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 objective of this study was to elucidate the correlation of killer immunoglobulin-like receptor (KIR) gene diversity with nasopharyngeal carcinoma (NPC) in the Chinese southern Han population. KIR genotyping of peripheral blood samples from 67 patients with NPC and 77 randomly-selected healthy controls was performed by PCR-SSP, the relative risk (RR) value was calculated by means of Wolf method. The results showed that the KIR2DL3 gene frequency in NPC patient group was significantly lower than that in healthy controls (χ²>3.84, p < 0.05, RR = 0.08), whereas the KIR2DS5 and KIR2DL5B gene frequencies in patient group were significantly higher than those in healthy controls (χ²>3.84, p < 0.05, RR > 1), the other KIR gene frequencies were no statistically different between two groups. It is concluded that the KIR2DL3, KIR2DS5 and KIR2DL5B genes may be correlated with pathogenesis of NPC in the Chinese southern Han population.
Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; Receptors, KIR ; genetics ; Receptors, KIR2DL3 ; genetics ; Receptors, KIR2DL5 ; genetics

OBJECTIVETo explore the effect of burying testis in thigh pocket on spermatogenesis.

METHODSGuizhou miniature male pigs at child-bearing period were randomly divided to receive operation of scrotum incision and dissection with the testis burying in thigh pocket (experimental group) or without (control group). 3 months later, testis biopsy was performed on 2 pigs from each group for pathological examination. Then every male pig from both experimental (n = 6) and control group (n = 6) got a mating partner and lived together for 3 months. The fertility of the male pigs was observed. 6 months after operation, testis biopsy was performed again on all the animals from both the groups.

RESULTSBoth at 3 months and 6 months after operation, the pathological examination showed the spermatogenic cells of all stage in contorted seminiferous tubules markedly decreased with no mature sperm in experimental group, while normal spermatogenic cells with mature sperm in control group. After the male pigs lived with mating partners for 3 months, no female pigs staying with the experimental group became pregnant, but the male pigs in control group had a normal fertility.

CONCLUSIONSBurying testis in thigh pocket impedes spermatogenesis in the miniature male pig. So burying testis in thigh pocket is not recommended for patients with scrotum skin defect who wish to remain fertile.

Animals ; Female ; Fertility ; Male ; Pregnancy ; Pregnancy Rate ; Scrotum ; Skin ; injuries ; Spermatogenesis ; Swine ; Swine, Miniature ; Testis ; physiology ; Thigh ; surgery

OBJECTIVETo explore the expansion method of high purity NK cells from human peripheral blood and explore the changes in biological functions of NK cells after ex vivo expansion.

METHODSNK cells were isolated from peripheral blood mononuclear cells (PBMNCs) by using miniMACS (Magnetic cell-selection) and NK Cell Isolation Kit II, and cultured in SCEM (Stemline Hematopoietic Stem Cell Expansion Medium, Sigma) supplemented with 10% human AB serum and different combinations of interleukin (IL)-2 and/or IL-12, IL-15 for 15 days. Cultures were semi-exchanged with fresh media and cytokines every 3 days. Evaluation for cell expansion, phenotype, perforin and granzyme B mRNA expressions, and IFN-gamma secretion before and after the culture period.

RESULTSCD3(-) CD56(+) cells concentration increased from (11.2 +/- 5.2)% to (94.2 +/- 3.5)%. In group IL-2 + IL-15 and IL-2 + IL-15 + IL-12 group, cells were expanded 50.5 +/- 4.3 and 52.3 +/- 6.7 - fold, respectively, being significantly higher than that in other three groups [(15.4 +/- 1.1 fold in IL-2 group, 19.9 +/- 3.9 fold in IL-2 + IL-12 group, 6.1 +/- 1.0 fold in control group)] (P<0.01), but no significant difference between each other (P>0.05). The purity of CD3(-) CD56(+) NK cells was over 94% in all groups except the control. The perforin and granzyme B mRNA expressions of expanded NK cells in four experimental groups were significantly higher than those of before expansion (P<0.01) and the expressions in IL-2 + IL-15 and in IL-2 + IL-12 + IL-15 group were significant higher than in other three groups (P<0.01) while no significant difference between each other (P>0.05). IFN-gamma levels in the supernatants of four experiment groups were significantly higher than that in control group (P<0.01) and its levels order was IL-2 + IL-15 + IL-12 group > IL-2 + IL-12 group > IL-2 + IL-15 group > IL-2 group (P<0.01).

CONCLUSIONHigh purity NK cells isolated by negative selection using miniMACS can be efficiently expanded with IL-2 + IL-15, and their biological functions were enhanced.

Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Granzymes ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; pharmacology ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Interleukins ; pharmacology ; Killer Cells, Natural ; cytology ; drug effects ; immunology ; metabolism ; Perforin ; metabolism