Enzyme-linked immunosorbent assay for determination of thyroid-stimulating antibody in the sera of patients with Graves' disease was established using ganglioside (GLS) purified from bovine brain as binder. It was applied in clinical practice. The results showed that LATS could react with ganglioside under suitable conditions. Very significant difference was observed between normal subjects and untreated GD (P

Objective To explore in vitro effects and the mechanism for FLAG regimen compared with IA regimen in P-glycoprotein-positive and -negative acute myeloblastic leukemia(AML) cell lines. Methods The expression of P-glycoprotein in K562 and K562/A02 cells were analyzed by flow cytometry. The effects of FLAG and IA on the proliferation of K562 and K562/A02 cells were detected by MTT assay. The Ara-CTP and Ara-C levels in those cells were measured by HPLC,the gene expression of hENT1 in K562 and K562/A02 cells was detected by real-time PCR. Results The positive rates of P- glycoprotein were (1.32±0.24)% in K562 cell and (97.66±3.77)% in K562/A02 cell, respectively. The expression of P- glycoprotein had no change after treated with FLAG or IA. The cytotoxicity to K562 of IA was better than FLAG [(84.41±9.33) % v.s (73.17±13.20)%, P<0.05], and the cytotoxicity to K562/A02 of FLAG was better than IA [(70.55±11.32)% v.s (48.46±12.81)%, P<0.01]. Ara-C and Ara-CTP accumulation and hENT1 expression in AML cells treated with FLAG were higher than that treated with IA. Conclusion P-glycoprotein-positive AML cells are more sensitive to FLAG regimen than IA regimen. The biochemical modulation of Ara-CTP and Ara-C may be the major mechanism.

BACKGROUND: Host endothelial cells are important targets of alloreactive donor cytotoxic T lymphocytes in graft-versus-host disease (GVHD). And the expression of tissue factor in vascular endothelial cells is up-regulated post-injury and activation. So tissue factor expression in vascular endothelial cells may play a pivotal role in the pathogenesis of GVHD.OBJECTIVE: To investigate the molecular mechanism of allogeneic T cell-induced tissue factor (TF) expression in vascular endothelial cells.DESIGN, TIME AND SETTING: The cytology observation was performed at immunological laboratory of ongji Medical College, Huazhong University of Science and Technology from March to October 2008.MATERIALS: Human primary umbilical vein endothelial cells (HUVECs) were purchased from Science cell laboratory (CA, USA).METHODS: Allogeneic CD4~+ and CD8~+T cells were isolated from peripheral blood mononuclear cells (PBMC) by positive selection using magnetic beads coated with anti-CD4 or anti-CD8 antibody. After thorough washing with PBS, 1×10~6 allogeneic CD4~+T cells or CD8~+T cells were added to HUVECs (ratio 10:1). The cells were grown at 37 ℃ in a humidified atmosphere containing c-jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated protein kinase (ERK). HUVECs were pretreated with TF antibody 1 hour before co-culturing; the untreated HUVECs were used as controls. MAIN OUTCOME MEASURES: Allogeneic T cells-induced TF expression in vascular endothelial cells was determined by real-time PCR and flow cytometry. Western blot was used to examine allogeneic T cells-induced phosphorylation of MAPK in HUVEC.RESULTS: After co-culturing with allogeneic CD4~+T cells or CD8~+T cells for 6, 12, and 24 hours, the expression rates of TF on HUVEC membrane were obvious increased than that of the control group (P < 0.05), the expression levels of TF mRNA in HUVEC were also significantly elevated (P < 0.05). Allogeneic CD4~+T cells-induced expression rates of TF were significantly higher than allogeneic CD8~+T cells-induced expression rates at 12 and 24 hours after culture. The expression of p38MAPK and JNK was enhanced after culture with allogeneic T cells, yet the ERK expression had no changes. The p38MAPK or JNK can notably down regulated TF expression, which could block the TF expression when worked together. CONCLUSION: Allogeneic T cells induced TF expression in HUVECs via activation of p38MAPK and JNK.

Objective To study the effect of propofol and midazolam on memory during target control infusion for sedation.Methods Forty healthy male volunteers were randomly devided into 4 groups,all subjects received target controlled infusion with propofol or midazolam for sedation.Group 1 and group 3 were sedated to OAA/S 1 or OAA/S 3 respectively by propofol;group 2 and group 4 was sedated to OAA/S 1 or OAA/S 3 by midazolam.All subjects learned a list of words before sedation and at the different sedation depths predicted.After recovery the explicit and implicit memory were tested using word stem completion method and process dissociation procedure.Results Compared with 0,there was no significant difference in explicit memory among all the 4 groups.While there was significant difference in implicit memory for group 3 and 4,and there was no significant difference in implicit memory for group1 and group2.Conclusion Sedative-hypnotic drugs not only deepen sedation,but also impair memory as serum concentration increases during target infusion.Drug-induced amnesia is independent of sedation.Propofol and midazolam have the same efficacy on memory at the same sedation scale.

Objective To explore the expression of protease-aetivated receptor (PAR)-1, 2 in a routine model of acute graft-versus-host disease (aGVHD). Methods A routine model of aGVHD after aUogeneic hematopoietic stem cell transplantation (allo-HSCT) was established, and the syngeneic HSCT mice were used as the controls. Quantitative real-time PCR, Western blot and immunohistoehemistry were done to detect the gene and protein expression of PAR-1, 2 in multiple organs of allo-HSCT mice and the controls. Results Allo-HSCT mice showed classical symptoms and histological changes of aGVHD. PAR-1 mRNA expression was significantly increased in the skin,liver, small intestine of allo-HSCT mice (skin: 0. 039 ± 0. 013 vs. controls: 0. 008 ± 0. 002,P<0. 01 liver: 0. 165 ± 0. 084, vs. controls: 0. 017 ± 0. 006, P<0. 01 ; small intestine: 0. 215 ± 0. 109 vs.controls: 0. 016±0. 009, P<0. 01), but not in the stomach, lung and kidney of allo-HSCT mice (P >0. 05). PAR-2 mRNA expression in the liver and small intestine of allo-HSCT mice was significantly elevated (liver: 0. 010 ± 0. 003 vs. controls: 0. 003 ± 0. 002, P<0. 01 ; small intestine:0. 006 ± 0. 002 vs. controls: 0. 003± 0. 002,P<0. 05), but not in the other organs (P>0. 05). The protein expression of PAR-1, 2 was in accordance with the mRNA expression. Immunohistochemistry revealed the PAR-1, 2 expression was increased in the epithelial eeUs and vascular endothelial cells of target organs of aGVHD. Conclusion Inereased expression of PAR-1, 2 in the target organs of aGVHD suggests PAR-1, 2 may contribute to the pathogenesis of aGVHD after allo-HSCT.

Objective:To analyze that if the quality of ethics review is closely related to the protection of human subjects' right and interest. Methods:This article has analyzed all the issues raised by local Ethics Committee in the process of review in recent two years since guideline of ethical review of drug clinical trials was published, summed up the most common problems occurred in protocols and informed consents. Results:Total 94 new drug or medical device clinical trial projects were reviewed by the local ethics committee, among which 29 projects were ap-proved through regular full board meeting, the approval rate in the initial review was 31%. The most common prob-lems in protocols include: the research backgrounds, design, and risk-benefit ratio; Main issues raised on in-formed consent focused on the contents, language and signature terms. Conclusions:The protection of subjects needs more improvement of capability of investigator, sponsor, drug clinical trial institution and the ethics commit-tee.

Objective: To investigate androgen receptor (AR) mRNA expression in human peripheral leukocytes. Methods: Total RNA was isolated from peripheral leukocytes of healthy men and women, AR mRNA was determined by RT PCR and Northern blot. Results: An AR cDNA fragment of the expected size of 390 bp was determined by RT PCR. 9.4 kb AR mRNA was detected by Northern blot, which was consistent with AR mRNA in human prostate reported by Lubahn et al . Conclusion:AR mRNA expresses in human peripheral leukocytes, this provides basis for studying the effect of androgen on the function of the human peripheral leukocytes and the changes of AR in pathological stages. [

Objective To observe the effectiveness of Antithymocyte globulin/antilymphocyte globulin(ATG/ALG)in the treatment of nonsevere aplastie anemia(NSAA)patients who didn't respond to cyclosporine A(CsA)or relapsed and review the literature.Methods We administered ATG/ALG to 5 NSAA patients who relapsed or showed no re- sponse after CsA therapy.Results 3 patients were cured,2 patients improved,4 patients became transfusion inde- pendent in 90 days,none relapsed or progressed to clonal disease.Conclusion Those NSAA patients who relapsed or showed no response after CsA therapy should be asministered ATG/ALG,so as to reduce transfusion complications and prevent sustained immune attack to the bone marrow.

There are still many problems at liquid state culture especially large-scale culture for Helicobacter pylori at present. Using analytical technique of single factor test statistics, a suitable medium and shaking culture conditions for Helicobacter pylori have been selected. The experimental results showed: under the optimal shaking culture conditions Hp could be growed well enough in Brucella broth with ?-CD. The thalli weight reached 0.6g/100mL.

Objective To develop an HPLC method for simultaneous content determination of ginsenoside Rg1, Re and Rb1 in Panacis Japonici Rhizoma.Methods Chromatographic separation was carried out by using an Agilent Poroshell 120 C18 column (4.6 mm × 100 mm, 2.7μm). The mobile phase consisted of acetonitrile-0.1% phosphoric acid with gradient elution at a flowrate of 1.0 mL/min, and the injection volume was 10μL. The detection wavelength and column temperature were 203 nm and 30℃ respectively.Results Ginsenoside Rg1, Re and Rb1 had the baseline separation and were in good linear range. The recovery rates were 99.5%, 103.0% and 100.5% respectively.Conclusion The approach is simple, accurate, with good repeatability and short analysis period, which can determine the contents of ginsenoside Rg1, Re and Rb1 correctly and provide references for quality control of Panacis Japonici Rhizoma.