Genetic and clinical profile of 15 Chinese families with GDAP1-related Charcot-Marie-Tooth disease and identification of H256R as a frequent mutation.

BACKGROUND AND AIMS: Mutations in ganglioside-induced differentiation-associated protein 1 (GDAP1) cause axonal or demyelinating Charcot-Marie-Tooth disease (CMT) with autosomal dominant or recessive inheritance. In this study, we aim to report the genotypic and phenotypic features of GDAP1-related CMT in a Chinese cohort. METHODS: Clinical, neurophysiological, genetic data, and available muscle/brain imaging information of 28 CMT patients with GDAP1 variants were retrospectively collected. RESULTS: We identified 16 GDAP1 pathogenic variants, among which two novel variants c.980dup(p.L328FfsX25) and c.480+4T>G were first reported. Most patients (16/28) presented with AR or AD CMT2K phenotype. Clinical characteristics in our cohort demonstrated that the AR patients presented earlier onset, more severe phenotype compared with the AD patients. Considerable intra-familial phenotypic variability was observed among three AD families. Muscle atrophy and fatty infiltration in the lower extremity were detected by Muscle magnetic resonance imaging (MRI) scans in four patients. MRI showed two AR patients showed more severe muscle involvement of the posterior compartment than those of the anterolateral compartment in the calf. One patient carrying Q38*/H256R variants accompanied with mild periventricular leukoaraiosis. CONCLUSIONS: In this study, we conducted an analysis of clinical features of the GDAP1-related CMT patients, expanded the mutation spectrum in GDAP1 by reporting two novel variants, and presented the prevalent occurrence of the H256R mutation in China. The screening of GDAP1 should be particularly emphasized in Chinese patients with CMT2, given the incomplete penetrance and pathogenic inheritance patterns involving dominant and recessive modes.

Identification of calcium chelating peptides from peanut protein hydrolysate and absorption activity of peptide-calcium complex.

BACKGROUND: Peanut peptides have good chelating ability with metal ions. However, there are few studies on the chelation mechanism of peanut peptides with calcium and absorption properties of peptide-calcium complex. RESULTS: Peptides with high calcium chelating rate were isolated and purified from peanut protein hydrolysate (PPH), and the chelation rate of component F21 was higher (81.4 ± 0.8%). Six peptides were identified from component F21 by liquid chromatography-tandem mass spectrometry, and the frequency of acidic amino acids and arginine in the amino acid sequence was higher in all six peptides. Peanut peptide-calcium complex (PPH21-Ca) was prepared by selecting component F21 (PPH21). Ultraviolet analysis indicated that the chelate reaction occurred between peanut peptide and calcium ions. Fourier transform infrared analysis showed that the chelating sites were carboxyl and amino groups on the amino acid residues of peptides. Scanning electron microscopy revealed that the surface of peanut peptide had a smooth block structure, but the surface of the complex had a granular morphology. Caco-2 cell model tests revealed that the bioavailability of PPH21-Ca was 58.4 ± 0.5%, which was significantly higher than that of inorganic calcium at 37.0 ± 0.4%. CONCLUSION: Peanut peptides can chelate calcium ions by carboxyl and amino groups, and the peptide-calcium complex had higher bioavailability. This study provides a theoretical basis for the development of new calcium supplement products that are absorbed easily. © 2024 Society of Chemical Industry.

Effectiveness of internet-based nursing interventions for the treatment of patients with periodontitis.

BACKGROUND: Periodontitis, one of the most common oral diseases, is a chronic inflammatory condition occur in response to bacterial plaque biofilms. Plaque control and oral hygiene instructions are the most widely used and effective nonsurgical treatment for periodontitis, which is based on a partnership between patient and clinician and requires a life-long commitment. The objective of this study was to analyze the effectiveness of internet-based nursing interventions for the treatment of patients with periodontitis. The findings from this study may help to enhance the therapeutic outcomes for patients with periodontitis. METHODS: A total of 80 patients with periodontitis treated in Zhejiang Province Stomatology hospital from December 2021 to January 2023 were randomly selected and divided into control group and intervention group with 40 cases each. The control group was given routine oral health guidance and the intervention group received internet based nursing intervention. The periodontal pocket depth, percentage of periodontal pocket depth (PD) ≥ 4 mm, bleeding on probing (BOP)%, and self-efficacy scale for oral health care (SESS) were assessed and compared at four time points: initial visit, 6-8-weeks follow-up, 3-months follow-up, and 6-months follow-up. RESULTS: There was no significant difference between the two groups in terms of age, gender, initial visit PD, initial visit PD ≥ 4 mm (%), initial visit BOP (%), and initial visit SESS (P > 0.05). The intervention group showed a significantly decreased percentage of PD ≥ 4 mm at 6-8 weeks and 6-months follow-up compared to the control group (P < 0.05). The PD, BOP%, and SESS scores of the intervention group were significantly better than those of the control group at 6-months follow-up (P < 0.05). There was no statistically significant difference in patient satisfaction between the two groups. CONCLUSIONS: This study confirmed that the internet-based nursing intervention in conjunction with periodontal treatment was able to improve the periodontal pocket depth, gingival bleeding and the level of self-efficacy of patients, suggesting that it is necessary to carry out the extended oral hygiene instructions via internet-based platforms for the patients in clinical practice.

Downregulation of TOP2 modulates neurodegeneration caused by GGGGCC expanded repeats.

GGGGCC repeats in a non-coding region of the C9orf72 gene have been identified as a major genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. We previously showed that the GGGGCC expanded repeats alone were sufficient to cause neurodegeneration in Drosophila. Recent evidence indicates that GGGGCC expanded repeats can modify various gene transcriptomes. To determine the role of these genes in GGGGCC-mediated neurotoxicity, we screened an established Drosophila model expressing GGGGCC expanded repeats in this study. Our results showed that knockdown of the DNA topoisomerase II (Top2) gene can specifically modulate GGGGCC-associated neurodegeneration of the eye. Furthermore, chemical inhibition of Top2 or siRNA-induced Top2 downregulation could alleviate the GGGGCC-mediated neurotoxicity in Drosophila assessed by eye neurodegeneration and locomotion impairment. By contrast, upregulated Top2 levels were detected in Drosophila strains, and moreover, TOP2A level was also upregulated in Neuro-2a cells expressing GGGGCC expanded repeats, as well as in the brains of Sod1G93A model mice. This indicated that elevated levels of TOP2A may be involved in a pathway common to the pathophysiology of distinct ALS forms. Moreover, through RNA-sequencing, a total of 67 genes, involved in the pathways of intracellular signaling cascades, peripheral nervous system development, and others, were identified as potential targets of TOP2A to modulate GGGGCC-mediated neurodegeneration.

Diagnostic value of nerve conduction study in NOTCH2NLC-related neuronal intranuclear inclusion disease.

BACKGROUND AND AIMS: Neuronal intranuclear inclusion disease (NIID) is a rare progressive neurodegenerative disorder mainly caused by abnormally expanded GGC repeats within the NOTCH2NLC gene. Most patients with NIID show polyneuropathy. Here, we aim to investigate diagnostic electrophysiological markers of NIID. METHODS: In this retrospective dual-center study, we reviewed 96 patients with NOTCH2NLC-related NIID, 94 patients with genetically confirmed Charcot-Marie-Tooth (CMT) disease, and 62 control participants without history of peripheral neuropathy, who underwent nerve conduction studies between 2018 and 2022. RESULTS: Peripheral nerve symptoms were presented by 53.1% of patients with NIID, whereas 97.9% of them showed peripheral neuropathy according to electrophysiological examinations. Patients with NIID were characterized by slight demyelinating sensorimotor polyneuropathy; some patients also showed mild axonal lesions. Motor nerve conduction velocity (MCV) of the median nerve usually exceeded 35 m/s, and were found to be negatively correlated with the GGC repeat sizes. Regarding the electrophysiological differences between muscle weakness type (n = 27) and non-muscle weakness type (n = 69) of NIID, nerve conduction abnormalities were more severe in the muscle weakness type involving both demyelination and axonal impairment. Notably, specific DWI subcortical lace sign was presented in only 33.3% of muscle weakness type, thus it was difficult to differentiate them from CMT. Combining age of onset, distal motor latency, and compound muscle action potential of the median nerve showed the optimal diagnostic performance to distinguish NIID from major CMT (AUC = 0.989, sensitivity = 92.6%, specificity = 97.4%). INTERPRETATION: Peripheral polyneuropathy is common in NIID. Our study suggest that nerve conduction study is useful to discriminate NIID.

Impact of COVID-19 on patient follow-up during supportive periodontal therapy: a retrospective study based on phone call survey.

BACKGROUND: COVID-19 and the subsequent intermittent lockdown measures from 2020 to 2022 in China critically disrupted regular medical activities, including dental care. This study aimed to investigate the impact of COVID-19 on long-term follow-up at the Stomatology Hospital, Zhejiang University School of Medicine and to evaluate potential causes of loss to follow-up. METHODS: A total of 1062 patients with periodontitis who visited the hospital from January 2019 to June 2022 were included in this study, and patient information was collected retrospectively in the form of a telephone questionnaire. The questionnaire consisted of 19 questions in four areas: demographic characteristics, clinical periodontal parameters, oral hygiene habits, and follow-up-related open-ended questions (specific reasons for loss to follow-up, attitudes toward follow-up and suggestions for increasing participation in future follow-ups). Regression analysis of factors influencing the follow-up of patients with periodontitis were analyzed by regression analysis using R (v4.2.3) software. RESULTS: A total of 536 (50.47%) valid questionnaires were collected from 1062 patients. Personal factors (42.5%), instead of the COVID-19 epidemic (20.0%), were the main factors that impacted the loss to follow-up in long-term periodontal treatment, while work factors (19.8%), hospital factors (16.4%), and transportation or distance factors (14.7%) were all important factors. A family history of periodontitis [odds ratio (OR) = 0.567, 95% confidence interval (CI): 0.393, 0.817, p = 0.002], as well as frequent use of dental devices (OR = 0.540, 95% CI: 0.375, 0.777, p = 0.001), were significantly associated with a "negative" attitude toward follow-up visits. CONCLUSION: This survey suggests that the COVID-19 epidemic factor was an important cause contributed to the loss to follow-up during supportive periodontal therapy (SPT) among a variety of potential factors. Majority of patients had negative attitudes toward subsequent continued participation in supportive care.

[Determination of vitamin A, D and E in dishes by two-dimensional liquid chromatography].

OBJECTIVE: To establish a method for the simultaneous determination of vitamins A, D and E in dishes by two-dimensional liquid chromatography. METHODS: The samples were saponified and extracted with a mixture of ethyl acetate and n-hexane(3∶2, V/V)under the protection of antioxidants, and determined by two-dimensional liquid chromatography. The baseline separation of retinol, α-tocopherol, ß-tocopherol, γ-tocopherol, δ-tocopherol, and α-tocotrienols was achieved by using Alphasil pentafluorophenyl column(PFP, 150 mm×4.6 mm, 5 µm) as the one-dimensional column, and using water and methanol as mobile phases for gradient elution under the fluorescence detector. The separation detection of ergocalciferol and cholecalciferol from other impurities was achieved on the UV detector by Alphasil VC-C_(18) column(150 mm×4.6 mm, 3.5 µm) as the two-dimensional column. RESULTS: The baseline separation detection of retinol, ergocalciferol, cholecalciferol and five tocopherols was achieved. The compounds were linearly correlated within the set range, and the correlation coefficients were >0.999. The recovery rate of the method was between 85.4% and 106.4%. The detection limit of all-trans retinol was 0.7 µg/100 g, and the limit of quantitation was 2.4 µg/100 g. The limits of detection and quantification of tocopherols ranged from 1.1 to 2.5 µg/100 g and 3.6 to 8.3 µg/100 g. The detection limit of ergocalciferol and cholecalciferol was 0.3 µg/100 g, and the limit of quantification was 1.0 µg/100 g. In the end, finished dishes such as dry fried hairtail, braised mushroom, steamed egg with soy sauce, sweet and sour ribs were repeatedly measured by this method for six times, and the relative standard deviation was less than 10%. CONCLUSION: The method has the characteristics of simple operation, good repeatability, high accuracy and friendly to operators and ecological environment. It can realize the simultaneous typing detection of vitamins A, D and E in finished dishes.

ZmTE1 promotes plant height by regulating intercalary meristem formation and internode cell elongation in maize.

Maize height is determined by the number of nodes and the length of internodes. Node number is driven by intercalary meristem formation and internode length by intercalary cell elongation, respectively. However, mechanisms regulating establishment of nodes and internode growth are unclear. We screened EMS-induced maize mutants and identified a dwarf mutant zm66, linked to a single base change in TERMINAL EAR 1 (ZmTE1). Detailed phenotypic analysis revealed that zm66 (zmte1-2) has shorter internodes and increased node numbers, caused by decreased cell elongation and disordered intercalary meristem formation, respectively. Transcriptome analysis showed that auxin signalling genes are also dysregulated in zmte1-2, as are cell elongation and cell cycle-related genes. This argues that ZmTE1 regulates auxin signalling, cell division, and cell elongation. We found that the ZmWEE1 kinase phosphorylates ZmTE1, thus confining it to the nucleus and probably reducing cell division. In contrast, the ZmPP2Ac-2 phosphatase promotes dephosphorylation and cytoplasmic localization of ZmTE1, as well as cell division. Taken together, ZmTE1, a key regulator of plant height, is responsible for maintaining organized formation of internode meristems and rapid cell elongation. ZmWEE1 and ZmPP2Ac-2 might balance ZmTE1 activity, controlling cell division and elongation to maintain normal maize growth.

Phenotypic and transcriptomic responses of the shade-grown species Panax ginseng to variable light conditions.

BACKGROUND AND AIMS: Elucidating how plant species respond to variable light conditions is important to understand the ecological adaptation to heterogeneous natural habitats. Plant performance and its underlying gene regulatory network have been well documented in sun-grown plants. However, the phenotypic and molecular responses of shade-grown plants under variable light conditions have remained largely unclear. METHODS: We assessed the differences in phenotypic performance between Panax ginseng (shade-grown) and Arabidopsis thaliana (sun-grown) under sunlight, shade and deep-shade conditions. To further address the molecular bases underpinning the phenotypic responses, we compared time-course transcriptomic expression profiling and candidate gene structures between the two species. KEY RESULTS: Our results show that, compared with arabidopsis, ginseng plants not only possess a lower degree of phenotypic plasticity among the three light conditions, but also exhibit higher photosynthetic efficiency under shade and deep-shade conditions. Further comparisons of the gene expression and structure reveal that differential transcriptional regulation together with increased copy number of photosynthesis-related genes (e.g. electron transfer and carbon fixation) may improve the photosynthetic efficiency of ginseng plants under the two shade conditions. In contrast, the inactivation of phytochrome-interacting factors (i.e. absent and no upregulation of the PIF genes) are potentially associated with the observed low degree of phenotypic plasticity of ginseng plants under variable light conditions. CONCLUSIONS: Our study provides new insights into how shade-grown plants respond to variable light conditions. Candidate genes related to shade adaptation in ginseng provide valuable genetic resources for future molecular breeding of high-density planting crops.

Detection of changes in synaptic density in amyotrophic lateral sclerosis patients using 18 F-SynVesT-1 positron emission tomography.

BACKGROUND AND PURPOSE: Synaptic loss is well established as the major correlate of characteristic and consistent pathology in amyotrophic lateral sclerosis (ALS). We aimed to assess the possible discriminant diagnostic value of 18 F-SynVesT-1 positron emission tomography (PET) as a marker of ALS pathology and investigate whether specific synaptic density signatures are present in ALS with different subtypes. METHODS: Twenty-one patients with ALS and 25 healthy controls (HCs) were recruited. All participants underwent 18 F-SynVesT-1-PET. Synaptic density between ALS and HCs and between different ALS subgroups were compared. Correlation between synaptic density and clinical features in ALS was also analyzed. RESULTS: Low uptake distribution was found in the group comprising 21 ALS patients as compared with HCs in the right temporal lobe and the bilateral inferior frontal gyrus, anterior cingulate, and hippocampus-insula region. We also found low uptake in the bilateral superior temporal gyrus, hippocampus-insula, anterior cingulate, and left inferior frontal gyrus in ALS patients with cognitive impairment compared to HCs. Furthermore, compared to spinal onset ALS, bulbar onset ALS showed low uptake in the bilateral cingulate gyrus and high uptake in the bilateral superior temporal gyrus and left occipital lobe. No significant result was found in correlation analysis. CONCLUSIONS: This approach may provide a direct measure of synaptic density, and it therefore might represent a potentially useful biomarker for ALS diagnosis, as well as for estimating the cognitive decline and site of onset in ALS. 18 F-SynVesT-1-PET is presently not justified as a routine investigation to detect evidence of brain dysfunction leading to progression in ALS.

Phylogenomic and evolutionary dynamics of inverted repeats across Angelica plastomes.

BACKGROUND: Angelica L. (family Apiaceae) is an economically important genus comprising ca. One hundred ten species. Angelica species are found on all continents of the Northern Hemisphere, and East Asia hosts the highest number of species. Morphological characters such as fruit anatomy, leaf morphology and subterranean structures of Angelica species show extreme diversity. Consequently, the taxonomic classification of Angelica species is complex and remains controversial, as the classifications proposed by previous studies based on morphological data and molecular data are highly discordant. In addition, the phylogenetic relationships of major clades in the Angelica group, particularly in the Angelica s. s. clade, remain unclear. Chloroplast (cp) genome sequences have been widely used in phylogenetic studies and for evaluating genetic diversity. RESULTS: In this study, we sequenced and assembled 28 complete cp genomes from 22 species, two varieties and two cultivars of Angelica. Combined with 36 available cp genomes in GenBank from representative clades of the subfamily Apioideae, the characteristics and evolutionary patterns of Angelica cp genomes were studied, and the phylogenetic relationships of Angelica species were resolved. The Angelica cp genomes had the typical quadripartite structure including a pair of inverted repeats (IRs: 5836-34,706 bp) separated by a large single-copy region (LSC: 76,657-103,161 bp) and a small single-copy region (SSC: 17,433-21,794 bp). Extensive expansion and contraction of the IR region were observed among cp genomes of Angelica species, and the pattern of the diversification of cp genomes showed high consistency with the phylogenetic placement of Angelica species. Species of Angelica were grouped into two major clades, with most species grouped in the Angelica group and A. omeiensis and A. sinensis grouped in the Sinodielsia with Ligusticum tenuissimum. CONCLUSIONS: Our results further demonstrate the power of plastid phylogenomics in enhancing the phylogenetic reconstructions of complex genera and provide new insights into plastome evolution across Angelica L.

Prolonged maternal separation alters neurogenesis and synaptogenesis in postnatal dentate gyrus of mice.

OBJECTIVES: As a common model for adverse early experience and depression, maternal separation (MS) is always used to investigate the psychological disease. Despite extensive and strong evidence verified the depression-like state induced by MS, little is known about the specific mechanism of MS. Therefore, the present study aimed to investigate the neurobiology mechanism of the MS-induced depression-like state. METHODS: To verify the depression-like behaviors of offspring induced by MS, a series of behavioral tests were performed. Then, in vivo electroporation and three-dimensional reconstruction, combining with immunohistochemistry and BrdU labeling, were mainly used to explore the neurogenesis and synaptogenesis in postnatal dentate gyrus. RESULTS: Prolonged MS indeed induced the depression-like behaviors of offspring in adulthood. Surprisingly, learning and memory were enhanced by prolonged MS. Further investigation indicated that prolonged MS inhibited the proliferation of neural stem cells, impaired the survival, and altered the fate decision of newborn cells, whereas the total length and terminal tips of dendrite, and the spine density, especially thin spine, were significantly increased in prolonged MS mice. CONCLUSIONS: Our results elucidated that prolonged MS induced the depression-like state by impairing postnatal neurogenesis of dentate gyrus. Importantly, our results emphasized that prolonged MS increased the spine density, especially thin spine, by increasing the total length and number of terminal tips of dendrite, thereby enhancing learning and memory.

Genotype and phenotype distribution of 435 patients with Charcot-Marie-Tooth disease from central south China.

BACKGROUND AND PURPOSE: The purpose was to provide an overview of genotype and phenotype distribution in a cohort of patients with Charcot-Marie-Tooth disease (CMT) and related disorders from central south China. METHODS: In all, 435 patients were enrolled and detailed clinical data were collected. Multiplex ligation-dependent probe amplification for PMP22 duplication/deletion and CMT multi-gene panel sequencing were performed. Whole exome sequencing was further applied in the remaining patients who failed to achieve molecular diagnosis. RESULTS: Among the 435 patients, 216 had CMT1, 14 had hereditary neuropathy with pressure palsies (HNPP), 178 had CMT2, 24 had distal hereditary motor neuropathy (dHMN) and three had hereditary sensory and autonomic neuropathy (HSAN). The overall molecular diagnosis rate was 70%: 75.7% in CMT1, 100% in HNPP, 64.6% in CMT2, 41.7% in dHMN and 33.3% in HSAN. The most common four genotypes accounted for 68.9% of molecular diagnosed patients. Relatively frequent causes were missense changes in PMP22 (4.6%) and SH3TC2 (2.3%) in CMT1; and GDAP1 (5.1%), IGHMBP2 (4.5%) and MORC2 (3.9%) in CMT2. Twenty of 160 detected pathogenic variants and the associated phenotypes have not been previously reported. Broad phenotype spectra were observed in six genes, amongst which the pathogenic variants in BAG3 and SPTLC1 were detected in two sporadic patients presenting with the CMT2 phenotype. CONCLUSIONS: Our results provided a unique genotypic and phenotypic landscape of patients with CMT and related disorders from central south China, including a relatively high proportion of CMT2 and lower occurrence of PMP22 duplication. The broad phenotype spectra in certain genes have advanced our understanding of CMT.

Structure of the FERM domain of a neural scaffold protein FRMPD4 implicated in X-linked intellectual disability.

Scaffold proteins play crucial roles in orchestrating synaptic signaling and plasticity in the excitatory synapses by providing a structural link between glutamatergic receptors, signaling molecules, and neuronal cytoskeletons. FRMPD4 is a neural scaffold protein that binds to metabotropic glutamate receptors via its FERM domain. Here, we determine the crystal structure of the FERM domain of FRMPD4 at 2.49 Šresolution. The structure reveals that the canonical target binding groove of FRMPD4 FERM is occupied by a conserved fragment C-terminal to the FERM domain, suggesting that the FRMPD4-mGluR interaction may adopt a distinct binding mode. In addition, FRMPD4 FERM does not contain a typical phosphoinositide binding site at the F1/F3 cleft found in ERM family FERM domains, but it possesses a conserved basic residue cluster on the F2 lobe which could bind to lipid effectively. Finally, analysis of mutations that are associated with X-linked intellectual disability suggests that they may compromise the biological function of FRMPD4 by destabilizing the FERM structure.

Simultaneous separation and determination of four active ingredients in Picria fel-terrae Lour. and its preparations by micellar electrokinetic chromatography.

INTRODUCTION: Picfeltarraenins IA, IB and IV and acteoside are the four bioactive ingredients of Picria fel-terrae Lour. Their pharmacological effects include central inhibitory, cardiovascular, anti-inflammatory, anti-pyretic, analgesic, anti-bacterial, antioxidative and anti-tumor effects. OBJECTIVE: We aimed to develop an efficient micellar electrokinetic chromatography (MEKC) method modified with mixed organic solvents for the simultaneous separation and determination of the four components in Picriae Herba and its formulations. METHODS: Method optimization was carried out by investigating influences of significant factors on the separation, and this method was successfully applied for the determination of the four components in Picriae Herba and its formulations. RESULTS: The optimal running buffer was composed of 20 mM sodium tetraborate, 40 mM sodium cholate, 10% (v/v) methanol and 10% (v/v) isopropanol (pH 9.76). The separation voltage was 18 kV, the temperature was 25°C and the detection wavelength was 266 nm. Under the optimal separation conditions, the baseline separation of four components was achieved in less than 14 min. The correlation coefficients of the calibration curves were 0.9984-0.9995 for the analytes. The intraday and interday precision ranged from 1.5% to 2.5% and from 1.4% to 5.0%, respectively. Recoveries of analytes varied from 96.6% to 104.1%. CONCLUSION: The method was proved suitable for the determination of four components in Picriae Herba and its formulations. Good performance was obtained under optimal conditions, and the method provides an effective tool for the quality control of Picriae Herba and its formulations.

MALAT1 rs619586 polymorphism functions as a prognostic biomarker in the management of differentiated thyroid carcinoma.

This study aimed to explore the roles of miR-214 and MALAT1 rs619586 polymorphism in the control and survival of differentiated thyroid carcinoma (DTC) via Cox regression analyses. The levels of MALAT1, miR-214, and CTNNB1 in different experimental groups were compared to study the interaction among MALAT1, miR-214, and CTNNB1. MTT and colony assays were used to investigate the role of rs619586 polymorphism in cell growth. The G allele of rs619586 polymorphism obviously decreased the 5-year survival of patients with DTC. Additionally, compared with AA-genotyped patients, patients carrying the AG/GG genotypes of MALAT1 rs619586 polymorphism showed much higher levels of DTC grade and CTNNB1 expression, along with lower levels of MALAT1 and miR-214 expression. Furthermore, the transcription activity of MALAT1 was significantly lowered by the rs619586G allele or miR-214 mimic, while the miR-214 inhibitor upregulated the luciferase activity of MALAT1. Additionally, miR-214 inhibited CTNNB1 expression by targeting CTNNB1 3'-untranslated region. Finally, the G allele of MALAT1 rs619586 polymorphism apparently promoted cell proliferation. Our study indicated that miR-214 inhibited MALAT1 expression by directly binding to the G allele of MALAT1 rs619586 polymorphism, thus inhibiting CTNNB1 expression and promoting cell proliferation in the pathogenesis of DTC. Therefore, MALAT1 rs619586 polymorphism could be used to predict the prognosis of DTC.

Fragile X mental retardation protein modulates the stability of its m6A-marked messenger RNA targets.

N6-methyladenosine (m6A) is the most prevalent internal modification of mammalian messenger RNAs (mRNAs) and long non-coding RNAs. The biological functions of this reversible RNA modification can be interpreted by cytoplasmic and nuclear 'm6A reader' proteins to fine-tune gene expression, such as mRNA degradation and translation initiation. Here we profiled transcriptome-wide m6A sites in adult mouse cerebral cortex, underscoring that m6A is a widespread epitranscriptomic modification in brain. Interestingly, the mRNA targets of fragile X mental retardation protein (FMRP), a selective RNA-binding protein, are enriched for m6A marks. Loss of functional FMRP leads to Fragile X syndrome (FXS), the most common inherited form of intellectual disability. Transcriptome-wide gene expression profiling identified 2035 genes differentially expressed in the absence of FMRP in cortex, and 92.5% of 174 downregulated FMRP targets are marked by m6A. Biochemical analyses indicate that FMRP binds to the m6A sites of its mRNA targets and interacts with m6A reader YTHDF2 in an RNA-independent manner. FMRP maintains the stability of its mRNA targets while YTHDF2 promotes the degradation of these mRNAs. These data together suggest that FMRP regulates the stability of its m6A-marked mRNA targets through YTHDF2, which could potentially contribute to the molecular pathogenesis of FXS.

Larmor Precession: Observation and Utilization for Boosting the Signal Intensity of Radio Frequency Glow Discharge Mass Spectrometry.

A novel magnet array system was constructed to use Larmor precession for boosting the signal intensity of rf-GD-MS. The enhancement mechanism with four magnet array devices of a single-block magnet and 2 × 2, 3 × 2, and 3 × 4 magnet arrays was simulated and studied by COMSOL Multiphysics Software 5.4.0 (COMSOL) to determine if the electrons in the discharge plasma could perform Larmor precession along the direction perpendicular to the magnetic field. Induced by Larmor precession, inelastic collisions between the primary electrons and the sample produced numerous secondary electrons and further improved the ionization efficiency. Moreover, the fuzzy synthetic evaluation result predicted that the device with a 3× 2 magnet array would display the greatest enhancement effect among the four devices. On the basis of these theoretical studies, a magnet array system with four magnet array devices was fabricated and utilized for studies of two scintillation crystals BGO and PWO. The observations indicated that the signal intensities obtained for 209Bi and 208Pb with the magnet array system were 630-3600 times of that obtained without a magnet and were enhanced by a factor of 1.5-2.8 compared with a previously reported stacked magnetic device. Two NIST samples were used to validate the method, and the results suggested that relative errors were less than 10%, and the lowest detection limit for the 3 × 2 magnet array could reach 0.0032 µg·g-1. Furthermore, the magnet array enhancement system with Larmor precession offers an efficient and sensitive approach for direct analysis of nonconducting materials.

Simultaneous separation and determination of hirsutine and hirsuteine by cyclodextrin-modified micellar electrokinetic capillary chromatography.

INTRODUCTION: Hirsutine and hirsuteine are the main pharmacological activity ingredients of Uncaria rhynchophylla (UR), playing an important role in treating mental and cardiovascular diseases, such as Alzheimer's disease, hypertension, Parkinson's disease, potential anti-cancer activities and so on. OBJECTIVE: To develop a cyclodextrin-modified micellar electrokinetic capillary chromatography (CD-MEKC) method for the simultaneous separation and determination of hirsutine and hirsuteine from UR and its formulations. METHODOLOGY: The optimal method was developed by investigating influences of significant factors on the separation, and this method was successfully applied for the determination of hirsutine and hirsuteine in UR and its formulations. RESULTS: The optimal background electrolyte (BGE) consisted of 40 mM sodium dihydrogen phosphate (pH 7.0), 150 mM 2,6-dimethyl-ß-cyclodextrin (DM-ß-CD), 3 mM mono-(6-ethylenediamine-6-deoxy)-ß-cyclodextrin (ED-ß-CD), and 30 mM sodium cholate (SC). Under these conditions, hirsutine and hirsuteine were successfully separated within 13 min at the separation voltage of 15 kV, temperature of 25°C and the detection wavelength of 224 nm. For the analytes, linear calibration curves were performed within the range 5.0-160.0 µg/mL. The limit of detection (LOD, S/N = 3) and the limit of quantitation (LOQ, S/N = 10) were 0.41, 1.42 µg/mL for hirsutine and 0.60, 2.17 µg/mL for hirsuteine, respectively. The recoveries of three samples were from 97.9% to 102.3%. CONCLUSION: The method was successfully applied to the determination of hirsutine and hirsuteine in UR and its formulations. Meanwhile, it provides an effective reference of the quality control of UR and its formulations.

[Fat-soluble vitamins in different eggs and egg products in Hangzhou City].

OBJECTIVE: To analyze the contents of fat-soluble vitamins in different kinds of eggs and egg products in Hangzhou City. METHODS: The contents of fat-soluble vitamin A, vitamin E, vitamin K_1 and vitamin K_2(menaquinone-4, menaquinone-7 and menaquinone-9) in eggs and egg products were determined by the high-performance liquid chromatography method. The contents of vitamin D were determined by high performance liquid chromatography-tandem mass spectrometry. The determined contents were compared with the corresponding nutrient reference values. RESULTS: The contents of vitamin A, vitamin D, vitamin E and vitamin K in different eggs and egg products were 64-278 µg RAE/100 g edible, 0. 2-9. 6 µg/100 g edible, 0. 59-2. 31 mg α-TE/100 g edible and 9. 5-84. 8 µg/100 g edible, respectively, accounting for 4%-192% of the corresponding nutrient reference values. The contents of fat-soluble vitamin A, vitamin D, vitamin E and vitamin K in duck, goose and quail eggs were higher than those in chicken eggs and pigeon eggs. CONCLUSION: There are some differences in fat-soluble vitamin A, vitamin D, vitamin E and vitamin K in different eggs and egg products, but there is no significant difference between groups.