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Objective To study the effect of ghrelin gene knockout on the postsynaptic potential of dopaminergic neurons in the substantia nigra of mice. Methods Substantia nigra tissue was taken from 10-week-old ghrelin-/- male mice (ghrelin-/- group) and their male wild-type (WT) littermates (WT group). RNA-seq technique was used to screen differently expressed genes (DEGs). KEGG pathway enrichment analysis was performed for the neuronal synaptic activity-associated signaling pathways which DEGs might be involved in. Quantitative real-time PCR (RT-qPCR) was used for validation of the identified DEGs, and western blotting was used for protein expression of neuronal synaptic activity-related genes. Results Compared with the WT group, the ghrelin-/- group had significant changes in the expression levels of 23 genes in the synaptic signaling pathway of dopaminergic neurons. Specifically, Ionotropic glutamate receptors α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid type subunit 3 (GluA3) and glycogen synthase kinase3-β (GSK-3β) regulated α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor and N-methyl-D-aspartic acid receptor on the postsynaptic membrane of neurons, respectively. GluA3 and GSK-3β genes were significantly downregulated in the substantia nigra tissue of mice in the ghrelin-/- group. RT-qPCR results showed that compared to the WT group, the ghrelin-/- group had significant downregulation of the mRNA expression levels of GluA3 and GSK-3β in the substantia nigra tissue of mice (t =2.408,2.740,P

Objective To investigate the effect of OTUD3 knockout on the expression of connexin 43 (Cx43) in neural stem cells (NSCs). Methods Brain cortex tissue protein was isolated and extracted from wild-type (WT) and OTUD3-/- embryonic mice on day 14 of gestation, and WT-NSCs and OTUD3-/--NSCs were cultured in vitro and subcultured according to cell status. After subculture to the third generation, an inverted microscope was used to observe the growth state of OTUD3-/--NSCs and WT-NSCs on days 1 and 3. Immunofluorescent staining was used to measure the expression of Ki67, Nestin, and β-tubulin Ⅲ in WT-NSCs and OTUD3-/--NSCs, and Western blotting was used to measure the changes in the expression of OTUD3 and Cx43 in WT-NSCs,OTUD3-/--NSCs, and the cerebral cortex of WT and OTUD3-/- embryonic mice. Results WT-NSCs and OTUD3-/--NSCs cultured and subcultured to the third generation showed a good growth status, and immunofluorescent staining showed positive staining of Ki67, Nestin, and β-tubulin Ⅲ. Compared with WT-NSCs,OTUD3-/--NSCs had significant reductions in the expression levels of OTUD3 and Cx43 (t=16.82,3.21,P

Objective To investigate the differentially expressed genes (DEGs) in the substantia nigra of mice with Parkinson’s disease (PD) induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) based on bioinformatics, as well as their role in the pathological and progression of PD. Methods GEO database was used to obtain the gene expression microarray data in the substantia nigra of mice in the normal group and the PD model group, and the bioinformatics method was used to screen for DEGs in the substantia nigra between the normal group and the PD model. Metascape tool was used to perform GO functional enrichment analysis and KEGG pathway enrichment analysis. Quantitative real-time PCR was used to measure the mRNA expression levels of key functional genes regulating neuronal apoptosis and prion transmission. Results There were 174 upregulated DEGs and 173 downregulated DEGs in the substantia nigra of mice with MPTP-induced PD (P0.26). The upregulated genes were significantly enriched in the pathways regulating prion transmission, neuronal development, iron homeostasis, and export across the plasma membrane, while the downregulated genes were significantly enriched in the pathways regulating synaptic vesicle release, transsynaptic complex transmission, neuronal apoptosis, and actin in cytoskeleton. Compared with the normal group, the PD model group had significant reductions in the expressions of Bdnf and Fbxw7 in the substantia nigra of mice (t=2.25,2.39,P0.05). Conclusion There are significant changes in the key functional genes Bdnf and Fbxw7 that regulate neuronal apoptosis in PD model, which lays a foundation for further clarifying their role in the pathological progression of PD.

Objective To explore the effect of hypoxia on the proliferation, differentiation and apoptosis of neural stem cells (NSCs) isolated from adult rat subventricular zone (SVZ) and cultured in vitro. Methods The NSCs isolated from adult rat SVZ were cultured under normoxia condition or hypoxia condition (hypoxia for 6, 12, 24, 48 or 72 h) in vitro. The nestin+ celis, Tju-1+ celis, GFAP+ cells and 04+ celis were detected by immunofluorescence staining, and the percentage of the positive cells was measured respectively. The proliferation of the NSCs was assayed with cell proliferation reagent (WST-1) and the cell cycle was detected by flow cytometry (FCM) . The apoptosis of the NSCs was detected by TUNEL and flow cytometry. Results The NSCs from adult rat SVZ were successfully isolated and cultured in vitro. Induced by serum, the NSCs differentiated at the 4th generation and most of them were Tju-1-positive (53.8±6.8%) whlle some of them were GFAP-positive (21.6±5.4%). Betides, a small number of them were O4-positive (11.4±4.5%). Hypoxia (0.3% O2) for a short peiiod (<24 h) promoted the proliferation of the NSCs (P<0. 05). But prolonged hypoxia (>24 h) reduced the proliferation capacity of the NSCs and increased the percentage of apoptotic cells (P<0 .01). Conclusion NSCs are present in normal adult tissues; NSCs isolated and cultured in vitro can differentiate into various types of cells. Shot term hypoxia can promote the proliferation of NSCs, but prolonged hypoxia induces apoptosis of NSCs. [ABSTRACT FROM AUTHOR]

Objective: To analyze and summarize the clinical efficacy and safety of lymphocyte apheresis combined with plasma exchange in the treatment of patients with hepatitis B virus-related liver failure at the ascending stage., Methods: A observational study was conducted. A total of 69 hepatitis B virus-related liver failure at the ascending stage patients who were hospitalized at Affiliated Guangzhou Eighth People's Hospital of Guangzhou Medical University from January 2016 to December 2020 were enrolled in this study. The patients were grouped according to their condition and wishes, including 38 patients treated with conservative medical treatment (control group) and 31 patients treated with lymphocyte apheresis combined with plasma exchange based on comprehensive medical treatment (study group). Clinical data were compared between the two groups 1-4 weeks after treatment, including dynamic changes of total bilirubin (TBil), international normalized ratio (INR), alanine aminotransferase (ALT), model for end-stage liver disease (MELD) score, and the rate of clinical improvement at 4 weeks after treatment. In addition, the adverse effects and dynamic changes of white blood cell count (WBC), lymphocyte count (LYM), platelet count (PLT), and hemoglobin (Hb) within 4 weeks after treatment were compared between the two groups., Results: Both groups showed significant improvement in clinical parameters after 1-4 weeks of initiation of therapy. The improvement of TBil, INR and MELD score at 1-4 weeks after treatment were significantly better in the treatment group than those in the control group [TBil (μmol/L): 248 (117, 335) vs. 398 (328, 464) at 1 week, 173 (116, 278) vs. 326 (184, 476) at 2 weeks, 107 (84, 235) vs. 355 (129, 467) at 3 weeks, 70 (61, 172) vs. 290 (82, 534) at 4 weeks; INR: 1.72±0.70 vs. 2.13±0.69 at 1 week, 1.67±0.61 vs. 2.28±1.35 at 2 weeks, 1.65±0.75 vs. 2.15±0.92 at 3 weeks, 1.61±0.93 vs. 2.19±1.17 at 4 weeks; MELD score: 18.35±5.32 vs. 23.38±4.56 at 1 week, 16.47±5.16 vs. 23.71±7.94 at 2 weeks, 16.30±5.75 vs. 22.64±6.99 at 3 weeks, 14.63±6.76 vs. 20.97±8.19 at 4 weeks], with significant differences (all P < 0.05). In addition, ALT levels at 1 week and 2 weeks after treatment in the study group were significantly lower than those in the control group [U/L: 128 (93, 206) vs. 240 (167, 436) at 1 week, 64 (42, 110) vs. 85 (69, 143) at 2 weeks, both P < 0.05]. The rate of clinical improvement at 4 weeks after treatment in the study group was 54.84% (17/31), which was significantly higher than that in the control group [28.95% (11/38)], with statistically significant difference (P < 0.05). There was no significant difference in the rate of new infection between the study group and the control group [22.58% (7/31) vs. 34.21% (13/38), P > 0.05]. Additionally, expect that the PLT level at 1 week after treatment in the study group was significantly lower than that in the control group (×10 9 /L: 101±42 vs. 128±59, P < 0.01), there was no significant difference in WBC, LYM or Hb at different time points after treatment between the two groups., Conclusions: Clinical efficacy of lymphocyte apheresis combined with plasma exchange based on comprehensive medical treatment in the treatment of patients with hepatitis B virus-related liver failure at the ascending stage is superior to conservative medical treatment alone, which can improve clinical improvement rate and recovery rate of liver function with high safety.

Artemisiae Argyi Folium, the dried leaves of Artemisia argyi, has been widely used in traditional Chinese and folk medicines for treatment of hemorrhage, pain, and skin itch. Phytochemical studies indicated that volatile oil, organic acid and flavonoids were the main bioactive components in Artemisiae Argyi Folium. Compared to the volatile compounds, the research of nonvolatile compounds in Artemisiae Argyi Folium are limited. In the present study, an accurate and reliable fingerprint approach was developed using HPLC for quality control of Artemisiae Argyi Folium. A total of 10 common peaks were marked，and the similarity of all the Artemisiae Argyi Folium samples was above 0.940. The established fingerprint method could be used for quality control of Artemisiae Argyi Folium. Furthermore, an HPLC method was applied for simultaneous determination of seven bioactive compounds including five organic acids and two flavonoids in Artemisiae Argyi Folium and Artemisiae Lavandulaefoliae Folium samples. Moreover, chemometrics methods such as hierarchical clustering analysis and principal component analysis were performed to compare and discriminate the Artemisiae Argyi Folium and Artemisiae Lavandulaefoliae Folium based on the quantitative data of analytes. The results indicated that simultaneous quantification of multicomponents coupled with chemometrics analysis could be a well-acceptable strategy to identify and evaluate the quality of Artemisiae Argyi Folium., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

To study the correlation between the content of saikosaponins ingredient of Bupleuri Radix and topographic factors, we researched the ecology suitability regionalization of topographic of Bupleuri Radix from Hebei province to provide a scientific basis for selecting artificial planting. Based on 43 samples of Bupleuri Radix from Hebei province, the variation of the content of saikosaponins in different conditions of topographic factors and the influence of slope, altitude and aspect were comprehensively analyzed by SPSS 21.0. Then we studied topographic factors of ecology suitability regionalization of Bupleuri Radix on the basis of the relationship between the saikosaponins and topographic factors by ArcGIS. The most suitable conditions of topographic for cultivation of saikosaponins are as follows：altitude 600 m above, slope 4.00-5.50 degrees, aspect to the sun. In Hebei province, it is suitable for growth of Bupleuri Radix in the Taihang Mountains and the Yanshan Mountains where the content of saikosaponins is higher., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Neural stem cells (NSCs) offer great promise for the treatment of multiple neurodegenerative diseases. However, the survival and differentiation rates of grafted cells in the host brain need to be enhanced. In this regard, understanding of the underlying mechanism of NSCs survival and death is of great importance for the implications of stem cell-based therapeutic application in the treatments of neurological disorders. Autophagy is a conserved proteolytic mechanism required for maintaining cellular homeostasis, which can affect NSCs fate through regulating their biological behaviors, such as survival and proliferation. In this mini-review, we will summarize the effects of autophagy on NSCs fate including survival, apoptosis, proliferation and differentiation, as well as the underlying mechanisms.

ATP-sensitive potassium channels (K ATP ), as an inward rectifying potassium channel, are widely distributed in many types of tissues. K ATP are activated by the depletion of ATP level and the increase in oxidative stress in cells. The activity of K ATP couples cell metabolism with electrical activity and results in membrane hyperpolarization. K ATP are ubiquitously distributed in the brain, including substantia nigra, hippocampus, hypothalamus, cerebral cortex, dorsal nucleus of vagus and glial cells, and participate in neuronal excitability, mitochondria homeostasis and neurotransmitter release. Accumulating lines of evidence suggest that K ATP are the major contributing factors in the pathogenesis of Parkinson's disease (PD). This review discussed the association of K ATP with the pathogenic processes of PD by focusing on the roles of K ATP on the degeneration of dopaminergic neurons, the functions of mitochondria, the firing pattern of dopaminergic neurons in the substantia nigra, the α-synuclein secretion from striatum, and the microglia activation.

Objective: To explore the effects of salidroside (sal) on the expressions of Bcl-2, Bax and caspases-3 proteins in cultured rat subventricular zone (SVZ) neural stem cells (NSCs) exposed to hypoxia injury., Methods: Primarily cultured SVZ NSCs from adult SD rats were incubated with salidroside (120 and 240 µmol/L) for 24 h prior to exposure to hypoxia. The cell viability was assessed with MTT assay, and the cell apoptosis was analyzed using TUNEL staining and flow cytometry. Western blotting was performed to detect the expressions of Bcl-2, Bax and caspase-3 in the cells., Results: Salidroside pretreatment of the cells for 24 h resulted in an obvious resistance to hypoxia-induced cell apoptosis and decrement of cell viability (P<0.05). Salidroside also antagonized the effect of hypoxia exposure in lowering Bcl-2/Bax ratio apoptosis of rat neural stem cells and decreased the expression of caspases-3 protein (P<0.05)., Conclusion: Salidroside can significantly resist hypoxia-induced. The neuroprotective effect of salidroside may be related to the modulation of expressions of apoptosis-related proteins.

To study the attachment, proliferation and differentiation of neural stem cells (NSCs) on surface modified PHBHHx films and to establish the theory of PHBHHx application in NSCs-based brain tissue engineering. PHBHHx film was fabricated by a solution-casting method, and the morphology of the film was observed under scanning electron microscopy(SEM). The films were treated by NaOH or lipase, then the surface hydrophilic property was characterized using water contact angle measurement. NSCs were isolated from the cerebral cortex of rat embryos on embryonic day 14.5, and cultured on surface treated PHBHHx films. The morphology of NSCs attached on the film was visualized under SEM, and the survival and differentiation of NSCs were observed through immunocytochemical staining. Compared with the untreated PHBHHx films, the water contact angle of NaOH or lipase treated PHBHHx films decreased dramatically, and the number of NSCs attached significantly increased. NSCs survived well on treated PHBHHx films and differentiated into neurons and glial cells. The amelioration of hydrophilic property of PHBHHx film improved its biocompatibility with NSCs. PHBHHx can serve as a novel CNS tissue engineering biomaterial applied for NSCs transplantation, brain repairing and regeneration.

Objective: To establish a method for organotypic slice culture of neonatal rat cortex in a modified condition and investigate the effect of spatial signals on neural stem cell (NSC) differentiation., Methods: The brain slices (200 µm in thickness) of neonatal SD rats (3 to 5 days old) were prepared and cultured in modified serum-free DMEM/F12 medium at 37 degrees celsius; with 95% O(2) and 5% CO(2). The organotypic slice cultures were observed regularly. NSCs isolated from the cortex of rat embryos (14-15 embryonic days) were cultured in serum-free DMEM/F12 supplemented with B27 and N2, and the passage 3 NSCs were labeled by CM-DiI before transplanted onto the organotypic slices cultured for 2 weeks. The survival of transplanted NSCs was assessed, and the cell differentiation was identified by immunofluorescence staining., Results: The organotypic slice cultures were well maintained for at least 4 weeks in the modified medium. The thickness of the organotypic slices reduced from 200 µm to 130 µm after 2-week culture in vitro due to the migration of the cells on the edge of the slices. CM-DiI-labeled NSCs survived well and differentiated into GFAP(+) glia and β-tubullin III(+) neurons., Conclusion: Neonatal rat organotypic brain slice can be successfully cultured in a modified condition to serve as a model for studying NSC differentiation induced by spatial signals.