AIM: Trigonelline (Tr) is the second most abundant alkaloid in coffee beans. This study developed an assay combining hydrophilic interaction chromatography with ultra performance liquid chromatography (HILIC-UPLC) for the quantification of Tr in rat plasma to determine its pharmacokinetic behavior. METHODS: After the administration of Tr by gavage as well as intravenous injection and that of methanol extract of coffee beans (MECB) orally, blood samples from the experimental rats were analyzed using the HILIC-UPLC assay. Pharmacokinetic parameters were determined using the standard non-compartmental method and calculated using Practical Pharmacokinetic Program Version 87/97. RESULTS: The HILIC-UPLC assay was validated with the linear range of 0.12-100 μg·mL(-1) and a lower limit of quantitation of 0.12 μg·mL(-1). Its accuracy, precision, recovery, and stability were within acceptable limits. The AUC(0-∞) (where AUC is the area under the plasma concentration-time curve) values were determined to be (4 066.83 ± 1 244.41) and (3 544.29 ± 908.80) min·μg·mL(-1) after Tr was orally and intravenously administered, respectively. It was (4 566.75 ± 1 435.64) min·μg·mL(-1) after MECB was orally administered. The absolute bioavailability of Tr alone reached 57.37%, whereas that of Tr in MECB was 64.42%. The relative bioavailability of the alkaloid was 112.29%. CONCLUSIONS The HILIC-UPLC assay for Tr determination is simple and accurate, and also exhibits good reproducibility. The bioavailability of stand-alone Tr and that of Tr in MECB were both good. Tr alone and that in MECB orally administered did not exhibit any significant difference.
Alkaloids ; blood ; pharmacokinetics ; Animals ; Chromatography, High Pressure Liquid ; methods ; Chromatography, Liquid ; methods ; Coffea ; chemistry ; Hydrophobic and Hydrophilic Interactions ; Male ; Plant Extracts ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo apply fluorescent real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in detecting influenza viruses.

METHODSA total of 207 oral swab samples were obtained in 16 collections from SARS patients and suspected influenza outbreak cases. They were subjected to influenza virus detection by fluorescent real-time RT-PCR, MDCK cell culture, and hemagglutinin inhibition assay.

RESULTSOut of 207 samples, 79 (38.16%) were positive for influenza viruses when tested by fluorescent real-time PCR, and 62 (29.95%) positive when tested by MDCK cell culture. There was a statistically significant difference between them (chi square=8.64, P less than 0.005). From 104 cases in 9 collections dual serum samples were obtainable. When tested with hemagglutinin inhibition assay, 64 cases (61.54%) showed a 4-fold increase against H3N2 antigen.

CONCLUSIONThis study showed that fluorescent real-time PCR is a reliable, sensitive, and fast method for detecting influenza viruses.

Cell Culture Techniques ; Humans ; Influenza A Virus, H3N2 Subtype ; isolation & purification ; Influenza A virus ; isolation & purification ; Influenza, Human ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; virology

BACKGROUNDSMAD proteins have recently been identified as the first family of putative transforming growth factor-beta1 (TGF-beta1) signal transducers. This study was to investigate the effects of TGF-beta1 and signal protein Smad3 on rat cardiac hypertrophy.

METHODSThe incorporation of [(3)H]-leucine was measured to determine the hypertrophy of cardiomyocyte incubated with different doses of TGF-beta1 in cultured neonatal cardiomyocytes. The model of rat cardiac hypertrophy was produced with constriction of the abdominal aorta. At different times after the operation, rats were killed, and their left ventricular mass index (LVMI) determined. The mRNA expression of TGF-beta1 and Smad3 of cultured cells and hypertrophic left ventricles were assessed by RT-PCR. The protein expression of Smad3 was assessed by Western blot.

RESULTSIn cultured neonatal cardiomyocytes, TGF-beta1 significantly promoted incorporation of [(3)H]-leucine. With the concentration of 3 pg/L, it increased the expression of Smad3 in mRNA and protein levels after 15 minutes, and continued for up to 8 hours of cultured cardiomyocytes. The LVMI and the expression of TGF-beta1 (mRNA) and Smad3 (mRNA and protein) of hypertrophic left ventricle were increased by day 3 after the operation and continued to the 4th week. The peak expression of these was in the second week after operation.

CONCLUSIONTGF-beta1 has positive effects on rat cardiomyocyte hypertrophy. Signal protein Smad3 could be related to the pathologic progression of rat cardiac hypertrophy.

Animals ; Aortic Coarctation ; metabolism ; Cardiomegaly ; etiology ; Cells, Cultured ; DNA-Binding Proteins ; physiology ; Leucine ; metabolism ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Smad3 Protein ; Trans-Activators ; physiology ; Transforming Growth Factor beta ; genetics ; physiology ; Transforming Growth Factor beta1

Objective::To investigate the processing purpose of Morindae Officinalis Radix (MO), Euodiae Fructus (EF) and Polygalae Radix (PR) processed by Glycyrrhizae Radix et Rhizoma (Gly). Method::The content of dapsone in rat plasma was determined by high performance liquid chromatography (HPLC), the mobile phase was acetonitrile (A)-water (B) for gradient elution (0-5 min, 10%-25%A; 5-20 min, 25%A) and detection wavelength was set at 292 nm. PK Solution 2.0 software was used to simulate pharmacokinetic parameters. Result::Within 300 min after dapsone was administrated, compared with the control (CTL) group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of MO (UMO) group. The elimination of dapsone was accelerated and its peak concentration (C_max) was decreased in the processed products of MO with Gly (GMO) groups, and they had positive correlation with proportion of Gly in GMO. Compared with the CTL group, the elimination of dapsone was slowed down, and its plasma concentration was increased and its peak time (T_max) was postponed in the unprocessed product of EF (UEF) group, while their C_max and T_max were changed in the processed products of EF with Gly (GEF) groups. Compared with the CTL group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of PR (UPR) group, while the elimination was accelerated and its plasma concentration was decreased in the processed products of PR with Gly (GPR) groups. Conclusion::The elimination of dapsone is slowed down in rats administered with UMO, UEF and UPR, while its elimination is accelerated in rats administered with the processed products of these three herbs with different proportions of Gly. Among the proportions, effect of processed products of these three herbs with 100∶6 (ratio of unprocessed product-Gly) on pharmacokinetics of dapsone is not significant.