OBJECTIVE: To analyze the clinical features and variants of ABCD1 gene in a Chinese pedigree affected with X-linked adrenoleukodystrophy. METHODS: Clinical data of the proband were collected and analyzed. Potential variant of the ABCD1 gene were analyzed by PCR and Sanger sequencing of the proband, his parents and 100 unrelated healthy individuals. RESULTS: The prominent features of the proband included cerebellar and brainstem lesions, along with increased serum level of very-long chain fatty acids. He was found to harbor a hemizygous c.1509delG (p.L504Sfs*54) variant of the ABCD1 gene, for which his mother was heterozygous. The same variant was not detected in his father and 100 healthy controls. CONCLUSION X-linked adrenoleukodystrophy has a variety of clinical manifestations. Discovery of the c.1509delG (p.L504Sfs*54), as a novel pathogenic variant of the ABCD1 gene, has enabled diagnosis and genetic counseling for this pedigree.
Adrenoleukodystrophy/genetics* ; Asians/genetics* ; China ; Female ; Genetic Testing ; Humans ; Male ; Mutation ; Pedigree

Objective:To investigate the differences in the expression profiles of cyclic RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of rheumatoid arthritis (RA) and its clinical significance.Methods:Venous blood were collected from 4 patients with RA (group T) and 4 healthy subjects (group C). The expression profiles of circRNA in PBMCs of the two groups were detected by Arraystar circRNA microarray, and the differentially expressed circRNA was analyzed by clustering analysis. The binding sites for interaction between differentially expressed circRNA and miRNA were predicted, and functional analysis such as geneontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed. quantitative real-time polymerase chain reaction (RT-qPCR) was used to verify the expression of partially differentially expressed circRNA in the two groups of PBMCs, and a circRNA-miRNA-mRNA regulatory network (ceRNA network) was constructed for the target circRNA with significantly differential expression. A receiver operating characteristic curve [receiver operating characteristic curve (ROC)] was established to analyze the potential diagnostic value of target circRNA. SPSS Statistics 23.0 and Graphpad Prism 8.0 were used to analyze the data, and the independent t test was used to analyze the difference between the two groups. Results:① Microarray results showed that, compared with group C, a total of 399 [fold of difference (FC)>1.5, and P<0.05] circRNA were abnormally expressed in PBMCs of group T; including 149 up-regulated and 250 down-regulated. ② Bioinformatics analysis: The prediction of the binding site of circRNA and miRNA suggested that the differentially expressed circRNA in RA might affect the inflammatory response by targeting miR-140-5p, miR-338-5p, and miR-9-5p. GO analysis showed that the differentially expressed circRNA was mainly involved in the intimal-binding organelles, protein metabolism and binding, etc. KEGG pathway analysis showed that most of the involved pathways were related to infection and human immune dysregulation. ③ The results of multi-sample RT-qPCR validation showed that the expression level of hsa_circRNA_009012 in group T was significantly higher than that in group C ( t=-4.417, P<0.01), the expression level of hsa_circRNA_101328 was significantly lower than that in group C ( t=-1.042, P<0.01), and the expression of hsa_circRNA_058230 had no significant change ( t=4.691, P>0.05). ④ ROC curve analysis indicated that hsa_circRNA_009012 had potential value in the diagnosis of RA [area under curve=0.96]. Conclusion:The expression of circRNA in PBMCs of patients with RA is imbalanced, and it may participate in the regulation of the development of RA. Among them, hsa_circRNA_009012 is expected to become a new biological marker for the diagnosis and treatment of RA.

The development of a cerebral organoid culture in vitro offers an opportunity to generate human brain-like organs to investigate mechanisms of human disease that are specific to the neurogenesis of radial glial (RG) and outer radial glial (oRG) cells in the ventricular zone (VZ) and subventricular zone (SVZ) of the developing neocortex. Modeling neuronal progenitors and the organization that produces mature subcortical neuron subtypes during early stages of development is essential for studying human brain developmental diseases. Several previous efforts have shown to grow neural organoid in culture dishes successfully, however we demonstrate a new paradigm that recapitulates neocortical development process with VZ, OSVZ formation and the lamination organization of cortical layer structure. In addition, using patient-specific induced pluripotent stem cells (iPSCs) with dysfunction of the Aspm gene from a primary microcephaly patient, we demonstrate neurogenesis defects result in defective neuronal activity in patient organoids, suggesting a new strategy to study human developmental diseases in central nerve system.
Action Potentials ; physiology ; Biomarkers ; metabolism ; Cell Culture Techniques ; Embryoid Bodies ; cytology ; metabolism ; Gene Expression ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; Lateral Ventricles ; cytology ; growth & development ; metabolism ; Microcephaly ; genetics ; metabolism ; pathology ; Models, Biological ; Mutation ; Neocortex ; cytology ; growth & development ; metabolism ; Nerve Tissue Proteins ; deficiency ; genetics ; Neurogenesis ; genetics ; Neurons ; cytology ; metabolism ; Organoids ; cytology ; metabolism ; PAX6 Transcription Factor ; genetics ; metabolism ; Patch-Clamp Techniques ; SOXB1 Transcription Factors ; genetics ; metabolism ; Zonula Occludens-1 Protein ; genetics ; metabolism

The morphology of the GnRH neurons in the rat brain during postnatal development was studied quantitatively using the ABC immunostaining method and an image analyser. The pattern of distribution of the GnRH neurons in different ages animals used was similar. GnRH cells of rats aged 1-7 days were small in size with short processes, pale-stained, and smooth in outline The number of GnRH cells in 1 day rats was not significantly different from that of adult, but a significant decrease in the GnRH cell number appeared in 7 day rats. In postnatal day 14, Gn-RH neurons reached adult level in cell body size, number and staining intensity. The GnRH fibers and terminals in the median eminence were gradually increased and reached adult level up to 14 postnatal days. These results suggested that the number of GnRH neurons is determined shortly after birth; the second postnatal week is the critical period for the development of GnRH neuronal morphology; and the establishment of function of the GnRH neuronal system may be earlier than its morphological maturation.

The distribution relationships of gonadotropin-releasing hormone (GnRH) (5-HT), tyrosine hydroxylase (TH), neurophysin (NP) and ?-endorphin (?-END) in the diagonal band of Broca (DBB) of rat brain were studied by immunohis- tochemistry. The results indicate that there was a close spatial overlap in the distribution between 5HT,TH,NP,?-END fibers and GnRH perikarya and fibers in the ventromedial area of VDBB and dorsomedial area of HDBB. In the dorsomedial area of VDBB close overlap in the distribution between 5-HT fibers and GnRH neurons was also observed. In addition, some TH and NP immunoreactive neurons were found in the medial areas of VDBB and HDBB, The close relationships in distribution and functional significance of GnRH,5-HT, TH, NP and ?-END were discussed.

The antifertility and antinidation effects of three extracts (benzol, ethyl acetate and alcohol) of Cyathula Oficinalis Kuan on mice were observed. The results indicate that the benzol extract had significant antifertility action. Its effective rate of antifertility was 100%, when 250 mg/kg was given orally. As compared with control, when 250 mg/kg ethyl acetate extract was given, the rate of antifertility was 63.7%. If 500 mg/kg ethyl acetate extract was given the effective rate of antifertility increased over 90%. However, when 250～500 mg/kg alcohol extract was given, the rate of antifertility was only 40～54%. The study of antinidation showed that the benzol extract (500 mg/kg )could effectively prevent implatation. The rate of antinidation was 100%, and yet the ethyl acetate and alcohol extracts were 66.7% and 40% repectively. The results suggest that the three extracts of Cyathula Officinalis Kuan all had antifertility effect, but the action of benzol extract was more effective than other two extracts.

By means of chromalum-hematoxylin and thiosulfate aldehydefuchsin stains the magnocellular neurosecretory neuron of the rat hypothalamus was studied. The cell body of neuron containing various amounts of secretory granules was large in size, with a variety of shapes. The axon of neuron showed an beaded appearance; only one in each neuron was very fine in diameter and left hypothalamic nuclei to form the hypothalamo-neurohypophyseal tract. The dendrite, in general, thicker than the axon, did not project outside of the nucleus. Many of the thick short dendrites did not contain secretory granules or only few secretory granules. The thick processes with more secretory granules were considered as dendritic processes. A number of axons and thick processes containing secretory granules also contacted with the endothelium of vessels and the ependyma of the third ventricle. In addition, a group of magnocellular secretory neurons were found in the sub-choroid plexuses of the lateral ventricle.

By means of ABC immunoenzyme technique, the distribution of GnRH neurons and fibers in the brains of rats and mice was observed, The results showed that more than 90% of GnRH neuronal perikarya were concentrated in the diagonal band-medial preoptic area near the organum vasculosum lamina terminalis, extending rostro-dorsally to the medial septic nucleus and caudo-laterally to the supraoptic nucleus region. No GnRH cell body could be found in the mediobasal hypothalamus, GnRH fibers terminated in the median eminence via the periventricular and lateral tracts. In the rostral median eminence, fibers were present in its entire layers and width, and caudally separated into two laterally-located bundles. The difference of GnRH system between rodents and primates, as well as its significance in reproduction are discussed.

Observations of the ultrastructural changes in neurons of hypothalamie arcuate, supraoptic and paraventricular nuclei were carried out in the rats following gossypol treatment at a dosage of 30mg/kg/day for 5 weeks. The results showed that after gossypol treatment no marked changes in supraoptic and paraventricular cells could be seen at electron microscopic level, and that there appeared a series of ultrastrucrural changes reflecting increased neurosecretory activity in some of arcuate neurons.They were as follows: Golgi complex and rough endoplasmic reticulum were slightlydilated and filled with homogeneous, low electron density materials in their cisternae; the amounts of neurosecretory granules, lysosomes, nucleolus-like bodies, and microtubules were increased. The nature, significance and cause of these changes are discussed.

Gonadotropin-releasing hormone(GnRH)prohormone of human and rat consistsof GnRH and GnRH-associated peptide(GAP).In this study,three antisera againstN-terminal,mid-region and C-terminal of GAP,and ABC immunoenzyme methodwere used to observe the GAP neurons in the brains of the rat,mouse and guineapig.The distribution of GAP neurons in these animals was similar.GAP perikaryawere mainly present in the septo-preoptic area,with the largest concentration inthe diagonal band near the organum vasculosum of the lamina terminalis.SomeGAP perikarya were also seen in the brain area near the supraoptic nucleus.GAPfibers were widely present in the forebrain and hypothalamus,and terminated inthe organum vasculosum of lamina terminalis and median eminence.Of 3 GAP antisera,the one against N-terminal gave more immunoreactive elements and moreintense staining.The morphology and distribution of GAP perikarya,fibers andterminals were similar to those of GnRH.These results,combined with other relatedfacts,suggest that there is a common GnRH prohormone in mammals,and itsprocessing products,GAP(or cleavage fragments)and GnRH,are cosecreted intohypophysial portal system to regulate anterior pituitary hormone secretion.