2.Effects of LncRNA-ATB on human peritoneal mesothelial cells phenotypic transition and proliferation induced by high glucose
wen Ming CHE ; Xiang GONG ; jin Xiao ZHANG ; feng Da WEI ; wen Xiao WANG ; min Han WANG ; zhong Li JU
Medical Journal of Chinese People's Liberation Army 2017;42(11):985-991
Objective To explore the effect of long noncoding RNA-ATB (LncRNA-ATB) on phenotypic transition and proliferation of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs used in experiment were divided into three groups:control group,mannitol group and hypertonic glucose group.HPMCs in control group received no treatment,and in hypertonic glucose group and mannitol group were treated with 50mmol/L D-glucose and isotonic mannitol for 72 hours,respectively.Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB,E-cadherin,α-smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),Cyclin D1,cyclin dependent kinase inhibitor 4 (CDK4),protein 27 (p27)and proliferating cell nuclear antigen (PCNA).Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA,CTGF,Cyclin D1,CDK4,p27 and PCNA,and flow cytometry was used to test the cell cycle.Lentivirus artifice was used to up-or down-regulate the expression of LncRNA-ATB in untreated HPMCs.Real-time PCR was employed to detect the mRNA expression of E-cadherin,α-SMA and CTGF,Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA and CTGF,and flow cytometry was used to test the cell cycle.Results It is revealed by Real-time PCR,Western blotting and flow cytometry that the expressions increased of LncRNA-ATB,α-SMA,CTGF,Cyclin D1,CDK4 and PCNA induced by hypertonic glucose,and decreased of E-cadherin and p27 (P<0.05).Up-regulation of LncRNA-ATB promoted HPMCs phenotypic transition and proliferation,while down-regulation alleviated HPMCs phenotypic transition and proliferation.Conclusion Hypertonic glucose may accelerate HPMCs phenotypic transition and proliferation by up-regulating the expression of LncRNA-ATB.
3.Roles of PKCβ/P66Shc oxidative stress signal pathway in mediating hyperoxia-induced ROS production in alveolar epithelial cells.
Zhong-Li CHE ; Wen-Bin DONG ; Qing-Ping LI ; Xiao-Ping LEI ; Lan KANG ; Lin GUO ; Xue-Song ZHAI ; Feng CHEN
Chinese Journal of Contemporary Pediatrics 2015;17(3):275-280
OBJECTIVETo explore the roles of PKCβ/P66Shc oxidative stress signal pathway in mediating hyperoxia-induced reactive oxgen species (ROS) production in alveolar epithelial cells (A549) and the protective effects of PKCβ inhibitor on hyperoxia-induced injuries of alveolar epithelial cells.
METHODSA549 cells were cultured in vitro and randomly divided into three groups: control, hyperoxia and PKCβ inhibitor LY333531 treatment. The hyperoxia group was exposed to a mixture of O2 (950 mL/L) and CO2 (50 mL/L) for 10 minutes and then cultured in a closed environment. The LY333531 group was treated with PKCβ inhibitor LY333531 of 10 µmol/L for 24 hours before hyperoxia induction. Cells were collected 24 hours after culture and the levels of PKCβ, Pin1, P66Shc and P66Shc-Ser36 were detected by Western blot. The intracellular translocation of P66Shc, the production of ROS and cellular mitochondria membrane potential were measured using the confocal microscopy.
RESULTSCompared with the control group, the levels of PKCβ, Pin1, P66Shc and P-P66Shc-Ser36 in A549 cells 24 hours after culture increased significantly in the hyperoxia group. These changes in the hyperoxia group were accompanied with an increased translocation rate of P66Shc from cytoplasm into mitochondria, an increased production of mitochondrial ROS, and a reduced mitochondrial membrane potential. Compared with the hyperoxia group, the levels of Pin1, P66Shc and P66Shc-Ser36 in A549 cells, the translocation rate of P66Shc from cytoplasm into mitochondria and the production of mitochondrial ROS decreased significantly, while the mitochondrial membrane potential increased significantly in the LY333531 treatment group. However, there were significant differences in the above mentioned measurements between the LY333531 treatment and control groups.
CONCLUSIONSHyperoxia can increase the expression of PKCβ in alveolar epithelial cells and production of mitochondrial ROS and decrease mitochondrial membrane potential. PKCβ inhibitor LY333531 can partially disrupt these changes and thus alleviate the hyperoxia-induced alveolar epithelial cell injury.
Cell Hypoxia ; Cells, Cultured ; Epithelial Cells ; metabolism ; Humans ; Indoles ; pharmacology ; Maleimides ; pharmacology ; Oxidative Stress ; Protein Kinase C beta ; physiology ; Pulmonary Alveoli ; cytology ; metabolism ; Reactive Oxygen Species ; metabolism ; Shc Signaling Adaptor Proteins ; physiology ; Signal Transduction ; physiology ; Src Homology 2 Domain-Containing, Transforming Protein 1
4.Inhibitory effects of salviae miltiorrhizae and beta-aescinom natrium on the postburn acute lung injury in rats.
Dian-Cheng TANG ; Jian HOU ; Hong WANG ; Jian-Zhong CHE ; Wen-Song WANG ; Yu-Jie JIA ; Li WANG
Chinese Journal of Burns 2003;19(5):271-274
OBJECTIVETo explore the inhibitory effect and its mechanism of salviae miltiorrhizae and beta-aescinom natrium on the postburn acute lung injury in rats.
METHODSForty-five rats were randomly divided into sham control (C, n = 9), sodium chloride group (S, n = 9), salviae miltiorrhizae group (M, n = 9), beta-aescinom natrium group (A, n = 9), and combination group (MA, n = 9). The rats in M, A and MA groups were subjected to 30% TBSA III degree scald on the back, and all the rats were sacrificed at 24 PBH. The blood and pulmonary tissue samples were harvested from the rats at 24 PBH for the determination of leukocyte adhesiveness/aggregation (LAA) in peripheral blood, myeloperoxidase (MPO), malondialdehyde (MDA) and superoxide dismutase (SOD) contents, and the ratio of wet to dry weights (W/D) of lung tissue.
RESULTSCompared with those in S group, the LAA in blood and the pulmonary tissue contents of MPO, MDA and W/D rate in M and A groups, and especially in MA group, were decreased significantly, but the SOD content in pulmonary tissue increased obviously in M and A groups, especially in MA group. Furthermore, blood LAA was positively correlated with pulmonary tissue MDA content.
CONCLUSIONPostburn intra-pulmonary agglutination and aggregation of PMNs and pulmonary injury by oxygen free radicals (OFRs) and their products could be inhibited by either Salviae Miltiorrhizae or beta-aescinom natrium. In addition, these agents could also increase the tissue content of antioxidant capacity and decrease pulmonary microvascular permeability and lung water content. The results indicated that all the agents used might be effective in prevention and treatment of postburn pulmonary injury, especially when used together.
Acute Lung Injury ; metabolism ; pathology ; prevention & control ; Animals ; Burns ; drug therapy ; metabolism ; pathology ; Escin ; therapeutic use ; Female ; Free Radicals ; metabolism ; Lung ; pathology ; Male ; Nitric Oxide ; metabolism ; Peroxidase ; metabolism ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; Superoxide Dismutase ; metabolism
5.Changes of endothelial cell function and platelet activation in rabbit spinal cord with ischemia-reperfusion injury.
Shou-ping GONG ; Wen-tao WANG ; Da-lin ZHONG ; Jian LV ; Feng WU ; Jin CHE ; Zhi-yuan SENG ; Xi-jing HE
Journal of Southern Medical University 2009;29(8):1638-1640
OBJECTIVETo study the changes of vascular endothelial cell function and platelet activation in rabbit spinal cord following ischemia-reperfusion (I/R) injury and their roles in the spinal cord injury.
METHODSRabbit spinal cord I/R injury models were established using Zivin method, and the changes in plasma NO and GMP140 levels were dynamically monitored after the injury.
RESULTSPlasma NO level increased significantly in the I/R group at the end of the ischemia, and reached the peak level at 2 h of reperfusion as compared to that in sham-operated group (P<0.01). Plasma NO level decreased at 6 h of reperfusion, but still significantly higher than the level in the sham-operated group (P<0.05). Plasma GMP140 underwent no significant changes in the sham-operated group, but significantly increased in the I/R group at the end of the ischemia, followed by gradual declination to the normal level at 2 h of reperfusion.
CONCLUSIONSpinal cord I/R injury causes overexpressions of NO and GMP140, suggesting the involvement of endothelial cell injury and platelet overactivation in the pathological process and repair of spinal cord I/R injury.
Animals ; Endothelial Cells ; metabolism ; Nitric Oxide ; blood ; P-Selectin ; blood ; Platelet Activation ; Rabbits ; Reperfusion Injury ; blood ; pathology ; physiopathology ; Spinal Cord ; pathology ; physiopathology
6.A lightweight multiscale target object detection network for melanoma based on attention mechanism manipulation.
You Wen ZHONG ; Wen Gang CHE ; Sheng Xiang GAO
Journal of Southern Medical University 2022;42(11):1662-1671
OBJECTIVE:
To propose a deep learning target detection model AM- YOLO that integrates coordinate attention and efficient attention mechanism.
METHODS:
Mosaic image enhancement and MixUp mixed-class enhancement were used for image preprocessing. In the target detection model YOLOv5s with One-Stage structure and modified backbone network and neck network, the maximum pooling layer of the spatial pyramid of the backbone network was replaced with a two-dimensional maximum pooling layer, and the coordinate attention mechanism and the efficient channel attention mechanism were integrated into the C3 module and the backbone network of the model, respectively. The improved model was compared with the unmodified YOLOv5s model, YOLOv3 model, YOLOv3-SPP model, and YOLOv3-tiny model for relevant algorithmic indicators in comparative experiments.
RESULTS:
The AM-YOLO model incorporating coordinate attention and efficient channel attention mechanism effectively improved the accuracy of melanoma recognition with also a reduced size of the model weight. This model showed significantly better performance than other models in terms of precision, recall rate and mean average precision, and its mean average precision for benign and malignant melanoma reached 92.8% and 87.1%, respectively.
CONCLUSION
The deep learning-based target object detection algorithm model can be applied in recognition of melanoma targets.
Humans
;
Melanoma
;
Skin Neoplasms
;
Algorithms
;
Software
;
Image Enhancement
7.Effects of Ligustrazine on Airway Inflammation in A Mouse Model of Neutrophilic Asthma.
Xiao-Ming LIU ; Yong-Bin WANG ; Qian WU ; Zhong-Rui BIAN ; Xiao-Wen CHE
Chinese journal of integrative medicine 2018;24(5):353-358
OBJECTIVETo investigate the effects of ligustrazine (LTZ) on airway inflammation in a mouse model of neutrophilic asthma (NA).
METHODSForty healthy C57BL/6 female mice were randomly divided into 4 groups using a random number table, including the normal control, NA, LTZ and dexamethasone (DXM) groups, with 10 rats in each group. The NA mice model was established by the method of ovalbumin combined with lipopolysaccharide sensitization. At 0.5 h before each challenge, LTZ and DXM groups were intraperitoneally injected with LTZ (80 mg/kg) or DXM (0.5 mg/kg) for 14 d, respectively, while the other two groups were given the equal volume of normal saline. After last challenge for 24 h, the aerosol inhalation of methacholine was performed and the airway reactivity was measured. The bronchoalveolar lavage fluid (BALF) was collected. The Wright-Giemsa staining was used for total white blood cells and differential counts. The levels of cytokines interleukin (IL)-17 and IL-10 were detected by enzyme-linked immunosorbent assay. The pathological change of lung tissue was observed by hematoxylin eosin staining.
RESULTSThe airway responsiveness of the NA group was signifificantly higher than the normal control group (P<0.05), while those in the LTZ and DXM groups were signifificantly lower than the NA group (P<0.05). The neutrophil and eosinophil counts in the LTZ and DXM groups were signifificantly lower than the NA group (P<0.05), and those in the LTZ group were signifificantly lower than the DXM group (P<0.05). There were a large number of peribronchiolar and perivascular inflammatory cells in fifiltration in the NA group. The airway inflflammation in the LTZ and DXM groups were signifificantly alleviated than the NA group. The infifiltration in the LTZ group was signifificantly reduced than the DXM group. Compared with the normal control group, the IL-17 level in BALF was signifificantly increased and the IL-10 level in BALF was signifificantly decreased in the NA group (P<0.05). LTZ and DXM treatment signifificantly decreased IL-17 levels and increased IL-10 levels compared with the NA group (P<0.05), and the changes in the above indices were more signifificant in the LTZ group (P<0.05).
CONCLUSIONLTZ could alleviate the airway inflflammation in the NA mice model through increasing the IL-10 level and decreasing the IL-17 level.
Animals ; Asthma ; blood ; complications ; drug therapy ; pathology ; Bronchoalveolar Lavage Fluid ; cytology ; Disease Models, Animal ; Female ; Interleukin-10 ; metabolism ; Interleukin-17 ; metabolism ; Leukocyte Count ; Lung ; drug effects ; pathology ; Mice, Inbred C57BL ; Neutrophils ; drug effects ; pathology ; Pneumonia ; blood ; complications ; drug therapy ; pathology ; Pyrazines ; pharmacology ; therapeutic use ; Respiratory Hypersensitivity ; blood ; complications ; drug therapy ; pathology
8.Diagnostic Value of CD64Index of Neutrophils for Patients with Leukemia Combined with Early Infection.
Jian GUO ; Jun JIA ; Wen-Zhong CHE ; Wen YUAN ; Li-Rui KONG
Journal of Experimental Hematology 2016;24(5):1375-1378
OBJECTIVETo analyze the diagnostic value of CD64index of neutrophils for patients with leukemia combined with early infection.
METHODSFrom June 2014 to June 2015, 100 AL patients admitted in Third People's Hospital of Qinghai province were chosen as AL group, among them 50 AL patients were in infection group and other 50 patients were in non-infection group; at the same time, 200 healthy people were chosen as control group. The sensitivity, specificity and Youden's index of PCT, CRP, Neu% and CD64index in 300 subjects were calculated, and the diagnostic value of different indicators were compared.
RESULTSIn 100 AL patients, the sensitivitys of CD64, Neu%, PCT and CRP were respectively 88.00%, 82.00%, 74.00% and 70.00%; their spesitivitys were respectively 80.00%, 78.00%, 82.00% and 84.00%; their Youden's indexex of were respectively 0.68, 0.60, 0.56 and 0.54.
CONCLUSIONCD64index of neutrophils has diagnostic value for patients with leukemia combined with early infection.