Inhibitory effects of total saponins from Ilex pubescens Hook against hydrogen peroxide-induced cardiomyocyte apoptosis

Purpose: To study the protective effects of total saponins from Ilex pubescens Hook (IPTS) against cardiomyocyte apoptosis.Methods: Response surface methodology (RSM) based on Box-Benhnken Design (BBD) was carried out to optimize the extraction of IPTS. Thereafter, H9c2 cell model prepared by hydrogen peroxide (H2O2) treatment was used to investigate the effects of IPTS on cardiomyocyte apoptosis. Cell viability was determined using MTT assay, while the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), creatine kinase (CK) and catalase (CAT) were measured as indices of oxidative stress. Expressions of proteins related to apoptosis (caspase-3, Bax and Bcl-2) were measured using Western blot assay.Results: Optimal IPTS extraction was achieved with extraction temperature of 86.6 °C, extraction time of 2.23 h and water: raw material ratio of 10.8 mL/g. IPTS extract, at doses of 200, 400, 600 and 800 μg/mL, significantly increased the viability of H2O2-treated H9c2 cells (p < 0.05), but significantly decreased LDH and CK activities (p < 0.01). It also led to significant increases in SOD and CAT activities, and significant decreases in the levels of MDA in these cells (p < 0.01). There were significant down-regulation of the protein expressions of caspase-3 and Bax (p < 0.01) in IPTS-treated H9c2 cells, as well as significant up-regulation of Bcl-2 protein expression (p < 0.01).Conclusion: These results suggest that IPTS can protect cardiomyocytes against apoptosis via the inhibition of oxidative stress and mitochondria-induced intrinsic apoptosis.Keywords: Ilex pubescens, Total saponins, Cardiomyocytes, Apoptosis, H9c2 cell

Effect of siRNA interference on nerve growth factor in intervertebral disc inflammation rats

AbstractObjectiveTo investigate the inhibition effect of siRNA interference on NGF induced by inflammatory factor IL-6, and IL-1 so as to provide novel targets for clinical treatment of discogenic low back pain.MethodsThe intervertebral disc nucleus and annulus fibrosus cells of rats were separated. The cells were co-cultured with different concentrations (10 nmol/L, 20 nmol/L, 50 nmol/L, 100 nmol/L) of IL-6 and IL-1β. The NGF-siRNA was leaded into the co-cultured cells with its import ability assessed by flow cytometry instrument tests, before and after which the NGF mRNA expression was detected by real-time Q-PCR and the NGF content was detected by ELISA.ResultsFlow cytometry instrument test results showed that the NGF-siRNA cell conversion rate was 99.8%. Real-time Q-PCR detection results showed that compared with negative control group, the NGF mRNA expression of co-cultured cells treated by 10 nmol/L, 20 nmol/L, 50 nmol/L, 100 nmol/L IL-6 and IL-1β were respectively raised 3.4, 3.7, 4.7, 3.7 times which were all significantly down-regulated after the import of NGF-siRNA. EILSA detection results showed that compared with negative control group, the NGF content of co-cultured medium treated by 10 nmol/L, 20 nmol/L, 50 nmol/L, 100 nmol/L I-L6 and IL-1β were respectively raised 2.9, 3.3, 4.5, 7.4 times which were all significantly decreased after the import of NGF-siRNA.ConclusionsThese molecular biological results suggest that inflammatory factor IL-6 and IL-1β could stimulate NGF on intervertebral disc cells in vitro culture model and its efficiency is concentration dependent, while siRNA interference can inhibit the stimulation effect of IL-6 and IL-1β on intervertebral disc cell, which provides a new targets for the clinical treatment of discogenic low back pain

Feline umbilical cord-derived mesenchymal stem cells: isolation, identification, and antioxidative stress role through NF-κB signaling pathway

At present, the differentiation potential and antioxidant activity of feline umbilical cord-derived mesenchymal stem cells (UC-MSCs) have not been clearly studied. In this study, feline UC-MSCs were isolated by tissue adhesion method, identified by flow cytometry detection of cell surface markers (CD44, CD90, CD34, and CD45), and induced differentiation toward osteogenesis and adipogenesis in vitro. Furthermore, the oxidative stress model was established with hydrogen peroxide (H2O2) (100 μM, 300 μM, 500 μM, 700 μM, and 900 μM). The antioxidant properties of feline UC-MSCs and feline fibroblasts were compared by morphological observation, ROS detection, cell viability via CCK-8 assay, as well as oxidative and antioxidative parameters via ELISA. The mRNA expression of genes related to NF-κB pathway was detected via quantitative real-time polymerase chain reaction, while the levels of NF-κB signaling cascade-related proteins were determined via Western Blot. The results showed that feline UC-MSCs highly expressed CD44 and CD90, while negative for CD34 and CD45 expression. Feline UC-MSCs cultured under osteogenic and adipogenic conditions showed good differentiation capacity. After being exposed to different concentrations of H2O2 for eight hours, feline UC-MSCs exhibited the significantly higher survival rate than feline fibroblasts. A certain concentration of H2O2 could up-regulate the activities of SOD2 and GSH-Px in feline UC-MSCs. The expression levels of p50, MnSOD, and FHC mRNA in feline UC-MSCs stimulated by 300 μM and 500 μM H2O2 significantly increased compared with the control group. Furthermore, it was observed that 500 μM H2O2 significantly enhanced the protein levels of p-IκB, IκB, p-p50, p50, MnSOD, and FHC, which could be reversed by BAY 11-7,082, a NF-κB signaling pathway inhibitor. In conclusion, it was confirmed that feline UC-MSCs, with good osteogenesis and adipogenesis abilities, had better antioxidant property which might be related to NF-κB signaling pathway. This study lays a foundation for the further application of feline UC-MSCs in treating the various inflammatory and oxidative injury diseases of pets

Australobius polyspinipes sp. n., a new species of Australobius Chamberlin, 1920 (Lithobiomorpha: Lithobiidae) from China

Australobius polyspinipes sp. n. (Lithobiomorpha: Lithobiidae) was recently discovered from Tianheshan Mountain, Hebei Province, China, and it is described here. Morphologically the new species is similar to A. nodulus Ma, Song &Zhu, 2008 and A. magnus (Trozina, 1894), both recorded from China. The new species can be easily distinguished from those by having 7+7–8+8 coxosternal teeth, 10–12 ocelli on each side of the cephalic plate, 5+5 spurs on the first article of the female gonopods and differences in plectrotaxy of legs. The main morphological characters and a key to the known Chinese species of genus Australobius based on adult specimens is presented

GL-CLeF: A Global-Local Contrastive Learning Framework for Cross-lingual Spoken Language Understanding

Due to high data demands of current methods, attention to zero-shot cross-lingual spoken language understanding (SLU) has grown, as such approaches greatly reduce human annotation effort. However, existing models solely rely on shared parameters, which can only perform implicit alignment across languages. We present Global--Local Contrastive Learning Framework (GL-CLeF) to address this shortcoming. Specifically, we employ contrastive learning, leveraging bilingual dictionaries to construct multilingual views of the same utterance, then encourage their representations to be more similar than negative example pairs, which achieves to explicitly aligned representations of similar sentences across languages. In addition, a key step in GL-CLeF is a proposed Local and Global component, which achieves a fine-grained cross-lingual transfer (i.e., sentence-level Local intent transfer, token-level Local slot transfer, and semantic-level Global transfer across intent and slot). Experiments on MultiATIS++ show that GL-CLeF achieves the best performance and successfully pulls representations of similar sentences across languages closer.Comment: Accepted at ACL2022 Main Conferenc

Detection and identification of NAP-2 as a biomarker in hepatitis B-related hepatocellular carcinoma by proteomic approach

<p>Abstract</p> <p>Background</p> <p><b>A </b>lack of sensitive and specific biomarkers is a major reason for the high rate of Primary hepatocellular carcinoma (HCC)-related mortality. The aim of this study was to investigate potential proteomic biomarkers specific for HCC.</p> <p>Methods</p> <p>81 patients with hepatitis B-related HCC and 33 healthy controls were randomly divided into a training set (33 HCC, 33 controls) and a testing set (48 HCC, 33 controls). Serum proteomic profiles were measured using Surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy (SELDI-TOF-MS).) A classification tree was established by Biomarker Pattern Software (BPS). Candidate SELDI peaks were isolated by tricine-SDS-PAGE, identified by HPLC-MS/MS and validated by immunohistochemistry (IHC) in liver tissues.</p> <p>Results</p> <p>A total of 6 proteomic peaks (3157.33 m/z, 4177.02 m/z, 4284.79 m/z, 4300.80 m/z, 7789.87 m/z, and 7984.14 m/z) were chosen by BPS to establish a classification tree with the highest discriminatory power in the training set. The sensitivity and specificity of this classification tree were 95.92%, and 100% respectively in the testing set. A candidate marker of about 7984 m/z was isolated and identified as neutrophil-activating peptide 2 (NAP-2). IHC staining showed that NAP-2 signals were positive in HCC tissues but negative in adjacent tissues.</p> <p>Conclusion</p> <p>The NAP-2 may be a specific proteomic biomarker of hepatitis B-related HCC.</p