Objective:To investigate the gene polymorphism distribution of tumor necrosis factor-beta(TNF?) with SLE patient and different race in Chinese Han population of Jiangsu province.Methods:DNA samples were extraced from 168 EDTA-blood of unrelated healthy individuals and 66 SLE patients.TNF? alleles were typed using polymerase chain reaction-restrication fragment length polymorphism(PCR-RFLP). Results:The alleles frequencies of TNF? was higher significantly in Chinese Han population than in the White race;the gene frequencies of the TNF?*2 was higher significantly in SLE patients than in the normal controls(SLE patients 67.4%,normal controls 55.1%,P

Objective To investigate the expression and clinical significance of early B-cell factor 3 (EBF3) mRNA and protein in hepatocellular carcinoma(HCC) and the effect of EBF3 overexpression on HepG2 cell proliferation. Methods The expression levels of EBF3 mRNA in 20 pairs surgical specimens of HCC and their distant noncancerous tissues were detected by fluorescence quantitative real-time polymerase chain reaction(FQ-RT-PCR). Western blot was used to detect the expression levels of EBF3 protein in 5 pairs surgical specimens of HCC and distant noncancerous tissues. The fusion protein EBF3-EGFP was ex-pressed in HepG2 cells by transfection of pEGFP/EBF3 into the synchronized HepG2 cells using lipo-fectAMINE 2000 regent. Expression of EBF3-EGFP fusion protein was observed under the inverted fluores-cence microscope. S-phase fraction(SPF) and proliferating index(PI) were analyzed with flow cytometry (FCM). Results The ratio of EBF3 mRNA to β_2 mRNA in HCC tissues was significantly higher than that in distant liver non-cancerous tissues(0.55 ±0.12 versus 0. 22 ± 0.23, t = 5.69, P < 0.001 ). EBF3 protein in nuclear extracts of HCC tissues was about 4 fold that in distant non-cancerous tissues (26.35 ±14.06 versus 7.86 ± 8.47, t = 2.52, P = 0.036). Fluorescence microscopy revealed that 24 h after trans-fection of pEGFP/EBF3 into hepatoma HepG2, the fluorescence of EBF3-EGFP fusion protein was mainly observed in the nucleus. After transfection for 24 h and 48 h, SPF and PI were markedly increased in HepG2 cells transfected by pEGFP/EBF3 as compared with that in pEGFP-N1 transfeeted cells. Conclusion The expression of EBF3 at both mRNA and protein levels was up-regulated in HCC tissues. EBF3 promotes HepG2 cells proliferation through DNA replication, effect of EBF3 in ttCC needs to be further investigated.

Exosomes are small (30 -100 nm) membrane vesicles secreted by various cell types , they mediate cell-to-cell communication by transferring mRNAs , miRNAs, and proteins.As a hotspot in the research of oncology , exosomes have potential values in the research of development , diagnosis and treatment of cancer. This paper briefly reviews the relationship between breast cancer microenvironment , drug resistance and exosomes and its progress in breast cancer diagnosis and treatment , in order to propose new diagnostic and therapeutic strategies .

microRNA (miRNA) is a class of endogenous non-coding small RNAs,they play an important role in post-transcriptional regulation of gene expression through combining the target mRNA that the target protein would not synthesis.The present studies have found that miRNA is involved in many kinds of physiological processes,as well as the pathological processes.The abnormal expression of miRNA in many diseases can be used in diagnosis,prognosis and treatment monitoring.But all of these studies depend on an ideal detection method of miRNA.A lot of detection methods of miRNA expression developed from qualitative analysis to quantitative analysis step by step and from single miRNA detection to high throughput screening in recent years.Various detection methods are improved constantly,the specificity and sensitivity has been improved at the same time,detection procedure become simple and practicable,detection time shorten considerably and cost also reduced ceaselessly,which make the application of miRNA in clinical possible.This review highlights the latest research progress of the common detection methods of miRNA.

Objectives To establish real-time fluorenscence quantitative polymerase chain reaction ( RFQ-PCR ) for measurement of the expression level of guanylyl cyclase-C(GC-C) in the Peripheral blood mononuclear cell(PBMC)in 30 blood donors, 10 cases colorectal cancer tissue and 1 case T84 human colon cancer cell line. Methods Specific primers and TaqMan probe have been designed,and fluorenscence of the PCR product was detected continuously during amplification. According to the standard curve created by plasmid DNA, the expression level of GC-C in clinical samples has been determined using software, and the results were presented as the ratios of GC-C mRNA to?2-microgluobulin(?2M)mRNA. Results The detection range of the assay was from 101 pg/ml to109pg/ml,the coefficient of variation values of both intra-experimental and inter- experimental reproducibility were 6. 87% to 11. 12% and 8. 86% to 15. 19% . None of 30 blood donors and 11 benign intestinal patients expressed GC-C mRNA,it was expressed in 31/37 colorectal cancer patients. The expression level of GC-C mRNA in colorectal cancer patients was 0. 88?0.06,and the expression level of its in colorectal carcinoma tissue and T84 cells were 0. 86?0.07/ug tissue and 0.0082/per cell. Conclusions This assay had high sensitivity,specificity and reproducibility.

Objective To investigate the existence and variance of quinolone-resistance genes in a group of pan-drug resistant of Acinetobacter baumannii ( A.baumannii ).Methods Twenty strains of pandrug resistant A.baumannii were isolated from patients registered in Affiliated Hospital of Nantong University from January 2011 to April 2011.Drug target genes to quinolone (gyrA,parC) and quinolone-resistance genes mediated by mobile genetic elements [ qnrA,qnrB,qnrS,aac(6')-Ⅰ b-cr,qepA] were analyzed by PCR and verified by DNA sequencing.Results In all 20 strains of A.baumannii,the sense mutation was found in the quinolone resistance-determining region of the gyrA gene in the form of TCA to TTA at codon 83 (Ser-83-Leu).Moreover,in the quinolone resistance-determining region of the parC gene sense mutation was found in the form of TCG to TTG at codon 80 (Ser-80-Leu) and 3 synonymous mutations were CCC to CCT at codon 40,GTA to GT]T at codon 41 and CGT to CGC at codon 44.And parC gene was a new mutation.However,mutations were not found in quinolone-resistance genes mediated by mobile genetic elements [ qnrA,qnrB,qnrS,aac( 6 ' )-Ⅰ b-cr,qepA ].Conclusion Quinolone-resistance-determining region play a key role in resistance to quinolones in this group of A.baumannii.To our knowledge,this is first report about the emergence of the new mutation of parC gene in China.

Objective:To clone the encoding sequence of human EBF3 gene,construct recombinant eukaryotic expression plasmid vector pEGFP/EBF3,and study the effect of EBF3 on HepG2 cell cycling.Methods:Total RNA was isolated from placental tissue.Full-length human EBF3 cDNA was amplified by RT-PCR,cloned into eukaryotic expression plasmid vector pEGFP-N1 and sequenced.The expression and sub-cellular localization of the fusion protein EBF3-EGFP in HepG2 cells were analyzed by Western blot.Cell cycles were analyzed with flow cytometry analysis.Results:Obtained full encoding sequence of early B cell factor 3 was identical with that included in GeneBank,and the eukaryotic expression plasmid vector pEGFP/EBF3 was constructed correctly.24 h after transfected by pEGFP/EBF3,the fusion protein EBF3-EGFP was observed mainly in the cellular nucleus under the inverted fluorescence microscope.Western blot analysis confirmed that the EBF3-EGFP fusion proteins of Mr 87 000 were detected in both cytoplasmic and nuclear protein of the HepG2 transfected by pEGFP/EBF3 for 24 h or 48 h.Flow cytometry analysis revealed that the percentage of cells in the S phase was markedly increased in HepG2 cells transfected by pEGFP/EBF3 as compared with that in pEGFP-N1 transfected cells.These findings suggested that transfection of EBF3 gene into HepG2 induced cell proliferation by increasing the number of cells from G1 phase to G2 phase.Conclusion:The recombinant eukaryotic expression plasmid vector pEGFP/EBF3 is successfully established.The percentage of cells in the S phase is markedly increased in pEGFP/EBF3 transfected cells as opposed to pEGFP-N1 transfected cells.It is likely that EBF3 promotes HepG2 cells proliferation through DNA replication.

Objective To study the influence of short hairpin RNA-mediated inhibition of APRIL gene on proliferation,invasion and metastasis of human colon carcinoma SW480 ceils in vitro. Methods After treatment with shRNA, the expression level of APRIL protein in human colon carcinoma SW480 cells was detected by western blotting. And cell adhesion, cell migration and cell invasion of SW480 cells transfected with sh637 were analyzed respectively as compared with non-transfected SW480 cells and mocktransfectant. Results The expression of APRIL protein in SW480 transfected with sh637 were significantly lower than mock-plasmid groups and untransfected ones (F=42.15, P=0.00). Cell proliferation was markedly inhibited, compared with untransfected groups and negative control ones (F=24.76, P<0.05).And the average absorption of cell adhesion in transfected groups, mock-plasmid and non-transfected ones were 0.343, 0.409, 0.412, respectively. Cell adhesion decreased 39% compared with untransfected groups. Similarly, there was significant difference for cell migration (F=65.53, P<0.01) between transfected groups (The average ceil number in exposed area to migrate was 47.89±13.16) and nontransfected (98.78±23.26) as well as mock-plasmid ones (108.01±39.11). Then, in view of the average absorption of cell invasion between transfected groups(0.58±0.10) and non-transfected (0.94±0.23) as well as mock-plasmid ones(1.11±0.21), a prominently difference for cell invasion was also found between them (F=5.771, P<0.05). Conclusion The results suggest APRIL has a proliferation-inducing effect and probably be involved in cell invasion and cell metastasis of colon carcinoma and it could enhance capacity of invasion and metastasis of SW480 cells.

Long non-coding RNAs are a novel class of non-protein coding RNA molecules with more than 200 nucleotides in length.Recently, mounting evidence has been demonstrated that lncRNAs play crucial roles in epigenetic modification and gene expression regulation. This review aims to summarize current knowledge of lncRNAs in the carcinogenesis, clinical diagnosis, drug resistance and prognosis prediction of gastric cancer.

Recent studies demonstrate that the long non-coding RNAs (lncRNAs), pseudogenes, circular RNAs (circRNAs) and so on can be used as competitive endogenous RNAs (ceRNAs), binding to microRNAs (miRNAs), to regulate the expression level of their targets.This novel interplay optimizes traditional liner miRNA→RNA pattern and has become a research hotspot in the scientific community.Dysregulation of ceRNA interplay will influence the body′s normal life activities, leading to the occurrence of diseases, and even tumor formation.This article briefly introduces the key components of ceRNA crosstalk and its research progress in gastric cancer pathogenesis, so as to provide a new thought for cancer research, clinical diagnosis and therapeutics.