PDE-5-Inhibited BMSCs Alleviate High Glucose-Induced Myocardial Fibrosis and Cardiomyocyte Apoptosis by Activating the cGMP/PKG Pathway.

BACKGROUND: Phosphodiesterase-5 (PDE-5) inhibitors have been found to play an important cardio-protective role. This study aimed to clarify the inhibitory effects of PDE-5-silenced bone marrow mesenchymal stem cells (BMSCs) on high glucose-induced myocardial fibrosis and cardiomyocyte apoptosis. METHODS: Cardiomyocytes and fibroblasts of neonatal rats were treated with high glucose (HG), and co-cultured with PDE-5-overexpressed or -knocked down BMSCs. The viability and apoptosis as well as the levels of cytokines, Cardiac troponin I and Vimentin of cardiomyocytes and fibroblasts were studied. The expressions of PDE-5, cyclic guanosine monophosphate (cGMP) and protein kinase G (PKG), in both cells were evaluated. RESULTS: BMSCs that silenced PDE-5 facilitated the viability of cardiomyocytes, decreased the viability of fibroblasts, and inhibited the apoptosis of cardiomyocytes and fibroblasts. The contents of collagen-I, collagen-III, tissue inhibitor of metalloproteinase (TIMP)-1 and Dermin in fibroblasts were decreased by the PDE-5 inhibitor, but the levels of matrix metalloproteinase (MMP)-1 in fibroblasts and troponin-I in cardiomyocytes were increased by the PDE-5 inhibitor. PDE-5 inhibitor also suppressed the expression of PDE-5 but up-regulated cGMP and PKG expression in cardiomyocytes and fibroblasts. CONCLUSIONS: PDE-5-inhibited BMSCs can decrease HG-induced myocardial fibrosis and cardiomyocyte apoptosis by activating the cGMP/PKG pathway, and may play a role in the prevention and treatment of diabetic cardiomyopathy.

Urinary N-acetyl-ß-d-glucosaminidase-creatine ratio is a valuable predictor for advanced diabetic kidney disease.

BACKGROUND: Many biomarkers show high diagnostic values for diabetic kidney disease (DKD), but fewer studies focus on the predictive assessment of DKD progression by blood and urinary biomarkers. AIM: This study aims to find powerful risk predictors and identifying biomarkers in blood and urine for DKD progression. METHODS: A total of 117 patients with type 2 DKD including early and advanced stages and their laboratory parameters were statistically assessed. A receiver operating characteristic (ROC) curve analysis was performed to evaluate the significance of discriminating between early and advanced DKD, and the predictive power for advanced DKD was analyzed by regression analysis and trisector grouping. RESULTS: N-acetyl-ß-d-glucosaminidase-creatine (NAG/CR) level in advanced DKD was statistically higher than that in early DKD (p < 0.05), and there was a higher incidence of advanced DKD (72% vs. 56%) and high odds ratio (OR: 3.917, 95% CI: 1.579-10.011) of NAG/CR with ≥2.79 U/mmol compared with <2.79 U/mmol (p < 0.05). NAG/CR ratio also showed a higher area under the ROC curve of 0.727 (95% CI: 0.616-0.828, p = 0.010) with a high sensitivity (0.75) and a moderate specificity (0.66) when 1.93 U/mmol was set as the optimal cutoff value. The adjusted-multivariable analysis revealed that NAG/CR had an OR of 1.021 (95% CI: 1.024-1.038) and 2.223 (95% CI: 1.231-4.463) based on a continuous and categorical variable, respectively, for risk of advanced DKD. Moreover, the prevalence of advanced DKD exhibited an increasing tendency by an increment of the trisector of NAG/CR. CONCLUSIONS: This study suggests that NAG/CR ratio is an independent predictor for advanced DKD, and it also can be used as a powerful identifying marker between early and advanced DKD.

Clinical and predictive significance of Plasma Fibrinogen Concentrations combined Monocyte-lymphocyte ratio in patients with Diabetic Retinopathy.

Diabetic retinopathy (DR) is one of the most common causes of blindness and visual impairment. Therefore, early prediction of its occurrence and progression is important. This study aimed to assess the clinical and predictive significance of plasma fibrinogen concentrations combined monocyte-lymphocyte ratio (FC-MLR) in patients with DR. A total of 307 patients with type 2 diabetes (T2D) were enrolled. Plasma fibrinogen concentrations and peripheral white blood cells were measured, and MLR was calculated, and the associations of FC-MLR with DR and severity of disease were assessed. Regression analysis and receiver operating characteristic (ROC) curves were performed to evaluate the risk factors and predictive power of FC-MLR for DR and severity of disease, respectively. DR patients showed higher fibrinogen concentrations and a higher MLR than did T2D patients without complications (P<0.01); Moreover, DR patients in proliferative stage also showed higher fibrinogen concentrations and a higher MLR than did those in non-proliferative stage (P<0.01). FC-MLR was closely associated with occurrence and severity of DR (P<0.01), and was an independent risk factor for them (OR=6.123, 95%CI: 3.122-17.102; and 7.932, 95%CI: 4.315-16.671, respectively; P<0.001). The predictive sensitivity and specificity for DR and severity of disease were 0.86 and 0.68, and 0.85 and 0.73, respectively. The study suggests that FC-MLR may be used as a predictor for the risk and progression of diabetic retinopathy.

Monocyte-lymphocyte ratio is a valuable predictor for diabetic nephropathy in patients with type 2 diabetes.

Diabetic nephropathy (DN) is serious threat to human health. Therefore, early prediction of its occurrence is important. This study aimed to assess the predictive significance of monocyte-lymphocyte ratio (MLR) for DN.A total of 301 patients with type 2 diabetes (T2D), including 212 T2D patients without diabetic-related complications and 99 DN patients, were enrolled. Peripheral white blood cells were measured before treatment to calculate MLR, and the risk factors and predictive significance for T2D and DN were assessed.T2D patients without diabetic-related complications had higher MLR than control patients (P < .01). However, MLR was significantly higher in DN patients than in T2D patients without diabetic-related complications (P < .001). According to MLR quartiles, higher MLR in DN patients was correlated with higher serum creatinine, estimated glomerular filtration rate, and urinary albumin excretion (UAE) levels (P < .01 or P < .001). Furthermore, MLR was positively correlated with UAE level (R = 0.5973; P < .01) and an independent predictor for DN (odds ratio: 7.667; 95% confidence interval [CI]: 3.689-21.312; P < .001). The area under the receiver-operating characteristic (ROC) curve for MLR was 0.874 (95%CI: 0.830-0.918, P < .001). When the optimal cutoff value was 0.23, the sensitivity and specificity of MLR for DN prediction were 0.85 and 0.74, respectively.The present findings suggest that MLR is a powerful independent predictor for DN.

Combining infiltration holes and level ditches to enhance the soil water and nutrient pools for semi-arid slope shrubland revegetation.

Active vegetation restoration has been proposed as an effective approach for restoring degraded ecosystems. Soil water and nutrient deficits hinder slope revegetation in arid and semi-arid areas. However, few studies have discussed rainfall runoff utilization and soil nutrient conservation within the context of slope vegetation restoration. In this study, the effects of combining infiltration holes and level ditches on the soil water storage, organic matter, and total nitrogen were analyzed on the slopes of shrubland and bare land. The results showed that the combined measures significantly increased the average soil water content above the 100 cm soil layer and mitigated soil desiccation below 220 cm in the shrubland. Meanwhile, the combined measures obviously increased the soil organic matter and total nitrogen above the 60 and 40 cm soil layers in bare land and shrubland, respectively. Overall, combining infiltration holes and landscape engineering measures is an effective approach for enhancing the soil water and nutrient pools of slopes. Our findings provide an effective engineering measure to combat soil water and nutrient deficits for slope vegetation restoration in arid and semi-arid areas.

Enkephalinergic Circuit Involved in Nociceptive Modulation in the Spinal Dorsal Horn.

Enkephalin (ENK) has been implicated in pain modulation within the spinal dorsal horn (SDH). Revealing the mechanisms underlying ENK analgesia entails the anatomical and functional knowledge of spinal ENK-ergic circuits. Herein, we combined morphological and electrophysiological studies to unravel local ENK-ergic circuitry within the SDH. First, the distribution pattern of spinal ENK-ergic neurons was observed in adult preproenkephalin (PPE)-GFP knock-in mice. Next, the retrograde tracer tetramethylrhodamine (TMR) or horseradish peroxidase (HRP) was injected into the parabrachial nucleus (PBN) in PPE-GFP mice. Immunofluorescent staining showed I-isolectin B4 (IB4) labeled non-peptidergic afferents were in close apposition to TMR-labeled PBN-projecting neurons within lamina I as well as PPE-immunoreactivity (-ir) neurons within lamina II. Some TMR-labeled neurons were simultaneously in close association with both IB4 and PPE-ir terminals. Synaptic connections of these components were further confirmed by electron microscopy. Finally, TMR was injected into the PBN in adult C57BL/6 mice. Whole-cell patch recordings showed that δ-opioid receptor (DOR) agonist, [D-Pen2,5]-enkephalin (DPDPE, 1 µM), significantly reduced the frequency of miniature excitatory postsynaptic current (mEPSC) and decreased the activity of TMR-labeled neurons. In conclusion, spinal ENKergic neurons receive direct excitatory inputs from primary afferents, which might be directly recruited to release ENK under the condition of noxious stimuli; ENK could inhibit the glutamatergic transmission towards projecting neurons via presynaptic and postsynaptic DORs. These morphological and functional evidence may explain the mechanisms underlying the analgesic effects exerted by ENK within the SDH.

Expression of NR1 and apoptosis levels in the hippocampal cells of mice treated with MK­801.

The aim of the present study was to investigate the characteristics of N­methyl­D­aspartate receptor R1 (NR1) expression and apoptosis in the nerve cells of the hippocampus in schizophrenia­like mice. C57BL/6 mice were randomly allocated to the following groups: i) Blank group; ii) MK­801 group; iii) MK­801+NMDA group, according to body weight. The NMDAR antagonist, MK­801 (0.6 mg/kg/d) was intraperitoneally injected daily for 14 days to induce a schizophrenia­like phenotype mouse model, and the effect of the NMDA injection via the lateral ventricle was observed. The results demonstrated that the number of NR1 positive cells in the MK­801 group increased in the CA1 and DG regions, indicating that NMDA may reverse this change. The level of damage decreased in the MK­801 treated group when compared with the blank group in the CA3 region. The protein expression of NR1 increased however, at the mRNA expression level, NR1 was lower in the MK­801 treated group when compared to the blank group; NMDA also reversed this change. In addition, early and total apoptosis detected in the hippocampal nerve cells was significantly increased in the MK­801 group when compared with the blank group, which was reversible following treatment with NMDA. These results indicated that NMDA may regulate the expression of NR1 and suppress apoptosis in hippocampal nerve cells in schizophrenia­like mice. Thus, NR1 may be a promising therapeutic target for the treatment of schizophrenia.

[Characteristics and Mechanism of Copper Adsorption from Aqueous Solutions on Biochar Produced from Sawdust and Apple Branch].

In order to utilize the waste biomass resources efficiently, two raw materials of sawdust and apple branch were selected to produce biochars at 450℃ by oxygen-limited pyrolysis, and the adsorptions of Cu2+ in aqueous solutions were evaluated. The effects of initial Cu2+ concentration and contact time in batch sorption experiments were investigated by the four kinds of isothermal adsorption models (Langmuir, Freundlich, Temkim, D-R model) and the four kinds of adsorption kinetics models (Pseudo first-order, Pseudo second-order, Elovich model, Intraparticle diffusion model). The influencing factors of adsorption characteristics about sawdust biochar (JB) and apple branch biochar (PB) were discussed by characterizing their elemental composition, functional groups, surface area, surface morphology and scanning electron microscope. The results showed that:1The adsorption equilibrium of PB and JB was reached in three hours and six hours, respectively, and the maximum adsorption capacity of PB and JB reached 15.85 mg·g-1 and 17.44 mg·g-1, respectively. Compared with other studies, these biochars showed higher Cu2+ adsorption performance. 2The adsorption kinetics was best fitted by the pseudo-second order model, while the isothermal adsorption was best described by Langmuir isotherms. This indicated that the beneficial adsorption process via monolayer was affected by intraparticle diffusion, surface adsorption and liquid film diffusion. 3The adsorption mechanism for Cu2+ is a complex interaction of physical and chemical factors. Electrostatic interaction of physical adsorption is mainly adsorption mechanism, Chemical adsorption mechanism includes generally ligand exchange (phenolic hydroxyl)/ion exchange (H+) and cationic-π. It was demonstrated that JB and PB could be considered as promising materials to immobilize heavy metals in contaminated water or soil.

Directed differentiation of postnatal hippocampal neural stem cells generates nuclear receptor related­1 protein­ and tyrosine hydroxylase­expressing cells.

Parkinson's disease (PD) is a severe neurodegenerative disorder. Although the detailed underlying molecular mechanism remains to be elucidated, the major pathological feature of PD is the loss of dopaminergic (DA) neurons of the substantia nigra. The use of donor stem cells to replace DA neurons may be a key breakthrough in the treatment of PD. In the present study, the growth kinetics of hippocampal neural stem cells (Hip­NSCs) isolated from postnatal mice and cultured in vitro were observed, specifically the generation of cells expressing DA neuronal markers nuclear receptor related­1 protein (Nurr1) and tyrosine hydroxylase (TH). It was revealed that Hip­NSCs differentiated primarily into astrocytes when cultured in serum­containing medium. However, in low serum conditions, the number of ßIII tubulin­positive neurons increased markedly. The proportion of Nurr1­positive cells and TH­positive neurons, significantly increased with increasing duration of directed differentiation of Hip­NSCs (P=0.0187 and 0.0254, respectively). The results of the present study reveal that Hip­NSCs may be induced to differentiate in vitro into neurons expressing Nurr1 and TH, known to be critical regulators of DA neuronal fate. Additionally, their expression may be necessary to facilitate neuronal maturation in vitro. These data suggest that Hip­NSCs may serve as a source of DA neurons for cell therapy in patients diagnosed with PD.

TLQP-21 protects human umbilical vein endothelial cells against high-glucose-induced apoptosis by increasing G6PD expression.

Hyperglycemia causes oxidative stress that could damage vascular endothelial cells, leading to cardiovascular complications. The Vgf gene was identified as a nerve growth factor-responsive gene, and its protein product, VGF, is characterized by the presence of partially cleaved products. One of the VGF-derived peptides is TLQP-21, which is composed of 21 amino acids (residues 556-576). Past studies have reported that TLQP-21 could stimulate insulin secretion in pancreatic cells and protect these cells from apoptosis, which suggests that TLQP-21 has a potential function in diabetes therapy. Here, we explore the protective role of TLQP-21 against the high glucose-mediated injury of vascular endothelial cells. Using human umbilical vascular endothelial cells (HUVECs), we demonstrated that TLQP-21 (10 or 50 nM) dose-dependently prevented apoptosis under high-glucose (30 mmol/L) conditions (the normal glucose concentration is 5.6 mmol/L). TLQP-21 enhanced the expression of NAPDH, resulting in upregulation of glutathione (GSH) and a reduction in the levels of reactive oxygen species (ROS). TLQP-21 also upregulated the expression of glucose-6-phosphate dehydrogenase (G6PD), which is known as the main source of NADPH. Knockdown of G6PD almost completely blocked the increase of NADPH induced by TLQP-21, indicating that TLQP-21 functions mainly through G6PD to promote NADPH generation. In conclusion, TLQP-21 could increase G6PD expression, which in turn may increase the synthesis of NADPH and GSH, thereby partially restoring the redox status of vascular endothelial cells under high glucose injury. We propose that TLQP-21 is a promising drug for diabetes therapy.

No association between IRS­1 promoter methylation and type 2 diabetes.

As a candidate gene for type 2 diabetes (T2D), insulin receptor substrate-1 (IRS­1) gene variations were found to be associated with the risk of T2D. The aim of our study was to investigate the contribution of promoter DNA methylation of the IRS­1 gene to the risk of T2D. Using bisulphite pyrosequencing technology, the DNA methylation levels of 3 CpG dinucleotides within the IRS­1 gene promoter were measured in 48 T2D patients and 48 age­ and gender­matched healthy controls. Our results indicated that there was no significant association between the methylation of the IRS­1 gene promoter and the risk of T2D (P>0.1). A breakdown analysis by gender revealed that IRS­1 promoter methylation was not associated with an increased risk of T2D for either gender (P>0.1), although there were significantly lower methy-lation levels of CpG1 (P=0.002) and CpG2 (P=0.043) within the IRS­1 gene promoter in males than in females.