Binding Protein Levels Research Articles

High density lipoprotein inhibited group II innate lymphoid cells proliferation and function in allergic rhinitis

BackgroundMore and more studies had suggested that dyslipidemia was closely related to allergic diseases. High density lipoprotein (HDL) often plays anti-inflammatory and anti-oxidative roles by suppressing immune cell chemotaxis and activation. We aimed to explore the role of HDL in the regulation of group II innate lymphoid cells (ILC2) in allergic rhinitis (AR).MethodsThe blood lipid levels and their correlation with symptom scores of 20 AR subjects and 20 controls were analyzed. Purified ILC2 were stimulated by HDL and cytokines production were examined by enzyme-linked immunosorbent assay (ELISA) and flow cytometry. The mRNA levels of GATA binding protein 3(GATA3) and retinoid-related orphan receptor α (RORα) expressed by ILC2 were detected using reverse transcription polymerase chain reaction (RT-PCR).ResultsHDL level was significantly lower in AR than controls and correlated with the symptom scores. The serum HDL levels were negatively related to the increased number of ILC2, IL-5+ ILC2, and IL-13+ ILC2 in AR patients. HDL decreased the number of ILC2 and type II cytokines levels significantly by inhibiting expression of GATA3 and RORα.ConclusionsOur data provide preliminary evidence that HDL may play a negative role in ILC2 inflammation in AR, suggesting that HDL may serve as promising treatment target in AR.

MO735: Increased Serum Adipocyte Fatty Acid Binding Protein Level Associated with Endothelial Dysfunction in Patients Undergoing Hemodialysis

Abstract BACKGROUND AND AIMS Adipocyte fatty acid binding protein (A-FABP) is independently associated with atherosclerosis. Endothelial dysfunction is one of the causes of cardiovascular morbidity and mortality in patients with hemodialysis (HD). This study evaluated the correlation between serum A-FABP level and endothelial function measured by vascular reactivity index (VRI) in patients with HD. METHOD Fasting blood samples were obtained from 90 HD patients. A commercial enzyme immunoassay kit was used to measure A-FABP levels. The endothelial function and VRI were measured using a digital thermal monitoring (DTM) test (VENDYS). In this study, VRI < 1.0 was used as the poor vascular reactivity, 1.0 ≤ VRI < 2.0 was used as the intermediate vascular reactivity, and VRI ≥ 2.0 was used as the good vascular reactivity. RESULTS A total of 14 HD patients (15.6%) were categorized as poor vascular reactivity (VRI < 1.0), 38 HD patients (42.2%) were categorized as intermediate vascular reactivity (1.0 ≤ VRI < 2.0) and 38 HD patients had good vascular reactivity. Decreased pre-HD body weight (P = 0.016), post-HD body weight (P = 0.013), body mass index (P = 0.034), serum creatinine level (P = 0.010) and higher serum A-FABP level (P = 0.001) were associated with poor vascular reactivity. After multivariable forward stepwise linear regression analysis noted that serum creatinine level (β = 0.234, adjusted R2 change = 0.043; P = 0.019) was positively associated with, and serum A-FABP level (β = −0.347, adjusted R2 change = 0.152; P = 0.001) was negatively associated with VRI values in patients undergoing HD. CONCLUSION After adjusting for significant confounders, the serum A-FABP level was negatively correlated, and the serum creatinine level was positively correlated with VRI values in patients undergoing HD.

MO172: Correlation of Serum Adipocyte Fatty Acid Binding Protein Level and Peripheral Arterial Disease in Chronic Kidney Patients

Abstract BACKGROUND AND AIMS Adipocyte fatty acid-binding protein (A-FABP) is expressed in adipocytes, dendritic cells and macrophages and it was thought to be involved in insulin resistance, chronic inflammation and atherosclerotic processes. Peripheral arterial disease (PAD) is associated with all-cause mortality, cardiovascular events and a key pathological process is atherosclerosis. The present study aimed to determine the relationship between serum A-FABP level and PAD in patients with non-dialysis chronic kidney disease (CKD). METHOD Fasting blood samples and baseline characteristics were obtained from 260 non-dialysis CKD patients (CKD stage 1–5). Ankle-brachial index (ABI) values were measured using an automated oscillometric device. Patients with ABIs of <0.9 were categorized into the low ABI group. Serum A-FABP levels were measured using a commercial enzyme immunoassay assay. RESULTS In the study cohort, 32 of the 260 non-dialysis CKD patients (12.3%) had low ABIs. The rates of diabetes mellitus (P = .037), smoking (P = .030), older age (P = .002) as well as the serum levels of A-FABP (P = .003) were higher in the low ABI group compared with the normal ABI group. The multivariable logistic regression analysis revealed that serum levels of A-FABP [odds ratio (OR): 1.223, 95% confidence interval (CI): 1.068–1.401, P = .004], older age (OR: 1.064, 95% CI: 1.026–1.104, P = .001) and smoking (OR: 4.438, 95% CI: 1.493–13.198, P = .007) were the independently associated with PAD in patients with CKD. By receiver operating characteristic (ROC) curve analysis, the area under the receiver operating characteristic curve of A-FABP level was 0.662 (95% CI: 0.601–0.719, P = .0004) for predicting PAD in non-dialysis CKD patients. CONCLUSION In this study, serum A-FABP levels were associated with PAD in patients with non-dialysis CKD.

Physiological Changes in Response to Skeletal Muscle Specific High Expression of mu‐Crystallin Following High and Low‐Fat Diets

Approximately 20 percent of the population expresses high levels of the thyroid hormone binding protein mu‐crystallin in skeletal muscle while the rest of the population expresses little to no mu‐crystallin. Since the thyroid hormones, T3 and T4, are potent regulators of metabolism and thermogenic regulation, our studies focus on the consequences of skeletal‐specific high expression (SM‐HE) of mu‐crystallin. We hypothesize that SM‐HE of mu‐crystallin influences cellular and metabolic changes in a diet‐ and sex‐ specific manner by influencing changes in gene expression.To evaluate this, we generated transgenic mice (Crym‐tg) that over‐express mu‐crystallin in skeletal muscle, equivalent to human high expressors, which resulted in high levels of accumulation of T3 in muscle tissue. Respiratory exchange ratio (RER) in these mice indicated a small but significant increase in utilization of fat as an energy source.In high‐ and low‐fat diet studies, we found differences between Crym‐tg and controls as well as sex specific differences. Following a 16‐week exposure, Crym‐tg males on high‐fat diet and Crym‐tg females on low‐fat diet weighed more than controls whereas Crym‐tg males on low‐fat diet and Crym‐tg females on high‐fat diet weighed less than controls. These differences were consistent with the results of glucose tolerance tests, in which only the Crym‐tg mice that weighed more than controls showed a significant increase in sensitivity to glucose. We are currently evaluating these results to understand their basis in transcriptomic and proteomic data, which have already revealed increased expression of genes involved in fatty acid oxidation and decreased expression of genes involved in glycolytic metabolism (Kinney et al. (2021) Curr Res Physiol. 4:47‐59.) as well as some members of the insulin signaling pathway.

Effects of spliceosomal mutations on brain patterning and morphogenesis

The major spliceosome consists of U1, U2, U5, and U4/U6 small nuclear ribonucleoprotein (snRNPs), and each snRNP has distinct and sequential roles during the mRNA splicing process. Mutations in the core components of the spliceosome are associated with brain and neurological defects. The goal of this study is to examine the role of three core spliceosomal components, SNRPB, SF3B4, and EFTUD2, in brain patterning and morphogenesis, and whether or not they are required in the neural tube or neural crest cells for brain development. Previously, we identified brain defects associated with the mutation of these spliceosomal components in the neural tube and neural crest cells. Structures derived from the midbrain and hindbrain region, such as the diencephalon, pons, and cerebellum, were found to be absent in E12.5 and E14.5 mutant embryos. In this study, we aim to use mutant mouse models to identify shared transcripts and pathways disrupted by mutations in these spliceosomal genes. Through RNA‐seq analysis, we identified a number of mis‐expressed RNA‐binding proteins that may contribute to the malformationis found in these mutants. We plan to examine level and expression of these RNA binding proteins in wild type and mutant embryos to evaluate how they contribute to the phenotypes found. Furthermore, to assess the potential clinical application of our findings, we will use human embryonic stem cells (hESC) to generate human neural crest cells with mutatioin in these splicing factors; shared transcripts and pathways identified in the mouse model will be compared to those found in induced human neural crest cells. Distinct spliceosomophathies are difficult to diagnose due to phenotypic overlaps. The significance of this work lies in its potential to identify a disrupted pathway shared by mutations in various splicing factors, a therapeutic alternative across multiple spliceosomopathies.

Abstract 106: Ribosomal Protein Rpl17 Controls Endothelial Cell Translation: A Key Mediator Of Intima Growth And Vascular Disease

Background: Carotid intima-media thickening (IMT) is a significant risk factor for cardiovascular disease (CVD). Using genetic analysis of the partial carotid ligation (PCL) model of disturbed flow induced IMT, we identified a strong association between decreased expression of the large ribosomal subunit protein 17 (RpL17) and increased IMT. RpL17 is located at the exit tunnel of the ribosome, making it critical for processing of newly translated proteins. To define the mechanism by which RpL17 inhibits IMT, we generated EC-restricted (Cdh5-Cre) conditional RpL17 heterozygous mice (EC-RpL17). Results: 1) PCL of EC-RpL17 increased IMT vs wild type (Control) littermates. 2) EM analysis revealed decreases in total ribosome number and polysomes in EC-RpL17, consistent with decreased global translation. 3) RNA-seq showed major changes in two pathways: Eukaryotic Initiation Factor 2 (eIF2α) that promotes adaptive translational responses to stress, and oxidative phosphorylation (OXPHOS), the electron transport metabolic pathway that produces ATP. 4) Profiling of mRNAs bound to polysomes in RpL17-EC revealed increased translational efficiency (TE) of selective redox regulators, pro-inflammatory mediators, matrix metalloproteases, and endothelial to mesenchymal transition (endMT) factors; consistent with processes that promote IMT. 5) Increased levels of RNA binding proteins (RBPs) in EC-RpL17, such as QKI, HuR, and hn-RNP, correlated with enhanced stability of specific mRNAs and their TE. 6) Increased TE of glycolytic enzymes and the lactate transporter was consistent with increased EC glycolysis and OXPHOS dysfunction. 7) Increased glycolysis correlated with greater cell proliferation, suggesting RpL17 is a growth suppressor. Conclusion: These data define a new mechanism for control of EC function via selective changes in mRNA stability and ribosome profile and number that decreased OXPHOS and global protein translation, leading to EC-proliferation and endMT, which enhanced IMT in the PCL model of disturbed flow.

Serum Mac-2 binding protein level predicts the development of liver-related events and colorectal cancer in patients with NAFLD.

We previously demonstrated that Mac‐2 binding protein (M2BP) is a useful biomarker for nonalcoholic fatty liver disease (NAFLD), particularly NAFLD fibrosis prediction. In the present study, we investigated the prognostic value of M2BP in patients with NAFLD. A total of 506 patients with biopsy‐confirmed NAFLD from 2002 to 2013 were enrolled in this study in Japan. Three hundred fifty‐three of these patients with NAFLD were available for follow‐up for more than 100 days and showed no liver‐related events at the time of entry. Liver‐related events were defined as hepatocellular carcinoma (HCC), decompensation, and gastroesophageal varices with variceal treatment. The mean follow‐up duration of all the subjects was 2716 ± 1621 days (102–7483 days). Eighteen patients developed new liver‐related events (HCC, 8; decompensation, 11; varices, 8). Nine patients developed cardiovascular disease (CVD), and 24 patients developed new cancers in other organs. The median serum M2BP level was 1.603 μg/mL, and we divided our cohort into two groups according to the serum M2BP level: M2BP low group (M2BP Low) and M2BP high group (M2BP Hi). The incidence of HCC was significantly higher in M2BP Hi (n = 8) than in M2BP Low (n = 0). The incidence of liver‐related events was significantly higher in M2BP Hi (n = 16) than in M2BP Low (n = 2). The incidences of death, CVD events, and cancer in other organs were not different between the groups. Interestingly, the incidence of colorectal cancer was significantly higher in M2BP Hi (n = 5) than in M2BP Low (n = 0). Conclusion: M2BP is a useful biomarker to predict liver‐related events, particularly HCC. Additionally, M2BP is a potential predictive biomarker of colorectal cancer development.

Declining Levels and Bioavailability of IGF-I in Cardiovascular Aging Associate With QT Prolongation-Results From the 1946 British Birth Cohort.

BackgroundAs people age, circulating levels of insulin-like growth factors (IGFs) and IGF binding protein 3 (IGFBP-3) decline. In rat cardiomyocytes, IGF-I has been shown to regulate sarcolemmal potassium channel activity and late sodium current thus impacting cardiac repolarization and the heart rate-corrected QT (QTc). However, the relationship between IGFs and IGFBP-3 with the QTc interval in humans, is unknown.ObjectivesTo examine the association of IGFs and IGFBP-3 with QTc interval in an older age population-based cohort.MethodsParticipants were from the 1946 Medical Research Council (MRC) National Survey of Health and Development (NSHD) British birth cohort. Biomarkers from blood samples at age 53 and 60–64 years (y, exposures) included IGF-I/II, IGFBP-3, IGF-I/IGFBP-3 ratio and the change (Δ) in marker levels between the 60–64 and 53y sampled timepoints. QTc (outcome) was recorded from electrocardiograms at the 60–64y timepoint. Generalized linear multivariable models with adjustments for relevant demographic and clinical factors, were used for complete-cases and repeated after multiple imputation.ResultsOne thousand four hundred forty-eight participants were included (48.3% men; QTc mean 414 ms interquartile range 26 ms). Univariate analysis revealed an association between low IGF-I and IGF-I/IGFBP-3 ratio at 60–64y with QTc prolongation [respectively: β −0.30 ms/nmol/L, (95% confidence intervals −0.44, −0.17), p < 0.001; β−28.9 ms/unit (-41.93, −15.50), p < 0.001], but not with IGF-II or IGFBP-3. No association with QTc was found for IGF biomarkers sampled at 53y, however both ΔIGF-I and ΔIGF-I/IGFBP-3 ratio were negatively associated with QTc [β −0.04 ms/nmol/L (−0.08, −0.008), p = 0.019; β −2.44 ms/unit (-4.17, −0.67), p = 0.007] while ΔIGF-II and ΔIGFBP-3 showed no association. In fully adjusted complete case and imputed models (reporting latter) low IGF-I and IGF-I/IGFBP-3 ratio at 60–64y [β −0.21 ms/nmol/L (−0.39, −0.04), p = 0.017; β −20.14 ms/unit (−36.28, −3.99), p = 0.015], steeper decline in ΔIGF-I [β −0.05 ms/nmol/L/10 years (−0.10, −0.002), p = 0.042] and shallower rise in ΔIGF-I/IGFBP-3 ratio over a decade [β −2.16 ms/unit/10 years (−4.23, −0.09), p = 0.041], were all independently associated with QTc prolongation. Independent associations with QTc were also confirmed for other previously known covariates: female sex [β 9.65 ms (6.65, 12.65), p < 0.001], increased left ventricular mass [β 0.04 ms/g (0.02, 0.06), p < 0.001] and blood potassium levels [β −5.70 ms/mmol/L (−10.23, −1.18) p = 0.014].ConclusionOver a decade, in an older age population-based cohort, declining levels and bioavailability of IGF-I associate with prolongation of the QTc interval. As QTc prolongation associates with increased risk for sudden death even in apparently healthy people, further research into the antiarrhythmic effects of IGF-I on cardiomyocytes is warranted.

Relationship between cerebrospinal fluid neurodegeneration biomarkers and temporal brain atrophy in cognitively healthy older adults

It is unclear whether cerebrospinal fluid (CSF) biomarkers of neurodegeneration predict brain atrophy in cognitively healthy older adults, whether these associations can be explained by phosphorylated tau181 (p-tau) and the 42 amino acid form of amyloid-β (Aβ42) biomarkers, and which neural substrates may drive these associations. We addressed these questions in 2 samples of cognitively healthy older adults who underwent longitudinal structural MRI up to 7 years and had baseline CSF levels of heart-type fatty-acid binding protein (FABP3)=, total-tau, neurogranin, and neurofilament light (NFL) (n = 189, scans = 721). The results showed that NFL, total-tau, and FABP3 predicted entorhinal thinning and hippocampal atrophy. Brain atrophy was not moderated by Aβ42 and the associations between NFL and FABP3 with brain atrophy were independent of p-tau. The spatial pattern of cortical atrophy associated with the biomarkers overlapped with neurogenetic profiles associated with expression in the axonal (total-tau, NFL) and dendritic (neurogranin) components. CSF biomarkers of neurodegeneration are useful for predicting specific features of brain atrophy in older adults, independently of amyloid and tau pathology biomarkers.

Mongolian Medicine Areca Thirteen Pill (GY-13) Improved Depressive Syndrome via upregulating cAMP/PKA/CREB/BDNF signaling pathway

Ethnopharmacological relevanceAreca Thirteen Pill, also called Gao You-13 (GY-13), is a traditional Mongolian herbal formula and has been extensively used to treat depression in Mongolian areas, which belongs to Heyi disease in Mongolian medicine. Major depressive disorder is a serious psychiatric disease, only one-third of individuals with depression are responsive to current antidepressants in clinic. Growing attention has been attracted by traditional herbal medicines in fighting depression because they are considered safer alternatives to pharmacotherapy. Aim of the studyTo reveal the mechanism of GY-13 in the treatment of depression. Materials and methodsThe rat depression model was established by chronic unpredictable mild stress (CUMS), and primary hippocampal neurons were used to construct a glutamate-induced excitotoxicity model. The antidepressant effect of GY-13 was then assessed by performing sucrose preference tests, open field tests, and body weight measurements on rats. The expression of cAMP and PKA, mRNA levels of brain-derived neurotrophic factor (BDNF) and cAMP response element binding protein (CREB), and hippocampal neuronal apoptosis were measured. ResultsThe results indicate that GY-13 significantly improves depression-like behavior, rescues decreased cAMP， PKA, recovers the mRNA levels of CREB and BDNF, and increases the proliferative activity of hippocampus. In addition, blockade of PKA reverses the effects of GY-13 treatment on CREB mRNA, BDNF mRNA levels. In vitro, GY-13 treatment increased hippocampal proliferative activity and attenuated Glu-induced apoptosis of hippocampal neurons as well as reduced CREB mRNA and BDNF mRNA expression levels. ConclusionsOur research demonstrated that GY-13 treatment exerted a potent antidepressant action via activation of cAMP/CREB/BDNF signaling pathway, promoting proliferation, and suppressing apoptosis. This research provides molecular biological ground for developing GY-13 into a potent alternative for the intervention of depression.

Identification of microRNA Transcriptome Involved in Bovine Intramuscular Fat Deposition.

BackgroundIntramuscular fat deposition in beef is a major determinant of carcass quality and value in the USA. The objective of this study was to examine changes in microRNA (miRNA) transcriptome that are involved with intramuscular fat deposition with time-on-concentrates (TOC). Yearling steers were individually fed a high concentrate diet and changes in intramuscular fat deposition were monitored by real-time ultrasound at 28 to 33 d intervals. Longissimus muscle biopsies collected on d 0, 92 and 124 TOC to examine changes in miRNA transcriptome that are involved in intramuscular fat deposition.ResultsSteer body weight increased (P < 0.0001) at each weigh day during TOC. Fat thickness increased (P < 0.005) from d 28 to 124. Ribeye area was larger (P < 0.001) on d 124 than d 61, which was larger than d 0 and 28. Ultrasound intramuscular fat content was greater (P < 0.001) on d 92 and 124 compared to d 0, 28 or 61. Sequencing of the muscle biopsy samples identified one miRNA, bta-miR-122, that was up-regulated (P < 0.005) at d 92 and 124 compared to d 0. At d 92 TOC, mRNA expression levels of fatty acid binding protein 4 (FABP4) and elongase 6 (ELOVL6) were up-regulated (P < 0.01) compared to d 0; whereas at d 124, lipogenic genes involved in de novo fatty acid synthesis, fatty acid transport, elongation and desaturation were highly up-regulated compared to d0.ConclusionsSmall RNA sequencing identified bta-miR-122 as a potential miRNA of interest that may be involved in intramuscular fat deposition with increasing TOC. Increased intramuscular fat content, as measured by real-time ultrasound, combined with differential gene expression suggests that preadipocyte differentiation may be stimulated first, which is followed by a global up-regulation of lipogenic genes involved in de novo fatty acid synthesis that provide fatty acids for subsequent hypertrophy.

Prognostic impact of insulin-like growth factor-I and its binding proteins, insulin-like growth factor-I binding protein-2 and -3, on adverse histopathological features and survival outcomes after radical cystectomy.

ObjectivesInsulin‐like growth factor‐I and its binding proteins are involved in cancer development, progression, and metastasis. In urothelial carcinoma, the impact of this pathway is still poorly investigated. The present large cohort study aimed to evaluate the association of preoperative circulating levels of insulin‐like growth factor‐I, insulin‐like growth factor‐I binding protein‐2 and ‐3 on outcomes after radical cystectomy.MethodsA retrospective cohort study of the plasma specimens from 1036 consecutive urothelial carcinoma patients who were treated with radical cystectomy. The primary and secondary outcomes were adverse histopathological features and survival outcomes. Binominal logistic regression and multivariable Cox regression analyses were performed to assess the association of plasma levels of insulin‐like growth factor‐I, insulin‐like growth factor‐I binding protein‐2 and ‐3 with outcomes.ResultsOn multivariable analysis adjusting for the effects of preoperative variables, lower insulin‐like growth factor‐I binding protein‐2 levels were associated with an increased risk of lymph node metastasis and (any non‐organ confined disease) any non‐organ confined disease. Insulin‐like growth factor‐I binding protein‐3 levels were also inversely independently associated with lymph node metastasis. Receiver operating characteristic curve analysis showed that the addition of insulin‐like growth factor‐I binding proteins biomarkers to a reference model significantly improved the discriminating ability for the prediction of lymph node metastasis (+10.0%, P < 0.001). On multivariable Cox regression models, lower levels of both insulin‐like growth factor‐I binding protein‐2 and ‐3 plasma levels were associated with recurrence‐free survival, cancer‐specific survival, and overall survival. insulin‐like growth factor‐I binding protein‐2 and ‐3 levels and improved the discrimination of a standard reference model for the prediction of recurrence‐free survival, cancer‐specific survival, and overall survival (+4.9%, 4.9%, 2.3%, respectively).ConclusionsPreoperative insulin‐like growth factor‐I binding protein‐2 and ‐3 are significantly associated with features of biologically and clinically aggressive urothelial carcinoma. These biomarkers improved prognostic urothelial carcinoma models.

Acute Administration of Metformin Protects Against Neuronal Apoptosis Induced by Cerebral Ischemia-Reperfusion Injury via Regulation of the AMPK/CREB/BDNF Pathway.

Metformin is a first-line anti-diabetic agent with a powerful hypoglycemic effect. Several studies have reported that metformin can improve the prognosis of stroke patients and that this effect is independent of its hypoglycemic effect; however, the specific mechanism remains unclear. In this research, we explored the effect and specific mechanism of metformin in cerebral ischemia-reperfusion (I/R) injury by constructing a transient middle cerebral artery occlusion model in vivo and a glucose and oxygen deprivation/reoxygenation (OGD/R) model in vitro. The results of the in vivo experiments showed that acute treatment with low-dose metformin (10 mg/kg) ameliorated cerebral edema, reduced the cerebral infarction volume, improved the neurological deficit score, and ameliorated neuronal apoptosis in the ischemic penumbra. Moreover, metformin up-regulated the brain-derived neurotrophic factor (BDNF) expression and increased phosphorylation levels of AMP-activated protein kinase (AMPK) and cAMP-response element binding protein (CREB) in the ischemia penumbra. Nevertheless, the above-mentioned effects of metformin were reversed by Compound C. The results of the in vitro experiments showed that low metformin concentrations (20 μM) could reduce apoptosis of human umbilical vein endothelial cells (HUVECs) under OGD/R conditions and promote cell proliferation. Moreover, metformin could further promote BDNF expression and release in HUVECs under OGD/R conditions via the AMPK/CREB pathway. The Transwell chamber assay showed that HUVECs treated with metformin could reduce apoptosis of SH-SY5Y cells under OGD/R conditions and this effect could be partially reversed by transfection of BDNF siRNA in HUVECs. In summary, our results suggest that metformin upregulates the level of BDNF in the cerebral ischemic penumbra via the AMPK/CREB pathway, thereby playing a protective effect in cerebral I/R injury.

Thioredoxin interacting protein protects mice from fasting induced liver steatosis by activating ER stress and its downstream signaling pathways

Under normal conditions, fasting results in decreased protein disulfide isomerase (PDI) activity and accumulation of unfolded proteins, leading to the subsequent activation of the unfolded protein response (UPR)/autophagy signaling pathway to eliminate damaged mitochondria. Fasting also induces upregulation of thioredoxin-interacting protein (TXNIP) expression and mice deficient of this protein (TXNIP-KO mice) was shown to develop severe hypoglycemia, hyperlipidemia and liver steatosis (LS). In the present study, we aimed to determine the role of TXNIP in fasting-induced LS by using male TXNIP-KO mice that developed LS without severe hypoglycemia. In TXNIP-KO mice, fasting induced severe microvesicular LS. Examinations by transmission electron microscopy revealed mitochondria with smaller size and deformities and the presence of few autophagosomes. The expression of β-oxidation-associated genes remained at the same level and the level of LC3-II was low. PDI activity level stayed at the original level and the levels of p-IRE1 and X-box binding protein 1 spliced form (sXBP1) were lower. Interestingly, treatment of TXNIP-KO mice with bacitracin, a PDI inhibitor, restored the level of LC3-II after fasting. These results suggest that TXNIP regulates PDI activity and subsequent activation of the UPR/autophagy pathway and plays a protective role in fasting-induced LS.

Abstract 91: ACTH stimulated salivary cortisol is a better indicator of adrenal insufficiency compared to stimulated serum cortisol in subjects with cirrhosis liver

Background: There are no reliable methods in clinical practice to diagnose adrenal insufficiency (AI) in patients with cirrhosis due to variably decreased cortisol binding protein levels. This leads to unreliable results on ACTH stimulated serum cortisol test.Aims and Objectives: To estimate the Long Acting (LA)-ACTH stimulated salivary cortisol levels of patients in different stages of cirrhosis using 2nd generation electrochemiluminescence and to find the prevalence of adrenal insufficiency in them.Results: LA-ACTH stimulated serum and saliva samples of 135 non-critically ill patients with cirrhosis (45 each from CHILD A, B and C) and 40 normal subjects were collected at baseline, 60 and 120-minutes. In healthy subjects, the 2.5th centile of 2 hr ACTH stimulated serum and salivary cortisol were 19.8 μg/dl and 0.97 μg/dl, which were used as cut-offs for defining AI based on serum and saliva respectively. The median (interquartile-range) 2-hour stimulated salivary cortisol in Child A,B,C categories and controls were 1.36 (1.23-2.38), 1.46 (1.18-2.22), 1.72 (1.2-2.2) and 2.12 (1.42-2.72) μg/dl respectively. 7 subjects (5.1%) were diagnosed to have AI based on stimulated salivary cortisol cut-off, whereas 39 (28.9%) cirrhosis subjects had inadequately stimulated serum cortisol. Three (symptomatic) subjects required steroid replacement.Conclusions: ACTH stimulated serum cortisol is a poor test to diagnose AI in patients with cirrhosis. ACTH stimulated salivary cortisol has a high reliability for the same.

Effect of moxibustion with wheat-grain-sized moxa cones on uterine natural killer cells and deci-dualization in rats with thin endometrium

To observe the effect of moxibustion at "Shenshu"(BL23) and "Guanyuan" (CV4) on decidua-lization and uterine natural killer cells in rats with thin endometrium, so as to explore its mechanism underlying promotion of embryo implantation. Female SD rats were randomly divided into normal control, model and wheat-grain-sized moxa cone moxibustion (moxibustion) groups, with 14 rats in each group. The thin endometrium model was established by bilaterally intrauterine perfusion of 95% ethanol (first) and saline (later) during estrus. For rats of the moxibustion group, the ignited wheat-grain-sized moxa cones were applied to bilateral BL23 and CV4, with 7 moxa cones for each acupoint, once a day, continuously for 3 estrous cycles. Then the male and female rats were raised in the same cage. On the 5th day of pregnancy, the rats were killed under anesthesia and the uterus tissue was collected for measuring the endometrium thickness and the numbers of blood vessels and glands after H.E. staining, detecting the levels of the proportion of natural killer cells with flow cytometry. After the uterine natural killer cells were sorted by the immunomagnetic bead method, the expression levels of insulin-like growth factor binding protein (IGFBP-1), interferon(INF-γ), tumor necrosis factor(TNF-α), transforming growth factor(TGF-β), interleukin 4(IL-4) and IL-10 mRNAs were detected by using fluorescence quantitative real-time PCR. Compared with the normal control group, the endometrium thickness, number of glands and blood vessels, and the expression levels of IGFBP-1, TGF-β, IL-4 and IL-10 mRNAs were significantly decreased (P<0.01, P<0.05), while the expression levels of IFN-γ and TNF-α mRNAs were significantly increased (P<0.05，P<0.01) in the model group. In contrast to the model group, the endometrium thickness, number of glands and blood vessels, and the expression levels of IGFBP-1, TGF-β, IL-4 and IL-10 mRNAs were significantly increased (P<0.05, P<0.01), while the expression levels of IFN-γ and TNF-α mRNAs were considerably down-regulated (P<0.05, P<0.01) in the moxibustion group. No significant difference was found among the 3 groups in the proportion of natural killer cells in the endometrium (P>0.05). Moxibustion of BL23 and CV4 with wheat-grain-sized moxa cones can improve the degree of thin endometrial decidualization, which may be related with its functions in regulating the levels of cytokines secreted from natural killer cells in the uterus.

Association between Serum Adipocyte Fatty Acid Binding Protein Level and Endothelial Dysfunction in Chronic Hemodialysis Patients.

Adipocyte fatty acid binding protein (A-FABP) is associated with atherosclerosis, and endothelial dysfunction is one of the reasons for adverse cardiovascular outcomes in patients undergoing hemodialysis (HD). This study investigated the correlation between serum A-FABP levels and endothelial function in HD patients. Fasting blood samples were collected from 90 HD patients. A-FABP levels were measured using a commercial enzyme immunoassay kit. Endothelial function was evaluated by a digital thermal monitoring test to measure vascular reactivity index (VRI). VRI < 1.0, 1.0 ≤ VRI < 2.0, and VRI ≥ 2.0 indicated poor, intermediate, and good vascular reactivity, respectively. In total, 14 (15.6%), 38 (42.2%), and 38 (42.2%) HD patients had poor, intermediate, and good VRI, respectively. Patients with poor VRI had lower pre-HD and post-HD body weight, body mass index, and serum creatinine level but higher serum A-FABP level (p = 0.001) than those with intermediate and good VRI. Log-transformed VRI (log-VRI) positively correlated with serum creatinine and negatively correlated with A-FABP by multivariate linear regression analysis. We concluded that A-FABP correlated with endothelial dysfunction in chronic HD patients.

Endothelin-A Receptor Antagonist Alleviates Allergic Airway Inflammation via the Inhibition of ILC2 Function.

Allergic airway inflammation is a universal airway disease that is driven by hyperresponsiveness to inhaled allergens. Group 2 innate lymphoid cells (ILC2s) produce copious amounts of type 2 cytokines, which lead to allergic airway inflammation. Here, we discovered that both peripheral blood of human and mouse lung ILC2s express the endothelin-A receptor (ETAR), and the expression level of ETAR was dramatically induced upon interleukin-33 (IL-33) treatment. Subsequently, both preventive and therapeutic effects of BQ123, an ETAR antagonist, on allergic airway inflammation were observed, which were associated with decreased proliferation and type 2 cytokine productions by ILC2s. Furthermore, ILC2s from BQ123 treatment were found to be functionally impaired in response to an interleukin IL-33 challenged. And BQ123 treatment also affected the phosphorylation level of the extracellular signal-regulated kinase (ERK), as well as the level of GATA binding protein 3 (GATA3) in activated ILC2s. Interestingly, after BQ123 treatment, both mouse and human ILC2s in vitro exhibited decreased function and downregulation of ERK signaling and GATA3 stability. These observations imply that ETAR is an important regulator of ILC2 function and may be involved in ILC2-driven pulmonary inflammation. Therefore, blocking ETAR may be a promising therapeutic strategy for allergic airway inflammation.

Activity of Protein S-C4b Binding Protein and Total TFPI Levels in Egyptian SLE Patients: A Cross-Sectional Study

BACKGROUND: Systemic lupus erythematosus (SLE) is an immune disorder with alternating active and remission phases. Cardiovascular diseases and thrombosis are the major causes of mortality in SLE. The anticoagulant activity of Protein S (PS) is complemented by C4 binding protein (C4BP) and tissue factor pathway inhibitor (TFPI). AIM: This study aims to determine the extent of change in the levels of PS activity, C4BP, and total TFPI in active SLE in comparison to the SLE remission phase and their association with thrombosis during SLE flare. METHODS: The study included 180 Egyptian SLE patients who were classified into two groups: 100 SLE cases as the active group and 80 SLE cases as the remission group. The PS activity levels were processed on automated coagulation analyzers, whereas the C4BP and total TFPI levels were measured via enzyme-linked immunosorbent assay. RESULTS: The PS activity and C4BP levels were lower in the active SLE cases than in the remitted ones (p &lt; 0.05). The levels of PS activity and C4BP were revealed to be independent predictors of SELENA-SLEDAI flare scores. In active SLE cases, the PS activity and C4BP levels were rated as excellent and fair classifiers of thrombotic risk in SLE flare, respectively. The total TFPI levels showed no association with SLE activity or its thrombotic consequences. CONCLUSIONS: The levels of PS activity and C4BP act as important biomarkers for SLE activity. Both can be implanted as predictive tools for thrombosis during activity.

Clinical study on the minimally invasive percutaneous nephrolithotomy treatment of upper urinary calculi.

BACKGROUNDUpper urinary tract stones are very common in my country, with an incidence of 1% to 5% in the North and an even higher incidence of 5% to 10% in the south. The incidence rate in the south is higher than that in the north, mainly due to the water quality, climate and eating habits of the region. From the perspective of sex, incidence is more likely in males than females. In the high-incidence population, young adults are most prone to stones. Men in the age range of 25 to 40 years are more likely to have stones.AIMTo observe the therapeutic effect of minimally invasive percutaneous nephrolithotomy (mPCNL) on upper urinary tract stones and its influence on the renal function of patients.METHODSPatients with upper urinary tract stones who were treated in our hospital from February 2017 to March 2018 were selected as research subjects and were divided into the PCNL group and the mPCNL group according to the random number table method. The general conditions of the two groups of patients were observed during the perioperative period, and the differences in stone clearance, pain, renal function indicators and complication rates were compared between the two groups to determine which were statistically significant (P < 0.05).RESULTSThe operation time of the mPCNL group was longer than that of the PCNL group (t = -34.392, P < 0.001), and the intraoperative blood loss of the mPCNL group was more than that of the PCNL group (t = 34.090, P < 0.001). There was no difference in renal function indices between the two groups of patients before treatment, and there was no difference in the levels of serum creatinine, β2 microglobulin or retinol binding protein in the mPCNL group after treatment. The visual analog scale score of patients in the mPCNL group was lower than that of the PCNL group (t = 12.191, P < 0.001), and there was no significant difference in the stone clearance rate between the two groups (χ2 value = 1.013, P = 0.314). There was no significant difference in the incidence of urine extravasation, dyspnea and peripheral organ damage between the two groups (χ2 value = 1.053, P = 0.305). At 1 mo after treatment and 3 mo after treatment, the quality of life of the mPCNL group was lower than that of the PCNL group, and the Qmax level of the mPCNL group was higher than that of the PCNL group.CONCLUSIONmPCNL has a good therapeutic effect on upper urinary tract stones, with a high stone clearance rate without causing kidney damage or increasing the incidence of complications, and thus has good application value.