CTRL Group Research Articles

The Effect of Rubus idaeus Polyphenols Extract in Induced Endometriosis in Rats.

Endometriosis is a common gynecological condition with a complex physio-pathological background. This study aimed to assess the role of Rubus idaeus leaf extract (RiDE) as a potential therapeutic agent in reducing the size of the endometriotic lesions and modulate the plasma expression of MMP-2, MMP-9, and TGF-β1. The endometriotic lesions were induced in a rat model by the autologous transplant of endometrium. Thirty-six female rats, Wistar breed, with induced endometriosis, were divided into four groups and underwent treatment for 28 days. The CTRL group received 0.5 mL/day of the vehicle; the DG group received 1 mg/kg b.w./day dienogest; the RiDG group received 0.25 mL/kg b.w./day RiDE and the D+RiDG group received 1 mg/kg b.w./day dienogest and 0.25 mL/kg b.w./day RiDE, respectively. Rats' weight, endometriotic lesion diameter and grade, and plasma levels of MMP-2, MMP-9, and TGF-β1 were assessed before and after treatment. The administration of RiDE in association with dienogest vs. dienogest determined a lower weight gain and a reduction in diameter of the endometriotic lesions. RiDE administration restored MMP2 and MMP9 plasma levels to initial conditions. Rubus idaeus extract may help in reducing dienogest-associated weight gain, lower the size of endometriotic lesions, and have anti-inflammatory effects through MMP2 and MMP9 reduction.

Association of SGLT2 inhibitor dapagliflozin with risks of acute kidney injury and all-cause mortality in acute myocardial infarction patients

ObjectiveSodium-glucose cotransporter 2 (SGLT2) inhibitors have well-documented effects in reducing hospitalization or cardiovascular mortality, while the association of SGLT2 inhibitor dapagliflozin (DAPA) and the risk of acute kidney injury (AKI) in acute myocardial infarction (AMI) patients has not been comprehensively investigated. Therefore, we aimed to assess the association between DAPA and AKI risk in AMI patients after percutaneous coronary intervention (PCI) therapy.MethodsUsing the Changzhou Acute Myocardial Infarction Registry database, we retrospectively included AMI patients from January 2017 to August 2021 and analyzed the risk of AKI and all-cause mortality after PCI therapy. The patients were divided into two groups according to the use of DAPA (DAPA group and Ctrl group). Patients in the DAPA group started to use DAPA after admission and continued its use during hospitalization and follow-up period. Baseline characteristics were balanced between the two groups with a propensity score matching (PSM) analysis. The outcome was AKI within 7 days after PCI and all-cause mortality during a follow-up of 2 years. Univariate and multivariate logistic regression analyses were used to assess the association between DAPA and AKI risk.ResultsA total of 1839 AMI patients undergoing PCI were enrolled. DAPA was used in 278 (15.1%) patients. Postoperative AKI occurred in 351 (19.1%) cases. A 1:1 PSM analysis was used to reduce confounding factors. The multivariate stepwise regression analysis showed that DAPA (odds ratio, OR 0.66; 95% confidence interval, CI 0.44–0.97; P = 0.036) was an independent protective factor in the entire cohort. After matching, the use of DAPA in AMI patients was independently associated with a decline of AKI risk (OR 0.32; 95% CI, 0.19–0.53; P < 0.001) after hospital admission. Meanwhile, there were significant differences in mortality between the DAPA group and Ctrl group (2.5% vs. 7.6%, P = 0.012).ConclusionSGLT2 inhibitor DAPA was associated with lower risks of incident AKI and all-cause mortality in AMI patients after PCI therapy.

Improvement of Mouse Preantral Follicle Survival and Development following Co-Culture with Ovarian Parenchyma Cell Suspension.

The parallel and continued improvements in both infertility treatment and the management of malignancy cases have brought to the forefront the potential for fertility preservation. Using ovarian follicular resources can effectively improve reproductive capacity and prevent infertility. The primary aim of this research was to try to generate an appropriate in vivo environment for the growth of the mouse follicles. Hence, the possible effects of the ovarian parenchyma cell suspension were explored on the growth and maturation of preantral follicles in vitro. In this experimental study, ovarian parenchymal cells were mechanically dissociated from preantral follicles of 12-14 days-old NMRI mice and then divided into 5 experimental groups (G1: Control, G2: Fresh follicle with fresh parenchyma cell suspension, G3: Vitrified-warmed follicle with fresh parenchyma cell suspension, G4: Fresh follicle with frozen-thawed parenchyma cell suspension, and G5: Vitrified-warmed follicle with frozenthawed parenchyma cell suspension). The diameter of the follicles and immature oocytes, viability, antrum formation, resumption of meiosis, in vitro fertilization (IVF), and Gdf9, Bmp6, and Bmp15 gene expression were examined on different periods. The diameter of the follicles and the oocytes on days 4 and 8, as well as the survival rate of the follicles up to day 12, were significantly higher in G2 and G4 compared to the Ctrl group (G1: 73.66%, G2:87.99%, G3: 82.70%, G4: 94.37%, and G5: 78.59%). Expression of growth marker genes for G3, and G5 groups was significantly higher than other groups, which indicated the protective effects of parenchyma cell suspension on follicles damaged by vitrification solutions. The growth, survival, and maturation of preantral follicles could be enhanced by co-culturing them with ovarian parenchyma cells. Further studies are needed to optimize the conditions for a successful parenchyma cell suspension-induced in vitro maturation (IVM) to occur in infertility clinics.

Effect of pretreatment with Devil's Claw on locomotor activity, infarct volume, and neuronal density in focal cerebral ischemia in rats.

Stroke is a highly prevalent and devastating condition affecting millions worldwide. The Devil's Claw (DCW) plant is a native African plant whose anti-inflammatory, antioxidant, and neuroprotective properties have been investigated. We postulated that DCW could protect the brain injury caused by cerebral ischemia. The rats were randomly divided into four groups. The sham and control (Ctrl) groups received pretreatment with a distilled water vehicle. Doses of 200 and 400 mg/kg were selected for pretreatment with DCW. The filament or intravascular occlusion method was used for middle cerebral artery occlusion (MCAO). The Triphenyl tetrazolium chloride (TTC) staining method was used to investigate the infarct zone and penumbra volume. The neuroprotective effect of DCW was measured by hematoxylin staining. Movement performance was evaluated from neurological deficit score, rotarod performance, and open field tests. TTC staining showed that the DCW/400 group could maintain the penumbra's structure and reduce the infarct volume compared to the Ctrl group (p<0.001). Histological studies confirmed the neuroprotective properties of DCW at doses of 200 and 400 mg/kg compared to the Ctrl group (p<0.01 and p<0.0001, respectively). The results of behavioral tests showed an improvement in behavioral performance in pretreatment 400 mg/kg doses compare to Ctrl group (p<0.0001). The study showed that pretreatment with DCW with its neuron protection potential reduces the infarct area and restores motor function after MCAO.

Cryptosporidium parvum infection alters the intestinal mucosa transcriptome in neonatal calves: implications for immune function.

One of the leading causes of infectious diarrhea in newborn calves is the apicomplexan protozoan Cryptosporidium parvum (C. parvum). However, little is known about its immunopathogenesis. Using next generation sequencing, this study investigated the immune transcriptional response to C. parvum infection in neonatal calves. Neonatal male Holstein-Friesian calves were either orally infected (N = 5) or not (CTRL group, N = 5) with C. parvum oocysts (gp60 subtype IIaA15G2R1) at day 1 of life and slaughtered on day 7 after infection. Total RNA was extracted from the jejunal mucosa for short read. Differentially expressed genes (DEGs) between infected and CTRL groups were assessed using DESeq2 at a false discovery rate < 0.05. Infection did not affect plasma immunohematological parameters, including neutrophil, lymphocyte, monocyte, leucocyte, thrombocyte, and erythrocyte counts as well as hematocrit and hemoglobin concentration on day 7 post infection. The immune-related DEGs were selected according to the UniProt immune system process database and were used for gene ontology (GO) and pathway enrichment analysis using Cytoscape (v3.9.1). Based on GO analysis, DEGs annotated to mucosal immunity, recognizing and presenting antigens, chemotaxis of neutrophils, eosinophils, natural killer cells, B and T cells mediated by signaling pathways including toll like receptors, interleukins, tumor necrosis factor, T cell receptor, and NF-KB were upregulated, while markers of macrophages chemotaxis and cytosolic pattern recognition were downregulated. This study provides a holistic snapshot of immune-related pathways induced by C. parvum in calves, including novel and detailed feedback and feedforward regulatory mechanisms establishing the crosstalk between innate and adaptive immune response in neonate calves, which could be utilized further to develop new therapeutic strategies.

Transcriptomics-based analysis reveals the nephrotoxic effects of triphenyltin (TPT) on SD rats by affecting RAS, AQPs and lipid metabolism

Triphenyltin (TPT) is a class of organotin compounds that are extensively used in industry and agriculture. They have endocrine-disrupting effects and cause severe environmental contamination. Pollutants may accumulate in the kidneys and cause pathological complications. However, the mechanism of TPT's toxicological effects on the kidney remains unclear. This study aimed to investigate the toxic effects and mechanism of action of TPT exposure on renal impairment in rats. Male SD rats were divided into four groups: the Ctrl group (control group), TPT-L group (0.5 mg/kg/d), TPT-M group (1 mg/kg/d), and TPT-H group (2 mg/kg/d). After 28 days of exposure to TPT, we observed the morphology and structure of kidney tissue using HE, PASM, and Masson staining. We also detected serum biochemical indexes, performed transcriptome sequencing of rat kidney tissue using RNA-seq. Furthermore, protein expression levels were measured through immunohistochemistry and gene expression levels were determined using RT-qPCR. The study results indicated a decrease in kidney weight and relative kidney weight after 28 days of exposure to TPT. Additionally, TPT caused damage to kidney structure and function, as evidenced by HE staining, PASM staining, and serum biochemical tests. Transcriptomics identified 352 DEGs, and enrichment analyses revealed that TPT exposure primarily impacted the renin-angiotensin system (RAS). The expression levels of water channel proteins were reduced, and the expression levels of RAS and lipid metabolism-related genes (Mme, Ace, Fasn, Cyp4a8, Cpt1b and Ppard) were significantly decreased in the TPT-treated group. In summary, exposure to TPT may impair renal structure and function in rats by affecting RAS, AQPs, and lipid metabolism.

Pull-Up Performance Is Affected Differently by the Muscle Contraction Regimens Practiced during Training among Climbers.

Sport climbing performance is highly related to upper limb strength and endurance. Although finger-specific methods are widely analyzed in the literature, no study has yet quantified the effects of arm-specific training. This study aims to compare the effects of three types of training involving different muscle contraction regimens on climbers' pull-up capabilities. Thirty advanced to high-elite climbers were randomly divided into four groups: eccentric (ECC; n = 8), isometric (ISO; n = 7), plyometric (PLYO; n = 6), and no specific training (CTRL; n = 9), and they participated in a 5-week training, twice a week, focusing on pull-ups on hangboard. Pre- and post-training assessments were conducted using a force-sensing hangboard, analyzing force, velocity, power, and muscle work during three pull-up exercises: pull-ups at body weight under different conditions, incremental weighted pull-ups, and an exhaustion test. The CTRL group showed no change. Maximum strength improved in all three training groups (from +2.2 ± 3.6% to +5.0 ± 2.4%; p < 0.001); velocity variables enhanced in the ECC and PLYO groups (from +5.7 ± 7.4 to +28.7 ± 42%; p < 0.05), resulting in greater power; amplitude increased in the ECC group; and muscle work increased in the PLYO group (+21.9 ± 16.6%; p = 0.015). A 5-week training period effectively enhanced arm performance, but outcomes were influenced by the chosen muscle contraction regimens and initial individual characteristics.

Paris saponin VII promotes ferroptosis to inhibit breast cancer via Nrf2/GPX4 axis

Breast cancer (BC) is one of the malignancies threatening the woman's health. Our study aims to explore the underlying mechanism behind the anti-tumor function of Paris saponin VII (PS VII) in BC. Xenografting experiment was conducted to monitor the tumor growth. The Ki67 and 4-HNE expression were analyzed via immunohistochemical assay. After different treatments, the cell viability, proliferation, invasion, and migration capacity of BC cells were measured by the CCK-8, colony formation, transwell, and wound healing assays, respectively. The ratio of GSH/GSSG was measured by the GSH/GSSG ratio detection assay kit. The lipid ROS and Fe2+ levels were quantified by flow cytometry analysis. The expressions of TFR1, ACSL4, Nrf2, and GPX4 were measured via western blotting. Compared with the Ctrl group, the tumor volumes, and Ki67 expression were markedly reduced in PS VII groups, and the BC cell viability was decreased by PS VII treatment in a dose-dependent manner. The colony numbers, invasive cells, and migration rates were also significantly decreased by PS VII treatment. Then, the Nrf2 as well as GPX4 expressions were decreased and TFR1 expression was increased by PS VII treatment in vitro and in vivo, while there was no difference in ACSL4 expression between Ctrl and PS VII groups. Moreover, the above effects of PS VII could not be observed in GPX4 knockdown cells. PS VII can promote ferroptosis to inhibit BC via the Nrf2/GPX4 axis, which innovatively suggests the pro-ferroptosis effect and therapeutic potential of PS VII in BC.

Impact of try-in paste removal on the fatigue behavior of bonded lithium disilicate ceramics

This in vitro study assessed the effectiveness of three cleaning protocols (air-water spray, 37% phosphoric acid, or Ivoclean) on lithium disilicate restorations' fatigue behavior after try-in paste application, compared to a clean condition. Lithium disilicate discs (IPS e.max CAD, Ivoclar) with Ø-= 12 mm and 1 mm thickness were prepared from prefabricated CAD-CAM blocks, polished, subjected to CAD-CAM milling topography simulation and crystallization. After, etching with 5% hydrofluoric acid and the application of try-in paste (Variolink try-in paste shade white; load of 2.5 N for 5 min) was performed. Discs that received try-in paste were divided into three groups according to the removal protocol: SPRAY – air-water spray for 30 s; HPO – active application of 37% phosphoric acid for 60 s; IVOC – application of Ivoclean for 20 s. Control group (CTRL group) did not receive the try-in paste application. Half of the specimens (n= 15) were tested in the baseline condition (24 h up to 7 days), and the others underwent 25,000 thermal cycles (5 – 55 °C) + 210 days of distilled water storage (37 °C). Additional specimens (n= 3) underwent monotonic testing (1 mm/min). Fatigue testing involved a cyclic fatigue approach (20 Hz, initial load = 100 N – 5000 cycles, step size = 50 N – 10,000 cycles) until a visible crack appeared. Fractographic and topographic analyses were performed. Fatigue data were statistically analyzed with two-way ANOVA, Kaplan-Meier log-rank (Mantel-Cox), and independent t-test (α= 0.05). In the baseline condition, the IVOC group resulted in a superior fatigue behavior compared to the CTRL and SPRAY groups, but similar to the HPO group. The HPO and SPRAY presented a similar fatigue behavior to the CTRL group. It was noticed a decrease in fatigue behavior after aging, which resulted in all the cleaning protocols leading to similar fatigue behavior compared to the CTRL group. On the SPRAY group surface, try-in pastes remnants were noticed. In summary, despite a detrimental impact at baseline conditions, all tested cleaning protocols seem proper to remove the try-in paste from the ceramic's surface in the long-term evaluation.

Vacuum sealing drainage technique combined with carbon nanomaterial dressings applied in the treatment of skin wounds.

Limb trauma (LT) encompasses a variety of conditions and currently poses certain therapeutic challenges in clinical practice. To assess the effectiveness of applying the vacuum sealing drainage (VSD) technique combined with carbon nanomaterial (CNM) dressings in the nursing treatment of LT patients. Eighty-six LT patients were enrolled into a control (Ctrl) group and an experimental (Exp) group. Patients in the Ctrl group underwent conventional wound cleansing and drainage methods with regular dressings, while those in the Exp group received VSD combined with CNM dressings. Both groups received the same nursing procedures. Various indicators were observed and compared to analyse the final outcomes. The Exp group exhibited a higher favourable wound healing (FWH) rate, overall limb functional recovery (LFR) rate, and nursing satisfaction compared to the Ctrl group (p < 0.05). Meanwhile, patients in the Exp group experienced a greatly lower incidence of complications (IOC), pain scores, and average hospital stays (HS), presenting statistically/significant differences compared to those in the Ctrl group (p < 0.05). The combination of the VSD technique with CNM dressings demonstrated advantages in nursing treatment of LT patients. This approach could enhance rates of FWH, reduce IOC, alleviate postoperative pain, facilitate LFR, and shorten HS. Furthermore, patients in the Exp group expressed higher nursing satisfaction. Results of this work yielded a novel therapeutic option for clinical practice, enhancing treatment outcomes and rehabilitation processes for LT patients.

Drug sensitivity testing during dental pulp-dentin complex repair: its effect on treatment outcome and prognosis.

To assess the efficacy of targeted therapy based on drug sensitivity testing (DST) results in patients with acute pulpitis (AP). A total of 80 cases of AP were included retrospectively and divided into two groups: control (Ctrl) group (conventional drug palliative therapy, n=40) and experimental (Exp) group (DST + non-resistant drugs, n=40). The clinical data and laboratory examination data of patients, including bacterial culture data, drug sensitivity test results, Ca and P contents in dental pulp, visual analogue scale (AVS), treatment satisfaction, and dental pulp incidence, were collected and analyzed. Prevotella melaninogenica, Fusobacterium nucleatum, and Porphyromonas gingivalis exhibited higher resistance rates (RS) to penicillin and amoxicillin but no resistance to imipenem and metronidazole. The content of Ca and P in the dental pulp of the Exp group patients was significantly higher than that of the Ctrl group (P=0.006). The total response rate (95% vs. 77.5%, P=0.018) and overall patient satisfaction (92.5% vs. 80%, P=0.021) were also significantly higher in the Exp group than in the Ctrl group. Additionally, when the follow-up duration exceeded 1 year, the incidence of dental pulp reactions in the Exp group was significantly lower than that of the Ctrl group (P=0.026). These findings suggest that Gram-negative anaerobes are the predominant oral pathogens in patients with AP. Imipenem and metronidazole demonstrate the most effective anti-infective properties against these anaerobes. Utilizing DST to select non-resistant drugs for treatment prior to therapy effectively enhances clinical outcomes for patients with AP.

Clinical evaluation of continuous irrigation with Bikerui combined with negative pressure closed drainage and platelet-rich plasma technique for treatment of stage III-IV pressure ulcer.

As a catastrophic complication of bedridden and elderly patients, pressure ulcer usually continuously affects patients' health and quality of life, so the daily care of wounds is attached great importance in clinic. This work investigated the effect of Bikerui disinfectant + vacuum sealing drainage (VSD) + platelet-rich plasma (PRP) therapy on patients with stage III ~ IV pressure sore. In this work, 110 patients with pressure ulcer (PU) treated in our hospital were enrolled and were randomly divided into an experimental group (Exp group) and a control group (Ctrl group) by a blind selection method, with 55 cases in each group. Patients in the Ctrl group received surgical debridement + VSD for treatment, while those in the Exp group were treated with Bikerui disinfectant + VSD + PRP. Inflammatory response (IR) score, PU healing (PUH) score, healing time, dressing change frequency (DCF), dressing interval time (DIT), and scar recovery (SR) score of patients in different groups were compared. The results revealed that the positive rate of bacterial culture in wound secretions in the Exp group was greatly lower than that in the Ctrl group 1 or 2 weeks after treatment (p < 0.05). The IR score in the Exp group was much lower at week 1 and 2 after treatment (p < 0.05). The total effective rate (TER) in the Exp group was obviously higher than that in the Ctrl group (94.55% vs. 76.36%). The results suggested that Bikerui disinfectant + VSD + PRP therapy could effectively improve the inflammatory degree of PU patients, promote the wound repair and scar recovery of patients, and greatly improve the clinical efficacy of PU patients.

Levels of serum inflammatory cytokines and their correlations with disease severity in patients with chronic spontaneous urticaria.

Inflammation is crucial in the pathogenesis of chronic spontaneous urticaria (CSU). Investigating the correlation between levels of serum inflammatory cytokines (SICs) and the severity of CSU is of great significance for understanding the disease mechanism and finding effective treatment strategies. In this context, this work was developed. This work involved a researchy group (Res group) of 114 patients with CSU and a control group (Ctrl group) of 100 healthy individuals. SICs including leukotriene B4 (LTB4), leukotriene C4 (LTC4), interleukin (IL) 4 (IL-4), IL-17, IL-31, and tumor necrosis factor-γ (TNF-γ), of patients in different groups were measured and compared. Furthermore, the correlations between each SIC and pruritus severity, duration of pruritus, urticaria activity, and quality of life (QOL) were compared among the patients in different groups. The Res group exhibited higher levels of LTB4, LTC4, IL-4, IL-17, and IL-31 but lower levels of TNF-γ. Great differences (p < 0.05) were found in IL-4, IL-17, and IL-31 among the patients with different pruritus severity, and positive correlations were observed between IL-17 and IL-31 levels and urticaria activity in the patients (p < 0.05). Additionally, levels of IL-4 and IL-31 exhibited a positive association to QOL scores in the patients, with obvious differences (p < 0.05). IL-4, IL-17, and IL-31 showed the strongest correlation with the severity of CSU, which may be attributed to their involvement in immune, inflammatory, and pruritic reactions, exacerbating the disease condition.

Retinal neurotoxicity induced by inhalation of isopropyl nitrite

Purpose: Popper is a slang term given broadly to drugs of the chemical class called alkyl nitrites that are inhaled. Most widely sold products include the original isopropyl nitrite (IPN) a mix of isopropyl alcohol with nitrous acid and sulphuric acid. Multiple cases of retinopathy have been reported since 2001 that have been associated with the inhalation of poppers, predominantly in Europe. However, no clear information about the effect of these inhalant agents on retinal cells have been shown to date. Our work tries to address the neurotoxic effect of IPN on outer retinal cells evidenced by immunocytochemical technique on murine model.Methods: 8 Opn4‐KO mice were used, n:4 were treated with IPN using a aero chamber inhalator for 20 days (1 mL/5 s) and n:4 with oxygen (ctrl group). CARR antibody was used, encoded by the Arr3 gene in murine species. All the animals were supplied by the Animal Experimentation Center of the UAH. The euthanasia of the animals was carried out in accordance with RD 53/2013 Spanish regulations.Results: The sagittal sections of the Opn4 mouse retina taken with confocal microscopy at 40×, the expression of arrestin‐C can be observed, predominantly in the internal and external segments of the cones without disruption, the presence of CARR confirms the presence of cells of retinal cone in our CARR cuts (Cy5, red), The exposure parameters of the samples and image capture in the confocal microscope were taken based on the immunoreactivity in the Opn4 animal model, remaining stable for the rest of the photos. Labelling at the level of the outer plexiform layer is equally intense in Opn4 Ctrl and Opn4 to animals.Conclusions: The results failed to demonstrate morphological changes of clinical importance that could determine a neurotoxic role in the retina of IPN inhalation; however, we propose the existence of other factors and/or pharmacological interactions with the use of poppers in the aetiopathogenesis of this new ophthalmological disease.

Berberine attenuates TNBS-induced colitis in mice by improving the intestinal microbiota.

To investigate the effects of berberine (BBR) as a treatment on intestinal microecological alterations and enteritis in mice produced by TNBS. There were seven mice per group: seven in the healthy group (Ctrl), seven in the TNBS-induced enteritis group (TNBS), and seven in the berberine treatment group (BBR). The mice were weighed, slaughtered after 7 days, and subjected to high-throughput intestinal microecological analysis by Illumina, as well as haematological detection and imaging evaluation of colon pathology. The alterations in colon length, immune cell subpopulations, inflammatory factors, and intestinal microecology of mice induced by BBR were refined using a battery of experiments and observations. According to intestinal microecological studies, BBR can increase the number of bacteria, including Lactobacillus, Verrucomicrobia, Bacteroides, and Akkermansia muciniphila. BBR has a therapeutic effect on TNBS-induced colitis in mice, which is associated with modifications in immune cell subpopulations and intestinal microecology. It also offers a viable approach as a prospective probiotic (like Akkermansia muciniphila) to IBD therapy in clinical settings.

Increased NAD+ levels protect female mouse reproductive system against zearalenone-impaired glycolysis, lipid metabolism, antioxidant capacity and inflammation

The reproductive system is a primary target organ for zearalenone (ZEN, a widespread fusarium mycotoxin) to exert its toxic effects, including decreased antioxidant capacity and aggravated inflammatory response. These ZEN-induced reproductive abnormalities are partially caused by the declining levels of nicotinamide adenine dinucleotide (NAD+), which results in an imbalance in lipid/glucose metabolism. Accordingly, the present study aimed to investigate whether supplements of nicotinamide mononucleotide (NMN, a NAD+ precursor) in female mice could protect against ZEN-induced reproductive toxicity. In this study, thirty female mice were randomly divided into three groups that were intragastrically administered with i) 0.5% DMSO (the Ctrl group), ii) 3 mg/(kg bw.d) ZEN (the ZEN group), or iii) ZEN + 500 mg/(kg bw.d) NMN (the ZEN/NMN group) for two weeks. The results revealed that, compared with the Ctrl group, animals exposed to ZEN exhibited reproductive toxicity, such as decreased antioxidant capacity and aggravated inflammatory response in reproductive tissues. These effects were strongly correlated with lower activities in key glycolytic enzymes (e.g., ALDOA and PGK), but increased expressions in key lipid-synthesis genes (e.g., LPIN1 and ATGL). These changes contribute to lipid accumulation, specifically for diacylglycerols (DAGs). Furthermore, these ZEN-induced changes were linked with disturbed NAD+ synthesis/degradation, and subsequently decreased NAD+ levels. Notably, NMN supplements in mice protected against these ZEN-induced reproductive abnormalities by boosting NAD+ levels. Herein, the present findings demonstrate that potential strategies to enhance NAD+ levels can protect against ZEN-induced reproductive toxicity.

Pyrroloquinoline quinone supplementation attenuates inflammatory liver injury by STAT3/TGF-β1 pathway in weaned piglets challenged with lipopolysaccharide.

This study is aimed to evaluate the effect and underling mechanism of dietary supplementation with pyrroloquinoline quinone (PQQ) disodium on improving inflammatory liver injury in piglets challenged with lipopolysaccharide (LPS). A total of seventy-two crossbred barrows were allotted into four groups as follows: the CTRL group (basal diet + saline injection); the PQQ group (3 mg/kg PQQ diet + saline injection); the CTRL + LPS group (basal diet + LPS injection) and the PQQ + LPS group (3 mg/kg PQQ diet + LPS injection). On days 7, 11 and 14, piglets were challenged with LPS or saline. Blood was sampled at 4 h after the last LPS injection (day 14), and then the piglets were slaughtered and liver tissue was harvested. The results showed that the hepatic morphology was improved in the PQQ + LPS group compared with the CTRL + LPS group. PQQ supplementation decreased the level of serum inflammatory factors, aspartate aminotransferase and alanine transaminase, and increased the HDL-cholesterol concentration in piglets challenged with LPS; piglets in the PQQ + LPS group had lower liver mRNA level of inflammatory factors and protein level of α-smooth muscle actin than in the CTRL + LPS group. Besides, mRNA expression of STAT3/TGF-β1 pathway and protein level of p-STAT3(Tyr 705) were decreased, and mRNA level of PPARα and protein expression of p-AMPK in liver were increased in the PQQ + LPS group compared with the CTRL + LPS group (P < 0·05). In conclusion, dietary supplementation with PQQ alleviated inflammatory liver injury might partly via inhibition of the STAT3/TGF-β1 pathway in piglets challenged with LPS.

Effects and mechanism of anlotinib on radiosensitivity of non-small cell lung cancer cell lines a549

To investigate the effect and mechanism of Anlotinib on radio sensitization of human lung adenocarcinoma cell line A549. Human lung adeno carcinoma cell line A549 was teated with anlotinib and/or radiotherapy, then divided four groups, control group (Ctrl), Anlotinib treatment group (A), irradiation group (RT) and Anlotinib combined with irradiation group (A+RT) . CCK8 method was used to determine cell proliferation; the clone formation experiment was used to determine the inhibitory effect on cell growth; flow cytometry was used to determine cell cycle and apoptosis; immunofluorescence of γ-H2AX was used to determine DNA damage; expression of DNA-PKcs were detected by Western blot. Anlotinib inhibited proliferation and clonogenic survival following irradiation. The dose (Dq), the average lethal dose (D0) and the survival score (SF2) in the anlotinib combined radiotherapy group was significantly lower than those in the radiotherapy group. Anlotinib decreased G2/M phase arrest and promoted the cells apoptosis induced by in irradiation. The confocal microscopy results showed the average number of γ-H2AX foci in the A+RT group was more than that in RT group. The protein levels of DNA-PKcs were higher in A+RT group than that in RT group. Anlotinib enhances the radio sensitivity of A549 cells, which may be attributed to the delay DNA damage repair. It provides a rationale strategy by Anlotinib combined with irradiation for NSCLC.

Combined RhoA morpholino and ChABC treatment protects identified lamprey neurons from retrograde apoptosis after spinal cord injury.

Previously, we reported that RhoA knockdown by morpholino antisense oligonucleotides (MOs), and enzymatic digestion of chondroitin sulfate proteoglycans (CSPGs) at the site of injury with chondroitinase ABC (ChABC), each can reduce retrograde neuronal apoptosis after spinal cord transection in the lamprey. To elucidate the mechanisms in neuronal survival and axon regeneration, we have investigated whether these two effects are additive in vivo. We used lampreys as a spinal cord injury model. MOs were used to knockdown RhoA and Chondroitinase ABC (ChABC) was used to digest CSPGs in vivo. Retrograde labeling, fluorochrome-labeled inhibitor of caspase activity (FLICA), immunohistochemistry, and western blots were performed to assess axonal regeneration, neuronal apoptotic signaling and Akt activation. Four treatment combinations were evaluated at 2-, 4-, and 10-weeks post-transection: (1) Control MO plus enzyme buffer (Ctrl); (2) control MO plus ChABC; (3) RhoA MO plus enzyme buffer (RhoA MO); and (4) RhoA MO plus ChABC (RhoA MO + ChABC). Consistent with our previous findings, at 4-weeks post-transection, there was less caspase activation in the ChABC and RhoA MO groups than in the Ctrl group. Moreover, the RhoA MO plus ChABC group had the best protective effect on identified reticulospinal (RS) neurons among the four treatment combinations. At 2 weeks post-transection, when axons have retracted maximally in the rostral stump and are beginning to regenerate back toward the lesion, the axon tips in the three treatment groups each were closer to the transection than those in the Ctr MO plus enzyme buffer group. Long-term axon regeneration also was evaluated for the large, individually identified RS neurons at 10 weeks post-transection by retrograde labeling. The percent regenerated axons in the RhoA MO plus ChABC group was greater than that in any of the other groups. Akt phosphorylation levels at threonine 308 was quantified in the identified RS neurons by western blots and immunofluorescence. The RhoA MO plus ChABC treatment enhanced pAkt-308 phosphorylation more than any of the other treatment groups. Although some of the effects of CSPGs are mediated through RhoA activation, some growth-inhibiting mechanisms of RhoA and CSPGs are independent of each other, so combinatorial therapies may be warranted.

Peripheral artery disease causes consistent gait irregularities regardless of the location of leg claudication pain

BackgroundThe most common symptom of peripheral artery disease (PAD) is intermittent claudication that involves the calf, thigh, and/or buttock muscles. How the specific location of this leg pain is related to altered gait, however, is unknown. ObjectivesWe hypothesized that because the location of claudication symptoms uniquely affects different leg muscle groups in people with PAD, this would produce distinctive walking patterns. MethodsA total of 105 participants with PAD and 35 age-matched older volunteers without PAD (CTRL) were recruited. Participants completed walking impairment questionnaires (WIQ), Gardner-Skinner progressive treadmill tests, the six-minute walk test, and we performed an advanced evaluation of the biomechanics of their overground walking. Participants with PAD were categorized into 4 groups according to their stated pain location(s): calf only (C, n = 43); thigh and calf (TC, n = 18); buttock and calf (BC, n = 15); or buttock, thigh, and calf (BTC, n = 29). Outcomes were compared between CTRL, C, TC, BC and BTC groups using a one-way ANOVA with post-hoc comparisons to identify and assess statistically significant differences. ResultsThere were no significant differences between CTRL, C, TC, BC and BTC groups in distances walked or walking speed when either pain-free or experiencing claudication pain. Each participant with PAD had significantly dysfunctional biomechanical gait parameters, even when pain-free, when compared to CTRL (pain-free) walking data. During pain-free walking, out of the 18 gait parameters evaluated, we only identified significant differences in hip power generation during push-off (in C and TC groups) and in knee power absorption during weight acceptance (in TC and BC groups). There were no between-group differences in gait parameters while people with PAD were walking with claudication pain. ConclusionsOur data demonstrate that PAD affects the ischemic lower extremities in a diffuse manner irrespective of the location of claudication symptoms. Database RegistrationClinicalTrials.gov NCT01970332.