Messenger Ribonucleic Acid Levels Research Articles

Functional adenosine triphosphate-sensitive potassium channel is required in high-carbohydrate diet-induced increase in β-cell mass.

Aims/IntroductionA high‐carbohydrate diet is known to increase insulin secretion and induce obesity. However, whether or not a high‐carbohydrate diet affects β‐cell mass (BCM) has been little investigated.Materials and MethodsBoth wild‐type (WT) mice and adenosine triphosphate‐sensitive potassium channel‐deficient (Kir6.2KO) mice were fed normal chow or high‐starch (ST) diets for 22 weeks. BCM and the numbers of islets were analyzed by immunohistochemistry, and gene expression levels in islets were investigated by quantitative real‐time reverse transcription polymerase chain reaction. MIN6‐K8 β‐cells were stimulated in solution containing various concentrations of glucose combined with nifedipine and glimepiride, and gene expression was analyzed.ResultsBoth WT and Kir6.2KO mice fed ST showed hyperinsulinemia and body weight gain. BCM, the number of islets and the expression levels of cyclinD2 messenger ribonucleic acid were increased in WT mice fed ST compared with those in WT mice fed normal chow. In contrast, no significant difference in BCM, the number of islets or the expression levels of cyclinD2 messenger ribonucleic acid were observed between Kir6.2KO mice fed normal chow and those fed ST. Incubation of MIN6‐K8 β‐cells in high‐glucose media or with glimepiride increased cyclinD2 expression, whereas nifedipine attenuated a high‐glucose‐induced increase in cyclinD2 expression.ConclusionsThese results show that a high‐starch diet increases BCM in an adenosine triphosphate‐sensitive potassium channel‐dependent manner, which is mediated through upregulation of cyclinD2 expression.

Dill-normalized liver lipid accumulation, oxidative stress, and low-density lipoprotein receptor levels in high cholesterol fed hamsters.

BACKGROUNDHigh lipid accumulation in hepatocyte and blood vessels can lead to non-alcoholic fatty liver disease (NAFLD) and heart diseases, respectively. These disorders are the main reasons of mortality in various countries. In this experiment, we evaluated the effect of leaf extracts of Anethum graveolens (AG), also known as Dill, and AG tablet on expression of low-density lipoprotein receptor (LDLR) and liver lipid in hypercholesterolemic hamsters.METHODSIn this experimental study, 36 male golden hamsters were divided into 6 groups: 1) standard diet + 0.5% cholic acid + 2% cholesterol [high cholesterol diet (HCD)], 2) HCD + 100 mg/kg hydroalcoholic extract of Dill, 3) HCD + 200 mg/kg hydroalcoholic extract of Dill, 4) HCD + 100 mg/kg Dill tablet, 5) HCD + 200 mg/kg Dill tablet, 6) chow. At the end of study (30th day), hamsters were anesthetized and blood sample and liver tissue were collected. Biochemical factors and antioxidant parameters were determined. LDLR messenger ribonucleic acid (mRNA) level was measured using real time polymerase chain reaction (RT-PCR). Histopathological change of liver was determined using light microscope.RESULTSCompared to HCD group, blood lipids (P < 0.0010) and liver enzymes (P < 0.0010) markedly reduced in AG-treated groups. The expression of LDLR did not change significantly in animals which received low dose of hydroalcoholic extract or AG tablet, but it increased in animals receiving high dose of extract or tablet (P < 0.0100). Liver antioxidant significantly increased by AG (P < 0.0010). Liver histopathological changes were normalized by AG.CONCLUSIONAG can significantly increase LDLR gene expression in HCD animals. This study showed that both AG extract and AG tablet had potential antioxidant and hypolipidemic effects in hamsters.

Antiproliferative potential of miR-33a in laryngeal cancer Hep-2 cells via targeting PIM1.

Laryngeal cancer is a frequent cause of cancer-associated mortality worldwide with an overall poor prognosis along with high mortality rates. Therefore, comprehensive investigation of underlying molecular mechanisms of laryngeal carcinogenesis remains an important problem. In this study, proliferative and apoptotic features of Hep-2 cells overexpressing microRNA-33a (miR-33a) were evaluated and in silico analysis along with literature search was used to find putative targets of miR-33a. The potential of PIM1 (pim-1 oncogene) as a direct target of miR-33a was tested using quantitative real-time polymerase chain reaction, Western blot, and luciferase assay. Induced miR-33a expression significantly inhibited proliferation through inducing apoptosis of Hep-2 cells. Further in vitro tests showed downregulation of PIM1 in messenger ribonucleic acid (mRNA) and protein level upon miR-33a overexpression and confirmed PIM1 as a direct target of miR-33a. Mir-33a was demonstrated to act as a tumor suppressor in larnygeal cancer via directly targeting the 3' untranslated region of PIM1.

In vitro anti-inflammatory and free radical scavenging properties of Oldenlandia auricularia

The in vitro anti-inflammatory and antioxidant properties of extract of Oldenlandia auricularia with 70% aqueous alcohol have been evaluated. The inhibition of nuclear translocation of nuclear factor kappa-light-chain-enhancer of activated B cells in lipopolysaccharide stimulated murine macrophage cells was evaluated. Messenger ribonucleic acid levels of tumor necrosis factor-α and cyclooxygenase-2 were analysed by quantitative real time polymerase chain reaction. O. auricularia (100 mg/L) showed better anti-inflammatory and antioxidant activities when compared to its lower doses, whereas the standard drugs curcumin and diclofenac sodium showed significant anti-inflammatory and antioxidant potential. Treatment at this concentration inhibited the signalling pathway of nuclear factor kappa-light-chain-enhancer of activated B cells and messenger ribonucleic acid expression of tumor necrosis factor- α, and cyclooxygenase-2. Liquid chromatography/quadrupole time-of-flight mass spectrometry revealed the presence of previously reported compounds auricularine, ursolic acid and β-sitosterol. The observed anti-inflammatory property of O. auricularia could be due to the inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells p65 sub unit nuclear translocation and inhibition of its signalling pathway.

Expression of c-Myc and Beclin-1 in skin of rats after burn.

This study aimed to investigate the expression features of Beclin-1 and c-Myc in the skin burn of rats. A total of 48 Sprague-Dawley (SD) rats were randomly divided into the normal group (n=12), the 3-day burn group (n=12), the 5-day burn group (n=12) and the 7-day burn group (n=12). Except for the normal group, the rat models of burn were established in the other three groups, burn wounds were given routine dressing change, and rats were sacrificed at 3, 5 and 7 days after modeling to collect materials. Then, immunohistochemistry was applied to detect the expression of c-Myc and Beclin-1. The expression levels of c-Myc protein and Beclin-1 protein were measured via western blotting. The expression levels of c-Myc messenger ribonucleic acid (mRNA) and Beclin-1 mRNA were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In comparison with the normal group, three burn groups had significantly increased the expression of c-Myc and Beclin-1, and the differences were statistically significant (P<0.05). Beclin-1 expression in the 5-day burn group was obviously higher than those in the 3 and 7-day burn groups, and the differences were of statistical significance (P<0.05). The expression of c-Myc in the 7-day burn group was overtly higher than those in the 3 and 5-day burn groups, and the differences showed statistical significance (P<0.05). The expression of Beclin-1 and c-Myc in post-burn skin tissues were gradually increased, with the Beclin-1 expression level reaching the peak on the 5th day after burn, and the expression level of c-Myc was the highest on the 7th day after burn.

Association Between Superoxide Dismutase Isoenzyme Gene Expression and Total Antioxidant Status in Patients with an End-Stage Renal Disease

Background:Chronic renal failure, particularly end-stage renal disease, is a serious health problem associated with a high mortality rate. Uremic syndrome leads to increased oxidative stress, inflammation, and dyslipidemia.Aims:To examine superoxide dismutase isoenzyme gene expression in peripheral blood mononuclear cells of patients on hemodialysis and to determine the associations between superoxide dismutase isoenzyme gene expression, oxidative stress, and non-enzymatic antioxidative protection.Study Design:Case control study.Methods:This study included 33 patients on hemodialysis (age, 55.33±15.31 years old) and 33 apparently healthy controls (age, 45.37±8.92 years old). Superoxide dismutase isoenzyme messenger ribonucleic acid levels were determined by real-time polymerase chain reaction. General biochemical parameters, high sensitivity C-reactive protein, total antioxidant status, thiobarbituric acid-reactive substances, and the superoxide anion radical were also determined.Results:Normalized Cu/Zn superoxide dismutase and Mn superoxide dismutase messenger ribonucleic acid levels were significantly higher in patients than controls (p<0.001 and p=0.011). A significant negative correlation was detected between normalized Cu/Zn superoxide dismutase messenger ribonucleic acid levels and total protein, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and total antioxidant status. Normalized Mn superoxide dismutase messenger ribonucleic acid levels were negatively correlated with total protein and total antioxidant status. A multiple regression analysis revealed independent associations between total antioxidant status and normalized Cu/Zn superoxide dismutase (p=0.038) and between total antioxidant status and normalized Mn superoxide dismutase messenger ribonucleic acid levels (p=0.038 and p=0.018, respectively).Conclusion:The superoxide dismutase isoenzyme gene is expressed at a higher rate in patients with end-stage renal failure, probably due to increased oxidative stress and attenuated antioxidative defense. The plasma total antioxidant status is an independent predictor of normalized superoxide dismutase isoenzyme messenger ribonucleic acid levels.

Biofabrication of a novel leukocyte-fibrin-platelet membrane as a cells and growth factors delivery platform for tissue engineering applications.

Autologous platelet-rich hemocomponents have emerged as potential biologic tools for regenerative purpose, but their therapeutic efficacy still remains controversial. This work represents the characterization study of an innovative autologous leukocyte-fibrin-platelet membrane (LFPm), which we prepared according to a novel protocol involving multiple cycles of apheresis. The high content in fibrinogen gave to our hemocomponent the appearance of a manipulable and suturable membrane with high elasticity and deformation capacity. Moreover, being highly enriched with platelets, leukocytes, and monocytes/macrophages, the LFPm sustained the local release of bioactive molecules (platelet derived growth factor, vascular endothelial growth factor, interleukin-10, and tumour necrosis factor alpha). In parallel, the evaluation of stemness potential highlighted also that the LFPm contained cells expressing pluripotency and multipotency markers both at the messenger ribonucleic acid (NANOG, SOX2, THY1, NT5E, and ENG) and surface-protein level (CD44high /CD73+ /CD34+ /CD117+ /CD31+ ). Finally, biodegradation analysis interestingly showed a good stability of the membrane for at least 3weeks in vitro and 1week in vivo. In both cases, biodegradation was associated with progressive exposure of fibrin scaffold, loss/migration of cellular elements, and release of growth factors. Overall, collected evidence could shed some light on the regenerative effect that LFPms may exert after the autologous implant on a defect site.

Fli-1 Governs Pericyte Dysfunction in a Murine Model of Sepsis.

Pericytes are vascular mural cells and are embedded in the basement membrane of the microvasculature. Recent studies suggest a role for pericytes in lipopolysaccharide (LPS)-induced microvascular dysfunction and mortality, but the mechanisms of pericyte loss in sepsis are largely unknown. By using a cecal ligation and puncture (CLP)-induced murine model of sepsis, we observed that CLP led to lung and renal pericyte loss and reduced lung pericyte density and pericyte/endothelial cell (EC) coverage. Up-regulated Friend leukemia virus integration 1 (Fli-1) messenger ribonucleic acid (RNA) and protein levels were found in lung pericytes from CLP mice in vivo and in LPS-stimulated lung pericytes in vitro. Knockout of Fli-1 in Foxd1-derived pericytes prevented CLP-induced pericyte loss, vascular leak, and improved survival. Disrupted Fli-1 expression by small interfering RNA inhibited LPS-induced inflammatory cytokines and chemokines in cultured lung pericytes. Furthermore, CLP-induced pericyte pyroptosis was mitigated in pericyte Fli-1 knockout mice. Our findings suggest that Fli-1 is a potential therapeutic target in sepsis.

Upregulation of High Mobility Group Box 1 May Contribute to the Pathogenesis of Biliary Atresia.

Biliary atresia (BA) is a progressive inflammatory obstructive cholangiopathy in infants. High mobility group box 1 (HMGB1) is known to play an important role as a late mediator of inflammation. However, it is not clear whether HMGB1 levels are of clinical significance in patients with BA. The aim of this study was to determine correlations between serum HMGB1 levels and the clinicopathologic features of BA. Serum samples were collected from 19 infants with BA, 7 infants with anicteric choledochal cysts (CC) and normal liver function, and 8 healthy controls. Serum HMGB1 levels were measured with an enzyme-linked immunosorbent assay. Routine liver function tests were performed on serum samples. Quantitative real-time polymerase chain reaction and western blot analyses were used to detect HMGB1 expression in BA liver biopsy tissues. Localization of HMGB1 expression in the hepatic lobule was determined by immunohistochemical analysis. HMGB1 levels in serum collected from BA infants were significantly elevated compared with CC and healthy control patients. Furthermore, elevated serum levels of HMGB1 in BA infants positively correlated with gamma-glutamyl transferase levels. HMGB1 messenger ribonucleic acid and protein expression levels were upregulated in BA liver biopsy tissues compared with CC patients. Immunohistochemical analysis also revealed increased positive immunostaining for HMGB1 in BA liver tissues as compared with CC tissues. HMGB1 may play a crucial role in the pathogenesis of BA. Additionally, the correlation of serum HMGB1 levels with gamma-glutamyl transferase levels may provide a novel marker for the diagnosis of BA.

Decreased placental and muscular expression of the fibroblast growth factor 19 in gestational diabetes mellitus.

Aims/IntroductionFibroblast growth factor (FGF)19 has been shown to improve glycemic homeostasis and lipid metabolism in animal models. In humans, decreased FGF19 level has been described in diabetes. The present study aimed to investigate the expression of FGF19 in gestational diabetes mellitus (GDM) patients.Materials and MethodsSamples for measurement were obtained from 20 women with GDM and 25 healthy controls. The messenger ribonucleic acid (mRNA) and protein expression levels of FGF19, FGF21 and co‐receptor β‐klotho (KLB) in the placenta, rectus muscle and subcutaneous fat tissues were quantified by real‐time quantitative polymerase chain reaction, western blot and immunohistochemistry, respectively.ResultsWomen with GDM had significantly lower mRNA and protein expressions of FGF19 than control women in the placenta (mRNA 0.33 ± 0.05 vs 0.72 ± 0.09; protein 0.34 ± 0.13 vs 0.85 ± 0.20) and rectus muscle (mRNA 0.83 ± 0.11 vs 1.28 ± 0.19; protein 0.78 ± 0.24 vs 1.23 ± 0.39). However, there were no significant differences between GDM women and controls with respect to the expression levels of FGF21 and β‐klotho in the placenta and rectus muscle. There were almost no detectable FGF19 and FGF21 expressions in subcutaneous fat tissue. Furthermore, β‐klotho expression levels were not different between the GDM and control group in subcutaneous fat.Conclusions FGF19 expressions are decreased in the placenta and rectus muscle of women with GDM. This might contribute to the pathophysiology or development of GDM.

Correlation between Nurr1 expression and drug resistance in the brain of rats with epilepsy.

To investigate the correlation between nuclear receptor related 1 (Nurr1) expression and drug resistance in the brain of rats with epilepsy. A total of 60 adult male Sprague-Dawley rats were selected, and the animal model of epilepsy was established by electrical stimulation. These rats were randomly divided into the control group and the drug-resistant group. The model of drug-resistant epilepsy was screened with phenytoin (PHT) and phenobarbital (PB); the hippocampus and temporal lobe cortex tissues were isolated from rats; the messenger ribonucleic acid (mRNA) and protein levels of Nurr1 in the hippocampus and cortex tissues of the two groups of rats were detected by reverse transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry, respectively. The budding tissue marker growth-associated protein-43 (GAP43) in hippocampus tissues were labeled by immunofluorescence staining; the mRNA and protein levels of indicators related to drug-resistant epilepsy, including leukemia-associated phenotype (LAP), multi-drug resistance protein (MRP), P-glycoprotein (P-gp) and monocyte chemoattractant protein-1 (MCP-1), were further detected. Pearson correlation analysis was used to analyze the correlations of the protein level of Nurr1 with drug-resistant indicators. The mRNA and protein levels of Nurr1 in hippocampus tissues of the drug-resistant group were significantly increased compared with those of the control group (p < 0.05). In temporal lobe cortex tissues, there was no significant difference in the mRNA level of Nurr1 between the control group and the drug-resistant group (p > 0.05). The immunohistochemistry results showed that the fluorescence intensity of GAP3 in hippocampus tissues of the drug-resistant group was significantly higher than that of the control group. The mRNA and protein levels of epilepsy-related indicators, LAP, MRP, P-gp and MCP-1 in hippocampus tissues of drug-resistant group were significantly up-regulated compared with those of the control group (p < 0.05). Pearson correlation analysis indicated that the protein level of Nurr1 in hippocampus tissues was positively correlated with those of LAP, MRP, P-gp and MCP-1 in the corresponding regions (p < 0.05). Nurr1 enhances the drug resistance of epilepsy in rats by up-regulating the expression of proteins related to drug resistance.

Ultrasound Irradiation Combined with Hepatocyte Growth Factor Accelerate the Hepatic Differentiation of Human Bone Marrow Mesenchymal Stem Cells

This study investigated the impact of ultrasound (US) irradiation on the hepatic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) induced by hepatocyte growth factor (HGF) and the possible mechanisms. We treated hBMSCs, using HGF with and without US irradiation. Cell viability and stem cell surface markers were analyzed. Hepatocyte-like cell markers and functional markers including α-fetoprotein (αFP/AFP), cytokeratin 18 (CK18), albumin (ALB) and glycogen content were analyzed at the time point of day 1, 3 and 5 after treatment. The involvement of Wnt/β-catenin signaling pathway was evaluated as well. The results showed that the US treatment at 1.0 W/cm2 or 1.5 W/cm2 for 30 s or 60 s conditions yielded favorable cell viability and engendered stem cell differentiation. At day 5, the expressions of AFP, CK18, ALB and the glycogen content were significantly elevated in the US-treated group at both messenger ribonucleic acid and protein levels (all p < 0.05), in comparison with HGF and control groups. Among all the US treated groups, the expression levels of specific hepatic markers in the (1.5 W/cm2 for 60 s) group were the highest. Furthermore, Wnt1, β-Catenin, c-Myc and Cyclin D1 were significantly increased after US irradiation (all p < 0.05), and the enhancements of c-Myc and Cyclin D1 could be obviously impaired by the inhibitor ICG-001 (p < 0.05, p < 0.05), in accordance with decreased ALB and CK18 expression and glycogen content (all p < 0.05). In conclusion, US irradiation was able to promote the hBMSCs' differentiation mediated by HGF in vitro safely, easily and controllably. The activation of Wnt/β-catenin signaling pathway was involved in this process. US irradiation could serve as a potentially beneficial tool for the research and application of stem cell differentiation.

Effects of atorvastatin on autophagy in skeletal muscles of diabetic rats.

Aims/IntroductionAtorvastatin is usually used to decrease the amount of fatty substances in individuals with type 2 diabetes mellitus. However, it can cause side‐effects, such as breakdown of skeletal muscle tissue. The present study focused on the effects of atorvastatin on autophagy of the skeletal muscles in diabetic rats.Materials and MethodsDiabetes in rats in the diabetic (D) and atorvastatin (T) groups was induced using streptozotocin (65 mg/kg, intraperitoneal injection). Next, rats in the T group were treated with atorvastatin (10 mg/kg/day, intragastric administration), whereas rats in the control and D groups were given water. Additionally, the rats in T and D groups were fed a high‐fat and high‐sugar diet for 10 weeks. Subsequently, the histopathological changes, and expression levels of microtubule‐associated protein 1 light chain 3 (LC3)‐I/‐II and p62 in the skeletal muscle specimens in the three groups were analyzed.ResultsRats in the T group had reduced lipid droplets, cholesterol and low‐density lipoprotein (P < 0.05) levels than those in the D group. Disordered atrophic myocytes, incrassated vascular walls and decreased cross‐sectional area of type I fibers were found using hematoxylin–eosin and adenosine triphosphatase staining in the D and T groups. The messenger ribonucleic acid and protein levels of LC3‐II and the LC3‐II/LC3‐I ratio were increased in the T group compared with those in the other groups (P < 0.05), whereas the protein level of p62 showed the opposite trend.ConclusionsAtorvastatin enhanced the autophagy level of skeletal muscles to decrease lipid deposition, which possibly exacerbated myopathy.

Analysis of the levels of Th9 cells and cytokines in the peripheral blood of mice with bronchial asthma.

The purpose of the study was to detect the level of T-helper type 9 (Th9) cells and the cytokine interleukin-9 (IL-9) in peripheral blood of mice with bronchial asthma, and to explore the relationship between the expression of Th9 cells and the pathogenesis of asthma. Thirty female-specific pathogen-free (SPF) Bagg' albino (BALB)/c mice were selected and randomly divided into the control group (n=15) and the bronchial observation group (n=15). Mice in the bronchial observation group were treated with ovalbumin (OVA) for sensitization and induction of a mouse model of asthma. The airway reactivity of mice was measured by a mouse pulmonary function apparatus using the non-invasive pulmonary impedance method. The proportions of Th9 cells in peripheral blood of mice in the two groups were detected using flow cytometry. Digital polymerase chain reaction (dPCR), enzyme-linked immunosorbent assay (ELISA) and western blot analysis were applied to detect the levels of IL-9 messenger ribonucleic acid (mRNA) and proteins in peripheral blood and lung tissues of mice in the two groups, respectively. Compared with that in the control group, the expression level of Th9 cells in the peripheral blood of mice in the observation group was significantly elevated (P<0.05), the expression level of IL-9 proteins in the peripheral blood was significantly increased (P<0.05), and the levels of IL-9 mRNA and proteins in lung tissues were significantly increased (P<0.05). The results show that the levels of Th9 cells and their cytokine IL-9 in the peripheral blood of mice with bronchial asthma are significantly increased, suggesting that Th9 cells play important roles in the pathogenesis of asthma.

Analysis of the association of the expression of KiSS-1 in colorectal cancer tissues with the pathology and prognosis.

Colorectal cancer is a common malignant tumor of the digestive tract with high morbidity and mortality rates. The aim of the present study was to examine the expression level of KiSS-1 in tumor tissues of patients with colorectal cancer, and to explore the relationship with the clinicopathology and prognosis of patients with colorectal cancer. Frozen tumor tissue and corresponding cancer-adjacent normal tissue specimens were selected from 56 patients with colorectal cancer who were treated in the Department of Surgery of our hospital from May 2009 to December 2011. The expression levels of KiSS-1 messenger ribonucleic acid (mRNA) in tumor tissues and cancer-adjacent normal tissues were detected by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). The expression levels of KiSS-1 proteins in colorectal cancer tissues and cancer-adjacent normal tissues were further detected by immunohistochemistry. In addition, the association of the expression level of KiSS-1 proteins in tissues of colorectal cancer patients with pathological parameters and the prognosis of patients with colorectal cancer was analyzed combined with clinical data. The RT-qPCR results showed that the expression of KiSS-1 mRNA in colorectal cancer tissues was significantly lower than that in cancer-adjacent normal tissues (P<0.05). Immunohistochemistry results indicated that the positive expression rate of KiSS-1 proteins in colorectal cancer tissues (26.79%) was significantly lower than that in cancer-adjacent normal tissues (80.36%). The low expression of KiSS-1 in colorectal cancer tissues was associated with the degree of differentiation, invasion and metastasis, as well as clinical staging. The 5-year overall survival rate of patients with colorectal cancer was 55.36% (31/56). The univariate survival analysis showed that patients with lowly expressed KiSS-1 had worse prognosis. The low expression of KiSS-1 is closely associated with the occurrence and development of colorectal cancer, especially to the degree of differentiation, invasion and metastasis, as well as clinical staging. Thus, the expression of KiSS-1 in colorectal cancer tissues can be used as a reference for the prognosis of colorectal cancer, and KiSS-1 is a potential new target for the treatment of colorectal cancer.

Depletion of CD11c+ Cells Does Not Influence Outcomes in Mice Subjected to Transient Middle Cerebral Artery Occlusion

Objective: While it has been shown that different T-cell subsets have a detrimental role in the acute phase of ischemic stroke, data on the impact of dendritic cells (DC) are missing. Classic DC can be characterized by the cluster of differentiation (CD)11c surface antigen. Methods: In this study, we depleted CD11c⁺ cells by using a CD11c-diphtheria toxin (DTX) receptor mouse strain that allows selective depletion of CD11c⁺ cells by DTX injection. For stroke induction, we used the model of transient middle cerebral artery occlusion (tMCAO) and analyzed stroke volume and functional outcome on days 1 and 3 as well as expression of prototypical pro- and anti-inflammatory cytokines on day 1 after tMCAO. Three different protocols for CD11c⁺ cell depletion, tMCAO duration, and readout time point were applied. Results: Injection of DTX (5 or 100 ng/g) reliably depleted CD11c⁺ cells without influencing the fractions of other immune cell subsets. CD11c⁺ cell depletion had no impact on stroke volume, but mice with a longer DTX pretreatment performed worse than those with vehicle treatment. CD11c⁺ cell depletion led to a decrease in cortical interleukin (IL)-1β and IL-6 messenger ribonucleic acid levels. Conclusions: We show, for the first time, that CD11c⁺ cell depletion does not influence stroke volume in a mouse model of focal cerebral ischemia. Nevertheless, given the unspecificity of the CD11c surface antigen for DC, mouse models that allow a more selective depletion of DC are needed to investigate the role of DC in stroke pathophysiology.

Prenatal Depression and Infant Temperament: The Moderating Role of Placental Gene Expression.

Prior research has demonstrated the link between maternal depression during pregnancy (i.e., prenatal depression) and increased neurodevelopmental dysregulation in offspring. However, little is known about the roles of key hypothalamic-pituitary axis regulatory genes in the placenta modulating this association. This study will examine whether placental gene expression levels of 11β-hydroxysteroid dehydrogenase type 2 (HSD11B2), glucocorticoid receptor (NR3C1), and mineralocorticoid receptor (NR3C2) can help elucidate the underlying mechanisms linking prenatal depression to infant temperament, particularly in infants with high negativity and low emotion regulation. Stored placenta tissues (N = 153) were used to quantify messenger ribonucleic acid levels of HSD11B2, NR3C1, and NR3C2. Assessments of prenatal depression and infant temperament at 6 months of age were ascertained via maternal report. Results found that prenatal depression was associated with increased Negative Affectivity (p < .05) after controlling for postnatal depression and psychosocial characteristics. Furthermore, the association between prenatal depression and Negative Affectivity was moderated by gene expression levels of HSD11B2, NR3C1, and NR3C2 such that greater gene expression significantly lessened the association between prenatal depression and Negative Affectivity. Our findings suggest that individual differences in placental gene expression may be used as an early marker of susceptibility or resilience to prenatal adversity.

The Effect of Functional Mandibular Shift on the Muscle Spindle Systems in Head-Neck Muscles and the Related Neurotransmitter Histamine.

The aim of this study is to explore the effects of abnormal occlusion and functional recovery caused by functional mandible deviation on the head and neck muscles and muscle spindle sensory-motor system by electrophysiological response and endogenous monoamine neurotransmitters’ distribution in the nucleus of the spinal tract. Seven-week-old male Wistar rats were randomly divided into 7 groups: normal control group, 2W experimental control group, 2W functional mandible deviation group, 2W functional mandible deviation recovery group, 4W experimental control group, 4W functional mandible deviation group, 4W functional mandible deviation recovery group. Chewing muscles, digastric muscle, splenius, and trapezius muscle spindles electrophysiological response activities at the opening and closing state were recorded. And then the chewing muscles, digastric, splenius, trapezius, and neck trigeminal nucleus were taken for histidine decarboxylase (HDC) detection by high performance liquid chromatography (HPLC), immunofluorescence, and reverse-transcription polymerase chain reaction (RT-PCR). Histamine receptor proteins in the neck nucleus of the spinal tract were also examined by immunofluorescence and RT-PCR. Electromyography activity of chewing muscles, digastric, and splenius muscle was significantly asymmetric; the abnormal muscle electromyography activity was mainly detected at the ipsilateral side. After functional mandibular deviation, muscle sensitivity on the ipsilateral sides of the chewing muscle and splenius decreased, muscle excitement weakened, modulation depth decreased, and the muscle spindle afferent impulses of excitation transmission speed slowed down. Changes for digastric muscle electrical activity were contrary. The functions recovered at different extents after removing the deflector. However, trapezius in all the experimental groups and recovery groups exhibited bilateral symmetry electrophysiological responses, and no significant difference compared with the control group. After functional mandibular deviation, HDC protein and messenger ribonucleic acid (mRNA) levels on the ipsilateral sides of the chewing muscle and splenius increased significantly. HDC level changes for digastric muscle were contrary. After the removal of the mandibular position deflector, HDC protein and mRNA levels decreased on the ipsilateral sides of the chewing muscle and splenius while they increased in the digastric muscle. The difference of histamine decarboxylase content in the bilateral trapezius in each experimental group was small. After functional mandibular deviation, the temporomandibular joint mechanical receptors not only caused the fusimotor fiber hypoallergenic fatigue slow response on the ipsilateral sides of splenius, but also increased the injury neurotransmitter histamine release. The authors’ results further support the opinion that the temporomandibular joint receptors may be involved in the mechanical theory of the head and neck muscles nervous system regulation.

Chronic exposure of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces an obesogenic effect in C57BL/6J mice fed a high fat diet

Contaminant involvement in the pathophysiology of obesity is widely recognized. It has been shown that low dose and chronic exposure to endocrine disruptor compounds (EDCs) potentiated diet- induced obesity. High and acute exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a persistent organic pollutant (POP) and an EDC with anti-estrogenic property, causes wasting syndrome . However at lower doses, the TCDD metabolic effects remain poorly understood. We investigated the obesogenic effect during chronic exposure of TCDD at 1μg/kg body weight (bw)/week in adult C57BL/6J mice fed with a high fat diet (HFD) and exposed from 10 to 42 weeks old to TCDD or equal volume of vehicle by intragastric gavage. Under these conditions, TCDD was obesogenic in adult mice (7% in males and 8% in females), which was linked to fat mass. A sex effect was observed in the fat mass distribution in adipose tissue and in the hepatic triglyceride content evolution. In visceral fat pad weight, we observed a decrease (11%) in males and an increase (14%) in females. The hepatic triglyceride content increase (41%) in females only. TCDD failed to induce any change in plasma parameters regarding glucose and lipid homeostasis. Messenger ribonucleic acid (mRNA) levels involved in adipose tissue and hepatic metabolism, inflammation, xenobiotic metabolism and endocrine disruption were differently regulated between males and females. In conclusion, these results provide new evidence that dioxin, a POP and EDC can be obesogenic for adult mice with multi-organ effects.

Downregulated Elastin Microfibril Interfacer 1 Expression in the Pulmonary Vasculature of Experimental Congenital Diaphragmatic Hernia.

Pulmonary hypertension (PH) is a severe complication of congenital diaphragmatic hernia (CDH). Transforming growth factor-β (TGFβ) signaling is suggested to be involved in PH development by regulating embryonic angiogenesis, cell proliferation, and cell differentiation. Altered TGFβ signaling has been demonstrated in experimental CDH lungs. Elastin microfibril interfacer 1 (Emilin-1) is an extracellular matrix glycoprotein expressed in endothelial and vascular smooth muscle cells and known to regulate TGFβ processing and arterial diameter. We designed this study to investigate the pulmonary vascular expression of Emilin-1 in nitrofen-induced CDH rats. Following ethical approval (REC913b, REC1103), time-pregnant Sprague Dawley rats received nitrofen or vehicle on gestational day 9 (D9). Fetuses were sacrificed on D21 and divided into CDH group and control group. Quantitative real-time polymerase chain reaction (n = 11 each group), Western blot analysis, and confocal microscopy were used to determine the gene and protein expression of Emilin-1. Relative Emilin-1 messenger RNA (ribonucleic acid) levels were significantly downregulated in CDH lung tissue compared with controls (CDH: 0.043 ± 0.003; control: 0.067 ± 0.004; p < 0.001). Western blotting confirmed the decreased pulmonary Emilin-1 protein expression in CDH lungs. Confocal microscopy demonstrated a markedly diminished expression of Emilin-1 in the CDH pulmonary vasculature compared with controls. To our knowledge, this study demonstrates for the first time a decreased Emilin-1 gene and protein expression in the pulmonary vasculature of nitrofen-induced CDH. Emilin-1 deficiency through its interaction with TGFß may result in abnormal vascular remodeling resulting in PH in this model.