Corrigendum: Study on the pathogenesis of MiR-6324 regulating diarrheal irritable bowel syndrome and bioinformatics analysis.

[This corrects the article DOI: 10.3389/fphar.2023.1044330.].

Study on the pathogenesis of MiR-6324 regulating diarrheal irritable bowel syndrome and bioinformatics analysis.

Objective: To investigate the pathogenesis of IBS-D by bioinformatics analysis of the differential microRNAs in rat colon tissue and to analyze and predict the function of their target genes. Methods: Twenty male Wistar rats of SPF class were randomly divided into two groups, the model group was manipulated using the colorectal dilatation method + chronic restraint stress method to establish the IBS-D model; while the blank group stroked the perineum at the same frequency. Screening of differential miRNAs after High-throughput sequencing of rat colon tissue. GO and KEGG analysis of target genes using the DAVID website, further mapping using RStudio software; the STRING database and the Cytoscape software were used to obtain the protein interaction network (PPI) of the target genes as well as the core genes. Finally, qPCR was used to detect the expression of target genes in the colon tissue of two groups of rats. Results: After the screening, miR-6324 was obtained as the key of this study. The GO analysis of target genes of miR-6324 is mainly involved in protein phosphorylation, positive regulation of cell proliferation, and intracellular signal transduction; it affects a variety of cellular components such as cytoplasm, nucleus, and organelles on the intracellular surface; it is also involved in molecular functions such as protein binding, ATP binding, and DNA binding. KEGG analysis showed that the intersecting target genes were mainly enriched in cancer pathways, proteoglycans in cancer, neurotrophic signaling pathway, etc. The protein-protein interaction network screened out the core genes mainly Ube2k, Rnf41, Cblb, Nek2, Nde1, Cep131, Tgfb2, Qsox1, and Tmsb4x. The qPCR results showed that the expression of miR-6324 decreased in the model group, but the decrease was not significant. Conclusion: miR-6324 may be involved in the pathogenesis of IBS-D as a potential biological target and provide further ideas for research on the pathogenesis of the disease or treatment options.

Role of Blood Neurofilaments in the Prognosis of Amyotrophic Lateral Sclerosis: A Meta-Analysis.

Background: Neurofilaments in cerebrospinal fluid (CSF) and in blood are considered promising biomarkers of amyotrophic lateral sclerosis (ALS) because their levels can be significantly increased in patients with ALS. However, the roles of neurofilaments, especially blood neurofilaments, in the prognosis of ALS are inconsistent. We performed a meta-analysis to explore the prognostic roles of blood neurofilaments in ALS patients. Methods: We searched all relevant studies on the relationship between blood neurofilament levels and the prognosis of ALS patients in PubMed, Embase, Scopus, and Web of Science before February 2, 2021. The quality of the included articles was assessed using the Quality in Prognosis Studies (QUIPS) scale, and R (version 4.02) was used for statistical analysis. Results: Fourteen articles were selected, covering 1,619 ALS patients. The results showed that higher blood neurofilament light chain (NfL) levels in ALS patients were associated with a higher risk of death [medium vs. low NfL level: HR = 2.43, 95% CI (1.34-4.39), p < 0.01; high vs. low NfL level: HR = 4.51, 95% CI (2.45-8.32), p < 0.01]. There was a positive correlation between blood phosphorylated neurofilament heavy chain (pNfH) levels and risk of death in ALS patients [HR = 1.87, 95% CI (1.35-2.59), p < 0.01]. The levels of NfL and pNfH in blood positively correlated with disease progression rate (DPR) of ALS patients [NfL: summary r = 0.53, 95% CI (0.45-0.60), p < 0.01; pNfH: summary r = 0.51, 95% CI (0.24-0.71), p < 0.01]. Conclusion: The blood neurofilament levels can predict the prognosis of ALS patients; specifically, higher levels of blood neurofilaments are associated with a greater risk of death.

Chiral Fluorescent Metal-Organic Framework with a Pentanuclear Copper Cluster as an Efficient Luminescent Probe for Dy3+ Ion and Cyano Compounds.

Luminescent probes have been used for the detection of various heavy metals and toxic compounds. A novel sensor with excellent sensitivity and selectivity is in high demand. Herein, we designed and synthesized a three-dimensional copper-organic framework of "pcu" α-Po primitive cubic topology with a Schläfli symbol of {4.4.4.4.4.4.4.4.4.4.4.4.*.*.*}. By taking advantage of metal clusters and a triazole ligand as the metal-organic framework (MOF) components, the newly obtained MOF is stable in various environments and can be potentially used as the sensor. Remarkably, this MOF-based sensor shows high sensitivity and selectivity toward a dysprosium ion (Dy3+) in a multiple-lanthanide mixed solution. Besides, it exhibits luminescent quenching toward various cyano compounds. This chiral cluster-based network provides a potential luminescent probe for various inorganic and organic compounds with high sensitivity and selectivity.

[Comparison Between Different Mitochondrial Staining Methods in Cell and Tissue Samples].

OBJECTIVE: To compare the effects of mitochondria staining between specific mitochondrial fluorescent probes and anti-mitochondrial protein antibody in cell and tissue samples. METHODS: The HepG2 cells fixed by 4% paraformaldehyde were stained with MitoTracker Deep Red (100 nmol/L) or anti-Grp75 antibody (75 nmol/L or 100 nmol/L). The human healthy liver tissue samples fixed by 4% paraformaldehyde were stained with 150 nmol/L MitoTracker Deep Red or anti-Grp75 antibody. The above stained cell and tissue samples were observed using confocal microscopy. RESULTS: We found non-specific staining in HeLa cells and obscure mitochondrial image using MitoTracker Deep Red probes, while clear tubular and punctate distribution using anti-Grp75 antibody. In contrast, we observed more specific and better effects of MitoTracker Deep Red probes-stained liver tissue samples as compared to the antibody. CONCLUSION: To visualize mitochondria, the anti-Grp75 antibody staining worked better on cells and the MitoTracker Deep Red probes are more suitable for tissue samples.

[Expression and Significance of HIF-3α in Mitochondria].

OBJECTIVE: To investigate the mitochondrial translocation of hypoxia inducible factor-3α (HIF-3α) under normoxia and hypoxia and its physiological and pathological meanings. METHODS: ① After hypoxic (1%O2) or DMOG, CoCl2 treatments mimicking the hypoxic treatment, Western blot and immunofluorescence were used to examine the HIF-3α expression in mitochondria of HeLa and ACHN cells, respectively. ②The protease sensitivity experiment was used to explore the sub-organelle localization of HIF-3α in mitochondria. ③Western blot was used to examine mitochondrial HIF-3α in the normal mouse tissues and human liver carcinoma tissues. RESULTS: ① In HeLa and ACHN cells, HIF-3α translocated to mitochondria under normoxia and hypoxia, and its mitochondrial expression was higher under hypoxia; ②The protease sensitivity of HIF-3α was similar to proteins locating in the mitochondrial outer membrane; ③Mitochondrial HIF-3α expressed in multiple normal mouse tissues; The expression of mitochondrial HIF-3α was higher in human liver carcinoma tissues than the normal and adjacent tissues. CONCLUSIONS: HIF-3α translocated to mitochondrial outer membrane under both normoxia and hypoxia, and hypoxia could up-regulated HIF-3α mitochondrial translocation. Meanwhile, the phenomenon may be involved in the process of liver carcinoma.

Pharmacokinetics and tissue distribution of Ebracteolatain A, a potential anti-cancer compound, as determined by an optimized ultra-performance liquid chromatography tandem mass spectrometry method.

Ebracteolatain A is a phloroglucinol derivative from the root of Euphorbia ebracteolata Hayata, a Traditional Chinese Medicine also known as Langdu. It has been shown to have good inhibitory effects in breast cancer cells. In this study, a simple, rapid, sensitive, and specific ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to study the pharmacokinetics (PKs) and tissue distribution of Ebracteolatain A in rats. Ebracteolatain A and Magnolol (internal standard) were extracted by the simple protein precipitation extraction technique using methanol as the precipitating solvent. Chromatographic separation was performed using the Agilent Poroshell 120 EC-C18 column with a mobile phase of acetonitrile:0.1% formic acid (70:30, v/v). The protonated analyte was quantitated in negative ionization by MS/MS via multiple reaction monitoring mode. The assay exhibited a linear dynamic range of 2-2000 ng/mL for Ebracteolatain A in biological samples. The lower limit of quantitation was 2 ng/mL. Non-compartmental PK parameters indicated that Ebracteolatain A was well absorbed into the systemic circulation. The absolute bioavailability of Ebracteolatain A was greater when administered by intraperitoneal administration than by oral administration. The tissue distribution study showed that Ebracteolatain A was distributed in the heart, liver, spleen, lung, kidney, brain, stomach, intestine, uterus, ovary, and breast after intravenous injection. The results of this study further our understanding of the in vivo anti-cancer activity of Ebracteolatain A, and shed light on pharmacological strategies that may be useful for the development of novel breast cancer therapeutics.

HIF-1α protects against oxidative stress by directly targeting mitochondria.

The transcription factor hypoxia inducible factor-1α (HIF-1α) mediates adaptive responses to oxidative stress by nuclear translocation and regulation of gene expression. Mitochondrial changes are critical for the adaptive response to oxidative stress. However, the transcriptional and non-transcriptional mechanisms by which HIF-1α regulates mitochondria in response to oxidative stress are poorly understood. Here, we examined the subcellular localization of HIF-1α in human cells and identified a small fraction of HIF-1α that translocated to the mitochondria after exposure to hypoxia or H2O2 treatment. Moreover, the livers of mice with CCl4-induced fibrosis showed a progressive increase in HIF-1α association with the mitochondria, indicating the clinical relevance of this finding. To probe the function of this HIF-1α population, we ectopically expressed a mitochondrial-targeted form of HIF-1α (mito-HIF-1α). Expression of mito-HIF-1α was sufficient to attenuate apoptosis induced by exposure to hypoxia or H2O2-induced oxidative stress. Moreover, mito-HIF-1α expression reduced the production of reactive oxygen species, the collapse of mitochondrial membrane potential, and the expression of mitochondrial DNA-encoded mRNA in response to hypoxia or H2O2 treatment independently of nuclear pathways. These data suggested that mitochondrial HIF-1α protects against oxidative stress induced-apoptosis independently of its well-known role as a transcription factor.

Adoptive transfusion of tolerogenic dendritic cells prolongs the survival of liver allograft: a systematic review.

OBJECTIVES: To systematically review the effects of tolerogenic dendritic cells (Tol-DCs) induced by different methods on liver transplantation and their possible mechanisms of action. METHODS: PubMed and EMbase were searched for relevant articles through 31 December 2013. The effects of Tol-DCs on liver allograft survival were semiquantitatively evaluated, and the possible mechanisms by which Tol-DCs prolong graft survival were analyzed. RESULTS: Seven articles were included, and classified according to methods of induction, sources, and methods of infusing Tol-DCs. Tol-DCs induced from immature DCs (imDCs), with cytokines, and by gene modification induced liver transplant tolerance for 33.1 ± 32.5 days (2.7-fold vs. control), 26.17 ± 16.20 days (1.8-fold vs. control), and 11.7 ± 1.6 days (2.3-fold vs. control), respectively. DCs derived from recipient bone marrow, donor bone marrow, and donor spleen induced liver transplant tolerance for 51.0 ± 0.0 days (5.9-fold vs. control), 21.4 ± 26.8 days (2.4-fold vs. control), and 15.0 ± 0.0 days (2.3-fold vs. control), respectively. The primary mechanisms by which Tol-DCs induce liver transplant tolerance were the induction of T-cell hyporeactivity and Th2 differentiation. CONCLUSIONS: Tol-DCs induced by three different methods could extend liver allograft survival, with imDCs showing optimal results. The optimal infusion method was intravenous injection of 1-2 × 10(6) Tol-DC, similar to findings in renal transplantation. Tol-DCs prolonged liver transplant tolerance more than renal transplant tolerance.

Adoptive transfusion of tolerant dendritic cells prolong the survival of renal allografts: a systematic review.

OBJECTIVE: The aim of this study was to systematically review the effects of transfusing Tol-DCs induced by different methods on renal transplantation and survival time. METHOD: PubMed and EMbase were searched for relevant articles from inception to July 20(th), 2013. Renal allograft survival time was regarded as the endpoint outcome. The effects of Tol-DCs on renal transplantation were evaluated semi-quantitatively. RESULTS: Sixteen articles were included. There were three sources of Tol-DCs, including bone marrow, spleen, and thoracic duct lymph node. Rats were administrated cells intravenously and 83% of mice through the portal vein. Four subtypes of bone marrow Tol-DCs enhanced renal allograft time: immature DCs enhanced allograft survival 4.9-fold in rats and 2.0-fold in mice, gene modified DCs enhanced allograft survival 4.4-fold in rats and 2.2-fold in mice, and drug and cytokine induced enhanced allograft survival 2.9-fold and 2.7-fold, respectively, in rats. Tol-DCs from the spleen and thoracic duct lymph nodes prolonged allograft survival 2.7-fold and 1.8-fold, respectively, in rats. 1-2 × 10(6) doses of Tol-DCs extended the survival time of rats following renal transplantation. The key mechanisms by which Tol-DCs enhance allograft and overall survival included: (i) inducing T-cell hyporeactivity; (ii) reducing the effects of cytotoxic lymphocytes; and (iii) inducing Th2 differentiation. CONCLUSION: Bone marrow Tol-DCs can extend allograft survival and induce immune tolerance in fully MHC-mismatched renal transplantation in rats and mice. The effects of imDCs and gene modified Tol-DCs in mice are less marked. In conclusion, a single-injection of 1-2 × 10(6) doses of bone marrow Tol-DCs (i.v.), in combination with an immune-suppressor, a co-stimulator, and accessory cells can significantly extend renal allograft survival.

[Rapid identification of chemical constituents in Salvia chinensis by HPLC-TOF-MS].

It's established a high-performance liquid chromatography-time of flight mass spectrometry(HPLC-TOF-MS) method to analyze chemical constituents in Salvia chinensis. The separation was performed on a SHISEIDO MG C18 reverse phase column (3.0 mm x 100 mm, 3 microm). The mobile phase consisted of acetonitrile (A) and water (containing 0.1% formic acid, B) was used as gradient elute. The gradient of a phase, 10%-90% (0-33 min), 90% (33-40 min). The flow rate was 0.6 mL x min(-1). Post-column split ratio was 2:1. Temperature of column was 25 degrees C. Time-of-flight mass spectrometer (TOF-MS) and electrospray ion source (ESI) was applied for qualitative analysis under positive ion mode, and mass scan range was m/z 100-1 000. As a result,28 of the major chemical constituents of S. chinensis were identified by HPLC-TOF-MS. In this study, a rapid and efficient method for studying the chemical constituents in S. chinensis by HPLC-TOF-MS was established, which paves a way for the quality control and further studies of the herb in vivo.

[Experiment on inducing human dental pulp stem cells into neural-like cells].

OBJECTIVE: To explore the multi-differentiated capability of human dental pulp stem cells (hDPSCs) obtained by cell-clone culture approach and to determine the appropriate induced medium. METHODS: The cloned isolation and expansion of hDPSCs were preinduced for 24 h, and were subsequently replaced with neural-inductive medium containing certain concentration of dimethylsulfoxide (DMSO), butylated hydroxyanisode (BHA), forskolin, P-mercaptoethanol (p-ME) and hydrocortisone for 4 days. Then induced cells were analyzed by morphological observation, immnocytochemical staining for non-specific esterase (NSE) and glial fibrillary acidic protein (GFAP) expression, RT-PCR for GFAP mRNA. Meanwhile, the uninduced hDPSCs were used as negative control. RESULTS: The morphology of induced cells changed at the initial 12 h, and displayed a typical neuron-like cells at 24 h. There was a gradual increase in the number of these neuronal differentiated cells with continuous induction. Furthermore, immnocytochemical staining showed that the induced cell expressed NSE and GFAP, two marked enzymes of neuron cell. The GFAP mRNA was also detected in induced cells by RT-PCR assay. In contrast, the uninduced cells maintained its original appearance and had no expression on them. CONCLUSION: hDPSCs may possess potential of multiple-differentiation and may differentiate into neuron-like cells on certain inductive condition.