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Objective:To explore the clinical characteristics of Mycoplasma pneumoniae (MP) bronchiolitis to provide references for clinical prevention and treatment.Methods:From January 2019 to January 2020, the clinical data of children diagnosed with MP bronchiolitis in the Department of Respiratory 2 of Hebei Children′s Hospital were retrospectively collected and analyzed, and the patients were followed up to observe the prognosis.Results:Among the 67 children, there were 39 boys and 28 girls, with age of 5 (1-14) years.All the children had cough, 63 cases (94.0%) had fever, 49 cases (77.8%) had high fever (≥39.0 ℃), and the median total fever course was 8(1-27) d; 10 cases (14.9%) had wheezing; 5 cases (7.5%) had dyspnea; 3 cases (4.5%) had hypoxemia; wet rales were heard in 58 cases (86.6%) and wheezing in 10 cases (14.9%). Forty-eight patients (71.6%) had family allergy history or personal allergy history.The average value of peripheral blood white blood cells was (8.6±2.6)×10 9/L; the me-dian of C reactive protein was 9.1(0.5-55.6) mg/L, of which 7 cases (10.4%) were ≥30 mg/L; the median of lactate dehydrogenase was 278(181-590) U/L.Tree bud sign and central lobular nodules were the main findings of high-resolution CT of the chest.Among them, 45 cases (67.2%) had bilateral lesions, 38 cases (56.7%) involved ≥3 lung lobes, and 22 cases (32.8%) had a little lung consolidation, 19 cases (28.4%) had a small amount of pleural effusion.The bronchoscopic features of 35 children (52.2%): 71.4%(25/35 cases) showed white flocculent and cord-like secretions, 17.1%(6/35 cases) displayed mucus plugs blocking the lumen, and 10.9%(4/35 cases) showed no-dular protrusions and granulation tissue hyperplasia.All children were treated with Azithromycin sequentially; 42 children (62.7%) were treated with low-dose hormones, and the average initiation time was on day (10.62±2.71) and the median course was 14(2-42) d. Sixty-seven cases were followed up for half a year, of which 61 cases (91.0%) had almost complete lung recovery, and 6 cases (9.0%) developed bronchiolitis obliterans (BO). Conclusions:MP bronchiolitis mostly occurs in infants and preschool children with a family history of allergies or personal allergies.Fever and cough are the main clinical manifestations.In severe cases, dyspnea and hypoxemia may occur.The physical signs of the lungs are mainly include wet rales, some of which are wheezing.White flocculent and cord-like secretions are the main manifestations of bronchoscopy.After active treatment, most children have a good prognosis, and some children may develop BO.
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Objective To investigate the effects of ephedrine on the expression levels of brain-derived neurotropic factor (BDNF) and postsynaptic density protein 95 (PSD95) and synapsin1 in PC12 cells, and to explore the mechanism of ephedrine cytotoxicity on PC12. Methods After PC12 cells were treated with different concentration of ephedrine, the cell survival rate was measured by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The morphology changes of PC12 cells were observed by an inverted microscope. Western blot was used to detect the protein expression levels of BDNF, PSD95 and synapsin1 in PC12 cells. Results Ephedrine decreased the viability of PC12 cell in a concentration-dependent manner,with an IC25 and IC50 of 0.536 mmol and 2.8 mmol, respectively, for PC12 cell death. As ephedrine concentration increased, PC12 cells became smaller in size, with blurred boundary blurred, reduced synapses and shorter axon lengths. The expression levels of BDNF and PSD95 increased significantly. Meanwhile the expression level of synapsin1 decreased. Conclusion The mechanism of ephedrine cytotoxicity on PC12 may be related to the expression levels of BDNF, PSD95 and synapsin1.
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OBJECTIVE@#To study the epidemiological features of children with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Shijiazhuang, China.@*METHODS@#Based on the information officially announced on the official website of the Health Commission of Hebei Province, epidemiological data were collected from 133 children, aged 0-18 years, who were diagnosed with SARS-CoV-2 infection in Shijiazhuang from January 2 to January 30, 2021. A statistical analysis was performed for general status, regional distribution, presence or absence of clusters, and results of SARS-CoV-2 nucleic acid tests.@*RESULTS@#Among the 133 children with SARS-CoV-2 infection, there were 65 boys and 68 girls, with a male/female ratio of 0.96:1. The youngest age of onset was 3 months and 7 days, and the mean age of onset was (9±5) years. Of all the 133 children, 90(67.7%) were the first confirmed case of SARS-CoV-2 infection among their family members. Of all the children, 108(81.2%) came from the Gaocheng District in Shijiazhuang, among whom 38(28.6%) were from Xiaoguozhuang Village where the first patient with a confirmed diagnosis lived. SARS-CoV-2 nucleic acid test at week 2 after the outbreak showed positive results in 88 children (66.2%), and only 5 children had clinical symptoms before positive SARS-CoV-2 results were obtained. Of all the 133 children, 19(14.3%) were found positive in the first SARS-CoV-2 nucleic acid test after the outbreak, and 70(52.6%) had positive results for ≥4 times. There were 98 school students with infection, among whom 74(75.5%) were the first confirmed case in their family, and among 35 non-school students, 16(45.7%) were the first confirmed case in their family (@*CONCLUSIONS@#Among the children confirmed with SARS-CoV-2 infection in Shijiazhuang, there is a high proportion of children who are the first confirmed case in their family, and the children are mainly distributed in the rural areas of Gaocheng. Most of these children are students, so the prevention and control of cluster infection in schools should be taken seriously. There are often no symptoms before SARS-CoV-2 nucleic acid test, with a low positive rate of the first nucleic acid test, which increases the difficulty of early discovery of the epidemic.
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Adolescent , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , COVID-19 , China/epidemiology , Disease Outbreaks , SARS-CoV-2 , SchoolsABSTRACT
The concept of total mesorectal resection provides a quality control standard that can be followed for radical resection of rectal cancer, but some anatomical problems are still controversial. Compared with traditional open surgery, laparoscopic radical rectal surgery has better surgical vision, better neurological protection, better operating space. However, if the surgeon has insufficient understanding of the anatomy, collateral damage may occur, such as uncontrollable bleeding during the operation, postoperative urination and defecation dysfunction and so on. Based on the interpretation of the researches at home and abroad, combined with the clinical experience, we elucidate some associated issues, including anatomic variation of inferior mesenteric vessels, the controversy of inferior mesenteric artery ligation plane, the controversy of lymph node dissection in No. 253, the anatomical variation of middle rectal artery, and the anatomical controversy of lateral lymph node dissection in rectal cancer, in order to provide better cognitive process for the clinical front-line surgeons.
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Humans , Laparoscopy , Lymph Node Excision , Lymph Nodes , Mesenteric Artery, Inferior , Rectal Neoplasms/surgery , RectumABSTRACT
In December 2019, a new outbreak of coronavirus pneumonia began to occur. Its pathogen is 2019-nCoV, which has the characteristics of strong infectivity and general susceptibility. The current situation of prevention and control of new coronavirus pneumonia is severe. In this context, as front-line medical workers bearing important responsibilities and pressure, while through strict management strategy, we can minimize the risk of infection exposure. By summarizing the research progress and guidelines in recent years in the fields of colorectal cancer disease screening, treatment strategies(including early colorectal cancer, locally advanced colorectal cancer, obstructive colorectal cancer, metastatic colorectal cancer and the treatment of patients after neoadjuvant therapy), the choice of medication and time limit for adjuvant therapy, the protective measures for patients undergoing emergency surgery, the re-examination of postoperative patients and the protection of medical staff, etc., authors improve treatment strategies in order to provide more choices for patients to obtain the best treatment under the severe epidemic situation of new coronavirus pneumonia. Meanwhile we hope that it can also provide more timely treatment modeling schemes for colleagues.
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Objectives To analyze the clinical characteristics, treatment and prognosis of necrotizing pneumonia caused by Mycoplasma pneumoniae (MP) infection in children. Method The clinical data of children with necrotizing pneumonia cause by MP infection from October 2016 to October 2017 were retrospectively analyzed. Results A total of 26 children (10 males and 16 females) with an average age of (5.76±2.60) years, were enrolled in the study. All children were characterized by fever and cough. High fever ( ≥ 39.0 ℃) was seen in 23 cases (88.5％) and the total duration of fever was (16.88±7.42) days. Pulmonary auscultation showed a reduction in respiratory sounds in all children. The range of peripheral blood leukocytes were (9.0～36.8) ×109/L, mean peak neutrophil ratio was (69.2±13.2) ％, and the range of C-reactive protein (CRP) was (1～202.5) mg/L. The mean value of lactic dehydrogenase (LDH) was (448±247) U/L. At the beginning of the disease, the chest images showed homogeneous solid high-density images over the whole lung lobe and 20 cases (76.9％) were complicated with pleural effusion. At the later stage, lung CT showed thin-walled cavities or multiple air-containing cysts on the basis of lung consolidation. Fiberoptic bronchoscopy showed lumen obstruction caused by mucus plugs in 23 cases (88.5％) . All the children were treated with methylprednisolone. The dose of 2 mg/ (kg·d) was effective in 21 cases and the fever was relieved in 5 cases after the dose was adjusted to 4 mg/ (kg·d) , and the average hormone application time was (13.08 ± 8.38) d. The median length of hospital stay was [16.5 (7～32) ] d. Two cases were lost to follow-up and 24 cases finished 6-month follow-up. Lung CT showed almost complete recovery of the lungs in 16 cases, residual pleural hypertrophy in 5 cases, and bronchiectasis in 1 case and bronchiolitis obliterans in 2 cases. Conclusion Necrotic pneumonia in children caused by MP infection is characterized by persistent high fever, decreased respiratory sounds, lung consolidation and mucus plugs induced lumen obstruction. The prognosis is relatively good after active anti-infection and hormone therapy.
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Objective@#To investigate the epidemiologic features of respiratory viral etiology in hospitalized children with acute respiratory tract infection (ARTI) in Shijiazhuang.@*Methods@#A total of 28 512 cases of hospitalized children with clinical diagnosis of ARTI in Children′ s Hospital of Hebei Province from 2014 to 2017 were recruited into this study. One nasopharyngeal swab was collected from each patient. Immunofluorescence assay was used to detect seven kinds of respiratory viruses, including respiratory syncytial virus (RSV), parainfluenza virus (PIV) type 1-3, influenza virus type A, B (FluA, FluB) and adenovirus (ADV).@*Results@#At least one viral pathogen was identified in each of 9 263 out of 28 512 patients and the overall positive rate was 32.5%. Of 9 263 virus-positive patients, 9 070 (97.9%) had mono-infection. The most frequently detected virus was RSV, followed by PIV-3 and FluA. The positive rates of RSV and PIV-1 showed annually decreasing tendency, meanwhile the positive rate of FluA increased in the nearly two years. The detection rate of ADV and PIV-1 increased every other year. There was a significant difference in the positive rate among different years (P<0.05). The overall positive rate decreased along with the age increased (linear by linear association χ2=1191.289, P<0.05). The detection rates of RSV and PIV-3 were the highest in groups of <1 year old and 1-3 years old and decreased along with the age increased. The preschool children were more susceptible to developing FluA, FluB and ADV related diseases. There was a significant difference in the positive rate among different age groups (P<0.05). The viral distribution was uneven in different seasons, and the infection peaked in winter, the difference was statistically significant (P<0.05). The epidemic seasons of RSV and FluA were winter, and FluB infection was epidemic in winter and spring. The positive rates of PIV-1 and PIV-2 were most common in summer and autumn. PIV-3 was usually prevalent in spring and summer and ADV was prevalent sporadically.@*Conclusions@#RSV is the most common pathogen in hospitalized children with clinical diagnosis of ARTI during 2014-2017 and the positive rate of which showed an annually decreasing tendency. The positive rate of FluA increased in the nearly two years. Children in infancy are susceptible to the seven common respiratory viruses and winter is the epidemic season for these viruses.
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<p><b>OBJECTIVE</b>To study the efficacy of early treatment via fiber bronchoscope in children with Mycoplasma pneumoniae pneumonia (MPP) complicated by airway mucus obstruction.</p><p><b>METHODS</b>According to the time from admission to the treatment via fiber bronchoscope, the children with MPP who were found to have airway mucus obstruction under a fiber bronchoscope were randomly divided into early intervention group (≤3 days; n=40) and late intervention group (>3 days; n=56). The two groups were compared in terms of clinical data and imaging recovery.The children were followed for 1-3 months.</p><p><b>RESULTS</b>Of the 96 children, 38 were found to have the formation of plastic bronchial tree, among whom 10 were in the early intervention group and 28 were in the late intervention group (P=0.01). Compared with the late intervention group, the early intervention group had a shorter duration of fever, length of hospital stay, and time to the recovery of white blood cell count and C-reactive protein (P<0.05), as well as a higher atelectasis resolution rate (P<0.05). Compared with the late intervention group, the early intervention group had a higher percentage of children with a ≥ 60% absorbed area of pulmonary consolidation at discharge. After 3 months of follow-up, the early intervention group had a higher percentage of children with a ≥ 90% absorbed area of pulmonary consolidation than the late intervention group (80% vs 55%; P=0.01), and the early intervention group had a lower incidence rate of atelectasis than the late intervention group (P<0.05).</p><p><b>CONCLUSIONS</b>Early treatment via fiber bronchoscope can shorten the course of the disease and reduce complications and sequelae in MPP children with airway mucus obstruction.</p>
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Child , Child, Preschool , Female , Humans , Male , Airway Obstruction , Therapeutics , Bronchoscopes , Fiber Optic Technology , Mucus , Pneumonia, MycoplasmaABSTRACT
BACKGROUND AND PURPOSE: Vasa vasorum (VV) have been believed to be rare or non-existent in small-caliber intracranial arteries. In a series of human cerebral artery specimens, we identified and examined the distribution of VV in association with co-existing intracranial atherosclerosis. METHODS: We obtained cerebral artery specimens from 32 consecutive autopsies of subjects aged 45 years or above. We scrutinized middle cerebral artery (MCA), vertebral artery (VA), and basilar artery (BA) for the presence of adventitial VV. We described the distribution of VV, and the characteristics of co-existing atherosclerotic lesions. RESULTS: Among 157 intracranial arteries, adventitial VV were present in 74 of the 157 specimens (47%), involving MCA (n=13, 18%), BA (n=14, 19%), and VA (n=47, 64%). Although qualitatively these 74 adventitial VV distributed similarly in arteries with or without atherosclerotic lesions (disease-free arteries n=4/8; arteries of pre-atherosclerosis n=17/42; and arteries of progressive atherosclerosis n=53/107), the presence of adventitial VV in intracranial VA was associated with a heavier plaque load (1.72±1.66 mm2 vs. 0.40±0.32 mm2, P < 0.001), severer luminal stenosis (25%±21% vs. 12%±9%, P=0.002), higher rate of concentric lesions (79% vs. 36%, P=0.002), and denser intraplaque calcification (44% vs. 0%, P=0.003). Histologically, intracranial VA with VV had a larger diameter (3.40±0.79 mm vs. 2.34±0.58 mm, P < 0.001), thicker arterial wall (0.31±0.13 mm vs. 0.23±0.06 mm, P=0.002), and a larger intima-media (0.19±0.09 mm vs. 0.13± 0.04 mm, P=0.003) than VA without VV. CONCLUSIONS: Our study demonstrated the distribution of adventitial VV within brain vasculature and association between vertebral VV and progressive atherosclerotic lesions with a heavier plaque load and denser intraplaque calcification.
Subject(s)
Humans , Arteries , Atherosclerosis , Autopsy , Basilar Artery , Brain , Cerebral Arteries , Constriction, Pathologic , Intracranial Arteriosclerosis , Middle Cerebral Artery , Phenobarbital , Vasa Vasorum , Vertebral ArteryABSTRACT
BACKGROUND: Cell-free stem cell therapy has been an issue of concern, but there is no conclusion on how to extract high-quality exosomes. OBJECTIVE: To extract exosomes from human umbilical cord mesenchymal stem cells by using three different methods, and then to screen the optimal method. METHODS: Exosomes were extracted from human umbilical cord mesenchymal stem cells by using the Total Exosome Isolation test kit, Exo Quick test kit and differential ultracentrifugation method, respectively. Then, transmission electron microscopy was used for morphological observations, BCA was utilized to quantify the protein, and western blot assay was applied to detect surface markers CD9, CD81 and CD63. RESULTS AND CONCLUSION: Extraction of exosomes was completed by all the three methods, and round or oval membranous vesicles were observed under the transmission electron microscope. The protein content and purity of exosomes was highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group, and there were significant differences among the three groups (P < 0.05). Under the same protein concentration, surface specific markers, CD81, CD63 and CD9, were expressed highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group. The operating time was significantly lower in the Exobiology Quick kit group compared with the other two groups (P < 0.05). To conclude, despite a longer operating time, the differential ultracentrifugation method is a rational method to extract enough exosomes with relative high purity.
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Inositol,also called cyclohexanehexol,is a water-soluble Vitamin (Vitamin B8),which is widely distributed in plants and animals.It is an essential and indispensable nutrient for human and animals to maintain normal physiological functions with multiple bioactivities.Inositol can promote the maturity of a variety of ingredients in pulmonary surfactants,participating in the regulation of cell growth and surviving,biofilm formation,as well as the information transmission processes of a variety of signaling molecules.Therefore,inositol plays a key role in the growth and development of children and is closely related to pediatric developmental diseases.In order to provide a new way of thinking for prevention and treatment of inositol and its molecular mechanisms in pediatric developmental diseases,this article reviews the relationship between inositol and pediatric clinical diseases and its research progresses combined with the results of our laboratory studies.
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Lectins are the tools for the determination of sugar chain structure
Recently, lectin arrays have become a popular new technology; therefore, lectins with specific sugar-binding properties are required
The objective of the study was to isolate a novel lectin from Pleurotus ferulae mushrooms and characterize its various biological activities
A novel lectin was extracted with deionized water, precipitated from the aqueous extract using 75% saturated [Nfi] 2
The activity was tested using hemagglutination assays, and carbohydrate-binding specificity was determined by glycan microarray analysis
Its effects on the mitogenic activity of mouse splenocytes were determined by MTT assay. The novel lectin was adsorbed on ion-exchange chromatography DEAE-cellulose and shown as a band with the molecular mass of 17.5 kDa on a SDS-PAGE and as a single 35.0-kDa peak in gel filtration on Superdex G-75
The hemagglutinating activity of the lectin was inhibited by D-glucose, lactose, D-galactose, and galactosamine. The lectin was stable on 60°C
The hemagglutinating activity of lectin was reduced by 50% at 70°C. At 80°C, it was further reduced to 6.25% of its original activity. The hemagglutinating activity was the highest at pH 6-9. Moreover, its hemagglutinating activity was inhibited by Mg[2+] and Ca[2+] ions
The lectin isolated from P. ferulae in the current study possessed highly potent hemagglutinating and proliferative activities toward mouse splenocytes
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To explore novel antifatigue agents targeting with AMPA receptor, 10 compounds were synthesized and their structures were confirmed by 1H NMR, ESI-MS and elemental analysis. 1-BCP was treated as the leading compound. The antifatigue activities were evaluated by weight-loaded forced swimming test, and the AMPA receptor binding affinities were tested with radioligand receptor binding assays. The results unveiled that 5b appeared to possess potent antifatigue activities and high affinity with AMPA receptor, which deserved further studies.
Subject(s)
Animals , Benzamides , Chemistry , Pharmacology , Dioxoles , Chemistry , Pharmacology , Fatigue , Piperidines , Chemistry , Pharmacology , Radioligand Assay , Receptors, AMPA , Metabolism , SwimmingABSTRACT
BACKGROUND:A group of nuclease-like proteins were previously purified from Eisenia foetida tissues, exploring primary structures of these proteins wil help to uncover basic structure characteristics of them and provide foundations for the study addressing the relationship of their structures and functions. OBJECTIVE:To explore primary structures of nuclease-like proteins EWD1 and EWD2. METHODS:Edman degradation method was used to sequence the N-terminal amino acids of EWD1 and EWD2, acid hydrolisis method was used to analyze amino acid compositions of EWD1 and EWD2, LC-MS/MS was used to analyze some peptide sequences within the proteins, and MALDI-TOF-MS was used to calculate the number of the disulfide bonds and the contents of polysaccharides. RESULTS AND CONCLUSION:Among the amino acid compositions in EWD1 and EWD2, the sum contents of aspartate and asparagines were the highest (al nearly 10%), the contents of hydrophobic amino acids were also high, and the contents of cysteine was low. The EWD1 and EWD2 had similar amino acid compositions with other nucleases. Edman degradation results showed that, the N-terminal sequences of the large subunit of EWD1 were in turn as fol ows:D, E, W, V, Y, P;the N-terminal sequences of EWD2 were as fol ows:L, L, G, P, Y, K, P, K, C. The results of LC-MS/MS indicated the two proteins were novel proteins;MALDI-TOF-MS results showed that 8 cysteine residues formed 4 disulfide bonds in EWD1, 6 cysteine residues formed 3 disulfide bonds in EWD2. EWD1 and EWD2 were al glycoproteins, the content of polysaccharides was 17.3%in EWD1 and 15.6%in EWD2.
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<p><b>OBJECTIVE</b>To investigate the effects of naringin on the proliferation, differention and maturaion of rat calvarial osteoblasts (ROB).</p><p><b>METHOD</b>Segregated neonatal SD rat skull, enzyme digestion to obtain ROB. The culture medium was replaced every three days. Serial subcultivation proceeded when cells covered with 80% culture dish. Naringin supplemented into the culture at 1 x 10(-4), 1 x 10(-5), 1 x 10(-6), 1 x 10(-7) mol x L(-1) respectively. MTT method was adopted in proliferation analysis and the activity of ALP was examined after induced 9 days. Search the best concentration and supplemented into the medium, then the osteogenic differentiation markers including the secretion amount of osteocalcin, osteopontin and bone morphogenetic protein-2 were compared between the naringin-supplemented group and the control. Total RNA was isolated and the mRNA level of bFGF, IGF-1, Runx-2, Osterix, ERa and ERbeta was investigated by Real time RT-PCR. Total protein also was isolated and the expression ERa, ERbeta and collagen I was examined by Western blot. After the addition of ICI 182.780, an inhibitor of the estrogen signal pathway, these index also was examined and the changes were compared.</p><p><b>RESULT</b>The ROB proliferation was motivated by naringin dose-dependently. And it evidently leads to osteogenic process and maturation. 1 x 10(-5) mol x L(-1) is the best concentration. Naringin improved the secretion of osteocalcin, osteopontin, bone morphogenetic protein-2 and collagen I significantly. Besides, it can also enhanced the mRNA level of bFGF, IGF-1, Runx-2, Osterix, ERalpha and ERbeta. While all these effects can be restrained by ICI 182.780.</p><p><b>CONCLUSION</b>The naringin with final concentration of 1 x 10(-5) mol x L(-1) enhances the osteogenic differentiation and maturation of ROB significantly, while the promoting effects vanished after the addition of ICI 182.780. These results suggesting that naringin is one of the phytoestrogens and have the activity of bone formation may via estrogen signal pathway, it can be developed into a new drug for osteoporosis therapy.</p>
Subject(s)
Animals , Rats , Alkaline Phosphatase , Genetics , Metabolism , Cell Differentiation , Cell Proliferation , Cells, Cultured , Drugs, Chinese Herbal , Pharmacology , Flavanones , Pharmacology , Insulin-Like Growth Factor I , Genetics , Metabolism , Osteoblasts , Cell Biology , Metabolism , Osteocalcin , Genetics , Metabolism , Rats, Sprague-Dawley , Skull , Cell Biology , MetabolismABSTRACT
BACKGROUND:The viability of human umbilical cord-derived mesenchymal stem cel s is often declined with the commonly used transplantation storage solution in clinics, which may influence the therapeutic effects of cel ular transplantation. However, reasons for this are stil unknown. OBJECTIVE:To investigate the role of oxidative stress in the reduction of human umbilical cord-derived mesenchymal stem cel s viability in the storage process during clinical transplantation and to observe the effects of radical scavenger on the results. METHODS:Human umbilical cord-derived mesenchymal stem cel s were harvested and cultured in normal saline for 0, 2, 4 and 6 hours at room temperature. Intracel ular reactive oxygen levels were detected at those time points. Antioxidant enzyme activities and levels of malondialdehyde were measured to determine the intracel ular oxidative stress levels after storage. Cel adhesion rate changes were retested after adding N-acetyl cysteine to the storage solution. RESULTS AND CONCLUSION:The reactive oxygen levels in human umbilical cord-derived mesenchymal stem cel s were increased significantly after normal saline storage and levels of malondialdehyde were increased in a time-dependent manner. Activities of superoxide dismutase, catalase and glutathione peroxidase were al reduced. Addition of N-acetyl cysteine into the storage medium decreased the reactive oxygen levels and improved the human umbilical cord-derived mesenchymal stem cel s viabilities. Experimental findings indicate that, increased reactive oxygen species in human umbilical cord-derived mesenchymal stem cel s is one of the reasons for reduced cel viability. Adding the radical scavenger N-acetyl cysteine can improve the storage effects of human umbilical cord-derived mesenchymal stem cel s.
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BACKGROUND:Umbilical cord-derived mesenchymal stem cel s are gaining more attention in clinical treatments. Cel viability prior to transplantation has a direct impact on clinical prognosis. Despite trypan blue staining is a widely performed procedure to assess the viability of umbilical cord-derived mesenchymal stem cel s, it cannot reflect the functional capacity of those cel s accurately because of some subjective factors. OBJECTIVE:To explore sensitive and accurate assay for the functions of umbilical cord-derived mesenchymal stem cel s. METHODS:Human umbilical cord-derived mesenchymal stem cel s were isolated and cultured in vitro. Cultured umbilical cord-derived mesenchymal stem cel s were preserved in 0.9%saline for 0, 2, 4 and 6 hours at 4 ℃. Various methods (trypan blue staining, AnnexinV-PI, terminal deoxynucleotidyl transferase dutp nick end labeling, cel counting kit-8, live-dead assay, cel adherent assay) were used to determine the viability of post-storage umbilical cord-derived mesenchymal stem cel s, and the results were compared with colony-forming efficiency, a measure of cel function. RESULTS AND CONCLUSION:Human umbilical cord-derived mesenchymal stem cel s cultured in vitro showed a spindle shape and attached growth, the third-generation umbilical cord-derived mesenchymal stem cel s were positive for CD29, CD44, CD105, and negative for CD 34 and CD 45. Umbilical cord-derived mesenchymal stem cel s incubated in the adipogenic and osteogenic medium were both positive. Cel viability measured with trypan blue correlated moderately with colony-forming efficiency, while the percentage of viable cel s measured with other methods correlated better with colony-forming efficiency, among which adherent assay was the most obvious. It is proved that cel adherent assay-measured viability is the most accurate indicator.
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ObjectiveTo induce monocyte-derived dendritic cells(MoDC)from acute myeloid leukemia (AML) and healthy persons by rhCD40L in normal human AB serum system in vitro and to identify the morphology and phenotype of MoDC. MethodsPeripheral blood mononuclear cells(PBMNC)of AML and healthy persons were cultured in RPMI 1640 media including human AB serum, GM-CSF, rhIL-4 and rhCD40L, respectively. MoDC were identified by morphological features, surface antigen expression and the ability to stimulate T cells. ResultsAfter cultured for 7 days, MoDC displayed typical morphology with elongated dendritic process,and upregulation of the costimulatory molecules CD40,CD80,CD86 and CD83.The morphology and expression of costimulatory molecules were not significantly different between AML and healthy persons (P＞0.05),but were significantly different between rhCD40L group and without rhCD40L group (P＜0.05). MoDC had the ability to activate T cells, and there were no statistical differences between AML and healthy persons(P ＞0.05), but were significant differences between rhCD40L group and without rhCD40L group (P＜0.05). MoDC started to secrete IL-12 on day 5, and there was no statistical differences between AML and healthy persons(P＞0.05),and had differences between rhCD40L group and without rhCD40L group (P＜0.05).ConclusionMoDC can be cultured from the peripheral blood of AML and healthy persons.There were no significant differences in morphology and phenotype.Monocyted-derived DC can be used as an alternative to generate leukemia-specific cytotoxic T cells,especially in the presence of rhCD40L.
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<p><b>BACKGROUND</b>The signal transducer and activator of transcription 6 (STAT6) expression in lung epithelial cells plays a pivotal role in asthma pathogenesis. Activation of STAT6 expression results in T helper cell type 2 (Th2) cell differentiation leading to Th2-mediated IgE production, development of allergic airway inflammation and hyperreactivity. Therefore, antagonizing the expression and/or the function of STAT6 could be used as a mode of therapy for allergic airway inflammation.</p><p><b>METHODS</b>In this study, we synthesized a 20-mer phosphorothioate antisense oligonucleotide (ASODN) overlapping the translation starting site of STAT6 and constructed STAT6 antisense RNA (pANTI-STAT6), then transfected them into murine spleen lymphocytes and analyzed the effects of antagonizing STAT6 function in vitro and in a murine model of asthma.</p><p><b>RESULTS</b>In vitro, we showed suppression of STAT6 expression and interleukin (IL)-4 production of lymphocytes by STAT6 ASODN. This effect was more prominent when cells were cultured with pANTI-STAT6. In a murine model of asthma associated with allergic pulmonary inflammation in ovalbumin (OVA)-sensitized mice, local intranasal administration of fluorescein isothiocyanate (FITC)-labeled STAT6 ASODN to DNA uptake in lung cells was accompanied by a reduction of intracellular STAT6 expression. Such intrapulmonary blockade of STAT6 expression abrogated signs of lung inflammation, infiltration of eosinophils and Th2 cytokine production.</p><p><b>CONCLUSION</b>These data suggest a critical role of STAT6 in the pathogenesis of asthma and the use of local delivery of STAT6 ASODN as a novel approach for the treatment of allergic airway inflammation such as in asthma.</p>
Subject(s)
Animals , Female , Mice , Asthma , Drug Therapy , Metabolism , Blotting, Western , Cell Differentiation , Cells, Cultured , Interleukin-4 , Metabolism , Lymphocytes , Metabolism , Mice, Inbred C57BL , Oligonucleotides, Antisense , Chemistry , Pharmacology , Phosphates , Pharmacology , RNA, Antisense , Chemistry , Pharmacology , Reverse Transcriptase Polymerase Chain Reaction , STAT6 Transcription Factor , Genetics , Metabolism , Th2 Cells , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To optimize a pretreatment method of urine proteomics in children with primary nephrotic syndrome.</p><p><b>METHODS</b>Urine from children with primary nephrotic syndrome was treated in different pH and isolated by cold acetone precipitation for different durations. Then the amounts and kinds of proteins were compared by quantify, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE) in order to optimize a way to deal with urine protein.</p><p><b>RESULTS</b>Most proteins were obtained at pH 2.7. The amounts of protein precipitated by acetone for 0.5 hr was obviously less than those precipitated for 1 and 2 hrs (P<0.05), while there was no significant difference between the amount of protein precipitated for 1 and for 2 hrs. Protein precipitated by cold acetone for 1 hr at pH 2.7 was selected as the best pretreatment method. Satisfactory 2-DE maps can be acquired.</p><p><b>CONCLUSIONS</b>Urine protein can be best obtained at pH 2.7 and precipitated by cold acetone for 1 hr.</p>