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 To detect the type of gene mutation of thalassemia in Kunming city.Methods Sixty-three cases highly suspec-tive of thalassemia were determined with the methods of ploymerase chain reaction(PCR) and reverse dot blot(RDB) for the type of gene mutation.Results According to gene analysis,35 cases were final diagnosed from 63 cases suspective of thalassemia.Among the total,4 cases were gene deficiency ?-thalassemia,and 30 cases were gene deficiency ?-thalassemia,and there was 1 case both ?-thalassemia and ?-thalassemia.There were 9 types of gene mutation with 15 gene combinations in 35 samples.The main type of ?-thalassemia was--SEA/??,there were 6 types with 11 gene combinations from the types of genes of ?-thalassemia,the highest incidence of gene mutation was 17 site,including 17 site homozygote,heterozygote and double heterozygote.Conclusions The thalassemia invasion of Yunnan has its characters,and it is valuable to launch further research.In the same patient,there are ?-thalassemia and ?-thalassemia,it signifies that those 2 types should be diagnosed in the same time,to prevent missed diagnosis.

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.

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 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.

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 To study the effects of semiconductor laser irradiation and high voltage static electric fields on small vessel damage in diabetic rats.Methods Fifty healthy male Wistar rats were randomly divided into two groups:8 rats in a normal group and 42 in a diabetic model group.The diabetic models were created by intrape- ritoneal injection of streptozocin.The diabetic model rats were randomly divided into four subgroups:a diabetes group,a semiconductor laser treatment group,a high voltage static electric field treatment group and a comprehensive treatment group receiving combined semiconductor laser and high voltage static electric field exposure.The rats in each treatment group were subjected to the corresponding intervention.After 20 days of treatment,the venous blood, kidney tissue and myocardium tissue of the rats were collected,and the concentrations of blood glucose,insulin,en- dothelin and tissue were detected.Results Compared with the normal group,a significant increase in blood glu- cose and endothelin was observed in the diabetic model group,along with significantly decreased blood insulin and significant small vessel endothelium proliferation in the kidney tissue.Compared with the diabetes group,endothelin levels were significantly lower in all 3 treatment groups,and blood insulin was also higher in the comprehensive treat- ment group.Conclusion There were severe abnormalities in blood glucose,insulin and endothelin as well as mild impairment of small vessel endothelium proliferation in the diabetic rats.Semiconductor laser and high voltage static electric field exposure have a role in treating and preventing these conditions in diabetic rats.