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Objective To investigate the vitamin B 12 status of vegetarians in Shanghai.Methods A total of 282 adult vegetarians and 282 omnivores matching by gender and age were recruited in Shanghai.Their dietary intakes were collected.The serum concentrations of vitamin B12,folate and homocysteine were tested.The red blood cell,hematocrit value,mean corpuscular volume and mean erythrocyte width were also examined.Results The daily average intake of dietary vitamin B12 was (0.46± 1.01) μg/d in vegetarians and only (0.1±0.46) μg/d in vegans,which was lower than that of omnivores [(3.91±6.92) μg/d,F=50.57,P＜0.01].137 omnivores and 274 vegetarians had less dietary vitamin B12 level than recommended nutrient intake (RNI) and the difference was statistically significant (x2 =114.77,P＜ 0.01).54.26％ of vegetarians,68.92％ vegans,49.04％ ovo-lacto vegetarians and 15.60％ omnivores had hyperhomocysteinemia and the differences between vegetarians and omnivores were statistically significant (all P＜0.01).After adjusting the confounding factors the hematocrit value was higher in vegetarians,vegans and ovo-lacto vegetarians than in omnivores (27.42％± 18.32％,28.73％± 18.19％,26.95％± 18.38％ vs.8.96％± 16.59％,P＜0.01).Vegans had lower red blood cell counts and higher hematocrit value and mean corpuscular volume than omnivores.Conclusion Vitamin B12 deficiency combined with an elevated level of homocysteine and red blood cell volume growth are common but serious issue in vegetarians,especially in vegans.
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Objective@#To investigate the expression of miR-212 and miR-132 in the serum of patients with primary liver cancer and their targeted regulation of GP73.@*Methods@#The patients with liver cancer, chronic hepatitis B, or liver cirrhosis who were hospitalized in Taizhou People’s Hospital from January 2015 to December 2016 were enrolled, and healthy volunteers were also enrolled as controls. Quantitative real-time PCR was used to measure the serum levels of miR-212 and miR-132, and the association between the expression of serum miR-212 and miR-132 and the clinicopathological features of patients with liver cancer was analyzed. A Spearman’s rank correlation analysis was used to analyze the correlation between serum miR-212/miR-132 and GP73. Western blot was used to measure the protein expression of GP73, and MTT assay was used to measure the survival rate of cells. The Levene’s homogeneity of variance test was used for data analysis. The independent samples t-test was used for comparison of means between two samples, and ANOVA was used for comparison of means between multiple samples.@*Results@#A total of 90 patients with liver cancer, 60 with chronic hepatitis B, 68 with liver cirrhosis, and 100 healthy volunteers were enrolled. The relative expression levels of miR-212 and miR-132 in serum were 0.046 6 ± 0.024 7 and 0.005 9 ± 0.003 0 in the patients with liver cancer, 0.979 7 ± 0.259 5 and 1.001 8 ± 0.249 9 in the healthy volunteers, 0.588 2 ± 0.216 5 and 0.345 7 ± 0.233 8 in the patients with hepatitis, and 0.313 8 ± 0.153 3 and 0.080 1 ± 0.042 66 in the patients with liver cirrhosis. Compared with the normal controls, all patients had significant reductions in the expression of serum miR-212 (t = 10.26, 20.86, and 35.80, all P < 0.01) and miR-132 (t = 16.55, 36.09, and 39.85, all P < 0.01). In the patients with liver cancer, the relative expression of miR-212 and miR-132 was negatively correlated with alpha-fetoprotein (miR-212: t = -4.46, P < 0.01; miR-132: t = -4.83, P < 0.01), TNM stage (miR-212: t = 6.569, P < 0.01; miR-132: t = 7.31, P < 0.01), degree of tumor differentiation (miR-212: t = 5.268, P < 0.01; miR-132: t = 5.914, P < 0.01), and presence of portal vein tumor thrombus (miR-212: t = 5.16, P < 0.01; miR-132: t = 3.681, P < 0.01), while it was not correlated with tumor size (miR-212: t = 0.687, P > 0.05; miR-132: t = 0.887, P > 0.05). In addition, serum miR-212 and miR-132 were negatively correlated with GP73 in the patients with liver cancer (miR-212: rs = -0.709, P < 0.01; miR-132: rs = -0.877, P < 0.01). Overexpression of miR-212 or miR-132 in HepG2 cells significantly inhibited the activity and expression of 3’-UTR, and interference of miR-212 or miR-132 significantly increased the activity and expression of 3’-UTR in GP73. Overexpression of GP73 reversed the reduction in survival rate of hepatoma cells induced by the overexpression of miR-212 or miR-132.@*Conclusion@#Patients with liver cancer have a significant reduction in the expression of miR-212 and miR-132 in serum, which is closely associated with the development, progression, and metastasis of liver cancer, and miR-212 and miR-132 in hepatoma cells inhibit the growth of liver cancer by targeted regulation of GP73 expression.
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Objective To screen and analyze microRNA (miRNA) molecules associated with immune clearance in patients with chronic hepatitis B (CHB).Methods Twelve hospitalized CHB patients and 9 healthy individuals from Taizhou People's Hospital during June 2011 and August 2012 were enrolled.miRCURYTM chip was used to identify differential expression patterns of miRNA.The target genes of differentially expressed miRNA molecules were predicted by TargetScan,miRBase and miRanda databases,and their molecular pathways and functions were analyzed with bioinformatic methods.Results Compared with healthy individuals,52 differentially expressed miRNA molecules were identified in PBMCs of CHB patients,including 33 up-regulated and 19 down-regulated ones.With TargetScan,miRBase and miRanda databases,354 target genes were predicted in up-regulated miRNA molecules,and 1 935 target genes were predicted in down-regulated miRNA molecules.Gene ontology (GO) and Pathway analysis showed that several molecular pathways might be affected by up or down-regulated miRNA molecules.miRNA-mRNA network analysis showed that some target genes might be regulated by miRNA molecules like hsa-miR-520d-5p,hsa-miR-106a-Sp,hsa-miR-30a-5p and hsa-miR-29b-3p,and constituted a complex molecular network.Conclusion There are several miRNA molecules with abnormal expressions in CHB patients,which may be involved in immune clearance through the regulation of target genes and molecular pathways.
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Objective To explore the expression profiles and their clinical significance of microRNA (miRNA) molecules in different stages of chronic hepatitis B virus (HBV) infection.Methods The miRNA molecule expressions of 12 patients with chronic hepatitis B,12 chronic HBV carriers,12 inactive hepatitis B surface antigen (HBsAg) carriers,and 9 healthy controls were screened using miRNA microarray.The miRNA profiles were validated by the real time fluorescence quantitative polymerase chain reaction (PCR).The t-test was used for comparison between twogroups,whereas one-way ANOVA and SNK-q tests were used for multiple comparisons.Mann-Whitney and Kruskal Wallis H tests were used for comparison of two or more groups of data with skewed distribution.The receiver operation characteristic (ROC) curve was constructed to evaluate the diagnostic significance of miRNA molecules.Results Compared with the healthy controls,significant differences in the expression profiles of miRNA molecules were found in peripheral blood mononuclear cells of chronic HBV carriers (2 molecules up-regulated,and 18 down regulated) and chronic hepatitis B patients (33 molecules up-regulated and 19 down-regulated).No significant difference was found between inactive HBsAg carriers (2 molecules up regulated,and 3 down-regulated) and controls.The results of six miRNA molecules detected by real-time fluorescence quantitative PCR were basically consistent with the results detected by microarray.The area under ROC curve of the six miRNA molecules of hsa miR-4711-3p,hsa-miR-3191 5p,hsa-miR-5704-5p,hsa-miR 548ah-5p,hsa-miR-146a-5p and hsa-miR-29b-3p in distinguishing immune tolerance and clearance of chronic HBV infection were 0.994,0.984,0.967,1.000,1.000 and 0.996,respectively.Conclusions The different expression profiles of miRNA molecules could be used to distinguish the different stages of chronic HBV infection,and are closely related with the immune tolerance and activation in chronic HBV infection.