Advances and Applications of Brain Organoids / 神经科学通报·英文版

Understanding the fundamental processes of human brain development and diseases is of great importance for our health. However, existing research models such as non-human primate and mouse models remain limited due to their developmental discrepancies compared with humans. Over the past years, an emerging model, the "brain organoid" integrated from human pluripotent stem cells, has been developed to mimic developmental processes of the human brain and disease-associated phenotypes to some extent, making it possible to better understand the complex structures and functions of the human brain. In this review, we summarize recent advances in brain organoid technologies and their applications in brain development and diseases, including neurodevelopmental, neurodegenerative, psychiatric diseases, and brain tumors. Finally, we also discuss current limitations and the potential of brain organoids.

Gestational hypothyroxinemia and offspring autism / 中华内分泌代谢杂志

Hypothyroxinemia is a common subclinical thyroid dysfunction in pregnant women. Epidemiological studies have shown that the offspring of maternal hypothyroxinemia have a significantly increased risk of autism, but mechanisms remain unclear. Studies from clinical and animals suggest that thyroid hormone deficiency may lead to irreversible damage to brain development, which may contribute to autism. This review explores the clinical evidence of gestational hypothyroxinemia and autism in offspring and the underlying mechanisms that promote the development and progression of autism. It also emphasizes the importance of early diagnosis and treatment of thyroid disease in pregnancy for the prevention of neurodevelopmental abnormalities in offspring.

Prenatal assessment of the cardiovascular structure and brain development of fetuses with tetralogy of Fallot on fetal MRI / 中华放射学杂志

Objective:To explore the value of prenatal MRI in assessing the cardiovascular structure and brain development of fetuses with tetralogy of Fallot (TOF).Methods:From August 2011 to April 2021, 30 fetuses of TOF with gestational age (GA) 18-33 (25±3) weeks were retrospectively enrolled in Shanghai Children′s Medical Center, School of Medicine, Shanghai Jiao Tong University. Sixty normal fetuses were selected with matched GA of TOF fetuses as control group. In 30 fetuses with TOF, the GA of 17 fetuses were less than 25 weeks and of 13 fetuses were more than 25 weeks. The balanced fast field echo sequence MRI was performed to measure the diameter of the aortic arch isthmus (AoI), the main pulmonary artery (MPA) and the ratio was calculated (MPA/AoI). The size of ventricular septal defect (VSD) was measured in TOF group. The single-shot turbo spin echo sequence MRI was performed to measure fetal brain biparietal diameter (BBD), brain frontal-occipital length (BFOL), skull biparietal diameter (SBD), skull fronto-occipital diameter (SOD) and head circumference (HC). The Pearson analysis was used to evaluate the correlation of the above indicators with GA. The independent sample t test was used to compare the differences in various indicators between TOF and control groups in both GA≤25 and GA>25 weeks. Results:The sizes of VSD in TOF fetuses were (3.5±1.1) mm (GA≤25 weeks) and (4.4±0.6) mm (GA>25 weeks). The AoI, MPA, BBD, BFOL, SBD, SOD, HC were positively associated with GA (TOF group: r=0.80, 0.50, 0.92, 0.93, 0.91, 0.84, 0.91, MPA P=0.005, others all P<0.001; control group: r=0.90, 0.87, 0.96, 0.95, 0.93, 0.95, 0.96, all P<0.001). For fetuses with GA≤25 weeks, there were 17 fetuses in TOF group and 34 fetuses in control group, and the AoI in TOF group was larger than that in control group ( t=10.00, P<0.001), the MPA, MPA/AoI, BFOL were smaller than those in control group (all P<0.05). No statistical significant differences of BBD, SBD, SOD, HC were found between the groups (all P>0.05), For fetuses with GA>25, there were 13 fetuses in TOF group and 26 fetuses in control group, and the AoI in TOF group was larger than that in control group ( t=3.36, P=0.002) and MPA, MPA/AoI, BBD, BFOL, SOD, HC were smaller than those in control group (all P<0.05). No statistically significant difference of SBD was found between groups ( P=0.068). Conclusions:MRI is feasible to measure the cardiovascular structure and the brain development of fetuses with TOF. Fetuses with TOF have varying degrees of VSD, MPA stenosis and AoI dilation. An altered brain growth of fetuses with TOF appears in the second trimester of pregnancy.

Anestesia general en infantes: consideraciones neurobiológicas y neuropsicológicas / General anesthesia in infants: neurobiological and neuropsychological concerns

Resumen La anestesia general es una herramienta imprescindible para el proceso quirúrgico, ya que disminuye el dolor, reduce la ansiedad y genera inconsciencia. Sin ella, las cirugías serían dolorosas, riesgosas y emocionalmente traumáticas. La reciente emisión de una alerta sobre el uso de fármacos anestésicos en niños menores de 3 años por parte de la Food and Drug Administration (FDA) de los Estados Unidos generó controversia en torno a sus posibles efectos negativos. En este artículo se abordan los principales hitos del desarrollo neurobiológico del niño y se revisan las posibles consecuencias neuropsicológicas del uso de anestesia general en esta población. La mayoría de los reportes que abordan este tema son de tipo retrospectivo y arrojan resultados controversiales por sus inherentes dificultades metodológicas. Sin embargo, el estudio prospectivo sobre seguridad del uso de anestesia general en niños de la Clínica Mayo (MASK, Mayo Anesthesia Safety in Kids), junto con otros estudios a gran escala, han confirmado algunos datos obtenidos en los estudios experimentales que dieron sustento a la alerta emitida por la FDA. Así, las evidencias hasta ahora publicadas sugieren que el uso de anestesia general es seguro para el desarrollo cognitivo general del niño, aunque evidencian también alteraciones focalizadas en procesos cognitivos específicos que deben ser consideradas por el médico y la familia ante un procedimiento quirúrgico-anestésico.

Abstract General anesthetics are crucial drugs for surgical interventions, which are indicated to induce analgesia, diminish pain, and reduce anxiety in order to facilitate invasive procedures. In pediatric patients, benefits of general anesthetics also include abolishment of motility. Besides their probed benefits on surgery, the recent warning of the Food and Drug Administration (FDA) on the use of general anesthetics in children yielded a controversy on their potential neurotoxic effects. In this review, the main facts of the cerebral development are studied, and the available evidence concerning the use of general anesthesia on the neuropsychological development of children is analyzed. Most of the studies found were uncontrolled retrospective cohorts for which conclusions are difficult to obtain. However, a few group of controlled studies, including the Mayo Anesthesia Safety in Kids study (MASK), have partially supported the FDA warning. Cumulated evidence appears to support the safety use of general anesthetics, but no conclusive data supporting that it may induce massive effects on the cognitive development of exposed children has been reported. Important evidence suggests that specific cognitive functions may result altered under long-term expositions. Such data must be considered for those involved in anesthetic procedures.

Rol de la osteocalcina más allá del hueso / Role of osteocalcin beyond bone

Los hallazgos osteológicos se intensi!caron en los últimos años. Se demostró que el esqueleto se comporta, además de sus funciones clásicas, como un órgano de secreción endocrina que sintetiza al menos dos hormonas: el factor de crecimiento de !broblastos 23 (FGF-23) y la osteocalcina (Ocn). La Ocn es un péptido pequeño que contiene 3 residuos de ácido glutámico. Estos residuos se carboxilan postraduccionalmente, quedando retenida en la matriz ósea. La forma decarboxilada en el primer residuo de ácido glutámico (GluOcn) fue reportada por poseer efectos biológicos; la resorción ósea es el mecanismo clave para su bioactivación. La presente revisión se centra en los conocimientos actuales sobre la función hormonal de la Ocn. A la fecha se reporta que la Ocn regularía el metabolismo energético aumentando la proliferación de células ` pancreáticas, y la secreción de insulina y de adiponectina. Sobre el músculo esquelético actuaría favoreciendo la absorción y el catabolismo de nutrientes. La función reproductiva masculina estaría regulada mediante el estímulo a las células de Leydig para sintetizar testosterona; en el desarrollo cerebral y la cognición, la Ocn aumentaría la síntesis de neurotransmisores monoaminados y disminuiría el neurotransmisor inhibidor GABA. Si bien son indispensables mayores evidencias para dilucidar los mecanismos reguladores por medio de los cuales actuaría la Ocn, los resultados enumerados en los distintos estudios experimentales establecen la importancia de este novedoso integrante molecular. Dilucidar su rol dentro de estos procesos interrelacionados en seres humanos abriría la posibilidad de utilizar a la Ocn en el tratamiento de enfermedades endocrino-metabólicas. (AU)

Osteological !ndings have intensi!ed in recent years. The skeleton behaves as an endocrine secretion organ that synthesizes at least two hormones: osteocalcin (Ocn) and !broblast growth factor 23 (FGF-23). Ocn is a small peptide that contains 3 glutamic acid residues. After translation, these residues are carboxylated to make possible its retention into the bone matrix. Decarboxylation on the !rst glutamic acid residue (GluOcn) has been reported to have biological effects. Bone resorption is the key mechanism for its bioactivation. This review focuses on current knowledge on Ocn hormonal function. It has been reported that Ocn regulates energy metabolism by increasing the proliferation of pancreatic ` cells, and the secretion of insulin and adiponectin. On the skeletal muscle, it may act by favoring the absorption and catabolism of nutrients. Male reproductive function might be regulated by stimulating Leydig cells to synthesize testosterone. Regarding brain development and cognition, Ocn would increase monoamine neurotransmitters synthesis and decrease inhibitory neurotransmitter GABA. Although more evidence is needed to elucidate the regulatory mechanisms of Ocn, different experimental studies establish the importance of this novel molecular mediator. Clarifying its role within interrelated processes in humans, might open the possibility of using Ocn in different treatments of endocrine-metabolic diseases. (AU)

Research progress in mechanisms underlying autism spectrum disorders / 中国实用儿科杂志

The autism spectrum disorders(ASDs)are a complex group of neuropsychiatric conditions defined by impairment in three core behavioral domains:social interaction,verbal and non-verbal communication,and restricted interests/repetitive behaviors. Extensive genetic studies have led to the identification of many autism susceptibility genes,and increased understanding on the contribution of de novo and inherited copy number variation. Here,we seek to place recent genetic findings within a developmental and brain circuit context,and approach the basic understanding of autism neuropathology from multiple genetic,molecular,cellular and neural circuit domains. The authors reviewed literatures that interrogates brain mechanisms of ASDs utilizing animal models,primarily in mice. Understanding genetic data within a brain development context will shed light on how individual risk genes operate to determine patient symptomatology,which will inform circuit specific behavioral interventions leading to better intervention and disease outcomes.

Effects of intrauterine exposure to thyroid dysfunction on the development of brain neurons in neonatal rats / 中华内分泌代谢杂志

Objective To investigate the expression of MCT8, DCX, SHH and ARC/ARG3. 1 in brain neurons of neonatal rats exposed to thyroid dysfunction in uterus. Methods Wistar pregnant rats were randomly divided into control group and experimental groups that rats were drunk water with 1, 3, or 5 ppm propylthiouracil ( PTU). The thyroid function and morphological changes of PND1 and PND7 were detected. The expression of MCT8, DCX, SHH, ARC/ARG3. 1 protein in cerebral cortex and hippocampus were detected by Western blot or immunohistochemistry. Results (1) The levels of TT4 decreased significantly in PND1 pups of PTU 3 ppm and 5 ppm groups (P＜0.05 or P＜0.01). The TSH levels significantly increased while FT4 levels significantly decreased in pups of PTU 5 ppm group on PND7 ( P＜0. 05). ( 2) The number NV, V, S, and cross-sectional area of thyroid follicles in offspring of PTU groups were significantly higher than those in the control group on postnatal day 1 and 7 (P＜0.05 or P＜0.01, respectively). (3) The expression of MCT8 in cortex and hippocampus gradually increased with the increase dose of PTU on two postnatal days, but there was significant change in PTU 5 ppm group on PND1 ( P＜0.05). The expression of SHH in pup cortex decreased with the increase of PTU exposure dose on PND7. DCX protein expression in the pup cortex on two postnatal days showed an uptrend with the increase of PTU exposure dose. ARC/ARG3.1 protein levels in hippocampal CA1 area of the pups increased significantly in PTU 1 ppm group on PND1 than that in the same-day control group ( P＜0. 05). Conclusion The damaged neurons of neonatal rats exposed to hypothyroidism in utero can be improved with the gradual recovery of thyroid function, but can not be completely restored to normal level.

Advance in MRI assessment of white matter injury and brain development in premature infants / 国际儿科学杂志

Premature white matter injury (WMI) is the immature white matter injury caused by ischemia,hypoxia and inflammation,which will affect normal neurodevelopment process and cause poor neurological outcomes.MRI has been widely used in early diagnosis of WMI because it can reflect the anatomical and pathological changes of WMI in premature infants.Functional MRI,such as diffusion-tensor imaging and magnetic resonance spectroscopy,can help quantitative assessment of the severity of WMI and neurodevelopment process so as to predict its neurodevelopment prognosis.This review summarizes the progresses in the application of MRI in the quantitative assessment of WMI and brain development in preture infants and its neurological outcomes.

Early repetitive pain in preterm infants in relation to brain development / 国际儿科学杂志

With the improvement of medical care the survival rate of preterm infants (gestational age ＜ 37 weeks) increased dramatically,however long-term neurodevelopmental and behavioral problems are still a concern.During their stay in neonatal intensive care unit preterm patients suffer lots of repeated painful procedures while it's a period of rapid brain development.Preterm infants have the nociceptive circuitry to feel the pain,but this system is functionally immature.Because the spinal neuromodulation system is immature that repeated pain stimulation leads to increased nociceptive signaling in the central nervous system.Besides,specific cell populations in the central nervous system of preterm infants are particularly vulnerable to oxidative stress and inflammation.Therefore,it is important that pain-related stress in preterm infants is appropriately managed.

Genes Biomarkers In Autism: Brain Development And Function

Myasthenia gravis is an autoimmune disorder that places patients in debilitating condition. It currently affects 14 to 20 per 100,000 population. Its pathogenesis involves the destruction of acetylcholine receptor by antibodies pro-duced by lymphocytes in the thymus gland. Symptoms could vary from impair extraocular muscles to generalized weakness. The antibodies have also been reported to affect other muscle structure within the body such as cardio-myocytes, leading to arrhythmia episodes which could be fatal. This review is a student project and involves the as-sessment of myasthenia gravis and the interplay between thymoma and thymomectomy.

La maduración cerebral en el niño: el caso de la adquisición del concepto de muerte y su evolución / Brain development in childhood: acquisition and evolution of children's understanding of death

Se presenta la relación entre la maduración cerebral en el niño y la adquisición de los cinco subcom ponentes biológicos del concepto maduro de la muerte: Irreversibilidad, aplicabilidad, inevitabilidad, cese funcional y causalidad. Las teorías actuales plantean que el conocimiento (entre otros el de la muerte) se realiza en un proceso de oleadas confluentes, con avances y retrocesos y esto explica la diferencia muy importante que se puede encontrar en niños de la misma edad y buen coeficiente in telectual. Generalizando, niños menores de 4 años no tienen conciencia de la muerte. Posteriormente descubren, con dificultad, el concepto de irreversibilidad y hay mucha discusión sobre el orden de adquisición del resto de los subcomponentes- lo que está relacionado con la incorporación del pen samiento dualístico y cierto grado de conocimiento biológico. No hay acuerdo sobre la influencia cultural, socio económica y experiencias previas con la muerte de familiares. Alrededor de los 12 años se considera que se logra un concepto maduro. Se revisa la incorporación de elementos sobrenaturales en la literatura infantil clásica y actual, comprobándose que su presencia es más marcada en los cuentos para preescolares occidentales que orientales en el siglo XXI.

There is a relation between brain development in children and the acquisition of the five subcom ponents for a mature conceptualisation of biological death: Irreversibility, applicability, inevitability, functional cessation and casuality. Current theories propose that knowledge (among them about death) is acquired through a series of confluent waves with advances and setbacks, thus explaining the important differences encountered among children of the same age and good intellectual develop ment. Generally speaking children under four years of age do not have a consciousness of death. They later discover, albeit with difficulty, the concept of irreversibility and much discussion exists about the order in which the remaining concepts are acquired-which is related to incorporating dualistic thought and a degree of biological understanding. There is no agreement on the influence culture, socioeconomic background and previous experience of deaths in the family may have. The mature concept is thought to be acquired around the age of 12. We look at the inclusion of supernatural ele ments in classical and current chidren's literature, proving that is more prevalent in western literature in the 21st century.

A review of effects of disrupted primary caregiver infant relationship on brain development and psychopathology / 中国心理卫生杂志

The disrupted caregiver infant relationships are not unpopular (infant abuse and non-traumatic caregiver-infant relationship).Animal and human studies have demonstrated this disruptive relationship exerts the fundamental and enduring impacts on stress system, limbic system and relevant cortex.Individual carrying such biological susceptibility might develop psychopathology under stresses at later life.The assessment needs to get involved the measures with more objectivity,real time and moment-to-moment components.The study on caregiverinfant relationship also should focus on the historical and cultural aspects of China.

Comparison of the cognitive function in rats at different brain developmental stages / 中国实验动物学报

Objective To investigate the relationship between different brain developmental stages and changes of cognitive function in rats. Methods 1-month, 2-month and 8-month-old rats were selected to imitate the juvenile, adolescent and adulthood, respectively, and their behavioral functions were compared. The reward operant conditioning and Morris water maze task were used to investigate the differences in exploration interest, executive and recognition ability, spatial learning and memory of the rats at different ages. Results In the reward operant conditioning and Morris water maze task, there was no significant difference in the cognitive function between 1-month and 2-month-old rats. In the reward conditioning phase, the nose pokes numbers of 8-month-old rats were significantly decreased compared with the 1-month-old rats (P＜ 0. 01). There was no significant difference in nose pokes accuracy. During the operant conditioning phase, the lever press numbers and accuracy of 8-month-old rats were significantly decreased ( P ＜ 0. 05 or P ＜ 0. 01) and the press latency was longer (P ＜ 0. 05). At the phase of visual identification, the press and reward numbers, and the visual identification index were significantly decreased ( P ＜ 0. 05 or P ＜ 0. 01). In the Morris water maze test, compared with the 1-month-old rats, the total swimming distance and escape latency of the 8-month-old rats were significantly increased (P＜ 0. 05), as well as average swimming speed ( P ＜ 0. 05 or P ＜ 0. 01) in spatial learning phage. In spatial memory phage, the swimming distance and time spent in the target quadrant were obviously decreased (P＜ 0. 01). Conclusions The cognitive functions of rats at different brain developmental stages are different. The juvenile and adolescent rats have similar cognitive functions, but 8-month-old adult rats appear decline in the exploration interest, executive and recognition ability, and spatial learning and memory function.

Effect of optimization of parenteral nutritional strategy on early brain development in premature infants / 中国新生儿科杂志

Objective To study the influence of optimization of parenteral nutrition strategy on the head circumference and brain volume in very low birth weight infants.Method Very low birth weight infants admitted to NICU of University of Hong Kong-Shen Zhen Hospital from January 2014 to June 2016 were assigned to optimized group and conventional group according to early nutritional strategies.Early parenteral nutrition intakes were increased in infants assigned to optimizated group.Nutrition intakes and parenteral nutrition related complications within 28 days after birth were compared between groups.All participants underwent brain MRI at corrected gestational age (CGA) 36 weeks.Head circumference and brain volume measured by MRI were also compared between groups.Result A total of 40 preterm infants were recruited,with 20 infants in each group.There were no significant differences in the gestational age,birth weight,brain injury and intrauterine growth retardation rate between the two groups (P ＞ 0.05).The average daily total calories and protein intake of optimization group during the first 4 weeks were significantly higher than those of conventional group,respectively [(101.5 ± 3.1) kcal/ (kg · d) vs.(96.1 ± 3.2)kcal/(kg·d),(3.07±0.16) g/(kg·d) vs.(2.90±0.11) g/(kg· d),P＜ 0.05].Theaverage daily calorie intake and protein intake of optimization group was increased by 4.7％ and 5.5％,compared with those of conventional group.Compared with the conventional group,head circumference and total brain volume of optimized group at CGA 36 weeks was improved by 3.3％ and 4.1％,and the differences were both statistically significant (P ＜ 0.05).Cerebral cortex gray matter volume of optimized group was significantly higher than that of conventional group [(102.4 ± 4.9) ml vs.(96.4 ± 4.6) ml,P ＜ 0.05].There was no significant difference in brain white matter,deep gray matter and cerebrospinal fluid volume between the two groups (P ＞ 0.05).Conclusion The optimization of parenteral nutrition within the framework of active nutrition strategy of preterm infants can further improve the early nutritional intake of preterm infants,leading to the increase of the head circumference and the gray matter volume of the cerebral cortex.

Development of extracerebral space with age in normal infants / 西安交通大学学报(医学版)

Objective To measure the extracerebral space width of normal infants with magnetic resonance imaging(MRI)and observe the variation tendency with age for quantitative diagnosis of hydrocephalus.Methods We measured the distance of interhemispheric fissure and bilateral frontal subarachnoid space on MRI images of 213 normal infants.The width of extracerebral space was expressed as mean ± standard deviation.The gender and bilateral differences were compared by rank sum test,and the variation trend of extracerebral space with age was observed by the scatterplots fitting regression curve.Results The width of infant extracerebral space had two milestones,4-month-old extracerebral space was the widest,and the amplitude of variation tended to be mild after 1 year of age.There was no significant difference between the genders in gap of extracerebral space(P＞ 0.05). However,bilateral frontal subarachnoid space differed significantly between genders(P＜0.05).Conclusion The width of extracerebral space,a dynamic process with age in infants,is most pronounced under 10 months old.The accurate diagnosis of pathological external hydrocephalus should be based on the reference value of the extracerebral space at different months,and it is very important to combine it closely with clinical history.

Assessment of neonatal brain development by 2D1H-MRS at 3.0T / 西安交通大学学报(医学版)

Objective To compare metabolite ratios in the basal ganglia regions between preterm and term neonates and analyze the correlation between metabolite ratio and postmenstrual age(PMA)using 2D magnetic resonance spectroscopy(MRS)at 3.0T,so as to explore the value of MRS in evaluating neonatal brain development.Methods Normal neonates who underwent 3.0T MRS exam ination were recruited.Axial 2D MRS was performed with the point-resolved spectroscopy sequence(PRESS)(echo time/repetition time,144 ms/1 000 ms) through the basal ganglia covering three regions of interest(ROI)(i.e.bilateral lenticular nucleus,thalamus and white matter beside the anterior horn of lateral ventricle).The peak areas ratios of Cho/Cr,NAA/Cho and NAA/Cr were calculated.Metabolite ratios were compared between preterm and term neonates,and between three ROIs. Correlation between metabolite ratio and PMA was further analyzed.Results Totally 54 neonates were included(preterm/term,27/27).In all the three ROIs,preterm neonates presented significantly higher Cho/Cr(P＜0.05) and significantly lower NAA/Cho ratios(P＜0.001)than those of term neonates;higher NAA/Cr ratio was only observed in the lenticular nucleus of preterm neonates(P＜0.001).Significant differences in metabolite ratios were detected among the three ROIs in preterm group(P＜0.05).For term group,metabolite ratios differed between lenticular nucleus and white matter beside the anterior horn of the lateral ventricle(P＜0.01),while no difference between lenticular nucleus and thalamus was found(P＞0.05).Significant correlation between metabolite ratio and PMA was found in all the three ROIs except NAA/Cr ratio in the white matter beside the anterior horn of the lateral ventricle.To be specific,we found linear negative correlation of Cho/Cr ratio(r= -0.325,-0.633,-0.438) and positive correlation of NAA/Cho ratio(r = 0.604,0.773,0.483)in all the three ROIs,but negative correlations of NAA/Cr ratio(r=0.487,0.367)in lenticular nucleus and thalamus with PMA.Conclusion Our findings suggest the tempo-spatial pattern of neonatal brain metabolite development in terms of anatomic location and PMA.Besides,the multi-voxel MRS presents potentials in establishing the normal topologic and age-matched reference in neonates and thus making brain maturation assessment and detection of early abnormal metabolism.

Advances in clinical applications of amplitude-integrated electroencephalogram in premature infants / 中华实用儿科临床杂志

Amplitude-integrated electroencephalogram(aEEG)is a new method for bedside monitoring of cerebral function which is gradually used in clinical routine.Although the grading criteria has not been unified, some studies suggest that aEEG recorded during the very early period after birth can not only indicate the maturation of brain development, but also provide values of clinical application on the early identification, determination of the severity and long-term prognosis assessment of brain injury(e.g.white matter injury, intracranial hemorrhage, etc.)in premature infants.

Atopic disease and attention deficit/hyperactivity disorder in children / 国际儿科学杂志

It is recently known that attention deficit/hyperactivity disorder(ADHD),which is often recognized in early-school children,may have a close relationship with atopic disease.Furthermore,there are numerous systemic reviews and meta-analysis focusing on the comorbidity of ADHD and atopic disease.However,the mechanism of the comorbidity has remained less well explored.This review aims to gain a better understanding of the relationship between ADHD and atopic disease in children,investigate the hypothesis and relevant studies results about the influences of atopic disease on ADHD,and help the correlative further research to some extent.

Construction and Validation of a Mammalian Expression Vector for in Utero Electroporation Study of miR-3099 in the Mouse Neocortex

@#Introduction: MiR-3099 was reported to play a role in neuronal cell differentiation/function in the brain during late embryonic and early neonatal development. To further explore its potential regulatory effects on embryonic brain development, this study aims to construct and validate an expression vector of miR-3099 for future gain-of-function and loss-of-function studies. Methods: pCAG-eGFP vector was modified to include IRES2 and miR-3099 with 150bp upstream and downstream genomic sequences. The newly constructed vector, pCAG-miR-3099-IRES2-eGFP, consists of CAG promoter. The in vitro expression level of miR-3099 was measured using stem-loop RT-qPCR after it was transfected into 293FT cell. Later, the vector was electroporated into the embryonic brain at E15.5. Three days later, the E18.5 embryonic brain was harvested and cryopreserved. Immunohistochemistry was performed by using antibody against eGFP to validate the in utero expression of the transgene in the neocortex of the brain. Results: Our finding showed that, the expression level of miR-3099 was significantly upregulated (p<0.001) in cells transfected with miR-3099 vector as compared to both negative and empty plasmid control groups. In addition, the expression of eGFP was noted in the brain section indicating that the vectors with or without miR-3099 transgene were successfully transfected into and expressed in the neocortex upon electroporation. Conclusion: The bicistronic expression vector of miR-3099 which was driven by the CAG promoter was successfully constructed, validated and sufficiently delivered to brain cells via the in utero electroporation approach. The regulatory roles of miR-3099 in embryonic brain development can be manipulated using similar approach.

Function of Znf804a in neuronal migration, proliferation and differentiation during mice brain development / 中国心理卫生杂志

Objective:To explore the function of Znf804a during brain development by using mouse model.Methods:The shRNA of Znf804a (shZnf804a) and control (pSUPER) plasmids were introduced into ventricular zone of ICR (Institute of Cancer Research) mice at E14.5 (three mice in each group) by using in utero electroporation.The speed of migration was evaluated by comparing the proportions of neuron in cortical plate (CP) zone.The proliferation speed was evaluated by comparing the diameters of neurospheres formed by neuron progenitor cells.The differentiation speed was evaluated by comparing the proportions of Nestin staining positive cells in neuron progenitor cells.Results:The proportion of neurons in CP zone was lower in shZnf804a group than in controls(11.8％ vs.75.4％,P ＜ 0.001).The diameter of neurospheres formed by neuron progenitor cells was bigger in shZnf804a group than in controls (295μm vs.172μm,P ＜0.01).The proportion of Nest in staining positive cells in neuron progenitor cells was larger in shZnf804a group than in controls (31.5％ vs.9.6％,P ＜0.01).Conclusion:It suggests that the migration speed of neurons in shZnf804a is lower than that in controls,the proliferation speed is higher than that in controls,and the differentiation speed is lower than that in controls.These results indicate that Znf804a may play an important role in the development of mouse brain.