Dynamic nucleolar phase separation influenced by non-canonical function of LIN28A instructs pluripotent stem cell fate decisions.

LIN28A is important in somatic reprogramming and pluripotency regulation. Although previous studies addressed that LIN28A can repress let-7 microRNA maturation in the cytoplasm, few focused on its role within the nucleus. Here, we show that the nucleolus-localized LIN28A protein undergoes liquid-liquid phase separation (LLPS) in mouse embryonic stem cells (mESCs) and in vitro. The RNA binding domains (RBD) and intrinsically disordered regions (IDR) of LIN28A contribute to LIN28A and the other nucleolar proteins' phase-separated condensate establishment. S120A, S200A and R192G mutations in the IDR result in subcellular mislocalization of LIN28A and abnormal nucleolar phase separation. Moreover, we find that the naive-to-primed pluripotency state conversion and the reprogramming are associated with dynamic nucleolar remodeling, which depends on LIN28A's phase separation capacity, because the LIN28A IDR point mutations abolish its role in regulating nucleolus and in these cell fate decision processes, and an exogenous IDR rescues it. These findings shed light on the nucleolar function in pluripotent stem cell states and on a non-canonical RNA-independent role of LIN28A in phase separation and cell fate decisions.

Low-input lipidomics reveals lipid metabolism remodelling during early mammalian embryo development.

Lipids are indispensable for energy storage, membrane structure and cell signalling. However, dynamic changes in various categories of endogenous lipids in mammalian early embryonic development have not been systematically characterized. Here we comprehensively investigated the dynamic lipid landscape during mouse and human early embryo development. Lipid signatures of different developmental stages are distinct, particularly for the phospholipid classes. We highlight that the high degree of phospholipid unsaturation is a conserved feature as embryos develop to the blastocyst stage. Moreover, we show that lipid desaturases such as SCD1 are required for in vitro blastocyst development and blastocyst implantation. One of the mechanisms is through the regulation of unsaturated fatty-acid-mediated fluidity of the plasma membrane and apical proteins and the establishment of apical-basal polarity during development of the eight-cell embryo to the blastocyst. Overall, our study provides an invaluable resource about the remodelling of the endogenous lipidome in mammalian preimplantation embryo development and mechanistic insights into the regulation of embryogenesis and implantation by lipid unsaturation.

A second-generation M1-polarized CAR macrophage with antitumor efficacy.

Chimeric antigen receptor (CAR) T cell therapies have successfully treated hematological malignancies. Macrophages have also gained attention as an immunotherapy owing to their immunomodulatory capacity and ability to infiltrate solid tumors and phagocytize tumor cells. The first-generation CD3ζ-based CAR-macrophages could phagocytose tumor cells in an antigen-dependent manner. Here we engineered induced pluripotent stem cell-derived macrophages (iMACs) with toll-like receptor 4 intracellular toll/IL-1R (TIR) domain-containing CARs resulting in a markedly enhanced antitumor effect over first-generation CAR-macrophages. Moreover, the design of a tandem CD3ζ-TIR dual signaling CAR endows iMACs with both target engulfment capacity and antigen-dependent M1 polarization and M2 resistance in a nuclear factor kappa B (NF-κB)-dependent manner, as well as the capacity to modulate the tumor microenvironment. We also outline a mechanism of tumor cell elimination by CAR-induced efferocytosis against tumor cell apoptotic bodies. Taken together, we provide a second-generation CAR-iMAC with an ability for orthogonal phagocytosis and polarization and superior antitumor functions in treating solid tumors relative to first-generation CAR-macrophages.

Sexual dilemma among elderly men living with HIV/AIDS: a cross-sectional survey of the sexual needs and behaviours among HIV-positive men aged ≥50 years.

OBJECTIVES: This study aimed to explore the factors associated with the sexual needs and behaviours among elderly men living with HIV/AIDS in Chongqing, China. STUDY DESIGN: The study used a cross-sectional design. METHODS: Data were collected through face-to-face interviews using a structured questionnaire by convenience sampling from July to December 2021. The chi-squared test and logistic regression were conducted to identify factors associated with sexual needs and behaviours. RESULTS: In total, 63.70% of participants (493/774) reported having sexual needs. Individuals aged 50-59 years (odds ratio [OR] = 7.257, 95% confidence interval [CI]: 4.620, 11.401) and those who were married or cohabiting (OR = 1.595, 95% CI: 1.133, 2.246) reported high sexual needs. Among those with sexual needs, 69.17% (341/493) reported having sex in the past year. In total, 11.37% (40/341) and 9.68% (33/341) reported having commercial and casual sex, respectively. Participants aged 60-69 years (OR = 2.175, 95% CI: 1.128, 4.193), those who were married or cohabiting (OR = 3.371, 95% CI: 2.192, 5.184) and individuals who were employed (OR = 2.301, 95% CI: 1.339, 3.954) had higher odds of engaging in sexual behaviour, while participants with an awareness of AIDS-related knowledge (OR = 0.504, 95% CI: 0.307, 0.829) and those who had ≥1 year of antiretroviral therapy (ART) (ART 1-2 years: OR = 0.374, 95% CI: 0.176, 0.795; ART ≥3 years: OR = 0.429, 95% CI: 0.218, 0.846) had a lower likelihood of engaging in sexual behaviour. CONCLUSIONS: The sexual needs and behaviours of elderly men living with HIV/AIDS in Chongqing, China, remain at a high level. There was a discrepancy between sexual needs and behaviours among elderly men living with HIV/AIDS. Continuous dissemination of sexual health education and the promotion of condom use are necessary.

C-Myc-induced hypersialylation of small cell lung cancer facilitates pro-tumoral phenotypes of macrophages.

Immunosuppressive myeloid cell populations have been documented in small cell lung cancer (SCLC) subtypes, playing a key role in remolding the tumor microenvironment (TME). However, the cancer-associated transcriptional features of monocytes and tumor-associated macrophages (TAMs) in SCLC remain poorly understood. Herein, we analyzed the molecular features and functions of monocyte/macrophage subsets aiming to inhibit monocyte recruitment and pro-tumor behavior of macrophages. We observe that NEUROD1-high SCLC subtype (SCLC-N) exhibits subtype-specific hypersialylation induced by the unique target c-Myc (MYC) of NEUROD1. The hypersialylation can alter macrophage phenotypes and pro-tumor behavior by regulating the expression of the immune-inhibiting lectin receptors on monocyte-derived macrophages (MDMs) in SCLC-N. Inhibiting the aberrant sialic acid metabolic pathways in SCLC can significantly enhance the phagocytosis of macrophages. This study provides a comprehensive overview of the cancer-specific immune signature of monocytes and macrophages and reveals tumor-associated biomarkers as potential therapeutic targets for SCLC.

HIV-1 molecular transmission network and drug resistance in Chongqing, China, among men who have sex with men (2018-2021).

BACKGROUND: Over the past few years, HIV transmission among men who have sex with men (MSM) in China has increased significantly. Chongqing, located in the southwest of China, has the highest prevalence of HIV among MSM in the country. METHODS: Blood samples were taken from 894 MSM in Chongqing who had recently been diagnosed with HIV-1 infection and had not yet started getting treatment. In order to determine the distribution of HIV-1 subtypes, transmitted drug resistance, and assessments of molecularly transmitted clusters, we sequenced the Pol genes and employed them in phylogenetic analysis. The genetic distance between molecular clusters was 1.5%. To find potential contributing factors, logistic regression analyses were performed. RESULTS: Of the 894 HIV-1 pol sequences acquired from study participants, we discovered that CRF07_BC (73.6%) and CRF01_AE (19.6%) were the two most prevalent HIV-1 genotypes in Chongqing among MSM, accounting for 93.2% of all infections. In addition, CRF08_BC (1.1%), B subtype (1.0%), CRF55_01B (3.4%), and URF/Other subtypes (1.3%) were less frequently observed. Among MSM in Chongqing, transmitted drug resistance (TDR) was reported to be present at a rate of 5.6%. 48 clusters with 600 (67.1%, 600/894) sequences were found by analysis of the molecular transmission network. The distributions of people by age, sexual orientation, syphilis, and genotype were significantly differentially related to being in clusters, according to the multivariable logistic regression model. CONCLUSION: Despite the low overall prevalence of TDR, the significance of genotypic drug resistance monitoring needs to be emphasized. CRF07_BC and CRF01_AE were the two main genotypes that created intricate molecular transmission networks. In order to prevent the expansion of molecular networks and stop the virus's spread among MSM in Chongqing, more effective HIV intervention plans should be introduced.

Physiological response of anthocyanin synthesis to different light intensities in blueberry.

Fruit color is an important economic character of blueberry, determined by the amount of anthocyanin content. Anthocyanin synthesis within the blueberry fruits is significantly affected by light. To reveal the physiological response mechanism of anthocyanin synthesis in blueberry fruits in different light intensities, four light intensities (100% (CK), 75%, 50% and 25%) were set for the 'O'Neal' southern highbush blueberry as the experimental material in our study. The relationship between endogenous hormones content, associated enzyme activities, and variations with the anthocyanin content in blueberry fruits under various light intensities during the white fruit stage (S1), purple fruit stage (S2), and blue fruit stage (S3) were studied. The results showed that adequate light could significantly promote anthocyanin synthesis in blueberry fruits (P < 0.05). Blueberry fruits had an anthocyanin content that was 1.76~24.13 times higher under 100% light intensity than it was under non-full light intensity. Different light intensities significantly affected the content of endogenous hormones and the activity of associated enzymes in anthocyanin synthesis pathway (P < 0.05). Among them, the JA (jasmonic acid) content and PAL (phenylalanine ammonia lyase) activity of fruits under 100% light intensity were 2.49%~41.83% and 2.47%~48.48% higher than those under other light intensity, respectively. And a significant correlation was found between the variations in anthocyanin content in fruits and the content or activities of JA, ABA (abscisic acid), ETH (ethylene), GA3 (gibberellin 3), IAA (indoleacetic acid), PAL, CHI (chalcone isomerase), DFR (dihydroflavonol reductase) and UFGT (UDP-glucose: flavonoid 3-glucosyltransferase) (P < 0.05). It indicated that 100% light intensity significantly promoted anthocyanin synthesis in blueberry fruits by affecting endogenous hormones content and associated enzyme activities in the anthocyanin synthesis pathway. This study will lay a foundation for further research on the molecular mechanism of light intensity regulating anthocyanin synthesis in blueberry.

RAGE-TLR4 Crosstalk Is the Key Mechanism by Which High Glucose Enhances the Lipopolysaccharide-Induced Inflammatory Response in Primary Bovine Alveolar Macrophages.

The receptor of advanced glycation end products (RAGE) and Toll-like receptor 4 (TLR4) are important receptors for inflammatory responses induced by high glucose (HG) and lipopolysaccharide (LPS) and show crosstalk phenomena in inflammatory responses. However, it is unknown whether RAGE and TLR4 can influence each other's expression through a crosstalk mechanism and whether the RAGE-TLR4 crosstalk related to the molecular mechanism of HG enhances the LPS-induced inflammatory response. In this study, the implications of LPS with multiple concentrations (0, 1, 5, and 10 µg/mL) at various treatment times (0, 3, 6, 12, and 24 h) in primary bovine alveolar macrophages (BAMs) were explored. The results showed that a 5 µg/mL LPS treatment at 12 h had the most significant increment on the pro-inflammatory cytokine interleukin 1ß (IL-1ß), IL-6, and tumor necrosis factor (TNF)-α levels in BAMs (p < 0.05) and that the levels of TLR4, RAGE, MyD88, and NF-κB p65 mRNA and protein expression were upregulated (p < 0.05). Then, the effect of LPS (5 µg/mL) and HG (25.5 mM) co-treatment in BAMs was explored. The results further showed that HG significantly enhanced the release of IL-1ß, IL-6, and TNF-α caused by LPS in the supernatant (p < 0.01) and significantly increased the levels of RAGE, TLR4, MyD88, and NF-κB p65 mRNA and protein expression (p < 0.01). Pretreatment with FPS-ZM1 and TAK-242, the inhibitors of RAGE and TLR4, significantly alleviated the HG + LPS-induced increment of RAGE, TLR4, MyD88, and NF-κB p65 mRNA and protein expression in the presence of HG and LPS (p < 0.01). This study showed that RAGE and TLR4 affect each other's expression through crosstalk during the combined usage of HG and LPS and synergistically activate the MyD88/NF-κB signaling pathway to promote the release of pro-inflammatory cytokines in BAMs.

Personal glucose meters coupled with signal amplification technologies for quantitative detection of non-glucose targets: Recent progress and challenges in food safety hazards analysis.

Ensuring food safety is paramount worldwide. Developing effective detection methods to ensure food safety can be challenging owing to trace hazards, long detection time, and resource-poor sites, in addition to the matrix effects of food. Personal glucose meter (PGM), a classic point-of-care testing device, possesses unique application advantages, demonstrating promise in food safety. Currently, many studies have used PGM-based biosensors and signal amplification technologies to achieve sensitive and specific detection of food hazards. Signal amplification technologies have the potential to greatly improve the analytical performance and integration of PGMs with biosensors, which is crucial for solving the challenges associated with the use of PGMs for food safety analysis. This review introduces the basic detection principle of a PGM-based sensing strategy, which consists of three key factors: target recognition, signal transduction, and signal output. Representative studies of existing PGM-based sensing strategies combined with various signal amplification technologies (nanomaterial-loaded multienzyme labeling, nucleic acid reaction, DNAzyme catalysis, responsive nanomaterial encapsulation, and others) in the field of food safety detection are reviewed. Future perspectives and potential opportunities and challenges associated with PGMs in the field of food safety are discussed. Despite the need for complex sample preparation and the lack of standardization in the field, using PGMs in combination with signal amplification technology shows promise as a rapid and cost-effective method for food safety hazard analysis.

Glucometer-based biosensor for the determination of ractopamine in animal-derived foods using rolling circle amplification.

Screening for persistent organic pollutants (POPs) in food is a complex and challenging process, as POPs can be present in very low levels and can be difficult to detect. Herein, we developed an ultrasensitive biosensor based on a rolling circle amplification (RCA) platform using a glucometer to determine POP. The biosensor was constructed using gold nanoparticle probes modified with antibodies and dozens of primers, magnetic microparticle probes conjugated with haptens, and targets. After competition, RCA reactions are triggered, numerous RCA products hybridize with the ssDNA-invertase, and the target is successfully transformed into glucose. Using ractopamine as a model analyte, this strategy obtained a linear detection range of 0.038-5.00 ng mL-1 and a detection limit of 0.0158 ng mL-1, which was preliminarily verified by screening in real samples. Compared with conventional immunoassays, this biosensor utilizes the high efficiency of RCA and the portable properties of a glucometer, which effectively improves the sensitivity and simplifies the procedures using magnetic separation technology. Moreover, it has been successfully applied to ractopamine determination in animal-derived foods, revealing its potential as a promising tool for POP screening.

Bimetal-MOF and bacterial cellulose-derived three-dimensional N-doped carbon sheets loaded Co/CoFe nanoparticles wrapped graphite carbon supported on porous carbon nanofibers: An efficient multifunctional electrocatalyst for Zn-air batteries and overall water splitting.

In this work, a three-dimensional (3D) multifunctional Co/CoFeNC@N-CNF electrocatalyst was first synthesized by the pyrolysis of a CoFe bimetal-centred metal-organic framework (MOF) and bacterial cellulose (BC). The initial potential and half-wave potential of Co/CoFeNC@N-CNF can reach 0.99 V and 0.8 V. Low overpotentials of 320 mV and 155 mV are purely required for the oxygen evolution reaction (OER) and hydrogen evolution reaction (HER) at a current density of 10 mA cm-2, respectively. The electrochemical performance of Co/CoFeNC@N-CNF exceeds most bimetal-MOF-derived electrocatalysts reported to date. The superior electrochemical performance is mainly due to abundant active sites, high-efficiency electrochemical performance, and high electron transport efficiency. In addition, the theoretical calculation results show that the synergistic effect of the CoFe bimetal can optimize the adsorption energy for intermediates of the oxygen reduction reaction (ORR), OER and HER. Furthermore, we assembled a mold and solid Zn-air battery using the catalyst as an air cathode catalyst, demonstrating the maximum power densities of 292 mW cm-2 and 178 mW cm-2. The 3D structure electrocatalysts derived from the MOF and bacterial cellulose provide an innovative and instructive approach for the design of diverse energy nanomaterials.

Depression and Associated Factors Among Men Living with HIV/AIDS Aged 50 Years and Over in Chongqing, China.

Purpose: This study aimed to assess the proportion of depression and its associated factors among men living with HIV/AIDS and aged 50 and over in Chongqing, China. Methods: This cross-sectional study included 774 men diagnosed as HIV/AIDS and aged 50 and over in four regions. Data were collected through face-to-face interviews with a structured questionnaire. Multivariate logistic regression analysis was performed to identify factors associated with depression. A two-tailed P-value less than 0.05 was considered as statistical significance. Results: A total of 293 participants (37.9%) reported depressive symptoms. Results from multivariate logistic regression suggested that individuals aged ≥70 (AOR = 1.99, 95% CI: 1.28-3.08) and those living in rural areas (AOR = 2.79, 95% CI: 1.96-3.97) were associated with higher odds of depression, while those being employed (AOR = 0.50, 95% CI: 0.32-0.80) and with monthly income >3000 CNY (AOR = 0.47, 95% CI: 0.30-3.74) had lower odds of depression. Conclusion: Our findings indicate a high prevalence of depression among older men living with HIV/AIDS in Chongqing, China. Age, residence, employment, and income have been identified as potential risk factors of depressive symptoms. Mental-health intervention initiatives should be tailored to target adults with a higher risk of depression.

[Simultaneous determination of 40 plant growth regulators, fungicides, insecticides, and antibiotics in bean sprouts by QuEChERS-high performance liquid chromatography-tandem mass spectrometry].

Chromatography combined with mass spectrometry is the most commonly used detection technology, and it offers the advantages of high sensitivity and high selectivity. The quick, easy, inexpensive, effective, rugged, and safe (QuEChERS) method is low-cost, effective, and time efficient. The application of the QuEChERS has now been extended to the analysis of contaminants in food samples. The aim of the study was to identify different concentration levels of multiple harmful drug residues in bean sprouts. In this study, QuEChERS coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was established for the simultaneous determination of 40 plant growth regulators, fungicides, insecticides, and antibiotics in bean sprouts. In the HPLC-MS/MS experiment, gibberellic acid, 4-fluorophenoxyacetic acid, chloramphenicol, N6-(δ2-isopentenyl)-adenine, 6-benzylaminopurine, 4-chlorophenoxyacetic acid, and 2,4-dichlorophenoxyacetic acid (2,4-D) were analyzed by MS/MS with negative electrospray ionization (ESI-). The other 33 target analytes (chlormequat, ronidazole, metronidazole, pymetrozine, dimetridazole, methomyl, carbendazim, enoxacin, levofloxacin, pefloxacin mesylate, norfloxacin, ciprofloxacin, enrofloxacin, thiabendazole, lomefloxacin, chlorpyrifos, sarafloxacin, imidacloprid, etc.) were analyzed by MS/MS with positive electrospray ionization (ESI+). Sensitive MS conditions were realized by optimizing the instrumental parameters such as the desolvent temperature, collision energy, spraying needle position, precursor ions, and product ions. Then, the optimal pretreatment method was determined by comparing the recovery rates of the 40 drugs obtained with different extraction solvents (methanol, acetonitrile, acetonitrile containing 0.1% ammonia, acetonitrile with 1% acetic acid), different extraction methods (ultrasonic extraction, shaking extraction), and purification with primary secondary amine (PSA) and C18. In this study, the bean sprouts samples were extracted twice by 10 mL acetonitrile with 1% acetic acid, and extracted under ultrasonic conditions. Then, the extracting solution was only cleaned with 100 mg C18. The chromatographic separation of the 40 compounds was accomplished on a Waters ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 µm) with gradient elution. Methanol and 0.01% formic acid aqueous solution were used as the mobile phases. The 40 compounds were analyzed in the multiple reaction monitoring (MRM) mode. The matrix matching external standard method was used for quantitative determination. The results showed that the 40 compounds could be analyzed within 15 min. Under the optimized conditions, the calibration curves showed good linearities for the 40 compounds, and the coefficients of determination (r2) were greater than 0.99 in the range of 2-200 µg/L. The limits of detection (LODs) and limits of quantification (LOQs) were in the range of 0.1-3 µg/kg and 0.3-9 µg/kg, respectively. Using negative bean sprouts as the substrates, the recovery tests were carried out at three spiked levels of 5, 10, and 50 µg/kg. The average recoveries of the 40 drugs were 78.5% to 115.3%, and the corresponding relative standard deviations (RSDs) were 1.3% to 9.7% (n=6). This method was successfully applied to the analysis of the 40 drug residues in 21 batches of local bean sprouts in Handan city. The results revealed the presence of extensive drug residues in the bean sprouts. The 26 batches were detected to varying degrees, among which 4-chlorophenoxyacetic acid, carbendazim, 6-benzyladenine, 2,4-D, enrofloxacin, and metronidazole were detected at high rates. The detection rates of 4-chlorophenoxyacetic acid, 6-benzyladenine, carbendazim, 2,4-D, gibberellic acid, and enrofloxacin were 28.6%, 19.0%, 9.5%, 9.5%, 4.8%, and 4.8%, respectively. The contents ranged from 37.5-352.4, 32.4-273.1, 28.8-38.7, 316.1-20.2, 19.9 and 13.6 µg/kg, respectively. Given its advantages of simplicity, rapidness, and high sensitivity, the developed method can be used for the rapid and accurate determination of trace levels of the 40 drug residues in large quantities of bean sprouts.

Macrophage-Based Combination Therapies as a New Strategy for Cancer Immunotherapy.

BACKGROUND: Cells of the immune system can inhibit tumor growth and progression; however, immune cells can also promote tumor cell growth, survival, and angiogenesis as a result of the immunosuppressive microenvironments. In the last decade, a growing number of new therapeutic strategies focused on reversing the immunosuppressive status of tumor microenvironments (TMEs), to reprogram the TME to be normal, and to further activate the antitumor functions of immune cells. Most of the "hot tumors" are encompassed with M2 macrophages promoting tumor growth, and the accumulation of M2 macrophages into tumor islets leads to poor prognosis in a wide variety of tumors. SUMMARY: Therefore, how to uncover more immunosuppressive signals and to reverse the M2 tumor-associated macrophages (TAMs) to M1-type macrophages is essential for reversing the immunosuppressive state. Except for reeducation of TAMs in the cancer immunotherapy, macrophages as central effectors and regulators of the innate immune system have the capacity of phagocytosis and immune modulation in macrophage-based cell therapies. KEY MESSAGES: We review the current macrophage-based cell therapies that use genetic engineering to augment macrophage functionalities with antitumor activity for the application of novel genetically engineered immune cell therapeutics. A combination of TAM reeducation and macrophage-based cell strategy may bring us closer to achieving the original goals of curing cancer. In this review, we describe the characteristics, immune status, and tumor immunotherapy strategies of macrophages to provide clues and evidences for future macrophage-based immune cell therapies.

LIN28 coordinately promotes nucleolar/ribosomal functions and represses the 2C-like transcriptional program in pluripotent stem cells.

LIN28 is an RNA binding protein with important roles in early embryo development, stem cell differentiation/reprogramming, tumorigenesis and metabolism. Previous studies have focused mainly on its role in the cytosol where it interacts with Let-7 microRNA precursors or mRNAs, and few have addressed LIN28's role within the nucleus. Here, we show that LIN28 displays dynamic temporal and spatial expression during murine embryo development. Maternal LIN28 expression drops upon exit from the 2-cell stage, and zygotic LIN28 protein is induced at the forming nucleolus during 4-cell to blastocyst stage development, to become dominantly expressed in the cytosol after implantation. In cultured pluripotent stem cells (PSCs), loss of LIN28 led to nucleolar stress and activation of a 2-cell/4-cell-like transcriptional program characterized by the expression of endogenous retrovirus genes. Mechanistically, LIN28 binds to small nucleolar RNAs and rRNA to maintain nucleolar integrity, and its loss leads to nucleolar phase separation defects, ribosomal stress and activation of P53 which in turn binds to and activates 2C transcription factor Dux. LIN28 also resides in a complex containing the nucleolar factor Nucleolin (NCL) and the transcriptional repressor TRIM28, and LIN28 loss leads to reduced occupancy of the NCL/TRIM28 complex on the Dux and rDNA loci, and thus de-repressed Dux and reduced rRNA expression. Lin28 knockout cells with nucleolar stress are more likely to assume a slowly cycling, translationally inert and anabolically inactive state, which is a part of previously unappreciated 2C-like transcriptional program. These findings elucidate novel roles for nucleolar LIN28 in PSCs, and a new mechanism linking 2C program and nucleolar functions in PSCs and early embryo development.

rRNA biogenesis regulates mouse 2C-like state by 3D structure reorganization of peri-nucleolar heterochromatin.

The nucleolus is the organelle for ribosome biogenesis and sensing various types of stress. However, its role in regulating stem cell fate remains unclear. Here, we present evidence that nucleolar stress induced by interfering rRNA biogenesis can drive the 2-cell stage embryo-like (2C-like) program and induce an expanded 2C-like cell population in mouse embryonic stem (mES) cells. Mechanistically, nucleolar integrity maintains normal liquid-liquid phase separation (LLPS) of the nucleolus and the formation of peri-nucleolar heterochromatin (PNH). Upon defects in rRNA biogenesis, the natural state of nucleolus LLPS is disrupted, causing dissociation of the NCL/TRIM28 complex from PNH and changes in epigenetic state and reorganization of the 3D structure of PNH, which leads to release of Dux, a 2C program transcription factor, from PNH to activate a 2C-like program. Correspondingly, embryos with rRNA biogenesis defect are unable to develop from 2-cell (2C) to 4-cell embryos, with delayed repression of 2C/ERV genes and a transcriptome skewed toward earlier cleavage embryo signatures. Our results highlight that rRNA-mediated nucleolar integrity and 3D structure reshaping of the PNH compartment regulates the fate transition of mES cells to 2C-like cells, and that rRNA biogenesis is a critical regulator during the 2-cell to 4-cell transition of murine pre-implantation embryo development.

Metagenomics Reveals That Intravenous Injection of Beta-Hydroxybutyric Acid (BHBA) Disturbs the Nasopharynx Microflora and Increases the Risk of Respiratory Diseases.

It is widely accepted that maintenance of microbial diversity is essential for the health of the respiratory tract; however, there are limited reports on the correlation between starvation and respiratory tract microbial diversity. In the present study, saline/ß-hydroxybutyric acid (BHBA) intravenous injection after dietary restriction was used to imitate different degrees of starvation. A total of 13 healthy male yaks were imposed to different dietary restrictions and intravenous injections, and their nasopharyngeal microbiota profiles were obtained by metagenomic shotgun sequencing. In healthy yaks, the main dominant phyla were Proteobacteria (33.0%), Firmicutes (22.6%), Bacteroidetes (17.2%), and Actinobacteria (13.2%); the most dominated species was Clostridium botulinum (10.8%). It was found that 9 days of dietary restriction and 2 days of BHBA injection (imitating severe starvation) significantly decreased the microbial diversity and disturbed its structure and functional composition, which increased the risk of respiratory diseases. This study also implied that oral bacteria played an important role in maintaining nasopharynx microbial homeostasis. In this study, the correlation between starvation and nasopharynx microbial diversity and its potential mechanism was investigated for the first time, providing new ideas for the prevention of respiratory diseases.

Comparative Evaluation of Integrated Waste Heat Utilization Systems for Coal-Fired Power Plants Based on In-Depth Boiler-Turbine Integration and Organic Rankine Cycle.

To maximize the system-level heat integration, three retrofit concepts of waste heat recovery via organic Rankine cycle (ORC), in-depth boiler-turbine integration, and coupling of both are proposed, analyzed and comprehensively compared in terms of thermodynamic and economic performances. For thermodynamic analysis, exergy analysis is employed with grand composite curves illustrated to identify how the systems are fundamentally and quantitatively improved, and to highlight key processes for system improvement. For economic analysis, annual revenue and investment payback period are calculated based on the estimation of capital investment of each component to identify the economic feasibility and competitiveness of each retrofit concept proposed. The results show that the in-depth boiler-turbine integration achieves a better temperature match of heat flows involved for different fluids and multi-stage air preheating, thus a significant improvement of power output (23.99 MW), which is much larger than that of the system with only ORC (6.49 MW). This is mainly due to the limitation of the ultra-low temperature (from 135 to 75 °C) heat available from the flue gas for ORC. The thermodynamic improvement is mostly contributed by the reduction of exergy destruction within the boiler subsystem, which is eventually converted to mechanical power; while the exergy destruction within the turbine system is almost not changed for the three concepts. The selection of ORC working fluids is performed to maximize the power output. Due to the low-grade heat source, the cycle with R11 offers the largest additional net power generation but is not significantly better than the other preselected working fluids. Economically, the in-depth boiler-turbine integration is the most economic completive solution with a payback period of only 0.78 year. The ORC concept is less attractive for a sole application due to a long payback time (2.26 years). However, by coupling both concepts, a net power output of 26.51 MW and a payback time of almost one year are achieved, which may promote the large-scale production and deployment of ORC with a cost reduction and competitiveness enhancement.

Changes in the smoking behavior of pregnant women andtheir family members during pregnancy: A cross-sectional study in China.

INTRODUCTION: Studies regarding the changes in smoking behavior of pregnant women have been mostly conducted in high income countries but rarely in China. This study thus focused on investigating the changes in smoking behavior among pregnant women and their family members in China, both before and during pregnancy. METHODS: A cross-sectional study was carried out at nine Women and Children's Hospitals in Shanghai, China, in 2014. A total of 2831 gestational households were recruited. The chi-squared test, paired sample t-test and logistical regression analysis were used during statistical analysis. RESULTS: The prevalence of smoking for all household members significantly declined during pregnancy: 76.2% of the pregnant women, 19.2% of their husbands and 14.0% of other family members quit smoking entirely. The average daily cigarette consumption rate decreased from 5.9 to 0.9 among pregnant women, 11.4 to 9.5 for husbands, and 11.4 to 9.5 for other family members (paired sample t-test, p<0.001). The likelihood that husbands continued smoking during pregnancy was significantly lower for those with a shorter history of smoking, had a lower daily cigarette consumption rate and a household registration in Shanghai. CONCLUSIONS: The prevalence of smoking among pregnant women, husbands and other family members significantly declines during pregnancy. Thus, pregnancy is most likely a key period in which to provide families with health education regarding the effects of smoking, both during the pregnancy period and in general. Intervention programs designed to reduce smoking among husbands during pregnancy should focus on those with a long history of smoking, a high rate of daily cigarette consumption, and those with household registration not in Shanghai.