Study on the Expression Levels of Serum miR-139-5p,HDAC4 and GFAP in Neonates with Hypoxic-ischemic Encephalopathy and Their Clinical Value / 现代检验医学杂志

Objective To analyze the relationship between serum micro RNA(miR)139-5p,histone deacetylase 4(HDAC4)and glial fibrillary acidic protein(GFAP)and the severity of brain injury in neonatal hypoxic-ischemic encephalopathy(HIE).Methods From January 2017 to March 2022,72 HIE neonates born in Guangyuan Central Hospital were collected as research objects(study group),while 75 healthy full-term newborns were the control group.The expression levels of miR-139-5p and HDAC4 in serum were detected by real-time fluorescence quantitative PCR.ELISA was applied to detect serum GFAP level.Binary logistic regression was applied to analyze the factors affecting the occurrence of severe brain injury in children with HIE.Results Compared with the control group,the serum GFAP(1.30±0.37ng/L vs 0.50±0.15 ng/L)and HDAC4 relative expression level(2.05±0.39 vs 1.02±0.21)in the study group were increased,the relative expression level of miR-139-5p(0.63±0.14 vs 1.01±0.22)and the NBNA score(33.20±1.43 score vs 39.85±2.23 score)was decreased,the differences were statistically significant(t=17.304,20.046,12.436,21.424,all P＜0.05).Compared with the mild to moderate group,the serum GFAP level(1.61±0.47ng/L vs 1.16±0.33ng/L),HDAC4 relative expression level(2.43±0.37 vs 1.87±0.40),miR-139-5p(0.38±0.10 vs 0.74±0.16)and NBNA score(30.52±1.54 score vs 34.46±1.38 score)relative expression level in the severe group were increased,and the differences were statistically significant(t=4.690,5.669,9.900,10.884,all P＜0.05).Logistic regression analysis showed that low expression of miR-139-5p,high expression of HDAC4,low NBNA score and low Apgar score within 1 min after birth were risk factors for severe brain injury in HIE children(Wald χ2=5.772～6.969,OR=1.519～1.709,all P＜0.05).Pearson analysis showed that the expression level of serum miR-139-5p was negatively correlated with GFAP,HDAC4(r=-0.416,-0.579,all P＜0.05),while the expression level of serum HDAC4 was positively correlated with GFAP(r= 0.437,P＜0.05).Spearman analysis showed that the expression level of serum miR-139-5p was positively correlated with NBNA score,Apgar score within 1 min after birth,and Apgar score within 5 min after birth(r= 0.398,0.367,0.348,all P＜0.05).Serum HDAC4 expression level was negatively correlated with NBNA score,Apgar score within 1 min after birth,and Apgar score within 5 min after birth(r=-0.364,-0.345,-0.332,all P＜0.05).Conclusion The expression of miR-139-5p in the serum of children with HIE was decreased,and the expression of HDAC4 was increased,miR-139-5p and HDAC4 were associated with the severity of brain injury in children with HIE.

Effects of olfactory three needle on apolipoprotein E and pathological substrates in the hippocampus of mice with Parkinson's disease dementia / 中国康复医学杂志

Objective:To explore the mechanism of olfactory three needle therapy on Parkinson's disease dementia(PDD)by observing its effects on expression of apolipoprotein E(ApoE)、glial fibrillary acidic protein(GFAP)and related core pathology substrates in the hippocampus of PDD model mice. Method:Male C57BL/6 mice were randomly divided into four groups:control group(Control),sham opera-tion group(Sham),model group(Model)and acupuncture electrotherapy group(AE),with 10 mice in each group.The PD model was established by injecting 6-OHDA into the medial forebrain tract(MFB)and PDD mice were selected.After successful modeling and selection,the AE group received"olfactory three needle"electro acupuncture treatment.After 14 days of intervention,Morris water maze test and shuttle box test were used to evaluate the learning and memory ability of mice in each group.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of hippocampal CA1 region.Western blotting was used to detect the expression of α-syn,Aβ and ApoE proteins in hippocampal CA1 region.The co-location rate of ApoE and GFAP in hippocampal CA1 region was observed by double immunofluorescence markers. Result:Compared with Model group,the AE group exhibited a shortened escape latency in water maze(P＜0.01),increased platform crossing(P＜0.05),increased active escape times of shuttle box(P＜0.05),and reduced the average total time of electric stimulation(P＜0.01).In the Model group,the neurons in the hippocampal CA1 area were sparsely arranged and showed signs of degeneration and necrosis;and cell nuclei were small,hyperchromatic and had unclear structures,indicating the appearance of nuclear pyrosis.In contrast,the AE group showed significant improvements in neuronal pathology,with most cells regularly arranged,round and large nuclei,lightly stained and clearly shaped.Compared with the Model group,the expression levels of α-syn,Aβ,ApoE protein and the co-localization rate of ApoE and GFAP in hippocampal CA1 region in AE group were decreased(P＜0.01,P＜0.01,P＜0.01,P＜0.05). Conclusion:The"Olfactory three needle"acupuncture can improve the cognitive ability and restore the morpho-logical structure and function of neurons in PDD mice.The mechanism may be related to the inhibition of ApoE expression in astrocytes and the reduction of α-syn and Aβ deposition in hippocampal CA1 region.

Optimization and methods of culture in vitro of astrocytes from cerebral cortical mice / 安徽医科大学学报

Objective @#To explore and optimize the in vitro primary culture method of astrocytes in neonatal mouse cerebral cortex , which provides a better solution for the in vitro culture of astrocytes.@*Methods @#In order to optimize the in vitro culture method of mouse cerebral cortex astrocytes , 3 ⁃day⁃old C57BL/6J mouse cerebral cortex tissues were taken , meninges and blood vessels were removed , digested by pancreatic enzymes and centrifuged , andhigh⁃glucose dulbecco ′s modified eagle medium (DMEM) was added to form cell suspension , which was purified by differential adhesion method , cross hand method and constant temperature shaking method.The cells were inoculated in poly⁃D ⁃lysine⁃coated culture bottles with different culture densities , and the purity of astrocytes was determined by morphological ob servation and immunofluorescence staining.@*Results @#The cells were inoculated at a density of 5 × 106 cells per bottle with good effect and high activity. The purity of astrocytes reached 99% by using high sugar DMEM medium combined with differential adhesion method , cross hand method and constant temperature shaking method.@*Conclusion @#The primary culture method of astrocytes in mouse cerebral cortex is successfully established and optimized.

Effect of Anmeidan on Serum Levels of BDNF， GFAP， and Irisin in Patients with Chronic Insomnia / 中国实验方剂学杂志

ObjectiveTo explore the effect of Anmeidan on the sleep quality and serum levels of brain-derived neurotrophic factor （BDNF）， glial fibrillary acidic protein （GFAP）， and irisin in the patients with chronic insomnia. MethodA multicenter， randomized， double-blind， placebo-controlled clinical study was carried out， including 480 patients with chronic insomnia （deficiency syndrome） in Wuhan （Hubei）， Guangzhou （Guangdong）， and Lanzhou （Gansu）. They were randomized into an observation group and a control group at a ratio of 1∶1. The observation group was orally administered with Anmeidan granules at a dose of 11 g， 3 times per day， and the control group with Anmeidan simulant at a dose of 11 g， 3 times per day， Both groups of patients received sleep education after enrollment. After 4 weeks of medication， the Athens insomnia scale （AIS） scores， Spiegel scale scores， and serum levels of BDNF， GFAP， and irisin were compared between the two groups as well as between before and after treatment. ResultA total of 480 adult patients with chronic insomnia were enrolled in this study， with 64 patients falled off. Finally， the 415 patients were included in the analysis， including 213 patients in the observation group and 202 patients in the control group. There was no difference in age or sex between the two groups of patients. Compared with before treatment， the treatment in both groups decreased the AIS and Spiegel scores （P<0.01）. After treatment， the observation group had lower AIS and Spiegel scores than the control group （P<0.01）. The treatment in the observation group slightly lowered the level of BDNF， elevated the level of irisin （P<0.05）， and lowered the level of GFAP （P<0.05） in the serum. After treatment， the observation group showed higher level of irisin （P<0.05） and lower levels of BDNF and GFAP in the serum than the control group. ConclusionAnmeidan may improve the sleep quality of patients with chronic insomnia by elevating the irisin level and lowering the GFAP level in the serum.

Clinical features of autoimmune glial fibrillary acidic protein astrocytopathy in children: an analysis of 34 cases / 中国当代儿科杂志

OBJECTIVES@#To study the clinical features of children with autoimmune glial fibrillary acidic protein astrocytopathy (GFAP-A).@*METHODS@#A retrospective analysis was performed on the medical data of 34 children with GFAP-A who attended the Department of Neurology, Children's Hospital of Chongqing Medical University, from January 2020 to February 2022. The medical data included clinical manifestations, cerebrospinal fluid features, imaging examination results, treatment, and prognosis.@*RESULTS@#The median age of onset was 8.4 (range 1.9-14.9) years for the 34 children with GFAP-A. The main clinical manifestations included headache (50%, 17/34), fever (47%, 16/34), visual impairment (47%, 16/34), and disturbance of consciousness (44%, 15/34). Abnormal cerebrospinal fluid results were observed in 19 children (56%, 19/34), among whom 8 children had positive autoantibody. The children with overlap syndrome had significantly higher recurrence rate and rate of use of immunosuppressant than those without overlap syndrome (P<0.05). About 77% (24/31) of the children had good response to immunotherapy, and only 1 child had poor prognosis.@*CONCLUSIONS@#Children with GFAP-A often have non-specific clinical symptoms and show good response to immunotherapy. Children with overlap syndrome have a high recurrence rate, and early application of immunosuppressants may help to prevent recurrence and alleviate symptoms.

Effects of specific knockdown of ribosomal protein S5 in hepatic stellate cells on liver fibrosis / 药学实践与服务

Objective To observe the effect of specific knockdown of hepatic stellate cells (HSC) ribosomal protein S5 (RPS5) on liver fibrosis in rats. Methods The glial fibrillary acidic protein (GFAP) promoter-driven RPS5 shRNA adenovirus was established, and AdGFa2-shRPS5 and its control AdGFa2 shNC were used to transfect primary rat HSCs and hepatocytes, respectively. RPS5 was determined by Western-blot and Real Time PCR, α-SMA and type I collagen expression; the rat liver fibrosis model was established by dimethyl nitrosamine (DMN) and bile duct ligation (BDL), and intrahepatic HSC was specifically knocked down by tail vein injection of adenovirus of RPS5 levels. The pathological changes of liver tissue sections were analyzed by HE staining; the content of hydroxyproline, sections of Sirius red and Masson staining were used to evaluate collagen deposition; immunohistochemical staining was used to detect the expression of α-SMA and RPS5. Results AdGFa2-shRPS5 specifically knocked down the expression level of RPS5 in HSC and increased the expression of α-SMA and type I collagen in vitro. The in vivo results showed that in two animal models of chronic liver injury, specific knockdown of RPS5 expression in HSCs promoted HSC activation, increased the deposition of extracellular matrix, and promoted liver fibrosis. Conclusion RPS5 is essential for HSC activation and liver fibrosis, which could be a potential target for the treatment of liver fibrosis.

Observation of 4 patients with positive anti-glial fibrillary acidic protein antibody / 中华眼底病杂志

Objective:To observe the clinical characteristics of the patients with positive anti-glial fibrillary acidic protein (GFAP) antibody.Methods:A retrospective study. From January 2017 through December 2021, 4 patients with positive anti-GFAP antibodies hospitalized in Departments of Ophthalmology and Neurology of Xijing Hospital, Air Force Medical University were included in this study. There were 3 patients with optic neuritis (ON) and 1 patient with the spinal and cerebral lesions. All patients were female, with an average age of 35 years. Three patients with ON received the examinations of best corrected visual acuity (BCVA), optical coherence tomography, visual evoked potential and magnetic resonance imaging (MRI) for the head and orbital. Another 1 patient with the spinal and cerebral lesions underwent MRI for the head, cervical and thoracic vertebras. All patients were tested for demyelinating ON-related antibodies in the serum, and the patient with the spinal and cerebral lesions for the antibodies in both serum and cerebrospinal fluid. Patients with ON received intravenous infusion of methylprednisolone sodium succinate in the acute stage, while the patients with spinal cord and brain lesions were given glucocorticoid and immunosuppressive therapy.Results:The initial symptoms of the patients with ON were sudden blurred vision in the right eye together with a pain when the eye rotated. BCVA were hand moving/in-front, 0.2 and 0.12, respectively. The serum anti-GFAP antibodies were positive. MRI showed a rough and thickened optic nerve in 1 patient. For patients with BCVA of hand moving/in-front, the BCVA was increased to counting fingers/30 cm on discharge; while the other 2 patients had no changes for BCVA. When followed up on phone 2-3 years after discharge, BCVA of the patients with ON increased to higher than 0.6. No ocular symptoms occurred in the patient with spinal and cerebral lesions and his initial symptoms were numbness, weakness and convulsions of limbs, accompanied by slurred speech. His anti-GFAP antibodies in the serum were negative but positive in the cerebrospinal fluid. MRI showed enhanced cerebellum and spinal dura mater. The initial symptoms were relieved on discharge, and vanished when followed up on phone after discharge.Conclusions:The patients with positive anti-GFAP antibodies are more common in young and middle-aged women. Monocular optic neuritis is more often seen in the form of sudden blurred vision with an eye-rotating pain. Anti-GFAP antibodies in the serum are positive, and a few patient show a rough and thickened optic nerve. They are sensitive to glucocorticoid therapy with a satisfied prognosis.

Autoimmune glial fibrillary acidic protein astrocytopathy / 中华神经科杂志

Autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy is a rare immune-mediated inflammatory disease of central nervous system reported in recent years, and its specific biological marker is anti-GFAP autoantibody. In this paper, the etiology, pathogenesis, clinical manifestations, auxiliary examination and treatment of the disease are comprehensively expounded, so as to improve the understanding of clinicians, especially neurologists.

Effects of volatile oil from Acori graminei Rhizoma on glial fibrillary acidic protein, c-Jun N-terminal protein kinase and tumor necrosis factor-α expressions in the spinal cord dorsal horn of the inflammatory pain rats / 解剖学报

Objective To investigate the influence of volatile oil from Acori graminei Rhizoma (VOA) on expressions of glial fibrillary acidic protein (GFAP), c-Jun N-terminal protein kainse (JNK) and tumour necrosis factor-α (TNF-α) in the spinal cord dorsal horn of imflammatory pain rats. Methods Totally 36 male SD rats were randomly divided into control group (control), sham-operated group (sham), complete Freund' s adjuvant group (CFA), 5 g/(kg·d) low dose VOA+CFA group (VOA-L+CFA), 10 g/(kg·d) medium dose VOA + CFA group (VOA-M+CFA) and 20 g/(kg·d) high dose VOA + CFA group (VOA-H+CFA). All animals were sacrificed immediately after continuous gavage administration for 22 days. The expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats in each group were detected by immunofluorescence and Western blotting methods. Results The present results showed that the positive expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats increased significantly in the CFA group, when compared to the control and sham groups (P < 0. 01). The expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats with VOA treatment reduced in the dose-dependent manner, when compared to the CFA group, the positive expressions of GFAP, JNK and TNF-α reduced significantly in the dorsal horn of the spinal cord of the VOA-H+CFA group (P<0. 05, P<0. 01). Conclusion VOA reduces the expressions of GFAP, JNK and TNF-α in the spinal cord dorsal horn of rats of CFA-induced inflammatory pain.

Effect of Xiaoxuming Decoction on activation of astrocytes in acute cerebral ischemia/reperfusion injury / 中国中药杂志

This study investigated the effect of Xiaoxuming Decoction(XXMD) on the activation of astrocytes after cerebral ischemia/reperfusion(I/R) injury. The model of cerebral IR injury was established using the middle cerebral artery occlusion method. Fluorocitrate(FC), an inhibitor of astrocyte activation, was applied to inhibit astrocyte activation. Rats were randomly divided into a sham group, a model group, a XXMD group, a XXMD+FC group, and a XXMD+Vehicle group. Neurobehavioral changes at 24 hours after cerebral IR injury, cerebral infarction, histopathological changes observed through HE staining, submicroscopic structure of astrocytes observed through transmission electron microscopy, fluorescence intensity of glial fibrillary acidic protein(GFAP) and thrombospondin 1(TSP1) measured through immunofluorescence, and expression of GFAP and TSP1 in brain tissue measured through Western blot were evaluated in rats from each group. The experimental results showed that neurobehavioral scores and cerebral infarct area significantly increased in the model group. The XXMD group, the XXMD+FC group, and the XXMD+Vehicle group all alleviated neurobehavioral changes in rats. The pathological changes in the brain were evident in the model group, while the XXMD group, the XXMD+FC group, and the XXMD+Vehicle group exhibited milder cerebral IR injury in rats. The submicroscopic structure of astrocytes in the model group showed significant swelling, whereas the XXMD group, the XXMD+FC group, and XXMD+Vehicle group protected the submicroscopic structure of astrocytes. The fluorescence intensity and protein expression of GFAP and TSP1 increased in the model group compared with those in the sham group. However, the XXMD group, the XXMD+FC group, and XXMD+Vehicle group all down-regulated the expression of GFAP and TSP1. The combination of XXMD and FC showed a more pronounced effect. These results indicate that XXMD can improve cerebral IR injury, possibly by inhibiting astrocyte activation and down-regulating the expression of GFAP and TSP1.

Correlation between plasma glial fibrillary acidic protein, α -synuclein and progression of Parkinson disease / 中华行为医学与脑科学杂志

Objective:To investigate the correlation between plasma glial fibrillary acidic protein (GFAP), total α-synuclein (α-syn), phosphorylated α-synuclein (p-α-syn) and the disease progression in patients with Parkinson disease(PD).Methods:Sixty-three patients with PD including the outpatients and inpatients from Department of Neurology of Huai'an First People's Hospital were continuously collected as a PD group, and 25 healthy people matched in age and sex were selected as a healthy control (HC) group at the same time. The plasma levels of GFAP, α-syn, and p-α-syn levels in the PD group and HC group were detected by ELISA. SPSS 25.0 software was used for statistical analysis. Mann-Whitney U test was performed to assess the differences of GFAP, α-syn and p-α-syn levels between the two groups, and the correlation between GFAP and α-syn and p-α-syn levels in PD group was examined by Spearman correlation analysis. The levels of GFAP, α-syn and p-α-syn in different Hoehn-Yahr(H-Y) stages of PD group were compared by Mann-Whitney U test and Spearman correlation analysis. Results:The levels of GFAP (0.80(0.62, 0.97)ng/mL, 0.54(0.27, 0.88)ng/mL, Z=-3.216, P=0.001), α-syn (3.93(3.16, 6.02)ng/mL, 2.67(1.74, 4.47)ng/mL, Z=-2.600, P=0.009), and p-α-syn (5.80(1.31, 15.62), 0.71(0.61, 0.83), Z=-6.607, P<0.001) in PD group were higher than those in HC group, and the difference was statistically significant. In PD group, there was a significant positive correlation between GFAP and α-syn ( r=0.442, P<0.001) and p-α-syn ( r=0.493, P<0.001). GFAP in the advanced PD group was higher than that in the early PD group, and the difference was statistically significant( P=0.039). There was no significant difference in α-syn and p-α-syn between different H-Y stages of PD patients( P>0.05). Plasma GFAP was positively correlated with H-Y stages ( r=0.277, P=0.018). Conclusion:The level of plasma GFAP in PD patients is positively correlated with disease progression, which can be used as a potential biomarker for PD disease assessment.

Neuroprotective effect of Chuanjin jieyu decoction on chronic stress-induced depression rat model / 中国医师杂志

Objective:To study the neuroprotective effect of Chuanjin Jieyu decoction on a chronic unpredictable stress (CUMS) induced depression rat model.Methods:A total of 48 adult male healthy Sprague-Dawley (SD) rats were randomly selected, with 12 rats as the control group, and the remaining rats were used to establish a CUMS depression model. After successful modeling, the rats were randomly divided into three groups: CUMS group, fluoxetine group, and Chuanjin Jieyu decoction group; 0.9% sodium chloride solution fluoxetine and Chuanjin Jieyu decoction were respectively administered for intervention for 4 weeks. After modeling and intervention, depression levels in rats were measured through open field experiments and sugar water preference experiments; After intervention, real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of brain-derived neurotrophic factor (BDNF), tyrosine protein kinase receptor B (TrkB), and cyclic adenosine monophosphate responsive element binding protein (CREB) mRNA in the dentate gyrus of the hippocampus. Immunofluorescence was used to detect the expression levels of BDNF and NeuN. Western blot was used to detect the level of glial fibrillary acidic protein (GFAP).Results:After modeling, the sugar consumption, sugar preference, vertical and horizontal movement results of the CUMS group, fluoxetine group, and Chuanjin Jieyu decoction group were significantly lower than those of the control group (all P<0.01); After intervention, the above indicators in the CUMS group were lower than those in the control group (all P<0.01). The above indicators in the fluoxetine group and the Chuanjin Jieyu decoction group were higher than those in the CUMS group ( P<0.01 or P<0.05), but lower than those in the control group ( P<0.01 or P<0.05). The qRT-PCR results showed that the mRNA expression levels of BDNF, TrkB, and CREB in the CUMS group, the fluoxetine group, and the Chuanjin Jieyu decoction group were significantly lower than those in the control group (all P<0.01); The above indicators in the fluoxetine group and the Chuanjin Jieyu decoction group were significantly higher than those in the CUMS group (all P<0.01). The immunofluorescence results showed that the expression levels of BDNF and NeuN in the CUMS group were significantly lower than those in the control group (all P<0.01); The above indicators in the fluoxetine group and the Chuanjin Jieyu decoction group were higher than those in the CUMS group ( P<0.01 or P<0.05), and all indicators in the Chuanjin Jieyu decoction group and the NeuN expression level in the fluoxetine group were lower than those in the control group ( P<0.01 or P<0.05). Western blot results showed that the expression of GFAP protein in the CUMS group was lower than that in the control group ( P<0.01); The expression of GFAP protein in the fluoxetine group was higher than that in the CUMS group ( P<0.05). Conclusions:Chuanjin Jieyu decoction can improve the behavioral indicators of depression in rats, increase the expression of BDNF, TrkB, CREB, and NeuN in the hippocampal dentate gyrus, suggesting that Chuanjin Jieyu decoction may exert neuroprotective effects through the BDNF/TrkB/CREB pathway.

Progress of major biomarkers for neonatal brain injury in clinical application / 国际儿科学杂志

With the continuous progress of monitoring and treatment skills, the mortality of neonates has gradually decreased, and the long-term neurodevelopmental outcome has become the primary concern of society and families.During the perinatal period, the developing brain is vulnerable to hypoxia, hemorrhage, infection and inflammation, which may cause varying degrees of brain cell damage.Studies have found that proteins released by damaged brain cells can be detected in the body fluid of neonates, which are related to the occurrence and prognosis of neonatal brain injury.This article mainly reviews the recently reported brain injury biomarkers such as S100B, neuron specific enolase(NSE)and glial fibrillary acidic protein(GFAP)in different biological samples and its clinical predictive value for the occurrence of brain injury and neurodevelopmental prognosis.

Effect of providing purple sweet potato water extract on tumor necrosis factor-α levels, protein 53 expression, glial fibrillary acidic protein expression, brain-derived neurotrophic factor levels, and spatial working memory in rats with d-galactose induction / Efeito do extrato de água de batata doce roxo no fator de necrose tumoral níveis alfa, na expressão da proteína 53, na expressão de proteína ácida fibrilar glial, nos níveis de fator neurotrófico cerebral e na memória espacial em ratos com indução de d-galactose

ABSTRACT. Alzheimer's dementia (AD) is a neurodegenerative disease. The mechanism of oxidative stress in AD is due to amyloid beta (Aβ) protein that aggregates to form plaques, which further triggers chronic inflammation and neuronal apoptosis. Purple sweet potato extract with the main content of anthocyanins is a potential antioxidant with a direct target on the amyloid cascade hypothesis. Objective: The research objective was to determine the role of purple sweet potato water extract as an antioxidant and anti-inflammatory in preventing apoptosis in order to provide a neuroprotective effect in d-galactose-induced rats. Methods: A total of 100 male Wistar rats with randomized posttest-only control group design that met the eligibility criteria were included in this study. The treatment group was given 200 mg/kg BW/day of purple sweet potato water extract on days 1-70. d-galactose induction was administered in the treatment and control groups on days 15-70. Results: The independent t-test showed that the mean tumor necrosis factor-α (TNF-α) levels in the treatment group (735.36±139.74) was significantly lower than that in the control group (896.77±152.52). The p53 and glial fibrillary acidic protein (GFAP) expressions of astrocyte cells in the treatment group were significantly lower than that in the control group. The brain-derived neurotrophic factor (BDNF) levels in the treatment group (498.13±121.47) were higher than that in the control (391.93±140.28), and there was a significant increase in spatial working memory in the treatment group (72.01±10.22) than the control (59.77±11.87). Conclusions: The neuroprotective effect of purple sweet potato extract is due to d-galactose induction resulting from decrease in TNF-α levels, p53 expression, and GFAP expression and increase in BDNF levels and spatial working memory.

RESUMO. A doença de Alzheimer (DA) é uma doença neurodegenerativa. O mecanismo de estresse oxidativo na DA ocorre devido à proteína beta amilóide que se agrega para formar placas que desencadeiam inflamação crônica e apoptose neuronal. O extrato de batata-doce roxa composto principalmente por antocianinas é um potencial antioxidante com efeito direto sobre a hipótese da cascata amilóide. Objetivo: O objetivo da pesquisa foi determinar o papel do extrato aquoso de batata-doce roxa como antioxidante e anti-inflamatório na prevenção da apoptose, para proporcionar um efeito neuroprotetor em ratos induzidos por D-galactose. Métodos: Grupo controle randomizado pós-teste com 100 ratos Wistar machos que preencheram os critérios de elegibilidade. O grupo de tratamento recebeu 200mg/kg de peso corporal/dia de extrato aquoso de batata-doce roxa nos dias 1-70. A indução de D-galactose foi testada nos grupos de tratamento e controle nos dias 15-70. Resultados: O teste t independente mostrou que a média dos níveis de TNF-α no grupo de tratamento (735,36±139,74) foi significativamente menor do que no grupo controle (896,77±152,52). A expressão de p53 e a expressão de GFAP de células de astrócitos foram significativamente menores no grupo de tratamento do que no grupo controle. Os níveis de BDNF no grupo de tratamento (498,13±121,47) foram maiores que no grupo controle (391,93±140,28) e houve um aumento significativo da memória de trabalho espacial no grupo de tratamento (72,01±10,22) em relação ao controle (59,77±11,87). Conclusões: O efeito neuroprotetor do extrato de batata-doce roxa é devido à indução de D-galactose pela diminuição dos níveis de TNF-α, expressão de p53 e expressão de GFAP, aumentando assim os níveis de BDNF e memória espacial.

Clinical significance of prognostic serum marker expression in older adult patients with sepsis-associated encephalopathy / 中国基层医药

Objective:To investigate the clinical significance of prognostic serum marker expression in older adult patients with sepsis-associated encephalopathy (SAE).Methods:The clinical data of 79 older adult patients with SAE who received treatment in The Second People's Hospital of Hefei from June 2019 to February 2021 (study group) and 121 sepsis patients without encephalopathy concurrently (control group) were retrospectively analyzed. The indexes with statistically significant difference between the two groups were subjected to multivariate binary logistic regression. Survival curve was plotted.Results:There were no significant differences in neuron specific enolase [NSE, (10.69 ± 4.31) μg/L vs. (24.84 ± 3.28) μg/L, t = 26.25, P < 0.01], S100β [(0.25 ± 0.06) μg/L vs. (0.53 ± 0.09) μg/L, t = 22.45, P < 0.01], monocyte chemoattractant protein-1 [MCP-1, (99.33 ± 4.87) ng/L vs. (179.99 ± 6.02) ng/L, t = 99.94, P < 0.01], malondialdehyde [MDA, (4.22 ± 0.08) nmol/L vs. (6.78 ± 0.11) nmol/L, t = 33.76, P < 0.01], glial fibrillary acidic protein [GFAP, (0.21±0.08) μg/L vs. (2.03 ± 0.47) μg/L, t = 33.76, P < 0.01], procalcitonin [(7.04 ± 2.50) ng/L vs. (16.23 ± 2.48) ng/L, t = 25.47, P < 0.01], interleukin-6 [(29.91 ± 4.51) ng/L vs. (69.22 ± 6.79) ng/L, t = 45.51, P < 0.01], Acute Physiology and Chronic Health Evaluation II (APACHE II) score [(18.33 ± 2.12) points vs. (28.89 ± 5.09) points, t = 17.53, P < 0.01], and sequential organ failure assessment score [(7.69 ± 1.50) points vs. (14.05 ± 1.55) points, t = 28.92, P < 0.01] between the control and study groups. N-terminal pro B-type natriuretic peptide was (868.38 ± 25.28) ng/L and (1 037.19 ± 25.34) ng/L in the control and study groups, respectively. Logistic regression analysis revealed that NSE, MCP-1, MDA, and GFAP were the independent risk factors for developing SAE in older adults (NSE: t = 8.42, P < 0.01; MCP-1: t = 4.16, P < 0.01; MDA: t = 18.4, P < 0.01; GFAP: t = 2.88, P < 0.01). The survival curve indicated that survival rate was significantly lower in the study group than in the control group. Conclusion:NSE, MCP-1, MDA, and GFAP are independent risk factors for developing SAE in older adults.

Effect of vagus nerve stimulation by electroacupuncture on GFAP and NeuN in hippocampal of juvenile rats with depression induced by chronic unpredictable stress / 中国医师杂志

Objective:To analyze the expression of glial fibrillary acidic protein (GFAP) and neuronal nuclei (NeuN) antigen in hippocampus based on the depression model of juvenile rats caused by chronic unpredictable stress (CUS), and to explore the effect of electroacupuncture vagus nerve on CUS depression model.Methods:Six juvenile SD rats were selected as the control group (without any stimulation), and the rest were divided into CUS group, pseudo stimulation group, fluoxetine group and electroacupuncture group by random number method after CUS modeling, with 6 rats in each group. Fluoxetine group was given 10 mg/kg fluoxetine intervention; control group and CUS group were given the same amount of normal saline intervention; In the electroacupuncture group, the distal vagus nerve was stimulated after ligation, while in the pseudo stimulation group, only vagus nerve was isolated without electrical stimulation. After 28 d of intervention, the five groups were subjected to Open-field Test and Sucrose Preference Test. Hippocampal neurons were detected by hematoxylin and eosin (HE) staining, and the expressions of GFAP and NeuN in hippocampal were detected by immunohistochemistry.Results:After CUS modeling and before intervention, the number of vertical and horizontal movements, sucrose consumption and sucrose preference in CUS group, pseudo stimulation group, fluoxetine group and electroacupuncture group were significantly lower than those in the control group (all P<0.01); After the intervention, the above indexes in CUS group and pseudo stimulation group were still lower than those in the control group (all P<0.01), but the above indexes in fluoxetine group and electroacupuncture group were significantly higher than those in CUS group and pseudo stimulation group (all P<0.01). HE staining showed that the arrangement of hippocampal neurons in CUS group and pseudo stimulation group were loose, and there were cell swelling and pyknosis, which was significantly improved in fluoxetine group and electroacupuncture group. Immunohistochemical results showed that compared with the control group, the expression of GFAP increased and NeuN decreased in the hippocampus of CUS group and pseudo stimulation group (all P<0.01); Compared with CUS group and pseudo stimulation group, the expression of GFAP decreased and NeuN increased in fluoxetine group and electroacupuncture group (all P<0.01). Conclusions:Electroacupuncture of vagus nerve can obviously improve the depression symptoms of juvenile rats, which is similar to fluoxetine, and may be related to regulating the expression of GFAP and Neun in hippocampus.

Protective Effects of Anmeidan on Cell Structure Against Neuronal Damage in Hippocampal CA1 Region of Sleep-deprived Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effects of Anmeidan （AMD） on neuronal structure and neuronal marker protein expression in the hippocampal CA1 region of sleep-deprived （SD） rats. MethodRats were randomly divided into control group， model group， an AMD group （9.09 g·kg-1·d-1）， and melatonin group （0.27 g·kg-1·d-1）. Rats in the control group and the model group received equal volumes of physiologicol saline. The SD model was induced by the self-made sleep deprivation box for four weeks. Ethovision XT system detected and analyzed the spontaneous behaviors of rats. The histomorphology of neurons in the hippocampal CA1 region was observed by hematoxylin-eosin （HE） staining and Nissl staining， and the changes in Nissl bodies were observed by Nissl staining. The ultrastructure of hippocampal cells was observed by transmission electron microscopy （TEM）. Immunohistochemistry was used to detect the expression of glial fibrillary acidic protein （GFAP）， microtubule-associated protein 2 （MAP2）， nestin， and neuronal nuclei （NeuN） in the CA1 region. ResultCompared with the control group， the model group showed longer distance， increased average activity speed， cumulative duration， average body fill， and higher activity frequency （P<0.01）. Besides， the neurons in the CA1 region were reduced in number with disorganized arrangement， wrinkled nuclei， deeply stained cytoplasm， reduced Nissl bodies， swollen and deformed mitochondria， shortened cristae， and swollen Golgi vesicles. Furthermore， the mean integral absorbance （IA） value of GFAP increased and those of MAP2， nestin， and NeuN decreased （P<0.01）. Compared with the model group， the AMD group showed shortened distance traveled， lower average activity speed， shorter cumulative duration， decreased average body fill， and reduced activity frequency （P<0.05， P<0.01）. Moreover， the neurons in the CA1 region were relieved from damage with increased cell number， clear nuclei and cytoplasm， increased Nissl bodies， and relieved mitochondrial damage. The IA value of GFAP decreased and those of MAP2， nestin， and NeuN increased （P<0.05， P<0.01）. ConclusionAMD can improve structural damage of neurons in the hippocampal CA1 region of sleep-deprived rats， which may be achieved by decreasing GFAP expression and increasing MAP2， nestin， and NeuN expression.

" Multi-hits" based SD rat models of cognitive dysfunction and research on related mechanisms / 中华实用儿科临床杂志

Objective:To establish Sprague-Dawley (SD) rat models of cognitive impairment through repeated stimulation of lipopolysaccharide (LPS) in the early brain development, and to inquire into the effect of " multi-hits" mediated by inflammatory response on the histology and behavior of SD rat models and related molecular mechanisms.Methods:This study adopted a group design for experiments.The " multi-hits" SD rat models were established by intraperitoneal injection of LPS.According to the random number table method, 24 pregnant rats were randomly divided into 4 groups: control group, LPS1 group, LPS2 group and LPS3 group, 6 rats in each group.In the control group, saline was intraperitoneally injected into rats with gestational age of 18 days and 20-day-old neonatal rats.Rats with gestational age of 18 days were intraperitoneally injected with saline in the LPS1 group, 0.05 mg/kg LPS in the LPS2 group, and 0.1 mg/kg LPS in the LPS3 group.The pups in LPS1-3 groups were all injected intraperitoneally with 1 mg/kg LPS at the postnatal age of 20 days.The motor and cognitive function of the pups were evaluated overall by behavioral experiments such as forelimb suspension tests, grid tests and water maze tests.The relative expression of glial fibrillary acidic protein (GFAP), Notch1 and Jagged1 in brain tissue of pups was mainly detected by Western blot (WB) and histological experiments.One-way ANOVA analysis of variance and independent samples t- test were used to compare data among groups and between groups, respectively. Results:(1) Behavioral experiments: compared with the control group, LPS1-3 groups showed progressive decrease in forelimb suspension time [(34.81±5.66) s, (22.47±4.35) s, and (13.20±4.25) s vs.(43.88 ± 4.85) s], and the number of missteps in the grid experiment increased progressively (16.13±2.90, 20.75±3.10, 25.13±4.45 vs.9.00±2.72). The differences were statistically significant ( F=69.77, 35.59, all P<0.001). Both the escape latency and total distance in Morri′s water maze test increased progressively ( P<0.05). (2) WB experiment: the relative expression levels of GFAP, Notch1 and Jagged1 proteins in LPS1-3 groups were significantly higher than that in the control group ( P<0.05). (3) Hematoxylin-eosin (HE) staining and electron microscope pathology: compared with the control group, LPS1-3 groups had more loosely arranged frontal cortices and more obvious cell pyknosis.Under the electron microscope, the cytoplasm was swelling to varying degrees, mitochondrial cristae were broken, and part of the nuclear membrane was damaged. Conclusions:In the " multi-hits" cognitive impairment model, the damage to the brain tissue structure and behavioral changes of pups may be related to the up-regulation of Notch1/Jagged1 pathway mediated by repeated exposure to LPS.

Effects of excessive fluoride exposure on astrocytes and the expression of glial fibrillary acidic protein in vitro and in vivo / 中华地方病学杂志

Objective:To investigate the effects of excessive fluoride exposure on astrocytes and the expression of glial fibrillary acidic protein (GFAP), in vitro and in vivo. Methods:(1) In vivo experiment: 24 SPF SD rats, half male and half female, were randomly divided into control and fluoride exposed groups according to sex and body weight, 12 rats in each group. Rats were fed with < 1 mg/L and 50 mg/L sodium fluoride solution prepared by tap water for 6 months, respectively. The expression levels of GFAP protein in rat brain tissue were measured by immunofluorescence, immunohistochemistry and Western blotting. (2) In vitro experiment: adult (6-month-old) rat cortical astrocytes were extracted and cultured in primary culture (4 mmol/L sodium fluoride solution for 24 h), and the astrocytes were identified by immunofluorescence, and GFAP mRNA and protein expression levels were detected by real-time fluorescence quantitative PCR and Western blotting, respectively, and astrocytes apoptosis and calcium ion content were detected by flow cytometry. Results:(1) In vivo experiment: the results of immunofluorescence, immunohistochemistry and Western blotting showed that the GFAP protein expression level in brain tissue of rats exposed to fluoride was higher than that of control group (0.440 ± 0.200 vs 0.250 ± 0.120, t =-5.93, P = 0.027; 0.270 ± 0.020 vs 0.240 ± 0.050, t =-4.87, P = 0.040; 1.017 ± 0.001 vs 0.486 ± 0.006, t =-52.48, P = 0.001). (2) In vitro experiment: GFAP positive cells were identified as astrocytes by immunofluorescence; GFAP mRNA expression level was higher in fluoride exposed group than that of control group by real-time fluorescence quantitative PCR (2.780 ± 0.120 vs 0.134 ± 0.005, t =-37.84, P = 0.001). The Western blotting results showed that the GFAP protein expression level was higher in fluoride exposed group than that of control group (2.76 ± 0.10 vs 1.38 ± 0.05, t =-20.44, P = 0.002). Flow cytometry results showed that the apoptosis rate of astrocytes was higher in fluoride exposed group than that of control group (%: 55.0 ± 1.0 vs 3.5 ± 0.6, t =-10.28, P = 0.009) and the calcium ion content was lower than that of control group (%: 54 ± 9 vs 72 ± 13, t = 4.64, P = 0.043). Conclusion:Excessive fluoride exposure causes increased GFAP expression in astrocytes in vitro and in vivo, promotes apoptosis, and affects calcium signaling pathways.

Clinical Features of Multicenter Autoimmune Glial Fibrillary Acidic Protein Astrocytosis / 罕见病研究

  Objective  To investigate the clinical features of autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy.  Methods  We collected and analyzed the clinical and laboratory data and obtained the clinical characteristics of diagnosis and treatment from fifteen patients with positive GFAP antibody tested by cerebrospinal fluid and diagnosed autoimmune GFAP astrocytopathy by the multi-centers.  Results  The mean age of the first onset of autoimmune GFAP astrocytopathy was 39.73 years old (range 4-65 years), with no significant gender difference. In terms of clinical manifestations, we found the whole brain symptoms including abnormal mental behavior, disturbance of consciousness, epileptic attack accounting for more than 50, , meningitis accounting for 66.7%, myelitis (53.3%), limb tremor (53.3%), vision loss (33.3%); systemic symptoms including fever(100%) and fatigue(86.7%). 46.7% of patients were initially diagnosed with tuberculous meningoencephalitis and were treated with diagnostic antituberculous therapy. The MRI showed 46.7% of patients showed brain linear perivascular radial gadolinium enhancement in the white matter perpendicular to the ventricle.  Conclusions  Autoimmune GFAP astrocytopathy are acute or subacute dieases and the main clinical features include encephalitis, meningitis, myelitis and optic neuritis. They are likely to be misdiagnosed as tuberculous meningoencephalitis and can manifest progressive loss of consciousness in early phase, which is even life threatening.