With the development and clinical use of the molecular targeted drugs and individualized treatment,cancer research has been focused on gene targeting diagnosis and treatment.Especially for the epidermal growth factor receptor (EGFR) signaling pathway-related genes,DNA replication-related genes,spindle apparatus format-related genes,cell metabolism-related genes and other molecular targeted detection and treatment,the polymorphism of targeting genes/molecules determines the clinical efficiency of the therapies.

Objective We aim to investigate the association between peptidylarginine deiminase type 4 (PADl4) and colorectal cancer by genotyping threetag single nucleotide polymorphisms (tagSNP) genetic maker, to explore the role of polymorphism of PAD14 gene in development of colorectal cancer. Methods A case-control study was conducted using TaqMan-PCR methods to analyze the genotypes of three TagSNPs such as rs1886302, rs2477131 and rs1635561 for 515 patients with colorectal cancer and 549 health controls. Results There is significantly different in allelic frequencies and genotype frequencies of rs1886302 between cases and controls (P=0.039 and 0.040,respectively). The OR value of A allele and AA genotype is 0.796 and 0.731,respectively,and A allele may reduce the probability of incidence of colorectal cancer. No association Was found among SNPs of rs2477131 and rs1635561 within intronl and intron 15 in terms of coloretal cancer. Conclusion Rs1886302 on PADI4 might be a susceptible SNP to coloretal cancer.

The DNA copy-number variant (CNV) is a kind of segments of DNA ranging from 1 kb to 3 Mb that is present in a variable number of copies.CNVs widely distribute across the human genome,and dramatically increases genetic diversity.In recent years,researches have found that most CNVs are closely related to complex diseases.If a cancer gene is directly encompassed or overlapped by a CNV,it may lead to activation of oncogenes or inactivation of tumor suppressor genes,and finally results in tumorigenesis.CNVs can affect gene expression,phenotype differences and phenotypic adaptations by changing gene dosages and gene activities,and then sequentially lead to tumor or any other genetic dieases.Investigating CNVs is apparently helpful for studing chromosome recombination,genomic evolution,gene expression and the pathogenesis of multiple complex diseases especially tumor.

Objective To investigate the expression of thiredoxin domain containing 5 (TXNDC5) in the synovial tissues and blood samples of various arthritic conditions and autoimmune diseases to further confirm the previous findings, investigate the relations between the expression level of TXNDC5 and clinical parameters of RA. Methods The expression of TXNDC5 in the synovium was quantitatively analyzed by immunohistochemistry, real-time quantitative PCR and Western blotting. The levels of TXNDC5 in blood and synovial fluid was determined using sandwich ELISA in patients with RA, osteoarthritis (OA), systemic lupus erythematosus (SLE), ankylosing spondylitis (AS) and normal controls. One-way ANOVA, LSD test and Spearmen' s correlation were used for statistical analysis. Results Immunohistochemistry indicated that TXNDC5 expression was significantly higher in the synovial tissues of RA (100%, 40±9) than in those of OA and AS(200%,4±4). Real time PCR and western blotting confirmed the above findings (P<0.01). Sandwich-ELISA indicated significantly elevated level of TXNDC5 in the blood and synovial fluid of patients with RA (A=1.31±0.37), but not in those of OA, SLE, and AS, the healthy controls (P<0.01). The level of TXNDC5 in the blood of RA patients (A=0.8185±0.299) was positively correlated with the level of anti-CCP (r=0.350, P =0.027). Conclusion The results suggest that the pronounced increase of TXNDC5 expression may stimulate synovial pannus formation in the hypoxic environment of RA.

The manuscript introduced the overview, training objectives, policy advantages, training process,curriculum, examination of the Australian College in Rural and Remote Medicine and further contrasted that with China's domestics.The authors held that Australia's training is better targeted due to its colleges tailored to this end;the training duration for general practitioners of rural and remote areas is longer,and the training schedule is reasonable;the curriculum design and training content are more targeted;and the homogeneous training is better achieved as its examination is run by the college in a standardized manner.The authors therefore hold that China should develop detailed regulations for general practitioners from rural and remote areas and explore the feasibility of setting up second-level disciplines institutes for internal medicine, surgery, gynecology and obstetrics, pediatrics and general at national and provincial level.

Objective The present study investigated the expression of the citrullinated proteins in the synovium and serum of rheumatoid arthritis(RA)patients.Methods The expression of the citrullinated proteins in the synovium and serum of RA patients was analyzed by two-dimensional western blotting analysis (2-D WB),mass spectrometry MALDI-TOF/TOF MS,western blotting,immunohistochemistry and ELISA.Then we analyzed the data with one-way ANOVA,LSD test,Kruskal-Walls test and Spearman correlation analysis.Results Alpha-1-antitrypsin(A1AT),fibrinogen beta chain(FIBB),keratin type Ⅱ cuticular Hb4(KRT84),tubulin beta chain(TBB)and vimentin(VIME)were detected in RA serum and anti-citrulline antibody could be detected using 2-D WB.A1AT,KRT84 and TBB were expressed significantly in the synovial membranes and synovial fluids of RA patients.Furthermore,high levels of autoantibodies against KRT84 were detected in the blood of RA patients when compared with samples from the healthy controls.Conclusion Current study has identified novel autoantigens in RA,including A1AT,FIBB,KRT84,TBB and VIME using 2-D WB with purified RA sera and anti-citrulline antibody.FIBB and VIME have been confirmed to be autoantigens in the literature,this demonstrates the feasibility of our protocol and the reliability of our study results.

ObjectiveTo investigate the expression of ATA1 in the synovial tissue from patients with ankylosing spondylitis (AS) and to localize expression of ATA1 in AS synovial membranes.In addition,tag SNPs were genotyped to determine the possible association of this gene with AS risk.MethodsWestern blotting analysis was applied to determine the expression of ATA1 in the synovial tissues by comparing the expression profiles of AS(n=8),rheumatoid arthritis(RA,n=9) and osteoarthritis(OA,n=9) samples.Immuno-histochemistry was used to localize the expression of ATA1 in the synovial membrane.The levels of ATA1 in the synovial fluid of patients with AS were determined using ELISA with OA and RA as controls.Taqman method was used to genotype tag SNPs (rs2753934,rs2749531 and rs6575424) in 56 AS cases,260RA cases and 160 healthy controls.ANOVA,LSD test andx2 test were used for statistical analysis.Results Increased expression of ATA1 in synovial membranes of AS was found when compared with samples from RA and OA.ELISA results showed significantly elevated level of ATA1 in the synovial fluid of patients with AS (1.6+0.6),but not in samples of RA(1.4±0.5) and OA (1.2±0.5)(P＜0.05).Haplotype analysis did not reveal a haplotype association in AS or RA(P＞0.05).ConclusionThe current findings suggest that upregulation of ATA1 may play an important role in the pathogenesis of AS.

Objective Carbonic anhydrase 1 (CA1) not only enhances the hydration reaction but also promotes the formation of CaCO3,which is an essential step for new bone formation in vitro.However,there is no direct evidence to demonstrate the involvement of CA1 in bio-mineralization in cells and tissues.This study is aimed to evaluate the important role of CA1 in bio-mineralization and ossification in cultured cells.Methods Calcification in Saos-2 cells was induced using osteogenic medium (OM) and the calcification was determined by Alizarin Red-S staining.The expressions of ossification protein marker Runt-related transcription factor-2 (Runx2),osterix (OSX),alkaline phosphatase (ALP),osteocalcin (OCN) and bone sialoprotein (BSP) were detected in the process of bone formation by real-time PCR.The expression of CA 1 in the calcified cells were measured using real-time PCR and Western blotting.We also detected calcification in Saos-2 cells in the presence of acetazolamide,an anti-carbonic anhydrase drug to CA1,to determine the role of CA1 in biomineralization in culture cells.T test analysis was used to compare the two groups,M-ANOVA of repeated measurs was conducted for different time point.Results Following the stimulation of OM,Saos-2 cells produced a great amount of calcium-rich deposits [0.68±0.03 vs 2.76±0.13,P＜0.01].Increased transcriptions of ossification protein markers were also detected in these stimulated Saos-2 cells,indicating that the OM launched the process of bone formation in the cells.CA1 had a significantly increased expression during this process [0.25±0.03 vs 0.94±0.06,P＜0.01].Following treatment with acetazolamide,the expression of CA1 evidently declined [1.09±0.05 vs 0.55±0.07,P＜0.05],and the mineralized nodule formation was declined [2.76±0.13 vs 2.19±0.07,P＜0.01].Conclusion These findings indicate that CA1 participates in the biomineralization and ossification,and may play an important roles in bone formation.

Objective The present study investigated the expression of heat shock cognate protein 70 (Hsc70) in the synovial tissues and blood samples of patients with rheumatoid arthritis (RA) to determine the pathological role of this protein in the pathogenesis of the disease.Methods The expression of Hsc70 in synovial membranes was quantitatively analyzed by immunohistochemistry,real-time quantitative PCR and western blotting.The samples from osteoarthritis (OA) and ankylosing spondylitis (AS) were used as controls.The levels of Hsc70 in blood of patients with RA were determined using enzyme linked immunosorbent assay (ELISA) with the samples of the healthy subjects as controls.Statistical analysis was conducted with one-way ANOVA,LSD test and Spearmen's correlation.Results Immunohistochemistry showed that Hsc70 had significantly increased expression in synovial tissues of RA than in the samples of OA and AS.Real-time PCR and western blotting confirmed the above findings.ELISA detected significantly elevated level of Hsc70 in blood of patients with RA as compared with samples from the controls (P＜0.01).Conclusion The study suggests that the up-regulation of Hsc70 may be involved in the pathogenic process of RA.

Objective To screen the proteins with decreased expression in the synovial tissues of rheumatoid arthritis (RA) patients by comparing their expression profiles with that of osteoarthritis (OA) and ankylosing spondylitis (AS) patients by a proteomic approach,and to explore the association of reduced expression with disease susceptibility by a single nucleotide polymorphism (SNP) analysis.Methods Proteins extracted from the synovial membranes (n=10 for each disease) were separated by 2-D electrophoresis.The proteins with significantly decreased expression in the RA samples were subjected to MALDI-TOF-MS.The results were verified using Western blotting.Tag SNPs located in the targeted gene were assessed using the Taqman assay in a cohort of 267 Chinese patients with RA and 160 healthy controls.The genotyping results were confirmed in a large cohort of 389 patients with RA and 371 healthy controls.SPSS 11.5 software package was used for one way ANOVA and Fisher's exact test.Results The expression of vitamin D-binding protein (VDBP) in the synovial membranes from patients with RA was significantly decreased when examined by proteomic approach.This result was confirmed by Western blotting analysis.The rs2282679 was significantly associated with RA (P=0.026 794).This result was confirmed in a large cohort of RA( OR=0.678 639,95％CI 0.54l 113-0.851 118,P=0.000 776).Conclusion Compared with samples from patients with OA and AS,RA patients' synovial tissues have low VDBP expression when examined by the proteomic method.The tag SNP rs2282679 located in VDBP is significantly associated with RA.The decreased expression and the genetic effect of VDBP in RA suggest that a novel pathogenic pathway,in which vitamin D contributes,may be involved in the arthritis process of RA.