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Objective To explore the effect and mechanism of palmitic acid sodium(PA) exposure for different time on HK-2 cells injury. Methods CCK-8 assay was used to detect the activity of HK-2 cells treated with PA at different times; oil red staining was used to detect lipid deposition at different time of PA treatment; Annexin-Ⅴ-FITC/PI apoptosis kit was used to detect the apoptosis of HK-2 cells treated with PA at different times; reactive oxygen species(ROS) production was detected by ROS kit at different time of PA treatment; ELISA was used to detect GSH and SOD in cells as well as the levels of IL-1β and IL-18 in the cell supernatant; Western blot was used to determine the expressions of NLRP3, ASC, Caspase-1 and NOX4 in HK-2 cells. Results CCK-8 assay showed that HK-2 cell viability decreased in PA(6, 9, 12, 24 h) groups; oil red staining showed that lipid deposition increased in PA(6, 9, 12, 24 h) groups; Annexin-Ⅴ-FITC/PI apoptosis kit results showed that the apoptosis rate increased; ROS kit results showed that ROS production significantly increased in PA(6, 9, 12, 24 h) groups; ELISA results showed that the activities of GSH and SOD decreased in PA(6, 9, 12, 24 h) groups, while the levels of IL-1β and IL-18 in supernatant increased; Western blot results showed that the expression levels of inflammatory related proteins NLRP3, Caspase-1, ASC and NOX4 in PA(6, 9, 12 and 24 h) increased. Conclusion PA exposure can induce lipid deposition and HK-2 cells damage, and the mechanism may be related to PA-induced inflammatory response by activating NLRP3 inflammasome in HK-2 cells.

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Objective To investigate the role and mechanism of macrophage-to-myofibroblast transition(MMT) induced by exosomes from high glucose-treated renal tubular epithelial cells. Methods Human renal tubular epithelial cells(HK2) were divided into glucose control group(5.5 mmol/L D-glucose) and high glucose group(30.0 mmol/L D-glucose) and cultured for 48 hours. The supernatant was collected by ultracentrifugation and identified by transmission electron microscopy and Western blot. PKH67 labelled exosomes were used to stimulate THP-1 macrophages. Laser scanning confocal microscopy was used to observe the phagocytosis process. The expression of inducible nitric oxide synthase(iNOS), α-SMA and CD206 was detected by flow cytometry and Western blot to determine the best concentration and time point. Laser scanning confocal microscopy was used to detect the fluorescence co-expression of rabbit polyclonal to mannose receptor(CD206), α-smooth muscle actin(α-SMA), collage type-Ⅳ(Col-IV) and fibronectin(FN). The expression of tumor growth factor-beta 1(TGF-β1), interleukin(IL)-6,IL-10 was separately detected by real-time quantitative PCR(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA). The expression of TGF-β1, mothers against decapentaplegic homolog 3(Smad3), phosphorylated-smad 3(p-Smad3) in THP-1 macrophages was detected by Western blot. Results The expression of cluster of differentiation 63(CD63)and tumor susceptibility gene 101 protein(TSG101)was positive, while the Calnexin protein was negative in the supernatant, confirming that the specimen was exosomes with high purity. THP-1 macrophages could internalize each group of exosomes. 40 mg/L exosomes for 96 hours were the best experimental condition. After being stimulated by high-glucose-exosomes, the percentage of M1 macrophages would climax in 24 hours, and the rate of M2 macrophages would climax in 96 hours. Immunofluorescence staining showed that exosomes released by HG-treated HK2 induced CD206, α-SMA, Col-IV and FN accumulation in cultured THP-1 macrophages. Compared with normal-glucose-exosomes, high-glucose-exosomes increased the expression of TGF-β1, IL-10 in M2 macrophages and decreased the expression of IL-6(allP<0.05). Moreover, TGF-β1 and p-Smad3 proteins expression also increased significantly(allP<0.05). Conclusion Exosomes secreted by renal tubular epithelial cells in high glucose environment can induce M2 macrophages transdifferentiate into myofibroblasts, and its mechanism may be related to activation of TGF-β1/Smad3 signal pathway.

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Objective To investigate the effect of capsaicin on retinal vascular permeability in diabetic rats(DM). Methods 24 eight-week-old male sprague-dawley(SD) rats were randomly categorized into four groups(n=6): control group(NC), diabetic group(DM), diabetic group with 3 mg/kg capsaicin treatment(CAP3) and diabetic group with 6 mg/kg capsaicin treatment(CAP6). Type 2 diabetic rat(T2 DM) model was established by high-fat diet combined with small-dose streptozotocin(STZ)induction. The body weight and fasting blood glucose in 4 groups were monitored. Total cholesterol(TC), triglycerides(TG), low density lipoprotein(LDL), high density lipoprotein(HDL) concentrations in serum were detected according to manufacturer′s instructions. Evans blue angiography was used to observe the changes of retinal vascular permeability.Immunofluorescence was used to measure the distribution and expression of transient receptor potential vanillic 1(TRPV1) in retina. Western blot was used to detect the expression of TRPV1, vascular endothelial growth factor(VEGF) and occludin in rat retina. Results Compared with the DM group, serum total cholesterol(TC), triglyceride(TG) and low density lipoprotein(LDL) levels in the CAP3 and CAP6 groups significantly decreased(P<0.01). The results of angiography showed that the retinal vascular permeability increased in the DM group and that there were many strong fluorescent leakage areas, while the leakage was reduced in CAP3 and CAP6 groups. In CAP3 and CAP6 groups, immunofluorescence showed the activation of TRPV1 in the retinal ganglion cell layer, inner nuclear layer and outer nuclear layer. Western blot indicated that the expression of TRPV1 and occludin increased while the expression of VEGF decreased(P<0.01). Conclusion Capsaicin may exert the effect of improving diabetic retinopathy, underlying which the mechanism may be that capscicin activates retinal TRPV1, resulting in the downregulation of VEGF and upregulation of occuldin.

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Objective To analyze the changes of intestinal flora and explore the possible mechanisms of alcoholic liver injury in mice. Methods Six-week-old mice were randomly divided into control group(Ctrl group) and alcohol group(EtOH group). The EtOH group was fed with Lieber-DeCarli liquid feed for 11 days and then given a single alcohol gavage to establish a chronic and acute alcoholic liver injury model. The Ctrl group was fed with liquid control diet and given isocaloric dextrin by gavage. Serum, liver, intestinal tissue and fecal samples were collected 9 h after gavage. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and triglyceride(TG) in serum and hepatic tumor necrosis factor(TNF)-α, interleukin(IL)-6, IL-8, IL-1β mRNAs were measured. The pathological changes of liver and intestine were observed by HE staining. Illumina highthroughput sequencing was used to detect α diversity, β diversity and species composition at phylum and genus levels.Results Compared with the Ctrl group, alcohol exposure significantly increased serum ALT and AST in mice(P<0.01), and obviously increased liver IL-6, IL-8 and IL-1β mRNA(P<0.01). HE staining showed that there were scattered cell necrosis and inflammatory infiltration in the liver of mice in EtOH group. The intestinal structure was not clear, the gland was atrophy, and there was obvious inflammatory infiltration. The structure of intestinal flora in EtOH group changed,Firmicutesabundance increased at phylum level(P<0.05),Parasutterella abundance decreased at genus level(P<0.05), andClostridiaceaeandClostridium-sensu-strictowere the key genera.Conclusion The composition of intestinal microflora in mice with alcoholic liver injury changes, which may be related to the occurrence of liver disease.

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Objective To explore the role of interleukin(IL)-6 in the process of liver regeneration after acetaminophen(APAP)-induced acute liver injury(ALI) throughin vitroexperiments. Methods In vitroexperiments, AML12 cell lines which originated from normal mice hepatocytes were cultured and treated with different concentrations of APAP drugs and IL-6 neutralizing antibody(IL-6 Ab). CCK-8 method was used to detect cell viability and screen out the appropriate drug effects concentration. The IL-6 concentration was measured by ELISA. Western blot was used to detect the expression levels of liver regeneration-related proteins such as PCNA, CyclinD1, and HGF. The mRNA levels of IL-6, PCNA and CyclinD1 were measured by qRT-PCR. Datas were analyzed by GraphPad Prism 8.0 software. Results Based on the CCK-8 data, the optimal drug concentration of 5 mmol/L APAP was selected for modeling and 0.01 μg/ml IL-6 Ab acting cells. The IL-6 concentration was 0, 1.794, 2.264, 1.658, 1.086 pg/ml at 0, 4, 12, 24, 48 h, respectively, measured by ELISA kit. Western blot results showed that compared with other time points after APAP administration on AML12 cells, the protein levels of CyclinD1 was the highest at 4 h, PCNA and HGF were the highest at 12 h(P<0.05). Compared with the 4 h APAP group, the expression of CyclinD1 and p-STAT3 protein in the APAP+IL-6 Ab group decreased(P<0.05); compared with the 12 h APAP group, the expression of PCNA and HGF in the APAP+IL-6 Ab group decreased(P<0.05). The results of qRT-PCR showed that the mRNA levels of IL-6, PCNA, and CyclinD1 in the 4 h APAP group reached the peak(P<0.05);compared with the 4 h APAP group, the PCNA and CyclinD1 levels in the APAP+IL-6 Ab group were reduced(P<0.05). Conclusion IL-6 Ab inhibits STAT3 phosphorylation by neutralizing IL-6 and subsequently inhibits liver regeneration. It is speculated that IL-6 may play a role in promoting liver regeneration after APAP-induced liver injury.

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Objective To investigate the differential expression of lncRNA SH3BP5-AS1 in hepatocellular carcinoma and the possible mechanism of promoting hepatocarcinogenesis. Methods The expression of SH3BP5-AS1 in hepatocellular carcinoma tissue/paraneoplastic tissue(n=91) was detected by qRT-PCR. TCGA database visualization software UALCAN analyzed SH3BP5-AS1 expression in a large sample of hepatocellular carcinoma tissues(n= 371). The expression of SH3BP5-AS1 was detected by qRT-PCR in normal hepatocytes THLE-2 and hepatoma cell lines Huh 7, BEL-7405, SNU-387, Hep 3 B. After silencing SH3BP5-AS1 expression by siRNA technology, the effects of silencing SH3BP5-AS1 on the proliferation of Hep 3 B cells were detected by CCK-8 assay, cell clone formation assay and Edu assay, and the effects of silencing SH3BP5-AS1 on apoptosis and cycle of Hep 3 B cells were detected by flow-through assay. Based on the expression of SH3BP5-AS1 and TM6 SF2 in hepatocellular carcinoma tissues, the expression correlation was counted and analyzed.Results SH3BP5-AS1 was highly expressed in hepatocellular carcinoma tissues. SH3BP5-AS1 was highly expressed in a large sample of hepatocellular carcinoma tissues from the TCGA database SH3BP5-AS1 was highly expressed in different hepatocellular carcinoma cell lines Huh 7, BEL-7405, SNU-387 and Hep 3 B to a different extent. Silencing of SH3BP5-AS1 by siRNA inhibited the proliferation ability of Hep 3 B cells, promoted apoptosis ability, affected the cell cycle and increased the number of G1 phase cells. SH3BP5-AS1 and TM6 SF2 expression were negatively correlated in hepatocellular carcinoma tissues.Conclusion SH3BP5-AS1 is an important oncogenic non-coding RNA that can provide a theoretical basis for tumour-targeted therapy in patients with hepatocellular carcinoma.

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Objective To investigate the effect and mechanism of pituitary adenylate cyclase activated peptide recombinant 13 peptide(PACAP13) on thymus at cell, organ and individual levels. Methods Thymic epithelial cells(TEC) and bone marrow mesenchymal stem cells(MSC) were culturedin vitro, lymphocyte group, lymphocyte+TEC group, lymphocyte+MSC group and lymphocyte+TEC+MSC group were set up to detect the proliferation of lymphocytes in each group treated by PACAP13 peptide, and contents of cytokines interleukin(IL)-1, IL-2, IL-6 and transforming growth factor(TGF) in culture medium were detected. Dexamethasone(Dex) was added to induce cell apoptosis, and Annexin V-FITC/PI flow cytometry double staining was used to detect the effect of PACAP13 peptide on cell apoptosis. Thymus group, thymus+ TEC group, thymus+ MSC group, thymus+ TEC+ MSC group were set up, and PACAP13 peptide was added, the content of signal joint T cell receptor rearrangement excision cricles(sjTREC) in thymus was detected by real-time quantitative PCR(qRCR). The pristane-induced lupus model mice were randomly divided into model group and PACAP13 group. Model group was injected with 0.9% saline, and PACAP13 group was injected with PACAP13 peptide, thymus and spleen sjTREC levels were detected by qPCR, serum lupus-related autoantibodies were detected by ELISA and thymus-related mRNA expression levels were detected by reverse transcription quantitative PCR(qRT-RCR). Results Compared with the lymphocyte group, the proliferation rate of thymus lymphocytes in the lymphocyte+TEC group, lymphocyte+ MSC group, lymphocyte+ TEC+ MSC group increased, and the apoptosis rate decreased(P<0.05), compared with the control group, the proliferation rate of lymphocytes in the PACAP13 group increased, the rate of apoptosis decreased, and the cytokines IL-1, IL-2, IL-6 and TGF in the cell culture medium increased(P<0.05). Compared with the thymus group, thymus+ TEC group, thymus+ MSC group, thymus+ TEC+ MSC group increased the content of thymus sjTREC, compared with the control group, PACAP13 peptide group thymus sjTREC content increased. Compared with the model group, the content of sjTREC in the thymus and spleen of lupus mice in the PACAP13 group increased(P<0.05), thymus mRNA expression level of AIRE, Fezf2, Foxp3 and TGF-β increased(P<0.01), and CXCL13 mRNA expression level decreased(P<0.01), the autoantibodies of anti-dsDNA antibody, anti-RNP/sm antibody and ANA decreased in serum(P<0.05). Conclusion PACAP13 peptide can promote the proliferation and anti-apoptosis of thymus lymphocytes, enhance the production and output of thymus lymphocytesin vitroandin vivo, and immunomodulatory effects of reducing the level of autoantibodies in lupus mice, and the mechanism is related to the regulation of thymic cytokine secretion and gene transcription level.

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Objective To investigate the ameliorative effects of Mushroom on adipose tissue inflammation and glucolipid metabolism in mice fed a high-fat diet, and to provide a theoretical basis for the mechanisms of Mushroom regulating glucolipid metabolism and inflammatory responses. Methods C57 BL/6 J mice were fed with normal diet(LF) group, high-fat diet(HF)group and high-fat diet + Mushroom(HF+Mushroom) group for 15 weeks.Then, body weight subcutaneous and epididymal white adipose tissue weight were measured. The morphological changes of adipose tissues were compared by HE staining, and the expression of genes related to inflamation, glycolysis and fatty acid oxidation pathways were detected by RT-PCR and Western blot. Results Compared with the LF group, the HF group had increased body weight, increased subcutaneous and epididymal white fat weight and adipocyte size, and upregulated expression of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), monocyte chemoattractant protein-1(MCP-1), CD68, inducible nitric oxide synthase(iNOS), pyruvate kinase(PK), phosphofructokinase(PFK), hypoxia inducible factor-1α(HIF-1α) and peroxisome proliferator activated receptor alpha(PPARα) in adipose tissues, while the expression of carnitine palmitoyl transferase-1 A(CPT-1 A), cytochrome P450 4 a10(CYP4 a10) and medium-chain acyl-coenzyme a dehydrogenase(MCAD) were downregulated(P<0.05). Compared with the HF group, Mushroom supplementation reduced body weight, adipose tissue weight and adipocyte size, and downregulated the expression of pro-inflammatory factors and glycolytic pathway-related factors in adipose tissues, while the expression of fatty acid oxidation pathway-related factors were upregulated(P<0.05). Conclusion Mushroom can ameliorate inflammation and disorders of glycolipid metabolism in adipose tissues of obese mice.

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Objective To explore the effect of eriodictyol(Eri) on the proliferation and apoptosis of triple-negative breast cancer MDA-MB-231 cells and its possible mechanism. Methods MDA-MB-231 cells were treated with different concentrations of Eri. MTT method was used to detect the level of cell proliferation. Plate clone method was used to detect the the level of cell clonality. The apoptosis level was detected by flow cytometry. Western blot was used to detect protein expression levels of Cleaved-caspase-3, Bax, Bcl-2, c-Myc and AR. MDA-MB-231 cells were transfected with si-c-Myc and then combined treatment with 50 μmol/L Eri. MTT method was used to detect the level of cell proliferation. The apoptosis level was detected by flow cytometry. Western blot was used to detect protein expression levels of c-Myc and AR. Results Eri inhibited the proliferation activity of MDA-MB-231 cells in a dose and time-dependent manner. Treatment with different concentrations of Eri reduced cloning ability of MDA-MB-231 cells, promoted cell apoptosis, up-regulated the protein expression levels of Cleaved-caspase-3 and Bax, and down-regulated the protein expression level of Bcl-2, c-Myc and AR. Knockdown of c-Myc expression inhibited MDA-MB-231 cells growth, promoted cell apoptosis, and down-regulated the protein expression level of c-Myc and AR, while combined treatment with 50 μmol/L Eri could not enhance the situation. Conclusion Er inhibits proliferation and induces apoptosis in triple-negative breast cancer MDA-MB-231 cells, which may be achieved by inhibiting c-Myc/AR axis.

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Objective To establish a method for isolation and culture of primary endothelial cells from non-human primate coronary arteries, and to provide a cell model for the study of human coronary endothelial cells. Methods The coronary arteries of macaca mulattas were separated aseptically. The primary endothelial cells were separatedviatissue adhesion after collagenase digestion. CD31 positive cells were detected and sorted by flow cytometry to determine the purity of endothelial cells. After stimulation with prostaglandin E2(PGE2), the cellular viability and proliferation ability of primary coronary endothelial cells from macaca mulattas were evaluated by high-content cell imaging and CCK-8 assay, and the migration ability and tube function of primary coronary endothelial cells from macaca mulattas were measured by Transwell method and Matrigel glue method, respectively. Results The confluence percentage of primary coronary artery cells of macaca mulattas was about 80% after 10-14 daysin vitroculture, and the cellular morphology was irregular polygons and paver shape. The purity of endothelial cells was about 31.7% by flow cytometry. After sorting, the purity of endothelial cells was confirmed by flow cytometry, which was more than 95%. PGE2could significantly up-regulate the proliferation, migration and tube formation abilities of primary coronary endothelial cells of macaca mulattas. Conclusion This study successfully established the isolation and culture method of primary coronary endothelial cells from macaca mulattas, and proved that it could be used as anin vitrocell model to simulate human coronary endothelial cells through functional studies.

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Objective To investigate the effect of human angiopoietin-like protein 4(ANGPTL4) on the invasion and migration of gefitinib-resistant lung adenocarcinoma strains and its mechanism. Methods The expression of ANGPTL4 in PC9/GR cells was interfered by siRNA method, and the interference effect was verified by Western blot and qRT-PCR. The CCK-8 experiment was performed to verify the inhibitory effect of interfering with ANGPTL4 on the proliferation of PC9/GR cells. The scratch experiment and transwell experiment were performed to detect the invasion and metastasis ability of cells; the expression levels of Vimentin, E-cadherin and N-cadherin were detected by Western blot experiment. Results The expression of ANGPTL4 in PC9/GR cells was significantly higher than that in PC9 cells. The CCK-8 experiment proved that the IC50of PC9/GR cells to gefitinib was higher than that of PC9 cells, and the IC50was significantly reduced after interfering with the expression of ANGPTL4 in PC9/GR cells; scratch experiment and Transwell migration experiment showed that after interfering with the expression of ANGPTL4 in PC9/GR cells, the wound healing ability, migration and invasion ability of the cells were significantly reduced. The results of Western blot experiments showed that the expression of N-cadherin and Vimentin decreased and the expression of E-cadherin increased in PC9/GR cells after interference. Conclusion ANGPTL4 may participate in the acquired drug resistance of gefitinib in lung adenocarcinoma, and promote the invasion and metastasis of PC9/GR cells. Interference with ANGPTL4 can inhibit the EMT process of PC9/GR cells.

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Objective To establish colorectal cancer LoVo and SW620 cell lines that stably interfere with the expression of LINC01224, and to explore the effect of down-regulating the expression of LINC01224 on cell apoptosis.Methods The GEPIA2 database was used to analyze the expression of LINC01224 in colorectal cancer tissues; qPCR method was used to detect the expression of LINC01224 in 10 human colorectal cancer cells. Three different LINC01224 siRNAs were respectively transfected into human colorectal cancer LoVo cells, and the LINC01224 shRNA lentiviral vector was constructed with the siRNA sequence with the most obvious inhibitory effect of LINC01224 expression. Recombinant lentiviral particles were packaged in HEK293 T cells and then infected with LoVo and SW620 cells. After selection by puromycin, the monoclonal cells that stably interfere with LINC01224 were obtained by limiting dilution method. MTS method detects cell proliferation ability, and flow cytometry detects cell apoptosis rate.Results The expression of LINC01224 in colorectal cancer tissues was higher than that in normal colorectal tissues, and its expression in 10 types of colorectal cancer cells was also higher than that in normal colorectal epithelial cells HCOEPic. The inhibition rate of siRNA-3 on the expression of LINC01224 in LoVo cells was higher than that of siRNA-1 and siRNA-2. Therefore, siRNA-3 was chosen to design LINC01224 shRNA.Compared with the control group(sh-NC group), the expression level of LINC01224 in the LoVo and SW620 cells of the stable interference LINC01224 group(sh-LINC01224 group) was reduced(P<0.01), and the cell growth rate was slowed down(P<0.01), the rate of apoptosis also increased(P<0.01).Conclusion The shRNA lentiviral interference vector of LINC01224 is successfully constructed, which can stably infect LoVo and SW620 cells, down-regulate the expression of LINC01224 and induce cell apoptosis.

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Objective To investigate the effects of Rab10 and miR-409-3p in diffuse large B-cell lymphoma(DLBCL). Methods qRT-PCR was used to determine miR-409-3p levels following transfection with miR-409-3p mimic, mimic NC, miR-409-3p inhibitor, inhibitor NC and blank control group(control group). Targetscan 7.2 and miRNA target prediction Database(miRDB) software were used to predict the target gene of miR-409-3p. Western blot assay was performed to investigate the expression levels of Rab GTPase10(Rab10). Cell counting Kit-8(CCK-8) and flow cytometry assays were performed to determine the levels of cell viability and apoptosis. Finally, the expression of Rab10 in paraffin-embedded tissues was detected by immunohistochemical(IHC)method, and then the relationship between Rab10 and clinical characteristics in patients with DLBCL was compared. Results miR-409-3p could promote the apoptosis and inhibit proliferation of tumor cells.Overexpression of miR-409-3p increased the Rab10 mRNA and protein levels in DLBCL cells. The expression of Rab10 in DLBCL was higher than that in reactive hyperplasia of lymph nodes, and the level of it was positively correlated with lactate dehydrogenase(LDH), β2-microglobulin(β2-MG) and international prognostic score for lymphoma(IPI) of DLBCL patients. Conclusion miR-409-3p can promote the apoptosis of tumor cells and the expression of Rab10, Rab10 is associated with the adverse prognosis in DLBCL.

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Objective To investigate the expression and therapeutic effects of miR-34a in mice with allergic rhinitis. Methods 10 mice were selected as blank control group, and 30 mice were used to construct mouse allergic rhinitis model with ovalbumin. They were randomly divided into model group, miR-34a inhibitor group and negative control group(NC group) with 10 cases each. 100 μl of physiological saline, 100 μl of negative transfection plasmid, and 100 μl of miR-34a inhibitor carrier mixed solution were injected into the tail vein respectively. The behavioral changes of mice after 1 week of intervention were observed; ELISA was used to detect the level of ovalbumin-specific IgE in mouse serum; qRT-PCR was used to detect interleukin(IL)-6, IL-8, IL-10, and IL-35, the relative expression of matrix metalloproteinase(MMP)-2, MMP-9 and vascular cell adhesion molecule(VCAM)-1 mRNA. The content of CD3+, CD4+, CD8+T cells in peripheral blood was detected by flow cytometry.Results The serum level of miR-34a mRNA in model mice was significantly higher than that in the blank group(P<0.05). Compared with the negative control group, the total behavioral score of mice after miR-34a inhibitor intervention was significantly reduced(P<0.05). The concentration of ovalbumin-specific IgE was significantly reduced(P<0.05), while IL-6, IL-8, IL-10, MMP-2, MMP-9 and VCAM-1 mRNA were significantly reduced(P<0.05), IL-35 increased significantly(P<0.05), and CD3+, CD4+/CD8+increased significantly(P<0.05). Compared with the blank control group, the levels of IL-2 and IFN-γ in the model group decreased(P<0.05); compared with the model group, the levels of IL-2 and IFN-γ in the miR-34a inhibitor group and NC group increased(P<0.05); those indexes of miR-34a inhibitor group and NC group were compared,and there were statistically significant differences(P<0.05); there was no difference between the model group and NC group.Conclusion miR-34a is highly expressed in mice with allergic rhinitis. Inhibition of miR-34a expression can significantly reduce the symptoms and inflammation of model mice, and improve immune function.

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Objective To explore the expression of histone methyltransferase(SETD2) in clear cell renal cell carcinoma(ccRCC) and the influence on prognosis of patients, and to study the possible regulatory mechanism of it. Methods Gene mutation data, transcriptome data and clinical data of patients with ccRCC were downloaded from cancer genome project database(TCGA), and the R software V 4.0.3 was used for statistical analysis of gene expression differences. Kaplan-Meier plotter was used to study the different prognosis of patients with different SETD2 expression and immune infiltration conditions and to reveal the potential mechanism by which SETD2 mutation regulated the progression of ccRCC,meanwhile clinical data were used for verification. Results TIMER2 analysis showed that the expression of SETD2 was significantly down-regulated in ccRCC and related to immune infiltration. The mutation landscape of ccRCC gene showed that the frequency of SETD2 mutation was 12.5%, ranking fourth in somatic mutation frequency. Among patients with ccRCC, the prognosis in the SETD2 high expression group was better than that in the SETD2 low expression group, and the prognosis of patients in the high immune infiltration group was better than that in the low immune infiltration group. Conclusion High expression of SETD2 can increase the infiltration of immune cells and its killing effect on tumor in ccRCC patients, thus playing a positive role in the prognosis of patients.

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Objective To investigate the role and mechanism of mitochondrial permeability transition pore(MPTP) in the damage of hippocampal neurons in SD rats induced by exposure for 6 hours to sevoflurane. Methods The hippocampal neurons of primary cultured SD rats and fetal rats were randomly divided into control group(Con group), cyclosporin A group(CsA group, 0.1,0.5,1 μmol/L), 3% sevoflurane exposure for 6 h Group(Sevo group), cyclosporin A pretreatment+3% sevoflurane 6 h group(Sevo+CsA group, 0.1, 0.5, 1 μmol/L). CCK-8 kit was used to detect cell activity, TUNEL staining was used to detect neuron apoptosis, JC-1 kit was used to detect mitochondrial membrane potential, Calcein AM was used to detect MPTP opening degree, and Western blot was used to detect proteins expression of brain-derived neurotrophic factor(BDNF), postsynaptic compact protein(PSD-95), Snaptophysin, Snapsin-1, and Cyclophilin D(CypD).Results Compared with Con group, the cell viability of Sevo group was significantly reduced(P<0.05), the apoptosis rate of neurons increased(P<0.01),the expression of synapse proteins decreased(P<0.05), the mitochondrial membrane potential decreased(P<0.01), the opening degree of MPTP increased(P<0.01), and the expression of CypD protein increased(P<0.01). Cyclosporin A, a specific inhibitor of MPTP opening, could reduce hippocampal neuron apoptosis and synaptic damage caused by sevoflurane exposure, maintain mitochondrial membrane potential, and inhibit MPTP opening and CypD protein expression.Conclusion Prolonged exposure to sevoflurane can cause damage to hippocampal neurons in SD rats. The mechanism is related to the promotion of CypD-mediated MPTP opening.

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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.

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Objective To investigate the effect of secreted protien acidic and rich in cysteine(SPARC) gene on the proliferation and apoptosis of human keloid fibroblasts and its mechanismin vitro.Methods The lentiviral vector carrying SPARC shRNA(LV2 N-shRNA-SPARC,sh-SPARC group)and empty plasmid(LV2-NC,sh-NC group)were transfected into human keloid fibroblasts(HKF) respectively. Real-time PCR(qRT-PCR) and Western blot were performed to examine the expression of SPARC mRNA and protein in HKF. MTT assay was used to estimate the cell proliferation. The cell cycle and apoptosis of HKF were detected by flow cytometry. The expression of TGF-β1 and TβRⅠ proteins in HKF were detected by Western blot.Results(1) After transfection of SPARC shRNA lentiviral vector, the expression of SPARC mRNA and protein in HKF were significantly down-regulated compared with those in the control groups(P<0.05).(2) MTT assay showed that the proliferation of sh-SPARC groups decreased compared with the control groups(P<0.05).(3) Flow cytometry results showed that the S phase cell ratios of sh-SPARC groups were reduced while the apoptosis rates increased compared with the control groups(P<0.05).(4) Western blot showed that the protein expression levels of TGF-β1 and TβRⅠsignificantly decreased in shSPARC groups.Conclusion SPARC gene silencing inhibits the proliferation blocks cell cycle progression and promotes cell apoptosis in HKF, which may be related to the down-regulation of the TGF-β signaling pathway.

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Objective To establish a sleep deprivation model using zebrafish, and to provide more reliable practical modeling schemes for basic research on insomnia. Methods A total of 160 male 4-month-old wild type AB zebrafish were randomly divided into control group(CK) and sleep deprivation group(SD1-SD7). The control group was placed in a normal 14 h/10 h alternating light/dark environment, and the sleep deprived group was subjected to different days of continuous light with simulated sunlight to achieve the effect of sleep deprivation. After modeling, the changes of movement form, learning and memory ability, biological clock gene expression and brain ultrastructure of zebrafish in each group were compared. Results The results of movement form showed that compared with CK group, the resting time of zebrafish in SD1, SD2 and SD3 groups increased(P<0.05), and the movement time and movement count of zebrafish in SD3 group decreased(P<0.05). The results of learning and memory ability showed that zebrafish in CK group had better learning and memory ability than SD1, SD2 and SD3 groups(P<0.05,0.01), and zebrafish in SD1 group had better learning and memory ability than SD2 and SD3(allP<0.01), and zebrafish in SD2 group had better learning and memory ability than SD3(P<0.01). qRT-PCR results showed that compared with CK group, the mRNA expressions of Per1 a, Per2, Bmal1 and Cry1 b in the brain of zebrafish in SD3 group were significantly different(P<0.05). The results of electron microscopy showed that the neurons of zebrafish brain in SD2 and SD3 groups showed neuronal necrosis. Under the microscope, there were nuclear collapse, boundary cracking and even dissolution, chromatin shrinkage, mitochondrial swelling and even a large number of vacuoles. Conclusion Light exposure for 3 days can change the movement pattern of zebrafish, reduce its learning and memory ability, change the expression of clock genes related to sleep maintenance, and induce the apoptosis of brain neurons, which can be used to establish the model of zebrafish insomnia.

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Objective To investigate the mechanism of down-regulation of the phosphatase and tensin homolog deleted on chromosome ten(PTEN) caused by hepatitis B virus(HBV) infection and the antiviral activity of interferon α(IFN-α). Methods HepG2 cells and HepG2.2.15 cells were cultured under suitable conditions. HepG2 cells were transfected with empty vector(pcDNA3.1) and HBV1.3 plasmid respectively, and the protein expression of PTEN was detected by Western blot; pcDNA3.1 and PTEN over-expression(PTEN-OE) plasmid were transfected into HepG2.2.15 cells respectively. Chemiluminescence was used to analyze the expression of HBV-related antigens in the cell culture supernatant, and real-time quantitative PCR(qRT-PCR) was used to analyze the expression of HBV pregenomic RNA(HBV pgRNA); the synthetic RNA duplex [poly(I∶C)]was used to stimulate PTEN-OE cells, qRT-PCR was used to analyze IFN-α mRNA expression and Western blot was used to analyze the expression of interferon regulatory factor 9(IRF9) protein as well as myxovirus resistance protein 1(MxA) protein in the JAK/STAT signaling pathway.Results In HepG2 cells expressing HBV transiently and HepG2.2.15 cells stably expressing HBV, the expression of PTEN protein both decreased; the expression of HBV-related antigens and HBV pgRNA decreased in PTEN-OE HepG2.2.15 cells compared with the control group. After the treatment by poly(I∶C), the level of IFN-α mRNA was significantly higher than that of the control group, and the expression of IRF9 and MxA ptotein related to the JAK/STAT signaling pathway both increased.Conclusion HBV may play a role in antagonizing the antiviral activity of IFN-α by down-regulating the expression of PTEN.

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Objective To explore the effect of dapagliflozin(DAPA) on the function of rat endothelial progenitor cells(EPCs) culturedin vitroin a high glucose environment. Methods Bone marrow derived EPCs from sprague-dawley(SD) rats were identified by fluorescence staining. EPCs were divided into control group(CG group), high glucose group(HG group), high glucose + DAPA group(GD group) and high glucose + DAPA + LY294002 group(GDL group). MTT assay, flow cytometry, tubule formation assay were used to detect the viability, apoptosis, tubule formation ability of EPCs, respectively. Western blot was used to detect the protein expression of AKT/eNOS signaling pathway. Results Compared with CG group, cell viability, the ability to form tubules, the protein expression of p-AKT and p-eNOS, and the ratio of p-AKT/AKT and p-eNOS/eNOS in HG group significantly decreased(P<0.05), while the apoptosis rate of EPCs significantly increased(P<0.05). Compared with HG group, cell viability, the ability to form tubules, the protein expression of p-AKT and p-eNOS, and the ratio of p-AKT/AKT and p-eNOS/eNOS in GD group significantly increased(P<0.05), while the apoptosis rate of EPCs was significantly reduced(P<0.05). Compared with GD group, cell viability, the ability to form tubules, the protein expression of p-AKT and p-eNOS, and the ratio of p-AKT/AKT and p-eNOS/eNOS in GDL group significantly decreased(P<0.05), while the apoptosis rate of EPCs significantly increased(P<0.05). Conclusion DAPA can protect EPCs from high glucose induced functional damage through AKT/eNOS pathway.

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Objective To explore the effect of cryoablation on apoptosis of lung adenocarcinoma cells. Methods According to the district in the tumor following cryoablation, human lung adenocarcinoma H1299 cells were divided into four groups to construct the freezing model of lung cancer in cell level: group A, untreated cells, as the control group. In group B, the necrotic solution was added to the untreated cell culture medium(the cell suspension was frozen in a liquid nitrogen tank for 5 min, and centrifuged to obtain the supernatant after rewarming at 37 ℃).Group C, sublethal cells(cells were frozen at-80 ℃ for 7 min to mimic sublethal state). In group D, necrotic solution was added into sublethal cell culture medium. Cell apoptosis was observed by flow cytometry analysis 24 h later. The expression of apoptosis related molecules Bax and Bcl-2 was detected by qRT-PCR and Western blot. Lewis lung adenocarcinoma cell line was used to establish subcutaneous transplanted tumor model of C57 BL/6 mice. The successful model mice were randomly divided into two groups: sham operation group and argon-helium cryoablation group, with 4 mice in each group. After argon-helium cryoablation for 14 days, the tumor tissues were taken. The expression of apoptosis-related molecules Bax and Bcl-2 was detected by qRT-PCR and Western blot. Results In the cell freezing model, compared with group A in the control group B,C and D groups could promote cell apoptosis(P<0.05), and group D had the highest apoptosis rate, which was significantly higher than other groups(P<0.05). Bothin vitroandin vivoexperiments showed that cryoablation could increase the expression of Bax mRNA and protein(P<0.05). Bcl-2 mRNA and protein expression decreasedin vivo(P<0.05). There was no significant change in the in cell freezing model. Conclusion Cryoablation can achieve effective ablation through cell apoptosis mechanism, which may be related to the reduction of Bcl-2/Bax.

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Objective To understand the epidemiological and spatial distribution characteristics of mushroom poisoning events in Zunyi, and to provide theoretical basis for the prevention of mushroom poisoning at the grassroots level. Methods Descriptive analysis and spatial distribution of mushroom poisoning events reported by foodborne disease surveillance system in Zunyi were carried out. Results A total of 525 cases of mushroom poisoning events were reported in Zunyi in the past ten years, with a total of 1 758 cases poisoning casesand 29 deaths. The occurrence of mushroom poisoning had seasonal fluctuation, mainly from June to October, accounting for 89.90% of the total. Family was the main place of mushroom poisoning, accounting for 96% of the total. Autochthonous gathering was the main source of mushroom poisoning events. Conclusion Mushroom poisoning is one of the main causes of food poisoning death. With the help of geographic information system(GIS), the distribution map of toadstools is preliminarily explored by combining environmental factors with the distribution of mushroom types.

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Objective To explore the influence of pulmonary lymphatic circulation disorder on the pathogenesis of silicosis. Methods A pulmonary lymphatic circulation disorder model was established by thoracic duct ligation. Twenty-four SPF adult male SD rats were randomly divided into control group, ligation control group, silicosis group and silicosis + ligation thoracic catheter group(hereinafter referred to as silicosis + ligation group) with 6 rats in each group. The control group was left untreated; the rats in the ligation control group were fed with thoracic duct ligation and the rats were fed normally; the rats in the silicosis group were established a silicosis model using dynamic dusting method(the concentration of SiO2dust in the dusting chamber was 2 000 mg/m3, and the rats were given dynamic dusting for 3 hours per day); the rats in the silicosis + ligation group were first surgically ligated to the rat thoracic duct, and then subjected to dynamic dust staining. The conditions for staining were the same as those in the silicosis group. HE staining was used to observe lung tissue pathological changes; Sirius scarlet staining was used to observe collagen Deposition; immunohistochemical observation of pulmonary lymphatic hyperplasia;Western blot was used to detect nuclear factor-κBp65(NF-κBp65), vascular endothelial growth factor receptor-3(VEGFR-3) and α-smooth muscle actin（α-SMA）in rat lung tissue.Results The results of HE and Sirius scarlet staining showed that the lung tissue lesions of the silicosis group and the silicosis + ligation group were more severe compared with the control group. The results of immunohistochemistry showed that there was lymphatic hyperplasia in the lung tissue of the rats in the silicosis group and the silicosis + ligation group. Western blot showed that the protein content of NF-κBp65 and α-SMA in lung tissues were higher in the silicosis + ligation group than that of the silicosis group(2.42±0.05)vs(1.80±0.01),(2.81±0.06)vs(2.57±0.01), respectively. On the contrary, the content of VEGFR-3 in the lung tissue of rats in the silicosis + ligation group was lower than that of the silicosis group(0.87±0.01)vs(0.97±0.03).Conclusion Ligation of the thoracic duct can cause lymphatic circulatory disorders in the lungs of silicosis rats and accelerate the onset of silicosis.

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Objective To explore the cognitive emotion regulation strategies and its influence on quality of life in adult epileptics. Methods A cross-sectional study was conducted on 83 adult epileptics and 53 healthy adults with gender, age and education matching. The Chinese version of cognitive emotion regulation questionnaire(CERQ-C) and the quality of life scale for epileptics(QOLIE-31) were used to evaluate. Results (1) The total score of positive cognitive emotion regulation(PCER) and its sub-items scores in epilepsy group were significantly lower than those in control group(t=-5.587--2.837,P<0.05). The total score of negative cognitive emotion regulation(NCER) and the sub-items scores were significantly higher than those of the control group(t=2.198-4.028,P<0.05).(2) The scores of life quality in epilepsy group were significantly lower than those in control group(t=-7.109--4.226,P<0.001).(3) Pearson correlation analysis showed: the total score of PCER and its sub-items scores were positively correlated with the total score of quality of life(r=0.391-0.581,P<0.001). NCER and its sub-items scores were negatively correlated with the total score of quality of life(r=-0.731--0.540,P<0.001).(4) Multivariate stepwise linear regression analysis showed that seizure worry was negatively correlated with catastrophizing and seizure frequency(t=-3.063--2.574,P<0.05); overall quality of life was positively correlated with catastrophizing(t=-2.214,P<0.05); emotional well being was positively correlated with positive re-attention(t=2.376,P<0.05); emotional well being was negatively correlated with catastrophizing(t=-2.219,P<0.05); fatigue was negatively correlated with blaming others(t=-2.768,P<0.05); cognitive function was positively correlated with positive refocus(t=2.593,P<0.05); medication effects were negatively correlated with blame others(t=-3.348,P<0.05); the total score of quality of life was positively correlated with positive re-attention(t=2.833,P<0.05); the total score of life quality was negatively correlated with catastrophizing(t=-2.729,P<0.05). Conclusion When confronted with negative life events, adult epileptics are more likely to use negative cognitive emotion regulation strategies such as meditation, catastrophizing and blaming others than healthy people, which is closely related to the decline of quality of life of epileptics.

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Objective To investigate the influence of acute myocardial infarction(AMI) combined with hypertension on its circadian rhythm. Methods A total of 1 006 cases of AMI who underwent emergency percutaneous coronary intervention(PCI) surgery were collected continuously, and they were divided into a combined hypertension group and a non-combined hypertension group according to whether it was combined with hypertension. The day was divided into 4 and 12 time periods in units of 6 hours and 2 hours, and the number of cases and differences between the two groups in each time period were compared. Results After propensity score matching(PSM), the two groups had different onsets in the 4 time periods of 0:00—5:59, 6:00—11:59, 12:00—17:59 and 18:00—23:59(P=0.014,0.045,0.035,0.016). After further subdividing the time into 12 time periods in units of 2 hours, the morning peak of the onset time of the hypertensive group was 10:00—11:59(P=0.004), and there was another peak at 2:00—3:59 in the morning(P=0.002). Multivariate Logistic regression indicated that compared with non-combined hypertension, AMI with hypertension had an increased risk of onset in the morning(6:00—11:59)(OR, 1.440; 95%CI, 1.089-1.904;P=0.011). Conclusion Hypertension affects the circadian rhythm of the onset of AMI and the peak time of onset, and it is a risk factor for the onset of AMI in the morning peak period(6:00—11:59).

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Objective To explore the hemorrhagic transformation(HT) and its effect on prognosis in patients with acute ischemic stroke(AIS) after mechanical thrombectomy(MT). Methods A total of 114 patients with AIS received MT were enrolled. The modified Rankin Scale was used to evaluate the clinical outcome at 90 days of onset(0-2 points were good prognosis; 3-6 points were poor prognosis). The patients were divided into HT group(n=25) and non-HT group(n=89) according to their HT conditions. Binomial Logistic regression analysis was performed to determine the vascular risk factors of HT after MT and the effect of HT on prognosis. Results Among 114 patients, there were 25 cases of HT and 89 cases of non HT. The proportion of patients with diabetes in HT group was significantly higher than that in non-HT group. The NIHSS score of HT group at discharge was significantly higher than that in non-HT group. The proportion with good prognosis at 90 days in HT group was significantly lower than that in non-HT group(allP<0.05). Binomial Logistic regression analysis showed that diabetes, high levels of cholesterol and smoking were the major vascular risk factors for HT after thrombectomy(allP<0.05). HT was an important factor affecting the poor prognosis after arterial thrombectomy(P=0.026). Conclusion Diabetes, high levels of cholesterol and smoking are the main risk factors of HT after MT for AIS. HT is an independent risk factor for poor prognosis after MT.

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Objective To preliminarily investigate the mechanism of action of metformin in the treatment of Hashimoto′s thyroiditis. Methods Forty-five healthy SD female rats were randomly divided into control group, model group and metformin-treated group, and the rats were simulated with a model of Hashimoto′s thyroiditis by immuno-injection of high iodine water combined with thyroglobulin and Freund′s adjuvant. After successful modeling, the control group and the model group were perfused with saline, and the metformin group was administered with 300 mg/kg metformin by gavage for 4 weeks, and the rats in each group were executed and sampled. Serum thyroid function indexes(TSH, T3, T4, TGAb, TPOAb) and thyroid tissue inflammatory factors(TNF-α, IL-10, IL-6,IL-12) levels were measured by ELISA; thyroid tissue pathological changes were observed by HE staining; thyroid tissue apoptosis was detected by TUNEL fluorescence staining; thyroid tissue expression of Fas, Fas-L was detected by immunohistochemistry; the expression of Fas, Fas-L, FADD and cleaved caspase-3 in thyroid tissue was determined by Western blot; Fas, Fas-L and FADD mRNA expression in thyroid tissue was detected by qRT-PCR.Results Compared with the control group, serum levels of TSH, TGAb, TPOAb, positive cell ratio, levels of TNF-α, IL-6 and expression of Fas, Fas-L, FADD and cleaved caspase-3 in thyroid tissue were significantly higher in the model group, and serum FT3 and FT4 levels and IL-10 and IL-12 levels in thyroid tissue significantly decreased(P<0.01). Compared with the model group, serum levels of TSH, TGAb, TPOAb, positive cell ratio, TNF-αand IL-6 contents and expression of Fas, Fas-L, FADD and cleaved caspase-3 in thyroid tissue were significantly lower in the metformin group, and serum FT3 and FT4 levels and IL-10 and IL-12 contents in thyroid tissue were significantly higher(P<0.05).Conclusion Metformin inhibits the progression of Hashimoto′s thyroiditis, and its mechanism of action may be related to the inhibition of the Fas/Fas-L-mediated apoptotic pathway.

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Objective To evaluate the differences of serum total light chain(sTLC), urine total light chain(uTLC) and serum free light chain(sFLC) in different stages of chronic kidney disease(CKD) and their correlation with renal function indexes. To investigate the predictive value of light chain indexes in CKD staging. Methods 292 patients with CKD were analyzed retrospectively, and plasma cell diseases, acute kidney injury and tumor diseases were excluded. According to the estimated glomerular filtration rate(eGFR), CKD patients were divided into five groups from CKD 1 stage to CKD 5 stage. The levels of sTLC, uTLC, sFLC and corresponding biochemical indexes of CKD patients were detected, and the differences and correlations among the indexes of each group were compared. The receiver operating curve(ROC curve) was used to analyze the predictive value of each light chain index in CKD stage, with CKD1-2 stage combined as control group and CKD3-5 stage combined as case group. Results There was no significant difference in sTLC κ, sTLC λ, sTLC κ/λ and sFLC κ/λ among CKD1-5 stage(P>0.05). There were significant differences between sFLC κ, sFLC λ and uTLC κ, uTLC λ among CKD1-5 stage(P<0.05), which increased with the increase of CKD staging. The correlation between sFLC κ, sFLC λ and serum creatinine(Scr), blood urea nitrogen(BUN), eGFR were better than uTLC κ, uTLC λ(P<0.001). The sTLC κ, sTLC λ, sTLC κ/λ and sFLC κ/λ had no correlation with renal function indexes(P>0.05). The best critical points of sFLC κ and sFLC λ for predicting CKD3-5 stage were 35.4 mg/L and 52.8 mg/L, and AUC was 0.916(0.883-0.949) and 0.915(0.881-0.949), which were higher than uTLC κ and uTLC λ,AUC was 0.811(0.754-0.869) and 0.787(0.728-0.846), respectively. Conclusion With the increase of CKD staging, the levels of sFLC and uTLC gradually increase. The sFLC and uTLC can effectively predict patients with CKD3 and above, which has an important reference value in stratified management of patients with CKD.

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Objective To investigate the changes of cerebral blood flow(CBF) in patients with type 2 diabetes mellitus(T2 DM) and its correlation with cognitive function and olfactory impairment. Methods Cognitive function assessment and smell identification test were performed on 83 patients with T2 DM and 62 healthy controls(HC). Three-dimensional pseudo-continuous arterial spin labeling(3 D-pcASL) head images were collected from the two groups. CBF values of the cerebral cortex were compared between the patients and HC after the postprocessing. Correlations between the CBF values and cognitive function assessment and between the CBF values and smell identification test scores were analyzed as well. Results Compared to the HC, Chinese smell identification test(CSIT), montreal cognitive assessment(MoCA), digit span test(DST), verbal fluency test(VFT) scores were lower in T2 DM patients(P<0.05).The CBF of the bilateral middle frontal gyrus in T2 DM patients was higher than that in HC group(P<0.001). The CBF of the bilateral gyrus rectus and olfactory cortex in T2 DM patients was lower than that in HC group(P<0.001). Conclusion The cognitive and olfactory function of patients with T2 DM decreased. Patients with T2 DM have abnormal perfusion in the bilateral middle frontal gyrus, gyrus rectus and olfactory cortex, revealing that CBF changes in these brain regions may be one of the causes for cognitive impairment and olfactory dysfunction in T2 DM.

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Objective To compare the effect of cervical tissue expander implantation in the reconstruction of scar deformity after burn with open and endoscopic assistance. Methods 30 patients with faciocervical scar deformity were randomly divided into endoscopic group(n=40) and control group(n=40). The operation time, intraoperative blood loss, postoperative drainage, complication rate, hospital stay and complete expansion time were compared between the two groups. Results Compared with the control group, the average operation time of the endoscopic group was reduced(t=8.09,P<0.05); the intraoperative blood loss and postoperative drainage of the endoscopic group were less than those of the control group, the difference was statistically significant(t=9.97, 5.87,P<0.05); the postoperative hospital stay was reduced, and the difference was statistically significant(t=0.03,P<0.05). Compared with the control group, the incidence of major and minor complications in the endoscopic group was lower than that in the control group(χ2=4.59, 5.17,P<0.05), and the time of complete expansion of water sac in the endoscopic group was reduced(t=3.29,P<0.05). Conclusion The placement of faciocervical tissue expander under endoscope can reduce the operation time, the incidence of postoperative complications, hospital stay and the time to complete expansion. It is worthy of promotion.