IGFBP7 promotes gastric cancer by facilitating epithelial-mesenchymal transition of gastric cells.

Gastric cancer (GC) with high morbidity and mortality is one major cause of tumor-related death. Mechanisms underlying GC invasion and metastasis remain unclear. IGFBP7 exerted variable effects in different cancers and its role in GC is controversial. Here, IGFBP7 was found to be upregulated and elevated IGFBP7 expression represented a poorer overall survival in GC using bioinformatics analysis. Moreover, IGFBP7 was up-regulated in human GC specimens and promoted tumor growth in xenograft tumor animals. For GC cell lines, we found that IGFBP7 was also upregulated and facilitated the cell malignant behavior and EMT of GC cells, which may involve NF-κB and ERK signaling pathways. This research may provide new avenues for GC therapy.

Integrated proteomics and metabolomics analysis of D-pinitol function during hippocampal damage in streptozocin-induced aging-accelerated mice.

Purpose: Diabetes can cause hippocampal damage and lead to cognitive impairment. Diabetic cognitive impairment (DCI) is a chronic complication of diabetes associated with a high disability rate; however, its pathogenesis and therapeutic targets are unclear. We aimed to explore the mechanism of hippocampal damage during diabetes and evaluate the potential role of D-pinitol (DP) in protecting hippocampal tissue and improving cognitive dysfunction. Methods: DP (150 mg/kg/day) was administered intragastrically to streptozocin-induced aging-accelerated mice for 8 weeks. Hippocampal tissues were examined using tandem mass tag (TMT)-based proteomics and liquid chromatography-mass spectrometry (LC-MS)/MS-based non-targeted metabolomic analysis. Differentially expressed proteins (DEPs) and differentially regulated metabolites (DRMs) were screened for further analysis, and some DEPs were verified using western blotting. Results: Our results showed that 329 proteins had significantly altered hippocampal expression in untreated diabetic mice (DM), which was restored to normal after DP treatment in 72 cases. In total, 207 DRMs were identified in the DM group, and the expression of 32 DRMs was restored to normal post-DP treatment. These proteins and metabolites are involved in metabolic pathways (purine metabolism, arginine and proline metabolism, and histidine metabolism), actin cytoskeleton regulation, oxidative phosphorylation, and Rap1-mediated signaling. Conclusions: Our study may help to better understand the mechanism of diabetic hippocampal damage and cognitive impairment and suggest a potential therapeutic target.

Silencing of NOD2 protects against diabetic cardiomyopathy in a murine diabetes model.

The aim of the present study was to investigate the role of the nucleotide­binding oligomerization domain (NOD) 2 in high glucose (HG)­induced myocardial apoptosis and fibrosis in mice. Mouse models of diabetes were induced by streptozotocin (STZ). NOD2 expression was knocked down by injection of lentivirus­mediated short­hairpin RNA. Alternatively, small interfering RNA­NOD2 was transfected into cardiomyocytes and cardiac fibroblasts (CFs). A hemodynamic assay was used to assess the cardiac function in the mouse model. Hematoxylin and eosin, Masson and terminal deoxynucleotidyl transferase dUTP nick end labeling staining was performed to observe pathological changes and injury of myocardial tissue. The expression levels of NOD2, collagen I and III, and transforming growth factor­ß (TGF­ß) and apoptotic proteins were quantified by reverse transcription­quantitative polymerase chain reaction and western blotting. NOD2 silencing ameliorated diabetes­induced myocardial apoptosis and fibrosis in mice. NOD2, collagen I, collagen III, TGF­ß and pro­apoptotic proteins were upregulated in the diabetic cardiomyopathy (DCM) model group, but interference of NOD2 suppressed these alterations in protein expression levels. NOD2 is upregulated in HG­induced primary cardiomyocytes and CFs. Suppression of NOD2 attenuated HG­induced cardiomyocyte apoptosis and proliferation of CFs. Overall, NOD2 silencing alleviated myocardial apoptosis and fibrosis in diabetic mice. The results of the present study demonstrated an understanding of the role of NOD2 in diabetes­induced cardiomyopathy, which provides a novel target and therapies for the prevention and treatment of DCM.

FGF-16 protects against adverse cardiac remodeling in the infarct diabetic heart.

Till now, no functional studies for FGF-16 in diabetic heart have been reported. Therefore, this study aims to evaluate the potential function of FGF-16 in inhibiting adverse cardiac remodeling in post myocardial infarction (MI) of diabetic heart. We investigated the role of fibroblast growth factor-16 (FGF-16) in post-MI remodeling and role of cardio-protection in the diabetic infarct heart. Adult db/db diabetic mice were assigned to sham group, MI group and MI+FGF-16 group, respectively. MI group was induced by permanent coronary artery ligation, and the mice were subjected to 2D trans-thoracic echocardiography 2-4 weeks post-surgery. The results showed that the infiltration of monocytes, the associated pro-inflammatory cytokines were significantly increased, and the adverse cardiac remodeling and left ventricular dysfunction were observed in MI group. FGF-16 treatment protected against apoptosis, cardiac dysfunction and chamber dilatation post-MI, and decreased monocyte infiltration and cardiomyocyte hypertrophy/apoptosis. Meanwhile, the FGF-16 treatment also attenuated interstitial fibrosis and myocardial inflammation post-MI, increased M2 macrophage differentiation and associated anti-inflammatory cytokines, reduced adverse remodeling, and improved cardiac function. In conclusion, our results suggest that the heart appears to be a target of systemic and possibly locally generated FGF-16, which plays a therapeutic role in cardiac protection in the post-MI diabetic heart.

A Spontaneous Structural Transition of {U24 Pp12 } Clusters Triggered by Alkali Counterion Replacement in Dilute Solution.

A transition between two isomeric clusters involving the change of the main skeleton structure of a well-defined, rigid molecular cluster [(UO2 )24 (O2 )24 (P2 O7 )12 ]48- , {U24 Pp12 }, is achieved by simply introducing proper alkali cations into its dilute aqueous solution. While the unique structural transition can be triggered by introducing any of the Na+ /K+ /Rb+ /Cs+ alkali ions, the two isomers, Li/Na-{U24 Pp12 } and Na/K-{U24 Pp12 }, as typical macroions, can accurately choose among different alkali counter-cations based on their hydrated sizes, and the ion selectivity process clearly showed endothermic features. The preferred K+ and Rb+ ions have suitable sizes to be incorporated into the proper windows on {U24 Pp12 } nanocapsules, as supported by the transition points in both ITC studies and IR measurements.

Genetic Ablation of Type III Adenylyl Cyclase Exerts Region-Specific Effects on Cilia Architecture in the Mouse Nose.

We recently reported that olfactory sensory neurons in the dorsal zone of the mouse olfactory epithelium exhibit drastic location-dependent differences in cilia length. Furthermore, genetic ablation of type III adenylyl cyclase (ACIII), a key olfactory signaling protein and ubiquitous marker for primary cilia, disrupts the cilia length pattern and results in considerably shorter cilia, independent of odor-induced activity. Given the significant impact of ACIII on cilia length in the dorsal zone, we sought to further investigate the relationship between cilia length and ACIII level in various regions throughout the mouse olfactory epithelium. We employed whole-mount immunohistochemical staining to examine olfactory cilia morphology in phosphodiesterase (PDE) 1C-/-;PDE4A-/- (simplified as PDEs-/- hereafter) and ACIII-/- mice in which ACIII levels are reduced and ablated, respectively. As expected, PDEs-/- animals exhibit dramatically shorter cilia in the dorsal zone (i.e., where the cilia pattern is found), similar to our previous observation in ACIII-/- mice. Remarkably, in a region not included in our previous study, ACIII-/- animals (but not PDEs-/- mice) have dramatically elongated, comet-shaped cilia, as opposed to characteristic star-shaped olfactory cilia. Here, we reveal that genetic ablation of ACIII has drastic, location-dependent effects on cilia architecture in the mouse nose. These results add a new dimension to our current understanding of olfactory cilia structure and regional organization of the olfactory epithelium. Together, these findings have significant implications for both cilia and sensory biology.

An Olfactory Cilia Pattern in the Mammalian Nose Ensures High Sensitivity to Odors.

In many sensory organs, specialized receptors are strategically arranged to enhance detection sensitivity and acuity. It is unclear whether the olfactory system utilizes a similar organizational scheme to facilitate odor detection. Curiously, olfactory sensory neurons (OSNs) in the mouse nose are differentially stimulated depending on the cell location. We therefore asked whether OSNs in different locations evolve unique structural and/or functional features to optimize odor detection and discrimination. Using immunohistochemistry, computational fluid dynamics modeling, and patch clamp recording, we discovered that OSNs situated in highly stimulated regions have much longer cilia and are more sensitive to odorants than those in weakly stimulated regions. Surprisingly, reduction in neuronal excitability or ablation of the olfactory G protein in OSNs does not alter the cilia length pattern, indicating that neither spontaneous nor odor-evoked activity is required for its establishment. Furthermore, the pattern is evident at birth, maintained into adulthood, and restored following pharmacologically induced degeneration of the olfactory epithelium, suggesting that it is intrinsically programmed. Intriguingly, type III adenylyl cyclase (ACIII), a key protein in olfactory signal transduction and ubiquitous marker for primary cilia, exhibits location-dependent gene expression levels, and genetic ablation of ACIII dramatically alters the cilia pattern. These findings reveal an intrinsically programmed configuration in the nose to ensure high sensitivity to odors.

Anti-inflammatory effects of grape seed procyanidin B2 on a diabetic pancreas.

The prevalence of type 2 diabetes mellitus (T2DM) has increased considerably in recent years, highlighting the importance of developing new therapeutic strategies. Insulin-resistance and gradual dysfunction of pancreatic islets are the mainstay in the progression of T2DM. Therefore, preserving the function of the pancreas may lead to new prospective approaches. Our previous studies suggested that grape seed procyanidin B2 (GSPB2), a natural polyphenol product, exhibited protective effects on diabetic vasculopathy. However, effects of GSPB2 on a diabetic pancreas remain unknown. In this study, we provided strong evidence that GSPB2 exerted protective effects on a diabetic pancreas. GSPB2 attenuated the elevated body weights, food intake and advanced glycation end-product (AGE) levels in db/db mice (p < 0.05), though it had no significant effect on glucose levels. The increased islet sizes, insulin levels, as well as HOMA-IR were also improved by GSPB2 treatment in db/db mice (p < 0.05). Milk fat globule epidermal growth factor-8 (MFG-E8), an estimated target of GSPB2 in our previous studies, was up-regulated in pancreatic tissues whereas GSPB2 treatment down-regulated the protein level (p < 0.05). Since MFG-E8 is highly involved in inflammation, we further investigate pro-inflammatory cytokines interleukin-1ß (IL-1ß) and NLRP3 levels. We found that levels of IL-1ß and NLRP3 increased in a diabetic pancreas while GSPB2 treatment notably attenuated these alterations (p < 0.05). In conclusion, our results suggest that inflammation is involved in the damage of a diabetic pancreas and GSPB2 provides protective effects at least in part through anti-inflammation.

Critical role of prohibitin in endothelial cell apoptosis caused by glycated low-density lipoproteins and protective effects of grape seed procyanidin B2.

Elevated level of glycated low-density lipoproteins (glyLDL) is believed to contribute to endothelial dysfunction, which is involved in the pathogenesis and acceleration of diabetic vascular diseases. Grape seed procyanidin B2 (GSPB2) has been reported to possess protective effects against endothelial dysfunction. However, the underlying mechanism remains unclear. Prohibitin (PHB) is a multifunctional protein implicated in cellular survival and apoptosis. In this study, we showed that glyLDL treatment decreased protein level of PHB, reduced viability, and increased apoptosis in human umbilical vein endothelial cells (HUVEC). PHB overexpression or GSPB2 significantly attenuated apoptosis induced by glyLDL. Moreover, PHB siRNA increased HUVEC apoptosis, along with defective mitochondria and increased levels of cytosol cytochrome c concentration, caspase-3 activity, Bax/Bcl-2 ratio, and phosphorylated Akt, whereas PHB overexpression or GSPB2 restored these changes. Our study identified PHB as an important player responsible for HUVEC apoptosis induced by glyLDL. GSPB2 protected against HUVEC apoptosis at least in part through upregulating PHB. Targeting PHB could be significant in fighting against diabetic vascular complications.

Quantitative proteomics study of protective effects of grape seed procyanidin B2 on diabetic cardiomyopathy in db/db mice.

Diabetic cardiomyopathy is one of the major complications of diabetes mellitus. Oxidative stress appears to play a substantial role in cardiomyopathy. Grape seed procyanidin B2 (GSPB2) has been known as an anti-oxidant in treating diabetes mellitus; however, little is known about its effects and underlying mechanisms on diabetic cardiomyopathy. The present study is to explore the molecular targets of GSPB2 responsible for the anti-oxidative effects in db/db mice by quantitative proteomics. GSPB2 (30 mg/kg body weight/day) were intragastric administrated to db/db mice for 10 weeks. Proteomics of the heart tissue extracts by isobaric tags for relative and absolute quantification analysis was obtained from db/db mice. Our study provides important evidence that GSPB2 protect against cardiomyopathy in diabetes mellitus, which are believed to result from regulating the expression of key proteins involving cardiac fibrosis and proliferation. GSPB2 could be expected to become novel clinical application in fighting against diabetic cardiomyopathy.

Protective effect of proanthocyanidins on anoxia-reoxygenation injury of myocardial cells mediated by the PI3K/Akt/GSK-3ß pathway and mitochondrial ATP-sensitive potassium channel.

The aim of the present study was to examine the protective effect of proanthocyanidins anoxia-reoxygenation injury of myocardial cells and its association with phosphatidylinositol-3-kinase/Akt and glycogen synthase kinase (PI3K/Akt/GSK)-3ß and ATP-sensitive potassium channels. Neonatal rat myocardial cells were cultured and an anoxia-reoxygenation model was established following pretreatment with various drugs. The experiment was divided into five groups according to an experimental scheme. An MTT assay was used to examine the cell survival, and reactive oxygen species (ROS) levels and apoptosis were detected by flow cytometry. Myocardial apoptosis was also examined by terminal deoxynucleotidyl transferase dUTP nick end labeling staining and western blot analysis was employed to detect the expression levels of caspase-3, p-Akt and p-glycogen synthase kinase (GSK)-3ß. The results revealed that myocardial cells in the anoxia-reoxygenation group (A/R) exhibited reduced survival rates, increased ROS levels and enhanced caspase-3 expression, as compared with the control group (CN; P<0.05). However, the increase in p-Akt and p-GSK-3ß expression was not significantly different. In the proanthocyanidin pretreatment group (PC) the myocardial cell survival rate was increased, ROS levels were reduced, caspase-3 expression was decreased and p-Akt and p-GSK-3ß expression levels were significantly increased as compared with the A/R group (P<0.05). Blockade of the PIK3/Akt channel by LY294002 eliminated the protective effects of proanthocyanidins and induced a significant decrease in p-Akt protein and p-GSK-3ß expression levels as compared with the PC group. The inhibitor of mitochondrial ATP-sensitive potassium (mitoKATP) channels, 5-HD, also significantly suppressed the protective effects of proanthocyanidins, but had no evident impact on p-Akt and p-GSK-3ß expression as compared with the PC group. In conclusion, pretreatment with proanthocyanidins had a protective effect on rat myocardial cell anoxia/reoxygenation injury. This effect is associated with the activation of the PI3K/Akt/GSK-3ß signaling pathway and the opening of mitoKATP channels, which may have important roles downstream of PI3K.

PIMT prevents the apoptosis of endothelial cells in response to glycated low density lipoproteins and protective effects of grape seed procyanidin B2.

BACKGROUND: The development of diabetic angiopathy is associated with profound vascular endothelial cells (VEC) dysfunction and apoptosis. Glycated low density lipoproteins (gly-LDL) continuously produced in the setting of diabetic patients play an important role in causing VEC dysfunction and apoptosis. However, the underlying molecular mechanism remains largely elusive. Protein L-isoaspartyl methyltransferase (PIMT) is a widely expressed protein repair enzyme by multiple cell types of arterial wall including VEC. Our previous proteomic studies showed that the expression of PIMT was significantly decreased in the aorta of diabetic rats as compared with control rats and treatment with grape seed procyanidin extracts significantly increased the PIMT expression in diabetic rats. We hypothesized that PIMT plays a critical role in gly-LDL induced VEC apoptosis; grape seed procyanidin B2 (GSPB2) protect against gly-LDL induced VEC apoptosis through PIMT regulation. METHODS AND RESULTS: HUVEC transfected negative control and PIMT siRNA were treated with or without GSPB2 (10 µmol/L) for 48 h. Moreover, HUVEC of PIMT overexpression were stimulated by gly-LDL (50 µg/ml) in the presence or absence of GSPB2 (10 µmol/L) for 48 h. Our results showed that gly-LDL downregulated PIMT expression and PIMT overexpression or GSPB2 significantly attenuated gly-LDL induced VEC apoptosis. PIMT siRNA increased VEC apoptosis with up-regulation of p53, cytochrome c release, caspase-9 and caspase-3 activation. Mechanistically, overexpression of PIMT or GSPB2 increased the phosphorylation of ERK1/2 and GSK3ß in the gly-LDL induced VEC. CONCLUSION: In summary, our study identified PIMT as a key player responsible for gly-LDL induced VEC apoptosis and GSPB2 protect against gly-LDL induced VEC apoptosis by PIMT up-regulation. Targeting PIMT including use of GSPB2 could be turned into clinical application in the fighting against diabetic vascular complications.

[Effect and indications of radiofrequency ablation for the treatment of obstructive sleep apnea-hypopnea syndrome].

OBJECTIVE: To explore the effect and indications of radiofrequency ablation for the treatment of obstructive sleep apnea-hypopnea syndrome. METHOD: Multilevel temperature-controlled radiofrequency therapy of soft palate, uvula, inferior turbinate, and tonsils were applied to 74 adults with obstructive sleep apnea-hypopnea syndrome (OSAHS). There were 16 mild, 23 moderate, and 35 severe cases respectively in this study. Evaluation of mucosal injury and effect of radiofrequency therapy on pain, speech and swallowing were performed early after operation. The volume of targets and length of soft palate and uvula were measured three months after operation. Polysomnography, Epworth Sleepiness Scale and Snoring Scale Score questionnaires were reevaluated six months after operation and compared with the results of pre-operation. Treatment outcome measurements were mainly based on polysomnography. RESULT: By our definition, 5 of 74 patients (6.76%) have been cured and 42 of 74 (56.76%) had improved totally. Mean Apnea-Hypopnea Index (AHI) decreased significantly and mean lowest oxygen saturation value increased significantly postoperatively (P < 0.01). The total effective rate of the patients, whose obstructive sites were all treated by radiofrequency, was remarkably higher than that of the ones, whose obstructive sites were only partly treated by radiofrequency (P < 0.01). The total effective rate of the former was 72.92%. Patients showed a significant decrease in mean score on ESS and SSS postoperatively (P < 0.01). No significant complications were observed in most patients. There were little influence on pain, speech and swallowing. The volume or length of targets decreased obviously three months after operation. CONCLUSION: Radiofrequency can reduce the volume of tissue. The short-term outcomes of radiofrequency were satisfying if obstructive sites had been all treated. This study demonstrates that the characters of radiofrequency are as follows: minimally invasive, safe, efficient, repeatable and multilevel applicable. Temperature-controlled radiofrequency therapy is a safe and effective procedure for hypertrophic infraturbinal when used separately, or as a part of a the combined approach for complex syndromes.