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Objective:To investigate the inhibitory effect of Sangsuyin on airway inflammation in asthmatic rats by regulating the Toll-like receptor 4 （TLR4） pathway. Method:Forty-eight SD rats were randomly divided into a normal group （<italic>n</italic>=8） and an experimental group （<italic>n</italic>=40）. Asthma model was induced in the rats of the experimental group which were further divided into the following five groups according to a random number table： a model group， a dexamethasone group （0.005 g·kg<sup>-1</sup>），and low- （2.1 g·kg<sup>-1</sup>）， medium- （4.2 g·kg<sup>-1</sup>）， and high-dose （8.4 g·kg<sup>-1</sup>） Sangsuyin groups. The drugs were all dissolved in normal saline at 0.01 L·kg<sup>-1</sup>. The rats in the model group and the normal group received normal saline （<italic>ig</italic>） at 0.01 L·kg<sup>-1</sup>. The drug treatment was carried out once per day， for a total of 7 days. The grades of allergic reactions were compared among the groups after intervention. The levels of interleukin-1<italic>β</italic> （IL-1<italic>β</italic>），tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）， and interferon-<italic>γ</italic>（IFN-<italic>γ</italic>） in serum and bronchoalveolar lavage fluid （BALF） were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of lung tissues，and the inflammatory cell infiltration scores of lung tissues were compared. The mRNA expression of TLR4 and nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） was detected by quantitative real-time fluorescence-based reverse-transcription polymerase chain reaction （Real-time PCR）. The expression of TLR4，NF-<italic>κ</italic>B， and the level of phosphorylated NF-<italic>κ</italic>B （p-NF-<italic>κ</italic>B） were detected by Western blot. Result:The success rate of modeling was 95.00%. Compared with the normal group， the model group showed a higher grade of allergic reaction and more severe pathological changes of lung tissues，and the groups with drug intervention exhibited relieved conditions. The levels of IL-1<italic>β</italic> and TNF-<italic>α</italic> in the serum and BALF were higher in the model group than in the normal group，and lower in the groups with drug intervention than in the model group （<italic>P</italic><0.01）. The level of IFN-<italic>γ</italic> in the serum and BALF was lower in the model group than in the normal group，and higher in the groups with drug intervention than in the model group （<italic>P</italic><0.05，<italic>P</italic><0.01）. The inflammatory cell infiltration score，mRNA and protein expression of TLR4 and NF-<italic>κ</italic>B，and the mRNA level of p-NF-<italic>κ</italic>B were higher in the model group than in the normal group，and lower in the groups with drug intervention than in the model group （<italic>P</italic><0.01）. Conclusion:Sangsuyin could inhibit allergic reactions，lung tissue lesions， and airway inflammation in asthmatic rats. It is speculated that this effect is achieved by inhibiting the TLR4 pathway，down-regulating the mRNA and protein expression of TLR4 and NF-<italic>κ</italic>B，and reducing the level of p-NF-<italic>κ</italic>B.angsuyin can inhibit allergic reaction，lung tissue lesions and airway inflammation in asthmatic rats. It is speculated that this effect is achieved by inhibiting TLR4 pathway，down regulating the mRNA and protein expression of TLR4，NF-<italic>κ</italic>B，and reducing the levels of p-NF-<italic>κ</italic>B.
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BACKGROUND: Studies have shown that osteoprotegerin/receptor activator of nuclear factor κB ligand/nuclear factor κB receptor activator (OPG/RANKL/RANK) signaling pathway has a certain correlation with the pathogenesis of giant-cell tumor. Controlling the OPG/RANKL/RANK signaling pathway to affect the interaction between osteoblasts and osteoclasts can play a certain therapeutic role in giant-cell tumor of bone. OBJECTIVE: Ton introduce the relationship between the OPG/RANKL/RANK signaling pathway and the pathogenesis of giant-cell tumor of bone, and to summarize and discuss the new advances of the OPG/RANKL/RANK signaling pathway in the pathogenesis of giant-cell tumor of bone. METHODS: PubMed, Web of Science, and WanFang databases were searched for relevant articles published from 2001 to 2019 using the keywords of “OPG/RANKL/RANK, giant cell tumor of bone, pathogenesis, signal pathway, bone metabolism" in English and Chinese, respectively. A total of 53 articles were finally included for analysis and discussion after removal of old and repeated literatures. RESULTS AND CONCLUSION: OPG inhibits the proliferation and differentiation of osteoclasts, reduces the activity of mature osteoclasts, and blocks the binding of RANKL to RANK. RANKL binds to RANK on the surface of osteoclast progenitor cells to promote the differentiation and proliferation of osteoclast progenitor cells, thus accelerating osteoclast progression. After binding to RANKL receptor, RANKL activates signal factors such as nuclear factor-κB to promote the proliferation, differentiation and activation of osteoclasts, and to regulate the transcription and expression of related genes. Therefore, the OPG/RANKL/RANK is associated with the pathogenesis of giant-cell tumor.
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Objective:To explore the effect of gymnastic exercise on sensory integration disorder (SID) in children aged three to six years. Methods:From March to June 2018, 27 children aged three to six years with SID were recruited from Chengdu U-Beller International Children Education Center (Pidu Campus), and were randomly divided into control group (n = 13) and experimental group (n = 14). The experimental group accepted gymnastic exercise, 60 minutes a time, three times a week, for 16 weeks. The control group received no intervention. They were assessed with Assessment Scale for Children Sensory Integration Development, for vestibular dysfunction, tactile defense and proprioception dysfunction. Results:After intervention, the SID improvement was better in the experimental group than in the control group (χ2 > 6.639, P < 0.05), several children with mild disorder returned to normal level, and several with severe disorder returned to mild level. There was no difference in the scores of vestibular dysfunction, tactile defense and proprioception dysfunction before and after intervention in the control group (P > 0.05), and the scores significantly improved in the experimental group (|t| > 7.015, P < 0.01), and was higher in the experimental group than in the control group after intervention (t > 2.193, P < 0.01). Conclusion:Gymnastic exercise can improve vestibular dysfunction, tactile defense and proprioception dysfunction for children with SID aged three to six years.
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Objective: To investigate the safety and efficacy of regional transport to percutaneous coronary intervention(PCI) hospitals from non-PCI hospitals after thrombolysis in patients with acute ST-segment elevation myocardial infarction(STEMI) in northwest China. Methods: In this retrospective study, 1 062 STEMI patients who were transferred from non-PCI hospitals within 24 hours from symptom onset, during January 2015 and January 2019 in the First Hospital of Lanzhou University, were included. According to the treatment strategy, they were divided into two groups, namely intravenous thrombolysis combined with PCI group(n=240), and primary PCI group(n=822). Observation endpoint were in-hospital adverse cardiovascular and cerebrovascular events and bleeding events, Including all-cause death, ischemic stroke, malignant arrhythmia, intracranial hemorrhage and hemorrhage with hemoglobin decrease≥50 g/L. Results: A total of 1 062 STEMI patients were included(age was (61±12) years old), with 905 males (85.2%). The proportion of grade 0 TIMI blood flow in the primary PCI group before operation was significantly higher than that in the thrombolysis combined with PCI group(63.0%(518/822) vs. 36.3%(87/240)， P<0.001). Compared with primary PCI group, the time from symptom onset to first medical contact(2.11(1.00, 4.00)hours vs.3.00(1.13, 7.07)hours, P<0.001) and reperfusion in thrombolysis combined with PCI group(3.07(1.83, 4.87)hours vs. 6.92(4.07, 11.15) hours, P<0.001) were significantly shorter. The proportion of all-cause death was significantly higher in the primary PCI group than that in the thrombolysis combined with PCI group (1.8%(15/822) vs. 0, P=0.03). There was no significant difference in hemorrhage, ischemic stroke and malignant arrhythmia between the two groups(all P>0.05). Conclusions: For STEMI patients initially hospitalized in non-PCI hospitals, regional transport combined with PCI is feasible and effective. It does not significantly increase the risk of bleeding and cardiovascular and cerebrovascular events, with shorter time from symptom onset to myocardial reperfusion.
Subject(s)
Aged , Humans , Male , Middle Aged , Angioplasty, Balloon, Coronary , China , Myocardial Infarction , Percutaneous Coronary Intervention , Retrospective Studies , ST Elevation Myocardial Infarction/therapy , Thrombolytic Therapy , Treatment OutcomeABSTRACT
Objective: To study the relationship between plasma levels of Neutrophil gelatinase- associated lipocalin (NGAL), matrix metalloproteinase (MMP-9), hs-CRP, IL-1β and the severity of coronary stenosis in acute ST-elevation myocardial infarction (STEMI) patients without kidney disease; to explore the ability for NGAL discriminating severe coronary stenosis. Methods: Our research included in 2 groups: STEMI group, n=124 patients admitted in our hospital from 2014-01 to 2017-03 and Control group, n=124 subjects without obvious coronary stenosis. According to SYNTAX score, the patients were divided into another set of 2 groups: High score group, patients with SYNTAX score ≥ 33, n=26 and Low score group, patients with SYNTAX score < 33, n = 98. Relationship between plasma levels of NGAL, MMP-9, hs-CRP, IL-1β and the severity of coronary stenosis were studied by Spearman correlation analysis; the ability for NGAL discriminating severe coronary stenosis was examined by ORC curve; the influencing factors for SYNTAX score were determined by Logistic regression analysis. Results: Compared with Control group, STEMI group had increased plasma levels of NGAL, MMP-9, hs-CRP, all P < 0.01,while IL-1β was similar between 2 groups, P=0.272. NGAL was positively related to MMP-9 (r=0.757, P<0.01), SYNTAX score (r=0.616, P<0.01) and IL-1β (r=0.185, P<0.05). Compared with Low score group, High score group showed the higher plasma levels of NGAL, MMP-9, both P<0.01, while the lower LVEF, P<0.01. The area under ROC curve for NGAL discriminating severe coronary stenosis was 0.881 (95% Cl 0.813-0.949, P<0.01) which was greater than MMP-9 [0.799 (95% Cl 0.709-0.890, P<0.01)] and hs-CRP [0.446 (95% Cl 0.306-0.587, P=0.400)]. Multivariate regression analysis presented that plasma NGAL level was independently related to high SYNTAX score [OR=1.115, 95% Cl (1.107-1.123), P<0.01]. Conclusion: STEMI patients had increased plasma levels of NGAL, MMP-9 and hs-CRP; NGAL had better ability for discriminating severe coronary stenosis than MMP-9 and hs-CRP. NGAL as a new biomarker should be helpful for risk stratification in STEMI patients.
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Objective To evaluate the safety and efficacy of continuous renal replacement therapy (CRRT) during percutaneous coronary intervention (PCI) in patients with severe acute myocardial infarction (AMI). Methods A total of 945 patients with AMI who were hospitalized in the CCU of the First Hospital of Lanzhou University from January 2016 to December 2017 were retrospectively analyzed and 21 patients had undergone perioperative CRRT during percutaneous coronary intervention. Baseline clinical characteristics of the AMI patients were evaluated before and after CRRT treatment. The changes in heart function, renal function, liver function and other parameters were evaluated. Results The heart rate of patients receiving CRRT was significantly lower at 24h and 48h after CRRT than that of before treatment (P=0.038). Decrease in mean systolic blood pressure and diastolic blood pressure was observed af ter CRRT treatment without statistical significance (P>0.05). Proportion of patients with Killip class I ~ II heart function increased significantly after CRRT[23.8%(5/21)vs. 57.1%(12/21),P=0.001]. Amelioration in urea nitrogen, creatinine, aspartate aminotransferase, glutamic pyruvic transaminase and total bilirubin were found at 24h and 48h after CRRT treatment compared to pre-CRRT levels (P<0.05). Conclusions Perioperative CRRT is safe and effective for patients with severe AMI.
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This study was purposed to construct the recombinant hF9 minigene and its stable nonsense mutant cell lines, and to investigate its significance. Minigene hF9 was cloned into the mammalian expression vector pCMV-Tag3B; a nonsense mutant containing a premature termination codon (PTC) in the 121(st) amino acid residue was obtained by PCR site-directed mutagenesis; minigene hF9 and nonsense mutant were respectively transfected into HepG2 cells with G418 treatment to get stable HepG2-WT and HepG2-N cell lines. The results confirmed that the minigene hF9 and nonsense mutant were constructed successfully. The gene of interest was amplified by RT-PCR from the stable cell lines, and the minigene hF9 was expressed in the stable cell lines. It is concluded that the recombinant hF9 minigene and its stable nonsense mutant cell lines are constructed successfully. The cell lines can be used to screen the drugs treating the nonsense mutation-caused hemophilia according to PTC read-through approaches.
Subject(s)
Humans , Cell Line, Tumor , Codon, Nonsense , Factor IX , Genetics , Genetic Vectors , Hemophilia B , Genetics , Recombinant Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the feasibility of transfecting recombinant Sp1 into hypertrophic scar fibroblasts and investigate the proliferation and collagen I, III synthesis in the transfected cells.</p><p><b>METHODS</b>Recombinant human Sp1 was transfected into hypertrophic scar fibroblasts with the karyocyte expressive vector. The expression of Sp1, collagen I, III mRNA was tested by real time PCR. The change of cell proliferation was observed with CCK8 colorimeter.</p><p><b>RESULTS</b>About 30% of transfected hypertrophic scar fibroblasts showed green fluorescence positive. The relative expression of Sp1 mRNA in transfected cells, empty-vector cell or untransfected cells group was 5.26 +/- 0.76, 1.08 +/- 0.18, 1.09 +/- 0.15, respectively, showing a significant difference between thansfected and untransfected cells or between the transfected cells and empty-vector group (P <0.01, n = 5). Expression of collagen I, III mRNA was 2.49 +/- 0.40 and 1.88 +/- 0.30 in transfected cells, 0.96 +/- 0.18 and 0.95 +/- 0.18 in empty-vector cell, and 0.97 +/- 0.15 and 0.93 +/- 0.13 in untransfected cells, respectively, showing a significant difference between thansfected and untransfected cells or between the transfected cells and empty-vector group (P < 0.01, n = 5).</p><p><b>CONCLUSIONS</b>The hypertrophic scar fibroblasts could be as the target cells of Sp1 gene transfection. Sp1 gene may play an important role in abnormal collagen metabolism in hypertrophic scar.</p>
Subject(s)
Humans , Cell Proliferation , Cells, Cultured , Cicatrix, Hypertrophic , Genetics , Metabolism , Pathology , Collagen , Metabolism , Escherichia coli , Genetics , Fibroblasts , Metabolism , Pathology , RNA, Messenger , Genetics , Recombinant Proteins , Genetics , Skin , Metabolism , Sp1 Transcription Factor , Genetics , TransfectionABSTRACT
<p><b>AIM</b>To investigate the transformation of characteristics of epidermal cells from foreskin which were used to reconstruct male rabbit anterior urethra in combination with acellular collagen matrices.</p><p><b>METHODS</b>In three rabbits, autologous foreskin epidermal cells were isolated, expanded in vitro, and seeded (inoculated) onto a tubular acellular collagen matrix, acquired from allogeneic rabbit bladder submucosa. A urethral mucosal defect was created, and urethral reconstruction was performed with the tubular acellular collagen matrix seeded with epidermal cells.</p><p><b>RESULTS</b>On gross examination at 12 months following the procedure, the mucosa of the urethral grafts appeared lubricous and smooth. Urethrography showed that a wide urethral caliber had been maintained without any sign of strictures. Histological examination showed a transitional cell layer in the graft without evidence of a margin between the graft and the host tissue at 12 months postoperatively.</p><p><b>CONCLUSION</b>Epidermal cells seeded onto acellular collagen matrices can be successfully used to reconstruct urethras that have defects and are transformed to transitional epithelial cells.</p>
Subject(s)
Animals , Male , Rabbits , Cell Transplantation , Methods , Collagen , Epidermis , Cell Biology , Foreskin , Cell Biology , Graft Survival , Mucous Membrane , Cell Biology , Plastic Surgery Procedures , Methods , Tissue Culture Techniques , Tissue Engineering , Methods , Urethra , General Surgery , Urethral Stricture , General SurgeryABSTRACT
<p><b>UNLABELLED</b>[Abstract]</p><p><b>OBJECTIVE</b>The study was (1) to investigate the biological character changes of human epidermal cells during proliferation culture in vitro and (2) to provide data for construction of engineered skin.</p><p><b>METHODS</b>The foreskin was collected from 20 healthy children. The epidermal cells were isolated with digestion of the foreskin and cultured in vitro. Growth curve was obtained from the data of cell counting. Cell growth kinetics was observed. Meanwhile, clonal analysis and cell size measurement was performed. The rate of keratin 19 (K19) and involucrin expression-positive cells was counted by flow cytometer. Expression of K19 and involucrin mRNA was detected by RT-PCR.</p><p><b>RESULTS</b>When passaged to P6, the keratinocytes from child foreskin could be expanded by (700 +/- 37) times. Flow cytometer results showed that the percentage of K19 expression-positive cells decreased from (66.97 +/- 3.14)% to (4.65 +/- 1.38)% while the percentage of involucrin expression-positive cells increased from (11.65 +/- 1.62)% to (97.03 +/- 2.66)% at P0 and P6 respectively. RT-PCR results showed that expression of K19 mRNA decreased from P0 to P6 while involucrin mRNA kept stable with passage in vitro.</p><p><b>CONCLUSIONS</b>Human epidermal cells of passage 5 maintain proliferation phenotype, which are suitable for skin tissue engineering. Decrease of proliferation phenotype content is partially responsible for the proliferation capacity loss of in vitro cultured epidermal cells.</p>