Forkhead-associated phosphopeptide binding domain 1 (FHAD1) deficiency impaired murine sperm motility.

Background: Genetic knockout-based studies conducted in mice provide a powerful means of assessing the significance of a gene for fertility. Forkhead-associated phosphopeptide binding domain 1 (FHAD1) contains a conserved FHA domain, that is present in many proteins with phospho-threonine reader activity. How FHAD1 functions in male fertility, however, remains uncertain. Methods: Fhad1-/- mice were generated by CRISPR/Cas9-mediated knockout, after which qPCR was used to evaluate changes in gene expression, with subsequent analyses of spermatogenesis and fertility. The testis phenotypes were also examined using immunofluorescence and histological staining, while sperm concentrations and motility were quantified via computer-aided sperm analysis. Cellular apoptosis was assessed using a TUNEL staining assay. Results: The Fhad1-/-mice did not exhibit any abnormal changes in fertility or testicular morphology compared to wild-type littermates. Histological analyses confirmed that the testicular morphology of both Fhad1-/-and Fhad1+/+ mice was normal, with both exhibiting intact seminiferous tubules. Relative to Fhad1+/+ mice, however, Fhad1-/-did exhibit reductions in the total and progressive motility of epididymal sperm. Analyses of meiotic division in Fhad1-/-mice also revealed higher levels of apoptotic death during the first wave of spermatogenesis. Discussion: The findings suggest that FHAD1 is involved in both meiosis and the modulation of sperm motility.

The Role of Metabolomics and Microbiology in Urinary Tract Infection.

One of the common illnesses that affect women's physical and mental health is urinary tract infection (UTI). The disappointing results of empirical anti-infective treatment and the lengthy time required for urine bacterial culture are two issues. Antibiotic misuse is common, especially in females who experience recurrent UTI (rUTI). This leads to a higher prevalence of antibiotic resistance in the microorganisms that cause the infection. Antibiotic therapy will face major challenges in the future, prompting clinicians to update their practices. New testing techniques are making the potential association between the urogenital microbiota and UTIs increasingly apparent. Monitoring changes in female urinary tract (UT) microbiota, as well as metabolites, may be useful in exploring newer preventive treatments for UTIs. This review focuses on advances in urogenital microbiology and organismal metabolites relevant to the identification and handling of UTIs in an attempt to provide novel methods for the identification and management of infections of the UT. Particular attention is paid to the microbiota and metabolites in the patient's urine in relation to their role in supporting host health.

Preparation of Budesonide-Loaded Liposomal Nanoparticles for Pulmonary Delivery and Their Therapeutic Effect in OVA-Induced Asthma in Mice.

Purpose: Inhaled corticosteroids, including budesonide (BUD), are widely employed for the treatment of asthma. However, the frequent use of corticosteroids is associated with numerous adverse effects and poses challenges to ongoing drug therapy and patient adherence. Budesonide liposomal nanoparticles (BUD-LNPs) were developed to improve the bioavailability of the drug and thereby improve the effectiveness of asthma treatment. Methods: BUD-LNPs were prepared via thin-film hydration, and the characterizations, stability, and in vitro release of BUD-LNPs were studied. In vitro cellular uptake was observed by laser-scanning confocal microscope (LSCM) and flow cytometry. And the in vitro anti-inflammatory activity of BUD-LNPs was evaluated by measuring the expression of pro-inflammatory cytokines in activated macrophages. Besides, the accumulation time in the lung of drugs delivered via liposomal carriers and free drugs was compared in vivo. And the in vivo therapeutic efficacy of BUD-LNPs was assessed in OVA-induced asthmatic mice. Finally, in vivo biosafety assessment was performed. Results: The particle size, PDI, and zeta potential of BUD-LNPs were 127.63±1.33 nm, 0.27±0.02, and 3.33±0.13 mV, respectively. BUD-LNPs exhibited excellent biosafety and anti-inflammatory activity in vitro. Furthermore, compared with the free drugs, the utilization of liposomal nano-vehicles for drugs delivery could effectively extend the duration of drugs accumulation in the pulmonary system. Additionally, treatment with BUD-LNPs alleviated airway hyperresponsiveness, reduced airway mucus secretion, and mitigated pulmonary inflammation in OVA-induced asthmatic mice. And the BUD-LNPs demonstrated superior therapeutic efficacy compared to free BUD. Conclusion: BUD-LNPs was successfully prepared with excellent stability and sustained release for 24 h in vitro. The data of anti-inflammatory activity, asthma therapeutic effects and safety studies indicated that drug delivery mediated by liposomal nano-vehicles was a feasible and desirable strategy for medical strategy and showed great promise in the clinical therapy of asthma.

Integration of gut microbiome and serum metabolome revealed the effect of Qing-Wei-Zhi-Tong Micro-pills on gastric ulcer in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Qing-Wei-Zhi-Tong Micro-pills (QWZT) is herbal compound used in the treatment of GU, whose functions include clearing the stomach and fire, softening the liver and relieving pain. However, its mechanistic profile on host intestinal microbiota and metabolism has not been determined. AIM OF THE STUDY: The present study aimed to observe the healing effect of QWZT on acetic acid-induced gastric ulcer in a rat model and to preliminarily elucidate its possible therapeutic mechanism from the perspective of host intestinal microbiota and metabolism. MATERIALS AND METHODS: The Wistar male rats (7 weeks old; weight 180-200 g) were randomly divided into normal control group (NC), acetic acid-induced gastric ulcer group (GU), and QWZT treatment group (High dose: 1250 mg/kg/day, Middle dose: 625 mg/kg/day, Low dose: 312.5 mg/kg/day) of 6 rats each. An acetic acid-induced gastric ulcer rat model was constructed based on anatomical surgery. QWZT (High dose, Middle dose, and Low dose) was used to treat gastric ulcer rats for 7 days by gavage. At the end of treatment, the body weight, macroscopic condition of gastric tissue ulcers, pathological changes (HE staining), inflammatory factors, oxidative stress factors, and endocrine factors were assessed in each group of rats. Fresh feces and serum from each group of rats were collected for microbiome and metabolome analysis on the machine, respectively. Drug-disease common targets and functional pathways were captured based on network pharmacology. The complex network of Herbs-Targets-Pathways-Metabolites-Microbiota interactions was constructed. Ultimately, Fecal Microbiota Transplantation (FMT) evaluated the contribution of gut microbiota in disease. RESULTS: QWZT increased the abundance of beneficial bacteria (Bacteroides, Alloprevotella, Rikenellaceae_RC9_gut_group, Lactobacillus, Lachnospiraceae_NK4A136_group, Parabacteroides, etc.), reduced the abundance of harmful bacteria (Micromonospora, Geobacter, Nocardioides, and Arenimonas, etc.), reduced the levels of inflammatory mediators (12,13-EpOME, 9,10-Epoxyoctadecenoic acid, SM(d18:1/16:0) and Leukotriene A4, etc.), restored host metabolic disorders (Linoleic acid metabolism, Glycerophospholipid metabolism, and Arachidonic acid metabolism), and regulated the level of cytokines (IL-6, TNF-a, SOD, MDA, PEG-2 and NO), ultimately exerting an anti-ulcer effect. Apart from that, FMT improved acetic acid-induced gastric ulcers in rats. CONCLUSION: QWZT improved acetic acid-induced gastric ulcers in rats by remodeling intestinal microbiota and regulating host metabolism. This work may promote the process of developing and utilizing clinical applications of QWZT.

Enhanced Oxygen Evolution Rate and Anti-interfacial Delamination Property of the SrCo0.9Ta0.1O3-δ@La0.6Sr0.4Co0.2Fe0.8O3-δ Oxygen-Electrode for Solid Oxide Electrolysis Cells.

Interfacial delamination between the oxygen-electrode and electrolyte is a significant factor impacting the reliability of solid oxide electrolysis cells (SOECs) when operating at high voltages. The most effective method to mitigate this delamination is to decrease the interfacial oxygen partial pressure, which can be accomplished by amplifying the oxygen exsolution rate and the O2- transport rate of the oxygen-electrode. In this study, a SrCo0.9Ta0.1O3-δ (SCT) film with an outstanding oxygen surface exchange coefficient and an outstanding O2- conductivity was introduced onto the La0.6Sr0.4Co0.2Fe0.8O3-δ (LSCF) surface by infiltration. This composite oxygen-electrode exhibited a notably high electrochemical catalytic activity primarily due to the significantly improved O2- transport and oxygen surface exchange rate. Single cells with a 15-LSCF oxygen-electrode achieved a peak power density of 1.33 W cm-2 at 700 °C and a current density of 1.25 A cm-2 at 1.3 V (60% H2O-H2) at 750 °C. Additionally, an electrolysis cell with a 15 wt % SCT-infiltrated LSCF oxygen-electrode demonstrated stable operation even at high current densities for over 330 h with no noticeable delamination. The remarkable durability of the 15-LSCF oxygen-electrode can be attributed to the boosted oxygen exsolution reaction (OER) activity and the suppression of Sr segregation due to SCT infiltration. The impressive OER activity and resistance to interfacial delamination make the 15-LSCF a promising candidate for a composite oxygen-electrode in SOECs.

A Multiscale Point-Supervised Network for Counting Maize Tassels in the Wild.

Accurate counting of maize tassels is essential for monitoring crop growth and estimating crop yield. Recently, deep-learning-based object detection methods have been used for this purpose, where plant counts are estimated from the number of bounding boxes detected. However, these methods suffer from 2 issues: (a) The scales of maize tassels vary because of image capture from varying distances and crop growth stage; and (b) tassel areas tend to be affected by occlusions or complex backgrounds, making the detection inefficient. In this paper, we propose a multiscale lite attention enhancement network (MLAENet) that uses only point-level annotations (i.e., objects labeled with points) to count maize tassels in the wild. Specifically, the proposed method includes a new multicolumn lite feature extraction module that generates a scale-dependent density map by exploiting multiple dilated convolutions with different rates, capturing rich contextual information at different scales more effectively. In addition, a multifeature enhancement module that integrates an attention strategy is proposed to enable the model to distinguish between tassel areas and their complex backgrounds. Finally, a new up-sampling module, UP-Block, is designed to improve the quality of the estimated density map by automatically suppressing the gridding effect during the up-sampling process. Extensive experiments on 2 publicly available tassel-counting datasets, maize tassels counting and maize tassels counting from unmanned aerial vehicle, demonstrate that the proposed MLAENet achieves marked advantages in counting accuracy and inference speed compared to state-of-the-art methods. The model is publicly available at https://github.com/ShiratsuyuShigure/MLAENet-pytorch/tree/main.

Efferocytosis and Respiratory Disease.

Cells are the smallest units that make up living organisms, which constantly undergo the processes of proliferation, differentiation, senescence and death. Dead cells need to be removed in time to maintain the homeostasis of the organism and keep it healthy. This process is called efferocytosis. If the process fails, this may cause different types of diseases. More and more evidence suggests that a faulty efferocytosis process is closely related to the pathological processes of respiratory diseases. In this review, we will first introduce the process and the related mechanisms of efferocytosis of the macrophage. Secondly, we will propose some methods that can regulate the function of efferocytosis at different stages of the process. Next, we will discuss the role of efferocytosis in different lung diseases and the related treatment approaches. Finally, we will summarize the drugs that have been applied in clinical practice that can act upon efferocytosis, in order to provide new ideas for the treatment of lung diseases.

PLGA-Based Micro/Nanoparticles: An Overview of Their Applications in Respiratory Diseases.

Respiratory diseases, such as asthma and chronic obstructive pulmonary disease (COPD), are critical areas of medical research, as millions of people are affected worldwide. In fact, more than 9 million deaths worldwide were associated with respiratory diseases in 2016, equivalent to 15% of global deaths, and the prevalence is increasing every year as the population ages. Due to inadequate treatment options, the treatments for many respiratory diseases are limited to relieving symptoms rather than curing the disease. Therefore, new therapeutic strategies for respiratory diseases are urgently needed. Poly (lactic-co-glycolic acid) micro/nanoparticles (PLGA M/NPs) have good biocompatibility, biodegradability and unique physical and chemical properties, making them one of the most popular and effective drug delivery polymers. In this review, we summarized the synthesis and modification methods of PLGA M/NPs and their applications in the treatment of respiratory diseases (asthma, COPD, cystic fibrosis (CF), etc.) and also discussed the research progress and current research status of PLGA M/NPs in respiratory diseases. It was concluded that PLGA M/NPs are the promising drug delivery vehicles for the treatment of respiratory diseases due to their advantages of low toxicity, high bioavailability, high drug loading capacity, plasticity and modifiability. And at the end, we presented an outlook on future research directions, aiming to provide some new ideas for future research directions and hopefully to promote their widespread application in clinical treatment.

VgrG Spike Dictates PAAR Requirement for the Assembly of the Type VI Secretion System.

Widely employed by Gram-negative pathogens for competition and pathogenesis, the type six protein secretion system (T6SS) can inject toxic effectors into neighboring cells through the penetration of a spear-like structure comprising a long Hcp tube and a VgrG-PAAR spike complex. The cone-shaped PAAR is believed to sharpen the T6SS spear for penetration but it remains unclear why PAAR is required for T6SS functions in some bacteria but dispensable in others. Here, we report the conditional requirement of PAAR for T6SS functions in Aeromonas dhakensis, an emerging human pathogen that may cause severe bacteremia. By deleting the two PAAR paralogs, we show that PAAR is not required for T6SS secretion, bacterial killing, or specific effector delivery in A. dhakensis. By constructing combinatorial PAAR and vgrG deletions, we demonstrate that deletion of individual PAAR moderately reduced T6SS functions but double or triple deletions of PAAR in the vgrG deletion mutants severely impaired T6SS functions. Notably, the auxiliary-cluster-encoded PAAR2 and VgrG3 are less critical than the main-cluster-encoded PAAR1 and VgrG1&2 proteins to T6SS functions. In addition, PAAR1 but not PAAR2 contributes to antieukaryotic virulence in amoeba. Our data suggest that, for a multi-PAAR T6SS, the variable role of PAAR paralogs correlates with the VgrG-spike composition that collectively dictates T6SS assembly. IMPORTANCE Gram-negative bacteria often encode multiple paralogs of the cone-shaped PAAR that sits atop the VgrG-spike and is thought to sharpen the spear-like T6SS puncturing device. However, it is unclear why PAAR is required for the assembly of some but not all T6SSs and why there are multiple PAARs if they are not required. Our data delineate a VgrG-mediated conditional requirement for PAAR and suggest a core-auxiliary relationship among different PAAR-VgrG modules that may have been acquired sequentially by the T6SS during evolution.

Murine fertility and spermatogenesis are independent of the testis-specific Spdye4a gene.

BACKGROUND: While many testis-enriched genes have been identified as important regulators of the spermatogenic process, the specific roles played by several of these genes and their functional importance has yet to be fully clarified. METHODS: We employed a CRISPR/Cas9 approach to introduce a 5 bp in-frame deletion within the Spdye4a gene (Exon 2) of C57BL/6 mice (Spdye4a-/-). Fertility and sperm counts were evaluated. Testes tissues and cell suspensions were analyzed via histological and immunofluorescence staining. mRNA and protein levels of candidate genes were assessed through qPCR and Western blotting. In vitro fertilization was used to assess the ability of sperm cells to bind to egg cells. RESULTS: Spdye4a-/- mice did not exhibit any reduction in fertility, and exhibited comparable sperm counts, morphology and motility to those of wildtype littermates. Functionally, Spdye4a-/- sperm exhibited normal sperm-egg binding activity in vitro. Furthermore, the testes of Spdye4a-/- mice exhibited a full range of germ cells from spermatogonia to mature spermatozoa. No differences in the progression of meiotic prophase I were observed when comparing Spdye4a-/- and wildtype mice, indicating that the loss of Spdye4a had no adverse effect on spermatogenesis. DISCUSSION: Spdye4a is dispensable in the context of mice fertility and spermatogenesis. This study will prevent other laboratories from expending repeated efforts to generate similar knockout mice.

The combined action of biochar and nitrogen-fixing bacteria on microbial and enzymatic activities of soil N cycling.

This study aims to investigate the positive effects of the combined use of Enterobacter cloacae and biochar on improving nitrogen (N) utilization. The greenhouse pots experimental results showed the synergy of biochar and E. cloacae increased soil total N content and plant N uptake by 33.54% and 15.1%, respectively. Soil nitrogenase (NIT) activity increased by 253.02%. Ammonia monooxygenase (AMO) and nitrate reductase (NR) activity associated with nitrification and denitrification decreased by 10.94% and 29.09%, respectively. The relative abundance of N fixing microorganisms like Burkholderia and Bradyrhizobium significantly increased. Sphingomonas and Ottowia, two bacteria involved in the nitrification and denitrification processes, were found to be in lower numbers. The E. cloacae's ability to fix N2 and promote the growth of plants allow the retention of N in soil and make more N available for plant development. Biochar served as a reservoir of N for plants by adsorbing N from the soil and providing a shelter for E. cloacae. Thus, biochar and E. cloacae form a synergy for the management of agricultural N and the mitigation of negative impacts of pollution caused by excessive use of N fertilizer.

Recent Advances in Nanomaterials for Asthma Treatment.

Asthma is a chronic airway inflammatory disease with complex mechanisms, and these patients often encounter difficulties in their treatment course due to the heterogeneity of the disease. Currently, clinical treatments for asthma are mainly based on glucocorticoid-based combination drug therapy; however, glucocorticoid resistance and multiple side effects, as well as the occurrence of poor drug delivery, require the development of more promising treatments. Nanotechnology is an emerging technology that has been extensively researched in the medical field. Several studies have shown that drug delivery systems could significantly improve the targeting, reduce toxicity and improve the bioavailability of drugs. The use of multiple nanoparticle delivery strategies could improve the therapeutic efficacy of drugs compared to traditional delivery methods. Herein, the authors presented the mechanisms of asthma development and current therapeutic methods. Furthermore, the design and synthesis of different types of nanomaterials and micromaterials for asthma therapy are reviewed, including polymetric nanomaterials, solid lipid nanomaterials, cell membranes-based nanomaterials, and metal nanomaterials. Finally, the challenges and future perspectives of these nanomaterials are discussed to provide guidance for further research directions and hopefully promote the clinical application of nanotherapeutics in asthma treatment.

Heterologous Assembly of the Type VI Secretion System Empowers Laboratory Escherichia coli with Antimicrobial and Cell Penetration Capabilities.

The synthetic biology toolbox has amassed a vast number of diverse functional modules, but protein translocation modules for cell penetration and cytosol-to-cytosol delivery remain relatively scarce. The type VI secretion system (T6SS), commonly found in many Gram-negative pathogens, functions as a contractile device to translocate protein toxins to prokaryotic and eukaryotic cells. Here, we have assembled the T6SS of Aeromonas dhakensis, an opportunistic waterborne pathogen, in the common laboratory strain Escherichia coli BL21(DE3). We constructed a series of plasmids (pT6S) carrying the T6SS structural and effector genes under native or tetracycline-inducible promoters, the latter for controlled expression. Using fluorescence microscopy and biochemical analyses, we demonstrate a functional T6SS in E. coli capable of secreting proteins directly into the cytosol of neighboring bacteria and outcompeting a number of drug-resistant pathogens. The heterologous assembly of T6SS not only confers the lab workhorse E. coli with the cytosol-to-cytosol protein delivery capability but also demonstrates the potential for harnessing the T6SS of various pathogens for general protein delivery and antibacterial applications. IMPORTANCE The T6SS is a powerful and versatile protein delivery system. However, the complexity of its macromolecular structure and gene regulation makes it not a trivial task to reconstitute the T6SSs of pathogens in a nonpathogenic host. In this study, we have assembled an inducible T6SS in E. coli BL21(DE3) and demonstrated its functions in protein delivery and antimicrobial activities. The engineered T6SS empowers E. coli to deliver protein cargos into a wide range of prokaryotic and eukaryotic cells.

A Single Amino Acid Substitution in MIL1 Leads to Activation of Programmed Cell Death and Defense Responses in Rice.

Lesion mimic mutants are an ideal model system for elucidating the molecular mechanisms of programmed cell death and defense responses in rice. In this study, we identified a lesion mimic mutant termed miner infection like 1-1 (mil1-1). The mil1-1 exhibited lesions on the leaves during development, and the chloroplasts of mil1-1 leaves were disrupted. Reactive oxygen species were found to accumulate in mil1-1 leaves. Cell death and DNA fragmentation were observed in mil1-1 leaves, indicating that the cells in the spots of mil1-1 leaves experienced programmed cell death. Most agronomic traits decreased in mil1-1, suggesting that the growth retardation in mil1-1 caused reduced per-plant grain yield. However, the mutation of MIL1 activated the expression of pathogen response genes and enhanced resistance to bacterial blight. The MIL1 gene was cloned using the positional cloning approach. A missense mutation 751 bp downstream of ATG was found in mil1-1. The defects of mil1-1 were able to be rescued by delivering a wild-type MIL1 gene into mil1-1. MIL1 encoded hydroperoxide lyase 3 (OsHPL3), and the expression of OsHPL3 was induced via hormone and abiotic stresses. Our findings provide insights into the roles of MIL1 in regulating programmed cell death, development, yield, and defense responses in rice.

Downregulated Wnt2B Expression Suppresses Proliferation, Invasion, and Angiogenesis of Ovarian Cancer Cells Through Inhibiting the Wnt/ß-Catenin Signaling Pathway.

Ovarian cancer (OC) is known to be the most malignant gynecologic cancers. Wnt2B, a member of the Wnt family, plays a critical role in tumor development. However, the effect of Wnt2B on the occurrence and development of OC remains largely uncharacterized. In this study, immunohistochemistry assay indicated that Wnt2B was increased in our study cohort (OC). In addition, the expression of Wnt2B was positively correlated with TNM stages and metastasis of OC patients. Wnt2B markedly mediated the regulation of OC proliferation, invasion, and angiogenesis. Moreover, Wnt2B knockdown inactivated the Wnt/ß-catenin signaling pathway. More importantly, the Wnt/ß-catenin signaling pathway activator LiCl reversed the effect of Wnt2B knockdown on OC cell proliferation, angiogenesis, and invasion. Our data indicated that Wnt2B silencing could inhibit the proliferation, invasion, and angiogenesis of OC cells through downregulating the activity of Wnt/ß-catenin pathway.

MiR-125b-5p ameliorates hypoxia/reoxygenation-induced endothelial cell dysfunction and attenuates reduced uterine perfusion pressure-induced hypertension in pregnant rats via targeting BMF.

BACKGROUND AND PURPOSE: MicroRNA-125b-5p (miR-125b-5p) is downregulated in patients with gestational hypertension signs. However, the role of miR-125b-5p in pregnancy-induced hypertension (PIH) remains unknown. METHODS: The human placental microvascular endothelial cells (HPMECs) have undergone hypoxia and reoxygenation (H/R) treatment to establish PIH cellular model. Rats were performed with reduced uterine perfusion pressure (RUPP) operation to establish PIH animal model. RESULTS: MiR-125b-5p promoted viability while inhibited the apoptosis of H/R-treated HPMECs by downregulating BMF. MiR-125b-5p alleviated hypertensive symptoms and improved pregnancy outcomes in RUPP rats. CONCLUSION: MiR-125b-5p ameliorates H/R-induced HPMEC dysfunction and attenuates RUPP-induced hypertension in pregnant rats by downregulating BMF.

Dysregulated pseudogene BNIP3P1 inhibited cell proliferation and promoted cell apoptosis in preeclampsia by acting as a competing endogenous RNA for BNIP3.

Long noncoding RNAs (lncRNAs) have been reported as critical modulators in many diseases including preeclampsia. Since the association between lncRNA BNIP3P1 and its cognate gene BNIP3 in preeclampsia has been revealed previously, this study aimed to further explore the function and mechanism of BNIP3P1 in preeclampsia. EdU and TUNEL assays revealed that BNIP3P1 or BNIP3 overexpression inhibited trophoblast cell proliferation and enhanced cell apoptosis in preeclampsia. As suggested by western blot analysis, the protein levels of apoptotic markers in the cells were affected by BNIP3P1 or BNIP3 overexpression. The binding between miR-128-3p and BNIP3P1 (or BNIP3) was explored by luciferase reporter assays. Mechanistically, BNIP3P1 bound to miR-128-3p to upregulate BNIP3 expression by acting as a competing endogenous RNA (ceRNA). Importantly, BNIP3P1 was found to inactivate the mTOR signaling pathway. In conclusion, BNIP3P1 inhibited trophoblast cell proliferation and enhanced cell apoptosis in preeclampsia by targeting the miR-128-3p/BNIP3/mTOR signaling pathway.

MicroRNA-195-5p facilitates endothelial dysfunction by inhibiting vascular endothelial growth factor A in gestational diabetes mellitus.

Gestational diabetes mellitus (GDM) is a common disorder during pregnancy associated with endothelial dysfunction in the placental vasculature. MicroRNAs (miRNAs), which are short noncoding RNAs that modulate post-transcriptional gene expression, affect GDM progression. MiR-195-5p was reported to be a putative biomarker for GDM diagnosis, whose expression was markedly elevated in serum of GDM patients. Therefore, our study intended to explore whether miR-195-5p regulates endothelial cell dysfunction in GDM. Human placental microvascular endothelial cells (hPMECs) were treated with high concentration of glucose to establish an in vitro GDM model. The apoptosis, proliferation and angiogenesis of hPMECs were detected by flow cytometry analysis, CCK-8 assay and tube formation assay. The binding between vascular endothelial growth factor A (VEGFA) and miR-195-5p was verified by luciferase reporter assay. GDM mouse model was established by intraperitoneal injection of streptozocin. Cell apoptosis and the pathological changes in GDM mouse placenta tissues were evaluated by TUNEL staining and HE staining. Gene expression was detected by RT-qPCR. Protein levels were evaluated by western blotting. In this study, miR-195-5p knockdown promoted the proliferation and angiogenesis as well as inhibited the apoptosis of HG-treated hPMECs. MiR-195-5p targeted VEGFA, whose expression was downregulated in HG-treated hPMECs. VEGFA silencing antagonized the influence of miR-195-5p knockdown on the phenotypes of HG-treated hPMECs. Additionally, miR-195-5p inhibition decelerated cell apoptosis and improved pathological changes in GDM mouse placenta tissues. MiR-195-5p level was negatively correlated to VEGFA level in GDM mouse placenta tissues. Overall, miR-195-5p facilitates the endothelial cell dysfunction by inhibiting VEGFA in GDM.

Development and Validation of Noninvasive MRI-Based Signature for Preoperative Prediction of Early Recurrence in Perihilar Cholangiocarcinoma.

BACKGROUND: Cholangiocarcinoma is a type of hepatobiliary tumor. For perihilar cholangiocarcinoma (pCCA), patients who experience early recurrence (ER) have a poor prognosis. Preoperative accurate prediction of postoperative ER can avoid unnecessary operation; however, prediction is challenging. PURPOSE: To develop a novel signature based on clinical and/or MRI radiomics features of pCCA to preoperatively predict ER. STUDY TYPE: Retrospective. POPULATION: One hundred eighty-four patients (median age, 61.0 years; interquartile range: 53.0-66.8 years) including 115 men and 69 women. FIELD STRENGTH/SEQUENCE: A 1.5 T; volumetric interpolated breath-hold examination (VIBE) sequence. ASSESSMENT: The models were developed from the training set (128 patients) and validated in a separate testing set (56 patients). The contrast-enhanced arterial and portal vein phase MR images of hepatobiliary system were used for extracting radiomics features. The correlation analysis, least absolute shrinkage and selection operator (LASSO) logistic regression (LR), backward stepwise LR were mainly used for radiomics feature selection and modeling (Modelradiomic ). The univariate and multivariate backward stepwise LR were used for preoperative clinical predictors selection and modeling (Modelclinic ). The radiomics and preoperative clinical predictors were combined by multivariate LR method to construct clinic-radiomics nomogram (Modelcombine ). STATISTICAL TESTS: Chi-squared (χ2 ) test or Fisher's exact test, Mann-Whitney U-test or t-test, Delong test. Two tailed P < 0.05 was considered statistically significant. RESULTS: Based on the comparison of area under the curves (AUC) using Delong test, Modelclinic and Modelcombine had significantly better performance than Modelradiomic and tumor-node-metastasis (TNM) system in training set. In the testing set, both Modelclinic and Modelcombine had significantly better performance than TNM system, whereas only Modelcombine was significantly superior to Modelradiomic . However, the AUC values were not significantly different between Modelclinic and Modelcombine (P = 0.156 for training set and P = 0.439 for testing set). DATA CONCLUSION: A noninvasive model combining the MRI-based radiomics signature and clinical variables is potential to preoperatively predict ER for pCCA. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY STAGE: 4.