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Abstract; Aim To solve the problems in the appliea- tion of egg yolk leeithin endotoxin test method, and and to establish the baeterial endotoxin examination method for egg yolk lecithin (for injeetion). Methods The ethanol solution of Tween 80 ( the volume ratio of tween 80 to anhydrous ethanol was 2. 5 • 2. 7, mixed for 4 min) was used to prepare lecithin solution of egg yolk at 0. 1 kg • L 1 , and 10 test water was added to 1 mL lecithin solution of egg yolk (500 EU • mL 1 standard solution of endotoxin IOjxL was added for pos¬itive control). After diluted 20 times with endotoxin test water, the standard curve range was 10 ~0. 01 EU • mL 1 by kinetic-turbidimetrie assay. Methodology of endotoxin test was studied using limulus lysate from two manufacturers and eight hatches of samples. Results The recoveries of eight hatches of samples all met the requirement of interference test between 50% and 200% stipulated in the pharmacopoeia, which solved the problems of the current endotoxin test method in practical application. Conclusions The bacterial en¬dotoxin test method of egg yolk lecithin with good dura-bility is established to provide the basis for the revision of pharmacopoeia.
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Aim To establish a batch of endotoxin standard for baeterial endotoxin detection of insoluble samples.Methods Candidate A and candidate B were prepared by freeze -drying bacterial endotoxin without excipient.The two batches of candidates were calibrated by three methods, including 13 laboratories for gel method, 9 laboratories for kinetic-turbidimetric assay and 5 laboratories for kinetic chromogenic assay.Results After statistical analysis, the geometric mean values of gel method, kinetic-turbidimetric assay and kinetic chromogenic assay calibration of candidate A were 680.1 EU, 827.0 EU and 800.8 EU, with RSD of 22.4%, 16.2% and 16.7%, respectively.The P value of variance analysis of calibration results of the three methods was 0.067, showing no significant difference.The weighted mean of potency was 774.0 EU (95% confidence interval 721.0 - 831.0, FL% 7.10).The geometric mean values of the calibration of candidate B by gel method, kinetic-turbidimetric assay and kinetic chromogenic assay method were 1 640.6 EU, 1 828.6 EU and 3 224.8 EU, with RSD of 33.9% , 47.0% and 54.4% , respectively.The P val¬ue of variance analysis of the calibration results of the three methods was 0.030, showing significant differ¬ence.Chi-square test was used to correct the weight of each method , and weighted average of the results of the three methods was used to obtain a corrected weighted average efficiency value of 1 822.7 EU (95% confi¬dence interval 1 548.7 -2 145.2, FL% 16.4).Can¬didate B was eliminated based on the results.Conclu¬sion Candidate A has become the first batch of na¬tional standard bacterial endotoxin (for insoluble sam¬ples only) approved by National Standard Substance Committee of China, and the potency is 700 EU.
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Objective: To evaluate the efficacy within the first 24 h post extracorporeal membrane pulmonary oxygenation (ECMO) and the impact of early efficacy on the prognosis of adult patients with fulminant myocarditis (FM). Methods: This retrospective case analysis study included hospitalized patients (age≥18 years) who were diagnosed with fulminant myocarditis from November 2016 to May 2021 in the First Affiliated Hospital of Zhengzhou University. Patients were divided into survival or non-survival groups according to treatment outcomes. The age, sex, treatments, drug use, ECMO use, clinical and laboratory data (before and 24 h after the use of ECMO) were analyzed. The change rate of clinical and laboratory data after 24 h use of ECMO was calculated to find differences between two groups. Multivariate logistic regression was used to analyze the related factors with in-hospital death and complication between the two groups. Results: A total of 38 FM patients treated with ECMO were included. There were 23 cases (60.5%) in the survival group, aged (39.6±13.7) years, and 17 (73.9%) cases were female. The total ECMO time was (134.4±71.3)h. There were 15 cases (39.5%) in non-survival group, aged (40.0±15.8) years, and there were 12(80.0%) female, the ECMO time was (120.1±72.4) h in this group. The proportion of tracheal intubation and continuous renal replacement therapy in the survivor group and dosage of norepinephrine within 24 h after ECMO implantation were significantly less than in non-survival group (all P<0.05). There was no significant difference in all efficacy related biochemical indexes between two groups before ECMO use. The levels of lactic acid, procalcitonin, creatinine, alanine aminotransferase, aspartate aminotransferase, creatine kinase-MB, cardiac troponin I and N-terminal B-type natriuretic peptide prosoma were significantly less in survival group than in non-survival group at 24 h after the use of ECMO (all P<0.05). Results of multivariate logistic regression analysis showed that the higher 24 h change rate of creatinine (OR=0.587, 95%CI 0.349-0.986, P=0.044) and creatine kinase-MB (OR=0.177, 95%CI 0.037-0.841, P=0.029) were positively correlated with reduced risk of in-hospital mortality. The central hemorrhage and acute kidney injury in survival group were less than in non-survivor group (P<0.05). Conclusions: After 24 h early use of ECMO in FM patients, the improvement of various efficacy related biochemical test indexes in the survival group was better than that in the non-survival group. Faster reduction of creatine kinase-MB and creatinine values within 24 h ECMO use is positively correlated with reduced risk of in-hospital mortality in adult patients with FM.
Subject(s)
Adolescent , Adult , Female , Humans , Middle Aged , Young Adult , Extracorporeal Membrane Oxygenation/methods , Hospital Mortality , Myocarditis/therapy , Retrospective Studies , Treatment OutcomeABSTRACT
Objective: To investigate the related factors of left ventricular thrombus (LVT) formation within two weeks in patients with acute ST-segment elevation myocardial infarction (STEMI) and left ventricular aneurysm (LVA). Methods: Consecutive inpatients with acute STEMI and left ventricular aneurysm, hospitalized from January 2014 to June 2020 in the First Affiliated Hospital of Zhengzhou University, were enrolled in this retrospective study. Patients were divided into LVT group and non-LVT group according to the presence or absence of LVT. The clinical data, echocardiography findings, coronary angiography and treatments were compared between the two groups. Subgroup analysis was performed on the patients receiving primary percutaneous coronary intervention (PCI). Onset to door, door to balloon, onset to balloon time were also compared. Multivariate logistic regression was used to analyze the related factors of LVT formation early post STEMI in enrolled patients. The effects of each index in multivariate logistic regression analysis were illustrated in the forest plot. Results: A total of 144 patients were included, there were 52 cases (36.1%) in LVT group, the age was (56.4±11.2) years, and 46 (88.5%) cases were male. There were 92 cases (63.9%) in non-LVT group, the age was (61.7±11.5) years, and there were 73 (79.3%) males in this group. The patients in LVT group were younger than those in non-LVT group, the proportion of anemia, history of heavy drinking, history of chronic kidney disease, left ventricular ejection fraction (LVEF) ≤40%, preoperative thrombosis in myocardial infarction (TIMI) blood flow grade 0 and postoperative TIMI blood flow grade ≤2 were significantly higher in LVT group than those in non-LVT group (all P<0.05). The incidence of history of angina, collateral circulation of the coronary arteries, primary PCI or venous thrombolysis were significantly lower in LVT group than those in non-LVT group (all P<0.05). The percentage of anti-platelet therapy and anticoagulant therapy was similar between the two groups (all P˃0.05). Subgroup analysis in patients undergoing primary PCI showed that onset to door, door to balloon, onset to balloon time were similar between the two groups. Multivariate logistic regression analysis and forest plot showed that history of heavy drinking (OR=6.982, 95%CI 1.501-32.469, P=0.013), anemia (OR=3.373, 95%CI 1.075-10.585, P=0.037), LVEF≤40% (OR=3.016, 95%CI 1.027-8.859, P=0.045), preoperative TIMI blood flow grade 0 (OR=3.311, 95%CI 1.214-9.029, P=0.019) were positively correlated with LVT in patients with acute STEMI and LVA. History of angina (OR=0.159, 95%CI 0.058-0.441, P<0.001), collateral circulation of the coronary arteries (OR=0.189, 95%CI 0.053-0.673, P=0.010), primary PCI or venous thrombolysis (OR=0.252, 95%CI 0.093-0.682, P=0.007) were negatively correlated with LVT in patients with acute STEMI and LVA. Conclusions: History of heavy drinking, anemia, LVEF ≤40%, preoperative TIMI blood flow grade 0 are associated with increased risk of the LVT in patients with acute STEMI and LVA at early stage of the disease. However, history of angina, collateral circulation of the coronary arteries, primary PCI or venous thrombolysis are associated with lower risk of the LVT in these patients.
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Myocardial Infarction , Percutaneous Coronary Intervention , Retrospective Studies , ST Elevation Myocardial Infarction , Stroke Volume , Thrombosis , Ventricular Function, LeftABSTRACT
Finding the novel drug from the effective components of traditional Chinese herbal medicine is a hotspot of the modern pharmacological research.Hyperoside (HYP) belongs to flavonoid glycosides,and it has various properties,such as anti-inflammation,anti-spasm,anti-diuretic,antitussive,lowering blood pressure,and lowering cholesterol effects as well as protective effects for the cardiac and cerebral blood vessels.The purpose of this study was to investigate the effects of HYP on inflammatory and apoptotic responses in vascular endothelial cells stimulated by lipopolysaccharide (LPS) and further to identify the possible mechanisms underlying these effects.In our study,human umbilical vein endothelial cells (HUVECs) were stimulated with 1 μg/mL LPS in the presence or absence of HYP (10,20 and 50 μrnol/L).Our results indicated that HYP alone exerted no cytotoxicity on HUVECs,while it had an up-regulatory effect on the viability of HUVECs induced by LPS in a dose-dependent manner;increased mRNA expression of IL-1β,IL-6,TNFα and iNOS induced by LPS was attenuated after treatment with HYP both in a dose-and time-dependent manner;LPS-induced HUVECs apoptosis and cleaved-caspase 8,9,3 were all significantly reduced by HYP.Furthermore,the possible pathway involved in apoptosis and inflammation by HYP was detected,and the results showed that when treated with HYP,LPS-induced mitochondrial membrane instability was significantly inhibited through up-regulation of Bcl-2 and down-regulation of Bax.Furthermore,the expression of TLR4 and the phosphorylation of Iκ Bα and p65 in LPS-treated cells were blocked by HYP.Our results suggested that HYP treatment prevented HUVECs from LPS-induced inflammation and apoptosis responses,which might be mediated by inhibiting TLR4/NFκB pathway.
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Finding the novel drug from the effective components of traditional Chinese herbal medicine is a hotspot of the modern pharmacological research.Hyperoside (HYP) belongs to flavonoid glycosides,and it has various properties,such as anti-inflammation,anti-spasm,anti-diuretic,antitussive,lowering blood pressure,and lowering cholesterol effects as well as protective effects for the cardiac and cerebral blood vessels.The purpose of this study was to investigate the effects of HYP on inflammatory and apoptotic responses in vascular endothelial cells stimulated by lipopolysaccharide (LPS) and further to identify the possible mechanisms underlying these effects.In our study,human umbilical vein endothelial cells (HUVECs) were stimulated with 1 μg/mL LPS in the presence or absence of HYP (10,20 and 50 μrnol/L).Our results indicated that HYP alone exerted no cytotoxicity on HUVECs,while it had an up-regulatory effect on the viability of HUVECs induced by LPS in a dose-dependent manner;increased mRNA expression of IL-1β,IL-6,TNFα and iNOS induced by LPS was attenuated after treatment with HYP both in a dose-and time-dependent manner;LPS-induced HUVECs apoptosis and cleaved-caspase 8,9,3 were all significantly reduced by HYP.Furthermore,the possible pathway involved in apoptosis and inflammation by HYP was detected,and the results showed that when treated with HYP,LPS-induced mitochondrial membrane instability was significantly inhibited through up-regulation of Bcl-2 and down-regulation of Bax.Furthermore,the expression of TLR4 and the phosphorylation of Iκ Bα and p65 in LPS-treated cells were blocked by HYP.Our results suggested that HYP treatment prevented HUVECs from LPS-induced inflammation and apoptosis responses,which might be mediated by inhibiting TLR4/NFκB pathway.
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The influence on 10 kinds of ginsensides of different processed methods of Panacis Quinquefolii Radix was discussed. White Panacis Quinquefolii Radix (sliced and dried at -80 °C), red Panacis Quinquefolii Radix( steamed, sliced and dried at -80 °C) and commercial Radix Panacis Quinquefolii (dried by electric blast air) processed by different methods. HPLC-PDA-ESI- MS method was established before by our team. Ten kinds of ginsenosides of them were determined. The content of total ginsenosides were as follow: commercial Panacis Quinquefolii Radix > white Panacis Quinquefolii Radix > red Panacis Quinquefolii Radix. Compared with white Panacis Quinquefolii Radix, the content of Re, Rc, Rb3 and Rb2 of Red Radix Panacis Quinquefolii decreased but increased that of Rg,, Rb1. Both Rg2 and Rg, were not found in white Panacis Quinquefolii Radix and commercial Panacis Quinquefolii Radix by PDA detector, and low response in ESI-MS, while red Panacis Quinquefolii Radix was to the high content that of 0. 027% and 0.040 1%. The constituent of RA0 of red Panacis Quinquefolii Radix was higher than the other two. After Panacis Quinquefolii Radix processed, the kind and content of ginsensides were significantly changed. The constituent of some kinds of ginsensides was increased and some decreased. Rf was not found in all Panacis Quinquefolii Radix samples which were consistent with the former documents.
Subject(s)
Chemistry, Pharmaceutical , Methods , Ginsenosides , Chemistry , Mass Spectrometry , Panax , Chemistry , Plant Extracts , Chemistry , Plant Roots , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To observe the efficacy of integrative medical sequential method in treating cerebral palsy (CP) children's intelligence development, muscular tension, serum interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-alpha).</p><p><b>METHODS</b>Totally 111 CP children were randomly assigned to the control group (50 cases) and the treatment group (61 cases). All patients received comprehensive rehabilitation training and intravenous dripping of Monosialotetrahexosylganglioside Sodium Injection for 10 days. But those in the treatment group additionally received Chinese medical enema for brain resuscitation, relieving rigidity of muscles and activating collaterals for 14 days. Then they started another medication cycle and lasted for a total of 6 cycles. Serum IL-6 levels and TNF-alpha contents were determined before treatment. Scoring for muscular tension, Gesell score for intelligence development, contents of serum IL-6 and TNF-alpha were assessed before and after treatment in the two groups.</p><p><b>RESULTS</b>Compared with before treatment in this group, muscular tension, Gesell scores for intelligence development all decreased in the two groups (P < 0.05). As for inter-group comparison, the decrement was more obvious in the treatment group than in the control group (P < 0.05). The total effective rate was 86.9% in the treatment group and 76.0% in the control group (P < 0.05). The contents of IL-6 and TNF-alpha were obviously reduced in the treatment group and the control group after treatment (P < 0.01). The decrement was more obvious in the treatment group (P < 0.05).</p><p><b>CONCLUSION</b>The two treatment methods were effective for CP children, but the efficacy was superior in the treatment group than in the control group, indicating integrative medical methods could play a synergistic effect and optimize the treatment program for CP.</p>
Subject(s)
Child, Preschool , Female , Humans , Infant , Male , Cerebral Palsy , Drug Therapy , Drugs, Chinese Herbal , Therapeutic Uses , Gangliosides , Therapeutic Uses , Integrative Medicine , Intelligence , Interleukin-6 , Blood , Phytotherapy , Tumor Necrosis Factor-alpha , BloodABSTRACT
BACKGROUND/AIMS: Upregulated CD64 expression on neutrophils is the most useful marker for acute bacterial infections and systemic inflammation. However, it is unknown whether CD64 is involved in the pathogenesis of acute pancreatitis (AP). This study was designed to determine whether CD64 is implicated in severe acute pancreatitis (SAP), and thus, is a suitable marker for SAP. METHODS: SAP was induced in rats with an intraperitoneal injection of L-arginine. CD64 expression in the rat pancreas was determined by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry. Additionally, the CD64 mRNA expression in peripheral blood leukocytes from 21 patients with mild acute pancreatitis (MAP) and 10 patients with SAP was investigated at the time of admission and during remission by qRT-PCR. RESULTS: CD64 mRNA and protein expression in the pancreas was significantly higher in rats with SAP, compared to the controls. The CD64 expression was higher in the patients with SAP than in the patients with MAP. During remission, CD64 mRNA decreased in both the MAP and SAP patients. The area under the curve of CD64 expression for the detection of SAP was superior to both the Ranson and the Acute Physiology and Chronic Health Evaluation II scores. CONCLUSIONS: The CD64 level was significantly increased in correlation with the disease severity in SAP and may act as a useful marker for predicting the development of SAP.
Subject(s)
Adolescent , Adult , Aged , Animals , Female , Humans , Male , Middle Aged , Young Adult , Acute Disease , Arginine/toxicity , History, Ancient , Immunohistochemistry , Pancreatitis/metabolism , RNA, Messenger/metabolism , Rats, Sprague-Dawley , Receptors, IgG/metabolism , Up-RegulationABSTRACT
BACKGROUND/AIMS: To investigate the differential expression of RING finger (RNF) proteins in Barrett esophagus (BE) and esophageal adenocarcinoma (EAC). METHODS: The differential expression of RNFs in normal esophagus (NE), BE, and EAC was screened using microarray assay. Real-time quantitative polymerase chain reaction (PCR), tissue microarray assay, and Western blot analysis were independently performed to detect the mRNA and protein expression of screened RNFs. RESULTS: The expression of nine RNFs in the BE or EAC was 2-fold higher than those in NE. Among these proteins, the RNF32 and RNF121 expression in BE was 20.3-fold and 16.4-fold higher, respectively, than that in NE, and the expression of RNF24, RNF130, RNF141, RNF139, RNF11, RNF14, and RNF159 was upregulated more than 2-fold compared with NE. The expression of nine RNFs was not only upregulated in the EAC but was also positively related to the RNF expression in BE. The PCR results also indicated increased expression of these RNFs in BE and EAC compared to NE. Furthermore, the mRNA expression of all RNFs, except for RNF141 in EAC, was dramatically higher than those in the BE. Similar results were also obtained from the Western blot analysis. CONCLUSIONS: A total of nine RNFs play critical roles in the progression of BE to EAC.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Adenocarcinoma/enzymology , Barrett Esophagus/enzymology , Carrier Proteins/genetics , Disease Progression , Esophageal Neoplasms/enzymology , Gene Expression Profiling , Intracellular Signaling Peptides and Proteins/genetics , Proteins/genetics , RING Finger Domains , Receptors, Cell Surface/genetics , Ubiquitin-Protein Ligases/geneticsABSTRACT
<p><b>BACKGROUND</b>Everolimus, a derivative of sirolimus, is a potent immunosuppressant that has important anti-proliferative properties. In the present study, we demonstrated the inhibiting neointimal hyperplasia in injured carotid arteries in rats by using two different doses of everolimus administrated via the oral route for a long time.</p><p><b>METHODS</b>A rat model of carotid artery injury was established by balloon inflation. Eighty rats were randomly divided into the sham-operated group (n = 20), injury group (n = 20), low dosage of everolimus group (n = 20), and high dosage of everolimus group (n = 20). The low dose of everolimus (1.5 mg/kg) was given one day before injuring the carotid artery by balloon, followed by 0.75 mg/kg per day for 28 days via intragastric gavage. High dose everolimus (2.5 mg/kg) was given one day before injuring the carotid artery by balloon, followed by 1 mg/kg per day for 28 days. Expression of eukaryotic translation initiation factor 4E (eIF-4E) and phosphorylation of ribosomal protein S6 kinase 1 (P70S6K) were determined by reverse transcription-polymerase chain reaction and Western blotting analysis.</p><p><b>RESULTS</b>In the injured carotid artery, neointimal hyperplasia was normally observed four weeks after injury. Everolimus inhibited neointimal hyperplasia after balloon injury in a dose dependent manner. At the same time, the study demonstrated that everolimus reduced the expression of P-P70S6K, eIF-4E, transforming growth factor (TGF)-β1 and of proliferating cell nuclear antigen (PCNA).</p><p><b>CONCLUSIONS</b>Everolimus significantly inhibited neointimal hyperplasia of the injured carotid artery. The effect depended on dosage and was associated with the reduction of phosphorylation of P70S6K and the eIF-4E expression level.</p>
Subject(s)
Animals , Male , Rats , Carotid Arteries , Carotid Artery Injuries , Drug Therapy , Carrier Proteins , Metabolism , Everolimus , Neointima , Drug Therapy , Phosphoproteins , Metabolism , Phosphorylation , Rats, Sprague-Dawley , Ribosomal Protein S6 Kinases, 70-kDa , Metabolism , Sirolimus , Therapeutic UsesABSTRACT
Small ubiquitin-related modifiers (SUMOs) belong to an important class of ubiquitin like proteins. SUMOylation is a post-translational modification process that regulates the functional properties of many proteins, among which are several proteins implicated in neurodegenerative diseases. This study was aimed to investigate the changes of SUMO-1 expression and modification, and the relationship between SUMO-1 and Alzheimer's disease (AD) pathology in APP/PS1 transgenic AD mice. Using Western blot, co-immunoprecipitation and immunofluorescent staining methods, the SUMO-1 expression and modification and its relation to tau, amyloid precursor protein (APP) and β-amyloid protein (Aβ) in the 12-month-old APP/PS1 transgenic AD mice were analyzed. The results showed that: (1) Compared with the normal wild-type mice, the expression and modification of SUMO-1 increased in brain of AD mice, which was accompanied by an increase of ubiquitination; (2) In RIPA soluble protein fraction of cerebral cortex, co-immunoprecipitation analysis showed tau SUMOylated by SUMO-1 increased in AD mice, however, AT8 antibody labeled phosphorylated tau was less SUMOylated whereas PS422 antibody labeled phosphorylated tau was similar to control mice; (3) Double immunofluorescent staining showed that SUMO-1 could distributed in amyloid plaques, appearing that some of SUMO-1 diffused in centre of some plaques and some of SUMO-1 co-localized with AT8 labeled phosphorylated tau forming punctate aggregates around amyloid plaques which was concerned as dystrophic neurites, however, less Aβ, APP and PS422 labeled phosphorylated tau were found co-localized with SUMO-1. These results suggest that SUMO-1 expression and modification increase abnormally in transgenic AD mice, which may participate in modulation of the formation of senile plaques and dystrophic neurites.
Subject(s)
Animals , Mice , Alzheimer Disease , Amyloid beta-Peptides , Metabolism , Amyloid beta-Protein Precursor , Metabolism , Brain , Pathology , Mice, Transgenic , Neurites , Pathology , Phosphorylation , Plaque, Amyloid , SUMO-1 Protein , Metabolism , Sumoylation , tau Proteins , MetabolismABSTRACT
Objective To compare 2 kinds of low-dose in adrenocorticotropic hormone(ACTH) stimulation tests for evaluating adrenal function in children with primary nephrotic syndrome(PNS).Methods Seventeen patients (dose of prednisone < 1 mg/kg) with PNS (PNS group) and 7 control subjects (control group) were enrolled.On the first day,all subjects were injected ACTH intravenously at 8 ∶ 00.Serum cortisol was measured just before the ACTH administration,after 20,30 minutes(the first method).On the second day,all subjects were injected ACTH intravenously at 14 ∶ 00.After the ACTH injection,blood was taken every 10 minutes for a period of 1 h to measure serum cortisol (the second method).Adrenal function was evaluated.Cortisol levels of 497 nmol/L or more were taken as normal adrenal response to low dose of ACTH stimulation.Results In the control group,all subjects showed normal adrenal function with the first method and the second method.Cortisol levels at 30 minutes were all > 497 nmol/L.Cortisol levels at 0,20 and 30 min points with the first method did not differ from those with the second method(all P > 0.05).In 17 PNS patients,3 cases (17.65%)and 7 cases (41.18%)showed subnormal responses with the first method and the second method,respectively.Detection rate of subnormal responses was significantly higher with the second method (P <0.05).In the first method,5 cases of the 17 patients showed subnormal responses at 20 minutes and 2 of above 5 cases turned to normal responses at 30 minutes.However,10 cases of the 17 patients showed subnormal responses at 20 minutes and none of above 10 cases achieved normal responses at 30 minutes with the second method.Comparison of function of adrenal cortex at 20 and 30 minutes showed that there were difference in the first and second method(P =0.025,0.008,respectively).Comparison of cortisol levels at 0,20 and 30 minutes in the first and the second method showed that there were difference at 0 minute and amplification of 20-30 minutes and there was no difference at any other points.Conclusions The low dose of ACTH test in the second method for evaluation of adrenocortical function in children with PNS is more accurate and sensitive.
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<p><b>OBJECTIVE</b>To investigate the modulation effects of mesenchymal stem cells (MSC) implantation on the myofibroblasts congregating in the infarct region after myocardial infarction (MI).</p><p><b>METHODS</b>MI was induced in SD rats by left anterior descending coronary artery ligation, and the experimental animals were assigned randomly into the sham group, MI + PBS group and MI + MSC group (myocardial injection of 0.1 ml 2×10(7)/ml in four locations in the infarct region). Echocardiography, hemodynamic examinations and Masson trichrome staining were performed. Implanted MSC differentiation and myofibroblasts congregating in infarct region were investigated by immunofluorescence staining. TGF-β(1)-Smad2 signaling pathway was examined by real-time RT-PCR and Western blot.</p><p><b>RESULTS</b>(1) Four weeks late, heart-weight/body-weight ratio [(3.04 ± 0.16) mg/g vs. (3.34 ± 0.14) mg/g, P < 0.01] and myocardial infarction size [(38.72 ± 2.38)% vs. (46.36 ± 2.81)%, P < 0.01] were significantly reduced in MI + MSC group than in MI + PBS group, while scar thickness of infarct region was thicker [(0.93 ± 0.17) mm vs. (0.65 ± 0.16) mm, P = 0.01], and LVEF was higher [LVEF: (32.5 ± 5.9)% vs. (26.5 ± 4.5)%, P = 0.03] in MI + MSC group than in MI + PBS group. (2) Myofibroblasts congregating in the infarct region was significantly enhanced in MI + MSC group compared with MI + PBS group [(196 ± 20) cells/mm(2) vs. (89 ± 25) cells/mm(2), P < 0.01], and part of implanted MSC expressed α-SMA(+). (3) TGF-β(1) expression and the phosphorylating of Smad2 in the infarct region were significantly upregulated in MI + MSC group compared with MI + PBS group (all P < 0.05).</p><p><b>CONCLUSIONS</b>MSC could improve myocardial function and promote myofibroblasts congregating in the infarct region via activating the TGF-β(1)-Smad2 signaling pathway in this model.</p>
Subject(s)
Animals , Male , Rats , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Myocardial Infarction , Metabolism , Therapeutics , Myofibroblasts , Cell Biology , Metabolism , Rats, Sprague-Dawley , Transforming Growth Factor beta1 , Metabolism , Ventricular RemodelingABSTRACT
<p><b>BACKGROUND</b>Metformin has become a cornerstone in the treatment of patients with type-2 diabetes. Accumulated evidence suggests that metformin supports direct cardiovascular effects. The present study aimed to investigate if metformin has beneficial effects on primary cardiomyocytes damaged by H2O2, and reveal the potential mechanism of action of metformin.</p><p><b>METHODS</b>Cardiomyocytes were incubated in the presence of 100 µmol/L H2O2 for 12 hours. Cardiomyocytes were pretreated with metformin at different concentrations and time and with aminoimidazole carboxamide ribonucleotide (AICAR) (500 µmol/L), an adenosine monophophate (AMP)-activated protein kinase (AMPK) agonist for 60 minutes before the addition of H2O2. Other cells were preincubated with compound C (an AMPK antagonist, 20 µmol/L) for 4 hours. The viability and apoptosis of cells were analyzed. AMPK, endothelial nitric oxide synthase (eNOS), and transforming growth factor (TGF)-β1 were analyzed using immunblotting.</p><p><b>RESULTS</b>Metformin had antagonistic effects on the influences of H2O2 on cell viability and attenuated oxidative stress-induced apoptosis. Metformin also increased phosphorylation of AMPK and eNOS, and reduced the expression of TGF-β1, basic fibroblast growth factor (bFGF), and tumor necrosis factor (TNF)-α.</p><p><b>CONCLUSIONS</b>Metformin has beneficial effects on cardiomyocytes, and this effect involves activation of the AMPK-eNOS pathway. Metformin may be potentially beneficial for the treatment of heart disease.</p>
Subject(s)
Animals , Rats , AMP-Activated Protein Kinases , Physiology , Aminoimidazole Carboxamide , Pharmacology , Apoptosis , Cell Survival , Cells, Cultured , Hypoglycemic Agents , Pharmacology , Metformin , Pharmacology , Myocytes, Cardiac , Metabolism , Nitric Oxide Synthase Type III , Genetics , RNA, Messenger , Rats, Wistar , Ribonucleotides , Pharmacology , Transforming Growth Factor beta1 , Genetics , Tumor Necrosis Factor-alpha , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of acupuncture on the treatment of glaucoma so as to find out a kind of simple and effect approach to control and stabilize intraocular pressure (IOP) of patients with glaucoma.</p><p><b>METHODS</b>Thirty-nine cases (75 eyes) of glaucoma (unstable control of IOP) were treated with acupuncture at Sibai (ST 2), Cuanzhu (BL 2), Jingming (BL 1), Qiuhou (EX-HN 7), etc. IOP before and after acupuncture, at 10 a.m, 2 p.m, 6 p.m, 10 p.m., at 5 a.m. and 7 a.m. on the second day as well as diurnal IOP variation in 24 h were observed separately.</p><p><b>RESULTS</b>After acupuncture, IOP at each time point reduced as compared with that before acupuncture. IOP at 5 a.m., 7 a.m., 10 a.m., 6 p.m. and 10 p.m. after acupuncture was different significantly in statistics as compared with that before acupuncture correspondingly (all P<0.05). After acupuncture, 24 h diurnal IOP variation was (5.31 +/- 2.84) mmHg, which was lower significantly than (7.06 +/- 3.86) mmHg before acupuncture (P<0.05).</p><p><b>CONCLUSION</b>For the patients with unstable control of IOP, acupuncture can not only reduce IOP, but also stabilize 24 h diurnal IOP variation and benefit the visual function of patients with glaucoma.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Acupuncture Therapy , Glaucoma , Therapeutics , Intraocular PressureABSTRACT
Arginine vasopressin (AVP) has been implicated in the pathophysiology of cardiac hypertrophy. We previously demonstrated that AVP is a mitogen for neonatal rat cardiac fibroblasts (CFs). In the present study, we extend our investigation to adult rat CFs to explore whether AVP could induce adult rat CFs proliferation and, if so, to identify the underlying mechanisms. We found that AVP stimulated cell proliferation, an effect abolished by V1 receptor antagonist, d(CH2)5[Tyr2(Me), Arg8]-vasopressin, but not V2 receptor antagonist, desglycinamide-[d(CH2)5, D-Ile2, Ile4, Arg8]-vasopressin. AVP also activated extracellular signal-regulated kinase 1/2 (erk1/2), a response mimicked by protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA), but abolished by depleting cellular PKC through chronic PMA incubation. Calphostin C, an inhibitor of PKC, attenuated and PMA mimicked the effect of AVP on cell proliferation, whereas Ca2+ chelating agent 1,2-bis(2-aminophenoxy)ethane N, N, N', N'-tetraacetic acid (BAPTA) had no effect. Further, AVP downregulated protein expression of p27Kip1, increased cyclins D1, A, and E expressions and induced cell cycle progression through G0/G1 into S stage. Antisense oligonucleotides against cyclins D1, A, and E decreased cell number in the presence of AVP. Whereas antisense treatment against p27Kip1 and overexpression of p27Kip1 exerted a stimulatory and inhibitory effect, respectively. Inhibiting erk1/2 activation by PD98059 abolished the effect of AVP on cell proliferation, cell cycle regulatory proteins, and cell cycle progression. These results suggest that AVP is a mitogen for adult rat CFs via the mediation of V1 receptor and PKC-erk1/2 pathway.
Subject(s)
Arginine Vasopressin/physiology , Fibroblasts/cytology , Mitogen-Activated Protein Kinase 1/physiology , Mitogen-Activated Protein Kinase 3/physiology , Myocardium/cytology , Protein Kinase C/physiology , Animals , Antidiuretic Hormone Receptor Antagonists , Arginine Vasopressin/pharmacology , Cell Proliferation , Cells, Cultured , Fibroblasts/drug effects , Fibroblasts/metabolism , Male , Mitosis , Rats , Rats, Sprague-Dawley , Receptors, Vasopressin/metabolismABSTRACT
<p><b>OBJECTIVE</b>To analyze the baseline data of outpatient clinical subjects with vertigo and study on the clinical characteristics of vertigo.</p><p><b>METHOD</b>The questionnaires and clinical tests data of 3432 patients complained vertigo were retrospectively analyzed.</p><p><b>RESULTS</b>All the patients received interview and vestibular function test. These patients aged 4-89 years with an average age of (40 +/- 18.6) years. Among them 1513 (44.09%) were male and 1919 (55.91%) were female, with a male:female ratio of 1:1.27. Vertigo patients increased according to age and reached its peak in the 41-60 years among all patients. The incidence might increase along with the increase of education level in urban populations. The onset of vertigo might correlate with the careers but differed among different populations.</p><p><b>CONCLUSIONS</b>Vertigo attacks patients in all age spans, but vertigo is highly prevalent in the population aged 41-60 years. The onset of vertigo is related to many different factors.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Age Distribution , Retrospective Studies , Surveys and Questionnaires , Vertigo , Epidemiology , Vestibular Function TestsABSTRACT
Arginine vasopressin (AVP), a neurohormone and hemodynamic factor implicated in the pathophysiology of hypertension and congestive heart failure, can also act as a growth-stimulating factor. Our previous work demonstrated that AVP is a mitogen for neonatal rat cardiac fibroblasts (CFs). In the present study, we extended our investigations to adult rat CFs to explore whether AVP could induce adult rat CF proliferation and, if so, to identify the mechanism involved. Adult rat CFs were isolated, cultured and subjected to AVP treatment. DNA synthesis and cell cycle distribution were analyzed by [(3)H]-thymidine incorporation and flow cytometry. Cellular extracellular signal-regulated kinase 1/2 (ERK1/2) activity was measured by in vitro kinase assay using myelin basic protein (MBP) as a substrate. Protein expressions of total- and phospho-ERK1/2, p27(Kip1), cyclins D1, A, E were assessed by Western blot. The results showed that AVP stimulated DNA synthesis in adult rat CFs, and the effect was abolished by a V1 receptor antagonist, d(CH(2))(5)[Tyr(2)(Me), Arg(8)]-vasopressin (0.1 μmol/L), but not by a V2 receptor antagonist, desglycinamide-[d(CH(2))(5), D-Ile(2), Ile(4), Arg8]-vasopressin (0.1 μmol/L). AVP induced an activation of ERK1/2, which could be mimicked by the protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA, 30 nmol/L, 5 min), but abolished by depletion of PKC via chronic PMA incubation (2.5 μmol/L, 24 h). In addition, AVP down-regulated protein expression of p27(Kip1), increased protein expressions of cyclins D1, A and E, and induced cell cycle progression from G(0)/G(1) into S stage. Inhibition of ERK1/2 activation by PD98059 (30 μmol/L) abolished the effect of AVP on DNA synthesis, protein expressions of p27(Kip1), cyclins D1, A and E as well as cell cycle progression. These results suggest that AVP is also a growth factor for adult rat CFs. The mitogenic effect of AVP is mediated via V1 receptors and PKC-ERK1/2 pathway. Moreover, AVP modulates the expressions of cell cycle regulatory proteins p27(Kip1) and cyclins D1, A and E, which lie downstream of ERK1/2 activation, and induces cell cycle progression in adult rat CFs.
Subject(s)
Animals , Rats , Antidiuretic Hormone Receptor Antagonists , Pharmacology , Arginine Vasopressin , Pharmacology , Cell Cycle , Cell Cycle Proteins , Metabolism , Cell Proliferation , Fibroblasts , Cell Biology , Mitogen-Activated Protein Kinase 3 , Metabolism , Myocardium , Cell Biology , Phosphorylation , Protein Kinase C , Metabolism , Signal Transduction , Tetradecanoylphorbol Acetate , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To design an intelligent speech test system with reliability and convenience using the computer software and to evaluate this system.</p><p><b>METHODS</b>First, the intelligent system was designed by the Delphi program language. Second, the seven monosyllabic word lists recorded on CD were separated by Cool Edit Pro v2.1 software and put into the system as test materials. Finally, the intelligent system was used to evaluate the equivalence of difficulty between seven lists. Fifty-five college students with normal hearing participated in the study.</p><p><b>RESULTS</b>The seven monosyllabic word lists had equivalent difficulty (F = 1.582, P > 0.05) to the subjects between each other and the system was proved as reliability and convenience.</p><p><b>CONCLUSIONS</b>The intelligent system has the feasibility in the clinical practice.</p>