The nuclear factor-κB (NF-κB) family is evolutionary conserved and plays key roles in the regulation of numerous basic cellular processes. In this study, a sea cucumber Holothuria leucospilota NF-κB1 p105 named HLp105 was first obtained. The full-length cDNA of HLp105 is 6564 bp long, with a 219 bp 5' untranslated region (UTR), a 2979 bp 3' UTR, and a 3366 bp open reading frame (ORF) encoding for 1121 amino acids with a deduced molecular weight of 123.92 kDa and an estimated pI of 5.31. HLp105 protein contains the conserved domain RHD, IPT, ANK and DEATH. HLp105 mRNA can be detected in all tissues examined, with the highest level in the intestine, followed by the transverse vessel, rete mirabile, coelomocytes, respiratory tree, bolishiti, cuvierian tubules, body wall, oesophagus and muscle. Challenged by LPS or poly (I:C), the transcription level of HLp105 was apparently up-regulated in the tissues examined. Besides, Over-expression of HLp105 in HEK293T cells, the apoptosis was inhibited, and the cytokines IL-1β and TNF-α were activated. The results are important for better understanding the function of NF-κB1 p105 in sea cucumber and reveal its involvement in immunoreaction.

Keywords: Holothuria leucospilota; NF-κB1 p105; apoptosis; expression patterns.

Following the publication of this article [1], it was noted that the author list was incomplete and was missing the following author.

UNASSIGNED

In the human evolutionary history, Single Nucleotide Polymorphism (SNP) frequencies are valuable in terms of finding connections between different populations. Due to the pronounced role of the immune system in combating pathogens and environmental stressors, polymorphisms in the immune genes are subject to selection pressure of the diseases as well. The functional polymorphisms in NF-κB1 promoter (-94 ins/del) are associated with different diseases; therefore, we aimed to establish the frequencies of NF-κB1 rs28362491 alleles in a population of Southwestern Iranians in comparison with the world populations.

UNASSIGNED

We assessed the polymorphism of -94 ATTG ins/del (rs28362491) in 201 Iranian healthy blood donors from Fars Province, central Iran in a one year period between 2015 and 2016 by PCR-RFLP method using DNA extracted from peripheral blood mononuclear cells.

UNASSIGNED

The frequency of ins/ins homozygote genotype was found to be 46.97%. The frequency of heterozygote individuals was 42.42% and the percentage of del/del homozygote genotype was 10.61%. We observed a genetic similarity based on the genotype frequencies of NF-κB1 -94 ins/del ATTG polymorphism between our sample of Iranians with American Jewish, Turkish, American non-Jewish, Chinese-Uyghurs and Germans.

UNASSIGNED

The results confirmed genetic interrelation of Iranians with some ancient neighbors and their admixture with countries along the Silk Road. We suggest that mapping the distribution of NF-κB1-94 ATTG ins/del along with HLA genes may help to better define the relations between human populations and design population-specific vaccines for pathogens with a high rate of variation.

Context It has been found that hydroxysafflor-yellow A (HSYA) inhibits angiogenesis and the proliferation of abnormal human umbilical vein endothelial cells (HUVECs) in our previous study; however, the mechanism is still unclear. Objective This study investigates the mechanisms of HSYA inhibiting abnormal proliferation of HUVECs through detecting the expression of vascular endothelial growth factor (VEGF) and its receptor (KDR), and the protein expression in the Ras-Raf-MEK-ERK-signalling pathway. Materials and methods HepG2 cell cultural supernatant was used to culture HUVECs to make promote abnormal proliferation, and HSYA was added into the medium. The expression of VEGF, KDR, c-myc, N-ras and NF-κB1 in abnormal HUVEC was detected by RT-qPCR and ELISA at the mRNA and protein levels. Protein expression of ERK signal pathway was measured by Western blot. Results Compared with the abnormal proliferation of HUVECs without any treatment, HSYA inhibited the expression of VEGF and KDR in vitro. Similarly, the protein expression of Ras, p-raf, p-ERK and p-p38MARK in the abnormal HUVECs was reduced when they were treated by HSYA, especially in p-ERK, yet the total raf, ERK, p38MAPK and Akt were not changed whether HSYA existed or not. HSYA could also inhibit the expression of c-myc, N-ras, and NF-κB1. Conclusion When the abnormal HUVECs were treated with HSYA, the low expression of VEGF and KDR reduced the expression of oncogene and transcription factor through the Ras-Raf-MEK-ERK1/2 pathway of the MAPK family. This resulted in inhibiting the abnormal proliferation of HUVECs and angiogenesis.

The MKK1/2 kinase tumour progression locus 2 (TPL-2) is critical for the production of tumour necrosis factor alpha (TNFα) in innate immune responses and a potential anti-inflammatory drug target. Several earlier pharmaceutical company screens with the isolated TPL-2 kinase domain have identified small-molecule inhibitors that specifically block TPL-2 signalling in cells, but none of these have progressed to clinical development. We have previously shown that TPL-2 catalytic activity regulates TNF production by macrophages while associated with NF-κB1 p105 and ABIN-2, independently of MKK1/2 phosphorylation via an unknown downstream substrate. In the present study, we used a positional scanning peptide library to determine the optimal substrate specificity of a complex of TPL-2, NF-κB1 p105 and ABIN-2. Using an optimal peptide substrate based on this screen and a high-throughput mass spectrometry assay to monitor kinase activity, we found that the TPL-2 complex has significantly altered sensitivities versus existing ATP-competitive TPL-2 inhibitors than the isolated TPL-2 kinase domain. These results imply that screens with the more physiologically relevant TPL-2/NF-κB1 p105/ABIN-2 complex have the potential to deliver novel TPL-2 chemical series; both ATP-competitive and allosteric inhibitors could emerge with significantly improved prospects for development as anti-inflammatory drugs.

Mesenchymal stromal cells (MSCs) can potently regulate the functions of immune cells and are being investigated for the management of inflammatory diseases. Toll-like receptor 3 (TLR3)-stimulated human MSCs (hMSCs) exhibit increased migration and chemotaxis within and toward damaged tissues. However, the regulatory mechanisms underlying these migratory activities are unclear. Therefore, we analyzed the migration capability and gene expression profiles of TLR3-stimulated hMSCs using RNA-Seq, wound healing, and transwell cell migration assay. Along with increased cell migration, the TLR3 stimulation also increased the expression of cytokines, chemokines, and cell migration-related genes. The promoter regions of the latter showed an enrichment of putative motifs for binding the transcription factors forkhead box O1 (FOXO1), FOXO3, NF-κB (NF-κB1), and RELA proto-oncogene and NF-κB subunit. Of note, FOXO1 inhibition by the FOXO1-selective inhibitor AS1842856 significantly reduced both migration and the expression of migration-related genes. In summary, our results indicate that TLR3 stimulation induces hMSC migration through the expression of FOXO1-activated genes.

Gastric cancer is a serious threat to human health, and its pathogenesis may be regulated by a variety of mRNAs. Abnormal expression of microRNA-340 has been frequently reported in many malignant neoplasms, while the molecular mechanism of miR-340 has not been explored in gastric cancer. In this study, the mRNA level of miR-340 was determined by real-time PCR in GC cell lines. The miR-340 mimic was transiently transfected into GC cells by using Lipofectamine™ 2000 reagent. The BrdU-ELISA results showed that introduction of miR-340 inhibited cell proliferation. It was demonstrated that miR-340 mimic arrested cell cycle progression and promoted apoptosis of MKN-45 and BGC-823 cells. In addition, the overexpression of miR-340 could inhibit invasion and EMT of MKN-45 and BGC-823 cells. The expression of NF-κB1 was evidently reduced by up-regulation of miR-340. Luciferase reporter assay further confirmed that miR-340 could directly target the 3'UTR of NF-κB1. Moreover, overexpression of NF-κB1 transfected with miR-340 mimic partially reversed the inhibitory of miR-340 mimic in MKN-45 and BGC-823 cells. In conclusion, miR-340 induced cell apoptosis and inhibited invasion by down-regulation of NF-κB1, which might be a potential target in treatment and prevention of gastric cancer.

The present study aimed to investigate the microRNA (miRNA) profile in human medullary thyroid carcinoma (MTC) tissue. The GSE40807 data profile was downloaded from the Gene Expression Omnibus database. Following preprocessing, differentially expressed microRNAs (DEMs) between MTC and healthy tissues were identified. Based on the obtained DEMs, transcription factor (TF)‑miRNA and miRNA‑target gene regulatory association pairs were predicted. Finally, functional enrichment analysis was performed on target genes of DEMs. Fifteen upregulated and 17 downregulated DEMs were identified. In the constructed TF‑miRNA regulatory network, hsa‑miR‑9‑5p was regulated by 9 TFs and hsa‑miR‑1 was regulated by 8 TFs. TFs of nuclear factor of κ light polypeptide gene enhancer in B‑cells 1 (NF‑κB1) and v‑myc avian myelocytomatosis viral oncogene homolog (MYC) regulated 4 and 3 DEMs, respectively. In the miRNA‑target gene regulatory network, hsa‑miR‑1, hsa‑miR‑9‑5p, hsa‑miR‑96‑5p and hsa‑miR‑590‑5p were most upregulated. The target genes of these 4 miRNAs were primarily enriched in the mitogen activated protein kinase (MAPK) signaling pathway. Therefore, MAPK signaling pathway may serve important roles in MTC progression. In conclusion, the DEMs hsa‑miR‑1 and hsa‑miR‑9‑5p, and TFs of NF‑κB1 and MYC may be used as biomarkers for the diagnosis and treatment of MTC.

UNASSIGNED

We aimed to evaluate whether the polymorphism of poly(ADP-ribose) polymerase-1 (PARP-1) is involved as potential risk factor in the development of spinal cord injury (SCI) among Chinese individuals.

UNASSIGNED

Patients with a confirmed diagnosis of SCI (other than traumatic injury) and healthy individuals with no clinical symptoms of SCI were enrolled at Spinal Cord Injury Care Center, The Third People's Hospital of Dalian, China. Genetic polymorphisms were studied in plasma samples by polymerase chain reaction-restriction fragment length polymorphism assay.

UNASSIGNED

A total of 130 Chinese patients with SCI and 130 healthy Chinese individuals were included. We found that patients with the GG genotype (odds ratio [OR]: 4.09, 95% confidence interval [CI] 2.42-6.90, P<0.001) and carriers of the G allele (OR 3.96, 95% CI 2.33-6.74, P<0.0001) were at high risk of developing SCI. A del/ins polymorphism of the NF-κB1 gene (OR 3.32, 95% CI 1.96-5.61, P<0.001) was also found to be associated with SCI.

UNASSIGNED

Our study suggests that PARP-1 polymorphisms are involved in the development of SCI in Chinese individuals. Thus, PARP-1 polymorphisms can be considered as one of the potential risk factors for developing SCI.

We have shown that NADPH oxidase (NOX)5-S may mediate the acid-induced decrease in cell apoptosis. However, mechanisms of NOX5-S-dependent decrease in cell apoptosis are not fully understood. In this study, we found that silencer-of-death domain (SODD) was significantly increased in esophageal adenocarcinoma (EA) tissues, EA cell lines FLO and OE33, and a dysplastic cell line CP-B. Strong SODD immunostaining was significantly higher in low-grade dysplasia (66.7%), high-grade dysplasia (81.2%), and EA (71.2%) than in Barrett's mucosa (10.5%). Acid treatment significantly increased SODD protein and mRNA expression and promoter activity in FLO cells, an increase that was significantly decreased by the knockdown of NOX5-S and nuclear factor κB (NF-κB)1 p50 with their small interfering RNAs. Similarly, acid-induced increase of SODD mRNA was blocked by knockdown of NOX5-S and p50 in a BE cell line CP-A. Overexpression of NOX5-S significantly increased SODD protein expression in FLO cells. Moreover, overexpression of NOX5-S or p50 significantly increased the SODD promoter activity and decreased the caspase 9 activity or apoptosis. NOX5-S overexpression-induced increase in SODD promoter activity was significantly decreased by knockdown of p50. In addition, acid treatment significantly decreased the caspase 9 activity, a decrease that was significantly inhibited by knockdown of SODD. Furthermore, chromatin immunoprecipitation assay showed that NF-κB1 p50 bound to SODD genomic DNA containing a NF-κB-binding element GGGGACACCCT. This binding element was further confirmed by a gel mobility shift assay. We conclude that acid-induced increase in SODD expression and decrease in cell apoptosis may depend on the activation of NOX5-S and NF-κB1 p50 in FLO cells.

To study the effect of intradiscal injection of simvastatin on discogenic pain in rats and its possible mechanism, 30 adult female rats were used in this experiment. Twenty rats were randomly divided into sham operation group (Control group), intervertebral disk degeneration group (DDD group), intervertebral disk degeneration + hydrogel group (DDD + GEL group), and intervertebral disk degeneration + simvastatin group (DDD + SIM group). The mechanical pain threshold and cold sensation in rats were measured. The contents of NF-kappa B1, RelA, GAP43, SP, CGRP, TRPM 8, IL-1β, and TNF-α in the intervertebral disk (IVD), the corresponding contents of dorsal root ganglion (DRG) and plantar skin GAP43 and TRPM 8 were quantitatively detected by PCR. The corresponding IVDs were stained to detect their degeneration. There was no significant difference in the mechanical pain threshold between the groups at each time point. From the first day to the 8th week after surgery, the cold-sensing response of the DDD group was significantly higher than that of the Control group (P < 0.05). At 7 and 8 weeks postoperatively, the cold-sensing response of the DDD + SIM group was significantly lower than that of the DDD + GEL group (P < 0.05). The levels of NF-κB1, RelA, GAP43, SP, CGRP, TRPM8, IL-1β, and TNF-α in the IVD of DDD + SIM group were significantly lower than those in DDD group (P < 0.05). The content of GAP43 and TRPM8 in rat plantar skin decreased significantly and TRPM8 in DRG decreased significantly (P < 0.05).

Keywords: GAP43=growth-associated peptide 43; NF-κB – Nuclear factor kappa B; cold sensitivity; discogenic low back pain; simvastatin.

OBJECTIVE

To delineate and confirm the signaling processes involving mitogen-activated protein kinase (MAPK) and Toll-like receptor (TLR) in the regulation of lipopolysaccharide (LPS)-induced activity of macrophages by genistein at the protein and transcriptional levels.

METHODS

RAW264.7 macrophages were treated with genistein and then stimulated with LPS (100 ng/mL) for different time duration. We evaluated the induction of MAPK phosphorylation by Western blot analysis; RT2 Profiler(TM); PCR array was used to investigate the expressions of TLR pathway-related genes after different treatments.

RESULTS

LPS led to the phosphorylation of Raf, MEK1/2 and ERK1/2 at 15 minutes post-stimulation and it lasted till 30 minutes. Phosphorylation of p38 was also observed, but it was not as obvious as p-ERK1/2. Addition of genistein further increased the phosphorylation of ERK1/2, its downstream protein p90rsk and p38. Cells treated with LPS and LPS together with genistein demonstrated significant number of genes to be differentially regulated as compared with control cells. Genistein alone could up-regulate the gene CD80, MEKK1, c-fos, Rela, and Ticam2. LPS could up-regulate 20 genes including cytokines IFN-β, IL-10, IL-1α, IL-1β, IL-6, TNF-α, colony stimulating factor 2 (CSF-2) and CSF-3, chemokines CCL2 and CXCL10, transcription factor NF-κB1, IκB-α and cyclooxygenase 2 (COX-2). The presence of genistein led to a strong inhibition of the expressions of these genes and up-regulated the transcription factors IκB-β and c-Rel, a subunit of NF-κB.

CONCLUSIONS

Genistein strongly enhances the LPS-induced activities of MAPK transduction cascades and inhibits TLR pathway.

Metabolic disorders are closely associated with dietary habits and seem to be related to neuroinflammation and neurodegenerative disease in humans. Emblica officinalis (EOT) fruits not only have good nutritional value but also have excellent therapeutic potential. We used a tannins-enriched fraction of EOT fruit with the expectation of controlling diet-induced neuroinflammation and cognitive impairment in rats. A high-salt and cholesterol diet (HSCD) was used to induce neuroinflammation and cognitive impairment in rats. The diet of the rats was then supplemented with EOT (100 and 200 mg/kg b.w.) for 7 weeks. In order to evaluate the neuroprotective effects of EOT; in silico study, neurobehavioral tests, biochemical analyses, and immunohistochemical studies were performed. In silico study of p50 (NF-κB1) receptors with emblicanin (the main constituent of EOT) suggests that EOT has binds to NF-κB. EOT treatment reversed the HSCD-induced behavioral and memory disturbances in a step-down-type passive avoidance test. EOT treatment also inhibited HSCD-induced NF-κB upstream signaling, including the release of Th1, such as TNF-α, and downstream signaling Th2, such as IL-10, by flow cytometer. In addition, EOT treatment attentuated the HSCD-induced increase in the level of cognitive impairment markers, such as amyloid β. Furthermore, immunohistochemical results demonstrated that EOT modulated neuronal cell death by inhibiting the overexpression of NF-kB in brain. This study confirms that EOT may be a promising therapy in ameliorating the neurotoxicity of HSCD; however further studies are warranted to elucidate the exact mechanism of action of EOT.

Ultraviolet (UV) irradiation affects neuronal structures of the skin and accelerates skin aging. Cytokine cascades in keratinocytes after UV irradiation may result in a paracrine inhibitory effect on nerve cells. The purpose of the present study was to determine the direct effect of cytokines induced by UV radiation on nerve cells in terms of neuronal senescence. Our group performed a preliminary study to determine cytokines induced in UV-irradiated keratinocytes. Among 40 cytokines studied, granulocyte-macrophage colony-stimulating factor (GM-CSF) was increased 4-fold in inflammation antibody array. The GM-CSF was added to cultured human neuroblastoma cells. To evaluate the effect of cellular senescence, the authors performed real-time polymerase chain reaction (RT-PCR), western blot, immunocytochemical, and phase-contrast microscopic evaluations. Expression levels of matrix metallopeptidase-9 (MMP-9), nuclear factor kappa-light-chain-enhancer of activated B cells 1 (NF-κB1), inducible nitric oxide synthase (iNOS), and interleukin β1 (IL-β1) were assessed by RT-PCR. Expression levels of AAP and beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) related to formation of beta-amyloid were evaluated by western blot analysis. Expression levels of MMP-9, NF-κB1, iNOS, and IL-β1 after treatment with GM-CSF were significantly higher than those in the control group. Enhanced expression of AAP and BACE1 was also observed in the treatment group. Thus, GM-CSF might have a provocative effect on nerve cells in terms of neuronal senescence.

B-cell lymphoma (Bcl)-3 is a nonclassical member of the IκB protein family known to interact with transcriptionally inactive NF-κB1 and NF-κB2 homodimers to modulate gene expression. Besides its action as an oncoprotein, Bcl-3 has been shown to have both proinflammatory and anti-inflammatory functions depending on the cell-type affected. In this issue of the European Journal of Immunology, Tassi et al. [Eur. J. Immunol. 2015. 45: 1059-1068] report that Bcl-3 inhibits the production of the proinflammatory chemokines CXCL9 and CXCL10 in keratinocytes, thereby restricting the influx of CD8(+) effector T cells in a mouse model of allergic contact dermatitis. In addition, mice with a global deficiency of Bcl-3 show enhanced ear swelling responses in the late phase of contact hypersensitivity responses. Besides keratinocytes, other radioresistant cell types appear to also utilize Bcl-3 to dampen the inflammatory response. This Commentary will discuss the evidence supporting Bcl-3 as a critical player in limiting inflammation during the later stages of contact hypersensitivity.

Exposure to tobacco smoke is a major public health concern that can also affect ophthalmic health. Based on previous work demonstrating the important role of the sympathetic nervous system (SNS) in corneal wound repair, we postulated that acute tobacco smoke exposure (ATSE) may act through the SNS in the impairment of corneal wound repair. Here we find that ATSE rapidly increases the markers of inflammatory response in normal corneal limbi. After an abrasion injury, ATSE exaggerates inflammation, impairs wound repair, and enhances the expression of nuclear factor-κB (NF-κB) and inflammatory molecules such as interleukin-6 (IL-6) and IL-17. We find that chemical SNS sympathectomy, local adrenergic receptor antagonism, NF-κB1 inactivation, and IL-6/IL-17A neutralization can all independently attenuate ATSE-induced excessive inflammatory responses and alleviate their impairment of the healing process. These findings highlight that the SNS may represent a major molecular sensor and mediator of ATSE-induced inflammation.

Streptocaulon juventas is a well-known plant that has antimicrobial activity, in vitro antiplasmodial activity, anti-proliferative activity, and antioxidant activity. In this study, we showed experimental evidence that proved that S. juventas root ethanolic extract has wound healing activities. First, in a mouse excision wound model, S. juventas root ethanolic extract at a dose of 100 mg/kg/day significantly reduced the wound closure time. After 7 days, the wound granulation tissue in mice treated with the extract exhibited a 2.3-fold decrease in inflammatory cells, a 1.7-fold increase in fibroblasts and enhanced angiogenesis. Molecular analysis also revealed that after wounds were treated with S. juventas root ethanolic extract, TNF-α and NF-κB1 gene expression were down-regulated by 4.7 and 3.7 times, respectively. In contrast, TGF-β1 and VEGF gene expression were up-regulated by 1.9 and 6.5 times, respectively. Taken together, our experimental data strongly show that the ethanolic extract from S. juventas root displays remarkable wound healing activity.

UNASSIGNED

The primary objective of this study was to evaluate whether the polymorphism of poly (ADP-ribose) polymerase-1 (PARP-1) is involved as a potential risk factor in the development of acute renal injury in elderly Chinese patients with diabetes mellitus.

UNASSIGNED

In this pilot study, diabetic patients of either gender (aged ≥65 years) with a confirmed diagnosis of acute renal injury and individuals with no clinical symptoms of acute renal injury were enrolled at Nanxishan Hospital of Guangxi Zhuang Autonomous Region, China. Genetic polymorphism of PARP-1 was assessed using polymerase chain reaction-restriction fragment length polymorphism assay. Cytokine levels (interleukin [IL]-6, IL-1β and tumor necrosis factor-alpha) were measured in the serum samples by sandwich enzyme-linked immunosorbent assay technique.

UNASSIGNED

A total of 130 Chinese patients with acute renal injury and 130 Chinese individuals with no clinical symptoms of acute renal injury were included. We found that the patients with GG genotype and carriers of the G and C alleles of PARP-1 were at high risk of developing acute renal injury. Moreover, del/ins polymorphism of the NF-κB1 gene was also found to be associated with acute renal injury. In addition, the levels of IL-6, IL-1β and tumor necrosis factor-alpha were significantly increased in patients with acute renal injury (p<0.05).

UNASSIGNED

Our findings showed the involvement of PARP-1 polymorphisms in the development of acute renal injury in Chinese individuals. This study identified the involvement of two SNPs of PARP-1 (C410T and G1672A) in development of acute renal injury among Chinese diabetic patients. Also, increased expression of C and G alleles of PARP-1 can be considered as one of the potential risk factors for developing acute renal injury. Increased serum cytokine levels can be considered as one of the potential risk factors for developing acute renal injury.

Type 2 diabetes mellitus (T2DM) accompanied by hyperlipidemia confers higher risk for diabetes as well as cardiovascular diseases. NF-κB is actively involved in generating low-grade inflammation and oxidative stress triggering the development of diabetic complications. In this study, we have attempted to investigate the association between NF-κB1 functional promoter polymorphism-94 ATTG insertion/deletion (rs28362491) with inflammatory markers in developing diabetes-linked dyslipidemia. We performed a case-control study in a total of 401 individuals belonging to three categories such as Type 2 diabetes with dyslipidemia, Type 2 diabetes without dyslipidemia, and normal healthy controls. Experiments were carried out using genotyping, real-time PCR, and western blot. Pearson's correlation, analysis of variance, and logistic regression were utilized for statistical analysis. As per genetic association conducted in this study the SNP rs28362491 showed significant allelic and genotypic associations (Allelic: OR = 1.374, CI 0.9797-1.927, p = 0.003, and Genotypic in dominant model: OR = 1.77, CI 1.04-2.99, p = 0.002) with the risk of diabetes and associated dyslipidemia. The -94 ATTG insertion/insertion (ins/ins) genotype was associated with significantly increased level of serum TNF-α (p = 0.002), serum IL-6 (p = 0.067) in diabetes-induced dyslipidemia. Multiple linear regression analysis identifies independent correlation of Total cholesterol, HDL, LDL, TNF-α, and rs28362491 ATTG ins/ins with triglyceride in diabetic dyslipidemic condition. T2DM with dyslipidemia having ins/ins genotype showed significant increased expression of pro-inflammatory cytokines such as TNF-α, IL-6, and activation of NF-κB. Our study reports that individuals with ATTG insertion allele and ATTG ins/ins genotype at NF-κB1 promoter regulatory gene predicts the risk and severity of T2DM-linked dyslipidemia.

Objective: To investigate the regulatory effect of the transcription factor NF-kB1 on the expression of miR-195 in prostate cancer (PCa).

Methods: We analyzed the possibility of NF-kB1 binding to the miR-195 promoter and the expression of NF-kB1 in PCa using the JASPAR and Oncomine databases, respectively, and determined the expressions of NF-kB1 and miR-195 in PCa cells by real-time quantitative PCR after inhibiting the former by interfering RNA targeting NF-kB1. We detected the activity of the luciferase reporter gene after constructing its gene plasmid in the miR-195 promoter region and having it co-transfected with the NF-kB1 plasmid. Then we analyzed the correlation between the expressions of miR-195 and NF-kB1 in the prostate tissue.

Results: NF-kB1 was overexpressed in PCa. After inhibition of the expression of NF-kB1, that of miR-195 was increased in PC-3 and DU-145 cell lines, with a negative correlation between the NF-kB1 and miR-195 expressions in the PCa tissue. The results of luciferase reporter gene assay showed direct binding of NF-kB1 to the miR-195 promoter zone.

Conclusions: NF-kB1 regulates the expression of miR-195 in prostate cancer.

Keywords: miR-195; prostate cancer; NF-kB1.

Background: Initiation, promotion, progression, and metastasis of mammary tumors are mediated by dysregulation of multiple genes involved in various signaling pathways. Expressional variation of these molecules significantly influences cancer cell proliferation in breast cancer.

Aims and objectives: In the current study, tumor necrosis factor-alpha (TNF-α) and its downstream effector nuclear factor kappa-B1 (NF-κB1) mean transcript levels were explored and associated with molecular subtypes in breast cancer cohort of Pakistan. Freshly excised tumors (n = 150) along with background tissues were collected for RNA isolation and cDNA synthesis.

Materials and methods: Quantitative polymerase chain reaction was carried out for quantification of TNF-α, NF-κB1, and β-actin gene transcripts along with estrogen receptor, progesterone receptor, HER2, and Ki-67, followed by statistical analysis.

Results: For TNF-α and NF-κB1, 95% and 77% of the cohort was found to be positive, respectively. Both of these molecules were found to be significantly upregulated in tumors when compared against their respective controls (P < 0.0001). Association of TNF-α and NF-κB1 with late clinical stages, poorly differentiated tumors, increased tumor size, nodal involvement, and metastasis was observed to be statistically significant (P < 0.05). Strong positive correlation was established between TNF-α and NF-κB1(r = 0.465, P< 0.05). Moreover, mean transcript levels of TNF-α and NF-κB1 were significantly elevated in Luminal A and Luminal B subtypes of breast cancer patients, respectively.

Conclusion: Strong positive correlation between TNF-α and NF-κB1 proposed the putative role of these molecules as prognostic biomarkers in breast cancer.

Keywords: Breast cancer; nuclear factor kappa B1; prognosis; quantitative polymerase chain reaction; tumor necrosis factor-α.

Background: Use of donation after circulatory death (DCD) hearts is becoming more prevalent in cardiac transplantation. However, there is no standardized approach to myocardial preservation, and little data exists on ultrastructural changes in DCD hearts. We have previously identified increased proapoptotic and proinflammatory activity in brain dead donor (BDD) hearts that subsequently exhibit primary graft failure and lower levels in DCD left atrial tissue. This study further investigates these markers and correlates them with cardiac function in DCD hearts.

Methods: This prospective study used donor hearts deemed unsuitable for transplant after gaining institutional ethics approval; 11 human hearts were obtained from 5 DCD donors and 6 BDDs. All hearts were preserved by continuous microperfusion for 4 hours with a cold crystalloid solution and then were evaluated on a blood perfusion bench rig. After 4 hours perfusion and working assessment, tissues from all cardiac chambers were stored for later messenger RNA (mRNA) analysis for proapoptotic and proinflammatory markers.

Results: Significantly raised levels of caspase-1, BNIP3, and NADPH oxidase mRNA expression were identified in cardiac chambers from BDD hearts compared to DCD hearts, and these differences were exaggerated in older donors. In the pooled analysis, lower expression of caspase-1, NF-κB1, and BNIP3 mRNA correlated with developed pressure at 1 hour after reperfusion in the right ventricle, but not the left.

Conclusion: Compared to BDD hearts, DCD hearts exhibit less stimulation of proapoptotic cascades and reactive oxygen species, potentially reducing their susceptibility to ischemic reperfusion injury.

Drug addiction is a chronic relapsing brain disease. Alterations of glucose uptake and metabolism are found in the brain of drug addicts. Insulin mediates brain glucose metabolism and its abnormality could induce brain injury and cognitive impairment. Here, we established a rat model of phenobarbital addiction by 90 days of dose escalation and evaluated addiction-related symptoms. We also performed ¹⁸F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) to detect glucose uptake in the brain and proteomic analysis of the function of the differentially expressed (DE) proteins via bioinformatics in brain tissues by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) on days 60 and 90 of phenobarbital or 0.5% carboxymethyl cellulose sodium (CMC-Na) (vehicle) administration. The results showed that phenobarbital-addictive rats developed severe withdrawal symptoms after abstinence and glucose uptake was significantly increased in the brain. Proteomics analysis showed that numerous DE proteins were enriched after phenobarbital administration, among which CALM1, ARAF, and Cbl proteins (related to the insulin signaling pathway) were significantly downregulated on day 60 but not day 90. However, SLC27A3 and NF-κB1 proteins (related to insulin resistance) were significantly upregulated on day 90 (data are available via ProteomeXchange with identifier PXD021101). Our data indicate that the insulin signaling pathway and insulin resistance may play a role in the development of phenobarbital addiction and brain injury, so the findings may have important clinical implications.

Keywords: PET-CT; addiction; glucose uptake; insulin resistance; insulin signaling pathway; phenobarbital; proteomics.