Targeted lipidomics uncovers oxylipin perturbations and potential circulation biomarkers in Bietti's crystalline dystrophy.

PURPOSE: Abnormalities in lipid metabolism have been proposed in Bietti's crystalline dystrophy (BCD). We aim to characterize the lipid profiles in a case-control study. METHODS: All participants were genetically confirmed by CYP4V2 gene sequencing and underwent chorioretinopathy evaluation by calculating the percentages of AF atrophy (PAFA). Fasting blood samples of BCD patients and controls were collected, and plasma was analyzed for routine lipid profiles. Targeted lipidomic evaluation includes long chain polyunsaturated fatty acids (LCPUFA) and associated eicosanoid metabolites. RESULTS: Routine lipids profiles showed elevated plasma levels of triglyceride (P = 0.043) and low-density lipoprotein cholesterol (P = 0.024) in BCD patients. Lipidomic analysis showed significantly decreased levels of ω-3 LCPUFA including docosahexaenoic acid (DHA, 22:6, P = 0.00068) and eicosapentaenoic acid (EPA, 20:5, P = 0.0016), as well as ω-6 LCPUFA arachidonic acid (ARA, 20:4, P < 0.0001) in BCD patients. Eicosanoid metabolites, either derived from ω-3 and/ or ω-6 LCPUFAs via cyclooxygenase (COX) or lipoxygenase (LOX) pathways, including 5-HEPE, 12-HEPE, 13-HDHA, 15-HETE, 12-HETE, 5-HETE, 6k-PGF1a, PGE2, PGJ2, and TXB2, exhibited significant differences (P < 0.0001) between BCD patients and controls. Genotypes of CYP4V2, specifically the biallelic null mutations, were observed to correlate with more remarkably reduced levels of oxylipins, involving major LOX pathway metabolites including 5-HETE, 5-HEPE, 12-HEPE and LTB4. CONCLUSIONS: BCD patients demonstrated significant decreases in plasma levels of ω-3 and ω-6 LCPUFA (DHA, EPA, and ARA), as well as their downstream metabolites via the COX and LOX pathways, suggesting that these might be implicated in BCD pathogenesis and could serve as biomarkers and therapeutic targets of the disease. KEY MESSAGES: What is known BCD is a vision-threatening hereditary disease the causative gene of which is CYP4V2. Abnormalities in lipid metabolism have been proposed and demonstrated previously in BCD studies. The detailed pathogenesis remains unclear and controversial. What is new We observed prominent lipidomic alterations in the circulation when compared with age, gender, and bodymass index (BMI)-matched healthy controls. BCD patients demonstrated significant decreases in plasma levels of ω-3 and ω-6 LCPUFA (DHA, EPA, and ARA). Remarkable changes were observed in the downstream metabolites of the LCPUFA via the COX and LOX pathways. Genotypes of CYP4V2, specifically the biallelic null mutations, were observed to correlate with more remarkably reduced levels of oxylipins, involving major LOX pathway metabolites.

Contribution of differential alteration in oxidative stress and anti-oxidation related molecular signals to toxicity difference between atrazine and its main metabolites in nematodes.

As a widely used herbicide, atrazine and its two main metabolites of deethylatrazine (DEA) and deisopropylatrazine (DIA) pose an exposure risk for both human beings and animals in the environment. In this study, Caenorhabditis elegans was selected as an in vivo model to compare the toxicity between atrazine and its main metabolites. Upon exposure from the larval stage L1 to adult day 3, both DEA and DIA showed less toxicity on locomotion and reproduction compared with atrazine at concentration of 0.001, 0.01 0.1 and 1 mg/L for parental generation. In addition, exposure to DEA and DIA at concentration of 0.1 mg/L also induced less transgenerational toxicity on locomotion than exposure to atrazine for both parental generation and offspring of F1-F4. Accordingly, exposure to DEA and DIA caused less ROS production and alteration in the expression of some genes (mev-1, gas-1, and clk-1) governing oxidative stress compared to atrazine. Meanwhile, DEA and DIA lead to less increase in expression of superoxide dismutase genes (sod-2 and sod-3) and SOD-3::GFP than atrazine. Moreover, atrazine and its two main metabolites differentially activated the daf-16 encoding FOXO transcriptional factor in insulin signaling pathway during the control of downstream target of SOD-3. Overall, our results highlighted the important role of oxidative stress and anti-oxidation related molecular signals in mediating toxicity of atrazine, DEA and DIA, which provided a novel explanation for the different toxicity between atrazine and its main metabolites.

Proteomics Analysis Identifies IRSp53 and Fascin as Critical for PRV Egress and Direct Cell-Cell Transmission.

Pseudorabies virus (PRV) has been widely used as a live trans-synaptic tracer for mapping neuronal circuits. Systematically identifying mature PRV virion proteomes and defining co-purified host proteins are necessary to fully understand the detailed mechanism underlying PRV transmission processes. Here, a PRV virion purification strategy based on sorting with flow cytometry is developed and the mature extracellular and intracellular PRV virion proteomes using LC coupled with MS/MS are characterized. In addition to viral proteins, a large number of host proteins are also identified, including proteins related to actin cytoskeletal dynamics and membrane protrusion. How many of these host proteins are true virion components are unknown and the majority of these may not be. Through functional analysis, it is found that IRSp53 and fascin are critical for the egress process and play a role in direct cell-cell transmission. Moreover, it is shown that CDC42 and Rac1 are also involved in the production of mature extracellular virions. The results suggest that the formation of the filopodia-like cytoskeleton and the rearrangement of the membrane, which are both associated with IRSp53 and fascin, may be important for the transmission of viruses used in neuronal tracing.

Decreased Network Efficiency in Benign Epilepsy with Centrotemporal Spikes.

Purpose To investigate the functional connectome alterations in benign epilepsy with centrotemporal spikes with respect to the occurrence of interictal epileptic discharges (IEDs) during functional magnetic resonance (MR) imaging. Materials and Methods This prospective study was approved by the local institutional review board and was HIPAA compliant. All participants were consecutively enrolled with written informed consent. Forty-three right-handed patients were classified into IED (n = 20, 13 girls and seven boys; mean age ± standard deviation, 9.00 years ± 1.95) and non-IED (n = 23, 11 girls and 12 boys; mean age, 10.22 years ± 2.13) groups on the basis of electroencephalographic data simultaneously recorded during resting-state functional MR imaging at 3.0 T. The functional connectome features (estimated with graph theoretical analysis) in patient groups and control subjects who were matched for sex, age, and education level (n = 28, all right-handed, 13 girls and 15 boys; mean age, 10.00 years ± 2.31) were compared by using one-way analysis of variance. Results Patients with IEDs and those without IEDs showed consistently abnormal global topology in their functional networks (ie, decreased global efficiency; P < .05) relative to that of control subjects, with no differences between the two patient groups (P > .05). Decreased regional efficiency and connectivity strength were observed in the patients with IEDs and those without (mainly in the perirolandic and frontal areas) relative to control subjects (P < .05). Moreover, the altered functional features significantly correlated with clinical characteristics (ie, disease duration and age at symptom onset, P < .05). Conclusion These findings suggest that decreased global and regional efficiency are prominent functional deficits in children with benign epilepsy with centrotemporal spikes and can be readily identified with resting-state functional MR imaging, irrespective of IEDs. © RSNA, 2016 Online supplemental material is available for this article.

Pseudo-typed Semliki Forest virus delivers EGFP into neurons.

Semliki Forest virus (SFV), a neurotropic virus, has been used to deliver heterologous genes into cells in vitro and in vivo. In this study, we constructed a reporter SFV4-FL-EGFP and found that it can deliver EGFP into neurons located at the injection site without disseminating throughout the brain. Lacking of the capsid gene of SFV4-FL-EGFP does not block its life cycle, while forming replication-competent virus-like particles (VLPs). These VLPs hold subviral genome by using the packaging sequence (PS) located within the nsP2 gene, and can transfer their genome into cells. In addition, we found that the G protein of vesicular stomatitis virus (VSVG) can package SFV subviral genome, which is consistent with the previous reports. The G protein of rabies virus (RVG) could also package SFV subviral genome. These pseudo-typed SFV can deliver EGFP gene into neurons. Taken together, these findings may be used to construct various SFV-based delivery systems for virological studies, gene therapy, and neural circuit labeling.

Thalamocortical dysconnectivity in paroxysmal kinesigenic dyskinesia: Combining functional magnetic resonance imaging and diffusion tensor imaging.

BACKGROUND: Paroxysmal kinesigenic dyskinesia is associated with macrostructural and microstructural abnormalities in the thalamus. OBJECTIVES: To examine functional and structural connectivity of thalamocortical networks in paroxysmal kinesigenic dyskinesia and to further investigate the effect of mutation of the proline-rich transmembrane protein 2 on thalamocortical networks. METHODS: Patients with paroxysmal kinesigenic dyskinesia (n = 20), subdivided into proline-rich transmembrane protein 2-mutated (n = 8) and nonmutated patients (n = 12) and healthy controls (n = 20) underwent resting-state functional MRI and diffusion imaging scan. The functional properties of correlations in neural activity (functional connectivity) and the structural properties of white matter probabilistic tractography (structural connectivity) were analyzed to characterize thalamocortical networks. Furthermore, the effect of proline-rich transmembrane protein 2 mutation on functional and structural connectivity of thalamocortical networks were examined using one-way analysis of variance among three groups. RESULTS: Patients had increased functional and structural connectivity between ventral lateral/anterior thalamic nuclei and a lateral motor area, as compared to controls. This functional connectivity positively correlated with disease duration. Interestingly, proline-rich transmembrane protein 2-mutated patients showed decreased functional connectivity and preserved structural connectivity, between mediodorsal nucleus and prefrontal cortex, compared to nonmutated patients and controls. CONCLUSIONS: Thalamomotor/premotor hyperconnectivity suggests abnormal communication between thalamus and motor cortex in patients. Furthermore, thalamoprefrontal hypoconnectivity in proline-rich transmembrane protein 2-mutated patients might indicate that proline-rich transmembrane protein 2 mutations result in inefficient thalamoprefrontal integration. Our findings facilitate a deeper understanding of the crucial role of thalamocortical dysconnectivity in the pathophysiological mechanisms of paroxysmal kinesigenic dyskinesia. © 2017 International Parkinson and Movement Disorder Society.

[Value of percentage of highly fluorescent lymphocytic cells for rapidly assessing septicemia in tumor patients].

OBJECTIVE: To evaluate the value of percentage of highly fluorescent lymphocytic cells (HFLC%) for rapidly assessing septicemia in tumor patients. METHODS: Blood samples were collected from 130 patients with tumors (60 septicemia patients and 70 non-septicemia patients) and 80 healthy controls. HFLC% was analyzed with Sysmex XE-5000, the level of C-reactive protein (CRP) measured with a commercially available turbidimetric immunoassay kit and the level of procalcitonin (PCT) determined with a semiquantitative chromatographic immunoassay kit. The diagnostic values of HFLC% and CRP in septicemia were evaluated with ROC analysis. RESULTS: The values of HFLC% and CRP were significantly higher in the septicemia group than those in the non-septicemia and healthy groups (0.30% (0.10%-0.70%) vs 0.10% (0-0.20%), 0.10% (0-0.20%) ; 80.3 (28.5-129.5) vs 3.3 (1.4-41.4) , 1.4 (0.6-2.5) mg/L, all P < 0.01) . The ROC-AUCs for HFLC% and CRP for a diagnosis of septicemia were 0.72 (sensitivity 71.7%, specificity 58.7%) and 0.92 (sensitivity 96.7%, specificity 82.0%). Both of them could judge septicemia better. Additionally, HFLC% was correlated with the levels of PCT and CRP (r = 0.637, 0.241, both P < 0.01). CONCLUSIONS: HFLC% may be used as a rapid and simple auxiliary indicator in the diagnosis of septicemia in patients with tumors. And it is conducive to make an early diagnosis of septicemia and avoid unnecessary use of antibiotics.

Erratum to "Lipidome Atlas of the Developing Heart Uncovers Dynamic Membrane Lipid Attributes Underlying Cardiac Structural and Metabolic Maturation".

[This corrects the article DOI: 10.34133/research.0006.].

[Comparison of testing methods for reticulated platelets].

OBJECTIVE: To compare the accuracy, stability and sample cross-contamination of two independent methods of detecting the percentage of reticulated platelets in peripheral blood and establish a local normal reference range so as to provide methodological rationales in clinical laboratory. METHODS: The percentages of reticulated platelets in peripheral blood of a healthy population were measured by Sysmex XE-5000 blood cell analyzer with polymethyl oxazine staining and flow cytometer with thiazole orange staining respectively. The correlation between the results of two methods was analyzed by Spearman's nonparametric correlation. Information about stability was obtained from measurements of the percentages of reticulated platelets in peripheral blood at designated time points. The analyses of accuracy, sample cross contamination and local normal reference range were performed routinely. RESULTS: The coefficient of variation (CV) of data was lower (16.2%) than that from flow cytometer (35.1%). The sample cross-contaminations of two methods were the same at around 5%. The percentage of reticulated platelets in peripheral blood was stable and consistent whereas the results of flow cytometer fluctuated at different time points within 4 h after blood sampling. The correlation of results obtained from two methods was significant (P < 0.01, r(2) = 0.923). The local normal reference range was 1.0% - 7.5% for Sysmex XE-5000 versus 3.0% - 10.5% for flow cytometer. CONCLUSIONS: Fully automatic blood cell analyze is more advanced than flow cytometer for its simple operation and stable data. And the former is an ideal first-choice for detecting the percentage of reticulated platelets in peripheral blood.

Involvement of indoleamine 2,3-dioxygenase in impairing tumor-infiltrating CD8 T-cell functions in esophageal squamous cell carcinoma.

The indoleamine 2,3-dioxygenase-(IDO-) mediated microenvironment plays an important role in tumor immune escape. However, the inhibitory effects of IDO on the CD8(+) tumour-infiltrating lymphocytes (CD8(+) TILs) in esophageal squamous cell carcinoma (ESCC) have not been clarified yet. Here, we found that the level of IDO expression in ESCC tumor specimens correlated with a reduction in the number of CD8(+) TILs. Patients with high IDO expression and a low number of CD8(+) TILs had significantly impaired overall survival time. IDO expression and functional enzyme activity in ESCC cell lines could be induced by IFNγ. When exposed to the milieu generated by IDO-expressing Eca109 cells, the CD8(+) TILs were suppressed in proliferation, and their cytolytic functions against target tumor cells were lost. These results suggested that impairing CD8(+) TIL functions by IDO expressed in ESCC possibly contributed to the finding that patients with higher IDO expression have more aggressive disease progression and shorter overall survival time.

Geochemistry and Organic Petrology of Middle Permian Source Rocks in Taibei Sag, Turpan-Hami Basin, China: Implication for Organic Matter Enrichment.

The Taodonggou group of Middle Permian is an important source rock in Taibei sag of Turpan-Hami basin. Due to its deep burial, drilling has only been revealed in recent years. Based on organic petrology and organic geochemistry experiments, this paper studies the organic petrology, organic geochemistry, sedimentary environment, and hydrocarbon generation potential of source rocks in Taibei sag, Turpan-Hami basin, and reveals the influence of the sedimentary environment on the organic matter abundance of source rocks. The results are as follows: (1) The organic matter of the Middle Permian source rocks in Taibei sag of Turpan-Hami basin is mainly sapropelite and exinite. The vitrinite is mainly vitrodetrinite, and the exinite is mainly lamalginiite. (2) The total organic carbon content value is 0.55-6.08 wt %, and the average value is 2.58 wt %. The PG value ranges from 0.78 mg HC/g to 30.86 mg HC/g, and the average value is 4.88 mg HC/g. Chloroform asphalt "A" is 0.046-0.8767 wt %, and the average value is 0.285 wt %. The types of organic matter are mainly III and II-III, and the R o value is 0.628-1.49 wt % (average = 0.988 wt %). The T max distribution is 329-465 °C. The average temperature is 434.7 °C, which is in the mature stage (oil window stage). The Middle Permian source rocks are mainly very good to excellent source rocks with a good hydrocarbon generation potential. (3) The source rocks are deposited in a semihumid and semiarid climate. Organic matter is input as a mixed source. The early and late stages is dominated by terrestrial higher plants. The middle stage is dominated by lower aquatic organisms, and the sedimentary environment consists of weak reduction and weak oxidation environments. (4) In the study area, the abundance of organic matter has a weak negative correlation with CPI and a positive correlation with Pr/Ph and ∑C21-/∑C22+. Under the coaction of paleoclimate, organic matter input, and redox environment, the enrichment model of organic matter with high productivity and weak oxidation environment characteristics can also form excellent source rocks. This study is of great significance and provides theoretical guidance for the exploration of deep oil and gas resources.

[Correlation factors of ethylenediaminetetraacetic acid-dependent pseudothrombocytopenia in cancer patients].

OBJECTIVE: To investigate the correlation factors of ethylenediaminetetraacetic acid-dependent pseudothrombocytopenia (EDTA-PTCP) in cancer patients. METHODS: The potential correlation factors of EDTA-PTCP such as gender, age, case history, tumor types, therapeutic drugs and duration of EDTA-PTCP from cancer patients were analyzed based on the patient records from October 2007 to September 2009 at our cancer center. RESULTS: A total of 49 EDTA-PTCP cases from a pool of 55 000 cancer patients were collected. No correlation was found with gender (male 49.0%, female 51.0%), concurrent hypertension (20.4%)/diabetes (10.2%) or cancer types (1 - 11 cases each type). EDTA-PTCP appeared at pre-therapy (n = 13) and post-therapy (n = 36). Eleven cases (30.6%) were chemotherapy, 5 cases (13.9%) were radiotherapy plus chemotherapy, 15 case (41.7%) were tumor resection, 5 cases (13.9%) were interventional therapy in 36 patients whose EDTA-PTCP appeared post-therapy. The most frequency use in chemotherapy patients was dexamethasone (87.5%, 14/16), and in surgery patients was penicillin antibiotics (75.0%, 15/20). And its frequency was once (n = 18) and more than twice (n = 31). If the subjects were divided into 2 groups of non-treatment plus surgery and chemotherapy plus intervention on the basis of treatment course, there was a significant difference between two groups in proportion of patients whose duration of EDTA-PTCP ≤ 2 weeks (89.3% vs 47.6%, χ(2) = 10.22, P < 0.01). CONCLUSIONS: The incidence of EDTA-PTCP in cancer patients may be associated with therapeutic drugs, but not probably with gender, concurrent hypertension/diabetes, tumor types or therapeutic regimens. Duration of EDTA-PTCP may be associated with the treatment course.

Comparison of Supersonic Shear Wave Imaging-Derived Renal Parenchyma Stiffness Between Diabetes Mellitus Patients With and Without Diabetic Kidney Disease.

This study aims to evaluate the difference in renal parenchyma stiffness assessed by measuring Young's modulus (YM) using a supersonic shear wave imaging (SSI) technique among healthy patients and patients with type 2 diabetes mellitus (DM) with and without diabetic kidney disease (DKD). We analyzed the correlations of YM with clinical information and conventional ultrasound parameters. All patients (N = 124) were divided into three groups: (i) healthy patients (patients without kidney disease or type 2 DM, N = 31); (ii) patients with type 2 DM without kidney disease (N = 38); and (iii) patients with DKD (N = 55). Conventional and SSI ultrasound examinations were performed in all individuals for both kidneys. Then, we recorded renal length, width, parenchyma thickness, interlobar arterial resistive index (RI) and values of mean, mininum and maximum YM. The mean values of these parameters for the left and right kidneys were calculated for statistical analysis. Statistical significance was considered at p < 0.05. Among all ultrasound parameters, the mean YM demonstrated the largest area under the receiver operating characteristic (ROC) curve (0.860). The areas under the ROC curve (AUCs) for renal length, width, parenchyma thickness, interlobar arterial RI, minimum YM and maximum YM were 0.493, 0.616, 0.507, 0.733, 0.848 and 0.794, respectively. The corresponding cutoff value of mean YM was 31.73 kPa, with a sensitivity of 85.5% and a specificity of 71.0%. The mean YM in patients with type 2 DM without kidney disease (31.44 ± 3.83 kPa) was significantly higher than that in the healthy group (26.45 ± 4.32 kPa) and lower than that in the DKD group (37.60 ± 6.56 kPa). Patients with type 2 DM without kidney disease were considered as stage 0 of DKD. Thus, the mean YM in the control group was significantly lower than that in the stage 0, 2, 3, 4 and 5 subgroups. The mean YM in the stage 0-2 subgroups was lower than that in the stage 5 group, and the mean YM in the stage 0 group was lower than that in the stage 4 group. In the DKD group, the mean YM had a positive correlation with cystine-c (r = 0.634), urea (r = 0.596), creatine (r = 0.690), uric acid (r = 0.263), albumin/creatinine ratio (r = 0.428) and the presence or absence of diabetic retinopathy (r = 0.354). The mean YM also had a negative correlation with the estimated glomerular filtration rate (r = -0.657). SSI is a non-invasive method with which to diagnose DKD and has a performance superior to that of conventional ultrasound. In addition, SSI may provide a secondary index for the staging of DKD and the monitoring of renal damage in type 2 DM patients.

Optimization of the Fluorescent Protein Expression Level Based on Pseudorabies Virus Bartha Strain for Neural Circuit Tracing.

Mapping the neural circuits facilitates understanding the brain's working mechanism. Pseudorabies virus (PRV; Bartha stain) as a tracer can infect neurons and retrogradely transport in neural circuits. To illuminate the network, tracers expressing reporter genes at a high level are needed. In this study, we optimized the expression level of reporter genes and constructed two new retrograde trans-multisynaptic tracers PRV531 and PRV724, which separately express more robust green and red fluorescent proteins than the existing retrograde tracers PRV152 and PRV614. PRV531 and PRV724 can be used for mapping the neural circuit of the central nervous system (CNS) and the peripheral nervous system (PNS). Overall, our work adds two valuable tracers to the toolbox for mapping neural circuits.

An integrated method for direct interrogation of sphingolipid homeostasis in the heart and brain tissues of mice through postnatal development up to reproductive senescence.

Development of rapid metabolomic methods poised for pathway discovery is expected to facilitate the identification of therapeutic candidates in the metabolomic approach to translational medicine. Using sphingolipid homeostasis as a prototype, we present herein an integrated method to facilitate a fast interrogation of altered sphingolipid (and phospholipid) metabolism associated with perturbed endolysosomal functions in mammalian systems. Constructed upon high performance liquid chromatography coupled to mass spectrometry, this method allows semi-quantitative measurements of more than 300 individual species within 20 min. The method was applied to investigate cardiac- and neural-specific developmental changes in sphingolipid regulation from the postnatal stage to reproductive senescence in mice, revealing that endogenous lysobisphosphatidic acids and specific complex glycosphingolipids are tightly co-regulated to foster concerted reductions in sphingolipid levels at distinct stages of postnatal development. Our lipidomic data suggest that such changing regulatory patterns in sphingolipid homeostasis is attributed to differential endolysosomal degradation of complex sphingolipids, which may be critical in ensuring efficient sphingolipid catabolism and organismal health at each stage of postnatal development.

Inter-hemispheric Intrinsic Connectivity as a Neuromarker for the Diagnosis of Boys with Tourette Syndrome.

Tourette syndrome (TS) is associated with gross morphological changes in the corpus callosum, suggesting deficits in inter-hemispheric coordination. The present study sought to identify changes in inter-hemispheric functional and anatomical connectivity in boys with "pure" TS as well as their potential value for clinical diagnosis. TS boys without comorbidity (pure TS, n = 24) were selected from a large dataset and compared to age- and education-matched controls (n = 32). Intrinsic functional connectivity (iFC) between bilateral homotopic voxels was computed and compared between groups. Abnormal iFC was found in the bilateral prefronto-striatum-midbrain networks as well as bilateral sensorimotor and temporal cortices. The iFC between the bilateral anterior cingulate cortex (ACC) was negatively correlated with symptom severity. Anatomical connectivity strengths between functionally abnormal regions were estimated by diffusion probabilistic tractography, but no significant between-group difference was found. To test the clinical applicability of these neuroimaging findings, multivariate pattern analysis was used to develop a classification model in half of the total sample. The classification model exhibited excellent classification power for discriminating TS patients from controls in the other half samples. In summary, our findings emphasize the role of inter-hemispheric communication deficits in the pathophysiology of TS and suggest that iFC is a potential quantitative neuromarker for clinical diagnosis.

Frequency-Specific Local Synchronization Changes in Paroxysmal Kinesigenic Dyskinesia.

The neurobiological basis of paroxysmal kinesigenic dyskinesia (PKD) is poorly defined due to the lack of reliable neuroimaging differences that can distinguish PKD with dystonia (PKD-D) from PKD with chorea (PKD-C). Consequently, diagnosis of PKD remains largely based on the clinical phenotype. Understanding the pathophysiology of PKD may facilitate discrimination between PKD-D and PKD-C, potentially contributing to more accurate diagnosis. We conducted resting-state functional magnetic resonance imaging on patients with PKD-D (n = 22), PKD-C (n = 10), and healthy controls (n = 32). Local synchronization was measured in all 3 groups via regional homogeneity (ReHo) and evaluated using receiver operator characteristic analysis to distinguish between PKD-C and PKD-D. Cortical-basal ganglia circuitry differed significantly between the 2 groups at a specific frequency. Furthermore, the PKD-D and PKD-C patients were observed to show different spontaneous brain activity in the right precuneus, right putamen, and right angular gyrus at the slow-5 frequency band (0.01-0.027 Hz). The frequency-specific abnormal local synchronization between the 2 types of PKD offers new insights into the pathophysiology of this disorder to some extent.

Globus Pallidus Interna in Tourette Syndrome: Decreased Local Activity and Disrupted Functional Connectivity.

Globus pallidus interna (GPi) is an effective deep brain stimulation site for the treatment of Tourette syndrome (TS), and plays a crucial role in the pathophysiology of TS. To investigate the functional network feature of GPi in TS patients, we retrospectively studied 24 boys with 'pure' TS and 32 age-/education-matched healthy boys by resting state functional magnetic resonance images. Amplitude of low-frequency fluctuation (ALFF) and functional connectivity were used to estimate the local activity in GPi and its functional coordinate with the whole brain regions, respectively. We found decreased ALFF in patients' bilateral GPi, which was also negatively correlated with clinical symptoms. Functional connectivity analysis indicated abnormal regions within motor and motor-control networks in patients (inferior part of sensorimotor area, cerebellum, prefrontal cortex, cingulate gyrus, caudate nucleus, and brain stem). Transcranial magnetic stimulation sites defined by previous studies ("hand knob" area, premotor area, and supplementary motor area) did not show significantly different functional connectivity with GPi between groups. In summary, this study characterized the disrupted functional network of GPi and provided potential regions-of-interest for further basic and clinical studies on TS.

Heat shock protein 70 is associated with replicase complex of Japanese encephalitis virus and positively regulates viral genome replication.

Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that causes the most prevalent viral encephalitis in Asia. The NS5 protein of JEV is a key component of the viral replicase complex, which plays a crucial role in viral pathogenesis. In this study, tandem affinity purification (TAP) followed by mass spectrometry analysis was performed to identify novel host proteins that interact with NS5. Heat shock protein 70 (Hsp70), eukaryotic elongation factor 1-alpha (eEF-1α) and ras-related nuclear protein (Ran) were demonstrated to interact with NS5. In addition to NS5, Hsp70 was also found to interact with NS3 which is another important member of the replicase complex. It was observed that the cytoplasmic Hsp70 partially colocalizes with the components of viral replicase complex including NS3, NS5 and viral dsRNA during JEV infection. Knockdown of Hsp70 resulted in a significantly reduced JEV genome replication. Further analysis reveals that Hsp70 enhances the stability of viral proteins in JEV replicase complex. These results suggest an important role for Hsp70 in regulating JEV replication, which provides a potential target for the development of anti-JEV therapies.