[Determination of six bisphenols in human urine by ultra-high performance liquid chromatography-tandem mass spectrometry].

OBJECTIVE: To develop and validate a solid phase extraction-ultra-high performance liquid chromatography-tandem mass spectrometry method for the determination of six bisphenols(bisphenol S, bisphenol F, bisphenol A, 2, 2'-methylenediphenol, bisphenol AF, bisphenol AP) in urine. METHODS: After enzymolysis of urine sample, the target substances were quickly purified and extracted by WAX solid phase extraction column. On ACQUITY BEH C_(18) column(2.1 mm×100 mm, 1.7 µm), the mobile phase of water and methanol was used to separate. Finally, multi-reaction detection was carried out under electrospray negative ion scanning, and quantification was carried out by internal standard method. RESULTS: The correlation coefficients(r) of the target compounds were all more than 0.998 in the range of 0.1-50.0 ng/mL, the linearity was good, and the detection limits were all lower than 0.1 ng/mL. The recoveries of the three standard concentrations(0.5, 5.0 and 50.0 ng/mL) were all between 80% and 120%, and the relative standard deviation was less than 20%(n=5). The standard reference material was detected and the concentration was within the reference range. CONCLUSION: This method can be used to detect six bisphenols in urine quickly and accurately, is suitable for the trace analysis of bisphenol compounds in human urine.

[Association of polymorphisms in SEC16B, DNAJC27, FTO and MC4R genes with overweight and obesity in Han preschool children].

OBJECTIVE: To investigate the association of polymorphisms in SEC16B rs633715, DNAJC27 rs713586, FTO rs11642015 and MC4R rs6567160 with overweight and obesity in Han Chinese preschool children. METHODS: A total of 749 Han Chinese preschool children from Henan and Guizhou Province of Long-term Health Effects Assessment Project of Infants and Toddlers Nutritional Pack were selected for the study and divided into an overweight and obese group and a normal control group in 2022. rs633715, rs713586, rs11642015 and rs6567160 were genotyped using Kompetitive allele-specific PCR(KASP) technology. The distribution of genotypic polymorphisms was compared using the χ~2 test. The association between the four loci and overweight and obesity in preschool children was analyzed using a multifactorial logistic regression model. RESULTS: The statistical analysis revealed a significant disparity(P<0.05) in the distribution of genotypic polymorphisms of rs633715 and rs6567160 among preschoolers in Henan and Guizhou Province. CC heterozygous mutant and recessive models at rs633715 locus were associated with susceptibility to overweight and obesity in preschool children [OR and 95% CI 2.915(1.163-7.305), and 2.997(1.226-7.323), respectively, both P<0.05]. TC heterozygous mutant and dominant models at rs713586 locus were also associated susceptibility to overweight and obesity in preschool children(OR and 95% CI were 2.362(1.054-5.289)and 2.362(1.054-5.289), respectively, both P<0.05). rs11642015 and rs6567160 loci were not associated with susceptibility to overweight and obesity in preschool children(P>0.05). The result of the analysis of the cumulative effect of rs633715 and rs713586 showed that the number of genotypes carrying the risk genotype was positively associated with the risk of overweight and obesity in preschool children(P_(trend)<0.01). CONCLUSION: Among Han Chinese preschool children, SEC16B rs633715 and DNAJC27 rs713586 were associated with susceptibility to overweight and obesity in preschool children. Moreover, rs633715 and rs713586 had a cumulative effect on susceptibility to overweight and obesity in preschool children, the number of risk genotypes carried was positively associated with childhood overweight and obesity risk.

[Prediction equation of physical activity level based on step counts for Chinese adults: development and validation].

OBJECTIVE: To develop a prediction equation for estimation physical activity level(PAL) based on the average daily steps in Chinese adults and to verify its prediction power. METHODS: A total of 800 adults with no restriction on physical activity were recruited. Excluding those who wore Actigragh WGT3X-BT accelerometer continuously for less than 3 days, 743 subjects(324 males, 419 females, mean age 31.4 years) were finally included. Daily steps, time of low intensity physical activity(LPA), moderate physical activity(MPA) and vigorous physical activity(VPA) and physical activity energy expenditure(PAEE) were measured by the accelerometer. The method of external validation was adopted. The measured PAL calculated by basal metabolic rate(BMR) measured by indirect thermography(IC) and total energy expenditure(TEE) measured by doubly labeled water method was used as the gold standard. The consistency of the prediction formula was verified by Bland-Altman analysis and the accuracy was verified by the accurate. RESULTS: The prediction equation was developed: PAL=1.17 + 0.000028 Steps. In the external verification, the mean difference was 0.05, the 95% consistency limit was(-0.26, 0.36), and the accuracy was 73%, indicating a good consistency. CONCLUSION: The prediction formula of PAL for Chinese adults based on the steps has good performance and can be applied to the management of physical activity and the prevention of obesity in Chinese adults.

[Serum metabolomics of iron deficiency anemia in infants based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry].

OBJECTIVE: To explore the differential metabolites in the serum of infants with iron deficiency anemia(IDA) and non iron deficiency anemia, and to explore the potential biomarkers. METHODS: Non-targeted metabolomics of 30 infants with iron deficiency anemia aged 6-11 months and 30 infants with non iron deficiency anemia aged 6-11 months were analyzed by ultra-high performance liquid chromatography with time of flight mass spectrometry using Acquity UPLC BEH C_(18) column(100 mm×2.1 mm, 1.8 µm). The differences of metabolites between the two groups were analyzed by principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). Differential metabolites were screened according to OPLS-DA variable importance projection(VIP) >1. The related metabolic pathways involved in the markers were analyzed based on the KEGG database. RESULTS: Differences in serum metabolic profiles between iron deficiency anemia group and non iron deficiency anemia group were observed. The 44 potential biomarkers were mainly lipids. Combined with pathway analysis, the metabolic pathways related to different metabolites included glycerophosphingolipid metabolism and sphingolipid metabolism. CONCLUSION: There are differences in lipid metabolites between infants with non iron deficiency anemia and iron deficiency anemia, suggesting that the occurrence and progress of iron deficiency anemia are related to lipid metabolism.

[Optimization of three-dimensional culture conditions of L02 cells with response surface methodology based on VitroGel system].

OBJECTIVE: This study optimizes three-dimensional(3D)culture conditions of L02 cells using response surface methodology(RSM) based on the VitroGel system to construct the hepatocytes model in vitro. METHODS: L02 cells were 3D cultured by the VitroGel system. The appropriate level of three key factors(concentration of inoculated cells, culture time and dilution degree of the hydrogel) was determined by single-factor experiment, and the optimal conditions of 3D culture of L02 cells based on the VitroGel system were determined by RSM. During the detection process, the optical density(OD) value of cell viability was used as the detection index, and the cell viability was detected using the cell counting kit-8(CCK-8) assay. The proliferative performance and viability of L02 cells was measured by fluorescent staining assay. RESULTS: The selected optimal culture conditions by RSM were as follows: concentration of inoculated cells was 1.1 × 10~5/mL, culture time was 9.5 days, and dilution degree of hydrogel was 1∶3.7. The result shows that under optimal conditions, the predicted OD value of cell viability was 2.17 and measured 2.13 with a relative error of 1.84%, indicating that the condition was suitable and reliable. The fluorescent staining and dead and live cells detection results showed the 3D hepatocytes model was successfully constructed. CONCLUSION: The optimal conditions for 3D culture of L02 cell based on the VitroGel system were determined by RSM, and a hepatocytes model with high cellular activity was successfully constructed.

[Gut microbiota in Tibetan, Mongolian and Zhuang children aged 10-12 years old in 2016].

OBJECTIVE: To study the composition of gut microbiota of Tibetan, Mongolian and Zhuang children. METHODS: A total of 100 school-age children aged 10-12 were randomly selected from three ethnic groups in 2016, the Tibetan in Bayi County, Tibet, the Mongolian in Zha Qi, Inner Mongolia, and the Zhuang in Shanglin County, Nanning, Guangxi. Fecal samples were collected for high-throughput sequencing and bioinformatics analysis to compare the structural differences of gut microbiota of children from the three ethnic groups in the above areas. RESULTS: The result of α diversity analysis showed that the diversity and richness of gut microbiota of children from three ethnic groups in the surveyed area were different in pairwise comparison. The dominant phyla and genera of Tibetan children's gut bacteria are Firmicutes(54.01%), Bacteroidetes(31.13%), Prevotella(15.72%) and Faecalibacterium(15.46%). The dominant phyla and genera of Mongolian children's gut bacteria are Firmicutes(54.89%), Bacteroidetes(17.85%), Proteobacteria(15.99%), Faecalibacterium(20.84%) and Escherichia(17.52%). While in Zhuang children, the dominant phyla and genera are Firmicutes(50.86%), Bacteroidetes(34.74%), Prevotella(18.40%), Bacteroides(17.78%) and Faecalibacterium(14.68%). CONCLUSION: The composition and structure of gut microbiota of Tibetan and Zhuang children in the three regions are similar, while the structure of gut microbiota of Mongolian children is obviously different from those of the first two regions.

MicroRNAs miR-18a and miR-452 regulate the replication of enterovirus 71 by targeting the gene encoding VP3.

MicroRNAs (miRNAs) are crucial in the process of host-pathogen interaction. In this study, we established a screening system for miRNAs of target genes to detect the effect of miRNAs on Enterovirus 71 (EV71) replication in rhabdomyosarcoma (RD) cells. A 3'-untranslated region (UTR) dual-luciferase assay was performed to confirm putative miRNA targets in EV71 genome. Firstly, 13 fragments of EV71 genome were inserted into the vector pMIR, and luciferase activities were analyzed to identify the putative miRNAs of target genes. The expression of the reporter protein was significantly downregulated in cells transfected with the vector containing gene VP3. Then we screened for miRNAs that might target to VP3 through online analysis software. In addition, Western blot, real-time PCR, virus titration, and morphological changes were considered to examine the effects of miRNAs on virus replication. The results suggested that miR-18a and miR-452 repress the reproduction of EV71 virus by binding to VP3. Moreover, EV71 infection also affected the expression of endogenous miR-18a and miR-452. In addition, no significant cytotoxic effects were observed. The results from this study suggest that the intracellular miRNAs may play vital roles in the host-virus interaction.

[Mechanism of microRNA-29a regulating steatosis in the human hepatocyte via targeting silent mating type information regulation 2 homolog-1].

OBJECTIVE: To investigate the expression of microRNA-29 a(miR-29 a) in the human steatotic hepatocyte model and the mechanism of targeting silent mating type information regulation 2 homolog-1(Sirt1)to regulate fat deposition of steatotic hepatocyte. METHODS: The nonalcoholic fatty liver cell model was induced by a mixture of oleic acid and palmitic acid. After successful validation model, the expression of miR-29 a and Sirt1 was measured by PCR. The target genes of miR-29 a was predicted in biological system. MiR-29 a mimic and miR-29 a inhibitor were transfected into hepatocytes, and then established the human steatotic hepatocyte model, the result of oil red O staining and triglyceride(TG)lipid content were observed, the expression of Sirt1 mRNA and protein were detected by qRT-PCR and Western blotting, respectively. RESULTS: The steatosis hepatocyte model was successfully established. Compared with control group, the relative expression of miR-29 a and triglyceride increased significantly(P<0. 01), while the relative expression of Sirt1 decreased significantly(P<0. 01) in the model group. Sirt1 was a target gene of miR-29 a. After transfection, the lipid droplet and the deposition of fat increased obviously in miR-29 a mimic group than those in the control group. TG content in miR-29 a mimic group increased significantly(P<0. 05), the expression of miR-29 a increased significantly(P<0. 01), while the expression of Sirt1 mRNA decreased significantly(P<0. 05), and the expression of Sirt1 protein showed a downtrend. On the contrary, after the inhibition of miR-29 a expression, the lipid droplets in miR-29 a inhibitor group were relatively reduced, the fat deposition was alleviated. The TG content was significantly decreased(P<0. 05), the expression of miR-29 a in the cells was effectively inhibited(P<0. 01), while the expression of Sirt1 mRNA was significantly increased(P<0. 05), and Sirt1 protein was on an upward compared with the control group. CONCLUSION: The expression of miR-29 a is significantly increased in the nonalcoholic fatty liver cell model. Upregulation of miR-29 a negatively regulates the expression of Sirt1, thus promoting fat deposition of steatotic hepatocyte.

[Energy and macronutrient intakes of employees at different levels of work intensity in China from 2010 to 2012].

OBJECTIVE: To analyze the intake of energy and macronutrients of employees at different levels of work intensity in China. METHODS: Based on the data of National Nutrition and Health Survey in 2010-2012, this paper analyzed the energy and macro nutrients intakes of employees at three levels of working strength. RESULTS: The energy intake of employees in China was 1952. 7 kcal/d, the intakes of protein, fat and carbohydrate were 62. 7 g/d, 76. 6 g/d and 254. 0 g/d, respectively, the energy supply ratios were 13. 1%, 34. 9% and 52. 4%, respectively. From food sources of energy, the percentage of total energy provided by cereals and animal foods was 47. 7% and 18. 2%, respectively. The percentage of protein from cereal, legumes and Animal food was 39. 8%, 6. 7% and 37. 5%, respectively. The percentage of fat from animal food was 34. 8%. From the age and work intensity groups, the energy intake was lower in the low age group and the light work intensity of employees. There was the lowest intake of protein and fats in high age groups with heavy work intensity. Energy levels form protein and fat were highest in the light intensity group. Energy supply from cereal food was the highest in the group with heavy work intensity and energy supply from animal food was the highest in the group with low age group and light work intensity. The intake of high quality protein and animal food fat decreased with the increase of age and work intensity. CONCLUSION: The energy intake of employees in China is lower than the recommended energy intake for Chinese residents, and fat provides a higher proportion of energy. Employees with high age and heavy work intensity have insufficient protein intake, and the unreasonable dietary structure was particularly prominent with them, and their nutritional status needs to be paid more attention.

[Validity of the international physical activity questionnaire and bouchard diary in Chinese adults].

OBJECTIVE: Based on the accelerometer, the validity of the international physical activity questionnaire long version(IPAQ-L) and Bouchard diary were evaluated to measure the daily physical activity of Chinese adults aged 18-59 years old. METHODS: A total of 200 Chinese adults were recruited in Yinzhou District of Ningbo City in 2019, including 78 males and 122 females, the three age groups 18-29, 30-49, 50 and above accounted for 19. 5%, 61. 5% and 18. 0%, respectively. The volunteers wore Actigraph WGT3 x-BT three-dimensional accelerometer for 3 days, and completed the 3 consecutive days& apos; Bouchard diary and the IPAQ-L. The physical activity energy expenditure(PAEE), sedentary inactivity time, light physical activity time(LPA) and moderate-to-vigorous physical activity time(MVPA) measured by the three tools were estimated respectively. Spearman analysis was used to analyze the correlation, the Friedman test and the Bland-Altman plot were used to test the homogeneity of the three method. RESULTS: 196 volunteers were included in this study. The correlation coefficients of PAEE between IPAQ and diary and accelerometer were 0. 32 and 0. 58, respectively, the correlation coefficients of sedentary behavior were 0. 17, 0. 25, and LPA time was 0. 33, and the estimated IPAQ for MVPA time better than diary. The difference test result showed that, except for IPAQ in the measurement of MVPA, there was no statistical difference from the accelerometer measurement result(P=0. 684), and the difference between other estimated values and the accelerometer measurement result was statistically significant(P& lt; 0. 01), Bland-Altman plot result showed that the diary and IPAQ had good consistency in the measurement of PAEE and IPAQ in the estimation of MVPA time, but there was an overestimation in PAEE. In addition, IPAQ had poor validity in measuring sedentary behavior and LPA. CONCLUSION: IPAQ and Bouchard diary have better validity in evaluating PAEE, and the diary is better than IPAQ. IPAQ has better validity in evaluating MVPA time, but has poor validity in estimating sedentary behavior and LPA time.

[Energy and macronutrient intakes of farmers at different levels of physical activity in China from 2010 to 2012].

OBJECTIVE: To analyze the intake and structure of energy and macronutrients of farmers under three kinds of job types: working, farming and housework in China, and analyzes the intake status under different work intensity. METHODS: Based on the data of China Nutrition and Health Survey in 2010-2012, this paper analyzed the dietary intake of farmers over 18 years old at three job types and different work intensities. RESULTS: The energy intake of farmers in China was 2149. 5 kcal/d, and that of men and women were 2345. 2 kcal/d and 1985. 4 kcal/d, respectively. Farming farmers took in the highest energy and the domestic farmers took in the lowest energy. The average protein intake of farmers was 60. 8 g, 66. 0 g for men and 56. 4 g for women. The protein intake of working, farming and housework farmers, showed a downward trend. The fat intake of farmers was 66. 6 g, including 71. 9 g for men and 62. 1 g for women. With the three main forms of working, farming and housework, the fat intake showed a downward trend. The carbohydrate intake of farmers was 327. 5 g, including 354. 6 g for men and 304. 7 g for women, with farming farmers taking in the highest carbohydrate. Among the working farmers, under the light, medium and heavy work intensity, the intake of carbohydrate increased in turn, but protein and fat had no obvious characteristics; among the farming workers, under the light, medium and heavy work intensity, carbohydrate, protein and fat showed an increasing trend. There were also differences in the dietary structure characteristics among the three job types of working, farming and housework. The proportion of energy from protein and fat was higher in working farmers, the proportion of high-quality protein was close to 35%, the proportion of energy from carbohydrate was the highest in farming farmers, the proportion of high-quality protein was only 27%, the proportion of high-quality protein in household farmers was 30%, and the ratio of fat to energy in both farming and household farmers was less than 30%. Among working farmers, under the light, medium and heavy work intensity, the proportion of energy from carbohydrate increased, while the proportion of protein from animal food decreased; among farming farmers, under the light, medium and heavy work intensity, the proportion of protein from animal food increased slightly. CONCLUSION: There are some differences in the energy and macronutrients intake among the working farmers, farming farmers and household farmers in China. The farming farmers have the most energy intake, and with the increase of work intensity, the proportion of carbohydrate intake increases, and the protein intake is insufficient, especially the heavy work intensity. The nutrition needs of the population should get more attention.

[Construction and identification of stable hFcεRIαßγ/RBL-2H3 cells for food allergy assessment].

OBJECTIVE: To establish an rat basophil leukemia(RBL)-2H3 cell line stably expressing human high affinity receptor containing alpha, beta and gamma chain(hFcεRIαßγ), in order to provide experimental materials for evaluating allergenicity of food. METHODS: The lentivirus was transfected into RBL-2H3 cells, and the mRNA expression of hFcεRIαßγ in cells was detected by real-time PCR and the protein expression of hFcεRIα was detected by flow cytometry. RESULTS: Sequencing result showed that recombinant lentiviral vector GV367-hFcεRIαßγ was successfully constructed. According to the result of experiments, lentivirus could effectively infect RBL-2H3 cells. The mRNA of hFcεRIαßγ and protein levels of hFcεRIα in RBL-2H3 cells were successfully overexpressed. CONCLUSION: The hFcεRIαßγ/RBL-2H3 cells were preliminarily constructed, which could be binded with human IgE and further used in the evaluation system of food allergy, compared to RBL-2H3 cells.

[Effects of nutrition package on intestinal flora of infants by high-throughput sequencing].

OBJECTIVE: To investigate the effect of nutrition package on intestinal flora of infants aged 6-24 months. METHODS: A total of 118 infants in Songxian County and Ruyang County of Henan Province, 60 infants in Songxian County with nutrition package coverage, and 58 infants in Ruyang County of the control county were selected for the long-term effects of early childhood nutrition package intervention project in 2018. All infant faeces were collected, and the composition and abundance of intestinal flora were analyzed and compared by high-throughput sequencing technology. RESULTS: The Alpha diversity of intestinal flora in the nutrition package intervention group was significantly higher than that in the control group. Firmicutes were on the rise in the nutrition package intervention group of infants aged 6-24 months. The abundance of Prevotella in the intervention group was significantly lower than that in the control group. The abundance of Faecalibacterium and Bacteroides in the nutrition package treatment group for infants aged 12 to 17 months was significantly higher than that of the control group, while the abundance of Bifidobacterium and Prevotella was significantly lower than that of the control group. Lactobacillus was significantly less abundant in the 18-24 month old infant nutrition package intervention group than in the control group. CONCLUSION: Nutrition package can improve the diversity of intestinal flora, also play an important role in promoting the balance of intestinal flora of infants.

[Immune function effect of F3 rats fed with genetically modified maize harboring Cry1Ab and epsps genes].

OBJECTIVE: To evaluate the effects of genetically modified maize with Cry1Ab and epsps genes on immune function in F3 rats. METHODS: A total of 180 weaning SD rats for F0 generation were randomly divided into three groups, which were treated with AIN-93 G feed control diet, parental maize diet and genetically modified maize diet respectively. After three generations of breeding, antibody producing cells determination, concanavalin A(ConA)-induced lymphocyte transformation test, natural killer(NK)cells activities assay, whole blood lymphocyte subtype detection, delayed type hypersensitivity test and immunity organ index were performed. RESULTS: There were no significant differences between parental maize diet and genetically modified maize diet in terms of the number of antibody-producing cells, ConA-induced spleen lymphocyte proliferation, NK cell activity, whole blood lymphocyte subsets, delayed type hypersensitivity and thymus index(P>0. 05). CONCLUSION: Under the conditions of this experiment, no significant effects were found on immune function of F3 SD rats through the three generation development study of genetically modified maize with CrylAb and epsps genes.

[Assessment of allergenicity of oryza sativa recombinant human serum albumin in BALB/c mice].

OBJECTIVE: To investigate the potential allergenicity of oryza sativa recombinant human serum albumin(OsrHSA)in BALB/c mice. METHODS: Eighty BALB/c mice were randomly divided into four groups i. e ovalbumin(OVA) positive control group, potato acid phosphatase(PAP) negative control group, Oryza sativa recombinant HAS(OsrHSA) group and solvent control group(phosphate buffer saline, PBS), respectively. Mice were administered by intraperitoneal injections of tested proteins and histamine levels in plasma and sIgE, sIgG, sIgG1, sIgG2 a, and tIgE antibody levels in serum were measured. RESULTS: Compared with the other groups, serum tIgE, sIgE, sIgG, sIgG1 and plasma histamine levels in the OVA group were significantly increased, while serum sIgG2 a levels were decreased(P<0. 05). There was no significant difference in serum sIgE level and histamine level between the OsrHSA group and the control group(P>0. 05). Serum sIgG, sIgG1 and sIgG2 a levels were lower than those in the PAP group(P<0. 05). There was no significant difference in serum tIgE content between PAP group and OsrHSA group(P>0. 05). CONCLUSION: The potential allergenicity of OsrHSA through traperitioneal injection in BALB/c mice was very low.

[Determination of flavonoids in honey by solid phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry method].

OBJECTIVE: To establish a method for simultaneous determination of 20 flavonoid compounds from honey samples using solid phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS), which was applied to study the difference of flavonoid compounds in honey samples from different botanical origins. METHODS: The honey samples were extracted with ultra-water solution by ultrasound, and purified with an Oasis MAX column, and then separated on a Waters ACQUITY UPLC HSS T3(2. 1 mm×100 mm, 1. 8 µm) and eluted with the mobile phase consisted of 0. 1% formic acid in water and acetonitrile in a gradient program, and analyzed by mass spectrometry equipped with an electrospray ionization source, operating in the positive or negative ionization modes, and performed in the multiple reaction monitoring using external standards. RESULTS: Twenty flavonoid compounds showed good linearity with coefficients higher than 0. 9969; The recoveries of spiked honey samples in the range of 76. 2%-108. 6% at the concentrations of 100, 250 and 500 µg/L; The relative standard deviations were 0. 50%-4. 34% and 2. 51%-6. 71%(n=6)for inter-day and intra-day precisions, respectively. It was found that the variations of flavonoid compounds in 16 types of honey samples from different botanical origins differed largely. Among them, quercetrin content in fennel honey was the highest(165 ng/g). High contents of luteolin in red eucalyptus and manuka honey were 186 ng/g and 304 ng/g, respectively. High contents of kaempferol in buckwheat, fennel and manuka honeys were from 113 to 257 ng/g. High contents of chrysin in snow lotus honey, manuka, sunflower honey and lavender Honey were from 136 to 213 ng/g. The content of pinocembrin(473 ng/g) and pinobanksin(684 ng/g) in manuka honey were much higher than that in other honeys. CONCLUSION: The proposed method is simple for pretreatment, also sensitive, accurate and producible for quantitative of 20 flavonoid compounds in different honey samples, which provide more scientific data and application basis for quality analysis.

[Steatosis and inflammation of L02 hepatocytes induced by sodium oleate].

OBJECTIVE: To observe the steatosis and inflammatory effects of L02 hepatocytes induced by different concentrations of sodium oleate. METHODS: L02 cells were incubated with sodium oleate in different concentration for 24 h, and the cell viability was detected. L02 cells were respectively cultured with 75, 150, 300 µmol/L sodium oleate for 24 h. The lipid accumulation of the cells was observed by oil red staining. The content of triglyceride in the cells was detected, the IL-6 content in the cell supernatant was detected. The expression of SIRT1 and nuclear factor-κB(NF-κB) was detected by Western Blot. The expression of Toll-like receptor 2 and Toll-like receptor 4(TLR2 and TLR4) on the cell surface was detected by flow cytometry. RESULTS: With the increase of sodium oleate concentration, the cell viability decreased, the cell growth inhibition rate increased. The content of triglyceride in L02 cells treated with 75, 150 and 300 µmol/L sodium oleate was significantly higher than that in the control group(P<0. 01, P<0. 001, P<0. 001), the IL-6 in the supernate was significantly higher than that in the control group(P<0. 05, P<0. 01, P<0. 001). The result of oil red staining showed that the lipids in the cells were obviously accumulated after sodium oleate treatment. The expression of TLR2 in L02 cells treated with 150 and 300 µmol/L sodium oleate was significantly higher than that in the control group(P<0. 05, P<0. 001). There was no significant increase in TLR4 expression after sodium oleate intervention in L02 cells. The expression of SIRT1 protein in the sodium oleate group was lower than that in the control group, and the expression of NF-κB p65 was higher than that in the control group through Western Blot result. CONCLUSION: L02 hepatocyte steatosis caused by sodium oleate may be associated with TLR2/NF-κB mediated inflammatory pathway.

Basal energy expenditure, resting energy expenditure and one metabolic equivalent (1 MET) values for young Chinese adults with different body weights.

BACKGROUND AND OBJECTIVES: Accurately assessing energy expenditure (EE) of people with different body weight is essential to facilitate weight management. The aim of this study was to measure basal energy expenditure (BEE), resting energy expenditure (REE) and to explore the true 1 MET value for young Chinese adults with different body weight. METHODS AND STUDY DESIGN: A total of 251 young Chinese adults were divided into three groups: the normal weight group, the overweight group and the obese group. Their BEE, REE and 1 MET values were measured by Cortex Metamax 3B (MM3B). Multiple linear regressions and correlation analysis were used to examine factors that influence EE in Chinese population. RESULTS: The mean measured BEE and REE of young Chinese adults with normal weight, overweight and obesity was 1429, 1609, 1778 kcal/day and 1522, 1712, 1885 kcal/day, respectively. The EE per kilogram body weight decreased with the increasing body weight. The mean oxygen consumption in the three group was 3.78, 3.47, 3.21 mL/kg/min respectively. There was no significant difference in BEE and REE after adjustment for fat-free mass (FFM). The significant influencing factors were body mass index (BMI) and sex in BEE, FFM and BMI in REE in Chinese population. CONCLUSIONS: BEE and REE were significantly different for different body weights while these differences disappeared after adjustment for FFM, and people with different body weights may have different 1 MET values. Further studies should be conducted to obtain more accurate daily energy requirement and 1MET value for specific Chinese populations.

[A cohort study of serum leptin level in Zhejiang Province middle and old age residents on blood pressure and hypertension from 2014 to 2017].

OBJECTIVE: To investigate the association between serum leptin concentration with blood pressure and hypertension in different gender. METHODS: A total of 343 non-hypertensive residents aged 30 to 65 were randomly selected from Zhejiang Province in 2014. Parameters including height, weight, waist, blood pressure, serum lipid and serum leptin concentration of participants were measured and determined by physical examination and laboratory detection in 2014 and 2017. The differences in serum leptin levels between the new hypertensive and non-hypertensive people were compared after three years. The relationship between serum leptin levels and hypertension was analyzed by Logistic regression. RESULTS: In 2014 and 2017, concentration of serum leptin in females was significantly higher than that in males, and the blood pressure in males was higher than that in females. At the end of follow-up, 26 new cases of hypertension were found. Serum leptin levels were higher in the female hypertensive group than in the non-hypertensive group(P=0. 0289), whereas there was no statistical difference in males(P>0. 01). Regardless of gender, serum leptin was significantly positively correlated with body mass index(BMI) and waist-to-height ratio(WHtR)(P<0. 0001), and also had correlation with blood pressure. However, after adjusting for BMI or WHtR, the correlation between serum leptin and blood pressure disappeared(P>0. 01). Logistic regression also showed the similar result. CONCLUSION: Serum leptin is not directly related to blood pressure and hypertension, and the positive correlation between them may be explained largely by BMI and WHtR.

MicroRNA-212 activates hepatic stellate cells and promotes liver fibrosis via targeting SMAD7.

There has been an increasing number of researches about microRNAs (miRNAs) in the progression of liver fibrosis from the point of their comprehensive functions in regulating the activation of hepatic stellate cells (HSCs). Among them, it has been reported that miR-212 is up-regulated in activated rat primary HSCs. However, its mechanism has not been determined yet. Here, we confirmed that the level of miR-212-3p was up-regulated in livers of carbon tetrachloride (CCl4)-treated mice compared with the normal control, which is a classical model of chronically damaged fibrotic liver. In vitro, we demonstrated that TGF-ß, a master fibrogenic cytokine, could induce the level of miR-212. In turn, overexpression of miR-212 could induce the activation marker of HSC including α-smooth muscle actin (α-SMA) and collagens by activating TGF-ß signaling pathway. Furthermore, SMAD7, a dominant suppressor of TGF-ß pathway, was identified as a direct target of miR-212-3p. Our results indicate that miR-212-3p facilitates the activation of HSCs and TGF-ß pathway by targeting SMAD7, highlighting that it can be served as a novel biomarker or therapeutic target for liver fibrosis.