RhoA/Rock2/Limk1/cofilin1 pathway is involved in attenuation of neuronal dendritic spine loss by paeonol in the frontal cortex of D-galactose and aluminum­induced Alzheimer's disease­like rat model.

Alzheimer's disease (AD) has become the most prevalent neurodegenerative disorder. Given the pathogenesis of AD is unclear, there is currently no drug approved to halt or delay the progression of AD. Therefore, it is pressing to explore new targets and drugs for AD. In China, polyphenolic Chinese herbal medicine has been used for thousands of years in clinical application, and no toxic effects have been reported. In the present study, using D­galactose and aluminum­induced rat model, the effects of paeonol on AD were validated via the Morris water maze test, open field test, and elevated plus maze test. Neuronal morphology in frontal cortex was assessed using ImageJ's Sholl plugin and RESCONSTRUCT software. RhoA/Rock2/Limk1/cofilin1 signaling pathway­related molecules were determined by Western blotting. Cofilin1 and p­cofilin1 were analyzed by immunofluorescence. Results showed that pre­treatment with paeonol attenuated D­galactose and aluminum­induced behavioral dysfunction and AD­like pathological alterations in the frontal cortex. Accompanied by these changes were the alterations in the dendrite and dendritic spine densities, especially the mushroom­type and filopodia­type spines in the apical dendrites, as well as actin filaments. In addition, the activity and intracellular distribution of cofilin1 and the molecules RhoA/Rock2/Limk1 that regulate the signaling pathway for cofilin1 phosphorylation have also changed. Our data suggests that paeonol may be through reducing Aß levels to alleviate the loss of fibrillar actin and dendrites and dendritic spines via the Rho/Rock2/Limk1/cofilin1 signaling pathway in the frontal cortex, and ultimately improving AD­like behavior.

Novel antidepressant effects of Paeonol alleviate neuronal injury with concomitant alterations in BDNF, Rac1 and RhoA levels in chronic unpredictable mild stress rats.

RATIONALE: Increasing evidence has suggested that major depressive disorder (MDD) is highly associated with brain-derived neurotrophic factor (BDNF) levels, dendrites atrophy, and loss of dendritic spines, especially in emotion-associated brain regions including the hippocampus. Paeonol is a kind of polyphenols natural product with a variety of therapeutic effects. Recent studies have reported its antidepressant effects. However, it is unclear what signaling pathways contribute to improve MDD. OBJECTIVE: The present study investigated the effect of Paeonol on hippocampal neuronal morphology and its possible signaling pathways in chronic unpredictable mild stress (CUMS) rat model. METHODS: Using CUMS rat model, the antidepressant-like effect of Paeonol was validated via depression-related behavioral tests. Neuronal morphology in hippocampal CA1 and DG was assessed using ImageJ's Sholl plugin and RESCONSTRUCT software. BDNF signaling pathway-related molecules was determined by Western blotting. RESULTS: Paeonol attenuated CUMS-induced depression-like behaviors, which were accompanied by hippocampal neuronal morphological alterations. After Paeonol treatment for 4 weeks, the dendritic length and complexity and the density of dendritic spines markedly increased in the hippocampal CA1 and the dentate gyrus (DG). However, CUMS or Paeonol treatment does not selectively affect dendritic spine types. Simultaneously, administration of Paeonol deterred CUMS-induced cofilin1 activation that is essential for remolding of dendritic spines. The induction of CUMS downregulated BDNF levels and upregulated Rac1/RhoA levels; however, the tendency of these was inhibited by treatment with Paeonol. CONCLUSION: Our data suggest that BDNF-Rac1/RhoA pathway may be involved in attenuation of CUMS-induced behavioral and neuronal damage by Paeonol that may represent a novel therapeutic agent for depression.

Novel derivative of Paeonol, Paeononlsilatie sodium, alleviates behavioral damage and hippocampal dendritic injury in Alzheimer's disease concurrent with cofilin1/phosphorylated-cofilin1 and RAC1/CDC42 alterations in rats.

Alzheimer's disease (AD) is a typical hippocampal amnesia and the most common senile dementia. Many studies suggest that cognitive impairments are more closely correlated with synaptic loss than the burden of amyloid deposits in AD progression. To date, there is no effective treatment for this disease. Paeonol has been widely employed in traditional Chinese medicine. This compound improves learning behavior in an animal model; however, the mechanism remains unclear. In this study, Paeononlsilatie sodium (Pa), a derivative of Paeonol, attenuated D-galactose (D-gal) and AlCl3-induced behavioral damages in rats based on evaluations of the open field test (OFT), elevated plus maze test (EPMT), and Morris water maze test (MWMT). Pa increased the dendritic complexity and the density of dendritic spines. Correlation analysis indicated that morphological changes in neuronal dendrites are closely correlated with behavioral changes. Pa treatment reduced the production of Aß, affected the phosphorylation and redistribution of cofilin1 and inhibited rod-like formation in hippocampal neurons. The induction of D-gal and AlCl3 promoted the expression of RAC1/CDC42 expression; however, the tendency of gene expression was inhibited by pretreatment with Pa. Taken together, our results suggest that Pa may represent a novel therapeutic agent for the improvement of cognitive and emotional behaviors and dendritic morphology in an AD animal model.

Expression and functional regulation of stemness gene Lgr5 in esophageal squamous cell carcinoma.

Cancer stem cells (CSCs) are defined as a rare subpopulation of undifferentiated cells with biological characteristics that include the capacity for self-renewal, differentiation into various lineages, and tumor initiation. To explore the mechanism of CSCs in esophageal squamous cell carcinoma (ESCC), we focused on Leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5), a target gene of the Wnt signaling pathway, which has been identified as a marker of intestinal stem cells and shown to be overexpressed in several human malignancies. Lgr5 expression was significantly correlated with lymph node metastasis, increased depth of invasion, increased tumor size, advanced differentiation, higher AJCC stage and poorer survival. Silencing of Lgr5 expression in the ESCC cell line KYSE450 by small interfering RNA (siRNA) strongly inhibited cell proliferation, migration and invasion ability, the expression of CSCs-related genes and Wnt/ß-catenin signaling. In addition, Lgr5 was highly expressed in ESCC spheroid body cells, which were identified by high expression of CSCs-related genes, and high tumorigenicity in vivo. Taken together, these results demonstrate that Lgr5 activation of Wnt/ß-catenin signaling is a potential mechanism to promote the progression of ESCC and ESCC stem cell renewal, and Lgr5 may be used as a molecular target for the development of treatments for ESCC.

[Comparison of four diagnostic criteria for idiopathic inflammatory myopathy].

OBJECTIVE: To compare the clinical values of four different criteria for diagnosing idiopathic inflammatory myopathy (IIM). METHODS: The four different criteria published in 1975, 1995, 1997 and 2004 were applied to 94 IIM patients and 98 patients with other myopathies in West China Hospital. RESULTS: The sensitivity of the four criteria for diagnosing IIM was 56. 4% (1975), 87. 2% (1995), 61.7 % (1997), and 52. 1% (2004) respectively. The specificity of the four criteria for diagnosing IIM was 78. 6% (1975), 20. 4% (1995), 78. 6% (1997), and 90. 8% (2004) respectively. The Youden index was 35.0% (1975), 7.6% (1995), 40. 3% (1997), and 42.9% (2004), with an odd product of 4. 74 (1975), 1. 75 (1995), 5. 91 (1997) and 10. 77 (2004) respectively. The Kappa value of the four criteria was 0. 351 (1975, P<0.05), 0. 075 (1995, P>0. 05), 0. 404 (1997, P<0.05 ) and 0. 433 (2004, P<0.05), and their area under the ROC curve was 0. 675 (1975, P=0. 00), 0. 538 (1995, P=0.36),0. 701 (1997, P=0.00) and 0. 715 (2004, P=0.00) respectively. CONCLUSION: The 2004 criteria have a better value in diagnosing IIM.

[Levels of acute-phase-reactants in patient with dermatomysitis and its correlations with IL-6 and dermatomyositis disease activity].

OBJECTIVE: To determine the association between acute-phase-reactants (APR) and interleukin-6 (IL-6) in patient with dermatomysitis (DM). METHODS: The levels of C-reactive protein (CRP), serum amyloid A protein(SAA) and serum ferritin (SF) in peripheral blood of 31 adult DM patients were determined by chemiluminescence immunoassay, and compared with those of 23 patients with systemic lupus erythematosus (SLE), 22 patients with rheumatoid arthritis (RA), and 18 patients with Sjagren syndrome (SS). The correlations between the levels of those APR and IL-6 were examined. We also measured dermatomyositis disease activity using myositis disease activity assessment tool (MDAAT), and examined its association with APR levels. RESULTS: DM patients had significantly lower level of CRP [(17. 08 +/- 17. 18) mg/L] than those patients with RA [(85. 95 +/-60.62) mg/L, P<0. 000 1], SLE [(51. 34+/-52. 98) mg/L, P=0. 006] and SS [(47. 00+/-47. 24) mg/L, P= 0.018]. DM patients had significantly lower level of SAA [(92. 04 +/- 98. 93) mg/L] than those patients with RA [(311.30 +/- 292. 45) mg/L, P= 0. 002] and SS [(284. 31 +/- 325. 30) mg/L, P= 0. 025]. DM patients had significantly higher level of SF [(510. 10 +/- 610. 73) ng/mL] than those patients with SS [(220. 33 +/- 164. 07) ng/ mL, P=0. 02], as well as those with RA and SLE albeit without statistical significance. All of the three APRs were positive correlated with IL-6 level. No significant associations between APR and systemic or global disease activities were found, although CRP was associated with constitutional disease activity and SF was associated with pulmonary disease activity. CONCLUSION: DM patients have lower levels of elevated APR than the other three common connective tissue diseases, which is associated with IL-6 but not with global disease activity.

[The effect of genetic polymorphism of the exon 2 of the beta-lactoglobulin gene on the milk composition in Chinese Holstein].

OBJECTIVE: In order to study the effect of the polymorphism at the exon2 region of the (3-LG allele gene on milk composition and yield. METHODS: The single-strand conformation polymorphism method (PCR-SSCP) was used to analyze for polymorphism the exon2 region of the 3-LG gene (NCBI accession number: DQ489319) in Chinese Holstein. RESULTS: Eight SSCP patterns were detected in the fragments: ab, abc, abd, abe, abcd, abce, abde and abcde, and the patterns frequencies as follows: 0.14, 0.10, 0.27, 0.23, 0.05, 0.04, 0.11 and 0.06 (P < 0.05); Six single nucleotide polymorphism (SNP) were detected in this study: sitel C>T, site2 T>C, site3 C>T, site4 C>C, site5 C> A, site6 A>T or C, and the polymorphism infonnation content (PIC) of these SNPs were in median or high polymorphism (PIC > 0.25). CONCLUSION: These SNPs at the exon2 region of the beta-LG gene were remarkably and affected milk performance traits (milk yield, protein and fat contents) in Chinese Holstein.