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Objective To analyze miR-92a expression and its clinical significance in the plasma of systemic lupus erythematosus (SLE) patients.Methods Plasma samples from 44 SLE patients,16 rheumatoid arthritis (RA) patients and 20 healthy controls were collected.The small RNAs in these plasma samples were isolated and reversely transcribed.Using cel-miR-39 as the external reference,the levels of miR-92a expression were detected by real-time polymerase chain reaction (PCR) method.MiR-92a and cel-miR-39 were analyzed by real-time fluorescence quantitative PCR and agarose gel electrophoresis.The sensitivity and specificity of miR-92a as SLE were analyzed by receiver-operating characteristic (ROC) curve.The correlation between the levels of miR-92a expression and the clinic pathological features of SLE and biological significance of miR-92a expression in SLE were further analyzed by Pearson or Chi-square test.Results Our data indicated that the plasma levels of miR-92a expression was 49.20 (5.33,95.17) in SLE patients,411.30 (320.84,504.69) in healthy controls,and 25.59(11.20,30.54) in RA patients.The difference was significant (x2=40.77,P＜0.01).The area under the ROC curve (AUC) was 0.958 for discriminating between SLE patients and normal subjects and 1.00 for discriminating between RA patients and healthy controls.The levels of miR92a expression cutoff values were set the as 198.59 for healthy control and 85.35 for RA patients,the diagnostic sensitivity and specificity were 93.2％,90％,and 100％,100％,res-pectively.The analysis of the correlation between miR-92a expression and the clinic pathological features of SLE had shown that the levels of plasma miR-92a expressions were much lower in SLE patients with down-regulated complement C3,and up-regulated urea nitrogen,creatinine,LDH,ATH (all P＜0 05).Conclusion Down-regulated miR-92a expression in plasma of SLE may be involved in the SLE disease occurrence or development and could be used as a novel potential diagnostic biomarker for SLE.
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Objective To investigate the polymorphisms of human cytomegalovirus ( HCMV ) UL146 gene in asymptomatic children. Methods Urine samples were collected from 47 asymptomatic chil-dren who were positive for HCMV DNA. PCR was performed to amplify the open reading frame ( ORF) of UL146 gene. Positive bands were sequenced and variations in UL146 gene were analyzed by using bioinfor-matics software. Results Seventeen samples were successfully amplified and sequenced. Variations spread all over the sequence of UL146 gene and the variability in nucleotide and amino acid sequences ranged from 0％ to 42. 5％ and 0％ to 67. 7％ respectively. Compared with the Towne strain, there was diversity in sig-nal sequence and C-terminal region. Phylogenetic analysis indicated that UL146 in the 17 asymptomatic chil-dren belonged to four genotypes, which were G1, G8, G9 and G11. Forms of post-translational modification varied greatly among the four genotypes, while the important functional region of ELRCXC chemokine was highly conservative. Secondary structure prediction showed that random-coli conformation was the predomi-nant structure of active proteins. Isoelectric point ( PI) and molecular weight ( MW) were dissimilar among the four genotypes. Conclusion HCMV UL146 gene in asymptomatic children was hypervariable in both nucleotide sequence and amino acid structure. However, the important functional region was highly con-served. The predominant genotypes of UL146 in these children were G1, G8, G9 and G11, and the geno-type distribution in them showed no significant difference with previous findings in children with symptomatic HCMV infection.
ABSTRACT
Objective To investigate the polymorphisms of human cytomegalovirus ( HCMV ) UL146 gene in asymptomatic children. Methods Urine samples were collected from 47 asymptomatic chil-dren who were positive for HCMV DNA. PCR was performed to amplify the open reading frame ( ORF) of UL146 gene. Positive bands were sequenced and variations in UL146 gene were analyzed by using bioinfor-matics software. Results Seventeen samples were successfully amplified and sequenced. Variations spread all over the sequence of UL146 gene and the variability in nucleotide and amino acid sequences ranged from 0％ to 42. 5％ and 0％ to 67. 7％ respectively. Compared with the Towne strain, there was diversity in sig-nal sequence and C-terminal region. Phylogenetic analysis indicated that UL146 in the 17 asymptomatic chil-dren belonged to four genotypes, which were G1, G8, G9 and G11. Forms of post-translational modification varied greatly among the four genotypes, while the important functional region of ELRCXC chemokine was highly conservative. Secondary structure prediction showed that random-coli conformation was the predomi-nant structure of active proteins. Isoelectric point ( PI) and molecular weight ( MW) were dissimilar among the four genotypes. Conclusion HCMV UL146 gene in asymptomatic children was hypervariable in both nucleotide sequence and amino acid structure. However, the important functional region was highly con-served. The predominant genotypes of UL146 in these children were G1, G8, G9 and G11, and the geno-type distribution in them showed no significant difference with previous findings in children with symptomatic HCMV infection.
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Objective To construct a recombinant adenovirus carrying UL138 gene, which was re-lated to the latent infection of human cytomegalovirus, and to investigate the effects of UL138 gene on the functions of THP-1 mononuclear cells. Methods The recombinant adenovirus expressing the UL138 gene was packaged. The titer of the recombinant adenovirus was determined by calculating 50% tissue culture in-fective dose ( TCID50 ) . THP-1 mononuclear cells were infected with the recombinant adenovirus at different multiplicity of infection (MOI) and the optimal MOI was determined (100 PFU/cell) by observing the ex-pression of green fluorescent protein ( GFP ) . Changes in the expression of proinflammatory cytokines by THP-1 mononuclear cells that was induced by overexpressed UL138 were analyzed by quantitative PCR. The expression of chemokines and their receptors were measured by quantitative PCR array. Results The re-combinant adenovirus carrying the UL138 gene was successfully constructed with a titer of 1×1011 PFU/ml. The rate of THP-1 mononuclear cells that was infected with the recombinant adenovirus was 60% at the MOI of 1 ∶ 100. Results of the RT-PCR analysis and Western blot assay further confirmed that the recombinant adenovirus could infect THP-1 mononuclear cells successfully and the expression of UL138 protein increased gradually over time. The overexpressed UL138 in THP-1 mononuclear cells significantly inhibited the expres-sion of IL-18, IL-1β, IL-6, IL-8 and TNF-α as indicated by the results of quantitative PCR. Results ob-tained from the quantitative PCR array analysis showed that most of the chemokines and their receptors were downregulated in the transfected THP-1 mononuclear cells except for the chemokines of CCL17, CCL21, CCL2 and XCL2 and the receptors of CCR2, CXCR1, CXCR2, CXCR4 and CX3CR1 which were upregulat-ed. Conclusion We successfully constructed the recombinant adenovirus carrying UL138 gene which could be used to infect THP-1 mononuclear cells. Overexpressed UL138 in THP-1 mononuclear cells significantly affected the functions of THP-1 mononuclear cells.
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Objective To investigate the effects of sodium ozagrel and berberine hydrochloride in the treatment of acute cerebral infarction and its effect on glucose and lipid metabolism.Methods According to the digital table,158 patients with acute cerebral infarction were randomly divided into the two groups:the control group was treated with the sodium ozagrel group and the observation group was treated combined with sodium ozagrel and berberine hydrochloride.After 2 weeks,Chinese stroke clinical neural function defect score scale and the National Institutes of Health Stroke Scale (NIHSS) was used to evaluate neurological function.The therapeutic effectiveness was assessed by ADL after treatment 1,3,6 months.The changes of glucose and lipid levels were detected by turbidimetric method.Results The total effective ratein the observation group was obviously higher than that of the control group (87.01％ vs 69.14％) (x2 =7.316,P ＜0.01).Of the above two ways of nerve function score indicateed that the observation group was significantly better than that of the control group (t =3.207,1.274,all P ＜ 0.01).ADL score of all patients was significantly better than that of before treatment with drug in the two groups after 1 month,3 months and 6 months (t =7.266 and t =8.850,t =1.429 and t =2.816,t =9.218 and t =6.739,P ＜0.01).Compared with the control group,ADL score of the observation group was significantly improved after 3 months and 6 months (t =1.163,0.932,all P ＜0.01).The significant difference was found between the control group and the observation group after 3 months and 6 months (t =7.861,5.005,all P ＞ 0.05).After the treatment,the fasting blood glucose level was reduced significantly in the observation group (t =0.859,P ＜ 0.01),total cholesteroland high-density lipoprotein,triglyceride,low density lipoprotein level were significantly decreased (t =0.257,0.114,0.378,all P ＜ 0.01),but there was no significant difference in HDL-C level (t =1.960,P ＞ 0.05).Conclusion The therapeutic effect of berberine hydrochloride combined with sodium ozagrel is better in the treatment of acute cerebral infarction,and the level of glucose and lipid metabolism was significantly decreased.