This study is to establish physiologically based pharmacokinetic (PBPK) models of famitinib in rat and monkey, and then to predict the pharmacokinetics and tissue distribution of famitinib in human based on the PBPK models. According to published paper, previous studies and the chemical properties of famitinib predicted by ACD/ADME suite and SimCYP, the PBPK models of rat and monkey were established and optimized using GastroPlus. And then, the PBPK models were applied to predict the pharmacokinetic and tissue distribution of famitinib in human. The results showed that the PBPK models of rat and monkey can fit the observed data well, and the AUC0-∞, ratios of observed and calculated data in rat and monkey were 1.00 and 0.97, respectively. The AUC0-∞, ratios of observed and predicted data in human were 1.63 (rat to human) and 1.57 (monkey to human), respectively. The rat and monkey PBPK models of famitinib were well established, and the PBPK models were applied in predicting pharmacokinetic of famitinib in human successfully. Hence, the PBPK model of famitinib in human could be applied in future drug-drug interaction study.
Animals ; Antineoplastic Agents ; pharmacokinetics ; Haplorhini ; Humans ; Indoles ; pharmacokinetics ; Models, Biological ; Pyrroles ; pharmacokinetics ; Rats ; Receptor Protein-Tyrosine Kinases ; antagonists & inhibitors ; pharmacokinetics ; Tissue Distribution

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

OBJECTIVE: To establish the models of postmortem redistribution(PMR) in dogs with epidural anesthesia and to investigate the effect of temperature on the PMR of Bupivacaine. METHODS: Eighteen male dogs were executed by epidural anesthesia with a dose of 5 mg/kg bupivacaine hydrochloride and randomly divided into three groups, room temperature (20-23 degrees C) group, 4 degrees C group and -20 degrees C group. The cardiac blood, peripheral blood, liver and cerebrum were collected at 0, 2, 4, 8, 24, 48, 72, 96, 120h postmortem. The contents of bupivacaine in those samples were analyzed by GC-NPD and GC-MS, the difference among three groups were compared. RESULTS: The bupivacaine PMR of room temperature group was evident and complex in cardiac blood, peripheral blood and cerebrum. The PMR of 4 degrees C group was weaker and slower than that of normal temperature group. The bupivacaine PMR of the -20 degrees C group was the weakest in three groups. CONCLUSION PMR of bupivacaine will happen in epidural anesthesia death dogs, but it could be delayed or prevent by low temperature storage.
Analgesia, Epidural ; Anesthetics, Local/pharmacokinetics* ; Animals ; Brain/metabolism* ; Bupivacaine/pharmacokinetics* ; Dogs ; Forensic Toxicology ; Gas Chromatography-Mass Spectrometry/methods* ; Liver/metabolism* ; Male ; Models, Animal ; Postmortem Changes ; Temperature ; Time Factors ; Tissue Distribution

Adenocarcinoma ; diagnosis ; Biomarkers, Tumor ; metabolism ; Carcinosarcoma ; diagnosis ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Prostate ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; surgery ; Stromal Cells ; pathology ; Treatment Outcome

Objective To investigate the role of human estrogen receptor-related receptor(ERR) ?,a submember of orphan receptors,in the tumorigenesis of endometrial cancer.Methods Plasmid of pSG-ERR? was transfected into endometrial cancer cell lines HEC-1A,HEC-1B,and Ishikawa.Real-time quantitative RT-PCR and western blot were used to analyze the mRNA and protein expression of ERR? in endometrial cancer cell.Flow cytometry was used to analyze the cellular growth.Results Expressions of the ERR? were significantly increased in the endometrial cancer cells transfected with pSG-ERR? plasmid; expression of the ERR? mRNA in HEC-1A cell was 9644.4 copies/ng,HEC-1B:9835.3 copies/ng,and Ishikawa:8008.6 copies/ng(P

Metabolomics is a new study, which use chromatography, mass spectrometry, nuclear magnetic resonance (NMR), capillary electrophoresis (CE) techniques on the cells, organs and other body fluids and metabolites in samples were isolated, purified and testing, re-use bioinformatics tools on the obtained data are analyzed to obtain one or a set of biomarker information. Based on analysis of the literatures in recent years, metabolomics was summarized from history, concept, advantage, methods, application, difficulties and challenges, journals and books, websites, and its application in forensic medicine was forecasted. As a new branch of global system biology, metabonomics developed rapidly, and its perspective on forensic medicine was feasible and very optimistic.
Biomedical Research/methods* ; Chromatography, Liquid/methods* ; Electronic Data Processing/methods* ; Forensic Medicine/methods* ; Forensic Toxicology/methods* ; Humans ; Magnetic Resonance Spectroscopy ; Mass Spectrometry/methods* ; Metabolomics/trends* ; Pattern Recognition, Automated/methods* ; Specimen Handling/methods*

Based on the records of carboxyhemoglobin in blood samples stored for recent years, the stability of carboxyhemoglobin in these samples could be affected by the containers, the storage temperatures, the volumes of air above the blood, the saturation of the initial carboxyhemoglobin and preservatives added in these blood samples, among which the storage temperatures, the volumes of air above the blood and the saturation of the initial carboxyhemoglobin are the major influence factors.
Air ; Blood Preservation ; Carbon Monoxide/chemistry* ; Carbon Monoxide Poisoning/blood* ; Carboxyhemoglobin/analysis* ; Drug Stability ; Forensic Medicine ; Humans ; Specimen Handling/methods* ; Temperature

Thirst study was purposed to explore the genetic polymorphism of Chinese Zhuang population at HLA-Cw locus by sequence based typing (SBT). A total of 150 unrelated blood samples from Chinese Zhuang population were subjected to sequencing at exon 2, 3 and 4 of HLA-Cw gene in both directions by using SBT technique established by our laboratory. The purified products of sequencing reaction were run by means of electrophoresis on the ABI 3730 DNA Sequencer and the assignment of HLA-Cw genotype was accomplished by using the Assign 3.5 software. The consensus sequence at exon 2, 3 and 4 of HLA-Cw gene for each sample was imported into the Assign 3.5 software. The results showed that 33.33% of tested samples could obtain an unique genotype, genotype in 63.33% of tested samples with ambiguous results could be assigned by ruling out the rare alleles according to the NMDP Rare Allele List File; however, the final genotype in rest 3.33% of the detected samples could be defined when subjected to further confirmatory testing by PCR-SSP. In this detection 16 HLA-Cw alleles were identified, the common alleles with a frequency of > 10% were Cw*0304 > Cw*0102 > Cw*0801 > Cw*0702. The value for gene diversity (GD) was 0.9297, The frequency for Cw*01, 03, 07, 08, 12, 14 (Cw 1 allele group) and Cw*02, 04, 05, 06, 15, 16, 17, 18 (Cw 2 allele group) was 0.8967 and 0.1032, respectively, which indicated that the Cw 1 allele group is the dominant ligand for KIR in Chinese Zhuang population. 51 genotypes were determined and the distribution of genotype frequency was in line with Hardy-Weinberg principle. It is concluded that the obtained HLA-Cw allele frequency and its distribution characteristics of Chinese Zhuang population can provide valuable data in the studies of anthropology and the association of HLA-Cw with disease.
Asian Continental Ancestry Group ; genetics ; Exons ; Gene Frequency ; Genotype ; HLA-C Antigens ; genetics ; Humans ; Molecular Sequence Data ; Polymorphism, Genetic ; Sequence Analysis, DNA

This study was aimed to evaluate whether mesenchymal stem cells (MSCs) obtained from patients with myelodysplastic syndrome possess immunosuppressive effect. MSCs from bone marrow samples of MDS patients were isolated, cultured and expanded. MSCs were morphologically analyzed and their immunophenotype were determined by flow cytometry. Various amounts of MSCs were added into one-way mixed lymphocyte reaction. MSCs from MDS patients were tested for their ability to suppress in vitro proliferation of autologous and allogeneic peripheral blood lymphocytes (PBLs). The results showed that 3 x 10(3 - 1) x 10(5) MSCs from MDS patients could inhibit autologuous PBLs proliferation to (66.9 +/- 20.1)% - (30.2 +/- 5.9)% of maximal response, as well as inhibit allogeneic PBLs proliferation to (56.6 +/- 14.7)% - (20.5% +/- 9.7)% of maximal response, as compared with inhibitory ability of MSCs from healthy donors, there was no significant difference (P>0.05). It is concluded MSCs from patients with myelodysplastic syndrome also possess immunosuppressive activity.
Bone Marrow Cells ; immunology ; pathology ; Cell Proliferation ; Cells, Cultured ; Humans ; Immune Tolerance ; Lymphocyte Culture Test, Mixed ; Lymphocytes ; cytology ; Mesenchymal Stromal Cells ; immunology ; pathology ; Myelodysplastic Syndromes ; immunology ; pathology

The aim of this research was to understand the influence of rhG-CSF on the sphingosine kinase (SphK) activity of monocytes. The peripheral blood monocytes were collected from 6 peripheral blood progenitor cell donors on the fifth day of mobilization with rhG-CSF and from 5 blood donors' buffy coats. The mRNA expressions of monocyte G-CSF receptor and SphK were tested with RT-PCR. The changes of SphK activity of monocytes were assayed after being treated with rhG-CSF. The results showed that the two kinds monocytes collected from both blood donors and peripheral blood progenitor cell donors mobilized with rhG-CSF expressed mRNA of G-CSF receptor and SphK. The SphK activity of monocytes collected from blood donors was not changed significantly after being treated with rhG-CSF (P > 0.05). The SphK activity of monocytes collected from peripheral blood progenitor cell donors transiently increased by (39.6 - 87.2)% after being treated by means of rhG-CSF (P < 0.05) without obviously dose-dependent effect. It is concluded that the SphK activity of monocytes collected from peripheral blood progenitor cell donors can be activated by rhG-CSF.
Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Humans ; Monocytes ; cytology ; enzymology ; Phosphotransferases (Alcohol Group Acceptor) ; drug effects ; metabolism ; Receptors, Granulocyte Colony-Stimulating Factor ; biosynthesis ; genetics ; Recombinant Proteins