Mechanism of lncRNA SNHG16 on kidney clear cell carcinoma cells by targeting miR-506-3p/ETS1/RAS/ERK molecular axis.

Objective: This study aimed to investigate the mechanism of long noncoding ribonucleic acid (lncRNA) SNHG16 on kidney clear cell carcinoma (KIRC) cells by targeting miR-506-3p/ETS proto-oncogene 1, transcription factor (ETS1)/RAS/Extracellular regulated protein kinases (ERK) molecular axis, thus to provide reference for clinical diagnosis and treatment of KIRC in the future. Methods: Thirty-six patients with KIRC were enrolled in this study, and their carcinoma tissues and adjacent tissues were obtained for the detection of SNHG16/miR-506-3p/ETS1/RAS/ERK expression. Then, over-expressed SNHG16 plasmid and silenced plasmid were transfected into KIRC cells to observe the changes of their biological behavior. Results: SNHG16 and ETS1 were highly expressed while miR-506- 3p was low expressed in KIRC tissues; the RAS/ERK signaling pathway was significantly activated in KIRC tissues (P < 0.05). After SNHG16 silence, KIRC cells showed decreased proliferation, invasion and migration capabilities and increased apoptosis rate; correspondingly, increase in SNHG16 expression achieved opposite results (P < 0.05). Finally, in the rescue experiment, the effects of elevated SNHG16 on KIRC cells were reversed by simultaneous increase in miR-506-3p, and the effects of miR-506-3p were reversed by ETS1. Activation of the RAS/ERK pathway had the same effect as increase in ETS1, which further worsened the malignancy of KIRC. After miR-506-3p increase and ETS1 silence, the RAS/ERK signaling pathway was inhibited (P < 0.05). At last, the rescue experiment (co-transfection) confirmed that the effect of SNHG16 on KIRC cells is achieved via the miR-506-3p/ETS1/RAS/ERK molecular axis. Conclusion: SNHG16 regulates the biological behavior of KIRC cells by targeting the miR-506-3p/ETS1/RAS/ERK molecular axis.

Scientometric Analysis and Research Mapping Knowledge of Coconut Fibers in Concrete.

Biodegradable materials are appropriate for the environment and are gaining immense attention worldwide. The mechanical properties (such as elongation at break, density, and failure strain) of some natural fibers (such as Coir, Hemp, Jute, Ramie, and Sisal) are comparable with those of some synthetic fibers (such as E glass, aramid, or Kevlar). However, the toughness of coconut fibers is comparatively more than other natural fibers. Numerous studies suggest coconut fibers perform better to improve the concrete mechanical properties. However, the knowledge is dispersed, making it difficult for anyone to evaluate the compatibility of coconut fibers in concrete. This study aims to perform a scientometric review of coconut fiber applications in cementitious concrete to discover the various aspects of the literature. The typical conventional review studies are somehow limited in terms of their capacity for linking different literature elements entirely and precisely. Science mapping, co-occurrence, and co-citation are among a few primary challenging points in research at advanced levels. The highly innovative authors/researchers famous for citations, the sources having the highest number of articles, domains that are actively involved, and co-occurrences of keywords in the research on coconut-fiber-reinforced cementitious concrete are explored during the analysis. The bibliometric database with 235 published research studies, which are taken from the Scopus dataset, are analyzed using the VOSviewer application. This research will assist researchers in the development of joint ventures in addition to sharing novel approaches and ideas with the help of a statistical and graphical description of researchers and countries/regions that are contributing. In addition, the applicability of coconut fiber in concrete is explored for mechanical properties considering the literature, and this will benefit new researchers for its use in concrete.

A pilot retrospective study of comprehensive nursing care on psychological disorder in colorectal cancer undergoing chemotherapy.

BACKGROUND: This pilot study retrospectively evaluated the effects of comprehensive nursing care (CNC) on psychological disorders in patients with colorectal cancer (CC) undergoing chemotherapy. METHODS: This study analyzed 70 eligible patients' case records of CC undergoing chemotherapy. These records were allocated to a treatment group (n = 35) or a control group (n = 35). All 70 patients in both groups received routine nursing care. In addition, 35 patients in the treatment group also received CNC. The primary outcomes were anxiety, as measured by Self-rating Anxiety Scale, and depression, as assessed by Self-rating Depression Scale. The secondary outcomes were quality of life, as measured by The 36-Item Short Form Health Survey, and adverse events. All outcome data were analyzed before and 3-month after treatment. RESULTS: At 3-month after treatment, the patients in the treatment group had better outcomes in the Self-rating Anxiety Scale (P<0.01), Self-rating Depression Scale (P<0.01), and The 36-Item Short Form Health Survey (social function, P = .04; emotional role, P = 0.03) than those in the control group. With regard to safety, no treatment-related adverse events were recorded in either group. CONCLUSION: The findings of this pilot retrospective study showed promising effects of CNC on psychological disorders and quality of life in patients with CC undergoing chemotherapy. However, more high-quality clinical trials are required to confirm these findings.

CT Image Features Based on the Reconstruction Algorithm for Continuous Blood Purification Combined with Nursing Intervention in the Treatment of Severe Acute Pancreatitis.

The aim of the study was to explore the CT images of the iterative reconstruction algorithm to evaluate the curative effect of continuous blood purification combined with nursing intervention in the treatment of severe acute pancreatitis (SAP). A total of 100 patients with SAP treated by the bedside continuous venous hemofiltration purification method in a hospital were selected. The control group (n = 50) was given a routine treatment, and the observation group (n = 50) was treated with the continuous blood filtration mode for blood purification based on the routine treatment. In the CT image scanning of periodontitis patients, the iterative reconstruction algorithm was introduced to reduce image noise, and the CT values under the algorithm were statistically analyzed. The results showed that IL-1, IL-6, and IL-8 after treatment were significantly lower than those before treatment (P < 0.05). The symptoms effectively improved with continuous blood purification combined with nursing intervention in patients with SAP. After the use of the iterative reconstruction algorithm, the image quality, image information, and image MSE significantly improved. The image noise with 50% dose reduction was the lowest, but the reconstruction algorithm improved the low contrast resolution (P < 0.05). CT images based on the reconstruction algorithm can clearly display the lesion characteristics of the patients, and the reconstruction algorithm is feasible to improve the spatial resolution of CT images.

CT Image Feature under Intelligent Algorithm in the Evaluation of Continuous Blood Purification in the Treatment and Nursing of Pulmonary Infection-Caused Severe Sepsis.

This study was to explore the CT image features based on intelligent algorithm to evaluate continuous blood purification in the treatment of severe sepsis caused by pulmonary infection and nursing. 50 patients in the hospital were selected as the research objects. Convolutional neural network algorithm was used to segment CT images of severe sepsis caused by pulmonary infection. They were randomly divided into 25 cases of experimental group and 25 cases of control group. The experimental group was given continuous blood purification treatment, combined with comprehensive nursing. The control group was given routine treatment and basic nursing. Fasting plasma glucose (FPG) and fasting insulin (FIN), interleukin-6 (IL-6), tumor necrosis factor (TNF-α), high-sensitivity c-reactive protein (hs-CRP) levels, CD3 +, CD4 +, CD4 +/CD8 + levels, ICU monitoring time, malnutrition inflammation score (MIS), and incidence of adverse events were compared between the two groups before and after treatment. There was no difference in FPG and FIN between the two groups before treatment. After treatment, the FPG and FIN of the experimental group were lower than those of the control group, and there was statistical significance (P < 0.05). There was no difference in IL-6, TNF-α, and hs-CRP between the two groups before treatment. After treatment, IL-6, TNF-α, and hs-CRP in the experimental group were lower than those in the control group. There was no difference in the percentage of CD3 +, CD4 +, and CD4 +/CD8 + between the two groups before treatment. After treatment, the CD3 +, CD4 +, and CD4 +/CD8 + in the experimental group were higher than those in the control group. The ICU monitoring time, MIS, and incidence of adverse events in the experimental group were lower than those in the control group (P > 0.05). Convolutional neural network algorithm can accurately identify and segment CT images of patients with severe sepsis, which has high clinical application value. Continuous blood purification therapy can effectively control blood glucose level, improve immune function, and reduce the content of inflammatory factors in patients with severe sepsis caused by pulmonary infection. Effective nursing measures can improve the therapeutic effect.

A metabolomic signature of FIGO stage I and II endometrial cancer.

Endometrial cancer (EC) is a malignant tumor of the female reproductive tract. Due to its rapid growth and invasiveness, EC is currently the only gynecological neoplasm with rising incidence and mortality rates. It is of great significance to explore the metabolomics signature of stage I and II EC for the diagnosis and treatment. A mass spectrometry-based untargeted metabolomics approach was used to explore preoperative serum metabolites in the normal and stage I and II EC patients. The metabolites were mapped to the Ingenuity pathway analysis (IPA) database to determine the potential biomarkers and metabolic pathways that differ between EC patients and healthy controls. The top analysis-ready molecules of upregulated D-glucose thiamine and downregulated cholesterol, arachidonic acid, palmitic acid, oleic acid, stearic acid, linoleic acid may be the most related metabolites. These potential biomarkers have essential functions in regulating vital metabolic pathways associated with stage I and II EC. Additionally, our pathway analysis revealed five significantly related pathways according to the metabolite differentials. Finally, the disease and function prediction of the initial pathway analysis suggested that small molecule biochemistry, lipid metabolism, and organismal injury and abnormalities were associated with EC cases. Over 25 metabolites were differentially expressed in stage I and II EC. In addition, the six most significant metabolites were related to stage I and stage II EC cases. Ingenuity pathway analysis revealed potential biomarkers and metabolic pathways revolved to EC. In this paper, candidate endogenous biomarkers were defined as the basis for disease diagnosis and individualized treatment monitoring and revealed the mechanism of EC occurrence and development.

Metabolite reanalysis revealed potential biomarkers for COVID-19: a potential link with immune response.

Aim: To understand the pathological progress of COVID-19 and to explore the potential biomarkers. Background: The COVID-19 pandemic is ongoing. There is metabolomics research about COVID-19 indicating the rich information of metabolomics is worthy of further data mining. Methods: We applied bioinformatics technology to reanalyze the published metabolomics data of COVID-19. Results: Benzoate, ß-alanine and 4-chlorobenzoic acid were first reported to be used as potential biomarkers to distinguish COVID-19 patients from healthy individuals; taurochenodeoxycholic acid 3-sulfate, glucuronate and N,N,N-trimethyl-alanylproline betaine TMAP are the top classifiers in the receiver operating characteristic curve of COVID-severe and COVID-nonsevere patients. Conclusion: These unique metabolites suggest an underlying immunoregulatory treatment strategy for COVID-19.

SNHG16 promotes cell proliferation and inhibits cell apoptosis via regulation of the miR-1303-p/STARD9 axis in clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is a common subtype of renal cell carcinoma (RCC) and causes many deaths. Numerous medical studies have suggested that long noncoding RNAs (lncRNAs) exert their biological functions on ccRCC. Herein, functions of lncRNA SNHG16 in ccRCC cells and the mechanism mediated by SNHG16 were investigated. The expression levels of SNHG16 and its downstream genes in ccRCC cells and RCC tissues were examined utilizing reverse transcription quantitative polymerase chain reaction analyses. Cell counting kit-8 and 5-Ethynyl-2'-deoxyuridine assays were performed to evaluate the proliferation of ccRCC cells, and flow cytometry analyses were employed to determine the apoptosis of ccRCC cells. Western blot analysis was applied to examine protein levels associated with cell proliferation and apoptosis. The combination between SNHG16 and miRNA as well as miRNA and its target gene were explored by luciferase reporter, RNA pull down, and RNA immunoprecipitation assays. The significant upregulation of SNHG16 was observed in RCC tissues and ccRCC cells. SNHG16 downregulation inhibited the proliferation and promoted the apoptosis of ccRCC cells. In addition, SNHG16 served as a competing endogenous RNA for miR-1301-3p, and STARD9 was a target gene of miR-1301-3p in ccRCC cells. SNHG16 upregulated STARD9 expression by binding with miR-1301-3p in ccRCC cells. Rescue assays validated that SNHG16 promoted ccRCC cell promotion and induced ccRCC cell apoptosis by upregulating STARD9 expression. In conclusions, SNHG16 promotes ccRCC cell proliferation and suppresses ccRCC cell apoptosis via interaction with miR-1301-3p to upregulate STARD9 expression in ccRCC cells.

The prognostic and predictive value of tumor infiltrating Macrophage and Neutrophil in patient with clear cell renal cell carcinoma: Tumor infiltrating lymphocytes in renal cell carcinoma.

Renal cell carcinoma (RCC) is the leading cancer affecting humans; however, the relationship between tumour-infiltrating lymphocytes (TILs) and patient prognosis in RCC is relatively unreported. This study aimed to investigate the relationships among factors (TIL, clinicopathological characteristics, and patient prognosis in RCC).This retrospective study evaluated 533 patients with clear cell renal cell carcinoma (ccRCC) deposited in the the Cancer Genome Atlas between 2004 and 2015. We downloaded immune cell type absolute fraction data for ccRCC patients from the Cancer Immunome Atlas database. The CIBERSORT method was used to transform RNA-sequencing data into microarray data for the cancer genome atlas -ccRCC samples for which microarray and RNA-sequencing data were available on the the Cancer Immunome Atlas website.The overall survival (OS) and disease free survival (DFS) analyses of ccRCC patients showed that M1 macrophages (OS, P = .00000134; DFS, P = .00958) and neutrophils (OS, P = .00000723; DFS, P = .0255) were significant. Age at diagnosis (P < .0001, c-index = 0.59), tumour stage (P < .0001, c-index = 0.667), stage (P < .0001, c-index = 0.729), neoplasm histological grade (P < .0001, c-index = 0.624), and haemoglobin level (P < .0001, c-index = 0.583) were independent predictors of OS. Similarly, the stage, haemoglobin level, and serum calcium level were independent predictors of DFS. There were significant correlations between the M1 macrophage fraction and tumour stage, stage, and neoplasm histological grade. Stage and neoplasm histological grade showed associations with the neutrophil fraction.The correlations between TILs and prognosis and clinicopathological characteristics in ccRCC were demonstrated. The prognosis of ccRCC patients may differ according to the TIL fractions.

Identification of the subtypes of gastric cancer based on DNA methylation and the prediction of prognosis.

BACKGROUND: Gastric cancer (GC) is a digestive system cancer with a high mortality rate globally. Previous experiences and studies have provided clinicians with ample evidence to diagnose and treat patients with reasonable therapeutic options. However, there remains a need for sensitive biomarkers that can provide clues for early diagnosis and prognosis assessment. RESULTS: We found 610 independent prognosis-related 5'-cytosine-phosphate-guanine-3' (CpG) sites (P < 0.05) among 21,121 sites in the training samples. We divided the GC samples into seven clusters based on the selected 610 sites. Cluster 6 had relatively higher methylation levels and high survival rates than the other six clusters. A prognostic risk model was constructed using the significantly altered CpG sites in cluster 6 (P < 0.05). This model could distinguish high-risk GC patients from low-risk groups efficiently with the area under the receiver operating characteristic curve of 0.92. Risk assessment showed that the high-risk patients had poorer prognosis than the low-risk patients. The methylation levels of the selected sites in the established model decreased as the risk scores increased. This model had been validated in testing group and its effectiveness was confirmed. Corresponding genes of the independent prognosis-associated CpGs were identified, they were enriched in several pathways such as pathways in cancer and gastric cancer. Among all of the genes, the transcript level of transforming growth factor ß2 (TGFß2) was changed in different tumor stages, T categories, grades, and patients' survival states, and up-regulated in patients with GC compared with the normal. It was included in the pathways as pathways in cancer, hepatocellular carcinoma or gastric cancer. The methylation site located on the promoter of TGFß2 was cg11976166. CONCLUSIONS: This is the first study to separate GC into different molecular subtypes based on the CpG sites using a large number of samples. We constructed an effective prognosis risk model that can identify high-risk GC patients. The key CpGs sites or their corresponding genes such as TGFß2 identified in this research can provide new clues that will enable gastroenterologists to make diagnosis or personalized prognosis assessments and better understand this disease.

Increased expression of YTHDF1 and HNRNPA2B1 as potent biomarkers for melanoma: a systematic analysis.

BACKGROUND: The incidence and mortality of melanoma is increasing around the world. To deeply explain the mechanism insight into it, we conducted a systematic analysis to examine the levels of regulatory genes of the common RNA epigenetic modification-N6-methyladenosine (m6A) in patients with melanoma compared by the healthy. METHODS: We analyzed the expression of m6A Eraser, Writer, and Reader genes based on publicly available datasets on Oncomine and validated the results with a gene expression omnibus dataset. Hub genes were identified with Cytohubba and the frequency of copy number alterations was analyzed with the cBioPortal tool. RESULTS: The results revealed the up-regulation of YTHDF1 and HNRNPA2B1 in melanoma. Combining the two genes improved the efficacy in diagnosing melanoma by about 10% compared to each gene alone. Hub genes identified with four analysis methods were compared and the overlapping genes were selected. These genes were enriched in several gene ontology terms. Genes related to p53-signaling consisted of CDK2, CDK1, RRM2, CCNB1, and CHEK1. All five genes were positively correlated with either YTHDF1 or HNRNPA2B1, suggesting that both genes may affect m6A modification by the five genes, further up-regulating their expression and facilitate their roles in inhibiting p53 to suppress tumorigenesis. We also observed major mutations in YTHDF1 and HNRNPA2B1 that led to their amplification in melanoma. Significant differences were observed in the clinical characteristics of patients with altered and unaltered m6A regulatory genes such as tumor stage and treatment response. CONCLUSIONS: We, for the first time, identified a combination of m6A regulatory genes to diagnose melanoma. We also analyzed m6A-related genes more comprehensively based on systematic complete data. We found that YTHDF1 and HNRNPA2B1 were altered in melanoma and might influence the development of the disease through signaling pathways such as p53.

Increased CD200 levels in peripheral blood mononuclear cells of patients with primary Sjögren's syndrome.

OBJECTIVES: Primary Sjögren's syndrome (pSS) is a chronic autoimmune disease with an unknown etiology. CD200 is associated with many autoimmune diseases, but little is known about its role in pSS. This study aims to correlate the expression of CD200 with pSS and evaluate its significance. METHODS: Plasma CD200, CD200R, and interleukin (IL)-17 levels were measured and analyzed by enzyme-linked immunosorbent assay. Messenger RNA levels of CD200 and CD200R in peripheral blood mononuclear cells (PBMCs) were quantified by quantitative real-time polymerase chain reaction (RT-qPCR). Following pretreatment of CD200-Fc, the protein levels of IL-17A were measured in PBMCs from patients and healthy controls. RESULTS: Results showed that, compared to CD200 in healthy controls, the relative levels in PBMCs from pSS were greater than 2-fold. In addition, CD200 levels in plasma positively correlated with IL-17 levels, as well as between plasma CD200 and pSS activity indexes (including immunoglobulin G and European League Against Rheumatism SS Disease Activity Index). While CD200R levels were significantly decreased in pSS patients, no correlation could be found. Furthermore, the protein level of IL-17 decreased after pretreatment of CD200-Fc in PBMCs from pSS patients. CONCLUSION: Our results suggested that the CD200/CD200R pathway is involved in pSS pathogenesis. It is hypothesized that regulation of IL-17 expression affects Th17 differentiation. This newly discovered pathway could give rise to a novel targeted therapy for pSS.

Comparison of 3-dimensional laparoscopy and conventional laparoscopy in the treatment of complex renal tumor with partial nephrectomy: A propensity score-matching analysis.

To compare the efficacies of 3-dimensional laparoscopic partial nephrectomy and conventional laparoscopic partial nephrectomy for complex renal tumors. The complex renal tumors was defined as Preoperative Aspects and Dimensions Used for an anatomical (PADAU) ≥10, including some cT1b tumors.This was a retrospective analysis of patients with local complex renal tumors who presented to our hospital from January 2014 to January 2018. All patients were managed with laparoscopic partial nephrectomy (LPN) or 3-dimensional partial nephrectomy (3DLPN).There were 48 patients in the LPN group and 60 in the 3DLPN group. In the matched groups, demographic and tumor characteristics including Charlson Comorbidity Index, PADUA, based on the preoperative images, were similar. By contrast, 3DLPN achieved better results in terms of warm ischemia time (19 vs 27 minutes), operation time (105 vs 128 minutes), postoperative complications (14.9% vs 23.4%), and marginal width (0.6 cm vs 0.4 cm). We found statistically significant differences in terms of length of stay, estimated blood loss (EBL), positive surgical margin (PSM), and conversion to open or radical nephrectomy (RN). Median follow-up time was 17 and 18.5 months for the LPN and 3DLPN groups, respectively. The recovery of renal function (% change eGFR, 0 vs -8.7) was significantly different between the 3DLPN and LPN groups, whereas 12-month recurrence-free survival did not differ.Both 3-dimensional laparoscopic nephron-sparing nephrectomy and conventional laparoscopic partial nephrectomy are safe, effective, and acceptable approaches to treating complex renal tumors, while the former may facilitate tumor resection and renorrhaphy for challenging cases, offering a minimally invasive surgical option for patients who may otherwise require open surgery.

Aberrant IL-35 levels in patients with primary Sjogren's syndrome.

OBJECTIVE: IL-35 is a newly discovered immunoregulatory cytokine that possesses the ability to inhibit CD4 + effector T cells and alleviate autoimmune diseases. The objective of this study was to investigate IL-35 levels in patients with primary Sjogren's syndrome (pSS) and explore the roles of IL-35 in the pathogenesis of pSS. METHODS: Thirty-four hospitalized patients with pSS were recruited, and 34 volunteers were enrolled as healthy controls. An ELISA was adopted to measure plasma IL-35 levels. The levels of P35 and EBI3 mRNAs in peripheral blood mononuclear cells (PBMCs) were determined using real-time quantitative PCR. The percentage of CD4 + EBI3 + T cells and CD19 + EBI3 + B cells was analysed using flow cytometry. Correlations between IL-35 levels, P35 and EBI3 mRNAs, numbers of CD4 + EBI3 + T cells, CD19 + EBI3 + B cells and clinical parameters were analysed. RESULTS: Significantly lower plasma IL-35 levels, P35 and EBI3 mRNA levels, and percentages of CD4 + EBI3 + T cells but increased percentages of CD19 + EBI3 + B cells were observed in patients with pSS than in healthy controls. IL-35 levels, EBI3 mRNA expression and the percentage of CD4 + EBI3 + T cells exhibited negative correlations with the ESSDAI score, whereas levels of the IL-35 protein and EBI3 mRNA were negatively correlated with the ESR. Patients who were positive for anti-SSB antibodies presented with lower IL-35 levels and percentages of CD4 + EBI3 + T cells. CONCLUSIONS: Based on these results, a decrease in the IL-35 levels may play an important role in the pathogenesis of pSS. IL-35 may act as a potential therapeutic agent against inflammation in patients with pSS.

Magnetic-bioluminescent-nanoliposomes for ultrasensitive and portable detection of protein biomarkers in blood.

Protein biomarkers play an important role in the diagnosis and treatment of disease. Herein, we report an ultrasensitive magnetic-bioluminescent-nanoliposome based technique using a portable ATP luminometer for the point-of-care testing (POCT) of protein biomarkers in blood. In this study, we use alpha-fetoprotein (AFP, a protein biomarker associated with hepatocellular carcinoma) as a model protein. The bioluminescent nanoliposomes conjugated to magnetic particles (LBM) enabled target protein capture, isolation and detection. In this assay, we use a portable magnetic bead separation pen to simplify the steps. The output RLUs (relative light units) had a linear correlation with AFP concentration between 0.05 and 1000 ng/mL, with a limit of detection of 0.016 ng/mL. Our LBM assay for AFP based on the portable luminometer exhibited high specificity for AFP, with no cross-reactivity with other proteins tested at 25 ng/mL. Forty clinical samples (twenty AFP positive and twenty AFP negative) were tested by the LBM assay, and the results were in good agreement with those determined by electrochemiluminescence, with relative deviations of less than 10%. The successful application of magnetic-bioluminescent-nanoliposomes with a portable ATP luminometer system to detect protein biomarkers in blood has opened a new avenue for biomarker testing. Thus, our LBM assay holds great potential as a POCT assay for use in clinical diagnostics.

Abnormal Expression of Long Noncoding RNAs in Primary Immune Thrombocytopenia: A Microarray Related Study.

BACKGROUND/AIMS: Long noncoding RNAs (lncRNAs) are important regulators of biological processes and they contribute to the pathological developments of various diseases, including autoimmune diseases. To gain the further understanding, we estimate the expression of lncRNAs in primary immune thrombocytopenia (ITP). METHODS: In this study, microarray studies were performed to characterize expression profiles of various lncRNAs and mRNAs in blood samples collected from ITP patients. Quantitative real-time PCR (qRT-PCR) was performed to confirm the results, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and gene ontology analysis were used to provide functional annotations, co-expression network construction (CNC) analysis was made to reveal the relations between lncRNAs and their targeted genes. RESULTS: A total of 1177 and 632 lncRNAs were significantly up-regulated or down-regulated, respectively, in "newly diagnosed ITP" patients versus healthy individuals. In addition, 1182 genes and 737 genes were up-regulated or down-regulated, respectively, in "chronic recurrent ITP" patients versus healthy individuals. In a KEGG analysis, "TNF signaling pathway-Homo sapiens (human)" was a key result. In a gene ontology analysis, "Granulocyte macrophage colony-stimulating factor production (GO: 0032604, ontology: Biological process, P = 1.69577E-05)" and "coreceptor activity (GO: 0015026, ontology: molecular function, P = 4.67594E-06)" were the two most critical results. Data from qRT-PCR and receiver operating characteristic curves further demonstrated that ENST00000440492, ENST00000528366, NR_038920, and ENST00000552576 can efficiently distinguish different stages of ITP, especially NR_038920 and ENST00000528366. In a CNC analysis, four lncRNAs were emphasized, and NR_038920 and ENST00000528366 were both associated with proteins with important roles in autoimmune diseases. CONCLUSIONS: These results suggest that lncRNAs act through targeted genes to mediate their functions and to mediate their functions and affect the pathogenesis of ITP.

Clinical Significance of Decreased Interleukin-35 Expression in Patients with Psoriasis.

In psoriasis, a chronic, recurrent, inflammatory skin disease, CD4+T cells and their related cytokines play an important role in its pathogenesis. The role of interleukin (IL)-35, an immunosuppressive cytokine involved in many autoimmune diseases, is unclear in the pathogenesis of psoriasis. This study evaluated IL-35 expression and clinical significance in psoriasis. Protein and mRNA levels of specified markers were measured by enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction (qRT-PCR), respectively. Results showed that plasma IL-35 concentrations were lower in patients with psoriasis than in healthy individuals (Z = -6.525, P < .0001). Ebi3 and p35 showed lower mRNA levels in peripheral blood mononuclear cells from patients with psoriasis than in healthy individuals (Z = -5.078, P < .0001, Z = -2.609, P = .009, respectively). The areas under the receiver-operating characteristic (ROC) curves of IL-35, Ebi3, and p35 for patients with psoriasis versus the control were 0.86, 0.78, and 0.64, respectively. Pearson correlation analysis showed that plasma IL-35 expression negatively correlated with interferon-gamma, tumor necrosis factor-alpha, levels of IL-23, -17, and -22, or the Psoriasis Activity and Severity Index and positively correlated with levels of transforming growth factor beta and IL-10 levels in patients with psoriasis. Summarily, IL-35 might mediate psoriasis pathogenesis by influencing the expression of Th1/Th17/Treg -related cytokines and might be a putative target in monitoring or treating psoriasis.

Lower Plasma Levels of IL-35 in Patients with Primary Biliary Cirrhosis.

Primary biliary cirrhosis (PBC) is an autoimmune liver disease. Its histological characteristics, such as progressive intrahepatic bile duct destruction, cholestasis, and liver cirrhosis, are caused by the body's autoimmune disorders. Interleukin (IL)-35 has two subunits (p35 and Ebi3) and is a member of the IL-12 family of heterodimeric cytokines. IL-35 has immunosuppressive functions and plays an important role in many autoimmune diseases. In this study, we compared plasma levels of IL-35 and relative mRNA expression levels of p35 and Ebi3 in peripheral blood mononuclear cells (PBMCs) from 70 PBC patients and 70 healthy individuals. The results showed that the relative expression levels of Ebi3 mRNA were lower in PBMCs from PBC patients than in PBMCs from healthy individuals, whereas the levels of p35 mRNA were similar in both groups. Plasma IL-35 concentrations were lower in patients with PBC than in healthy individuals. Plasma levels were higher in PBC patients at an advanced stage compared to patients at an early stage. Variable plasma levels with different stages were also found in transforming growth factor beta (TGF-ß), which is mainly produced by regulatory T cells (Tregs). IL-35 and TGF-ß levels were positively correlated with each other, and IL-35 was capable of promoting the inhibitory functions of Tregs in PBC patients at both the early and late stages of disease. Lower plasma IL-35 levels were accompanied by higher levels of typical clinical parameters, such as alkaline phosphatase, or of proinflammatory cytokines, such as interferon-gamma (IFN-γ), in PBC patients (P < 0.05 for each). We propose that IL-35 may be involved in the pathogenesis of PBC and could be a potential biomarker for diagnosing this disease.

Differential Expression of MiR-106b-5p and MiR-200c-3p in Newly Diagnosed Versus Chronic Primary Immune Thrombocytopenia Patients Based on Systematic Analysis.

BACKGROUND/AIMS: MicroRNAs (miRNAs) have been described to have important roles in primary immune thrombocytopenia (ITP). To gain additional understanding, we have now further evaluated the involvement of miRNAs in ITP. METHODS: Microarray experiments were performed to examine the expression profiles of miRNAs and mRNAs in samples from subjects with newly diagnosed ITP (G1), chronic ITP (G2), and normal controls. The systematic Pipeline of Outlier MicroRNA Analysis framework was applied to identify key miRNAs expressed in the G1 and G2 samples. Quantitative PCR and receiver operator characteristic curves were used to confirm the performance of key miRNAs. RESULTS: Compared with normal controls, 14 miRNAs (12 over-expressed and 2 under-expressed) and 7 over-expressed miRNAs were identified as key in G1 and G2 samples, respectively. miR-106b-5p, miR-200c-3p, and miR-92a-3p exhibited significantly different expression profiles among the groups. In particular, miR-106b-5p and miR-200c-3p were expressed at higher levels in patients with ITP compared to the normal controls. Furthermore, these two miRNAs expressions were even higher in patients with chronic ITP. CONCLUSION: MiR-106b-5p and miR-200c-3p may represent valuable biomarkers of ITP, although further studies are needed to confirm and assess the value of these potential biomarkers at various stages of ITP.

Nano-biotinylated liposome-based immunoassay for the ultrasensitive detection of protein biomarker in urine.

With the development of proteomics and the continuous discovery of biomarkers of trace proteins, it is important to accurately quantify low abundance protein, especially in urine for clinical diagnostics. In this paper, we reported a novel nano-biotinylated liposome-based immuno-loop-mediated isothermal amplification (LI-LAMP) for the ultrasensitive detection of REG1A (a biomarker for pancreatic ductal adenocarcinoma (PDAC) in urine) with high specificity. The detection range was 1µg/mL to 1fg/mL, with a detection limit of 1fg/mL, and no cross-reactivity was observed to occur in this assay. Compared with the amount of REG1A added, REG1A recovery using this method was 130% and 89%. Detection of REG1A concentrations using the LI-LAMP assay from real samples were in good agreement with those determined using ELISA, and relative deviations were not more than 10%. LI-LAMP shows good potential as a clinical diagnostic assay.