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Objective:To investigate the possible role of long non-coding RNA (LncRNA) 00602 in promoting browning in adipocytes induced by adenovirus type 36 (Ad36).Methods:According to Ad36 infection, adipose tissue samples of obese patients were divided into Ad36-negative group and Ad36-infected group. Realtime fluorescent quantitative PCR (qRT-PCR) was used to detect the changes in the expression of LncRNA00602 mRNA in omental adipose tissue of the two groups, and analyze the differences between the two groups. The correlation between waist-to-hip ratio, systolic blood pressure, diastolic blood pressure, fasting blood glucose, triacylglyceride and other indicators of the patients in the group with LncRNA00602 mRNA expression were analyzed. HE staining was used to detect the size of adipocytes in the omental adipose tissue of the Ad36 negative group and the Ad36 infection group. qRT-PCR and Western blotting were used to detect the mRNA and protein expression levels of uncoupling protein 1 (UCP1) and PR domain containing 16 (PRDM16) in omental adipose tissue of two groups of patients. Human adipose-derived stem cells (hADSC) were isolated and cultured, using Ad36 to induce differentiation, and divided into control group and LncRNA00602 knockdown group. On 0, 2, and 4 days after LncRNA00602 knockdown, fluoroboron dipyrrole (BODIPY) and mitochondrial red fluorescence (Mito-Tracker Red) were used to stain intracellular lipid droplets and mitochondria. At the same time, qRT-PCR and Western blotting were used to detect changes in the expression of UCP1 and PRDM16.Results:The expression of LncRNA00602 gene in the Ad36 infection group was higher than that in the Ad36 negative group (all P<0.05). The expression of LncRNA00602 in the Ad36 negative group was not significantly different from the above clinical indicators, while the expression of LncRNA00602 was negatively correlated with serum fasting blood glucose and triacylglyceride ( r=-0.522, -0.486, P<0.05) in the Ad36 infection group; HE staining showed that the average adipocyte area of the Ad36 infection group was smaller than that of the Ad36 negative group. At the same time, UCP1 and PRDM16 gene expression were higher than the negative group (all P<0.05). At the cellular level, on the 2nd and 4th days after knockdown of LncRNA00602, the lipid droplet area of adipocytes in the LncRNA00602 knockdown group was larger than that of the control group, the number of mitochondria decreased compared with the control group, and difference was statistically significant ( P<0.05 or P<0.01); Compared with the control group, there was significantly lower expression of the browning marker genes UCP1, PRDM16, and protein in the adipocytes in the LncRNA00602 knockdown group (all P<0.05). Conclusion:In Ad36-induced adipocyte differentiation, LncRNA00602 may positively regulate the expression of UCP1, PRDM16 and lipid droplet metabolism, and promote the browning of adipocytes.
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Objective@#To investigate the role of adenovirus type 36 (Ad36) in the browning of 3T3-L1 cells.@*Methods@#BODIPY staining was performed on the 0, 2nd, 4th, 6th and 8th days of the cocktail induction (control) group and the cocktail plus Ad36 induction (experimental) group to observe the adipogenesis of 3T3-L1 cells.The mRNA and protein expressions of uncoupling protein-1(Ucp1), ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit (Atp5o), cytochrome c oxidase subunit 5B(Cox5b), and perilipin were detected by real-time PCR and Western-blot.@*Results@#The results of BODIPY staining showed that the lipid droplets in the control group gradually became larger with the differentiation of the cells, while the lipid droplets in the experimental group firstly became larger, and then appeared smaller after Ad36 was added on the fourth day. Compared with the control group, the mRNA and protein expression levels of Ucp1, Atp5o, and Cox5b in the experimental group were significantly increased while the expression level of perilipin was significantly decreased (all P<0.05).@*Conclusion@#During the differentiation of 3T3-L1 cells, Ad36 promotes its browning via increasing the expressions of Ucp1, Atp5o, and Cox5b.
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Objective To investigate the role of adenovirus type 36 ( Ad36) in the browning of 3T3-L1 cells. Methods BODIPY staining was performed on the 0, 2nd, 4th, 6th and 8th days of the cocktail induction (control) group and the cocktail plus Ad36 induction ( experimental) group to observe the adipogenesis of 3T3-L1 cells. The mRNA and protein expressions of uncoupling protein-1 ( Ucp1 ), ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit ( Atp5o), cytochrome c oxidase subunit 5B ( Cox5b), and perilipin were detected by real-time PCR and Western-blot. Results The results of BODIPY staining showed that the lipid droplets in the control group gradually became larger with the differentiation of the cells, while the lipid droplets in the experimental group firstly became larger, and then appeared smaller after Ad36 was added on the fourth day. Compared with the control group, the mRNA and protein expression levels of Ucp1, Atp5o, and Cox5b in the experimental group were significantly increased while the expression level of perilipin was significantly decreased ( all P＜0. 05 ). Conclusion During the differentiation of 3T3-L1 cells, Ad36 promotes its browning via increasing the expressions of Ucp1, Atp5o, and Cox5b.
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Objective To investigate the role of LncRNA ROR in Ad36-induced browning of human adipose-derived stem cells(hADSC). Methods After hADSC was induced by cocktail and Ad36 for 2,4,6,and 8 days,Oil red O staining was performed for observing the adipogenic status. The mRNA expressions of LncRNA ROR, uncoupling protein 1(UCP1),and PRDM16 were detected by real-time PCR and the protein expressions of UCP1 and PRDM16 were detected by Western-blot. After LncRNA ROR was knocked down by siRNA, UCP1 and PRDM16 mRNA and protein expression levels in the process of Ad36-induced adipocyte differentiation were detected by real-time PCR and Western-blot. Results Oil red O staining showed that fat droplets in the cocktail-induced group were larger than those in the Ad36-induced group. Compared with the cocktail group,the mRNA expressions of LncRNA ROR, UCP1 and PRDM16, and the protein expressions of UCP1 and PRDM16 in Ad36 group were significantly increased(P<0.05). The mRNA and protein expressions of UCP1 and PRDM16 in LncRNA ROR knockdown group were significantly lower than those in control group(P<0.05). Conclusion In the process of Ad36-induced hADSC differentiation,the up-regulation of LncRNA ROR may stimulate UCP1 and PRDM16 expression,and thus promote the browning of hADSC.
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Objective To investigate the expressions of RNA-binding protein human antigen R(HuR), fatty acid binding protein type 4(FABP4),fatty acid synthetase(FASN),and lipoprotein lipase(LPL)during the differentiation of human adipocytes, and to explore their possible roles. Methods Human adipose-derived mesenchymal stem cells were induced by adipogenic differentiation,and the adipogenesis of cells was observed by oil red O staining. The expressions of HuR,FABP4,FASN,and LPL mRNA and protein were detected by real-time PCR and Western blotting. After HuR was silenced by siRNA, the change of adipogenesis for human adipose-derived mesenchymal stem cells was observed and the expressions of adipogenic genes were detected. Results The expressions of HuR,FABP4,FASN,and LPL mRNA and protein were significantly increased after human adipose-derived mesenchymal stem cells were induced to differentiate into adipocytes(all P<0.01). After HuR expression was down-regulated by siRNA,the adipogenic level of human adipose-derived mesenchymal stem cells was reduced,with decreased protein levels of FABP4,FASN,and LPL(all P<0.05),which were without changes for their mRNA levels. Conclusion HuR promotes the differentiation of human adipocytes mainly via regulating the changes of FABP4,FASN,and LPL protein levels.
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Objective To investigate the effectiveness of problem-based learning in the biochemistry teaching in merging class of minority and Han students.Methods Totally 460 clinical medical students were divided into PBL group which contained 252 students and the traditional teaching group which involved 208 students,respectively.According to each team of seven to eight students,minority and Han students randomly arranged.Control group used classroom teaching mode,experimental group in addition to classroom lectures,had additional 12 hours of PBL teaching,but the theory classes for the two groups of students were taught by the same six teachers with rich teaching experience,and the teaching content and teaching material selection were also the same.At the end of the course,the learning outcomes were evaluated by using descriptive analysis and t test (α=0.05) based on the combination of theoretical examination,experimental practice and the questionnaire survey method.Results Compared with the traditional teaching group,the final scores were higher than those of PBL group (84.72 ± 6.99 and 80.34 ± 7.12,P＜0.05).There were also statistically significant between two groups according to different nationality(Minority:85.65 ± 5.27 and 79.70 ± 7.14;Han:83.91 ± 8.26 and 80.95 ± 7.08;P＜0.05),and interestingly the increased ratio of scores was higher in minority than that in Han.The questionnaire surveys indicated that the PBL teaching method could enhance professional and comprehensive qualities of students and more than 81.83％ students were satisfied with the new teaching mode.Conclusions The combination of tradition and PBL-based teaching methods improved the quality of biochemistry teaching of clinical medicine in merging class of minority and Han students in Xinjiang Medical University.
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ABSTRACT:Objective To investigate the regulation of adenovirus type 36 infection on precursor protein particles progranulin expression in Uygur obese patients.Methods Based on the diagnostic criteria of obesity,the samples were divided into obese group and non-obese group.Serum neutralization test was used to detect the antibody of Ad36.The progranulin mRNA expressions in abdominal omental and subcutaneous adipose tissues were detected by real-time quantitative PCR.ELISA method was used to determine serum progranulin protein levels. CD68 protein expression of macrophage was detected by immunohistochemistry. Results ① In the Uygur population of our study,compared with that in the non-obese group (38/1 1 7,32.5%),the number of obese patients (54/1 1 5,47.0%)infected with Ad36 was significantly higher than that in non-obese group (P <0.05 ).② Serum progranulin was significantly increased in the Ad36-infected obese group (408.45±1 56.92)than in non-obese group (326.1 1±1 58.60)(P <0.05).The mRNA expression of progranulin did not differ between the two groups.③ The macrophage infiltration was significantly higher in the Ad36-infected obese group (14 730.1 6 ± 2 227.39 )than in non-obese group (10 786.50 ± 2 772.80 )(P < 0.05 ).Conclusion Ad36 infection may be associated with the occurrence of obesity in Uygur population,and adenovirus type 36 infection may regulate the expression of serum progranulin at the protein level.
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0.05).The expression of survivin mRNA and BCRP mRNA was correlated with therapeutic effect of chemotherapy(P