Identification of the Largest SCA36 Pedigree in Asia: with Multimodel Neuroimaging Evaluation for the First Time.

Spinocerebellar ataxias (SCAs) are a large group of hereditary neurodegenerative diseases characterized by ataxia and dysarthria. Due to high clinical and genetic heterogeneity, many SCA families are undiagnosed. Herein, using linkage analysis, WES, and RP-PCR, we identified the largest SCA36 pedigree in Asia. This pedigree showed some distinct clinical characteristics. Cognitive impairment and gaze palsy are common and severe in SCA36 patients, especially long-course patients. Although no patients complained of hearing loss, most of them presented with hearing impairment in objective auxiliary examination. Voxel-based morphometry (VBM) demonstrated a reduction of volumes in cerebellum, brainstem, and thalamus (corrected P < 0.05). Reduced volumes in cerebellum were also found in presymptomatic carriers. Resting-state functional MRI (R-fMRI) found reduced ReHo values in left cerebellar posterior lobule (corrected P < 0.05). Diffusion tensor imaging (DTI) demonstrated a reduction of FA values in cerebellum, midbrain, superior and inferior cerebellar peduncle (corrected P < 0.05). MRS found reduced NAA/Cr values in cerebellar vermis and hemisphere (corrected P < 0.05). Our findings could provide new insights into management of SCA36 patients. Detailed auxiliary examination are recommended to assess hearing or peripheral nerve impairment, and we should pay more attention to eye movement and cognitive changes in patients. Furthermore, for the first time, our multimodel neuroimaging evaluation generate a full perspective of brain function and structure in SCA36 patients.

Identification of genes associated with soluble sugar and organic acid accumulation in 'Huapi' kumquat (Fortunella crassifolia Swingle) via transcriptome analysis.

BACKGROUND: The levels and ratios of sugar and acid are important contributors to fruit taste. Kumquat is one of the most economically important citrus crops, but information on the soluble sugar and organic acid metabolism in kumquat is limited. Here, two kumquat varieties - 'Rongan' (RA) and its mutant 'Huapi' (HP) - were used to assess soluble sugar and organic acid accumulation and the related genes. RESULTS: Soluble sugars include sucrose, glucose and fructose, while malate, quinic acid and citrate are the dominant organic acids in the fruits of both kumquat varieties. HP accumulated more sugars but fewer organic acids than did RA. Transcriptome analysis revealed 63 and 40 differentially expressed genes involved in soluble sugar and organic acid accumulation, respectively. The genes associated with sugar synthesis and transport, including SUS, SPS, TST, STP and ERD6L, were up-regulated, whereas INVs, FRK and HXK genes related to sugar degradation were down-regulated in HP kumquat. For organic acids, the up-regulation of PEPC and NAD-MDH could accelerate malate accumulation. In contrast, high expression of NAD-IDH and GS resulted in citric acid degradation during HP fruit development. Additionally, the PK, PDH, PEPCK and FBPase genes responsible for the interconversion of soluble sugars and organic acids were also significantly altered in the early development stages in HP. CONCLUSION: The high sugar accumulation in HP fruit was associated with up-regulation of SUS, SPS, TST, STP and ERD6L genes. The PEPCK, PEPC, NAD-MDH, NADP-IDH, GS and FBPase genes played important roles in acid synthesis and degradation in HP kumquat. These findings provide further insight into understanding the mechanisms underlying metabolism of sugars and organic acids in citrus. © 2021 Society of Chemical Industry.

Genome-Wide Identification of Long Non-coding RNA in Trifoliate Orange (Poncirus trifoliata (L.) Raf) Leaves in Response to Boron Deficiency.

Long non-coding RNAs (lncRNAs) play important roles in plant growth and stress responses. As a dominant abiotic stress factor in soil, boron (B) deficiency stress has impacted the growth and development of citrus in the red soil region of southern China. In the present work, we performed a genome-wide identification and characterization of lncRNAs in response to B deficiency stress in the leaves of trifoliate orange (Poncirus trifoliata), an important rootstock of citrus. A total of 2101 unique lncRNAs and 24,534 mRNAs were predicted. Quantitative real-time polymerase chain reaction (qRT-PCR) experiments were performed for a total of 16 random mRNAs and lncRNAs to validate their existence and expression patterns. Expression profiling of the leaves of trifoliate orange under B deficiency stress identified 729 up-regulated and 721 down-regulated lncRNAs, and 8419 up-regulated and 8395 down-regulated mRNAs. Further analysis showed that a total of 84 differentially expressed lncRNAs (DELs) were up-regulated and 31 were down-regulated, where the number of up-regulated DELs was 2.71-fold that of down-regulated. A similar trend was also observed in differentially expressed mRNAs (DEMs, 4.21-fold). Functional annotation of these DEMs was performed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, and the results demonstrated an enrichment of the categories of the biosynthesis of secondary metabolites (including phenylpropanoid biosynthesis/lignin biosynthesis), plant hormone signal transduction and the calcium signaling pathway. LncRNA target gene enrichment identified several target genes that were involved in plant hormones, and the expression of lncRNAs and their target genes was significantly influenced. Therefore, our results suggest that lncRNAs can regulate the metabolism and signal transduction of plant hormones, which play an important role in the responses of citrus plants to B deficiency stress. Co-expression network analysis indicated that 468 significantly differentially expressed genes may be potential targets of 90 lncRNAs, and a total of 838 matched lncRNA-mRNA pairs were identified. In summary, our data provides a rich resource of candidate lncRNAs and mRNAs, as well as their related pathways, thereby improving our understanding of the role of lncRNAs in response to B deficiency stress, and in symptom formation caused by B deficiency in the leaves of trifoliate orange.

[Magnetic Resonance Imaging Features and Their Pathological Mechanisms of Hashimoto's Encephalopathy].

Objective To analyze the routine and functional magnetic resonance imaging(MRI) features and their potential pathological mechanisms of Hashimoto's encephalopathy(HE). Methods The clinical data and routine and functional MRI images of 30 HE patients who were treated in our center from January 2010 to April 2017 were retrospectively reviewed. Among them,15 patients were examined with contrast-enhanced MRI,16 with diffusion-weighed imaging(DWI),8 with magnetic resonance angiography,2 with magnetic resonance spectroscopy,and 1 with both arterial spin labeled perfusion imaging and diffusion tensor imaging. Seven patients had consecutive clinical and imaging data. The distribution,MRI signals,and functional MRI features of HE were analyzed. Results Among 30 HE patients,routine MRI showed negative results in 8 cases and abnormal findings in 22 cases. Among 22 abnormal cases,9 were characterized by small cerebral vascular disease and 13 had non-specific abnormalities;of these 13 cases,12 had lesions mainly located at the supratentorial white matter,11 had multiple lesions,and 2 had lesions complicated with cerebellum atrophy. The lesions were focal or confluent,punctate or small patchy,showing abnormal signal intensity with iso-or hypo-intensity on T1-weighed imaging,hyper-intensity on both T2-weighed imaging and fluid-attenuated inversion recovery. Most of the lesions had no enhancement(12/15). Among 7 cases with abnormalities on DWI,hyper-intensity on DWI and hypo-intensity on apparent diffusion coefficient were seen in 3 sudden acute cases and hyper-intensity on DWI and increased apparent diffusion coefficient value in 4 sub-acute or slow onset cases. Three cases showed localized intracranial artery stenosis. In 2 cases,magnetic resonance spectroscopy revealed significant lower N-acetylaspartate peak,higher choline peak,and visible lactate peak or lipid peak. Of 7 cases with follow-up data,3 cases had no change,4 cases had changes including softening lesions(2/4),remitted and relapsed lesions(1/4),and rapid progression of brain atrophy with negative finding on the initial MRI(1/4). Conclusion Routine MRI combined with functional imaging can show the features of HE from different perspectives. Routine MRI shows multifocal or confluent lesions in the white matter,mostly without enhancement,while functional imaging may reveal pathological characteristics of different phases of acute or chronic ischemia and demyelinating changes of HE. Combined with clinical data,MRI can differentiate HE from other diseases based on routine and functional MRI appearances.

[Influencing factors for duration of viral nucleic acid shedding in children with influenza A].

OBJECTIVE: To investigate the features and duration of viral nucleic acid shedding in children with influenza A. METHODS: The clinical data of 90 children with influenza A with positive influenza A virus nucleic acid in nasopharyngeal swab detected by PCR were collected, and these children were divided into simple influenza A group (n=10), influenza A-pneumonia group (n=61), influenza A-nervous system damage group (n=10), and influenza A-underlying disease group (n=9). A retrospective analysis was performed for clinical features, treatment process, duration of viral nucleic acid shedding, and prognosis. RESULTS: The most common symptoms in these children were fever (89/90, 99%), cough (89/90, 99%), running nose (69/90, 77%), shortness of breath (26/90, 29%), and myalgia (23/90, 26%). The mean duration of viral nucleic acid shedding in 90 children was 9.4±2.9 days. The simple influenza A group had a significantly shorter duration of viral nucleic acid shedding than the influenza A-pneumonia, influenza A-nervous system damage, and influenza A-underlying disease groups (p<0.05), while there were no significant differences between the influenza A-pneumonia, influenza A-nervous system damage, and influenza A-underlying disease groups (p>0.05). The children who received antiviral therapy within 48 hours after disease onset had significantly shorter duration of viral nucleic acid shedding and time to body temperature recovery than those who received antiviral therapy more than 48 hours after disease onset (p<0.05). Of all the children with body temperature recovery, 83% still tested positive for viral nucleic acid. CONCLUSIONS: Complications, underlying diseases, and timing of antiviral therapy are influencing factors for the duration of influenza A virus nucleic acid shedding, and whether body temperature returns to normal cannot be used to decide whether to continue antiviral therapy.

DCE-MRI assessment of the effect of Epstein-Barr virus-encoded latent membrane protein-1 targeted DNAzyme on tumor vasculature in patients with nasopharyngeal carcinomas.

BACKGROUND: EBV-encoded latent membrane protein 1 (EBV-LMP1) is an important oncogenic protein for nasopharyngeal carcinoma (NPC) and has been shown to engage a plethora of signaling pathways. Correspondingly, an LMP1-targeted DNAzyme was found to inhibit the growth of NPC cells both in vivo and in vitro by suppressing cell proliferation and inducing apoptosis. However, it remains unknown whether an LMP1-targeted DNAzyme would affect the vasculature of NPC. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) has been applied in the clinical trials of anti-angiogenic drugs for more than ten years, and Ktrans has been recommended as a primary endpoint. Therefore, the objective of the current study was to use DCE-MRI to longitudinally study the effect of an EBV-LMP1-targeted DNAzyme on the vasculature of patients with NPC. METHODS: Twenty-four patients were randomly divided into two groups: a combined treatment group (radiotherapy + LMP1-targeted DNAzyme) and a radiotherapy alone group (radiotherapy + normal saline). DCE-MRI scans were conducted 1 ~ 2 days before radiotherapy (Pre-RT), during radiotherapy (RT 50 Gy), upon completion of radiotherapy (RT 70 Gy), and three months after radiotherapy (3 months post-RT). Parameters of vascular permeability and intra- and extravascular volumes were subsequently obtained (e.g., Ktrans, kep, ve) using nordicICE software. RESULTS: Both Ktrans and kep values for NPC tumor tissues decreased for both groups after treatment. Moreover, a statistically significant difference in Ktrans values at the pre-therapy and post-therapy timepoints emerged earlier for the combined treatment group (RT 50 Gy, P =0.045) compared to the radiotherapy alone group (3 months post-RT, P = 0.032). For the kep values, the downward trend observed for both the combined treatment group and the radiotherapy alone group were similar. In contrast, ve values for all of the tumor tissues increased following therapy. CONCLUSIONS: The EBV-LMP1-targeted DNAzyme that was tested was found to accelerate the decline of Ktrans values for patients with NPC. Correspondingly, the LMP1-targeted DNAzyme treatments were found to affect the angiogenesis and microvascular permeability of NPC. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01449942. Registered 6 October 2011.

[BOLD-fMRI in sensory area and motor hand functional area with brain tumor in the central area].

OBJECTIVE: To explore the geomorphological performance, the characteristics of volume, and the largest signal intension of blood oxygenation level dependent functional magnetic resonance imaging (BOLD-fMRI) in brain tumors located in or closed to the central area. METHODS: We recruited 13 normal volunteers and 31(13 benign tumors and 18 malignant tumors) patients with brain tumor located in or closed to the central area, to examine both side hand motor and tactile function by BOLD-fMRI and obtained the activation map and its superposition image with T1 imaging, the volume, and the largest signal intension of the functional area by SPM software which manipulated the raw data in the off-line work station. The volume and the largest signal intension data of the functional area were statistically analyzed by SPSS 13.0. RESULTS: The volume and the largest signal intension of the activation map in the normal functional area had hemisphere advantage. There was difference in the activated signal pixel number and the largest signal intension of the functional area between the benign brain tumors, malignant brain tumors, and the normal volunteers (P < 0.05). The shape, anatomic location, the volume, and the largest signal intension of the functional area were changed in the patients with brain tumors. CONCLUSION: BOLD-fMRI is a valid method to assess the pre-surgical risk of patients with brain tumors, which can get the volume, the largest signal intension, the basic shape,and the anatomic location of the functional area.

[Oral ferric ammonium citrate solution in MRCP for diagnosing low-level obstructive jaundice].

OBJECTIVE: To evaluate the value of oral ferric ammonium citrate solution as a gastrointestinal contrast agent in diagnosing low-level obstructive jaundice. METHODS: Thirty-six patients who were suspected of low-level obstructive jaundice were performed with magnetic resonance cholangiopancreatography (MRCP) and conventional MRI before and after the administration of oral ferric ammonium citrate solution. The diagnostic accuracy for evaluating the site and the cause of obstruction was compared with other diagnostic modalities. RESULTS: The image qualities of single-slice and multi-slice MRCP were improved markedly. The accuracy of MRCP for evaluating the site of obstruction was 97.22%, which was superior to US (P<0.05). There were no significant differences among the MRCP, CT, and ERCP. CONCLUSION: Oral ferric ammonium citrate solution can significantly improve the image quality of MRCP. FAC-MRCP is a simple, safe, and noninvasive technique with excellent accuracy in the diagnosis of low-level obstructive jaundice.

[Expression of Ang2 and Tie2 and their relation with the angiogenesis of hepatocellular carcinoma in rats].

OBJECTIVE: To investigate the relationship between the expression of Ang2, Tie2 and the angiogenesis of hepatocellular carcinoma in rats. METHODS: Thirty-eight healthy male rats were randomly divided into 3 groups: 5 rats in the control group; 25 rats in the experimental group were equally divided into 5-day, 10-day, 15-day, 20-day, and 25-day groups; the other 8 rats were used as the supplement of the experimental group. An allogenic transplanted rat model of CBRH-7919 hepatocellular carcinoma in situ was established by immunosuppression. The expressions of Ang2 and Tie2 were detected by immunohistochemical staining in cancerous tissues of different developmental stages and liver tissues of the control group. At the same time, microvessel density was determined by anti-CD31 immunohistochemical staining. RESULTS: CBRH-7919 hepatocellular carcinoma models were successfully set up in 24 rats. The expression level of Ang2 and Tie2 in cancerous tissues was much higher than that of liver tissues of the control group (P <0.05). The overexpression of Ang2 was pristine and continuous in different developmental stages. The expressions of Ang2 and Tie2 positively correlated with microvessal density in hepatocellular carcinoma (P<0.05). CONCLUSION: The up-regulation of Ang2 and Tie2 may play important roles in the angiogenesis of hepatocellular carcinoma. Ang2 may participate in the start of angiogenesis of hepatocellular carcinoma.

Magnetic resonance T2 relaxation time at 7 Tesla associated with amyloid ß pathology and age in a double-transgenic mouse model of Alzheimer's disease.

The aim of this study was to better understand the effect of amyloid-ß plaques on magnetic resonance T2 relaxation time. We investigated these changes associated with age in an APP/PS1 mouse model of AD at 7 Tesla, combined with histology. Ten double-transgenic AD and ten wild type (WT) female mice (aged 12-20 months) were used in a cross-sectional study. Mean T2 values and standard deviations were calculated for each Regions of interest (ROIs) on T2 map. Immunohistochemistry for amyloid plaques and fluorescence staining with thioflavine S were performed of brain sections after imaging. The results showed that mean T2 values of the hippocampus, cortex, corpus callosum, and thalamus of older mice were significantly lower than of the younger. Compared to WT mice, the T2 values of the hippocampus, corpus callosum, and thalamus in younger AD mice were significantly greater, while the T2 values of the hippocampus and cortex in older AD mice were significantly less. Aß-40 immunohistochemistry and thioflavine S stainging were positive in the matched region both for younger and older AD mice, while neither Aß-40 nor thioflavine S were observed in WT mice. These findings suggest that regional T2 values of AD mice may decrease with age, and changes in T2 values in AD mice may be influenced by many factors besides amyloid-ß plaque accumulation. Furthermore, they support that the standard deviation of the mean T2 value should be considered as well as the mean.

Transcription profiles of boron-deficiency-responsive genes in citrus rootstock root by suppression subtractive hybridization and cDNA microarray.

Boron (B) deficiency has seriously negative effect on citrus production. Carrizo citrange (CC) has been reported as a B-deficiency tolerant rootstock. However, the molecular mechanism of its B-deficiency tolerance remained not well-explored. To understand the molecular basis of citrus rootstock to B-deficiency, suppression subtractive hybridization (SSH) and microarray approaches were combined to identify the potential important or novel genes responsive to B-deficiency. Firstly four SSH libraries were constructed for the root tissue of two citrus rootstocks CC and Trifoliate orange (TO) to compare B-deficiency treated and non-treated plants. Then 7680 clones from these SSH libraries were used to construct a cDNA array and microarray analysis was carried out to verify the expression changes of these clones upon B-deficiency treatment at various time points compared to the corresponding controls. A total of 139 unigenes that were differentially expressed upon B-deficiency stress either in CC or TO were identified from microarray analysis, some of these genes have not previously been reported to be associated with B-deficiency stress. In this work, several genes involved in cell wall metabolism and transmembrane transport were identified to be highly regulated under B-deficiency stress, and a total of 23 metabolic pathways were affected by B-deficiency, especially the lignin biosynthesis pathway, nitrogen metabolism, and glycolytic pathway. All these results indicated that CC was more tolerant than TO to B-deficiency stress. The B-deficiency responsive genes identified in this study could provide further information for understanding the mechanisms of B-deficiency tolerance in citrus.

Digital gene expression analysis of corky split vein caused by boron deficiency in 'Newhall' Navel Orange (Citrus sinensis Osbeck) for selecting differentially expressed genes related to vascular hypertrophy.

Corky split vein caused by boron (B) deficiency in 'Newhall' Navel Orange was studied in the present research. The boron-deficient citrus exhibited a symptom of corky split vein in mature leaves. Morphologic and anatomical surveys at four representative phases of corky split veins showed that the symptom was the result of vascular hypertrophy. Digital gene expression (DGE) analysis was performed based on the Illumina HiSeq™ 2000 platform, which was applied to analyze the gene expression profilings of corky split veins at four morphologic phases. Over 5.3 million clean reads per library were successfully mapped to the reference database and more than 22897 mapped genes per library were simultaneously obtained. Analysis of the differentially expressed genes (DEGs) revealed that the expressions of genes associated with cytokinin signal transduction, cell division, vascular development, lignin biosynthesis and photosynthesis in corky split veins were all affected. The expressions of WOL and ARR12 involved in the cytokinin signal transduction pathway were up-regulated at 1(st) phase of corky split vein development. Furthermore, the expressions of some cell cycle genes, CYCs and CDKB, and vascular development genes, WOX4 and VND7, were up-regulated at the following 2(nd) and 3(rd) phases. These findings indicated that the cytokinin signal transduction pathway may play a role in initiating symptom observed in our study.

[Value of proton magnetic resonance spectroscopy with two-dimensional chemical-shift imaging in evaluating brain gliomas].

OBJECTIVE: To study the value of proton magnetic resonance spectroscopy ((1)H-MRS) with two-dimensional (2D) chemical-shift imaging (CSI) in evaluating brain gliomas. METHODS: Thirty-six patients with gliomas received examinations with 2D-CSI. The VOI of MRS included the tumor, peritumoral edematous and nonedematous areas, and the contralateral normal tissue. The changes of the metabolites in different areas were determined using 2D-CSI (1)H-MRS with SE sequence and the metabolite ratios were calculated. RESULTS: Significant differences were found in the ratios of NAA/Cr, Cho/Cr and NAA/Cho between low-grade gliomas and contralateral normal brain tissue, and between high-grade gliomas and the contralateral normal tissue (P<0.01). The low-grade gliomas and high-grade gliomas differed significantly in the ratios of NAA/Cr, Cho/Cr and NAA/Cho (P<0.05). These ratios also showed significant differences between peritumoral edematous area and the glioma tissue, between the peritumoral edematous area and contralateral normal brain tissue (P<0.05), and between the peritumoral nonedematous area and the glioma tissue (P<0.05). Between the peritumoral nonedematous area and contralateral normal brain tissue, NAA/Cr and NAA/Cho ratios were significantly different (P<0.05) but the Cho/Cr ratio was similar (P>0.05). CONCLUSIONS: MRS with 2D-CSI can provide precise and effective evidences with high time resolution for glioma grading, assessment of peritumoral involvement and glioma therapies.