The purpose of this study is to investigate the effects of multiple-trough sampling design and nonlinear mixed effect modeling (NONMEM) algorithm on the estimation of population and individual pharmacokinetic parameters. Oxcarbazepine and tacrolimus were used as one-compartment and two-compartment model drugs, respectively. Seven sampling designs were investigated using various number of trough concentrations per individual ranging from 1-4. Monte Carlo simulations were performed to produce state-steady trough concentrations. One-compartment model was used to fit simulated data from oxcarbazepine and tacrolimus. The accuracy and precision of the estimated parameters were evaluated using the median prediction error (PE), the median absolute PE and boxplot. The results indicated that trough concentrations could yield reliable estimates of apparent clearance (CL/F). For oxcarbazepine, as the number of trough concentrations per subject increased, the accuracy and precision of CL/F, between-subject variability (BSV) of CL/F and residual variability (RUV) tended to be improved. For tacrolimus, however, although no improvement were observed in the accuracy of CL/F and BSV of CL/F, the PE distribution ranges were significantly narrowed and the RUV estimates were less bias and imprecise. In terms of algorithm, Monte Carlo importance sampling (IMP) and IMP assisted by mode a posteriori estimation (IMPMAP) were consistently better than other methods. Additionally, the sampling design had no significant effects on the individual parameter estimates, which were only depended on the interaction between BSV and RUV in various algorithms. Decreased in BSV and RUV levels can improve the accuracy and precision of the estimation for both population and individual pharmacokinetic parameter estimates.
Algorithms ; Bayes Theorem ; Carbamazepine ; analogs & derivatives ; pharmacokinetics ; Humans ; Immunosuppressive Agents ; pharmacokinetics ; Models, Biological ; Monte Carlo Method ; Nonlinear Dynamics ; Regression Analysis ; Tacrolimus ; pharmacokinetics

Aged ; Castleman Disease ; complications ; Hepatitis B ; complications ; Humans ; Liver Neoplasms ; complications ; virology ; Lymphoma, Non-Hodgkin ; complications ; Male

Objective To investigate the effect and molecular mechanism of cyclooxygenase-2 (COX-2) on the benign prostatic hyperplasia (BPH) in rat model. Methods Thirty-six SD rats were randomly divided into 3 groups:normal control (group A,n=12),BPH model (group B,n=12) and BPH+selective COX-2 inhibitor celecoxib (group C,n=12). At the 5th week after treatment, the weight of the prostates was measured, and the morphological changes were examined under light microscope.Detection of ki-67 and TUNEL in prostatic epithelial and stromal cells was undertaken to assess the proliferation and apoptosis status.The protein and mRNA expression of COX-2,epidermal growth factor (EGF),basic fibroblast growth factor (bFGF) and transforming growth factor-?1 (TGF-?1) were analyzed by means of immunohistochemisty and RT-PCR. Results The prostate index [prostate wet weight (mg)/rat body weight (g)] of group B was significantly higher compared with those in groups A and C (1.88?0.17 vs 1.70?0.09 and 1.74?0.16,P0.05). Conclusions The increased expression of COX-2 may play an important role in the pathogenesis of BPH by modulating the expression of growth factors and affecting the proliferation and apoptosis of prostate cells.

0.05). Conclusions The expression of angiotensinⅡincreases in BPH tissues in rats. It is suggested that angiotensinⅡmay affect cell proliferation but not significantly affect cell apoptosis of the rat prostate.

objective To determine the existence of virulence-associated invA gene in different genospeeies of Leptospira interrogans reference strains in China.and to understand the alterations of invA gene transcription and expression of L.interrogans strain Lai before or after infecting cells.Methods PCR was applied to detect the invA gene of four L.interrogans strains belonging to four different genospecies and L.biflexa strain Patoc Ⅰ.The entire invA genes from the L.interrogans strains were cloned and then sequenced.The prokaryotic expression system of invA gene of L.interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai was constructed.Using Ni-NTA affinity chromatography,the target recombinant protein rInvA was extracted and purified.Rabbits were immunized with rInvA to obtain antiserum and the titer of antiserum was determined by immunodiffusion test.A model of L interrogans strain Lai infecting human embryo kidney cell line HEK293 was established to detect the alterations of invA gene transcription and expression of the leptospiral strain before or after infecting the host cells by real-time fluorescent quantitative RTPCR and western blot assay.Results All the four tested L.interrogans strains had invA gene whereas L.biflexa strain Patoc Ⅰ not.The similarity of nucleotide and putative amino acid sequences of invA genes from the four L.interrogans strains belonging four different genospecies were 99.33%-100%and 98.66%-100%,respectively.The constructed prokaryotic expression system could efficiently express rInvA and the immunodiffusion titer of rabbit anti-rInvA serum was 1:16.After L.interrogans strain Lai infecting HEK293 cells for 30 min or above,the microbe could adhere the surface of the cells.On the 30 min after the infection,the mRNA level of invA gene of L.interrogans strain Lai was remarkably upregulated,and on the 45 min after infection the mRNA level presented a peak value and then graduated decreased.On the 45 min or 60 min after L.interrogans strain Lai infecting HEK293 cells,InvA protein could be detectable but before infection or after infection for over 90 min the InvA protein expression was negative.Conclusion The invA gene is a unique gene for pathogenic L.interrogans.The invA gene of L interrogans has characteristics of cell-touched expression and transient expression,which may have a close correlation with adhering and invading host cells.

Objective To construct a plasmid DNA encoding G glycoprotein of respiratory syncytial virus(RSV) and investigate the protective immune response against RSV infection. Methods Recombinant plasmid DNA of pcDNA3.1~G was constructed by standard RT-PCR based cloning procedure. The immunogenicity of recombinant G protein transiently expressed in HEK293 cells was detected by Western blot. BABL/c mice were intramuscularly immunized with pcDNA3.1~G. Samples of lung, sera, bronchoalveolar lavage fluid(BALF) were collected before and after RSV challenge; virus titer in lung was detected by viral titration; sections of paraffin embedding lung tissues were stained by haematoxylin and eosin(HE) for histological analyses; sera anti-RSV IgG levels were examined by ELISA; Th1/Th2 cytokine were detected by ELISA kit, the T lymphocyte subsets of BALF was determined by immunefluorescence staining followed by flow cytometry. Results Plasmid DNA of pcDNA3.1~G was successfully constructed. The expressed target protein possesses immunogenicity. After challenge, pcDNA3.1~G immunized mice presented relieved pathological changes in lung as well as reduced lung viral titers. The RSV specific IgG was detected in sera of immunized mice. There was significantly increased number of CD25~+CD4~+ T cells in mice BALF. Conclusion We constructed a pcDNA3.1~G plasmid DNA vaccination which can induce evident protective cellular immunity against RSV infection in mice with the increased number of CD25~+CD4~+ T cell subpopulation.

To compare the difference of total phenol of magnolia solid dispersion prepared by different methods. Hot melt extrusion, solvent evaporation method, and fusion-cooling method were used to prepare total phenol of Magnolia accessory solid dispersion, Plastone S-630 and HPC. The drug dispersion state in the prepared solid dispersion was evaluated with DSC and X-ray diffraction; FT-IR method was used to analyze the possible connections between drug and accessories. Finally, accelerated stability-in vivo dissolution test was use to compare the stability differences between these three processes. The results of DSC and X-ray diffraction showed that all of the drug in solid dispersion processed by three processes can exist in amorphous form; FT-IR results also could not distinguish the difference between the three processes; accelerated stability-in vivo dissolution test showed the stability of solid dispersion prepared by HPC was better than Plastone S-630, and the same kinds of materials solid dispersion prepared by hot melt extrusion showed a better stability than the other two processes.
Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Magnolia ; chemistry ; Phenol ; chemistry ; Spectroscopy, Fourier Transform Infrared ; X-Ray Diffraction

HPLC analysis was performed to study the changes in chemical composition of ginseng extracts prepared from high quality ginseng with 0, 2, 4, 8 h of steamed times. An UFLC-MS/MS multiple-reaction monitoring (MRM) quantitative analysis was made to investigate the pharmacokinetic behavior differences of ginsenosides in mice ig administered of ginseng extracts with different steamed times in the negative ion mode, with Digoxin as the internal standard substance. The mice were injected with LPS to establish inflammation model after ig administration of ginseng for a week and the contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in mice plasma were detected by ELISA, in order to study on anti-inflammatory effects of ginseng with different steamed times. It was determined that levels of TNF-α and IL-1β were significantly decreased in inflammation model group ig administered of ginseng extracts with 8h of steamed time. The results showed that the chemical components in ginseng changed after steaming and the components into the blood changed, correspondingly. Ginseng with steamed 8 h contributes to anti-inflammatory effects. These results provided an experimental basis for revealing the active substance basis and dose-effect relationship of ginseng on anti-inflammatory effect.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacokinetics ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Ginsenosides ; chemistry ; pharmacokinetics ; Hot Temperature ; Humans ; Inflammation ; drug therapy ; Male ; Mice ; Mice, Inbred ICR ; Panax ; chemistry ; Time Factors

Objective To construct a recombinant lentivirus containing human beta defensins -3 ( hBD3 ) , connective tissue growth factor gene (CTGF) and enhanced green fluorescent protein (EGFP), and to detect its translation in rabbit bone marrow mesenchymal stem cells (BMSC).Methods The lentivirus containing hBD3, CTGF and EGFP genes was constructed in vitro.The titer of lentivirus was tested with end-paint dilution assay .Rabbit BMSCs were transfected with recombinant virus.The best value of multiplicity of infection (MOI) was tested.The expression condition, transfection efficacy and genetic stability of the target genes were evaluated by using fluorescence microscopy and flow cytometry . Western blotting was used to detect the expression of the target protein .Results Recombinant lentivirus vectors: Lenti-CTGF-hBD3-EGFP, Lenti-hBD3-EGFP, and Lenti-EGFP, were successfully obtained . The titer of the recombinant lentiviruses was 3.21 ×108, 5.80 ×108, and 1.16 ×109, respectively.The best MOI value to transfect BMSCs was 150. The transfection efficacy of these lentivirus vectors was high , reaching 79.72%as assessed by flow cytometry , and it could be stably inherited .Western blotting displayed that target protein expression was successful .Conclusion The construction of recombinant lentiviruses carrying hBD3 and CTGF genes is successful and can be effectively transfected into BMSCs .

Objective To determine the risk factors of ruptured hepatocellular carcinoma (HCC) and to study the prognostic factors of long-term survival.Methods Of the 4209 patients with HCC diagnosed and treated at the Eastern Hepatobiliary Surgery Hospital from Apr 2002 to Nov 2006,200 patients (4.8％) presented with ruptured HCC.These patients were studied retrospectively and the results of treatment were evaluated and compared with a randomly selected group of 202 patients who had no history of rupture and were treated during the study period.Results On multivariate logistic regression analysis,co-existing hypertension and cirrhosis,tumor size ＞5 cm,vascular thrombus and extrahepatic invasion were predictors of spontaneous rupture of HCC.For the 200 patients with spontaneous rupture of HCC,105 patients underwent elective one stage hepatic resection,33 received transcatheter arterial chemoembolization (TACE),and 62 were treated conservatively.The median survival time (MST) for patients with spontaneous rupture of HCC was 6 months (range,1-72 months),and the overall survival rates at 1-,3-and 5-year were 32.5％,10％ and 4％,respectively.The MST was 12 months (range,1-72 months) in the surgical group,4 months (range,1-30 months) in the TACE group and 1 month (range,0-19 months) in the conservative group.The 1-,3-and 5-year overall survival rates in patients with ruptured HCC who received partial hepatectomy were 57.1 ％,19.0％ and 7.6％,respectively,compared with 77.1％,59.8％ and 41.2％ in 98 patients who underwent partial hepatectomy for HCC without rupture (P＜0.001).Conclusions For patients with HCC who had underlying of hypertension and cirrhosis,extrahepatic invasion and tumor size ＞5 cm,there was a high propensity to rupture.Prolonged survival could be achieved in selected patients who received one-stage partial hepatectomy,although the survival results were inferior to the patients had no rupture.