Sub-genomic RNAi-assisted strain evolution of filamentous fungi for enhanced protein production.

Genetic engineering at the genomic scale provides a rapid means to evolve microbes for desirable traits. However, in many filamentous fungi, such trials are daunted by low transformation efficiency. Differentially expressed genes under certain conditions may contain important regulatory factors. Accordingly, although manipulating these subsets of genes only can largely reduce the time and labor, engineering at such a sub-genomic level may also be able to improve the microbial performance. Herein, first using the industrially important cellulase-producing filamentous fungus Trichoderma reesei as a model organism, we constructed suppression subtractive hybridization (SSH) libraries enriched with differentially expressed genes under cellulase induction (MM-Avicel) and cellulase repression conditions (MM-Glucose). The libraries, in combination with RNA interference, enabled sub-genomic engineering of T. reesei for enhanced cellulase production. The ability of T. reesei to produce endoglucanase was improved by 2.8~3.3-fold. In addition, novel regulatory genes (tre49304, tre120391, and tre123541) were identified to affect cellulase expression in T. reesei. Iterative manipulation using the same strategy further increased the yield of endoglucanase activity to 75.6 U/mL, which was seven times as high as that of the wild type (10.8 U/mL). Moreover, using Humicola insolens as an example, such a sub-genomic RNAi-assisted strain evolution proved to be also useful in other industrially important filamentous fungi. H. insolens is a filamentous fungus commonly used to produce catalase, albeit with similarly low transformation efficiency and scarce knowledge underlying the regulation of catalase expression. By combining SSH and RNAi, a strain of H. insolens producing 28,500 ± 288 U/mL of catalase was obtained, which was 1.9 times as high as that of the parent strain.IMPORTANCEGenetic engineering at the genomic scale provides an unparalleled advantage in microbial strain improvement, which has previously been limited only to the organisms with high transformation efficiency such as Saccharomyces cerevisiae and Escherichia coli. Herein, using the filamentous fungus Trichoderma reesei as a model organism, we demonstrated that the advantage of suppression subtractive hybridization (SSH) to enrich differentially expressed genes and the convenience of RNA interference to manipulate a multitude of genes could be combined to overcome the inadequate transformation efficiency. With this sub-genomic evolution strategy, T. reesei could be iteratively engineered for higher cellulase production. Intriguingly, Humicola insolens, a fungus with even little knowledge in gene expression regulation, was also improved for catalase production. The same strategy may also be expanded to engineering other microorganisms for enhanced production of proteins, organic acids, and secondary metabolites.

Two novel CHN1 variants identified in Duane retraction syndrome pedigrees disrupt development of ocular motor nerves in zebrafish.

Duane retraction syndrome (DRS) is a rare congenital eye movement disorder causing by the dysplasia of abducens nerve, and has highly variable phenotype. MRI can reveal the endophenotype of DRS. Most DRS cases are sporadical and isolated, while some are familial or accompanied by other ocular disorders and systemic congenital abnormalities. CHN1 was the most common causative gene for familial DRS. Until now, 13 missense variants of CHN1 have been reported. In this study, we enrolled two unrelated pedigrees with DRS. Detailed clinical examinations, MRI, and the whole exome sequencing (WES) were performed to reveal their clinical and genetic characteristics. Patients from pedigree-1 presented with isolated DRS, and a novel heterozygous variant c.650 A > G, p. His217Arg was identified in CHN1 gene. Patients from pedigree-2 presented with classic DRS and abnormalities in auricle morphology, and the pedigree segregated another novel heterozygous CHN1 variant c.637 T > C, p. Phe213Leu. A variety of bioinformatics software predicted that the two variants had deleterious or disease-causing effects. After injecting of two mutant CHN1 mRNAs into zebrafish embryos, the dysplasia of ocular motor nerves (OMN) was observed. Our present findings expanded the phenotypic and genotypic spectrum of CHN1 related DRS, as well as provided new insights into the role of CHN1 in OMN development. Genetic testing is strongly recommended for patients with a DRS family history or accompanying systemic congenital abnormalities.

Preparation and indoor virulence determination of a temperature-sensitive insecticide against Zeugodacus cucurbitae (Coquillett).

Zeugodacus cucurbitae (Coquillett) is an important fruit and vegetable pest, especially in high-temperature seasons. In our previous research, we developed a temperature-sensitive sustained-release attractant for Z. cucurbitae, that not only can control the release rate of cuelure according to the temperature change, but also shows an excellent trapping effect on Z. cucurbitae. To further enhance the killing effect of the temperature-sensitive attractant on Z. cucurbitae, this study proposed using it in combination with an insecticide to prepare a temperature-sensitive insecticide for Z. cucurbitae. Based on the controlled release technology of pesticides, a temperature-sensitive Z. cucurbitae insecticide was developed by using PNIPAM gel as a temperature-sensitive switch to carry both cuelure and insecticide at the same time. In addition, the lethal effect of different pesticides on Z. cucurbitae were tested by indoor toxicity test, and the best pesticide combination was screened out. The temperature-sensitive insecticide prepared in this study not only had excellent thermal response and controlled release ability, but also enhanced its toxicological effects on Z. cucurbitae because it contained insecticides. Among them, combining thiamethoxam and clothianidin with the temperature-sensitive attractants was the most effective, and their lethality reached more than 97% against Z. cucurbitae. This study is not only of great practical significance for the monitoring and controlling Z. cucurbitae, but also provides theoretical basis and reference value for the combination of temperature-sensitive attractant and insecticide.

Longitudinal analysis of quality of life in primary lung cancer patients with chlamydia pneumoniae infection: a time-to-deterioration model.

BACKGROUND: Chlamydia pneumoniae (Cpn) IgG and IgA has been strongly linked to lung cancer, but its impact on patients' quality of life remains unclear. Our objective was to investigate the relationship between pre-treatment Cpn IgG and IgA and time to deterioration (TTD) of the HRQoL in patients with primary lung cancer. METHODS: A prospective hospital-based study was conducted from June 2017 to December 2018, enrolling 82 patients with primary lung cancer admitted to the First Affiliated Hospital of Fujian Medical University for questionnaire surveys. Cpn IgG and IgA was detected by microimmunofluorescence method. HRQoL was assessed at baseline and during follow-up using the EORTC Quality of Life Questionnaire version 3.0 (EORTC QLQ-C30) and EORTC Quality of Life Questionnaire-Lung Cancer (EORTC QLQ-LC13). HRQoL scores were calculated using the QoLR package, and TTD events were determined (minimum clinically significant difference = 5 points). Cox regression analysis was used to evaluate the effect of Cpn IgG and IgA on HRQoL. RESULTS: We investigated the relationship between Cpn IgG and IgA and quality of life in patients with primary lung cancer. The study was found that 75.61% of cases were Cpn IgG + and 45.12% were Cpn IgA + . Cpn IgA + IgG + was 41.46%. For EORTC QLQ-C30, Physical function (PF) and Pain (PA) TTD events on the functional scale and Symptom scale were the most common during follow-up. After adjusting for gender and smoking status, Pre-treatment Cpn IgA + was found to signifcantly delay TTD of Physical functioning(HR = 0.539, 95% CI: 0.291-0.996, P = 0.048). In addition, Cpn IgG + before treatment significantly delayed TTD in Emotional functioning (HR = 0.310, 95% CI: 0.115-0.836, P = 0.021). For EORTC QLQ-LC13, deterioration of dyspnea (LC-DY) was the most common event. However, Cpn IgG and IgA before treatment had no effect on the TTD of EORTC QLQ-LC13 items. CONCLUSIONS: According to EORTC QLQ-C30 and EORTC QLQ-LC13, Cpn IgA delayed TTD in Physical functioning and Cpn IgG delayed TTD in Emotional functioning.

Childhood adversity, perceived social support, and depressive symptoms among pre-abortion Chinese women.

BACKGROUND: Unintended (unwanted) pregnancy is a sexual and reproductive health issue with psychosocial consequences for the individual, their family, and society. However, the relationship between social support and related mental health issues, like depression and the effects of childhood adversity, is poorly studied. This study aims to explore the connections between childhood adversity, perceived social support, and depressive symptoms in pre-abortion women (women who have decided to have an abortion) in a clinical setting, based on the common risk factor approach and social support theory. METHODS: A total of 299 pre-abortion Chinese women 18-45 years were recruited in a hospital in Shantou, China. Hierarchical linear regression analyses were employed to examine the relative effects of childhood adversity and sources of social support on depressive symptoms, controlling for sociodemographic influences. RESULTS: The results show that 37.2 percent of participants reported at least one adverse experience in childhood. More than half of the respondents were at risk for depression. Results of regression analysis showed that childhood adversities were negatively associated with depressive symptoms before sources of social support were entered into the model. However, when the sources of perceived social support were added, the effect of childhood adversity was not significant. Perceived social support explained the additional 15 percent variance in depressive symptoms. Additionally, being married (ß = -.12, p < .05) and number of siblings (ß = .13, p < .05) were significantly related to depressive symptoms. DISCUSSION: Pre-abortion women are at risk of mental health problems. Peer and familial social supports can alleviate the influence of childhood adversity on depression among pre-abortion Chinese women. Strengthening the role of various sources of social support can help to improve the mental health conditions of pre-abortion women.

Preparation and Photocatalytic Properties of Al2O3-SiO2-TiO2 Porous Composite Semiconductor Ceramics.

Titanium dioxide (TiO2) is widely employed in the catalytic degradation of wastewater, owing to its robust stability, superior photocatalytic efficiency, and cost-effectiveness. Nonetheless, isolating the fine particulate photocatalysts from the solution post-reaction poses a significant challenge in practical photocatalytic processes. Furthermore, these particles have a tendency to agglomerate into larger clusters, which diminishes their stability. To address this issue, the present study has developed Al2O3-SiO2-TiO2 composite semiconductor porous ceramics and has systematically explored the influence of Al2O3 and SiO2 on the structure and properties of TiO2 porous ceramics. The findings reveal that the incorporation of Al2O3 augments the open porosity of the ceramics and inhibits the aggregation of TiO2, thereby increasing the catalytic site and improving the light absorption capacity. On the other hand, the addition of SiO2 enhances the bending strength of the ceramics and inhibits the conversion of anatase to rutile, thereby further enhancing its photocatalytic activity. Consequently, at an optimal composition of 55 wt.% Al2O3, 40 wt.% TiO2, and 5 wt.% SiO2, the resulting porous ceramics exhibit a methylene blue removal rate of 91.50%, and even after undergoing five cycles of testing, their catalytic efficiency remains approximately 83.82%. These outcomes underscore the exceptional photocatalytic degradation efficiency, recyclability, and reusability of the Al2O3-SiO2-TiO2 porous ceramics, suggesting their substantial potential for application in the treatment of dye wastewater, especially for the removal of methylene blue.

Extracellular vesicle-packaged miR-181c-5p from epithelial ovarian cancer cells promotes M2 polarization of tumor-associated macrophages via the KAT2B/HOXA10 axis.

OBJECTIVES: The molecular mechanistic actions of tumor-derived extracellular vesicles (EVs) in modulating macrophage polarization in the tumor microenvironment of epithelial ovarian cancer (EOC) is largely unknown. The study was performed to clarify the effect and downstream mechanism of microRNA-181c-5p (miR-181c-5p)-containing EVs from EOC cells in the M2 polarization of tumor-associated macrophages (TAMs). METHODS: EVs were isolated from normoxic and hypoxic human EOC cells SKOV3. Human mononuclear cell THP-1 was induced by phorbol-12-myristate-13-acetate to differentiate into TAMs. The targeting relationship between miR-181c-5p and KAT2B was verified by dual luciferase reporter gene assay. The interaction between KAT2B and HOXA10 was detected by immunofluorescence, Co-IP and ChIP assays. EdU staining, the scratch test and Transwell assay were used to assess the resultant cell proliferation, migration and invasion. The mouse xenograft model and the pulmonary metastasis model were developed through intraperitoneal injection of SKOV3 cells and tail vein injection of THP-1 cells, respectively. RESULTS: Hypoxic SKOV3 cell-derived EVs could be internalized by TAMs. SKOV3 cell-derived EVs induced by hypoxia (H-EVs) promoted M2 polarization of TAMs and facilitated the proliferation, migration and invasion of SKOV3 cells. miR-181c-5p was highly expressed in H-EVs and promoted the M2 polarization of TAMs. Further, miR-181c-5p targeted KAT2B, upregulated HOXA10 and activated the JAK1/STAT3 pathway, thereby promoting the M2 polarization of TAMs. In both mouse models, H-EV-derived miR-181c-5p promoted growth and metastasis of EOC cells. CONCLUSION: The miR-181c-5p-containing EVs from hypoxic EOC cells may upregulate HOXA10 by targeting KAT2B and activate the JAK1/STAT3 pathway to promote the M2 polarization of TAMs, ultimately promoting growth and metastasis of EOC cells in vitro and in vivo.

Construction of an immune-related lncRNA signature pair for predicting oncologic outcomes and the sensitivity of immunosuppressor in treatment of lung adenocarcinoma.

BACKGROUND: Although immunotherapy has shown clinical activity in lung adenocarcinoma (LUAD), LUAD prognosis has been a perplexing problem. We aimed to construct an immune-related lncRNA pairs (IRLPs) score for LUAD and identify what immunosuppressor are appropriate for which group of people with LUAD. METHODS: Based on The Cancer Genome Atlas (TCGA)-LUAD cohort, IRLPs were identified to construct an IRLPs scoring system by Cox regression and validated in the Gene Expression Omnibus (GEO) dataset using log-rank test and the receiver operating characteristic curve (ROC). Next, we used spearman's correlation analysis, t-test, signaling pathways analysis and gene mutation analysis to explore immune and molecular characteristics in different IRLP subgroups. The "pRRophetic" package was used to predict the sensitivity of immunosuppressant. RESULTS: The IRLPs score was constructed based on eight IRLPs calculated as 2.12 × (MIR31HG|RRN3P2) + 0.43 × (NKX2-1-AS1|AC083949.1) + 1.79 × (TMPO-AS1|LPP-AS2) + 1.60 × (TMPO-AS1|MGC32805) + 1.79 × (TMPO-AS1|PINK1-AS) + 0.65 × (SH3BP5-AS1|LINC01137) + 0.51 × (LINC01004|SH3PXD2A-AS1) + 0.62 × (LINC00339|AGAP2-AS1). Patients with a lower IRLPs risk score had a better overall survival (OS) (Log-rank test P TCGA train dataset < 0.001, P TCGA test dataset = 0.017, P GEO dataset = 0.027) and similar results were observed in the AUCs of TCGA dataset and GEO dataset (AUC TCGA train dataset = 0.777, AUC TCGA test dataset = 0.685, AUC TCGA total dataset = 0.733, AUC GEO dataset = 0.680). Immune score (Cor = -0.18893, P < 0.001), stoma score (Cor = -0.24804, P < 0.001), and microenvironment score (Cor = -0.22338, P < 0.001) were significantly decreased in the patients with the higher IRLP risk score. The gene set enrichment analysis found that high-risk group enriched in molecular changes in DNA and chromosomes signaling pathways, and in this group the tumor mutation burden (TMB) was higher than in the low-risk group (P = 0.0015). Immunosuppressor methotrexate sensitivity was higher in the high-risk group (P = 0.0052), whereas parthenolide (P < 0.001) and rapamycin (P = 0.013) sensitivity were lower in the high-risk group. CONCLUSIONS: Our study established an IRLPs scoring system as a biomarker to help in the prognosis, the identification of molecular and immune characteristics, and the patient-tailored selection of the most suitable immunosuppressor for LUAD therapy.

Folate deficiency disturbs PEG10 methylation modifications in human spina bifida.

BACKGROUND: Paternally expressed gene 10 (PEG10) is believed to be a key imprinted gene involved in placenta formation. However, its role in human folate-related spina bifida (SB) remains unclear. METHODS: The methylation status of the germline differentially methylated region (gDMR) in the PEG10/sarcoglycan epsilon (SGCE) imprinted cluster was compared between SB patients and control samples. Moreover, the influence of ectopic PEG10 expression on apoptosis was assessed to explore the underlying mechanisms related to folate deficiency-induced aberrant gDMR methylation in SB. RESULTS: The case group exhibited a significant increase in the methylation level of the gDMR and a marked reduction in the mRNA and protein expression of PEG10 compared with the control group. A prominent negative correlation was found between the folate level in brain tissue and gDMR methylation status (r = -0.62, P = 0.001). A cell model treated with a demethylating agent showed a significant elevation of PEG10 transcription level, as well as other imprinted genes in this cluster. In addition, the inhibition of PEG10 was found to be accompanied by aberrant activation of apoptosis in SB. CONCLUSIONS: Our findings suggest that disturbed gDMR methylation of the PEG10/SGCE cluster due to folate deficiency is involved in SB through aberrant activation of apoptosis. IMPACT: Disturbed genomic imprinting has been verified to be involved in neural tube defects (NTDs). However, little is known about the effect of ectopic expression of imprinted gene PEG10 on human NTDs. Aberrant methylation status of the germline differentially methylated region (gDMR) of PEG10/SGCE cluster due to folate deficiency has been found to result in the inhibition of PEG10 and has a marked association with an increased occurrence of spina bifida. Inhibited expression of PEG10 partly is found to be related to the abnormal activation of apoptosis in spina bifida.

BMP signaling is required for nkx2.3-positive pharyngeal pouch progenitor specification in zebrafish.

Pharyngeal pouches, a series of outpocketings that bud from the foregut endoderm, are essential to the formation of craniofacial skeleton as well as several important structures like parathyroid and thymus. However, whether pharyngeal pouch progenitors exist in the developing gut tube remains unknown. Here, taking advantage of cell lineage tracing and transgenic ablation technologies, we identified a population of nkx2.3+ pouch progenitors in zebrafish embryos and demonstrated an essential requirement of ectodermal BMP2b for their specification. At early somite stages, nkx2.3+ cells located at lateral region of pharyngeal endoderm give rise to the pouch epithelium except a subpopulation expressing pdgfαa rather than nkx2.3. A small-scale screen of chemical inhibitors reveals that BMP signaling is necessary to specify these progenitors. Loss-of-function analyses show that BMP2b, expressed in the pharyngeal ectoderm, actives Smad effectors in endodermal cells to induce nkx2.3+ progenitors. Collectively, our study provides in vivo evidence for the existence of pouch progenitors and highlights the importance of BMP2b signaling in progenitor specification.

Impact of Subjective and Objective Sleep Quality on Peptic Ulcer Rebleeding in Older Adults.

OBJECTIVE: This study aimed to examine the association of subjective and objective sleep quality with subsequent peptic ulcer rebleeding among older patients. METHODS: Of 1196 older patients with peptic ulcer bleeding (PUB) recruited from 12 grade A hospitals in the People's Republic of China, 1106 achieved full recovery from PUB, and they were followed up for up to 30 days. Using multiple measures at 1-week intervals, patients who presented PUB symptoms were invited to have an esophagogastroduodenoscopy examination. Subjective sleep quality was measured using the Pittsburgh Sleep Quality Index. Objective sleep quality domains were measured using an accelerometer, including sleep onset latency, sleep efficiency, total sleep time, and the number of awakenings. RESULTS: This study documented a 30-day cumulative ulcer rebleeding rate of 15.3%. Multivariate analyses showed that longer sleep onset latency (hazard ratio [HR] = 2.136 [1.336-2.558]) and more nighttime awakenings (HR = 1.698 [1.169-2.666]) increased the risk of ulcer rebleeding. However, a longer total sleep time (HR = 0.768 [0.698-0.887]) and better sleep efficiency (HR = 0.795 [0.682-0.975]) protected against ulcer rebleeding. Older patients who perceived poorer sleep quality were also more likely to experience ulcer rebleeding (HR = 2.295 [1.352-3.925]). CONCLUSIONS: The present results highlight the importance of proper treatment and prevention of sleep problems in older adults after successful PUB treatment. Our results, if replicable in future studies with more rigorous design and representative samples, might shed light on the etiology of ulcer rebleeding and suggest new pathways for preventing this disease.

Impact of Social Isolation on Subsequent Peptic Ulcer Recurrence in Older Adults With Mild Cognitive Impairment: The Role of Change in Severity of Depression.

OBJECTIVE: This study aimed to examine the association between social isolation, change in severity of depression, and subsequent recurrence of peptic ulcer disease (PUD) in older adults with mild cognitive impairment. METHODS: Older adults (≥55 years) with mild cognitive impairment and Helicobacter pylori-infected PUD (N = 2208) were recruited between 2010 and 2014 from 12 hospitals in the People's Republic of China. H. pylori was eradicated and PUD was cleared in 2015 participants by the end of 2014; 1900 of these were followed up for up to 36 months. The Kaplan-Meier method was used to assess how PUD recurrence varied with social engagement levels and changes in depression severity. Multivariate Cox proportional hazard models were used to examine associations between social isolation, changes in depression severity, and PUD recurrence. RESULTS: PUD recurrence was more prevalent in socially isolated (10.8%) than in socially engaged participants (5.5%). However, the rates of PUD were lower in socially isolated individuals without (absence of) depression (7.2%) and those with decreased depression (8.2%), whereas socially isolated individuals with unchanged and increased depression had substantially higher rates of PUD (16.3% and 17.8%, respectively; the social isolation by depression group for PUD recurrence was significant (p < .001). Specifically, although social isolation was associated with PUD recurrence during the 36-month follow-up period (hazard ratio [HR] = 2.665 [1.602-4.518]), it did not increase PUD recurrence risk in participants without depression or with reduced depression. However, in participants with unchanged or increased depression, PUD recurrence was more likely to occur in socially isolated (HR = 1.587 [1.125-2.588]; HR = 1.886 [1.012, 3.522] respectively) than in socially engaged participants. CONCLUSIONS: Social isolation is associated with a greater risk of PUD recurrence; however, the absence of or decreased severity of depression may alter this relationship.

Native T1 mapping compared to ultrasound elastography for staging and monitoring liver fibrosis: an animal study of repeatability, reproducibility, and accuracy.

OBJECTIVES: To investigate the repeatability, reproducibility, and staging and monitoring of the performance of native T1 mapping for noninvasively assessing liver fibrosis in comparison with acoustic radiation force impulse (ARFI) elastography. METHODS: The repeatability and reproducibility were explored in 8 male Sprague-Dawley rats with intraclass correlation coefficient (ICC). Different degrees of fibrosis were induced in 52 rats by carbon-tetrachloride (CCl4) insult. Another 16 rats were used to build fibrosis progression and regression models. The native T1 values and shear wave velocity (SWV) were quantified by using native T1 mapping and ARFI elastography, respectively. The METAVIR system (F0-F4) was used for the staging of fibrosis. The area under the receiver operating characteristic curve (AUC) was determined to assess the performance of quantitative parameters for staging and monitoring fibrosis. RESULTS: Native T1 values shared similar good repeatability (ICC = 0.93) and reproducibility (ICC = 0.87) with SWV (ICC = 0.84-0.93). The AUC of native T1 values were 0.84, 0.84, and 0.75 for diagnosing significant fibrosis (≥ F2) and liver cirrhosis (F4) and detecting fibrosis progression, and those of SWV were 0.81, 0.86, and 0.7, respectively. No significant difference in performance was found between the two quantitative parameters (p ≥ 0.496). For detecting fibrosis regression, native T1 values had a better accuracy (AUC = 0.99) than SWV (AUC = 0.56; p = 0.002). CONCLUSION: Native T1 mapping may be a reliable and accurate method for noninvasively assessing liver fibrosis. Compared with ARFI elastography, it provides similar good repeatability and reproducibility, a similar high accuracy for staging fibrosis, and a better accuracy for detecting fibrosis regression. KEY POINTS: • Native T1 mapping is a valuable tool for noninvasively assessing liver fibrosis and can be measured on virtually all clinical MRI machines without additional hardware or gadolinium chelate injection. • Compared with acoustic radiation force impulse elastography, native T1 mapping yields similar good repeatability and reproducibility and a similar high accuracy for staging fibrosis. • Native T1 mapping provides a significantly better performance for detecting fibrosis regression than acoustic radiation force impulse elastography.

A GEF activity-independent function for nuclear Net1 in Nodal signal transduction and mesendoderm formation.

Net1 is a well-characterized oncoprotein with RhoA-specific GEF activity. Oncogenic Net1, which lacks the first 145 amino acids, is present in the cytosol and contributes to the efficient activation of RhoA and the formation of actin stress fibers in a number of tumor cell types. Meanwhile, wild-type Net1 is predominantly localized in the nucleus at steady state due to its N-terminal nuclear localization sequences, where the function of nuclear Net1 has not been fully determined. Here, we find that zebrafish net1 is expressed specifically in mesendoderm precursors during gastrulation. Endogenous Net1 is located in the nucleus during early embryonic development. Gain- and loss-of-function experiments in zebrafish embryos and mammalian cells demonstrate that, regardless of its GEF activity, nuclear Net1 is critical for zebrafish mesendoderm formation and Nodal signal transduction. Detailed analyses of protein interactions reveal that Net1 associates with Smad2 in the nucleus in a GEF-independent manner, and then promotes Smad2 activation by enhancing recruitment of p300 (also known as EP300) to the transcriptional complex. These findings describe a novel genetic mechanism by which nuclear Net1 facilitates Smad2 transcriptional activity to guide mesendoderm development.

Rationale and design of the Henan ST elevation myocardial infarction (STEMI) registry: a regional STEMI project in predominantly rural central China.

BACKGROUND: Cardiovascular disease including ST elevation myocardial infarction (STEMI) is increasing and the leading cause of death in China. There has been limited data available to characterize STEMI management and outcomes in rural areas of China. The Henan STEMI Registry is a regional STEMI project with the objectives to timely obtain real-world knowledge about STEMI patients in secondary and tertiary hospitals and to provide a platform for care quality improvement efforts in predominantly rural central China. METHODS: The Henan STEMI Registry is a multicentre, prospective and observational study for STEMI patients. The registry includes 66 participating hospitals (50 secondary hospitals; 16 tertiary hospitals) that cover 15 prefectures and one city direct-controlled by the province in Henan province. Patients were consecutively enrolled with a primary diagnosis of STEMI within 30 days of symptom onset. Clinical treatments, outcomes and cost are collected by local investigators and captured electronically, with a standardized set of variables and standard definitions, and rigorous data quality control. Post-discharge patient follow-up to 1 year is planned. As of August 2018, the Henan STEMI Registry has enrolled 5479 patients of STEMI. DISCUSSION: The Henan STEMI Registry represents the largest Chinese regional platform for clinical research and care quality improvement for STEMI. The board inclusion of secondary hospitals in Henan province will allow for the exploration of STEMI in predominantly rural central China. TRIAL REGISTRATION: [NCT02641262] [29 December, 2015].

18F-labeled magnetic nanoparticles for monitoring anti-angiogenic therapeutic effects in breast cancer xenografts.

PURPOSE: To develop a novel fluorine-18 (18F)-labeled arginine-glycine-aspartic acid (RGD)-coupled ultra-small iron oxide nanoparticle (USPIO) (hereafter, referred to as 18F-RGD@USPIO) and conduct an in-depth investigation to monitor the anti-angiogenic therapeutic effects by using a novel dual-modality PET/MRI probe. METHODS: The RGD peptide and 18F were coupled onto USPIO by click chemistry. In vitro experiments including determination of stability, cytotoxicity, cell binding of the obtained 18F-RGD@USPIO were carried out, and the targeting kinetics and bio-distribution were tested on an MDA-MB-231 tumor model. A total of 20 (n = 10 per group) MDA-MB-231 xenograft-bearing mice were treated with bevacizumab or placebo (intraperitoneal injections of bevacizumab or a volume-equivalent placebo solution at the dose of 5 mg/kg for consecutive 7 days, respectively), and underwent PET/CT and MRI examinations with 18F-RGD@USPIO before and after treatment. Imaging findings were validated by histological analysis with regard to ß3-integrin expression (CD61 expression), microvascular density (CD31 expression), and proliferation (Ki-67 expression). RESULTS: Excellent stability, low toxicity, and good specificity to endothelial of 18F-RGD@USPIO were confirmed. The best time point for MRI scan was 6 h post-injection. No intergroup differences were observed in tumor volume development between baseline and day 7. However, 18F-RGD@USPIO binding was significantly reduced after bevacizumab treatment compared with placebo, both on MRI (P < 0.001) and PET/CT (P = 0.002). Significantly lower microvascular density, tumor cell proliferation, and integrin ß3 expression were noted in the bevacizumab therapy group than the placebo group, which were consistent with the imaging results. CONCLUSION: PET/MRI with the dual-modality nanoprobe, 18F-RGD@USPIO, can be implemented as a noninvasive approach to monitor the therapeutic effects of anti-angiogenesis in breast cancer model in vivo.

Activation of Smurf E3 ligase promoted by smoothened regulates hedgehog signaling through targeting patched turnover.

Hedgehog signaling plays conserved roles in controlling embryonic development; its dysregulation has been implicated in many human diseases including cancers. Hedgehog signaling has an unusual reception system consisting of two transmembrane proteins, Patched receptor and Smoothened signal transducer. Although activation of Smoothened and its downstream signal transduction have been intensively studied, less is known about how Patched receptor is regulated, and particularly how this regulation contributes to appropriate Hedgehog signal transduction. Here we identified a novel role of Smurf E3 ligase in regulating Hedgehog signaling by controlling Patched ubiquitination and turnover. Moreover, we showed that Smurf-mediated Patched ubiquitination depends on Smo activity in wing discs. Mechanistically, we found that Smo interacts with Smurf and promotes it to mediate Patched ubiquitination by targeting the K1261 site in Ptc. The further mathematic modeling analysis reveals that a bidirectional control of activation of Smo involving Smurf and Patched is important for signal-receiving cells to precisely interpret external signals, thereby maintaining Hedgehog signaling reliability. Finally, our data revealed an evolutionarily conserved role of Smurf proteins in controlling Hh signaling by targeting Ptc during development.

Demographic and psychosocial correlates of sexual activity in older Chinese people.

AIMS AND OBJECTIVES: This study examines sexual activity and associated psychosocial factors in older Chinese people. BACKGROUND: Sexuality continues to play a pivotal role in our lives even as we grow old. There is, however, very limited research on the topic in older populations. DESIGN: Cross-sectional survey. METHODS: A representative sample of 688 older Chinese people (>60 years old) were individually interviewed on their demographic characteristics; their interest in, knowledge of and perceived control over sexual activities; and their engagement in sexual activity. RESULTS: The results show that 51·32% of men and 41·26% of women reported engaging in some form of sexual activity. Sexual intercourse and caressing were commonly reported. A multiple regression analysis also showed that a higher level of sexual activity was associated with being younger, living with a spouse, having a strong interest in sex, having sufficient knowledge of elder sexuality and a high perceived control over sex. Distinct predictive factors in each gender were observed. A high level of perceived control was associated with a higher level of sexual activity in males but not females. A series of binary logistic regression analyses were conducted to determine the demographic and psychosocial correlates of sexual activity. With the exception of the use of objects (sex toys), sexual activities were consistently associated with being younger; living with a spouse; and having high levels of interest, knowledge and perceived control. CONCLUSIONS: A sizable amount of older Chinese people engage in varying degrees of sexual activity, and most are still interested in sex. RELEVANCE TO CLINICAL PRACTICE: Frontline health professionals need to be aware of the growing needs for sex education in older persons, particular attention should be paid to discuss the limitations brought about by various chronic conditions associated with ageing and their relevance to elder sexuality.

[Prevention of Inonotus obliquus polysaccharides for high power microwave radiation induced testicular injury in rats: an experimental research].

OBJECTIVE: To investigate the effect of Inonotus obliquus polysaccharides on testicular injury induced by exposure to high power microwave (HPM) in rats. METHODS: A total of 30 male Wistar rats were randomly divided into 5 groups, i.e., the normal control group, the microwave radiation model group, the treatment group, the new microwave radiation model group, and the prevention group, 6 in each group. All rats, except those in the normal control group, were exposed to microwave at an average power density of 200 mW/cm2 for 6 min. Rats in the control group and the model group were administered with normal saline by gastrogavage, once a day. Rats in the treatment group and the prevention group were given with Inonotus obliquus polysaccharides by gastrogavage, 2 mL each time (400 mg/kg body weight), once a day. All rats were sacrificed on the 11th day.The sperm density and the rate of sperm deformity were determined. Pathological changes of testis were observed by light microscope and transmission electron microscope. RESULTS: Short-term HPM irradiation could significantly reduce the sperm density and increase the sperm deformity rate (P < 0.05). Meanwhile, obvious pathological changes of testes occurred. Compared with the two model groups, the sperm density increased and the sperm deformity rate decreased in the treatment group and the prevention group (P < 0.05). Under the light microscope, injuries of spermatogenic cells and stromal cells, as well as vascular dilatation and congestion were obviously alleviated in the treatment group and the prevention group. Mitochondrial swelling and endoplasmic reticulum expansion shown by ultrastructural observation were also significantly alleviated. Of them, injuries of spermatogenic cells and inflammation response were milder in the treatment group than in the prevention group. CONCLUSIONS: Inonotus obliquus polysaccharides had significant protective effect on microwave radiation induced testicular injury. Better effect was obtained by therapeutic medication than preventive medication.

Synergistic Tribological Effects of Ti3C2Tx MXene and ZDDP under Simple Grafting Strategies for Efficient Lubrication.

MXenes, a novel class of two-dimensional materials, possess exceptional physical and chemical properties, positioning them as promising candidates for lubricant additives. However, their potential is constrained by challenges in dispersion and stability, coupled with a paucity of research on interactions with additives in full-formula oils. In this study, hexadecylphosphonic acid (HDPA) is grafted onto Ti3C2Tx to formulate a polyalkylene glycol dispersion system. The findings reveal that the HDPA-modified Ti3C2Tx (HDPA-Ti3C2) is successfully synthesized, demonstrating superior dispersion stability and notable friction-reduction and antiwear properties. Notably, when combined with zinc dialkyl dithiophosphate (ZDDP), the HDPA-Ti3C2/ZDDP composite additive outperforms single additives in tribological performance, suggesting synergistic effects between them. This enhanced performance may be attributed to the formation of an amorphous polyphosphate tribofilm offering wear resistance, followed by the generation of a TiO2 tribofilm that further safeguards and repairs the worn surface, thereby enhancing the load-bearing capacity. Concurrently, the interlayer sliding mechanism of nanosheets, which substitutes the relative motion of the friction pair, reduces friction under boundary lubrication, ensuring prolonged effective lubrication. This work broadens the application prospects of Ti3C2Tx MXene for the design and development of commercial lubricating additives.