Attention Deficit Hyperactivity Disorder (ADHD) is a clinically common neurodevelopmental disorder of the brain. In addition to genetic factors, an imbalance in gut flora may also play a role in the development of ADHD. Currently, it is critical to investigate the function of gut flora and related metabolites, which may form the fundamental basis of bidirectional cross-linking between the brain and the gut, in addition to focusing on the changed gut flora in ADHD. This study aimed to investigate the possible relationship between changes in gut flora and metabolites and ADHD by analyzing metagenome and untargeted metabolomics of fecal samples from ADHD patients. Specifically, we attempted to identify key metabolites and the metabolic pathways they are involved in, as well as analyze in detail the structure and composition of the gut flora of ADHD patients. In order to further investigate the relationship between gut flora and ADHD symptoms, some behavioral studies were conducted following the transplantation of gut flora from ADHD patients into rats. The results of the metagenome analysis revealed several distinct strains, including Bacteroides cellulosilyticus, which could be important for diagnosing ADHD. Additionally, the ADHD group showed modifications in several metabolic pathways and metabolites, including the nicotinamide and nicotinic acid metabolic pathways and the metabolite nicotinamide in this pathway. The behavioral results demonstrated that rats with ADHD gut flora transplants displayed increased locomotor activity and interest, indicating that the onset of behaviors such as ADHD could be facilitated by the flora associated with ADHD. This research verified the alterations in gut flora and metabolism observed in ADHD patients and provided a list of metabolites and flora that were significantly altered in ADHD. Simultaneously, our findings revealed that modifications to the microbiome could potentially trigger behavioral changes in animals, providing an experimental basis for comprehending the function and influence of gut flora on ADHD. These results might provide new perspectives for the development of novel treatment strategies.

Perfluorinated compounds (PFASs) are persistent environmental pollutants recognized for their bioaccumulation and potential neurotoxic effects, particularly in relation to attention deficit hyperactivity disorder (ADHD). This study investigated the effects of perinatal PFAS exposure on Sprague-Dawley (SD) rats, with a focus on behavioral outcomes and dopamine metabolism in the prefrontal cortex (PFC). Pregnant rats were administered PFASs at concentrations of 1 mg/L, 3 mg/L, and 10 mg/L through drinking water during gestation and lactation, with a control group included for comparison. The offspring were subjected to behavioral assessments, including open field, marble burying, and jump tests, to evaluate locomotor activity, attention, and impulsivity. Neurochemical analyses were conducted to measure the expression of tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (DDC), and dopamine receptor D1 (Drd1) in the PFC using Western blotting and quantitative PCR (qPCR). The results indicated that high-dose PFAS exposure (10 mg/L) resulted in hyperactivity, attention deficits, and impulsive behavior (p < 0.05), along with decreased TH and DDC expression and altered Drd1 levels, suggesting disrupted dopamine metabolism. These findings imply that perinatal PFAS exposure leads to behavioral abnormalities and disruptions in dopamine-related processes. ### Competing Interest Statement The authors have declared that no competing interests exist.

Purpose:Attention-Deficit/Hyperactivity Disorder (ADHD) is one of the most prevalent neurodevelopmental disorders in childhood, with a globally increasing incidence. This study aims to investigate alterations in prefrontal cortical lipid metabolism in ADHD model rats and following transcutaneous auricular vagus nerve stimulation (taVNS) intervention, and to elucidate the regulatory effects of the sphingosine kinase inhibitor SKI II on the SPHK-S1P signaling pathway. Methods:Lipidomic analysis was performed to profile the lipid spectrum in prefrontal cortex tissues from Wistar-Kyoto (WKY) control rats, spontaneously hypertensive rat (SHR) ADHD models, sham-operated rats, and taVNS intervention rats. Key sphingolipid metabolic enzymes were assessed by RT-qPCR and Western blot, while sphingosine-1-phosphate (S1P) levels were quantified via ELISA. ADHD model rats received intraperitoneal administration of SPHK inhibitor SKI II (15 mg/kg, 16 days). Behavioral tests evaluated hyperactivity, impulsivity, and anxiety-like phenotypes. Western blot analyzed expression of dopamine synthesis rate-limiting enzyme. Results:The ADHD model group exhibited significantly elevated ganglioside and lysophospholipid levels. taVNS intervention specifically reduced sphingomyelin content. SPHK1 and SPHK2 mRNA and protein expression were markedly upregulated in ADHD models, concomitant with increased S1P levels. taVNS selectively decreased Sphk2 mRNA expression (without altering protein levels). SKI II administration significantly ameliorated hyperactivity, impulsivity, and anxiety-like behaviors in ADHD models, concurrently restoring dopamine β-hydroxylase expression. Conclusion:The SPHK-S1P signaling axis is a core pathway driving sphingolipid metabolic dysregulation in the prefrontal cortex in ADHD. Targeted inhibition of this pathway synergistically modulates the expression levels of proteins associated with dopaminergic neurotransmission. The limited regulatory effect of taVNS on sphingolipid metabolism suggests its clinical benefits may stem from multi-pathway synergistic mechanisms.

Purpose. To identify gene signatures that are shared by autism spectrum disorder (ASD) and epilepsy (EP) and explore the potential molecular mechanism of the two diseases using WGCNA analysis. Additionally, to verify the effects of the shared molecular mechanism on ADHD, which is another neurological comorbidity. Methods. We screened the crosstalk genes between ASD and EP based on WGCNA and differential expression analysis from GEO and DisGeNET database and analyzed the function of the genes’ enrichment by GO and KEGG analyses. Then, with combination of multiple datasets and multiple bioinformatic analysis methods, the shared gene signatures were identified. Moreover, we explored whether the shared gene signature had influence on the other neurological disorder like ADHD by analyzing the difference of the relative genes’ expression based on bioinformatic analysis and molecular experiment. Results. By comprehensive bioinformatic analysis for multiple datasets, we found that abnormal immune response and abnormal lipid metabolic pathway played important roles in coincidence of ASD and EP. Base on the results of WGCNA, we got the hub genes in ASD and EP. In attention deficit and hyperactivity disorder (ADHD) animal model, we also found a significant difference of gene expression related to sulfatide metabolism, indicating that the abnormal sphingolipid metabolism was common in multiple neurological disorders. Conclusion. This study reveals shared gene signatures between ASD and EP and identifies abnormal sphingolipid metabolism as an important participant in the development of ASD, EP, and ADHD.

AIMS:Attention-deficit/hyperactivity disorder (ADHD) is a prevalent disorder of neurodevelopment in children. The diagnosis of ADHD mainly relies on the symptoms and some may be misdiagnosed due to age-based variation in behaviours. This study aimed to explore biomarkers that are greatly needed for the accurate diagnosis of ADHD.METHODS:Seven hundred and forty-two samples were retrospectively investigated in three independent cohorts, screening, training, and validation, for circulation microRNA measurement using microarray, Taqman polymerase chain reaction, and regression analysis.RESULTS:A panel of five miRNAs (miR-4516, miR-6090, miR-4763-3p, miR-4281, and miR-4466) were identified as ADHD independent risk factors that provided a high diagnostic accuracy and specificity of ADHD (AUC = 0.940 and 0.927 in the training and validation datasets, respectively). This panel of miRNAs differentiated ADHD well from control groups. After clinical improvement by treatment, the panel of miRNAs in patients and AUC changed significantly and were close to those in healthy controls. Importantly, the targets of the miRNAs identified were commonly enriched in receptor signalling pathways, ion channels, and synapse structures.CONCLUSION:Our study identified a useful panel of miRNAs that have considerable clinical value in evaluating ADHD and provide important evidence for aberrant epigenetic regulation in ADHD.

OBJECTIVE:In order to understand the role of long noncoding RNAs (lncRNAs) played in the mechanisms of glyphosate neurotoxicity in neuronal development.METHODS:Perinatal glyphosate exposure (PGE) mouse model was constructed, and a lncRNA microarray was used to study the lncRNA expression changes in the hippocampus tissue of perinatal glyphosate exposure mice. Then we used GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) databases to analyze the function of the differentially expressed mRNAs and lncRNAs.RESULTS:LncRNA microarray analysis revealed that 1759 lncRNAs and 759 mRNAs were differentially expressed in the perinatal glyphosate exposure (PGE) mice group (G group) compared with the normal control mice group (C group). The functions of the DEmRNAs are involved in the cellular response to hormone stimulus. The ceRNA analysis showed that some interaction networks existed, including (ENSMUST00000137546, ENSMUST00000160950)/(miR-34a-3p, miR-130a-3p)/(Il12b, Irf1). Further analysis of the target mRNAs of miRNAs indicated that the possible functions involved the neuroactive ligand-receptor interaction and calcium signaling pathway, which are involved in perinatal glyphosate exposure-induced neurotoxicity.CONCLUSION:The aberrant expression of lncRNAs is related to the perinatal glyphosate-exposed neurotoxicity. These lncRNAs affect the target gene expression level, might by regulating neuroactive ligand-receptor interactions. The (ENSMUST00000137546, ENSMUST00000160950)/ (miRNA-34a-5p, miR-130a-3p) / mRNAs (e.g., Il12b, Irf1) interaction network may functions in perinatal glyphosate exposure-induced neurotoxicity.

目的 采用生物信息学方法分析miR-30d-5p在注意缺陷多动障碍(ADHD)发病机制中的作用.方法 通过TargetScan7.2、miRDB和miRTarBase数据库预测miR-30d-5p的靶基因,利用Cytoscape 3.7.1软件中的ClueGO插件对靶基因进行GO和KEGG富集分析,同时通过Cytoscape 3.7.1软件中的STRING插件构建靶蛋白互作(PPI)网络并筛选出核心基因,通过与ADHD gene数据库中已验证的ADHD相关基因对比,分析miR-30d-5p在ADHD中的作用和意义.结果 GO富集显示miR-30d-5p靶基因定位到核质、细胞内细胞器、轴突、细胞膜等细胞结构,并在神经系统中广泛参与到神经元发育、分化、神经元发生的调节、神经元投射发育、顺式调控区结合及顺式调控区序列特异性DNA结合等生物过程.KEGG富集显示miR-30d-5p靶基因显著富集在FoxO信号通路、轴突导向、海马长时程增强和神经营养信号通路.NOTCH1、KRAS、SIRT1、MAPK8、SOCS3、NEDD4、SKP2、BDNF、NEDD4L、SOCS1为miR-30d-5p预测靶基因的PPI网络中的核心基因.与ADHD gene数据库对比结果显示miR-30d-5p预测靶基因有33个基因与ADHD相关,且33个基因中含有预测核心基因BDNF.结论 miR-30d-5p通过靶向调控BDNF基因参与ADHD发生发展的过程,miR-30d-5p有潜力成为ADHD诊疗相关的靶点.

OBJECTIVE:To study the association of microRNA expression before and after drug therapy with clinical symptoms in children with attention deficit hyperactivity disorder (ADHD). METHODS:A total of 80 previously untreated children with ADHD who were diagnosed from May 2017 to October 2018 were enrolled. The children who were willing to receive drug therapy were randomly divided into concerta-treated group with 31 children and strattera-treated group with 33 children. The children who were unwilling to receive treatment were enrolled as the untreated group with 16 children. A total of 60 children who underwent physical examination during the same period of time were enrolled as the healthy control group. SNAP-V score was determined at initial diagnosis and 3 and 6 months of follow-up. Serum samples were collected from the children with ADHD and the healthy control group. Quantitative real-time PCR was used to measure the relative expression of miR-4566-3p and miR-7641. RESULTS:The repeated measures analysis of variance showed that the SNAP-V score of attention deficit symptoms were different among the two treatment groups and the untreated group at the first visit and 3 months and 6 months after treatment (P<0.05). There were significant differences in the relative expression of the two miRNAs among the two treatment groups and the healthy control group at the first visit and 3 months and 6 months after treatment (P<0.05). The SNAP-V score of attention deficit symptoms and the relative expression of the two miRNAs were different in different time points in the subjects (P<0.05). There were interactions between grouping and time factors in the SNAP-V score of attention deficit symptoms and the relative expression of the two miRNAs (P<0.05). The SNAP-V score of hyperactive impulsive symptoms was different in different time points in the two treatment groups and the untreated group (P<0.05), but the significant difference in the score was not observed between two treatment groups and the untreated group (P>0.05), and there was no interaction between the time factor and the grouping factor (P>0.05). The SNAP-V score of attention deficit symptoms was negatively correlated with the relative expression of miRNA-4655-3p and miRNA-7641 (r=-0.314, -0.495 respectively; P<0.05) in ADHD children after drug treatment. CONCLUSIONS:Drug therapy can significantly improve the clinical symptoms of children with ADHD. The expression of miR-4655-3p and miR-7641 in serum can be used as biomarkers for the diagnosis and outcome evaluation of ADHD.

目的 采用基因芯片技术筛选注意缺陷多动障碍(attention-deficit hyperactivity disorder,ADHD)患儿外周血清中的差异非编码小RNA(microRNA,miRNA).方法 收集ADHD患儿血清样本20例,正常儿童60例,根据性别相同、年龄、身高和体重相仿的标准配对形成1:3的病例组.采用含有2549种人类miRNA探针的安捷伦公司的人miRNA芯片V21.0(agilent human miRNA microarray V21.0)获取ADHD患儿与正常儿童的miRNA谱,将检测到的miRNA信号值进行配对T检验分析,筛选出表达差异有统计学意义的miRNA,然后通过实时荧光定量PCR技术验证芯片结果.结果 芯片筛选结果显示,和正常儿童比较,ADHD患儿血清中miR-30d-5p、miR-19b-3p、miR-6085、miR-19b-3p、miR-6749-5p、miR-6826-5p、miR-3960等61个miRNA比较,差异有统计学意义(P＜0.05),其中fold change＞2倍的有miR-30d-5p(fold change=0.48,P=0.002)、miR-4655-3p (fold change =0.43,P=0.002)和miR-7641(fold change=0.47,P=0.02),这些差异表达的miRNA在20例ADHD患儿外周血清中均降低,选择改变量超过两倍的miR-30d-5p,miR-4655-3p,miR-7641进行实时荧光定量PCR验证,结果与芯片一致.结论 与正常儿童比较,ADHD患儿外周血清中,miR-30d-5p,miR-4655-3p和miR-7641表达量降低,并且可能成为诊断ADHD的潜在指标.

Cerebral ischemia causes severe brain injury and results in selective neuronal death through programmed cell death mechanisms, including apoptosis and autophagy. Minimizing neuronal injury has been considered a hot topic among clinicians. The present study elucidated the effect of thymosin β4 (Tβ4) on neuronal death induced by cerebral ischemia/reperfusion in PC12 cells that were subjected to oxygen‑glucose deprivation and reoxygenation (OGD/R). The survival, apoptotic and autophagy rates of PC12 cells were investigated. Tβ4 pre‑conditioning prior to OGD/R was performed to evaluate PC12‑cell viability and the protective mechanisms of Tβ4. Tβ4 significantly increased cell survival after OGD/R. Tβ4 inhibited the release of lactate dehydrogenase, downregulated malondialdehyde and upregulated the activities of glutathione peroxidase and superoxide dismutase. In addition, Tβ4 attenuated OGD/R‑associated decreases in the expression of P62 and the anti‑apoptotic protein B‑cell lymphoma‑2, as well as the upregulation of autophagy mediators, including autophagy‑related protein‑5 and the ratio of microtubule‑associated protein 1 light chain 3 (LC3) II vs. LC3 I. These results suggested that Tβ4 effectively inhibits cell apoptosis and autophagy induced by OGD/R. To the best of our knowledge, the present study was the first to report on the antioxidant, anti‑apoptotic and anti‑autophagic effects of Tβ4 in neuronal‑like PC12 cells. These results suggested that Tβ4 may be explored as a potential treatment for cerebral ischemia.

Glyphosate is the active ingredient in numerous herbicide formulations. The roles of glyphosate in embryo-toxicity and neurotoxicity have been reported in human and animal models. Recently, several studies have reported evidence linking neurodevelopmental disorders (NDDs) with gestational glyphosate exposure. However, the role of glyphosate in neuronal development is still not fully understood. Our previous study found that perinatal glyphosate exposure resulted in differential microRNA expression in the prefrontal cortex of mouse offspring. However, the mechanism of glyphosate-induced neurotoxicity in the developing brain is still not fully understood. Considering the pivotal role of Circular RNAs (circRNAs) in the regulation of gene expression, a circRNA microarray method was used in this study to investigate circRNA expression changes in the hippocampus of mice with perinatal glyphosate exposure. The circRNA microarrays revealed that 663 circRNAs were significantly altered in the perinatal glyphosate exposure group compared with the control group. Among them, 330 were significantly upregulated, and the other 333 were downregulated. Furthermore, the relative expression levels of mmu-circRNA-014015, mmu-circRNA-28128 and mmu-circRNA-29837 were verified using quantitative real-time polymerase chain reaction (qRT-PCR). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses demonstrated that stress-associated steroid metabolism pathways, such as aldosterone synthesis and secretion pathways, may be involved in the neurotoxicity of glyphosate. These results showed that circRNAs are aberrantly expressed in the hippocampus of mice with perinatal glyphosate exposure and play potential roles in glyphosate-induced neurotoxicity.

目的 研究自身免疫性肝炎(AIH)小鼠模型中辅助性T淋巴细胞(Th)22细胞水平及其功能因子IL-22的表达情况,探索其在AIH发病中的意义.方法 选取30只4周龄雄性C57BL/6小鼠随机分为对照组(n=6)、AIH组(n=14),剩余10只用于提取肝脏特异性抗原S100.在实验第1天和第7天通过腹腔注射法将新鲜提取的S100与等体积弗氏完全佐剂混合液注射到小鼠腹腔内,第28天成功建立AIH小鼠模型(造模期间AIH组有7只小鼠因注射不当、出现腹水、感染等原因死亡)后,将小鼠经腹腔注射5％水合氯醛(0.1 ml/10 g)麻醉后摘取眼球取血,收集小鼠脾脏采用流式细胞术检测Th22细胞数,RT-PCR检测AHR mRNA水平;收集肝组织用于HE染色观察肝脏病理变化,RT-PCR和免疫印迹法检测肝组织IL-22、IL-22R表达情况;ELISA法检测血清中IL-22细胞因子水平.2组间比较采用t检验.结果 对照组小鼠肝组织排列整齐,肝小叶结构清晰,AIH组小鼠汇管区有大量炎症细胞浸润,肝组织出现点状甚至碎片状坏死.AIH组Th22细胞数较对照组明显升高[(0.083 ±0.052)％ vs(1.555±0.812)％,t=-4.405,P＜0.05].AIH组小鼠脾脏中Th22转录因子AHR mRNA的相对表达量较对照组有明显升高(0.485±0.096 vs l.268±0.366,t=-5.452,P＜0.05).AIH组肝组织中IL-22 mRNA和IL-22R mRNA的相对表达量均较对照组明显升高(IL-22 mRNA:1.146±0.227 vs 3.813±0.478,t=-12.458,P＜0.001;IL-22R mRNA:0.276 ±0.037 vs 1.734 ±0.248,t=-15.333,P＜0.001);AIH组肝组织的IL-22及IL-22R的蛋白相对表达量均明显高于对照组(1.040±0.261 vs 0.410±0.077,t=-6.324,P＜0.05;1.432±0.304 vs 0.830±0.146,t=-6.316,P＜0.05).AIH组血清IL-22水平较对照组升高,且差异有统计学意义(t=-15.799,P＜0.05).结论 AIH小鼠Th22细胞及IL-22因子的表达水平均明显升高,其可能在AIH的发病中起重要作用.

Glyphosate is the active ingredient in numerous herbicide formulations. The role of glyphosate in neurotoxicity has been reported in human and animal models. However, the detailed mechanism of the role of glyphosate in neuronal development remains unknown. Recently, several studies have reported evidence linking neurodevelopmental disorders (NDDs) with gestational glyphosate exposure. The current group previously identified microRNAs (miRNAs) that are associated with the etiology of NDDs, but their expression levels in the developing brain following glyphosate exposure have not been characterized. In the present study, miRNA expression patterns were evaluated in the prefrontal cortex (PFC) of 28 postnatal day mouse offspring following glyphosate exposure during pregnancy and lactation. An miRNA microarray detected 55 upregulated and 19 downregulated miRNAs in the PFC of mouse offspring, and 20 selected deregulated miRNAs were further evaluated by quantitative polymerase chain reaction (PCR). A total of 11 targets of these selected deregulated miRNAs were analyzed using bioinformatics. Gene Ontology (GO) terms associated with the relevant miRNAs included neurogenesis (GO:0050769), neuron differentiation (GO:0030182) and brain development (GO:0007420). The genes Cdkn1a, Numbl, Notch1, Fosl1 and Lef1 are involved in the Wnt and Notch signaling pathways, which are closely associated with neural development. PCR arrays for the mouse Wnt and Notch signaling pathways were used to validate the effects of glyphosate on the expression pattern of genes involved in the Wnt and Notch pathways. Nr4a2 and Wnt7b were downregulated, while Dkk1, Dixdc1, Runx1, Shh, Lef-1 and Axin2 were upregulated in the PFC of mice offspring following glyphosate exposure during pregnancy and lactation. These results indicated abnormalities of the Wnt/β-catenin and Notch pathways. These findings may be of particular interest for understanding the mechanism of glyphosate-induced neurotoxicity, as well as helping to clarify the association between glyphosate and NDDs.

Sialic acid modification is a kind of post-translational modification. To investigate the regulation effect of sialic acid on neural differentiation, we used CycloManN propanyl perac (CycloManN pro), a metabolic precursor of sialic acid, to treat PC12 cells. We noted that CycloManN pro indeed robustly promoted global sialylation detected by MAL II lectin blot in PC12 cells. Simultaneously, we interestingly found that the neurite outgrowth of PC12 cells was significantly promoted by the CycloManN pro treatment. The profile analysis of sialylated proteins showed that a protein band at 55KD was greatly enhanced especially in PC12L cells after CycloManN pro treatment. After enrichment with lectin MAL II, the proteins in this band were analyzed by mass spectrometry. The results showed that 23 proteins were in the band, but the score of vimentin was the highest among them. To investigate further the role of vimentin in the process of neurite differentiation, vimentin construct was transfected into PC12 cells. We interestingly observed that ectopic expression of vimentin significantly enhanced the neurite outgrowth induced by CycloManN pro. However, after three potential glycosylation sites (Ser-7, Thr-33, Ser-34:) of vimentin were mutated to alanine, overexpression of the mutated vimentin completely lost the enhancement activity for the neural differentiation even in the presence of CycloManN pro. Taken together, our study demonstrated that vimentin was important in the induction of neural differentiation by CycloManN pro.

Attention deficit hyperactivity disorder (ADHD) is a child developmental and behavioral disorder which seriously hinders their education and development. To investigate the key regulators in the prefrontal cortex (PFC), the major affected areas of ADHD, microRNA (miR)-138,138*, 34c*, 296, and 494, were noted for their significant downregulation in ADHD model rats spontaneously hypertensive rats (SHRs) compared to Wistar Kyoto (WKY) rat control. Based on promoter sequence analysis and activity assay, glucocorticoid receptor (Nr3c1) was identified for the inhibition of the promoter activity of miR-138-1, 34c*, 296, and 494 genes and their transcription. In the PFC of ADHD model rats SHR, Nr3c1 expression was abnormally elevated and reversely correlated with the levels of miR-138-1, 34c, 296, and 494 expression. Luciferase report assays indicated that all miR-138, 138*, 34c*, 296, and 494 targeted the 3' untranslated region of transcription factor Bhlhb2 (Bhlhe40) messenger RNA (mRNA) in common and ectopic expression of miR-138,138*, 34c*, 296, and 494 further suppressed the expression of Bhlhb2 gene. Consistently, Bhlhb2 expression was significantly higher in PFC of ADHD model SHR than control. Overexpressed Bhlhb2 in vitro significantly suppressed PC12 cell differentiation, and silence of Bhlhb2 enhanced the growth of neurite axon and dendrite. To observe the roles of Bhlhb2 further in vivo, Bhlhb2 was silenced in the PFC of nine SHR rats. Interestingly, knockdown of Bhlhb2 significantly improved the hyperactivity behaviors in SHRs compared to control. These findings show that Nr3c1-Bhlhb2 axis dysregulation was involved in the development of attention deficit and hyperactivity.

目的 探究波形蛋白分子在PC12细胞神经分化中的作用.方法 用免疫印迹方法检测PC12H和PC12L细胞中波形蛋白的表达情况.用免疫印迹方法检测PC12H细胞和PC12L细胞在NGF处理前后波形蛋白的表达情况.构建波形蛋白干扰质粒,用免疫印迹方法检测该质粒的干扰效果.分别将对照质粒、波形蛋白干扰质粒瞬时转染PC12H细胞,培养0、1、2、3天观察与记录细胞的分化情况,用Image-pro plus软件定量分析这两组细胞的平均神经突长度和含分化神经突细胞的百分比例.结果 PC12H细胞中波形蛋白的表达明显高于PC12L细胞(P=0.000).NGF处理后PC12H和PC12L细胞中波形蛋白的表达量均增加,且在PC12L细胞中更明显(P=0.000).波形蛋白干扰质粒能有效地抑制PC12H细胞中波形蛋白的表达(P=0.000).瞬时转染波形蛋白干扰质粒能有效地抑制PC12H细胞的神经分化,定量分析结果显示干扰组细胞的平均神经突长度和含分化神经突细胞的百分比均明显小于对照组,差异均有统计学意义(P =0.000).结论 波形蛋白分子可以促进PC12细胞的神经分化.

Deregulation of long noncoding RNAs (lncRNAs) is becoming recognized as a major feature of many neurological disorders. In the current study, we aimed to measure the expression of seven lncRNAs and lncRNA-targeted mRNAs in the peripheral lymphocytes of attention-deficit/hyperactivity disorder (ADHD), autism spectrum disorder (ASD), and intellectual disability (ID) patients via quantitative real-time reverse transcriptase PCR (qRT-PCR). We found that in ADHD, the expression of mRNAs of the BDNF, SHANK2, HOXB5, and HOXA6 genes was down-regulated significantly, but there was no difference in the expression of selected lncRNAs. As for ASD, the expression of the mRNAs of the HOXA6 and HOXA13 genes was significantly down-regulated, accompanied by a significant reduction of lncRNA that overlaps with the gene locus of HOXA13. A gender-dependent difference in expression of lncRNAs and targeted mRNAs was indicated in ID. In male ID, there was a significantly down-regulated expression of mRNAs of HOXB5 and HOXA13, accompanied by differentially decreased expression of lncRNA-targeted mRNAs of SYT15, PKNOX2, SHANK2, HOXB5, HOXA6, and HOXA13. In female ID, the mRNAs of HOXB5 and HOXA6 were significantly down-regulated, with the significantly down-regulated expression of lncRNA-targeted mRNAs of BDNF, PKNOX2, HOXB5, and HOXA6, and the differentially increased expression of lncRNAs that overlap SYT15 and SHANK2. Our results indicated a differential expression pattern for lncRNAs and targeted mRNAs in the peripheral lymphocytes in different neurological disorders.

Objective To investigate the interaction between the Nf1 (Neurofibromin 1)and the BDNF (brain derived neurotrophic factor) in the mechanism of attention deficit hyperactivity disorder (ADHD).Methods Spontaneously hypertensive rats were chosen as the experimental group and WKY servesd as control.RT-PCR,Western-blot were used to detect the mRNA and protein expression levels of Nf1 and BDNF.To explore their relationship,BDNF expression levels were detected after Nf1 expression plasmid transferred and transfection in PC12H and CBRH-7919 cells.Results were quantified by Image-pro plus software.Results In the PFC of ADHD model rats SHR,Nf1 expression was abnormally degressive(P =0.000),in common with the expression of BDNF(P =0.000).Ectopic expression of Nf1 further encouraged the expression of BDNF (PC12H:P =0.000,7919:P ＜0.05).Conclusion Nf1 expression was significantly lower in PFC of SHR than control,and positively correlated with the levels of BDNF.These findings show that in the prefrontal cortex,Nf1-BDNF dysregulation may be involved in the pathomechanism of ADHD.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">&lt;正&gt;缺血性脑血管病（脑缺血）是以脑循环血流量减少为特征的脑血管疾病,具有高发病率和死亡率,严重威胁人类健康。对于脑缺血防治药物的研究至今尚无突破,寻找新型脑缺血防治药物一直是国内外研究热点。脑缺血发生后,神经细胞发生程序性死亡,包括凋亡和自噬（autophagy）^[1]。近年来,自噬作用逐渐引起关注。胸腺素β4（thymosinβ4,Tβ4）是由43个氨基酸残基组成的多肽,属胸腺素β家族^[2],与新生血管发生、心肌修复</span>

Objective To evaluate vitamin D nutritional status and its relationship with ultrasonic bone mineral density among children in Wenzhou City.Methods Overnight fasting venous blood samples of 546 children were collected to measure the concentration of serum 25 -hydroxyvitamin D (25 (OH)D),and bone mineral density was measured.Results The average concentration of serum 25 (OH)D was 25. 42 ±0. 590 ng/mL,25. 09% of participants were found with sufficient vitamin D,the rate of severe deficiency,deficiency and insufficiency were 1. 47%,32. 60% and 40. 84%respectively.The concentration of 25 (OH)D in different age groups was statistically significantly different (P<0. 01 ). Vitamin D nutritional status of the older groups was poor than that of the younger ones.In the group of children aged 7 to 1 1 and 11 to 14,the levels of 25(OH)D of boy was significantly higher than that of girl.Compared to those with sufficient vitamin D,children with vitamin D deficiency or insufficiency had lower bone mineral density(BMD)(P<0. 05 ).The level of 25(OH)D was correlated with BMD (r=0. 197,P<0. 01).After adjusted age and BMI,there was still a positive correlation between 25(OH)D level and BMD (r=0. 115,P<0. 01).Conclusions The vitamin D level of children in Wenzhou area is low.The level of serum 25 (OH)D may be related to BMD .More attention should be paid to the supplementation of vitamin D.