OBJECTIVE:To establish the method for simultaneous determination of 9 components in Huichun yuzi granules. METHODS:LC-MS/MS method was adopted. The determination was performed on Shim-pack XR-ODS C18column with column temperature of 30 ℃. The sample size was 5 μL,and ion source as electrospray ion source;MRM mode was adopted. The acetonitrile-water was used as mobile phase for ferulic acid,rutin,paeonol,icariin and schisandrin(gradient elution);positive ion mode monitoring was conducted. The methanol-0.1% fomic acid water was used as mobile phase for naringin,verbascoside, amygdalin and protocatechuic acid(gradient elution);negative ion mode monitoring was conducted. RESULTS:The linear ranges of ferulic acid,rutin,paeonol,icariin,schisandrin,naringin,verbascoside,amygdalin and protocatechuic acid were 0.499 5-500, 25-1 000,0.245 9-250,5.185 1-1 000,0.981 9-1 000,0.248 1-125,2.510 4-250,10-2 500,4.999 7-1 000 ng/mL(r≥0.997 8), respectively. The limits of detection were 0.075,0.30,0.072,0.015,0.015,0.075,0.15,0.30,0.15 ng/mL,respectively;the limits of quantitation were 0.25,1.00,0.24,0.51,0.49,0.25,0.50,0.99,0.49 ng/mL,respectively. The recoveries rate were 91.39%-103.56%(RSD=1.03%-3.67%,n=6).RSDs of stability test ranged 1.4%-3.4%(n=6)within 48 h at room temperature. CONCLUSIONS:The method is rapid,sensitive and reproducible,and can be used for content determination of 9 components in Huichun yuzi granules.It can be used for quality control of Huichun yuzi granules.

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.

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.

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.

As food preferences and eating habits form early in life, the development of healthy eating habits in early childhood is a way to prevent diet-related diseases. The dietary pattern approach examines the effect of an overall diet on health outcomes, instead of individual foods or nutrients, thereby presenting a comprehensive evaluation of children's dietary intake. This article reviews the current literature to summarize the main methods for assessing dietary patterns and explore relationships between children's dietary patterns and obesity, puberty onset, cardiovascular diseases, and neurodevelopment. The purpose of this review is to provide evidence-based support for reducing the risk of diet-related diseases in children and recommendations for future research directions.
Child ; Child, Preschool ; Diet ; Eating ; Feeding Behavior ; Food Preferences ; Humans ; Obesity/prevention & control*