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Aim To explore the effects of ethanol extraction of Dysosma versipellis on the proliferation and apoptosis of renal clear cell carcinoma OS-RC-2 cells and the underlying mechanism. Methods After treated with Dysosma versipellis, the proliferation of OS-RC-2 cells was detected by the CCK-8 assay and clone formation assay. The migration rate of cells was detected by thewound healing assay and Transwell assay. The level of ROS was detected by the reactive oxygen detection kit. The common targets between Dysosma versipellis and ROS were obtained by the network pharmacology. The above common targets were analyzed by KEGG. The apoptosis rate and cell cycle were detected by the flow cytometry, and the key proteins of MAPK signaling pathway and the levels of apoptosis related proteins were measured by Western blot. Results The results showed that Dysosma versipellis significantly inhibited the cell viability and migration ability of 0S-RC-2 cells, and up-regulated the level of ROS. Network pharmacology analysis showed a total of 165 common targets between Dysosma versipellis and ROS. KEGG analysis of the common targets revealed that there were significant changes in the MAPK signaling pathway. The results of Western blot showed that after treated with Dysosma versipellis, the protein level of JNK and the ratio of p-ERK/ERK were down-regulated. Besides, the protein level of caspase-9 and Bcl-2 declined, while the levels of cleaved caspase-9 and Bax were promoted. The flow cytometry results showed that Dysosma versipellis could significantly promote the apoptosis rate,down-regulate the cells in Gl-phase,while up-regulate the cells in G2/M-phase. The results of the rescue experiment showed that co-administration of NAC and Dysosma versipellis could significantly reverse the cell viability and apoptosis rate, the level of apop-totic related proteins, as well as the protein levels of MAPK pathway,when compared to treated with Dysosma versipellis alone in OS-RC-2. Conclusion In summary, Dysosma versipellis may inhibit the MAPK signaling pathway via the changes in ROS,further promoting apoptosis rate and decline cell proliferation in OS-RC-2 cell line.
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To investigate the antidepressant mechanism of Shenling Kaixin Granules(SLKX) in treating chronic unpredictable mild stress(CUMS) model rats. Ninety male SD rats were randomly divided into control group, model group, Shugan Jieyu Capsules(110 mg·kg~(-1)) group and SLKX low-(90 mg·kg~(-1)), medium-(180 mg·kg~(-1)), and high-dose(360 mg·kg~(-1)) groups. Depression rat model was replicated by CUMS method. After treatment, the behavioral changes of rats were evaluated by sugar preference, open field, elevated cross maze and forced swimming experiments. The contents of interleukin 1 beta(IL-1β), tumor necrosis factor α(TNF-α), brain-derived neurotrophic factor(BDNF) and 5-hydroxytryptamine(5-HT) in serum were determined by enzyme linked immunosorbent assay(ELISA), and the activities of superoxide dismutase(SOD) and catalase(CAT) in hippocampal CA1 region were also detected. Pathological changes in hippocampal CA1 region were detected by hematoxylin-eosin(HE) staining, and Western blot was used to determine the expression of nerve growth factor(NGF), BDNF, phospho-tyrosine kinase receptor(p-TrkB)/TrkB, phospho-cAMP-response element binding protein(p-CREB)/CREB, nuclear factor E2 related factor 2(Nrf2), heme oxygenase 1(HO-1), B-cell lymphoma-2(Bcl-2)/Bcl-2 associated X protein(Bax) and caspase-3 in hippocampal CA1 region. RESULTS:: showed that compared with the control group, the model group had decreased sugar preference, reduced number of entries and time spent in the center of open field and shortened total distance of movement, reduced number of entries and proportion of time spent in open arm, and increased number and time of immobility in forced swimming experiment. Additionally, the serum contents of IL-1β and TNF-α and the expression of caspase-3 were higher, while the contents of BDNF and 5-HT, the activities of SOD and CAT in hippocampal CA1 region, the expressions of NGF, BDNF, p-TrkB/TrkB, p-CREB/CREB, HO-1 and Bcl-2/Bax, and the Nrf2 nuclear translocation were lower in model group than in control group. Compared with the conditions in model group, the sugar preference, the number of entries and time spent in the center of open, total distance of movement, and the number of entries and proportion of time spent in open arm in treatment groups were increased while the number and time of immobility in forced swimming experiment were decreased; the serum contents of IL-1β and TNF-α and the expression of caspase-3 were down regulated, while the contents of BDNF and 5-HT, the activities of SOD and CAT in hippocampal CA1 region, the expressions of NGF, BDNF, p-TrkB/TrkB, p-CREB/CREB, HO-1, Bcl-2/Bax, and Nrf2 nuclear translocation were enhanced. In conclusion, SLKX might regulate the Nrf2 nucleus translocation by activating BDNF/TrkB/CREB pathway, lower oxidative stress damage in hippocampus, inhibit caspase-3 activity, and reduce apoptosis of hippocampal nerve cells, thereby playing an antidepressant role.
Subject(s)
Rats , Male , Animals , bcl-2-Associated X Protein/metabolism , Caspase 3/metabolism , Nerve Growth Factor/metabolism , Brain-Derived Neurotrophic Factor/metabolism , Signal Transduction , Tumor Necrosis Factor-alpha/metabolism , Serotonin/metabolism , NF-E2-Related Factor 2/metabolism , Rats, Sprague-Dawley , Antidepressive Agents/pharmacology , Hippocampus/metabolism , Superoxide Dismutase/metabolism , Sugars/pharmacology , Depression/genetics , Stress, Psychological/metabolismABSTRACT
This study aims to explore the neuroprotective mechanism of ginsenoside Re(GS-Re) on drosophila model of Parkinson's disease(PD) induced by rotenone(Rot). To be specific, Rot was used to induce PD in drosophilas. Then the drosophilas were grouped and respectively treated(GS-Re: 0.1, 0.4, 1.6 mmol·L~(-1); L-dopa: 80 μmol·L~(-1)). Life span and crawling ability of drosophilas were determined. The brain antioxidant activity [content of catalase(CAT), malondialdehyde(MDA), reactive oxygen species(ROS), superoxide dismutase(SOD)], dopamine(DA) content, and mitochondrial function [content of adenosine triphosphate(ATP), NADH:ubiquinone oxidoreductase subunit B8(NDUFB8) Ⅰ activity, succinate dehydrogenase complex, subunit B(SDHB) Ⅱ activity] were detected by enzyme-linked immunosorbent assay(ELISA). The number of DA neurons in the brains of drosophilas was measured with the immunofluorescence method. The levels of NDUFB8 Ⅰ, SDHB Ⅱ, cytochrome C(Cyt C), nuclear factor-E2-related factor 2(Nrf2), heme oxygenase-1(HO-1), B-cell lymphoma/leukemia 2(Bcl-2)/Bcl-2-assaciated X protein(Bax), and cleaved caspase-3/caspase-3 in the brain were detected by Western blot. The results showed that model group [475 μmol·L~(-1) Rot(IC_(50))] demonstrated significantly low survival rate, obvious dyskinesia, small number of neurons and low DA content in the brain, high ROS level and MDA content, low content of SOD and CAT, significantly low ATP content, NDUFB8 Ⅰ activity, and SDHB Ⅱ activity, significantly low expression of NDUFB8 Ⅰ, SDHB Ⅱ, and Bcl-2/Bax, large amount of Cyt C released from mitochondria to cytoplasm, low nuclear transfer of Nrf2, and significantly high expression of cleaved caspase-3/caspase-3 compared with the control group. GS-Re(0.1, 0.4, and 1.6 mmol·L~(-1)) significantly improved the survival rate of PD drosophilas, alleviated the dyskinesia, increased DA content, reduced the loss of DA neurons, ROS level, and MDA content in brain, improved content of SOD and CAT and antioxidant activity in brain, maintained mitochondrial homeostasis(significantly increased ATP content and activity of NDUFB8 Ⅰ and SDHB Ⅱ, significantly up-regulated expression of NDUFB8 Ⅰ, SDHB Ⅱ, and Bcl-2/Bax), significantly reduced the expression of Cyt C, increased the nuclear transfer of Nrf2, and down-regulated the expression of cleaved caspase-3/caspase-3. In conclusion, GS-Re can significantly relieve the Rot-induced cerebral neurotoxicity in drosophilas. The mechanism may be that GS-Re activates Keap1-Nrf2-ARE signaling pathway by maintaining mitochondrial homeostasis, improves antioxidant capacity of brain neurons, then inhibits mitochondria-mediated caspase-3 signaling pathway, and the apoptosis of neuronal cells, thereby exerting the neuroprotective effect.
Subject(s)
Animals , Reactive Oxygen Species/metabolism , Antioxidants/pharmacology , Oxidative Stress , NF-E2-Related Factor 2/metabolism , Caspase 3/metabolism , Parkinson Disease/genetics , bcl-2-Associated X Protein/metabolism , Neuroprotective Agents/pharmacology , Kelch-Like ECH-Associated Protein 1/metabolism , Drosophila/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Apoptosis , Superoxide Dismutase/metabolism , Adenosine Triphosphate/pharmacologyABSTRACT
OBJECTIVE: To provide the basis for biapenem against Acinetobacter baumannii infection in ICU patients in our hospital. METHODS: One hundred-five strains of ABA were collected. The MICs of biapenem against bacteria were measured by double broth dilution method. Four therapeutics regimens (0.3 g, q12 h; 0.3 g, q8 h; 0.3 g, q6 h; 0.6 g, q12 h) in traditional short-time infusion, extended infusion(3, 4h) and two-step infusion(1.5-5.5 h) were simulated by using the Monte Carlo simulation, then the PTAs and CRFs were calculated. RESULTS: CFRs of all treatments of biapenem against ABA infection in ICU subjects were 90% except biapenem 0.3g, q12 h(0.5 h) and 0.3 g, q12 h (two-step infusion for 1.5h). PTAs of the whole regimens were >90% when MIC≥2 mg·L-1. Extended infusion and two-step infusion got the similar PTAs and CFRs with a similar time when the regimens obtained target, more than short infusion obviously. CONCLUSION: Biapenem alone should avoid for ABA infection as expiry therapy in ICU population in our hospital. Extended infusion and two-step infusion should be priority selection as target therapy according to sensitivity test.
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<p><b>OBJECTIVE</b>To investigate the expressions of aquaporins (AQPs) in the mouse prostatic tissue and their significance, and to provide some evidence for a deeper insight into the physiological function and regulation of AQP expressions in normal and diseased prostatic tissues.</p><p><b>METHODS</b>The mRNA expressions of AQP0 - 4 in the mouse prostatic tissue were determined by RT-PCR, and the expressions and localizations of AQP1 and AQP3 proteins were characterized by Western blot and immunohistochemistry.</p><p><b>RESULTS</b>RT-PCR exhibited the mRNA expressions of AQP1 and AQP3, but not those of AQP0, AQP2 and AQP4 in the prostate tissue, while Western blot showed the expression of the AQP1 protein with the relative molecular mass (RMM) of 28 000 and those of the glycosylated and non-glycosylated AQP3 proteins with the RMM of 35 000 and 27 000, respectively. Immunohistochemistry indicated the strong expression of AQP1 in the cyst and plasma membrane of the secretary cells and that of AQP3 in the stroma cells of the prostate.</p><p><b>CONCLUSION</b>The AQP1 and AQP3 genes were expressed in the secretary epithelia of the mouse prostate tissue, which suggests that AQP1 and AQP3 may play an important role in the secretion of prostatic fluid.</p>
Subject(s)
Animals , Male , Mice , Aquaporin 1 , Genetics , Metabolism , Aquaporin 3 , Genetics , Metabolism , Mice, Inbred BALB C , Prostate , Metabolism , PathologyABSTRACT
<p><b>AIM</b>The piezoelectric properties and cytotoxicity of a porous lead-free piezoelectric ceramic for use as a direct bone substitute were investigated.</p><p><b>METHODOLOGY</b>Cold isostatic pressing (CIP) was applied to fabricate porous lithium sodium potassium niobate (Li0.06Na0.5K0.44) NbO3 specimens using a pore-forming method. The morphologies of the CIP-processed specimens were characterized and compared to those of specimens made by from conventional pressing procedures. The effects of the ceramic on the attachment and proliferation of osteoblasts isolated from the cranium of 1-day-old Sprague-Dawley rats were examined by a scanning electron microscopy (SEM) and methylthiazol tetrazolium (MTT) assay.</p><p><b>RESULTS</b>The results showed that CIP enhanced piezoelectricity and biological performance of the niobate specimen, and also promoted an extracellular matrix-like topography of it. In vitro studies showed that the CIP-enhanced material had positive effects on the attachment and proliferation of osteoblasts.</p><p><b>CONCLUSION</b>Niobate ceramic generated by CIP shows a promise for being a piezoelectric composite bone substitute.</p>
Subject(s)
Animals , Rats , Biocompatible Materials , Chemistry , Toxicity , Bone Substitutes , Toxicity , Cell Adhesion , Cell Proliferation , Cells, Cultured , Ceramics , Toxicity , Coloring Agents , Electrochemistry , Materials Testing , Microscopy, Electron, Scanning , Niobium , Toxicity , Osteoblasts , Oxides , Toxicity , Porosity , Potassium , Toxicity , Pressure , Rats, Sprague-Dawley , Skull , Cell Biology , Stress, Mechanical , Surface Properties , Tetrazolium Salts , ThiazolesABSTRACT
Objective To study the rate of mother-to-child transmission (MTCT) on HIV-1. Methods All local residents from 8 townships in a region were screened for mothers who had a history of only one blood transfusion and 63 were found HIV-1 positive. A further study on these HIV-1 positive mothers and their children was conducted with the emphasis on the date of receiving blood transfusion, date and type of nationality, history regarding breastfeeding and so on. Sera specimens from 84 children born from 63 HIV-1 positive mothers were screened, using ELISA for HIV-1 antibody, and positive specimens were confirmed by Western-blot. Results The rate of MTCT was 32.1% (27/84) for children with all risk factors related to MTCT. Another 36.8% (7/19) were related to factors on intrauterine, intrapartum and breastfeeding, 35.7% (5/14) to intrapartum and breastfeeding factors, 14.3% (2/14) to intrauterine and intrapartum factors, 37.9% (11/29) to breastfeeding factor alone. By group combination analysis, the MTCT rate was 36.9% (24/65) with breastfeeding, 11.8% (2/17) with artificial feeding, and the former was significantly higher than the latter. Conclusion HIV-1 MTCT rate among mothers caused by a single blood transfusion varied with different risk factors. Breastfeeding played an important role in MTCT, appeared in our study.
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<p><b>OBJECTIVE</b>Using the functional connectivity analysis based on the underlying neurophysiological characteristic that epileptic discharges can induce change of brain default mode, to develop a technique for epileptogenic localization using functional MRI (fMRI) without simultaneous electroencephalography (EEG).</p><p><b>METHODS</b>A data-driven method that jointly employed independent component analysis and functional connectivity analysis was used for the resting functional MRI data analysis of 12 focal epileptic patients. The independent components were ranged according to the coefficients of the negative correlation between independent component time course and the signal temporal course in the region of posterior cingulate cortex. The results were comparatively studied with simultaneous EEG-fMRI.</p><p><b>RESULTS</b>In the 10 successful results from 12 patients underwent EEG-fMRI examination, the outcomes of eight subjects were concordant with pathological foci. While the results of all 10 patients processed by data-driven method were concordant with pathological foci, besides the other patients who failed to perform EEG-fMRI examination. Meanwhile, the default mode was well mapped in all patients.</p><p><b>CONCLUSIONS</b>The default mode-based functional connectivity analysis can localize the epileptogenic foci effectively without simultaneous EEG, besides to detect the default mode of epileptic patients.</p>
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Male , Epilepsy , Diagnosis , Magnetic Resonance Imaging , MethodsABSTRACT
Objective The aim of this study was to investigate the radiation dose caused by 18 F-fluorodeoxyglucose (FDG) in PET/CT examination and to optimize the concerned radiation protection. Methods Thirty patients from our conventional PET/CT examination were simple randomly selected, and they all underwent whole body PET/CT imaging. The radioactive dose of injected 18F-FDG was recorded. The internal radiation dose was calculated and the external radiation dose from patients was measured with the 451P-DE-SI ion chamber survey meter. The staff's dose was recorded with thermoluminescent detector (TLD). All dosimetry data were processed and analyzed statistically with Excel 2003. Results The injected radioactive dose of 18F-FDG was (432.9±51.8) MBq, and effective dose equivalent received per patient was (8.23±0.99) roSy. The correlation coefficient (r) of the dose equivalent rate and distance was-0.994 by power function curve fitting, and that of dose equivalent rate and time was - 0.988 by exponential curve fitting. The staff's dose was lower than the annual dose limit. Conclusions The patient's internal radiation dose caused by 18F-FDG in PET/CT examination is low, nonetheless, the clinician should always consider optimizing and minimizing the necessary radiation received by the patients. The patients having been injected with 18F-FDG should stay in one place to decrease their radiation to the public. From the medical point of view in optimizing radiation exposure, there may still be a potential to lower the injected 18F-FDG activity.
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<p><b>OBJECTIVE</b>To detect the serum autoantibodies against the cardiac beta(1)-adrenergic receptor and observe the clinical characteristics and response to carvedilol use in patients with chronic heart failure (CHF).</p><p><b>METHODS</b>Cardiac function was examined by echocardiography and levels of autoantibodies against cardiac beta(1)-adrenergic receptor were detected in 65 patients with CHF by means of enzyme linked immune assay. Carvedilol was added on ACEI, diuretics and digitalis regimen for a target dose of 50 mg/d. All patients were followed up for 6 months.</p><p><b>RESULTS</b>Autoantibodies against cardiac beta(1)-adrenergic receptor were detected in 30 patients (group 1) and not detected in the remaining 35 patients (group 2). The achieved target dose of carvedilol was significantly higher in group 1 than that in group 2 [(36.25 +/- 14.31) mg/d vs. (25.97 +/- 8.83) mg/d, P < 0.01]. Heart rate was significantly higher in group 1 compared to group 2 [(94.19 +/- 14.46) beats/min vs. (86.56 +/- 15.88) beats/min, P < 0.05] before treatment and heart rate and blood pressure of both groups decreased significantly (P < 0.01) and there was no significant difference between two group (P > 0.05) after 6 months treatment. LVEDD and LVESD were significantly larger while LVEF significantly lower in group 1 patients than those in group 2 patients (all P < 0.05) before treatment and LVEDD and LVESD decreased and LVEF increased significantly in both groups (all P < 0.01 vs. before treatment) and there was on significant difference in LVEDD, LVESD and LVEF between two groups (all P > 0.05) post 6 months treatment. Moreover, average titer of autoantibodies against the cardiac beta(1)-adrenergic receptors significantly decreased after 6 months treatment (1:119.35 vs. 1:72.21, P < 0.01).</p><p><b>CONCLUSION</b>The detection of autoantibodies against the cardiac beta(1)-adrenergic receptors is related to severer cardiac dysfunction and autoantibodies title decrease was found with improved cardiac function after standard therapy (ACEI, digitalis, betablocker) in patients with CHF.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Adrenergic beta-Antagonists , Therapeutic Uses , Autoantibodies , Blood , Carbazoles , Therapeutic Uses , Follow-Up Studies , Heart Failure , Blood , Allergy and Immunology , Propanolamines , Therapeutic Uses , Receptors, Adrenergic, beta-1 , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To confirm the expression and distribution of aquaporin 5 (AQP5) in rat nasal mucosa of experimental allergic rhinitis and to investigate the relationship between AQP5 and allergic rhinitis.</p><p><b>METHODS</b>Twenty-four healthy Wistar rats both male and female weighting 200-300 g were divided into two groups randomly, one was testing group (n = 12), the other was comparing group (n = 12). Generally sensitized rats in testing group were given repeated local booster sensitization into nasal cavity. Twelve normal rat nasal mucosa and twelve nasal mucosa from rat with allergic rhinitis were used. The distribution of AQP5 in normal rat nasal mucosa and nasal mucosa in rat with allergic rhinitis were observed by immunofluorescence technique. Furthermore, the expression of AQP5 in normal rat nasal mucosa and nasal mucosa from rat with allergic rhinitis were studied by immunohistochemical staining.</p><p><b>RESULTS</b>Hematoxylin and eosin staining showed that there was obvious inflammation reaction, a large quantity of glands hyperplasia and acidophils soaked in nasal mucosa from rat with allergic rhinitis. (2) Both immunofluorescence technique and immunohistochemical staining showed that the distribution of AQP5 in normal rat nasal mucosa was in accordance with that in nasal mucosa from rat with allergic rhinitis on the whole. AQP5 expressed mainly in the membrane and cytoplasm of the epithelium in the glands, ducts and cilia. (3) The statistical analysis of the immunohistochemical staining showed that the quantity of AQP5 in rat nasal mucosa with allergic rhinitis (156.37 +/- 1.93) was obviously higher than that in the normal rat nasal mucosa (178.52 +/- 1.94). There was statistical significance between the two groups (t = 28.08, P < 0.01).</p><p><b>CONCLUSIONS</b>The hypersecretion of glands has a close relationship with the high expression of AQP5 in allergic rhinitis.</p>
Subject(s)
Animals , Female , Male , Rats , Aquaporin 5 , Metabolism , Nasal Mucosa , Metabolism , Rats, Wistar , Rhinitis, Allergic, Perennial , MetabolismABSTRACT
The cDNA fragments of interest were amplified using Sj lambda ZipLox library as the templates by PCR and then cloned into a eukaryotic expression vector p-CMV-GH; A small number of DNA fragments inserted in the recombinants was identified by restriction cleavage, EST sequencing and bioinformatical analysis; mice were injected intramuscularly with the expression library (L-CMV-SjR) or sublibraries(L-CMV-SjR1, L-CMV-SjR2 and L-CMV-SjR3), immunized mice were challenged with Schistosoma japonicum cercariae on day 35, the levels of IgG antibodies in sera from the immunized mice were detected by ELISA. The results demonstrated that a partial cDNA expression library of S.j, with approximately 10(5) transformants, was constructed, most of the recombinants contained the insert DNA fragments of interest, and these fragments had the features of protein-coding sequences for Schistosome. There were no significant differences for the levels of IgG antibodies in sera from all of the immunized groups. Mice immunized with L-CMV-SjR, L-CMV-SjR1 and L-CMV-SjR2 developed significant protective effect against Sj infection compared to control mice injected with the empty plasmid, the rate of worm reduction was about 30%.
Subject(s)
Animals , Mice , Antibodies, Helminth , Blood , Allergy and Immunology , Antigens, Helminth , Genetics , Allergy and Immunology , DNA, Complementary , Disease Models, Animal , Gene Expression , Gene Library , Schistosoma japonicum , Genetics , Allergy and Immunology , Schistosomiasis japonica , Vaccination , Methods , Vaccines, DNA , Genetics , Allergy and ImmunologyABSTRACT
Objective To observe the brain activation of interictal epiletiform discharges(IEDs) and to localize the epileptogenic foci of epilepsy.Methods The electroencephalogram(EEG)and functional MRI data of 12 focal epileptic patients were acquired using a combination of EEG and functional MRI simultaneously.The IEDs onset time detected with EEG were set as the time parameters in an event- related paradigm of functional MRI analysis.The spatial and temporal characters of IEDs activation were analyzed in detail.In order to confirm the consistency of this method,all patients were scanned repeatedly and the results were correlated with clinical evaluation.Results Of the 12 patients,valid data from EEG- fMRI were obtained from 10 patients in a total of 18 sessions..Compared with the structural foci,the epileptic foci localization results of eleven sessions were good,five sessions were fairly good,and two sessions were poor.The results obtained from six patients in two separate sessions were concordant.respectively.Moreover,thalamic activation was detected in ten sessions,cerebellar activation was detected in all sessions,and the deactivation was found in the default mode loci in nine sessions. Conclusion The method of performing EEG and fMRI simultaneously can potentially be a useful tool in epilepsy research.