Animals ; Bacterial Toxins ; administration & dosage ; Biological Transport ; Cell Membrane Permeability ; Drug Delivery Systems ; Endocytosis ; Liposomes ; chemistry ; Peptides ; administration & dosage ; Viral Envelope Proteins ; administration & dosage

The absorption of oral drug in the intestine is an important factor to determine the drug bioavailability. There are many intestinal transporters mediating drug absorption, distribution, excretion and drug-drug interaction. Understanding the transport mechanism can improve the effectiveness and safety of drug and guide clinical rational use of drugs. The in vivo and in vitro methods are used to predict the transport mechanism of drugs by intestinal transporters in the intestine. The purposes of this article are to introduce the main transporters in the intestinal tract, to explain the transport mechanism and to summarize the advantages and disadvantages of the research methods of them.
ATP-Binding Cassette Transporters ; administration & dosage ; metabolism ; Animals ; Anion Transport Proteins ; administration & dosage ; metabolism ; Biological Availability ; Humans ; Intestinal Absorption ; Membrane Transport Proteins ; administration & dosage ; metabolism ; Peptide Transporter 1 ; Symporters ; administration & dosage ; metabolism

The incidence of invasive diseases, including meningitis caused by Haemophilus influenzae type b (Hib) was markedly decreased after routine immunization of Hib vaccine through diverse schedules in many countries. The purpose of this study was to evaluate the immunogenicity and safety of Hib conjugate vaccines in Korean children before the implementation of a national immunization program against Hib in Korea. A multicenter controlled trial was performed on two different Hib vaccines in Korean children. A total of 319 infants were enrolled: 199 infants were immunized with the Hib polysaccharide conjugated to the tetanus toxoid (PRP-T) and 120 infants with the Hib polysaccharide conjugated to the outer-membrane protein of Neisseria meningitides (PRP-OMP). Immunogenicity was evaluated by enzyme-linked immunosorbent assay (ELISA) and serum bactericidal assay. Both vaccines showed good immunologic responses after primary immunization. After 2 doses of PRP-T or PRP-OMP, 78.9% and 91.7% of infants achieved an antibody level of > or = 1.0 microgram/mL, respectively. Both vaccines were safe and well-tolerated. No serious adverse events were observed. Thus, Hib conjugate vaccines appear to be safe and show good immunogenicity in Korean infants. These results will be important reference data for the implementation of Hib vaccine in the national immunization program of Korea.
Bacterial Outer Membrane Proteins/administration & dosage/*adverse ; Enzyme-Linked Immunosorbent Assay ; Haemophilus Vaccines/administration & dosage/*adverse effects/*immunology ; Haemophilus influenzae type b/*immunology ; Humans ; Infant ; Korea ; Polysaccharides, Bacterial/administration & dosage/*adverse effects/*immunology ; Tetanus Toxoid/administration & dosage/*adverse effects/*immunology

OBJECTIVETo investigate therapeutic potential of TRAIL in hepatocellular carcinoma (HCC) and the mechanism of sTRAIL resistance and to reverse the resistance to sTRAIL-inducing apoptosis.

METHODSThe expression profiles of TRAILR were determined 60 HCC samples, in 20 normal liver tissues and 2 HCC cell lines HepG2 and SMMC-7721 by in situ hybridization. Cellular effects of sTRAIL in promoting apoptosis on HCC cell lines HepG2 and SMMC-7721 were analyzed after exposure to recombinant protein and after transfection with a cDNA expression construct. In vivo effects of sTRAIL on tumor growth were investigated using a nude mice HCC model of hepG2. Furthermore, the expression of survivin in HCC was detected, and treatment with antisence oligonucleotide was accepted. Finally, therapeutic effect on HCC by combining sTRAIL and interleukin-12 (IL-12) was detected.

RESULTSBoth DR4 and DR5 were present in all HCC tissues as well as normal hepatic tissues. In contrast, 54 HCC tissues did not express DcR1 and 25 did not express DcR2. But both DcR were detectable in all of the normal liver tissues. The expression patterns of DR and DcR in HCC samples were quite different from those in normal tissue. DR5, DR4, and DcR2 expressed in both cell lines, while no DcR1 expression was detected. Recombinant sTRAIL alone was found to have a slight activity as it killed a maximum of 15% of HCC cells within 24 h while killing over 70% of Jurkat cells. In vivo administration of the TRAIL gene couldn't inhibit tumor growth in a nude mice HCC model. Mostly, HCC tissue and both HCC cell lines expressed survivin, whereas normal liver tissue did not express survivin. Treatment with antisence oligonucleotide enhanced sTRAIL-inducing apoptosis. IL-12 significantly augmented sTRAIL-inducing apoptosis and inhibited survivin expression.

CONCLUSIONSHCC cells are insensitive towards TRAIL-mediated apoptosis. Survivin may play a role in resistance to TRAIL-induced apoptosis in HCC, and antisence oligonucleotide could partly reverse the resistance to TRAIL-inducing apoptosis. IL-12 may sensitize HCC cells to TRAIL-induced apoptosis by preventing survivin. Combining gene therapy strategy such as combining gene therapy of TRAIL with IL-12 may be a promising maneuver to HCC.

Animals ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; Carcinoma, Hepatocellular ; pathology ; therapy ; Cell Line, Tumor ; Genetic Therapy ; Humans ; Inhibitor of Apoptosis Proteins ; Interleukin-12 ; administration & dosage ; genetics ; Liver Neoplasms ; pathology ; therapy ; Membrane Glycoproteins ; administration & dosage ; genetics ; Mice ; Mice, Nude ; Microtubule-Associated Proteins ; antagonists & inhibitors ; Neoplasm Proteins ; Recombinant Proteins ; administration & dosage ; TNF-Related Apoptosis-Inducing Ligand ; Transfection ; Tumor Necrosis Factor-alpha ; administration & dosage ; genetics

OBJECTIVETo detect the immunoreactivity of targeted fusion anti-caries DNA vaccine pGJA-P in vitro, and the ability to enhance the immune responses compared with the non-targeted fusion anti-caries DNA vaccine pGLUA-P.

METHODSThe CHO cells were transfected with pGJA-P and the expression of recombinant protein in cultured supernatants were detected using Western blotting. 5 to 6-month-old female Japanese rabbits were immunized with either pGJA-P or pGLUA-P via either intramuscular injection (i.m.) or intranasal route (i.n.). The sera and saliva were collected and the antibody responses were checked by ELISA. The effect of immune sera on the synthesis of water-insoluble glucan by glucosyltransferase of S. mutans was examined.

RESULTSThe expressed protein could response to specific anti-GTF antibody. The antibody responses in serum generated by pGJA-P via i.m. were significantly higher than those generated by pGLUA-P (P < 0.01). The antibody responses in saliva generated by pGJA-P via i.n. were significantly higher than those generated by pGLUA-P (P < 0.01). The higher mucosal antibody response induced by pGJA-P via i.m. compared with pGLUA-P (P < 0.01) was detected. The immune sera of rabbits immunized by pGJA-P via i.m. most significantly inhibited the synthesis of water-insoluble glucan by glucosyltransferase.

CONCLUSIONSThe recombinant protein expressed by pGJA-P had the immunoreactivity to anti-GTF antibody. pGJA-P could induce faster and higher specific mucosal SIgA antibody responses via i.n. or serum IgG antibody responses via i.m. compared with non-targeted DNA vaccine, pGLUA-P. High titres of specific mucosal antibodies were found in rabbits immunized with pGJA-P via i.m. The immune sera of rabbits immunized by pGJA-P via i.m. displayed the ability of inhibiting the synthesis of water-insoluble glucan by glucosyltransferase.

Animals ; Bacterial Proteins ; immunology ; CHO Cells ; Cricetinae ; Dental Caries ; prevention & control ; Female ; Glucosyltransferases ; immunology ; metabolism ; Immunoglobulin G ; blood ; Membrane Glycoproteins ; immunology ; Rabbits ; Recombinant Fusion Proteins ; administration & dosage ; immunology ; Streptococcus mutans ; immunology ; Transfection ; Vaccines, DNA ; administration & dosage ; immunology

This study was purposed to investigate the effect of bortezomib (Bor) and arsenic trioxide (As(2)O(3)) combination on multiple myeloma cell line KM3 and its mechanisms. KM3 cells were cultured with different concentration of Bor or As(2)O(3) as well as both for a certain time. The cell proliferation was analysed by MTT assay and the concentration of 50% proliferation inhibition (IC(50)) was calculated. Early apoptosis and late apoptosis of KM3 cells were detected by Annexin-V-FITC Kit, and the change of transmembrane potential was measured by flow cytometry. mRNA of Caspase-3, Bim and Bcl-xL were detected by RT-PCR. The results showed that the proliferation inhibitory rate of KM3 cells treated by Bor plus As(2)O(3) was much higher than that of KM3 cells treated by Bor only for 72 h [ (27.64 ± 0.81)% vs (21.67 ± 2.20)%, P < 0.05]. There were more KM3 cells treated by Bor plus As(2)O(3) in early apoptosis at 48 h and late apoptosis at 72 h than that of KM3 cells treated only by Bor [ (53.20 ± 3.70)% vs (35.40 ± 2.58)%, P < 0.01; (63.96 ± 2.97)% vs (54.08 ± 3.76)%, P < 0.01]. Transmembrane potential (Δψm) of KM3 cells treated by Bor plus As(2)O(3) decreased more at 48 h, as compared with Bor alone. The expression levels of caspase-3 mRNA and Bim mRNA in KM3 cells treated with Bor plus As(2)O(3) were higher than that in KM3 cells treated with Bor alone. But the expression level of Bcl-xL mRNA was lower than that in KM3 cells treated with Bor alone. It is concluded that As(2)O(3) can enhance the apoptosis-inducing effect of Bor on multiple myeloma cell line KM3, which is associated with decreasing the expression of Bcl-xl mRNA and increasing the expression of Caspase-3 and Bim mRNA.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Arsenicals ; administration & dosage ; pharmacology ; Bcl-2-Like Protein 11 ; Boronic Acids ; administration & dosage ; pharmacology ; Bortezomib ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Membrane Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Oxides ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins ; metabolism ; Pyrazines ; administration & dosage ; pharmacology ; bcl-X Protein ; metabolism

Relatively little has been studied on the AMA-1 vaccine against Plasmodium vivax and on the plasmid DNA vaccine encoding P. vivax AMA-1 (PvAMA-1). In the present study, a plasmid DNA vaccine encoding AMA-1 of the reemerging Korean P. vivax has been constructed and a preliminary study was done on its cellular immunogenicity to recipient BALB/c mice. The PvAMA-1 gene was cloned and expressed in the plasmid vector UBpcAMA-1, and a protein band of approximately 56.8 kDa was obtained from the transfected COS7 cells. BALB/c mice were immunized intramuscularly or using a gene gun 4 times with the vaccine, and the proportions of splenic T-cell subsets were examined by fluorocytometry at week 2 after the last injection. The spleen cells from intramuscularly injected mice revealed no significant changes in the proportions of CD8+ T-cells and CD4+ T-cells. However, in mice immunized using a gene gun, significantly higher (P<0.05) proportions of CD8+ cells were observed compared to UB vector-injected control mice. The results indicated that cellular immunogenicity of the plasmid DNA vaccine encoding AMA-1 of the reemerging Korean P. vivax was weak when it was injected intramuscularly; however, a promising effect was observed using the gene gun injection technique.
Animals ; Antigens, Protozoan/administration & dosage/genetics/*immunology ; CD8-Positive T-Lymphocytes/*immunology ; COS Cells ; Cercopithecus aethiops ; Humans ; Lymphocyte Activation ; Malaria, Vivax/*immunology/parasitology ; Membrane Proteins/administration & dosage/genetics/*immunology ; Mice ; Mice, Inbred BALB C ; Plasmodium vivax/genetics/*immunology ; Protozoan Proteins/administration & dosage/genetics/*immunology ; Protozoan Vaccines/administration & dosage/genetics/*immunology ; Vaccines, DNA/administration & dosage/genetics/*immunology

OBJECTIVES: Previous studies in patients with Tourette's disorder suggested presynaptic dopaminergic dysfunction, demonstrating increased dopamine densities. In present study, we investigated dopamine transporter densities using I-123N-(3-iodopropen-2-yl)-2beta-carbomethoxy-3beta-(4-chlorophenyl)tropane(I-123-IPT)-SPECT in drug-naive children with Tourette's disorder and postulated that dopamine transporter density reflected dopamine concentrations. METHODS: Eight drug-naive children with Tourette's disorder and six normal children were included in the with the brain SPECT 2 hours after an intravenous administration of I-123-IPT. Obtained SPECT data were reconstructed for the assessment of specific/nonspecific dopamine transporter binding ratio of basal ganglia and were evaluated both quantitatively and qualitatively. We investigated correlation between total tic severity of children with Tourette's disorder assessed with YGTSS and specific/nonspecific binding ratio of basal ganglia. RESULTS: Drug-naive children with Tourette's disorder had a significantly greater increase of speciffic/nonspecific dopamine transporter binding ratio of left basal ganglia than normal children. However, no significant differences in specific/nonspecific dopamine transporter binding ratio of right basal ganglia were found between children with Tourette's disorder and normal children. Also, we found no significant correlation between total tic severity of children with Tourette's disorder and specific/ nonspecific binding ratio of basal ganglia. CONCLUSION: These findings support the hypothesis of dopamine dysregulation in presynaptic dopamine function of the basal ganglia in the pathophysiology of Tourette's disorder.
Administration, Intravenous ; Basal Ganglia* ; Brain ; Child* ; Dopamine Plasma Membrane Transport Proteins* ; Dopamine* ; Humans ; Tics ; Tomography, Emission-Computed, Single-Photon* ; Tourette Syndrome*

OBJECTIVES: It has been suggested that dopamine as well as serotonin were related to the pathophysiology of obsessive-compulsive disorder (OCD). Thus, many studies were performed to nivestigate brain regions and their association with dopamine in OCD patients. Recently, we have been able to monitor the density of the dopamine transporter (DAT) in the basal ganglia using I-123N-(3-iodopropen-2-yl)-2beta-carbomethoxy-3beta-(4-chlorophenyl) tropane (I-123 IPT) SPECT, to evaluate the activity of the presynaptic dopamine function. In present study, we investigated the DAT density of the basal ganglia using I-123 IPT SPECT in patients with OCD. METHODS: Fifteen patients with OCD and nineteen normal control group were included in this study. We performed brain SPECT 2 hours after the intravenous administration of I-123N-(3-iodopropen-2-yl)-2beta-carbomethoxy-3beta-(4-chlorophenyl) tropane (I-123 IPT) and carried out both quantitative and qualitative analyses using the SPECT, which were reconstructed for the assessment of the specific/nonspecific DAT binding ratio in basal ganglia. We then investigated the correlation between the severity of OCD symptoms assessed with the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) and the specific/nonspecific DAT binding ratio of basal ganglia. RESULTS: Patients with OCD showed a significantly increased specific/nonspecific DAT binding ratio in right basal ganglia compared with normal controls and did not show a significantly increased specific/nonspecific DAT binding ratio, and an increased tendency in the specific/nonspecific DAT binding ratio in left basal ganglia (Rt:Z=2.584, P=0.009, Lt:=1.873, P=0.060). We found no significant correlation between the total scores of the Y-BOCS and the specific/nonspecific DAT binding ratio of basal ganglia. CONCLUSIONS: The data of this study suggest that dopamine in basal ganglia plays an important role in fronto-subcortical circuit, which are already known as a site of the pathophysiological mechanism of OCD.
Administration, Intravenous ; Basal Ganglia* ; Brain ; Dopamine Plasma Membrane Transport Proteins* ; Dopamine* ; Humans ; Obsessive-Compulsive Disorder* ; Serotonin ; Tomography, Emission-Computed, Single-Photon*

PURPOSE: Attention deficit hyperactivity disorder (ADHD) has been known as psychiatric disorder in childhood associated with dopamine dysregulation. In present study, we investigated changes in dopamine transporter (DAT) density of the basal ganglias using I-123 N- (3-iodopropen-2-yl) -2-carbomethoxy-3beta- (4-chlorophenyl) tropane [I-123 IPT] SPECT in children with ADHD before and after methylphenidate treatment. MATERIALS AND METHOD: Nine drug-naive children with ADHD and seven normal children were included in the study. We performed brain SPECT two hours after the intravenous administration of I-123 IPT and made both quantitative and qualitative analyses using the obtained SPECT data, which were reconstructed for the assessment of specific/nonspecific DAT binding ratios in the basal ganglia. All children with ADHD reperformed [123I]IPT SPECT after treatment with methylphenidate (0.7mg/kg/d) during about 8 weeks. SPECT data reconstructed for the assessment of specific/nonspecific DAT binding ratio of the basal ganglia were compared between before and after treatment methylphenidate. We investigated correlation between the change of ADHD symptom severity assessed with ADHD rating scale-IV and specific/nonspecific DAT binding ratio of basal ganglia. RESULTS: Children with ADHD had a significantly greater specific/nonspecific DAT binding ratio of the basal ganglia comparing to normal children (Right: z = 2.057, p = 0.041; Left: z = 2.096, p = 0.032). Under treatment with methylphenidate in all children with ADHD, specific/nonspecific DAT binding ratio of both basal ganglia decreased significantly greater than before treatment with methylphenidate (Right: t = 3.239, p = 0.018; Left: t = 3.133, p = 0.020). However, no significant correlation between the change of ADHD symptom severity scores and specific/nonspecific DAT binding ratio of the basal ganglia were found. CONCLUSIONS: These findings support the complex dysregulation of the dopaminergic neurotransmitter system in children with ADHD.
Administration, Intravenous ; Attention Deficit Disorder with Hyperactivity* ; Basal Ganglia* ; Brain ; Child* ; Dopamine Plasma Membrane Transport Proteins* ; Dopamine* ; Humans ; Methylphenidate ; Neurotransmitter Agents ; Tomography, Emission-Computed, Single-Photon