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Objective: To investigate the main mechanisms of pulmonary fibrosis following silica nanoparticles (SiNPs) exposure through constructing the macrophage-fibroblast model in vitro, which simulated the process of pulmonary fibrosis. Methods: In January 2021, human mononuclear leukemia cells (THP-1) were treated with 0, 25, 50, 100 μg/ml SiNPs for 24 h. The supernatant of THP-1 cells was collected and applied to human embryonic lung fibroblast cells (MRC-5) which divided into control and low, medium and high dose groups at the logarithmic growth stage for 24 h. MRC-5 cell viability was detected by CCK8. The hydroxyproline (Hyp), interleukin 6 (IL-6), interleukin 1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) expression were detected in the supernatants of MRC-5. The changed proteins were detected by liquid-phase mass spectrometry in high dose group. GeneCard database were applied to identity the differential pulmonary fibrosis proteins in high dose group. Gene Ontology (GO) was performed to identity the key biological process in differential pulmonary fibrosis proteins of high dose group. The String database was used to construct the protein-protein interactions (PPI) network of differential pulmonary fibrosis proteins. The APP of CytoHubba was applied to calculate the key protein of differential pulmonary fibrosis proteins in PPI network. Correlation coefficients between key differential pulmonary fibrosis proteins were calculated using Pearson correlation analysis. Western blotting was applied to detect the expression of key proteins of differential pulmonary fibrosis proteins in different groups. Results: CCK8 results showed that MRC-5 cell viability was increasing in low, medium and high dose groups compared with control group (P<0.05). The expression levels of Hyp and IL-1β in different group were increased compared with control group, the expression levels of IL-6 and TNF-α were increased in high dose group compared with control group (P<0.05). GeneCard database identified 26 differential pulmonary fibrosis proteins, which were mainly involved in extracellular matrix hydrolysis, cell inflammatory response, tissue repair, cell proliferation, inflammation response by GO analysis. The APP of CytoHubba was calculated that matrix metalloproteinase 9 (MMP9) and tissue inhibitor metalloproteinase 1 (TIMP1) played an important role in PPI network. The results of correlation analysis showed that MMP9 was correlated with the expression of matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 3 (MMP3), TIMP1 and epidermal growth factor receptor (EGFR) (r=0.97, 0.98, 0.94, 0.93, P<0.05). Western blotting results showed that TIMP1 protein expression was increased in low, medium and high dose groups, while MMP9 protein expression was increased only in high dose group (P<0.05) . Conclusion: Differential expression proteins related with pulmonary fibrosis in MRC-5 cells mainly regulate biological processes of extracellular matrix hydrolysis, tissue repair, and cellular inflammation response following SiNPs exposure. MMP9 and TIMP1 may be the key proteins, which affected the fibrosis process in vitro pulmonary fibrosis model.
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Objective:To investigate the effect of long non-coding BANCR on the migration and invasion of melanoma cells and its mechanism.Methods:qPCR was used to detect the expression of BANCR in melanoma patients and the relationship between clini-copathological data.Kaplan-Meier analysis the survival of melanoma patients with different expression of BANCR.Transwell invasion assay was used to detect the effect of BANCR on the invasive ability of melanoma cells.The effect of BANCR on the migration of melanoma cells was detected by scratch healing assay.Western blot was used to detect Wnt/β-catenin Signal pathway protein expression.Results:BANCR was highly expressed in melanoma,especially with the higher pathological stage or the lymph node metastasis.The higher expression of BANCR,the worse survival of melanoma patients.The inhibition of BANCR expression could reduce the invasion and migration ability of melanoma cells.The expression of β-catenin and c-myc protein in Wnt/β-catenin signaling pathway was down-regulated after silencing BANCR.Conclusion:Long non-coding BANCR was highly expressed in melanoma patients and was negatively correlated with survival time,it also regulates melanoma cell migration and invasion by activating the Wnt/β-catenin signaling pathway.
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Background Diabetic retinopathy (DR) is one of the most important microvascular complications of diabetes,which has become one of the leading causes of blindness.Neovascularization is the main pathological manifestations of DR,but its mechanism is unknown.There is a clear need to investigate its pathogenesis which can offer potential therapeutic targets.Objective The aim of this study was to investigate the expression and distribution of visfatin and vascular endothelial growth factor (VEGF) in diabetic model rats.Methods This study was approved by Animal Ethic Committee of Inner Mongolia Medical University.Sixty SPF 8-week-old male SpragueDawley rats were randomized into the diabetic group and control group.The rats were housed under a condition that alternated between 12 hours of light and darkness,with free access to rat food and water.Diabetes was induced by intraperitoneal injection of 60 mg/kg (0.60 ml/100 g) of streptozotocin (STZ) and control rats received equivalent volume of buffer.The models were regarded as successful when blood glucose was ≥ 16.7 mmol/L.Rats were sacrificed 12 weeks after the injection of STZ and retinal specimens were prepared to detect the expression of visfatin and VEGF.Total retinal protein was isolated from the retinas of experimental and control eyes,and the expression of visfatin and VEGF was assessed by Western blot.Frozen cross sections of retinas of 5 μm thickness were used to perform double immunofluorescence staining with anti-visfatin and anti-VEGF antibodies.Results Mean body weight of the diabetic rats was (189.02±11.34) g and that of the control rats was (489.57 ± 14.48) g at 12 weeks post-injection,showing a significant difference between them (t =5.236,P =0.003).Mean blood glucose level was (29.25±3.86) mmol/L in the diabetic group and (5.32±1.01) mmol/L in the control group,demonstrating a significant difference (t =11.778,P =0.000).Double immunofluorescence staining showed reduced expression of visfatin and VEGF in the retinal nerve fibrous layer and glial cells in the control rats.A stronger staining for visfatin and VEGF was found in the various layers of the retina in the diabetic rats,with an expression level of visfatin (A value) of 346.26±41.23,which was considerably higher than that of the control group (102.07±65.01) (t =8.291,P =0.000) in 12 weeks after injection.Furthermore,the expression of VEGF in the retina was elevated in the diabetic group compared with the control group (A value) (415.88±92.15 vs.113.06±32.06) (t=10.067,P=0.000).Conclusions Visfatin might contribute to the pathologic progression of diabetic retinal,neovascularization and it might play a synergistic role with VEGF in the pathophysiology of DR.
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<p><b>BACKGROUND</b>Many types of human tumors can suppress the immune system to enhance their survival. Loss or down-regulation of human leukocyte antigens (HLA) class I on tumors is considered to be a major mechanism of tumor immune escape. Our previous studies found that HLA class I on peripheral-blood mononuclear cells was significantly lower in gastric cancer patients. The present study made an analysis of HLA class I expression on peripheral-blood T lymphocytes and NK cells from subjects of Lijiadian village, a village with high-incidence gastrointestinal tumor.</p><p><b>METHODS</b>A total of 181 villagers from Lijiadian village and 153 normal controls from the Department of Health Examination Center were enrolled in this study. Using a multi-tumor markers detection system, these villagers were divided into two groups: high-risk group (tumor markers positive group) and low-risk group (tumor markers negative group). The percentage of T lymphocytes and NK cells and levels of HLA class I on their surface were determined in these subjects by flow cytometry.</p><p><b>RESULTS</b>Percentages of T lymphocytes and NK cells in peripheral-blood mononuclear cells did not vary with age. The expression level of HLA class I on peripheral T lymphocytes and NK cells was not affected by age or gender, but was significantly down-regulated in Lijiadian villagers (P < 0.05), especially on the surface of NK cells (P < 0.01). Compared with the low-risk group, there was a significant reduction of HLA class I on peripheral T lymphocytes (P < 0.05) and NK cells (P < 0.05) in the high-risk group.</p><p><b>CONCLUSIONS</b>HLA class I on peripheral T lymphocytes and NK cells may be involved in tumorigenesis and development of gastrointestinal tumor, and understanding their changes in expression may provide new insights into the mechanism of tumor immunity.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Down-Regulation , Gastrointestinal Neoplasms , Allergy and Immunology , Histocompatibility Antigens Class I , Killer Cells, Natural , Allergy and Immunology , T-Lymphocytes , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the role of interleukin (IL)-18 and IL-18 receptor (IL-18R) in the predominant Th1 type cytokine response in patients with immune thrombocytopenia (ITP).</p><p><b>METHODS</b>Fifteen patients with active phase ITP, eighteen in remission and thirteen healthy controls were enrolled in this study. T-bet and GATA-3 mRNA levels in peripheral blood mononucleated cells (PBMNC) were measured by reverse transcriptase polymerase chain reaction (RT-PCR); the plasma IL-18 level by enzyme linked immunosorbent assay (ELISA), the expression of IL-18R on CD3(+) lymphocytes and total lymphocytes by flow cytometry(FCM).</p><p><b>RESULTS</b>The T-bet mRNA levels in patients with active phase ITP was 3.572 fold as much as that in the controls (P < 0.05), while the GATA-3 mRNA levels were 0.378 fold of that in controls (P < 0.05). The levels of plasma IL-18 and IL-18R on CD3(+) lymphocytes were significantly increased in active phase ITP than in remission phase and controls. There was no difference in ratio of T-bet/GATA-3 between remitted ITP and controls and so was for T-bet mRNA, GATA-3 mRNA, plasma IL-18 and IL-18R on CD3(+) lymphocytes.</p><p><b>CONCLUSION</b>ITP as a disease of Th1-dominant response there is an unbalance between T-bet and GATA-3 in its active phase; IL-18 and IL-18R being upregulated.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Case-Control Studies , GATA3 Transcription Factor , Metabolism , Interleukin-18 , Allergy and Immunology , Metabolism , Purpura, Thrombocytopenic, Idiopathic , Allergy and Immunology , Metabolism , Receptors, Interleukin-18 , Allergy and Immunology , Metabolism , T-Box Domain Proteins , Metabolism , Th1 Cells , Allergy and Immunology , MetabolismABSTRACT
Objective To investigate the changes of plasma resistin and lysophosphatidic acid and their roles in cerebral infarction. Methods Enzyme-linked immunosorbent assay (ELISA) was used to measure the plasma resistin levels in 130 patients with cerebral infarction, 40 with other neurological diseases (20 with sciatica and 20 with trigeminal neuralgia), and 60 healthy control subjects. Lysophosphatidic acid was isolated from the blood samples from the subjects and quantified by assaying its phosphorous component. Results The plasma levels of resistin and lysophosphatidic acid were significantly higher in patients with cerebral infarction than those in the other two groups (P<0.05). In patients with cerebral infarction, higher plasma resistin and plasma lysophosphatidic acid levels were associated with increased infarct size, and their levels were also found to closely correlate to the neurological deficits in the patients. Conclusion Detection of the plasma levels of resistin and lysophosphatidic acid may help estimate the severity of cerebral infarction, the infarct size and the degree of neurological deficits.
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Objective To find out the status of brick-tea type fluorosis in the epidemic areas.Methods Based on "Scheme for Epidemiological Brick-tea Type Fluorosis in Sichuan Province",ten counties were selected in Sichuan brick-tea areas and ten towns were selected in every county,then the epidemicologic survey was performed in children of 8~12 year-old and adults aged above 20 years old.Results 5044 children and 4053 adults were selected from brick-tea areas.The rates of dental fluorosis in children and adults were 55.69%(2809/5044)and 60.41%(4053/6709)respectively.The dental fluorosis was mainly of mild damage.The skeletal fluorosis found in X-ray film was 44.64%(167/1241)and in clinical examination,38.94%(3883/9973).The levels of urine fluoride in children and adults were 1.88 and 2.78 mg/L.The level of urine fluoride was not differenet among children of different age,but in adults it was higher in the elder than the younger.The level of fluoride in urine was related to the severeness of skeletal fluorosis(r=0.74).The detective rates of skeletal fluorosis in agricuIture,pasturing,and agriculture-pasturing areaswere 31.70%(1369/4318),50.04%(1228/2454),and 40.17%(1286/3201),respectively.The X-ray detecting rates of skeletal fluorosis in men and wonlen were 49.57%(229/462)and 41.72%(325/779) respectively(χ2=11.72,P<0.05).Conclusion The prevalence of brick-tea type fluorosis is very serious in the regions studied.
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<p><b>OBJECTIVE</b>To investigate the relationship between the expression of host human leukocyte antigen-B mRNA (HLA-B mRNA) and HLA-B antigen in peripheral blood leukocytes (PBLs) and the differentiation and metastasis of gastric carcinoma (GC).</p><p><b>METHODS</b>To design and screen specific primers of HLA-B gene independently, detect the expression of HLA-B mRNA from 30 GC patients by reverse transcription-PCR and compare with the HLA-B antigen expression measured by flow cytometry.</p><p><b>RESULTS</b>The expression rate of PBL HLA-B mRNA from GC patients (23. 3% ) was very significantly lower than that of normals (87. 5% ) (P <0. 01) , especially concerning the poorly differentiated GC patients with lymph node metastasis (16. 0% ). Measured by flow cytometry, the expression percentage of HLA-B antigen of well-differentiated GC patients without lymph node metastasis was 88. 2% , an obviously decreasing tendency was showed in comparison with that in the normal group (98. 8% ) , although the difference was not significant (P = 0. 056) , and the expression percentage in poorly differentiated GC patients with lymph node metastasis(73. 3% )was declined significantly (P <0. 05).</p><p><b>CONCLUSION</b>The expression of PBL HLA-B mRNA and HLA-B antigen in GC patients is decreased or lost, and correlated with differentiation and metastasis of the cancer. The expression of PBL HLA-B mRNA may more directly reflect its relationship with the tumor differentiation and metastasis than that of HLA-B antigen.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Cell Differentiation , Flow Cytometry , Gene Expression Regulation, Neoplastic , HLA-B Antigens , Genetics , Leukocytes , Allergy and Immunology , Metabolism , Lymphatic Metastasis , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Stomach Neoplasms , Genetics , Allergy and Immunology , PathologyABSTRACT
<p><b>AIM</b>To explore the effects of sodium hydrogen exchanger (NHE-1) specific hammerhead ribozyme on the expression and activity of NHE-1 and pHi in pulmonary artery smooth muscle cells (PASMCs) in rats, and its role in PASMCs proliferation.</p><p><b>METHODS</b>According to the secondary structure of NHE-1 mRNA in rats, NHE-1 specific hammerhead ribozyme was designed with the assistance of computer. The recombinant vector of retroviral plasmid pLXSN and NHE-1 hammerhead ribozyme, PRZ, was transfected into the cultured PASMCs. G418 resistant cell clones were screened with 60 microg/ml G418. Then, the expression of NHE-1 mRNA was detected by RT-PCR, intracellular pH was measured with fluorescent probe-BCECF, 22Na and 3H-TdR incorporation were determined respectively.</p><p><b>RESULTS</b>Compared with the cells transfected with pLXSN and non-transfected cells, NHE-1 mRNA, pHi value, 3H-TdR and 22Na incorporation decreased significantly in cells transfected with recombinant vector PRZ. No significance was found between the pLXSN transfected group and non-transfected group.</p><p><b>CONCLUSION</b>NHE-1 hammerhead ribozyme can cleave the target RNA specifically, reduce the expression of NHE-1 mRNA, induce intracellular acidosis and consequently prohibit the proliferation of PASMCs.</p>
Subject(s)
Animals , Rats , Cell Proliferation , Muscle, Smooth, Vascular , Myocytes, Smooth Muscle , Cell Biology , Pulmonary Artery , Cell Biology , Pathology , RNA, Catalytic , Genetics , RNA, Messenger , Genetics , Sodium-Hydrogen Exchanger 1 , Sodium-Hydrogen Exchangers , Metabolism , TransfectionABSTRACT
Objective To assss the clinical significance of Cystatin C(Cys C)as a marker of renal function in kidney transplant patients especially when infection or acute rejection occured.Methods Among 65 renal transplant recipients the concentrations of serum Cys C and serum creatinine(Scr)were determined before and one month after the transplantation,and also in the day and next day of occurrence of infection or rejection.Meanwhile,30 healthy persons and 30 infected patients without kidney transplantations were served as control.Results The concentrations of Cys C were nearly equal between healthy persons and the infected patients without kidney transplantations(P = 0.32).The level of serum Cys C and Scr dropped quickly in the first 3 days after transplantation(decreased by 69.2%,74.7%,75.8% for Cys C and 38.4%,74.5%,81.4% for Scr)(P