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Objective To explore the effect and potential mechanisms of melatonin combined with gemcitabine on the chemosensitivity of human pancreatic cancer cell line PANC-1.Methods Human pancreatic cancer cell line PANC-1 was trea-ted with gemcitabine alone or in combination with melatonin.Cell viability was assessed using CCK-8.Effect of melatonin and gem-citabine alone or in combination on the clonogenic capacity of PANC-1 cells were observed through colony formation experiments.Scratch assays and transwell experiments were conducted to evaluate cell migration ability.Reactive oxygen species(ROS)and mitochondrial membrane point JC-1 assay kit were used to determine reactive oxygen species synthesis and membrane potential levels.Intracellular Fe2+level was measured using ferrous ion fluorescent probe.The protein expression levels of LC3,P62,GPX4 and SLC7A11 in different treatment groups were detected by immunofluorescence and Western blotting.Results CCK-8 results showed that the viability of PANC-1 cells was inhibited by gemcitabine alone after 48 h and 72 h of treatment in a time-and dose-dependent manner.The cell viability of gemcitabine combined with melatonin group was significantly lower than that of gemcitabine group,and the cell viability decreased with the increase of melatonin concentration.Scratch assays,transwell experiments,and plate colony formation assay results demonstrated that the proliferation and migration of cells in the gemcitabine combined with the me-latonin group were significantly inhibited compared with the gemcitabine group.The levels of reactive oxygen species and Fe2+in PANC-1 in gemcitabine combined with the melatonin group were higher than those in the gemcitabine group,and the mitochondri-al membrane potential was significantly decreased(P<0.01).Western blotting and immunofluorescence results showed that the ra-tio of autophagy-related protein LC3-Ⅱ/LC3-Ⅰ in gemcitabine combined with the melatonin group was lower than that in the gem-citabine group,and the expression of P62 was up-regulated,and the expression of anti-iron death-related protein GPX4 and SLC7A11 was significantly inhibited(P<0.05),suggesting that melatonin combined with gemcitabine can inhibit autophagy and promote ferroptosis in PANC-1 cells.Conclusion Melatonin enhances the chemosensitivity of pancreatic cancer cell PANC-1 to gemcitabine by inhibiting autophagy and promoting ferroptosis of tumor cells.
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Objective:To explore the correlation between cognitive impairment and intestinal mucosal barrier injury in rats after chronic cerebral hypoperfusion(CCH), and to quantitatively analyze the changes in cognitive behavior of experimental rats caused by chronic cerebral hypoperfusion, as well as the expression changes of the intestinal mucosal barrier claudin-1 and osteopontin.Methods:Thirty male SD rats were randomly divided into CCH group ( n=15) and sham operation (SHAM) control group ( n=15). The CCH model was established by permanent ligation of bilateral common carotid arteries.Rats in the SHAM group only separated the common carotid artery without ligation.Four weeks later, open field experiment, object discrimination experiment, and Morris water maze experiment were used to detect the emotional arousal ability, the ability to explore new things, and the ability of spatial learning and memory in rats.HE staining and immunofluorescence experiments were conducted to detect the damage of rat ileum tissue.Western blot was used to detect OPN expression, and ELISA was used to detect serum OPN.SPSS 23.0 and GraphPad 8.0 statistical softwares were used to process the data, and the t-test and repeated measures one-way analysis of variance were used for data analysis. Results:In the open field test, compared with the SHAM group ((28.70±10.70)times, (1 030.45±81.51)cm), the number of standing and total exercise distance of rats in the CCH group ((16.70±7.13)times, (736.64±136.71)cm) were decreased( t=1.59, 4.16, both P<0.05). In the object discrimination experiment, the discrimination index of rats in the CCH group (0.44±0.26) was lower than that of the SHAM group (0.91±0.07, t=-7.76, P<0.05). Morris water maze positioning navigation experiment showed that the group main effect and time main effect were both significant( F=383.36, 153.87, P<0.05). Simple effect analysis showed that, compared with the SHAM group, the escape latency and total swimming distance of rats in CCH group increased( P<0.05). Space exploration experiment showed that, compared with SHAM group ((7.20±1.81)times, (9.96±2.95)s), the number of crossings of rats in CCH group ((3.00±0.82)times) decreased, and the incubation period ((29.70±6.28)s) was prolonged( t=4.65, 7.04, both P<0.05). The intestinal mucosal pathology score of SHAM group ((1.98±0.34)points) was lower than that of the CCH group ((4.52±0.27)points), and the difference was significant( t=18.53, P<0.01). Immunofluorescence experiment showed that, compared with SHAM group (125 028.58±33 077.39), the cumulative optical density of claudin-1 between the intestinal epithelial cells of the CCH group(47 154.50±7 507.29) decreased( t=16.10, P<0.01). Western blot experiment showed that, compared with the SHAM group (0.38±0.11), the expression of OPN in the intestines of the CCH group (1.20±0.95) increased( P<0.05). ELISA experiment showed that, compared with the SHAM group ((3.42±0.66)μg/L), the serum OPN content of the CCH group ((14.92±1.45)μg/L) significantly increased( P<0.05). The degree of cognitive impairment was negatively correlated with intestinal mucosal epithelial claudin-1 expression and serum OPN content( P<0.01). Intestinal mucosal epithelial claudin-1 expression was negatively correlated with serum OPN content ( r=-0.952, P<0.01). Conclusion:CCH may cause obvious cognitive impairment in rats and the destruction of the intestinal mucosal barrier.Serum OPN may be a potential serological marker of CCH-induced cognitive impairment and intestinal mucosal barrier destruction in rats.
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Objective:To investigate the correlation between sarcopenia and post-stroke cognitive impairment(PSCI)in elderly patients with first-time acute minor ischemic stroke.Methods:This was a prospective study.Elderly patients over 60 years of age with first-time acute minor ischemic stroke admitted to the Department of Neurology of the General Hospital of Western Theater Command from October 2018 to June 2019 were continuously enrolled.Patients received the SARC-F score assessment within 24h after admission and were divided into two groups according to their SARC-F scores: the non-sarcopenia group(SARC-F score<4)and the sarcopenia group(SARC-F score≥4). Cognitive function was assessed by using the Mini-Mental State Examination(MMSE)within 24 h of admission and at 3-month follow-up.Results:A total of 211 patients were enrolled in this study, including 31 patients(31/211, 14.69%)in the sarcopenia group and 180 patients(180/211, 85.31%)in the non-sarcopenia group.The incidence of PSCI was higher in the sarcopenia group than in the non-sarcopenia group(83.87% or 26/31 vs.55.56% or 100/180, χ2=8.814, P=0.003). The total MMSE score, orientation, immediate memory, attention, calculation and language functions were lower in the sarcopenia group compared with non-sarcopenia group( P<0.05). Logistic regression analysis showed that sarcopenia was an independent risk factor for PSCI( OR=3.478, 95% CI: 1.039-11.642, P=0.043)in the elderly with first-time acute minor ischemic stroke. Conclusions:Sarcopenia is an independent risk factor for PSCI in elderly patients with first-time acute minor ischemic stroke.Sarcopenia assessment in the acute phase of stroke might help doctors to assess the risk of PSCI and reduce the incidence of PSCI in stroke patients.
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@#Objective To analyze the relationship of cerebral infarction(CI) between VitB12,folic acid(FA) and homocysteine(HCY).Methods Patients with CI,aged between 50 to 80 years old were collected from our hospital in recent three years.Patients with benign paroxysmalpositional vertigo(BPPV) at the same age group and from the same period were selected as the control group.To study the possible risk factors of cerebral infarction using logistic regression analysis.Results The increasing HCY enlarged the CI risk before inclusion of FA and VitB12 into the analysis.After inclusion of those factors,the correlation between increasing HCY and the enlarged CI risk disappeared.Instead,the decreasing FA and VitB12 increased the CI risk.Conclusion The relationship between HCY and CI was affected by FA and VitB12.
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Adhesion G protein-coupled receptors(aGPCRs) play a significant role in cognitive impairment related diseases.As an important member of aGPCRs,brain-specific angiogenesis inhibitor 1 (BAI1) has a prominent impact on anti-angiogenesis,anti-tumor and participating in immune phagocytosis.Recent research found out that BAI1 exerts a great influence on synaptogenesis and synaptic plasticity,but few studying concerning BAI1 in nervous system.Nowadays,the aging of population aggravates the occurrence of cognitive impairment.The pathogenesis of Alzheimer's disease and vascular cognitive impairment remains elusive,and identification of cognitive impairment at an early stage faces challenges.In the stage of mild cognitive impairment,synaptic damage is evident.BAI1 can regulate the function of postsynaptic membrane,synaptogenesis,synaptic signal transmission and the morphological development of dendritic spines.Therefore,it may potentially act as an early-warning index and intervention target for cognitive impairment.
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Adhesion G protein-coupled receptors(aGPCRs) play a significant role in cognitive impairment related diseases. As an important member of aGPCRs, brain-specific angiogenesis inhibitor 1(BAI1) has a prominent impact on anti-angiogenesis, anti-tumor and participating in immune phagocytosis. Recent research found out that BAI1 exerts a great influence on synaptogenesis and synaptic plasticity, but few studying concerning BAI1 in nervous system. Nowadays, the aging of population aggravates the occurrence of cognitive impairment. The pathogenesis of Alzheimer's disease and vascular cognitive impairment remains elusive, and identification of cognitive impairment at an early stage faces challenges. In the stage of mild cognitive impairment, synaptic damage is evident. BAI1 can regulate the function of postsynaptic membrane, synaptogenesis, synaptic signal transmission and the morphological development of dendritic spines. Therefore, it may potentially act as an early-warning index and intervention target for cognitive impairment.
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Objective To investigate the effect of capsaicin on cognitive function and the expression of TRPV1 protein in hippocampus of rats with chronic cerebral hypoperfusion (CCH).Methods 60 SD rats were randomly divided into control group (SHAM group),chronic cerebral hypoperfusion group (CCH group),placebo control group(PC group) and capsaicin group(CAP group) with 15 in each group.The chronic cerebral hypoperfusion rat model was established by permanent bilateral common carotid artery occlusion.The rats in CAP group and PC group were given capsaicin and saline respectively by intraperitoneal injection,twice a week.The spatial learning and memory ability and emotion of rats were observed by Morris water maze test and open field test,and the expression of TRPV1 in the hippocampus of rats was detected by Western blot.Results (1) In the open field experiment,compared with the SHAM group (22.60±4.60),the standing times of the CCH group(12.10±2.80) decreased (P<0.01),but the standing times of CAP group (19.30± 4.16) increased compared with that of h PC group(12.50 ±2.68) (P<0.01).(2) In Morris water maze test,positioning navigation experiment showed that compared with the SHAM group,the escape latency of the CCH group and the PC group increased (P<0.05),while the escape latency of CAP group was shorter than that of the PC group (P< 0.05).And in the space exploration experiment,compared with the SHAM group (1.87 ± 0.64),the times of crossing the platform in CCH group (0.75 ± 0.89) and the PC group (1.00± 0.93) decreased,while the latency of crossing the platform increased (P<0.01).And the times of crossing the platform in CAP group((2.38±0.74) increased compared with that of PC group,and the latency of crossing the platform of CAP group decreased compared with that of PC group (P<0.01).(3) Results of Western blot showed that compared with the SHAM group,the level of TRPV1 in rat hippocampus of CCH group was down regulated (P<0.05),and the expression of TRPV1 in CAP group was higher than that of PC group (P<0.05).Conclusion Capsaicin can effectively improve cognitive impairment in rats with chronic cerebral hypoperfusion,which may be related to the up-regulation of TRPV1 protein expression in hippocampal tissues.
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Objective To compare the clinical outcomes in elderly stroke patients with large artery atheroclerosis (LAA) and those with cardiogenic embolism (CE)-induced large vessel occlusion (LVO).Methods One hundred and twenty-two ≥65 years old sroke patients with LVO who underwent CT angiography or MR angiography in our hospital from June 2013 to June 2017 were divided into LAA group (n=62) and CE-induced LVO group (n =60) according to the TOAST Classification.Their NIHSS scores on admission and after discharge,good outcome after 3 months of stroke,recurrence rate and mortality of stroke were recorded and compared.Results No significant difference was found in the recurrence rate and mortality of stroke between the two groups (P>0.05).The good outcome rate was significantly higher while the NIHSS score at discharge was significantly lower in CE-induced LVO group than in LAA group (33.33% vs 17.74%,P=0.048;10.50±1.24 vs 11.83±1.53,P=0.001).No significant difference was found in the occlusion sites between the two groups (P>0.05).Conclusion The recovery of neurological function is poorer in stroke patients with LVO than in those with CE-induced LVO.However,no significant difference can be found in the recurrence rate and mortality of stroke between the two groups.
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Objective To observe the effect of monosialoteterahexosyl ganglioside (GM1) combined with Shuxuening injection on nerve function in patients with acute cerebral infarction (ACI) and its mechanism.Methods A total of 94 patients with ACI admitted to the Department of Neurology in Xiangyang First Peoples' Hospital Affiliated to Hubei Medical College from January 2013 to June 2016 were enrolled,and they were divided into two groups by random number table,each group 47 cases.The patients in two groups were all given conventional western medicine treatment;The patients in one group (single group) were treated by intravenous (Ⅳ) drip of GM1,100 mg once a day;and the patients in another group (combined group),by above GM1 Ⅳ drip combined with Shuxuening intramuscular injection,once 2 mL,twice a day;the therapeutic course in two groups was 14 days.Before and after treatment,the changes of China stroke clinical neurological impairment score (CSS score),glasgow coma score (GCS),nerve factor,oxidative stress index and hemodynamics index of two groups were observed.Results Compared with those before treatment,after treatment the CSS score,the levels of neuron specific enolase (NSE),and malondialdehyde (MDA) were significantly lower,while the GCS score,the levels of nerve growth factor (NGF),neurotrophic factor (NTF),maximum blood flow velocity (Vmax),minimum blood flow velocity (Vmin),mean blood flow velocity (Vmean),mean blood flow quantity (Qmean),glutathione peroxidase (GSH-Px),catalase (CAT) and superoxide dismutase (SOD) were all significantly higher in the combined group (all P < 0.05).After treatment,the CSS score,levels of NSE and MDA in the combined group were significantly lower than those of the single group [CSS:11.20 ± 1.78 vs.16.24 ± 1.95,NSE (μg/L):13.17± 1.00 vs.17.68 ± 1.84,MDA (μmol/L):4.14±0.49 vs.5.61 ±0.50,all P < 0.05],GCS score,NGF,NTF,GSH-Px,CAT,SOD,Vmax,Vmin,Vmean and Qmean of the combined group were all significantly higher than those of the single group [GCS:13.68± 1.85 vs.12.01±1.76,NGF (ng/L):88.10±8.83 vs.68.13±7.16,NTF (pg/L):5.13±0.38 vs.3.71±0.30,GSH-Px (U/L):128.13±8.07 vs.103.90±6.58,CAT (U/L):25.74±2.15 vs.19.43± 1.84,SOD (μU/L):94.36±8.00 vs.77.29±7.34,Vmax (cm/min):48.23±3.36 vs.43.17±2.88,Vmin (cm/'min):8.11±0.76 vs.6.85 ± 0.64,Vmean (cm/min):18.69 ± 1.37 vs.15.60 ± 1.24,Qmean (mL/min):9.10 ± 0.74 vs.7.79 ± 0.66,all P < 0.05].Conclusions GM1 combined with Shuxuening injection can improve nerve function in patients with ACI by improving brain blood flow,secreting neurotrophic related factors and inhibiting oxidative stress reaction,thus it has important clinical significance for repairing the damaged nerve function.
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Objective To compare and analyze clinical effects of Bipolar transurethral plasma kinetic enucleation of prostate (PKEP) and transurethral resection of the prostate(TURP) on the treatment huge benign prostatic hyperplasia.Methods Nine-six cases of huge benign prostatic hyperplasia were selected in this hospital from March 2012 to March 2015.All the patients were divided into two groups according to different operation method,namely PKEP group and TURP group.Then the operative time,bleeding amount,bladder washing time,hospital stay,complications between two groups were compared,and the international prostate symptom score (IPSS),quality of life score (QOL),maximal urinary flow rate (Qmax),residual urine volume 6 months before and after operation were compared between the two groups.Results The operative time [(100.0 ± 3.5)min],bleeding amount [(161.0 ± 9.2) mL],bladder washing time[(15.2 ± 1.2) h],hospital stay[(10.8 ± 2.6) d],complications (6 cases) in PKEP group were less than that in the TURP group,which were(132.0±4.2)min,(198.0±12.1)mL,(36.8±1.3)h,(13.6±2.9)d,complications (18 cases)respectively(P<0.05).The IPSS,QOL,Qmax,residual urine volume in both group were significantly improved compared with surgery before(P<0.05),and there were no significant differences between the two groups(P>0.05).Conclusion PKEP and TURP both are effective surgeries for the treatment of huge BPH,while PKEP has short operation time,less intraoperarive bleeding and low incidence of complications,it is worthy of further clinical promotion.
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<p><b>OBJECTIVE</b>To evaluate the value of collateral score based on CT perfusion (CTP-CS) in predicting the clinical outcome of patients with anterior circulation ischemic stroke after thrombectomy.</p><p><b>METHODS</b>Clinical data of acute ischemic stroke patients with anterior artery occlusion undergoing endovascular treatment in the Second Affiliated Hospital, Zhejiang University School of Medicine during October 2013 and October 2016 were retrospectively reviewed. Collateral scores were assessed based on CTP and digital subtraction angiography (DSA) images, respectively. And DSA-CS or CTP-CS 3-4 was defined as good collateral vessels. Good clinical outcome was defined as a modified Rankin Scale (mRS) ≤ 2 at 3 months after stroke. The binary logistic regression model was used to analyze the correlation between the collateral score and clinical outcome, and the receiver operating characteristic (ROC) curve was used to analyze the value of DSA-CS and CTP-CS in predicting the clinical outcome.</p><p><b>RESULTS</b>Among 40 patients, 33 (82.5%) acquired recanalization and 16 (40.0%) got good outcome. Compared with poor outcome group, the collateral score (all<0.05) and the rate of good collateral vessels were higher in good outcome group (all<0.01). After adjust baseline National Institute of Health Stroke Scale (NIHSS) and onset to recanalization time (ORT), good collateral vessels were independent factor of good outcome (CTP-CS:=48.404, 95%:1.373-1706.585,<0.05; DSA-CS:=34.651, 95%:1.147-1047.018,<0.05). Collateral scores based on CTP and DSA had good consistency (=0.697,<0.01), and ROC curve showed that the predictive value of CTP-CS and DSA-CS were comparable (both AUC=0.726, 95%:0.559-0.893,<0.05).</p><p><b>CONCLUSIONS</b>CTP-CS can predict the clinical outcome of patients with anterior circulation ischemic stroke after thrombectomy.</p>
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Objective To research the clinical effect of transurethral plasmakinetic enucleation of prostate (PKEP) in the treatment of high-risk huge benign prostate hyperplasia(BPH).Methods Fifty-two cases of high-risk huge(>120 g) BPH in this hospita1 from May 2010 to May 2015 were selected and performed PKEP.International prostate symptoms score(IPSS),quality of life(QOL) score,residual urine(RUV) and biggest urine flow rate(Qmax) were observed after operation.Results The mean operation time was (130.12 ± 12.14) min,the mean intraroperation bleeding amount was (120.24±9.81) mL,the mean hospital stay was (14.52 ± 1.82)d,the mean weight of resected prostate tissues was (113.42 ± 12.53)g.Follow-up lasted for 6 months without serious complications.IPSS、QOL,RUV and Qmax after operation were improved obviously,the difference was statistically significant compared with before operation(P<0.05).Conclusion PKEP is safe and effective in the treatment of high-risk huge BPH.
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Objective To construct the Hut78 cell line with EZH2 gene knocked into by CRISPR/Cas9 system. Methods The EZH2 expression vector pMD-18T-EZH2 with homologous arm and the sgRNA expression vector pSpCas9 (BB)-2A-Puro-sgRNA, which could cut the double stranded genomic DNA, were constructed, and the two vectors were co-transfected into Hut78 cells. Then the expression of EZH2 mRNA was detected by qPCR, and the expressions of EZH2 and H3K27me3 proteins were detected by Western blot assay. Results The pMD-18T-EZH2 and pSpCas9(BB)-2A-Puro-sgRNA recombinant vectors were confirmed by DNA sequencing. When Hut78 cells were transfected with the two recombinant plasmid, qPCR results showed that the expression of EZH2 mRNA was significantly increased, and Western blot analysis showed that the expressions of EZH2 and H3K27me3 proteins were significantly increased. Conclusion EZH2 gene is successfully knocked into Hut78 cells by CRISPR/Cas9 system.
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Objective To investigate the 4-year follow up of cognitive function outcomes and characteristics in patients after stroke.Methods Sixty three cases according with the diagnostic standard of acute unifocal subcortical stroke were consecutively collected in neurological ward from December 2009 to November 2010.They were followed up for average four years.Forty one out of them completed the neuropsychology tests identical to the baseline,which covered the general cognition function,attention,execution,memory,language,spatial,etc.According to the standard of clinical diagnosis,cognition function is divided into five degrees,including normal,VCI-ND,mild VaD,moderate VaD,and severe VaD.The improved group had 13 cases whose cognition function was improved by one or more ranks.The progressive group had 12 cases whose cognition function progressed by one or more ranks.The stable group had 16 cases whose cognition function remained the same as the baseline.Results According to qualitative analysis on the baseline versus 4-year follow-up outcome,in 13 improved cases,8 were VCI-ND and 5 were mild VaD.In 16 stable cases,11 were normal,4 were VCI-ND and 1 was mild VaD.In 12 progressive cases,3 were normal (change to mild VaD after follow-up),5 were VCI-ND (change to mild and moderate VaD after follow-up) and 4 were mild VaD (change to moderate VaD after 4-year follow-up).In the comparison of baseline cognition function among the improved,progressive and stable group,there was only one significantly different score (the right number of SCWT-A) in the improved and progressive group.The cognition function of improved group had significant differences in CFT-copy,right number of SCWT-C and the time of TMT-B before versus after follow-up.The cognition function of progressive group had significant differences in AVLT-Delay Recall and CFT-Recall.Conclusion Long-term cognitive function outcome after stroke is heterogenetic.The location of cognitive impairment or progression is not the same model for different cognitive outcome group.
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Microglia play a pivotal role in clearance of Aβ by degrading them in lysosomes, countering amyloid plaque pathogenesis in Alzheimer's disease (AD). Recent evidence suggests that lysosomal dysfunction leads to insufficient elimination of toxic protein aggregates. We tested whether enhancing lysosomal function with transcription factor EB (TFEB), an essential regulator modulating lysosomal pathways, would promote Aβ clearance in microglia. Here we show that microglial expression of TFEB facilitates fibrillar Aβ (fAβ) degradation and reduces deposited amyloid plaques, which are further enhanced by deacetylation of TFEB. Using mass spectrometry analysis, we firstly confirmed acetylation as a previously unreported modification of TFEB and found that SIRT1 directly interacted with and deacetylated TFEB at lysine residue 116. Subsequently, SIRT1 overexpression enhanced lysosomal function and fAβ degradation by upregulating transcriptional levels of TFEB downstream targets, which could be inhibited when TFEB was knocked down. Furthermore, overexpression of deacetylated TFEB at K116R mutant in microglia accelerated intracellular fAβ degradation by stimulating lysosomal biogenesis and greatly reduced the deposited amyloid plaques in the brain slices of APP/PS1 transgenic mice. Our findings reveal that deacetylation of TFEB could regulate lysosomal biogenesis and fAβ degradation, making microglial activation of TFEB a possible strategy for attenuating amyloid plaque deposition in AD.
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Animais , Humanos , Camundongos , Doença de Alzheimer , Metabolismo , Patologia , Peptídeos beta-Amiloides , Metabolismo , Precursor de Proteína beta-Amiloide , Genética , Metabolismo , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos , Química , Genética , Metabolismo , Encéfalo , Metabolismo , Células Cultivadas , Canais de Cloreto , Genética , Metabolismo , Modelos Animais de Doenças , Células HEK293 , Lisossomos , Genética , Metabolismo , Camundongos Transgênicos , Microglia , Biologia Celular , Metabolismo , Mutagênese Sítio-Dirigida , Peptídeos , Química , Ligação Proteica , Interferência de RNA , Sirtuína 1 , Genética , MetabolismoRESUMO
Objective To establishing an immortal cell line of familial papillary thyroid carcinoma (FPTC), and explore a new approach for studying familial non-medullary thyroid carcinoma (FNMTC). Methods The specimen from a patient with FPTC was selected, separated, and the primary cells were cultured using DMEM/F12 medium (with TSH, T3, EGF and hydrocortisone). To inducing cell immortalization, the exogenous genes SV40T/TERT were transfected into cells by two ways. RT-PCR was used to detect the expressions of thyroid peroxidase (TPO), thyroid globulin (TG), thyroid stimulating hormone receptor (TSHR) and sodium/iodide co-transporter (NIS). Immunofluorescence method was used to detect the expressions of TPO and GPC3. In order to detect the genomic mutations, the peripheral blood DNA of the patient was extracted. The cell genome was detected. Results The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells can stably grow for six months, FPTC-S (with SV40T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST (with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST can stably express TPO, TG and TSHR in mRNA level. MLH1 R217C mutation existed in the peripheral blood of the patient, and BRAF V600E mutation existed in the primary cultured cells. Either the primary or the immortal cells showed MLH1 R217C mutation. Conclusion This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH 1 R217C and BRAF V600E mutations. This cell line provides a research model for studying these mutations in FPTC.
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Objective To explore the influence of high fat diet on learning and memory,as well as the alteration of the number of neurons and morphology of dendritic spines in rat hippocampi.Methods 24 male adult SD rats were randomly assigned to high fat diet group or control group.The rats were fed with high-fat diet or standaM laboratory rodent chow diet for 12 weeks.Learning and memory were tested by Morris water maze and object recognition tests, and mood and motor ability were tested by open field tests.Golgi staining detected dendritic spine density of hippocampal neurons, and Nissl staining was used to observe the number of hippocampal neurons and pathological changes.Results High-fat diet induced rat spatial learning deficits, which was demonstrated by the prolonged escape latency ((38.50±9.70) s, (20.08±7.35) s, (19.96± 10.56) s, (22.75± 12.51) s, (14.56±4.82) s) compared with the control ((33.61±12.41) s, (14.25±7.89) s, (15.06±7.59) s, (5.53±2.81) s, (4.7± 1.58) s).The spatial memory deficits demonstrated that the latency reaching platform ((30.46± 21.43) s) was prolonged compared with control ((5.18± 1.33)s).The working memory was impaired, which was demonstrated by the prolonged escape latency compared with control group (P< 0.05).Discrimination index lowered than control group ((0.67±0.12) vs (0.81±0.08)), and the difference was significant (P=0.038), but no anxiety behaviors were observed(P=0.461).The neuron number of hippocampal neurons and dendritic spine density were significantly lowered than those in the control group((209.73±24.29) vs (262.2±18.94), (17.9±2.84) vs (21.93±2.56) ,respectively) (P<0.05).Conclusion Intake of high-fat diet can impair learning and memory in rats, as well as decrease the number of neurons and the density of dendritic spines in the hippocampus.
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Objective The aim of this study was to investigate the prevalence of erectile dysfunction (ED) and to assess the risk factors of ED in male post‐stroke patients .Methods One hundred and twenty six male post‐stroke patients were invited to par‐ticipate in this study .Patients were divided into ED group and non‐ED group according to the erectile function evaluated by Interna‐tional Index of Erectile Function‐5 (IIEF‐5) .The neurological impairment was measured using the National Institute of Health Stroke Scale (NIHSS) and the modified Rankin Scale (mRS) .Depressive symptoms was measured using the Hamilton Depression Scales (HAMD) .The global cognitive function was measured using the Mini mental Status Examination (MMSE) .The clinical and psychosocial factors were compared between the ED and non‐ED groups .Univariate and multivariate Logistic regression analyses were employed to assess the risk factors with ED .Results There were 38 patients (30 .2% ) with ED in all the 126 male post‐stroke patients .Univariate Logistic regression analyses revealed that hypertension ,diabetes ,ACEI ,and depression were significantly related to ED in male post‐stroke patients (P<0 .05) .Multivariate Logistic regression analyses revealed that ACEI and depression were independently associated with ED in male post‐stroke patients (P<0 .05) .Conclusion ED is common in Chinese male post‐stroke patients .ACEI and depression are the major determinants of ED .
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<p><b>OBJECTIVE</b>To assess the relationship between protease-activated receptor 1 (PAR1) expression in the basilar artery and cerebral vasospasm (CVS) in a rat model of subarachnoid hemorrhage (SAH).</p><p><b>METHODS</b>Twenty-four SD rats were randomized into normal control group, SAH 3-days group, SAH 5-days group and SAH 7-days group. Rat models of SAH were established by two injections of blood into the cisterna magna and the behavioral changes of the rats were observed. The basilar arteries were taken at 3, 5, or 7 days following the modeling for measuring the cross-sectional area of the basilar artery and for immunohistochemical detection of PAR1 expression.</p><p><b>RESULTS</b>The SAH model rats, especially those in SAH 3-days group, presented with obvious neurological deficits, which was not found in the normal control group. CVS was not observed in the normal control group but occurred in the SAH model rats, which showed reduced cross-sectional area of the basilar artery and worsening spasm over time. The expression level of PAR1 tended to increase gradually in SAH 3-days, SAH 5-days and SAH 7-days groups. Pearson correlation analysis showed an inverse correlation between the expression of PAR1 and the cross-sectional area of the basilar artery (r=-0.779, P<0.01).</p><p><b>CONCLUSIONS</b>The expression of PAR1 increases significantly in rat basilar artery wall following SAH in positive correlation with the severity of CVS, suggesting the role of thrombin in the pathological process of CVS after SAH.</p>
Assuntos
Animais , Ratos , Artéria Basilar , Metabolismo , Ratos Sprague-Dawley , Receptor PAR-1 , Metabolismo , Hemorragia Subaracnóidea , Metabolismo , Vasoespasmo Intracraniano , MetabolismoRESUMO
<p><b>BACKGROUND</b>Based on the excellent medical care and management system for Chinese veterans, as well as the detailed medical documentation available, we aim to construct a Chinese Veteran Clinical Research (CVCR) platform on non-communicable diseases (NCDs) and carry out studies of the primary disabling NCDs.</p><p><b>METHODS</b>The Geriatric Neurology Department of Chinese People's Liberation Army General Hospital and veterans' hospitals serve as the leading and participating units in the platform construction. The fundamental constituents of the platform are veteran communities. Stratified typical cluster sampling is adopted to recruit veteran communities. A cross-sectional study of mental, neurological, and substance use (MNS) disorders are performed in two stages using screening scale such as the Mini-Mental State Examination and Montreal cognitive assessment, followed by systematic neuropsychological assessments to make clinical diagnoses, evaluated disease awareness and care situation.</p><p><b>RESULTS</b>A total of 9 676 among 277 veteran communities from 18 cities are recruited into this platform, yielding a response rate of 83.86%. 8 812 subjects complete the MNS subproject screening and total response rate is 91.70%. The average participant age is (82.01±4.61) years, 69.47% of veterans are 80 years or older. Most participants are male (94.01%), 83.36% of subjects have at least a junior high school degree. The overall health status of veterans is good and stable. The most common NCD are cardiovascular disorders (86.44%), urinary and genital diseases (73.14%), eye and ear problems (66.25%), endocrine (56.56%) and neuro-psychiatric disturbances (50.78%).</p><p><b>CONCLUSION</b>We first construct a veterans' comprehensive clinical research platform for the study of NCDs that is primarily composed of highly educated Chinese males of advanced age and utilize this platform to complete a cross-sectional national investigation of MNS disorders among veterans. The good and stable health condition of the veterans could facilitate the long-term follow-up studies of NCDs and provide prospective data to the prevention and management of NCDs.</p>