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

The study was to understand the impact on the proliferation and cytotoxicity of donor's T cells during mobilization with rhG-CSF. The peripheral blood mononuclear cells (PBMNC) were collected from 15 donors before mobilization and on fifth day of mobilization with rhG-CSF. After the PBMNC were activated with 500 ng/ml of CD3 monoclonal antibody and 500 microg/ml of rhIL-2 for 96 hours, the activated T cells were collected for testing proliferation, cytotoxicity, Fas expression, perforin and Fas ligand (FasL) mRNA expression, the IFN-gamma concentration in the culture medium of the activated T cells was determined by radioimmunoassay. The results showed that the proliferation activity of T lymphocytes and the cytotoxicity of T cells activated with CD3 monoclonal antibody and rhIL-2 were reduced markedly after mobilization with rhG-CSF (P < 0.05). The Fas molecule expression in the activated T cells was very high both before and after mobilization with rhG-CSF (P > 0.10). The activated T cells expressed perforin mRNA and didn't express FasL mRNA both before and after mobilization with rhG-CSF. The concentration of IFN-gamma in the culture medium of the activated T cells decreased significantly after mobilization with rhG-CSF (P < 0.01). It is concluded that activity of proliferation and cytotoxicity of donor's T cells is impaired after mobilization with rhG-CSF.
Adolescent ; Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fas Ligand Protein ; biosynthesis ; genetics ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; T-Lymphocytes ; cytology ; drug effects ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; fas Receptor ; biosynthesis ; genetics

OBJECTIVETo establish an porcine model of whole pancreaticoduodenal transplantation with portal venous drainage and enteric drainage for ensuring physiologically normal function without hyperinsulinemia and reducing postoperative complications.

METHODSTwenty sichuan native outbreding white pigs weighing 25-30 kg were divided equally into two groups to serve as the donors and recipients. Cooling of the grafts was accomplished with in situ flush with 4 degrees C UW preservation solution via an aortic cannula. A whole pancreatoduodenal graft with the segment of abdominal aorta and the portal vein was harvested from the donor pigs. Type I diabetes model was established by complete removal of the recipient pancreas. The whole pancreatoduodenal graft was preserved and shaped in UW solution, and the subphrenic abdominal aorta of the recipient was joined with the donor abdominal aorta via a side-to-end anastomosis, and venous reflux was reconstructed between the donor portal vein and the recipient superior mesenteric vein. Side-to-side intestinal anastomosis was performed between the donor duodenum and the recipient jejunum.

RESULTSTen pancreaticoduodenal transplantations (PVE+ED style) were done, and pancreatic graft thrombosis and embolism occurred only in 1 pig 6 days after transplantation.

CONCLUSIONThe model of whole pancreaticoduodenal transplantation with portal venous drainage and enteric drainage is stable and reliable.

Animals ; Drainage ; methods ; Duodenum ; transplantation ; Female ; Intestines ; surgery ; Male ; Models, Animal ; Pancreas Transplantation ; Portal Vein ; surgery ; Swine ; Transplantation, Homologous

The regulation of foreign gene expression in Insect-Baculovirus Expression System is very complex. In this report, the effect of 5'-UTR in the expression of hGH gene in cultured Sf9 cells was examined. A 18 bp length in the end of 5'-UTR of hGH (human Growth Hormone, hGH) cDNA including a stem-loop structure was deleted by PCR. The truncated hGH cDNA, delta 1hGH was cloned in pFastBac1, named pFast-Bac-delta 1hGH. After transforming into E. coli. DH10Bac, which have a shuttle vetor-Bacmid, the delta 1hGH was integrated into Bacmid by site-specific transposition, and an expression vector, rBacmid-delta 1hGH DNA was acquired. By transfecting the cultured Sf9 cells with the recombinant expression vector DNA, pure recombinant virus, rAcV-Bac-delta 1hGH was obtained, and hGH gene was expressed. Immuno-blot and Chemiluminescent assay revealed that the expressed hGH had normal immunological activity, the amount of hGH expression level in Sf9 cell supernatant infected with rAcV-Bac-delta 1hGH containing the truncated 5'UTR was four to five times higher than that infected with rAcV-Bac-hGH.
5' Untranslated Regions ; genetics ; Animals ; Base Sequence ; Cells, Cultured ; DNA, Complementary ; genetics ; Gene Expression Regulation ; Genetic Vectors ; genetics ; Human Growth Hormone ; genetics ; metabolism ; Humans ; Immunoblotting ; Insecta ; cytology ; genetics ; Recombinant Proteins ; isolation & purification ; metabolism