Objective To investigate the effect of different stimulating culture conditions and detecting methods on the frequency of Th17 cells and to set up and optimize the experimental method for detecting the frequencies of human Th 17 cells in peripheral blood by flow cytometry (FCM ) .Methods The peripheral blood in the healthy people was performed the stimulating culture by phorbol myristate acetate (PMA) at different final concentrations(50 ,100 ,150 ,200 ,250 ,300 ng/mL) and ionomycin 1 μg/mL .The frequencies of Th17 cells were assessed by FCM after different stimulating time points (4 ,6 ,8 ,10 ,12 h) .The mean fluorescence in-tensity(MFI) and percentage of the ratio of CD4 positive cells on the cellular surface and inside cell membrane were measured be-fore and after PMA stimulation .Results The frequency of Th17 cells was highest with the good stimulation effect under the condi-tion of PMA 150 ng/mL and ionomycin1μg/mL stimulation for 6 h (P<0 .05) .MFI of CD4 inside cellular membrane after stimula-tion was lower than that in the cells with stimulation after labeling membrane surface CD 4(P<0 .05) ,and the Th17 cell frequencies and the ratio of CD4 positive cells were higher than those in the latter ,but the differences were not statistically significant (P>0 .05) .Conclusion After stimulating whole blood with the final concentration of PMA 150ng/mL and ionomycin1μg/mL for 6 h , measuring intracytoplasm CD4 and IL-17 levels is the simple and effective method for detecting the frequency of human Th 17 cells .

Objective To analyze the causes of dual damage during lumbar disc surgery and to probe the prevention and treatment of cerebrospinal fluid leakage after surgery.Methods Clinical materials of 19 cases suffered from cerebrospinal fluid leakage after surgery from March 1997 to October 2001 were retrospectively analyzed.Result Eighteen out of 19 patients were cured by conservative measurements while one was reoperatived for repairing dura mater.Conclusions The rate of cerebrospinal fluid leakage can be decreased by detailed preoperative preparation and strict operation; repairing dura mater in operation and regular conservative treatment after operation can cure most of cerebrospinal fluid leakage. Few cases failing to conservative treatment need reoperation for repairing dura mater.

Objective To investigate the correlation between the expression of human leukocyte antigen B27(HLA-B27)antigen and ankylosing spondylitis(AS),and its implications for the diagnosis and differential diagnosis of AS.Methods Flow cytometry was used to detect HLA-B27.354 suspected AS patients,35 patients final diagnosed with AS and 30 patients diagnosed with rheu-matoid arthritis(RA)were enrolled in the study.Results The positive rate of HLA-B27 in 354 patients with suspected AS was 22.0%,in patients diagnosed with AS was 85.7%,which was higher than the RA group(P <0.05).In AS patients,Males accoun-ted for 77.1%,females accounted for 22.9%,and the patients′age mainly concentrated in youth.The sensitivity and specificity of HLA-B27 test was 85.7% and 100.0% respectively in the diagnosis of AS.Conclusion HLA-B27 antigen detection is important for early diagnosis and differential diagnosis of AS patients.

Adaptive Immunity ; Animals ; Humans ; Periodontitis ; immunology

BACKGROUND:Acute poisoning is frequently encountered at emergency department. This study was to investigate the epidemiology and characteristics of patients with acute poisoning who were treated at the Emergency Center, Fujian Provincial Hospital, China. METHODS:We retrospectively analyzed the gender, age, causes of poisoning, types of poisons, poisoning route, emergency diagnoses, outcomes, and prognoses of these patients. RESULTS:Altogether 2867 patients with acute poisoning were treated from January 2004 to December 2009. The ratio of male to female was 1:1.04, and their average age was 33.8 years. Of the 2867 patients, 76.39％ were between 18 and 40 years old. The incidence of acute poisoning was as high as 11.33％ in January each year. The incidence of poisoning was in a descending order:alcohol poisoning (54.55％), medication poisoning (25.95％), pesticide poisoning (5.65％), and drug poisoning (4.88％). Most (56.44％) of the patients with drug poisoning were under 25 years and their mean age was significantly lower than that of patients with medication poisoning or alcohol poisoning (P < 0.01). Approximately 69.54％ of the patients were followed up after emergency treatment, 30.39％ were hospitalized, and four patients died. CONCLUSIONS:Acute poisoning is largely alcohol poisoning and medication poisoning in a city. The emergency green channel "pre-hospital emergency care-emergency department-hospital treatment"can significantly improve the survival rate of patients with acute poisoning.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; Liposomes ; metabolism ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins c-myc ; metabolism

Exchange Transfusion, Whole Blood ; methods ; Female ; Follow-Up Studies ; Humans ; Hyperbilirubinemia, Neonatal ; blood ; therapy ; Infant, Newborn ; Male

Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans

BACKGROUND: Results from clinical trials suggested that clopidogrel and ticlopidine had side effects of granulopenia, and aspirin could inhibit endothelial progenitor cell proliferation. There is no report of effects of these drugs on human bone marrow mesenchymal stem cells (hBMSCs) in stem cell transplantation. OBJECTIVE: To investigate the effects of antiplatelet drugs including clopidogrel, ticlopidine and aspirin on hBMSC proliferation and secretion. DESIGN, TIME AND SETTING: The cytology in vitro observation was performed at the Laboratory of Toxicology, Shanghai Municipal Center for Disease Control and Prevention from March to December 2006.MATERIALS: The second passage of hBMSCs was kindly donated from Shanghai Tissue Engineering Research & Development Center, Shanghai Ninth People's Hospital. Clopidogrel (Lot number J20040006) and ticlopidine (Lot number H19980186) were obtained from Hangzhou Sanofi-Synthelabo Minsheng Pharmaceutical CO., Ltd. Aspirin (Lot number 20050059) was obtained from Bayer Vital GmbH. METHODS: The standard culture medium consisted of DMEM-LG, 10% heat-inactivated FBS, 100 U/mL penicillin and 100 μg/mL streptomycin. After being cultured in vitro expanded out to passage 6, hBMSCs were treated with antiplatelet drugs of different concentrations and compared with control group. MAIN OUTCOME MEASURES: Cell proliferation was assessed by 3- (4, 5-dimethylthiazol -2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay, level of vascular endothelial growth factor (VEGF) of culture medium was detected by enzyme-linked immunoadsordent assay (ELISA), and surface antigens of hBMSCs were analyzed by the flow cytometry. RESULTS: A570 values of hBMSCs treated by clopidogrel or ticlopidine of 0.02,0.1,0.4,2,10,40 μmol/L were higher than control group (P < 0.01), while A570 values of aspirin group of 60, 600, 2 000 μmol/L were lower than control group(P < 0.05). Antiplatelet drugs had no obvious effect on cell surface antigens(CD34, CD105, CD106)expressed by hBMSCs. Treated by high dose clopidogrel or ticlopidine (40 μmol/L), VEGF level from hMSCs was lower than that of control group(P < 0.01), but VEGF level of low dose (0.02 μmol/L) ticlopidine group was higher than control group(P < 0.01), and there was no significantly difference of VEGF level among low dose clopidogrel group (0.02 μmol/L), aspirin group (5, 2 000 μmol/L), and control group(P > 0.05). CONCLUSION: Clopidogrel and ticlopidine improve proliferation of hBMSCs, but aspirin inhibits proliferation of hBMSCs. High dose of clopidogrel and ticlopidine suppress VEGF secretion of hBMSCs, while low dose of ticlopidine promote it. Antiplatelet drugs have no obvious effect on hBMSCs differentiation.

Objective To investigate the Influence of Survivin and hTERT gene on cell proliferation and apoptosis in human colorectal carcinoma cell line SW 480 and to find experiment evidence for gene therapy of colorectal carci-noma .Methods Plasmids carrying shRNAs targeting survivin and hTERT were designed , constructed and trans-fected into SW480 cells.SW480 cells were then divided into blank group , blank Plasmid control group , survivin RNAi group , hTERT RNAi group and Survivin-hTERT RNAi group .The telomerase activity was examined by TRAP-PCR-ELISA analysis 48h after hTERT-shRNA transfection.Survivin and hTERT mRNA and protein expres-sion was analyzed by RT-PCR and Western blot .Cell apoptosis , proliferation were measured by flow cytometry , CCK-8 assay.Results Telomerase activity of SW480 cells in Survivin-hTERT RNAi groups were significantly decreased compared with the blank group ( P<0.01 ) .The expression of survivin and hTERT mRNA, proteins in the Survivin-hTERT RNAi group was reduced by 82.8%and 73.6%( P<0.01 ) ,79.2%and 66.7%( P<0.01 ) respectively .The inhibitory rate of cell proliferation of Survivin-hTERT RNAi group was 43.6% ±0.1%( P <0.01 ) .The apoptosis rate was 39.2%±2.3%( P<0.01 ) in the Survivin-hTERT RNAi group .Conclusions The Survivin-hTERT RNAi group could significantly reduces the protein expression of survivin and hTERT mRNA, in-hibit cell proliferation and induces cell apoptosis in human colorectal carcinoma cell line SW 480 .