Objective To explore the status of C57BL/6J mouse brown fat adipogenic differentiation function with aging.Methods C57BL/6J female and male mice at the ages of 0-week (newborn),4-week,8-week,12-week old were selected from the same brood,brown adipose tissue was obstained from their interscapular region,and the brown adipose was identified by using immunohistochemical markers.Then the total RNA was extracted from the brown adipose and quality identification was determined at the same time.The expression levels of the related genes (PPARα,C/EBPα,PGC-1α,PPARγ,FOXC2,BMP7) induced by brown adipose adipogenic differentiation were detected by quantitative real-time PCR in 0-week,4-week,8-week,12-week mice.Results Uncoupling protein -1 (UCP1) immunohistochemical data indicated that positive deep-colour substance was brown adipose tissue.Quantitative Real-time PCR also indicated that the expression volume of adipogenesis gene gradually reduced with aging,and there were significant differences at the different time points [PPARα (F =11.96,P ＜ 0.000 1),C/EBPα (F =9.39,P ＜0.000 1),PGC-1α(F =17.21,P ＜0.000 1),PPARγ(F =13.11,P ＜0.000 1),FOXC2(F =12.23,P ＜0.000 1),BMP7(F =16.44,P ＜0.000 1)].Conclusions The adipogenic differentiation ability and activity of mouse brown adipose gradually reduce with aging.But the regulatory factors for its function needs to be further investigated.

Objective To establish an effective DNA isolation method for neonatal disease screening,so as to explore its application to the methylation detection.Methods The 20 dried blood spots samples were randomly divided into 2 groups according to the gender:the traditional method group (n =10) and the improved kit method group(n =10).The DNA quality was evaluated based on its concentration,integrity and whether it could be used in polymerase chain reaction (PCR).These DNA samples with or without bisulfite treatment were used as template in the methylation-specific polymerase chain reaction (MSP).The methylation levels of Leptin and tumor necrosis factor-α (TNF-α) gene promoter region were detected.Results DNA concentration of the improved kit method [(5.70 ± 0.81) mg/L] was significantly higher than that of the traditional method [(3.50 ± 0.45) mg/L] (t =2.79,P ＜ 0.05),and biochemical analyzer analysis showed a better DNA integrity.Agarose gel electrophoresis revealed that 18S gene fragment could be successfully amplified by PCR method,suggesting its potential application to PCR study.MSP results showed different DNA methylation levels of Leptin and TNF-α genes promoter regions from various samples.Conclusions The improved kit method can effectively extract DNA from dried blood spots samples,and these DNA can be used in methylation research.The study can provide a new research direction and technical method to reveal the pathogenesis of disease from the perspective of DNA methylation.

Objective To investigate the inlfuence of overexpression of miR-26b on the secretion of adipokines dur-ing human adipocyte differentiation. Methods Human preadipocytes were infected with the hsa-miR-26b over-expressing lentivirus and were induced to differentiate, and then the levels of adipokines (IL-6, leptin, resistin, TNF-α) at different time points during differentiation were measured by ELISA. Results Compared with control group, decreased secretions of both IL-6 and leptin, and increased secretion of resistin were found during the differentiation of human adipocytes in miR-26b overexpressed group. However, the secretion of TNF-αwas not measured in both groups. Conclusion The miR-26b can improve the inlfammation and insulin resistance of human adipocytes, which will provide potential targets for obesity treat-ment.

Objective To predict the biological process and signaling pathways in which hsa-miR-1908 might be in-volved by a series of bioinformatics analysis, so as to lay foundations and provide theoretical basis for the further studies of hsa-miR-1908 biological function in human preadipocytes. Methods The sequence of hsa-miR-1908 was acquired from miR-Base database, and target genes of hsa-miR-1908 were predicted by miRanda, and then the intersection of the results and the results of gene-chip as gene set were further analyzed by gene ontology and pathway enrichment. Results The hsa-miR-1908 had some conserved property among different species. The functions of the target genes were enriched in Wnt receptor signal-ing pathway through beta-catenin, cell cycle, cell apeptosis and other biological processes. The GnRH signaling, MAPK sig-naling, insulin signaling, cell cycle signal transduction pathways and signaling pathway in pancreatic cancer were signiifcantly enriched. Conclusions The target genes set of hsa-miR-1908 were enriched in multiple biological process which are related with the obesity. This study provides guidance for the further study in human preadipocytes.

Objective To observe the effect of Triclosan( TCS) exposure on Caenorhabditis elegans( c. ele-gans) F1 generation of locomotory behavior, brood size, and generation time. Methods The trial included a control group and 4 TCS treatment groups with different doses (100 nmol/L,1 μmol/L,10μmol/L,20μmol/L),the exposure time being 24 hours,the effect of c. elegans′head thrashes,body bending frequency,the brood size and generation time was observed. Results (1) The control group exposed to 100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,their head thrash frequency of c. elegans F1 was(109. 40±8. 61) times/min,(84. 70±7. 82) times/min,(76. 35±7. 44) times/min,(74. 74±5. 93)times/min,(71. 95±4. 19)times/min,respectively,the head thrash ability of c. elegans was significantly inhibited(F=62. 245,P<0. 01). (2) When the control group was exposed to 100 nmol/L,1 μmol/L,10μmol/L,20 μmol/LTCS,the frequency of c.elegans F1 body bent was (19.94±2.46)times/20 s,(15.13±1.99) times/20 s,(14.63±2.31)times/20 s,(14.69±1.96)times/20 s,(12.00±1.86)times/20 s,respectively,and the comparative differences between groups were statistically significant(F=25. 636,P<0. 01). (3) When the control group was exposed to 0,100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,the body sizes of the c. elegans F1 generation was (286.83±6.01)articles,(273.33±6.41)articles,(214.17±7.25)articles,(173.67±9.20)articles, (118. 50 ± 6. 98 ) articles, respectively, the brood size of the C. elegans F1 generation exposed to 100 nmol/L, 1μmol/L,10 μmol/L,20 μmol/L TCS levels,were reduced by 4. 71%,25. 60%,39. 45%,58. 67%,the ge-neration time of the c. elegans′F1 generation was shortened by 2. 14%-5. 38% in the TCS treatment groups compared with the control group(F=27. 520,P<0. 01). Conclusions After c. elegans exposure to TCS,locomotory behavior can be severe-ly affected,reproductive damage causes a decline in the number of brood size,and the speeding-up of the breeding rate is related to the concentration of TCS concentration-response.

Objective To predict the functions of hsa-miR-1908 promoter using various bioinformatic tools, and to provide clues for further study on transcriptional regulation mechanism of miR-1908 in human adipocytes. Methods The promoter se-quence of miR-1908 was obtained from Ensemble, and then the CpG islands and transcription factor binding sites were pre-dicted by a variety of online bioinformatic tools. Results The length of the miR-1908 promoter sequence was 1 458 bp. The CpG islands, which inhibited the transcription of miR-1908, were located at (438-756) bp, (836-937) bp and (979-1374) bp. Meanwhile, 15 transcription factor binding sites were found in the promoter sequence of miR-1908. Conclusions miRNA up-stream promoter related bioinformatics can not only improve the efficiency of microRNA promoter research, but also provide further important information on transcriptional regulation of miR-1908.

Necroptosis is a regulated process of programmed cell death independent of aspartic acid-specific cysteine protease,which can induce inflammation.Studies have shown that necroptosis is closely related to the progression and prognosis of pancreatic disease and plays an important two-way regulatory role in its progression.Related necroptosis inhibitors and inducers are expected to be used in the treatment of pancreatic disease.We herein review the mechanism of necroptosis and its role in the progression of pancreatic disease to provide a new understanding of the pathogenesis and treatment of pancreatic diseases and offer a theoretical basis for the research and development of targeted drugs.