BACKGROUND:miRNAs expression has been reported to be associated with hepatic and renal fibrosis,and dermal fibrogenesis. Moreover,a targeted regulatory relationship between miR-192-5p and epidermal regulators has been demonstrated in gouty arthritis.OBJECTIVE:To investigate the expression and regulatory role of miR-192-5p in hypertrophic scar and to verify whether there is a targeted regulatory relationship between miR-192-5p and epidermal regulators. METHODS:(1) Six cases of hypertrophic scar tissue and six cases of normal skin tissue were collected from the First Affiliated Hospital of Xinjiang Medical University. And miR-192-5p and epidermal regulator mRNA expression were detected by qRT-PCR. (2) The primary hypertrophic scar fibroblasts were obtained using tissue explant method and cultured to 3-6 generations for subsequent experiments. There were three groups in the experiment:negative control group,miR-192-5p mimic group and miR-192-5p inhibitor group. The latter two groups were transfected with the corresponding sequences. Cell proliferation viability was detected by the cell counting kit-8 assay and EdU kit;and the migration ability was detected by the cell scratch test. Cell apoptosis was detected by flow cytometry. The gene and protein expressions of epidermal regulator,type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were detected by qRT-PCR and western blot,respectively. miR-192-5p targets were predicted by a bioinformatics website,and target binding was validated by dual luciferase assay. RESULTS AND CONCLUSION:(1) Compared with normal skin tissues and their fibroblasts,miR-192-5p and epidermal regulator were highly expressed in hypertrophic scar and hypertrophic scar fibroblasts (P<0.05 or P<0.01). (2) After overexpression of miR-192-5p,cell proliferation was enhanced (P<0.05) and EdU positive cell rate increased (P<0.01) when compared with the negative control group;after inhibition of miR-192-5p,cell viability (P<0.05) and EdU positive rate decreased (P<0.05). (3) At 24 hours after overexpression of miR-192-5p,compared with the negative control group,the area between cell scratches and apoptosis rate decreased in the miR-192-5p mimic group (P<0.05) but increased in the miR-192-5p inhibitor group (P<0.01). (4) At 48 hours after transfection,the mRNA and protein levels of epidermal regulator were significantly decreased in the miR-192-5p mimic group,while the mRNA and protein levels of type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were significantly increased (P<0.05 or P<0.01). The miR-192-5p inhibitor group showed opposite changes in the above four indicators (P<0.05 or P<0.01). (5) The Targetscan website predicted that epidermal regulator had a potential binding site for miR-192-5p. (6) Dual luciferase assays showed that miR-192-5p could bind to epidermal regulator in a targeted manner. To conclude,overexpression of miR-192-5p can decrease the expression of epidermal regulator,and the two may be negatively regulated,suggesting that regulation of epidermal regulator may play a role in inhibiting the proliferation of hypertrophic scar fibroblasts.

BACKGROUND:miRNAs expression has been reported to be associated with hepatic and renal fibrosis,and dermal fibrogenesis. Moreover,a targeted regulatory relationship between miR-192-5p and epidermal regulators has been demonstrated in gouty arthritis.OBJECTIVE:To investigate the expression and regulatory role of miR-192-5p in hypertrophic scar and to verify whether there is a targeted regulatory relationship between miR-192-5p and epidermal regulators. METHODS:(1) Six cases of hypertrophic scar tissue and six cases of normal skin tissue were collected from the First Affiliated Hospital of Xinjiang Medical University. And miR-192-5p and epidermal regulator mRNA expression were detected by qRT-PCR. (2) The primary hypertrophic scar fibroblasts were obtained using tissue explant method and cultured to 3-6 generations for subsequent experiments. There were three groups in the experiment:negative control group,miR-192-5p mimic group and miR-192-5p inhibitor group. The latter two groups were transfected with the corresponding sequences. Cell proliferation viability was detected by the cell counting kit-8 assay and EdU kit;and the migration ability was detected by the cell scratch test. Cell apoptosis was detected by flow cytometry. The gene and protein expressions of epidermal regulator,type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were detected by qRT-PCR and western blot,respectively. miR-192-5p targets were predicted by a bioinformatics website,and target binding was validated by dual luciferase assay. RESULTS AND CONCLUSION:(1) Compared with normal skin tissues and their fibroblasts,miR-192-5p and epidermal regulator were highly expressed in hypertrophic scar and hypertrophic scar fibroblasts (P<0.05 or P<0.01). (2) After overexpression of miR-192-5p,cell proliferation was enhanced (P<0.05) and EdU positive cell rate increased (P<0.01) when compared with the negative control group;after inhibition of miR-192-5p,cell viability (P<0.05) and EdU positive rate decreased (P<0.05). (3) At 24 hours after overexpression of miR-192-5p,compared with the negative control group,the area between cell scratches and apoptosis rate decreased in the miR-192-5p mimic group (P<0.05) but increased in the miR-192-5p inhibitor group (P<0.01). (4) At 48 hours after transfection,the mRNA and protein levels of epidermal regulator were significantly decreased in the miR-192-5p mimic group,while the mRNA and protein levels of type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were significantly increased (P<0.05 or P<0.01). The miR-192-5p inhibitor group showed opposite changes in the above four indicators (P<0.05 or P<0.01). (5) The Targetscan website predicted that epidermal regulator had a potential binding site for miR-192-5p. (6) Dual luciferase assays showed that miR-192-5p could bind to epidermal regulator in a targeted manner. To conclude,overexpression of miR-192-5p can decrease the expression of epidermal regulator,and the two may be negatively regulated,suggesting that regulation of epidermal regulator may play a role in inhibiting the proliferation of hypertrophic scar fibroblasts.

Objective: To investigate the genetic characteristics of human norovirus (NoV) among infants under 5 years of age with diarrhea in Chaoyang District, Beijing from 2011 to 2017. Methods: NoV-positive stool samples were collected from 2011 to 2017 in this region. The partial RdRp and VP1 genes were amplified and sequenced. Multi-sequence alignment was performed and phylogenetic tree was constructed using Mega software. Results: A total of 151 samples were sequenced and analyzed. The ratio of male and female was 2.28∶1 with mean age of 1.72 years. Fourteen NoV subtypes were detected, including GII.Pe/GII.4 (47.68%), GII.P12/GII.3 (20.53%), GII.P4/GII.4 (17.22%), GII.P16/GII.2 (3.31%), GII.P12/GII.12 (1.99%), GII.P17/GII.17 (1.99%), GII.P16/GII.13 (1.32%), GII.P7/GII.7 (1.32%), GII.P7/GII.6 (1.32%), GII.P2/GII.2 (0.66%), GII.P21/GII.21 (0.66%), GII.Pg/GII.12 (0.66%), GI.Pa/GI.3 (0.66%) and GI.P6/GI.6 (0.66%). Conclusions NoV genetic diversity was found among infants under 5 with diarrhea in Chaoyang district, Beijing. The subtypes from surveillance and those from epidemics occurred in chronological order. The surveillance should be strengthened for early detection of new subtype for monitoring the epidemic and vaccine design.

Objective: To investigate the genetic characteristics of human adenovirus (AdV) among infants with diarrhea in Chaoyang district, Beijing from 2011 to 2017. Methods: Adenovirus positive stool samples were collected from 2011 to 2017 in Chaoyang District of Beijing. The hexon region genes of human adenovirus were sequenced. Multi-sequence alignments were performed and phylogenetic tree was constructed by Mega software. Results: A total of 64 samples were sequenced and analyzed. The ratio of male to female was 11∶5. The mean age was 1.56 years. Among them, AdV41 accounted for 70.31%, followed by AdV31 (26.25%), AdV40 (4.69%), AdV1 (3.13%), AdV5 (3.13%), AdV6 (3.13%), AdV7 (3.13%), AdV2 (1.56%), AdV3 (1.56%), AdV4 (1.56%) and AdV61 (1.56%). Conclusions Human adenovirus may play an important role in viral diarrhea in Chaoyang district from 2011 to 2017. The current adenovirus epidemic is complex and AdV41 was the dominant strain in this region.

Objective: To investigate the level of interleukin-6 (IL-6) and the effect of valproic acid(VPA) administration on IL-6 promoter methylation, further to explore the epigenetic mechanism in febrile seizures. Methods: Sprague-Dawley (SD) rats (21 day) were randomly divided into control group (n=12) and febrile seizure (FS) group (n=12), and seizures were generated using hot bath methods and evaluated by racine score and electroencephalogram (EEG). The IL-6 protein and mRNA levels were detected using ELISA and quantitative reverse transcription polymerase chain reaction (qRT-PCR), respectively.Rat C6 cell line was cultured and randomly divided into control, VPA(0.2 mol/L), hyperthermia(43.5 ℃, 1 h), hyperthermia (43.5 ℃, 1 h)+VPA (0.2 mol/L) groups.The methylation status of IL-6 promoter in FS rats and the effect of VPA on IL-6 methylation in C6 cell line were examined by bisulfite sequencing polymerase chain reaction (BSP) to determine the epigenetic regulation of IL-6 level in FS. Results: Racine score and EEG results showed that the frequency and amplitude of neuronal discharge increased after hot water bath in rats, and the latency of convulsion was shorter.The expression levels of IL-6 mRNA and protein were significantly increased in Fs group compared with those in control group(mRNA: Control: 0.98±0.34; FS: 2.85±0.39, P<0.05; Protein: Control: (2 824.33±169.20)pg/ml; FS: (3 514.58±86.4)pg/ml, P<0.01). The methylation of IL-6 promoter in FS group (84% (21/25)) was lower than that in control group (100% (25/25)) (P<0.05). A significant increase in methylation of IL-6 promoter was observed in C6 cell line from VPA combined with hyperthermia (96% (24/25)) but no change was showed in that from VPA alone (100% (25/25)) compared with that from control(100% (25/25)). Conclusion IL-6 level is up-regulated in FS rats, and the hypomethylation or demethylation of the IL-6 promoter region might increase the IL-6 level.VPA administration can elevate its methylation level, which provides a strong experimental basis for the future study of the epigenetic mechanism in FS.

BACKGROUND: Establishment of lumbar disc degeneration in animal models can help doctors understand the development rules and pathophysiological changes of this disease, and can explore and research more rational treatment by creating animal models.OBJECTIVE: To review recent advances in vivo model of lumbar disc degeneration.METHODS: The first author used the computer to search related articles on PubMed database and Chinese Journal Full-text Database from inception to May 2016. The search key words were lumbar disc, degeneration, animal model,vivo model. A total of 161 related articles were retrieved and 49 of them met the inclusion criteria.RESULTS AND CONCLUSION: (1) Building an intuitive and reliable animal model of lumbar disc degeneration will not only help the basic research of degenerative disc disease, but also provide a good experimental carrier for the repair treatment of degenerative lumbar disc. (2) After more than 80 years of development, the animal model of lumbar disc degeneration has formed a production mode of more species, more methods and newer ideas. Current models have their own advantages, cannot be replaced. However, problems such as poor controllability, low degree of safety or long observation period are still exiting more or less. All these need further research and exploration.

BACKGROUND:Differentiation of bone marrow mesenchymal stem cels is induced by integrated factors.In vitro interaction of cytokine complex and certain cel mechanical stimulation is carried out to further improve the efficiency of bone marrow mesenchymal stem cels differentiating into nucleus pulposus-like cels. OBJECTIVE:To investigate the differentiation of bone marrow mesenchymal stem cels into nucleus pulposus-like cels induced by transforming growth factor-β1 and insulin-like growth factor-1 under hydrostatic pressure. METHODS: Bone marrow mesenchymal stem cels from adult rats were separated, cultured and purified in vitro. Passage 3 cels were induced in vitrowith transforming growth factor-β1 and insulin-like growth factor-1 under hydrostatic pressure (hydrostatic pressure group), with transforming growth factor-β1 and insulin-like growth factor-1 under normal pressure (drug group), or with normal culture medium under normal pressure (blank control group). RESULTS AND CONCLUSION:At day 14 after culture, polygonal nucleus pulposus-like cels were observed in the hydrostatic pressure group, but irregular cels in the drug group. There was no obvious change in the blank control group. Levels of colagen type II and DNA were higher in the hydrostatic pressure group than the other two groups. These findings indicate that the combination of transforming growth factor-β1 and insulin-like growth factor-1 can successfuly induce the differentiation of bone marrow mesenchymal stem cels into nucleus pulposus-like cels under hydrostatic pressure, and the differentiation efficiency is higher under hydrostatic pressure than under normal pressure.

Objective To investigate the effect of haplotype and linkage disequilibrium of PPARγgene rs3856806, rs12490265, rs1797912, and rs1175543 in patients with metabolic syndrome ( MS) in Kazakhs of Xinjiang.Methods MALDI-TOF-MS was used to detect PPARγgene rs3856806, rs12490265, rs1797912, and rs1175543 genotypes in 489 subjects ( including 245 MS and 244 controls ) .Results ( 1 ) The frequencies of rs3856806T, rs12490265A, rs1797912C and rs1175543G alleles for MS group in Kazakhs were all significantly lower than those for controls [ rs3856806T allele:12.53% vs 17.01%; rs12490265A allele: 31.84% vs 38.52%;rs1797912C allele:35.31%vs 43.24%;rs1175543G allele:40.61%vs 47.54%(all P<0.05)].(2)Significant linkage disequilibrium were observed between PPARγgene rs1797912 and rs1175543, rs12490265, and rs1175543 polymorphisms.(3)AGCC and GAAT were significantly different between MS and control group in Kazakhs(both P<0.05).(4) Carrying rs3856806T, rs12490265A, rs1797912C, rs1175543G was 0.267 times that of carrying rs3856806C, rs12490265G, rs1797912A, rs1175543A.Conclusions The PPARγgene rs3856806, rs12490265, rs1797912 and rs1175543 polymorphisms were associated with metabolic syndrome in Kazakhs.There were very strong linkage disequilibrium between PPARγgene rs1797912 and rs1175543, rs12490265 and rs1175543 polymorphisms, The AGCC haplotype and GAAT haplotype may serve as protective factors of metabolic syndrome.Carrying rs3856806T, rs12490265A, rs1797912C, and rs1175543G may confer lower risk of MS in Kazakhs.