Objective: To evaluate the effect of clusterin with and without leader sequence on overexpression preventing apoptosis in human prostate LNCaP cells. Methods:The plasmid pIRES2 EGFP was used to generate the clusterin expression constructs with full length or without the leader sequence (designated as pIRES2 EGFP/cluac, pIRES2 EGFP/clubc, respectively). Western blot analysis was employed to compare clusterin expression levels in the lysis and supernatant fluid of clusterin transfected LNCaP cells in vitro . The distribution of different functional domains of clusterin in cells was detected with Immunocytochemical staining. The clusterin's protective role of Na 2SeO 3 induced apoptosis in LNCaP cells was examined by flow cytometry (FCM) and fluorescence microscope. Results: Clusterin expression was detected in the lysis and supernatant fluid of pIRES2 EGFP/cluac transfected LNCaP cells, while clusterin was found only in lysis liquid of pIRES2 EGFP/clubc transfected LNCaP cells, but not found in their supernatant fluid. The distribution of cluserin in the plasm of pIRES2 EGFP/cluac transfected cells was aggregative, and on the other hand, clusterin distributed dispersedly in pIRES2 EGFP/clubc transfected cells. Its anti apoptotic property in LNCaP cells was proved by FCM and fluorescence microscope.Conclusion: It is apparent that clusterin plays an important role in preventing apoptosis in prostate cancer, and the presence of the leader sequence is necessary for clusterin's anti apoptotic function.

Photoaffinity labeling is widely applied to demonstrate targets of small molecule ligands. In this paper, biotin photoaffinity labeled molecule with propargyl group 1 has been designed and synthesized, followed it's labeling of N2-acetyl-2'-O-propargyl guanosine 9 by "click chemistry". This technology presents delight development potential in labeling of second messenger cyclic nucleotide, antisense oligonucleotide or siRNA.
Biotin ; chemistry ; Click Chemistry ; Guanosine ; chemical synthesis ; chemistry ; Ligands ; Photoaffinity Labels

Photoaffinity labeling is widely applied to demonstrate targets of small molecule ligands. In this paper, biotin photoaffinity labeled molecule with propargyl group 1 has been designed and synthesized, followed it's labeling of N2-acetyl-2'-O-propargyl guanosine 9 by "click chemistry". This technology presents delight development potential in labeling of second messenger cyclic nucleotide, antisense oligonucleotide or siRNA.

Objective:To identify the androgen-responsive genes in prostate and screen the molecular targets for further studying human prostate cancer. Methods:The potential androgen-responsive gene pituitary tumor transforming gene 1 (PTTG1) was selected which had been previously screened by cDNA microarray in rat prostate and its mRNA level was detected by Northern blot in the castrated rat prostate with and without replacement of Mibolerone. Immunohistochemistry was performed to determine the expression and location of PTTG1 in human prostate tissues. Then human androgen-dependent prostate cancer cells LNCaP were used as a model to study the regulation of PTTG1 by Mibolerone. Results: PTTG1 mRNA was hardly detectable in the prostate of 7-day castrated rats, while it was up-regulated dramatically in the prostate of 7-day castrated rats treated with Mibolerone for 2 days. It was showed that high expression of PTTG1 was localized to the epithelial cells of human prostate cancer but not to the stromal cells with Immunohistochemistry. Northern blot analysis indicated that LNCaP cells treated with 0.1 nmol/L Mibolerone for 2 days led to the high PTTG1 mRNA expression. The basic expression of PTTG1 in human androgen-independent prostate cancer cell lines PC3 or DU145 was even higher than that in the human androgen-dependent prostate cancer cells LNCaP treated with Mibolerone. Conclusion: Androgen can up-regulate the PTTG1 expression in castrated rat prostate and human prostate cancer cell LNCaP. It suggests that PTTG1 is potential to play an important role in human prostate cancer progression.

The article reviewed the research on the pharmacological effects of Danshen on hemorheology. We discussed the action mechanism of Danshen and its preparations from the points of hemorheology, blood and vessel endothelium cell. Danshen and its preparations can obviously improve the hemorheological characteristic by various ways.
Animals ; Benzofurans ; isolation & purification ; pharmacology ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hemorheology ; drug effects ; Hemostasis ; drug effects ; Lactates ; isolation & purification ; pharmacology ; Phenanthrenes ; isolation & purification ; pharmacology ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Salvia miltiorrhiza ; chemistry

Clinical practice is a key step that seven-year clinic student take to become doctors.Take the First Affiliated Hospital of Xi'an Jiaotong University as an example,in the medical practice of seven-year clinic students,there still exist some problems such as students' being unable to transform from probationers to doctors,not good at treating with interpersonal relationship in the department and having no perseverant practice attitude.So training before practice should be perfected and supervision should be strengthened to make the seven-year clinic students better.

Objective To understand the information of food hypersensitivity of children with Henoch-Schonlein purpura (HSP) by food intolerance test.Methods Seventy-four children (40 boys and 34 girls;aged from 3 to 14) with HSP who were hospitalized in our hospital from Dec.2006 to May 2007 were chosen.Two mils venous blood was collected from each object,separated serum.Enzymelinked immunosorbent assay method was applied to detect the serum concentration of 14 kinds food allergen IgG in 74 children with HSP.The concentration of food allergen IgG was divided into 4 levels: 0(IgG0.20 U/L,servious allergy).Results The positive rate was 77.0% in 74 cases with HSP.One type of food allergen specific IgG increased in 24 cases(42.1%),2 types of antibodies increased in 16 cases(28.1%),3 types of antibodies increased in 12 cases(21.1%).Four or more types of antibodies increased in 5 cases(8.7%).The positive rates of 14 food allergen IgG were as follow:egg 93.0%,milk 26.3%,soybean 15.8%,tomato 14.0%,wheat 12.3%,ling 12.3%,shrimp 8.8%,crab 8.8%,beef 3.5%,corn 1.8%,rice 1.8%.Conclusion Food intolerance test is a useful method for children with HSP to find food allergen and guideline for diet of these children.

Objective To investigate the protective effect of propofol pretreatment against hepatic ischemia-reperfusion injury and oxidative stress in rats and the mechanism of the role of GSK-3 β.Methods Sixty SD rats were randomly divided into four groups:sham operation group (S group),ischemia-reperfusion group (I-R group),propofol pretreatment group (P group),TDZD-8 pretreatment group (T group).The hepatic ischemia-reperfusion rat models were established by the method of Nauta.Rats were subjected to 30-min,60-min and 90-min 70％ warm ischemia of liver followed by reperfusion for 120 min,respectively.Propofol (12 mg/kg · h) was injected via femoral vein 30 min before ischemia till the end of reperfusion in P group and TDZD-8 (1 mg/kg) were injected via femoral vein 20 min before ischemia in T group.The animals were killed at 120 min after reperfusion.Blood samples and the liver tissue were obtained.The levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),lactate dehydrogenase (LDH),malondialdehyde (MDA) and superoxide dismutase (SOD) were analyzed.Liver morphological changes were observed using optical microscopy.p-GSK-3β Ser9 and total GSK-3 β expression was determined by Western blot.Results Compared with S group,AST,ALT,LDH and MDA level was increased,SOD level was reduced,and p-GSK-3 β Ser9 expression was significantly reduced in I-R group.Compared with I-R group,the content of AST,ALT,LDH and MDA was reduced significantly,SOD increased significantly,and the content of p-GSK-3β Ser9 increased significantly in P group and T group.There were no significant differences between P group and T group.The hematoxylin-eosin staining of hepatic tissues revealed in I-R group had severe structural damage and periportal inflammatory cells infiltrated,hepatocyte necrosis and sinusoidal congestion.In P group and T group,liver tissues had normal structure,less cell death,edema and inflammatory cell infiltration.Conclusions Propofol can significantly reduce hepatic ischemia reperfusion injury by reducing oxidative stress and lipid hydroperoxides.This protective effect of Propofol may be associated with the inhibition of GSK-3 β by GSK-3 β Ser9 phosphorylation.

Polymyxin E shows effective treatment of the infection induced by resistant gramnegative bacteria, but its nephrotoxicity severely limits the clinical application of this drug. In this work, methoxypolyethylene glycols 2000 (mPEG2K)-polymyxin E (PME) was synthesized via chemical grafting reaction and had been characterized. The antimicrobial activity and cytotoxicity of mPEG2K-PME in vitro were investigated on Escherichia coli and HK-2 cells, separately. Intra-abdominal infection model was further established in order to study the therapeutic effect and the toxic effect on kidney of mice. The results showed that mPEG2K-PME exhibited significant inhibitory effect on Escherichia coli and had a lower toxicity on HK-2 cells in vitro. At the same time, mPEG2K-PME had a good efficacy in the treatment of Escherichia coli infected mice in vivo. Moreover, nephrotoxicity caused by mPEG2K-PME was significantly reduced compared to free PME. mPEG2K-PME is promising in development of new preparations with high efficiency and low toxicity.
Animals ; Cell Line ; Colistin ; pharmacology ; toxicity ; Escherichia coli ; drug effects ; Escherichia coli Infections ; drug therapy ; Humans ; Kidney ; cytology ; drug effects ; Mice ; Polyethylene Glycols ; chemistry

This study aimed to explore the impact of depression caused by chronic unpredictable mild stress (CUMS) on in vivo activity of six kinds of CYP450 isoforms in rats. According to 'Katz' method, the model of CUMS was established. Tolbutamide, chlorzoxazone, theophylline, midazolam, omeprazole and dextromethorphan were chosen as probe substrates of CYP2C6, CYP2E1, CYP1A2, CYP3A2, CYP2D1 and CYP2D2 of rats. Plasma concentration of six kinds of CYP450 in control group and model group were determined by LC-MS/MS and computed pharmacokinetic parameters. Consequently, metabolism of theophylline and chlorzoxazone accelerated significantly (P < 0.01), but tolbutamide, dextromethorphan, omeprazole and midazolam had no significant difference. The present study proved that depression caused by CUMS had strong induction to CYP1A2 and medium induction to CYP2E1.
Animals ; Chlorzoxazone ; metabolism ; Chromatography, Liquid ; Cytochrome P-450 Enzyme System ; metabolism ; Depression ; Dextromethorphan ; metabolism ; Liver ; enzymology ; Midazolam ; metabolism ; Omeprazole ; metabolism ; Rats ; Stress, Physiological ; Tandem Mass Spectrometry ; Theophylline ; metabolism ; Tolbutamide ; metabolism