Recent progress on the effects of structured lipids and fluidity of plasma membrane on tumor metastasis / 药学学报

The lipid composition of cell plasma membranes of aggressive tumors is significantly altered from normal, affecting the membrane fluidity and function. Plasma membrane fluidity involves multiple steps in tumor invasion and metastasis, including cell movement, adhesion, lateral diffusion of membrane molecules, signal transduction, material exchange and so on. This review highlights the difference in plasma membrane lipid composition and fluidity between normal and cancer cells, as well as the correlation with the invasion and metastasis potential of cancer. We also point out that the proliferation, invasion and metastasis of tumors can be inhibited by improving membrane fluidity or interfering with the membrane structured lipid composition, this focusing more on changing the biophysical properties of cancer cell membranes, and providing a novel strategy that works for treatment of tumor metastasis.

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.

Establishment and clinical verification of brain template for analyzing 11C-CFT PET imaging data / 中华核医学与分子影像杂志

Objective:To establish standard spatial brain template and ROIs template of 11C-methyl- N-2β-carbomethoxy-3β-(4-fluorophenyl)tropane (CFT) PET images for automated quantitative analysis of dopamine transporter (DAT) distribution. Methods:From May 2014 to December 2015, 11C-CFT PET and MRI T 1 brain images of 16 healthy volunteers (3 males, 13 females; age (63.3±6.9) years) from Huashan Hospital, Fudan University were co-registered and smoothed using statistical parametric mapping(SPM)5 software based on MATLAB to create a standard spatial brain template. The ROIs template was established by ScAnVp procedures. These templates were clinically verified by using 11C-CFT PET images of 37 healthy volunteers (23 males, 14 females; age (61.7±7.1) years), 32 Parkinson′s disease (PD) patients (20 males, 12 females; age (61.1±5.4) years), 10 multiple system atrophy with predominant parkinsonism (MSA-P) patients (7 males, 3 females; age (60.8±7.1) years) and 10 progressive supranuclear palsy (PSP) patients (5 males, 5 females; age (58.4±6.1) years) from Huashan Hospital, Fudan University between January 2014 and March 2019. One-way analysis of variance was used to analyze data. Results:Based on the 11C-CFT PET images and MRI T 1 images of healthy volunteers, a standard spatial brain template for normalization of 11C-CFT PET images was created. The ROIs template was established including seven regions: bilateral caudate, anterior putamen, posterior putamen (along the long axis) and the occipital cortex. The ROIs template was accurately aligned in each verification group. The normal reference values of semi-quantitative DAT distribution in caudate, anterior putamen and posterior putamen were obtained (1.84±0.13, 2.18±0.16, 1.77±0.11). The semi-quantitative values of 11C-CFT uptake in each ROI in patients were significantly lower than those in healthy volunteers ( F values: 49.79-283.83, all P<0.05). Conclusion:The established brain templates with accurate spatial alignment for 11C-CFT image analysis can provide foundational tools for the application of 11C-CFT PET imaging in clinical practice and scientific research.

ABCA1 transporter promotes the motility of human melanoma cells by modulating their plasma membrane organization

BACKGROUND: Melanoma is one of the most aggressive and deadliest skin tumor. Cholesterol content in melanoma cells is elevated, and a portion of it accumulates into lipid rafts. Therefore, the plasma membrane cholesterol and its lateral organization might be directly linked with tumor development. ATP Binding Cassette A1 (ABCA1) transporter modulates physico-chemical properties of the plasma membrane by modifying cholesterol distribution. Several studies linked the activity of the transporter with a different outcome of tumor progression depending on which type. However, no direct link between human melanoma progression and ABCA1 activity has been reported yet. METHODS: An immunohistochemical study on the ABCA1 level in 110 patients-derived melanoma tumors was performed to investigate the potential association of the transporter with melanoma stage of progression and prognosis. Furthermore, proliferation, migration and invasion assays, extracellular-matrix degradation assay, immunochemistry on proteins involved in migration processes and a combination of biophysical microscopy analysis of the plasma membrane organization of Hs294T human melanoma wild type, control (scrambled), ABCA1 Knockout ( ABCA1 KO) and ABCA1 chemically inactivated cells were used to study the impact of ABCA1 activity on human melanoma metastasis processes. RESULTS: The immunohistochemical analysis of clinical samples showed that high level of ABCA1 transporter in human melanoma is associated with a poor prognosis. Depletion or inhibition ofABCA1 impacts invasion capacities of aggressive melanoma cells. Loss of ABCA1 activity partially prevented cellular motility by affecting active focal adhesions formation via blocking clustering of phosphorylated focal adhesion kinases and active integrin ß3. Moreover, ABCA1 activity regulated the lateral organization of the plasma membrane in melanoma cells. Disrupting this organization, by increasing the content of cholesterol, also blocked active focal adhesion formation. CONCLUSION: Human melanoma cells reorganize their plasma membrane cholesterol content and organization via ABCA1 activity to promote motility processes and aggressiveness potential. Therefore, ABCA1 may contribute to tumor progression and poor prognosis, suggesting ABCA1 to be a potential metastatic marker in melanoma.

Construction and optimization of a novel plasma membrane model for membrane affinity assay based on polydiacetylene vesicles / 药学学报

Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.

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.

Oocyte quality assessment in marine invertebrates: a novel approach by fluorescence spectroscopy

BACKGROUND: The assessment of oocyte quality is, nowadays, a major challenge in aquaculture, oocyte cryopreservation, and environmental science. Oocyte quality is a determining factor in fertilization and embryo development; however, there is still a lack of rapid and sensitive cellular markers for its assessment. Currently, its estimation is pre-dominantly based on morphological analysis, which is subjective and does not consistently reflect the developmental competence of the oocytes. Despite several recent studies investigating molecular markers related to oocyte quality, methods currently available for their determination pose various technical challenges and limitations. In this study, we developed a novel approach based on fluorescence spectroscopy to assess different intrinsic physiological parameters that can be employed to evaluate egg quality in marine invertebrates that are widely used as animal models such as sea urchins and mussels. RESULTS: Different physiological parameters, such as viability, mitochondrial activity, intracellular ROS levels, plasma membrane lipid peroxidation, and intracellular pH, for egg quality evaluation have been successfully assessed in sea urchins and mussels by using specific fluorescent dyes and detecting the fluorescent signals in eggs through fluorescence spectroscopy. CONCLUSIONS: Based on our findings, we propose these physiological markers as useful predictors of egg quality in marine invertebrates; they can be estimated rapidly, selectively, and sensitively by employing this novel approach, which, due to the speed of analysis, the low cost, and easy use can be considered a powerful analytical tool for the egg quality assessment.

Effect of Zuoguiwan on Expression of Ca2+ Transport Pathway-related Protein in Ovariectomized Rats / 中国实验方剂学杂志

Objective::To explore the effect of Zuoguiwan on the bone mineral density (BMD) and the expressions of Ca2+ transport-associated protein in ovariectomized rats. Method::The 48 female SD rats were randomly divided into six groups: normal group, model group, sham operation group, estrogen group(0.167 mg·kg-1) and low and high-dose Zuoguiwan groups(9.6, 38.4 g·kg-1), with 10 rats in each group. Except for the sham-operated group, the ovariectomized rats in the other groups received the bilateral ovariectomy. Therapeutic intervention was given in each group for 3 months after the establishment of the model. After 12 weeks, BMD was measured using dualenergy X-ray absorptiometry. Tartrated presistant acid phosphatse(TRACP) and serum calcium were detected by biochemical kits.Protein expression in Ca2+ transport (Bone tissue) was detected by Western blot. Result::Compared with the normal group, the serum calcium of the model group was decreased(P<0.01). Compared with the normal group, BMD of the model group was decreased (P<0.01). The serum calcium of rats in high-dose group and western medicine group was higher than that of model group(P<0.01). BMD in model group was lower than that of Zuoguiwan groups and estrogen group(P<0.05). There was no significant difference in TRACP among the groups. Nilestriol and Zuoguiwan can down-regulate the expressions of TRPV5, NCX1, CaBP-D28K and PMCA1b in bone tissue of castrated rats(P<0.05, P<0.01). Conclusion::Zuoguiwan can down-regulate the expressions of Ca2+ transport-associated proteins (Bone tissues) in rat osteoclasts, with an efficacy on osteoporosis.

CtpB is a plasma membrane copper (I) transporting P-type ATPase of Mycobacterium tuberculosis

BACKGROUND: The intracellular concentration of heavy-metal cations, such as copper, nickel, and zinc is pivotal for the mycobacterial response to the hostile environment inside macrophages. To date, copper transport mediated by P-type ATPases across the mycobacterial plasma membrane has not been sufficiently explored. RESULTS: In this work, the ATPase activity of the putative Mycobacterium tuberculosis P1B-type ATPase CtpB was associated with copper (I) transport from mycobacterial cells. Although CtpB heterologously expressed in M. smegmatis induced tolerance to toxic concentrations of Cu2+ and a metal preference for Cu+, the disruption of ctpB in M. tuberculosis cells did not promote impaired cell growth or heavy-metal accumulation in whole mutant cells in cultures under high doses of copper. In addition, the Cu+ ATPase activity of CtpB embedded in the plasma mem-brane showed features of high affinity/slow turnover ATPases, with enzymatic parametersKM 0.19 ± 0.04 µM and Vmax 2.29 ± 0.10 nmol/mg min. In contrast, the ctpB gene transcription was activated in cells under culture conditions that mimicked the hostile intraphagosomal environment, such as hypoxia, nitrosative and oxidative stress, but not under high doses of copper. CONCLUSIONS: The overall results suggest that M. tuberculosis CtpB is associated with Cu+ transport from mycobacterial cells possibly playing a role different from copper detoxification.

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)

Papain-mediated reduction of MTT is an artifact but not an indicator of cell apoptosis in Saccharomyces cerevisiae

Papain is a proteolytic enzyme of the cysteine protease family used for tissue dissociation and cell separation. Papain’snonspecific proteolysis of the plasma membrane enzymes plays a crucial role in the homeostasis by disrupting theintracellular pH of the affected cells which might lead to cell death. When the Saccharomyces cerevisiae cellswere treated with different concentrations of papain (1.0, 5.0, 10.0, 20.0, and 30.0 µg/ml), we found no alterationin the trans-plasma membrane electron transport (TPMET) activity and the intracellular pH of the cells, while itsignificantly decreased the mitochondrial dehydrogenase activity when measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. Additional verification of cell viability by trypan blue assay showed98%–99% cell viability, contrary to the higher cell death observed with MTT assay. To better understand the decreasein cell viability with MTT assay, we tested the cell-free system that demonstrated a significant decrease of MTTconcentration but the trypan blue assay showed more number of viable cells. This study shows that papain interfereswith the MTT assay.

Cell Death and Its Different Modes: History of Understanding and Current Trends

Discussions about what is life continue to struggle; there are pros and cons for whether a virus is alive. However, an opposite thing –cell death –appears to be tantamount important and equally not-easygoing to define. Nevertheless, our current knowledgeabout eukaryotic cell death has made a long way and resulted in a fruitful outcome: starting from three types of cell death (type I, II and III which are mainly applicable to eukaryotic cells of organisms from the biological kingdom animalia) in 1970s, Nomenclature Committee on Cell Death has named already twelve cell death forms in 2018, including the above mentioned apoptosis, autophagy and necrosis among them. How the scientific attitude towards cellular demise evolved and various aspects of different cell death modes are reviewed in this article.

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.

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.

Colesterol carregado pela ciclodextrina sobre a criopreservação dos espermatozoides de garanhões da raça Nordestina / Cholesterol-loaded cyclodextrin on the cryopreservation of stallion sperm of the Northeastern breed

Nesta pesquisa avaliou-se o efeito do colesterol sobre o sêmen de garanhões da raça Nordestina sobre a qualidade espermática. Vinte ejaculados de dois garanhões foram diluídos com BotuSemen e colesterol carreado pela ciclodextrina (CCC) adicionado no sêmen: controle, 0,75mg de CCC e 1,0mg de CCC/120x106 sptz/mL, e incubado a 26°C/15min. O sêmen foi diluído 1:5 (v/v) com diluente Lactose-gema de ovo e resfriado a 5°C/2h, envasado em palhetas de 0,5mL, e acondicionado sob vapor de nitrogênio líquido, e depois imersos. As amostras foram descongeladas (37 °C/30s) e avaliadas. As variáveis foram avaliadas com ANOVA e teste de Tukey (P<0,05). A motilidade total e progressiva foi maior (P<0,05) no sêmen tratado com CCC comparado as amostras do grupo controle, e CCC promoveu maior percentual (P<0,05) de motilidade total e progressiva durante as 3 horas de incubação. A percentagem de espermatozoides com viabilidade e integridade foi maior (P<0,05) no sêmen tratado com CCC (81,47 e 86,07%) comparado ao controle (72,12 e 70,19%). O número de espermatozoides reativos ao teste hiposmótico foi maior (P<0,05) nas amostras de sêmen tratadas com CCC comparado ao controle. Adição de colesterol no sêmen de garanhões Nordestino melhora a qualidade espermática apos a criopreservação.(AU)

In the study effect of cholesterol was evaluated on the sperm quality of Nordestina stallion breed. Twenty semen samples were used from two stallions, diluted with BotuSemen extender and cholesterol add as follows: control, 0.75mg of cholesterol-loaded cyclodextrin (CLC) and 1.0mg of CLC/120x106 sperm/mL, and incubated for 15 min at 22°C. The samples were diluted 1:5 with lactose-yolk egg extender and cooled to 5°C over two hours, loaded into 0.5mL straws and frozen in static liquid nitrogen vapor before being plunged into nitrogen. Samples were thawed (37°C/30s) and analyzed. The variables were analyzed by ANOVA and means compared by Tukey test (P<0.05). Higher percentages of total and progressive motile was higher for sperm treated with CLC compared to control, and CLC promoted higher percentages (P<0.05) of total and progressive motility for the 3 hours of incubation. The percentage of viability and plasma membrane integrity of spermatozoa were higher (P<0.05) in sperm treated with CLC compared to the control group. The number of spermatozoa reacted to hypoosmotic test was higher (P<0.05) in sperm treated with CLC than control. Addition of CLC in the semen of Nordestina stallion breed improve the sperm quality after cryopreservation.(AU)

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.

Effect of plasma membrane monoamine transporter genetic variants on pharmacokinetics of metformin in humans

Metformin, an oral hypoglycemic agent belonging to biguanide class, is widely used to treat type 2 diabetes mellitus, and several drug transporters such as organic cation transporters (OCTs), multidrug and toxin extrusion transporter (MATE), and plasma membrane monoamine transporter (PMAT) are thought to affect its disposition. We evaluated the role of PMAT genetic variations on the pharmacokinetic characteristics of metformin in a Korean population. In this retrospective study, 91 healthy subjects from four different metformin pharmacokinetic studies were analyzed; in each study, the subjects were administered two oral doses of metformin at intervals of 12 hours and dose-normalized pharmacokinetic parameters were compared between the subjects' genotypes. Subjects who had more than one allele of c.883-144A>G single nucleotide polymorphism (SNP) in PMAT gene (rs3889348) showed increased renal clearance of metformin compared to wild-type subjects (814.79 ± 391.73 vs. 619.90 ± 195.43 mL/min, p=0.003), whereas no differences in metformin exposure were observed between the PMAT variant subjects and wild-type subjects. Similarly, subjects with variant rs316019 SNP in OCT2 showed decreased renal clearance of metformin compared to wild-type subjects (586.01 ± 160.54 vs. 699.13 ± 291.40 mL/min, p=0.048). Other SNPs in PMAT and MATE1/2-K genes did not significantly affect metformin pharmacokinetics. In conclusion, the genetic variation of c.883-144A>G SNP in PMAT significantly affects the renal clearance of metformin in healthy Korean male subjects.

Peeling the onion: the outer layers of Cryptococcus neoformans

Cryptococcus neoformans is an opportunistic fungal pathogen that is ubiquitous in the environment. It causes a deadly meningitis that is responsible for over 180,000 deaths worldwide each year, including 15% of all AIDS-related deaths. The high mortality rates for this infection, even with treatment, suggest a need for improved therapy. Unique characteristics of C. neoformans may suggest directions for drug discovery. These include features of three structures that surround the cell: the plasma membrane, the cell wall around it, and the outermost polysaccharide capsule. We review current knowledge of the fundamental biology of these fascinating structures and highlight open questions in the field, with the goal of stimulating further investigation that will advance basic knowledge and human health.

Effect of spinal cord stimulation on expression of spinal GLT-1 and GLAST in rats with diabetic neu-ropatbic pain / 临床麻醉学杂志

Objective To observe the effect of spinal cord stimulation on expression of spinal GLT-1 and GLAST in rats with diabetic neuropatbic pain.Methods Forty-eight healthy male SD rats,only 2 months old,weighting 250-300 g,using the random number table method,were divided into four groups (n=12):the control group (group C),diabetes neuralgia group (group D),false stimulation group (group N)and spinal cord stimulation group (group S).The model of diabetes was induced by the pedtoneal injec-tion of streptozocin (STZ),electrodes were placed into the epidural space 1 9 days after injection of STZ in groups N and S,in addition,group S was performed 26-28 days after injection of STZ.Mechanical contrac-tion leg threshold (MWT)were determined one day before STZ injection,2 d,7 d,14 d and 28 d after STZ injection.The rats were sacrificed,the lumbar spinal cord tissue were obtained for determination of GLT-1 and GLAST expression in spinal cord tissues on 28 d after measurement of MWT.Results Compared with group C,MWT was decreased 14 d and 28 d after STZ injection,and expression levels of GLT-1 and GLAST mRNA were increased on 14 d and 28 d (P<0.05);Compared with before STZ injection,MWT of group D,group N and group S was decreased on 14 d and 28 d (P<0.05);Compared with group D, MWT was increased,and expression levels of GLT-1 and GLAST mRNA were increased in group S on 28 d after STZ injection (P<0.05 ).Conclusion The mechanism of spinal cord stimulation reducing rats diabetes neuralgia may be related to elevating the expression of GLT-1 and GLAST.