Elderberry diet improved cognitive functions, increased long-term potentiation and BDNF expression, and decreased astrogliosis in rats with diabetic cognitive impairments.

The study aimed to investigate the influence of heme oxygenase-1 (HO-1) on rats with diabetic retinopathy (DR) through the extracellular signal-regulated kinase (ERK) 1/2 signaling pathway. 40 rats were selected and divided into Control group (n=10), diabetes mellitus (DM) group (n=10), cobalt protoporphyrin (CoPP) group (n=10) and zinc protoporphyrin (ZnPP) group (n=10) according to weight. Streptozotocin (STZ) was intraperitoneally injected to establish the DM model in DM, CoPP and ZnPP groups, and CoPP and ZnPP solution was intraperitoneally injected in CoPP and ZnPP groups, respectively. Blood was drawn to determine fasting blood glucose. The changes in the protein and messenger ribonucleic acid (mRNA) levels were evaluated via Western blotting and polymerase chain reaction (qRT-PCR), respectively. Enzyme-linked immunosorbent assay (ELISA) was performed to measure antioxidant capacity and the levels of total reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH) and glutathione peroxidase (GPx). The weight of rats was notably higher in the CoPP group and lower inZnPP group than in the DM group (p<0.05). After induction of DM, compared with those in the DM group, the protein expression levels of Nrf2 and pERK were considerably elevated in the CoPP group (p<0.05) but declined remarkably in the ZnPP group (p<0.05). The levels of total ROS and MDA were notably elevated (p<0.05) in DM and ZnPP groups, with a lowered level of GPx and distinctly elevated levels of MDA and total ROS (p<0.05). Moreover, the mRNA expression level of HO-1 in the retinas of rats was remarkably raised in the DM group and CoPP group (p<0.05), but it declined markedly in the ZnPP group (p<0.05). The red fluorescent aggregation of Nrf2 and pERK proteins was overtly less in the ZnPP group than that in the DM group (p<0.05). HO-1 can affect the level of oxidative stress and intervene in retinopathy in DM rats through the Nrf2/ERK pathway.

Objective To observe the morphological changes of the retina at early stage of streptozotocin(STZ)-induced diabetes mellitus(DM)model,so as to assess the feasibility of using STZ-induced DM as an animal model of early diabetic retinopathy(DR).Methods Sixty-four male SD rats(body weight[180±20]g)were randomly divided into the CON and DM groups(n=32).Rats in DM group received intraperitoneal injection of 1% STZ once and those in the CON group were given same volume of citrate buffer in the same manner.The average body weight and blood glucose were similar in the two groups before intraperitoneal injection.The blood glucose,body weight,and other parameters were determined once a week after intraperitoneal injection.In the 10th week,the morphological changes of the retina were observed by H-E staining, stretched preparation of FITC-dextran perfused retinal blood vessels,and transmission electron microscopy in randomly chosen samples.Results The successful rate of STZ-induced DM model was 100%.The body weight of animals in DM group had no obvious increase after injection,and it even decreased at late stage.The body weight increased gradually in the CON group.The average body weight of DM group([169.9±26.9]g)was significantly lower than that of the CON group([439.2±23.5]g)in the 10th week(P0.001).The average blood glucose of DM group([26.63±4.54]mmol/L)was significantly higher than that of the CON group([6.37±0.49]mmol/L)72hafter injection(P0.001).DM group had a blood glucose16.7mmol/L and CON group had a blood glucose of 5.6-7.4mmol/L throughout the 10weeks.Retina H-E staining showed retinal capillary dilatation and interstitial edema in DM group in the 10th week,and the CON group had no obvious abnormalities.Stretched preparation of FITC-Dextran perfused retinal blood vessels showed vascular tortuosity and caliber irregularity in the DM group,but with no leakage, microvascular tumor or retinal non-perfusion area.The retinal vessel diameter was uniform and the branching was natural and smooth in the CON group.TEM results showed the following retinal ultrastructural changes in the DM group:thicker capillary basement membrane,digitation of capillary endothelial cells,mitochondrial swelling,cristae disruption,and vacuolar degeneration in capillary endothelial cells,bipolar cells and ganglion cells,mitochondrial swelling and cristae disruption in pericytes,decreased membranous disc (photoreceptor cell’s outer segment),and widened gap between membranous discs.Conclusion The retina morphological changes of early stage background diabetic retinopathy(BDR)can be found in the 10th week after STZ injection in rats,and STZ-induced DR model can be used as an early stage DR model;besides,the method is simple,economical,quick,with good reproducibility and high successful rate.

The efficacy of statins is related to the ‘common soil’ hypothesis, which proposes oxidative stress and inflammation as main pathophysiologic processes in the disease group of diabetes and endothelial dysfunction. This study evaluated the recovery of erectile function after administration of chronic statin alone in streptozotocin (STZ)-induced diabetes mellitus (DM) rats, focusing on the anti-oxidative effects and consequentially recuperated endothelial function. A total of 45 male Sprague-Dawley rats (8 weeks old) were divided into three groups (n = 15 each): an age-matched normal control group (Control group), an uncontrolled DM group (DM group), and a statin-treated group (Statin group). The rats in the DM and Statin group received an injection of STZ (60 mg/kg). Beginning 10 weeks after the establishment of DM, the Statin group received daily treatment with atorvastatin (10 mg/kg) via oral gavage for four weeks. After 14 weeks, the results of the experiment were evaluated. The ratios of intracavernosal pressure (ICP) to mean arterial pressure (MAP) were recorded with cavernosometry (20 Hz, 3 V, 0.2 msec for 30 seconds) before and after the intravenous administration of udenafil (1 mg/kg). Expression of alpha-smooth muscle actin (α-SMA) was evaluated using cavernosal tissue. In addition, changes in RhoA translocation ratio and myosin phosphatase target subunit 1 (MYPT1) phosphorylation were evaluated with western blot. Superoxide dismutase (SOD) and malondialdehyde (MDA) levels were also analyzed as measurements of oxidative stress levels. The ICP/MAP and area under the curve (AUC)/MAP ratios of the Statin group were obviously superior to the DM group, but were not comparable to the Control group (P<0.001). The level of oxidative stress, namely SOD activity, was also significantly lower in the Statin group than in the DM group (P = 0.015), and was comparable to the Control group. In contrast, MDA levels were not considerably different among the groups (P = 0.217). The RhoA translocation ratio was not significantly different among the groups (P = 0.668), whereas MYPT1 phosphorylation in the Statin group was significantly lower than in the DM group (P = 0.030), and similar to the Control group. Expression of α-SMA in the Statin group was higher than in the DM group (P<0.001), and comparable to the Control group. Chronic statin treatment alone showed anti-oxidative effects and helped to restore the erectile mechanism, but did not lead to the full recovery of erectile function in STZ-induced DM rats. Therefore, combination therapy rather than a single agent should be the preferred treatment strategy for DM-associated erectile dysfunction, especially in the setting of severe diabetes.

Effects of comorbidities on the outcomes of manipulation under anesthesia for primary stiff shoulder.

Though diabetes mellitus (DM) is one of the known causes of osteoporosis, it is also realized that ketogenic diet (KD), an effective regimen for epilepsy, impairs bone microstructures. However, the similarities and differences of effects between these two factors are still unknown. The purpose of this study is to identify different effects between hyperglycemia and hyperketonemia, which are manifestations of DM and KD, on bone in rats. Thirty male Sprague-Dawley rats were randomly divided into three groups: the sham, DM, and KD groups. Hyperglycemia was achieved by intravenous injection of streptozotocin in DM group, while hyperketonemia was induced by application of ketogenic diet (carbohydrates-to-fat as 1:3) in KD group. The body weight, blood ketone body, and blood glucose were recorded, and the bone turnover markers, bone length, bone microstructures, bone biomechanics and histomorphology were measured after 12 weeks intervention. Compared with the control and KD groups, a significant body weight loss was found in the DM group, and the bone lengths of tibia and femur of the group were shortened. The blood glucose and blood ketone were noticeably increased in the DM and KD rats, respectively. Microstructures and properties of cancellous bone were significantly deteriorated in both the DM and KD groups compared with the sham group, as the bone volumes were decreased and the bone trabecula structures were disturbed. Meanwhile, the thickness and strength of cortical bone was reduced more in the DM group than those in the sham and KD groups. The HE staining showed that bone trabecula was significantly decreased in both the DM and KD groups, and more adipose tissue was observed in the KD rats. The activity of osteoblasts was decreased more in both the KD and DM groups than that in the sham group, while the activity of osteoclasts of the two groups was remarkably increased. The present study indicates that both hyperglycemia and hyperketonemia have adverse effects on bone. Therefore, it is worth paying more attention to the bone status of patients with hyperglycemia and hyperketonemia in clinic.

Effect of Diabetes Mellitus on Muscle Size and Strength in Patients Receiving Dialysis Therapy

Many studies have demonstrated that diabetes mellitus (DM) is associated with cardiac dysfunction. Exercise training is beneficial for the treatment of chronic heart failure. PURPOSE: the aim of the present study was to evaluated effects of endurance exercise training (EX) on cardiac function quantification by echocardiography in DM rats. We also evaluated the levels of connexin 43 of the heart. METHODS: the Sprague-Dawley (SD) male rats were divided into 3 groups as two experimental groups and one control group. The first groups as a control group received adjuvant. The second and third groups were the DM groups, received 150 mg/kg STZ by intraperitoneal injection. After one week, EX was carried out in the third group (DM-EX group) for four weeks. The cardiac functions of those animals were evaluated by echocardiography method. The animals were sacrificed; the connexin 43 of heart was evaluated by western blot. The tissues of heart were stained by H&E or Masson’s trichrome stain. The morphology and fibrosis of cardiac cells were analyzed. RESULTS: the results show that the distances of interventricular septal and left ventricular (LV) posterior wall were shorter in DM and DM-EX groups than in control group (P<0.05). The LV diameter was longer in DM-EX group than in control group (P<0.05). The weights of rat’s body, heart and LV mass were lower in DM and DM-EX groups than in control group (P<0.05). The values of heart and LV mass weighted by body mass were higher in DM group than in control group (P<0.05). The heart connexin 43 was lowest in DM group than in control and DM-EX group (P<0.05). The length of cardiomyocyte diameter was longer in DM and DM-EX group than in control group (P<0.05). The percent of fibrotic area in heart tissue was highest in DM group than in others (P<0.05). The fibrotic area of heart was smaller in DM-EX group than in DM group (P<0.05). CONCLUSIONS: those results suggested that EX may decrease cardiac dysfunction from DM damage through connexin 43 increasing and lower myocardial fibrosis.

Background:Rheumatoid arthritis (RA) patients can experience various comorbidities1. The incidence of diabetes mellitus (DM) is reported higher in patients with RA2 and comorbid DM is likely to affect treatment outcomes3 and then healthcare resource uses, however, no previous study has not focused on it.Objectives:To evaluate medical costs and resource use in patients starting treatment for RA with and without DM using a large claims database in Japan.Methods:We used a large Japanese administrative claims database constructed by the Japan Medical Data Center (JMDC)4. Patients with the International Classification of Diseases 10th revision (ICD-10) codes for RA who started medication with disease-modifying antirheumatic drugs (DMARDs) after 6 months without them in the period from 1/1/2012 to 12/31/2017 and who were observable for 12 months as a follow-up period were enrolled. These patients were categorized as DM or non-DM group with ICD-10 codes for DM plus use of antidiabetic drugs in 6 months before starting DMARDs (baseline period). To adjust baseline characteristics between the 2 groups, they were matched by sex, age, Charlson Comorbidity Index (CCI) except for DM, months from the first RA codes to starting DMARDs, and medications. The primary endpoint was mean medical cost per patient in the 12-month follow-up period. Costs in JPY were converted into EUR (1 EUR = 125 JPY in 2020). Costs for drugs, treatments, and materials and their subcategories were evaluated both with and without DM-specific costs. The secondary endpoints were the proportions of patients using the subcategories of each resource.Results:Patients of 161 for the DM group and 2,974 for the non-DM group were eligible, and 109 patients were matched from each group. The medians of age and CCI were 59 years and 2.0 in both groups and no significant difference was observed in all baseline characteristics used for matching between the groups. Total mean costs were significantly higher in the DM group (DM, 5,331 EUR, non-DM 3,200 EUR; P&lt; 0.05). After excluding DM-specific costs, drug costs were significantly higher in the DM group than in the non-DM group (DM 1,883 EUR, non-DM 896 EUR; P &lt; 0.05), especially costs for biological DMARDs (DM 1,156 EUR, non-DM 292 EUR; P &lt; 0.05), mainly because a higher proportion of patients used these drugs in the DM group (Table 1). Treatment costs (DM 2,380 EUR, non-DM 2,133 EUR) and material costs (DM 74 EUR, non-DM 149 EUR) were not different between the groups, but only costs for examinations were significantly higher in the DM group (DM 970 EUR, non-DM 779 EUR; P &lt; 0.05).Table 1.Number and proportion of patients who used drugsType of drugDrug use, n (%)DM (N = 109)Non-DM (N = 109)P-valuecsDMARDsTotal109 (100.0)109 (100.0)1.000Methotrexate101 (92.7)102 (93.6)1.000Others46 (42.2)51 (46.8)0.583bDMARDsTotal16 (14.7)6 (5.5)0.041TNFi11 (10.1)4 (3.7)0.118IL6i6 (5.5)2 (1.8)0.219T-cell4 (3.7)0 (0.0)0.125tsDMARDs0 (0.0)0 (0.0)1.000CSs65 (59.6)62 (56.9)0.711AnalgesicsTotal103 (94.5)96 (88.1)0.167Acetaminophen24 (22.0)23 (21.1)1.000Acetaminophen /Opioids10 (9.2)6 (5.5)0.454NSAIDs102 (93.6)93 (85.3)0.093Opioids0 (0.0)4 (3.7)0.125Others25 (22.9)17 (15.6)0.185bDMARDs=biological disease-modifying antirheumatic drugs; CSs=corticosteroids; csDMARDs=conventional synthetic disease-modifying antirheumatic drugs; DM=diabetes mellitus; IL6i=interleukin-6 inhibitor; NSAID=non-steroidal anti-inflammatory drug; T-cell=selective T-cell co-stimulation modulator; TNFi=tumor necrosis factor α inhibitor; tsDMARDs=targeted synthetic disease-modifying antirheumatic drugs; P-values were calculated using McNemar testConclusion:Medical costs for RA were higher in the DM group than in the non-DM group because of more prevalent use of biological DMARDs in the DM group.

To explore the influence of micro ribonucleic acid (miR)-135a on the renal fibrosis in rats with diabetic kidney disease (DKD) through the Notch signaling pathway. A total of 30 male Wistar rats weighing 200-220 g were selected and randomly divided into Control group (n=10), diabetes mellitus (DM) group (n=10), and miR-13a inhibitor group (n=10). Streptozotocin (STZ) was intraperitoneally injected daily to establish the DM model in rats of both DM group and miR-135a group, while normal saline was given daily through intraperitoneal injection in rats of Control group. After 4 weeks, the rats in miR-135a inhibitor group were intraperitoneally injected with miR-135a inhibitor, and those in Control and DM groups were administrated with an equal amount of normal saline. Changes in the blood glucose (BG), glycated hemoglobin (GHb), serum creatinine (Scr), triglyceride (TG), and total cholesterol (TC) of rats were evaluated, and the pathological changes in the renal tissues of DM rats were observed via hematoxylin-eosin (HE) staining. Sirius red staining was performed to observe the changes in collagen fibers in the kidney of all groups of rats. The expressions of Notch and Hes1 in the renal tissues of rats in each group were detected using immunohistochemistry. Immunofluorescence assay was employed to detect the positive expression of Notch in the renal tissues of rats. The mRNA expressions of Notch and miR-135a were detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Finally, Western blotting was conducted to detect the protein expressions of Notch, Notch intracellular domain (NICD) and Hes1. Compared with Control group, rats in DM group had substantially raised levels of BG, GHb, Scr, TG, and TC (p<0.05). HE staining showed that the rats in Control group had renal tubular cells with normal morphology and well-defined structure, while those in DM group exhibited evident cavitation in the renal tubular epithelium. Sirius red staining results manifested that the red collagen fibers were evenly distributed with light staining in the glomeruli and renal tubules of rats in Control group. In contrast, the collagen fibers of the glomeruli and renal tubules of rats in DM group exhibited deep and evident red staining. Moreover, compared with DM group, rats in miR-135a inhibitor group had notably faded red staining in the glomeruli and renal tubules of rats, evenly distributed collagen and remarkably decreased fibrotic nodules. According to immunohistochemistry detection results, the protein levels of Notch and Hes1 in the renal tubulointerstitial cells and renal tubular epithelial cells of rats in DM group were markedly higher than those in Control group. Compared with those in DM group, their protein levels were remarkably lowered in miR-135a inhibitor group (p<0.05). Immunofluorescence assay results revealed that the protein level of Notch in the renal tissues of rats in DM group was considerably higher than that in Control group (p<0.05), while its protein level in miR-135a inhibitor group was significantly lower than that in DM group. According to qRT-PCR results, compared with those in Control group, mRNA expressions of Notch mRNA and miR-135a in the rat kidney tissues were substantially raised in DM group (p<0.05), and they were notably lowered in miR-13a inhibitor group compared with those in DM group (p<0.05). Finally, Western blotting results manifested that the protein levels of Notch, NIC, and Hes1 in the renal tissues of rats in DM group were considerably higher than those in Control group (p<0.05), and that their protein expression levels in miR-135a inhibitor group were markedly lower than those in DM group (p<0.05). Inhibition of miR-135a can reduce the renal fibrosis in DKD rats through the Notch pathway.

Investigation of Zinc on hemorheological parameters in a rat model of diabetes

Objective To investigate the effects of regular insulin (RI)on duodenal smooth muscle in diabetic mice. Methods Diabetes mellitus (DM) model was established by intraperitoneal injection of 150 mg/kg streptozotocin (STZ) in male BALB/c mice. The model mice were divided into DM group and DM treated with RI group with 6 each. Meanwhile, 6 normal mice were served as controls. The mice in treatment group were intraperitoneally injected with 40 U/kg of RI daily.Whereas the mice in DM and control groups were intraperitoneally injected with phosphate buffer solution (pH = 7. 40). After 6 weeks, the small intestinal transit rate of mice was determined by lavage of Indian ink. Interstitial cells of cajal (ICC) in duodenal myenteric plexus were counted using immunohistochemical staining. Slow waves of duodenal smooth muscle cells were recorded with intracellular recordings. Data were analysed by SPSS 17.0 software, and comparisons among three groups were done using LSD test. Results After intervention for 6 months, the clinical presentations,such as more water and food intake and polyuria, were improved in treatment group. The body weight was increased in treatment group [(23.33±3.13) g] compared with DM group [(15.42±1.40) g,P＜0.01] ,but dereased compared with control group [(26.78 ± 2.09) g, P＜0.05]. The level of blood glucose in DM group was significantly higher than that in control and treatment groups(P＜0.01). Small intestine transmission rate was significantly reduced in DM group than that in control and treatment groups (P＜0.01), but it was slower in treatment group than that in control group (P＜ 0. 01 ). Immunohistochemical study showed that the number of c-kit positive cells reduced obviously in DM group than that in control group and treatment group (P＜0.05), whereas it was lower in treatment group than that in control group (P ＜ 0.05). The slow wave frequency and amplitude of duodenal smooth muscle cells in DM group were reduced when compared with control and treatment groups (P＜0.01) and both were lower in treatment group than that in control group (P＜0. 01 ). Conclusion The findings indicate that DM mice have gastrointestinal dysmotility and exogenous insulin may improve small intestinal dysmotility in DM mice. Key words: Diabetes mellitus; Insulin; Duodenum; Myocytes, smooth muscle

The present study investigated the association between the RAGE G82S polymorphism, the plasma levels of sRAGE and chronic periodontitis in subjects with and without diabetes mellitus (DM). A total of 230 patients with DM and 264 non-DM participants were recruited for this study. Genotyping of the RAGE G82S polymorphism was accomplished using polymerase chain reaction-restriction fragment length polymorphism, and associations were analyzed with the chi-squared test and logistic regression analysis. In the non-DM group, the chi-squared test showed that the frequency distributions of the G82S polymorphism were significantly different between chronic periodontitis and non-chronic periodontitis subjects (χ(2) = 8.39, p = 0.02). A multivariate logistic regression model showed that the (G82S + S82S) genotypes were associated with a significantly increased risk of chronic periodontitis development compared to the G82G genotype (adjusted odds ratio = 2.06, 95% confidence interval: 1.08-4.07). In the DM group, there was no association between the G82S polymorphism and chronic periodontitis development when a multivariate logistic regression was performed. Plasma levels of sRAGE were significantly higher in subjects with the G82G genotype compared to those with the (G82S + S82S) genotypes in both the non-DM (856.6 ± 332.0 vs. 720.4 ± 311.4 pg/mL, p = 0.003) and DM groups (915.3 ± 497.1 vs. 603.5 ± 298.3 pg/mL, p < 0.0001). However, there was no difference in plasma sRAGE levels between chronic periodontitis and non-chronic periodontitis subjects in both the DM and non-DM groups. Moreover, when the subjects were further sub-divided by the G82S polymorphism, the difference in plasma levels of sRAGE between chronic periodontitis and non-chronic periodontitis subjects in the DM and non-DM groups remained statistically insignificant. The present study revealed that the RAGE G82S polymorphism was associated with chronic periodontitis in the non-DM group but not in the DM group. Our results also showed that the plasma levels of sRAGE were significantly higher in subjects with the RAGE G82G genotype, and this correlation was not affected by the presence of chronic periodontitis in the DM and non-DM groups.

Objective To investigate the effect of valsartan on the expressions of toll-like recpter 4 (TLR 4) and myeloid differentiation factor 88 (Myd88) in the arteria of diabetic rats. Methods Thirty-one Sprague-Dawley rats were randomly divided into normal control group (n = 7), diabetes mellitus group (n=12) and valsartan group (n=12). Diabetic rats induced by a single injection of streptozotocin were treated with valsartan 20mg·kg-1·d-1 intragastrically.The blood glucose and body weight were monitored periodically. The expressions of TLR4, Myd88 mRNA and protein in the arteria of the rats at 12th week were analyzed by RT-PCR, immunohistochemistry and Western blotting. Results The body weight of rats in diabetes mellitus and valsartan groups decreased significantly compared with that in normal control group (P 0.05). The expressions of TLR4 and MyD88 mRNA and protein in diabetes mellitus group were significantly increased as compared with normal control group(both P<0.01), but decreased after valsartan treatment (P<0.01). Conclusions Valsartan significantly reduces the expressions of TLR4 and MyD88 in the arteria of diabetic rats, suggesting that may be useful for preventing or treating early inflammation in the arteria of diabetic rats. Key words: Diabetes mellitus,experimental; Rats,Sprague-Dawley; Arteria; Toll-like receptor 4; Myeloid differentiation factor 88; Valsartan

Response

Although skeletal muscle abnormalities caused by diabetes mellitus (DM) suggest that peripheral muscle impairment may have a greater effect on exercise tolerance in patients with DM, the magnitude of this effect on reduced exercise capacity remains unclear. As such, this study aimed to compare the strength of the association between lower-extremity muscle strength and exercise capacity in patients diagnosed with cardiovascular disease (CVD) with and without DM. This retrospective cross-sectional study included data from 262 patients divided into two groups: patients with DM (DM group; n =106); and without DM (non-DM group; n =156). Peak oxygen uptake (VO 2peak ) and isometric knee extensor strength (IKES) were measured. Correlations between VO 2peak and IKES were analyzed using Pearson's correlation test in the DM and non-DM groups. Linear regression analyses were performed with VO 2peak as the dependent variable, and IKES, confounders, and the interaction term DM× IKES as the independent variables. Separate linear regression analyses were performed for the DM and non-DM groups. The correlation coefficient between VO 2peak and IKES was 0.58 in the DM group and 0.26 in the non-DM group. The interaction term DM× IKES had a significant effect on VO 2peak. The IKES was significantly associated with VO 2peak in the DM group ( β =0.83, P < .001), but not in the non-DM group ( β =0.01, P =.96). A specific association between lower-extremity muscle strength and VO 2peak was observed in patients with both CVD and DM.