Objective To analyze and discuss how the scanning parameters influence the quality of dual source CT(DSCT) images by the various settings and the corresponding scans.Methods Testees were scanned with corresponding settings of scanning parameters in SOMATOM Definition DSCT produced by SIEMENS and then the results were analyzed and discussed.Results Parameters in relation with the quality of DSCT images could be influenced by the settings of corresponding scanning parameters.Conclusion The settings of scanning parameters have close relationship with the quality of DSCT images,and sometimes one kind of setting can improve image quality in one field but depress image quality in another field,so all factors should be considered synthetically.[Chinese Medical Equipment Journal,2008,29(2):102-104]

OBJECTIVETo establish an appropriate animal model of uterine leiomyoma and to understand the pathogenesis of this disease.

METHODSMature female rats were intramuscularly injected with estradiol benzoate at 200 μg or 300 μg twice a week. After injection for 8 or 10 weeks, the rats were sacrificed. We measured the serum levels of estrogen (E(2)) and progesterone (P), evaluated ER and PR expression, and calculated the leiomyoma forming rate and mortality of the rats. Histological changes were compared between rat uterine leiomyoma and human uterine leiomyoma with HE staining. The optimal dose and duration of E(2) for induction of uterine leiomyoma in rat were determined.

RESULTSIn the rats treated with estradiol benzoate 200 μg for 8 weeks ìn the serum E(2) level increased significantly (P<0.01). Uterine nodules were visible in some of the tested rats. Based on the pathohistological Results , the uterine leiomyoma developed in the treated rats demonstrated similar features as in human uterine leiomyoma. The expressions of ER and PR were increased in the leiomyoma tissues.

CONCLUSIONThe rat model of uterine leiomyoma can be established by intramuscular injection of estradiol benzoate at 200 μg twice per week for 8 weeks, with similar features as those of human uterine leiomyoma. The high concentrations of ER and PR in uterine tissue might be related with the development of uterine leiomyoma in animal.

Animals ; Disease Models, Animal ; Estrogens ; administration & dosage ; adverse effects ; Female ; Leiomyoma ; chemically induced ; Rats ; Uterine Neoplasms ; chemically induced

AIMTo study the stereoselectivity of skin carboxylesterase metabolism and its molecular biological foundation for improving drug percutaneous absorption.

METHODSKetoprofen ethyl ester was used as a model drug, and skin homogenate was applied for studying the stereoselectivity of carboxylesterase metabolism. Human liver L02 cell was used as control of carboxylesterase expression, and RT-PCR was used for studying the expression of carboxylesterase.

RESULTSThe main metabolite of ketoprofen ethyl ester in human skin homogenate was R-ketoprofen. Human carboxylesterase-2 was highly expressed in skin and its cells. However, the expression of human carboxylesterase-1 was very weak or not detectable.

CONCLUSIONHuman carboxylesterase-2 is the main hydrolytic enzyme of prodrugs in percutaneous absorption, and shows metabolic stereoselectivity to prodrugs with chiral esters.

Adult ; Carboxylesterase ; genetics ; metabolism ; Cell Line ; Cells, Cultured ; Humans ; Ketoprofen ; metabolism ; Liver ; cytology ; enzymology ; Prodrugs ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Skin ; enzymology ; Stereoisomerism

Objective To investigate the immunological rejection in the brain of cloning goats received neural stem cell transplantation. Methods Eight cloning goats of CL series were chosen at random and divided into 2 groups. Neural stem cells and saline at the same dosages were transplanted into the fixed site by surgical intervention in the brain cortex of each group, respectively. The levels of IL-2 and IL-10 in the blood of each group were detected at different times (1 w before, and 0, 1 and 3 w,and 3 months after the cell transplantation) to reflect the systemic immune rejection of the goats after the transplatation. The CD3+ cells in the cell transplantation areas in each group were also detected by the method of immunohistochemistry to reflect the local immune rejection after the transplatation. Results The level of IL-2 was obviously higher and the level of IL-10 was obviously lower in the neural stem cell transplantation group than those in the control group 1 and 3 w, and 3 months after the cell transplantation (P＜0.05). The quantity of CD3+ cells in the neural stem cell transplantation group was much larger than that of control groups at the acute period (1 w after cell transplantation) and chronic period (3 months alter cell transplantation, P＜0.05). Conclusion Systemic and local immunological rejections at acute or chronic periods will appear at the brain of cloning goats with neural stem cells transplantation.

AIMTo reconstruct of a tissue engineering skin in vitro for the study of the use of drug percutaneous penetration and metabolism.

METHODSDermal fibroblasts were embedded in collagen type I. HaCaT cells were seeded on the top of the gel. The skin was generated through air-liquid interface culture. Effects of various culture media on tissues morphology were investigated. Sections of the cultured skin were stained with hematoxylin and eosin and examined under microscope. Permeation and metabolism of ketoprofen and its isopropyl ester through the cultured skin were investigated.

RESULTSHaCaT cells initially developed a multilayer epithelium at the air-liquid interface, but it showed a parakeratotic stratum corneum. Vitamin C enhanced cell proliferation obviously. Vitamin D3 promoted cell differentiation. And estradiol showed little effect on the tissue engineering skin. Ketoprofen isopropyl ester was hydrolyzed into ketoprofen when penetrated through the cultured skin, which resembled in the skin cell homogenates metabolism.

CONCLUSIONCultured at the air-liquid interface, HaCaT cells developed a parakeratotic mutilayer epithelium. Enzyme activity was reserved. This cultured skin could serve as an appropriate model for drug percutaneous metabolism and skin irritation.

Administration, Cutaneous ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacokinetics ; Esters ; administration & dosage ; chemistry ; pharmacokinetics ; HeLa Cells ; cytology ; Humans ; Ketoprofen ; administration & dosage ; chemistry ; pharmacokinetics ; Skin Absorption ; Skin, Artificial ; Tissue Engineering ; methods

To investigate the effect of cetirizine hydrochloride on the expression of neurokinin 1 receptor (NK-1R) and cytokines production induced by substance P (SP) in HaCaT cells (a human epidermal keratinocyte cell line) and dermal fibroblasts. The effect of cetirizine on the expression of NK-1R protein was detected by flow cytometry and Western blotting analysis. The modulation of cetirizine on the production of interferon (IFN)-gamma, interleukin (IL)-1beta, IL-6 and IL-8 in HaCaT cells and fibroblasts was measured by ELISA. The results showed that cetirizine significantly inhibited the expression of NK-1R in HaCaT cells and fibroblasts. SP induced the production of IFN-gamma, IL-1beta and IL-8 in both cell types. Cetirizine 1-100 micromol x L(-1) inhibited SP-induced IL-1beta and IL-8 production in HaCaT cells and fibroblasts, while had no effect on the production of IFN-gamma in both cells. Both SP and cetirizine had no effect on the secretion of IL-6 in HaCaT cells and fibroblasts. These findings suggest that cetirizine may be involved in the treatment of SP-induced skin inflammation by inhibiting the expression of substance P receptor and regulation the production of IL-1beta and IL-8 in epidermal keratinocyte and dermal fibroblasts.
Anti-Allergic Agents ; pharmacology ; Cell Line ; Cetirizine ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Histamine H1 Antagonists, Non-Sedating ; pharmacology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-8 ; metabolism ; Keratinocytes ; cytology ; metabolism ; Receptors, Neurokinin-1 ; metabolism ; Substance P ; pharmacology

AIMTo establish an in situ perfused pig ear model for percutaneous absorption.

METHODSThe in situ perfused pig ear model for percutaneous absorption consisted of artificial gas, sample chamber, constant flow pump, constant temperature system, polytetrafluorethylene connective tube, porcine ear vein, porcine ear skin and special laminar flow apparatus. The perfused system viability was assessed by glucose utilization and lactate production. Ketoprofen isopropyl ester and methyl salicylate was used for validating this model. The concentrations of perfused sample were measured by HPLC.

RESULTSGlucose utilization and lactate production showed that this model was viable till 7 h. Ketoprofen isopropyl ester was completely metabolized to ketoprofen in situ in perfused pig ear model. The steady cumulative amount (Q) of ketoprofen from permeation and metabolism was linear with time (t), the equation of ketoprofen formation was Q = -0.024 + 0.120t, the rate of ketoprofen formation was 0.120 microgram.cm-2.h-1. Methyl salicylate was partially metabolized to salicylic acid. The steady cumulative amount (Q) of methyl salicylate from permeation was linear with time (t), the permeation equation of methyl salicylate was Q = -3.809 + 6.129t, the permeation rate of metyl salicylate was 6.129 micrograms.cm-2.h-1. The steady cumulative amount (Q) of salicylic acid from metabolism was also linear with time (t), the formation equation of salicylic acid was Q = -1.785 + 0.879t, the formation rate of salicylic acid was 0.879 microgram.cm-2.h-1.

CONCLUSIONThe in situ pig ear vein perfused model is a novel easy-handing and cost-efficient technique for percutaneous absorption and skin metabolism.

Administration, Cutaneous ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; metabolism ; pharmacokinetics ; Ear, External ; blood supply ; Ketoprofen ; administration & dosage ; metabolism ; pharmacokinetics ; Male ; Models, Animal ; Perfusion ; Salicylates ; administration & dosage ; pharmacokinetics ; Salicylic Acid ; metabolism ; Skin ; metabolism ; Skin Absorption ; Swine ; Veins

In eukaryotes protein phosphorytion is a key event. By reversible protein phosphorylation eukaryotes control many cellular processes including signal transduction, gene expression, the cell cycle etc. Phosphoproteomics involves identification of phosphoproteins and phosphopeptides, localization of the exact residues that are phosphorylated and quantitation of phosphorylation. Because protein phosphorylation is a dynamic process, and it is present at low abundance within cells, and the phosphorylated sites on proteins might vary, and mass spectrometry (MS) signals from phosphopeptides are usually suppressed etc., so phosphoprotein analysis have more difficulties than nonphosphoprotein. In this article, we outline several analysis techniques for separation, identification and quantitation of phosphorylated proteins and peptides, and discuss the progress in these techniques. At present, MS is still an essential core identification technology for phosphoproteomic studies, To search better enrichment strategies are the main challenges in this rapidly evolving field. A major goal of quantitative proteomics is precise quantification and identification of proteins in complex mixtures. A common method for quantitative proteome analysis is the stable isotope labeling method. Today there is no single method that supersedes all others techniques for Phosphoproteomic studies. With continued development of sample preparation techniques and instrumentation, it should be possible to perform a global analysis of protein phosphorylation.
Animals ; Humans ; Mass Spectrometry ; Phosphoproteins ; analysis ; Phosphorylation ; Proteomics ; methods

OBJECTIVESTo explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.

METHODSbcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.

RESULTSbcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.

CONCLUSIONThere are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.

Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Genes, bcl-2 ; genetics ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Polymerase Chain Reaction ; methods

AIMTo investigate the lattice mechanisms involved in the increased dissolution effect of polyethylene glycol (PEG 6,000) dispersion system on poorly soluble drug silymarin (SILY).

METHODSFusion method was used to prepare the solid dispersions of SILY with PEG 6,000. Evaluation of the improvement of dissolution was performed with dissolution studies in vitro. X-ray powder diffraction combined with diffraction peak pattern-fitting procedure were applied to quantitatively analyze the changes of lattice parameters. The interaction of SILY and PEG 6,000 was also determined with Fourier transform-infrared (FT-IR) spectroscopy.

RESULTSThe dissolution rate of SILY was considerably increased when formulated in solid dispersion of PEG 6,000 as compared to pure SILY. The datum from the X-ray diffraction showed the changes in the lattic spacings and particular diffraction peaks (position and the intensity) of PEG 6,000 and SILY. These could explain the increased rate of SILY released from solid dispersion system. The information of FT-IR spectroscopy showed the absence of well-defined drug-polymer interaction.

CONCLUSIONThe dissolution improvement of poorly soluble SILY from solid dispersion of PEG 6,000 can be illuminated by the changes of the lattice parameters of PEG 6,000 and the drug.

Chemistry, Pharmaceutical ; Crystallization ; Crystallography, X-Ray ; Drug Carriers ; Polyethylene Glycols ; chemistry ; Silymarin ; administration & dosage ; chemistry ; Solubility