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Since December 2019, coronavirus disease 2019 (COVID-19) has spread rapidly in China. National Health Commission of the People’s Republic of China and local departments have released a number of diagnosis and treatment plans for COVID-19. Some plans recommend Xiyanping Injection for the clinical treatment of COVID-19. The active component of the drug is the total sulfonate of andrographolide, which is a derivative of andrographolide. This paper summarized the pharmacological action and clinical application of andrographolide, and proposed that it has the functions of clearing heat and detoxification, anti-bacterial and anti-inflammatory, and definite therapeutic effect on respiratory system diseases including viral pneumonia and upper respiratory tract infection, and a large number of clinical data have been accumulated. Andrographolide has a potential antiviral effect on the treatment of COVID-19, and can reduce the level of inflammation in patients, improve respiratory symptoms, inhibit concurrent bacterial infection, and improve the body immunity. At the same time, it will not bring the immunosuppressive effect of hormone drugs, and the incidence of adverse reactions is low. In addition, Andrographolide has certain hepatoprotective effects and clinical value for treating cardiovascular diseases in clinical experience, suggesting that it may have some protective effects on drug-induced liver injury, heart injury and liver injury caused by durgs against COVID-19, but further clinical verification is needed. The pharmacological action and clinical application of andrographolide are summarized. This paper also analyzes the etiology, pathogenesis and dialectical treatment of COVID-19 from the perspective of traditional Chinese medicine, and points out that the treatment of COVID-19 with andrographolide is consistent with the theory of traditional Chinese medicine.
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Objective@#To explore the prognostic effects of mean corpuscular volume (MCV) in patients with myelodysplastic syndromes (MDS) .@*Methods@#321 newly diagnosed, untransfused primary MDS patients who administered from December 2009 to December 2017 were enrolled. The association of MCV with prognosis and several clinical features and genetic mutations were analyzed.@*Results@#Patients were divided into MCV≤100 fl (n=148) and MCV>100 fl (n=173) cohorts. Median overall survival of patients with MCV≤100 fl was shorter than their counterparts (27 months vs 72 months, P<0.001) . In subgroup analysis, MCV≤100 fl patients had worse survivals in bone marrow blast <5% cohort (34 months vs not reached, P=0.002) , but not so in ≥5 % cohort (17 months vs 20 months, P=0.078) . MCV≤100 fl was still an independent adverse variable (HR=1.890, 95%CI 1.007-3.548, P=0.048) after adjusting for clinical and laboratory variables and mutation topography in bone marrow blasts<5% cohort. In bone marrow blasts<5% cohort, patients with MCV≤100 fl had higher hemoglobin levels [90 (42-153) g/L vs 78.5 (28-146) g/L, P=0.015].The proportions of Revised International Prognostic Scoring System (IPSS-R) high/very high risks and poor/very poor IPSS-R karyotypes were higher in MCV≤100 fl cohort (28.8% vs 10.8%, P=0.003; 24.7% vs 12.9%, P=0.049) . MCV≤100 fl cohort had more genetic mutations than those with MCV>100 fl though without significance (0.988 vs 0.769, P=0.064) . Mutated SF3B1 was less frequently in MCV≤100 fl cohort (4.7% vs 15.4%, P=0.018) .@*Conclusion@#MCV≤100 fl was an independent adverse variable after adjusting for clinical and laboratory variables and mutation topography in MDS patients with bone marrow blasts<5%.
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Objective@#To explore the effects and molecular mechanism of the selective JAK1inhibitor SHR0302 and Ruxolitinib on myeloproliterative neoplasms (MPN) cell line SET2 and primary cells in vitro.@*Methods@#Cell proliferation was detected by CCK8 kit. Colony forming experiment was conducted to evaluate erythroid burst colony formation unit (BFU-E) of primary cells from MPN patients. Multi-factor kits were used to detect six inflammatory cytokines. Phosphorylated proteins of Jak-Stat signaling pathway were tested by Western blot.@*Results@#At different time points after treated with SHR0302 and Ruxolitinib, the inhibition of cell proliferation was dose dependent by both drugs (P<0.01) . The inhibitory rates of 2.5 μmol/L SHR0302 and 0.1 μmol/L Ruxolitinib on SET2 cells for 72 h were comparable, i.e. (59.94±0.60) % and (64.00±0.66) %, respectively, suggesting that the inhibitory effect of SHR0302 was weaker than that of Ruxolitinib. Similarly, both SHR0302 and Ruxolitinib inhibited BFU-E in primary marrow cells from MPN patients in a dose-dependent manner. SHR0302 1.0 μmol/L produced similar degree of inhibition compared to Ruxolitinib 0.2 μmol/L. Except IL-12, the expression of other 5 cytokines (IL-6, TNF-α, IL-1β, IL-2, IL-8) was significantly inhibited by 1.6 μmol/L SHR0302 in SET2 cells at 24 h (P<0.01) , while Ruxolitinib 1.0 μmol/L had the same effect. Several phosphorylated molecules of Jak-Stat signaling pathway were significantly inhibited by SHR0302 in SET2 cells only for 3 h. P-stat1 (Tyr701) , p-stat3 (Tyr705) were down-regulated when treated with SHR0302 1.0 μmol/L (P<0.05) , p-jak1 (tyr1022/1023) and p-stat5 (Tyr694) were inhibited at 5.0 μmol/L (P<0.05) . Ruxolitinib significantly inhibited the downstream STAT protein at 0.1 μmol/L. Again, the inhibitory effect of SHR0302 on protein expression was weaker than that of Ruxolitinib.@*Conclusion@#SHR0302 can effectively inhibit the proliferation of MPN cell line and patients' primary cells, as well as the expression of inflammatory factors. The molecular mechanism is possibly related to the down-regulation of phosphorylated proteins of Jak-Stat signaling pathway. Overall, the anti-proliferative and anti-inflammatory effects of SHR0302 are weaker than those of Ruxolitinib.
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Acute respiratory distress syndrome (ARDS) is an acute pulmonary edema induced by non-cardiac factors and a clinical syndrome characterized by respiratory distress and refractory hypoxemia. The pathogenesis of ARDS is complex. Systemic or local damaging factors can aggravate the inflammatory injury of lung tissue dependent on the activation of endoplasmic reticulum stress (ERs) and unfolded protein responses (UPR) and correlation with various damaging mechanisms such as inflammatory response, oxidative stress, apoptosis, autophagy, and calcium homeostasis. This article reviews the progress of ERs associated with ARDS to help us understand the pathogenesis of ARDS.
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Acute respiratory distress syndrome (ARDS) is an acute pulmonary edema induced by non-cardiac factors and a clinical syndrome characterized by respiratory distress and refractory hypoxemia. The pathogenesis of ARDS is complex. Systemic or local damaging factors can aggravate the inflammatory injury of lung tissue dependent on the activation of endoplasmic reticulum stress (ERs) and unfolded protein responses (UPR) and correlation with various damaging mechanisms such as inflammatory response, oxidative stress, apoptosis, autophagy, and calcium homeostasis. This article reviews the progress of ERs associated with ARDS to help us understand the pathogenesis of ARDS.
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Objective@#To evaluate clinical characteristics and prognosis of primary myelofibrosis (PMF) patients with thrombocytopenia in varied degrees.@*Methods@#Clinical features and survival data of 1 305 Chinese patients with PMF were retrospectively analyzed. The prognostic value of thrombocytopenia in patients with PMF was evaluated.@*Results@#320 subjects (47%) presented severe thrombocytopenia (PLT<50×109/L), 198 ones (15.2%) mild thrombocytopenia [PLT (50-99)×109/L] and 787 ones (60.3%) without thrombocytopenia (PLT ≥ 100×109/L). The more severe the thrombocytopenia, the higher the proportions of HGB<100 g/L, WBC<4×109/L, circulating blasts ≥ 3%, abnormal karyotype and unfavourable cytogenetics (P<0.001, P<0.001, P=0.004, P<0.001 and P<0.001, respectively) were observed in this cohort of patients. The more severe the thrombocytopenia, the lower the proportion of JAK2V617F positive (P<0.001) was also noticed. Platelet count was positively correlated with splenomegaly, HGB and WBC (P<0.001, correlation coefficients were 0.131, 0.445 and 0.156, respectively). Platelet count was negative correlated with constitutional symptoms and circulating blasts (P=0.009, P=0.045, respectively; correlation coefficients were -0.096 and -0.056, respectively). The median survival of patients with severe thrombocytopenia, mild thrombocytopenia and without thrombocytopenia were 32, 67 and 89 months, respectively (P<0.001). Multivariate analysis identified thrombocytopenia in varied degrees (HR=1.693, 95%CI 1.320-2.173, P<0.001) and Dynamic Internation Prognostic Scoring System(DIPSS) prognostic model (HR=2.051, 95%CI 1.511-2.784, P<0.001) as independent risk factors for survival.@*Conclusion@#PMF patients with severe thrombocytopenia frequently displayed anemia, leucopenia, circulating blasts and short survival, so active treatment measures should be taken especially in these patients.
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Objective BRCA1 is one of the most important susceptibility genes of breast cancer. The article aimed to investigate the expression of BRCA1 and its correlation in sporadic invasive breast cancer. Methods The expressions of BRCA1, ER, PR, HER2 and Ki67 in 618 cases of sporadic invasive breast cancer in our hospital from January 2015 to December 2017 were detected with immunohistochemistry in order to investigate and analyze the expression of BRCA1 and its correlation with molecular classification, histological type and other related molecular markers in sporadic invasive breast cancer. Results The positive rate of BRCA1 was 44.2% consisting of 30.3% weak positive(+) and 13.9% strong positive(++). The positive rates of ER, PR, and HER2 are 60.8%, 54.7%,and 24.9%. The proliferation index ≤30% of Ki67 was 70.7%, >30% was 29.3%. The expression of BRCA1 in luminal type A was significantly lower than the other four types of sporadic invasive breast cancer(P<0.05). The expression of BRCA1 in breast cancer with medullary histological features was significantly lower than those of the other types of breast cancer(P<0.05). There was significant difference between the expression of BRCA1 and the expressions of ER, HER2 and Ki67 (P<0.01). The expression of BRCA1 had positive correlation with expression of HER2 in sporadic invasive breast cancer (r=0.117,P<0.01). Conclusion The expression of BRCA1 in sporadic invasive breast cancer with triple negative subtype and medullary histological features is down-regulated and BRCA1 may affect the development and progression of sporadic invasive breast cancer.
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Objective@#To investigate the predictive value of neutrophil-to-lymphocyte Ratio (NLR) in 30-day mortality of patients with acute paraquat poisoning.@*Methods@#We respectively reviewed the clinical parameters of 115 patients with acute paraquat poisoning. They were divided into survival (n=64) and non-survival (n=51) groups based on their 30-day outcome. Multivariate logistic regression was performed to identify risk factors of 30-day mortality. Receiver operating curve (ROC) test was applied to analysis to the predictive value of NLR in 30-day mortality ofacute paraquat poisoning patients. The correlations between NLR and severity index of paraquat poisoning (SIPP) were analyzed using Spearman’s rank correlation coefficient.@*Results@#Of the 115patients included in the study, 54 (46.96%) patients were males and 61 (53.04%) were females with a mean age of 38.96±13.58 years. The total mortality in 30-day was 44.35% (51/115) . The NLR at admission was an independently risk factor of 30-day mortality of patients with acute paraquat poisoning (OR 1.477, 95%CI 1.035-2.107, P<0.05) . The NLR to predict the death of the area under the ROC curve was 0.894 (95%CI: 0.8212-0.9663, P<0.01) ; the optimal cutoff threshold was 11.71; the sensitivity was 71.79% and the specificity was 94.29%; the positive predictive value was 93.33%and negative predictive value of 75.00%. Meanwhile, the positive likelihood ratio was 12.57 and the negative likelihood ratio was 0.30. The NLR was significantly associated with SIPP (Spearman rho 0.525; P<0.01) and it was significantly higher in patients with SIPP of ten or higher than in those with an SIPP less than 10 (15.02±12.40 vs. 6.19±2.54, P<0.05) .@*Conclusion@#The increased NLR at admission was an independently risk factor of 30-day mortality of patients with acute paraquat poisoning and it was significantly correlated with SIPP score. Therefore, NLR was useful for predicting prognosis of patients with acute paraquat poisoning.
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Objective To investigate the clinical features and prognosis risk factors of the elderly and youth patients with acute severe poisoning.Methods Adult patients with acute severe poisoning in the emergency intensive care unit (EICU) of Nanjing Drum Tower Hospital from January 2008 to December 2017 were enrolled. The patients were divided into the elderly group (age ≥ 60 years) and the youth group (16 years≤age < 60 years), the clinical data of the two groups were analyzed. The patients were divided into survival group and death group according to the prognosis of 28-day, binary multivariate Logistic regression was used to analyze the risk factors of mortality of the elderly and youth patients; receiver operating characteristic curve (ROC) was used to assess the predictive value of acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) in mortality of youth patients.Results A total of 343 patients with acute severe poisoning were included, 89 in the elderly group and 254 in the youth group. ① Clinical features: compared with the youth group, the elderly group had higher proportion of basic diseases included hypertension, type 2 diabetes and coronary heart disease, higher the initial APACHEⅡ scores at admission, higher the proportion of invasive mechanical ventilation and respiratory failure, and longer the length of EICU stay and the length of hospital stay. The main poisoning causes of elderly and youth patients were suicide (58.43％, 83.86％) and accidents (38.20％, 13.39％). The most common poisoning types of elderly patients were sedative hypnotics (23.60％) and organophosphorus pesticides (22.47％); the youth patients were mainly paraquat (42.52％) and organophosphorus pesticide (17.32％). There were 28 patients died (31.46％) in the elderly group and the cause of death were respiratory failure (53.57％), circulatory failure (32.14％) and multiple organ dysfunction syndrome (MODS, 14.29％). There were 67 patients died (26.38％) in the youth group and the cause of death were respiratory failure (59.70％), MODS (20.90％) and circulatory failure (19.40％). ② Risk factors of deaths: the APACHEⅡ score, incidence of acute kidney injury (AKI) and MODS in the elderly death group were significantly higher than those in the elderly survival group. Logistic regression analysis showed that AKI was the independent risk factor for death in elderly patients [odds ratio (OR) = 8.449, 95％ confidence interval (95％CI) =2.347-30.410,P = 0.001]. The proportion of female, APACHE Ⅱ score, and the incidence of AKI, respiratory failure and MODS in the youth death group were significantly higher than those in the youth survival group. Logistic regression analysis showed that APACHE Ⅱ score (OR = 1.175, 95％CI = 1.081-1.277,P = 0.001), AKI (OR = 34.470, 95％CI =11.681-101.722,P = 0.001) and MODS (OR = 3.834, 95％CI = 1.264-11.636,P = 0.018) were the independent factors for death in the youth patients. ③ Predictive value: the initial APACHEⅡscore was useful for predicting prognosis of youth patients with acute severe poisoning. The APACHE Ⅱ score to predict the death of the area under the ROC curve (AUC) was 0.744 (95％CI = 0.681-0.806,P = 0.001); the cut-off was 5, the sensitivity was 92.54％, the specificity was 51.34％, the positive predictive value was 65.53％, the negative predictive value was 87.31％, the positive likelihood ratio was 1.902, and the negative likelihood ratio was 0.145.Conclusions Patients with acute severe poisoning have their own clinical characteristics. To reduce the morbidity and improve the prognosis, we should strengthen the pre-hospital management and optimize the clinical treatment process.
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Objective To observe the analgesic effect of compound coeruleum methylene blue preparation after thoracoscopic surgery,and to provide a new idea for the choice of analgesic drugs for thoracoscopic patients.Methods The clinical data of 116 patients who underwent selective thoracoscopic surgery in the Department of Cardiothoracic Surgery of Jieyang People's Hospital from January 2016 to May 2018 were analyzed.This study was a prospective study.Research subjects were divided into control group (n =57) and observation group (n =59) by random digital table.In the two groups,no analgesic measures were used in advance,of which the patients in the control group were given the routine injection of piperidine hydrochloride by intramuscular injection after operation.While patients in the observation group were given intercostal nerve block across the upper and lower fibs after operation,and the dosage of compound methylene blue was 1 ml in each rib.on the basis with pethidine hydrochloride.The visual analog score and Ramsay sedation score were compared between the two groups of patients at 4 h,12 h 24 h and 48 h after operation and compared the use of pethidine hydrochloride,adverse drug reactions,ambulation time and hospitalization time.The counting data was expressed in terms of frequency and percentage (％).The measurement data were represented by ((x) ± s).The comparison of different time points within the group was performed by single-factor analysis of variance,and the comparison between the two groups was performed by independent sample t test.Results Compared with the control group's patients [in resting state,12 h:(3.85 ± 1.97) points;24 h:(2.74 ± 1.91) points;48 h:(2.11 ± 1.70) points;in activity state,12 h:(5.02 ± 1.64) points;24 h:(4.89 ± 1.36) points;48 h:(3.83 ± 1.51) points],the VAS scores of the patients in the observation group in resting state [12 h:(2.68 ± 1.24) points;24 h:(1.35 ± 1.16) points;48 h:(0.74 ± 0.63) points] and activity state[12 h:(3.81 ±1.53) points;24 h:(3.25±1.71) points;48 h:(2.42± 1.33)points] were significantly lower at 12 h,24 h and 48h after operation.Compared with the control group's patients 12 h [(2.58 ± 0.41) points] and 24 h [(2.29 ± 0.34) points],the Ramsay scores were significantly higher at 12 h[(2.93 ±0.35)points] and 24 h [(2.79 ±0.30)points] after operation (P＜0.05).The observation group's amount of piperidine hydrochloride was less,and the ambulation and hospitalization time were shorter than the control group's (P ＜ 0.05).The incidence of analgesia-related adverse reactions in the control group and the observation group were 35.09％ and 27.12％,respectively,with no significant difference (P ＞ 0.05).Conclusions Compound coeruleum methylenum's analgesic effect after thoracoscopic surgery is accurate and maintains long time,and the side effects are less and the operation is convenient.It is worthy of clinical application.
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Acute ischemic stroke (AIS), as the third leading cause of death worldwide, is characterized by its high incidence, mortality rate, high incurred disability rate, and frequent reoccurrence. The neuroprotective effects of Ginkgo biloba extract (GBE) against several cerebral diseases have been reported in previous studies, but the underlying mechanisms of action are still unclear. Using a novel in vitro rat cortical capillary endothelial cell-astrocyte-neuron network model, we investigated the neuroprotective effects of GBE and one of its important constituents, Ginkgolide B (GB), against oxygen-glucose deprivation/reoxygenation and glucose (OGD/R) injury. In this model, rat cortical capillary endothelial cells, astrocytes, and neurons were cocultured so that they could be synchronously observed in the same system. Pretreatment with GBE or GB increased the neuron cell viability, ameliorated cell injury, and inhibited the cell apoptotic rate through Bax and Bcl-2 expression regulation after OGD/R injury. Furthermore, GBE or GB pretreatment enhanced the transendothelial electrical resistance of capillary endothelial monolayers, reduced the endothelial permeability coefficients for sodium fluorescein (Na-F), and increased the expression levels of tight junction proteins, namely, ZO-1 and occludin, in endothelial cells. Results demonstrated the preventive effects of GBE on neuronal cell death and enhancement of the function of brain capillary endothelial monolayers after OGD/R injury in vitro; thus, GBE could be used as an effective neuroprotective agent for AIS/reperfusion, with GB as one of its significant constituents.
Subject(s)
Animals , Rats , Apoptosis , Brain Ischemia , Drug Therapy , Cell Survival , Cells, Cultured , Disease Models, Animal , Endothelial Cells , Ginkgolides , Pharmacology , Glucose , Lactones , Pharmacology , Neurons , Neuroprotective Agents , Pharmacology , Oxygen , Plant Extracts , Pharmacology , Stroke , Drug TherapyABSTRACT
A study of 426 rabbits from 3 cities in Jilin province (Changchun City and Jilin City) and Liaoning province (Shenyang City) was conducted between May and June 2015. The overall prevalence of E. bieneusi in rabbits was 0.94% (4/426), with 0% (0/116), 1.72% (3/174), and 0.74% (1/136) in Jilin, Changchun, and Shenyang City, respectively. Only 3 farms (farm 1 and farm 3 in Changchun City, farm 8 in Shenyang City) were PCR-positive for E. bieneusi. Moreover, rabbits of more than 6 months (1.72%) had the highest E. bieneusi prevalence, followed by rabbits of 4-6 months (1.26%), 2-3 months (0.58%), and less than 1 month (0%). Analysis of ITS gene of E. bieneusi suggested that all 4 E. bieneusi isolates were genotype D, and were classified as group 1a. The present results first demonstrated the existence of zoonotic E. bieneusi in domestic rabbits in China. Effective control measures should be implemented to prevent E. bieneusi infection in domestic rabbits, other animals, and humans.
Subject(s)
Animals , China/epidemiology , DNA, Ribosomal Spacer/genetics , Enterocytozoon/genetics , Genotype , Microsporidiosis/epidemiology , Rabbits/microbiology , Zoonoses/microbiologyABSTRACT
Serine proteases form one of the most important families of enzymes and perform significant functions in a broad range of biological processes, such as intra- and extracellular protein metabolism, digestion, blood coagulation, regulation of development, and fertilization. A number of serine proteases have been identified in parasitic helminths that have putative roles in parasite development and nutrition, host tissues and cell invasion, anticoagulation, and immune evasion. In this review, we described the serine proteases that have been identified in parasitic helminths, including nematodes (Trichinella spiralis, T. pseudospiralis, Trichuris muris, Anisakis simplex, Ascaris suum, Onchocerca volvulus, O. lienalis, Brugia malayi, Ancylostoma caninum, and Steinernema carpocapsae), cestodes (Spirometra mansoni, Echinococcus granulosus, and Schistocephalus solidus), and trematodes (Fasciola hepatica, F. gigantica, and Schistosoma mansoni). Moreover, the possible biological functions of these serine proteases in the endogenous biological phenomena of these parasites and in the host-parasite interaction were also discussed.
Subject(s)
Animals , Cestoda/classification , Host-Parasite Interactions , Life Cycle Stages , Nematoda/classification , Serine Proteases/genetics , Trematoda/classificationABSTRACT
<p><b>OBJECTIVE</b>To analyze ginsenosides composition in wild ginseng leaves with different growth years.</p><p><b>METHOD</b>The analysis was performed on Acquity UPLC BEH Shield RP18 (2.1 mm x 100 mm, 1.7 microm) column, the mobile phase was acetonitrile-0.05% formic acid solution in gradient elution mode. The detection wavelength was at 203 nm. The flow rate was 0.4 mL x min(-1) and column temperature was set at 30 degrees C.</p><p><b>RESULT</b>Thirteen ginsenosides were determined by the established UPLC method. In 5-17th growth year ginseng leaf samples cultivated simulating wild conditions, the contents of ginsenosides in the 14th year have the highest content.</p><p><b>CONCLUSION</b>The established method is simple, accurate and reliable, can be used in ginsenosides determination and fingerprint research of Panax crude drug. The result provides reliable data for the accumulation of ginsenosides and sustainable utilization of ginseng resources.</p>
Subject(s)
Drugs, Chinese Herbal , Ginsenosides , Panax , Chemistry , Plant Leaves , Chemistry , Time FactorsABSTRACT
Acetylcholinesterase (AChE) inhibitors are mainly used in the treatment of Alzheimer's disease (AD). The inhibitory effect of icariin on the activity of AChE was investigated by inhibition kinetics. The binding interaction and binding sites between icariin and AChE were also studied by using fluorimetry and molecular docking, respectively. The results showed that icariin could potently inhibit the activity of AChE, the IC50 value was determined to be 3.50 x 10(-8) mol x L(-1), and the determined IC50 value to tacrine was 0.75 x 10(-8) mol x L(-1). Kinetic analyses showed that icariin is a reversible and mixed type AChE inhibitor. The inhibition constants K1 and K(IS) were determined to be 2.67 x 10(-8) and 4.43 x 10(-8) mol x L(-1), respectively. Icariin binds selectively to the AChE peripheral anionic site via hydrogen bonds and Van der Waals forces.
Subject(s)
Acetylcholinesterase , Metabolism , Binding Sites , Cholinesterase Inhibitors , Pharmacology , Drugs, Chinese Herbal , Pharmacology , Epimedium , Chemistry , Flavonoids , Pharmacology , Hydrogen Bonding , Inhibitory Concentration 50 , Kinetics , Molecular Docking Simulation , Plants, Medicinal , ChemistryABSTRACT
<p><b>BACKGROUND</b>Adipose-derived stromal cell (ADSC) differentiation into neural cells in vitro is becoming widely studied. However, there are few reports on astrocytes following differentiation, and particularly on maturation and electrophysiology. In this study, we used various methods to determine ADSC-derived astrocyte maturity.</p><p><b>METHODS</b>Chemical induction with isobutylmethylxanthine (IBMX) was used to differentiate adult ADSCs into astrocytes followed by hematoxylin-eosin (HE) staining to observe morphology and transmission electron microscopy for cellular ultrastructure assessment. Immunofluorescence was used to detect expression of neural stem cell marker nestin as well as glial markers glial fibrillary acidic protein (GFAP) and S-100. In addition, we measured membrane potentials in bis-(1,3-dibarbituric acid) trimethine oxanol-labeled ADSCs and astrocytes by stimulation with a high potassium solution under an inverted fluorescence microscope. Finally, cell cycle distribution was detected by flow cytometry.</p><p><b>RESULTS</b>Typical astrocyte morphology was shown by HE staining after 48-hour differentiation. Glial fibril was observed with transmission electron microscopy. GFAP and S-100 were not expressed in the control group, but were expressed within 24-hour differentiation and reached a maximum at day 14 with no change up to day 28. Nestin was weakly expressed in control cells and also reached a maximum at day 14 with the percentage of positive cells constant until day 21 followed by a decrease. Differentiated cell membrane potentials after stimulation with potassium were slightly increased, and then gradually declined over time. There was no significant membrane potential change in the control group. Flow cytometry showed that the percentage of cells in G0/G1 phase was 93% and only 5% in S phase.</p><p><b>CONCLUSION</b>ADSCs were differentiated into mature astrocytes with typical characteristics including morphology, ultrastructure, marker protein expression, mature potassium channels and mitotic capacity.</p>
Subject(s)
Adult , Female , Humans , Male , Young Adult , 1-Methyl-3-isobutylxanthine , Pharmacology , Adipose Tissue , Cell Biology , Astrocytes , Cell Biology , Barbiturates , Pharmacology , Cell Differentiation , Cells, Cultured , Electrophysiology , Methods , Flow Cytometry , Glial Fibrillary Acidic Protein , Metabolism , Membrane Potentials , Microscopy, Fluorescence , S100 Proteins , Metabolism , Stromal Cells , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate whether cyclooxygenase-2 (COX-2) and heme oxygenase-1 (HO-1) are involved in the bradykinin-induced delayed protection.</p><p><b>METHODS</b>Cardiac contractility, lactate dehydrogenase (LDH) and infarct area were analyzed in isolated rat hearts undergoing ischemia-reperfusion injury induced by Langendorff method.</p><p><b>RESULT</b>Conscious rats received bradykinin (40 microg/kg), and the isolated hearts were subjected to 30 min of regional ischemia and 120 min of reperfusion 24 h later. Bradykinin pretreatment would improve post-ischemic performance, and reduced the release of LDH and infarct size. COX-2 inhibitor celecoxib (3 mg/kg) abolished bradykinin-induced protection, leading to poorer myocardial performance, release of more LDH and larger infarct sizes. Administration of HO-1 inhibitor ZnPP IX(20 microg/kg) before bradykinin partially abrogated the delayed protection. Pretreatment with the mitochondrial ATP sensitive potassium channel(mitoK(ATP) antagonist 5-HD before or 24 h after bradykinin administration also abolished the effect of protection.</p><p><b>CONCLUSION</b>The results indicate that activation of HO-1 and COX-2 might be involved in the delayed cardioprotection evoked by bradykinin, and mitoK(ATP) channel may serve as both a trigger and a mediator in the cardioprotection.</p>