NE Levels Research Articles

Poliumoside inhibits apoptosis, oxidative stress and neuro-inflammation to prevent intracerebroventricular Streptozotocin-induced cognitive dysfunction in Sprague-Dawley Rats: in in-vivo, in-vitro and in-silico study.

Alzheimer's disease (AD) is a severe neurological illness causes cognitive decline and mortality if not treated early. However, the current therapeutic modalities are inefficient to manage the cognitive dysfunction of AD. Therefore, in the present manuscript, we have enumerated the pharmacological benefit of Poliumoside in the Streptozotocin-induced cognitive dysfunction in Sprague-Dawley (SD) rats. Initially, the cognitive dysfunction in rats was induced by the intracerebroventricular administration of Streptozotocin, then rats received PMD (5 mg and 10 mg/kg body weight) was given. Various behavioural analysis, such as Morris water maze (MWM), and object recognition tests (ORT), and locomotor analysis was conducted in PMD treated group. Various biochemical analysis was conducted to analyze the effect of PMD on hippocampus oxidative-nitrosative stress and pro-inflammatory cytokines. MTT assay and annexin V/PI staining was performed to analyse the effect of PMD on the cell viability and neuronal toxicity of PC12 cells, respectively. Molecular docking analysis was also conducted with crystal structure of human AChE. PMD treatment improved cognitive capacity in rats in MWM and ORT. Compared to STZ rats, PMD-treated rats had significantly higher locomotor activity and lower AChE activity. PMD also restores dopamine, 5-HT, and NE levels and reduces metabolic their deactivation as evidenced by increased levels of DOPAC, HVA, 5-HIAA. Nitrite, MDA, SOD, CAT, and GSH levels were restored near normal in PMD-treated rats, reducing hippocampus oxidative-nitrosative stress. Pro-inflammatory cytokines were similarly lowered in PMD-treated rats. In in-vitro studies, PMD did not affect PC12 cell survival at the maximal dose of 10 µM. In addition, PMD concentration-dependently prevents H₂O₂-induced neuronal death in PC12 cells. The in-silico docking analysis showed that the PMD fit snugly into the active site of human AChE by engaging with the anionic domain and the catalytic triad of Trp86, Tyr337, Phe338, and Gly121 residues. In conclusion, our study demonstrated that PMD have significant impact on AD by inhibiting ACheE and restoring neurotransmitter levels, which enhances Ach levels in rats and improves cognitive impairment in STZ rats.

Analysis of Clinical Characteristics of Insomnia Patients and Effect of Acupuncture Therapy

Objective: To explore the clinical characteristics of insomnia patients and the function and mechanism of acupuncture therapy, to improve the recognition and diagnosis of insomnia. Methods: The clinical data of 60 patients with insomnia who met the criteria and were admitted to the Department of Acupuncture and moxibustion in Yunnan Provincial Hospital of Integrated Traditional Chinese and Western Medicine from January 2023 to February 2024 were selected, the clinical characteristics of the patients were analyzed, and the patients were randomly divided into two groups for routine treatment and acupuncture treatment, respectively, to explore the therapeutic effect and possible mechanism. Results: The insomnia symptoms of the patients were mainly difficulty in falling asleep. Acupuncture treatment could significantly improve the insomnia symptoms and promote the expression levels of DA and NE. Conclusion: Patients with insomnia have various clinical features, mainly sleep difficulties. Acupuncture treatment can significantly relieve insomnia symptoms by up-regulating DA and NE levels, and then improve sleep quality.

ERβ activation improves nonylphenol-induced depression and neurotransmitter secretion disruption via the TPH2/5-HT pathway

The aim of this study is to investigate the role of estrogen receptor β (ERβ) in nonylphenol (NP) - induced depression - like behavior in rats and its impact on the regulation of the TPH2/5-HT pathway. In the in vitro experiment, rat basophilic leukaemia cells (RBL-2H3) cells were divided into the four groups: blank group, NP group (20 μM), ERβ agonist group (0.01 μM), and NP＋ERβ agonist group (20 μM＋0.01 μM). For the in vivo experiment, 72 adult male Sprague-Dawley rats were randomly divided into following six groups: the Control, NP (40 mg/kg) group, ERβ agonist (2 mg/kg, Diarylpropionitrile (DPN)) group, ERβ inhibitor (0.1 mg/kg, 4-(2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl) phenol (PHTPP)) group, NP+ERβ agonist (40 mg/kg NP ＋ 2 mg/kg DPN) group, and NP+ERβ inhibitor (40 mg/kg NP + 0.1 mg/kg PHTPP) group, with 12 rats in each group. Each rat in drug group were given NP by gavage and/or received a single intraperitoneal injection of DPN 2 mg/kg or PHTPP 0.1 mg/kg. Both in vivo and in vitro, NP group showed a decrease in the expression levels of ERβ, tryptophan hydroxylase (TPH1), and tryptophan hydroxylase-2 (TPH2) genes and proteins, and reduced levels of DA, NE, and 5-hydroxytryptophan (5-HT) neurotransmitters. RBL-2H3 cells showed signs of cell shrinkage, with rounded cells, increased suspension and more loosely arranged cells. The effectiveness of the ERβ agonist stimulation exhibited an increase exceeding 60% in RBL-2H3 cells. The application of ERβ agonist resulted in an alleviation the aforementioned alterations. ERβ agonist activated the TPH2/5-HT signaling pathways. Compared to the control group, the NP content in the brain tissue of the NP group was significantly increased. The latency to eat for the rats was longer and the amount of food consumed was lower, and the rats had prolonged immobility time in the behavioral experiment of rats. The expression levels of ERβ, TPH1, TPH2, 5-HT and 5-HITT proteins were decreased in the NP group, suggesting NP-induced depression-like behaviours as well as disturbances in the secretion of serum hormones and monoamine neurotransmitters. In the NP group, the midline raphe nucleus showed an elongated nucleus with a dark purplish-blue colour, nuclear atrophy, displacement and pale cytoplasm. ERβ might ameliorate NP-induced depression-like behaviors, and secretion disorders of serum hormones and monoamine neurotransmitters via activating TPH2/5-HT signaling pathways.

Repetitive mild traumatic brain injury impairs norepinephrine system function and psychostimulant responsivity

Traumatic brain injury (TBI) is a complex pathophysiological process that results in a variety of neurotransmitter, behavioral, and cognitive deficits. The locus coeruleus-norepinephrine (LC-NE) system is a critical regulator of arousal levels and higher executive processes affected by TBI including attention, working memory, and decision making. LC-NE axon injury and impaired signaling within the prefrontal cortex (PFC) is a potential contributor to the neuropsychiatric symptoms after single, moderate to severe TBI. The majority of TBIs are mild, yet long-term cognitive deficits and increased susceptibility for further injury can accumulate after each repetitive mild TBI. As a potential treatment for restoring cognitive function and daytime sleepiness after injury psychostimulants, including methylphenidate (MPH) that increase levels of NE within the PFC, are being prescribed “off-label”. The impact of mild and repetitive mild TBI on the LC-NE system remains limited. Therefore, we determined the extent of LC-NE and arousal dysfunction and response to therapeutic doses of MPH in rats following experimentally induced single and repetitive mild TBI. Microdialysis measures of basal NE efflux from the medial PFC and arousal measures were significantly lower after repetitive mild TBI. Females showed higher baseline PFC-NE efflux than males following single and repetitive mild TBI. In response to MPH challenge, males exhibited a blunted PFC-NE response and persistent arousal levels following repetitive mild TBI. These results provide critical insight into the role of catecholamine system dysfunction associated with cognitive deficits following repeated injury, outcome differences between sex/gender, and lack of success of MPH as an adjunctive therapy to improve cognitive function following injury.

Revealing functional-molecular parasympathetic dysfunction in PVC-induced cardiomyopathy via dual stressor PVC and exercise

Abstract Background Exercise intolerance is a common presentation of premature ventricular contractions-induced cardiomyopathy (PVC-CM). In a canine PVC-CM model, previous studies have shown cardiac autonomic nervous system remodeling, resulting in persistent sympathetic excess. Nevertheless, the molecular mechanisms and functional implications of this neural remodeling, as well as parasympathetic alterations, remain unclear. Purpose We aim to evaluate the functional-molecular mechanisms of parasympathetic remodeling and its potential contribution to exercise intolerance in an animal model of PVC-CM. Methods We studied 15 canines (8 experimental vs 7 sham controls) chronically instrumented with pacemakers and telemetry devices to deliver 12-weeks of bigeminal PVCs (200ms coupling) to induce PVC-CM and to record sympathetic (SNA), vagal nerve activity (VNA) and heart rate (HR) over this period. Exercise challenge was conducted in sinus rhythm (SR) and in the presence of bigeminal PVCs ("dual stressor") at baseline and after development of PVC-CM. Sympatho-vagal neural and HR responses were assessed during and after exercise at baseline and after PVC-CM development, with and without PVCs, thus comparing baseline SR, baseline with PVCs, CM SR and CM with PVCs. Western Blot was performed on intrinsic and extrinsic cardiac neural tissues to evaluate molecular mechanisms of neural remodeling. Results Compared to baseline, SNA was significantly higher in PVC-CM in all phases of exercise (rest, exercise, recovery, p=0.03). In contrast, VNA was significantly lower in PVC-CM than baseline during exercise recovery (P=0.014). After exercise, HR recovery (HRR) was impaired (p=0.001), VNA lower (p=0.014) and SNA (p=0.03) higher in PVC-CM compared with baseline. Tyrosine hydroxylase (TH) immunoreactivity in the left ventricle (LV, p=0.001) and stellate ganglia (SG, p=0.03) and trans-cardiac plasma NE levels (p=0.03) were higher in PVC-CM compared with sham. Choline Acetyltransferase (CHAT) was not significantly different in either vagus (p=0.33) or LV (p=0.8) between PVC-CM vs sham. Cardiac (LV) Nerve Growth Factor (NGF), Growth Associated Protein (GAP-43) and beta 1-adrenoreceptor levels were lower (p=0.042, p=0.003, p=0.002 respectively) whereas beta 2-adreno- and muscarinic M2-cholinergic receptors were higher (p=0.02, p<0.01 respectively) in PVC-CM compared with sham. Conclusion Neural remodeling in PVC-CM results in an imbalanced sympathetic excess and sympathetic-parasympathetic dysfunction characterized by a loss of sympathetic functional reserve during exercise and impaired parasympathetic activity and HRR after exercise. Adaptive downregulation of NGF might explain this imbalance and dysfunction. These data open the intriguing possibility of neural-targeted therapies to reverse remodeling and restore autonomic function disrupted by chronic PVCs.GraphsMolecular

Protective role of TRPV2 in synaptic plasticity through the ERK1/2-CREB-BDNF pathway in chronic unpredictable mild stress rats

PurposeChronic stress is a significant risk factor for mood disorders such as depression, where synaptic plasticity plays a central role in pathogenesis. Transient Receptor Potential Vanilloid Type-2 (TRPV2) Ion Channels are implicated in hypothalamic-pituitary-adrenal axis disorders. Previous proteomic analysis indicated a reduction in TRPV2 levels in the chronic unpredictable mild stress (CUMS) rat model, yet its role in synaptic plasticity during depression remains to be elucidated. This study aims to investigate TRPV2's role in depression and its underlying mechanisms. MethodsIn vivo and in vitro experiments were conducted using the TRPV2-specific agonist probenecid and ERK1/2 inhibitors SCH772984. In vivo, rats underwent six weeks of CUMS before probenecid administration. Depressive-like behaviors were assessed through behavioral tests. ELISA kits measured 5-HT, DA, NE levels in rat hippocampal tissues. Hippocampal morphology was examined via Nissl staining. In vitro, rat hippocampal neuron cell lines were treated with ERK1/2 inhibitors SCH772984 and probenecid. Western blot, immunofluorescence, immunohistochemical staining, and RT-qPCR assessed TRPV2 expression, neurogenesis-related proteins, synaptic markers, and ERK1/2-CREB-BDNF signaling proteins. ResultsDecreased hippocampal TRPV2 levels were observed in CUMS rats. Probenecid treatment mitigated depressive-like behavior and enhanced hippocampal 5-HT, NE, and DA levels in CUMS rats. TRPV2 activation countered CUMS-induced synaptic plasticity inhibition. Probenecid activated the ERK1/2-CREB-BDNF pathway, suggesting TRPV2's involvement in this pathway via ERK1/2. ConclusionThese findings indicate that TRPV2 activation offers protective effects against depressive-like behaviors and enhances hippocampal synaptic plasticity in CUMS rats via the ERK1/2-CREB-BDNF pathway. TRPV2 emerges as a potential therapeutic target for depression.

143 Effect of increasing dietary net energy content at a constant lysine on feed intake and performance of 7 to 11 kg and 11 to 20 kg pigs

Abstract Information about NE requirements for nursery pigs is still limited. Therefore, 2 experiments were conducted to evaluate the effect of dietary NE contents on growth performance of 7 to 11 kg pigs (Exp. 1) and 11 to 20 kg pigs (Exp. 2). In Exp. 1, mixed-sex pigs (n = 192; PIC; initial body weight (BW) of 7.8 ± 1.07 kg] were assigned to 4 diets for 14 d. Diets 1 to 4 were formulated to contain 4 levels of NE (9.8, 10.1, 10.4 and 10.7 MJ/kg) and adequate in all AA with standardized ileal digestible (SID) Lys being constant at 1.42%. In Exp. 2, mixed-sex pigs (n = 192; PIC; initial BW of 11.2 ± 1.54 kg) were assigned to 4 diets for 21 d. Diets 1 to 4 contained 4 levels of NE (9.6, 9.9, 10.2 and 10.5 MJ/kg) and adequate in all AA (1.32% SID Lys). For both studies, there were 6 pen replicates (4 entire males and 4 gilts/pen) per treatment, and diets were based on corn, soybean meal, whey powder, wheat bran and soy oil. Pigs had ad libitum access to feed (mash) and water. Individual pigs and pen feed disappearance were weighed weekly to calculate average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency gain to feed ratio (G:F). Data were analyzed by ANOVA using the GLM procedure of SAS with pen as the experimental unit. In Exp. 1, ADFI tended to decrease (quadratic; P ≤ 0.08) with increasing NE levels during d 0 to 7 and d 8 to 14. The ADG during d 0 to 7 tended to be greater (158, 192, 187 and 152 g for diets 1 to 4; quadratic; P = 0.08) at 10.1 MJ/kg NE; however, the overall ADG (d 0 to 14) was not affected by the NE levels. Although not significant, G:F, Lys utilization efficiency were greatest at 10.1 MJ/kg NE for the overall period. In Exp. 2, ADFI decreased (827, 783, 768 and 749 g for diets 1 to 4; linear; P = 0.05) with increasing NE levels during d 15 to 21 but was not affected (P > 0.05) for other periods. The ADG (d 8 to 14) tended to be greater at 9.6 MJ/kg NE (quadratic; P = 0.06) and G:F (d 15 to 21) tended to be greater at a dietary NE of 10.2 MJ/kg (linear; P = 0.10). However, the overall (d 0 to 21) ADG and G:F were not affected (P > 0.05) by NE levels. Although not significant, G:F and Lys utilization efficiency were greatest at a dietary NE of 10.2 MJ/kg for the overall period. These results indicate that G:F of 7 to 11 kg pigs and 11 to 20 kg pigs maximized at a dietary NE content of 10.1 MJ/kg and 10.2 MJ/kg, which correspond to 1.41 and 1.29 g SID Lys/MJ NE, respectively.

291 An evaluation of soybean meal inclusion rate in pigs fed from wean to market

Abstract Soybean meal (SBM) supply may result in decreased relative costs of SBM compared with other ingredients. Net energy of SBM closer to the NE of corn has been reported which may increase its relative value in formulations (Boyd and Gaines, 2023). To evaluate the effect of SBM level in two common diet formulation strategies, Corn-DDGS-SBM (DDGS) or Corn-SBM (SBM) in wean-to-market pigs (WTM). A total of 540 pigs (PIC X Duroc, 18 d of age, 5.3 kg) were weaned and blocked by body weight (BW) and sex for allocation. Six dietary treatments were assigned to 90 pens to give 15 replications per treatment in a randomized complete block design (RCBD). Individual BW for average daily gain (ADG) and pen feed disappearance (ADF) were collected every 2 wk for 18 wk and carcass data were collected at average market weight of 129 kg (18 to 20 wk after placement). The mash diets were formulated to a SID lysine:NE target in 7 phases using current NE estimates for SBM, 2087 Kcal/kg, and to meet or exceed NRC requirements (2012). The NE level used for DDGS was 2,343 Kcal/kg. The treatments consisted of Low (LO), Medium (MED), and High (HI) SBM levels by providing 0.5%, 0.25% and no added lysine (AA) in SBM- or DDGS (30%)-based diets. Data were analyzed using pen as the experimental unit for performance data and the pig as the experimental unit for carcass data. Data were analyzed using the MIXED procedure (SAS Inst. Inc., Cary, NC) and P < 0.05 was considered significant and 0.05 ≤ P < 0.10 considered a tendency. The ADF tended to be less from wk 0 to 18 for pigs fed SBM than DDGS diets. SBM diets increased wk 6 BW and ADG compared with DDGS (P < 0.05). HI and MED SBM additions increased wk 6 BW and wk 0 to 6 ADG vs LO SBM (P < 0.05). SBM diets increased wk 14 and 18 BW, gain to feed ratio (G:F) and ADG compared with DDGS (P < 0.05) except for MED in DDGS diets, which was not different. HI and MED SBM additions increased wk 14 and 18 BW and 0 to 18 wk ADG vs LO SBM (P < 0.05). Carcass data indicated SBM diets increased hot carcass weight by 2 kg, loin depth, back fat and calculated lean weight, compared with DDGS (P < 0.05). LO SBM resulted in more back fat, less loin depth and lean weight compared with the use of HI or MED SBM (P < 0.05). SBM improved growth, feed and caloric conversion, and increased carcass value in high-performance WTM pigs compared with the use of DDGS. MED and HI SBM level tended to improve ADG and carcass characteristics.

24 Effects of reducing dietary net energy on growth performance and carcass characteristics of grow-finish pigs

Abstract This study was conducted to evaluate the effects of dietary NE on growth performance, carcass characteristics, and removal and mortality rates of grow-finish pigs. A total of 1,927 pigs [initial body weight (BW) 23.3 ± 0.23 kg] were used in a 125-d study. Pens of pigs were blocked by BW and randomly allotted to dietary treatments in a randomized complete block design with 23 to 26 pigs/pen and 15 replications/treatment. Pens received 1 of 5 diets with decreasing NE based on Eq. 1-8 NRC (2012) using proximate analysis of the major ingredients. A corn-soybean meal-based control diet was blended with a low energy diet that contained 8% less NE through inclusion of 25% wheat middlings and 15% DDGS (6% oil) to achieve intermediate NE levels. Thus, dietary NE fed through 5 dietary phases were: phase 1 (2,432, 2,383, 2,335, 2,286, and 2,237 kcal/kg); phase 2 (2,463, 2,413, 2,363, 2,315, and 2,266 kcal/kg); phase 3 (2,495, 2,445, 2,395, 2,345, and 2,295); phase 4 (2,518, 2,468, 2,417, 2,367, and 2,317 kcal/kg); and phase 5 (2,537, 2,486, 2,435, 2,384, 2,334 kcal/kg). Dietary phases were fed from approximately 23-40, 40-60, 60-80, 80-105, and 105-133 kg BW, respectively. Overall, reducing NE decreased (linear, P < 0.001) average daily gain (ADG) and final BW and increased (linear, P < 0.001) average daily feed intake (ADFI), resulting in decreased (linear, P < 0.001) gain to feed ratio (G:F; Table). Decreasing NE also tended to increase (linear, P = 0.073) the percentage of removals, with no evidence of difference (P > 0.10) in mortality. Reducing NE worsened (linear, P < 0.001) caloric efficiency (CE) based on NRC nutrient loading values, but improved (linear, P ≤ 0.003) CE based on INRA (2008) or Brazilian Tables for Poultry and Swine (2017) nutrient loading values. When formulating diets using CVB (2021) nutrient loading values, decreasing NE did not influence CE (P > 0.10). For carcass characteristics, decreasing NE decreased (linear, P < 0.001) HCW, backfat depth, and carcass yield. However, decreasing NE increased (linear, P ≤ 0.024) loin depth and percentage lean. When considering CE on a carcass gain basis, decreasing NE worsened (linear, P ≤ 0.004) CE based on NRC or CVB nutrient loading values, but improved (linear, P < 0.001) CE based on INRA nutrient loading values. Decreasing NE resulted in no differences (P > 0.010) in CE based on Brazilian Tables nutrient loading values. These results suggest that the NRC database overestimates NE contributions of fibrous ingredients, such as wheat middlings and corn DDGS, while the INRA database underestimates their contributions to NE, whether CE is calculated based on live or carcass gain. Meanwhile, NE values in the CVB database appear to assess nutrient values more accurately for NE based on live gain but underestimate their value on a carcass basis. The Brazilian tables appear to underestimate the contributions of fibrous ingredients to dietary NE on a live basis, but more accurately value their contributions on a carcass basis.

Excessive fat expenditure in MCT-induced heart failure rats is associated with BMAL1/REV-ERBα circadian rhythmic loop disruption

Fat loss predicts adverse outcomes in advanced heart failure (HF). Disrupted circadian clocks are a primary cause of lipid metabolic issues, but it's unclear if this disruption affects fat expenditure in HF. To address this issue, we investigated the effects of disruption of the BMAL1/REV-ERBα circadian rhythmic loop on adipose tissue metabolism in HF.50 Wistar rats were initially divided into control (n = 10) and model (n = 40) groups. The model rats were induced with HF via monocrotaline (MCT) injections, while the control group received equivalent solvent injections. After establishing the HF model, the model group was further subdivided into four groups: normal rhythm (LD), inverted rhythm (DL), lentivirus vector carrying Bmal1 short hairpin RNA (LV-Bmal1 shRNA), and empty lentivirus vector control (LV-Control shRNA) groups, each with 10 rats. The DL subgroup was exposed to a reversed light–dark cycle of 8 h: 16 h (dark: light), while the rest adhered to normal light–dark conditions (light: dark 12 h: 12 h). Histological analyses were conducted using H&E, Oil Red O, and Picrosirius red stains to examine adipose and liver tissues. Immunohistochemical staining, RT-qPCR, and Western blotting were performed to detect markers of lipolysis, lipogenesis, and beiging of white adipose tissue (WAT), while thermogenesis indicators were detected in brown adipose tissue (BAT). The LD group rats exhibited decreased levels of BMAL1 protein, increased levels of REV-ERBα protein, and disrupted circadian circuits in adipose tissue compared to controls. Additionally, HF rats showed reduced adipose mass and increased ectopic lipid deposition, along with smaller adipocytes containing lower lipid content and fibrotic adipose tissue. In the LD group WAT, expression of ATGL, HSL, PKA, and p-PKA proteins increased, alongside elevated mRNA levels of lipase genes (Hsl, Atgl, Peripilin) and FFA β-oxidation genes (Cpt1, acyl-CoA). Conversely, lipogenic gene expression (Scd1, Fas, Mgat, Dgat2) decreased, while beige adipocyte markers (Cd137, Tbx-1, Ucp-1, Zic-1) and UCP-1 protein expression increased. In BAT, HF rats exhibited elevated levels of PKA, p-PKA, and UCP-1 proteins, along with increased expression of thermogenic genes (Ucp-1, Pparγ, Pgc-1α) and lipid transportation genes (Cd36, Fatp-1, Cpt-1). Plasma NT-proBNP levels were higher in LD rats, accompanied by elevated NE and IL-6 levels in adipose tissue. Remarkably, morphologically, the adipocytes in the DL and LV-Bmal1 shRNA groups showed reduced size and lower lipid content, while lipid deposition in the liver was more pronounced in these groups compared to the LD group. At the gene/protein level, the BMAL1/REV-ERBα circadian loop exhibited severe disruption in LV-Bmal1 shRNA rats compared to LD rats. Additionally, there was increased expression of lipase genes, FFA β oxidation genes, and beige adipocyte markers in WAT, as well as higher expression of thermogenic genes and lipid transportation genes in BAT. Furthermore, plasma NT-proBNP levels and adipose tissue levels of NE and IL-6 were elevated in LV-Bmal1 shRNA rats compared with LD rats. The present study demonstrates that disruption of the BMAL1/REV-ERBα circadian rhythmic loop is associated with fat expenditure in HF. This result suggests that restoring circadian rhythms in adipose tissue may help counteract disorders of adipose metabolism and reduce fat loss in HF.

Homotherapy-for-heteropathy of Bupleurum Chinense DC.-Scutellaria baicalensis Georgi in treating depression and colorectal cancer: A network pharmacology and animal model approach

Ethnopharmacological relevanceBupleurum chinense DC.-Scutellaria baicalensis Georgi (BS) is a classic drug pair that has good clinical effects on depression and many tumors. However, the concurrent targeting mechanism of how the aforementioned drug pair is valid in the two distinct diseases, has not been clarified yet. Aim of the studyThe components of BS were detected by LC-MS, combined with network pharmacology to explore the active ingredients and common targeting mechanism of its multi-pathway regulation of BS in treating depression and CRC, and to validate the dual effects of BS using the CUMS mice model and orthotopic transplantation tumor mice model of CRC. ResultsTwenty-nine components were screened, 84 common gene targets were obteined, and the top 5 key targets including STAT3, PIK3R1, PIK3CA, AKT1, IL-6 were identified by PPI network. GO and KEGG analyses revealed that PI3K/AKT and JAK/STAT signaling pathways might play a crucial role of BS in regulating depression and CRC. BS significantly modulated CUMS-induced depressive-like behavior, attenuated neuronal damage, and reduced serum EPI and NE levels in CUMS model mice. BS improved the pathological histological changes of solid tumors and liver tissues and inhibited solid tumors and liver metastases in tumor-bearing mice. BS significantly decreased the proteins' expression of IL-6, p-JAK2, p-STAT3, p-PI3K, p-AKT1 in hippocampal tissues and solid tumors, and regulated the levels of IL-2, IL-6 and IL-10 in serum of two models of mice. ConclusionBS can exert dual antidepressant and anti-CRC effects by inhibiting the expression of IL-6/JAK2/STAT3 and PI3K/AKT pathway proteins and regulating the release of inflammatory cytokines.

Sedative and hypnotic effects of the saponins from a traditional edible plant Liriope spicata Lour. in PCPA-induced insomnia mice

Ethnopharmacological relevanceLiriope spicata Lour., a species listed in the catalogue of ‘Medicinal and Edible Homologous Species’, is traditionally used for the treatment of fatigue, restlessness, insomnia and constipation. Aim of the studyThis study is aimed to evaluate the sedative and hypnotic effect of the saponins from a natural plant L. spicata Lour. in vivo. Materials and methodsThe total saponin (LSTS) and purified saponin (LSPS) were extracted from L. spicata, followed by a thorough analysis of their major components using the HPLC-MS. Subsequently, the therapeutic efficacy of LSTS and LSPS was evaluated by the improvement of anxiety and depression behaviors of the PCPA-induced mice. ResultsLSTS and LSPS exhibited similar saponin compositions but differ in their composition ratios, with liriopesides-type saponins accounting for a larger proportion in LSTS. Studies demonstrated that both LSTS and LSPS can extend sleep duration and immobility time, while reducing sleep latency in PCPA-induced mice. However, there was no significant difference in weight change among the various mice groups. Elisa results indicated that the LSTS and LSPS could decrease levels of NE, DA, IL-6, and elevate the levels of 5-HT, NO, PGD2 and TNF-α in mice plasma. LSTS enhanced the expression of neurotransmitter receptors, while LSPS exhibited a more pronounced effect in regulating the expression of inflammatory factors. In conclusion, the saponins derived from L. spicata might hold promise as ingredients for developing health foods with sedative and hypnotic effects, potentially related to the modulation of serotonergic and GABAAergic neuron expression, as well as immunomodulatory process.

Increased Elastase and Matrix Metalloproteinase Levels in the Pulmonary Arteries of Infants With Congenital Diaphragmatic Hernia

BackgroundPulmonary vascular disease (PVD) complicated with pulmonary hypertension (PH) is a leading cause of mortality in congenital diaphragmatic hernia (CDH). Unfortunately, CDH patients are often resistant to PH therapy. Using the nitrogen CDH rat model, we previously demonstrated that CDH-associated PVD involves an induction of elastase and matrix metalloproteinase (MMP) activities, increased osteopontin and epidermal growth factor (EGF) levels, and enhanced smooth muscle cell (SMC) proliferation. Here, we aimed to determine whether the levels of the key members of this proteinase-induced pathway are also elevated in the pulmonary arteries (PAs) of CDH patients. MethodsNeutrophil elastase (NE), matrix metalloproteinase-2 (MMP-2), epidermal growth factor (EGF), tenascin-C, and osteopontin levels were assessed by immunohistochemistry in the PAs from the lungs of 11 CDH patients and 5 normal age-matched controls. Markers of proliferation (proliferating cell nuclear antigen (PCNA)) and apoptosis (cleaved (active) caspase-3) were also used. ResultsWhile expressed by both control and CDH lungs, the levels of NE, MMP-2, EGF, as well as tenascin-C and osteopontin were significantly increased in the PAs from CDH patients. The percentage of PCNA-positive PA SMCs were also enhanced, while those positive for caspase-3 were slightly decreased. ConclusionsThese results suggest that increased elastase and MMPs, together with elevated tenascin-C and osteopontin levels in an EGF-rich environment may contribute to the PVD in CDH infants. The next step of this study is to expand our analysis to a larger cohort, and determine the potential of targeting this pathway for the treatment of CDH-associated PVD and PH. Type of StudyTherapeutic. Level of EvidenceLEVEL III.

Higher pupillary dilation during phasic transcutaneous vagus nerve stimulation (taVNS) and its relevance to the noradrenergic system of the locus coeruleus (LC‐NE)

AbstractBackgroundTranscutaneous auricular vagus nerve stimulation (taVNS) may affect the locus coeruleus‐norepinephrine system (LC‐NE system) via modulation of the vagus nerve. The LC is our main source of NE in the brainstem and declines with age and is a potential epicentre of tau pathology in Alzheimer´s diseases (AD). Besides the excessive NE release leading to the formation of amyloid plaques, the severity and duration of AD are also related to LC cell death, decreased cortical NE levels, and accelerated tau spread. An increase in NE release in the LC accompanied by reduced inflammatory markers was achieved in animal models using phasic invasive (i)VNS. In humans, phasic taVNS leads to an increase in pupil dilation, which can serve as a proxy for LC activation. Therefore, taVNS may have the potential to increase NE release and thereby impact LC function and related links to tau pathology.MethodTwenty‐four subjects (50% women; 22.96 +/‐ 2.24 years) received time‐synchronous phasic taVNS during a fixation task (∼ 18min) while recording pupillary dilation changes, which allowed systematic testing of phasic taVNS (3 sec) with low (3 mA) and high (5 mA) intensities and low (10 Hz) and high (25 Hz) frequencies at constant pulse width (250 µs).ResultDuring on stimulation pupil dilation was higher during real (0.18±0.03) as compared to sham (0.09±0.03) stimulation (χ2 = 10.29, p = 0.001) (see Fig. 1). Likewise higher (0.21±0.03) compared to lower (0.06±0.03) intensity (χ2 = 23.02, p < 0.001) led to higher pupil dilation, while there was no significant difference between higher (0.13±0.03) and lower (0.14±0.03) frequencies (χ2 = 0.11, p = 0.74).ConclusionSystematic testing of different frequencies and intensities, while keeping the pulse width constant, showed that phasic real taVNS and higher intensity lead to higher pupillary dilation, which is consistent with animal and human studies. Furthermore, the results suggest that the influence of intensity on pupil dilation might be stronger than that of frequency. The increased pupillary dilation during phasic taVNS could be an indication of LC involvement, however, more detailed studies e.g., functional magnetic resonance imaging (fMRI) during taVNS are needed to confirm this association.

Corydalis yanhusuo Polysaccharides Ameliorate Chronic Stress-Induced Depression in Mice through Gut Microbiota-Derived Short-Chain Fatty Acid Activation of 5-Hydroxytryptamine Signaling.

Depression, a prevalent psychiatric disorder, presents a serious health risk to humans. Increasing evidence suggested that the gut microbiota and the 5-hydroxytryptamine (5-HT) pathway both contribute significantly to depression. This research aimed to investigate how Corydalis yanhusuo polysaccharides (CYP) could potentially alleviate depression induced by chronic unpredictable mild stress in mice, as well as its underlying mechanism. The sucrose preference test, tail suspension test, and forced swimming test were employed to evaluate the behavior of mice. Enzyme-linked immunosorbent assay and PCR techniques were utilized to measure depression-related factors (dopamine [DA], 5-HT, norepinephrine [NE], brain-derived neurotrophic factor [BDNF], tryptophan hydroxylase 2 [TPH-2], 5-hydroxytryptophan [5-HTP], and tryptophan hydroxylase [TPH-1] levels). Hematoxylin and eosin staining and Nissl staining were conducted to observe histopathological changes in the hippocampus, the differences in the diversity of gut flora between groups were analyzed using 16S rRNA sequencing, and gas chromatography-mass spectrometry metabolomics was utilized to evaluate short-chain fatty acid (SCFA) concentrations. The findings indicated that CYP treatment increased the sucrose preference index, decreased the immobility time, and improved neuropathological injury. In depressed mice, CYP improved the dysregulation of the gut microbiota, and increased the SCFA levels. In addition, CYP enhanced the DA, 5-HT, NE, BDNF, and TPH-2 levels in the brain and the expression of 5-HTP and TPH-1 in the colon, while SCFAs were positively correlated with these levels. In summary, our study suggested that CYP may mitigate depression by ameliorating gut microbiota dysregulation, promoting the generation of SCFAs, and activation of 5-HT signaling expression.

Characteristics of Inflammatory Markers in Diabetic Foot Patients and Their Relationship With Prognosis of Diabetic Foot Ulcers

To explore the characteristics of baseline inflammatory markers in diabetic foot patients and their relationship with the prognosis of diabetic foot ulcers. The clinical data of diabetic foot patients (n=495) admitted to West China Hospital, Sichuan University since 2016 were retrospectively collected through the hospital electronic medical record system to analyze the characteristics of inflammatory markers and their relationship with the prognosis of diabetic foot ulcers. White blood cell count (WBC), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6) levels were significantly higher in patients defined as grade 4 on the Wagner Scale than those in patients defined as grade 0-3 on the Wagner Scale. Neutrophil percentage (NE%) was higher in Wagner grade-4 patients than those in Wagner grade-0 and grade-1 patients and higher in Wagner grade-3 patients than those in Wagner grade-0 patients. NE%, CRP, PCT, and IL-6 levels were positively correlated with the severity of diabetic foot, with the respective odds ratio (OR) at 95% confidence interval (CI) being 1.038 (1.019-1.056), 1.019 (1.012-1.026), 8.225 (2.015-33.576), and 1.017 (1.008-1.025). Using Wagner grade-0 patients as the reference, patients with higher WBC were more likely to progress to Wagner grade 2, 3, and 4, with the respective OR (95% CI) values being 1.260 (1.096-1.447), 1.188 (1.041-1.356), and 1.301 (1.137-1.490); patients with higher ESR were more likely to progress to Wagner grade 3 and 4, with the respective OR (95% CI) values being 1.030 (1.006-1.054) and 1.045 (1.019-1.071). Baseline ESR (P=0.008), CRP (P=0.039), and IL-6 (P=0.033) levels were lower in patients who had received antibiotics prior to their admission than those in patients who had not received antibiotics before admission. The levels of WBC, NE%, ESR, PCT, and IL-6 were lower in the full recovery group than those in the group of patients who did not respond to treatment. The higher the levels of NE% and IL-6, the worse the prognosis of diabetic foot ulcers became, with the respective OR (95% CI) values being 1.030 (1.010-1.051) and 1.008 (1.002-1.013). The severity of diabetic foot ulcers increased with the rise in baseline levels of inflammatory markers. Elevated baseline NE% and IL-6 levels suggest a poor prognosis. Our findings suggest that early assessment of diabetic foot infection and standardized antibiotic therapy should be implemented to improve the prognosis.

PCT, IL-6, and IL-10 facilitate early diagnosis and pathogen classifications in bloodstream infection

BackgroundIn the diagnosis of bloodstream infection (BSI), various inflammatory markers such as C-reactive protein (CRP), procalcitonin (PCT), interleukins (IL), white blood cell count (WBC), neutrophil percentage (NE%), platelet count (PLT), and erythrocyte sedimentation rate (ESR) have been extensively utilized. However, their specific roles in distinguishing BSI from local bacterial infection (LBI) and in classifying BSI pathogens remain uncertain.MethodsA historical cohort study was conducted, involving the enrollment of 505 patients with BSI and 102 patients with LBI. To validate the reliability of the clinical data obtained from this cohort, mouse models of BSI were utilized.ResultsOur findings revealed that patients with BSI had significantly higher levels of inflammatory markers, including CRP, PCT, IL-6, IL-10, WBC, NE%, and ESR, compared to those with LBI (p < 0.05). The receiver operating characteristic (ROC) curve analysis demonstrated that CRP, PCT, IL-6, IL-10, ESR and NE% exhibited excellent diagnostic efficacy for BSI. Additionally, we observed significant differences in CRP, PCT, IL-6, and IL-10 levels between patients with BSI caused by Gram-positive bacteria (GP-BSI) and Gram-negative bacteria (GN-BSI), but no significant variations were found among specific bacterial species. Furthermore, our study also found that CRP, PCT, and IL-10 have good discriminatory ability for vancomycin-resistant Enterococcus (VRE), but they show no significant diagnostic efficacy for other multidrug-resistant organisms (MDROs) such as carbapenem-resistant Enterobacteriaceae (CRE), carbapenem-resistant Pseudomonas aeruginosa (CRPA), and methicillin-resistant Staphylococcus aureus (MRSA). In our mouse model experiments, we observed a remarkable increase in PCT, IL-6, and IL-10 levels in mice with GN-BSI compared to those with GP-BSI.ConclusionOur study has confirmed that PCT, IL-6, and IL-10 are efficient biomarkers for distinguishing between BSI and LBI. Furthermore, they can be utilized to classify BSI pathogens and differentiate between VRE and vancomycin-susceptible Enterococcus. These findings are extremely valuable for clinicians as they enable timely initiation of empiric antibiotic therapies and ultimately lead to improved clinical outcomes for patients with BSI.

Abstract 17126: Role of Aldehydes, Transient Receptor Potential Ankyrin 1, and Catecholamines in Electronic Cigarette-Induced Endothelial Dysfunction in Mice

Although Electronic Nicotine Delivery Systems (ENDS)-derived aerosols induce endothelial dysfunction (ED), the mechanisms and constituents responsible for ED are unknown. Because ENDS use propylene glycol and vegetable glycerin (PG:VG) that thermally decompose into aldehydes, we hypothesized that acrolein and formaldehyde stimulate ED via the sensory transient receptor potential ankyrin-1 (TRPA1) receptor and the autonomic nervous system (ANS). To test this, female wild-type (WT) and TRPA1-null mice were exposed to either filtered air, PG:VG-derived aerosol, acrolein (AC; 1 ppm), or formaldehyde (FA, 5 ppm). Biomarkers of exposure (AC: 3HPMA; FA: 4TCA) and of harm (aortic ED; urinary catecholamines) were measured. The PG:VG and AC exposures significantly increased urinary 3HPMA levels; and the PG:VG and FA exposures significantly increased urinary 4TCA levels vs air controls. Both PG:VG and FA exposures (6h/d x 4d) in WT mice led to aortic ED (-64.6±3.8; –66.7±5.5 % ACh, respectively) vs air control (-81.5±1.5 % ACh); yet AC exposure did not induce ED (-76±2.9 % ACh). Exposures in TRPA1-null mice to PG:VG, AC, or FA did not lead to ED (-93.0±3.6; –82.8±3.1; -86.6±2.8 % ACh; respectively). As an index of ANS activation and a potential mediator of ED, urinary catecholamines (dopamine, DA; norepinephrine, NE; epinephrine, EPI; serotonin, 5HT) and metabolites were measured by UPLC-MS/MS in early (0-3h) and late (3-18h) post-exposure (PE) samples. In WT mice, PG:VG significantly increased early PE urine levels of NE (and two NE metabolites; +1.5-2.5x), DA metabolite (+1.5x), but not EPI nor 5HT; FA exposure increased DA (+3x), EPI (+2.5x), NE and NE metabolites (+1.2-2.5x), 5HT and 5HT metabolite (+2.5x) only in late PE; while AC exposure decreased early PE DA (-73%) and 5HT (-42%) levels. In TRPA1-null mice, PG:VG and AC exposures increased early PE levels of NE (+2x) and depressed EPI and 5HT levels (-40%); whereas FA exposure increased NE (+2x), EPI (+4x), and 5HT (+1.5) levels in late PE. Aldehydes induce ED through time-dependent mechanisms mediated in part by TRPA1 activation and ANS stimulation. These data indicate that ENDS use may increase cardiovascular disease risk via aldehydes generated by PG:VG independent of nicotine and flavorants.

Clinical Diagnostic Value of Serum GABA, NE, ET-1, and VEGF in Chronic Obstructive Pulmonary Disease with Pulmonary Hypertension.

Pulmonary hypertension (PH) is the one of the most common complications of chronic obstructive pulmonary disease (COPD). Whereas, the associated diagnostic factors are uncertain. The present study aims to investigate useful diagnostic factors in patients with COPD and PH (COPD-PH). A total of 111 patients with COPD in Shanxi Bethune Hospital from December 2019 to December 2020 were divided into COPD (PASP≤50 mmHg) and COPD-PH groups (PASP>50 mmHg). Pulmonary function and chest CT results were collected. Routine blood, biochemical, and blood coagulation function indices were examined for all patients. Arterial blood gas analysis and serum cytokines were also measured. Differences in the distribution of the above indicators between the two groups were analyzed using binary logistic regression analysis to identify the risk factors of COPD-PH, and multiple linear regression analysis to determine the factors affecting PASP. The influencing factors and joint predictive factors of the above linear regression analysis were analyzed using the ROC curve. The area under the curve and the best cut-off value were calculated, and their predictive value and clinical significance in disease diagnosis were discussed. A total of 27 indexes with statistically significant differences between the two groups were identified (P < 0.05). Binary Logistic regression analysis showed that the factors influencing the diagnosis of pulmonary hypertension were serum GABA, NE, VEGF, BUN, and LYM% levels (P < 0.05). Multiple linear regression showed that the factors influencing PASP were serum NE, ET-1, GABA, and VEGF levels, and the goodness of fit of the model was 0.748 (P < 0.001). ROC curve showed that the AUC of the combined diagnosis of serum NE, ET-1, GABA, and VEGF levels was 0.966 (sensitivity, 87.5%; specificity, 93.65%). Serum NE and ET-1 are risk factors for COPD-PH, whereas serum GABA and VEGF are protective factors against COPD-PH. The combined diagnostic value of serum NE, ET-1, GABA, and VEGF levels was the highest.

Assessment of the risk for cholecystitis when performing laparoscopic cholecystectomy in a retrospective cohort study

Background/Aim: Acute cholecystitis most often occurs as an acute exacerbation of chronic cholecystitis. In over 90% of patients, the primary factor in the development of acute cholecystitis is the obstruction of the gallbladder or cystic duct with an impacted calculus. In less than 10% of patients, acute cholecystitis can be attributed to other causes: direct trauma to the biliary tract, torsion of the gallbladder, twisting of the vascular loop after surgical procedures on the abdominal organs, and Salmonella typhi infection. The aim of this study is to compare preoperative variables in patients with acute cholecystitis, which are not only important for diagnosis but also have significance in determining the severity of acute cholecystitis in the preoperative period. Methods: This retrospective systematic research included all clinical cases diagnosed with acute cholecystitis from January 2019 to December 2019, totaling 56 patients at CHC Kosovska Mitrovica. For the study, data from medical records were used for statistical analysis. Results: A total of 56 patients (mean age 53 years; 26 men and 30 women) were included in this study. Among them, 32 patients (57.15%) had characteristics of simple cholecystitis, while 24 patients (42.85%) had severe cholecystitis. The group with severe cholecystitis consisted of older patients, with equal representation of both sexes, and higher levels of WBC, NE%, PLT, ALT, GGT, total bilirubin, and CRP (P&lt;0.05). Imaging studies showed that the group with severe cholecystitis exhibited significantly more wall distension, particularly in the stratified wall, compared to the group with simple cholecystitis (P&lt;0.05). Severe cholecystitis was associated with statin use (79.2%) and triglyceride values of 1.55 (0.47); both variables showed a statistically significant association with severe cholecystitis (P&lt;0.05). Conclusion: It is extremely important to skillfully identify patients with simple or severe forms of acute cholecystitis. Possible solutions include organizing campaigns to raise public awareness for faster consultations in cases of acute abdominal pain, establishing universal health coverage (diet), and improving technical platforms.