BACKGROUND/AIMS: Anti-viral therapy using hepatitis B immune globulin and lamivudine could not prevent HBV recurrence after liver transplantation (LT) completely. Adefovir dipivoxil is a acyclic nucleotide phosphate analogue and known to have potent anti-HBV effect. In this study, we analyzed the therapeutic effect of adefovir for recurrent or de novo HBV infection after LT. METHODS: From December 2002 to October 2004, adefovir was administered in 12 post-LT patients of HBV infection (11 recurrent and 1 de novo infection). In these patients, lamivudine and other combined therapies were used before the introduction of adefovir. Thereafter, adefovir combined with lamivudine was administered to all patients. RESULTS: The duration of adefovir administration was 5.5-18 (median, 15.5) months. The median values of serum AST and ALT levels were significantly reduced from 86+/-80 IU/L and 140+/-103 IU/L, respectively before the adefovir administration to 42+/-19 IU/L and 38+/-33 IU/L after 2 months of administration. This trend of improved liver function persisted throughout the follow-up period. HBeAg seroconversion was achieved in 4 of 10 patients (40%) and HBsAg seroconversion was observed in 1 of 10 patients (10%). HBV DNA levels have decreased to undetectable levels by hybridization assay in 6 of 7 patients within the first 2 months of therapy. Nephrotoxicity and hypophosphatemia were not found in all of these patients. CONCLUSIONS: Based on this preliminary result, adefovir dipivoxil seems to be an effective and safe antiviral agent leading to viral inhibition and clinical improvement in post-LT patients with recurrent or de novo HBV infection.
Adenine/administration & dosage/*analogs & derivatives ; Adult ; Antiviral Agents/*administration & dosage ; Drug Therapy, Combination ; English Abstract ; Female ; Hepatitis B, Chronic/*drug therapy ; Humans ; Lamivudine/administration & dosage ; *Liver Transplantation ; Male ; Middle Aged ; Phosphonic Acids/*administration & dosage ; Recurrence ; Reverse Transcriptase Inhibitors/administration & dosage

A simple, rapid and sensitive method was developed for the quantification of tenofovir in plasma of Beagle dogs using HPLC-MS/MS analysis. The analytes tenofovir and internal standard (IS) adefovir were separated on a Zorbax SB-C18 column (3.5 microm, 100 mm x 2.1 mm, Agilent, USA) with mobile phase of methanol/water containing 0.3% formic acid using a gradient elution mode at a flow rate of 0.2 mL x min(-1). The plasma sample preparation was a simple deproteinization by the addition of 20% trichloroacetic acid followed by centrifugation. The detection was performed in positive selected reaction monitoring (SRM) mode with an electrospray ionization (ESI) source. The reactions monitored were m/z 288.1-176.2 for tenofovir and m/z 274.1-162.2 for adefovir (IS). Linear detection responses were obtained for tenofovir ranging from 10 to 5 000 ng x mL(-1). The intra- and inter-day precisions (RSD%) was no more than 6.3% with high recovery and good stability for the quantification, indicating the present method was specific, fast, accurate and reliable. The method was successfully applied to the pharmacokinetic study of two tenofovir agents. Tenofovir dipivoxil fumarate (BP0018, test agent) and tenofovir disoproxil fumarate (reference agent) were orally administrated to 8 Beagle dogs according to the 2 x 2 crossover design. Comparing with the reference agent, the longer MRT and t1/2 were obtained in the group of BP0018, while no significant difference was observed in AUC(0-t), AUC(0-infinity), C(max) and t(max) between them, suggesting that tenofovir dipivoxil fumarate was bioequivalent to the tenofovir disoproxil fumarate in Beagle dogs.
Adenine ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics ; Administration, Oral ; Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; Dogs ; Female ; Male ; Organophosphonates ; administration & dosage ; blood ; pharmacokinetics ; Phosphorous Acids ; administration & dosage ; pharmacokinetics ; Reverse Transcriptase Inhibitors ; blood ; Tandem Mass Spectrometry ; Tenofovir

BACKGROUNDCapsular contracture has become the most common complication associated with breast implant. Transforming growth factor-beta (TGF-β) is well known for a prominent role in fibrotic diseases. Due to the critical role of TGF-β in pathogenesis of capsular formation, we utilized thermosensitive C/GP hydrogel to controlled release of TGF-β receptor kinase inhibitor (SD208) and investigated their effects on capsular contracture.

METHODSIn vitro degradation and drug release of C/GP hydrogel were performed. Twenty-four rabbits underwent subpanniculus implantation with 30 ml smooth silicone implants and were randomly divided into four groups as follows: Group 1 received saline solution; Group 2 received SD208; Group 3 received SD208-C/GP; Group 4 received C/GP. At 8 weeks, the samples of capsular tissues were analyzed by hematoxylin and eosin and immunohistological staining. The mRNA expression of collagen III and TGF-β1 was detected by RT-PCR assay.

RESULTSC/GP hydrogel could be applied as an ideal drug delivery vehicle which supported the controlled release of SD208. SD208-C/GP treatment showed a significant reduction in capsule thickness with fewer vessels. The histological findings confirmed that the lower amounts of inflammatory cells and fibroblasts infiltrate in SD208-C/GP group. In contrast, typical capsules with more vessel predominance were developed in control group. We did not observe the same inhibitory effect of SD208 or C/GP treatment on capsular contracture. Moreover, SD208-C/GP therapy yielded an evident down-regulation of collagen III and TGF-β1 mRNA expression.

CONCLUSIONSThis study demonstrated that controlled release of TGF-β receptor kinase inhibitor from thermosensitive C/GP hydrogel could significantly prevent capsule formation after mammary implants.

Animals ; Breast Implantation ; adverse effects ; Chitosan ; chemistry ; Glycerophosphates ; chemistry ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Immunohistochemistry ; Protein Kinase Inhibitors ; administration & dosage ; therapeutic use ; Rabbits ; Receptors, Transforming Growth Factor beta ; antagonists & inhibitors ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVESTo investigate the pharmacological effects of azidothymidine (AZT) on p33ING1b expression, senescence and apoptosis of TJ905 glioblastoma cells.

METHODSTJ905 cells were treated with AZT at a serial concentrations of 50, 100 and 200 µmol/L. Semi-quantitative RT-PCR and cytochemical staining of senescence related-galactosidase (sβ-Gal) were used to evaluate the expression of p33ING1b mRNA and to label the senescent cells at the 1st, 3rd and 6th generations, respectively. In situ cell death detection and single cell gel electrophoresis were used to detect the apoptosis at the 3rd and 6th generations.

RESULTSAZT induced the expression of p33ING1b mRNA and senescence of the tumor cells of the 1st generation in a dosage and time dependent manner. At the 6th generation, the relative amount of p33ING1b RT-PCR product (1.44±0.23) and sβ-Gal labeling index of 200 µmol/L group (45.62±6.74) were significantly higher than those of the 1st (0.95±0.13 and 7.82±2.40) and the 3rd generation cells (1.35±0.23, 26.27±7.17) of the same group, and cells of the same generation in the 50 µmol/L (0.85±0.24, 27.37±6.41) and 100 µmol/L groups (1.23±0.34, 35.49±5.12, P<0.01). There was a significant positive correlation between the p33ING1b mRNA expression and the labeling index of sβ-Gal. Pro-apoptotic effects of AZT became obvious at the 6th generation.

CONCLUSIONAZT upregulates the expression of p33ING1b, a possible mechanism in regulating senescence and apoptosis of the TJ905 cells.

Apoptosis ; drug effects ; Brain Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cellular Senescence ; drug effects ; Dose-Response Relationship, Drug ; Glioblastoma ; metabolism ; pathology ; Humans ; Inhibitor of Growth Protein 1 ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Reverse Transcriptase Inhibitors ; administration & dosage ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Proteins ; genetics ; metabolism ; Zidovudine ; administration & dosage ; pharmacology

BACKGROUND/AIMS: The therapeutic strategies of applying adefovir for treating lamivudine resistant HBV mutants are controversial. Thus, we observed the clinical outcomes after discontinuation of lamivudine to establish the timing to initiate adefovir therapy. METHODS: Fifty chronic hepatitis B (CHB) patients with lamivudine resistant HBV mutants who had received lamivudine for more than 12 months were included in the study. We investigated the clinical outcomes at 6 months after the end of treatment (EOT). We compared the serial clinical outcomes among respective groups based on serum ALT at the EOT and the clinical characteristics of patients with or without acute exacerbation (AE) and the HBeAg loss. We also investigated the predictive parameters of AE and HBeAg loss. RESULTS: Fifteen patients (30%) had experienced AE at 6 months after the EOT. Four patients received antiviral agents because of their hepatic decompensation. Patients with AE had higher serum ALT values and lower HBV DNA titers at EOT compared with those patients without AE. Serum ALT at the EOT was the predictive parameter of AE. Eight patients (21.6%) had newly developed HBeAg loss at 6 months after EOT. The total bilirubin at EOT was the predictive parameter of HBeAg loss. CONCLUSIONS: CHB patients with lamivudine resistant HBV mutants had favorable clinical outcomes at 6 months after EOT. Therefore, we can consider observing the clinical courses after discontinuation of lamivudine and it is not always required to overlap the adefovir for treating lamivudine resistant HBV mutants except for the treatment of patients with a high risk of developing decompensation.
Adenine/administration & dosage/analogs & derivatives ; Adult ; Antiviral Agents/*administration & dosage ; *Drug Resistance, Viral ; English Abstract ; Female ; Hepatitis B e Antigens/blood ; Hepatitis B virus/drug effects ; Hepatitis B, Chronic/*drug therapy/virology ; Humans ; Lamivudine/*administration & dosage ; Male ; Middle Aged ; Phosphonic Acids/administration & dosage ; Reverse Transcriptase Inhibitors/*therapeutic use ; Treatment Outcome

OBJECTIVETo explore the strategy for the treatment of chronic hepatitis B with YMDD mutation.

METHODSA total of 120 chronic hepatitis B patients with YMDD mutation were randomly assigned into four groups. In group A, patients received adefovir dipivoxil for 48 weeks. In group B, patients received adefovir dipivoxil in combination with lamivudine during the first 12 weeks and adefovir dipivoxil only for the following 36 weeks. In group C, patients received adefovir dipivoxil in combination with lamivudine for 48 weeks. In group D, patients received entecavir for 48 weeks.

RESULTSThe rate of rebound of alanine aminotransferase (ALT) was 30.0% (9/30), 10.0% (3/30), 6.7% (2/30), 10.0% (3/30) (P < 0.05) during the first 12 weeks, and one patient with severe hepatitis was found in group A. The positive rate of YMDD mutation was 17.9%, 0, 0, 0 at week 12. There was no significant difference in the level of ALT and the rate of HBeAg seroconversion after 48-week treatment (P > 0.05). At week 48, there was significant difference in the ALT normalization rate and undetectable HBV DNA rate between group C and group A, and also between group D and group A, and the rate of drug resistant genotype was 6.9%, 6.7%, 0, 0. Two patients had rtN236T mutation in group A, and one patient had rtN236T mutation and another one had rtA181V mutation in group B.

CONCLUSIONAdefovir dipivoxil in combination with lamivudine or entecavir are safe and effective therapies for chronic hepatitis B patients with YMDD mutation.

Adenine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Adult ; Alanine Transaminase ; blood ; Antiviral Agents ; administration & dosage ; therapeutic use ; DNA, Viral ; blood ; Drug Resistance, Viral ; Drug Therapy, Combination ; methods ; Female ; Follow-Up Studies ; Guanine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Mutation ; Organophosphonates ; administration & dosage ; therapeutic use ; Reverse Transcriptase Inhibitors ; administration & dosage ; therapeutic use ; Young Adult

We have previously shown that the inhibition of fibroblast growth factor (FGF) signaling induced endodermal gene expression in the animal cap and caused the expansion of the endodermal mass in Xenopus embryos. However, we still do not know whether or not the alteration of FGF signaling controls embryonic cell fate, or when FGF signal blocking is required for endoderm formation in Xenopus. Here, we show that FGF signal blocking in embryonic cells causes their descendants to move into the endodermal region and to express endodermal genes. It is also interesting that blocking FGF signaling between fertilization and embryonic stage 10.5 promotes endoderm formation, but persistent FGF signaling blocking after stage 10.5 restricts endoderm formation and differentiation.
Animals ; Endoderm/drug effects/embryology/*metabolism ; Fibroblast Growth Factors/antagonists & inhibitors/genetics/*physiology ; Gene Expression Regulation, Developmental/drug effects ; In Situ Hybridization ; Pyrroles/administration & dosage/pharmacology ; Receptors, Fibroblast Growth Factor/genetics/physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction/drug effects ; Xenopus Proteins/antagonists & inhibitors/genetics/*physiology ; Xenopus laevis/embryology/genetics/*physiology

OBJECTIVE: To evaluate the long-term efficacy and safety of nevirapine (NVP)-based regimens for HIV-infected Chinese patients in routine clinical practice. METHODS: From October 2002 to May 2004, 57 HIV-1-infected patients commenced highly active antiretroviral therapy (HAART), and were followed to December 2008. They originally received 2 nucleoside reverse transcriptase inhibitors (NRTIs) and nevirapine. HIV RNA levels, T lymphocyte subsets and safety were assessed. Blood routine test and main laboratory parameter changes were traced. If apparent side effects or virological failure appeared we would, if necessary, terminate the therapy or change the regimen. RESULTS: Of the 57 subjects, 34 were followed-up for more than 4 years. After 5-6 years, 63.3% of the subjects (19/30) had HIV RNA levels<50 copies/microL, and the median increase in CD4(+) cell count from the baseline was 329 cells/microL. The mean decrease in CD8(+) cell count was 128 cells/microL. At the same time, the CD4(+) CD45RA+CD62L cell count and CD4(+)CD45RO(+) cell gradually increased, and the counts of CD8(+)CD38(+) cell declined gradually. These changes are apparent 2 years after HAART. The increase rate slowed down after 2 years. But they did not recover completely as well as healthy people at year 6. About 56% (32/57) of HIV-infected patients developed various drug-related side effects. The most common was gastrointestinal side effect, followed nervous disorder, baldness, and rashes, mostly happened in 6 months. Gamma-GT increased occurred in 29.8% of patients (17/57), and serum cholesterol and triglyceride elevated in 26.3% of the patients (15/57). Six patients developed lipodystrophy, mainly in female patients, and 25 patients showed abnormal blood picture and liver function, renal function changes and amylase elevation. Grade 3-4 adverse events occurred in 3 cases (2 peripheral neuropathy, and 1 suspected lactic acidosis). One subject experienced grade 3 rashes. CONCLUSION Antiretroviral therapy with NVP-based regimens is safe and effective by suppressing HIV viremia and producing continued CD4 cell increases in subjects with HIV or AIDS for 6 years.
Adult ; Anti-HIV Agents ; administration & dosage ; adverse effects ; Antiretroviral Therapy, Highly Active ; adverse effects ; methods ; CD4 Lymphocyte Count ; China ; Didanosine ; administration & dosage ; adverse effects ; Female ; Follow-Up Studies ; HIV Infections ; drug therapy ; virology ; HIV-1 ; drug effects ; isolation & purification ; Humans ; Male ; Middle Aged ; Nevirapine ; administration & dosage ; adverse effects ; Reverse Transcriptase Inhibitors ; administration & dosage ; adverse effects ; Stavudine ; administration & dosage ; adverse effects ; Treatment Outcome ; Viral Load ; Young Adult

OBJECTIVETo investigate the pharmacological effects and underlying mechanism of azidothymidine (AZT) on human glioblastoma cells in vitro.

METHODSThe telomerase activity of human glioblastoma TJ905 cells was determined by TRAP assay after 24 hrs' incubation with 50, 100, 200 micromol/L AZT and control vehicle solution. Colony formation efficiencies of the cells were recorded. Cells of the 1st, 3rd and 6th generations were harvested, followed by evaluations of cyclin A protein expression by Western blot, cell cycle distribution by flow cytometry, apoptotic level by single cell gel electrophoresis and proliferation index by Ki-67 immunocytochemical staining.

RESULTSAZT inhibited telomerase activity of TJ905 cells. Cyclin A expression levels in the cells treated with 50 and 100 micromol/L AZT were significantly lower than controls (P < 0.01), and down-regulation of the expression was in a dose- and time-dependent manner. Compared with controls, G(0)/G(1) phase cells were obviously decreased (P < 0.05 approximately 0.01) and S phase cells significantly increased (P < 0.05 approximately 0.01) after treatment with 50, 100 and 200 micromol/L AZT. The cell numbers of G(0)/G(1) and S phases at the 1st generation of above three treated groups changed in a dose-dependent manner, whereas S phase cells increases in all AZT treatment groups and G(0)/G(1) phase cell decrease in group treated with 50 micromol/L AZT were also in a time-dependent manner. Both the apoptotic cells of the 1st and 6th generations of all AZT treatment groups were significantly more than controls (P < 0.05 approximately 0.01), their numbers of the 6th generations of the three groups increased with AZT concentration (P < 0.05 approximately 0.01), and all of them were more than the 1st and 3rd generations of the same dosage group (P < 0.05 approximately 0.01). Colony formation efficiencies and Ki-67 labeling indexes of the three AZT treatment groups were distinctly lower than controls (P < 0.01), and they were also decreased with the elevation of AZT concentration and/or the elongation of the incubating time. The difference of any above parameter had no significance among the 1st, 3rd and 6th generations of control group (P > 0.05).

CONCLUSIONAZT blocks S/G(2) conversion of TJ905 cells by inhibition of telomerase activity and cyclin A expression, leading to an enhancement of apoptosis and suppression of cell proliferation.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin A ; metabolism ; Dose-Response Relationship, Drug ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Glioblastoma ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Reverse Transcriptase Inhibitors ; administration & dosage ; pharmacology ; Telomerase ; metabolism ; Zidovudine ; administration & dosage ; pharmacology

OBJECTIVETo analysis the efficacy and safety of lamivudine (made in China) therapy for 52 weeks in adolescent patients with chronic hepatitis B (CHB).

METHODSOne hundred and five teenage CHB patients were treated with lamivudine 100 mg once daily for 52 weeks. Patients with elevated ALT at baseline were in group 1 and those with normal ALT were in group 2. The changes of HBV DNA, HBV seromarkers and ALT at the end of 12, 24 and 52 weeks after lamivudine therapy were compared with those at baseline. Adverse events were recorded and evaluated.

RESULTSAt the end of 52 weeks of lamivudine therapy, HBV DNA-ve, HBeAg loss and anti-HBe seroconversion were observed in 92.0%, 24.4% and 22.0% in group 1 patients and 76.1%, 14.2% and 14.2% in group 2 patients respectively. No significant differences were found between two groups. At 12, 24 and 52 weeks, normalization rates of ALT were 59.0%, 66.7% and 76.0%, normal ALT with undetectable HBV DNA were 44.9%, 64.1% and 70.7% at the same time. During 52 weeks lamivudine treatment 26 mild adverse events were observed in 18 patients.

CONCLUSIONLamivudine can inhibit HBV replication rapidly and normalize ALT in majority adolescent CHB patients. HBeAg loss or seroconversion of anti-HBe was observed in some of these patients. All patients in this study were safety and well tolerated.

Adolescent ; Alanine Transaminase ; blood ; Antiviral Agents ; adverse effects ; therapeutic use ; Child ; DNA, Viral ; blood ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B e Antigens ; blood ; immunology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Lamivudine ; adverse effects ; therapeutic use ; Male ; Mutation ; genetics ; Reverse Transcriptase Inhibitors ; adverse effects ; therapeutic use ; Safety ; Treatment Outcome