Circulating micrornas as potential diagnostic biomarkers for cervical intraepithelial neoplasia and cervical cancer: a systematic review and meta-analysis.

BACKGROUND: Cervical cancer is a prevalent malignancy of the female reproductive system. Cervical intraepithelial neoplasia (CIN) is a precursor lesion for CC. Various studies have examined circulating microRNAs (miRNAs) as potential early diagnostic markers for CC and CIN. However, the findings have been inconclusive. Therefore, it is necessary to evaluate the diagnostic accuracy and identify potential sources of variability among these studies. METHODS: The PubMed, Cochrane Library, Embase, and Web of Science databases were searched to identify relevant literature. Then, Stata 14.0 was utilized to calculate summary estimates for diagnostic parameters, including sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and area under the summary receiver operating characteristic (ROC). To scrutinize the heterogeneity, the Cochran-Q test and I2 statistic were utilized. As significant heterogeneity was observed, the random effects model was chosen. To explore potential sources of the heterogeneity, subgroup and regression analyses were conducted. RESULTS: We analysed 12 articles reporting on 24 studies involving 1817 patients and 1731 healthy controls. The pooled sensitivity was 0.77 (95% CI 0.73-0.81), the specificity was 0.81 (95% CI 0.73-0.86), the PLR was 3.99 (95% CI 2.81-5.65), the NLR was 0.28 (95% CI 0.23-0.35), the DOR was 14.18 (95% CI 8.47-23.73), and the area under the curve (AUC) was 0.85 (95% CI 0.81-0.87). Subgroup analysis revealed that multiple miRNAs can improve diagnostic performance; the pooled sensitivity of multiple miRNAs was 0.78 (95% CI 0.68-0.86), the specificity was 0.85 (95% CI 0.78-0.90), and the AUC was 0.89 (95% CI 0.86-0.91). CONCLUSION: This study suggested that circulating microRNAs may be biomarkers for early CC diagnosis.

The identification of N6-methyladenosine-related miRNAs predictive of hepatocellular carcinoma prognosis and immunotherapy efficacy.

BACKGROUND: Hepatocellular carcinoma (HCC) has a high degree of malignancy and poor prognosis. N6-methyladenosine (m6A) modifications and microRNAs (miRNAs) play pivotal roles in tumorigenesis and development. However, the role of m6A-related miRNAs in HCC has not been clarified yet. This study aimed to identify the role of m6A-miRNAs in HCC prognosis through bioinformatics analysis. METHODS: The clinicopathological information and RNA sequencing data of 369 HCC tumor tissues and 49 tumor-adjacent tissues were downloaded from the TCGA database. A total of 23 m6A regulators were extracted to evaluated the m6A-related miRNAs using Pearson's correlation analysis. Then, we selected prognosis-related m6A-miRNAs using a univariate Cox regression model and used the consensus cluster analysis to explore the characteristics of the m6A-miRNAs. The coefficient of the least absolute shrinkage and selection operator (LASSO) Cox regression was applied to construct a prognostic risk score model. The receiver operated characteristic (ROC) analysis was applied to evaluate the prognostic value of the signature. The biological functions of targeted genes were predicted by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Then, to validate the potential predictive value for prognosis, the miRNA expression profiles from the GSE76903 and GSE6857 were used. Single sample Gene Set Enrichment Analysis (ssGSEA) and Estimation of Stromal and Immune cells in Malignant Tumor tissues using Expression data (ESTIMATE) were applied to assess the immune microenvironment of HCC. Additionally, a meta-analysis was used to verify the prognostic value of the m6A-microRNAs. RT-PCR was applied to validated the expression of miRNAs in HCC tissues. Cell viability, transwell assay and RNA m6A dot blot assays of HCC cells was applied to access the function of miR-17-5p. RESULTS: The expression of 48 m6A-related miRNAs was identified and 17 prognostic m6A-miRNAs was discovered. The expression profile of those 17 miRNAs was divided into three clusters, and these clusters were associated with the tumor microenvironment (TME) and prognosis. The nine m6A-related miRNA signature was associated with the prognosis of HCC, the AUC of the ROC was 0.771(TCGA dataset), 0.788(GSE76903) and 0.646(GSE6857). The TME and the expression of immune checkpoint molecules were associated with the risk score. The meta-analysis also validated the prognostic value of the m6A-related miRNAs (miR182-5p (HR:1.58, 95%CI:1.04-2.40) and miR-17-5p (HR:1.58, 95%CI: 1.04-2.40)). The expression of miR-17-5p was upregulated in HCC tissues and miR-17-5p showed an oncogenic role in HCC cells. CONCLUSION: The clinical innovation is the use of m6A-miRNAs as biomarkers for predicting prognosis regarding immunotherapy response in HCC patients.

Identification of cell death-related biomarkers and immune infiltration in ischemic stroke between male and female patients.

Background: Stroke is the second leading cause of death and the third leading cause of disability worldwide, with ischemic stroke (IS) being the most prevalent. A substantial number of irreversible brain cell death occur in the short term, leading to impairment or death in IS. Limiting the loss of brain cells is the primary therapy target and a significant clinical issue for IS therapy. Our study aims to establish the gender specificity pattern from immune cell infiltration and four kinds of cell-death perspectives to improve IS diagnosis and therapy. Methods: Combining and standardizing two IS datasets (GSE16561 and GSE22255) from the GEO database, we used the CIBERSORT algorithm to investigate and compare the immune cell infiltration in different groups and genders. Then, ferroptosis-related differently expressed genes (FRDEGs), pyroptosis-related DEGs (PRDEGs), anoikis-related DEGs (ARDEGs), and cuproptosis-related DEGs (CRDEGs) between the IS patient group and the healthy control group were identified in men and women, respectively. Machine learning (ML) was finally used to generate the disease prediction model for cell death-related DEGs (CDRDEGs) and to screen biomarkers related to cell death involved in IS. Results: Significant changes were observed in 4 types of immune cells in male IS patients and 10 types in female IS patients compared with healthy controls. In total, 10 FRDEGs, 11 PRDEGs, 3 ARDEGs, and 1 CRDEG were present in male IS patients, while 6 FRDEGs, 16 PRDEGs, 4 ARDEGs, and 1 CRDEG existed in female IS patients. ML techniques indicated that the best diagnostic model for both male and female patients was the support vector machine (SVM) for CDRDEG genes. SVM's feature importance analysis demonstrated that SLC2A3, MMP9, C5AR1, ACSL1, and NLRP3 were the top five feature-important CDRDEGs in male IS patients. Meanwhile, the PDK4, SCL40A1, FAR1, CD163, and CD96 displayed their overwhelming influence on female IS patients. Conclusion: These findings contribute to a better knowledge of immune cell infiltration and their corresponding molecular mechanisms of cell death and offer distinct clinically relevant biological targets for IS patients of different genders.

Mechano-biomimetic hydrogel 3D cell cultivation as a strategy to improve mammalian cell protein expression.

Eukaryotic expression systems are frequently employed for the production of recombinant proteins as therapeutics as well as research tools. Among which mammalian cell protein expression approach is the most powerful one, which can express complex proteins or genetic engineered biological drugs, such as PD-1. However, the high expense, which partially derives from its low protein yielding efficiency, limited the further application of such approach in large scale production of target proteins. To address this issue, we proposed a novel technique to promote the protein production efficiency of mammal cells without using conventional genetic engineered approaches. By placing 293T cells in a hydrogel 3D cell culture platform and adjusting the stress relaxation of the matrix hydrogel, cells formed multicellular spheroids by self-organization. In particular, the multicellular spheroids have a significantly enhanced ability to transiently express multiple proteins (SHH-N, PD-1 and PDL-1). We also examined in detail the mechanism underlying this phenomenon, and found that the reorganization of cytoskeleton during spheroids formation enhances the translation process of protein by recruiting ribosomes. Overall, this finding provides a novel approach for subsequent improvement of large-scale mammalian protein expression cell systems.

MyD88 determines the protective effects of fish oil and perilla oil against metabolic disorders and inflammation in adipose tissue from mice fed a high-fat diet.

BACKGROUND: The beneficial effects of ω-3 polyunsaturated fatty acids (PUFA) vary between different sources. However, there is a paucity of comparative studies regarding the effects and mechanisms of marine and plant ω-3 PUFA on obesity. OBJECTIVE: The aim of this study was to evaluate the effects of fish oil (FO) and perilla oil (PO) on glucolipid metabolism, inflammation, and adipokine in mice fed a high-fat (HF) diet in association with the contribution of toll-like receptor 4 (TLR4)/myeloid differentiation primary response 88 (MyD88) pathway. METHODS: C57BL/6J mice and MyD88-/- mice were randomly divided into 4 groups: normal chow diet, HF diet, HF diet accompanied by daily gavage with either FO or PO. After 4 weeks, blood biochemistries, adipocyte histology, mRNA, and protein expression of MyD88-dependent and -independent pathways of TLR4 signaling in epididymal adipose tissue were measured. RESULTS: In C57BL/6J mice, there were no statistical differences between FO and PO in decreasing body weight, glucose, insulin, triglyceride, total cholesterol, interleukin-6, and increasing adipocyte counts. FO and PO decreased mRNA and protein expression of TLR4, MyD88, tumor necrosis factor receptor-associated factor 6, inhibitor of nuclear factor kappa B kinase beta and nuclear factor-kappa B p65. In MyD88-/- mice, the beneficial effects of FO and PO on HF diet-induced metabolism abnormalities and inflammation were abolished. FO and PO had no impacts on mRNA and protein expression of receptor-interacting protein-1, interferon regulate factor 3, and nuclear factor-kappa B p65. CONCLUSION: FO and PO exhibit similar protective effects on metabolic disorders and inflammation through inhibiting TLR4 signaling in a manner dependent on MyD88. These findings highlight plant ω-3 PUFA as an attractive alternative source of marine ω-3 PUFA and reveal a mechanistic insight for preventive benefits of ω-3 PUFA in obesity and related metabolic diseases.

MicroRNA-224 modulates chemosensitivity of breast cancer cells to docetaxel by apoptosis inhibitor 5.

PURPOSE: The view that microRNA-224 (miR-224) may lead to tumorigenesis has been accepted in many studies. However, its role remains unclear in modulating the chemosensitivity of breast cancer cells to docetaxel (DOC). Thus, the aim of this study was to estimate what's the role of miR-224 in the chemosensitivity of breast cancer cells to DOC. METHODS: The role of miR-224 in breast cancer cells was analyzed using CCK-8 assay, real-time PCR, flow cytometry assay and Western blot. Dual-luciferase reporter assay and API-5-siRNA technology were performed to analyze the association between miR-224 and Apoptosis inhibitor 5 (API-5). RESULTS: Overexpression of miR-224 could significantly decrease the chemosensitivity of MCF-7 breast cancer cells to DOC. The luciferase activity of MCF-7/DOC cells containing wild-type 3'UTR of API-5 could be inhibited by miR-224 mimics. Similarly, the chemoresistance of MCF-7 cells to DOC induced by miR-224 mimics could be partially reversed by API-5-siRNA. CONCLUSION: An inverse association between miR-224 and API-5 in breast cancer cells was revealed. Dysregulation of miR-224 plays a vital role in the acquired DOC resistance of breast cancer and at least partially via targeting API-5.

Different effects of high-fat diets rich in different oils on lipids metabolism, oxidative stress and gut microbiota.

The study aimed to investigate the different effects of high-fat (HF) diets rich in different oils on lipid metabolism, oxidative stress, and gut mirobiota. C57BL/6 mice were divided into 4 groups: (1) control group (CG) was fed with normal diet, (2) olive oil (OO) group was fed with high-fat diet containing OO, (3) lard oil (LO) group was fed with high-fat diet containing LO, (4) soybean oil (SO) group was fed with high-fat diet containing SO. After 12 weeks, serum lipids, and oxidative stress indices were analyzed. Gut microbiota analysis was carried out based on the sequencing results of 16S rRNA. High fat diet can increase serum and liver lipids and upregulate sterol regulatory element-binding protein-1c related genes expression. Serum and liver malondialdehyde (MDA) levels in LO group were significantly higher than those in CG and OO groups. In CG, the family Muribaculaceae, Lactobacillaceae, Lachnospiraceae and Desulfovibrionaceae had the large effect sizes. HF diets resulted in the increase of Actinobacteria and Enterococcaceae abundance, and the decrease of Bacteroidetes, Proteobacteria Lactobacillales and microbiota diversity. The abundance of Actinobacteria and Lactobacillales is the link to the serum TC and MDA levels. HF diets have the harmful influence on the serum lipids, oxidative stress and endothelial function. They can also cause gut microbiota dysbiosis.

The validation of a questionnaire to delineate clinical research coordinator roles in China.

PURPOSE: The purpose of this study was to evaluate the roles of the clinical research coordinator (CRC) in clinical oncology trials. DESIGN AND METHODS: An E-questionnaire that includes 10 sections with 155 items total and is based on the Clinical Trials Nursing Questionnaire (CTNQ) was designed to determine the conditions of demographics, competences, activities, and psychology for Chinese CRCs. Eighty-two CRCs from three different provinces in China were invited to join this study anonymously. Cronbach's α and split-half reliability were calculated to assess the reliability and validity of the questionnaire. Additionally, the Wilcoxon rank-sum test was used to find the similarity and difference between the importance of the roles of CRCs and their frequency. The STROBE checklist for observational research has been following for presenting the research (see File S1). FINDINGS: Cronbach's α values of the Chinese version of the questionnaire for the frequency scale and the importance scale were .965 and .961, respectively. The split-half reliability coefficients were 0.866 and 0.805, respectively. Pearson correlation coefficients of the subscales indicated that the correlation between each item and its dimension was greater than its correlation with the other components (P < .05). Exploratory factor analysis results show that three common factors were extracted by principal component analysis and had eigenvalues greater than 1 and that the cumulative contribution rate was 69.415%. PRACTICE IMPLICATIONS: The Chinese version of the questionnaire has good reliability and validity for CRCs in China, which could be promoted in evaluating clinical research coordinator roles in China.

Perilla Oil Supplementation Improves Hypertriglyceridemia and Gut Dysbiosis in Diabetic KKAy Mice.

SCOPE: The aim of this study is to examine whether perilla oil supplementation improves glucolipid metabolism and modulates gut microbiota in diabetic KKAy mice. METHODS AND RESULTS: The successfully established diabetic KKAy mice are randomized into four groups: diabetic model (DM), low-dose perilla oil (LPO), middle-dose perilla oil (MPO), and high-dose perilla oil (HPO). C57BL/6J mice are fed a chow diet as normal control (NC). At the end of 12 weeks, mice are euthanized and glucolipid indications are analyzed. Gut microbiota analysis is carried out based on the sequencing results on V4 region of 16S rRNA. Although serum glucose, insulin, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, abundance-based coverage estimator, and shannon are unchanged, serum triglyceride significantly decreases in LPO compared with DM. The histopathological changes of hepatocellular macrovesicular steatosis and adipocyte hypertrophy are ameliorated by perilla oil supplementation. Blautia is significantly decreased in LPO, MPO, and HPO, compared with DM. Nonmetric multidimensional scaling analysis shows NC and LPO are relatively coherent. CONCLUSION: These findings indicate that dietary supplementation with perilla oil can improve hypertriglyceridemia and gut dysbiosis in diabetic KKAy mice, which can be associated with potential benefits to human health.

Effects of palm olein and olive oil on serum lipids in a Chinese population: a randomized, double-blind, cross-over trial.

BACKGROUND AND OBJECTIVES: As the most widely produced edible vegetable oil, palm oil is known as to contain a high level of saturated fatty acid, which was thought to adversely affect serum lipid profiles. However, recent studies have shown no influence or benefits of palm oil on serum lipids. The potential nutritional value of palm oil is attributed to the high mono-unsaturation at the crucial sn2-position of the oil's triacylglycerols, as with the so-called 'healthy' olive oil (OO). The aim of this study was to further test this hypothesis and evaluate the effects of consuming palm olein versus olive oil on serum lipid profiles in a Chinese population. METHODS AND STUDY DESIGN: In total, 120 participants were recruited from a spinnery in Yixing city and randomly divided into two groups (palm olein or olive oil) to conduct a 2×2 crossover trial for 2 months' intervention with 2-week washout periods. Each participant was provided 48 g of test oil per day. At the end of each period, anthropometry, and blood lipid indices were measured to determine the effects of palm olein and olive oil. RESULTS: Palm olein and olive oil consumption had no significantly different effect on BMI, on serum total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triacylglycerol (TG), Apo B, fasting glucose, or insulin concentrations (all p>0.05). CONCLUSIONS: In a dietary crossover trial, palm olein and olive oil had no recognisably different effects on body fatness or blood lipids in a healthy Chinese population.

An investigation report about the lack of VB1.

From August 1 to September 26, 2012, 30 workers who had the symptom of edema in both lower limbs were reported in Yancheng, Four severe edemas in both lower limbs workers were sent to the hospital, and one person of the workers died. The epidemiology investigation and laboratory testing (in the urine loading tests, the concentration of VB1 of the workers living in the factory for more than half of the year decreased far below the normal values) were conducted to explore the cause. Furthermore, after one-week intramuscular injection of thiamine treatment, the clinical symptoms of the workers had been improved greatly. Finally, the lake of the thiamine (VB1) was considered to be the reason of this incident.

[Determination of total Bromine in urine by inductively coupled plasma mass spectrometry (ICP-MS)].

OBJECTIVE: To establish a method to determine total bromine in urine. METHOD: Diluted urine samples were directly introduced into ICP-MS then quantized by standard curve. RESULT: Total bromine in urine was linear within 1.0~50 mg/L with r > 0.999, When spiked at a concentration of 0.020 mg/L, 0.050 mg/L, 0.150 mg/L, the recovery was 95%~98%, intra-assay precision was 1.4% 3.2%, inter-assay precision was 3.4% to 5.0%. Urine could store in -20 °C refrigerator 3 months without any bromine loss. CONCLUSION: Using ICP-MS to determine the urinary total bromine, the method is fast, accurate, wide linear range of features, could meet with the requirement of Part 5 of occupational health standards guide: Method determination of chemical substances in biological materials (GBZ/T 210.5-2008), a strong competitive advantage in a wide range of survey, suitable for promotion.