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15 December 2023, Volume 51 Issue 12 Previous Issue    Next Issue
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Cell and Molecular Biology
Experimental Research
Clinical Research
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Cell and Molecular Biology

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Impact of LncRNA TUG1 on high glucose-induced cardiomyocyte apoptosis by regulating the miR-181b-5p/PDCD4 axis LYU Chaoyang, HUANG Ting, XU Zaige, LIU Huishuang, YANG Yingjun, LI Zhenzhen, AO Wen 2023, 51 (12):  1281-1287.  doi: 10.11958/20230523 Abstract ( 216 )   HTML ( 45 )   PDF (1094KB) ( 339 )   Objective To investigate the impact of long non-coding RNA (LncRNA) taurine up-regulated gene 1 (TUG1) on high glucose-induced cardiomyocyte apoptosis by regulating miR-181b-5p/programmed cell death protein 4 (PDCD4) axis. Methods Diabetic cardiomyopathy (DCM) cell model was established in vitro with high glucose (HG,25 mmol/L glucose). AC16 cells were divided into the NG (5.5 mmol/L glucose) group, the HG group, the HG+sh-NC group, the HG+sh-TUG1 group, the HG+miR-NC group, the HG+miR-181b-5p group, the HG+sh-TUG1+anti-miR-NC group, the HG+sh-TUG1+anti-miR-181b-5p group, the HG+miR-181b-5p+pcDNA group and HG+miR-181b-5p+pc-PDCD4 group. The Cell Counting Kit-8 (CCK-8) method was applied to detect cell viability. Lactate dehydrogenase (LDH) assay was applied to detect LDH release. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to detect expression levels of TUG1, miR-181b-5p and PDCD4 mRNA. Flow cytometry was applied to detect apoptosis. Western blot assay was applied to detect levels of B-cell lymphoma 2-associated X (Bax), activated caspase 3 (cleaved caspase 3) and PDCD4 proteins. Caspase-Glo3 assay was applied to assess caspase 3 activity. Dual-luciferase reporter assay was applied to verify the targeting relationship between TUG1 or PDCD4 and miR-181b-5p. Results Compared with the NG group, the cell activity decreased in the HG group, and LDH release, apoptosis rate, Bax, cleaved caspase 3 expression and caspase 3 activity increased (P<0.05), which could be antagonized by TUG1 knockdown or miR-181b-5p overexpression (P<0.05). Inhibition of miR-181b-5p was able to alleviate the impact of TUG1 silencing on cardiomyocyte viability and apoptosis under high glucose treatment (P<0.05). The overexpression of PDCD4 attenuated the promotion effect of miR-181b-5p up-regulation on the viability of cardiomyocytes treated with high glucose and the inhibitory effect on apoptosis. TUG1 was able to increase the expression of PDCD4 through adsorption of miR-181b-5p (P<0.05). Conclusion TUG1 promotes high glucose-induced cardiomyocyte apoptosis by down-regulating miR-181b-5p and up-regulating PDCD4. Figures and Tables | References | Related Articles | Metrics

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Study on the mechanism of genistein inhibiting the progression and metastasis of prostate cancer LIU Wenzhan, CAI Qiliang, WU Baojun, YANG Siwei, YAO Zhili, HOU Zekai, SUN Binxu 2023, 51 (12):  1288-1292.  doi: 10.11958/20230797 Abstract ( 254 )   HTML ( 28 )   PDF (1580KB) ( 332 )   Objective To investigate the effect of genistein on the proliferation, migration and invasion of prostate cancer cells and its molecular mechanism. Methods Prostate cancer LNCaP and CWR22RV1 cells were divided into the control group (conventional culture) and the experimental group (50 μmol/L genistein treatment). The effect of genistein on the proliferation of prostate cancer cells were analyzed by MTT assay. The effect of genistein on the migration and invasion of prostate cancer cells were analyzed by cell scratch assay and Transwell assay. The protein levels of epithelial interstital transformation (EMT) intermediate markers E-Cadherin, N-Cadherin, Vimentin, and tumor stem cell markers CD44 and Oct-4 were detected by Western blot assay. Results MTT assay showed that genistein could inhibit the proliferation of prostate cancer cells. The scratch closure rates of LNCaP and CWR22RV1 cells were significantly reduced in the experimental group compared with those in the control group, and the number of cells passing through the Transwell membrane was significantly reduced (P<0.05). Western blot assay showed that genistein could down-regulate the expression levels of N-Cadherin, Vimentin, CD44 and Oct4 in prostate cancer cells, and up-regulate the expression of E-Cadherin in epithelial cells (P<0.01). Conclusion Genistein reduces the dryness of prostate cancer cells by inhibiting the EMT process, thus reducing the proliferation, migration and invasion of prostate cancer cells. Figures and Tables | References | Related Articles | Metrics

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Impact of shikonin on the malignant biological activity of liver cancer cells by regulating Notch signaling pathway WU Dejian, YANG Qiu, XIE Guidan, PENG Xin 2023, 51 (12):  1293-1299.  doi: 10.11958/20221929 Abstract ( 174 )   HTML ( 22 )   PDF (1452KB) ( 343 )   Objective To investigate the impact of shikonin (SHI) on the malignant biological activity of liver cancer cells by regulating Notch signaling pathway. Methods Western blot assay was used to detect the expression of Notch, Hairy mitosis-related enhancer-1 (Hes1), hairy-related transcription factor-1 (HEY1) protein in liver cancer tissue, paracancerous tissue, hepatoma cells (HepG2 cells, Hep3B cells, HCCLM3 cells, Huh-7 cells and SMMC-7721 cells) and normal liver cells (HL-7702 cells). Huh-7 cells were divided into the control group, the L-SHI group (1 μmol/L SHI), the M-SHI group (2 μmol/L SHI), the H-SHI group (4 μmol/L SHI), the DAPT group (50 μmol/L Notch signal inhibitor DAPT) and the H-SHI+VPA group [4 μmol/L SHI and 8 mmol/L Notch pathway activator Valproic acid (VPA)]. The proliferation of Huh-7 cells was detected by CCK-8 method and plate cloning test. The apoptosis and cell cycle of Huh-7 cells were detected by flow cytometry. Cell scratch test and Transwell invasion test were used to detect migration and invasion of Huh-7 cells. Western blot assay was used to detect the expression of epithelial-mesenchymal transformation (EMT) and apoptosis related proteins. Results The expression levels of Notch, HES1 and HEY1 were obviously increased in liver cancer tissue and cells, and Huh-7 cells showed the most obvious difference, therefore, Huh-7 cells were taken as the research object. Compared with the control group, the protein levels of Notch, HES1, HEY1 and Bcl-2 decreased, and the proportions of S phase and G2 phase cells, OD450 value, number of clones, migration rate, number of invasive cells and levels of N-cadherin and Vimentin decreased significantly in the L-SHI group, the M-SHI group, the H-SHI group and the DAPT group (P<0.05). The proportion of G1/G0 phase cells, apoptosis rate and levels of Bax, cleaved Casase-3, and E-cadherin increased obviously (P<0.05). The effect of SHI was dose-dependent. Compared with the H-SHI group, the above indexes showed the opposite trend in the H-SHI+VPA group. VPA attenuated the effect of SHI on reducing the malignant biological activity of liver cancer cells. Conclusion SHI may inhibit the proliferation, migration and invasion of Huh-7 cells and promote apoptosis of Huh-7 cells by inhibiting Notch signal pathway. Figures and Tables | References | Related Articles | Metrics

Experimental Research

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Impacts of miR-141-3p on pulmonary fibrosis in rats with acute respiratory distress syndrome by regulating Keap1-NRF2/ARE signaling pathway LONG Guangwen, ZHANG Qian, YANG Xiulin, SUN Hongpeng, JI Chunling 2023, 51 (12):  1300-1306.  doi: 10.11958/20230636 Abstract ( 226 )   HTML ( 26 )   PDF (1086KB) ( 327 )   Objective To investigate the effect and mechanism of miR-141-3p on pulmonary fibrosis in rats with acute respiratory distress syndrome (ARDS). Methods Rats were divided into the control group, the model group, the agomir negative control group and the miR-141-3p agomir group according to random number table, with 10 rats in each group. In addition to the control group, the ARDS rat model was established by lipopolysaccharide (LPS) infusion. Rat alveolar type Ⅱ epithelial cells RLE-6TN cells were divided into the NC group, the LPS group, the miR-NC group, the miR-141-3p mimics group, the miR-141-3p mimics+pcDNA group and the miR-141-3p mimics+NRF2 and Kelch-like ring associated protein 1 (Keap1) group. LPS cell model was established in all groups except the NC group. The mRNA expression levels of miR-141-3p and Keap1 in lung tissue and cells were detected by qPCR. Western blot assay was used to analyze lung tissue and cell epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), microtubule associated protein light chain 3B (LC3B), autophagy associated gene Beclin-1, α-smooth muscle actin (α-SMA), type I collagen (Col-Ⅰ), Keap1 and nuclear factors E2 related factor 2 (NRF2) and heme oxygenase 1 (HO-1). HE staining and Masson staining were used to observe pathological changes of lung tissue and to estimate the area of lung tissue injury and pulmonary fibrosis. Hydroxyproline (Hyp) in lung tissue was detected by the kit. Levels of inflammatory factor interleukin-1β, tumor necrosis factor (TNF-α) and oxidative stress index malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by ELISA. Dual luciferase reporting experiment was used to verify the targeting relationship between miR-141-3p and Keap1. Results The expression of miR-141-3p was down-regulated and the expression of Keap1 was up-regulated in lung tissue and cells (P<0.05). Overexpression of miR-141-3p can reduce the degree of pathological damage and fibrosis of lung tissue in rats, Hyp content, and up-regulate expression levels of SOD, E-cadherin, LC3B, Beclin-1, NRF2 and HO-1 in lung tissue and cells, and down-regulate the expression levels of IL-1β, TNF-α, MDA, N-cadherin, α-SMA, Col-I and Keap1 (P<0.05). Overexpression of Keap1 was able to reverse the improvement effect of overexpression of miR-141-3p on alveolar epithelial cell damage in ARDS rats (P<0.05). Double Luciferase reporter gene experiment confirmed that miR-141-3p and Keap1 may have a targeted regulatory relationship. Conclusion Overexpression of miR-141-3p may activate the Keap1-NRF2/ARE signaling pathway, activate autophagy, inhibit inflammatory response, oxidative stress, and EMT progression, and improve pulmonary fibrosis in ARDS rats. Figures and Tables | References | Related Articles | Metrics

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Preliminary study on the effect of intestinal flora changes on glucose metabolism in CG-IUGR rats YUAN Bingshu, LI Lijuan 2023, 51 (12):  1307-1313.  doi: 10.11958/20230026 Abstract ( 201 )   HTML ( 21 )   PDF (943KB) ( 332 )   Objective To analyze the relationship between intestinal flora and glucose metabolism changes in CG-IUGR rats by interventing CG-IUGR rats with the intestinal flora of normal rats. Methods SD rats were divided into three groups: the control group, the CG-IUGR group and the CG-IUGR+intestinal bacteria group (n=8 in each group). The CG-IUGR rat model was established by low-calorie diet. The intestinal flora of rats in the control group were transplanted into CG-IUGR rats in the CG-IUGR+intestinal bacteria group at 3-week-old (once a week, 6 times in total). From birth to 8 weeks, the body weight and body length were measured in three groups of rats, and body mass index (BMI) was calculated. The relevant indexes of glucose metabolism were detected in three groups, including fasting blood glucose (FBG), serum fasting insulin (FINS) and serum insulin (INS) at 15 min after glucose loading and tolerance test of glucose and insulin. The mRNA and protein expression levels of skeletal muscle glycogen synthase (GYS)1 and liver GYS2 were detected by real-time fluorescence quantitative polymerase chain reaction (qPCR) and Western blot assay respectively. The activities of GYS in skeletal muscle and liver were detected. 16S rDNA sequencing was used to detect the diversity and composition of intestinal flora in rats of three groups. Results Compared with the control group, the body length, body weight and BMI of rats in the CG-IUGR group were all increased significantly during the process of growth and development. Its serum FINS level decreased, and serum FBG had no significant change. The blood glucose levels of rats in the CG-IUGR group were significantly up-regulated after glucose and INS loading. The serum INS level of rats in the CG-IUGR group was significantly up-regulated at 15 min after glucose loading. The mRNA and protein expression levels of skeletal muscle GYS1 and liver GYS2 in rats of the CG-IUGR group were decreased, and the activity of GYS was also decreased (P<0.05). These above changes of rats in the CG-IUGR+intestinal bacteria group were all improved (P<0.05). Compared with the control group, ACE and Chao1 indices of intestinal bacteria in rats of the CG-IUGR group were lower (P<0.05). Compared with the CG-IUGR group, the relative abundance of intestinal bifidobacterium in rats of the CG-IUGR+intestinal bacteria group was increased (P<0.05). Conclusion IUGR can attenuate the glucose metabolism fuction of CG-IUGR rats by down-regulating the expression and activity of GYS in skeletal muscle and liver tissue. The abnormal glucose metabolism in CG-IUGR rats is related to changes of intestinal flora. Figures and Tables | References | Related Articles | Metrics

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Effects of Sp5 silencing on Wnt signaling pathway related factors and proliferative ability in mEPMCs BAI Yu, LAN Xuejiao, TANG Jing, WEN Yu, LYU Mingmin, SONG Qinggao 2023, 51 (12):  1314-1320.  doi: 10.11958/20230521 Abstract ( 190 )   HTML ( 189 )   PDF (1448KB) ( 319 )   Objective To investigate the effect of transcription factor specific protein5 (Sp5) silencing on Wnt signaling pathway correlated factors and cell proliferation ability in mouse embryo palatal mesenchymal cells (mEPMCs). Methods mEPMCs of 14.5 d pregnant C57BL/6J mice were isolated and cultured in vitro. Cell source was identified by immunofluorescence staining. Lentivirus transfection technique was used to silence the expression of Sp5 gene in mEPMCs, and the transfection efficiency was verified by Western blot assay. Follow-up experiments were set up with the blank control group, the no-load virus group and the slience group (the Sp5-shRNA group). The protein and mRNA expression levels of β-catenin, GSK-3β, Wnt3a and CyclinD1 were detected by Western blot assay and RT-qPCR after transfection for 72 h in each group. Cell proliferation capacity was detected by CCK-8. The proliferation rate of 5-Ethynyl-2′-deoxyuridine (EdU) positive cells was detected by immunofluorescence assay. Cell cycle was detected by flow cytometry. Results mEPMCs were successfully isolated, and Sp5 expression was silenced. Western blot and RT-qPCR results showed that the protein and mRNA expressions of β-catenin, GSK-3β, Wnt3a and CyclinD1 were significantly higher in the Sp5-shRNA group than those in the blank control group and the no-loaded virus group (P < 0.05). The proliferative ability and the proliferative rate of EdU positive cells were higher in the Sp5-shRNA group than those in the blank control group and the no-loaded virus group (P < 0.05). The proportion of mEPMCs in S phase was higher in the Sp5-shRNA group than that in the blank control group and the no-loaded virus group (P < 0.05). Conclusion Sp5 in silenced mEPMCs can participate in palate development and promote the proliferation of mEPMCs by regulating Wnt signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Study on the mechanism of luteolin reversing multidrug resistance in leukemia K562/ADR cells ZHOU Xinyu, LI Jingmin, ZHANG Ting, JIA Xiuhong 2023, 51 (12):  1321-1325.  doi: 10.11958/20230722 Abstract ( 188 )   HTML ( 24 )   PDF (871KB) ( 305 )   Objective To investigate the mechanism of luteolin′s (Lut) reversal effect on multidrug resistance of chronic myeloid leukemia K562/ADR cells. Methods CCK-8 assay was used to detect drug resistance in K562 and K562/ADR cells 24 hours after treatment with different doses of adriamycin (ADR). CCK-8 assay was used to assess the cytotoxicity and sensitizing effect of Lut on ADR after K562/ADR cells were treated with Lut alone or in combination with ADR for 24 hours. K562/ADR cells in logarithmic growth phase were separated into three group: 0 μmol/L Lut, 2 μmol/L and 4 μmol/L Lut groups. ADR accumulation in cells was measured using flow cytometry. Nuclear factor erythroid-2-related factor 2 (Nrf2), multidrug resistance associated protein 1 (MRP1), P-glycoprotein (P-gp) and glutathione-S-transferase-PI (GST-pi) mRNA and protein expressions were identified using RT-PCR and Western blot assay. Glutathione (GSH) kit was used to detect intracellular GSH content. Results Compared with K562 cells, K562/ADR cell line was significantly resistant to ADR, and the drug resistance was 53.69 times. K562/ADR cell proliferation was decreased to variable degrees by different doses of Lut when compared to the 0 μmol/L Lut group (P<0.05). The proliferation inhibition rates of K562/ADR cells treated with 2 and 4 μmol/L Lut were less than 10%, indicating that the concentration of Lut was non-toxic. Compared with the 0 μmol/L Lut group, the 2 μmol/L Lut group and the 4 μmol/L Lut group showed significantly increased ADR growth inhibition rate on K562/ADR and increased accumulation of ADR in cells, improved the reversal resistance fold, and decreased GSH content in cells. MRP1, P-gp, GST-pi and Nrf2 mRNA and protein expression were reduced in cells (P<0.05). The effect of 4 mol/L Lut was greater than that of 2 mol/L Lut. Conclusion Lut may decrease K562/ADR cell proliferation and reverse ADR medication resistance. The mechanism could be connected to the downregulation of Nrf2, MRP1, P-gp and GST-pi expression, which leads to an increase in ADR accumulation in K562/ADR cells. Figures and Tables | References | Related Articles | Metrics

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The effect and mechanism of exosomes from umbilical cord mesenchymal stem cells on pulmonary inflammation in chronic obstructive pulmonary disease rats NIE Jin, LIU Daishun, ZHANG Jianyong, LIU Chu, ZHOU Liang 2023, 51 (12):  1326-1331.  doi: 10.11958/20230708 Abstract ( 283 )   HTML ( 34 )   PDF (1064KB) ( 308 )   Objective To investigate the effect and mechanism of human umbilical cord mesenchymal stem cell exosomes (Exo) on pulmonary inflammation in chronic obstructive pulmonary disease (COPD) rats. Methods A total of 18 male SD rats were randomly divided into the control group, the COPD group and the COPD + Exo group with 6 rats in each group. COPD rat model was estalished by giving cigarette smoke and lipopolysaccharide (LPS) inhalation in the COPD group and the COPD + Exo group. Rats in the COPD + Exo group were injected with 100 μL of human umbilical mesenchymal stem cell exosome diluent (containing 2 × 107 exosomes) via tail vein. The control group and the COPD group were given equal amounts of phosphate buffered solution. One week after intervention, rats were killed and the peripheral blood, alveolar lavage fluid (BALF) and lung tissue samples were collected. The histopathological changes in lung tissue of each group were observed using Hematoxylin and Eosin (HE) staining. The number of inflammatory cells in peripheral blood was counted by fully automated hematology analyzer. The serum levels of IL-1β, IL-6 and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of IL-1β, IL-6 and TNF-α mRNA in lung tissue were detected by qRT-PCR. Western blot assay was used to detect the protein expression levels of TLR4, p-NF-κB p65 and IκBα in lung tissue. Results In the control group, the structure of pulmonary alveolus was intact and regular, while in the COPD group, the structure of alveolus was disordered, some alveolus expanded irregularly, fused into bullae and even ruptured. The alveolar septa were widened and filled with a large number of inflammatory cells. Compared with the COPD group, the COPD + Exo group had more intact alveolar structure and less infiltration of inflammatory cells in the alveolar septum. The inflammatory cells in peripheral blood decreased. The expression levels of IL-1β, IL-6 and TNF-α in serum and lung tissue were decreased (P < 0.05). The expression of TLR4 and IκBα protein in lung tissue were increased, while the expression of p-NF-κb p65 protein decreased (P < 0.05). Conclusion Human umbilical cord mesenchymal stem cell exosomes can alleviate pulmonary inflammation in COPD rats by modulating TLR4/NF-κB signaling pathway, thereby playing a lung-protective effect. Figures and Tables | References | Related Articles | Metrics

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Study on mechanism of Bupi Yichang pill in alleviating experimental ulcerative colitis by restoring the homeostasis of CD4+T cell subpopulations XIAO Qiuping, ZHAO Chang, LIU Duanyong, LI Shanshan, SHI Min, CHEN Liling, ZHONG Youbao 2023, 51 (12):  1332-1338.  doi: 10.11958/20230776 Abstract ( 154 )   HTML ( 21 )   PDF (1916KB) ( 316 )   Objective To investigate the regulatory effect of Bupi Yichang pill (BPYCP) on CD4+T cell subsets of ulcerative colitis (UC) mice. Methods Forty-eight C57BL/6 mice were randomly divided into 4 groups: the control group (n=10), the model group (DSS group, n=13), the model +BPYCP group (DSS+BPYCP group, n=13) and the model+mesalazine (5-ASA) group (DSS+5-ASA group, n=12). The mouse UC model was induced by 2.5% dextrosan sulfate (DSS) solution. The DSS+BPYCP group and the DSS+5-ASA group were given BPYCP or 5-ASA for 2 weeks, respectively, and fecal viscosity and blood in stool were observed. The colon length was measured. Colonic mass index and unit colonic mass index were calculated. Hematoxylin-eosin (HE) staining was used to observe pathological changes of colon and to score the pathological tissue damage. The level of CD4+T cell subsets in mesenteric lymph nodes was detected by flow cytometry. The expression levels of cytokines interferon-γ (INF-γ), interleukin (IL-4), IL-17A, IL-10 and IL-21 secreted by CD4+T cell subsets in colon tissue were detected by ELISA. Real-time fluorescence quantitative PCR was used to detect colon tissue CD4+T cell subset nuclear transcription factors, mRNA expression levels of T-frame protein 21 (T-bet), GatA-binding protein 3 (GATA-3), retinoa-associated nuclear orphan receptor γt (RORγt), B cell lymphoma-6 (Bcl-6) and Foxp3 in rats. Results Compared with the DSS group, the diarrhea and hematostoecium symptoms of UC mice in the DSS+BPYCP group and the DSS+5-ASA group were significantly improved, body weight and colon length of mice were increased, and colon mass, colon mass index and unit colon mass index were decreased (P<0.05). The mucosal epithelium was more complete than that in the DSS group, and gland arrangement was more regular. The inflammatory cell infiltration was less, and the pathological tissue damage score was significantly decreased (P<0.01). The proportion of Th2 cells in mesenteric lymph nodes was decreased, the proportion of Th17 cells and the level of IL-17A were decreased, and the mRNA levels of T-bet, GATA-3, RORγt and Bcl-6 in colon tissue were decreased (P<0.05). In the DSS+BPYCP group, the proportion of Th1 cells decreased, the proportion of CD4+CD25+Treg cells, CD4+CD25+Foxp3+Treg cells and the level of IL-10 increased, and the proportion of CD4+CXCR5+Tfh cells and the level of IL-21 decreased. The level of Foxp3 mRNA increased (P<0.05). The proportion of Th1 cells and the level of IFN-γ were decreased in the DSS+5-ASA group (P<0.05). Conclusion BPYCP may alleviate UC by remodeling the homeostasis of CD4+T cell subpopulations. Figures and Tables | References | Related Articles | Metrics

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Effect of verbascoside on endothelial dysfunction in atherosclerotic rats by regulating HMGB1/RAGE signal pathway LIU Yanwen, LIU Shuiqing, LIN Shaowei, LIU Xiehong 2023, 51 (12):  1339-1343.  doi: 10.11958/20230325 Abstract ( 204 )   HTML ( 25 )   PDF (874KB) ( 318 )   Objective To investigate the effect of verbascoside (VB) on endothelial dysfunction in atherosclerotic (AS) rats by regulating high-mobility group protein B1 (HMGB1)/receptor for advanced glycation endproducts (RAGE)/nuclear factor κB (NF-κB) signal pathway. Methods The rat model of AS was established by high fat feeding combined with vitamin D3 solution intraperitoneal injection. Rats were divided into the control group (n=10), the model group (n=12), the low (VB-L), medium (VB-M) and high dose (VB-H) VB groups (2, 5 and 10 mg/kg, n=10), and the positive control group (simvastatin, 5 mg/kg, n=10). The serum level of blood lipids was detected by automatic biochemical analyzer. Pathological changes of aorta were observed by HE staining. Serum levels of inflammatory factors and vascular endothelial cytokines were detected by enzyme-linked immunosorbent assay (ELISA). The level of oxidative stress in rats was detected by micro-method kit. The expression of HMGB1/RAGE signal pathway protein in aorta was detected by Western blot assay. Results Compared with the control group, the intima of aorta in the model group was thickened, plaque appeared in blood vessels, accompanied by lipid deposition and inflammatory cell infiltration. Serum levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), C-reactive protein (CRP), matrix metalloproteinase-9 (MMP-9), endothelin-1 (ET-1), visfatin, intercellular adhesion molecule-1 (ICAM-1) and malondialdehyde (MDA), and HMGB1, RAGE and phosphorylation levels of NF-κB in aorta were obviously increased. Serum levels of high-density lipoprotein cholesterol (HDL-C), nitric oxide (NO), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were obviously decreased (P<0.05). Compared with the model group, pathological changes of rats were obviously improved in the VB-L, VB-M and VB-H groups and the simvastatin group. Serum levels of TC, TG, LDL-C, TNF-α, IL-1β, CRP, MMP-9, ET-1, visfatin, ICAM-1, MDA, and HMGB1, RAGE, phosphorylation levels of NF-κB in aorta were obviously decreased, and serum levels of HDL-C, NO, SOD and GSH-Px were obviously increased (P<0.05). Conclusion VB can down-regulate the expression of HMGB1/RAGE/NF-κB signal pathway protein, inhibit inflammation and oxidative stress in AS rats, and improve lipid metabolism and vascular endothelial function. Figures and Tables | References | Related Articles | Metrics

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Improvement effect of myricitrin on bone defect in rats with bacterial osteomyelitis RUAN Jun, LI Xuanying, CHEN Guocai 2023, 51 (12):  1344-1348.  doi: 10.11958/20230188 Abstract ( 212 )   HTML ( 24 )   PDF (948KB) ( 334 )   Objective To explore the improvement effect of myricitrin on bone defect in rats with bacterial osteomyelitis (OM) through vascular endothelial growth factor A (VEGFA)/stromal cell-derived factor (SDF-1)/C-X-C chemokine receptor 4(CXCR4) pathway. Methods Rats were randomly divided into the sham operation group, the OM group, the myricitrin group [50 mg/(kg?d)], PX-478 group [25 mg/(kg?d) VEGFA/SDF-1/CXCR4 signaling pathway inhibitor PX-478], and the myricitrin+PX-478 group [50 mg/(kg?d) myricitrin and 25 mg/(kg?d) PX-478], 18 mice per group. Bilateral proximal tibia bone defects were used to construct bacterial OM rat model. After 12 weeks of continuous treatment, the wound healing degree of rats was observed, and anal temperature of rats was measured. The serum inflammatory factors interleukin (IL)-6, IL-1β, IL-17 and bone metabolism indexes bone alkaline phosphatase (BALP), osteocalcin (BGP) and C-terminal telopeptide of type Ⅰ collagen (CTX-Ⅰ) were measured by enzyma-linked immunosorbent assay (ELISA). The bacterial load of tissue around the lesion was detected. HE staining was used to detect the pathological changes of the tissue around lesions. The expression of CD31 was detected by immunofluorescence staining. Western blot assay was used to detect the expression of VEGFA/SDF-1/CXCR4 signal pathway related proteins. Results Compared with the sham operation group, the number of rats with grade A wound healing, BALP and BGP levels, CD31 positive area, VEGFA, SDF-1 and CXCR4 protein levels were decreased in the OM group (P<0.005 or P<0.05), and the anal temperature, bacterial load, IL-6, IL-1β, IL-17 and CTX-Ⅰ levels were increased (P<0.05). Compared with the OM group, the BALP and BGP levels, CD31 positive area, VEGFA, SDF-1 and CXCR4 protein levels were increased, and the anal temperature, bacterial load, IL-6, IL-1β, IL-17 and CTX-Ⅰ levels were decreased in the myricitrin group (P<0.05). Results of PX-478 group showed the opposite trend. PX-478 reversed the effect of myricitrin on the improvement of bone defects in bacterial OM rats. Conclusion Myricitrin may improve bone defect of OM rats by activating the VEGFA/SDF-1/CXCR4 signal pathway. Figures and Tables | References | Related Articles | Metrics

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Effect of Jiawei Taoren Chengqi Decoction on autophagy in rats with mechanical ventilation-induced lung injury SUN Zhixia, WANG Lihui, SUO Hongliang, CHEN Qian 2023, 51 (12):  1349-1355.  doi: 10.11958/20230374 Abstract ( 235 )   HTML ( 15 )   PDF (1016KB) ( 329 )   Objective To explore the impact of Jiawei Taoren Chengqi Decoction on autophagy in rats with mechanical ventilation-induced lung injury (VILI) through adenylate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. Methods Sixty SPF male rats were randomly grouped into the sham operation group, the model group, the low-dose Jiawei Taoren Chengqi Decoction group (TCM-L group, 2.85 g/kg), the high-dose Jiawei Taoren Chengqi Decoction group (TCM-H group, 8.55 g/kg) and the high-dose Jiawei Taoren Chengqi Decoction + AMPK inhibitor Compound C group (TCM-H+CC group, Jiawei Taoren Chengqi Decoction 8.55 g/kg+ Compound C 250 μg/kg), 12 in each group. Oxygenation index (OI) was measured immediately after intubation and at 1 h, 2 h and 4 h after mechanical ventilation. Bronchoalveolar lavage fluid (BALF) was collected after mechanical ventilation, levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-18 in BALF were detected by enzyme-linked immunosorbent assay (ELISA). Rats were sacrificed, and lung tissue was taken to measure the wet/dry weight (W/D) ratio. HE staining was used to observe pathological changes of lung tissue in each group of rats. Lung injury scores were carried out. Morphology of alveolar epithelial cells was observed by transmission electron microscopy. RT-qPCR was applied to detect mRNA expression levels of AMPK and mTORC1 in rat lung tissue. Western blot assay was applied to detect expression levels of AMPK, p-AMPK, mTORC1, p-mTORC1 and autophagy-related proteins in rat lung tissue. Results Compared with the sham group, the pathological damage of lung tissue was serious, lung W/D, levels of TNF-α, IL-1β and IL-18 in BALF, lung injury score, mTORC1 mRNA expression level, and p-mTORC1 protein expression were increased in the model group (P<0.05). OI values at 2 h and 4 h of mechanical ventilation, AMPK mRNA expression level, p-AMPK, LC3-II/LC3-I and Beclin-1 protein expression in lung tissue were decreased (P<0.05). Compared with the model group, the pathological damage of lung tissue was alleviated in the Chinese medicine-H group, and the trend of changes in related indexes was opposite to the above (P<0.05). Autophagosomes in alveolar epithelial cells were increased. Compound C attenuated the protective effect of Jiawei Taoren Chengqi Decoction on VILI rats (P<0.05). Conclusion Jiawei Taoren Chengqi Decoction may promote autophagy and reduce VILI in rats by activating AMPK/mTOR signaling pathway. Figures and Tables | References | Related Articles | Metrics

Clinical Research

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Analysis of the relationship between IL-36γ level in umbilical cord blood and early infant eczema LU Xujun, WANG Wenge, YANG Yunyue, YANG Yunyan 2023, 51 (12):  1356-1359.  doi: 10.11958/20230643 Abstract ( 192 )   HTML ( 30 )   PDF (792KB) ( 333 )   Objective To investigate the relationship between early infant eczema and cord blood interleukin (IL)-36γ level. Methods Fifty-nine full-term healthy newborns were selected as the study subjects. General information was collected including gender, birth weight, delivery method, gestational age, maternal age, physical fitness during pregnancy (diabetes, infection, hyperthyroidism and other complications), family history of allergies and intake times of seafood during pregnancy ≥3 times. The level of IL-36γ in umbilical cord blood after birth was detected by enzyme-linked immunosorbent assay. The presence and severity of eczema within 42 days were followed up. Multivariate Logistic regression was performed to analyze the influencing factors of early infant eczema, and receiver operating characteristics (ROC) were plotted to evaluate the diagnostic effectiveness. Results Among 59 infants, 40 had eczema, of which 35 were mild, 5 were moderate and 19 were eczema free. The proportion of seafood intake times ≥3 and the level of IL-36γ in umbilical cord blood were higher in the eczema group than those in the no-eczema group (P<0.05). There was no significant difference in IL-36γ level in cord blood between mild and moderate eczema patients. Multivariate Logistic regression analysis showed that the increased level of IL-36γ in umbilical cord blood and ≥3 intake times of seafood during pregnancy were risk factors for early infant eczema (P<0.05). ROC results showed that the AUC (95%CI) of umbilical cord blood IL-36γ was 0.743 (0.611-0.874), sensitivity was 87.6%, specificity was 57.9%, and truncation value was 103.823 ng/L. Conclusion The elevated level of IL-36γ in umbilical cord blood is an independent risk factor for early infant eczema, and early detection is valuable for predicting the occurrence of infantile eczema. Figures and Tables | References | Related Articles | Metrics

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Correlation between serum levels of MMP-13, VASH-1 and prognosis in patients with sepsis complicated with acute kidney injury GUAN Guanghui, PU Qinhua, QIAN Hebu 2023, 51 (12):  1360-1264.  doi: 10.11958/20230928 Abstract ( 256 )   HTML ( 22 )   PDF (779KB) ( 479 )   Objective To investigate the relationship between serum matrix metalloproteinase-13 (MMP-13) and vashohibin-1 (VASH-1) levels and death within 28 days after admission in patients with sepsis complicated with acute kidney injury (AKI). Methods A total of 120 patients with sepsis complicated with AKI (the AKI group) and 117 patients with simple sepsis without complicated AKI (the non-AKI group) were selected, and 136 healthy subjects during the same period were selected as the control group. Basic data of all subjects were collected, and renal function was detected. Serum MMP-13 and VASH-1 levels were detected by enzyme-linked immunosorbent assay. The survival of patients with sepsis complicated with AKI within 28 days after admission was observed. Patients were divided into the survival group (83 cases) and the death group (37 cases). The basic information, renal function, serum MMP-13 and VASH-1 levels were compared between patients in the control group, the non-AKI group, the AKI group and patients in the AKI group with different prognosis. The correlation between serum MMP-13 and VASH-1 levels in patients with sepsis complicated with AKI was analyzed by Pearson method. Logistic regression analysis was used to analyze the factors affecting the death of patients with sepsis complicated with AKI within 28 days after admission. The predictive value of serum MMP-13 and VASH-1 to death within 28 days after admission in patients with sepsis complicated with AKI was analyzed by receiver operating characteristic (ROC) curve. Results Compared with the non-AKI group, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and sequential organ failure assessment (SOFA) score were significantly increased in the AKI group (P<0.05). Compared with the control group and the non-AKI group, serum creatinine (Scr) and blood urea nitrogen (BUN) levels were significantly increased in the AKI group (P<0.05). Compared with the control group, serum levels of MMP-13 and VASH-1 were increased successively in the non-AKI group and the AKI group (P<0.05). Serum MMP-13 and VASH-1 levels were positively correlated with sepsis patients complicated with AKI (r=0.650, P<0.05). Compared with the survival group, APACHE Ⅱ score, SOFA score, serum Scr, BUN, MMP-13 and VASH-1 levels were significantly increased in the death group (P<0.05). APACHE Ⅱ score, serum MMP-13 and VASH-1 levels were independent risk factors for the death in patients with sepsis complicated with AKI within 28 days after admission (P<0.05). The area under curve (AUC) of serum MMP-13, VASH-1 and their combination predict mortality within 28 days after admission in sepsis patients with AKI were 0.810, 0.837 and 0.903, respectively. The AUC predicted by the combination of MMP-13 and VASH-1 was significantly higher than that predicted by serum MMP-13 and VASH-1 alone (P<0.05). Conclusion Serum MMP-13 and VASH-1 levels are increased in patients with sepsis complicated with AKI, which could affect the prognosis of sepsis patients complicated with AKI, and have a high predictive value for death within 28 days after admission. Figures and Tables | References | Related Articles | Metrics

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Clinical features and risk factors of chronic persistent asthma small airway dysfunction PAN Chenhui, WANG Yu, MA Zifeng, WU Dingzhong, ZHANG Shaoyan, QIU Lei, LU Zhenhui 2023, 51 (12):  1365-1368.  doi: 10.11958/20230495 Abstract ( 298 )   HTML ( 17 )   PDF (805KB) ( 415 )   Objective To analyze the clinical characteristics and risk factors of small airway dysfunction (SAD) in patients with asthma. Methods The clinical data of 200 patients with chronic persistent asthma were included, including general data, disease-related condition, pulmonary function test result, compliance assessment and asthma control status. The clinical features of the two groups were compared. Logistic regression was used to analyze risk factors for asthma SAD, and ROC curves were plotted to assess the predictive power of the model. Results Two hundred patients were divided into the SAD group (128 cases) and the non-SAD group (72 cases). The main risk factors of SAD in patients with chronic persistent asthma included smoking history (OR=4.758, 95%CI: 2.043-11.081), overweight (OR=2.952, 95%CI: 1.428-6.105), asthma without clinical remission (OR=6.140, 95%CI: 2.929-12.870), acute asthma attack in recent 1 year (OR=3.406, 95%CI: 1.430-8.117) and allergic rhinitis (OR=2.289, 95%CI: 1.121-4.673). The area under the curve (AUC) of above risk factors were 0.612, 0.610, 0.716, 0.614 and 0.600, respectively. The AUC of the composite prediction model was 0.826 (95%CI: 0.769-0.883), which had good prediction value. Conclusion Smoking, overweight, acute asthma attack in recent one year, non-remission period of asthma and allergic rhinitis are independent risk factors for SAD in chronic persistent asthma. The risk factors of SAD should be identified as early as possible, and individualized monitoring and treatment should be taken. Figures and Tables | References | Related Articles | Metrics

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The correlation between excessive daytime sleepiness and sleep disordered breathing in patients after stroke WANG Wenyi, CHEN Guang 2023, 51 (12):  1369-1373.  doi: 10.11958/20230247 Abstract ( 287 )   HTML ( 15 )   PDF (740KB) ( 576 )   Objective To explore the correlation between excessive daytime sleepiness (EDS) and sleep disordered breathing after stroke. Methods A total of 148 patients with stroke were divided into the EDS group (ESS>7, n=69) and the non-EDS group (ESS≤7, n=79) according to Epworth Sleepiness Scale (ESS). The general data, blood biochemistry and polysomnography (PSG) parameters were compared between the two groups of patients. The correlation between EDS and SDB incidence rate after stroke was analyzed by spearman test. Multivariate Logistic regression analysis was applied for influencing factors of EDS after stroke. Results There were higher proportion of males, hypoventilation index (AHI), oxygen reduction index (ODI), proportion of N1 sleep period and average nighttime diastolic blood pressure in the EDS group than those of the non-EDS group (P<0.05). There was a positive correlation between the incidence rate of EDS and SDB in stroke patients (rs=0.225, P<0.05). Multivariate Logistic regression analysis revealed that gender (OR=2.768, 95%CI: 1.133-6.765), high AHI (OR=1.048, 95%CI: 1.023-1.074) and high average nighttime diastolic blood pressure (OR=1.035, 95%CI: 1.001-1.071) were the independent risk factors for EDS after stroke. Conclusion Male, higher AHI and high average nighttime diastolic blood pressure are independent risk factors for EDS after stroke. Figures and Tables | References | Related Articles | Metrics

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Diagnostic value of ultrasound diagnosis of fetal intrauterine distress in high-risk puerperae in plateau areas ZHAO Yongfeng, GAN Guocai, WANG Xue, ZHAO Xu, MA Shumei, LI Caiqin, ZHANG Cheng 2023, 51 (12):  1374-1377.  doi: 10.11958/20230408 Abstract ( 232 )   HTML ( 17 )   PDF (842KB) ( 335 )   Objective To explore the diagnostic value of color Doppler ultrasound for fetal intrauterine distress (FIUD) in high-risk puerperae in plateau areas. Methods A total of 130 puerperae in plateau areas and 130 puerperae in plain areas were enrolled. According to presence or absence of FIUD in different areas, they were divided into the plateau distress group (47 cases), the plateau normal group (83 cases), the plain distress group (31 cases) and the plain normal group (99 cases). All cases underwent blood flow detection of middle cerebral artery (MCA) and umbilical artery (UA) before delivery, and cerebral -placental ratio (CPR) was calculated. The incidence of FIUD was compared between high-risk puerperae in plateau area and in plain area. Gestational age, birth weight, cesarean section rate and blood spectrum parameters of MCA and UA were compared between the four groups. The predictive value of color Doppler ultrasound parameters for FIUD was analyzed by receiver operating characteristic (ROC) curves. Results The incidence rates of FIUD and severe FIUD were higher in patients of plateau areas than those in plain areas (36.15%, 13.85% vs. 23.85%, 4.62%, P<0.05). Compared with the plateau distress group, gestational age and birth weight were increased in the plateau normal group and the plain distress group (P<0.05). Compared with the plateau distress group, PI, RI, S/D and CPR of MCA were increased, while PI, RI and S/D of UA were decreased in the plateau normal group and the plain distress group (P<0.05). Results of ROC curve analysis showed that overall performance advantage of S/D of UA was the most obvious in the diagnosis of FIUD in high-risk puerperae in plateau areas. The diagnostic sensitivity of RI of MCA was the highest, and the diagnostic specificity of CPR was the highest (P<0.05). Conclusion Color Doppler ultrasound has good diagnostic value for FIUD in high-risk puerperae in plateau areas, which can be applied as an effective clinical screening means for FIUD. Figures and Tables | References | Related Articles | Metrics

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Analysis of clinical characteristics of diabetic ketoacidosis complicated with coronavirus disease 2019 ZHONG Muxian, XIE Xinrong 2023, 51 (12):  1378-1381.  doi: 10.11958/20230432 Abstract ( 315 )   HTML ( 18 )   PDF (733KB) ( 336 )   Objective To explore the clinical features of diabetic ketoacidosis (DKA) complicated with COVID-19. Methods DKA patients were divided into two groups, with 29 cases in the DKA group and 36 cases in the DKA+COVID-19 group. The general data of patients were collected. The serum uric acid, creatinine, white blood cell count, lymphocyte count, hemoglobin, blood glucose at first diagnosis, glycosylated hemoglobin A1c (HbA1c), β-hydroxybutyrate, bicarbonate (HCO3-), arterial blood pH, fasting blood glucose (FBG), 2 h postprandial blood glucose (PBG2 h), fasting C-peptide, 2 h postprandial C-peptide and other laboratory indexes were detected in two groups. The insulin dosage was calculated when blood glucose reached the standard during hospitalization. Results There were no significant differences in blood pressure, gender, body mass index (BMI), uric acid, creatinine, hemoglobin, FBG and PBG2 h between two groups (P>0.05). Compared with the DKA group, patients in the DKA+COVID-19 group were older and had longer course of disease (P<0.05). The white blood cell count, blood glucose at first diagnosis, β-hydroxybutyrate, CK, CK-MB, HbA1c and insulin use were increased, fasting C-peptide, 2 h postprandial C-peptide, PH and lymphocyte count were decreased in the DKA+COVID-19 group (P<0.05). Conclusion Patients with DKA combined with COVID-19 have more severe metabolic disorder and infection at onset, worse immunity, worse islet cell function and higher insulin use. Figures and Tables | References | Related Articles | Metrics

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Establishment and validation of a predictive nomogram model for advanced gastric cancer with lymphovascular invasion GUO Zhenjiang, ZHAO Guangyuan, DU Liqiang, LIU Fangzhen 2023, 51 (12):  1382-1386.  doi: 10.11958/20230513 Abstract ( 310 )   HTML ( 26 )   PDF (853KB) ( 331 )   Objective To explore the preoperative predictors of lymphovascular invasion (LVI) in patients with advanced gastric cancer, and establish the corresponding nomogram prediction model and conduct internal validation. Methods A total of 246 cases of advanced gastric cancer who underwent surgical resection in the Department of Gastrointestinal Surgery of Hengshui People's Hospital from January 2018 to December 2021 were selected. Patients were divided into the LVI positive group and the LVI negative group according to postoperative pathological diagnosis. The age, gender, tumor differentiation, tumor size, tumor site, Borrmann classification, Lauren's classification, cT stage, cN stage and systemic immune-inflammation index (SII) of patients were collected and compared between the two groups. The predictors that were statistically different between the two groups were subjected to multivariate Logistic regression and further developed into a visual prediction model. Bootstrap method was applied for internal validation of the prediction efficiency of the model. Results The differences of tumor size, Borrmann classification, tumor differentiation, Lauren classification, cT staging, cN staging and SII were statistically significant between the two groups ( P<0.05). Multivariate Logistic regression analysis showed that tumor size (OR=2.184, 95%CI:1.224-3.898), Borrmann classification (OR=2.517, 95%CI: 1.294-4.896), cT staging (OR=1.860, 95%CI: 1.045-3.308), cN staging (OR=1.816, 95%CI: 1.004-3.285) and SII (OR=1.001, 95%CI: 1.000-1.002) were independent predictors of LVI in advanced gastric cancer. A preoperative nomogram prediction model for advanced gastric cancer LVI was developed based on results of multivariate analysis. By internal validation, the area under curve (AUC) value of the subject operating characteristic (ROC) curve of the nomogram was 0.735, which was higher than that of tumor size (0.599), Borrmann staging (0.564), cT staging (0.604), cN staging (0.582) and SII (0.615), respectively. The calibration curve showed that the probability of predicted LVI by the nomogram was in a good agreement with the probability of actual LVI occurrence. The Hosmer-Lemeshow test showed good model fit (χ2=4.387, P=0.821). Conclusion The established nomogram prediction model can help to predict the probability of LVI in advanced gastric cancer preoperatively, which can provide a guideline for clinical individualized treatment. Figures and Tables | References | Related Articles | Metrics

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New progress of mechanism of action of miRNA-21 in diabetic kidney disease and Chinese medicine intervention CHEN Yu, HUANG Guodong, QIN Ting, ZHANG Zechao, SHEN Xiaonan, XU Yitan, LIU Shaofang 2023, 51 (12):  1387-1392.  doi: 10.11958/20230945 Abstract ( 284 )   HTML ( 23 )   PDF (755KB) ( 338 )   Diabetic kidney disease is one of the complications of diabetes, which can progress to end-stage renal disease. In recent years, it has been found that miRNAs have become a research hotspot, with miRNA-21 regulating transforming growth factor β1 (TGF-β1)/Smads, phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), Wnt/β-catenin and other signaling pathways to promote the progress of diabetic kidney disease. Studies have showed that traditional Chinese medicine has a regulatory effect on the expression of miRNA-21 and can target miRNA-21 to regulate TGF-β1/Smads, phosphatase and tensin homolog /PI3K/AKT/mammalian target of rapamycin (mTOR), peroxisome proliferator activated receptors and other signal transduction pathways to trigger signal cascade reactions, which intervene in pathological processes such as fibrosis, inflammation, oxidative stress and autophagy. In this article, the role of miRNA-21 in diabetic kidney disease and the intervention of traditional Chinese medicine were summarized, in order to provide some reference for the treatment of diabetic kidney disease and the development of new drugs. References | Related Articles | Metrics