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Objective:To explore the effect of high mobility group box-1 protein (HMGB1) on the balance of Th17/Treg in patients with immune thrombocytopenia (ITP).Methods:A total of 30 patients who were first diagnosed as ITP in the Fifth People's Hospital of Datong from July 2017 to April 2018 were selected as the case group, and another 30 healthy volunteers in the corresponding period were taken as the control group. The proportion of Th17 and Treg cells was detected by using flow cytometry, and the concentration of HMGB1, interleukin (IL)-17 and transforming growth factor β (TGF-β) in plasma was tested by using enzyme-linked immunosorbent assay (ELISA). Isolated peripheral blood mononuclear cells (PBMC) were cultured in vitro. After the treatment with recombinant human HMGB1 (rhHMGB1), real-time polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression changes in Treg cell transcription factor intracellular forkhead helix transcription factor 3 (Foxp3) and Th17 cell transcription factor retinoid related orphan receptor γt (RORγt). The differences of indicators in Treg cell transcription factor peripheral blood between the case group and the control group were compared, and the balance correlation between HMGB1 and Th17/Treg was analyzed.Results:Compared with the healthy control group, the proportion of Th17 cells and the expression level of HMGB1 and IL-17 in peripheral blood of ITP patients were increased (all P < 0.01), while the proportion of Treg cells and the level of TGF-β were decreased (all P < 0.01). The proportion of Th17 cells and the expression level of IL-17 and HMGB1 in peripheral blood of ITP patients were positively correlated with the concentration of HMGB1 (all P < 0.01); the proportion of Treg cells and the level of TGF-β were negatively correlated with the expression level of HMGB1 (all P < 0.01). In vitro experiments, the expression of intracellular RORγt mRNA was increased compared with the negative control group (1.50±0.24 vs. 0.93±0.22, t = 9.612, P < 0.01), and the expression of Foxp3 mRNA was decreased compared with the negative control group after the stimulation of PBMC by rhHMGB1 (0.72±0.19 vs. 1.08±0.18, t = 7.387, P < 0.01). Conclusion:The high level of HMGB1 in the peripheral blood of ITP patients induces Th17/Treg imbalance and aggravates inflammatory reactions, which may be an important cause of ITP.
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Objective@#To explore the effect of high mobility group box-1 protein (HMGB1) on the balance of Th17/Treg in patients with immune thrombocytopenia (ITP).@*Methods@#A total of 30 patients who were first diagnosed as ITP in the Fifth People's Hospital of Datong from July 2017 to April 2018 were selected as the case group, and another 30 healthy volunteers in the corresponding period were taken as the control group. The proportion of Th17 and Treg cells was detected by using flow cytometry, and the concentration of HMGB1, interleukin (IL)-17 and transforming growth factor β (TGF-β) in plasma was tested by using enzyme-linked immunosorbent assay (ELISA). Isolated peripheral blood mononuclear cells (PBMC) were cultured in vitro. After the treatment with recombinant human HMGB1 (rhHMGB1), real-time polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression changes in Treg cell transcription factor intracellular forkhead helix transcription factor 3 (Foxp3) and Th17 cell transcription factor retinoid related orphan receptor γt (RORγt). The differences of indicators in Treg cell transcription factor peripheral blood between the case group and the control group were compared, and the balance correlation between HMGB1 and Th17/Treg was analyzed.@*Results@#Compared with the healthy control group, the proportion of Th17 cells and the expression level of HMGB1 and IL-17 in peripheral blood of ITP patients were increased (all P < 0.01), while the proportion of Treg cells and the level of TGF-β were decreased (all P < 0.01). The proportion of Th17 cells and the expression level of IL-17 and HMGB1 in peripheral blood of ITP patients were positively correlated with the concentration of HMGB1 (all P < 0.01); the proportion of Treg cells and the level of TGF-β were negatively correlated with the expression level of HMGB1 (all P < 0.01). In vitro experiments, the expression of intracellular RORγt mRNA was increased compared with the negative control group (1.50±0.24 vs. 0.93±0.22, t = 9.612, P < 0.01), and the expression of Foxp3 mRNA was decreased compared with the negative control group after the stimulation of PBMC by rhHMGB1 (0.72±0.19 vs. 1.08±0.18, t = 7.387, P < 0.01).@*Conclusion@#The high level of HMGB1 in the peripheral blood of ITP patients induces Th17/Treg imbalance and aggravates inflammatory reactions, which may be an important cause of ITP.
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OBJECTIVE: To study the effects of ethanol extract of Sanguis Draconis on the survival of perforating flap model in rats and PI3K/Akt/eNOS pathway. METHODS: Perforating flap model was established by cutting off surrounding vessels and keeping one perforator. After modeling, the rats were divided into model group (external use, normal saline) and ethanol extract of Sanguis Draconis (EESD, the content of dracorhodin was 75.08 mg/g) group (external use, 0.21 g/cm2), with 10 rats in each group. They were given relevant medicine for consecutive 7 days, once a day. The flap survival rate and flap microvessel density were determined after given relevant medicine 7 days. Human umbilical vein endothelial cells (HUVECs) were reoxygenated and glycoconjugated 16 h after hypoxia and hypoglycemia to establish oxygen-glucose deprivation/oxygen-glucose recovery model of HUVECs. After modeling, model cells were divided into normal group, model group, dracorhodin high-concentration, medium- concentration and high-concentration groups (2.5, 1.0, 0.5 μg/mL). After reoxygenated and glycoconjugated for 24 h, cells morphology was observed by microscope; cell viability and the content of NO were detected by MTT assay and colorimetry. mRNA expression of Akt, PI3K and eNOS, PI3K protein expression, the phosphorylation of Akt and eNOS protein were determined by RT-PCR and Western blot assay. RESULTS: In rat experiment, compared with model group, flap survival rate and microvessel density of rats were increased significantly in EESD group (P<0.01). In cell experiment, compared with normal group, the survival rate of HUVEC, NO content, mRNA expression of PI3K, Akt, eNOS,PI3K protein expression, the phosphorylation of Akt and eNOS protein were decreased significantly (P<0.05 or P<0.01). Compared with model group, dracorhodin high-concentration, medium-concentration and high-concentration groups survival rate of HUVEC cells, NO content, mRNA expression of PI3K, Akt and eNOS, PI3K protein expression, the phosphorylation of Akt and eNOS protein were increased significantly (P<0.05 or P<0.01). CONCLUSIONS: The survival rate of perforating flap model in rat can be increased by treating with EESD, the mechanism of which may be associated with the activation of PI3K/Akt/eNOS pathway to protect endothelial cells.
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Inflammatory bowel disease (IBD)is an autoimmune disease leading to diarrhea,abdominal pain,and weight loss. The pathogenesis of diarrhea remains unclear,however,increasing evidence has shown that the epithelial sodium channel (ENaC)is associated with diarrhea. ENaC is crucial in the control of sodium homeostasis,extracellular fluid volume,blood pressure. This article reviewed advances in study on relationship between ENaC and IBD.
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Objective To investigate the effect of lateral ventricle transplantation of neurotrophic factor-transfected cells derived from Glia cell line on vascular dementia in rats and gene expression of Drebrin in hippocampal region.Methods By using gene clone technique,the GDNF gene was transfected into SH-SY5Y cell lines.104 adult male Sprague-Dawley rats weighing (200± 20) gram were divided into groups:transplanted group,injected group,control group,all of which accepted operation by permanent ligation of left common carotid artery and clipping right common carotid artery repeatedly to build up model of vascular dementia,and sham operation group which accepted no ligation or clipping.6 rats from each group were decapitated on the third day,seventh day and tenth day after transplanting treatment were for fluorescence detection.The rest 20 rats in each group were used to detect learning and memory functions by Morris water maze on the third day and decapitated on the fourth day after transplanting treatment.Then GDNF level in temporal lobe were detected by enzyme-linked immunosorbent assay (ELISA),while Drebrin mRNA and protein levels in hippocampal region were detected by real time-PCR and Westernblot respectively.Results There was strong fluorescent light detected around lateral ventricle of rats in transplanted group on the third day after transplantation,which faded on the seventh day and disappeared on the tenth day.The learning and memory functions of rats in transplanted group were improved significantly.The escape latency was shorter in transplanted group than in injected group and control group [(34.89±4.15) s vs.(43.86±6.95) s,(50.89±3.66) s,both P<0.05],while shuttle times through the third quadrant were more often in transplanted group than in injected group and control group [(11.00±1.49) vs.(9.26 ±1.38),(8.04 ± 1.12),both P<0.05].GDNF level and Drebrin mRNA and protein levels were higher in transplanted group than in injected group and control group [GDNF:(315.71±27.43) vs.(256.26±19.90),(141.95±21.33),Drebrin mRNA:(5.54±0.35) vs.(3.10±0.33),(1.32±0.23),Drebrinprotein:(0.55±0.05) vs.(0.43±0.06),(0.26±0.06),all P<0.05].Conclusions GDNF-transfected cells could survive in the lateral cerebral ventricle of rats for about seven days.The method for treating vascular dementia through the technique of transplanting GDNF-transfected cells is certain feasible,which has a better therapeutic effect than GDNF-injection directly into lateral cerebral ventricle.The therapeutic effect of GDNF on vascular dementia may be related to its action of regulating neural plasticity.
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Objective To observe the effectiveness of alprostedil in treatment of diabetic nephropathy and the mechanism of the renal protection of the drug. Methods One hundred patients with diabetic nephropathy due to diabetes mellitus were randomly divided into 2 groups ,50 in treatment group ,and 50 as controls, alprostadil was used intravenously in the Treatment Group for 12 day while routine management of diabetes such as food intake control,physical exercises and oral drugs or injection of insulin was conducted in both of the 2 groups. Urinary albumin were employed for evaluation as well as the side-effects. Results By the end of the trial, urinary albumin was decreased significantly compared with those before intravenous alprostadil treatment or with those in the control group (P<0.05). Conclusion Urinary albumin was decreased significantly after intravenous alprostadil. Therefore,it is a safe and effective drug in the treatment of diabetic nephropathy.
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Objective To investigate the feasibility of gene modification of peripheral blood derived endothelial progenitor cells(EPCs).Methods The rabbit mononuclear cells were collected from rabbit peripheral blood and cultured.The uptaking test of DiI-Ac-LDL was directly visualized with fluorescent microscopy,the immunofluorescence analysis was performed to detect the expression of surface marker.And the human tissue kallikrein(HTK)gene was amplified with PCR,and inserted into pEGFP-N3 vector.EPCs were transfected with the constructed plasmid by means of lipidosome,and the HTK-EGFP fused protein was visualized directly with fluorescent microscopy,and the expression of HTK was detected by RT-PCR and ELISA.Results After 5~6 days of culture,spindle-shaped attached cells clustered together,and cobblestone-like cell layer appeared after 15 days.Cells were positive of uptaking DiI-Ac-LDL and expressing vWF and CD133 related antigen.Plasmids were correctly formed and expressed in the rabbit EPCs.The fused protein had activity of both HTK and EGFP.Conclusion The HTK gene modified EPCs is potential in the treatment of artery injury and endothelial dysfunction.
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OBJECTIVE To evaluate the clinic feature and therapeutic principles and to improve curative effect of mucoepidemoid carcinoma originated in nasal cavity and nasal sinuses. METHODS A restrospective review of clinic features ,therapeutic methods and consequences was undertaken in thirty cases with nasal cavity and nasal sinuse mucoepidemoid carcinoma and treated at the 81st hospital of CPLA from 1990 to 2001. RESULTS Twenty patients received surgery、radiotherapy and chemotherapy, of which sixteen survived in five to ten years follow-up . Six patients received surgery and radiotherapy, of which four survived in five years follow-up. Only one out of two patients who received radiotherapy and chemotherapy respectively survived in five years follow-up. The total five years survival rate was 73 %(22/30). CONCLUSION CT or MRI scan may be helpful to evaluate the location and scale of mucoepidemoid carcinoma.Final diagnosis rely on histopathology examination. Early radical resection combined with radiotherapy and chemotherapy pre- or postoperatively could improve five-year-survival rate and reduce recurrence rate.
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OBJECTIVE To explore a more effective method of detecting sleep respiratory events in children. METHODS Thirty-eight children were tested with HypnoPTT and 31 with polysomnography (control). The test parameters and operative methods were compared. RESULTS In addition to the parameters comm.on to both HypnoPTT and polysomnography, inspiratory flow limitation and spontaneous microarousal can be measured using HypnoPTT and fewer electrodes were needed. CONCLUSION HypnoPTT is a convenient method characterized by less sleep disturbance and credible results, rendering it is especially suitable for pediatric cases. Pulse transit time is a useful parameter for diagnosing the sleep respiratory disease.
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Objective: To study the affection of Diallyl trisulfide on cultured human colon carcinoma cell line HT-29 and its mechanism. Methods: The viability of cells after various treatments were determined using the method of MTT. Flow cytometry were used to analyze the change of cell cycle and Bcl-2, Bax levels. Results: The proliferation of human colon carcinoma HT-29 cells were inhibited by Diallyl trisulfide. The inhibition was associated with dose and time dependence. Flow cytometry results showed that: 1) When cells were treated with Diallyl trisulfide at the concentration of 10 ?g/mL for 12 h and 24 h, the percentage of G0/G1 phase cells was decreased and that of G2/M phase cells was significantly increased compared with those in the control group.2) Diallyl trisulfide down regulated Bcl expression, while up-regulated bax expression. Conclusions: It was suggested that Diallyl trisulfide inhibited HT29 cell proliferation. And these were associated with arrest of cell cycles and down regulation of Bcl2/Bax ratio.
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AIM: To analyse the effect of Guipi Granule and serum that contains drug on rat's liver. METHODS: Isolated rat's liver perfusion model was used in the experiment for determining AST and ALT content. RESULTS: Middle and low concentration of Guipi Granule (2.5、5.0g/L) didn't cause any chang of enzyme AST、ALT content in isolated rat's liver perfusion liquid. The effect of serum was not different from Guipi Granule groups and control groups. CONCLUSION: The result shows that Guipi Granule doesn't cause damage to the liver cell.