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Table of Content

    05 January 2021, Volume 41 Issue 1
    Original Articles
    Effects of SHOX2 over-expression on BMP9-induced osteogenic differentiation of mouse mesenchymal stem cell line C3H10T1/2
    ZHANG Ping, YUAN Xiao-hui, HUANG Hua-kun, YANG Chun-mei, ZHANG Lu-lu, LUO Xiao-ji, LUO Jin-yong
    2021, 41(1):  1-7. 
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    Objective To investigate the effects of short stature homeobox 2(SHOX2) on bone morphogenetic protein 9(BMP9)-induced osteogenic differentiation of mouse mesenchymal stem cell line C3H10T1/2. Methods After construction of SHOX2 recombinant adenoviruse (Ad-SHOX2), normal control(NC) group, Ad-SHOX2 group, BMP9 group, as well as BMP9+Ad-SHOX2 group were set up. Early osteogenic ability was evaluated with alkaline phosphatase(ALP); Late osteogenic ability was detected by Ca2+ deposition; Cell proliferation was observed using microscope; Meanwhile, Western blot was employed to confirm protein expression of proliferating cell nuclear antigen (PCNA), runt-related transcription factor 2(RUNX2), T-ERK1/2 and p-ERK1/2. Results Ad-SHOX2 was constructed successfully. Ad-SHOX2 and BMP9 groups produced higher ALP activity, more Ca2+ accumula-tion, and higher RUNX2 expression in comparison with NC group (P<0.05). Compared with Ad-SHOX2 and BMP9 groups, ALP activity in BMP9+Ad-SHOX2 group was enhanced (P<0.05). Ad-SHOX2(low infection)+BMP9 group dampened Ca2+ accumulation remarkably. And that with high infection stimulated Ca2+ accumulation compared with BMP9 group, but changeless in contrast with Ad-SHOX2 group. Meanwhile, RUNX2 expression in BMP9+Ad-SHOX2 group was higher compared with BMP9 group's, but no change was found in Ad-SHOX2 group (P<0.05). Cell growth density in Ad-SHOX2 and BMP9 groups was greater than NC group's, consistent with protein level of PCNA(P<0.05). And protein expression of PCNA in BMP9+Ad-SHOX2 group was increased compared with BMP9 group's(P<0.05). Additionally, protein expressions of T-ERK1/2 and p-ERK1/2 in both Ad-SHOX2 group and BMP9+Ad-SHOX2 group were up-regulated compared with NC group (P<0.05). Conclusions SHOX2 may promote proliferation and osteogenic differentiation of C3H10T1/2 cells via regulating MAPK/ERK signaling pathway in the absence of exogenous osteoinductive factors. However, its effects on osteogenic differentiation with BMP9 potentially function with antagonistic effect and this is target for further research.
    A case of Kabuki syndrome with precocious puberty and short stature due to novel KDM6A splice-site mutation
    ZHAO Ya-ling, LI Shu-ying, WANG Xi, NIE Min, WU Xue-yan, MAO Jiang-feng
    2021, 41(1):  8-12. 
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    Objective To investigate the pathogenic gene mutations and clinical characteristics of type 2 Kabuki syndrome by analyzing a patient with novel splicing KDM6A gene mutation. Methods Clinical features, laboratory data and radiologic manifestations were collected. The whole exon was sequenced. Results 1)Patient presented with typical facial, skeletal and dermatoglyphic abnormalities, and mild intellectual impairment. 2)The main clinical manifestations were central precocious puberty and short stature. 3)Whole exon sequencing peripheral lymphocytes of the patient and of her parents identified a denovo splicing mutation in KDM6A gene. Conclusions KDM6A mutation may cause Kabuki syndrome, central precocious puberty and dwarfism. This case expands the understanding of clinical manifestations and gene mutation in Kabuki syndrome.
    Effects of adiponectin on the gene promoter activities of fatty acid synthase and hormone sensitive lipase in HepG2 cells
    XU Han-yuan, ZHU Hui-juan, PAN Hui, YANG Hong-bo, WANG Lin-jie, GONG Feng-ying
    2021, 41(1):  13-19. 
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    Objective To investigate the effects of adiponectin on human FAS and HSL gene promoter activities in human hepatocarcinoma cell line HepG2 transfected with plasmids containing human FAS or HSL promoters fused to a luciferase reporter gene. Methods The luciferase reporter gene expression plasmids pGL3-hFAS625-Luc(hFAS 625-Luc) and pGL3-hHSL750-Luc (hHSL 750-Luc) were constructed. HepG2 cells were then transfected with these plasmids by lipofectamine method. The FAS and HSL promoter activities was evaluated by testing the luciferase activity through incubation with 0.5-10.0 μg/mL adiponectin) for 24 h or 5 μg/mL adiponectin for 2-32 h. Results Both hFAS 625-Luc and hHSL 750-Luc plasmids were well expressed in HepG2 cells. 1.0-10.0 μg/mL adiponectin could progressively increase the luciferase expression in HepG2 cells transfected with hFAS 625-Luc and hHSL 750-Luc plasmids with the maximum effect of 1.76 folds (P<0.01) and 1.37 folds of the controls, respectively, at the concentration of 10.0 μg/mL(P<0.01). Moreover, 5 μg/mL adiponectin promoted the expression of luciferase in HepG2 cells transfected with hFAS 625-Luc plasmids at 4 h and 8 h (P<0.01), and inhibited the expression of luciferase at 16 h and 32 h (P<0.01). Adiponectin promoted the expression of luciferase in HepG2 cells transfected with HSL 750-Luc within different durations(2~32 h), and the highest was 1.37 times of the control group at 2 h (P<0.01). Conclusions The luciferase reporter gene plasmids containing human FAS and HSL promoters are successfully constructed. Adiponectin promotes FAS and HSL promoter activities in HepG2 cells with a dose-dependent manner. Adiponectin has a double effect on FAS promoter activity in different times, while it has a promoting effect on HSL promoter activity with different incubation durations.
    Effects of 5-Aza-induced BM-MSCs transplantation on myocardial infarction area, related cytokines and potassium ion channels in rats
    LI Ming-yang, WANG Shu-xia, ZHAO Jing-miao, WANG Qiu-ping, CHEN Ping, JIN Hua, HU Ji-hong
    2021, 41(1):  20-26. 
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    Objective To observe the effects of 5-azacytidine(5-Aza)-induced bone marrow mesenchymal stem cells(BM-MSCs) transplantation on the myocardial infarction area, the expression of related cytokines and potassium channels in rats. Methods Rats were randomly divided into control group, model group, blank control group, BM-MSCs group and 5-Aza-BM-MSCs group. Coronary artery ligation was used to develope a model of myocardial infarction, and cell transplantation was performed using a 4-point injection method. HE staining was used to observe the pathology of infarct myocardium, and the expression of caspase-3, beclin-2, VEGF, PGI2, superoxide dismutase (SOD), IL-3, IL-6 and TNF-α in infarct areas were detected by immunohistochemistry. The Kv1.2 and Kv1.5 proteins expressions of potassium channels were detected by Western blot. Results 1)Compared with the control group, the area of myocardial infarction and the expression level of caspase-3, beclin-2, VEFG, PGI2, IL-3, IL-6 and TNF-α in the model group all significantly increased (P<0.05). However, the expression level of Kv1.2 and Kv1.5 were reduced (P<0.05). 2)Compared with the model group, the area of myocardial infarction and caspase-3, beclin-2, and IL-3, IL-6 and TNF-α expression level significantly reduced both in the BM-MSCs group and the 5-Aza-BM-MSCs group (P<0.05), Kv1.5, VEGF, PGI2, and SOD expression level increased(P<0.05); Kv1.2 in BM-MSCs group decreased, but increased in 5-Aza-BM-MSCs group (P<0.05). 3)Compared with the BM-MSCs group, the area of myocardial infarction and the expression of caspase-3 and IL-6 decreased in the 5-Aza-BM-MSCs group (P<0.05), Kv1.2, VEGF and SOD expression level increased (P<0.05). Conclusions 5-Aza-BM-MSCs more effectively reduces inflammatory response and apoptosis, promotes neovascularization, improves myocardial antioxidant capacity, reduces myocardial infarction area, and promotes the expression of potassium ion channel Kv1.2 protein as compare to single BM-MSCs transplantation.
    Hydroxysafflor yellow A inhibits high glucose-induced renal podocyte injury of mice
    XIE Fei, ZHOU Mei-lan
    2021, 41(1):  27-32. 
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    Objective To investigate the effect of hydroxysafflor yellow A (HYSA) on high glucose (HG)-induced podocyte injury of mice and its mechanism. Methods Cultured podocytes(MCP5) were divided into control group, HG group, HG+HYSA-L group, HG+HYSA-M group and HG+HYSA-H group. Cell viability was detected by CCK-8 method, cell apoptosis was detected by flow cytometry, caspase-3 activity was detected by colorimeter, MDA, SOD and GSH-Px content in cells were detected by kit, the expression of nephrin, fibronectin, α-SMA, VEGF protein and JNK phosphorylation level in cells was detected by Western blot. Results Compared with the control group, the cell survival rate, nephrin protein expression, SOD, GSH-Px content of HG group were significantly reduced, apoptosis rate, caspase-3 activity, MDA content, fibronectin, α-SMA, VEGF protein expression and JNK phosphorylation were all significantly increased (P<0.05); compared with HG group, HYSA reversed the above changes caused by HG with a concentration dependent manner (P<0.05). Conclusions HYSA protects podocytes agains high glucose-induced injury by inhibiting apoptosis, epithelial-mesenchymal transformation and oxidative stress. Its mechanism is potentially related to the inhibition of JNK signal pathway activation and VEGF protein expression.
    Effects of KDM3A silencing on the apoptosis and invasion of human breast cancer cell line MDA-MB-231
    ZHANG Tian-rui, GAO Wen-yi, YAO Juan
    2021, 41(1):  33-37. 
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    Objective To investigate the effect of siRNA mediated KDM3A gene silencing on cell proliferation and apoptosis of breast cancer cell line MDA-MB-231. Methods Constructed a KDM3A gene-specific siRNA lentiviral vector to infect breast cancer MDA-MB-231 cells, and detected the efficiency of breast cancer cells transfected with siRNA-KDM3A by the real-time fluorescent quantitative PCR. CCK-8 method was used to detect cell proliferation, Western blot was used to test cell apoptosis and Transwell method was used to detect cell invasion. Results Compared with the control group,the expression of KDM3A mRNA was down-regulated in KDM3A-sh1, KDM3A-sh2 and KDM3A-sh3 cells, and the most efficient interference RNA was KDM3A-sh2, with the expression of mRNA decreased by 80.1%(P<0.01).After silencing KDM3A, the proliferation of MDA-MB-231 cells decreased, the apoptosis rate increased (P<0.05), and the invasion ability decreased (P<0.01). Conclusions siRNA-mediated KDM3A gene silencing inhibits proliferation and invasion of breast cancer cells and promotes their apoptosis.
    Metabotropic glutamate receptor 5 (mGluR5) regulates white adipocyte browning
    LUO Xie, ZENG Li, YUAN Lei
    2021, 41(1):  38-43. 
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    Objective To investigate whether mGluR5 could regulate white adipocytes browning. Methods Western blot was used to test the expression of mGluR5 in white and brown adipose tissue. Lipid droplets morphology was microscoped after BODIPY staining. Brown adipocytes specific genes and mitochondrial related genes were analyzed by RT-qPCR in mGluR5 antagonists treated 3T3L1 cells. OCR experiments and free glycerol release assay were carried out in 3T3L1 cells after incubation with mGluR5 antagonists. Results mGluR5 expressed in both white and brown adipose tissues. The expression in white fat was higher than that in brown fat(P<0.05). Lipid droplets morphology in adipocytes changed after treatment with mGluR5 antagonists. mGluR5 antagonists promoted the expression of browning genes and mitochondrial related genes, increased mitochondrial respiration rate and promotes lipolysis in 3T3L1 cells. Conclusions Inhibition of mGluR5 may promote white adipocyte browning.
    Proliferation mechanism of acute myeloid leukemia cell line GDM-1 with CSF1R-Y571D mutation
    CAO Qiong, XU Ning-xin, LI Xiao-wen, WU Hao, HAO Jian-qing, QIN Yan-hong, LI Li
    2021, 41(1):  44-49. 
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    Objective To explore the proliferation mechanism of acute myeloid leukemia cell line GDM-1 by the method of protein kinase inhibition. Methods GDM-1 cells were divided into experimental group and control group according to different treatment. Experimental group was treated with 10 different protein kinase inhibitors, the control group was treated with DMSO, and the inhibition of cell proliferation was detected by CCK8 method. The phosphorylation level of key protein kinases in GDM-1 after treatment with linifanib, bosutinib, sorafenib were detected by Western blot.GDM-1 cells were treated with cytokine M-CSF alone or combined with protein kinase inhibitors, cell proliferation was detected by CCK8 method. Results Protein kinase inhibition analysis showed that dasatinib (IC50=0.01 μmol/L), bosutinib(IC50=0.07 μmol/L), linifanib (IC50=0.13 μmol/L) and sorafenib (IC50= 0.12 μmol/L) significantly inhibited the proliferation of GDM-1 cells. Western blot analysis showed that the phosphorylation level of Src and ERK in GDM-1 was decreased (P<0.01) when treated with linifaninb. The proliferation of GDM-1 was 11.3 times as much as that of control group when stimulated with M-CSF. The proliferation of GDM-1 was dramatically inhibited when incubated with the combination of M-CSF and bosutinib (P<0.001), but it was not significantly inhibited when incubated with M-CSF plus HG6-64-1. Conclusions CSF1R-Y571D mutation of GDM-1 cells increases the activity of CSF1R, and promotes cell proliferation by activating Src and MAPK signaling pathways.
    Inflammatory cytokines and apoptosis related proteins participate in the formation of deep vein thrombosis of lower extremity in rats
    FENG Tao, LI Jing, PAN Jin-qiang, ZHENG Dian-yu, CHEN Hui, Duman BAGODAT, GENG Zhong-li
    2021, 41(1):  50-54. 
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    Objective To investigate the expression and regulation mechanism of IL-1β, TNF-α, Bcl-2 and BAX in rats with inferior vena cava thrombosis. Methods The rats were randomly divided into control group, inferior vena cava thrombus model group, mir-5189-3p inhibitor group and miR-5189-3p inhibitor NC group. Twenty-four hours later, the animals were killed by cervical vertebrae dislocation, blood samples were taken and tissues were collected; the level of IL-1β and TNF-α in blood samples were detected by ELISA; The morphology of inferior vena cava thrombus was microscopied with HE staining; The expression of Bcl-2 and BAX protein in inferior vena cava was detected by Western blot. Results The level of IL-1β and TNF-α in the model group was significantly higher than those in the control group (P<0.05); the level of miR-5189-3p inhibitor group was significantly higher than that in the control group (P<0.05); the level of miR-5189-3p inhibitor group was significantly higher than that in the model group (P<0.05); the level of miR-5189-3p inhibitor negative control group was significantly lower than that in the model group (P<0.05). In the model group and miR-5189-3p inhibitor group, red thrombus and mixed thrombus were found; the vessel showed inflammatory cell infiltration, and thrombus in the negative control group of miR-5189-3p inhibitor was obviously organized, with signs of blood flow recanalization. Bcl-2/BAX protein level in the model group was significantly lower than that in the control group (P<0.05); miR-5189-3p inhibitor group was significantly lower than that in the model group (P<0.05); while miR-5189-3p inhibitor in negative control group was significantly higher than that in the model group (P<0.05). Conclusions miR-5189-3p can affect the formation of DVT by regulating the expression of inflammatory cytokines and apoptosis related proteins, which provides a new idea and basis for later research.
    Jagged-1 inhibits mouse embryonic stem cells differentiating into haematopoietic stem or progenitor cells
    CHEN Ri-lin, ZHENG Wei-rong, TAN Lin, LIU Dong-qiang, SHI Hui, CHEN Qi-kang
    2021, 41(1):  55-61. 
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    Objective To investigate the effect of Notch signaling pathways on differentiation of mouse embryonic stem cells(ESC) into haematopoietic stem or progenitor cells(HSC/HPC). Methods 1)incubate mouse embryoid body cells and divide the embryonic stem cells into five groups as EB group,control group,Jagged-1 group, DAPT group and Jagged-1-DAPT group. 2)detect the phenotype of HSC/HPC by flow cytometry. 3)The expression of Notch signaling pathway, mouse embryonic stem cell phenotype and HSC/HPC phenotype were detected by RT-PCR. Results 1)the number of embryonic stem cells in Jagged-1 group was significantly more than that in control group and Jagged-DAPT group(P<0.05);2)the number of HSC/HPC in Jagged-1-DAPT group was significantly more than that in control group and in Jagged-1 group(P<0.05); 3)the expression of Notch1, Notch2 and Notch4 mRNA in Jagged-1 group was significantly higher than that in control group(P<0.05), 4)the expression of Notch1 and Notch4 mRNA in DAPT group and Jagged-1-DAPT group was lower than that in control group(P<0.05). Conclusions Jagged-1 activation of Notch signaling pathway potentially inhibits ESC differentiation into HSC/HPC in mice model.
    Nicotinate-curcumin inhibits LPS-induced inflammatory cytokines secretion and ER stress in HUVECs
    LI Zhao-bing, MA Xiao-feng, JIANG Zhen-tao, HUANG Yun-hui
    2021, 41(1):  62-67. 
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    Objective To investigate the effect of nicotinate-curcumin(NC) on the LPS-induced secretion of cytokines by human umbilical vein endothelial cell (HUVECs) and its mechanism. Methods HUVECs were incubated with different concentrations of NC for 24 h. Western blot was used to detect the phosphorylation of AMPK. After silencing the expression of AMPK with siRNA, the potential outcomes on the phosphorylation of NF-kappa B p65, the expression of adhesion molecules, the secretion of TNF-alpha and MCP-1 in the supernatant of LPS-induced cells, and the expression of endoplasmic reticulum stress markers IRE-1, eIF2α and CHOP were observed. Results The phosphorylation of AMPK was significantly increased and p65 phosphorylation was alleviated after HUVECs treatment with the concentration of 10~20 mmol/L NC for 24 h (P<0.05). After silencing of AMPK, the inhibition effect of NC on NF-κB was significantly reduced(P<0.05). At the same time, the expressions of ICAM-1, VCAM-1 and E-selectin, and the adhesion of THP-1 cells to HUVECs, content of TNF-α and MCP-1 in supernatants were significantly reduced after NC treatment (P<0.05). After siRNA silenced AMPK, the above effects were all inhibited(P<0.05). NC down-regulated the expression of endoplasmic reticulum stress markers IRE-1, eIF2α and CHOP induced by LPS. The inhibitory effect of curcumin on these molecules was significantly reduced by interfering with the expression of AMPK (P<0.05). Conclusions NC inhibits LPS-induced expression, secretion of adhesion molecules and cytokines in HUVECs, and also inhibits endoplasmic reticulum stress. The mechanism is potentially related to the activation of AMPK.
    Effects of main compositions of Astragalus on estradiol synthesizing activing in TGF-β1-induced rat testicular Leydig cell line
    LI Yan-rong, LIU Guang-wei, ZHAO Jing, LI Ya-jun
    2021, 41(1):  68-71. 
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    Objective To investigate the effects of the main compositions of Astragalus on testicular Leydig cells and their synthesis of estradiol(E2), P450arom under TGF-β1 induction. Methods Leydig cells were divided into three groups: control group, model group and Chinese herbal medicine intervention group. Results Compared with the control group, Leydig cells in the model group showed abnormal morphology and less cells(P<0.01).TGF-β1 significantly inhibited the synthesis of E2 and P450arom by Leydig cells (P<0.05). Total Astragalus flavone, total Astragalus saponin and total astragalus polysaccharide all resulted in pathological changes of Leydig cells induced by TGF-β1. Compared with the model group, the number of cells increased significantly (P<0.01) and all of them alleviated the inhibitory effect of TGF-β 1 on the synthesis of E2 in Leydig cells induced by TGF-β1, and the effect of concentration of 20 mg/L total saponin of Astragalus was the most significant. Conclusions Total Astragalus saponin is the most effective component in Astragalus to exert TGF-β1 inhibitory effect on E2 synthesis by Leydig cells.
    Inhibition of inflammatory response by α-lipoic acid in autoimmune thyroiditis mice
    Mayinu YUSUFU, Yibadiguli KUTURUKE, Hanikezi ABUDUAINI
    2021, 41(1):  72-76. 
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    Objective To investigate the inhibitory effect of α-lipoic acid on inflammation in autoimmune thyroiditis (AIT) mice and its effect on lymphocyte subsets. Methods Eighty NOD.H-2h 4 female rats were randomly divided into a control group and the AIT group which was given 50 mg/L sodium iodide orally; Low- and high-dose α-lipoic acid intervention group was injected intraperitoneally with 0.1 and 0.4 g/L α-lipoic acid daily. Twenty animals in each group were continued to be intervened for 8 weeks. After 8 weeks of intervention, HE staining was used to observe lymphocyte infiltration; flow cytometry was used to detect Th17, Treg, CD4+, CD8+ T cells and CD4+/CD8+ levels; and Western blot method was used to determine p38MAPK and NF-κB protein level. Results The incidence of thyroid inflammation in AIT group was as high as 85%. The incidence of thyroiditis, the degree of thyroid inflammation, and the proportion of Th17 cells in the AIT+low/high-dose alpha-lipoic acid group were significantly lower than those in the AIT group (P<0.05), and the proportion of Treg, CD4+, CD8+, and CD4+/CD8+ cells were significant higher than that in the AIT group (P<0.05).The AIT+low/high-dose α-lipoic acid group had significantly lower relative protein expression than those in the AIT group (P<0.05). Conclusions α-lipoic acid can reduce the expression of inflammatory cytokines in autoimmune thyroid mice, improve the proportion of lymphocyte subsets, thereby suppressing the inflammatory response and improving the symptoms of AIT.
    4, 4'-Dimethoxy chalcone improves insulin sensitivity of mice in high-fat diet
    ZHANG Wei-hong, ZHAO Wei, LI Chun-mei, XIE Xiang-hong, YAO Hong, LIU Xiao-jun
    2021, 41(1):  77-81. 
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    Objective To observe the changes of insulin sensitivity of flavonoids 4, 4'-dimethoxy chalcone (DMC) in mice fed on high-fat diet (HFD) and to investigate its effect. Methods Wild-type C57/BL 6 mice fed on HFD were divided into control group and experimental group. DMC and solvent DMSO were injected intraperitoneally at 3 μg/kg every other day. Food intake and body weight of the two groups were recorded weekly, glucose tolerance test (GTT), insulin tolerance test (ITT) and sodium pyruvate tolerance test(PTT) were measured at week 6. The adipose tissues were examined by immunohistochemistry and HE staining. Results Compared with the control mice, the mice treated with DMC drug was no different in body weight and food intake. ITT indicated that DMC administration increased insulin sensitivity (P<0.05), but GTT and PTT showed no significant difference. HE staining microscopy showed that adipose tissue contained less mononuclear cells in mice treated with DMC than control mice. There was weak staining for F4/80 in the adipose tissue of mice treated with DMC, defined as F4/80-positive cells. Conclusions DMC can improve the insulin sensitivity of mice in high-fat diet by decreasing the infiltration of macrophages in adipose tissue.
    Clinical application value of CT portal venography in collateral circulation with portal hypertension
    WU Zhe, TANG Yi, GUO Jun, SUN Hao, JIN Zheng-yu
    2021, 41(1):  82-86. 
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    Objective To evaluate the value of computed tomography portal venography(CTPV) in displaying portal hypertension, and to analyze the imaging findings and related clinical value of different types of portosystemic shunts caused by portal hypertension. Methods A total of 159 patients with portal hypertension were selected from Fushun Central Hospital, including 110 males and 49 females. CT three-dimensional imaging of portal vein was performed at the same time. A total of 297 cases of meaningful shunt branches were selected, and the effects of other shunt branches on esophageal mucosal shunt branches were analyzed. Results All 159 patients had different types of portosystemic communication shunts caused by portal hypertension, including gastric vein group 79.5%(236/297), left renal vein group 7.7%(23/297), umbilical vein recanalization 7.1%(21/297), portal cavernous transformation 2.7%(8/297), intrahepatic shunt 0.7%(2/297) and retroperitoneal group 2.4%(7/297).In most cases, there were two or more types of communicating shunt, the draining vein and shunt branches were clearly dis- played by a three-dimensional imaging of CT portal vein. Conclusions Three-dimensional CT portal vein imaging can show the classification, location, extent and degree of collateral circulation caused by portal hypertension, and effectively track the path of abnormal blood flow, which has important guiding significance for clinical evaluation of the risk of gastrointestinal bleeding and treatment.
    Validation of reference genes for the normalization of the RT-qPCR in peripheral blood mononuclear cells of patients with Takayasu arteritis
    TIAN Yi-xiao, LI Jing
    2021, 41(1):  87-92. 
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    Objective To validate proper reference genes for quantitative real-time polymerase chain reaction (RT-qPCR) used for comparing mRNA expression levels in Takayasu arteritis' (TAK) and healthy controls' (HC) peripheral blood mononuclear cells (PBMC). Methods Total RNA in PBMCs was extracted and used RT-qPCR to determine the profiles of 9 candidate genes, including β-glucuronidase, GAPDH, ACTB, SDHA, HPRT1, RPL13A, B2M, YWHAZ and PKG1. Then compared their transcription stability by geNorm, NormFinder, and BestKeeper. Afterwards, with T-bet, GATA3 and RORC as the targeted genes,explored the influence of reference genes with different stability on mRNA relative abundance. Results The gene combination of B2M-SDHA was selected by geNorm, and HPRT1 was the most stable one in analysis results of NormFinder and BestKeeper, while GAPDH was less stable. Conclusions Genes that have been expressed stably may be upregulated or downregulated when patients with autoimmune diseases received immunosuppressive drugs. When the sample size is small, the more stable internal reference may facilitate the identification of inter-groups difference.
    Clinical Sciences
    Investigation on medication and blood pressure control of hypertension patients in Yuetan community
    LI Xiao-xiao, BIAN Li-li, DONG Yu-ming, LI Juan, LIU Dan-dan
    2021, 41(1):  93-96. 
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    Objective To investigate and analyse the medication and blood pressure control of patients with hypertension in Yuetan community. Methods During June 1, 2018 to January 31, 2019, totally 504 patients with hypertension from the community were investigated, by questionnaire to survey the medication, The blood pressure of patientswere checked by clinic measurement and their history and records were reviewed. Results Systolic blood pressure(SBP)<140 mmHg and diastolic blood pressure(DBP)<90 mmHg, were defined as standard blood pressures in this investigation. The survey showed that the clinic blood pressure compliance rate was 62.3%, the rate of regular medication was 96%, and the proportion of taking one kind of antihypertensive medicine and combining two kinds of antihypertensive medicine was the highest(40.3% and 39.1%, respectively). Among the patients taking three types of antihypertensive medicine, angiotensin-converting enzyme inhibitor(ACEI)/ angiotensin receptor blocker(ARB)(A)+beta blocker(B)+ calcium antagonist(CCB)(C), accounted for the highest proportion (54.8%). The second was the combination of A+C+diuretic(D)(29.0%). Among the two antihypertensive medicine combinations, A+C took the largest proportion(46.2%), followed by B+C(31.5%), while the combination of A+D was lower(9.6%). CCB accounted for the largest proportion (63.5%) in single medicine treatment, followed by ACEI/ARB(26.6%), and diuretic was the lowest (1.0%). Conclusions The proportion of diuretics prescription in hypertension medication in Yuetan community is relatively low, especially in the mono-therapy treatment and in group of patients with two antihypertensive medicines. It is suggested that clinicians should pay more attention to diuretics and its rational use, especially for hypertension patients with high salt intake. In such a situation, low to medium doses thiazide diuretics should be given with priority.
    Clinical characteristics of patients with paroxysmal hypertension caused by panic attack
    FENG Ling, CUI Yun-ying, MA Xiao-sen, WANG Fen, WANG Hui-ping, TONG An-li, LI Yu-xiu
    2021, 41(1):  97-99. 
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    Objective To explore the clinical characteristics of patients with paroxysmal hypertension caused by panic attacks. Methods The clinical data of 24 patients with panic attack treated in Peking Union Medical College Hospital were retrospectively analyzed. Results Nineteen patients presented paroxysmal aggravation of hypertension. Blood pressure of the patients was usually controlled at (136±12/84±9)mmHg with antihypertensive therapy, and rose to (181±13)/(105±15)mmHg during panic attack. The other 5 patients were usually normotensive with average blood pressure (125±10)/(76±6)mmHg, and their blood pressure rose to (175±26)/(92±14)mmHg at panic attack. The surge of blood pressure last from more than ten seconds to six hours. And the attacks were irregular with the frequency ranging from once a few years to several times a day. The attack was triggered by psycho-emotional changes in 54% patients. The proportions of palpitation/panic, dizziness and chest tightness were 92%, 75% and 33% respectively. 24 hours urine catecholamines were normal in all patients. Conclusions Blood pressure of the patients may significantly enhance during panic attacks, mostly accompanying with palpitation/panic, dizziness, chest tightness. The attack may be triggered by psycho-emotional changes. Necessary biochemical tests are needed to exclude other diseases such as pheochromocytoma.
    Mini Reviews
    Research progress of bile acid metabolism in complications of liver cirrhosis
    ZHANG Xue-mei, LI Jun-feng, MAO Xiao-rong
    2021, 41(1):  103-107. 
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    Bile acids are synthesized from cholesterol in hepatocytes and play an important role in the absorption, transport and distribution of fat and fat soluble vitamins. They also act as important signal molecules to regulate metabolism and inflammatory response. The accumulation of bile acids in the body and the activation of bile acid related receptors can aggravate or alleviate the complications of liver cirrhosis such as hepatic encephalopathy, portal hypertension and peritonitis.
    Advances in research on the relationship between intestinal microbiota and frailty
    NIU Xiao-dan, GUO Jing-bo, WANG Hui-lin, CHI Jun-ting, RUAN Hai-hui, TAO Hong-xia, WANG Yan-hong
    2021, 41(1):  108-111. 
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    The quantity of human intestinal microbiota is large and diverse, its changes with age. Intestinal microbiota is closely related to the frailty of the elderly, and can be potential risk factor in the occurrence and development of frailty with different mechanisms. Therefore, the study of the relationship between frailty and intestinal microbiota in elderly patients is of great significance to the regulation of intestinal microbiota in elderly patients with frailty, so that the elderly at risk of frailty can get timely and necessary intervention and help.
    Research progress of thrombus compositions and imaging manifestations in patients with acute ischemic stroke
    LIU Jia-yi, ZHANG Zheng-xiang, YAN Min-li, QI Guan-shu, CHEN Qiang, HOU Qun
    2021, 41(1):  112-115. 
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    With the rapid development of intravascular interventional treatment, the study of thrombus in patients with acute ischemic stroke (AIS) has been intensified in recent vears. The composition of thrombus mainly includes red blood cells, fibrin/platelets, white blood cells, and von Willebrand factor (vWF) and other rare components, which can be manifested as arterial hyperdensity sign (HAS) and susceptibility vessel sign (SVS) on non-contrast CT(NCCT) and magnetic resonance imaging (MRI). Further analysis of thrombus components and imaging manifestations will help to understand the etiology, treatment selection and clinical prognosis of AIS.
    Research advance on telomere shortening and bone marrow failure diseases
    GAO Xiao-yan, LIU Shu-chuan
    2021, 41(1):  116-119. 
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    Telomere shortening is an important pathogenesis of bone marrow failure diseases. As a therapeutic target, telomere may become a new research direction for bone marrow failure diseases. Telomere shortening is related to clinical acquired bone marrow failure diseases, including aplastic anemia, myelodysplastic syndrome and immune mechanism related pancytopenia. The relation between telomere shortening and some inherited bone marrow failure diseases, including dyskeratosis congenita, fanconi anemia is also referred.
    Effect of NLRP3 inflammasome on cerebral ischemia-reperfusion
    GUO Yan-qin, YIN Bo-kai, ZHANG Juan, HE Zhi
    2021, 41(1):  120-124. 
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    NLRP3 inflammasome is a polyprotein complex with complicated structure and unknown regulatory mechanism. Excessive activation of NLRP3 inflammasome is believed to be related to pathophysiological processes with multiple systemic diseases. Cerebral ischemia is the cerebrovascular disease with the highest incidence. Due to the limited application of thrombolytic therapy, it is particularly important to develop neuroprotective drugs after cerebral ischemia. Identification effective drug targets may provide new approaches in the treatment of cerebral ischemia-reperfusion injury(CIRI).
    Summary and prospect of body donation programs in Beijing in the past 20 years
    ZHANG Han-lin, TANG Ke-yun, HU Xin-zhi, ZHANG Qing, WANG Nai-li, ZHANG Di, MA Chao
    2021, 41(1):  125-129. 
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    Cadavers are essential to anatomical dissection courses. The body donation program is the primary source of cadavers used in the courses. In 1999, three body donation programs were founded in Peking Union Medical College, Capital Medical University, and Peking University Health Science Center, which opened a new era for body donation in Beijing. This article summarizes and prospects the body donation programs in Beijing in the past 20 years, and provides a reference for the operation and development of body donation in China.
    Medical Education
    Exploration on the deep integration of “rain classroom” and basic medical teaching
    YAO Qi-ying, ZHANG Dong-mei, ZHAO Hui, WU Xue-fei, ZHU Liang
    2021, 41(1):  130-134. 
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    Objective To explore and practice“rain classroom” in basic medical teaching. Methods Take the undergraduates majoring in Imaging Technology and Biotechnology in the class of 2018 of Dalian Medical University as the research objects Through questionnaire, the teachers and students' feedback on the wisdom teaching of “rain classroom” was investigated, so as to reflect and summarize the auxiliary teaching of “rain classroom”. Results Students believe that “rain classroom” is helpful to their physiology learning and hope to extend this method to use other courses. The teachers impressed on the “rain classroom” exercise answering system,data collection and analysis function which are helpful to understand the learning outcomes of students quantitatively, evaluate the teaching process and adjust the teaching strategy. Conclusions Under the guideline of “Internet +”, “rain classroom” can improve the quality and efficiency of education , encourage students' autonomous learning, promote students' personalized talents training, and thus support the reform of medical education and teaching methodology.
    Kahoot! application research in formative evaluation of human anatomy under SPOC background
    HU Zhuo-yao, XI Jun, HUANG Xiong-feng, HOU Ji-hua, LI Qing, CHEN Qiao
    2021, 41(1):  135-138. 
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    In recent years, the small-scale private online course (SPOC) hybrid teaching model has been widely used in modern education, but its online assessment results often have the problem of obtaining unrealistic data and are not objective and comprehensive. The article makes an in-depth analysis of the above problems, at the same time, the feasibility of Kahoot! application is discussed by taking the human anatomy course as an example, and redesigned the formative evaluation process, aiming to provide a new way for teaching effect evaluation.
    A survey on cognition status, education and practice intention of enhanced recovery after surgery (ERAS) among surgical staffs in Tibet Autonomous Region
    Pingcuoquzhen, LIU Zi-jia, LI Yi, Labaciren, HUANG Yu-guang
    2021, 41(1):  139-143. 
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    Objective To investigate the cognitive status and practice intention of enhanced recovery after surgery (ERAS) among surgical staffs in Tibet Autonomous Region. Methods A questionnaire designed by ourselves was sent to surgical staffs in Tibet Autonomous Region via the network platform. The data were recorded by the system automatically. The full score of the survey was 14 in the cognitive status part. Multinomial logistic regression analysis was used to investigate the related influenc factors of the cognitive status. Results A total of 233 valid questionnaires were collected, with a 97.5% recovery rate. The median age of the participants was 35 years old. The survey covered surgical staffs with different ages and different seniority from all kinds of hospitals and from all the regions in Tibet. The cognition degree of ERAS knowledge was from 3.0% (risk factors of postoperative nausea and vomiting) to 67.4% (preoperative education, nutrition support and medication optimization) among surgical staffs in Tibet Autonomous Region. The median score of the cognitive status part was 4 [4(3,6)], and 51 of them scored 7 or above (21.9%). The results of multinomial logistic regression analysis showed that doctors (regression coefficient=0.349, P<0.05), staffs of anesthesiology department (regression coefficient=0.774, P<0.01) and staffs with senior professional title (regression coefficient=0.634, P<0.05) performed significantly better in the ERAS knowledge cognition. The staffs from general surgical department had significantly better cognitive ability of ERAS than surgical staffs from other departments (P<0.05). 72.9% participants in this study were willing to receive systematic trainings of ERAS, and 74.6% of them were willing to practice ERAS clinically. Conclusions Surgical staffs in Tibet Autonomous Region have poor cognition of perioperative ERAS generally. However, they have strong intention to receive ERAS-related training, as well as enthusiasm of ERAS clinical practice. It is necessary to carry out ERAS education and to explore the management of ERAS in the Tibet Autonomous Region constantly.
    Evaluation of the effect of laparoscopic simulation training on improving operation skills and alleviating anxiety for medical students
    WANG Yi-cheng, WANG Jin-hui, XU Xie-qun, CHEN Ge, LIU Zi-wen
    2021, 41(1):  144-146. 
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    Objective To explore whether the laparoscopic simulation training can improve medical students endoscopy skills and alleviating their anxiety. Methods Students of 8-year program of clinical medicine were recruited without the laparoscopic training in Peking Union Medical College to conduct anxiety questionnaire survey, and perform laparoscopic simulation training. Students who completed 20 hours of training would be assessed and conduct anxiety questionnaire survey again. Results After training, the performance of laparoscopic operation improved (P<0.05), and the degree of anxiety decreased(P<0.05). Conclusions Carrying out laparoscopic simulation training has a good teaching effect.
    Evaluation of simulation training with a stratified and intensive skill training strategy in obstetrics and gynecology
    ZHANG Guo-rui, YU Xin, FAN Qing-bo, LANG Jing-he
    2021, 41(1):  147-150. 
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    Objective This project aimed to evaluate the effect of stratified and intensive training for obstetrics and gynecology skills based on medical simulation technology. Methods The resident trainees and clinical graduates in Peking Union Medical College Hospital received stratified and intensive training of clinical skills in obstetrics and gynecology supported by medical simulation technology. Results After training, the clinical operation skill and occupational capacity of resident trainees and clinical graduates as well as the ability of self-evaluations of trainees were significantly improved. Conclusions Stratified and intensive training of clinical skills supported by medical simulation technology is a feasible and effective training method in the capacity building of residents and clinical graduates in obstetrics and gynecology.
    Application value of “interactive anatomical teaching” combined with “peer education” in the skill training of shoulder joint ultrasound scaning
    FU Shuai, CUI Li-gang, XUE Heng, JIANG Ling
    2021, 41(1):  151-154. 
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    Objective To compare the learning outcomes of “interactive anatomical teaching” combined with “peer education” with traditional teaching in shoulder joint ultrasound training. Methods Two groups of refresher doctors were given shoulder joint ultrasound training, using “interactive anatomical teaching” combined with “peer education” (combined teaching group) and traditional teaching (traditional teaching group)respectively. Before and after the training, selected-response examination, practical operation and film reading ability were evaluated. After the training, a questionnaire survey was conducted to refresher doctors, including comprehensive course evaluation and satisfaction with personal capacity building. Results After the training, there was no statistically significant difference in Objective examination questions between the two groups. The score of combined teaching group was significantly higher than that of traditional teaching group on practical operation and film reading ability (P<0.05). The comprehensive course evaluation and satisfaction with personal ability improvement in combined teaching group was significantly higher than that of traditional teaching group(P<0.05). Conclusions The“interactive anatomical teaching” combined with “peer education” enable the refresher doctors to master the theoretical knowledge, operation skills and diagnosis of shoulder joint ultrasound tecnhology, which is believed to be a better teaching and training method than traditional approaches.