Journal: Experimental and Therapeutic Medicine

Article Title: Involvement of neurotrophic signaling in doxorubicin-induced cardiotoxicity

doi: 10.3892/etm.2019.8276

Figure Lengend Snippet: Effects of DOX on the expression of BDNF and NGF in the serum and heart. (A) BDNF concentration in the serum. (B) BDNF mRNA levels in the heart. (C) BDNF protein expression levels in the heart. (D) NGF concentration in the serum. (E) NGF mRNA levels in the heart. (F) NGF protein levels in the heart. Data are expressed as the mean ± standard deviation (n=8–9). *P<0.05 and **P<0.01 compared with Con. NGF, nerve growth factor; BDNF, brain derived neurotrophic factor; DOX, doxorubicin; Con, control.

Article Snippet: Determination of serum BDNF and NGF levels ELISA kits were purchased from Wuhan USCN Business Co., Ltd. and were used to determine plasma levels of BDNF (cat. no. SEA011Ra) and NGF (cat.no.

Techniques: Expressing, Concentration Assay, Standard Deviation, Derivative Assay

Journal: PLoS ONE

Article Title: Peripheral blood levels of brain-derived neurotrophic factor in patients with post-traumatic stress disorder (PTSD): A systematic review and meta-analysis

doi: 10.1371/journal.pone.0241928

Figure Lengend Snippet: A meta-analysis of BDNF levels in PTSD patients compared to non-PTSD controls by the technique used for BDNF measurement (ELISA and sandwich ELISA).

Article Snippet: Chemicon, Promega, Mallipore ChemiKine and Milliplex, Picokine, Raybiotech, Human BDNF ELISA, and Nanjing Jiancheng ELISA kits for human BDNF were the other used kits in the studies.

Techniques: Enzyme-linked Immunosorbent Assay, Sandwich ELISA

Journal: Translational Psychiatry

Article Title: Brain-derived neurotrophic factor protects against tau-related neurodegeneration of Alzheimer's disease

doi: 10.1038/tp.2016.186

Figure Lengend Snippet: Brain-derived neurotrophic factor (BDNF) protein levels in serum and brain are reduced in Alzheimer's disease (AD) patients and P301L transgenic mice. ( a) Comparison of BDNF levels in serum between AD patients and age- and sex-matched healthy elderly controls (HE; mean±s.e.m., Student's t -test, ** P <0.01). ( b ) Detection rate of BDNF in cerebrospinal fluid (CSF) by enzyme-linked immunosorbent assay (ELISA; %, χ 2 -test, * P <0.05). ( c ) Comparison of BDNF levels in serum between AD patients and HE controls (detected by ELISA) (mean±s.e.m., Student's t -test, * P <0.05). ( d , e ) Representative western blot images ( d ) and quantification ( e ) of BNDF expression in brain homogenates from pathologically confirmed AD patients and age-matched HE (mean±s.e.m., n= 12, Student's t -test, * P <0.05). ( f , g ) BDNF levels in serum ( f ) and brain homogenates ( g ) of P301L tau transgenic pR5 mice and their wild-type (Wt) littermate controls at different ages (mean±s.e.m., n= 6 per age and per type, two-way analysis of variance (ANOVA), Tukey's test, ** P <0.01). ( h , i ) Representative western blot images ( h ) and quantification ( i ) of mouse BDNF in brain homogenates of 4-month-old mice (mean±s.e.m., n= 6 per age and per type, Student's t- test, * P <0.05). 4mo P301L denotes 4-month old P301L transgenic mouse; 4mo Wt denotes 4-month-old Wt littermates. ( j , k ) Representative western blot images ( j ) and quantification ( k ) of mouse BDNF in brain homogenates of 12-month-old mice (mean±s.e.m., n= 6 per age and per type, Student's t- test, * P <0.05, ** P <0.01).

Article Snippet: The homogenates were centrifuged at 10 000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa).

Techniques: Derivative Assay, Transgenic Assay, Comparison, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Translational Psychiatry

Article Title: Brain-derived neurotrophic factor protects against tau-related neurodegeneration of Alzheimer's disease

doi: 10.1038/tp.2016.186

Figure Lengend Snippet: Wide and stable expression of human brain-derived neurotrophic factor (BDNF) after intraventricular injection of adeno-associated virus carrying BDNF (AAV-BDNF). ( a ) Schematics of the construction and characterization of AAV-green fluorescent protein (AAV-GFP; left panel) and AAV-BDNF (right panel) vectors. ( b , c ) In order to exclude endogenous mouse BDNF, we applied a specific anti-human BDNF antibody to detect the expression of human BDNF within P301L mouse brain. Human BDNF expression is detectable in multiple brain regions ( b ), and GFP or human BDNF was selectively expressed in neurons but not in microglia (CD45-positive) or astrocytes (glial fibrillary acidic protein (GFAP)-positive) ( c ). Scale bar=80μm ( b ) or 40μm ( c ). ( d ) Comparison of human BDNF levels in brain homogenates of P301L mice treated with AAV-BDNF or AAV-GFP at 3 weeks (3w), 6 weeks (6w), 9 weeks (9w) and 9 months (9 m) after AAV injection initiated at 3 months of age ( n= 4 per group for 3w, 6w and 9w; n= 6 for 9 m group; mean±s.e.m.; two-way analysis of variance (ANOVA), Tukey's test, ** P <0.01). The concentrations of human BDNF were determined using specific human BDNF enzyme-linked immunosorbent assay (ELISA) kits. ( e ) Western analysis of total BDNF expression (including endogenous mouse BDNF and exogenous human BDNF) in brain homogenates ( n= 5 per group; mean±s.e.m., one-way ANOVA, Tukey's test, ** P <0.05; n.s. denotes no significant difference). 12mo P301L (AAV-BDNF) and 12mo P301L (AAV-GFP) denote 12-month-old P301L mice treated with AAV-BDNF and AAV-GFP, respectively; 12mo P301L (untreated) denotes untreated 12-month-old P301L mice; 12mo Wt denotes age-matched wild-type littermates.

Article Snippet: The homogenates were centrifuged at 10 000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa).

Techniques: Expressing, Derivative Assay, Injection, Virus, Comparison, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Translational Psychiatry

Article Title: Brain-derived neurotrophic factor protects against tau-related neurodegeneration of Alzheimer's disease

doi: 10.1038/tp.2016.186

Figure Lengend Snippet: Delivery of adeno-associated virus carrying brain-derived neurotrophic factor (AAV-BDNF) rescues the behavioral impairment of P301L mice. ( a ) Time schedule of behavioral tests. ( b , c ) The alternation percentage ( b ) and total number of entries ( c ) in the spontaneous alternation test (mean±s.e.m.; n= 8 per group, one-way analysis of variance (ANOVA), Tukey's test, * P <0.05). ( d ) Comparison of the percentages of novel arm entries among different groups (mean±s.e.m.; n= 8 per group, one-way ANOVA, Tukey's test, * P <0.05, ** P <0.01). ( e ) Comparison of the preference index in the novel object recognition test among different groups (mean±s.e.m.; n= 8 per group, one-way ANOVA, Tukey's test, * P <0.05). ( f ) Escape latency (seconds) during platform trials in Morris water maze (mean±s.e.m., n= 8 per group, two-way ANOVA, Tukey's test, * P <0.05). GFP, green fluorescent protein; Wt, wild-type.

Article Snippet: The homogenates were centrifuged at 10 000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa).

Techniques: Virus, Derivative Assay, Comparison

Journal: Translational Psychiatry

Article Title: Brain-derived neurotrophic factor protects against tau-related neurodegeneration of Alzheimer's disease

doi: 10.1038/tp.2016.186

Figure Lengend Snippet: Neuroprotective effects of adeno-associated virus carrying brain-derived neurotrophic factor (AAV-BDNF) on neurons. ( a , b ) Representative images ( a ) and quantitative statistics ( b ) of neurons in the CA1 region of the hippocampus of P301L mice (untreated, AAV-BDNF treated and AAV-green fluorescent protein (AAV-GFP) treated) and wild-type (Wt) littermates, using NeuN immunofluorescence (IF) assays (six mice per group; mean±s.e.m., one-way analysis of variance (ANOVA), Tukey's test, * P <0.05, ** P <0.01). Scale bar=25μm. ( c ) Schematic of basal forebrain in the coronal plane (upper-left panel) and representative images of choline acetyltransferase (ChAT)-positive cholinergic neurons in the medial septum (MS), intermediate part of the lateral septum (LSI) and vertical limb of the diagonal band (VDB) in AAV-BDNF and AAV-GFP groups. The inset shows the representative morphology of cholinergic neurons in MS at a higher magnification. V, ventral; D, dorsal; L, left; R, right. Scale bar=25μm. ( d ) Comparison of numbers of cholinergic neurons in MS, LSI and VDB between the two groups (six mice per group; mean±s.e.m., Student's t -test, * P <0.05, ** P <0.01). ( e ) Representative electron micrographs of pyramidal neurons from the CA1 region of the hippocampus in the four groups. It is worth noticing that in 12mo P301L (AAV-GFP) group and untreated P301L group, a number of pyramidal cells shows structural changes typical of apoptosis, including an enlarged perinuclear space, chromatic agglutination and karyopyknosis, whereas apoptosis-like cells were rarely found in 12mo P301L (AAV-BDNF) group, and pyramidal cells in this group usually displayed with uniform chromatin density and a large nucleus, which was similar to 12mo Wt group. Scale bar=2μm.

Article Snippet: The homogenates were centrifuged at 10 000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa).

Techniques: Virus, Derivative Assay, Immunofluorescence, Comparison, Agglutination

Journal: Translational Psychiatry

Article Title: Brain-derived neurotrophic factor protects against tau-related neurodegeneration of Alzheimer's disease

doi: 10.1038/tp.2016.186

Figure Lengend Snippet: Supportive effects of adeno-associated virus carrying brain-derived neurotrophic factor (AAV-BDNF) on neurogenesis and synapse. ( a , b ) Representative western blot images and quantitative analyses of the expressions of nestin, neuron-specific enolase (NSE) and doublecortin (DCX) in olfactory bulb ( a ) and dentate gyrus ( b ) of the four groups ( n= 6 per group, mean±s.e.m., one-way analysis of variance (ANOVA), Tukey's test, * P <0.05, ** P <0.01). ( c ) Representative western blot images and quantitative analyses for PSD-95, PSD-93, Syn-1, SNAP-25 and VAMP-1 expression in brain homogenates of the four groups ( n= 6 per group, mean±s.e.m., one-way ANOVA, Tukey's test, * P <0.05, ** P <0.01).

Article Snippet: The homogenates were centrifuged at 10 000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa).

Techniques: Virus, Derivative Assay, Western Blot, Expressing

Journal: Translational Psychiatry

Article Title: Brain-derived neurotrophic factor protects against tau-related neurodegeneration of Alzheimer's disease

doi: 10.1038/tp.2016.186

Figure Lengend Snippet: Genetic delivery of human brain-derived neurotrophic factor (BDNF) does not affect tau hyperphosphorylation. ( a ) Representative images of human pS396-tau immunostaining in the hippocampus and thalamus of different groups. Scale bar=100 μm. Insets show the representative morphology of pS396-tau-positive cells at higher magnification. ( b ) Comparison of the area fraction of human pS396-tau-positive immunostaining in the hippocampus (right panel) and thalamus (left panel) among the four groups ( n= 6 per group; mean±s.e.m., one-way analysis of variance (ANOVA), Tukey's test, ** P <0.01). No differences were found among 12mo P301L (untreated), 12mo P301L (AAV-green fluorescent protein, AAV-GFP) and 12mo P301L (AAV-BDNF, adeno-associated virus carrying BDNF). ( c ) Representative western blot images and quantitative analyses of tau hyperphosphorylation levels for multiple sites in 12mo P301L (AAV-GFP) and 12mo P301L (AAV-BDNF) groups, including pS396, pS202/T025 and pS404 ( n= 6 per group, mean±s.e.m., Student's t -test, no difference was found between the two groups). ( d ) Quantification of human pT181 (right panel) and human pT231 (left panel) levels in Tris buffer solution (TBS) and sodium dodecyl sulfonate (SDS) extracts of P301L brain tissue homogenates from AAV-GFP and AAV-BDNF groups ( n= 6 per group, mean±s.e.m., Student's t -test, no difference was found between two groups). ( e ) Representative western blot images and quantitative analyses of the Tau phosphorylation kinase GSK3beta and the Tau phosphatase PP2A in brain tissue homogenates of P301L mice from two groups ( n= 6 per group, mean±s.e.m., Student's t -test, no difference was found between two groups). AAV, adeno-associated virus.

Article Snippet: The homogenates were centrifuged at 10 000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa).

Techniques: Derivative Assay, Immunostaining, Comparison, Virus, Western Blot, Phospho-proteomics

Journal: Frontiers in Pharmacology

Article Title: Tibetan mineral-herbal medicine Zuotai alleviates the depressive-like behaviors in chronic restraint-stressed mice while regulating stress hormone, inflammation and monoamine

doi: 10.3389/fphar.2023.1098378

Figure Lengend Snippet:

Article Snippet: Soluble starch (AR, Sinopharm Chemical, Shanghai, China), imipramine hydrochloride (113-52-0, 98%; 3B Scientific, Hamburg, Germany), sucrose (AR, Tianjin Beichen Fangzheng Reagent Factory, Tianjian, China), paraformaldehyde (AR, Tianjin Guangfu Institute of Fine Chemical Industry, Tianjin, China), mouse IL-6 ELISA kits (85-BMS603/2; eBioscience, CA, USA), mouse IL-6 (ELM-IL6-1, Raybiotech, GA, USA), mouse IL-1β ELISA kits (85-BMS6002; eBioscience, CA, USA), mouse IL-1β (ELM-IL1b-1, Raybiotech, GA, USA), mouse TNF-α ELISA kits (85-BMS607/3, eBioscience, CA, USA), and mouse TNF-α (ELM-TNFa-1, Raybiotech, GA, USA), norepinephrine enzyme ELISA kits (CEA907Ge; Cloud-Clone, Wuhan, China), serotonin ELISA kits (CEA808Ge; Cloud-Clone, Wuhan, China), brain-derived neurotrophic factor (BDNF) ELISA kits (SEA011Mu; Cloud-Clone, Wuhan, China), Corticosterone (Cort) ELISA kits (CSB-E07969m, CUSABIO, Wuhan China), total mercury standard solution (1 mg/mL, GSB04-1729-2004; National Nonferrous Metals and Electronic Materials Analysis and Testing Center, China), nitric acid (GR, Baiyin Liangyou Chemical Reagent Company, Gansu, China), high purity oxygen (purity ≥99.99%, Xining Laoqian Gas Trading Company, China).

Techniques: Suspension, Solubility, X-ray Diffraction, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Standard Deviation

Journal: NPJ Vaccines

Article Title: A multi-targeting immunotherapy ameliorates multiple facets of Alzheimer’s disease in 3xTg mice

doi: 10.1038/s41541-024-00942-9

Figure Lengend Snippet: a Methods for pathological evaluation: Outline of methods for evaluating mouse brain pathology after vaccine immunization. b Aβ Immunostaining: 6E10 immunostaining for Aβ burden in 3xTg mice brains. (Black scale bar: 100 μm). c Quantification of Aβ immunostaining: Quantification of 6E10 immunostaining in the hippocampal CA1 and cortex regions. d Aβ42 analysis: Aβ42 levels in soluble and insoluble brain homogenates by flow cytometry (The experimental samples are mixed samples ( n = 9–10) and repeated three times). e pTau Immunostaining: AT8 immunostaining for phosphorylated-tau in 3xTg mice brains. (Red scale bar: 50 μm). f Quantification of pTau immunostaining: Quantification of AT8 immunostaining in the hippocampal CA1 and cortex regions ( n = 9–10). g pTau202/205 analysis: pTau202/205 levels in soluble and insoluble brain homogenates by western blot (Mixed samples, n = 9–10). GAPDH served as the internal control. The relative content of each sample is marked with a red number. h PHF-13 Immunostaining: PHF-13 immunostaining for phosphorylated-tau in 3xTg mice brains. (Red scale bar: 50 μm). i Quantification of PHF-13 immunostaining: Quantification of PHF-13 immunostaining in the hippocampal DG and cortex regions ( n = 9–10). j pTau396 analysis: pTau396 levels in soluble and insoluble brain homogenates by western blot (Mixed samples, n = 9–10). GAPDH served as the internal control. The relative content of each sample is marked with a red number. k Phospho-tau (Ser404) immunostaining: Immunostaining for phosphorylated-tau (Ser404) in 3xTg mice brains. (Red scale bar: 50 μm). l Quantification of phospho-tau (Ser404) immunostaining: Quantification of phospho-tau (Ser404) immunostaining in hippocampal CA1 and cortex regions ( n = 9–10). m pTau404 analysis: pTau404 levels in soluble and insoluble brain homogenates by western blot (Mixed samples, n = 9–10). GAPDH served as the internal control. The relative content of each sample is marked with a red number. *Data presented as mean ± SEM. Multivariate ANOVA was employed when p ≤ 0.05. Bonferroni-corrected t -test was otherwise applied. Statistical significance denoted as * p < 0.05, ** p < 0.01, *** p < 0.001; ns not significant. (Fig. 2a was created with BioRender.com).

Article Snippet: The levels of Aβ42 in serum and brain homogenates (both soluble and insoluble fractions) of 3xTg mice were determined using Human Aβ42 ELISA kits (Uscn Life Science Inc., China), following the manufacturer’s protocol.”

Techniques: Immunostaining, Flow Cytometry, Western Blot, Control

Journal: NPJ Vaccines

Article Title: A multi-targeting immunotherapy ameliorates multiple facets of Alzheimer’s disease in 3xTg mice

doi: 10.1038/s41541-024-00942-9

Figure Lengend Snippet: a 6E10 immunostaining for Aβ burden: Immunostaining for Aβ burden in the brains of 3xTg mice treated with vaccines or vehicle. (White scale bar: 250 μm). b Quantification of Aβ immunostaining: quantification of 6E10 immunostaining in the hippocampus and cortex regions ( n = 13–15). c , d Aβ42 analysis: Aβ42 levels in soluble and insoluble brain homogenates in the hippocampus. ( c ) and cortex regions ( d ) by flow cytometry (The experimental samples are mixed samples ( n = 13–15) and repeated three times). e PHF-13 Immunostaining for pTau: Immunostaining for phosphorylated-tau (PHF-13) in the brains of 3xTg mice treated with vaccines or vehicle (Red scale bar: 50 μm). f Quantification of PHF-13 immunostaining: Quantification of PHF-13 immunostaining in the hippocampal DG and Cortex regions ( n = 13–15). g , h pTau396 analysis: pTau396 levels in soluble and insoluble brain homogenates in the hippocampal DG ( g ) and Cortex regions ( h ) by western blot ( n = 13–15). GAPDH served as the internal control. The relative content of each sample is marked with a red number. i Hindlimb clamping (Clasp Score): Clasp scores in hindlimb clamping ( n = 13–15 per group, one-way ANOVA). j Novel object recognition (NOR): Cognitive functions assessed in novel object recognition ( n = 13–15 per group, one-way ANOVA). k Morris water maze—escape latency: Escape latency to the platform during the training trials in the Morris water maze ( n = 13–15 per group, two-way ANOVA). l Morris water maze—platform crossings: Numbers of platform location crossings in the probe trial of Morris water maze ( n = 13–15 per group, one-way ANOVA). *Data were mean ± SEM. Multivariate ANOVA was employed when p ≤ 0.05. Bonferroni-corrected t -test was otherwise applied. * p < 0.05, ** p < 0.01, *** p < 0.001; ns not significant.

Article Snippet: The levels of Aβ42 in serum and brain homogenates (both soluble and insoluble fractions) of 3xTg mice were determined using Human Aβ42 ELISA kits (Uscn Life Science Inc., China), following the manufacturer’s protocol.”

Techniques: Immunostaining, Vaccines, Flow Cytometry, Western Blot, Control

Journal: NPJ Vaccines

Article Title: A multi-targeting immunotherapy ameliorates multiple facets of Alzheimer’s disease in 3xTg mice

doi: 10.1038/s41541-024-00942-9

Figure Lengend Snippet: a GFAP immunostaining in the hippocampus of premorbid 3xTg mice treated with the dual vaccine or PP vehicle (Vector). (White scale bar: 250 μm, Red scale bar: 50 μm). b Quantification of GFAP immunostaining in the hippocampus. c Representative Iba-1 images and microglia quantification: Representative Iba-1 images and quantification of branching and endpoint of microglia in the hippocampus. n = 7 mice per genotype with 6–8 microglia quantified per mouse. (Black scale bar: 100 μm, Red scale bar: 50 μm) d , e Microglial morphology analysis: Quantitative assessment of branching ( d ) and endpoint ( e ) of microglia. f Cytokine levels in brain lysates: Levels of TNF-α (left) and IL-1β (right) in the brain lysates of 3xTg mice determined using corresponding ELISA kits. ( n = 6–7). The experimental samples were mixed ( n = 6–7) and repeated three times. g , h Western blot analysis: Western blot analysis of GFAP, Iba-1, and GAPDH in the brain lysates of 3xTg mice treated with the dual vaccine or vehicle. GAPDH served as the internal control. The relative content of each sample is marked with a red number. *Data presented as mean ± SEM. Multivariate ANOVA was employed when p ≤ 0.05. Bonferroni-corrected t -test was otherwise applied. Statistical significance denoted as * p < 0.05, ** p < 0.01, *** p < 0.001; ns not significant.

Article Snippet: The levels of Aβ42 in serum and brain homogenates (both soluble and insoluble fractions) of 3xTg mice were determined using Human Aβ42 ELISA kits (Uscn Life Science Inc., China), following the manufacturer’s protocol.”

Techniques: Immunostaining, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: NPJ Vaccines

Article Title: A multi-targeting immunotherapy ameliorates multiple facets of Alzheimer’s disease in 3xTg mice

doi: 10.1038/s41541-024-00942-9

Figure Lengend Snippet: a , b Antibody concentrations in the brain: Concentrations of antibodies against Aβ and individual phosphorylated Tau sites in the brain of 3xTg mice from the premorbid group ( a ) and onset group ( b ). c , d Vaccine-generated antibodies impact on Aβ42: Vaccine-generated antibodies entering the brain and altering the Aβ42 content in the brain and peripheral blood of mice after immunization in premorbid group ( c ) and onset group ( d ). *Data presented as mean ± SEM. Multivariate ANOVA was employed when p ≤ 0.05. Bonferroni-corrected t -test was otherwise applied. Statistical significance denoted as * p < 0.05, ** p < 0.01, *** p < 0.001; ns not significant.

Article Snippet: The levels of Aβ42 in serum and brain homogenates (both soluble and insoluble fractions) of 3xTg mice were determined using Human Aβ42 ELISA kits (Uscn Life Science Inc., China), following the manufacturer’s protocol.”

Techniques: Generated