UBE3A alleviates isoproterenol-induced cardiac hypertrophy through the inhibition of the TLR4/MMP-9 signaling pathway.

Cardiac hypertrophy is considered to be a leading factor in heart function-related deaths. In this study, we explored the potential mechanism underlying cardiac hypertrophy induced by isoproterenol. Our results showed that isoproterenol induced cardiac hypertrophy in AC16 cells, as reflected by the increased cell surface area and increased hypertrophic markers, which was accompanied by increased ubiquitin-protein ligase E3a (UBE3A) expression. Moreover, UBE3A knockdown by siRNAs accelerated cardiac hypertrophy, suggesting that increased UBE3A expression induced by isoproterenol might be a protective response and UBE3A might be a protective factor against cardiac hypertrophy. Our study also revealed that UBE3A knockdown increased the protein expression of the TLR4/MMP-9 pathway that has been shown to be associated with cardiac hypertrophy, which suggested that UBE3A-mediated protection is likely to be associated with the blockade of the TLR4/MMP-9 signaling pathway. UBE3A might be thus a potential target gene for the treatment of cardiac hypertrophy.

Expression and Prognostic Significance of p53 in Glioma Patients: A Meta-analysis.

Glioma is a brain tumor deriving from the neoplastic glial cells or neuroglia. Due to its resistance to anticancer drugs and different disease progress of individuals, patients with high-grade glioma are difficult to completely cure, leading to a poor prognosis and low overall survival. Therefore, there is an urgent need to look for prognostic and diagnostic indicators that can predict glioma grades. P53 is one of the widely studied biomarkers in human glioma. The purpose of this study was to comprehensively evaluate the significance of p53 expression in glioma grades and overall survival. We searched commonly used electronic databases to retrieve related articles of p53 expression in glioma. Overall, a total of 21 studies including 1322 glioma patients were finally screened out. We observed that the frequency of p53 immuno-positivity was higher in high-grade patients than that in low-grade category (63.8 vs. 41.6 %), and our statistic analysis indicated that p53 expression was associated with pathological grade of glioma (OR 2.93, 95 % CI 1.87-4.60, P < 0.00001). This significant correction was also found in 1-, 3- and 5-year overall survival. However, no positive relationship was found between age, sex, tumor size and p53 expression in patients with glioma. In conclusion, our results suggested that p53 immunohistochemical expression might have an effective usefulness in predicting the prognosis in patients with glioma.

Explore the alterations of downstream molecular pathways caused by ARID1A mutation/knockout in human endometrial cancer cells.

PURPOSE: As one of the most common gynecologic malignancies, endometrial cancer (EC) is driven by multiple genetic alterations that may be targeted for treatments. AT-rich interaction domain 1A (ARID1A) gene mutations were reported as early events in endometrial carcinogenesis. METHODS: To explore the alterations of downstream molecular pathways caused by ARID1A mutations and the associated therapeutic implications, we edited ARID1A gene in human endometrial cancer cell line Ishikawa using the Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-Associated Proteins (CRISPR/Cas9) technology. We successfully constructed a stable Ishikawa cell line with a confirmed 10 bp deletion on the ARID1A gene, which resulted in a code-shift mutation and gene knockout. RESULTS: Compared with unedited wild-type cells, ARID1A knockout (KO) led to reduced apoptosis, accelerated transformation from G0/G1 to S phase, and enhanced cell proliferation. ARID1A deficiency would reduce the protein levels of p21, caspase 7, and caspase 9 in Ishikawa endometrial cancer cells compared with the wild-type cells. In addition, ARID1A KO resulted in high levels of microsatellite instability (MSI-H). Moreover, transcriptomic analyses showed that ARID1A KO can lead to activated phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling. Furthermore, experimental analyses demonstrated that ARID1A KO cells had reduced expression of genetic instability-associated markers mutL homologue 1 (MLH1) and progesterone receptor B (PR) and increased p-Akt expression. CONCLUSION: These findings support further exploration of ARID1A as a therapeutic target for EC and provide insight into developing more effective treatments in EC, such as the combinatory use of immune checkpoint inhibitors.

Association between HO­1 gene promoter polymorphisms and diseases (Review).

Heme oxygenase­1 (HO­1) is an inducible cytoprotective enzyme that degrades heme into free iron, carbon monoxide and biliverdin, which is then rapidly converted into bilirubin. These degradation products serve an important role in the regulation of inflammation, oxidative stress and apoptosis. While the expression level of HO­1 is typically low in most cells, it may be highly expressed when induced by a variety of stimulating factors, a process that contributes to the regulation of cell homeostasis. In the 5'­non­coding region of the HO­1 gene, there are two polymorphic sites, namely the (GT)n dinucleotide and T(­413)A single nucleotide polymorphism sites, which regulate the transcriptional activity of HO­1. These polymorphisms have been shown to be closely associated with the occurrence and progression of numerous diseases, including cardiovascular, pulmonary, liver and kidney, various types of cancer and viral diseases. The present article reviews the progress that has been made in research on the association between the two types of polymorphisms and these diseases, which is expected to provide novel strategies for the diagnosis, treatment and prognosis of various diseases.

Recovery of human gut microbiota genomes with third-generation sequencing.

Human gut microbiota modulates normal physiological functions, such as maintenance of barrier homeostasis and modulation of metabolism, as well as various chronic diseases including type 2 diabetes and gastrointestinal cancer. Despite decades of research, the composition of the gut microbiota remains poorly understood. Here, we established an effective extraction method to obtain high quality gut microbiota genomes, and analyzed them with third-generation sequencing technology. We acquired a large quantity of data from each sample and assembled large numbers of reliable contigs. With this approach, we constructed tens of completed bacterial genomes in which there were several new bacteria species. We also identified a new conditional pathogen, Enterococcus tongjius, which is a member of Enterococci. This work provided a novel and reliable approach to recover gut microbiota genomes, facilitating the discovery of new bacteria species and furthering our understanding of the microbiome that underlies human health and diseases.

Pyrroloquinoline quinone attenuates isoproterenol hydrochloride­induced cardiac hypertrophy in AC16 cells by inhibiting the NF­κB signaling pathway.

Pyrroloquinoline quinone (PQQ) is a naturally occurring redox co­factor that functions as an essential nutrient and antioxidant, and has been reported to exert potent anti­inflammatory effects. However, the therapeutic potential of PQQ for isoproterenol hydrochloride (Iso)­induced cardiac hypertrophy has not yet been explored, at least to the best of our knowledge. In the present study, the anti­inflammatory effects of PQQ were investigated in Iso­treated AC16 cells, a myocardial injury cellular model characterized by an increase in the apparent surface area of the cells and the activation of intracellular cardiac hypertrophy­associated proteins. The results revealed that pre­treatment with PQQ significantly inhibited the expression of cardiac hypertrophy marker proteins, such as atrial natriuretic peptide, brain natriuretic peptide and ß­myosin heavy chain. PQQ also inhibited the activation of the nuclear factor (NF)­κB signaling pathway in Iso­treated AC16 cells, thus inhibiting the nuclear translocation of NF­κB and reducing the phosphorylation levels of p65. On the whole, the findings of this study suggest that PQQ may be a promising therapeutic agent for effectively reversing the progression of cardiac hypertrophy.

Low-abundance mutations in colorectal cancer patients and healthy adults.

Detecting low-abundance mutations is very important for cancer diagnosis and treatment. Here we describe an improved targeted sequencing analysis that dramatically increases sequencing depth. Seven colorectal cancer (CRC) patients and seven healthy adults were enrolled in this study. We examined genetic mutations in tissue samples from the central and peripheral regions of tumors from the CRC patients and in blood cells from the healthy adults. We observed that each CRC carried larger numbers of mutations more than previously estimated. These included numerous deletion mutations in the tumor tissue. While the cellular morphology in the surrounding normal colonic tissues was healthy, these cells also carried many mutations. Similarly, the blood cells from the healthy donors carried numerous mutations. These findings shed new light on the processes of tumorigenesis and aging, and also present a potentially effective method for detecting low-abundance mutations for cancer diagnosis and targeted treatments.

Pre-protective effect of polysaccharides purified from Hericium erinaceus against ethanol-induced gastric mucosal injury in rats.

The ß-glucan H6PC20 (Mw: 2390 kDa) and α-heteropolysaccharide HPB-3 (Mw: 15 kDa) were purified from the fruiting body of Hericium erinaceus according to the previous methods. Their gastroprotective activities and corresponding structure-activity relationship were studied in the ethanol-induced gastric ulcer model of rats. After intragastric administrated with H6PC20 and HPB-3 for 14 days, macroscopic and histological evaluation of gastric mucosa was improved significantly. The defense and repair factors (EGF, bFGF and PGE2) were increased, meanwhile, the inflammatory cytokines (IL-1ß and TNF-α) and MDA were reduced. These results indicated that H6PC20 and HPB-3 presented gastroprotective activities with the mechanism of activating repair and defense system, decreasing the inflammatory response and alleviating the oxidative injury. Furthermore, the structure-activity relationship showed that the macromolecular ß-glucan was better for repair and defense system, while the low weight molecular α-heteropolysaccharide focused on the anti-inflammatory effect. The polysaccharides purified from H. erinaceus can be developed as a potential gastroprotective ingredient for applications in pharmaceutical field.

Dgat2 reduces hepatocellular carcinoma malignancy via downregulation of cell cycle-related gene expression.

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide, mainly due to the absence of effective diagnostic biomarkers and therapeutic targets. Therefore, novel molecular targets are urgently needed, in order to formulate novel therapeutic approaches for this devastating disease. In the present study, we demonstrated that diacylglycerol acyltransferase 2 (Dgat2) was downregulated in human HCC tissues compared with in matched normal tissues. Furthermore, its high expression was significantly associated with longer survival. In addition, Dgat2 overexpression significantly suppressed HCC cell proliferation. in vivo studies, we revealed that the weight and volume of the tumors derived from Balb/c nude mice was markedly decreased when using HCC cells overexpressing Dgat2. Mechanism analysis demonstrated that cell cycle-related gene expressions were significantly downregulated in HCC cells overexpressing Dgat2. Taken together, these data suggest that Dgat2 is an important regulator of HCC cell proliferation, and could represent a potential anticancer target and diagnostic biomarker for HCC.

Long noncoding RNAs in autoimmune diseases.

With the completion of the human genome project and further development of high-throughput genomic technologies, interest in long noncoding RNAs (lncRNAs), which are defined as non-protein-coding RNAs at least 200 nucleotides in length, has strongly increased, and lncRNAs have become a major research direction. Increasing evidence demonstrates that lncRNAs are closely related to human growth and development and to disease occurrence via various mechanisms. lncRNAs also play crucial roles in the differentiation and activation of immune cells, and their relationships with human autoimmune diseases have received increasing attention. The development of biotechnology has led to the gradual discovery of many potential lncRNA functions. In this review, we discuss various lncRNAs that have been implicated in different human autoimmune diseases, focusing on their clinical applications as potential biomarkers and therapeutic targets in the pathologies of diverse human autoimmune diseases. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 468-475, 2019.

The whole transcriptome and proteome changes in the early stage of myocardial infarction.

As the most severe manifestation of coronary artery disease, myocardial infarction (MI) is a complex and multifactorial pathophysiologic process. However, the pathogenesis that underlies MI remains unclear. Here, we generated a MI mouse model by ligation of the proximal left anterior descending coronary artery. The transcriptome and proteome, at different time points after MI, were detected and analysed. Immune-related pathways, cell cycle-related pathways, and extracellular matrix remodelling-related pathways were significantly increased after MI. Not only innate immune cells but also adaptive immune cells participated in the early stage of MI. Proteins that functioned in blood agglutination, fibrinolysis, secretion, and immunity were significantly changed after MI. Nppa, Serpina3n, and Anxa1, three secreted proteins that can easily be detected in blood, were significantly changed after MI. Our discoveries not only reveal the molecular and cellular changes in MI but also identify potential candidate biomarkers of MI for clinical diagnosis or treatment.

Inhibition of DLC-1 gene expression by RNA interference in the colon cancer LoVo cell line.

DLC-1 (deleted in liver cancer-1) is a potential tumor suppressor gene, which is inactive in liver carcinogenesis. To observe the effects of DLC-1 gene expression on cell proliferation and migration in the human colon cancer cell line, RNAi Lipo-recombinant of the DLC-1 gene (pGCsil-DLC-1) was constructed and transduced into LoVo cells which are positive for DLC-1 gene expression. Results showed that the RNAi recombinant effectively inhibited the expression of the DLC-1 gene in LoVo cells. Additionally, our data showed decreased DLC-1 gene expression which resulted in the promotion of LoVo cell proliferation. Flow cytometry in cell cycle detection further indicated that the DLC-1 gene induced cell cycle arrest at G2/M and a cell migration assay confirmed that the knocking down of DLC-1 gene expression promotes LoVo cell migration. Our observations suggest that the DLC-1 gene is associated with LoVo cell proliferation, migration and cell cycle distribution. DLC-1 is a potential suppressor gene in the colon cancer LoVo cell line and may play an important role in colon cancer mechanisms.

Pharmacologic interventions for preventing delirium in adult patients after cardiac surgery: Protocol of a systematic review and network meta-analysis.

BACKGROUND: Delirium is common in adult patients undergoing cardiac surgery and related to a high morbidity and mortality. Although a variety of pharmacologic interventions have been applied in delirium prevention, there is still uncertainty concerning which drug is optimal. Thus, we plan to conduct a systematic review and network meta-analysis (NMA) of published studies to assess the efficacy and safety of pharmacologic interventions for preventing delirium among those patients. METHODS: A systematic literature search will be conducted in Embase, PubMed, and the Cochrane Library. The primary outcome will be the incidence of postoperative delirium. Secondary outcomes will include all-cause mortality and length of hospital or intensive care unit stay. A frequentist NMA will be conducted using Stata version 14.0. The inconsistency between direct and indirect comparisons will be evaluated using a node splitting method. In addition, surface under the cumulative ranking area will be used to evaluate superiority of different treatments. RESULTS: The findings of our review will be submitted to a peer-reviewed publication. CONCLUSION: Our study will generate convincing evidence regarding the effectiveness and safety of different pharmacologic interventions for delirium prevention in cardiac surgery patients.

The promising antitumour drug disulfiram inhibits viability and induces apoptosis in cardiomyocytes.

Disulfiram (DSF), widely used for treating alcohol abuse, is a promising antitumour drug that inhibits tumour cell viability, reverses cancer drug resistance and induces apoptosis. However, its potential side effects on cardiomyocytes remain unknown. This study demonstrated that DSF can not only inhibit cardiomyocyte viability and activity but also promote cell apoptosis. Furthermore, we revealed that cardiomyocytes were more sensitive to DSF than cancer cells. Moreover, the expression of STAT3, a key regulator of cardiomyocyte viability, was significantly down-regulated in cardiomyocytes treated with DSF. Finally, we also used experimental comparisons to indicate that PEG is a promising solvent for decreasing the adverse side effects of DSF, thereby expanding its potential range of clinical applications.

Mutation of the nm23-H1 gene has a non-dominant role in colorectal adenocarcinoma.

Nm23-H1 is a metastasis suppressor gene, which is has a reduced expression in patients with digestive system cancer. However, the mechanistic basis for the genetic instability remains unknown. To study the expression of the nm23-H1 gene in patients with colorectal cancer, polymerase chain reaction-single strand conformation polymorphism was used to analyze any point mutation, and immunohistochemistry was used to detect the expression of nm23-H1. Results revealed that all 63 specimens of Chinese human colorectal cancer tissues exhibit no point mutation. Among those 63 specimens, 19 (30%) exhibited positive immunostaining for the nm23-H1 protein and 44 (70%) exhibited negative immunostaining. These observations suggested that the protein and gene expression levels of nm23-H1 are reduced in colorectal cancer compared with the adjacent normal tissues, and the point mutation in the nm23-H1 gene is not the dominant cause of metastatic colorectal cancer.

USO1 promotes tumor progression via activating Erk pathway in multiple myeloma cells.

OBJECTIVE: This study aimed to explore the influence of USO1 on multiple myeloma (MM) cell proliferation and apoptosis and the related molecular mechanism. METHODS: The expression of USO1 and MIF in MM tissues and cells, normal bone marrow tissues and cells were determined by qRT-PCR and western blot assay. The cell proliferation and apoptosis of MM cells before and after knockdown of USO1 were determined by MTT assay and flow cytometry, respectively. Before and after knockdown of USO1, the expression of the proliferation-related genes cyclin D1, Mcm2 and PCNA in MM cells was determined by qRT-PCR and western blot assay. The protein level of p-Erk1/2 and MIF was determined by western blot assay and ELISA, respectively. RESULTS: The expression levels of USO1 and MIF in MM tissues and cells were much higher than those in normal bone marrow tissues and cells. Knockdown of USO1 resulted in the inhibited ability of cell proliferation and induced cell apoptosis. The expression of cyclin D1, Mcm2, PCNA and p-Erk1/2 decreased significantly after knockdown of USO1 as well as the decreased MIF secretion. CONCLUSION: USO1 gene may be a promising target for the therapy of human MM and its diagnosis marker.

Promoter methylation of the DLC­1 gene and its inhibitory effect on human colon cancer.

Deleted in liver cancer­1 (DLC­1), a candidate tumor suppressor gene which is inactive in liver carcinogenesis, is located at 8p21.3, where deletions are frequently found in several types of human cancer. Promoter hypermethylation is an epigenetic mechanism leading to silencing of the gene expression, which may be the primary cause for the absence of DLC­1. We investigated the expression of the DLC­1 gene and the methylation of the DLC­1 gene in colon cancer cell lines (Caco­2, LoVo and HT­29). The data showed that reduced or undetectable levels of DLC­1 mRNA were found in HT­29 by reverse transcription-polymerase chain reaction (RT­PCR). By contrast, the DLC­1 gene was significantly expressed in Caco­2 and LoVo cells. These findings were in agreement with the data obtained from western blot analysis. To further determine whether aberrant methylation is a contributing factor to transcriptional inactivation of DLC­1 in HT­29, the methylation of promoter was examined using methylation­specific PCR and sodium bisulfite genomic sequencing in LoVo and HT­29 cells, which suggests that promoter hypermethylation accounts for silencing of the DLC­1 gene in HT­29 cells. Since DLC­1 is a candidate tumor suppressor gene, we sought to determine whether DLC­1 expression is associated with cell proliferation in colon cancer cell lines. RNA interference techniques were adopted to inhibit DLC­1 expression in the LoVo cell line and resulted in inhibition of cell growth and reduced colony formation. Collectively, our observations suggest that hypermethylation is responsible for abrogating the function of the DLC­1 gene in colon cancer and indicate a role of DLC­1 in colon carcinogenesis.

Inhibition of nm23-H1 gene expression in chronic myelogenous leukemia cells.

For solid tumors of a malignant origin, the expression of the nm23-H1 gene is a positive prognostic factor. However, for chronic myeloid leukemia (CML), the prognostic role of nm23-H1 gene expression is unknown. The present study investigated the impact of nm23-H1 gene expression on the proliferation and migration of the CML K562 cell line to elucidate the association between nm23-H1 gene expression and CML cell survival. An RNAi lipo-recombinant plasmid of the nm23-H1 gene (pGCsi-nm23-H1) was constructed and transfected into the K562 cells. RT-PCR and western blotting were used to detect nm23-H1 mRNA and protein expression, respectively. The anchorage-independent growth ability of the transfected cells was observed in soft agar culture and the ability of the K562 cells to migrate was determined using a Transwell assay. Following the successful construction and transfection of the pGCsi-nm23-H1 plasmid into the K562 cells, nm23-H1 mRNA and protein expression levels were significantly lower compared with the control group. The stably-transfected pGCsi-nm23-H1 K562 cells exhibited a markedly increased ability to form colonies and the number and sizes of the colonies were significantly increased compared with those of the control. In vitro, the cells migrated through a Matrigel-coated membrane during incubation for 20 h. The Transwell assay revealed that the quantitative number of pGCsi-nm23-H1 K562 cells that migrated into the lower compartment of the invasion chamber was markedly increased compared with the control. In conclusion, nm23-H1 gene expression may inhibit K562 cell proliferation and migration. nm23-H1 may be a cancer suppressor gene and play a significant role in inhibiting the survival of CML cells.

Restoration of DLC1 gene inhibits proliferation and migration of human colon cancer HT29 cells.

DLC1 (deleted in liver cancer-1) is a new candidate tumor suppressor gene, which is inactive in various types of human cancers including colon cancer. To study the function of DLC1, we constructed a pcDNA3.1 vector containing the DLC1 gene and transfected it into HT29 colon cancer cells that were deficient in DLC1 expression. The restoration of DLC1 expression in HT29 cells significantly inhibited cell proliferation and migration. Flow cytometry showed that DLC1 transfection into HT29 cells induced apoptosis and that the cell cycle was arrested at S-phase. Additionally, cyclinD1 mRNA and protein expression were down-regulated while p21 expression was increased in pcDNA3.1-DLC1-HT29 cells compared to wild HT29 cells. These results confirm the role of DLC1 gene as a tumor suppressor, which may be manifested by regulation of p21 and cyclinDl. The DLC1 gene has a potential therapeutic role in inhibiting the development of colon cancer.