Objective To evaluate the pharmacokinetics(PK)and pharmacodynamics(PD)of propofol injectable emulsion,and to assess the bioequivalence of test and reference formulations in healthy Chinese adult volunteers.Methods Thirty-two healthy Chinese adult volunteers were recruited and randomly assigned to a fasting.single-dose,two-period and double-crossover study.Propofol was given to eligible subjects at a speed of 30 μg·kg-1·min-1 for 30 min.The concentration of propofol in plasma was determined by avalidated high performance liquid chromatography-tandem mass spectrometery(HPLC-MS/MS)method.The PK parameters of the two preparations were calculated.Bispectral index(BIS)was measured to calculate the PD parameters of two formulations.Adverse events during the trial were recorded.Results Thirty-one volunteers were included in the pharmacokinetic parameter set.The mean values of PK parameters of test andreference formulations were as follows:Cmax were(660.87±110.25)and(683.13±125.75)ng·mL-1;AUC0-t were(473.50±86.03)and(478.40±80.25)h·ng·mL-1;AUC0-∞ were(500.45±96.49)and(507.84±88.00)h·ng·mL-1;tmax were 0.47(0.25,0.53)and 0.50(0.40,0.54)h;t1/2 were(2.97±1.74)and(3.08±1.82)h.Thirty-one volunteers were included in the bioequivalence set.The 90％confidence intervals(CI)for the geometric mean ratios of Cmax,AUC0-t,AUC0-∞ were 92.64％-101.39％,96.43％-101.00％,95.67％-100.70％,respectively.The mean values of PD parameters of test and reference formulations were as follows:BISmin were(75.94±13.66)and(74.39±12.32);BISAUC0-60minwere 5 569.85±182.78 and 5 575.68±166.19;T-BISmin were 23.00 and 29.00 min,respectively.There were no serious adverse events.Conclusion Two formulations of propofol injectable emulsion were bioequivalent and both of them exhibited good safety.

Objective To evaluate the pharmacokinetics(PK)and pharmacodynamics(PD)of propofol injectable emulsion,and to assess the bioequivalence of test and reference formulations in healthy Chinese adult volunteers.Methods Thirty-two healthy Chinese adult volunteers were recruited and randomly assigned to a fasting.single-dose,two-period and double-crossover study.Propofol was given to eligible subjects at a speed of 30 μg·kg-1·min-1 for 30 min.The concentration of propofol in plasma was determined by avalidated high performance liquid chromatography-tandem mass spectrometery(HPLC-MS/MS)method.The PK parameters of the two preparations were calculated.Bispectral index(BIS)was measured to calculate the PD parameters of two formulations.Adverse events during the trial were recorded.Results Thirty-one volunteers were included in the pharmacokinetic parameter set.The mean values of PK parameters of test andreference formulations were as follows:Cmax were(660.87±110.25)and(683.13±125.75)ng·mL-1;AUC0-t were(473.50±86.03)and(478.40±80.25)h·ng·mL-1;AUC0-∞ were(500.45±96.49)and(507.84±88.00)h·ng·mL-1;tmax were 0.47(0.25,0.53)and 0.50(0.40,0.54)h;t1/2 were(2.97±1.74)and(3.08±1.82)h.Thirty-one volunteers were included in the bioequivalence set.The 90％confidence intervals(CI)for the geometric mean ratios of Cmax,AUC0-t,AUC0-∞ were 92.64％-101.39％,96.43％-101.00％,95.67％-100.70％,respectively.The mean values of PD parameters of test and reference formulations were as follows:BISmin were(75.94±13.66)and(74.39±12.32);BISAUC0-60minwere 5 569.85±182.78 and 5 575.68±166.19;T-BISmin were 23.00 and 29.00 min,respectively.There were no serious adverse events.Conclusion Two formulations of propofol injectable emulsion were bioequivalent and both of them exhibited good safety.

Objective To evaluate the bioequivalence and safety of ezetimibe tablets in healthy Chinese subjects.Methods The study was designed as a single-center,randomized,open-label,two-period,two-way crossover,single-dose trail.Subjects who met the enrollment criteria were randomized into fasting administration group and postprandial administration group and received a single oral dose of 10 mg of the subject presparation of ezetimibe tablets or the reference presparation per cycle.The blood concentrations of ezetimibe and ezetimibe-glucuronide conjugate were measured by high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS),and the bioequivalence of the 2 preparations was evaluated using the WinNonlin 7.0 software.Pharmacokinetic parameters were calculated to evaluate the bioequivalence of the 2 preparations.The occurrence of all adverse events was also recorded to evaluate the safety.Results The main pharmacokinetic parameters of total ezetimibe in the plasma of the test and the reference after a single fasted administration:Cmax were(118.79±35.30)and(180.79±51.78)nmol·mL-1;tmax were 1.40 and 1.04 h;t1/2 were(15.33±5.57)and(17.38±7.24)h;AUC0-t were(1 523.90±371.21)and(1 690.99±553.40)nmol·mL-1·h;AUC0-∞ were(1 608.70±441.28),(1 807.15±630.00)nmol·mL-1·h.The main pharmacokinetic parameters of total ezetimibe in plasma of test and reference after a single meal:Cmax were(269.18±82.94)and(273.93±87.78)nmol·mL-1;Tmax were 1.15 and 1.08 h;t1/2 were(22.53±16.33)and(16.02±5.84)h;AUC0_twere(1 463.37±366.03),(1 263.96±271.01)nmol·mL-1·h;AUC0-∞ were(1 639.01±466.53),(1 349.97±281.39)nmol·mL-1·h.The main pharmacokinetic parameters Cmax,AUC0-tand AUC0-∞ of the two preparations were analyzed by variance analysis after logarithmic transformation.In the fasting administration group,the 90％CI of the log-transformed geometric mean ratios were within the bioequivalent range for the remaining parameters in the fasting dosing group,except for the Cmax of ezetimibe and total ezetimibe,which were below the lower bioequivalent range.The Cmax of ezetimibe,ezetimibe-glucuronide,and total ezetimibe in the postprandial dosing group was within the equivalence range,and the 90％CI of the remaining parameters were not within the equivalence range for bioequivalence.Conclusion This test can not determine whether the test preparation and the reference preparation of ezetimibe tablets have bioequivalence,and further clinical trials are needed to verify it.

This study investigated the effect of puerarin on human umbilical vein endothelial cells (HUVEC) injured with hydrogen peroxide (H₂O₂). HUVEC were divided into three groups: a control group, a model group (H₂O₂ 400 μmol·L^-1) and a puerarin-treated group (3, 10, 30 and 100 μmol·L^-1). HUVEC were cultured with varied concentration of puerarin for 2 h and treated with H₂O₂ for another 24 h. Cell proliferation was detected by a CCK-8 assay. The mitochondrial membrane potential was measured by a JC-1 fluorescent probe. A transwell chamber assay was adopted to observe cell migration ability. Mitochondrial respiratory function was measured in a two-chamber titration injection respirometer (Oxygraph-2k). The expression of interleukin-1β (IL-1β), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) was detected by quantitative real-time PCR. The expression of pyroptosis-mediated proteins, including cleaved-cysteinyl aspartate-specific proteinase-1 (caspase-1), N-gasdermin D (N-GSDMD), NOD-like receptor protein 3 (NLRP3) and purinergic ligand-gated ion channel 7 receptor (P2X7R) was detected by Western blot. The results show that 400 μmol·L^-1 H₂O₂ treatment for 24 h causes obvious damage to HUVEC. Compared with the model group, puerarin protected against cellular injury in a dose-dependent manner, with the greatest effect at a dose of 30 and 100 μmol·L^-1. Puerarin significantly decreased the mitochondrial membrane potential and improved mitochondrial function. Puerarin inhibited cell migration induced by H₂O₂, suppressed the expression of IL-1β, IL-18 and TNF-α, and down-regulated the pyroptosis-mediated protein. These changes are statistically significant (P < 0.05). These findings demonstrate that puerarin has a protective effect against H₂O₂-induced oxidative damage of HUVEC by inhibiting the migration of HUVEC cells. The mechanism may be related to improved mitochondrial respiratory function and inhibition of pyroptosis.

OBJECTIVETo investigate the distribution and change of the causes of fever of unknown origin(FUO).

METHODSThe clinical data of 500 inpatients with FUO in our center between December 2003 and June 2014 were retrospectively analyzed. The diagnostic methods,etiologies,and their possible relationship with age,sex,fever duration,and period.

RESULTSOf these 500 FUO patients,452(90.4%)were confirmed to be with fever caused by conditions including infectious diseases [(n=231,46.2%;e.g.tuberculosis(32.9%,76/231)],connective tissue diseases(CTD)(n=99,19.8%),neoplasms(n=58,11.6%),miscellaneous causes(n=64,12.8%). The causes were not identified in 48 cases(9.6%).The proportion of CTD in female patients was significantly higher than that in male patients(26.3% vs. 14.5%,P=0.025),whereas the proportion of neoplasms in male patients was significantly higher than that in female patients(14.5% vs. 8.0%,P=0.001). Infectious diseases was the most common cause in all age groups,CTD ranked the second in the 21-39-year group and 40-59-year group,and neoplasm was the second most coomon cause in the over 60 year group. Thus,the distribution of FUO etiologies significantly differed in different age groups(χ(2)=43.10,P=0.000). The duration of fever in patients with neoplasms [60(28,90)d] was longer than that in patients with infectious diseases [28(21,42)d,Z=-4.168,P=0.000] or CTD [30(21,60)d,Z=-2.406,P=0.016)]. Compared with the level in 2003-2008,the proportion of CTD significantly increased in 2009-2014(13.7% vs. 23.8%,χ(2)=8.598,P=0.003),along with the dicrease of the proportions of infectious diseases,neoplasms and miscellaneous diseases were decreased(all P>0.05).

CONCLUSIONSInfectious diseases(in particular,tuberculosis)remains the major cause of FUO. CTD and neoplasms also play important roles in the development of FUO. The distributions of the FUO etiologies have certain differences in terms of age,sex,duration of fever,and period.

Connective Tissue Diseases ; Female ; Fever of Unknown Origin ; Humans ; Male ; Neoplasms ; Retrospective Studies ; Tuberculosis

AIMS: To develop an HPLC-MS/MS method for the quantification of platycodin D (PD) in rat plasma, and to acquire the main pharmacokinetic parameters of PD after oral administration of pure PD or of Platycodi Radix extract (PRE) containing PD. METHOD: Plasma samples were pretreated with solid-phase extraction using an Oasis® HLB SPE cartridge. Madecassoside was used as the internal standard (IS). Chromatographic separation was achieved on an ODS column (100 mm × 2.1 mm i.d., 3.5 μm) with a mobile phase consisting of acetonitrile/water (30 : 70, V/V) containing 0.1 mmol·L(-1) ammonium acetate at a flow rate of 0.25 mL·min(-1). The detection was performed on a triple quadruple tandem mass spectrometer using an electrospray ionization (ESI) source with a chromatographic run time of 3.0 min. The detection was operated by multiple reaction monitoring (MRM) of the transitions of m/z 1 223.6→469.2 for PD and of m/z 973.6→469.2 for madecassoside (IS), respectively. RESULTS: The calibration curve was linear from 5 to 2 000 ng·mL(-1) (r(2) >0.99) with a lower limit of quantification (LLOQ) of 5 ng·mL(-1). The intra- and inter-day precision (relative standard deviation, RSD) values were below 15% and the accuracy (relative error, RE) was from -15% to +15% at three quality control (QC) levels. Plasma concentrations of PD were determined for 24 h after i.v. administration of PD, and oral administration of PD and PRE, respectively. The absolute oral bioavailability of PD in rats was found to be (0.48 ± 0.19)% when administered PD, and to be (1.81 ± 0.89)% when administered PRE. CONCLUSION The developed HPLC-MS/MS method was successfully applied to assess the pharmacokinetic parameters and oral bioavailability of PD in rats after administration of PD and Platycodi Radix extract.
Administration, Oral ; Animals ; Biological Availability ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Male ; Plant Roots ; chemistry ; Platycodon ; chemistry ; Rats ; Rats, Sprague-Dawley ; Saponins ; blood ; pharmacokinetics ; Tandem Mass Spectrometry ; methods ; Triterpenes ; blood ; pharmacokinetics

Objective To explore the effects and related mechanisms of cilostazol on rat vascular smooth muscle cells (VSMCs)proliferation.Methods VSMCs were treated with DMEM (control) and various doses of cilostazol (1.0 × 10-7,2.5 × 10-7,5.0 × 10-7,7.5 × 10-7 and 1.0 × 10-6 mol/L) for 13 d (cell counting) or 72 h.Proliferation of VSMCs was investigated by cell-counting,MTT and flow cytometry analysis.Cell apoptosis was determined by TUNEL staining.mRNA and protein expressions of cell cycle regulatory proteins,such as Rb,p53 and p21 were detected by RT-PCR and Western blot,respectively.Results Cilostazol inhibited VSMCs proliferation and induced VSMCs arrest at G1 phase in a dose-dependent manner.High dose of cilostazol (7.5 × 10-7 and 1.0 × 10-6 mol/L) induced VSMCs apoptosis.p53 mRNA expression in 2.5 × 10-7 mol/L to 7.5 × 10-7 mol/L groups as well as 1.0 × 10-6 mol/L group (3.22 ±0.45 vs.1.75 ±0.32) and p53 protein expression in 7.5 × 10-7 mol/L group and 1.0 × 10-6 mol/L group (0.53 ± 0.11 vs.0.18 ± 0.06) were significantly upregulated after 72 h culture (all P＜0.05 vs.control).Low dose of cilostazol (1.0 × 10-7,2.5 × 10-7 and 5.0 × 10-7 mol/L)significantly upregulated p21 mRNA expression compared to control group (1.86 ± 0.19,2.20 ± 0.24 and 2.10 ± 0.18 vs.1.210 ± 0.18,all P ＜ 0.05).Similarly,Rb mRNA expression was significantly upregulated in 1.0 × 10-7,2.5 × 10-7 and 5.0 × 10 7 mol/L groups (0.89 ±0.07 vs.0.38 ±0.04)compared with control group (all P ＜ 0.05).However,high dose cilostazol (7.5 × 10-7 and 1.0 × 10-6mol/L) significantly downregulated p21 mRNA expression (0.81 ±0.09 vs.1.21 ±0.18,0.36 ±0.10 vs.1.2t ±0.18,all P ＜0.05 vs.control) and Rb mRNA expression (0.12 ±0.02 and 0.11 ±0.02 vs.0.38± ± 0.04,all P ＜ 0.05 vs.control).p21 and Rb protein expressions also upregulated at low concentrations of cilostazol and downregulated at high concentrations of cilostazol.Conclusion Cilostazol could inhibit the proliferation of rat VSMCs through modulating Rb-p53-p21 pathway and induce VSMCs apoptosis through upregulating p53.

To construct a recombinant baculovirus expressing glycoprotein (GP) of RV SRV9 strain and test the immunological efficacy in mice, open reading frame of rabies virus GP gene of SRV9 strain was cloned into the shuttle vector Bacmid to construct the recombinant shuttle plasmid Bacmid-G and transfection was performed into S f9 cells with the recombinant shuttle plasmid. CPE appeared in cell cultures was identified by electronmicroscopy. Western-blot, IFA and immunity tests in mice were performed to identify the immunoreactivity and immunogenicity of the expression products. Our results showed a recombinant baculovirus expressing GP protein of rabies virus SRV9 was obtained. The expression products possessed a favorable immunogenicity and fall immunized mice could develop 100% protective level of anti-rabies neutralizing antibody. In conclusion, The SRV9 glycoprotein expressed by the recombinant baculovirus in this study had good immunogenicity and could induce anti-rabies neutralizing antibody, which laid the foundation of further development of rabies subunit vaccine.
Animals ; Antibodies, Viral ; immunology ; Baculoviridae ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Glycoproteins ; administration & dosage ; genetics ; immunology ; Humans ; Mice ; Rabies ; immunology ; prevention & control ; virology ; Rabies Vaccines ; administration & dosage ; genetics ; immunology ; Rabies virus ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology

OBJECTIVETo observe the long-term effect of plasma-derived HBV vaccine.

METHODSThe effect of a plasma-derived HBV vaccine which was given to children born in 1986 in Huangpu district in Shanghai were followed up once every two years and testing for HBsAg, anti-HBs and anti-HBc was carried out. Compared to background results from cross-sectional survey of hepatitis B virus in 1984 and 1985 (as internal control) as well as finding of survey targeted in non-plasma-derived HBV vaccine of children born in the same time in the nearby area from results investigated in 1991 (as external control), positive rate was calculated to assess the effect of protection.

RESULTSThe population immunized was followed up for 23 years and 5993 blood samples were collected. During the period of follow-up, the positive rate of anti-HBs decreased from 89.01% to 18.77% and the average level was 40.39%. The average positive rate of anti-HBc was 1.87%. The annual positive rate fluctuated around the average level. HBsAg positive rate remained less than 1.00% (0.46% - 0.98%), with an average of 0.62% (37/5993). Ranges of positive efficacy were from 81.37% to 95.78% against background control and 72.76% against external control.

CONCLUSIONThe plasma-derived HBV vaccine showed a good long-term protective effect and there was no need for boosting the immunization 23 years later.

China ; epidemiology ; Female ; Follow-Up Studies ; Hepatitis B ; epidemiology ; prevention & control ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; immunology ; therapeutic use ; Humans ; Immunization Programs ; Infant, Newborn ; Male ; Vaccination

Objective To observe the long-term effect of plasma-derived HBV vaccine.Methods The effect of a plasma-derived HBV vaccine which was given to children born in 1986 in Huangpu district in Shanghai were followed up once every two years and testing for HBsAg,anti-HBs and anti-HBc was carried out.Compared to background results from cross-sectional survey of hepatitis B virus in 1984 and 1985 (as internal control) as well as finding of survey targeted in non-plasma-derived HBV vaccine of children born in the same time in the nearby area from results investigated in 1991 (as external control),positive rate was calculated to assess the effect of protection.Results The population immunized was followed up for 23 years and 5993 blood samples were collected.During the period of follow-up,the positive rate of anti-HBs decreased from 89.01％ to 18.77％ and the average level was 40.39％.The average positive rate of anti-HBc was 1.87％.The annual positive rate fluctuated around the average level.HBsAg positive rate remained less than 1.00％ (0.46％-0.98％),with an average of 0.62％ (37/5993).Ranges of positive efficacy were from 81.37％ to 95.78％ against background control and 72.76％ against external control.Conclusion The plasma-derived HBV vaccine showed a good long-term protective effect and there was no need for boosting the immunization 23 years later.