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OBJECTIVE@#To explore the anti-inflammatory effects of ethyl lithospermate in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine-derived macrophages and zebrafish, and its underlying mechanisms.@*METHODS@#3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide (MTT) assays were performed to investigate the toxicity of ethyl lithospermate at different concentrations (12.5-100 µ mol/L) in RAW 264.7 cells. The cells were stimulated with LPS (100 ng/mL) for 12 h to establish an inflammation model in vitro, the production of pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor α (TNF-α) were assessed by enzyme linked immunosorbent assay (ELISA). Western blot was used to ascertain the protein expressions of signal transducer and activator of transcription 3 (STAT3), nuclear factor kappa B (NF-κB) p65, phospho-STAT3 (p-STAT3, Tyr705), inhibitor of NF-κB (IκB) α, and phospho-I κB α (p-IκB α, Ser32), and confocal imaging was used to identify the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705). Additionally, the yolk sacs of zebrafish (3 days post fertilization) were injected with 2 nL LPS (0.5 mg/mL) to induce an inflammation model in vivo. Survival analysis, hematoxylin-eosin (HE) staining, observation of neutrophil migration, and quantitative real-time polymerase chain reaction (qRT-PCR) were used to further study the anti-inflammatory effects of ethyl lithospermate and its probable mechanisms in vivo.@*RESULTS@#The non-toxic concentrations of ethyl lithospermate have been found to range from 12.5 to 100 µ mol/L. Ethyl lithospermate inhibited the release of IL-6 and TNF-α(P<0.05 or P<0.01), decreased IκBα degradation and phosphorylation (P<0.05) as well as the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705) in LPS-induced RAW 264.7 cells (P<0.01). Ethyl lithospermate also decreased inflammatory cells infiltration and neutrophil migration while increasing the survival rate of LPS-stimulated zebrafish (P<0.05 or P<0.01). In addition, ethyl lithospermate also inhibited the mRNA expression levels of of IL-6, TNF-α, IκBα, STAT3, and NF-κB in LPS-stimulated zebrafish (P<0.01).@*CONCLUSION@#Ethyl lithospermate exerts anti-Inflammatory effected by inhibiting the NF-κB and STAT3 signal pathways in RAW 264.7 macrophages and zebrafish.
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Animaux , Souris , Facteur de transcription NF-kappa B/métabolisme , Lipopolysaccharides , Danio zébré , Inhibiteur alpha de NF-KappaB/métabolisme , Interleukine-6/métabolisme , Facteur de nécrose tumorale alpha/métabolisme , Facteur de transcription STAT-3/métabolisme , Inflammation/métabolisme , Anti-inflammatoires/usage thérapeutiqueRésumé
Objective: To evaluate the effectiveness and safety of ultrasound-guided percutaneous cannulation for extracorporeal membrane oxygenation (ECMO) in children. Methods: In this retrospective observational study, 66 cases who underwent non-cardiac surgery ECMO in pediatric intensive care unit (PICU) of Shanghai Children's Hospital from May 2016 to April 2021 were collected. The demographics, model of ECMO support, type and size of arteriovenous cannulas, way of catheterization and complications were recorded and summarized. Patients were divided into percutaneous cannulation group and surgical cannulation group according to catheterization strategies. The demographics, duration of cannulation and ECMO support, ECMO weaning rate and hospital survival rate were compared among two groups. χ2 and nonparametric rank sum test were used for comparison. Results: Among the 66 patients who received ECMO, 38 were male and 28 were female, with age 44.5 (12.0, 83.5) months and weight 15.0 (10.0, 25.0) kg; 21 patients underwent percutaneous cannulation, with a success rate of 95% (20 cases). Point-of-care ultrasound was performed for all percutaneous cannulation cases. The duration of percutaneous cannulation was significantly shorter than that of surgical cannulation (26.0 (23.3, 30.3) vs. 57.0 (53.8, 64.0) min, Z=6.31, P<0.001). Successful percutaneous cannulation cases were aged 70.5 (23.8, 109.5) months, and their weight was 23.2 (13.6, 37.0) kg. Ten cases were initially given veno-venous (VV) ECMO support, and 10 cases were given veno-arterial (VA) ECMO support. ECMO arterial cannulas were sized from 8 F to 17 F, and venous cannulas sized from 10 F to 19 F. For VV-ECMO, the right internal jugular and femoral veins were used as vascular access, while VA-ECMO used right internal jugular vein-femoral artery or right femoral vein-left femoral artery approach. Only one patient suffered severe complication (superior vena cava perforation). There was no catheter-related bloodstream infection. Conclusion: Ultrasound-guided percutaneous cannulation for ECMO can be performed with a high rate of success and safety in children.
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Adulte , Enfant , Femelle , Humains , Mâle , Cathétérisme , Chine , Oxygénation extracorporelle sur oxygénateur à membrane , Études rétrospectives , Échographie interventionnelle , Veine cave supérieureRésumé
BACKGROUND@#The new emerging avian influenza A H7N9 virus, causing severe human infection with a mortality rate of around 41%. This study aims to provide a novel treatment option for the prevention and control of H7N9.@*METHODS@#H7 hemagglutinin (HA)-specific B cells were isolated from peripheral blood plasma cells of the patients previously infected by H7N9 in Jiangsu Province, China. The human monoclonal antibodies (mAbs) were generated by amplification and cloning of these HA-specific B cells. First, all human mAbs were screened for binding activity by enzyme-linked immunosorbent assay. Then, those mAbs, exhibiting potent affinity to recognize H7 HAs were further evaluated by hemagglutination-inhibiting (HAI) and microneutralization in vitro assays. Finally, the lead mAb candidate was selected and tested against the lethal challenge of the H7N9 virus using murine models.@*RESULTS@#The mAb 6-137 was able to recognize a panel of H7 HAs with high affinity but not HA of other subtypes, including H1N1 and H3N2. The mAb 6-137 can efficiently inhibit the HA activity in the inactivated H7N9 virus and neutralize 100 tissue culture infectious dose 50 (TCID50) of H7N9 virus (influenza A/Nanjing/1/2013) in vitro, with neutralizing activity as low as 78 ng/mL. In addition, the mAb 6-137 protected the mice against the lethal challenge of H7N9 prophylactically and therapeutically.@*CONCLUSION@#The mAb 6-137 could be an effective antibody as a prophylactic or therapeutic biological treatment for the H7N9 exposure or infection.
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Animaux , Humains , Souris , Anticorps monoclonaux/usage thérapeutique , Anticorps neutralisants/usage thérapeutique , Anticorps antiviraux , Hémagglutinines , Sous-type H1N1 du virus de la grippe A , Sous-type H3N2 du virus de la grippe A , Sous-type H7N9 du virus de la grippe A , Vaccins antigrippaux , Grippe chez les oiseaux , Grippe humaine/prévention et contrôleRésumé
The present study investigated the effect of Modified Dihuang Decoction in improving ovarian reserve in mice through the Bcl-2-related mitochondrial apoptosis pathway. Forty-eight adult female BALB/c mice were randomly divided into the following six groups with eight mice in each group: a blank group, a model group, a femoston group(three cycles of treatment with 0.13 mg·kg~(-1) estradiol tablets for 2 days and 1.43 mg·kg~(-1) estradiol and dydrogesterone tablets for 3 days), and high(64.74 g·kg~(-1))-, medium(43.16 g·kg~(-1))-, and low-dose(21.58 g·kg~(-1)) Modified Dihuang Decoction groups. Mice in other groups except the blank group received a single intraperitoneal injection of 12 mg·kg~(-1) cyclophosphamide and 1.2 mg·kg~(-1) busulfan to induce a model of diminished ovarian reserve(DOR), while those in the blank group received an equal volume of normal saline. Mice were treated with corresponding drugs for 15 d from the 36 th day, once per day, and the mice in the blank group and the model group were treated with an equal volume of normal saline. The general condition and oestrous cycle were observed. The serum hormone levels were detected with the enzyme-linked immunosorbent assay(ELISA). The morphological changes of ovaries were observed by HE staining. Western blot was used to detect the protein expression of cysteinyl aspartate specific proteinase-9(caspase-9), cleaved caspase-3, Bcl-2 associated X protein(Bax), Bcl-2, superoxide dismutase-2(SOD-2), and glutathione peroxidase-1(GPx-1). The mRNA expression of Bax and Bcl-2 was detected by real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR). The results showed that compared with the blank group, the model group showed body weight loss, disordered oestrous cycle, elevated serum levels of follicle-stimulating hormone(FSH) and luteinizing hormone(LH), reduced serum levels of estradiol(E_2), anti-mullerian hormone(AMH), and inhibin B(INHB), the declining number of ovarian follicles and granulosa layers, increased number of atretic follicles, up-regulated protein expression of caspase-9, cleaved caspase-3, and Bax and Bax mRNA expression in ovaries, and down-regulated protein expression of Bcl-2, SOD-2 and GPx-1, and Bcl-2 mRNA expression. Compared with the model group, the Modified Dihuang Decoction groups displayed restored body weight and oestrous cycle, decreased serum levels of FSH and LH, elevated serum levels of E_2, AMH, and INHB, increased number of ovarian follicles, thickened granulosa layers, and declining number of atretic follicles. Additionally, the protein expression of caspase-9, cleaved caspase-3, and Bax, and Bax mRNA expression was down-regulated, and the protein expression of Bcl-2, SOD-2, and GPx-1, and Bcl-2 mRNA expression was up-regulated. The results suggest that Modified Dihuang Decoction can regulate endocrine hormone, promote follicle growth and improve ovarian reserve by enhancing ovarian anti-oxidant capacity, inhibiting the Bcl-2-related mitochondrial apoptosis pathway, and further inhibiting cell apoptosis.
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Animaux , Femelle , Souris , Apoptose , Souris de lignée BALB C , Follicule ovarique , Réserve ovarienne , OvaireRésumé
Lipopolysaccharide (LPS)-induced inflammation causes massive threatening diseases, such as sepsis, acute lung injury and multiple organ dysfunction syndrome. Efficient treatment to prevent inflammation is crucial in LPS-induced inflammatory diseases. Heat-clearing Chinese medicines (CMs) have been used to ameliorate LPS-induced inflammation in China for centuries. Heat-clearing CMs regulate inflammatory pathways, thereby inhibiting the release of inflammatory factors. This review aimed to introduce promising heat-clearing CMs countering LPS-induced inflammation in the last 5 years, exploring the underlying molecular mechanisms.
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Objective To prepare human foreskin tissue acellular matrix graft(AMGs), and explore the toxicity of the extracts on rabbit urethral mucosal cells. Methods After preparing the decellularized matrix of human foreskin tissue, the characteristics of decellularized matrix were observed by Masson staining, and the pore size was observed by scanning electron microscope. Results Masson staining showed that the cells were removed, and the scaffold was composed of collagen. Scanning electron microscopy showed that the pore size of the scaffold was about 20-50 μm, which was uniformly distributed. The appropriate concentration of AMGs matrix extract had no obvious toxicity to urethral mucosal cells. Conclusion The acellular matrix of human foreskin tissue is an ideal AMGs scaffold material. The prepared AMGs material has low cytotoxicity and has practical value.
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Using the lipidomics method based on UHPLC-Q-Exactive Orbitrap/MS, the change of phospholipid metabolism in lung tissue of mice induced by lipopolysaccharide (LPS)-induced acute lung injury was analyzed to observe the regulation of abnormal lipids by Jiegeng Decoction and to explore the regulation effect of Jiegeng Decoction on LPS-induced acute lung injury. The lung tissue samples from control group, model group, dexamethasone (positive drug) group, and Jiegeng Decoction group were collected and the lipid components of the sample were extracted. All procedures over mice were performed in accordance with the Guidelines for Care and Use of Laboratory Animals of Nanjing University of Chinese Medicine, and the experiments were approved by the Animal Ethics Committee of our university. The lipidomics technique of UHPLC-Q-Exactive Orbitrap/MS was used to study change of phospholipids in lung tissue of each group. LPS induced acute lung injury in mice with metabolic abnormalities of phospholipids, the specific performance of the PC was significantly upregulated, phosphatidyl ethanolamine (PE), phosphatidyl glycerol (PG), phosphatidyl serine (PS),phosphatidylinositol (PI) and other metabolic disorders, Jiegeng Decoction have a certain role in these phospholipids. LPS-induced acute lung injury caused disturbances of phospholipid in vivo, and Jiegeng Decoction regulates metabolic phospholipids.
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A quantitative analytical method for multi-components with a single-marker (QAMS) was established for simultaneous determination of neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction. The separation was performed on a Waters CORTECS T3 column (2.1 mm × 100 mm, 2.7 μm), with the mobile phase consisting of 0.05% phosphate acid solution-acetonitrile for gradient elution. The column temperature was 30 ℃, and flow rate was 0.5 mL·min-1. Using chlorogenic acid as an internal reference, the relative correlation factors of neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid were calculated following UHPLC, as 0.928 0, 0.546 2, 1.099 8, 0.872 1, 1.086 8, 0.739 2, 1.056 6, respectively. The results were compared with those obtained by the external standard method to verify the feasibility, rationality and repeatability of QAMS method. There was no significant difference in assay results between QAMS and the external standard method. In conclusion, the QAMS method is accurate and feasible, and could be used to determine the content such as neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction.
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OBJECTIVE@#To study the effects of miR-221 on the biological activity of childhood acute lymphoblastic leukemia cells and its mechanism.@*METHODS@#Bone marrow mononuclear cells (BMNC) were isolated from bone marrow samples of ALL children diagnosed in our hospital from May 2018 to November 2018. The cells were divided into control group, miR-221-NC group and miR-221 group. After transfection according to the instructions of Lipofectamine 2000 kit, the levels of miR-221 in each group were detected by RT-PCR. Flow cytometry was used to detect the effects of miR-221 on cell cycle and apoptosis. Transwell assay was used to detect cell migration and invasion. Western blot was used to detect the effects of miR-221 on proliferating cell nuclear antigen (PCNA), Caspase 3, Cyclin D1 and MMP-9 proteins in BMNC. Luciferase reporter gene assay was used to detect the targeting relationship between miR-221 and Wnt gene.@*RESULT@#The expression level of miR-221 in the miR-221 group was significantly higher than that in the control group and the miR-221-NC group (P<0.05). MTT assay showed that, after transfection for 2, 3, 4 and 5 days, the cell proliferation level in miR-221 group was significantly lower than that in the control group and the miR-221-NC group (P<0.05). The cell ratio of G/G phase was (73.25±8.1)% in the miR-221 group, which was significantly higher than that in the control group and the miR-221-NC group (P<0.05); moreover, the cell ratio of S phase in the miR-221 group was (12.37±1.6)%,which was significantly lower than that in the control group and the miR-221-NC group (P<0.05). The percentage of apoptotic cells in the miR-221 group was (24.68±3.87)%, which was significantly higher than that in the control group and the miR-221-NC group (P<0.05). Transwell cell invasion experiment showed that the number of invasive cells in the miR-221 group was 23.42±3.62, which was significantly lower than that in the control group and the miR-221-NC group (P<0.05). Transwell cell migration assay showed that the number of migrating cells in the miR-221 group was 34.86±5.32, which was significantly lower than that in the control group and the miR-221-NC group (P<0.05). The relative level of PCNA, Cyclin D1 and MMP-9 in the miR-221 group was 0.26±0.03, 0.17±3.61 and 0.14±0.02, respectively, which was significantly lower than those in the control group and the miR-221-NC group (P<0.05), while the relative level of Caspase-3 in the miR-221 group was 0.37±0.05, which was significantly higher than that in the control group and the miR-221-NC group (P<0.05). Luciferase reporter assay showed that the activity of luciferase in Wnt wild type plasmid was significantly inhibited by miR-221 (P<0.05).@*CONCLUSION@#miR-221 can inhibit the proliferation, migration and invasion of BMNC, moreover can promote cell apoptosis, which may be related with the inhibition of Wnt/β- catenin signaling pathway.
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Enfant , Humains , Caténines , Lignée cellulaire tumorale , Prolifération cellulaire , microARN , Leucémie-lymphome lymphoblastique à précurseurs B et T , Voie de signalisation WntRésumé
Objective: To investigate the molecular mechanism and potential active components of Jiegeng Decotion (JD) with the antitussive and expectorant effects. Methods: Target proteins related with phlegm and cough were selected through mining literature and retrieving in DrugBank and TTD database, and the main active components and potential target proteins from JD were computed and analyzed by DOVIS 2.0 and Cytoscape 3.0 to build a molecular-protein regulatory network. Results: A total of 38 target proteins and 472 small molecules were initially screened based on the pathological mechanism which is related with phlegm and cough. Molecular docking results showed that 78 molecules (five from Platycodi Radix and 73 from Glycyrrhizae Radix et Rhizoma, and their structural characteristics analysis were in accordance with the “rules of generic drugs”) were found in JD with higher docking score (Score ≥ 7) of target protein. According to the results of molecular docking, 128 molecular-target protein data pairs with high docking scores (Score ≥ 7) were selected, and then 26 major active components of JD (saponins and flavonoids, etc.) and 13 target proteins were identified by using Network analyzer. Conclusion: The active components of JD could regulate over-inflammatory response on the respiratory tract, improve the lung function, inhibit the over-expression of mucin, and reduce the reaction of the stimulation on cough center through acting on the main target proteins (TLR4, MMP9, IKK2, etc), thereby achieving the antitussive and expectorant effects.
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Objective The mechanism of luteal phase defect remains unclear. To investigate the mechanism of BuShen ZhuYun Decoction on the gonadotropin secretion in the pituitary gland, we observed the effects of medicated serum of BuShen ZhuYun Decotion on the secretion of gonadotropin-follicle-stimulating hormone (FSH) and luteotropic hormone (LH) in rat pituitary cells.Methods The BuShen ZhuYun Decotion was administered to the female SD rats by gavage to prepare the serum containing BuShen ZhuYun Decoction. The CCK-8 method was used to detect the effect of cetrorelix acetate powder for injection, medicated serum and gonadotropin releasing hormone (GnRH) on cell activity. In the maximum non-toxic concentration, we used cetrorelix acetate powder for injection to block the GnRH receptor (GnRHR) in pituitary cells and established the GnRHR antagonistic model. Then we treat the model group with medicated serum (model group). Moreover, we established the blank group (normal pituitary cells), the cetrorelix group (intervented with cetrorelix for 6 hours), and medicated serum group (intervented with medicated serum for 24 hours). 20nmol/L GnRH was used to stimulate cells for 6h. The contents of FSH and LH in the supernatant of each group and the mRNA expression of FSHβ, LHβ and GnRHR were detected.Results Compared with that of the blank group, the supernatant levels of FSH and LH in the Cetrorelix group decreased significantly \[(3.91±0.36) mIU/mL vs (2.26±0.22) mIU/mL, (8.94±0.57) mIU/mL vs (3.35±0.59) mIU/mL, P<0.05)\]. In contrast, the levels of LH significantly increased \[(8.94±0.57) mIU/mL vs (10.79±0.60) mIU/mL, P<0.05)\]; Compared with the cetrorelix group, the levels of FSH and LH in both medicated serum group and model group increased significantly (P<0.05). Compared with the blank group, the mRNA level of FSH and LH in the cetrorelix group decreased significantly \[(0.95±0.23) mIU/mL vs (0.58±0.12) mIU/mL, (0.98±0.14) mIU/mL vs (0.27±0.21) mIU/mL, P<0.01) \], and the mRNA expression of GnRHR increased in the cetrorelix group \[(0.97±0.13) mIU/mL vs (1.77±0.26) mIU/mL, P<0.01) \]; The mRNA levels of FSH and LH in the medicated serum group were increased (P<0.05). Compared with the cetrorelix group, the mRNA expression of FSHβ mRNA and LHβ were both increased in the medicated serum group and model group (P<0.05), the mRNA expression of GnRHR decreased (P<0.01).Conclusion It is suggested that the therapeutic mechanism of BuShen ZhuYun Decotion may be related to the improvement of GnRH receptor expression.
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Two new phenolic glycosides, 7S, 8R-urolignoside-9'-O-β-D-glucoside (1) and scrophenoside G (2), were isolated and identified from the seeds of Ginkgo biloba L., a famous traditional medicine and functional food around the world. Their structures were elucidated by spectroscopic methods (1D and 2D NMR, HR-ESI-MS, and CD), and the comparisons of spectroscopic data with the reported values in the literature.
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Ginkgo biloba , Chimie , Hétérosides , Chimie , Structure moléculaire , Phénols , Chimie , Extraits de plantes , Chimie , Plantes médicinales , Chimie , Graines , Chimie , Analyse spectraleRésumé
Two new phenolic glycosides, 7S, 8R-urolignoside-9'-O-β-D-glucoside (1) and scrophenoside G (2), were isolated and identified from the seeds of Ginkgo biloba L., a famous traditional medicine and functional food around the world. Their structures were elucidated by spectroscopic methods (1D and 2D NMR, HR-ESI-MS, and CD), and the comparisons of spectroscopic data with the reported values in the literature.
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Ginkgo biloba , Chimie , Hétérosides , Chimie , Structure moléculaire , Phénols , Chimie , Extraits de plantes , Chimie , Plantes médicinales , Chimie , Graines , Chimie , Analyse spectraleRésumé
Objective To establish a QAMS method for content determination of six compositions (chlorogenic acid, caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and loganin) from Lonicerae Japonicae Caulis; To verify the feasibility and applicability of this method in quality control of Lonicerae Japonicae Caulis. Methods Chlorogenic acid was set as internal reference substance. The HPLC analysis was performed on a Waters Symmetry C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase consisted of acetonitrile and 0.4% phosphoric acid solution in gradient elution manner at a flow rate of 1 mL/min. The column temperature was maintained at 35 ℃, and the detection wavelength was set at 327 nm for chlorogenic acid, caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and 236 nm for loganin. Results The relative correction factors of caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and loganin were established; there was no obvious difference between calculated value of QAMS and measured value of external standard method. Conclusion The quality control mode of QAMS can be used for multi-index synchronization quality evaluation of the six compositions from Lonicerae Japonicae Caulis.
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A new naphthalene derivative and three known compounds were isolated from the petroleum ether extract of the bulbs of Eleutherine americana by using various chromatographic techniques, such as column chromatography over silica gel and semi-preparative HPLC. Their structures were elucidated by spectroscopic date (MS, UV, IR, NMR), which were identified as eleutherol B (1), 4,8-dihydroxy-3-methoxy-1-methylanthraquinone-2-carboxylic acid methyl ester (2), 8-hydroxy-3,4-dimethoxy-1-methylanthraquinone-2-carboxylic acid methyl ester (3), and isoeleutherine (4). Compound 1 is a new compound. The diastolic blood vessels activity of compound 1 and 2 were potent, reaching 82.5% and 85.3% at the concentration of 10 μmol·L⁻¹, which were basically the same as that of the positive drug tanshinone ⅡA.
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The changes of endogenous metabolites in urine samples that come from pneumonia patients of 6 months to 6 years old children were analyzed by metabolomics methods based on gas chromatography and mass spectrometry (GC-MS).The aim of this study was to analyze and study the pathogenesis of endogenous metabolites in children with pneumonia and the pathogenesis of pneumonia susceptibility.The urine samples were collected and divided into normal children group (NC group,n=29),first infection with pneumonia group (FIP group,n=35),and repeated infection with pneumonia group (RIP group,n=31).The urine metabolic profile of pneumonia was obtained by GC-MS.Principal component analysis(PCA) and partial least squares-discriminant analysis (PLS-DA) were used to analyze the data.The results were analyzed by one-way analysis of variance and Fold change.Finally,there was significant difference between the normal group and the pneumonia group,the significant metabolites were serine,histidine,proline,norleucine,glutamine,stearic acid,valine,isoleucine with p value<0.05 and Fold change>5,and indole-3-acetic acid,creatine,ethanolamine,mannosylglycerol and fructose were significant between the two pneumonia groups with p value<0.05.The urinary metabolites demonstrated that amino acid metabolism and glucose metabolism were the main metabolic pathways and responsible for the susceptibility to pneumonia.
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Exhaled breath condensate (EBC) was analyzed by gas chromatography-mass spectrometry (GC-MS / MS) in childhood asthma and healthy control, aiming to find the potential markers of EBC in children with asthma, and provide a scientific reference for its pathogenesis and early screening. EBC samples were collected from 21 asthmatic children (age (8. 2 ±1. 6) years) and 17 healthy children ( age (8. 1 ±1. 3) years). GC-MS / MS was used to obtain the full scan data of chemical components. Cluster analysis was performed on the two groups of metabolites by principal component analysis (PCA), and potential biomarkers were found using Metaboanalyst 3. 0 attributable metabolic pathways. The results showed that the EBC metabolic maps of asthmatic group and normal group were very different, and eight endogenous potential biomarkers were identified, suggesting that starch and sucrose metabolism, lysine degradation, aminoglycan nucleoside metabolism, phenylalanine metabolism may play important roles in the development of asthma in children.
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<p><b>OBJECTIVE</b>To investigate the inhibitory effect of giganteaside D (GD) on hepatocellular carcinoma and its molecular mechanisms.</p><p><b>METHODS</b>The inhibitory effects of GD on Hep 3b cells were determined using MTT assay and colony formation assay. The morphological changes of Hep 3b cells after GD treatment were observed by electron microscopy, and the cell cycle changes was analyzed using flow cytometry. The cell apoptosis and mitochondrial potential collapse in the treated cells were tested with Hoechst staining assay and flow cytometry. The expression levels of Bcl-2, PARP and key proteins in MAPK pathway were detected using Western blotting.</p><p><b>RESULTS</b>GD showed a significant inhibitory effect on Hep 3b cells with an ICvalue of 16.08 µmol/L at 72 h. Flow cytometric analysis demonstrated that the phases of cell cycle remained unchanged and a sub-G1 peak (from 3.3% to 33.6%) appeared as GD concentration increased. GD-induced apoptosis was further conformed by Hoechst staining assay, and flow cytometry showed increased mitochondrial potential collapse in the cells. Western blotting demonstrated the cleavage of PARP, decrease of Bcl-2 and p-Erk1/2 (Thr202/Tyr204), and activation of p-p38 (Thr180/Tyr182) and p-JNK (Thr183/Tyr185) in GD-treated cells.</p><p><b>CONCLUSIONS</b>GD has significant inhibitory effect against hepatocellular carcinoma cells in vitro by inducing apoptosis possibly in association with the MAPK signaling pathway.</p>
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BACKGROUND:The angiogenesis may be related to the proliferation of neural stem ceils,but there is still no unified view.OBJECTIVE:To observe the influence of angiogenesis on neural stem cell proliferation in the subventricular zone of rats after focal cerebral ischemia/reperfusion.METHODS:Male Sprague-Dawley rats were randomly divided into normal group,sham group,vascular endothelial growth factor (VEGF)+cerebral ischemia/reperfusion group,normal saline (NS)+cerebral ischemia/reperfusion group.The injection was done via the lateral cerebral ventricle.Then,each group was subdivided into four groups (1,2,7,14 days after ischemia/reperfusion).Focal cerebral ischemia/reperfusion models were made by the thread method.After modeling,the corresponding intervention was given in each group.The expression changes of Nestin and vWF mRNA in the subventricular zone were detected in all groups by immunohistochemical staining and real-time PCR,respectively.RESULTS AND CONCLUSION:There was a certain increase in vWF and Nestin positive expression in the subventricular zone after cerebral ischemia/reperfusion.At 7 days after ischemia,the expression of vWF mRNA and Nestin reached the peak,indicating the proliferation of neural stem cells in the subventricular zone after cerebral ischemia/reperfusion is associated with the time of angiogenesis.In addition,the expression of vWF mRNA and Nestin was significantly higher in the VEGF+cerebral ischemia/reperfusion group than the other two groups,indicating angiogenesis could promote the proliferation of neural stem ceils in the subventricular zone of rats after cerebral ischemia/reperfusion.
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Objective To investigate the clinical detection of bladder vascular resistance index in female overactive bladder patients. Methods Bladder vascular resistance index in 40 female overactive bladder patients and 20 healthy controls was measured by transabdominal pulsed wave spectral Doppler imaging. Results Significant difference between female overactive bladder patients and healthy women controls was observed in the resistive index(0.81 ± 0.06 vs.0.64 ± 0.06,P<0.05). Conclusions The results indicate that bladder ischemia may play a role in female overactive bladder patients.