Ribonucleic Acid Levels Research Articles

Seed priming with melatonin: A promising approach to combat abiotic stress in plants

Melatonin is an indolic compound derived from tryptophan and has an important role in the general processes of plants including germination, photosynthesis, circadian rhythms, flowering, senescence, and alleviating stress. Integrating melatonin application to agronomical practices reportedly provides numerous benefits. More particularly seed priming with melatonin has been shown to improve seed germination as well as improve resilience under adverse conditions. This review discusses in detail the studies showing elevated drought, salinity, heat, and heavy metal stress resistance as a result of melatonin seed priming. Differences in transcript and protein levels have been observed in plants/seedlings germinated via melatonin primed seeds, providing the foundation for further studies. Many pieces of research have also shown primed seeds possess a greater content of osmoprotectants, upregulation of energy-producing pathways, and an increase in protein content, which may lead to enhancing abiotic stress response in plants. The primary function attributed to melatonin is reactive oxygen species scavenging, which consequently leads to enhancing plant stress resistance against multiple stresses. Here, we highlight all the potential mechanisms involved in melatonin meditated induction of abiotic stress tolerance. We review the reports emphasizing the role played by melatonin under stress conditions in multiple crop species, the modification at genome, ribonucleic acid, and protein level involved, and the underlying mechanisms. Information provided here will be helpful to better understand the melatonin role in plants and its efficient exploration in the cropping system.

Discoidin domain receptor 1 as a promising biomarker for high-grade gliomas.

Two fundamental challenges in the current therapeutic approach for central nervous system tumors are the tumor heterogeneity and the absence of specific treatments and biomarkers that selectively target the tumor tissue. Therefore, we aimed to investigate the potential relationship between discoidin domain receptor 1 (DDR1) expression and the prognosis and characteristics of glioma patients. Tissue and serum samples from 34 brain tumor patients were evaluated for DDR1 messenger ribonucleic acid levels in comparison to 10 samples from the control group, and Kaplan-Meier survival analysis has performed. DDR1 expression was observed in both tissue and serum samples of the patient and control groups. DDR1 expression levels in tissue and serum samples from patients were higher in comparison to the control group, although not statistically significant (P > 0.05). A significant correlation between tumor size and DDR1 serum measurements at the level of 0.370 was reported (r = 0.370; P = 0.034). The levels of DDR1 in serum showed a positive correlation with the increasing size of tumor. The results of the 5-year survival analysis depending on the DDR1 tissue levels showed a significantly higher survival rate (P = 0.041) for patients who have DDR1 tissue levels above cutoff value. DDR1 expression was significantly higher among brain tumor tissues and serum samples and its levels showed a positive correlation with the increased size of tumor. This study can be a starting point, since it investigated and indicated, for the first time, that DDR1 can be a novel therapeutic and prognostic target for aggressive high-grade gliomas.

Effect of omentin-1 on cancer stem cell surface markers and tumour-suppressive miRNA expression in a high-glucose environment associated with colorectal cancer.

To investigate how omentin-1 impacts colorectal cancer stem cell surface markers and the expression levels of tumour-suppressive micro ribonucleic acid in a colorectal cancer-associated high-glucose environment. The study was conducted in the First Affiliated Hospital of Anhui Medical University，Anhui, China，from April 2018 to January 2019 and comprised cluster of differentiation133 and colorectal cancer stem cells from the SW480 cell line（the human colon adenocarcinoma cell line） obtained through immunomagnetic beads-based cell isolation. The colon cancer stem cells were divided into 6 groups: Z0 (control), Z1 (1ug/mL omentin-1), Z2 (2ug/mL omentin-1), G0 (5.0g/mL glucose), G1 (1ug/mL omentin-1 and 5.0g/mL glucose), and G2 (2ug/mL omentin-1 and 5.0 g/mL glucose). After 24 hours of intervention, quantitative polymerase chain reaction and western blot test were used for the detection of messenger ribonucleic acid and protein levels of stem cell surface markers. The colorectal cancer stem cells were divided into three groups: the control group, omentin group 1 (1ug/mL omentin-1) and omentin group 2 (2ug/mL omentin-1). After 24 hours of intervention, the expression of tumour suppressor micro ribonucleic acid was measured using quantitative polymerase chain reaction. Data was analysed using SPSS 23. Compared to the Z0 group, cluster of differentiation133 messenger ribonucleic acid expression reduced sharply in Z1 group (p<0.05), while Z2 group saw a marked increase in the expression (p<0.05). With respect to tumour-suppressive micro ribonucleic acid expression, micro ribonucleic acid 126, 145, 34a and 342-5P in omentin group 2 exhibited an expression level significantly higher than those in the control group and the omentin group 1 (p<0.05). High glucose levels were found to upregulate the expression of colorectal cancer stem cell surface markers cluster of differentiation133 messenger ribonucleic acid and protein. Also, omentin-1 was found to be associated with the downregulation of cluster of differentiation133 messenger ribonucleic acid and protein expression and the upregulation of cluster of differentiation 44 messenger ribonucleic acid expression in a high-glucose environment. Finally, omentin-1 was found to have the ability to promote the expression of relevant tumour-suppressive micro ribonucleic acids 126, 14, 34a and 342-5P.

Methyltransferase-like 3 upregulation is involved in the chemoresistance of non-small cell lung cancer.

BackgroundTreatments for non-small cell lung cancer (NSCLC) have improved tremendously, but therapeutic resistance is a common and major clinical challenge in treatment. Methyltransferase-like 3 (METTL3) is a ribonucleic acid (RNA) methyltransferase that has crucial functions in the development and progression of cancers, including drug resistance, by regulating N6-methyladenosine (m6A) modification. However, the role of METTL3 in the progression and drug resistance of NSCLC is poorly understood.MethodsThe expression levels of METTL3 and AKT serine/threonine kinase 1 (AKT1) in NSCLC tissues were detected using quantitative real-time PCR (RT-qPCR), Western blots, and immunohistochemical assays. The m6A levels of AKT1 messenger RNA (mRNA) in NSCLC tissues were detected using m6A methylated RNA immunoprecipitation–quantitative polymerase chain reaction.ResultsThe expression levels of METTL3 and the AKT1 protein were significantly increased in NSCLC tissues, and m6A expression levels of AKT1 mRNA were dramatically upregulated in NSCLC tissues. Additionally, METTL3, AKT1 protein, and m6A levels of AKT1 mRNA were overexpressed in chemoresistant NSCLC samples, and high expression levels of METTL3 and AKT1 were correlated with poor patient survival, especially in chemoresistant NSCLC patients. Further, AKT1 protein expression and m6A levels of AKT1 mRNA were positively correlated with METTL3 expression, and AKT1 protein expression was positively correlated with m6A levels of AKT1 mRNA. Moreover, METTL3 and AKT1 protein expression levels were significantly associated with cisplatin susceptibility, tumor, node, metastasis stage, and lymph node metastasis.ConclusionsTaken together, our results indicate that METTL3 contributes to the progression and chemoresistance of NSCLC by promoting AKT1 protein expression through regulating AKT1 mRNA m6A levels, and may provide an efficient therapeutic intervention target for overcoming chemoresistance in NSCLC.

Effect of Prostaglandin E2 Agonist Omidenepag on the Expression of Matrix Metalloproteinase in Trabecular Meshwork Cells

PurposeTo investigate the effects of prostaglandin E2 agonist omidenepag (OMD) on the expression of matrix metalloproteinase (MMP) in human trabecular meshwork (TM) cells.MethodsPrimarily cultured human TM cells were exposed to 0, 1, 10, or 40 μmol/L OMD for 3 days. The permeability through the TM cell monolayer was assessed using carboxyfluorescein. Expressions of messenger ribonucleic acid and protein levels of MMP-1, MMP-3, and MMP-9 were measured by reverse transcription polymerase chain reaction and Western blotting, respectively. Also, the permeability, expression of messenger ribonucleic acid, and protein levels of MMPs were measured after exposure to 1 μmol/L latanoprost free acid (LAT).ResultsOMD and LAT did not affect the cellular survival (all p > 0.05). Each concentration of OMD and LAT did not affect the permeability of carboxyfluorescein significantly (all p > 0.05). LAT increased the level of MMP-1 protein but did not increase the levels of MMP-3 and MMP-9 proteins. Each concentration of OMD did not affect the levels of MMP-1, MMP-3, and MMP-9 proteins (all p > 0.05)ConclusionsIn TM cells, prostaglandin E2 agonist OMD did not increase the permeability through the TM cell monolayer, and the protein levels of MMPs. These suggest that the direct effect on the trabecular outflow by OMD may be limited.

Temporal characteristics of astrocytic activation in the TNC in a mice model of pain induced by recurrent dural infusion of inflammatory soup

BackgroundAstrocytic activation might play a significant role in the central sensitization of chronic migraine (CM). However, the temporal characteristics of the astrocytic activation in the trigeminal nucleus caudalis (TNC) and the molecular mechanism under the process remain not fully understood. Therefore, this study aims to investigate the duration and levels change of astrocytic activation and to explore the correlation between astrocytic activation and the levels change of cytokines release.MethodsWe used a mice model induced by recurrent dural infusion of inflammatory soup (IS). The variation with time of IS-induced mechanical thresholds in the periorbital and hind paw plantar regions were evaluated using the von Frey filaments test. We detected the expression profile of glial fibrillary acidic protein (GFAP) in the TNC through immunofluorescence staining and western blot assay. We also investigated the variation with time of the transcriptional levels of GFAP and ionized calcium binding adapter molecule 1 (Iba1) through RNAscope in situ hybridization analysis. Then, we detected the variation with time of cytokines levels in the TNC tissue extraction and serum, including c-c motif chemokine ligand 2 (CCL2), c-c motif chemokine ligand 5 (CCL5), c-c motif chemokine ligand 7 (CCL7), c-c motif chemokine ligand 12 (CCL12), c-x-c motif chemokine ligand 1 (CXCL1), c-x-c motif chemokine ligand 13 (CXCL13), interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), macrophage colony-stimulating factor (M-CSF), interleukin 1beta (IL-1β), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 17A (IL-17A).ResultsRecurrent IS infusion resulted in cutaneous allodynia in both the periorbital region and hind paw plantar, ranging from 5 d (after the second IS infusion) to 47 d (28 d after the last infusion) and 5 d to 26 d (7 d after the last infusion), respectively. The protein levels of GFAP and messenger ribonucleic acid (mRNA) levels of GFAP and Iba1 significantly increased and sustained from 20 d to 47 d (1 d to 28 d after the last infusion), which was associated with the temporal characteristics of astrocytic activation in the TNC. The CCL7 levels in the TNC decreased from 20 d to 47 d. But the CCL7 levels in serum only decreased on 20 d (1 d after the last infusion). The CCL12 levels in the TNC decreased on 22 d (3 d after the last infusion) and 33 d (14 d after the last infusion). In serum, the CCL12 levels only decreased on 22 d. The IL-10 levels in the TNC increased on 20 d.ConclusionsOur results indicate that the astrocytic activation generated and sustained in the IS-induced mice model from 1 d to 28 d after the last infusion and may contribute to the pathology through modulating CCL7, CCL12, and IL-10 release.

Outreach onsite treatment with a simplified pangenotypic direct-acting anti-viral regimen for hepatitis C virus micro-elimination in a prison.

BACKGROUNDPrisoners are at risk of hepatitis C virus (HCV) infection, especially among the people who inject drugs (PWID). We implemented an outreach strategy in combination with universal mass screening and immediate onsite treatment with a simplified pan-genotypic direct-acting antivirals (DAA) regimen, 12 wk of sofosbuvir/velpatasvir, in a PWID-dominant prison in Taiwan.AIMTo implement an outreach strategy in combination with universal mass screening and immediate onsite treatment with a simplified pan-genotypic DAA regimen in a PWID-dominant prison in Taiwan.METHODSHCV-viremic patients were recruited for onsite treatment program for HCV micro-elimination with a pangenotypic DAA regimen, 12 wk of sofosbuvir/ velpatasvir, from two cohorts in Penghu Prison, either identified by mass screen or in outpatient clinics, in September 2019. Another group of HCV-viremic patients identified sporadically in outpatient clinics before mass screening were enrolled as a control group. The primary endpoint was sustained virological response (SVR12, defined as undetectable HCV ribonucleic acid (RNA) 12 wk after end-of-treatment).RESULTSA total of 212 HCV-viremic subjects were recruited for HCV micro-elimination campaign; 91 patients treated with sofosbuvir/Ledipasvir or glecaprevir/ pibrentasvir before mass screening were enrolled as a control. The HCV micro-elimination group had significantly lower proportion of diabetes, hypertension, hyperlipidemia, advanced fibrosis and chronic kidney diseases, but higher levels of HCV RNA. The SVR12 rate was comparable between the HCV micro-elimination and control groups, 95.8% (203/212) vs 94.5% (86/91), respectively, in intent-to-treat analysis, and 100% (203/203) vs 98.9% (86/87), respectively, in per-protocol analysis. There was no virological failure, treatment discontinuation, and serious adverse event among sofosbuvir/velpatasvir-treated patients in the HCV micro-elimination group.CONCLUSIONOutreach mass screening followed by immediate onsite treatment with a simplified pangenotypic DAA regimen, sofosbuvir/velpatasvir, provides successful strategies toward HCV micro-elimination among prisoners.

Inhibitory Effect of Polyphyllin D on Proliferation of Human Colon Cancer Cell Line SW480 via the MicroRNA-21/Phosphatase and Tensin Homolog Deleted on Chromosome 10

To evaluate the effects of polyphyllin D on the proliferation and apoptosis of human colon cancer SW480 cell lines and to reveal whether this process is mediated by the micro ribonucleic acid-21/phosphatase and tensin homolog deleted on chromosome 10 pathway. Human colon cancer SW480 cells were transfected with micro ribonucleic acid-21-mimic or mimic-negative control and treated with polyphyllin D at different concentrations (control, 0.25, 0.5, 1 and 2 μM) for 24 h or 48 h. Compared with control polyphyllin D group, the viability of human colon cancer SW480 cells significantly declined but the apoptosis rate significantly rose in 0.5 μM polyphyllin D group and the colony number of SW480 cells significantly decreased in 0.25 μM polyphyllin D group (p<0.05). Compared with control group, 2 μM polyphyllin D group had significantly increased protein expressions of Bcl-2 associated X protein and cleaved caspase-3, but decreased protein expression of B-cell lymphoma-2 in SW480 cells (p<0.001). The protein levels of micro ribonucleic acid-21 and phosphorylated-protein kinase B in SW480 cells were significantly lower, while the phosphatase and tensin homolog deleted on chromosome 10 messenger ribonucleic acid and protein levels were higher in 2 μM polyphyllin D group than those in control group (p<0.001). However, the protein expression of protein kinase B was not changed in 2 μM polyphyllin D group (p>0.05). After transfection with micro ribonucleic acid-21-mimic, the level of micro ribonucleic acid-21 in SW480 cells was significantly higher, while the phosphatase and tensin homolog deleted on chromosome 10 messenger ribonucleic acid and protein levels were significantly lower than those in control group (p<0.001). Compared with control group, the antiproliferative and pro-apoptotic effects of polyphyllin D on SW480 cells were reversed by transfection with micro ribonucleic acid-2-mimic (p<0.001). Polyphyllin D inhibits proliferation and induces apoptosis of human colon cancer SW480 cells via the micro ribonucleic acid-21/phosphatase and tensin homolog deleted on chromosome 10 pathway.

Effects of 7,12-Dimethylbenz(a)anthracene on Apoptosis of Breast Cancer Cells through Regulating Expressions of Fas Ligand and B-Cell Lymphoma 2

To provide reference basis for the apoptosis of breast cancer cells and the carcinogenesis of breast cancer, the effects of 7,12-dimethylbenz(a)anthracene on apoptosis regulators Fas ligand and B-cell lymphoma-2 were investigated. In this study, 62 female C57BL/6 mice aged from 4 w to 6 w were randomly divided into control group and test group, with 31 mice in each group. The test group was given 7,12-dimethylbenz(a) anthracene solution by gavage at a dose of 50 mg/kg and the control group was given normal saline of equal volume. On the 2nd d after the experiment, all the mice were killed by cervical dislocation. The results showed that after hematoxylin-eosin staining, the tumor cells in the test group were stacked up to form a substantial structure. The expression level of Fas ligand protein in the control group was greatly lower than that in the test group and the positive rate was 20.25 %, which was greatly lower than that of 89.65 % in the test group (p<0.01). The expression level of B-cell lymphoma-2 protein in the mammary gland tissues of mice in the test group was greatly higher than that of the control group and its positive rate was 87.96 %, which was greatly higher than that of 31.48 % in the control group (p<0.01). The expression levels of Fas ligand messenger ribonucleic acid in the mammary gland tissues of mice in the test group and control group were 5.82±4.37 and 1.27±0.12, respectively and there was a statistically obvious difference (p<0.05). The messenger ribonucleic acid expression levels of B-cell lymphoma-2 in the test group and the control group were 18.97±2.65 and 2.02±0.54, respectively, and there was an extremely obvious difference (p<0.01). The apoptosis rate of mammary gland cells in the test group was (19.79±3.53) % and that in the control group was (2.93±0.28) % and there was an extremely obvious difference (p<0.01). It indicated that 7,12-dimethylbenz(a)anthracene inhibited the apoptosis of breast cancer cells by regulating the upregulation of Fas ligand and B-cell lymphoma-2 expression.

Risk Factors for Pulmonary Metastasis in Differentiated Thyroid Carcinoma Patients and the Significance of Changes in Matrix Metalloproteinase 13 and microRNA-142 Levels.

This work aims to explore the risk factors of lung metastasis (LM) in differentiated thyroid cancer (DTC) (LM-DTC) and the effect of treatment and to detect the relationship between LM-DTC and the levels of matrix metalloproteinase-13 (MMP-13) and micro ribonucleic acid (RNA)-142 (miR-142) in peripheral blood. The data of 420 patients with DTC who are admitted from March 2020 to December 2021 are collected and divided into a non-metastasis group (non-LM group) of 400 cases and metastasis group (LM group) of 20 cases according whether the mung metastasis is found. In addition, risk factors of LM-DTC are analysed and compared. The results of multivariate logistic analysis show that age, disease course, and imaging timing are independent influencing factors of the radionuclide treatment effect. Follicular carcinoma, abnormal expressions of MMP-13, and miR-142 can increase the risk of LM-DTC. MMP-13 and miR-142 can be undertaken as auxiliary diagnostic biological indicators.

Effect of Dieda Ointment on Bone Morphogenetic Protein-2/Suppressor of Mothers Against Decapentaplegic 1/Runt-Related Transcription Factor 2/Sp7 Transcription Factor Pathway for Fracture Healing in Rats

Dieda ointment, a Chinese medicine, has been widely used in the clinical treatment of fracture after years of clinical research to prove that it has significant effects on healing injury and relieving pain, reinforcing tendons and setting bones. However, the mechanism of its effects on fracture healing is still unclear. The purpose of this study was to explore the mechanism of Dieda ointment in promoting fracture healing. The materials and methods explored were used to establish a fracture model of Sprague-Dawley rats, which was externally applied with Dieda ointment for 1, 2 and 4 w and imaging examination of the fracture site was performed. Histological examination was also performed. Western blotting and quantitative reverse transcription polymerase chain reaction were used to detect the expression of proteins and its messenger ribonucleic acid. We found that Dieda ointment can effectively accelerate callus formation at tibia fractures in rats and increase the messenger ribonucleic acid and protein expression levels of bone morphogenetic protein-2, suppressor of mothers against decapentaplegic 1, runt-related transcription factor 2 and Sp7 transcription factor, regulate bone formation and promote fracture healing. The study on the mechanism of Dieda ointment promoting fracture healing, not only provides theoretical and experimental basis for the subsequent study of Dieda ointment.

Interference with Tripartite Motif-Containing Protein 26 Expression on Proliferation of Glioma Cells

This research aims to study the effect of interference with tripartite motif-containing protein 26 expression on the proliferation of intracranial glioma cells. This research randomly selected 60 cases of intracranial glioma specimens and the brain tissue of 10 cases of brain trauma patients as a normal control group. First, cell culture is performed to observe the growth status of the cells. Subsequently, the small interfering ribonucleic acid interference technique was used to interfere with the expression of tripartite motif-containing protein 26 on the glioma cell lines A172 and U251 cells. The cells were collected for tripartite motif-containing protein 26, Western blot and fluorescence quantitative polymerase chain reaction was used to verify the knockout effect, and the cell proliferation, clone formation, transfer, and invasion experiments were performed. Experimental data showed that among 60 glioma patients, tripartite motif-containing protein 26 samples were positive in 46 cases and 10 normal brain tissue samples were all negative. The performance of tripartite motif-containing protein 26 in glioma was compared with the performance in normal brain tissue. The difference was statistically significant (p<0.01). The results show that tripartite motif-containing protein 26 can reduce the protein level of pre-B-cell leukemia homeobox through the correlation of time gradient and concentration gradient. Tripartite motif-containing protein 26 can reduce the messenger ribonucleic acid level of ring finger protein 6 and the downstream target gene of pre-B-cell leukemia homeobox without affecting the messenger ribonucleic acid level of pre-B-cell leukemia homeobox.

Study on the Mechanism of Zhenwu Decoction in Decreasing the Fats in Non-Alcoholic Fatty Liver Disease Using System Pharmacology and Cytological Validation Test

Purpose of this study was to screen the active compounds of Zhenwu decoction and their gene targets in the treatment of non-alcoholic fatty liver disease by network pharmacology and cytological validation test. The candidate compounds and related targets of Zhenwu decoction were obtained from traditional Chinese medicine system pharmacology database and PharmMapper. The non-alcoholic fatty liver disease-related genes were obtained from online Mendelian inheritance in man, GeneCards and DisGeNET databases. Molecular docking was carried out to simulate the binding affinities between potential core compounds and key target genes, and the chemical constituents of Zhenwu decoction were analyzed and identified by Ultra-Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry technology. Therapeutic effect and mechanism of Zhenwu decoction on non-alcoholic fatty liver disease were validated using in vitro analysis. The protein-protein interaction network analysis identified the albumin, protein kinase B1, epidermal growth factor receptor, caspase 3 and peroxisome proliferator activated receptor gamma as the key target genes. Gene ontology annotation analysis showed that the Zhenwu decoctionnon- alcoholic fatty liver disease target genes were mainly involved in the response to steroid hormone and lipid catabolic process. The results of the Kyoto encyclopedia of genes and genomes pathway enrichment analysis were mainly related to the Forkhead box O signaling pathway and phosphatidylinositol 3-kinaseprotein kinase B signaling pathway. Paeoniflorgenone had the highest binding affinities for albumin, protein kinase B1, epidermal growth factor receptor, caspase 3 and peroxisome proliferator activated receptor gamma. The in vitro experiment showed that 19.5 mg/ml (p≤0.05) and 39 mg/ml (p≤0.01) of Zhenwu decoction could reduce lipid accumulation. Real-time quantitative polymerase chain reaction analyses revealed that Zhenwu decoction induced down-regulation of epidermal growth factor receptor messenger ribonucleic acid levels and up-regulation of heat shock protein 90 alpha family class a member 1, mitogen-activated protein kinase 1 and phosphatidylinositol 3-kinase messenger ribonucleic acid levels.

The Molecular Mechanism of Compatibility of Achyranthes bidentata and Radix Paeoniae Alba Mediating Antioxidant Damage

To explore the molecular mechanism of antioxidant damage of Achyranthes bidentata and Radix Paeoniae Alba is the main objective of study. Rats were treated with Achyranthes bidentata and Radix Paeoniae Alba. The total protein, nuclear protein and total ribonucleic acid were extracted. The protein levels and messenger ribonucleic acid levels were detected by western blot and quantitative polymerase chain reaction, respectively. The apoptosis of cells was detected and the cell viability and clone formation experiments were carried out. The serum malondialdehyde content of the Achyranthes bidentata and white peony group was lower than that of the control group, while the superoxide dismutase activity was higher than that of the control group, which was statistically significant (p<0.05). The level of nuclear factor erythroid 2-related factor 2 protein in the nucleus increased after treatment with medicated serum. The protein levels and the messenger ribonucleic acid levels of heme oxygenase 1 and superoxide dismutase increased significantly (p<0.05). The percentage of apoptosis, increased cell viability and number of clones treated with medicated serum were significantly higher than those treated with serum, indicating that the compatibility of Achyranthes bidentata and white peony can promote the survival of cells under oxidative stress. Under the joint action of protein kinase C delta and protein kinase, Achyranthes bidentata and Radix Paeoniae Alba up-regulate the expression of antioxidant enzymes.

Crucial Role of Central Nervous System as a Viral Anatomical Compartment for HIV-1 Infection.

The chronic infection established by the human immunodeficiency virus 1 (HIV-1) produces serious CD4+ T cell immunodeficiency despite the decrease in HIV-1 ribonucleic acid (RNA) levels and the raised life expectancy of people living with HIV-1 (PLWH) through treatment with combined antiretroviral therapies (cART). HIV-1 enters the central nervous system (CNS), where perivascular macrophages and microglia are infected. Serious neurodegenerative symptoms related to HIV-associated neurocognitive disorders (HAND) are produced by infection of the CNS. Despite advances in the treatment of this infection, HAND significantly contribute to morbidity and mortality globally. The pathogenesis and the role of inflammation in HAND are still incompletely understood. Principally, growing evidence shows that the CNS is an anatomical reservoir for viral infection and replication, and that its compartmentalization can trigger the evolution of neurological damage and thus make virus eradication more difficult. In this review, important concepts for understanding HAND and neuropathogenesis as well as the viral proteins involved in the CNS as an anatomical reservoir for HIV infection are discussed. In addition, an overview of the recent advancements towards therapeutic strategies for the treatment of HAND is presented. Further neurological research is needed to address neurodegenerative difficulties in people living with HIV, specifically regarding CNS viral reservoirs and their effects on eradication.

The upregulation of proteins light chain 3 and autophagy-related 5 and the occurence of intestinal-type gastric cancer.

This study aims to investigate the expression levels and values of autophagy genes light chain 3 (LC3) and autophagy-related 5 (ATG5) in intestinal-type gastric cancer. Ninety samples of normal gastric mucosa, intraepithelial neoplasia, and gastric cancer tissue were used in this study. The messenger ribonucleic acid (mRNA) and protein expression levels of autophagy genes LC3 and ATG5 were detected using quantitative reverse transcription polymerase chain reaction, Western blot, and the immunohistochemistry method. The correlations of the autophagy genes and certain clinical pathological parameters were analyzed. The results showed that LC3 mRNA expression was 43.76 ± 20.31 in the normal group, 111.29 ± 18.65 in the intraepithelial neoplasia group, and 131.78 ± 26.29 in the gastric cancer group, while ATG5 mRNA expression was 4.52 ± 2.37 in the normal group, 7.09 ± 1.88 in the intraepithelial neoplasia group, and 10.25 ± 2.81 in the gastric cancer group. The differences between the groups were statistically significant (P < 0.05). The protein expression of LC3 in the normal group was 1.05 ± 0.41, 1.53 ± 0.36 in the intraepithelial neoplasia group, and 1.99 ± 0.14 in the gastric cancer group. The protein expression of ATG5 was 0.78 ± 0.24 in the normal group, 1.37 ± 0.39 in the intraepithelial neoplasia group, and 2.04 ± 0.63 in the gastric cancer group. The differences between the groups were statistically significant (P < 0.05). The positive rate of LC3 protein expression was 33.3% in the normal group and 60% in the intraepithelial neoplasia group, and the difference was statistically significant (χ2 = 4.89; P = 0.04). In the gastric cancer group, the positive rate of LC3 protein expression was 83.3%, making it significantly higher than the intraepithelial neoplasia group, with a statistically significant difference (χ2 = 4.02, P = 0.045). The positive rate of ATG5 protein expression was 23.3% in the normal group, 50.0% in the intraepithelial neoplasia group, and 76.7% in the gastric cancer group. The expression in the intraepithelial neoplasia group was much higher than in the normal group, with a statistically significant difference (χ2 = 4.59, P = 0.03), and that of the gastric cancer group was much higher than that of the intraepithelial neoplasia group, with a statistically significant difference (χ2 = 4.59, P = 0.03). LC3 protein expression was significantly correlated with depth of infiltration, and lymph node status. ATG5 protein expression was significantly correlated with age, depth of infiltration, and lymph node status. There was also a correlation between the LC3 and ATG5 proteins (correlation coefficient r = 0.72, P = 0.001). The enhanced autophagy activity of LC3 and ATG5 may participate in the occurrence and development of intestinal gastric cancer, and they may play a synergistic role in promoting the occurrence and development of intestinal gastric cancer. These findings provide clinical value for the diagnosis of intestinal gastric cancer.

Ocular expression of cyclin-dependent kinase 5 in patients with proliferative diabetic retinopathy.

ABSTRACTAims/IntroductionInhibition of peroxisome proliferator‐activated receptor gamma (PPARγ) phosphorylation mediated by cyclin‐dependent kinase 5 (Cdk5) is one of the main mechanisms of action of antidiabetic drugs. In this study, we analyzed the ocular expression and activation of Cdk5 in patients with proliferative diabetic retinopathy (PDR).Materials and MethodsThe concentrations of PPARγ, Cdk5 and its activating subunit (p35) were determined in the vitreous body of 24 PDR and 63 control eyes by enzyme‐linked immunosorbent assay. In addition, the messenger ribonucleic acid and protein expression levels of PPARγ, Cdk5 and p35 were measured in proliferative neovascular membranes from seven PDR eyes and non‐neovascular epiretinal membranes from five control eyes by quantitative real‐time polymerase chain reaction and immunohistochemical analysis.ResultsPPARγ, Cdk5 and p35 concentrations in the vitreous body were significantly higher in the PDR group compared with the control group. There was also a positive significant correlation of Cdk5 with PPARγ and p35 in the PDR group. Furthermore, the messenger ribonucleic acid expression levels of PPARγ, Cdk5 and p35 in proliferative neovascular membranes were significantly higher in the PDR group compared with the control group. Immunostaining showed increased protein expression levels of PPARγ, Cdk5 and p35 in proliferative neovascular membranes in the PDR group compared with the control group.ConclusionsCdk5 activation is involved in PDR pathogenesis through PPARγ expression, and inhibition of Cdk5‐mediated PPARγ phosphorylation might be a new therapeutic target for treatment of PDR.

Effectiveness of Nicotinamide Phosphoribosyltransferase/Pre-B Cell Colony-enhancing Factor/Visfatin in preventing High Glucose-induced Neurotoxicity in an In-vitro Model of Diabetic Neuropathy.

Diabetic neuropathy is a well-known complication of diabetes. Recently, hyperglycemia-induced toxicity has been confirmed to participates in multiple cellular pathways typical for neural deterioration. Nicotinamide phosphoribosyltransferase/pre-b cell colony-enhancing factor (Nampt/PBEF)/visfatin is a novel endogenous ligand that some studies have shown its neuroprotective effects on neurodegenerative disease. Therefore, we hypothesized that visfatin may prevent high glucose (HG)-induced neurotoxicity by inhibiting apoptosis, autophagy, and reactive oxygen species (ROS) responses properly. In this study, pheochromocytoma cell line 12 (PC12) cells were exposed to both HG concentrations (50, 75, 100, 125, 150 mM) and visfatin (50, 100, 150 ng/mL) at different time -points to determine the optimum time and dose of glucose and visfatin. To investigate the effects of visfatin on HG-induced damage in the PC12 diabetic neuropathy model, we examined ROS response, apoptosis, and autophagy using ROS detection kit, flow cytometry, and real-time PCR/Western blot, respectively. We determined that HG concentration significantly increased the ROS level and apoptosis of diabetic PC12 cells. However, visfatin treatment significantly decreased the ROS production (P<0.05) and apoptosis of diabetic PC12 cells (P<0.0001). Beclin-1 messenger ribonucleic acid (mRNA) level (P<0.05) and light chain 3 (Lc3)-II protein level (P<0.05) showed that the autophagy pathway is impaired by HG concentrations. We concluded that visfatin can sufficiently decrease neural damage caused by ROS production and apoptosis under HG-induced toxicity. High glucose significantly increased the ROS level and apoptosis of diabetic PC12 cells;The autophagy pathway is impaired by high glucose;Nampt/PBEF/visfatin can significantly reduce neural damage caused by ROS production and apoptosis of diabetic PC12 cells. Diabetes mellitus is a metabolic disorder characterized by hyperglycemia resulting from a failure in insulin secretion, insulin action, or both. Visfatin (Nampt/PBEF) has insulin-mimetic effects. So far, no study has assessed its effects on diabetic neuropathy. Therefore, we examined the neuroprotective effects of visfatin on cell line 12 (PC12) against glucose-induced neurotoxicity. Based on the results, it was concluded that the Nampt/PBEF/visfatin can significantly reduce neural damage caused by production of reactive oxygen species and apoptosis of diabetic PC12 cell.

Molecular profiles and urinary biomarkers of upper tract urothelial carcinomas associated with aristolochic acid exposure.

Recurrent upper tract urothelial carcinomas (UTUCs) arise in the context of nephropathy linked to exposure to the herbal carcinogen aristolochic acid (AA). Here we delineated the molecular programs underlying UTUC tumorigenesis in patients from endemic aristolochic acid nephropathy (AAN) regions in Southern Europe. We applied an integrative multiomics analysis of UTUCs, corresponding unaffected tissues and of patient urines. Quantitative microRNA (miRNA) and messenger ribonucleic acid (mRNA) expression profiling, immunohistochemical analysis by tissue microarrays and exome and transcriptome sequencing were performed in UTUC and nontumor tissues. Urinary miRNAs of cases undergoing surgery were profiled before and after tumor resection. Ribonucleic acid (RNA) and protein levels were analyzed using appropriate statistical tests and trend assessment. Dedicated bioinformatic tools were used for analysis of pathways, mutational signatures and result visualization. The results delineate UTUC‐specific miRNA:mRNA networks comprising 89 miRNAs associated with 1,862 target mRNAs, involving deregulation of cell cycle, deoxyribonucleic acid (DNA) damage response, DNA repair, bladder cancer, oncogenes, tumor suppressors, chromatin structure regulators and developmental signaling pathways. Key UTUC‐specific transcripts were confirmed at the protein level. Exome and transcriptome sequencing of UTUCs revealed AA‐specific mutational signature SBS22, with 68% to 76% AA‐specific, deleterious mutations propagated at the transcript level, a possible basis for neoantigen formation and immunotherapy targeting. We next identified a signature of UTUC‐specific miRNAs consistently more abundant in the patients' urine prior to tumor resection, thereby defining biomarkers of tumor presence. The complex gene regulation programs of AAN‐associated UTUC tumors involve regulatory miRNAs prospectively applicable to noninvasive urine‐based screening of AAN patients for cancer presence and recurrence.

Prognostic and immune infiltrates for the Chromobox (CBX) protein family in human pancreatic adenocarcinoma.

Globally, pancreatic adenocarcinoma (PAAD) is among the most prevalent malignant tumors. The Chromobox (CBX) protein family is a vital component of epigenetic regulatory complexes that have vital biological roles. The biological functions, immune infiltration, expression levels, and the prognostic significance of CBX proteins in PAAD have not yet been established. Using bioinformatics tools, such as the Gene Expression Profiling Interactive Analysis (GEPIA), Oncomine, Kaplan-Meier plotter, GeneMANIA, cBioPortal, TIMER and R, we evaluated the prognostic importance, expression levels, gene alterations, risk factors, and immune cell infiltration levels of CBXs in PAAD patients. The expression levels of CBX3 in clinical-pathological samples were also confirmed by immunohistochemistry. In PAAD tissues, CBX1, CBX3, CBX5, and CBX8 expressions were high. High CBX1, CBX5, CBX6, and CBX7 levels were correlated with tumor stages. Elevated CBX2, CBX6, CBX7, and CBX8 messenger ribonucleic acid (mRNA) levels were markedly correlated with better overall survival (OS) outcomes for pancreatic cancer patients, while elevated CBX3 was markedly correlated with short OS outcomes. In particular, we have validated the differential expression of CBX3 on clinical specimens using immunohistochemical methods. A multivariate logistic analysis revealed that elevated mRNA expression levels of CBX3 and suppressed CBX8 levels were independently associated with short OS outcomes for pancreatic cancer patients. The roles of CBXs and their neighboring proteins are associated with a negatively regulated cell cycle, histone binding, polycomb group protein complexes, and the regulation of pluripotent stem cell signaling pathways. Additionally, CBX levels were found to be markedly associated with immune infiltrates, and found that the immune infiltration score of CBX3 was differentially expressed in cell lines such as CD8 T cells, NK cells, Mast cells and T helper cells. CBX3/8 is a potential marker for prognostic outcomes in PAAD patients.