AIM: To explore the relationship between methylation status of promoter region 5′CpG island and the biological phenotype in human colorectal cancer RKO cell lines. METHODS: RKO cells were treated with selective DNA methyltransferase (DNMTs) inhibitor, 5-Aza-2′-deoxycytidine (5-Aza-CdR), for 72 h. Methylation-specific PCR (MSP), T-A clone and DNA sequence analysis were used to detect 5′CpG island methylation status of p16/CDKN2 tumor suppresor gene. Cell growth, cell cycle arrest and apoptosis were analyzed by MTT, flow cytometry (FCM), fluorescent dye staining and transmission electron microscope. RESULTS: DNMTs inhibitor (5-Aza-CdR) effectively reversed the hypermethylation status of 5′CpG island. The effects of 5-Aza-CdR on cell growth inhibition (P

Humans ; Infant ; Lead Poisoning, Nervous System, Childhood ; diagnosis ; therapy ; Male

Child ; Humans ; Lymphoma, T-Cell ; complications ; Panniculitis ; etiology ; Subcutaneous Tissue

Objective Inflammation and calcification are common phenomenons in human atherosclerotic plaque.The objective of this study was to determine the gene expression of some proteins and anti-inflammatory factors as well as bone formation mediators in the fibrous cap and shoulder region of non-calcified and calcified carotid endarterectomy(CEA) plaques.Methods Twenty-two CEA plaques were classified as Va(non-calcified,n=11) and Vb(calcified,n=11) in accordance with the AHA consensus in 1995.Mean percent in carotid stenosis and calcification area was 76.4% and 0% in Va and 74.2% and 33.6% in Vb respectively as determined by quantitative histomorphometry.Using laser capture microdissection,the fibrous cap and shoulder regions were excised from 22 frozen sections/plaques.After total RNA extraction and reverse transcription,gene expression of proteins(IL-1,IL-8,and MCP-1),anti-inflammatory factor(IL-10) and bone formation protein(BMP-6 and Osteocalcin) were detected by Real-time PCR.Results mRNA expression of MCP-1,IL-8 and IL-1 in Va plaques were 1.8 fold higher than that in Vb plaques,mRNA expression of anti-inflammatory factor IL-10 and osteogenic related genes BMP-6 and Osteocalcin were 3.8,4.2,and 6.9 fold higher in Vb plaques compared to Va plaques.Conclusions Our findings indicate that fibrous cap inflammation and susceptibility to disruption is more likely to occur in non-calcified plaques.

Objective To explore the effect of covered endoesophageal stent in treatment of terminal esophageal carcinoma complicated with malignant esophageal fistula in the elderly. Methods The covered endoesophageal stent was placed at the focus of lesion for each elderly patient in guidance by the iron wire and the stent-transporter under the endoscopy or X-ray. Results All of the stents were successfully implanted in 225 elderly patients without technical failure. The fistula was fully closed in all 19 patients. The symptoms of dysphagia and bucking were relieved obviously in 184cases (81.8%). The 176 cases (78.2%) of patients could have semi-fluid food in first week after stent implant, then have full meal. The inspiration pneumonia caused by fistula was brought under control. Conclusions For elderly patients with esophageal carcinoma, when they lose the operative opportunity or can not tolerate an operation, the treatment with covered endoesophageal stent is effective and safe.

ObjectiveUsing the technology of siRNA to inhibit the gene expression of no-receptor　tyrosine protein kinase Lck in T cells of asthmatic mice,and to study the therapeutic effect of Lck specific　siRNA in asthmatic mice.MethodsReceptor tyrosine protein kinase Lck specific siRNA fragments were　taken from chemosynthesis.In vivo-jetPEITM was used to transfect the siRNA into mice body through tail　vein injection.The mice were killed 48 hours later,and the levels of IL-4,IL-17 in bronchoalveolar lavage　fluid (BALF) were detected with respondent ELISA kits.The change of inflammatory histopathology in lung　was observed with H.E.staining.The expression of Lck in lung was detected with immunohistochemistry　(IHC),and the level of Lck in lung tissue homogenate was detected with Western Blot.Results Compared with asthmatic group[ (234.68 ± 11.15 ) pg/ml,( 96.76 ± 8.28 ) pg/ml],the levels of IL-4,IL-17　[ (234.68 ± 11.15)pg/ml,(96.76 ±8.28) pg/ml] in the BALF of siRNA interference group decreased,　and the inflammation in the lung relieved.IHC indicated that the expression of Lck in lung decreased and　the level of Lck in lung tissue homogenate decreased ( P ＜ 0.05 ).Conclusions Lck specific siRNA　could reduce the level of IL-4,IL-17 in the lung tissues of asthmatic mice,and relieve the inflammatory reaction in lung.

Objective To identify novel markers such as THRA or TCL1 in Sj(o)gren's syndrome (SS) patients to discriminate them from hcalthy volunteers.Methods Experimental group (n=40) and healthy volunteer group (n=40) were recruited based on strict inclusion and exclusion criteria.Peripheral blood samples (9 ml) were collected and peripheral blood mononuclear cells were separated by Ficoll gradient centrifugation.RNA was extracted using Trizol reagent,and the expression of THRA,TCL1 in PBMCs was detected by real-time PCR.And the data were processed with Wilcoxon test and t test (P＜0.05).Results TCL1 and THRA expression level are higher in SS patients (2.5±4.7) than healthy volunteers (Z=-2.045,P＜0.05).The TCLI expression level (4.4±3.8) is higher in high lymphadenopathy activation index patients (2 to 3grade) than that in low lymphadenopathy activation index patients (grade 0 to 1 ) (t=-2.319,P＜0.05 ).Conclusion TCL1 expression level is higher in SS patients,and TCL1 expression level is related to the severity of lymphadenopathy,which provide a new platform of the study for the pathogenesis,disease severity evaluation,even dia-gnosis and treatment of SS.

Objective Using the technology of siRNA to inhibit gene expression of T cells'nonreceptor tyrosine protein kinase Lck in asthmatic mice,and to study the effect of siRNA inhibited Lck to the function of T cells in asthmatic mice.Methods The 21 - 23 bp RNA fragments of mouse T cell Lck were made by chemosynthesis.INTERFERinTMsiRNA Transfection Reagent was used as transfection reagent to transfect the siRNA into the spleen T cells of asthmatic mice for 48 hours.Then T cells were mixed with bone marrow dendritic cells (DC) of asthmatic mice for another 48 hours.Cell culture suspension was collected and the level of IL-4,IL-13,IL-2,INF-γ were detected with respondent ELISA kits; Western Blot was used to identify if the expression of Lck was blocked.Results The expression of Lck in T cells almost could not be detected in siRNA interference group.The levels of IL-4 and IL-13 in siRNA interference group( 10.19 ± 1.66,12.34 ±0.79) were lower than no-siRNA interference(28.06 ±2.88,27.87 ± 1.61 )and control group ( 22.07 ± 2.5 1,20.47 ± 2.37 ),and the difference was statistical significant ( P ＜0.01 ).Conclusions Special siRNA could block the expression of special gene,and Lck specific siRNA could block the activation and differentiation of T cells and reduce the secretion of inflammatory cytokines in asthmatic mice.

BACKGROUND: Individual three-dimensional (3D) scaffolds can be constructed by 3D printing via Computer Aided Design based on the given anatomical measurements of related tissues. A rapid and accurate reconstruction of bone, cartilage, muscle and vessel also can be achieved by 3D printing; however, many problems still remain unsolved.OBJECTIVE: To summarize the principle and classification of 3D printing, the classification, characteristics and histocompatibility of scaffolds through reviewing the articles addressing 3D printing applied in bone tissue engineering,thereby providing theoretical foundation for the study on the construction of tissue-engineered bone.METHODS: PubMed and CNKI databases were retrieved for the literatures regarding the application of 3D printing technology in bone tissue engineering published from January 2001 to January 2017 using the keywords of three-dimensional printing, rapid prototyping manufacturing, bone tissue engineering in English and Chinese,respectively. Finally, 30 articles were reviewed and discussed in accordance with the inclusion and exclusion criteria.RESULTS AND CONCLUSION: The microstructures of normal tissues can be reconstructed and seed cells are printed on the 3D scaffolds synchronously by 3D printing technology. Moreover, the scaffold degradation and cell differentiation are synchronous, which contributes to tissue repair. Biological ceramics have been widely used in bone tissue engineering because of its good biocompatibility and mechanical properties. However, the urgent problems such as angiogenesis and cellular signal transduction still need to be addressed.

AIM: To evaluate the relationship between uPA and NF-?B p65 expression and its clinical significance in breast cancer. METHODS: The uPA mRNA was measured by real-time fluorescent quantitative PCR in 46 cases of breast cancer tissues and their adjacent counterparts. NF-?B p65 were measured using immunohistochemistry. RESULTS: The expression of uPA gene was elevated in 63% of cases, and there was a strong correlation between NF-?B p65 and uPA expression (r=0.451,P