Objective　Standard method of testing serum fluoride was tested and verified by certified reference material CRM of free fluo ride component in cattle serum (GBW 09143～09144).The method was suited for fluor ide research to assess its practical value.Methods　ISE-standard curve,linear relationship,linear range,sensitive and precision were disired result by CRM.400 serum of fluoride in rats were measured.Results　The linear ranges of the method were 0.02～5.0 mg/L,r=0.9999,b=57.8,and test lower limit was 0.013 mg/L.The test value was equal to standard value.Conclusions　The standard method is accurate,sensitive,convenient and small sample is used;and it is easy to spread.Thus,it has been of practical value.

Anemia, Aplastic ; diagnosis ; etiology ; Benzene ; poisoning ; Expert Testimony ; Humans ; Male ; Occupational Diseases ; diagnosis

Objective To explore the acute toxicity and LD 50 of Tripterygium Hypoglaucum (Levl) Hutch(THH) solution to provide information for safe clinical application. Methods After oral administration of THH solution in mice, the mortality and the physiological and pathological changes were observed. Results The LD 50 (95% confidence limit) of THH in male and female mice was 79 g/kg(69～89 g/kg) and 100 g/kg (90～112 g/kg), respectively. No marked pathological change of the organs was found. Conclusion According to the standard of grading of acute toxicity, THH solution belongs to the moderate class. Therefore, it is safe in clinical practice and has a wide application.

Objective The aim of the study WSfl to establish the method using CDl27 as the new biomarker to identify regulatory T cells(Treg cells)and apply the CD 127 to detect the Treg cells in patients with gastric cancer.Methods The phenotypes of Treg cells were analyzed using five-color flow cytometry method Foxp3.FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7.The mRNA and protein expression of Foxp3 in isolated CD+4 CD25high CD127-/low Treg cells were detected.The relationships between Foxp3 and CD127 protein expression in CD4+ T cells from aduh human peripheral blood were investigated.PBMCs,Ascltes,turnor-infihration lymphocyte and tumor-draining lymph nodes in 35 patients with gastric cancer and PBMCs in 20 normal healthy donors were evaluated for the proportion of Treg ceils,as well as the percentage ot the total CD +4 cells.Results CD4+ CD25 high CD-127 low ceils expressed the high Foxp3 in protein level(87.1%)and mRNA level.Within the CD4+CD+25 population,there was a significant correlation between Foxp3 and the CD127low phenotype(r=0.985,P<0.01).Compared with healthy olunteers,patients with gastric malignancies had a higher proportion of CD4+4 Cdhigh 25 CD-low127 cells in peripheral blood(t=2.542,P<0.05).The Dereentages of Treg cells were more abundant in ascites(t=2.357,P<0.05),TIL(t=6.174,P<0.01) and tumor-draining lymph nodes(t=5.481,P<0.01)of individuals with gastric cancer than that in their blood.There were significant differences in the prevalence of Treg ceils between the early and advanced disease stages in gastric cancer[(6.04±2.31)%in stageⅠ+Ⅱ VB(10.16±2.29)% Ⅲ+Ⅳ,t=2.473,P<0.05].Conclusions The CD127 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies.The populations of CD4+ CD high25 CD127-/low Treg cells increased ith tumor stage in individuals with gastric cancer.

Objective To investigate the populations of CD+4CD25high Foxp3+ regulatory T cells(Treg cells)and mRNA expression of Foxp3 in peripheral and the marginal region of tumor from patients with gastric cancer,and to evaluate the prevalence of Treg cells from patients with gastric cancer in relation to the TNM stages.Methods PBMC,Ascites,tumor-infiltration lymphocyte and tumor-draining lymph nodes in 47 patients with gastric cancer(TNM I11,Ⅱ18,Ⅲ10,Ⅳ8)and PBMC in 20 normal healthy donors were evaluated for the proportion of Treg cells,as a percentage of the total CD4+ cells,by flow cytometric analysis.Levels of mRNA for Foxp3 were measured witll a real-time quantitative PCR Results The percentage of CD4+CD25high Foxp3+ Treg cells in PBMC for cases of gastric cancer were significantly higher than those for healthv donors(P<0.05).The percentage of Treg cells were more abundant in ascites(P< O.05),1rIL(P<0.01)and tumor-draining lymph nodes(P<0.01)of individuals with gastric cancer than in their blood.And the Foxp3 mean fluorescent intensity (MFI) of Treg cells in TIL and tumor draining lymph nodes with gastric caners were significantly higher than PBMC and ascites(P<0.05).The proportion 0f Treg cell in patients of gastric cancers with stage I,II,Ⅲ,Ⅳ were(5.72±1.95)%,(6.45± 2.23)%,(9.58±3.13)%,(11.70±2.30)%,respectively.There were significant correlation between Drevalenee of Treg cells and disease stages in gastric cancer(r=0.784,P<0.01).Foxp3 mRNA levels were higher in PBMC from the group with gastric cancers than in the group with healthy donors.Foxp3 mRNA levels were correlated with TNM stages(r=0.623,P<0.05).Conclusions Our results suggest that the populations of CD4+CDhigh25 Foxp3+Treg cells and Foxp3 mRNA levels in patients with gastric cancer are signifieantlv higher in comparison to those in control donors.In addition,the expression of Fox：p3 mRNA increased with tumor stage.The increased prevalence of Treg cells may be one of the explanations for impaired cell-mediated immunity in cancer bearing hosts.

OBJECTIVETo study interaction between a novel centrosomal protein TACP1 and mitotic kinase Nek2A.

METHODSNek2A305-446 protein was expressed and purified in E.coli and TACP1 protein was expressed in transfected 293T cells. Pull-down assay was used to examine the interaction between Nek2A305-446 and TACP1. TACP1 and Nek2A complex was tested by co-immunoprecipitation assay with polyclonal anti-TACP1 antibody. The localization of those two proteins in Hela cells was verified by immunofluorescence.

RESULTSTACP1 was pulled down by Nek2A305-446 protein but not by GST control. Nek2A was co-precipitated with TACP1 protein by polyclonal anti-TACP1 antibody but not by pre-immunization serum. The Immunofluorescence test showed that these two proteins formed a complex at centrosome during mitosis.

CONCLUSIONCentrosomal protein TACP1 is a novel interacting protein with Nek2A, both of which are localized in centrosome during mitosis.

Cell Line ; Centrosome ; metabolism ; Escherichia coli ; genetics ; Fluorescent Antibody Technique ; HeLa Cells ; Humans ; Immunoprecipitation ; Mitosis ; NIMA-Related Kinases ; Protein Binding ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Telomere-Binding Proteins ; genetics ; metabolism ; Transfection

Objective To investigate the value of ~1H-MRS in the diagnosis and prognosis of diffuse axonal injury(DAI).Methods A prospective imaging study was performed in 63 patients with craniocerebral injury admitted from October 2002 to April 2004.Sixty-three patients were divided into DAI group(27 cases)and Non-DAI group(36 cases)according to the result of the MRI.Then,the ratio of NAA/Cr,Cho/Cr,mINs/Cr,and GIx/Cr at basal ganglia and genu and splenium of corpus callosum was quantified using ~1H-MRS and compared between DAI group and Non-DAI group.Twenty healthy persons were served as control group.The relation between ~1H-MRS indexes and period of primary uneonciousness post-injury was analyzed.Results The results of NAA/Cr and Cho/Cr at genu and splenium of corpus callosum and basal ganglia of control group were 1.19?0.18,1.21?0.24;1.89?0.17,1.84?0.14; 1.57?0.16,1.85?0.25,which of DAI group were 0.83?0.24,2.92?0.78;1.25?0.35,2.54? 0.42;1.33?0.17,2.38?0.44,and those of Non-DAI group were 1.11?0.23,1.61?0.33;1.61? 0.22,1.93?0.26;1.49?0.23,1.89?0.29.The differences between them were statistically significant (P

Objective:To investigate the expression of Bcl-2 and Bax in renal tissues of patients with hepatitis B virus- associated glomerulonephritis(HBV-GN).Methods:Twenty HBV-GN specimens with complete nephrology data and 10 normal renal specimens were randomly chosen for the present study.Cell apoptosis was detected by means of terminal deoxynucleotidyl transferase mediated d-UTP nick end labeling(TUNEL)and the apoptotic index was calculated;immunohistochemistry was used to detect the protein expression of Bax and Bcl-2.ResuLts:The apoptotic index in HBV-GN group was obviously higher than that of the control group;the apoptotic cells were mainly distributed in the proximal and distal renal tubules and the collecting duct epithelial cells,seldom seen in the glomerular cells.The expression of Bcl-2 in HBV-GN patients was predominately present in the renal tubular epithelia cells(positive in the plasma,membrane and nuclear);the expression of Bax was found in both glomerular cells and renal tubular cells,mainly in tubular epithelial cells,seldom seen in Bowman's capsule or glomerular mesangial region.Conclusion:Apoptosis in the kidney of HBV-GN patients mainly occurs in the renal tubular epithelial cells;expression of Bax and Bcl-2 is mainly in the renal tubular epithelial cells,suggesting that the injury of tubular interstitial damage may be one of the important factors for the development of HBV-GN.

OBJECTIVETo discuss the results and significance of the detection of the CFTR gene mutation in azoospermia patients with congenital unilateral absence of the vas deferens (CUAVD).

METHODSWe collected peripheral blood samples from 6 azoospermia patients with CUAVD for detection of the CFTR gene mutations and single nucleotide polymorphisms. We analyzed the genome sequences of the CFTR gene in comparison with the website of the UCSC Genome Browser on Human Dec. 2013 Assembly.

RESULTSMissense mutation of c. 592G > C in exon 6 was found in 1 of the 6 azoospermia patients with CUAVD and splicing mutation of c. 1210-12T[5] was observed in the noncoding region before exon 10 in 2 of the patients, both with the V470 haplotype in exon 11.

CONCLUSIONMutations of the CFTR gene can be detected in azoospermia patients with CUAVD and the detection of the CFTR gene mutation is necessary for these patients.

Azoospermia ; genetics ; Cystic Fibrosis Transmembrane Conductance Regulator ; genetics ; Exons ; Humans ; Male ; Male Urogenital Diseases ; genetics ; Mutation, Missense ; genetics ; Vas Deferens ; abnormalities

Objective To investigate the role of epidermal growth factor receptor and cyclooxygenase-2 pathways in the erlotinib and celecoxib enhanced radiation sensitivity in A549 human lung cancer cell line. Methods IC20 of erlotinib and celecoxibon in A549 human lung cancer cells was measured by MTT assay,Clonogenic assays were used to evaluate the antitumor effects of the drugs and Xirradiation.Flow cytometry was used to assess the apoptosis and cell cycle alteration,and Western blot was used for the detection of Akt and phospho-Akt.Results Both erlotinib and celecoxib could inhibit the proliferation of A549 cells in vitro in a dose-dependent manner and their values of IC20 were (5.15 ± 0.14)and (40.32 ± 1.26) μmol/L,respectively. For radiation survival,the values of Dq,Do,SF2 of the combination of two drugs were lower than those of either drug ( t =6.62,P ＜ 0.05).The SER of celecoxib,erlotinib and their combination were 1.299,1.503 and 2.217,respectively.Flow cytometry assay showed that both celecoxib and erlotinib could enhance radiation-induced G0/G1 arrest,reduce the cell numbmer in S phase,and enhance radiation-induced apoptosis,especially for the combination of drugs.Western blot assay showed that the expressions of Akt protein were similar in all groups.However,pAkt expression was suppresssed by erlotinib and celecoxib,but promoted by radiation.pAkt had the lowest expression in the radiated cells with the treatment of two drugs ( t =4.89,P ＜ 0.05 ).Conclusions The edotinib and/or celecoxib could enhance radiosensitivity probably by increasing cell apoptosis and reducing the number of Sphase cells with low radiosensitivity.