The Expression and Clinical Value of miR-221 and miR-320 in the Plasma of Women with Gestational Diabetes Mellitus.

BACKGROUND: The goal was to investigate the expression of plasma miR-221 and miR-320 in gestational diabetes mellitus (GDM) and to further explore the relationship between miRNA and risk factors for GDM. METHODS: This study included 85 GDM and 85 age-matched normal pregnant women who visited our hospital from January 2019 to January 2020. Real-time polymerase chain reaction (RT-qPCR) was used to determine the expression of miR-221 and miR-320 in the plasma of pregnant women. The correlation analysis was used to detect the relationship between miR-221, miR-320, and risk factors of GDM, including homeostatic model assessment for insulin resistance (HOMA-IR), percentage of glycosylated hemoglobin (HbA1c), and the prepregnancy BMI. The receiver operating characteristic curve (ROC) was used to determine the diagnostic value of miR-320 and miR-221 in GDM. RESULTS: Compared with normal pregnant women, the expression of miR-221 and miR-320 in GDM was significantly higher (p < 0.05). The results also demonstrate that the expression of miR-221 and miR-320 increases grad-ually with the development of pregnancy in GDM at 24 weeks, 28 weeks, and 32 weeks (p < 0.05). Spearman's correlation analysis confirmed that the expression level of miR-221 and miR-320 in the plasma of GDM is positively correlated with the HOMA-IR and HbA1c, but has no significant correlation with pre-pregnancy BMI. The area under the curve (AUC) values of miR-221 and miR-320 were 0.862 and 0.853, respectively. Meanwhile, the area under the combined detection curve is 0.904. CONCLUSIONS: Plasma miR-221 and miR-320 are significantly elevated in GDM, and are positively correlated with HbA1c and HOMA-IR. The high expression of miR-221 and miR-320 in the peripheral plasma of pregnant women may directly or indirectly participate in the occurrence and development of GDM or may become a new target for the diagnosis, treatment, and prognosis of GDM.

The differential demographic pattern of coronavirus disease 2019 fatality outside Hubei and from six hospitals in Hubei, China: a descriptive analysis.

BACKGROUND: The coronavirus disease 2019 (COVID-19) epidemic has been largely controlled in China, to the point where case fatality rate (CFR) data can be comprehensively evaluated. METHODS: Data on confirmed patients, with a final outcome reported as of 29 March 2020, were obtained from official websites and other internet sources. The hospitalized CFR (HCFR) was estimated, epidemiological features described, and risk factors for a fatal outcome identified. RESULTS: The overall HCFR in China was estimated to be 4.6% (95% CI 4.5-4.8%, P < 0.001). It increased with age and was higher in males than females. Although the highest HCFR observed was in male patients ≥70 years old, the relative risks for death outcome by sex varied across age groups, and the greatest HCFR risk ratio for males vs. females was shown in the age group of 50-60 years, higher than age groups of 60-70 and ≥ 70 years. Differential age/sex HCFR patterns across geographical regions were found: the age effect on HCFR was greater in other provinces outside Hubei than in Wuhan. An effect of longer interval from symptom onset to admission was only observed outside Hubei, not in Wuhan. By performing multivariate analysis and survival analysis, the higher HCFR was associated with older age (both P < 0.001), and male sex (both P < 0.001). Only in regions outside Hubei, longer interval from symptom onset to admission, were associated with higher HCFR. CONCLUSIONS: This up-to-date and comprehensive picture of COVID-19 HCFR and its drivers will help healthcare givers target limited medical resources to patients with high risk of fatality.

Detection of diluted contaminants on chicken carcasses using a two-dimensional scatter plot based on a two-dimensional hyperspectral correlation spectrum.

A two-dimensional (2D) scatter plot method based on the 2D hyperspectral correlation spectrum is proposed to detect diluted blood, bile, and feces from the cecum and duodenum on chicken carcasses. First, from the collected hyperspectral data, a set of uncontaminated regions of interest (ROIs) and four sets of contaminated ROIs were selected, whose average spectra were treated as the original spectrum and influenced spectra, respectively. Then, the difference spectra were obtained and used to conduct correlation analysis, from which the 2D hyperspectral correlation spectrum was constructed using the analogy method of 2D IR correlation spectroscopy. Two maximum auto-peaks and a pair of cross peaks appeared at 656 and 474 nm. Therefore, 656 and 474 nm were selected as the characteristic bands because they were most sensitive to the spectral change induced by the contaminants. The 2D scatter plots of the contaminants, clean skin, and background in the 474- and 656-nm space were used to distinguish the contaminants from the clean skin and background. The threshold values of the 474- and 656-nm bands were determined by receiver operating characteristic (ROC) analysis. According to the ROC results, a pixel whose relative reflectance at 656 nm was greater than 0.5 and relative reflectance at 474 nm was lower than 0.3 was judged as a contaminated pixel. A region with more than 50 pixels identified was marked in the detection graph. This detection method achieved a recognition rate of up to 95.03% at the region level and 31.84% at the pixel level. The false-positive rate was only 0.82% at the pixel level. The results of this study confirm that the 2D scatter plot method based on the 2D hyperspectral correlation spectrum is an effective method for detecting diluted contaminants on chicken carcasses.

[Study on the Fast Quantitative Analysis of the Content of DME Adulterated in LPG].

A new analytical method of analyzing dimethyl ether (DME) content in liquefied petroleum gas (LPG) is proposed in this paper. An unsolved problem about quick detection of the composition of LPG has been settled with this method. A set of precise preparation apparatus for DME/ LPG solution and a set of quickly analytical system of LPG based on near infrared technology were designed. The analytical equipment can be conveniently connected to the sampling cylinder because it can bear 3.5 MPa pressure. Oblique projection algorithm was used to separate the pure spectra of DME from that of the LPG's solutions. The standard curve of the concentration of DME (c) has been built by using the Intensity (I) of pure signal of DME in the LPG solution and the concentrations. The correlation coefficient of the equation is 0.999 4. The result of external validation shows that the relative error is less than 2.0%. The new method has the advantages such as fast, easy and noneed of expensive multivariate modeling.

Synergistic effects of the bimetallic Ni-Fe systems and their application in the reductive amination of polyether polyols.

We report the synthesis of xNi-yFe/γ-Al2O3 catalysts which were applied to the reductive amination of polypropylene glycol (PPG) for the preparation of polyether amine (PEA). The catalysts were characterized by N2-sorption, X-ray diffraction, H2-temperature programmed reduction, energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy to reveal the synergistic effect of the bimetallic Ni-Fe-loaded catalysts. It was found that in the reductive amination of PPG to PEA, the conversion and product selectivity of the reaction were closely related to the types of active centers of the catalyst. In particular, the surface Ni0 content increased by adding Fe as a promoter, with a maximum Ni0 content on the 15Ni-7.5Fe/Al2O3 catalyst, which also led to the highest conversion rate (>99%). In addition, no deactivation was observed after three cycles of reaction carried out by the catalyst.

Inhibition of JNK Signaling Overcomes Cancer-Associated Fibroblast-Mediated Immunosuppression and Enhances the Efficacy of Immunotherapy in Bladder Cancer.

Currently, only 20-40% of cancer patients benefit from immune checkpoint inhibitors. Understanding the mechanisms underlying the immunosuppressive tumor microenvironment (TME) and characterizing dynamic changes in the immunological landscape during treatment are critical for improving responsiveness to immunotherapy. Here, we identified JNK signaling in cancer-associated fibroblasts (CAFs) as a regulator of the immunosuppressive tumor microenvironment. Single-cell RNA sequencing of bladder cancer treated with a JNK inhibitor revealed enhanced cytotoxicity and effector functions of CD8+ T cells. In untreated tumors, CAFs interacted frequently with CD8+ T cells and mediated their exhaustion. JNK inhibition abrogated the immunosuppression function of CAFs by downregulating the expression of TSLP, thereby restoring CD8+ T cell cytotoxicity. In addition, blockade of CAF-derived TSLP in combination with anti-PD1 treatment promoted tumor elimination by CD8+ T cells in vivo. Collectively, these results indicate that JNK signaling plays an important immunosuppressive role in the tumor microenvironment by promoting expression of TSLP in CAFs and suggest that inhibiting JNK signaling could be a promising immunotherapeutic strategy for cancer treatment.

Combined Inhibition of PI3K and STAT3 signaling effectively inhibits bladder cancer growth.

Bladder cancer is characterized by aberrant activation of the phosphatidylinositol-3-OH kinase (PI3K) signaling, underscoring the significance of directing therapeutic efforts toward the PI3K pathway as a promising strategy. In this study, we discovered that PI3K serves as a potent therapeutic target for bladder cancer through a high-throughput screening of inhibitory molecules. The PI3K inhibitor demonstrated a robust anti-tumor efficacy, validated both in vitro and in vivo settings. Nevertheless, the feedback activation of JAK1-STAT3 signaling reinstated cell and organoid survival, leading to resistance against the PI3K inhibitor. Mechanistically, the PI3K inhibitor suppresses PTPN11 expression, a negative regulator of the JAK-STAT pathway, thereby activating STAT3. Conversely, restoration of PTPN11 enhances the sensitivity of cancer cells to the PI3K inhibitor. Simultaneous inhibition of both PI3K and STAT3 with small-molecule inhibitors resulted in sustained tumor regression in patient-derived bladder cancer xenografts. These findings advocate for a combinational therapeutic approach targeting both PI3K and STAT3 pathways to achieve enduring cancer eradication in vitro and in vivo, underscoring their promising therapeutic efficacy for treating bladder cancer.

The Mobility of Eurasian Avian-like M2 Is Determined by Residue E79 Which Is Essential for Pathogenicity of 2009 Pandemic H1N1 Influenza Virus in Mice.

In 2009, a novel H1N1 influenza virus caused the first influenza pandemic of the 21st century. Studies have shown that the influenza M gene played important roles in the pathogenicity and transmissibility of the 2009 H1N1 pandemic ((H1N1)pdm09), whilst the underlying mechanism remains unclear. The influenza M gene encodes two proteins, matrix protein 1 and matrix protein 2, which play important roles in viral replication and assembly. In this study, it is found that the M2 protein of the (H1N1)pdm09 virus showed a lower mobility rate than the North America triple-reassortant influenza M2 protein in Polyacrylamide Gel Electrophoresis (PAGE). The site-directed mutations of the amino acids of (H1N1)pdm09 M2 revealed that E79 is responsible for the mobility rate change. Further animal studies showed that the (H1N1)pdm09 containing a single M2-E79K was significantly attenuated compared with the wild-type virus in mice and induced lower proinflammatory cytokines and IFNs in mouse lungs. Further in vitro studies indicated that this mutation also affected NLRP3 inflammasome activation. To reveal the reason why they have different mobility rates, a circular dichroism spectra assay was employed and showed that the two M2 proteins displayed different secondary structures. Overall, our findings suggest that M2 E79 is important for the virus replication and pathogenicity of (H1N1)pdm09 through NLRP3 inflammasome and proinflammatory response.

Safety Risk Estimation of Construction Project Based on Energy Transfer Model and System Dynamics: A Case Study of Collapse Accident in China.

Analyzing and understanding the occurrence and evolution mechanisms of construction accidents are important for construction safety management. This study proposed a hybrid approach of integrating the energy transfer model (ETM) and system dynamics (SD) theory to delineate the entire evolution stage of the construction accident. Specifically, the Fengcheng Power Plant construction platform collapse accident (FPCA) was taken as a practical case study. First, the ETM is applied to demonstrate the evolving nature of the accident. Then, the network of the accident-causing factors is constructed using the SD theory to analyze the dynamic change characteristics. The results indicate that the accident was caused by risk factors with complex interactions at the management level. An energy constraint failure occurred when the transfer of dangerous energy transpired at the physical entity level, inducing the event. The proposed approach can provide a useful reference for safety risk estimation and management in future major construction projects.

JNK Signaling Promotes Bladder Cancer Immune Escape by Regulating METTL3-Mediated m6A Modification of PD-L1 mRNA.

The RNA N6-methyladenosine (m6A) writer methyltransferase-like 3 (METTL3) is upregulated in many types of cancer and promotes cancer progression by increasing expression of several oncogenes. Therefore, a better understanding of the mechanisms regulating METTL3 expression and the key targets of METTL3 in cancer cells could provide new therapeutic targets. In this study, we found that activated JNK signaling is associated with increased METTL3 expression in bladder cancer. Knockdown of JNK1 or administration of a JNK inhibitor impaired the binding of c-Jun with the METTL3 promoter, thereby decreasing the expression of METTL3 and global RNA m6A levels. Moreover, RNA m6A sequencing indicated enrichment of m6A in the 3'-UTR of immune checkpoint PD-L1 mRNA, which could be recognized by the m6A reader IGF2BP1 to mediate RNA stability and expression levels of PD-L1. Inhibition of JNK signaling suppressed m6A abundance in PD-L1 mRNA, leading to decreased PD-L1 expression. Functionally, METTL3 was essential for bladder cancer cells to resist the cytotoxicity of CD8+ T cells by regulating PD-L1 expression. Additionally, JNK signaling contributed to tumor immune escape in a METTL3-dependent manner both in vitro and in vivo. These data reveal the JNK/METTL3 axis as a mechanism of aberrant m6A modification and immune regulation in bladder cancer. SIGNIFICANCE: The identification of a novel m6A-dependent mechanism underlying immune system evasion by bladder cancer cells reveals JNK signaling as a potential target for bladder cancer immunotherapy.

Pre-exposure to Streptococcus suis improved survival of influenza virus co-infection in mice.

The synergism of the influenza virus and respiratory tract pathogens is known to exacerbate diseases in both humans and animals. The mechanism of the co-infection of associated respiratory tract pathogens is explored in this study. Co-infection has a directional effect when influenza virus or other pathogens occur in a different order. In the present study, we used a mouse animal model to study the synergism of influenza virus and Streptococcus suis co-infection in different orders of administration. We found that the group infected with bacteria alone did not show any clinical symptoms, but the group infected with the virus alone showed 100 % mortality and clinical signs typical in infected mice. In the bacteria infected following virus pre-exposure group, the mice died before the virus-infected group and showed severer clinical signs. When the influenza virus was administered after the bacteria, the infected mice showed reduced mortality compared with mice administered the influenza virus alone. The results indicated that the order of infection significantly affected the outcome of the co-infection of these two pathogens in the mice. However, the underlying mechanism was unclear. Therefore, a transcriptome analysis of mouse lungs was conducted to explore the potential mechanism. The results showed that inflammation and cell damage signaling pathways were upregulated, which may have contributed to the increased mortality in the secondary bacterial infection group. Upregulated innate immunity may have been a major cause of reduced mortality when the bacteria were inoculated before the virus infection.

Eurasian Avian-like M1 Plays More Important Role than M2 in Pathogenicity of 2009 Pandemic H1N1 Influenza Virus in Mice.

Reassortant variant viruses generated between 2009 H1N1 pandemic influenza virus [A(H1N1)pdm09] and endemic swine influenza viruses posed a potential risk to humans. Surprisingly, genetic analysis showed that almost all of these variant viruses contained the M segment from A(H1N1)pdm09, which originated from Eurasian avian-like swine influenza viruses. Studies have shown that the A(H1N1)pdm09 M gene is critical for the transmissibility and pathogenicity of the variant viruses. However, the M gene encodes two proteins, M1 and M2, and which of those plays a more important role in virus pathogenicity remains unknown. In this study, the M1 and M2 genes of A(H1N1)pdm09 were replaced with those of endemic H3N2 swine influenza virus, respectively. The chimeric viruses were rescued and evaluated in vitro and in mice. Both M1 and M2 of H3N2 affected the virus replication in vitro. In mice, the introduction of H3N2 M1 attenuated the chimeric virus, where all the mice survived from the infection, compared with the wild type virus that caused 100 % mortality. However, the chimeric virus containing H3N2 M2 was still virulent to mice, and caused 16.6% mortality, as well as similar body weight loss to the wild type virus infected group. Compared with the wild type virus, the chimeric virus containing H3N2 M1 induced lower levels of inflammatory cytokines and higher levels of anti-inflammatory cytokines, whereas the chimeric virus containing H3N2 M2 induced substantial pro-inflammatory responses, but higher levels of anti-inflammatory cytokines. The study demonstrated that Eurasian avian-like M1 played a more important role than M2 in the pathogenicity of A(H1N1)pdm09 in mice.

The m6A Methylation-Regulated AFF4 Promotes Self-Renewal of Bladder Cancer Stem Cells.

The dynamic N6-methyladenosine (m6A) modification of mRNA plays a role in regulating gene expression and determining cell fate. However, the functions of m6A mRNA modification in bladder cancer stem cells (BCSCs) have not been described. Here, we show that global RNA m6A abundance and the expression of m6A-forming enzyme METTL3 are higher in BCSCs than those in non-CSCs of bladder cancer (BCa) cells. The depletion of the METTL3 inhibited the self-renewal of BCSCs, as evidenced by decreased ALDH activity and sphere-forming ability. Mechanistically, METTL3 regulates the m6A modification and thereby the expression of AF4/FMR2 family member 4 (AFF4), knockdown of which phenocopies the METTL3 ablation and diminishes the tumor-initiating capability of BCSCs in vivo. AFF4 binds to the promoter regions and sustains the transcription of SOX2 and MYC which have critical biological functions in BCSCs. Collectively, our results demonstrate the critical roles of m6A modification in self-renewal and tumorigenicity of BCSCs through a novel signaling axis of METTL3-AFF4-SOX2/MYC.

The m6A methyltransferase METTL3 promotes bladder cancer progression via AFF4/NF-κB/MYC signaling network.

N6-methyladenosine (m6A) is the most abundant modification in eukaryotic messenger RNAs (mRNAs), and plays important roles in many bioprocesses. However, its functions in bladder cancer (BCa) remain elusive. Here, we discovered that methyltransferase-like 3 (METTL3), a major RNA N6-adenosine methyltransferase, was significantly up-regulated in human BCa. Knockdown of METTL3 drastically reduced BCa cell proliferation, invasion, and survival in vitro and tumorigenicity in vivo. On the other hand, overexpression of METTL3 significantly promoted BCa cell growth and invasion. Through transcriptome sequencing, m6A sequencing and m6A methylated RNA immuno-precipitation quantitative reverse-transcription polymerase chain reaction, we revealed the profile of METTL3-mediated m6A modification in BCa cells for the first time. AF4/FMR2 family member 4 (AFF4), two key regulators of NF-κB pathway (IKBKB and RELA) and MYC were further identified as direct targets of METTL3-mediated m6A modification. In addition, we showed that besides NF-κB, AFF4 binds to the promoter of MYC and promotes its expression, implying a novel multilevel regulatory network downstream of METTL3. Our results uncovered an AFF4/NF-κB/MYC signaling network operated by METTL3-mediated m6A modification and provided insight into the mechanisms of BCa progression.

Low doses of decitabine improve the chemotherapy efficacy against basal-like bladder cancer by targeting cancer stem cells.

Low dose treatment with the DNA methylation inhibitor decitabine has been shown to be applicable for the management of certain types of cancer. However, its antitumor effect and mechanisms are context dependent and its activity has never been systematically studied in bladder cancer treatment. We used mouse models, cultured cell lines and patient-derived xenografts to demonstrate that low dose decitabine treatment remarkably enhanced the effects of cisplatin and gemcitabine on basal-like bladder cancer both in vivo and in vitro. Genetic lineage tracing revealed that the stemness of a bladder cancer stem cell population was inhibited by decitabine treatment in mice. These effects were accompanied by decreases in genome-wide DNA methylation, gene re-expression, and changes in key cellular regulatory pathways such as STAT3 signaling. These results indicate that this DNA-demethylating reagent is a promising therapeutic approach for basal-like bladder cancer treatment.

SOX2 Is a Marker for Stem-like Tumor Cells in Bladder Cancer.

It has been reported that functionally distinct cancer stem cells (CSCs) exist in human bladder cancer (BCa). Here, we found that Sox2, a transcription factor that is well characterized as a marker for stem cells, is upregulated in both mouse and human BCa. Sox2 expression is absent in normal urothelial cells, but it begins to be expressed in pre-neoplastic bladder tumors and continues to be expressed in invasive mouse BCa. Using s as a reporter of Sox2 transcriptional expression, we demonstrated that Sox2-expressing cells mark a subpopulation of tumor cells that fuel the growth of established BCa. SOX2-positive cells also expressed other previously reported BCa CSC markers, including Keratin14 (KRT14) and CD44v6. Ablation of Sox2-expressing cells within primary invasive BCa led to enhanced tumor regression, supporting the essential role of SOX2-positive cells in regulating BCa maintenance and progression. Our data show that Sox2 is a marker of bladder CSCs and indicate it as a potential clinical target for BCa therapy.

High expression of microRNA-183/182/96 cluster as a prognostic biomarker for breast cancer.

More sensitive and effective diagnostic markers for the detection of breast cancer are urgently needed. The microRNA-183/182/96 cluster has been reported to be involved in tumorigenesis and progression in a variety of cancers, and it is a promising cancer prognostic biomarker. The goal of this study was to determine the expression levels of the miR-183/182/96 cluster in breast cancer tissues and evaluate its prognostic role in breast cancer. Real-time quantitative polymerase chain reaction analysis (qRT-PCR) was used to detect the expression levels of the miR-183/182/96 cluster in 41 breast cancer tissues and adjacent normal tissues (control tissues) and also in different mammary cell lines. In situ hybridization (ISH) of the miR-183/182/96 cluster on 131 tissue microarrays (TMAs) was used to statistically analyze its prognostic role. The miR-183/182/96 cluster levels were significantly higher in breast cancer tissues than in control tissues. The miR-183/182/96 cluster was also upregulated in human breast cancer cell lines. An increased miR-183/182/96 cluster level was correlated with local relapse, distant metastasis and poor clinical outcomes. Our findings improve our understanding of the expression level of the miR-183/182/96 cluster in breast cancer and clarify the role of the miR-183/182/96 cluster as a novel prognostic biomarker for breast cancer.

Cytokeratin19-2g2, a novel fragment of cytokeratin19 in serum, indicating a more invasive behavior and worse prognosis in breast cancer patients.

BACKGROUND: Various studies have been searching for new tumor biomarkers for breast cancer for years. However, so far, few markers have been proved clinically useful except CA153. Based on knowledge that most adenocarcinomas including breast carcinoma expressed Cytokeratin19, the authors studied CK19-2G2,a novel fragment of cytokeratin19 shedding into serum in breast cancer patients. PATIENTS AND METHODS: The serum samples of four hundred and seventeen patients including three hundred and three (fifty-four DCIS and two hundred and forty-nine stage I-III) PBC patients and one hundred and fourteen MBC patients, eighty-one healthy controls and twenty-one breast benign disease patients were provided for measurement of CK19-2G2, CEA and CA153.The correlation between clinicopathological characters, prognosis and CK19-2G2 levels was further studied. RESULTS: The serum CK19-2G2 levels in breast cancer patients were significantly higher than that in healthy and benign controls. For breast cancer patients, CK19-2G2 levels in MBC were significantly higher than that in PBC patients. The sensitivities of CK19-2G2 for breast carcinoma are as high as CEA and CA153, and up to 71% in MBC patients. Serum CK19-2G2 levels (≥2 mU/mL) were associated with pathological stages, tumor size (≥2 cm), lymph node involvement, and HER2 status. Multivariate analysis revealed that high serum CK19-2G2 level was an independent factor for relapse (P = 0.029) and death (P = 0.040) in breast cancer patients. CONCLUSION: Serum CK19-2G2 may be an independent indicator for prognosis and a candidate marker for monitoring metastasis in breast cancer.

[Determination of nitrogen compounds in catalytic gasolines by gas chromatography-nitrogen chemiluminescence detection].

A method for the separation and determination of nitrogen compounds in catalytic gasolines by gas chromatography-nitrogen chemiluminescence detection (GC-NCD) was established. The effects of the flow rate of carrier gas and the oven temperature on the resolution were studied. More than 20 nitrogen compounds, including amines, pyridine, aniline, 2-methyl aniline, 3-methyl aniline, 4-methyl aniline, quinoline, and indole, in catalytic gasoline were identified based on the retention time of some pure nitrogen compounds and by gas chromatography-mass spectrometry. The relative standard deviations of the peak areas of main nitrogen compounds in a catalytic gasoline sample were less than 2.5% and the detection limits for nitrogen were 1.0 mg/L under the chosen conditions. The linear ranges were 1.0-100 mg/L nitrogen for each nitrogen compound. The correlation coefficients were more than 0.998. The method can be successfully applied for the determination of each nitrogen compound in different catalytic gasolines.