Objective:To investigate the expression of SHIP1 in the patients with acute myeloid leukemia and its effect on the apoptosis of human leukemia cells.Methods:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was detected by Westem blot.U937 cells was transfected with SHIP1 expression vector (pEGFP-SHIP1 group) and empty vector control (pEGFP group) respectively,U937 cells without transfection were used as the control group.Flow cytometry was used to detect the apoptosis of the cells,the expression of SHIP1,Bcl-2,Bax,Akt,p-Akt were detected by western blot.Results:The expression of SHIP1 in the bone marrow of patients with acute myeloid leukemia was significantly lower than that of the normal human bone marrow SHIP 1 (P＜0.01).The SHIP1 and Bax expressions as well as the apoptotic rate ofpEGFP-SHIP1 group were significantly higher than those of the control group(P＜0.01),while the Bcl-2 and p-Akt expressions were significantly lower than those in the control group(P＜0.01).Conclusions:SH-P1 expression was down regulated in the bone marrow of patients with acute myeloid leukemia.SHIP1 could promote the apoptosis of human leukemia cells via Akt signaling pathway.

The canonical transient receptor potential channels (TRPCs) constitute a series of nonselective cation channels with variable degrees of Ca2+ selectivity. TRPCs consist of seven mammalian members, TRPC1, TRPC2, TRPC3, TRPC4, TRPC5, TRPC6, and TRPC7, which are further divided into four subtypes, TRPC1, TRPC2, TRPC4/5, and TRPC3/6/7. These channels take charge of various essential cell functions such as contraction, relaxation, proliferation, and dysfunction. This review, organized into seven main sections, will provide an overview of current knowledge about the underlying pathogenesis of TRPCs in cardio/cerebrovascular diseases, including hypertension, pulmonary arterial hypertension, cardiac hypertrophy, atherosclerosis, arrhythmia, and cerebrovascular ischemia reperfusion injury. Collectively, TRPCs could become a group of drug targets with important physiological functions for the therapy of human cardio/cerebro-vascular diseases.
Arrhythmias, Cardiac ; Atherosclerosis ; Cardiomegaly ; Cardiovascular Diseases ; Cerebrovascular Disorders ; Humans ; Hypertension ; Hypertension, Pulmonary ; Relaxation ; Reperfusion Injury ; Transient Receptor Potential Channels*

Objective To investigate the expression of miR-372 in the plasma of patients with acute my-eloid leukemia(AML)and the possible mechanism to participate in the development of AML. Methods Real-time quantitative PCR was used to detect the level of miR-372 in plasma. Bioinformatics software predicted the pos-sible target genes of miR-372 and dual luciferase reporter assay was performed to validate the prediction. In HL-60 cells,miR-372 was knocked down,and the effects on cell migration and cloning were detected by scratch test and clone formation. Results The level of miR-372 was significantly up-regulated in the plasma of AML patients. ROC analysis showed that miR-372 could distinguish between AML patients and healthy controls. Dual luciferase report-er assay showed that miR-372 could inhibit the activity of PTEN-3'UTR. Inhibition of miR-372 in HL-60 cells can significantly reduce the cell migration rate and clone formation ability. Conclusion In summary,for the first time,we showed novel data that the level of miR-372 was increased in the plasma of AML patients. By targeting the tumor suppressor gene PTEN,miR-372 may become a potential noninvasive biomarker for the screening and di-agnosis of AML.

Objective:To analyze the epidemiological characteristics, high risk factors and pathogenic factors of sepsis-related liver injury patients by collecting epidemiological data and the sequencing results.Methods:A total of 288 sepsis patients been admited to the Emergency Department of the First Affiliated Hospital of Air Force Military Medical University from January 1, 2018 to December 31,2019 were selected and divided into sepsis liver injury group ( n = 44) and sepsis without liver injury group ( n = 244) according to whether acute liver injury occurred or not. The differences ofthe general data, hematological parameters, severity of illness and other indicators at admission between the two groups were compared and analyzed. Logistic regression was used to analyze the risk factors of sepsis-related liver injury. Total of 8 septic patients with liver injury and 4 septic patients without liver injury were selected for RNA-sequencing. Ribonucleic acid (RNA) was extracted from peripheral blood mononuclear cell of patients, detected using RNA-seq, and differential genes were screened and analyzed. Results:Compared with the sepsis without liver injury group, patients in the liver injury group suffered less hypertension (11.4% vs. 30.3%) and relatively more chronic renal insufficiency (40.9% vs. 12.1%); more patients were admitted to the emergency department due to renal disease (43.2% vs. 24.6%), higher sequential organ failure score (SOFA) and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score (SOFA (points) 9.86 ± 3.59 vs. 5.41 ± 3.13, APACHE Ⅱ (points) (16.07 ± 4.41) vs. (14.46 ± 3.77), with prolonged hospital days (d): 8 (4.75, 13.75) vs. 6 (2, 9)]; in the liver injury group, the incidence of infection in respiratory and digestive systems (70.5% vs. 18.0%) andthe chance of infection with Staphylococcus aureus were higher (9.1% vs. 2.0%), and laboratory parameters (procalcitonin (PCT), lactate dehydrogenase (LDH), partial thromboplastin time (APTT), direct bilirubin (DBIL), aspartate aminotransferase (ALT), alanine aminotransferase (AST)) were significantly increased [PCT (μg/L) (23.90 ± 33.22) vs. (10.95 ± 20.18), LDH (U/L) 540.00 (370.50, 1177.00) vs. 168.00 (98.65, 875.18), APTT (s) (41.50 ± 3.13) vs. (36.23 ± 5.27), DBIL (μmol/L) 18.50 (10.10, 58.85) vs. 10.30 (7.60, 16.85), ALT (U/L) 67.00 (41.25, 164.00) vs. 29.00 (18.00, 51.25), AST (U/L), 101.00 (51.25, 174.75) vs. 35.00 (25.00, 65.50)], while platelet (PLT) and albumin (Alb) were significantly lower than those in the sepsis without liver injury group [PLT (× 10 9/L) 62.50 (38.50, 164.25) vs. 90.5 (66.25, 165.5), Alb (g/L) (30.17 ± 7.16) vs. (34.20 ± 6.50)] (all P < 0.05).Logistic regression analysis revealed that Staphylococcus aureus infection, thrombocytopenia, elevated procalcitonin, elevated lactate dehydrogenase, elevated total bilirubin, and elevated glutamyltransferase were associated with sepsis with acute liver injury (odds ratio, OR) with 95% confidence interval (95% CI) of 0.1167 (0.0380~0.7300), 0.9836 (1.0060~1.0290), 0.9986 (1.0000~1.0001), 0.9745 (1.0040~1.0170), 1.0020 (0.9940~1.0000), and 0.9931 (1.0000~1.0001), respectively. A total of 311 significantly differential expressed genes (DEGs) were selected, with 151 up-regulated genes and 160 down-regulated genes compared with the septic non-liver injury group. Further bioinformatics analysis reveled that the top 10 GO sequences are:①platelet α granules,② platelet α granule cavity,③wound healing,④cell migration,⑤multicellular organism process,⑥anatomical structure development,⑦cartilage ossification,⑧tissue development,⑨ keratinization,⑨Multicellular biological development. And KEGG pathway enrichment analysis revealed that human disease-related pathways were dominant, mainly including purine metabolism, AGE-RAGE signaling pathway, p53 signaling pathway, porphyrin and chlorophyll metabolism, nitrogen metabolism, mineral nutrient absorption, protein processing in the endoplasmic reticulum, and FoXo signaling pathway. Conclusions:Staphylococcus aureus infection, thrombocytopenia, elevated procalcitonin, elevated lactate dehydrogenase, elevated total bilirubin, and elevated glutamyltransferase were independent risk factors for sepsis liver injury. Coagulation dysfunction, apoptosis, and metabolic level changes may be important mechanisms of sepsis-associated liver injury, which are related to purine metabolism, porphyrin and chlorophyll metabolism and the expression of genes related to FoXo signaling pathway, Hippo signaling pathway, and p53 signaling pathway.

Adult ; Bone Neoplasms ; diagnosis ; pathology ; Chondrosarcoma ; diagnosis ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Mediastinal Neoplasms ; diagnosis ; pathology ; Neurilemmoma ; diagnosis ; pathology ; Peripheral Nervous System Neoplasms ; diagnosis ; pathology ; Phrenic Nerve ; Ribs ; Tomography, X-Ray Computed

Objective To explore the feasibility of using the optimization parameters modification and optimization processes modification to reduce the total monitor units ( MUs ) in the Eclipse radiotherapy treatment planning system (TPS). Methods Based on the radiotherapy plan of 10 patients with nasopharyngeal carcinoma, a total of 90 plans were designed for 9 groups using different optimization parameters and processes. The total MUs and the exposure dose of the organs among the different plans were compared. Results There was no significant difference in the doses of the organs at risk (class I) under the premise of target dose requirements (all P>0.05). The increase of the weight of the target area and the organs at risk will increase the total MUs. The increase of the preset limit value of the minimum MUs in the subfield will reduce the total MUs. The increase of the fluence smoothness in the X and Y directions will increase the total MUs. An unreasonable minimum MU value will increase the total MUs. Under the condition that the organ exposure is not changed significantly, the influencing factors of MU are ranked as weight>fluence smoothness>minimum MUs. Conclusions Parameter setting and process planning can reduce the total MUs to a certain extent. However, due to the complexity of the influence of optimization parameters on the plan, the optimization process should be preferred. Especially in the Eclipse TPS, the method of gradual optimization to achieve the final dose distribution requirement and then remove the fluence re-optimization is more convenient and effective for reducing the total MUs.

No abstract available.
Diffusion Tensor Imaging ; Diffusion ; Hemiplegia ; Humans

OBJECTIVE: To investigate the clinical significance of bone marrow microvessel density(MVD) and angiogenesis related factors in multipic myeloma(MM). METHODS: Twenty cases of MM and 20 cases of simple fracture were selected and enrolled in MM group and control group respectively. The clinical data and results of laboratorial tests were collected; the bone marrow MVD of patients was detected by using the modified plastic-embedded pathologic sections of bone marrow tissue and histochemistry staining, the expression levels of amgiogenesis-related factors including VEGF, TNF-α, HGF, TGF-α, TGF-β1, bFGF, Ang-Ⅰ, Ang-Ⅱ in bone marrow supernatant were detected by ELISA; the mRNA expression levels of above-mentioned cytokines in bone marrow mononuclear cells were detected by real time-PCR; the pearson correlation analysis was used to analyze the correlation of MVD with VEGF, HGF and bFGF levels. RESULTS: The MVD in MM group was significantly higher than that in control group (P＜0.001); the mRNA expression of VEGF, TGF-α, TGF-β1 and HGF in bone marrow mononuclear cells of MM group was higher than that of control group(P＜0.001); the levels of VEGF, HGF, bFGF and THF-α in bone marrow supernatant of MM group were higher than those in control group(P＜0.05), moreover, the MVD positively correlated with levels of VEGF, HGF and bFGF in bone marrow(r=0.488, 0.472 and 0.457). CONCLUSION The MVD and levels vessel-related factors in bone marrow supernatant of MM patients increase, among which the levels of VEGF and HGF in bone marrow supernatant are consistant with those mRNA expression level in bone marrow mononuclear cells, moreover, the MVD possitively cerrelates with levels of VEGF, HGF and bFGF in bone marrow supernatant, suggesting that the changes of bone marrow microenvironment vassel-related factors play an important role in angiogenesis and pathogenesis of multiple myeloma.
Bone Marrow ; Bone Marrow Cells ; Humans ; Microvessels ; Multiple Myeloma ; Neovascularization, Pathologic ; Tumor Microenvironment

OBJECTIVE: To explore the pathogenesis of erythrocytosis by detecting the key enzymes of glucose metabolism and glucose transporter in bone marrow erythrocytes of chronic mountain sickness (CMS), and analyzing its correlation with hemoglobin. METHODS: Twenty CMS patients hospitalized in Qinghai Provincial People's Hospital from January 2019 to December 2020 were selected as CMS group. Twenty males with leukocyte count > 3.5×10⁹/L who had accepted bone marrow aspiration and had normal result were taken as control group. The mRNA and protein expression of key enzymes and glucose transporter in glucose metabolism in bone marrow CD71⁺ erythrocytes were detected by real time qPCR and Western blot, respectively. Glucose, lactic acid and 2,3-diphosphoglycerate in the bone marrow supernatant and serum were tested by ELISA. The mRNA and protein expression of key enzymes and glucose transporter, glucose, lactic acid and 2,3-diphosphoglycerate of the two groups were compared. Pearson correlation was used to analyze the correlation between key enzymes, glucose transporter in glucose metabolism in bone marrow CD71⁺ erythrocytes and hemoglobin. RESULTS: The expression of HK2, GLUT1 and GLUT2 mRNA in the CMS group were higher than those in the control group (P<0.001), while the expression of HK1, OGDH and COX5B mRNA were not different. The expression of HK2, GLUT1 and GLUT2 protein in the CMS group were higher than those in the control group (P<0.05). The levels of glucose and lactic acid in the bone marrow supernatant and serum in the CMS group were not different from those in the control group, while the level of 2,3-diphosphoglycerate was higher (P<0.001). Both HK2 and GLUT2 proteins were positively correlated with hemoglobin (r=0.511, 0.717). CONCLUSION CMS patients may increase glycolysis by increasing the expression of HK2, and promote the utilization of glucose through high expression of GLUT1 and GLUT2 to meet the need of energy supply.
Male ; Humans ; Altitude Sickness/metabolism* ; Glucose Transporter Type 1 ; 2,3-Diphosphoglycerate ; Hemoglobins ; Chronic Disease ; RNA, Messenger ; Phenotype ; Glucose

Objective: To investigate the effects of exosomes from human adipose-derived mesenchymal stem cells (ADSCs) on inflammatory response of mouse RAW264.7 cells and wound healing of full-thickness skin defects in mice. Methods: The experimental research methods were adopted. The discarded adipose tissue was collected from 3 female patients (aged 10-25 years) who underwent abdominal surgery in the First Affiliated Hospital of Air Force Medical University. ADSCs were extracted from the adipose tissue by collagenase Ⅰ digestion and identified with flow cytometry. Exosomes were extracted from the human ADSCs by differential ultracentrifugation, the morphology of the exosomes was observed by transmission electron microscopy, the particle diameter of the exosomes was detected by nanoparticle tracking analyzer, and the protein expressions of CD9, CD63, tumor susceptibility gene 101 (TSG101), and β-actin were detected by Western blotting. The human ADSCs exosomes (ADSCs-Exos) and RAW264.7 cells were co-cultured for 12 h, and the uptake of RAW264.7 cells for human ADSCs-Exos was observed. The RAW264.7 cells were divided into phosphate buffer solution (PBS) group stimulated with PBS for suitable time, endotoxin/lipopolysaccharide (LPS) stimulation 2 h group, LPS stimulation 4 h group, LPS stimulation 6 h group, LPS stimulation 12 h group, and LPS stimulation 24 h group stimulated with LPS for corresponding time, with 3 wells in each group, and the mRNA expressions of interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), IL-6, and IL-10 were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) method. The RAW264.7 cells were divided into PBS group, LPS alone group, and LPS+ADSCs-Exos group, with 3 wells in each group, which were dealt correspondingly for the time screened out in the previous experiment, the mRNA expressions of IL-1β, TNF-α, IL-6, IL-10, trasforming growth factor β (TGF-β,) and vascular endothelial growth factor (VEGF) were detected by real time fluorescence quantitative RT-PCR method, and the protein expressions of inducible nitric oxide synthase (iNOS) and arginase 1 (Arg1) were detected by Western blotting. Twenty-four 8-week-old male BALB/c mice were divided into PBS group and ADSCs-Exos group according to the random number table, with 12 mice in each group, and a full-thickness skin defect wound with area of 1 cm×1 cm was inflicted on the back of each mouse. Immediately after injury, the wounds of mice in the two groups were dealt correspondingly. On post injury day (PID) 1, the concentration of IL-1β and TNF-α in serum were detected by enzyme-linked immunosorbent assay, and the mRNA expressions of IL-1β, TNF-α, and IL-6 were detected by real time fluorescence quantitative RT-PCR method. On PID 3, 6, 9, 12, and 15, the wound healing was observed and the wound non-healing rate was calculated. On PID 15, the defect length of skin accessory and collagen volume fraction (CVF) were detected by hematoxylin eosin staining and Masson staining, respectively, the CD31 expression and neovascularization were detected by immunohistochemistry, and the ratio of Ki67 positive cells, the ratio of iNOS and Arg1 double positive cells, and the ratio of iNOS positive cells to Arg1 positive cells and their fluorescence intensities were detected by immunofluorescence method. The number of samples in animal experiments was 6. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and independent sample t test. Results: At 12 h of culture, the cells exhibited a typical spindle shape, which were verified as ADSCs with flow cytometry. The exosomes with a vesicular structure and particle diameters of 29-178 nm, were positively expressed CD9, CD63, and TSG101 and negatively expressed β-actin. After 12 h of co-culture, the human ADSCs-Exos were endocytosed into the cytoplasm by RAW264.7 cells. The mRNA expressions of IL-1β, TNF-α, IL-6, and IL-10 of RAW264.7 cells in LPS stimulation 2 h group, LPS stimulation 4 h group, LPS stimulation 6 h group, LPS stimulation 12 h group, and LPS stimulation 24 h group were significantly higher than those in PBS group (with t) values of 39.10, 14.55, 28.80, 4.74, 48.80, 22.97, 13.25, 36.34, 23.12, 18.71, 29.19, 41.08, 11.68, 18.06, 8.54, 43.45, 62.31, 22.52, 21.51, and 37.13, respectively, P<0.01). The stimulation 12 h with significant expressions of all the inflammatory factors was selected as the time point in the following experiment. After stimulation of 12 h, the mRNA expressions of IL-1β, TNF-α, IL-6, and IL-10 of RAW264.7 cells in LPS alone group were significantly higher than those in PBS group (with t values of 44.20, 51.26, 14.71, and 8.54, respectively, P<0.01); the mRNA expressions of IL-1β, TNF-α, and IL-6 of RAW264.7 cells in LPS+ADSCs-Exos group were significantly lower than those in LPS alone group (with t values of 22.89, 25.51, and 8.03, respectively, P<0.01), while the mRNA expressions of IL-10, TGF-β, and VEGF were significantly higher than those in LPS alone group (with t values of 9.89, 13.12, and 7.14, respectively, P<0.01). After stimulation of 12 h, the protein expression of iNOS of RAW264.7 cells in LPS alone group was significantly higher than that in PBS group and LPS+ADSCs-Exos group, respectively (with t values of 11.20 and 5.06, respectively, P<0.05 or P<0.01), and the protein expression of Arg1 was significantly lower than that in LPS+ADSCs-Exos group (t=15.01, P<0.01). On PID 1, the serum concentrations of IL-1β and TNF-α and the mRNA expressions of IL-1β, TNF-α, and IL-6 in wound tissue of mice in ADSCs-Exos group were significantly those in lower than PBS group (with t values of 15.44, 12.24, 9.24, 7.12, and 10.62, respectively, P<0.01). On PID 3, 6, 9, 12, and 15 d, the wound non-healing rates of mice in ADSCs-Exos group were (73.2±4.1)%, (53.8±3.8)%, (42.1±5.1)%, (24.1±2.8)%, and 0, which were significantly lower than (82.5±3.8)%, (71.2±4.6)%, (52.9±4.1)%, (41.5±3.6)%, and (14.8±2.5)% in PBS group, respectively (with t values of 4.77, 8.93, 5.54, 7.63, and 7.59, respectively, P<0.01). On PID 15, the defect length of skin accessory in wounds of mice in PBS group was significantly longer than that in ADSCs-Exos group (t=9.50, P<0.01), and the CVF was significantly lower than that in ADSCs-Exos group (t=9.15, P<0.01). On PID 15, the CD31 expression and the number of new blood vessels (t=12.99, P<0.01), in wound tissue of mice in ADSCs-Exos group were significantly more than those in PBS group, and the ratio of Ki67 positive cells was significantly higher than that in PBS group (t=7.52, P<0.01). On PID 15, the ratio of iNOS and Arg1 double positive cells in wound tissue of mice in PBS group was (12.33±1.97)%, which was significantly higher than (1.78±0.29)% in ADSCs-Exos group (t=13.04, P<0.01), the ratio of iNOS positive cells and the fluorescence intensity of iNOS were obviously higher than those of ADSCs-Exos group, and the ratio of Arg1 positive cells and the fluorescence intensity of Arg1 were obviously lower than those of ADSCs-Exos group. Conclusions: The human ADSCs-Exos can alleviate inflammatory response of mouse RAW264.7 cells, decrease macrophage infiltration and secretion of the pro-inflammatory cytokines, increase the secretion of anti-inflammatory cytokines to promote neovascularization and cell proliferation in full-thickness skin defect wounds of mice, hence accelerating wound healing.
Animals ; Exosomes ; Female ; Humans ; Male ; Mesenchymal Stem Cells ; Mice ; Skin ; Vascular Endothelial Growth Factor A ; Wound Healing