. Comparison of the pharmacokinetic data of normal and constipation rats showed that there were significant differences (p < 0.05) in the main pharmacokinetic parameters. The C max and AUC of emodin in constipation rats were about ten times that of normal rats, while the t 1/2 was remarkably decreased (p < 0.05). However, a significant decrease (p < 0.05) in AUC value for aloe-emodin and rhein was observed in model group compared with normal group. The results may be attributed to the direct action of aloe-emodin and rhein on intestinal cell membranes and the indirect action of emodin on bowel movement through the adjustment by nervous system.

Development and validation of a UHPLC-qTOF-MS method for quantification of fuziline in rat plasma and its application in a pharmacokinetic study.

A specific and sensitive UHPLC-qTOF-MS method was developed and validated for quantification of fuziline in rat plasma after oral administration of three dosages. The analyte was separated on an Acquity UPLC BEH C18 column with a total running time of 3 min using a mobile phase of 0.1% formic acid aqueous solution and methanol (80:20, v/v) at a flow-rate of 0.25 mL/min. The calibration curves for fuziline showed good linearity in the concentrations ranging from 1 to 200 ng/mL with correlation coefficients >0.997. The precision, accuracy, recovery and stability were deemed acceptable. The method was applied to a pharmacokinetics study of fuziline in rats. The mean half-life was 5.93, 6.13 and 5.12 h for 1, 2 and 4 mg/kg oral administration of fuziline, respectively. The peak concentration and area under the concentration-time curve increased linearly with the doses. The sum of these results indicated that, in the range of the doses examined, the pharmacokinetics of fuziline in rat was based on first-order kinetics.

Genetic variants reducing MTR gene expression increase the risk of congenital heart disease in Han Chinese populations.

AIMS: Elevated homocysteine levels are known to be a risk factor for congenital heart disease (CHD), but the mechanism underlying this effect is unknown. During early embryonic development, homocysteine removal is dictated exclusively by the MTR activity. To examine the role of MTR in CHD risk, we identified genetic variants in MTR and investigated the mechanisms that affect its expression levels and that increase the risk of CHD in Chinese populations. METHODS AND RESULTS: The association between regulatory variants of the MTR gene and CHD was examined in three independent case-control studies in a total of 2340 patients with CHD and 2270 controls. The functional consequences of these variants were demonstrated using dual-luciferase assays, real-time polymerase chain reaction (PCR), electrophoretic mobility shift assays, surface plasma resonance, chromatin immunoprecipitation, and bisulfite sequencing, as well as by a group of predicted microRNAs using a gene reporter system. Two regulatory variants of MTR, -186T>G and +905G>A, were associated with an increased risk of CHD in both the separate and combined case-control studies (-186GG P = 1.32 × 10(-9); +905AA P = 6.35 × 10(-14)). Compared with the major allele, the -186G allele exhibited significantly lower promoter activity, decreased hnRNA and mRNA levels, reduced transcription factor binding affinity, and a more highly methylated promoter. The +905A allele exhibited a statistically stronger binding affinity to functional microRNAs that down regulate MTR expression at the translational level. Both of the minor alleles were correlated with elevated plasma homocysteine concentrations, indicating a genetic component for hyperhomocysteinaemia. CONCLUSIONS: Regulatory variants of the MTR gene increase CHD risk by reducing MTR expression and inducing the homocysteine accumulation and elevation.

[Study on the risk of HBV infection among spouses of HBsAg carriers].

OBJECTIVE: To investigate the risk of HBV infection among the spouses of hepatitis B virus surface antigen (HBsAg) carriers and to provide a reference for developing strategies on hepatitis B control and prevention. METHODS: A case-control study including HBsAg carriers aged 20 - 45 years-old from the nationwide sero-epidemiological survey for Hepatitis B in both Guangdong and Jiangxi provinces in 2006, together with their spouses were selected as case group, while. HBsAg negative persons and their spouses were among the control groups, under the same residential areas, gender, age and age of marriage to the HBsAg carriers. Questionnaire survey and hepatitis B serological markers detection were carried out, together with the HBV genotype detection among the HBsAg positive couples between husband and wife by PCR. RESULTS: Among the spouses of HBsAg carriers, the positive rate of HBsAg was 13.21%, while the rate was 6.29% for the spouse of HBsAg negative population, with difference statistically significant (χ² = 4.23, P < 0.05). HBsAg positive rate among spouses of the case group was higher than that in the control group. Among the spouses of HBsAg carriers, the HBsAg rate was positively correlated with the age of marriage, frequency of sexual intercourse and condom use. There were 21 pairs of HBsAg carriers between husband and wife, and HBV were isolated among 13 pairs, and there were 11 pairs carrying the same HBV genotype, accounting for 84.62%. HBV genotypes would include 8 pairs of type B and 3 pairs of type C. However, only 2 pairs were infected with different HBV genotype. CONCLUSION: High risks of HBV infection existed in the spouses of HBsAg carriers. It was important to ask the HBsAg carriers to take the initiative in informing their spouses, and carrying out the appropriate measures, such as safe sex or timely hepatitis B vaccination for the spouse of HBsAg carriers etc., so as to reduce the HBV transmission between husband and wife.

Determination of phillygenin in rat plasma by high-performance liquid chromatography and its application to pharmacokinetic studies.

The research group has been dedicated to the study of Fructus Forsythiae which was used widely in traditional Chinese medicines. And some research results have been accepted in Chinese Pharmacopeia. In a recent study, phillygenin was found to be a potential "metabolite" of phillyrin and the effective material of phillyrin may be changed according to the in vivo pharmacokinetic process. Therefore, a sensitive, specific, accurate, and reproducible reversed phase HPLC method for the determination of phillygenin in rat plasma was developed. Separation was achieved on a Hypersil ODS C18 column with UV detection at 277 nm. The good linear calibration curves ranged from 0.039 to 20 µg/mL with the limit of quantification estimated as 0.026 µg/mL. The intra- and inter-day precisions were in the range of 98-103 %. The average recoveries of phillygenin were 90.54, 92.47, and 92.15 % for phillygenin of 0.156, 1.25, and 10.0 µg/mL. And the Ruggedness of HPLC method was evaluated. The analytical method was also successfully applied to the pharmacokinetic study of phillygenin in rat for the first time. A rapid distribution was observed from the plasma concentration-time curves, and was followed by a quick elimination for phillygenin. The mean t 1/2z was 6.02, 5.62, and 5.79 min for 1.4, 2.8, and 5.6 mg/kg, respectively. The AUC (0-t) increased linearly from 166.29 to 332.48 mg/L min. All results indicated that, in the range of the doses examined, the pharmacokinetics of phillygenin in rat was based on first-order kinetics.

[Study on antibody response to revaccination of hepatitis B vaccine among firstly low-response adults].

OBJECTIVE: To evaluate and compare the antibody to hepatitis B virus (HBV) surface antigen (anti-HBs) response and the influent factors of revaccination of 4 kinds of hepatitis B vaccine (HepB) among firstly low-response adults. METHODS: A total of 11 590 adults who were 18 - 49 years old, never received HepB vaccination, without HBV infection history, HBs-Ag negative, and had been living at 3 towns of Zhangqiu county in Shandong province Ji'nan city for more than half a year, were selected in the study in July, 2009. Self-designed questionnaire was used to select the basic information of the subjects. The subjects were divided into 4 groups by cluster sampling, and were vaccinated according to the "0-1-6" immune procedure with 10 µg HepB made by recombinant deoxyribonucleic acid techniques in Saccharomyces Cerevisiae (HepB-SC), 10 µg HepB made by recombinant deoxyribonucleic acid techniques in Hansenula Polymorpha (HepB-HP), 20 µg HepB-SC and 20 µg HepB made by recombinant deoxyribonucleic acid techniques in Chinese hamster ovary cell (HepB-CHO), 3 doses respectively. The adults who were low-response to the primary hepatitis B vaccination (10 mU/ml ≤ anti-HBs < 100 mU/ml) were divided into four groups by cluster sampling. These groups were revaccinated with one-dose of above-mentioned four kinds of HepB respectively. Blood samples were drawn from each person one month after the revaccination. Anti-HBs was detected by chemiluminescence microparticle immunoassay and compared by the vaccine type. The influence factors about antibody response were also analyzed. RESULTS: Out of the 11 590 subjects, 8592 adults had accepted the primary vaccination of hepatitis B and been collected the blood samples; among whom, 1306 subjects showed low-response, at the rate of 15.20%. A total of 1034 low-response subjects accepted secondary strengthened vaccination and were collected blood samples; 55.13% of them showed anti-HBs seroconversion (anti-HBs ≥ 100 mU/ml); while the seroconversion rate in each group was 44.54% (106/238) in 10 µg HepB-SC group, 57.14% (156/273) in 10 µg HepB-HP group, 56.08% (143/255) in 20 µg HepB-SC group and 61.57% (165/268) in 20 µg HepB-CHO group, respectively. There was significant difference among the groups (χ² = 17.14, P < 0.01). The rates of anti-HBs seroconversion were significantly higher in 10 µg HepB-HP and 20 µg HepB-CHO groups than it in 10 µg HepB-SC group (χ² were 8.09 and 14.70 respectively, P < 0.01). The geometric mean concentration (GMC) of anti-HBs was 178.24 mU/ml among the low-responders after one dose of revaccination. The GMC was 109.77, 243.50, 144.98 and 242.83 mU/ml in 10 µg HepB-SC group, 10 µg HepB-HP group, 20 µg HepB-SC group and 20 µg HepB-CHO group, respectively. There was significant difference among groups (F = 9.52, P < 0.01). CONCLUSION: Anti-HBs response could be strengthened effectively after one-dose of HepB revaccination among the low-response adults. Many factors like the vaccine types could effect the immune effects to HepB. A better response could be achieved if the 20 µg HepB-CHO or 10 µg HepB-HP was used for revaccination.

[Comparison on the antibody response after primary immunization of 5 µg and 10 µg hepatitis B vaccine made by recombinant DNA techniques among newborns].

OBJECTIVE: To compare the antibody response induced by primary immunization with 5 µg and 10 µg hepatitis B vaccine made by recombinant DNA techniques among the newborns. METHODS: Healthy infants who had completed primary immunization with 5 µg hepatitis B vaccine made by recombinant dexyribonucleic acid techniques in Saccharomyces (Hep-SC) or 10 µg hepatitis B vaccine made by recombinant dexyribonucleic acid techniques in Hansenula polymorpha (HepB-HP) were included in the study. Kids under study were 7-12 months of age and had been on 0-1-6 schedule. Standardized questionnaire was used and blood samples were collected. The titer of antibody to hepatitis B surface antigen (anti-HBs) was detected by Chemiluminescence Microparticle Imunoassay (CMIA). If anti-HBs happened to be under 10 mIU/ml, HBV DNA was further detected by nested-PCR to distinguish occult hepatitis B virus infection. Sero-conversion rate and titer of anti-HBs were compared between the two kinds of hepatitis B vaccines. Multivariate analysis was used to find the relationship between the kind of hepatitis B vaccine as well as the antibody response after debugging the other influencing factors including month-age, gender, birth-weight, premature birth and mother's HBsAg status. RESULTS: 8947 infants vaccinated with 5 µg HepB-SC and 4576 infants vaccinated with 10 µg HepB-HP were investigated. In the 5 µg group, the rates of non-, low-, normal- and high-response were 1.88%, 15.18%, 61.42% and 21.52% respectively. In the 10 µg group, the corresponding rates were 0.15%, 2.16%, 29.42% and 68.26% respectively. The non-, low-, normal-response rates were all higher in 5 µg group than in 10 µg group (P<0.01), while the high-response rate was much higher in 10 µg group than in 5 µg group (P<0.01). The geometric mean concentration (GMC) of anti-HBs were 354.81 mIU/ml (95%CI: 338.84-363.08 mIU/ml) and 1778.28 mIU/ml (95%CI: 1698.24-1819.70 mIU/ml) in the 5 µg group and 10 µg group respectively. The GMC was statistically higher in the 10 µg group than in the 5 µg group (P<0.001). The sero-conversion rate and GMC were significantly different between the two groups even after debugging the other influencing factors. CONCLUSION: Better anti-HBs response could be achieved by primary immunization with 10 µg HepB-HP than with 5 µg HepB-SC among newborns.