Objective:To investigate effect and mechanism of Morin on inflammatory response of young asthmatic rats by regu-lating mammalian target of rapamycin(mTOR)/signal transducers and activators of transcription 3(STAT3)signaling pathway.Methods:An asthma rat model was established.Experiment was separated into Control group,Model group,Morin low-dose[Morin-L,10 mg/(kg·d)]group,Morin medium dose[Morin-M,30 mg/(kg·d)]group,Morin high-dose[Morin-H,100 mg/(kg·d)]group and Morin high-dose+mTOR activator group[Morin-H+MHY-1485,100 mg/(kg·d)Morin+7 mg/(kg·d)MHY-1485]group.Enhanced expiratory interval value was detected and recorded;Total IgE,ovalbumin(OVA)specific IgE,IL-4,IL-5,IL-17,IL-13,IFN-γ and TGF-β1 levels were determined by ELISA;Giemsa staining was applied to observe and record situation of related inflammatory cells;proportion of Th1,Th2 and Th17 cells were detected by flow cytometry;HE and PAS staining were applied to observe pathologi-cal changes in lung tissue and goblet cell proliferation;GATA-binding protein 3(GATA-3)expression was detected by immunohisto-chemistry and qRT-PCR;Western blot was applied to detect expression and phosphorylation levels of mTOR and STAT3 proteins.Results:Compared with Control group,inflammatory cell infiltration was obvious in Model group,with irregular thickening of tube wall and basement membrane,obviously more goblet cells,and increased mucus secretion,Penh value,IL-4,IL-5,IL-17,IL-13,total IgE and OVA-sIgE levels,macrophages,lymphocytes,eosinophils and neutrophils numbers,Th2 and Th17 cells proportion,average optical density of GATA-3,GATA-3 mRNA and phosphorylation levels of mTOR and STAT3 were obviously increased(P<0.05),proportion of Th1 cells and IFN-γ level were significantly reduced(P<0.05).Compared with Model group,bronchial wall structure of rats in Morin-L,Morin-M and Morin-H groups was smoother and more complete,with epithelial cells arranged in a more orderly manner,moderate airway wall thickness,reduced inflammatory cell infiltration,and reduced goblet cell proliferation,Penh value,IL-4,IL-5,IL-17,IL-13,TGF-β1,total IgE and OVA-sIgE levels,numbers of macrophages,lymphocytes,eosinophils and neutrophils,proportion of Th2 and Th17 cells,average optical density of GATA-3,GATA-3 mRNA,and phosphorylation levels of mTOR and STAT3 were obviously decreased(P<0.05),proportion of Th1 cells and IFN-γ level were significantly increased(P<0.05).MHY-1485 reversed inhibitory effect of morin on inflammatory response in asthmatic rats(P<0.05).Conclusion:Morin may inhibit activation of mTOR/STAT3 signaling pathway,inhibit inflammatory response in young rats with asthma,and thereby improve asthma symptoms.

Stresses induced by the deficiency or excess of trace mineral elements, such as manganese (Mn), represent a common limiting factor for the production of crops like Brassica napus. To identify key genes involved in Mn allocation in B. napus and elucidate the underlying mechanisms, a member of the metal tolerance protein (MTP) family obtained in the previous screening of cDNA library of B. napus under Mn stress was selected as the research subject. Based on the sequence information and phylogenetic analysis, it was named as BnMTP10. It belongs to the Mn-cation diffusion facilitator (CDF) subfamily. Expression of BnMTP10 in yeast significantly improved the tolerance of transformants to excessive Mn and iron (Fe) and reduced the accumulation of Mn and Fe. However, the yeast transformants exhibited no significant changes in tolerance to excess cadmium, boron, aluminum, zinc, or copper. The qRT-PCR results demonstrated that the flowers of B. napus had the highest expression of BnMTP10, followed by roots and leaves. Subcellular localization studies revealed that BnMTP10 was localized in the endoplasmic reticulum (ER). Compared with wild-type plants, transgenic Arabidopsis overexpressing BnMTP10 exhibited enhanced tolerance to excessive Mn stress but showed no significant difference under Fe stress. Correspondingly, under excessive Mn stress, the Mn content in the roots of transgenic Arabidopsis increased significantly. However, under excessive Fe stress, the Fe content in transgenic Arabidopsis did not alter significantly. According to the results, we hypothesize that BnMTP10 may alleviate excessive Mn stress in plants by mediating Mn transport to the ER. This study facilitated our understanding of efficient mineral nutrients, and provided theoretical foundations and gene resources for breeding B. napus.
Brassica napus/genetics* ; Manganese/metabolism* ; Plants, Genetically Modified/genetics* ; Plant Proteins/physiology* ; Arabidopsis/metabolism* ; Gene Expression Regulation, Plant ; Phylogeny ; Cation Transport Proteins/metabolism* ; Stress, Physiological

Objective:To investigate effect and mechanism of Morin on inflammatory response of young asthmatic rats by regu-lating mammalian target of rapamycin(mTOR)/signal transducers and activators of transcription 3(STAT3)signaling pathway.Methods:An asthma rat model was established.Experiment was separated into Control group,Model group,Morin low-dose[Morin-L,10 mg/(kg·d)]group,Morin medium dose[Morin-M,30 mg/(kg·d)]group,Morin high-dose[Morin-H,100 mg/(kg·d)]group and Morin high-dose+mTOR activator group[Morin-H+MHY-1485,100 mg/(kg·d)Morin+7 mg/(kg·d)MHY-1485]group.Enhanced expiratory interval value was detected and recorded;Total IgE,ovalbumin(OVA)specific IgE,IL-4,IL-5,IL-17,IL-13,IFN-γ and TGF-β1 levels were determined by ELISA;Giemsa staining was applied to observe and record situation of related inflammatory cells;proportion of Th1,Th2 and Th17 cells were detected by flow cytometry;HE and PAS staining were applied to observe pathologi-cal changes in lung tissue and goblet cell proliferation;GATA-binding protein 3(GATA-3)expression was detected by immunohisto-chemistry and qRT-PCR;Western blot was applied to detect expression and phosphorylation levels of mTOR and STAT3 proteins.Results:Compared with Control group,inflammatory cell infiltration was obvious in Model group,with irregular thickening of tube wall and basement membrane,obviously more goblet cells,and increased mucus secretion,Penh value,IL-4,IL-5,IL-17,IL-13,total IgE and OVA-sIgE levels,macrophages,lymphocytes,eosinophils and neutrophils numbers,Th2 and Th17 cells proportion,average optical density of GATA-3,GATA-3 mRNA and phosphorylation levels of mTOR and STAT3 were obviously increased(P<0.05),proportion of Th1 cells and IFN-γ level were significantly reduced(P<0.05).Compared with Model group,bronchial wall structure of rats in Morin-L,Morin-M and Morin-H groups was smoother and more complete,with epithelial cells arranged in a more orderly manner,moderate airway wall thickness,reduced inflammatory cell infiltration,and reduced goblet cell proliferation,Penh value,IL-4,IL-5,IL-17,IL-13,TGF-β1,total IgE and OVA-sIgE levels,numbers of macrophages,lymphocytes,eosinophils and neutrophils,proportion of Th2 and Th17 cells,average optical density of GATA-3,GATA-3 mRNA,and phosphorylation levels of mTOR and STAT3 were obviously decreased(P<0.05),proportion of Th1 cells and IFN-γ level were significantly increased(P<0.05).MHY-1485 reversed inhibitory effect of morin on inflammatory response in asthmatic rats(P<0.05).Conclusion:Morin may inhibit activation of mTOR/STAT3 signaling pathway,inhibit inflammatory response in young rats with asthma,and thereby improve asthma symptoms.

Objective To investigate the impact of Atractylodin on lung tissue damage in young asthmatic rats by regulating the CXC chemokine ligand 12(CXCL12)/CXC chemokine receptor 4(CXCR4)signaling pathway.Methods Twelve young SD rats were randomly selected from 60 rats as the control group(CON group),while the remaining 48 rats were used to construct asthma models using ovalbumin(OVA).Successfully modeled asthma rats were randomly separated into Model group,Atractylodin group(50 mg/kg Atractylodin),and CXCL-12 group(5 μ g/kg recombinant CXCL-12 protein)and Atractylodin+CXCL-12 group(50 mg/kg Atractylodin+5 μ g/kg recombinant CXCL-12 protein),with 12 in each group,continuously administered for 14 days.The CON and Model groups were given equal amounts of physiological saline.The percentages of neutrophils and eosinophils in bron-choalveolar lavage fluid(BALF)were detected.ELISA method was applied to detect cytokine levels in serum and BALF fluid;HE staining was applied to detect pathological changes in lung tissue;Western blot was applied to detect the levels of CXCL12/CXCR4 pathway related proteins.Results Compared with the CON group,the pathological score of lung tissue,percentage of neutrophils,percentage of eosinophils,the levels of IL-17,IL-4,IL-5,IL-13,IgE,OVA sIgE,and the protein levels of CXCL12 and CXCR4 in Model group were obviously increased(P<0.05);compared with the Model group,the pathological score of lung tissue,percentage of neutro-phils,percentage of eosinophils,the levels of IL-17,IL-4,IL-5,IL-13,IgE,OVA sIgE,and the protein levels of CXCL12 and CXCR4 in the Atractylodin group were obviously reduced(P<0.05),the results in the CXCL-12 group were opposite to those in the Atractylodes group;CXCL-12 eliminated the improvement effect of atractylodes on asthma rats.Conclusion Atractylodin may improve lung tissue damage in asthmatic rats by down-regulating the CXCL12/CXCR4 signaling pathway.

Objective: To explore the effect of hypoxia inducible factor 1α(HIF-1α)on tamoxifen resistance in breast cancer MCF-7,and to established a human breast cancer cell line(MCF-7/TR)with 4-hydroxy tamoxifen(4-OHT)resistance. Methods: 4-OHT was an active form of tamoxifenin vivo, resistant breast cancer cells MCF-7/TR were establishedin vitrousing 4-OHT. MTT assay was used to detect the effect of 4-OHT on the MCF-7 and MCF-7/TR cells, and the drug resistance multiple was detected. Western blot was used to detect the expression level of HIF-1α protein in MCF-7 and MCF-7/TR cells. The effects of HIF-1α inhibitor(LW6) or siRNA HIF-1α on MCF-7/TR cells and 4-OHT on MCF-7 cells treated with HIF-1α stabilizer(FG-4592) were detected by MTT and flow cytometry AV/PI staining. Results: The results showed that the MCF-7/TR was successfully constructed, and the drug resistance ratio was(5.56±0.80). Compared with MCF-7 cells, MCF-7/TR cells had higher expression of HIF-1α protein and it was up-regulated after tamoxifen treatment. After giving LW6 or silencing the expression of HIF-1α,the down-regulation of HIF-1α expression enhanced the inhibitory effect of 4-OHT on MCF-7/TR cells. After treatment of FG-4592, the expression level of HIF-1α in breast cancer cells MCF-7 was up-regulated, and hindered the inhibition effect of tamoxifen on MCF-7 cells. Conclusion The above results indicate that HIF-1α plays an important role in 4-OHT resistant breast cancer, and targeting HIF-1α may be an effective way to increase the sensitivity of MCF7/TR cells to tamoxifen.

Objective To determine vascular endothelial growth factor(VEGF)-460 gene polymorphism in Uyghurs and its relationship to urolithiasis in south Xinjiang. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP),gene sequencing and genetic analysis methods were used in 200 urolithiasis patients of Uyghurs, and 200 healthy Uyghurs. Results The distribution of genotype and allele had no significant difference between urolithiasis patients and normal controls (P＞0. 05). The frequencies for the CC,TT and CT genotypes in patients with urolithiasis and normal controls were 1.5 %, 29.0 %, 69.5 % and 0. 5 %, 27.5 %, 72.0 %, respectively. The frequencies for C and T allele were 36.2%,63.7% and 36.9% ,63.1%, respectively. Conclusions The results of VEGF-460 gene polymorphisms indicate no significant relationship between patients with turolithiasis and normal controls in Uyghurs in south Xinjiang,which may not be urolithiasis susceptibility genetic locus.