Regulation of LncRNA00067110 on Proliferation, Apoptosis and Melanins Production of B16-F10 Cells by Targeting Cabyr / 中国生物化学与分子生物学报

Long non-coding RNA (lncRNA) is a type of non-coding RNA with the more than 200 nucleotides. Several lncRNAs have been identified as the potential targets for cancer therapy. LncRNA00067110 is one of the differentially expressed genes in the transcriptome profiles of melanoma B16-F10 cells compared to normal mice melanocytes. To investigate whether lncRNA00067110 regulates the proliferation, apoptosis and melanogenesis of B16-F10 cells, the calcium-binding tyrosine phosphorylation regulated protein (Cabyr) target gene was predicted by LncTar and verified by dual luciferase activities. The regulating function of lncRNA00067110 was investigated by the analysis of transcriptome profiles and to detect the proliferation, apoptosis and melanin production of B16-F10 cells transfected by the overexpression plasmids of lncRNA00067110. The results showed that the relationship of lncRNA00067110 targeting Cabyr, the mRNA and protein levels of proliferation (MEK/ERK/MNK/CREB) and melanogenesis-related genes (TYR family and CREB) were significantly down-regulated, while the mRNA and protein levels of apoptosis-related genes (AKT and Bcl-2) were up-regulated in B16-F10 cells with lncRNA00067110 overexpression. The transcriptome profile of B16-F10 cells with lncRNA00067110 overexpression showed that 17 genes were differentially expressed, among which Cabyr was up-regulated. Furthermore, the effect of lncRNA00067110 on the phenotypes of cell proliferation and apoptosis were verified. The results suggested that lncRNA00067110 might be a novel target for the treatment of melanoma by targeting Cabyr, which regulate the expression of related genes to inhibit the proliferation and melanogenesis, as well as to induce the apoptosis of B16-F10 cells.

MicroRNA-100-5p Regulates Melanogenesis in Alpaca Melanocytes by Targeting FGF21 / 中国生物化学与分子生物学报

Although many microRNAs (miRNAs) are known to function as regulators of coat color and melanogenesis, the underlying molecular mechanisms of miR-100-5p governing melanogenesis were not completely known.The goal of this study was to determine the effect of miR-l()()-5p on melanogenesis in alpaca melanocytes.Fibroblast growth factor 21 (FGF21) is a predicted target gene of miR-100-5p and the luciferase reporter assay demonstrated that miR-100-5p regulates FGF21 by binding to its 3' untranslated region (3'UTR).In this study, alpaca melanocytes were transfected with miR-100-5p, inhibitor and negative control plasmid.Results showed that miR-100-5p overexpression significantly decreased mRNA and protein expression of FGF2\.Meanwhile, the ERK signal pathway was inhibited, with subsequent up-regulation of microphthalmia-associated transcription factor (MITF) , tyrosinase (TYR) and tyrosinase-related protein 2 (TYRP2), which increased melanin production.The results suggest that miR-100-5p may regulate melanogenesis by targeting FGF21 via extracellular regulated MAP kinase (ERK) signaling pathway.

DKK3 Regulates Melanogenesis in Melanocytes of Alpaca / 中国生物化学与分子生物学报

Dickkopf-3 (DKK3) , as a critical inhibitor of the Wnt/p-catenin signaling pathway, may he involved in melanogenesis.In the current study, we investigated the effects of DKK3 on melanogenesis in melanocytes of alpaca.Overexpression of DKK3 in alpaca melanocytes, the expression of Wntl, Lefl , Myc and the major target genes termed microphthalmia-associated transcription factor (M1TF) and its downstream genes, including tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1) and tyrosinase- related protein 2 (TYRP2) were significantly decreased at both mRNA and protein levels (P<0.05); total alkali melanin, pheomelanin and eumelanin were decreased by 80.30%, 72.17% and 64.60% (P <0.05), respectively.In contrast, in the melanocytes transfected with siRNA-DKK3 (a small interference RNA targeting DKK3) , the expression of Wntl, Lefl, Myc, MITF, TYR, TYRPl and TYRP2 were significantly increased at both mRNA and protein levels (P<0.05) ; total alkali melanin, pheomelanin and eumelanin were significantly increased by 1.65 folds, 1.25 folds and 1.21 folds (P< 0.05) , respectively.These results indicate that DKK3 regulates melanogenesis in alpaca melanocytes via the Wnt/p-catenin signaling pathway and down-regulates MITF.

TMEM106B Induces Melanogenesis by Regulating ERK/CREB Signaling Pathway / 中国生物化学与分子生物学报

The TMEM106B protein is a type-II transmembrane protein, which localizes in the endosome and lysosome of dendrites in primary neurons. TMEM106B is essential for maintaining and branching of dendrites, and thus regulates retrograde lysosomal trafficking of dendrites in primary neurons. Mammalian melanocytes are derived from neural cells, while melanosomes are originated from early endosome. However, the function of TMEM106B protein in melanocytes and its potential molecular mechanism in melanogenesis still remain unknown. Recently it was reported that transcription factor EB (TFEB) was the regulator of lysosome synthesis and TMEM106B protein overexpression promoted TFEB translocation into the nucleus. However, MITF (microphthalmia-associated transcription factor) and TFEB regulate each other in melanoma cells in vitro. Here in, plasmid containing gene for TMEM106B overexpression was transfected into melanocytes to investigate the regulation of TMEM106B on melanogenesis. The results showed that TMEM106B protein was localized in the cytoplasm of melanocytes. Compared with the negative control (NC), the mRNA levels of cyclic AMP-responsive element-binding protein (CREB) and MITF, especially CREB, were significantly increased in melanocytes with TMEM106B overexpression P< 0. 001). Western blot analysis showed that the expression of phosphorylated MAP kinase (p-ERK) was apparently increased (P<0.001) and resulted in the up-regulation of melanogenic regulatory proteins, including MITF, tyrosinase (TYR), tyrosinase-related protein-1 (TYRP1) and 2 (TYRP2). Masson-Fontana method showed that TMEM106B influenced the production of melanin in melanocytes. The spectrophotometry assay indicated that the amount of total melanin (ASM) (P<0. 001) and eumelanin (EM) (P<0. 05) were increased in alpaca melanocytes transfected with TMEM106B, while pheomelanin (PM) (P<0. 001) was decreased. These results demonstrated that TMEM106B played a vital role in melanogenesis in melanocytes by regulating ERK/CREB signaling pathway.

Characteristics of Electroencephalogram Power Spectrum Network on Uninjured Side of Post-stroke Aphasia / 中国康复理论与实践

Objective:To clarify the characteristics of electroencephalogram (EEG) power spectrum network of right hemisphere in patients with post-stroke aphasia (PSA) after left brain damaged. Methods:From December, 2018 to June, 2019, twelve PSA patients with left hemisphere injury were recruited and twelve healthy adults were matched as control group. China Rehabilitation Research Center aphasia examination (CRRCAE) was used to evaluated the linguistic function, and the EEG was collected. The functional connection characteristics of Alpha and Theta power spectrum were compared, and the correlation with language items was analyzed. Results:In PSA patients, the functional connection enhanced in Alpha and Theta frequency bands of central area, frontal, parietal and frontal-parietal joint areas, and decreased in Theta frequency bands of temporoparietal, parietal-occipital area, frontal, fronta-parietal and frontal central areas. The enhancement of alpha frequency functional connection from the right parietal occipital region to the central region was significantly correlated to the reduction of the expression ability (P < 0.05), while the weakening of theta frequency functional connection between the right parietal occipital region and the forehead and the center of the forehead was significantly correlated to the ability of speaking, reading, copying, dictation and calculation (r = -0.676~-0.717, P < 0.05). Conclusion:EEG power spectrum network can reflect the reorganization of right brain function network, and the change of right frontal parietal central network function connection may be closely related to PSA language injury and recovery.

Stimulation Mode and Model of Word-picture Language Task in Functional Magnetic Resonance Imaging Test for Post-stroke Aphasia (review) / 中国康复理论与实践

The cognitive neuroscience researches about post-stroke aphasia provide the interpretation of all aspects of linguistics. The word-picture research paradigm can be applied to assess different types of aphasia, in various ways of stimulation modes and models. It is more helpful combining functional magenetic resonance imaging to research the mechanism of brain damage and recovery objectively. The interactive application of language task and imaging has also become a new direction in the mechanism study of aphasia.

Functional Connectivities in Right Hemisphere for Post-stroke Aphasia: Based on Dual Stream Model / 中国康复理论与实践

Objective:To study the effect of right brain language network in post-stroke aphasia (PSA) patients with left hemisphere injury. Methods:From December, 2018 to June, 2019, twelve PSA patients with left hemisphere injury, and twelve matched healthy controls were recruited to accept rest-state functional magnetic resonance imaging (f-MRI) scan, and analyzed the characteristics of right brain function network with Dual Stream model. Results:There were two patients lost. Compared with the controls, for dorsolateral lingual pathway, the functional connections increased from superior marginal gyrus to middle frontal gyrus and inferior frontal gyrus of trigone in the patients, while those decreased from posterior central gyrus to inferior frontal gyrus of insula. For ventral lingual pathway, the functional connection increased from angular gyrus to orbital inferior frontal gyrus. For ventral and dorsolateral double-pathway, the functional connections increased from temporal lobe to lenticular pallidum and angular gyrus, from caudate nucleus to inferior frontal gyrus of insula, from lenticular putamen nucleus to middle frontal gyrus and trigonometry, while it decreased from superior marginal gyrus and temporal lobe to inferior frontal gyrus of insula. There was a negative correlation between the functional connection from inferior frontal gyrus to lenticular putamen and repeating (r = -0.720, P < 0.05), between the functional connection from inferior frontal gyrus to the caudate nucleus to speaking and repeating (r < -0.696, P < 0.05). In terms of network index, there were significant differences between the patients and the controls in both local and global indexes for language key brain area in right brain (|t| > 2.143, P < 0.05). Conclusion:The functional network has reorganized in right hemisphere of PSA patients. However, the increase of connection between language critical cortex and subcortical nuclei may play a role in improvement of language function.

Advance in Application of Electroencephalogram for Post-stroke Aphasia (review) / 中国康复理论与实践

Electroencephalogram (EEG) contains abundant physiological and pathological information. With the development of analysis, EEG can measure the neurodynamics at the sub-second level during impaired speech processing, providing a new perspective for revealing the occurrence, development and recovery mechanism of post-stroke aphasia. EEG plays an important role in the goal setting of rehabilitation for post-stroke aphasia, and serves as an evaluation of the efficacy for clinical rehabilitation.

Role of Working Memory in Lexical Access: An Event-related Potential Study Based on Word-picture Matchin Tasks / 中国康复理论与实践

Objective To observe the role of working memory in the process of language, and provide an objective evaluation for Chinese language rehabilitation.Methods From December, 2017 to June, 2018, 20 healthy subjects accepted word-picture matching tasks as the word and picture showed on the different (task 1) or same (task 2) screens, and their event-related potential of N400 were compared in matched and mismatched conditions.Results Forteen subjects were included finally. Task 1 induced N400 appeared earlier with higher amplitude and more activation in central parietal occipital region, followed larger N170 and P1 wave. There were significant differences in most cerebral regions between the two tasks in N400 amplitude difference of matched and mismatched conditions (t＞ 2.319, P ＜ 0.05).Conclusion Word-picture matching tasks may involve more language-related brain regions with the intervention of working memory, that might work better in process of vocabulary. The tasks simulating the scene of Chinese language rehabilitation can be used as an objective evaluation for clinical activities.