Automatic radiolabeling of the norepinephrine transporter targeted tracer 18F-mFBG and evaluation of 18F-mFBG PET/CT imaging in pheochromocytoma / 中华核医学与分子影像杂志

Objective:To fulfill the automatic radiolabeling of the norepinephrine transporter (NET) trancer 18F-meta-fluorobenzylguanidine (mFBG), and explore the 18F-mFBG PET/CT imaging effect of pheochromocytoma. Methods:On the basis of the chemical structure of mFBG, a spirocyclic iodonium ylide was used as the precursor to undergo a 3-step reaction sequence (radiofluorination, deprotection and neutralization) on AllinOne synthesis module. Purification by high performance liquid chromatography and formulation were conducted to generate 18F-mFBG. The corresponding quality control tests of 18F-mFBG product was performed. Afterwards, a postoperative patient with pheochromocytoma underwent 18F-mFBG PET/CT imaging. Results:The radiosynthesis was accomplished within 70 min, and 18F-mFBG was obtained in (17.8±2.4)% non-decay-corrected radiochemical yield ( n=5), with radiochemical purity >97% and molar activity >59.2 GBq/μmol. Sterility test, bacterial endotoxins test, abnormal toxicity test and the acetonitrile residue all met the requirements of Pharmacopoeia of the People′ s Republic of China (2020 Volume Ⅳ). The 18F-mFBG PET/CT imaging disclosed high uptake in pheochromocytoma and clear localization of lesions. Conclusions:The automatic radiolabeling of the NET targeted tracer 18F-mFBG is successfully realized by commercially available synthesis module, and the production quality meets all requirements for clinical translation. 18F-mFBG has a potential to image neuroendocrine lesions in clinical setting.

Characterization of the nigroestriatal system in a sample of patients with amyotrophic lateral sclerosis / Caracterização do sistema nigroestriatal em uma amostra de pacientes com esclerose lateral amiotrófica

Abstract Background The coexistence of amyotrophic lateral sclerosis (ALS) with clinical forms of Parkinson disease (PD), although uncommon, is found to a greater degree than one would expect by chance. The pathological mechanisms of ALS and PD are still not fully understood, and the coexistence of these two diseases suggests that they could share mechanisms in common. Objective Here we present a sample of patients with clinically definitive or probable ALS who were evaluated with single-photon emission computed tomography SPECT/TRODAT and compared with non-ALS controls. Methods Patients with clinically definite or probable ALS were assessed with the amyotrophic lateral sclerosis functional rating scale (ALSFRS) to define severity and had their demographic data collected. The TRODAT results of patients with ALS were compared with those of patients with a diagnosis of PD with less than 10 years of duration, and with patients with a diagnosis of others movement disorders not associated with neurodegenerative diseases. Results A total of 75% of patients with ALS had TRODAT results below the levels considered normal; that was also true for 25% of the patients in the control group without neurodegenerative disease, and for 100% of the patients in the PD group. A statistically significant difference was found between patients with ALS and the control group without neurodegenerative disease in the TRODAT values < 0.05. Conclusions Our study fits with the neuropathological and functional evidence that demonstrates the existence of nigrostriatal dysfunction in patients with ALS. Further research to better understand the role of these changes in the pathophysiological process of ALS needs to be performed.

Resumo Antecedentes A coexistência da esclerose lateral amiotrófica (ELA) com formas clínicas da doença de Parkinson (DP), embora incomum, é encontrada em um grau maior do que seria esperado ao acaso. Os mecanismos patológicos da ELA e da DP ainda não são totalmente compreendidos e a coexistência dessas duas doenças sugere que elas podem compartilhar mecanismos em comum. Objetivo Apresentamos uma amostra de pacientes com ELA clinicamente definida ou provável que foram avaliados com tomografia computadorizada por emissão de fóton único (SPECT)/TRODAT e comparados com controles sem ELA. Métodos Pacientes com ELA clinicamente definida ou provável foram avaliados com a escala funcional de esclerose lateral amiotrófica (ALSFRS) para definir a gravidade e foram coletados os seus dados demográficos. Os resultados do TRODAT de pacientes com ELA foram comparados com aqueles de pacientes com diagnóstico de DP com menos de 10 anos de duração e com pacientes com diagnóstico de outros distúrbios do movimento não associados a doenças neurodegenerativas. Resultados Um total de 75% dos pacientes com ELA apresentou resultados de TRODAT abaixo dos níveis considerados normais; 25% no grupo controle sem doença neurodegenerativa e 100% no grupo DP. Uma diferença estatisticamente significativa foi encontrada entre os pacientes com ELA e o grupo controle sem doença neurodegenerativa nos valores de TRODAT p< 0,05. Conclusões Nosso estudo está de acordo com as evidências neuropatológicas e funcionais que demonstram a existência de disfunção nigroestriatal em pacientes com ELA. Mais pesquisas para entender melhor o papel dessas mudanças no processo fisiopatológico da ELA precisam ser realizadas.

Beta-adrenergic pathway activation in head and neck cancer: a comprehensive analysis of the clinical and genomic features using integrated bioinformatics / Ativação da via beta-adrenérgica no câncer de cabeça e pescoço: uma análise abrangente das características clínicas e gênomicas utilizando a bioinformática

O estresse crônico leva à ativação da via de sinalização beta-adrenérgica. Sua ativação tem sido implicada na progressão de diferentes tipos de câncer, mas seu papel nos carcinomas espinocelulares de cabeça e pescoço (CECPs) permanece indefinido. O objetivo deste estudo foi investigar o papel da ativação da via betaadrenérgica na progressão dos CECPs, avaliar seu impacto na sobrevida dos pacientes e buscar possíveis terapias para pacientes que encontravam-se com a via beta-adrenérgica ativa. Quinhentos e vinte pacientes do The Cancer Genome Atlas com CECPs primários foram divididos em dois grupos: ADRB2baixa / SLC6A2baixa e ADRB2alta / SLC6A2alta. A associação de características clinicopatológicas e genômicas entre os grupos foram analisadas utilizando bioinformática. Os genes diferencialmente expressos (DEGs) foram identificados através da análise da expressão diferencial. A análise de sobrevida também foi realizada com base nas expressões ADRB2 e SLC6A2. Foram identificados medicamentos em potencial para tratamento de CECPs com base nos DEGs. Houve associação entre as expressões ADRB2 e SLC6A2 com idade, raça, localização do tumor, grau histológico, invasão perineural e status do HPV p16. Foram identificados 898 DEGs entre os grupos. Foi demonstrado que a expressão ADRB2alta / SLC6A2alta influenciou a proliferação, adesão e invasão de células CECPs além da angiogênese. Pacientes com carcinomas espinocelular de laringe e faringe apresentando expressão ADRB2alta / SLC6A2alta tiveram menor sobrevida. Por fim, 56 drogas antineoplásicas e imunoterápicas aprovadas pelo Food Drugs Administration foram identificadas como potenciais alvos para o tratamento personalizado. Significância: Estes achados sugerem fortemente um papel proeminente da sinalização beta-adrenérgica no CECPs ao estimular um fenótipo tumoral mais agressivo. Estas alterações tiveram um impacto negativo no prognóstico dos pacientes com CECP em região de faringe e laringe(AU)

Chronic stress leads to the activation of the beta-adrenergic pathway. Its activation has been implicated in the progression of different types of cancer but its role on head and neck squamous cell carcinomas (HNSCCs) remains undefined. The aim of this study was to investigate the influence of the beta-adrenergic pathway activation in the progression of HNSCCs, assess its impact in the survival of the patients, and explore the potential targets. Five hundred and twenty The Cancer Genome Altas patients with primary HNSCCs were divided in two groups: ADRB2low / SLC6A2low and ADRB2high / SLC6A2high. The association of clinicopathological and genomic features between the groups was analyzed using a bioinformatic approach. Differentially expressed genes (DEGs) were identified through differential expression analysis. Survival analysis was also performed based on ADRB2 and SLC6A2 expressions. Potential drugs for treatment of HNSCC were identified based on the DEGs. There was association between ADRB2 and SLC6A2 expressions with age, race, tumor site, histologic grade, perineural invasion, and HPV p16 status. It was identified 898 DEGs between the groups. It was demonstrated that ADRB2high / SLC6A2high expression influenced HNSCC cells proliferation, adhesion, invasion, and angiogenesis. Patients with larynx and pharynx squamous cell carcinomas presenting ADRB2high / SLC6A2high expression showed had lower survival rates. Finally, 56 Food Drugs Administration-approved antineoplastic and immunotherapeutic drugs were identified as potential targets for the personalized treatment. Significance: These findings strongly suggest a prominent role of beta-adrenergic pathway in HNSCC by stimulating a more aggressive tumoral phenotype. These alterations were shown to negatively impact the prognosis of patients with larynx and pharynx squamous cell carcinomas(AU)

Correlations between striatal dopamine transporter distribution, glucose metabolism and clinical symptoms in Parkinson′s disease / 中华核医学与分子影像杂志

Objective@#To investigate the correlations among striatal dopamine transporter (DAT) distribution, glucose metabolism and Parkinson′s disease (PD) clinical symptoms.@*Methods@#Twenty-five clinically confirmed idiopathic PD patients (17 males, 8 females, age: (59.8±9.2) years) who underwent 11C-2-beta-carbomethoxy-3-beta-(4-fluorophenyl)tropane (CFT) and 18F-fluorodeoxyglucose (FDG) PET imaging from January 2015 to December 2016 were reviewed. The detailed clinical scores were systematically collected from all patients. Correlations between DAT distribution, glucose metabolism and clinical symptoms were evaluated at global and voxel levels using Pearson correlation analysis.@*Results@#There were significantly positive correlations between the PD-related pattern (PDRP) value and unified PD rating scale (UPDRS) motor scores, non-motor symptoms scale (NMSS) scores, activity of daily living scale (ADL) scores (r values: 0.580, 0.522, 0.557, all P<0.05). The CFT uptake of ipsilateral caudate nucleus, anterior putamen, and posterior putamen were negatively correlated with UPDRS motor scores (r values: -0.496, -0.492, -0.457, all P<0.05), while those had no significant correlations with NMSS scores (r values: -0.420, -0.402, -0.355, all P>0.05). The CFT uptake of ipsilateral caudate nucleus and anterior putamen were negatively correlated with ADL scores (r values: -0.502, -0.522, both P<0.05). There were no significant correlations between CFT uptake in contralateral striatal, anterior putamen, posterior putamen and PDRP values, UPDRS motor scores, NMSS scores and ADL scores(r values: from -0.466 to -0.129, all P>0.05). The presence of the significant correlations between UPDRS motor scores, ADL scores and the CFT radioactive count were confirmed in left caudate nucleus and left putamen (r values: from -0.90 to -0.47, all P<0.05). The metabolic PET imaging disclosed a set of brain regions correlating with the clinical symptoms. The presence of significant correlations between the metabolic PET imaging and CFT uptake were confirmed in bilateral caudate nucleus (r values: 0.47-0.90, both P<0.01), precentral gyrus and insula (r values: -0.90 to -0.47, all P<0.01).@*Conclusion@#The correlations between DAT distribution, glucose metabolism and PD clinical symptoms are complicated, which promote the understanding in the proper application of dopaminergic and metabolic PET imaging in PD and offer more evidences in PD pathophysiological mechanisms.

Formation mechanism of tumor acidic microenvironment and its effects on tumor progression / 肿瘤

Tumor microenvironment plays an important role in the formation and development of tumors, and the acidity is one of the remarkable features of tumor microenvironment. Due to excessive proliferation of tumor cells and abnormal structure of tumor blood vessels, the tumor tissues are usually in a hypoxia state, leading to changes in the metabolic processes of tumor cells. Compared with the normal tissues which mainly rely on aerobic oxidation to obtain energy, the metabolism of tumor cells dominatingly depends on the anaerobic glycolysis. Therefore, the lactate acid produced by glycolysis and the carbon dioxide produced by respiration together result in the acidification of tumor microenvironment (TME), and affects many aspects of tumorigenesis and development. This review focuses on the formation mechanism of tumor acidic microenvironment and its impacts on tumor progression, including tumor immunity, invasion, autophagy and resistance to anti-cancer treatment.

Carnitine-acylcarnitine translocase deficiency with homozygous mutation of c.199-10T＞G: a case report and literature review / 中华围产医学杂志

Objective To investigate the clinical features of carnitine-acylcarnitine translocase deficiency (CACTD) with c.199-10T＞G homozygous mutation and the characteristics of SLC25A20 gene mutation.Methods This study retrospectively analyzed the clinical data,biochemical and genetic features,treatment and outcome of a boy with CACTD with c.199-10T＞G homozygous mutation,who admitted to Guangzhou Women and Children's Medical Center in September 2017.Pertinent articles were retrieved from China National Knowledge Infrastructure (CNKI),Wanfang Database,National Center for Biotechnology Information and PubMed from the establishment of these databases to April 2018 using key words including CACT,SLC25A20 and carnitine-acylcarnitine translocase.Clinical information of all affected cases in the retrieved publications was analyzed.Results (1) The full-term boy born vaginally at a local hospital was transferred to neonatal intensive care unit (NICU) of Guangzhou Women and Children's Medical Center at 2 days and 5 hours due to groaning,cyanosis and severe hypoglycemia (0.8 mmol/L) at 15 h after birth.His elder brother with similar symptoms died of unknown reason at 50 days of age.In this case,ammonemia,liver enzyme and creatine kinase were significantly elevated,amino acid analysis suggested liver damage,and high amounts of dicarboxylic aciduria,low free carnitine,markedly increased long-chain acylcarnitine and hypoketotic hypoglycemia were also observed.His electrocardiogram showed atrioventricular block and ventricular tachycardia.After a series of treatments,including repeated electrical cardioversion,lidocaine and amiodarone for arrhythmia,arginine for blood ammonia level reduction,formula supplement containing L-carnitine and medium-chain fatty acid,the patient whose conditions had significantly improved and was discharged at the request of his parents at 29 days old.Two weeks later,he was re-admitted due to diarrhea,and discharged two days later when he was 45 days old.He was lost to follow up since then.(2) A homozygous mutation of c.199-10T＞G was detected in this boy in SLC25A20 gene,which was also carried by his parents.(3) Thirtytwo publications in English were retrieved,involving 50 cases of CACTD and 100 sequenced alleles.A total of 40 mutations in SLC25A20 gene were found so far,and c.199-10T＞G was the most common mutation with a frequency of 22/100.It was identified in 13 patients,including nine homozygous mutations and four compound heterozygous mutations.Symptoms presented within 72 h after birth (25 min-52 h) in all the 13 infants,such as hypoketotic hypoglycemia,hyperammonemia,elevated liver enzyme and creatine kinase,significantly decreased free carnitine level,markedly increased level of long-chain acylcarnitine,dicarboxylic aciduria,arrhythmia and cardiomyopathy.The mortality rate of CACTD was 11/12 (the outcome of one case was not reported).Conclusions c.199-10T＞G is the most common SLC25A20 gene mutation reported in Asia population with severe phenotypes and poor outcomes.Early diagnosis and timely treatment of CACTD are crucial.Inborn metabolic diseases such as CACTD should be considered if unexplainable exacerbation of clinical signs in neonatal period,or sudden infant death occurs.

Preparation and imaging of 18 F-VUIIS1008 targeting TSPO in rheumatoid arthritis model / 中华核医学与分子影像杂志

Objective To synthesize a novel 18 F labeled probe targeting translocator protein ( TSPO) ligand 2-( 5, 7-diethyl-2-( 4-( 2-fluoroethoxy ) phenyl ) pyrazolo [ 1, 5-a ] pyrimidin-3-yl )-N, N-diethylacet-amide (VUIIS1008), and evaluate its biodistribution and imaging in rheumatoid arthritis (RA) model. Methods The tosylate substrate was labeled with 18 F using a tosyloxy for fluorine nucleophilic aliphatic substitution to obtain 18 F-VUIIS1008. The labeling efficiency, radiochemical purity, and stability in vitro were determined. In vitro cellular uptake and competitive binding assay were performed on RAW264.7 mac-rophage cells. Biodistribution and microPET/CT imaging were investigated on RA mice established by Com-plete Freund's Adjuvant. Two-sample t test was used to analyze the data. Results 18 F-VUIIS1008 was syn-thesized with the labeling yield up to (41.00±5.00)％, the radiochemical purity>98.00％, and the specific radioactivity >1. 52 × 108 MBq/mmol. 18 F-VUIIS1008 was highly stable with the radiochemical purity >98. 00％ at 4 h after incubation in mouse serum. In vitro, it also exhibited high specific TSPO binding in RAW264.7 macrophage cells. The uptake ratio was (14.00±0.30)％ at 1 h after incubation, and decreased significantly ((4.00±0.70)％;t=12.894, P<0.05) after adding excessive unlabeled VUIIS1008. The half maximal inhibitory concentration (IC50) of 18F-VUIIS1008 binding to TSPO was 0.05 nmol/L in RAW264.7 macrophage cells. In vivo distribution results showed that the uptake of 18 F-VUIIS1008 in the left arthritic ankles reached the peak value of (1.33±0.02) percentage activity of injection dose per gram of tissue (％ID/g) at 1 h after injection. The radioactivity ratio of left ankle arthritic tissue to blood ( A/B) and to normal muscle ( A/M) was 4.40±0.22 and 1.65±0.07 respectively. MicroPET/CT imaging demonstrated that 18F-VUIIS1008 could specifically target and retained in the inflammation site. Conclusion 18 F-VUIIS1008 can be easily synthe-sized with high radiochemical purity and can clearly visualized in RA imaging with low background, suggesting its potential as a novel promising molecular probe targeting TSPO for RA PET imaging.

Correlations between striatal dopamine transporter distribution, glucose metabolism and clinical symptoms in Parkinson's disease / 中华核医学与分子影像杂志

Objective To investigate the correlations among striatal dopamine transporter (DAT) distribution,glucose metabolism and Parkinson's disease (PD) clinical symptoms.Methods Twenty-five clinically confirmed idiopathic PD patients (17 males,8 females,age:(59.8± 9.2) years) who underwent 11 C-2-beta-carbomethoxy-3-beta-(4-fluorophenyl) tropane (CFT) and 18 F-fluorodeoxyglucose (FDG) PET imaging from January 2015 to December 2016 were reviewed.The detailed clinical scores were systematically collected from all patients.Correlations between DAT distribution,glucose metabolism and clinical symptoms were evaluated at global and voxel levels using Pearson correlation analysis.Results There were significantly positive correlations between the PD-related pattern (PDRP) value and unified PD rating scale (UPDRS) motor scores,non-motor symptoms scale (NMSS) scores,activity of daily living scale (ADL) scores (r values:0.580,0.522,0.557,all P＜0.05).The CFT uptake of ipsilateral caudate nucleus,anterior putamen,and posterior putamen were negatively correlated with UPDRS motor scores (r values:-0.496,-0.492,-0.457,all P＜0.05),while those had no significant correlations with NMSS scores (r values:-0.420,-0.402,-0.355,all P＞0.05).The CFT uptake of ipsilateral caudate nucleus and anterior putamen were negatively correlated with ADL scores (r values:-0.502,-0.522,both P＜0.05).There were no significant correlations between CFT uptake in contralateral striatal,anterior putamen,posterior putamen and PDRP values,UPDRS motor scores,NMSS scores and ADL scores(r values:from-0.466 to-0.129,all P＞0.05).The presence of the significant correlations between UPDRS motor scores,ADL scores and the CFT radioactive count were confirmed in left caudate nucleus and left putamen (r values:from-0.90 to-0.47,all P＜0.05).The metabolic PET imaging disclosed a set of brain regions correlating with the clinical symptoms.The presence of significant correlations between the metabolic PET imaging and CFT uptake were confirmed in bilateral caudate nucleus (r values:0.47-0.90,both P＜0.01),precentral gyrus and insula (r values:-0.90 to-0.47,all P＜0.01).Conclusion The correlations between DAT distribution,glucose metabolism and PD clinical symptoms are complicated,which promote the understanding in the proper application of dopaminergic and metabolic PET imaging in PD and offer more evidences in PD pathophysiological mechanisms.

Effect of carbon monoxide postconditioning on pyroptosis induced by oxygen-glucose deprivation and restoration in rat hippocampal neurons: the relationship with mPTP/ROS signaling pathway / 中华麻醉学杂志

Objective To evaluate the effect of carbon monoxide (CO) postconditioning on pyroptosis induced by oxygen-glucose deprivation and restoration (OGD/R) in rat hippocampai neurons and the relationship with mitochondrial permeability transition pore (mPTP)/reactive oxygen species (ROS) signaling pathway.Methods Primary hippocampal neurons were cultured in vitro,seed in 6-well or 96-well plates,and divided into 5 groups (n =24 each) using a random number table method:control group (C group),OGD/R group,CO postconditioning group (CO group),specific mPTP opener atractyloside plus CO postconditioning group (ACO group),and specific ROS inducer antimycin A plus CO postconditioning group (KCO group).Neurons were subjected to O2-glucose deprivation (OGD) for 16 h followed by restoration of O2-glucose supply for 24 h to establish the model of OGD/R injury.In group CO,neurons were exposed to 2％ CO-5％ CO2 for 3 h at 37 ℃ starting from the end of OGD,followed by normal culture for 21 h.In ACO and KCO groups,atractyloside 20 μmol/L and antimycin A 50 μmol/L were added at the end of OGD,respectively,and the other treatments were similar to those previously described in group CO.Neuronal pyroptosis rate was determined using double immunofluorescent staining cleaved caspase-1-AlexaFluor 568/DAPI after the end of treatments in each group.The neuronal survival rate was determined by MTT,opening of mPTP by Calcein-AM fluorescence,ROS content by DCFH-DA,and expression of interleukin1beta (IL-1β) and IL-18 by Western blot.Results Compared with C group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in the other groups (P＜0.05).Compared with OGD/R group,neuronal pyroptosis rate,ROS content and opening of mPTP were significantly decreased,the neuronal survival rate was increased,and the expression of IL-1β and IL-18 was down-regulated in CO,ACO and KCO groups (P＜0.05).Compared with CO group,neuronal pyroptosis rate and ROS content were significantly increased,the neuronal survival rate was decreased,and the expression of IL-1β and IL-18 was up-regulated in ACO and KCO groups,and opening of mPTP was significantly inctreased in ACO group (P＜0.05).Conclusion CO postconditioning can inhibit OGD/R-induced pyroptosis in rat hippocampal neurons,and the mechanism is related to inhibiting mPTP/ROS signaling pathway.

Effect of rs3910105 in the Synuclein Gene on Dopamine Transporter Availability in Healthy Subjects

PURPOSE: The present study investigated associations between dopamine transporter (DAT) availability and α-synuclein levels in cerebrospinal fluid, as well as synuclein gene (SNCA) transcripts, and the effect of single nucleotide polymorphism of SNCA on DAT availability in healthy subjects. MATERIALS AND METHODS: The study population comprised healthy controls who underwent 123I-FP-CIT single-photon emission computed tomography screening. Five SNCA probes were used to target the boundaries of exon 3 and exon 4 (SNCA-E3E4), transcripts with a long 3′UTR region (SNCA-3UTR-1, SNCA-3UTR-2), transcripts that skip exon 5 (SNCA-E4E6), and the rare short transcript isoforms that comprise exons 1–4 (SNCA-007). RESULTS: In total, 123 healthy subjects (male 75, female 48) were included in this study. DAT availability in the caudate nucleus (p=0.0661) and putamen (p=0.0739) tended to differ according to rs3910105 genotype. In post-hoc analysis, DAT availability in the putamen was lower in subjects of TT genotype than those of CC/CT (p=0.0317). DAT availability in the caudate nucleus also showed a trend similar to that in the putamen (p=0.0597). Subjects of CT genotype with rs3910105 showed negative correlations with DAT availability in the putamen with SNCA-E3E4 (p=0.037, rho=−0.277), and SNCA-E4E6 (p=0.042, rho=−0.270), but not those of CC/TT genotypes. CONCLUSION: This is the first study to investigate the association of rs3910105 in SNCA with DAT availability. rs3910105 had an effect on DAT availability, and the correlation between DAT availability and SNCA transcripts were significant in CT genotypes of rs3910105.

Effects of aldosterone on the protein expressions and the urea transport activity of UT-A1 and UT-A3 / 中华肾脏病杂志

Objective To investigate the effects of protein expressions and the urea transport activity of aldosterone on urea transporter A1 (UT-A1) and urea transporter A3 (UT-A3) in HEK293 cells and Xenopus laevis oocytes.Methods (1) Western Blot was used to investigate the protein expressions of UT-A1 and UT-A3.(2) Cell surface biotinylation was used to investigate the protein expressions of UT-A1 and UT-A3 on the cell surface of Xenopus laevis oocytes.(3) 14C-urea transport experiment was conducted to investigate the transport activity of UT-A1 and UT-A3 in Xenopus laevis oocytes.Results (1) Compared with UT-A1 or UT-A3 high expression groups,the total protein levels of UT-A 1 and UT-A3 were all significantly reduced in aldosterone treatment groups (all P ＜ 0.01).(2) Compared with UT-A1 or UT-A3 high expression groups,the levels of protein expression on cell surface were all significantly reduced in aldosterone groups (all P ＜ 0.01).(3) Compared with UT-A1 or UT-A3 high expression groups,14C-urea transport experiment results showed that aldosterone treatment groups had significantly reduced the urea transporter activity of UT-A1 (1 min:94.32±9.044vs 40.68±4.274,P＜0.01,n=6;3 min:165.0±4.7 vs 80.3±0.6,P＜0.01,n=6),and UT-A3 (1 min:204.6± 3.1 vs 176.7± 9.1,P＜0.05,n=6;3 min:371.4 ± 14.9 vs 318.8 ± 12.0,P＜0.05,n=6).Conclusion Aldosterone can directly down-regulate the protein expressions of UT-A1 and UT-A3 in both total protein and cell surface level,which reduces their urea transport activity.

Role of mitochondriai permeability transition pore in reduction of brain injury by sevoflurane post-conditioning in a rat model of hemorrhagic shock and resuscitation / 中华麻醉学杂志

Objective To evaluate the role of mitochondrial permeability transition pore (mPTP)in reduction of brain injury by sevoflurane postconditioning in a rat model of hemorrhagic shock and resuscitation (HSR).Methods Ninety pathogen-free healthy adult male Sprague-Dawley rats,weighing 300-350 g,were divided into 5 groups (n =18 each) using a random number table:sham operation group (group S),group HSR,sevoflurane postconditioning group (group SP),sevoflurane postconditioning plus atractyloside (ATR,a specific mPTP opener) group (group SP + ATR) and ATR group.Hemorrhagic shock was produced by withdrawing 40％ of the total blood volume from the right carotid artery over an interval of 30 min,and 1 h later the animals were resuscitated by infusion of the shed blood via the left jugular vein over 30 min.SP and SP+ATR groups were exposed to 2.4％ sevoflurane for 30 min starting from the onset of reinfusion.In ATR and SP+ATR groups,ATR 5 mg/kg was intravenously injected at 10 min before reinfusion.Six rats in each group were randomly sacrificed at 24 h after the end of autologous blood reinfusion,and the hippocampus was harvested for determination of the expression of Bcl-2 and Bax in hippocampal tissues (by Western blot) and degree of mPTP opening.At 72 h after the end of autologous blood reinfusion,the rest 6 rats in each group were selected and underwent Morris water maze test,and the cognitive function was evaluated.Results Compared with group S,the escape latency was significantly prolonged,the number of crossing the original platform and locomotor distance in the target quadrant were decreased,the expression of Bcl-2 was down-regulated,the expression of Bax was up-regulated,and the degree of mPTP opening was increased in group HSR (P＜0.05).Compared with group HSR,the escape latency was significantly shortened,the number of crossing the original platform and locomotor distance in the target quadrant were increased,the expression of Bcl-2 was up-regulated,the expression of Bax was down-regulated,and the degree of mPTP opening was decreased in group SP (P＜0.05),and no significant change was found in each parameter in ATR and SP+ATR groups (P＞0.05).Compared with group SP,the escape latency was significantly prolonged,the number of crossing the original platform and locomotor distance in the target quadrant were decreased,the expression of Bcl-2 was down-regulated,the expression of Bax was up-regulated,and the degree of mPTP opening was increased in group SP+ATR (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning ameliorates brain injury may be related to inhibiting mPTP opening in a rat model of HSR.

Relationship between remifentanil-induced hyperalgesia and function of nitrated glutamate transpor-tor-1 and glutamine synthetase in spinal cord of rats with incisional pain / 中华麻醉学杂志

Objective To investigate the relationship remifentanil-induced hyperalgesia and func-tion of nitrated glutamate transportor-1 ( GLT-1) and glutamine synthetase ( GS) in the spinal cord of rats with incisional pain. Methods Thirty-two male Sprague-Dawley rats, weighing 260-280 g, aged 2-3 months, in which caudal catheters were successfully implanted, were divided into 4 groups (n=8 each) u-sing a random number table method: control group ( group C) , incisional pain group ( group I) , remifen-tanil group ( group R) and remifentanil plus incisional pain group ( group RI) . Normal saline was intrave-nously infused for 60 min at 0. 1 ml · kg-1 · min-1 in group C. The model of incisional pain was estab-lished, and normal saline was simultaneously infused for 60 min via the tail vein at 0. 1 ml·kg-1 ·min-1 in group I. Remifentanil was infused for 60 min via the tail vein at 1. 0 μg· kg-1 ·min-1 in group R. The model of incisional pain was established, and remifentanil was infused for 60 min via the tail vein at 1. 0μg· kg-1 ·min-1 in group RI. The mechanical paw withdrawal threshold ( MWT) and thermal paw with-drawal latency ( TWL) were measured at 24 h before infusion of remifentanil or normal saline ( T0 ) and at 2, 6, 24 and 48 h after infusion ( T1-4 ) . The rats were sacrificed after the last measuremnet of pain thresh-old, and the L4-6 segment of the spinal cord was removed for determination of the expression of GLT-1 and GS (by Western blot) and expression of nitrated GLT-1 (nGLT-1) and nitrated GS (nGS) (by Western blot) . Ratios of nGLT-1∕GLT-1 and nGS∕GS were calculated. Results Compared with group C, the MWT was significantly decreased and TWL was shortened at T1-4 , the expression of GLT-1 and GS was down-regu-lated, the expression of nGLT-1 and nGS was up-regulated, and ratios of nGLT-1∕GLT-1 and nGS∕GS were increased in I, R and RI groups ( P<0. 05) . Compared with group I and group R, the MWT was signifi-cantly decreased and TWL was shortened at T1-4 , the expression of GLT-1 and GS was down-regulated, the expression of nGLT-1 and nGS was up-regulated, and ratios of nGLT-1∕GLT-1 and nGS∕GS were increased in group RI ( P<0. 05) . Conclusion The mechanism of remifentanil-induced hyperalgesia may be related to impaired function of GLT-1 and GS in the spinal cord of rats with incisional pain.

Expression changes of glutamate-aspartate transporter and glutamate transporter-1 in glial cells following spinal cord injury in adult rats / 中华创伤杂志

Objective To investigate the changes and their significance of glutamate-aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1) expressions in glial cells following spinal cord injury (SCI) in adult rats.Methods Twenty-five healthy female adult SD rats were randomly divided into control group (5 rats) and experimental group (20 rats).The contusive spinal cord injury models were prepared at T10 segment in the rats in the experimental group according to the modified Allen's method.At days 1,3,7 and 14 following SCI,five rats were sacrificed by cardiac perfusion and the spinal cord segments adjacent to the epicenter of injury were obtained at each time point after the neurological function of hind limbs was assessed using the modified Tarlov scale.Changes of GLAST and GLT-1 expressions were detected semi-quantitatively using immunofluorescence and computer image analysis system (IPP 6.0).Results (1) Single immunofluorescence:Moderate GLAST expression was found in the control group.The GLAST expression was increased slightly at day 1 after SCI,decreased progressively at days 3 and 7 after SCI,and increased slightly at day 14 after SCI.The GLAST expression in experimental group was significantly lower than those in control group at days 3,7 and 14 after SCI (P ＜ 0.05).Moderate GLT-1 expression was detected in the control group.The expression of GLT-1 was increased slightly at day 1 after SCI,decreased to the lowest at day 3 after SCI,and increased slightly at days 7 and 14 after SCI.The GLT-1 expression in experimental group was significantly lower than those in control group at days 3,7 and 14 after SCI (P ＜0.05).(2) Double immunofluorescence:GLAST expression was found on astrocytes in the control group.The GLAST expression in experimental group was decreased at day 1 after SCI,further decreased progressively at days 3 and 7 after SCI,and started to recover at day 14 after SCI.The coexpressions of GLAST and glial fibrillary acidic protein (GFAP) in experimental group were significantly lower than those in the control group at days 3 and 7 after SCI (P ＜ 0.05).The expression of GLAST was found on microglial cells in the control group.The expression of GLAST in experimental group was increased obviously at day 1 after SCI and increased progressively at days 3-14 after SCI.The coexpressions of GLAST and OX-42 in experimental group were significantly than those in the control group at days 3,7 and 14 after SCI (P ＜ 0.05).(3) Double immunofluorescence:GLT-1 expression was found on astrocytes in the control group.The GLT-1 expression was decreased at day 1 after SCI,further decreased progressively at days 3 and 7 after SCI,and started to recover at day 14 after SCI.The coexpressions of GLT-1 and GFAP were significantly lower than those in the control group at days 3 and 7 after SCI (P ＜ 0.05).The GLT-1 expression was found on microglial cells in the control group.The GLT-1 expression was increased obviously at day 1 after SCI and increased progressively at days 3-14 after SCI.The coexpressions of GLT-1 and OX-42 were significantly higher than those in the control group at days 1,3,7 and 14 after SCI (P ＜ 0.05).Conclusion The glutamate transporters GLAST and GLT-1 show different expression patterns in astrocytes and microglia following SCI in rats,which may be correlated with the roles of different glial cells in repair of spinal cord injury.

Role of JAK2-STAT3 signaling pathway in sevoflurane postconditioning-induced inhibition of mPTP opening during myocardial ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the role of Janus kinase 2-signal transducer and activator of transcription 3 (JAK2-STAT3) signaling pathway in sevoflurane postconditioning-induced inhibition of mitochondrial permeability transition pore (mPTP) opening during myocardial ischemia-reperfusion (I/R)in rats.Methods Sixty pathogen-free healthy male Sprague-Dawley rats,weighing 250-300 g,were divided into 4 groups (n=15 each) using a random number table:I/R group,sevoflurane postconditioning group (group SP),AG-490 group (group AG) and sevoflurane postconditioning plus AG-490 group (group SP+AG).Myocardial I/R was induced by 30 min ligation of the left anterior descending branch of coronary artery followed by 120 min reperfusion.In group SP,2.8％ sevoflurane was inhaled for 15 min starting from 2 min before reperfusion.JAK2 inhibitor AG-490 3 mg/kg was intravenously injected at 10 min before reperfusion in group AG.In group SP+AG,AG-490 3 mg/kg was intravenously injected at 10 min before reperfusion,and 2.8％ sevoflurane was inhaled for 15 min starting from 2 min before reperfusion.At 15 min of reperfusion,5 rats were sacrificed and myocardial specimens were obtained for determination of the expression of JAK2,phosphorylated JAK2 (p-JAK2),STAT3 and phosphorylated STAT3 (p-STAT3)in myocardial tissues by Western blot.The ratios of p-JAK2 to JAK2 expression (p-JAK2/JAK2) and pSTAT3 to STAT3 expression (p-STAT3/STAT3) were calculated.Five rats were sacrificed at the end of reperfusion for measurement of myocardial infarct size.The left 5 rats were selected and sacrificed,myocardial specimens were obtained,and the opening of mPTP was detected by a calcein-cobalt quenching method.Results Compared with group I/R,the myocardial infarct size and mPTP opening were significantly decreased,and JAK2/p-JAK2 and STAT3/p-STAT3 were increased in group SP (P＜0.05),and no significant change was found in the parameters mentioned above in SP+AG and AG groups (P＞0.05).Compared with group SP,the myocardial infarct size was significantly enlarged,the extent of mnPTP opening was aggravated,and JAK2/p-JAK2 and STAT3/p-STAT3 were decreased in SP+AG and AG groups (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning inhibits the opening of mPTP during myocardial I/R is related to activation of JAK2-STAT3 signaling pathway in rats.

Multiple modalities PET/CT neuroimaging for the diagnosis of Parkinson's disease and multiple system atrophy: statistical parametric mapping analysis / 中华神经科杂志

Objective To investigate the topographic distributions of dopamine transporter (DAT),dopamine D2 receptor and glucose in Parkinson's disease (PD) and multiple system atrophy (MSA) using positron emission tomography/computed tomography (PET/CT) scanning and statistical parametric mapping (SPM) analysis.Methods Seventy subjects (39 PD patients,15 MSA patients and 16 normal controls) who came from People's Liberation Army General Hospital from September 2013 to November 2015 underwent DAT,D2 receptor and glucose brain PET/CT scans using 11 C-methyl-N-2-β-carbomethoxy-3-β-(4-fluorophenyl) tropane (11C-β-CFT),11C-raclopride and 18F-fluorodeoxyglucose (18 F-FDG) as radiotracers,respectively.The uptake patterns were analyzed using SPM software.Results Striatal DAT binding decreased in the putamen in PD patients compared with controls (Z =5.21-5.77,P =0.002-0.016).D2 receptor showed no significant differences.However,glucose uptake decreased in cingulate gyrus(Z =4.51-4.67,P =0.010-0.017).For MSA patients,both DAT and D2 receptor binding decreased in the putamen(Z =2.13-3.42,P =0.000-0.016).Glucose uptake decreased in the bilateral putamen,cerebellum and part of frontal temporal lobes (Z =1.86-3.75,P =0.000-0.032).Conclusion Multiple modalities PET/CT scans using the ligands 11 C-β-CFT,11C-raclopride,and 18F-FDG are valuable in diagnosis of MSA and differential diagnosis of MSA from PD.

Assessment of effects of 7,8-dihydroxyflavone on the striatum in normal cynomolgus monkeys with 99TCm-TRODAT-1 SPECT/CT imaging / 中华核医学与分子影像杂志

Objective To assess the effects of 7,8-dihydroxyflavone (7,8-DHF) on the striatum (ST) in normal cynomolgus monkeys using 99Tcm-TRODAT-1 imaging.Methods A total of six healthy female cynomolgus monkeys were included in this study.Three of them were fed with normal food (control group),and the other three were given oral administration of 7,8-DHF in addition to normal food (experimental group).The SPECT/CT imaging was performed at different time after 99Tcm-TRODAT-1 injection.The ROI of ST was drawn on images of 3 consecutive transverse slices that could be visualized best.The cerebellum (CB) was taken as the background reference area.The radioactivity uptake ratios of ST/CB at 1,3,4 and 5 h were calculated respectively.Paired-t test was used to analyze the data.Results ST radioactive uptake ratios showed continuing increase on the delay images.ST/CB uptake ratios of the control group at 1,3,4 and 5 h were 1.43±0.04,1.82±0.06,2.04±0.12,2.42±0.23,respectively,and those of the experimental group were 1.35±0.08,2.40±0.09,2.74±0.13 and 3.25±0.15 respectively.There was no significant difference between the two groups at 1 h (t =2.57,P＞0.05),while ST/CB uptake ratios of the experimental group at 3,4 and 5 h were significantly higher (t values:2.77,2.87 and 2.92,all P＜0.05).Conclusion 99Tcm-TRODAT-1 SPECT/CT imaging can be used to assess the DAT activation effect by 7,8-DHF on ST of cynomolgus monkeys.

Effect of SLC6A4 gene polymorphism on pain sensitivity in patients with lung cancer / 中华麻醉学杂志

Objective To evaluate the effect of SLC6A4 gene polymorphism on pain sensitivity in the patients with lung cancer. Methods A total of 248 patients with pulmonary malignant tumor served as lung cancer group and 104 healthy subjects in the Physical Examination Center of our hospital served as con-trol group. Patients with malignant pulmonary tumor in lung cancer group were further divided into 3 sub-groups according to pain score and the three step analgesic ladder recommended by WHO(opioids was used when visual analogue scale[VAS]score≥4 points): painless subgroup, mild pain subgroup and moder-ate-severe pain subgroup. The consumption of opioids(based on requirement for morphine)within 24 h af-ter pain relief(VAS score≤3 points)after treatment and VAS score before treatment were recorded. Ve-nous blood samples were collected, and the genotypes were analyzed by polymerase chain reaction-restric-tion fragment length polymorphism. Results There was no significant difference in genotype frequency or allele frequency at rs4795541 and rs3813034 sites between lung cancer group and control group and among the three subgroups(P>0.05). There was no significant difference in VAS score before treatment or re-quirement for morphine between patients of different genotypes and alleles(P>0.05). There was no signifi-cant difference in VAS score before treatment or requirement for morphine between lung cancer patients with medium and low expression of serotonin transporter in 5-HTT-linked polymorphic region. Conclusion SLC6A4 gene polymorphism exerts no effect on pain sensitivity in the patients with lung cancer.

Effect of etomidate post-conditioning on mPTP in rat cortical neurons subjected to oxygen-glucose deprivation and restoration: the relationship with Robo receptors / 中华麻醉学杂志

Objective To evaluate the role of etomidate post-conditioning on mitochondrial permeability transition pore (mPTP) in the rat cortical neurons subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with Robo receptors.Methods The cortical neurons obtained from Sprague-Dawley rats (＜ 24 h after birth) were cultured in vitro and seeded in 6-well plates (2 ml/well).The neurons were divided into 4 groups (n=24 each) using a random number table:control group (group C),OGD/R group,etomidate post-conditioning group (group E),and etomidate post-conditioning + Robo receptor blocker group (group ER).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h.In E and ER groups,etomidate was added to the culture medium with the final concentration of 6 μmol/L immediately after onset of O2-glucose supply.In group ER,Robo blocker RoboN was added to the culture medium with the final concentration of 1 μg/ml at 6 h before O2-glucose deprivation.The neuronal apoptosis was detected using Hoechst/PI double staining,the viability of neurons was measured by MTT assay,and the amount of lactic dehydrogenase (LDH) released was measured using colorimetric method.The mitochondria were extracted,and mitochondrial permeability transition pore (mPTP) opening was detected.Results Compared with group C,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in OGD/R,E and ER groups (P＜0.05).Compared with group OGD/R,the apoptosis rate,amount of LDH released,and mPTP opening were significantly decreased,and the cell survival rate was increased in group E,and the apoptosis and amount of LDH released were significantly decreased,and the cell survival rate was increased in group ER (P＜0.05).Compared with group E,the apoptosis rate,amount of LDH released,and mPTP opening were significantly increased,and the cell survival rate was decreased in group ER (P＜0.05).Conclusion Etomidate post-conditioning mitigates OGD/R-induced damage to the cortical neurons through activating Robo receptors and inhibiting mPTP opening in rats.

Role of ERK signaling pathway in inhibition of OGD/R-induced apoptosis in rat cortical neurons by sevoflurane: the relationship with mPTP / 中华麻醉学杂志

Objective To evaluate the role of extracellular signal-regulated kinase (ERK) signaling pathway in inhibition of oxygen-glucose deprivation and restoration (OGD/R)-induced apoptosis in rat cortical neurons by sevoflurane and the relationship with mitochondrial permeability transition pore (mPTP).Methods The rat cortical neurons were cultured in vitro and seeded in 6-well or 12-well culture plates.The neurons were randomly divided into 4 groups (n =18 each) using a random number table:control group (group C);OGD/R group (group O);sevoflurane group (group OS);sevoflurane + ERK1/2 inhibitor PD98059 group (group OSP).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h in group O.The neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OS.OGD/R was performed at 1 h after ERK1/2 inhibitor PD98059 30 μmol/L was added,and the neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OSP.At 24 h of restoration of O2-glucose supply,the expression of phosphorylated ERK1/2 (p-ERK1/2) in neurons was measured by Western blot,the neuronal apoptosis was detected using Annexin V-FITC/PI double staining combined with flow cytometry,and the opening of mPTP was determined through measuring the optical density at 540 nm.The apoptosis rate was calculated.Results Compared with group C,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly increased in group O (P＜0.05).Compared with group O,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly decreased in group OS (P＜0.05).Compared with group OS,the expression of p-ERK1/2 in neurons was significantly down-regulated,and the apoptosis rate and mPTP opening were significantly increased in group OSP (P＜0.05).Conclusion The mechanism by which sevoflurane inhibits OGD/R-induced apoptosis in rat cortical neurons is related to inhibition of mPTP opening after activation of ERK signaling pathway.