Huperzine A injection ameliorates motor and cognitive abnormalities via regulating multiple pathways in a murine model of Parkinson's disease.

As a common progressive neurodegenerative disorder, the satisfied therapies for Parkinson's disease (PD) are still unavailable. As a natural acetylcholinesterase inhibitor, the neuroprotective characteristic of Huperzine A (HupA) was supported by previous studies. However, questions remain on whether HupA injection (HAI, a main preparation of HupA) intervention conduces to PD treatment and if so, the potential molecular mechanisms. In this study, the efficacies of HAI treatment on PD-like pathological phenotypes were evaluated in a MPTP-induced PD murine model. The network pharmacology, transcriptome sequencing and experimental verification were integrated to comprehensively reveal the primary molecular mechanisms. Therapeutically, HAI intervention significantly improved the impaired locomotor behaviors as well as learning and memory abilities, and prevented the degeneration of dopaminergic neurons of PD mice. The network pharmacology analysis combined with experimental results showed that HAI treatment could effectively restore the disordered transcriptional levels of inflammatory factors and apoptosis related genes in the SNpc and striatum tissues of PD mice. Transcriptome sequencing results found that inflammation and oxidative phosphorylation served as significant functional mechanisms involved in HAI administration. The experimental verification indicated that HAI treatment effectively regulated the abnormal transcription levels of inflammation and oxidative phosphorylation related hub genes in the hippocampal samples of PD mice. In addition, molecular docking suggested strong affinity between HupA and the above core targets. Overall, this work displayed the reliable therapeutic effects of HAI on ameliorating the pathological symptoms of PD mice via modulating multiple pathways. The current findings were expected to provide a potential anti-PD agent.

Expression and potential role of FOSB in glioma.

Background: FOSB is reported to be an oncogene in a variety of tumors. However, the expression and role of FOSB in glioma remain obscure. In this study, we aimed to explore the expression of FOSB in glioma and its biological role in glioblastoma multiforme (GBM). Methods: Western blot, immunohistochemical staining, and quantitative real-time polymerase chain reaction (RT-qPCR) were used to detect the expression of FOSB in clinical samples. FOSB was knocked down in cells to determine the effects of FOSB on the phenotypic changes of tumors by plate cloning, CCK-8 assay, and Transwell assay. Finally, subcutaneous tumorigenesis in nude mice was used to observe the tumorigenesis of glioma cell lines after the knockdown of the FOSB gene. Results: FOSB expression was higher in glioma compared with normal brain tissue. After the downregulation of FOSB, the expression of cleaved caspase-3 increased. Plate cloning and CCK-8 experiments showed that the proliferation of glioma cell lines decreased. The Transwell assay demonstrated that the glioblastoma cell lines had lower migration ability after the knockdown of FOSB. Finally, the tumor volume of U87 glioma cells in group sh-FOSB was smaller than that in the control group. The TUNEL staining in vitro showed that the apoptosis of sh-FOSB glioma cells increased. Conclusion: FOSB was highly expressed in glioma tissues. The viability of glioma cells decreased, and the ability of glioma cells to proliferate and migrate was reduced when FOSB was downregulated. Hence, FOSB may promote the development and migration of gliomas.

Expression of FOXO transcription factors in the brain following traumatic brain injury.

Traumatic brain injury (TBI) is a substantial clinical and social problem worldwide, causing high morbidity and mortality along with significant economic and medical costs. Forkhead box O transcription factors (FOXOs) have been found to play a critical role in the regulation of cell functions, such as nutrient metabolism, programmed cell death, and tumor suppression. In the central nervous system, FOXOs are reported to be pivotal regulators of learning and memory, neurite outgrowth, and axonal degeneration. However, the role of FOXOs in TBI is still unknown. Here, we investigate changes in the expression of FOXOs in the acute stage following TBI. First, we evaluated the expression of FOXO proteins in the brains of humans after TBI. A TBI model was then established in mice, and the ipsilateral cerebral cortex was collected at 3 h, 6 h, 9 h, 12 h, 24 h, and 72 h post-TBI. The dynamic expression of Foxo proteins was observed. Neuron-specific localization of Foxos was detected by double immunofluorescence staining. Following TBI, FOXO proteins in the brains of humans were significantly increased. In mice, Foxo protein levels generally peaked at 24 h. By examining co-localization with neurons, the proportion of Foxo(+) neurons was found to increase following TBI and peak at 24 h. This study reveals the time-dependent and neuron-specific expression of Foxos following TBI in mice, providing insight to enhance understanding of the role of Foxos in TBI.

Management of a "suspected ward" in a COVID-19 designated hospital in Wuhan.

During December 2019, an outbreak of unexplained pneumonia occurred in Wuhan, Hubei Province. The disease was subsequently named coronavirus disease 2019 (COVID-19) and the causative virus as severe acute respiratory syndrome conronavirus-2 (SARS-CoV-2). Based on experience, it is vital to exclude or diagnose suspected patients as soon as possible to prevent disease spread. Our hospital is a COVID-19 designated hospital in Wuhan. During the epidemic period, there was a reconstruction of the medical facilities to accommodate patients with different disease status. We document the development of "suspected ward," a ward that cared for patients with suspected COVID-19, in a large designated hospital during the COVID-19 outbreak in Wuhan City, China, and explain the suspected ward spatial layout, organization structure, diagnosis, and treatment flow chart of suspected cases. The key characteristics of our "suspected ward" is isolation, triage, fast diagnosis, and rapid referral. Our description of this suspected ward provides a reference for further improvements in the care of patients with suspected disease in emergency medical institutions.

STIM1 controls calcineurin/Akt/mTOR/NFATC2-mediated osteoclastogenesis induced by RANKL/M-CSF.

Store-operated Ca2+ entry (SOCE) is the stable calcium channel influx in most cells. It consists of the cytoplasmic ion channel ORAI and endoplasmic reticulum receptor stromal interaction molecule 1 (STIM1). Abolition of SOCE function due to ORAI1 and STIM1 gene defects may cause non-perspiration, ectoderm dysplasia and skeletal malformations with severe combined immunodeficiency (CID). Calcineurin/mammalian target of rapamycin (mTOR)/nuclear factor of activated T cells 2 (NFATC2) is an important signalling cascade for osteoclast development. Calcineurin is activated by Ca2+ via SOCE during osteoclastogenesis, which is induced by receptor activator of NF-κB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). However, the underlying mechanism has remained to be fully elucidated, which was therefore the aim of the present study. In the current study, flow cytometry was used to examine the effect of a number of STIM1 mutations on proliferation, differentiation, and expression of osteolysis-associated proteins in Bone marrow-derived mononuclear macrophages (BMDM). The calcineurin/AKT/mTOR/NFATC2 signaling cascade activation were also assessed. BMDMs were obtained from three patients with STIM1 mutations (p.E136X, p.R429C and p.R304W). These mutations, which exhibited abolished (p.E136X, p.R429C) or constitutively activated (p.R304W) SOCE, failed to respond to RANKL/M-CSF-mediated induction of normal osteoclastogenesis. In addition, activation of the calcineurin/Akt/mTOR/NFATC2 signalling cascade induced by RANKL/M-CSF was abnormal in the BMDMs with STIM1 mutants compared with that in BMDMs from healthy subjects. In addition, overexpression of wild-type STIM1 restored SOCE in p.R429C- and p.E136X-mutant BMDMs, but not in p.R304W-mutant BMDMs. Of note, calcineurin, cyclosporin A, mTOR inhibitor rapamycin and NFATC2-specific small interfering RNA restored the function of SOCE in p.R304W-mutant BMDMs. The present study suggests a role for SOCE in calcineurin/Akt/mTOR/NFATC2-mediated osteoclast proliferation, differentiation and function.

FOXO4 expression associates with glioblastoma development and FOXO4 expression inhibits cell malignant phenotypes in vitro and in vivo.

BACKGROUND AND AIM: Forkhead box protein O4 (FOXO4) is a transcription factor, and aberrant FOXO4 expression is associated with development of various human cancers. This study explored the role of FOXO4 in glioma in vitro and in vivo. METHODS: FOXO4 expression was first assessed in normal brain tissues, low-grade glioma, glioblastoma multiforme (GBM), normal human astrocytes (HA), and GBM cell lines, while manipulation of FOXO4 expression in glioma cell lines was assessed using qRT-PCR, Western blot, and cell viability CCK-8, Transwell, and a nude mouse subcutaneous xenograft assays. KEY FINDINGS: The data showed downregulated FOXO4 expression in GBM tissues and cell lines. FOXO4 overexpression induced by transfection with FOXO4 cDNA significantly inhibited GBM cell proliferation, migration, and invasion, but increased tumor cells to undergo apoptosis in vitro, while suppressed growth of GBM cell subcutaneous xenografts in nude mice. In conclusion, FOXO4 possesses an anti-cancer glioma activity, which could be a novel target for future control of GBM.

[Morphological study on early development of brain derived neurophic factor-positive neurons in the frontal lobe of human fetus].

OBJECTIVE: To investigate the growth and development of brain derived neurophic factor(BDNF)-positive neurons in the frontal lobe of human fetus. METHODS: The expression of the BDNF-positive neurons in the frontal lobe of human fetus in the 2(nd),3(rd),and 4(th) month of gestation were observed with the streptavidin-biotin-complex/immunoperoxidase(SABC)method. RESULTS: By the second month of gestation,BDNF-positive neurons were seen in the subventricular layer of the frontal lobe of cerebellum.By the third month of gestation,BDNF-positive neurons in the central layer were in various shapes,with big nucleus,less cytoplasm,and small processes.By the fourth month of gestation,BDNF-positive neurons in the central layer grew larger in size,cytoplasm increased,the BDNF-positive expression was enhanced with deeper dyeing,and the nerve fibers and particles were distributed between neurons;also,the BDNF-positive neurons were seen in the marginal layer of the frontal lobe of cerebrum. CONCLUSION: BDNF-positive neurons may participate in the early development of the frontal lobe of cerebrum of human fetus.

Temporal profile of nerve growth factor expression in the partial central nervous system of the Yangtze alligator Alligator sinensis (Reptilia,Crocodylia) during early postnatal growth.

Expression of nerve growth factor (NGF) in structures of the partial central nervous system of the Yangtze alligator, Alligator sinensis (Reptilia, Crocodylia) was examined during early postnatal growth using immunohistochemistry and Western blot assays. In animals 0-2 years of age NGF-positive cells in the cerebral cortex increased gradually in number and size, and were predominantly distributed in the molecular layer. NGF-positive cells in the midbrain showed similar increases but with predominant distribution in the ependymal layer. NGF-positive cells increased in the cerebellum between 0 and 1 years of age, with increased NGF expression being seen during the first 2 years of life mostly in the ependymal layer. NGF-positive cells were mainly found in the gray matter of the spinal cord with decreasing cell numbers, NGF expression levels being seen from 0 to 2 years and small processes without synaptic connection from 1 to 2 years. These results suggest that NGF is involved in the early postnatal growth of several structures of Yangtze alligator partial central nervous system, suggesting a possible role of NGF in the Yangtze alligator partial central nervous system.

[Morphological study on development of nerve growth factor-positive neurons in the cerebellum of human fetus].

OBJECTIVE: To investigate the growth and development of nerve growth factor (NGF)-positive neurons in the cerebellum of midanaphase human fetus. METHODS: The expression of the NGF-positive neurons in the cerebrum of human fetus was observed by immunohistochemical methods, and the integral absorbance (IA) was detected. RESULTS: By the 3rd to 4th month of gestation, neurons was seen in the ependymal, central, and marginal plate of cerebellum; the nucleus was oval and the neurons had short and small processes. By the 5th to 7th month of gestation, the number of NGF-positive neurons increased, the expressions enhanced, the nucleus was round-, oval-, or fusiform-shaped, the neurons grew larger in size, and the Purkinje cells showed NGF-positive expression. By the 8th to 10th month of gestation, the NGF-positive expression was enhanced with deeper dying, the body of Purkinje cells grew larger gradually, and the number of NGF-positive neurons in the granular cell layer and molecular layer increased. IA of the cerebellar cortical neurons of the 3rd, 4th, 5th, 6th, 7th, and 8th month of gestation showed an increasing trend, and significant difference was observed (P < 0. 05). CONCLUSION: NGF-positive neurons in the cerebellum play an important role for differentiation, proliferation, migration, and growth of neurons in the cerebellum.

[Morphology of nitric oxide synthase positive neurons in the cerebellar cortex of midanaphase human fetus].

OBJECTIVE: To investigate the growth and development of nitric oxide synthase (NOS)-positive neurons in the cerebellum of human fetus in the midanaphase. METHOD: The positive expression of the NOS-positive neurons in the cerebellum of midanaphase human fetus was observed by immunohistochemistry. RESULTS: By the sixth to seventh month of gestation, NOS-positive neurons were seen in the ependymal layer of the cerebellum. The nucleus was oval-shaped and the neurons had short and small processes. By the eighth to ninth month, NOS-positive neurons were found in the central layer of the cerebellum and the nucleus was round-, oval-, or fusiform-shaped; meanwhile, the neurons grew larger in size with richer cytoplast and heavier staining. The beaded nerve fibers reached the marginal layer and the layer became thickened on the tenth month, which generally was composed of 5 to 6 layers of NOS-positive neurons that were tightly aligned. Some NOS-positive neurons were in smaller size with the cell body and the nerve fibers grew well. CONCLUSION: Nitric oxide generated by NOS of the NOS-positive neurons in the cerebellum plays an important role in the differentiation, proliferation, and migration of neurons and gliacytes.

[Immunohistochemical study on development of nitric oxide synthase-positive neurons in the frontal lobe of cerebrum of midanaphase human fetus].

OBJECTIVE: To investigate the development of nitric oxide synthase (NOS)-positive neurons in the frontal lobe of the cerebrum of human fetus in midanaphase. METHODS: The positive expression of the NOS-positive neurons in the frontal lobe of cerebrum of human fetus was observed by immunohistochemistry. RESULTS: By the 7th to 8th month of gestation, NOS-positive neurons in the cortical plate of frontal lobe demonstrated themselves inequality of sizes and morphological difference in the deeper layers with interspersed distribution and increased NOS response, and the distribution of beaded nerve fiber was observed between neurons of cerebral tissues. By the 9th to 10th month of gestation, NOS-positive neurons in the deeper layers of cortical plate of frontal lobe developed slightly in size of the cell body with richer cytoplast, full shape and deeper dyeing and extrusive beaded nerve fibers, and the NOS-positive neurons scattered in the shallow layer of cortical plate presented with round or oval shape. The nucleus developed bigger but with sparse cytoplasm and clear nerve process. CONCLUSION: NOS-positive neurons in the deeper layer of cortical plate of lobus frontal consist of largely network of neural system and produce micro-environment with higher concentration of NO, which favors the differentiation, proliferation, migration, and development of various neurons.