Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Effect of Sirtuin-1 on Synaptic Plasticity in Nucleus Accumbens in a Rat Model of Heroin Addiction

doi: 10.12659/MSM.910550

Figure Lengend Snippet: Cells positive for SIRT1, Cdk5, NF-κB, FOXO1, Cbp, and Syn in the NAc region (×400) and their optical density values ( A ) Saline control (SC) group; ( B ) Heroin-addicted (HA) group; ( C ) AAV-rSirt1 group; ( D ) AAV-rSirt1 shRNA group. # P <0.05 compared with the SC group; * P <0.05 compared with the HA group.

Article Snippet: The reagents and analysis systems used were as follows: heroin (purity 92.09%, provided by the Guizhou Public Security Bureau), pentobarbital sodium (Sigma, USA), naloxone hydrochloride (Chongqing YaoPharma, China), mouse monoclonal anti-SIRT1 (Abcam, USA), rabbit polyclonal anti-Cdk5, rabbit polyclonal anti-NF-κB (Bioss, China), rabbit monoclonal anti-FOXO1 (Abcam, USA), rabbit polyclonal anti-Cbp (Bioss, China), rabbit monoclonal anti-PSD95 (Abcam, USA), rabbit monoclonal anti-Syn (Abcam, USA), mouse monoclonal anti-beta-actin (Abcam, USA), SuperScript III RT reverse transcription kit (Invitrogen, USA) Sybr qPCR mix (Invitrogen, USA), Plasmid Mini kit (TransGen Biotech, Beijing, China), gel extraction kit (Omega), Trans2K Plus II DNA Marker (TransGen, Beijing, China), DNA primers (Invitrogen, USA), DNA sequencing (Invitrogen, USA), restriction endonuclease EcoRI (NEB, USA), T4 DNA Ligase (NEB, USA), Plasmid Maxi Kit (Qiagen, Germany), CPT High-efficiency Transfection Kit (Virotherapy Technologies, Wuhan, China).

Techniques: shRNA

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Effect of Sirtuin-1 on Synaptic Plasticity in Nucleus Accumbens in a Rat Model of Heroin Addiction

doi: 10.12659/MSM.910550

Figure Lengend Snippet: Detection of SIRT1, Cdk5, FOXO1, NF-κB, Syn, and PSD95 protein levels in each group by immunoblotting. # P <0.05 compared with the saline control group; * P <0.05 compared with the HA group .

Article Snippet: The reagents and analysis systems used were as follows: heroin (purity 92.09%, provided by the Guizhou Public Security Bureau), pentobarbital sodium (Sigma, USA), naloxone hydrochloride (Chongqing YaoPharma, China), mouse monoclonal anti-SIRT1 (Abcam, USA), rabbit polyclonal anti-Cdk5, rabbit polyclonal anti-NF-κB (Bioss, China), rabbit monoclonal anti-FOXO1 (Abcam, USA), rabbit polyclonal anti-Cbp (Bioss, China), rabbit monoclonal anti-PSD95 (Abcam, USA), rabbit monoclonal anti-Syn (Abcam, USA), mouse monoclonal anti-beta-actin (Abcam, USA), SuperScript III RT reverse transcription kit (Invitrogen, USA) Sybr qPCR mix (Invitrogen, USA), Plasmid Mini kit (TransGen Biotech, Beijing, China), gel extraction kit (Omega), Trans2K Plus II DNA Marker (TransGen, Beijing, China), DNA primers (Invitrogen, USA), DNA sequencing (Invitrogen, USA), restriction endonuclease EcoRI (NEB, USA), T4 DNA Ligase (NEB, USA), Plasmid Maxi Kit (Qiagen, Germany), CPT High-efficiency Transfection Kit (Virotherapy Technologies, Wuhan, China).

Techniques: Western Blot

Journal: CNS Neuroscience & Therapeutics

Article Title: Surgical Stress Induces Brain‐Derived Neurotrophic Factor Reduction and Postoperative Cognitive Dysfunction Via Glucocorticoid Receptor Phosphorylation in Aged Mice

doi: 10.1111/cns.12368

Figure Lengend Snippet: Surgical stress elevated GR phosphorylation at ser220 by CDK5 and its regulator in prefrontal cortex of POCD mice. (A), GR phosphorylation was tested with Western blot. The phosphorylation at ser220 site was significantly elevated in POCD group (two‐tailed unpaired t‐test, P = 0.0136 for pGRs220; P = 0.2303 for pGRs212; P = 0.3860 for pGRs234), compared with control group. (B) The level of CDK5 and its regulators p35, p25 in prefrontal cortex was tested with Western blot in POCD and control mice. The level of CDK5 was upregulated in POCD group compared with control (two‐tailed unpaired t‐test, P = 0.0178). CDK5 regulators p35 (P = 0.0140), p25 (P = 0.0139) also elevated in prefrontal cortex compared with control. (C), nonparametric correlation analysis of GR phosphorylation and behavioral index in 20‐month groups (CDK5 with MWM, r2 = 0.75, P < 0.001; CDK5 with EPM, r2 = 0.78, P < 0.001; pGRs220 with MWM, r2 = 0.63, P < 0.001; pGRs220 with EPM, r2 = 0.73, P < 0.001) (C). (*P < 0.05, POCD compared with control) (n = 8 mice/group). Error bar represents SEM. (D), no significant changes on pGRs220 (P = 0.7576), CDK5 (P = 0.8640) and its regulators (P = 0.8182 for p35 and P = 0.8844 for p25) in 6‐month groups. (*P < 0.05, POCD compared with control) (n = 8 mice/group). Data are Mean ± SEM (Error bars).

Article Snippet: After transferred onto PVDF membrane, the following primary antibodies were used: sheep anti‐BDNF antibody (1:5000, millipore, Billerica, MA, USA); rabbit anti‐GR and anti‐p‐GRs220 antibody (1:12000, CST, Boston, MA, USA); rabbit anti‐CDK5 antibody (1:5000, BD, NJ, USA); rabbit anti‐p35 antibody (1:5000, BD); rabbit anti‐p‐GRs212 (1:2000, Anbo, San Francisco, CA, USA); and rabbit anti‐p‐GRs234 (1:12000, CST).

Techniques: Western Blot, Two Tailed Test

Journal: CNS Neuroscience & Therapeutics

Article Title: Surgical Stress Induces Brain‐Derived Neurotrophic Factor Reduction and Postoperative Cognitive Dysfunction Via Glucocorticoid Receptor Phosphorylation in Aged Mice

doi: 10.1111/cns.12368

Figure Lengend Snippet: Surgical stress‐induced GR phosphorylation, BDNF reduction and cognitive dysfunction are rescued by CDK5 inhibitor. (A) Roscovitine, a specific CDK5 inhibitor was treated after the surgery. The performance on Morris water maze and elevated plus maze was rescued with treatment of roscovitine. POCD+roscovitine group performed shorter latency to find the hidden platform (two‐way ANOVA, treatment condition × day, F(1,98) = 11.21, P < 0.001; POCD+roscovitine compared with POCD group followed by Tukey's post hoc test, P < 0.001; one‐way ANOVA following with Tukey's post hoc test was used to compare latency on each day, P < 0.05 on day 6 and 7, POCD+roscovitine compared with POCD) and spent longer time in target area with a removed platform (one‐way ANOVA following with Tukey's post hoc test, P = 0.0446), compared with POCD group. No significant difference on swimming speed among groups (P = 0.8075, POCD+roscovitine compared with POCD) and swimming paths were also presented. In elevated plus maze test, time spent in open arm was also rescued with roscovitine treatment (one‐way ANOVA following with Tukey's post hoc test, P = 0.0199, POCD+roscovitine compared with POCD). (B), the level of GR phosphorylation at ser220 site in nucleus was rescued in POCD+roscovitine group compared with POCD (P = 0.0277). The expression of BDNF level in prefrontal cortex was also rescued in POCD+roscovitine group compared with POCD (P = 0.0162). (*P < 0.05, POCD compared with control; # P < 0.05, POCD+roscovitine compared with POCD) (n = 8 mice/group). Data are Mean ± SEM (Error bars).

Article Snippet: After transferred onto PVDF membrane, the following primary antibodies were used: sheep anti‐BDNF antibody (1:5000, millipore, Billerica, MA, USA); rabbit anti‐GR and anti‐p‐GRs220 antibody (1:12000, CST, Boston, MA, USA); rabbit anti‐CDK5 antibody (1:5000, BD, NJ, USA); rabbit anti‐p35 antibody (1:5000, BD); rabbit anti‐p‐GRs212 (1:2000, Anbo, San Francisco, CA, USA); and rabbit anti‐p‐GRs234 (1:12000, CST).

Techniques: Expressing