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Objective·To analyze the expression changes of adhesion G protein-coupled receptor F1(ADGRF1)in the occurrence and development of pancreatic ductal adenocarcinoma(PDAC),and explore the impact of ADGRF1 on the proliferation of PDAC cells and the potential molecular mechanisms that promote PDAC progression.Methods·The expression of ADGRF1 at mRNA level was analyzed based on the Gene Expression Omnibus(GEO)database and The Cancer Genome Atlas(TCGA)database,respectively.The expression of ADGRF1 in normal pancreatic ductal epithelial cells(hTERT-HPNE)and various PDAC tumor cells was detected by using real-time fluorescence quantitative PCR(qPCR)and Western blotting.Immunohistochemical staining(IHC)was used to detect the differential expression of ADGRF1 in cancer tissues and adjacent tissues of PDAC patients.After knocking down ADGRF1 with small interfering RNA(siRNA)transfection,the changes in the proliferation ability of PDAC AsPC-1 and SW1990 cells were detected through CCK8 assay and plate cloning experiment.Stable overexpression of ADGRF1 was constructed in PDAC Patu8988 cell line,and the proliferation changes induced by overexpression of ADGRF1 were evaluated through CCK8 assay.RNA sequencing(RNA-seq),gene set enrichment analysis(GSEA),and immune infiltration analysis were utilized to predict signaling pathways associated with ADGRF1-mediated promotion of PDAC cancer progression.Results·Analysis of the TCGA database and GEO database revealed higher expression of ADGRF1 mRNA in PDAC tissues compared to normal pancreatic tissues(all P=0.000).qPCR and Western blotting results demonstrated up-regulation of ADGRF1 mRNA and protein levels in various PDAC cells compared to hTERT-HPNE cells(all P<0.05).IHC results confirmed higher ADGRF1 expression in PDAC cancer tissues compared to adjacent tissues.Furthermore,downregulation of ADGRF1 inhibited the proliferation of PDAC AsPC-1 and SW1990 cell lines,while overexpression of ADGRF1 promoted the proliferation of Patu8988 cells(all P<0.05).RNA-seq,GSEA enrichment analysis,and immune infiltration analysis revealed that ADGRF1 expression was related to signaling pathways such as interferon-α(IFN-α),tumor necrosis factor-α(TNF-α),and nuclear factor κB(NF-κB).Conclusion·ADGRF1 is highly expressed in PDAC cells and tissues,and promotes the proliferation of PDAC cells via immune-related signaling pathways.
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Acute kidney injury (AKI) is an important factor for the occurrence and development of CKD. The protective effect of dihydroartemisinin on AKI and and reported mechanism have not been reported. In this study, we used two animal models including ischemia-reperfusion and UUO, as well as a high-glucose-stimulated HK-2 cell model, to evaluate the protective effect of dihydroartemisinin on premature senescence of renal tubular epithelial cells in vitro and in vivo. We demonstrated that dihydroartemisinin improved renal aging and renal injury by activating autophagy. In addition, we found that co-treatment with chloroquine, an autophagy inhibitor, abolished the anti-renal aging effect of dihydroartemisinin in vitro. These findings suggested that activation of autophagy/elimination of senescent cell might be a useful strategy to prevent AKI/UUO induced renal tubular senescence and fibrosis.
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Animais , Rim , Injúria Renal Aguda/induzido quimicamente , Isquemia , Traumatismo por Reperfusão/tratamento farmacológico , Autofagia , ReperfusãoRESUMO
Objective@#: This study aimed to investigate the changes and significance of microRNA155 levels in serum of patients with cerebral small vessel disease (CSVD). @*Methods@#: Thirty patients with CSVD who met the inclusion criteria were selected and divided into eight patients with lacunar infarction (LI) group and 22 patients with multiple lacunar infarction (MLI) combined with white matter lesions (WML) group according to the results of head magnetic resonance imaging (MRI). Thirty samples from healthy volunteers without abnormalities after head MRI examination were selected as the control group. The levels of serum microRNA155 in each group were determined by real-time polymerase chain reaction, and the correlation between microRNA155 in the serum of patients with CSVD and the increase of imaging lesions was analyzed by Spearman correlation analysis. @*Results@#: Compared with the control group, the serum microRNA155 level in the LI group, MLI combined with WML group increased, the difference was statistically significant (p<0.05); serum microRNA155 level was positively correlated with the increase of imaging lesions (p<0.05). @*Conclusion@#: The change of serum microRNA155 level in patients with CSVD may be one of its self-protection mechanisms, and the intensity of this self-protection mechanism is positively correlated with the number of CSVD lesions.
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Objective: To explore the feasibility of quantitative measurement of cerebral iron using brain network atlas based on quantitative susceptibility mapping (QSM). Methods QSM images of 43 right-handed healthy adult volunteers were registered, which were smoothed and mapped to the standard brain using Matlab software. ROI of bilateral globus pallidus, putamen, caudate nucleus, hippocampus, thalamus, frontal cortex, parietal cortex and occipital cortex were selected from Brainnetome Atlas. The magnetic susceptibility was measured using Matlab software, and ROI of the above areas were manually sketched and measured. The correlation of the magnetic susceptibility values measured with the above 2 methods and the iron concentration results obtained from brain tissue staining of autopsy were analyzed, and the correlation between the magnetic susceptibility values and age were analyzed. Results: The highest measurement value of brain magnetic susceptibility values from Matlab and manually drawn ROI were all found in globus pallidus, then in the putamen, and the lowest was in hippocampus. The brain magnetic susceptibility values measured with Matlab and manual ROI were all highly consistent with autopsy results (r=0.920, P=0.003; r=0.856, P=0.014). The magnetic susceptibility values of male at frontal cortex measured from Matlab ROI was higher than that of female (P0.05), nor between left and right hemispheres brain regions measured with 2 methods (all P>0.05). Conclusion: Quantitative measurement of cerebral iron based on QSM imaging and Brainnetome Atlas has high accuracy and feasibility. The content of brain iron tends to increase with aging. The magnetic susceptibility values of frontal cortex have sex differences.
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Objective:To evaluate the effect of propofol combined with sevoflurane anesthesia on GABA A receptor (GABA AR)α 1/α 2 subunit homeostasis in hippocampus of rats with mild cognitive impairment (MCI). Methods:Healthy clean-grade male Sprague-Dawley rats, aged 16-18 months, weighing 450-550 g, were anesthetized.MCI was induced by ligation of bilateral common carotid arteries after anesthesia.Morris water maze test was used to select the rats with MCI at 30 days after establishment of the model.The rats with MCI were divided into 4 groups ( n=18 each) using a random number table method: sham operation group (group Sham), sevoflurane anesthesia group (group S), propofol anesthesia group (group P), and propofol combined with sevoflurane anesthesia group (group SP). The rats inhaled 3% sevoflurane for 3 h in group S. Propofol 40 mg·kg -1·h -1 was intravenously infused for 3 h in group P. The rats inhaled 1.7% sevoflurane, and propofol 20 mg·kg -1·h -1 was intravenously infused for 3 h simultaneously in group SP.Open reduction and internal fixation was performed after tibial fracture was induced in S, P and SP groups.Y-maze test was performed at 14 days after operation to assess cognitive function.The expression of potassium-chloride cotransporter-2 (KCC2), sodium-potassium-chloride cotransporter 1 (NKCC1), GABA ARα 1 and GABA ARα 2 was determined using Western blot. Results:Compared with group Sham, the percentage of time of staying at novel arm was significantly decreased, the expression of hippocampal KCC2 and GABA ARα 1 was down-regulated, and the expression of NKCC1 and GABA ARα 2 was up-regulated in S and P groups ( P<0.05), and no significant change was found in the parameters mentioned above in group SP ( P>0.05). Compared with group S or group P, the percentage of time of staying at novel arm was significantly increased, the expression of hippocampal KCC2 and GABA ARα 1was up-regulated, and the expression of NKCC1 and GABA ARα 2 was down-regulated in group SP ( P<0.05). Conclusion:The mechanism by which propofol combined with sevoflurane anesthesia does not aggravate the postoperative cognitive dysfunction may be related to maintaining GABA ARα 1/α 2 subunit homeostasis in rats with MCI.
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Iron is an important trace element in human body, and too much or too little iron can lead to corresponding pathological changes. Although there is no direct evidence that the destruction of iron homeostasis leads to neurodegenerative diseases, it is undeniable that the abnormal iron content is involved in the pathogenesis of most diseases. Having high resolution and sensitivity, quantitative susceptibility mapping (QSM) imaging is able to detect abnormal iron depositions in the early stage of most neurodegenerative diseases, therefore it is used as the main method to quantitatively measure iron content in vivo brain tissue in the study of neurodegenerative diseases in recent years. QSM imaging research progresses of iron metabolism in Alzheimer disease, Parkinson disease, Huntington disease, multiple sclerosis and amyotrophic lateral sclerosis were reviewed in this article.
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Hemifacial spasm (HFS) is a common cranial nerve disease,which seriously affects the beauty,visual function and quality of life of patients.The pathogenesis of the disease is still unclear and no effective drug therapy is found.Surgical treatment is the only way now.In this paper,the progress of etiology and pathophysiology,three dimensional imaging evaluation,electrophysiological evaluation,endoscopic application and microvascular decompression of HFS are discussed.
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Objective@#To evaluate the performance of Vitek MS, an automated mass spectrometry microbial identification system, in identification of clinical filamentous fungi isolates, which provide a new insight into filamentous fungi identification strategy.@*Methods@#Twenty-five isolates of filamentous fungi genetically confirmed by molecular sequencing method were inoculated in different culture conditions to analyze the potential influences of culture conditions on the identification of filamentous fungi with Vitek MS. Further evaluation of the performance of Vitek MS in filamentous fungi identification was carried out with 207 clinical isolates of filamentous fungi and the comparison of the results with those of morphology and sequencing analysis.@*Results@#The performance of Vitek MS in filamentous fungi identification was not significantly influenced by culture conditions. Vitek MS successfully identified 87.9% (182/207) of all tested filamentous fungi. Lacking of reference MS spectra in Vitek database was the main reason that the other isolates could not be identified. Vitek MS was superior to the traditional morphological method in identification of filamentous fungi especially non-Aspergillus molds at species level.@*Conclusion@#Vitek MS is an efficient approach for clinical filamentous fungi identification. Continuous enrichment of the species coverage in database will be of great importance for improving the performance of Vitek MS in identification of filamentous fungi.
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Objective To evaluate the application values of diffusion tensor tractography (DTT) combined with neuronavigation in microsurgery of insular gliomas. Methods The clinical data of 27 patients with insular gliomas, admitted to our hospital from March 2013 to October 2017, were analyzed retrospectively. All DTT images were transferred to the neuronavigation system, and the three-dimensional location of tumors and pyramidal tracts were re-constructed. Surgical approaches were designed and excision scopes were defined before the surgery. Techniques on how to distinguish and protect the key blood vessels and pyramidal tracts were discussed. The treatment efficacies were analyzed. Results Total lesion resection was achieved in 22 patients (81.5%), subtotal resection in 4 (14.8% ), and partial resection in one (3.7% ). Postoperative pathology indicated 7 were oligodendrogliomas, and 20 were astrocytomas, including WHO grade I in one, grade II in 18, and grade III in one. One patient had transient aphasia (recovery after two weeks), 2 experienced worsened hemiplegia on opposite side of their bodies (normal after one month), and the left 24 patients remained intact function after operation. Conclusions The combination of DTT and neuronavigation is safe and effective in surgical treatment for insular gliomas, which can protect the brain function at greatest degree and maximize lesion resection, and improve the postoperative quality of life.
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<p><b>OBJECTIVE</b>To develop a GC-FID method for the determination of borneol concentration in rat plasma and to investigate the pharmacokinetics after injection of novel-Xingnaojing.</p><p><b>METHOD</b>Novel-Xingnaojing was injected via by caudal vein injection. The blood samples were collected by posterior orbital venous plexus approach at 0.5, 1, 3, 5, 8, 12, 20, 30, 45 min. The drug in plasma was extracted with ethyl acetate and then detected by GC-FID, octadecane was used as the internal standard. The pharmacokinetic parameters were calculated by the software of Kinetica.</p><p><b>RESULT</b>The calibration curve was good linear in the range of 1.67-16.67 mg x L(-1). The extraction recoveries of low, medium and high concentration were (92.81 +/- 1.11)%, (85.38 +/- 0.86)% and (84.58 +/- 0.58)%, respectivley. And the RSDs of within-day and between-day were below 3.00%. Plasma concentration of borneol was consistent with the two-compartment open model. The pharmacokinetic parameters were that the t1/2alpha was (1.18 +/- 0.20) min, the t1/2beta was (22.27 +/- 6.85) min, the C(max)(Calc) was (18.76 +/- 2.10) mg x L(-1), the MRT was (23.84 +/- 7.67) min(-1), and the AUC was (100.00 +/- 15.85) mg x min x L(-1).</p><p><b>CONCLUSION</b>The GC-FID method developed can be applied to determination and pharmacokinetics. The borneol in novel-Xingnaojing is distributed and metabolized fast after being administrated.</p>
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Animais , Masculino , Ratos , Canfanos , Farmacocinética , Medicamentos de Ervas Chinesas , Farmacocinética , Ionização de Chama , Métodos , Ratos Sprague-DawleyRESUMO
<p><b>OBJECTIVE</b>To study in vitro release of Xingnaojing microemulsion and to investigate the release mechanism.</p><p><b>METHOD</b>The concentration of jasminoidin was determined by HPLC and the concentration of Aipian was determined by GC. In vitro release characteristics were conducted by dialysis technique. Model fitting was used to determine the kinetics and mechanism.</p><p><b>RESULT</b>Jasminoidin released completely within 2 h, fitting the Weibell model best. The release of borneol fitted first order model.</p><p><b>CONCLUSION</b>The release mechanisms of different types of medicines are quite different. The different types of medicines dissolve in the different phases in the microemulsion.</p>
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Cromatografia Líquida de Alta Pressão , Métodos , Medicamentos de Ervas Chinesas , Química , Emulsões , Peso MolecularRESUMO
BACKGROUND: Cell cycle has proved that about 90% stem cells were in resting state in the G0 phase; therefore, serum deprivation method which did not contain any growth factors and related nutrition additive was beneficial for separating glioma stem cells.OBJECTIVE: To explore the method of serum deprivation to culture glioma cell lines U87, and to isolate and identify the cancer stem cells which are in the U87 cell lines. METHODS: U87 cells were cultured in the medium containing both DMEM and L-glutamine for six days; cancer stem cells would be screened out; then the medium was changed for the neural stem cell culturing medium. The formation of tumor spheres and their differentiation characteristics were observed when they were inoculated onto serum-containing medium. The immunofluorescence staining of cells was employed to identify the surviving cells cultured in serum deprivation medium, tumor spheres and differentiated cells. RESULTS AND CONCLUSION: The serum deprivation method was used to select tumor stem cells successfully which expressed CD133 and could form tumor spheres. The tumor spheres had an ability of multi-differentiations, and the daughter cells expressed glial fibrillary acidic protein and neuron specific enolase. The U87 cell lines exist glioma stem cells which have the capacities of self-renewal and multi-differentiation.
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Objective To identify the accurate measurement of glomerular filtration rate (GFR) in chronic glomerulonephritis (CGN)patients. Methods Forty-two patients were enrolled in the study, including 15 females with age from 18 to 73 years old (mean 46 years old) and 27 males with age from 20 to 77 years old (mean 48 years old). The methods used for measuring GFR were classical dual plasma sample clearance method (tGFR), considered to be the gold standard,renal dynamic imaging method (dGFR), 24-hour creatinine clearance method (24hCcr). The difference and correlation amony them were analyzed. When the difference was significant, Pearson correlation and linear regression analysis were further performed. The difference of GFR detected by dGFR between left and right kidney of patients was compared simultaneously. A two-sided P value<0.05 was considered as statistically significant. Results Either dGFR or 24hCcr was statistically different from tGFR, but had excellent correlation with tGFR, and the coefficient was 0.916 (P=0.000) and 0.957 (P=0.000) respectively. The linear regressions correlation existed and the regression equations were tGFR=0.936 dGFR-4.648 (F=208.941, P=0.000), tGFR =0.887 24hCcr+2.919 (F=376.513, P=0.000) respectively. Difference had not statistically significance between left and right kidney of patients (P=0.591). Conclusions Neither dGFR nor 24hCcr can substitute tGFR, but both can reflect the GFR of the CGN patients safely and effectively. The decrease of GFR is homochronous for left and right kidney of CGN patients. Therefore, the 24hCcr can be chosen to evaluate the GFR in the hospitals without SPECT.