Mediated Regulation Research Articles

Online mediation: international experience and prospects for implementation in Ukraine

The article discusses the problems of online mediation in Ukraine. The article analyzes the legislative regulation of mediation. As well as the use of online methods for the lawsuits resolution, which can significantly reduce the burden on the judicial system and ensure access to justice for all citizens under martial law and pandemic conditions (COVID-19). In the context of the pandemic, the introduction of online mediation becomes especially relevant, as it aims not only to simplify the process of lawsuits resolution, but also to ensure the preservation of citizens’ health. «The guidelines for preparing and conducting online mediation" adopted by the CA "NAMU" were analyzed. Created by the professional community of mediators as a response to the challenges that arise in the daily legal professional activities of mediators, such as: new circumstances, lack of legislation, etc. And distributed in order to provide support for safe professional practice. Determined the main factors of the prospects for the rapid development of alternative lawsuits resolution methods under digitalization and martial law in Ukraine.
 An analysis of the international legal regulation of online mediation has been carried out. It has been researched that the European legal system is aimed at encouraging the parties in lawsuits to use proper legal procedures to resolve the legal dispute, but not only in court. An analysis of the challenges of online mediation, which the mediator and participants face when conducting mediation in this format, has been carried out, as well as the advantages and disadvantages of online mediation in Ukraine have been determined. The ethical principles for online lawsuits resolution are set to find a wider audience as stakeholders in ODR systems become more expansive and inclusive. Different ways to improve the legislative regulation of online mediation by making additions to the Law of Ukraine "Mediation" are proposed.

A pan-cancer analysis of DDR1 in prognostic signature and tumor immunity, drug resistance

Disk-like domain receptor 1 (DDR1) is a crucial regulator of pro-inflammatory mediators and matrix-degrading enzymes. Although mounting evidence supports a vital role for DDR1 in the tumorigenesis of some cancers, no pan-cancer analysis of DDR1 has been reported. Therefore, we aimed to explore the prognostic value of DDR1 in 33 cancer types and investigate its potential immune function. We used a range of bioinformatics approaches to explore the potential carcinogenic role of DDR1 in multiple cancers. We found that DDR1 was expressed at high levels in most cancers. DDR1 expression was positively or negatively associated with prognosis in different cancers. DDR1 expression was significantly associated with DNA methylation in 8 cancers, while there was a correlation between DDR1 expression and RNA methylation-related genes and mismatch repair gene in most cancers. Furthermore, DDR1 expression was significantly associated with microsatellite instability in 6 cancers and tumor mutation burden in 11 cancers. In addition, DDR1 expression was also significantly correlated with immune cell infiltration, tumor microenvironment, immune-related genes, and drug resistance in various cancers. In conclusion, DDR1 can serve as a potential therapeutic target and prognostic marker for various malignancies due to its vital role in tumorigenesis and tumor immunity.

Abstract 2606: Requirement of MDM2 E3 ligase activity for regulating p53 during normal development, cell cycle regulation and genome integrity

Abstract p53 tumor suppressor, the most mutated gene in human cancer, is negatively regulated by MDM2 and MDM4 proteins. Two distinct mechanisms of MDM2/MDM4 regulation of p53 include, one, suppression of p53 transactivation activity by direct interaction of MDM2/MDM4 with p53 protein, thus blocking its transactivation interface and two, targeting p53 to proteasomal degradation by MDM2 E3 ligase activity. However, the importance of these mechanisms in normal p53 regulation in vivo has remained controversial. To genetically separate these two mechanisms of p53 regulation we generated a novel Mdm2 mutant mouse (Mdm2L466A) that lacks E3 ligase activity but retains the ability of Mdm2 to heterodimerize with Mdm4 and inhibit p53 transactivation. Homozygous Mdm2L466A mice are embryonic lethal due to dysregulated p53 activity, thus demonstrating Mdm2 E3 ligase mediated regulation of p53 is essential during embryonic development. Additionally, we uncovered novel p53-independent functions of MDM2 E3 ligase in cell cycle regulation and genome integrity in cells. Cells lacking MDM2 E3 ligase activity have defective G2-M transition during cell cycle and developed elevated levels of aneuploidy regardless of p53 status. This study unequivocally demonstrates the requirement of Mdm2 E3 ligase activity for normal regulation of p53 and uncovers previously unknown p53-independent role of Mdm2 in cell cycle regulation and maintaining genome integrity. A current therapeutic approach involves blocking MDM2-p53 interaction to promote p53 tumor suppression function in cancer cells. Our findings suggest that this approach alone is insufficient because MDM2 has other potential oncogenic mechanisms independent of its role as negative regulator of p53. Hence, targeting MDM2 E3 ligase activity is likely to be more effective cancer therapy as it addresses both p53-dependent and p53-independent oncogenic mechanisms of MDM2. Citation Format: Meenalakshmi Chinnam, Rati Lama, Chao Xu, Xiaojing Zhang, Carlos Cedeno, Yanqing Wang, Aimee B. Stablewski, David W. Goodrich, Xinjiang Wang. Requirement of MDM2 E3 ligase activity for regulating p53 during normal development, cell cycle regulation and genome integrity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2606.

Abstract 4908: Study on Ca 2+-mediated regulation mechanisms on chemotherapy-induced peripheral neuropathy in multiple myeloma and non-Hodgkin lymphoma

Abstract In general, when treating cancer patients, various anticancer drugs are used in combination to enhance the anticancer effect. However, most drugs exhibit side effects. Neurotoxicity, one of the side effects commonly accompanied, greatly affects the quality of life and also reduces compliance with treatment. Calcium signal transduction, which occupies a key position in cell signal transduction, has been found to be involved in various mechanisms including proliferation, differentiation, and death of cells, beyond the existing research direction that focuses on channels. To identify the mechanisms of calcium signal regulation that can reduce chemotherapy induced peripheral neuropathy (CIPN) while maintaining anticancer functions of drugs, we study with the two drugs frequently used in non-Hodgkin lymphoma and multiple myeloma, borteozomib (BTZ) and vincristine (VIN), whose main side effects are neurotoxicity. We used the human neuroblastoma cell line, SH-SY5Y cells, and performed MTT assay after treating them with N-Acetylcysteine ​​(NAC), an antioxidant, to determine whether the reduction in cell viability by BTZ and VIN was due to the increase in ROS. When BTZ and VIN were treated with NAC, cytotoxicity by BTZ was significantly reduced and cell viability decreased by VIN was significantly increased. From these results, we found that ROS are involved in BTZ and VIN -induced apoptosis. In addition, we performed MTT assay using calcium inhibitors to investigate whether intracellular calcium signaling is involved in BTZ and VIN-induced apoptosis, and 8-Br-cADPR, Ned-19, Xestospongin C, and Heparin were used as calcium inhibitors. To confirm that the cell viability reduced by BTZ and VIN is recovered by calcium inhibitors, BTZ and VIN were treated together with calcium inhibitors, respectively. Cell viability was found to increase significantly. These results suggest that intracellular calcium signaling is involved in cell death induced by BTZ and VIN. The increase in [ca2+]i in the cytoplasm when treated with BTZ and VIN was measured using a fluorescence spectrophotometer. It was shown that the treatment with BTZ and VIN increased the calcium concentration in the cytoplasm within 1 minute. To confirm that the concentration of intracellular calcium was reduced by the calcium inhibitor, BTZ and calcium inhibitor were treated together, and it was confirmed that the intracellular calcium concentration increased by BTZ was decreased in a concentration-dependent manner at 2uM and 3uM of the calcium inhibitor. In conclusion, this experiment suggests that intracellular calcium signaling is involved in the BTZ and VIN’s reduction of neuronal cell viability (CIPN) in multiple myeloma and non-Hodgkin lymphoma. This research was supported by NRF 2021R1A2C1091322 (J.-S.Kim) and 2021R1F1A1064120 (J. Park). Citation Format: Jinny Park, Jong-Suk Kim, Kwang-Hyun Park. Study on Ca 2+-mediated regulation mechanisms on chemotherapy-induced peripheral neuropathy in multiple myeloma and non-Hodgkin lymphoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4908.

Abstract 3642: SPHK1 mediated regulation of antitumor immunity in ovarian cancer

Abstract Introduction: Sphingosine kinase 1 (SPHK1) is an enzyme that helps to synthesize biolipid sphingosine-1-phosphate (S1P). We showed extracellular vesicles facilitate an increased accumulation of S1P in the ovarian tumor microenvironment (TME). S1P enhances the exhaustion of CD8 T cells, thus downregulating anti-tumor immunity. Furthermore, we also found less ovarian tumor burden in SPHK1−/− mice compared to WT mice. Based on our preliminary data, we hypothesized that the SPHK1 depleted tumor microenvironment increases the infiltration of the immune cells and its activity downregulates the tumor progression. Methods: SPHK1 global knockout mice and WT C57BL/6 mice were used in the study. Tumor cells (ID8 P53−/− BRCA−/−) were injected, and tumor progression was studied. Immunophenotyping was performed by flow cytometry on ascites and tumor samples. Activation of CD8 T cells, B cells, and dentritic cells were studied by flow cytometry. Bulk RNA sequencing was performed on SPHK1−/− and WT tumors. Results: SPHK1−/− mice showed regression in tumor growth in multiple tumor models. Tumor tissues from SPHK1−/− mice enhanced infiltration of CD4 and CD8 T cells, B cells, and dentritic cells compared to WT mice. Furthermore, infiltrated immune cells were active in SPHK1−/− mice. Bulk RNA sequencing also showed enhanced immune response in SPHK1−/− tumors compared to WT tumors. Conclusion: SPHK1−/- environment decreased the tumor growth by enhancing the infiltration and activation of immune cells. We are currently focusing on identifying the immune cells which are critical players in this process. We will further implement these findings in human immune cells for clinical benefits for ovarian cancer patients. Citation Format: Prachi Gupta, Sudhir Kumar, Ishaque Pulikkal Kadamberi, Sonam Mittal, Jasmine George, Anjali Geethadevi, Pradeep Chaluvally-Raghavan, Sunila Pradeep. SPHK1 mediated regulation of antitumor immunity in ovarian cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3642.

Whole-transcriptome sequencing identifies key differentially expressed circRNAs/lncRNAs/miRNAs/mRNAs and linked ceRNA networks in adult degenerative scoliosis.

Adult degenerative scoliosis (ADS) is forecast to be a prevalent disabling condition in an aging society. Universally, its pathogenesis is perceived as intervertebral disc degeneration (IDD), however, a thought-provoking issue is why precisely a subset of patients with disc degeneration develop ADS. Exploring the diversities between common IDD and ADS would contribute to unraveling the etiological mechanisms of ADS. Therefore, we aimed to integrate the circRNA, lncRNA, miRNA, and mRNA expression profiles from normal adults (Normal), patients with lumbar disc herniation (LDH), and ADS by whole transcriptome sequencing, which identifies critical functional ncRNA and ceRNA networks and crosstalk between the various transcripts. The fresh whole blood samples (n = 3/group) were collected from ADS patients, LDH patients, and healthy volunteers (Normal group), which were examined for mRNA, miRNA, lncRNA, and circRNA expression and screened for differentially expressed (DE) ncRNAs. Then, Gene Ontology (GO) and KEGG analyses were performed for gene annotation and enrichment pathways on the DE RNAs, which were constructed as a lncRNA-miRNA-mRNA network. Eventually, DE RNAs were validated by qRT-PCR targeting disc nucleus pulposus (NP) tissue in ADS and LDH group (n = 10/group). Compared to the LDH group, we identified 3322 DE mRNAs, 221 DE lncRNAs, 20 DE miRNAs, and 15 DE circRNAs in the ADS. In contrast to Normal, 21 miRNAs and 19 circRNAs were differentially expressed in the ADS. The expression of multiple differentially expressed ncRNAs was confirmed by qRT-PCR analysis to be consistent with the sequencing results. In addition, GO, and KEGG analysis demonstrated that most DE mRNAs and ncRNAs target genes are involved in various biological processes, including Endocytosis, Apoptosis, Rap1 signaling pathway, Notch signaling pathway, and others. The constructed lncRNA-miRNA-mRNA co-expression network was primarily related to angiogenesis and regulation. By focusing on comparing asymmetric and symmetric disc degeneration, whole-transcriptome sequencing and bioinformatics analysis systematically screened for key ncRNAs in the development of ADS, which provided an abundance of valuable candidates for the elucidation of regulatory mechanisms. The DE ncRNAs and the lncRNA-miRNA-mRNA network are intrinsically involved in the regulation of mediator and angiogenesis, which may contribute to the insight into the pathogenesis of ADS.

Cotinus coggygria Scop. Attenuates Acetic Acid-Induced Colitis in Rats by Regulation of Inflammatory Mediators.

In traditional medicine, many medicinal plants are used in the treatment of various diseases caused by inflammation. The objective of the present study is to elucidate for the first time the effects of Cotinus coggygria (CC) ethanol extract (CCE) on colonic structure and inflammation of acetic acid-induced ulcerative colitis in rats. Colonic damage was assessed using disease activity index score, enzyme-linked immunosorbent assay, and hematoxylin-eosin staining. Also, in vitro antioxidant activity of CCE was investigated by ABTS methods. Total phytochemical content of CCE was measured spectroscopically. Acetic acid caused colonic damage according to disease activity index and macroscopic scoring. CCE significantly reversed these damages. While the levels of proinflammatory cytokines TNF-alpha, IL-1beta, IL-6, and TGF-1beta increased in tissue with UC, IL-10 level decreased. CCE increased inflammatory cytokine levels to values close to the sham group. At the same time, while markers indicating disease severity such as VEGF, COX-2, PGE2, and 8-OHdG indicated the disease in the colitis group, these values returned to normal with CCE. Histological research results support biochemical analysis. CCE exhibited significant antioxidant against ABTS radical. Also, CCE was found to have a high content of total polyphenolic compounds. These findings provide evidence that CCE might be benefit as a promising novel therapy in the treatment of UC in humans due to high polyphenol content and justify the use of CC in folkloric medicine for treatment of inflamed diseases.

Transforming growth factor- β mediated regulation of epigenome is required for epithelial to mesenchymal transition associated features in liver cancer cells

Hepatocellular carcinoma (HCC) frequently unfolds under an inflammatory condition, which is a hub for a plethora of cytokines. A better understanding of the cytokine functions and their contributions to disease development is key to design of future therapeutic strategies and reduction of global HCC burden. In this context, one of the major cytokines present in the HCC tumour milieu is the transforming growth factor-β (TGF-β). One of its classical functions involve facilitation of epithelial to mesenchymal transition (EMT), in tumour cells, promoting an invasive phenotype. In spite of its clinical relevance, the cellular events associated with TGF-β-induced EMT and its molecular regulation is poorly elucidated. Therefore, as part of this study, we treated HCC cells with TGF-β and characterized the cellular processes associated with EMT. Interestingly, EMT triggered by TGF-β was found to be associated with cytostasis and altered cellular metabolism. TGF-β resulted in down-regulation of cell cycle-associated transcripts, like Cyclin A2 (CCNA2), and metabolic genes, like Glutamic-oxaloacetic transaminase 1 (GOT1) through epigenetic silencing. An overall increase in total histone repressive mark (H3K27me3) associated with a specific enrichment of H3K27me3 at the upstream promoter region of CCNA2 and GOT1 was observed after TGF-β exposure, leading to their down-regulation. Importantly, TGF-β-downstream signalling mediator- SMAD and chromatin repressive complex member-enhancer of zeste homolog 2 (EZH2) were found to co-immunoprecipitate and were required for the above effects. Overall, our findings reflect that HCC cells undergoing EMT, attain cytostasis and modulate metabolic demands to efficiently facilitate the EMT differentiation switch, and these events are regulated at the epigenomic level through TGF-β-mediated signalling. Our results provide better understanding of cellular invasive features which can lead to development of novel therapeutic strategies.

Deciphering the complete human-monkeypox virus interactome: Identifying immune responses and potential drug targets.

Monkeypox virus (MPXV) is a dsDNA virus, belonging to Poxviridae family. The outbreak of monkeypox disease in humans is critical in European and Western countries, owing to its origin in African regions. The highest number of cases of the disease were found in the United States, followed by Spain and Brazil. Understanding the complete infection mechanism of diverse MPXV strains and their interaction with humans is important for therapeutic drug development, and to avoid any future epidemics. Using computational systems biology, we deciphered the genome-wide protein-protein interactions (PPIs) between 22 MPXV strains and human proteome. Based on phylogenomics and disease severity, 3 different strains of MPXV: Zaire-96-I-16, MPXV-UK_P2, and MPXV_USA_2022_MA001 were selected for comparative functional analysis of the proteins involved in the interactions. On an average, we predicted around 92,880 non-redundant PPIs between human and MPXV proteomes, involving 8014 host and 116 pathogen proteins from the 3 strains. The gene ontology (GO) enrichment analysis revealed 10,624 common GO terms in which the host proteins of 3 strains were highly enriched. These include significant GO terms such as platelet activation (GO:0030168), GABA-A receptor complex (GO:1902711), and metalloendopeptidase activity (GO:0004222). The host proteins were also significantly enriched in calcium signaling pathway (hsa04020), MAPK signaling pathway (hsa04010), and inflammatory mediator regulation of TRP channels (hsa04750). These significantly enriched GO terms and KEGG pathways are known to be implicated in immunomodulatory and therapeutic role in humans during viral infection. The protein hubs analysis revealed that most of the MPXV proteins form hubs with the protein kinases and AGC kinase C-terminal domains. Furthermore, subcellular localization revealed that most of the human proteins were localized in cytoplasm (29.22%) and nucleus (26.79%). A few drugs including Fostamatinib, Tamoxifen and others were identified as potential drug candidates against the monkeypox virus disease. This study reports the genome-scale PPIs elucidation in human-monkeypox virus pathosystem, thus facilitating the research community with functional insights into the monkeypox disease infection mechanism and augment the drug development.

Cullin 3-Mediated Regulation of Intracellular Iron Homeostasis Promotes Thymic Invariant NKT Cell Maturation.

The E3 ubiquitin ligase cullin 3 (Cul3) is critical for invariant NKT (iNKT) cell development, as iNKT cells lacking Cul3 accumulate in the immature developmental stages. However, the mechanisms by which Cul3 mediates iNKT cell development remain unknown. In this study, we investigated the role of Cul3 in both immature and mature thymic iNKT cells using a mouse model with a T cell-specific deletion of Cul3. We found that mature iNKT cells lacking Cul3 proliferated and died more than wild-type cells did. These cells also displayed increased glucose metabolism and autophagy. Interestingly, we found that tight regulation of iron homeostasis is critical for iNKT cell development. Without Cul3, mature iNKT cells harbored higher levels of cytosolic iron, a phenotype associated with increased cell death. Taken together, our data suggest that Cul3 promotes iNKT cell development partially through intracellular iron homeostasis.

Gut microbial and metabolic characterization of Atlantic salmon (Salmon salar) challenged with Aeromonas salmonicida

The bacterium Aeromonas salmonicida (A. salmonicida) is a pathogen that causes high mortality of Atlantic salmon (Salmon salar), which results in great economic losses to the aquaculture industry in various countries. In this study, Atlantic salmon were exposed to low (LI) and high concentration A. salmonicida bacterial suspension by intraperitoneal injection for 48 h. The results of 16S rRNA sequencing and untargeted metabolomics showed that A. salmonicida exposure changed the structure and metabolomics of the gut microbiota in Atlantic salmon. Compared with the control group, the relative abundance of Romboutsia, Blautia, Agathobacter, Parasutterella, Fusicatenibacter, Butyricicoccus, Dongia, Clostridium sensu stricto 1, and Bacteroides significantly increased in infection groups, and these changes were closely related to some differential metabolites identified by metabolome analysis. In infection groups, we found that taurocholic acid, taurochenodeoxycholic acid, 7-ketolithocholic acid were significantly up-regulation, and significant down-regulation of Vitamin C, vitamin A, indirubin, and hexanoic acid. These differential metabolites are mainly involved in bile secretion, biosynthesis of antibiotics, inflammatory mediator regulation of transient receptor potential channels, tryptophan metabolism, vitamin digestion and absorption, and the ornithine cycle. Our analysis of the connection between the gut microbiota and metabolism confirmed that the gut microbiota may affect Atlantic salmon resist infection by A. salmonicida by regulating metabolism.

Filamin A is overexpressed in non-alcoholic steatohepatitis and contributes to the progression of inflammation and fibrosis

Non-alcoholic steatohepatitis (NASH) is a chronic and progressive liver disease characterized by steatosis, inflammation, and fibrosis. Filamin A (FLNA), an actin-binding protein, is involved in various cell functions, including the regulation of immune cells and fibroblasts. However, its role in the development of NASH through inflammation and fibrogenesis is not fully understood. In this study, we found that FLNA expression was increased in liver tissues of patients with cirrhosis and mice with non-alcoholic fatty liver disease (NAFLD)/NASH and fibrosis. Immunofluorescence analysis showed that FLNA was primarily expressed in macrophages and hepatic stellate cells (HSCs). Knocking down of FLNA by specific shRNA in phorbol-12-myristate-13-acetate (PMA)-derived THP-1 macrophages reduced lipopolysaccharide (LPS)-stimulated inflammatory response. The decreased mRNA levels of inflammatory cytokines and chemokines and suppression of the STAT3 signaling were observed in FLNA-downregulated macrophages. In addition, knockdown of FLNA in immortalized human hepatic stellate cells (LX-2 cells) resulted in decreased mRNA levels of fibrotic cytokines and enzymes involved in collagen synthesis, as well as increased levels of metalloproteinases and pro-apoptotic proteins. Overall, these results suggest that FLNA may contribute to the pathogenesis of NASH through its role in the regulation of inflammatory and fibrotic mediators.

Cell-Type-Specific Gene Regulatory Networks of Pro-Inflammatory and Pro-Resolving Lipid Mediator Biosynthesis in the Immune System.

Lipid mediators are important regulators in inflammatory responses, and their biosynthetic pathways are targeted by commonly used anti-inflammatory drugs. Switching from pro-inflammatory lipid mediators (PIMs) to specialized pro-resolving (SPMs) is a critical step toward acute inflammation resolution and preventing chronic inflammation. Although the biosynthetic pathways and enzymes for PIMs and SPMs have now been largely identified, the actual transcriptional profiles underlying the immune cell type-specific transcriptional profiles of these mediators are still unknown. Using the Atlas of Inflammation Resolution, we created a large network of gene regulatory interactions linked to the biosynthesis of SPMs and PIMs. By mapping single-cell sequencing data, we identified cell type-specific gene regulatory networks of the lipid mediator biosynthesis. Using machine learning approaches combined with network features, we identified cell clusters of similar transcriptional regulation and demonstrated how specific immune cell activation affects PIM and SPM profiles. We found substantial differences in regulatory networks in related cells, accounting for network-based preprocessing in functional single-cell analyses. Our results not only provide further insight into the gene regulation of lipid mediators in the immune response but also shed light on the contribution of selected cell types in their biosynthesis.

Acute herpes zoster treated with surrounding fire needling combined with electroacupuncture at Jiājĭ (夹脊EX-B2): A randomized controlled trial: 火针围刺联合夹脊电针治疗急性期带状疱疹:随机对照试验

ObjectivesTo investigate the clinical efficacy of surrounding fire needling combined with electroacupuncture (EA) at Jiājĭ (夹脊EX-B2) on acute herpes zoster, and explore the potential mechanism of this combined treatment by measuring the changes of serum inflammatory cytokines and pain mediators. MethodsSixty patients with acute herpes zoster were randomly assigned to either acupuncture plus western medicine group or western medicine group. In the western medicine group, the patients took orally mecobalamin and ganciclovir for 14 consecutive days. In the acupuncture plus western medicine group, on the base of the western medicine, the surrounding fire needling (fire needling was operated swiftly toward the center of skin lesions.) combined with EA at EX-B2 was provided. Acupoints of EX-B2 were selected based on the nerve ganglion corresponding to the skin lesions, as well as two adjacent nerve ganglions(one level upwards and one downwards) to the lesions, and were stimulated with EA once daily for 14 days. The change of symptoms from the blister eruption to scabbing and decrustation were observed. The score of visual analogue scale (VAS), serum inflammatory cytokines (interleukin-1β, interleukin-6, tumor necrosis factor-α) and pain mediators (calcitonin gene related peptide, substance P) before and after treatment, the scores of Pittsburgh sleep quality index (PSQI) and Hamilton depression scale (HAMD) before and after treatment as well as at 1 month of follow-up, and the overall effective rate and the incidence of post-hepatic neuralgia at 1 month of follow-up were assessed and compared between two groups, respectively. ResultsAfter treatment completion, the time of blister relief, scabbing and decrustation in the acupuncture plus western medicine group was significantly shorter when compared with that of the western medicine group (all P < 0.05). After treatment, VAS score, PSQI score and HAMD score were all reduced significantly when compared with that before treatment within group (P < 0.05). These scores in the acupuncture plus western medicine were reduced significantly compared with the western medicine group at the same time point (all P < 0.05). After treatment, the incidence of post-hepatic neuralgia was 6.89%, lower than that in western medicine group (50.00%, P < 0.05). After treatment, the values of serum inflammatory cytokines (IL-1β, IL-6, TNT-a) and pain mediators (CGRP, SP) were reduced when compared with the those before treatment within group (P < 0.05), and the reduction in the acupuncture plus western medicine group was larger than that in the western medicine group (P < 0.05). The total effective rate was 96.67% in the acupuncture plus western medicine group, higher than western medicine group (86.67%, P < 0.05). ConclusionsSurrounding fire needling therapy combined with EA at EX-B2 on the base of western medicine obtained better therapeutic effect on acute herpes zoster when compared with western medicine. This combined treatment may effectively relieve pain and depressive emotions, improve sleep quality and reduce the incidence of post-hepatic neuralgia. The underlying mechanism may related to regulation of pain mediators and reduction of inflammatory cytokines.

Crosstalk of Transcriptional Regulators of Adaptive Immune System and microRNAs: An Insight into Differentiation and Development.

MicroRNAs (miRNAs), as small regulatory RNA molecules, are involved in gene expression at the post-transcriptional level. Hence, miRNAs contribute to gene regulation of various steps of different cell subsets' differentiation, maturation, and activation. The adaptive immune system arm, which exhibits the most specific immune responses, is also modulated by miRNAs. The generation and maturation of various T-cell subsets concomitant with B-cells is under precise regulation of miRNAs which function directly on the hallmark genes of each cell subset or indirectly through regulation of signaling pathway mediators and/or transcription factors involved in this maturation journey. In this review, we first discussed the origination process of common lymphocyte progenitors from hematopoietic stem cells, which further differentiate into various T-cell subsets under strict regulation of miRNAs and transcription factors. Subsequently, the differentiation of B-cells from common lymphocyte progenitors in bone marrow and periphery were discussed in association with a network of miRNAs and transcription factors.

Beclin-1 dependent autophagy improves renal outcomes following Unilateral Ureteral Obstruction (UUO) injury.

Interstitial Fibrosis and Tubular Atrophy (IFTA) is the most common cause of long-term graft failure following renal transplant. One of the hallmarks of IFTA is the development of interstitial fibrosis and loss of normal renal architecture. In this study, we evaluated the role of autophagy initiation factor Beclin-1 in protecting against post-renal injury fibrosis. Adult male wild type (WT) C57BL/6 mice were subjected to Unilateral Ureteral Obstruction (UUO), and kidney tissue samples were harvested at 72-hour, 1- and 3-week post-injury. The UUO-injured and uninjured kidney samples were examined histologically for fibrosis, autophagy flux, inflammation as well activation of the Integrated Stress Response (ISR). We compared WT mice with mice carrying a forced expression of constitutively active mutant form of Beclin-1, Becn1F121A/F121A . In all experiments, UUO injury induces a progressive development of fibrosis and inflammation. These pathological signs were diminished in Becn1F121A/F121A mice. In WT animals, UUO caused a strong blockage of autophagy flux, indicated by continuously increases in LC3II accompanied by an over 3-fold accumulation of p62 1-week post injury. However, increases in LC3II and unaffected p62 level by UUO were observed in Becn1F121A/F121A mice, suggesting an alleviation of disrupted autophagy. Beclin-1 F121A mutation causes a significant decrease in phosphorylation of inflammatory STING signal and limited production of IL6 and IFNγ, but had little effect on TNF-α, in response to UUO. Furthermore, activation of ISR signal cascade was detected in UUO-injured in kidneys, namely the phosphorylation signals of elF2S1 and PERK in addition to the stimulated expression of ISR effector ATF4. However, Becn1F121A/F121A mice did not reveal signs of elF2S1 and PERK activation under the same condition and had a dramatically reduced ATF level at 3-week post injury. The results suggest that UUO causes a insufficient, maladaptive renal autophagy, which triggered downstream activation of inflammatory STING pathway, production of cytokines, and pathological activation of ISR, eventually leading to the development of fibrosis. Enhancing autophagy via Beclin-1 improved renal outcomes with diminished fibrosis, via underlying mechanisms of differential regulation of inflammatory mediators and control of maladaptive ISR.

Salivary irisin level is higher and related with interleukin-6 in generalized periodontitis.

Irisin plays an important role in energy homeostasis, inflammation, glucose, and lipid metabolism, and it is shown to have relations with many inflammatory diseases. The aim of the study was to determine saliva and serum irisin and IL-6 levels in patients with stage III/grade B periodontitis compared with individuals with healthy periodontium. Twenty patients with stage III grade B periodontitis (P) and 20 periodontally healthy subjects (control; C) were included in this study. Clinical periodontal measurements were recorded. Saliva and serum levels of irisin and interleukin-6 (IL-6) were analyzed by enzyme-linked immunosorbent assay. Salivary irisin and IL-6 levels were significantly higher in the periodontitis group (p < 0.001, p = 0.002, respectively). Furthermore, serum IL-6 levels were found significantly higher in the periodontitis group compared with controls (p = 0.011). There was no significant difference between the periodontitis and control for serum irisin levels (p > 0.05). Significant positive correlations were found between all periodontal parameters and salivary irisin and IL-6 (p < 0.05) and also between BMI and saliva and serum IL-6 (respectively; r = 0.530, r = 0.329, p < 0.05). There was a positive correlation between salivary irisin and IL-6 (r = 0.369, p < 0.05). Monitoring of salivary irisin and IL-6 might be potential biomarker for predicting the susceptibility to periodontitis. Scientific rationale for the study: Irisin is a novel adipomyokine that has played an important role in energy homeostasis, glucose and lipid metabolism, angiogenesis, immunity, and inflammation. Irisin is involved in the pathogenesis of diseases affecting many body systems. IL-6, another adipomyokine, is a major inflammatory mediator and homeostatic regulator of glucose and lipid metabolism and is associated with periodontitis. No studies investigated the relationship between advanced periodontal disease, irisin, and IL-6 together. The salivary irisin and IL-6 levels were significantly higher and positively correlated in patients with periodontitis relative to healthy controls. Furthermore, serum IL-6 levels were significantly increased in patients with periodontitis. The study shows that irisin and IL-6 can be candidate salivary biomarkers for periodontitis and predict to periodontal status.

N-Acetylcysteine Mediated Regulation of MnSOD, UCP-2 and Cytochrome C Associated with Amelioration of Monocrotophos-Induced Hepatotoxicity in Rats

Pesticides are now a risk to the environment and public health. Monocrotophos (MCP) is known to cause organ toxicity and impart degenerative effects at cellular levels. N-acetylcysteine (NAC) is a natural antioxidant having various prophylactic properties. Male Wistar rats were given NAC (200 mg/kg b.wt), MCP (0.9 mg/kg b.wt) and NAC followed by MCP; intragastrically for 28 consecutive days. Regulation of MnSOD, UCP-2 and cytochrome c was analyzed by western blotting and polymerase chain reaction. Histology, electron microscopy and weight parameters were evaluated in the liver. MCP exposure significantly decreased body weight gain, relative liver weight, and structural changes. Altered MnSOD protein expression, decreased transcription of UCP-2 and MnSOD, and released cytochrome c indicated that oxidative stress is involved in MCP exposure. Treatment of NAC to MCP-exposed rats normalized the weight and structural changes, restored MnSOD and UCP-2 levels and prevented the release of cytochrome c. The present study suggests that the regulation of UCP-2, MnSOD and cytochrome c is involved in NAC efficacy against MCP toxicity. These findings illustrate that NAC can serve as a potential therapeutic agent for toxicity and oxidative stress in mammals.

Differential expression profiling of tRNA-Derived small RNAs and their potential roles in methamphetamine self-administered rats.

Transfer RNA-derived small RNAs (tsRNAs) are a novel class of short, non-coding RNAs that are closely associated with the pathogenesis of various diseases. Accumulating evidence has demonstrated their critical functional roles as regulatory factors in gene expression regulation, protein translation regulation, regulation of various cellular activities, immune mediation, and response to stress. However, the underlying mechanisms by which tRFs & tiRNAs affect methamphetamine-induced pathophysiological processes are largely unknown. In this study, we used a combination of small RNA sequencing, quantitative reverse transcription-polymerase chain reaction (qRT‒PCR), bioinformatics, and luciferase reporter assays to screen the expression profiles and identify the functional roles of tRFs and tiRNAs in the nucleus accumbens (NAc) of methamphetamine self-administration rat models. A total of 461 tRFs & tiRNAs were identified in the NAc of rats after 14days of methamphetamine self-administration training. Of those, 132 tRFs & tiRNAs were significantly differentially expressed: 59 were significantly upregulated, whereas 73 were significantly downregulated in the rats with methamphetamine self-administration. Decreased expression levels of tiRNA-1-34-Lys-CTT-1 and tRF-1-32-Gly-GCC-2-M2, as well as increased expression levels of tRF-1-16-Ala-TGC-4 in the METH group compared with the saline control were validated by using RT‒PCR. Then, bioinformatic analysis was performed to analyse the possible biological functions of tRFs & tiRNAs in methamphetamine-induced pathogenesis. Furthermore, tRF-1-32-Gly-GCC-2-M2 was identified to target BDNF using the luciferase reporter assay. An altered tsRNA expression pattern was proven, and tRF-1-32-Gly-GCC-2-M2 was shown to be involved in methamphetamine-induced pathophysiologic processes by targeting BDNF. The current study provides new insights for future investigations to explore the mechanisms and therapeutic methods for methamphetamine addiction.

Specific Human Milk Oligosaccharides Differentially Promote Th1 and Regulatory Responses in a CpG-Activated Epithelial/Immune Cell Coculture.

Proper early life immune development creates a basis for a healthy and resilient immune system, which balances immune tolerance and activation. Deviations in neonatal immune maturation can have life-long effects, such as development of allergic diseases. Evidence suggests that human milk oligosaccharides (HMOS) possess immunomodulatory properties essential for neonatal immune maturation. To understand the immunomodulatory properties of enzymatic or bacterial produced HMOS, the effects of five HMOS (2'FL, 3FL, 3'SL, 6'SL and LNnT), present in human milk have been studied. A PBMC immune model, the IEC barrier model and IEC/PBMC transwell coculture models were used, representing critical steps in mucosal immune development. HMOS were applied to IEC cocultured with activated PBMC. In the presence of CpG, 2'FL and 3FL enhanced IFNγ (p < 0.01), IL10 (p < 0.0001) and galectin-9 (p < 0.001) secretion when added to IEC; 2'FL and 3FL decreased Th2 cell development while 3FL enhanced Treg polarization (p < 0.05). IEC were required for this 3FL mediated Treg polarization, which was not explained by epithelial-derived galectin-9, TGFβ nor retinoic acid secretion. The most pronounced immunomodulatory effects, linking to enhanced type 1 and regulatory mediator secretion, were observed for 2'FL and 3FL. Future studies are needed to further understand the complex interplay between HMO and early life mucosal immune development.