Objective To explore the association between blood cadmium levels (BCLs) and clinicopathological characteristics of patients with breast cancer.Methods The clinicopathological characteristics and blood specimens of 186 patients diagnosed with breast cancer were collected between July and December 2009.BCLs were detected by graphite-furnace atomizer absorption spectrophotometer.Mann-Whitney U test and Kruskal-Wallis H test were used to compare the BCLs of patients with different clinical characteristics.Spearman rank correlation analysis was used to evaluate the relationships between BCLs and some indices of clinical characteristics.Results The BCLs was 2.280 (1.579) μg/L.The BCLs were significantly different in patients with different age and body mass index (BMI) (Z =-2.075,P =0.038;x2 =7.429,P =0.023).Also,there were significant differences in different T stages (x2 =10.137,P =0.017),M stages (Z =-2.225,P =0.026),clinical stages (x2 =16.060,P =0.001) and human epidermal growth factor receptor-2 (HER-2) status (Z=-2.072,P=0.038).Excepting for age (r =0.126,P =0.066),BCLs were positively associated with BMI,T stage,M stage,clinical stage and HER-2 status (r=0.159,P =0.030;r =0.171,P=0.020;r =0.166,P =0.044;r=0.154,P =0.040;r =0.152,P =0.038).Conclusion The BCLs are associated with some clinicopathological characteristics of breast cancer,and high blood cadmium burden may promote the development of breast cancer.

Objective To predict the epitopes of Mycobacterium tuberculosis Rv2657c protein, to understand its immunogenicity Methods The T?cell and B?cell epitopes of Mycobacterium tuberculosis Rv2657c protein were predicted by DNAStar software package. The homology of Rv2657c amino acid sequence with the human protein sequences was prepared using Blast method, then the CTL epitopes were predicted using SYFPEITHI supermotif method, BIMAS quantitative motif method and NetCTL prediction method, and the Th epitopes were predicted by RANKPEP and SYFPEITHI supermotif prediction method. Results The prediction using DNAStar software package showed that Rv2657c protein had 5 B?cell epitopes and 6 T?cell epitopes. The protein had 6 CTL epitopes and there were 38 Th epitopes. Conclusion Rv2657c protein has both B?cell epitopes and T?cell epitopes. It may be a candidate target antigen for the studies of vaccine and diagnosis of tuberculosis.

Objective To investigate the Mycobacterium tuberculosis infections in 2014 among Beijing army recruits, and evaluate a new method for screening latent tuberculosis infections.Methods A total of 194 army recruits were subjected to chest X-ray examination purified protein derivative(PPD) skin test, antibody detection, and interferon gamma release assay(IGRA) by ELISA combined with recombinant protein CFP10-ESAT6 and latent infection protein Rv2628.Results The positive rates of PPD skin test and antibody test were 49.7% and 15.5%, respectively.The latent infection rate of IGRA test was 22.2% in 194 cases after CFP10-ESAT6 stimulation.After stimulation of latent tuberculosis infection(LTBI) with Rv2628, IFN-γ level was significantly higher than that in healthy control group (P<0.05).The weak positive group of TST (5 mm≤diameter<15 mm) had a significantly higher level of IFN-γ than the strong positive group(diameter≥15 mm)(P<0.05),but after stimulation with CFP10-ESAT6,IFN-γ levels were not significantly different between the two groups(P>0.05).There was no significant difference between antibody negative and positive groups after stimulation by CFP10-ESAT6 and Rv2628 (P>0.05).The area under the ROC curve of Rv2628 diagnosis of tuberculosis infection was 0.84.When Youden index was 0.621,the specificity was 94.7% and sensitivity was 67.4%.ConclusionCombined detection of antigens Rv2628 and CFP10-ESAT6 specific IFN-γ values can be potentially used for differential diagnosis of active or latent tuberculosis infections.

Objective:To summarize the research advance in antitumor activity of dandelion in recent 5 years.Methods:The lit-eratures on the antitumor activity of various extracts of dandelion in recent 5 years were collected , including the effective components such as polysaccharide , triterpenes , phytosterols , flavonoids and organic acids etc .Results:Dandelion showed antitumor activity .The polysaccharide ingredients could effectively prevent the proliferation of pancreatic cancer PANC -1 cells, lupeol could reduce the activity of breast cancer MCF-7 cells, and beta sitosterol could induce the apoptosis of human cervical cancer Hela cells .Conclusion:Dande-lion is effective in the treatment of pancreatic cancer , breast cancer and cervical cance etc .The antitumor activity of the other active in-gredients in dandelion needs further research in order to provide complete theoretical and experimental evidence for the potential appli -cation as antitumor medicine in clinics .

Objective To investigate the distribution of 3279 and 924 Foxp3 genotypes in Guangdong population and to explore the correlation between Foxp3 gene polymorphism and essential hypertention . Methods Two hundred and six essential hypertention patients and 291 healthycontrols from October 2013 to September 2014 in the third affiliated hospital of guangzhou medical university were enrolled in the study. The Foxp3 3279 and 924 genotypes was identified by PCR-SSP assay. The plasma lipid level and other risk factor were detemined in all subjects. The relationship between genotypes and pathogenesis of EH was analyzed. Results There were sigificant differences in frequecncies of allele and genotype distribution in Foxp3 3279 genotypes between the two groups. The frequecies of AC+CC and allele were significantly higher in the EH group than those in the control group (P<0.05). Result of logictics analysis showed that AC+CC genotype significantly increase the risk of EH (OR = 1.552,95%CI为1.021 ~ 2.357, P < 0.05), but the polymorphism of Foxp3 924 genotype frequecncy was not assosiated with EH. Conclusion The Foxp3 3279 gene polymorphisms is associated with EH . However, the Foxp3 924 gene polymorphisms is not associated with EH.

AIM: To investigate the regulatory function of bone marrow-derived mesenchymal stem cells (MSCs) on T helper 17 cells (Th17) and regulatory T cells (Treg) in peripheral blood of severe asthmatic children . METHODS:MSCs were isolated , cultured and identified in vitro.MSCs digested with mitomycin were cocultured with T lymphocytes (TLC) at different ratios (1∶1, 1∶2, 1∶10 and 1∶20) from severe asthmatic children for 72 h.The prolifera-tion of TLC was measured by CCK-8 method.In the coculture system of the 1∶2 ratio and the single TLC system , the super-natant levels of interleukin-17 (IL-17) and transforming growth factor-β(TGF-β) were measured by ELISA.The mRNA expression of retinoic acid-related orphan nuclear receptor C (RORC) and forkhead box protein 3 (Foxp3) in TLC was de-tected by qRT-PCR.RESULTS:After cocultured with MSCs , the proliferation of TLC decreased significantly in a dose-dependent manner (P<0.05).It also showed decreases in IL-17 (3 799 ±441 vs 4 890 ±373, P<0.05) and RORC mRNA level (1.21 ±0.14 vs 3.85 ±0.48, P<0.05), while an increase in TGF-βlevel (209 ±32 vs 117 ±26, P<0.05) was observed.No influence on the mRNA expression of Foxp3 was found (P>0.05).CONCLUSION: MSCs suppresses Th17 polarization of naive peripheral blood CD 4 +T cells and matures Th17 cells secreting IL-17, which may ef-fectively revise Th17/Treg imbalance of asthma .

Objective To investigate the expression of aquaporins-3 (AQP3) in amniotic epithelial cells regulated by cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signal pathway and to explore the mechanisms of its expression.Methods The amniotic epithelial cells were collected from 30 patients who underwent elective caesarean sections at term with normal amniotic fluid volume and primarily cultured.The cultured cells were treated with (1) forskolin groups: different concentration (0,2.5,5,50 or 100 μmol/L) of forskolin treated cells for 2 hours,and the optimal concentration of forskolin treated cells with different time (0,1,2,10 or 20 hours) ; (2)SP-cAMP groups: different concentration (0,2.5,5,50 or 100 μmol/L) of SP-cAMP treated cells for 2 hours,and the optimal concentration of SP-cAMP treated cells with different time (0,1,2,10 or 20 hours); (3)H-89 groups: different concentration (0,5,10,50 or 100 μmol/L) of H-89 treated cells for 2 hours,and the optimal concentration of H-89 treated cells with different time (0,1,2,10 or 20 hours).The level of intracellular cAMP and activity of PKA were detected by using ELISA,and immunohistochemistry was used to detect the localization of AQP3,the protein expression of total cAMP-response element binding protein (CREB) and phospho-CREB (p-CREB) and AQP3 were assessed by western blot analysis.Cell proliferation was assessed by cell counting kit-8 (CCK-8)assay.Results (1) The brown staining of AQP3 was detected in both cell membrane and cytoplasm in each group.(2) There was no significant change of the cell proliferation rate among groups with different concentration of forskolin,SP-cAMP and H-89 treatment (P ＞ 0.05).(3) After different concentration of forskolin treated 2 hours,the expression of total CREB had no significant difference among them(P ＞ 0.05).While the expression of cAMP level,PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 2.5 μmol/L,5 μmol/L,50 μmol/L forskolin group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 5 μmol/L forskolin group were higher than that in 2.5 μmol/L and 50 μmol/L (P ＜ 0.05).The optimal forskolin concentration was 5 μmol/L.(4) After different concentration of SP-cAMP treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 5 μμmol/L,50 μmol/L SP-cAMP group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 50 μmol/L SP-cAMP group were higher than that in 5 μmol/L (P ＜0.05).The optimal SP-cAMP concentration was 50 μmol/L (5) After different concentration of H-89 treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were lower in 10 μmol/L,50 μmol/L and 100 μmol/L H-89 group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 10 μmol/L H-89 group were lower than that in 50 μmol/L,100 μmol/L (P ＜ 0.05).The optimal H-89 concentration was 10 μmol/L.(6) p-CREB and AQP3 protein expression were significantly lower in 5 μmol/L forskolin combined 10 μmol/L H-89 incubating 2 hours group when compared with 5 μmol/L forskolin,but higher than that in 10 μmol/L H-89 treated group (P ＜ 0.05).Total CREB was no significant difference among the three groups (P ＞ 0.05).Conclusion cAMP-PKA signal transduction pathway may regulate AQP3 protein expression in human amniotic epithelial cells.

Objective: To identify Xanthii Fructus and secure its quality and safety in medication. Methods: Total ge-nomic DNA was extracted from Xanthii Fructus and its adulterants. ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner V 4.2. The Kimura 2-Parameter (K2P) distances were calculated using MEGA 5.0. The neigh-bor-joining (NJ) phylogenetic trees were constructed. Results: The intraspecific genetic distances of Xanthii Fructus were 0. The interspecific genetic distances between Xanthii Fructus and its adulterants were ranged from 0.009 to 0.542. The NJ tree showed that Xanthii Fructus could differ from its adulterants obviously. Conclusion: ITS2 can be used to identify Xanthii Fructus from its adulterants effectively, and our study further confirmed the effectiveness of ITS2 to identify traditional Chinese medicinal materials.

Objective To construct recombinant plasmid containing CFP-10 gene of Mycobacterium tuberculosis(MTB).Methods The gene fragment of CFP-10 was amplified by PCR from Mycobacterium tuberculosis H37Rv genomic DNA and cloned to pET-32a(+) vector.The recombinant plasmid pET-32a-CFP-10 was transformed into E.coli BL21 (DE3) and induced by IPTG.Results CFP-10 gene fragment was amplified from genomic DNA of Mycobacterium tuberculosis H37Rv strain,and thepET-32a(+) prokaryotic recombinant plasmid was constructed successfully.The recombinant protein was expressed with the induction of IPTG.Conclusions The prokaryotic expression vector for CFP-10 was successfully constructed and the recombinant protein was highly expressed in E.coli BL21 (DE3),which lays a foundation for its subsequent immunological function study.

Objective To study the correlation between genetic polymorphisms of interleukin (IL)-1A/1B and susceptibility to tuberculosis (TB).Methods Genetic polymorphisms of IL-1A and IL1 B in 1032 TB patients and 1008 non-TB patients were analyzed using PCR-MassARRAY method.The correlation between genetic polymorphisms of IL-1A/1B and susceptibility to TB was statistically analyzed.Results Two tag SNPs of IL-1A and three tag SNPs of IL-1B were screened for the study.There were differences in the allele frequencies of rs2853550 and rs3783526 between TB group and non-TB group (P=0.047and P =0.034,respectively).IL-1 B SNP1 rs2853550 (P =0.025,OR =1.302,95 ％ CI =1.034-1.640,TC vs.CC) was found to be highly associated with TB,while the other SNPs showed no significant correlations with TB.Furthermore,IL-1B SNP1 rs2853550 [P=0.019,OR=1.308,95％ CI=1.045-1.638 for (TC+TT) vs.CC] in the dominant model conferred significant risk for TB,but IL-1A SNP2 rs3783526 [P=0.000,OR=0.764,95％ CI =0.591-0.988 for GG vs.(AA+GA)] in the recessive model showed protective effects against TB.The haplotype ‘TG’ in the IL-1B block showed a higher risk for TB compared with the common ‘ CA’ haplotype (P=0.032,OR=1.265,95％ CI=1.020-1.567).The diplotypes containing ‘ GA’ haplotype in IL-1A block and ‘ TG’ haplotype in IL-1B block were major risk factors for TB (for onecopy,adjusted P=0.014,OR=1.403 and 95％ CI=1.072-1.836; adjusted P=0.013,OR=1.339 and 95％ CI=1.063-1.688,respectively),but the diplotype with ‘CG’ in IL-1B block played a protective effect against TB (for two-copy,P=0.006,OR=0.664 and95％ CI=0.494-0.891).Conclusion The genetic polymorphisms of IL-1B rs2853550 might be closely associated with TB,but the GG genotype of IL1 A SNP rs3783526 might have the characteristic of anti-TB.