Nuclear m(6)A Reader YTHDC1 Regulates mRNA Splicing.

The regulatory role of N(6)-methyladenosine (m(6)A) and its nuclear binding protein YTHDC1 in pre-mRNA splicing remains an enigma. Here we show that YTHDC1 promotes exon inclusion in targeted mRNAs through recruiting pre-mRNA splicing factor SRSF3 (SRp20) while blocking SRSF10 (SRp38) mRNA binding. Transcriptome assay with PAR-CLIP-seq analysis revealed that YTHDC1-regulated exon-inclusion patterns were similar to those of SRSF3 but opposite of SRSF10. In vitro pull-down assay illustrated a competitive binding of SRSF3 and SRSF10 to YTHDC1. Moreover, YTHDC1 facilitates SRSF3 but represses SRSF10 in their nuclear speckle localization, RNA-binding affinity, and associated splicing events, dysregulation of which, as the result of YTHDC1 depletion, can be restored by reconstitution with wild-type, but not m(6)A-binding-defective, YTHDC1. Our findings provide the direct evidence that m(6)A reader YTHDC1 regulates mRNA splicing through recruiting and modulating pre-mRNA splicing factors for their access to the binding regions of targeted mRNAs.

The role of melanin-concentrating hormone-1 receptors in the voiding reflex in rats.

We have used the selective melanin-concentrating hormone-1 (MCH(1)) receptor antagonist SNAP 7941 [((+)-methyl (4S)-3-{[(3-{4-[3-(acetylamino)phenyl]-1-piperidinyl}propyl) amino]carbonyl}-4-(3,4-difluorophenyl)-6-(methoxymethyl)-2-oxo-1,2,3,4-tetrahydro-5-pyrimidinecarboxylate hydrochloride)] to investigate the role of the hypothalamic neuropeptide MCH in the control of voiding in rats. Intravenous administration of SNAP 7941 (3 and 10 mg/kg i.v.) produced dose-related inhibition of rhythmic, distension-induced voiding contractions in anesthetized rats. In conscious rats in which repeated voiding cycles were evoked by continuous slow transvesicular infusion of saline, intragastric SNAP 7941 [0.03-1 mg/kg intragastrically (i.g.)] produced sustained increases in infusion capacity (maximum = 220% basal), comparable with the effects of the 5-hydroxytryptamine(1A) antagonist WAY 100635 (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-2-pyridinyl-cyclohexanecarboxamide maleate salt), and the muscarinic antagonist, oxybutynin (4-diethylaminobut-2-ynyl 2-cyclohexyl-2-hydroxy-2-phenylacetate hydrochloride). SNAP 7941 produced similar results when administered at a low dose (0.01 nmol) into the lateral ventricle (intracerebroventricular). The opposite effect was produced when MCH (20 nmol) was delivered intracerebroventricularly, resulting in a 34% decrease in apparent bladder capacity with increased urinary frequency. The effect of MCH was blocked by the prior intragastric administration of SNAP 7941 (0.1 mg/kg), but oxybutynin (1 mg/kg) was ineffective. Finally, in conscious spontaneously hypertensive rats, SNAP 7941 (0.1 mg/kg i.g.) produced a 31% reduction in micturition frequency, accompanied by a 36% increase in bladder capacity, with no effect on total volume voided over 6 h. The data indicate that MCH acts via MCH(1) receptors within the CNS to modulate the voiding reflex in rats. The striking effects of the MCH(1) antagonist SNAP 7941 to increase bladder capacity and reduce voiding frequency indicate that MCH(1) antagonists may offer a potential novel approach for treating overactive bladder syndrome.

m(6)A RNA methylation is regulated by microRNAs and promotes reprogramming to pluripotency.

N(6)-methyladenosine (m(6)A) has been recently identified as a conserved epitranscriptomic modification of eukaryotic mRNAs, but its features, regulatory mechanisms, and functions in cell reprogramming are largely unknown. Here, we report m(6)A modification profiles in the mRNA transcriptomes of four cell types with different degrees of pluripotency. Comparative analysis reveals several features of m(6)A, especially gene- and cell-type-specific m(6)A mRNA modifications. We also show that microRNAs (miRNAs) regulate m(6)A modification via a sequence pairing mechanism. Manipulation of miRNA expression or sequences alters m(6)A modification levels through modulating the binding of METTL3 methyltransferase to mRNAs containing miRNA targeting sites. Increased m(6)A abundance promotes the reprogramming of mouse embryonic fibroblasts (MEFs) to pluripotent stem cells; conversely, reduced m(6)A levels impede reprogramming. Our results therefore uncover a role for miRNAs in regulating m(6)A formation of mRNAs and provide a foundation for future functional studies of m(6)A modification in cell reprogramming.

Mammalian WTAP is a regulatory subunit of the RNA N6-methyladenosine methyltransferase.

The methyltransferase like 3 (METTL3)-containing methyltransferase complex catalyzes the N6-methyladenosine (m6A) formation, a novel epitranscriptomic marker; however, the nature of this complex remains largely unknown. Here we report two new components of the human m6A methyltransferase complex, Wilms' tumor 1-associating protein (WTAP) and methyltransferase like 14 (METTL14). WTAP interacts with METTL3 and METTL14, and is required for their localization into nuclear speckles enriched with pre-mRNA processing factors and for catalytic activity of the m6A methyltransferase in vivo. The majority of RNAs bound by WTAP and METTL3 in vivo represent mRNAs containing the consensus m6A motif. In the absence of WTAP, the RNA-binding capability of METTL3 is strongly reduced, suggesting that WTAP may function to regulate recruitment of the m6A methyltransferase complex to mRNA targets. Furthermore, transcriptomic analyses in combination with photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) illustrate that WTAP and METTL3 regulate expression and alternative splicing of genes involved in transcription and RNA processing. Morpholino-mediated knockdown targeting WTAP and/or METTL3 in zebrafish embryos caused tissue differentiation defects and increased apoptosis. These findings provide strong evidence that WTAP may function as a regulatory subunit in the m6A methyltransferase complex and play a critical role in epitranscriptomic regulation of RNA metabolism.

FTO-dependent demethylation of N6-methyladenosine regulates mRNA splicing and is required for adipogenesis.

The role of Fat Mass and Obesity-associated protein (FTO) and its substrate N6-methyladenosine (m6A) in mRNA processing and adipogenesis remains largely unknown. We show that FTO expression and m6A levels are inversely correlated during adipogenesis. FTO depletion blocks differentiation and only catalytically active FTO restores adipogenesis. Transcriptome analyses in combination with m6A-seq revealed that gene expression and mRNA splicing of grouped genes are regulated by FTO. M6A is enriched in exonic regions flanking 5'- and 3'-splice sites, spatially overlapping with mRNA splicing regulatory serine/arginine-rich (SR) protein exonic splicing enhancer binding regions. Enhanced levels of m6A in response to FTO depletion promotes the RNA binding ability of SRSF2 protein, leading to increased inclusion of target exons. FTO controls exonic splicing of adipogenic regulatory factor RUNX1T1 by regulating m6A levels around splice sites and thereby modulates differentiation. These findings provide compelling evidence that FTO-dependent m6A demethylation functions as a novel regulatory mechanism of RNA processing and plays a critical role in the regulation of adipogenesis.

Training approaches of data literacy for teachers, undergraduates, scientific researchers and librarians / 中华医学图书情报杂志

After the connotation of data literacy was elaborated, the status quo of domestic and foreign data literacy education was described, the training approaches of data literacy for teachers, undergraduates, scientific researchers and librarians were analyzed, and their characteristics and differences were compared in order to provide reference and lessons for building the data literacy education system and implementing the data literacy education.

Investigation and analysis of the working status and development situation of psychotherapy in psychiatric hospitals / 上海交通大学学报（医学版）

Objective·To understand the current situation of psychotherapy applied and relevant personnel's working in psychiatric hospitals, and provide advice to facilitate and promote the integrated service team building of domestic mental health institutions and the industry development of psychological treatment. Methods·Ninety-six psychiatric hospitals in China were selected and self-compiled questionnaire was used during the investigation. The actual feedback was collected from 52 hospitals in 25 provinces. Results·① According to the classification criteria of ICD-10 mental disorder, patients mainly suffered from schizophrenia, schizotypal disorder and delusional disorder, followed by affective disorder, in the psychiatric hospitals. ② There were 50 (96.2％) hospitals with psychological outpatient clinics, and 37 (71.2％) hospitals were equipped with mental wards. ③ The main types of patients who came to the psychology department in the psychiatric hospitals were emotional disorders, neurosis, stress-related and physical form disorders. ④ The composition of psychotherapists was 6 physicians, 2 clinical psychologists, and 1 nurse. ⑤ The top five psychotherapy techniques used by the psychiatric hospitals were cognitive therapy, supportive psychotherapy, behavioral therapy, group therapy, and family therapy. ⑥ 44.2％ of the practitioners working in this filed thought that the income had been too low if they only engaged in psychological treatment. Conclusion·In the staffing of mental health agency, compared to previous studies, the number of the psychology specialists increases, but it still makes up a small percentage, which does not match the needs of outpatient and ward patients. Psychological practitioners who rely on psychological treatment alone are in a lower income, and the gap between the expected income and actual income is too large. Government, health administration departments and hospitals should provide more support to psychological services and promote the development of institutions and personnel.

T cell exhaustion induced by persistent mycobacterium bovis BCG purified protein derivative stimulation could be restored by a Chinese Herbal Bushen Jianpi Recipe in mice model / 中国免疫学杂志

Objective:To evaluate effects of Bushen Jianpi Recipe ( BSJP) on restoring of T cell exhaustion induced by persistent tuberculosis antigens in mice.Methods:The C57BL/6 mice were primed with BCG(5×106 CFU)and boosted with 20μg of Mycobacterium bovis BCG Purified Protein Derivative ( PPD ) once a week and lasted for 16 times to induce T cell exhaustion that happened in late stage of tuberculosis .BSJP was used to treat the mice with immune exhaustion for 3 weeks.Flow Cytometry was used to detect the numbers of CD 4+and CD8+T cells and PD-1 expression.ELISA was used to detect IL-2 and IFN-γsecretion.Bacteria load in lung tissue following BCG challeng was used to detect the protective capability of mice models .Results: Compared with the transient antigen stimulation group,the persistent antigen stimulation group had lower level of IFN-γand IL-2 secretions(P<0.01).The numbers of CD4+and CD8+T cells decreased and the level of inhibitory receptor PD-1 got higher(P<0.01).The protective efficacy against BCG challenging decreased ( P<0.01 ) .After treatment with BSJP , the production of cytokines IFN-γand IL-2 increased ( P<0.01 );the numbers of CD4+and CD8+T cells increased and PD-1 expression level on CD4+T cells decreased(P<0.01);the protective efficacy against BCG challenge increased (P<0.01).The results showed that BSJP can reduce the PD-1 expression,improve the IL-2 and IFN-γsecretion,and increase the protection against BCG challenge .Conclusion:Bushen Jianpi Recipe can restore T cell exhaustion induced by persistent tuberculosis antigen stimulation in mice , which gives a hope to overcome T cell exhaustion in tuberculosis with Chinese herbal in clinic.