| Background and ObjectiveMetabolic syndrome (MetS) and type 2 diabetes mellitus (T2DM) are increasing metabolic disease in recent years in the whole whold especially in our developing country, including in China.Their incidence rate and mortality rate show a yearly increasing trend with changes in dietary structures and life styles of residents and prolongation of percapita expectancy in last decades. Insulin resistance is the most important pathological bases of obesity related metabolic disease.Metabolic syndrome (MetS) and type 2 diabetes mellitus (T2DM) are complex diseases that are affected by dietary intake and genetics. Hyperhomocysteinemia (Hhcy) has been proved is a risk factor for atherosclerosis and cardiovascular diseases. Two risk factors-lack of folic acid metabolism related diet and related genes mutations of Hhcy are becoming the high light cause research in MetS and T2DM. At the same time the development of nutrigenomics allows exploration of Hhcy clinical pharmacy intervention initiatives to reduce the metabolic syndrome, type 2 diabetes and cardiovascular diseases risk has become a widely important new topics.Dietary inadequacy can result in HHcy by presumably affecting the level of folate and vitamin B12. Methylenetetrahydrofolate reductase (MTHFR) is a key regulator for concentration of homocystine. MTHFR can irreversibly convert 5, 10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, which is the predominant form of folate found circulating in the blood and is the methyl donor during Hey remethylation. In addition, the activity of MTHFR is important for metabolism homeostasis. The C to T mutation of the MTHFR gene at nucleotide position 677 (MTHFR677>T) decreases its enzymatic activity, leading to accumulation of Hey. Such a mutation has emerged as a novel independent risk factor for metabolic disease, especially in Met,T2DM.The results of pathological and clinical studies suggest that inflammation plays a prominent role at every stage of atherogenesis. Increased levels of inflammatory proteins, such as hs-CRP, are predictive markers of an increased risk of cardiovascular diseases and are associated with atherosclerotic disease and insulin resistance.The relationship between the MTHFR C677T gene polymorphism and insulin resistance has not been previously reported. The association between MetS and the N5, N10-methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism, high-sensitivity C-reactive protein (hs-CRP) and the dietary components folate and vitamin B12 in Asians has not been well characterized. The purpose of our study is to explore distributions of MTHFR gene polymorphisms of folate metabolic enzymes in a Chinese population and associations between these polymorphisms and insulin resistance and susceptibility to Met and T2DM, and further to analyze the role of gene and environment interactions in the development of insulin resistance and risk factors of MetS.Materials and MethodsA total of 158 T2DM patients and 55 healthy subjects were enrolled in this study. The patients were enrolled in the department of Endocrinology of the Second Hospital of Lanzhou University between January 2007 and January 2008 were diagnosed with T2DM according to the criteria of the World Health Organization (WHO). The healthy participants were recruited during routine health screenings, had a normal glycemic index (3.3-6.1 mmol/L) and did not have metabolic diseases, such as hypertension, hyperlipidemia and obesity. All the subjects were of Han Chinese origin and did not have inflammatory diseases or cancers of any type.The MTHFR C677T gene polymorphism were analyzed by Polymerase Chain Reaction-Restriction Fragment Lenth Polymorphism (PCR-RFLP) in newly diagnosed T2DM patients with (118) and without MS (40) and in healthy subjects (55). Levels of insulin, folate and vitamin B12 by radioimmunoassay and of hs-CRP by immunoturbidimetry also were measured.All statistical analysis was processed by SPSS 13.0 for windows and Microsoft Excel 2003.Results1.MetS-associated T2DM accounts for 75%(118/158) of newly diagnosed 158 T2DM cases. Among these individuals,71.4%(70/98) are male and 80%(48/60) are female.2. A total of 158 cases of T2DM patients, aged 33-80 years.<50-year-old crowd MetS prevalence was 65.7%(23/35), while the population≥50 years old MetS prevalence was 76.4%(101/123). MetS prevalence increased with age, age≥60 years old, MetS prevalence rate of 74.1%(60/81), is<60 years people MetS prevalence rate was 35.1%(37/77) of the nearly 2-fold.3. The patients with MetS-associated T2DM had higher BMI, TCHO, TG, LDL, UA, FPG, SBP, DBP, HOMA-IR and hs-CRP than did their healthy counterparts (P<0.05). When compared to patients with non-MetS-associated T2DM, patients with MetS-associated T2DM had significantly higher levels of TCHO, TG, UA, blood pressure and HOMA-IR (P<0.05) and significantly lower levels of serum folate and vitamin B12 (P<0.05). In addition, patients with MetS-associated T2DM had higher BMI and hs-CRP levels in comparison to patients with non-MetS-associated T2DM; however, this difference was not statistically significant.4. The serum leve s of UA, TG, HOMA-IR and CRP were significantly higher in T2DM patients with a TT genotype than in those with a CC or CT genotype (P<0.05 for TG vs. CRP and P<0.001 for UA vs. HOMA-IR). In contrast, the levels of vitamin B12 in the serum of patients with a TT genotype were lower than those of patients with a CC or CT genotype (P<0.01).5.The frequencies of the MTHFR homozygous TT and heterozygous CT genotypes in T2DM patients diagnosed with MetS were 19.5% and 51.7%, respectively. These values were not only significantly higher than those in the healthy subjects (7.3% and 30.9%, respectively) but were significantly higher than those in non-MetS-associated T2DM patients (10% and 32.5%, respectively). The total T frequency of the MTHFR gene significantly increased from 22.7% and 26.3% in healthy subjects and non-MS-associated T2DM patients, respectively, to 45.3% in MetS-associated T2DM patients (x2=20.665, P<0.001)Conclusion1.The MTHFR C677T gene polymorphism may contribute to insulin resistance in Han Chinese individuals with MetS.2.MTHFR gene C677T polymorphism by increasing the levels of hs-CRP,TG and decreasing the levels of vitamin B12 in the serum of subjects with a TT genotype are related to MetS. As such, this polymorphism may play an important role in the development of MetS-associated T2DM.3.The total frequency of the T genotype significantly increased from 26.3% in T2DM patients without MetS to 45.3% in those with MetS.4. The T allele increases the susceptibility of a T2DM patient to MetS. Background and ObjectiveUrinary bladder cancer is the most common tumors,80% are urothelial cancers. Many patients with urothelial bladder carcinoma have distant metastasis when diagnosed, leading to surgical removal of poor prognosis. Therefore, it is necessary to the molecular mechanism of bladder cancer in-deep study to explore new methods of early diagnosis and treatment.MicroRNA(miRNA) is a new discovery of a length of 22 nucleotide non-coding single-stranded small molecule RNA, accounting for human gene 1%, it regulates at least 30% of the total mRNA. In recent years, the relationship of miRNA with tumor are becoming a hot spot. The role of miRNAs is similar to tumor suppressor gene and oncogene function, At present, research on miRNA and bladder cancer is very few, There is still no research on bladder cancer and microRNA expression profiles in paraffin embedded tissues are reported. We combine the foundation of previous work on bladder cancer research, to investigate miRNAs profile of bladder cancer in FFPE tissue for research materials.Materials and MethodsAll subjects including 103 patients with bladder cancer and other surgical resection of the bladder tissues from the Institute of Urology of Second Hospital of Lanzhou University, archive 1998 to 2000 and between 2004-2005 were fixed in formalin and embedded in paraffin.Microarray genechip technology and PCR are used to evaluation the effectiveness and feasibility of expression analysis miRNA from FFPE. Screening biological characteristics and differential expression of miRNA of bladder cancer. Results:1 microRNA difficult to degrade so can provide effective RNA requirements to extracted miRNA microarray;2 There are 15 upregulated genes were miR-141, miR-429, miR-200a, miR-205, miR-203, miR-200c, miR-190, miR-200b, miR-210, miR-10b, miR-425, miR-34a, miR-191, miR-31, miR-10a in bladder urothelial carcinoma compared to normal bladder epithelial tissues. Other three, miRNAs -miR-487a, miR-886-5p, miR-486-5p are downregulated genes.There are no relationship between miRNA changes and pathological stages.3 Microarray chip can effectively distinguish different stages of normal bladder tissue and tumor tissue.4. MiRNA-203 of FFPE bladder cancer was significantly increased 134-fold compared with the normal, and miRNA-141 was significantly increased 373-fold compared with the normal.5. MiRNA-200 family members (miRNA-200a, miRNA-200b, miRNA-200c, miRNA-141 and miRNA-429) of bladder cancer increased compared with normal tissues, miR-141, miR-429, miR-200a, miR-200b, miR-200c were raised over the normal 337,131,21,18,10 times.Conclusions1. Paraffin embedded tissue (FFPE) can be used as the effective detection of microRNA specimens;2. The storage time (10 years; 5-years)of FFPE tissue can effectively detect the miRNA;3 Eighteen miRNAs were significantly differentially expressed between bladder cancer versus normal tissues. Overexpression of fifteen, they are miR-141, miR-429, miR-200a,miR-205,miR-203, miR-200c, miR-190, miR-200b, miR-210, miR-10b, miR-425, miR-34a, miR-191, miR-31, miR-10a. The other three miRNA are downregulated, miR-487a, miR-886-5p, miR-486-5p.4. MiRNA-203 of FFPE bladder cancer was significantly increased 134-fold compared with the normal tissue, and miRNA-141 was significantly increased 373-fold compared with the normal. Therefore speculated that miRNA-203, miRNA-141 can be used to determine in the diagnosis of bladder cancer.5. MicroRNA have no related changes with pathological staging.6. MiRNA-200 family members (miRNA-200a, miRNA-200b, miRNA-200c, miRNA-141 and miRNA-429) may involved in bladder cancer.
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