Weak-Water-Coordination Electrolyte to Stabilize Zinc Anode Interface for Aqueous Zinc Ion Batteries.

The performance of zinc-ion batteries is severely hindered by the uncontrolled growth of dendrites and the severe side reactions on the zinc anode interface. To address these challenges, a weak-water-coordination electrolyte is realized in a peptone-ZnSO4 -based electrolyte to simultaneously regulate the solvation structure and the interfacial environment. The peptone molecules have stronger interaction with Zn2+ ions than with water molecules, making them more prone to coordinate with Zn2+ ions and then reducing the active water in the solvated sheath. Meantime, the peptone molecules selectively adsorb on the Zn metal surface, and then are reduced to form a stable solid-electrolyte interface layer that can facilitate uniform and dense Zn deposition to inhabit the dendritic growth. Consequently, the Zn||Zn symmetric cell can exhibit exceptional cycling performance over 3200 h at 1.0 mA cm-2 /1.0 mAh cm-2 in the peptone-ZnSO4 -based electrolyte. Moreover, when coupled with a Na2 V6 O16 ·3H2 O cathode, the cell exhibits a long lifespan of 3000 cycles and maintains a high capacity retention rate of 84.3% at 5.0 A g-1 . This study presents an effective approach for enabling simultaneous regulation of the solvation structure and interfacial environment to design a highly reversible Zn anode.

A Fluorinated Solid-state-electrolyte Interface Layer Guiding Fast Zinc-ion Oriented Deposition in Aqueous Zinc-ion Batteries.

Aqueous zinc ion batteries are gaining popularity due to their high energy density and environmental friendliness. However, random deposition of zinc ions on the anode and sluggish migration of zinc ions on the interface would lead to the growth of zinc dendrites and poor cycling performance. To address these challenges, we developed a fluorinated solid-state-electrolyte interface layer composed of Ca5 (PO4 )3 F/Zn3 (PO4 )2 via an in situ ion exchange strategy to guide zinc-ion oriented deposition and fast zinc ion migration on the anode during cycling. The introduction of Ca5 (PO4 )3 F (FAP) can increase the nucleation sites of zinc ions and guide the oriented deposition of zinc ions along the (002) crystal plane, while the in situ formation of Zn3 (PO4 )2 during cycling can accelerate the migration of zinc ions. Benefited from our design, the assembled Zn//V2 O5 ⋅ H2 O batteries based on FAP-protected Zn anode (FAP-Zn) achieve a higher capacity retention of 84 % (220 mAh g-1 ) than that of bare-Zn based batteries, which have a capacity retention of 23 % (97 mAh g-1 ) at 3.0 A g-1 after 800 cycles. This work provides a new solution for the rational design and development of the solid-state electrolyte interface layer to achieve high-performance zinc-ion batteries.

Thermosipho ferrireducens sp.nov., an anaerobic thermophilic iron(III)-reducing bacterium isolated from a deep-sea hydrothermal sulfide deposits.

A thermophilic, anaerobic, iron-reducing bacterium strain JL129W03T (=KCTC 15905T=MCCC 1A14213T) was isolated from a sulfide sample collected from the Daxi hydrothermal field (60.5° E, 6.4° N, 2919 m depth) on the Carlsberg Ridge, northwest Indian Ocean. Cells grew at 55-75 °C(optimum, 70 °C), at pH 6.0-9.0 (optimum, pH 6.0-7.0) and at NaCl concentrations of 1.5-4.5 % (w/v; optimum 3.0 %). Under optimal growth conditions, the generation time was around 85 min. The isolate was an obligate chemoorganoheterotroph, utilizing complex organic compounds, carbohydrates, organic acids and one amino acid. It was anaerobic and facultatively dependent on elemental sulphur and various forms of Fe(III) as an electron acceptor: insoluble forms and soluble forms. It did not reduce sulfite, sulphate, thiosulfate or nitrate. The G+C content of its genomic DNA was 34.0 mol%. Phylogenetic 16S rRNA gene sequence analyses revealed that its closest relative was Thermosipho atlanticus DV1140T with 95.81 % 16S rRNA sequence similarity. On the basis of physiological distinctness and phylogenetic distance, the isolate is considered to represent a novel species of the genus Thermosipho, for which the name Thermosipho ferrireducens sp. nov. is proposed. The type strain is strain JL129W03T (=KCTC 15905T;=MCCC 1A14213T).

Correlation of Homocysteine, AHSG, CRP with Insulin Resistance, 25-(OH)2-VitD, Blood Lipids in Gestational Diabetes Patients.

BACKGROUND: To investigate the correlations of serum homocysteine (Hcy), α2-Heremans-Schmid glycoprotein (AHSG), and C-reactive protein (CRP) with insulin resistance (IR), 25-hydroxyvitamin D (25-OH-VD), and blood lipids in patients with gestational diabetes mellitus (GDM) by detecting their levels. METHODS: A total of 72 GDM patients (GDM group) and 72 healthy pregnant women (control group) delivered in our hospital from February 2017 to January 2019 were randomly selected. The basic data, somatological parameters [height, weight, body mass index (BMI), waist circumference, hip circumference, waist-to-hip ratio (WHR), blood pressure, and body fat content], and biochemical indexes (glucose metabolism indexes, lipid metabolism indexes, Hcy, AHSG, CRP, and 25-OH-VD) were compared between the two groups. Additionally, Pearson's correlation analysis was employed to analyze the correlations among indicators. RESULTS: In comparison with the control group, the GDM group had a higher average rate of family history of DM (p < 0.05), larger waist circumference and WHR, and higher body fat content (p < 0.05). Besides, the fasting plasma glucose (FPG), 1-hour plasma glucose (1hPG) and 2-hour plasma glucose (2hPG), glycosylated hemoglobin (HbA1c), fasting insulin (FINS), homeostasis model assessment (HOMA)-IR, triglyceride (TG), total cholesterol (TC), and low density lipoprotein cholesterol (LDL-C) were higher in the GDM group than those in the control group (p < 0.05), while the high density lipoprotein cholesterol (HDL-C) was lower in the GDM group than that in the control group (p < 0.05). Compared with those in the control group, the serum Hcy, AHSG, and CRP levels rose, while the serum 25-OH-VD level declined in the GDM group (p < 0.05). The results of Pearson's correlation analysis revealed that HOMA-IR had positive correlations with FPG, FINS, TC, TG, Hcy, AHSG, and CRP (r = 0.591, 0.825, 0.312, 0.234, 0.458, 0.647, 0.487, p < 0.05) and negative correlation with 25-OH-VD (r = -0.323, p < 0.05). CRP was positively correlated with HOMA-IR, TC, and AHSG (r = 0.485, 0.331, 0.226, p < 0.05), negatively associated with 25-OH-VD (r = -0.443, p < 0.05), and had no correlation to TG and Hcy (r = 0.019, 0.058, p > 0.05). AHSG displayed positive correlations with HOMA-IR, TC, TG, and CRP (r = 0.647, 0.321, 0.314, 0.226, p < 0.05) and no association with Hcy and 25-OH-VD (r = 0.058, -0.034, p > 0.05). CONCLUSIONS: GDM patients have increased serum Hcy, AHSG, and CRP levels and a decreased serum 25-OH-VD level, indicating that serum Hcy, AHSG, CRP, and 25-OH-VD are correlated with glucose and lipid metabolism disorders in GDM patients.

Interleukin-36 receptor antagonist attenuates atherosclerosis development by inhibiting NLRP3 inflammasome.

Atherosclerosis is characterized, as an inflammatory disorder in the circulatory system, with increasing tendency toward mortality and morbidity. Thus, developing novel therapeutic targeting inflammation is necessary. Here, we investigated the effects of interleukin-36 receptor antagonist (IL-36RN), a newly identified anti-inflammatory factor, on atherosclerosis. The regulation of NLRP3 inflammasome by IL-36RN was determined in vitro in macrophage cells after oxidized low-density lipoprotein (ox-LDL) stimulation. The IL-1ß and caspase-1 p10 secretion were assessed by enzyme-linked immunosorbent assay and western blot analysis. Finally, the IL-36RN/NLRP3 inflammasome pathway was confirmed in apolipoprotein E-deficient mice. IL-36RN suppressed the expression of NLRP3, the secretion of IL-1ß, and caspase-1 p10 in vitro, while IL-36 pathway stimulation activated the NLRP3 inflammasome, which was inhibited by IL-36RN. In the mouse model of atherosclerosis, IL-36RN delivered by the lentivirus vector inhibited the development of atherosclerosis, and the atheroprotective effects of IL-36RN were attenuated by IL-36 pathway stimulation. Furthermore, the regulation of NLRP3 inflammasome by IL-36RN was also confirmed in vivo. We demonstrated here that IL-36RN exerted atheroprotective functions through IL-36RN/NLRP3 inflammasome pathway.

Identifying Associations Between Influenza Vaccination Status and Access, Beliefs, and Sociodemographic Factors Among the Uninsured Population in Suffolk County, NY.

Seasonal influenza continues to impose a substantial healthcare and economic burden in the United States each year. This study assessed influenza vaccination rates among the uninsured population, determined the need for free flu vaccines, and analyzed associations of vaccination status with access, beliefs, and sociodemographic factors. Stony Brook Outreach and Medical Education (SB HOME) free clinic offers free flu vaccinations to uninsured patients over the age of 18 living in Suffolk County, New York. All patients visiting SB HOME between October 2018 and February 2019 were offered a 10-min anonymous survey assessing demographic characteristics, self-evaluation of health status, vaccination status during the 2017-2018 and 2018-2019 flu seasons, willingness to obtain a free flu vaccine, and beliefs and barriers surrounding flu vaccines. Analysis consisted of descriptive statistics, paired t tests, and χ2 tests using a significance threshold of p < 0.05. A total of 102 participants were surveyed, 80% of whom identified as Hispanic/Latino. The majority of participants expressed positive attitudes towards the influenza vaccine. Despite these positive attitudes, 72% reported not receiving the influenza vaccination during the 2017-2018 flu season, the commonly cited reasons being fear of getting sick from the flu vaccine, high cost, and not knowing where to get the vaccine. During the 2018-2019 flu season 60% of participants elected to receive a free flu shot at SB HOME, and participants with positive attitudes towards flu vaccines were more likely to accept the vaccine when offered at no cost. Fear of getting sick from the flu vaccine remained the most common reason for participants declining a free flu vaccine. Only 17% of participants would seek vaccination at the market price of $30, compared to the 76% who would if the vaccine was offered for free. Our study highlights the need for convenient and affordable access to the flu vaccine among uninsured populations, who despite recognizing the benefits of receiving the flu vaccine are unable to overcome certain barriers. Elucidating and targeting barriers specific to uninsured populations at the local community level may improve public health strategies centered on increasing vaccination rates.

Dynamic microglial modulation of spatial learning and social behavior.

Microglia are active players in inflammation, but also have important supporting roles in CNS maintenance and function, including modulation of neuronal activity. We previously observed an increase in the frequency of excitatory postsynaptic current in organotypic brain slices after depletion of microglia using clodronate. Here, we describe that local hippocampal depletion of microglia by clodronate alters performance in tests of spatial memory and sociability. Global depletion of microglia by high-dose oral administration of a Csf1R inhibitor transiently altered spatial memory but produced no change in sociability behavior. Microglia depletion and behavior effects were both reversible, consistent with a dynamic role for microglia in the regulation of such behaviors.

Visible-light-promoted syntheses of ß-keto sulfones from alkynes and sulfonylhydrazides.

A variety of functionalized ß-keto sulfones were smoothly prepared through oxysulfonylation of commercially available alkynes with sulfonylhydrazides under the synergistic interactions of visible light irradiation, Ru(bpy)3Cl2 photocatalyst, oxygen, KI, and NaOAc basic additive under very mild reaction conditions.

Role of ERp46 in ß-cell lipoapoptosis through endoplasmic reticulum stress pathway as well as the protective effect of exendin-4.

Endoplasmic reticulum (ER) stress is considered as a key factor in free fatty acid (FFA)-induced apoptosis. ERp46, a new member of the thioredoxin family, is highly expressed in pancreatic ß-cells and plays an important role in glucose toxicity. In this study we examined the potential role of ERp46 in palmitic acid (PA)-induced cell apoptosis and the protective role of exendin-4, a long-acting agonist of the hormone glucagon-like peptide-1 (GLP-1) receptor. The glucose-sensitive mouse ß-pancreatic cell line, ßTC6, was used to investigate the mechanisms of PA-induced apoptosis. Our results showed that ERp46 expression was reduced in a dose- and time-dependent manner after PA treatment. Furthermore, inhibition of ERp46 expression by small interfering (si)RNA-mediated silencing enhanced the ER stress response via three separate pathways and increased ßTC6 cell apoptosis rates. Moreover, exendin-4 reduced the ER stress response and levels of apoptosis in NC transfected cells after PA treatment, but not in cells transfected with ERp46siRNA. In conclusion, ERp46 plays a protective role in PA-induced cell apoptosis by decreasing the ER stress response and might be a novel target for anti-diabetic drugs. Exendin-4 might protect against ßTC6 cell lipoapoptosis in part by activating ERp46 signaling pathway.

Effect of PANDER in ßTC6-cell lipoapoptosis and the protective role of exendin-4.

Chronic exposure to high concentrations of saturated fatty acids, such as palmitic acid (PA), leads to apoptosis of pancreatic ß-cells through the activation of the c-Jun N-terminal kinase (JNK) signaling pathway. This study of ß-cell lipoapoptosis was designed to investigate the roles of pancreatic-derived factor (PANDER), a pro-apoptosis cytokine-like peptide, and exendin-4, a long-acting agonist of the hormone glucagon-like peptide-1 (GLP-1) receptor and anti-apoptosis factor. The glucose-sensitive mouse ß-pancreatic cell line, ßTC6, was used to investigate the mechanisms of PA-induced apoptosis. Twenty-four hours of PA exposure led to increased PANDER expression in a dose- and time-dependent manner, and significantly increased phosphorylation of JNK. Treatment with the JNK-specific inhibitor SP600125 reduced the PA-induced PANDER expression. After the 24h of PA exposure, cells also underwent marked apoptosis and showed increased activation of the apoptosis protease, caspase-3. The small interfering (si)RNA-mediated silencing of PANDER gene expression significantly reduced both of these effects. When PA-treated ßTC6 cells were exposed to exogenous exendin-4, JNK activation was inhibited, PANDER expression was decreased, and the numbers of apoptotic cells were reduced. Collectively, these results demonstrated that the JNK-mediated signaling mechanism of PA-induced ß-cell apoptosis involves up-regulated expression of PANDER and activation of caspase-3. Exendin-4 may protect against lipoapoptosis by interfering with the JNK-PANDER pathway.

Functional Zoning and Path Selection of Land Comprehensive Consolidation Based on Grey Constellation Clustering: A Case Study of Dongying City, China.

The functional zoning of land comprehensive consolidation and the selection of consolidation paths are the key content of the current land and space planning, and it is also an important measure to achieve regional sustainable development. At present, the research system on land comprehensive consolidation is not yet mature. The previous research area is relatively small and not representative, and an effective method system has not been formed. Research on the selection of functional zoning and a consolidation path is also relatively scarce. There is an urgent need to construct the theory and method system of land comprehensive consolidation functional zoning and consolidation path selection. Taking Dongying City in China as an example, this paper constructs a zoning index system from four aspects including natural conditions, location advantages, social economy and land use. The entropy method is used to determine the weights, and GIS spatial data visualization is used to analyze the spatial distribution characteristics of the index system. Based on the analysis, the grey constellation clustering method is used to divide the study area into four types of land comprehensive consolidation functional areas, and the results of functional zoning are adjusted according to the ranking of comprehensive index values, the principle of maximum similarity, and the continuity of natural space. The research results show that: ① There are 10 functional zonings for urban development and ecological protection land consolidation, and the main consolidation path should be the ecological country park consolidation model, which can effectively serve the urban ecological construction. ② There are nine functional zonings for rural development and cultivated land conservation consolidation, and their consolidation path should be based on a comprehensive rural improvement model that enhances the quality of the village and the development, utilization and protection of cultivated land resources. ③ There are 18 functional zonings for cultivated land improvement and ecological protection land consolidation, and the main consolidation path should focus on the cultivated land ecological improvement mode that emphasizes the quality of cultivated land and the improvement of regional ecological functions. ④ There are four functional zonings for ecological conservation and fallow recuperative land consolidation, and their main consolidation path should be the land ecological restoration and improvement model of construction land reclamation, cultivated land ecological conservation, and conversion of farmland to forest and grassland. The research results can provide references for Dongying City to formulate land and space planning, and can be extended to the design of comprehensive land remediation projects in other regions. It is of great significance to promote regional sustainable and scientific development.

How Do Urban Land Expansion, Land Finance, and Economic Growth Interact?

Land finance has consumed a lot of China's urban land resources while contributing to its economic growth. Urban land expansion, land finance, and economic growth have attracted significant scholarly and social attention. However, the influence mechanisms among them have not yet been fully investigated. Based on a conceptual framework analysis, in this study, the panel unit-root test, system-GMM, panel Granger causality test, impulse-response analysis, and variance decomposition were used to analyze the interactional relationships among urban land expansion, land finance, and economic growth for 30 provinces in mainland China during the period of 2000-2017. The findings show that these three factors interact with each other. Land finance exhibits a positive effect on urban land expansion and economic growth. This result is further supported by the Granger causality tests. Moreover, the VAR Granger causality-test results show a unidirectional causality flowing from urban land expansion to economic growth. The impulse-response analysis also reveals that the responses of urban land expansion to shocks in land finance appear to be positive throughout the 10 periods, which is similar to the reaction of economic growth to shocks in land finance. The result of variance decomposition indicates that the explanatory power of urban land expansion for land finance increased from 0.20% to 1.90%. In contrast, the changes in economic growth made the lowest contributions to urban land expansion and land finance. The latter made the highest contribution to economic growth, with average contribution rate of 65.26%. The findings of this study provide valuable policy implications for China, heading for a high-quality development stage.

Effect Mechanism of Land Consolidation on Soil Bacterial Community: A Case Study in Eastern China.

Farmland consolidation is an effective tool to improve farmland infrastructures, soil quality, and sustain a healthy farmland ecosystem and rural population, generating contributions to food security and regional sustainable development. Previous studies showed that farmland consolidation regulates soil physical and chemical properties. Soil microorganisms also play an important role in soil health and crop performance; however, few studies reported how farmland consolidation influence soil microecology. Here, we used DNA sequencing technology to compare bacterial community structure in farmlands with and without consolidation. DNA sequencing technology is the most advanced technology used to obtain biological information in the world, and it has been widely used in the research of soil micro-ecological environment. In September 2018, we collected soil samples in Jiashan County, Zhejiang Province, China, and used DNA sequence technology to compare the bacterial community structure in farmlands with and without consolidation. Our results found that (1) farmland consolidation had significant impacts on soil microbial characteristics, which were mainly manifested as changes in microbial biomass, microbial diversity and community structure. Farmland consolidation can increase the relative abundance of the three dominant bacteria phyla and the three fungal dominant phyla, but it also negatively affects the relative abundance of the six dominant bacteria phyla and the three fungal dominant phyla. (2) Farmland consolidation had an indirect impact on soil bacterial community structure by adjusting the soil physical and chemical properties. (3) The impact of heavy metals on bacterial community structure varied significantly under different levels of heavy metal pollution in farmland consolidation areas. There were 6, 3, 3, and 5 bacterial genera that had significant correlations with heavy metal content in cultivated land with low pollution, light pollution, medium pollution, and heavy pollution, respectively. The number of heavy metal-tolerant bacteria in the soil generally increased first and then decreased under heavy metal polluted conditions. Our study untangled the relationship between varied farmland consolidation strategies and bacteria through soil physcicochemical properties and metal pollution conditions. Our results can guide farmland consolidation strategies and sustain soil health and ecological balance in agriculture.

Farmland Transfer, Social Security, and Households' Productive Investment: Based on China's CFPS Survey.

The willingness of farmers to transfer land on a big scale will be impacted when the rural social security system is not ideal, which will limit households' productive investment. This paper investigated the intermediate effects of social security on farmland transfer and productive investment by using zero-inflated models based on 4703 samples across China. Here are the findings: (1) Farmland transfer does not significantly impact productive investment without considering social society. (2) With the improvement in social security, farmland transfer significantly affects fixed investment but is not the same as households' current investment. (3) Under the social security constraints, there is an inverted U-shaped relationship between farmland transfer and current investment. (4) The partial effect of farmland transfer on fixed investment is significantly positive, and it shows a trend of rising volatility. The government should re-examine the expected effects of the farmland transfer policy and focus on the farmers' worries about the future. Meanwhile, it is necessary to comprehensively improve the social security system and improve the multi-dimensional survival ability of farmers to give full play to the critical role of farmland transfer in current investment.

Assessing the threshold effects of road infrastructure construction on farmland use transition: an empirical study in China.

China's rapid road infrastructure construction (RIC) occupies farmland and promotes the flow of rural development factors, which causes farmland use transition (FUT). Although prior research on RIC and their impact on FUT has attracted considerable attention, few studies have investigated the threshold effect of RIC on FUT. Threshold effect is the phenomenon that when the level of RIC reaches a certain critical value, the level of FUT changes abruptly. This paper uses China's provincial panel data from 2004 to 2018 to evaluate FUT. We also consider the spatial spillover effect of RIC and construct a panel threshold regression model to assess the impact of RIC on FUT. The results demonstrate that, considering the spatial spillover effect, RIC significantly promotes FUT, and the threshold effect first rises and then falls. Also, the threshold effect of RIC on FUT also has significant regional heterogeneity. There is a significant single threshold effect for RIC in central and western China, with threshold values of 0.90 and 0.84, respectively, while the spatial lag of RIC in eastern China has a single threshold with a value of 1.10. Our research indicates that the Chinese central government should promote the synergistic increase in RIC and sustainable farmland use by improving the accessibility of road infrastructure and basing RIC planning on the actual development needs of the region.

A rare case of 17α-hydroxylase/17, 20-lyase deficiency: Clinical and genetic findings and follow-up outcomes.

Here, we reported a case of a 16-year-old Chinese female patient (46, XX) diagnosed as 17α-hydroxylase/17, 20-lyase deficiency (17-OHD) in June 2018 and over 3 years follow-up outcomes; 17-OHD is a rare form of congenital adrenal hyperplasia. The patient presented with primary amenorrhea, underdeveloped secondary sexual characteristics, hypertension and hypokalemia. Hormonal findings revealed decreased estrogen and androgen, increased progesterone, low cortisol concentration and compensatory high adrenocorticotropic hormone level. Mutation analysis of the CYP17A1 gene identified the c.1459_1467del GACTCTTTC homozygous deletion in exon 8, namely, D487_F489del mutation, resulting in the deletion of Aspartate-Serine-Phenylalanine amino acids. The patient's father and mother were all heterozygous carriers of this mutation. The diagnosis and follow-up outcomes provided useful insights to support clinical decision-making and appropriate treatment.

Improving Medical Student Clinical Knowledge and Skills Through Influenza Education.

BACKGROUND: This project aimed to implement a rigorous evaluation of influenza vaccine education as a learning tool for influenza and medical knowledge and clinical proficiency. METHODS: Among 280 Stony Brook University first-year medical students, 80 were randomly selected to participate in the Stony Brook influenza vaccine education program. Participants completed an anonymous pre-survey assessing participants': (1) experience and attitudes towards flu vaccines, (2) knowledge base of the flu virus and vaccine, (3) self-rated knowledge of the flu virus and vaccine, and (4) self-rated proficiency of clinical skills relevant to administering flu vaccines. Students then completed an educational module and vaccinated either employee healthcare workers or patients at a student-run free health clinic. Following the vaccination experience, participants completed a post-survey including questions identical to those on the pre-survey and questions regarding their evaluation of the flu vaccine education program. The pre- and post-survey data were paired, established through matching surrogate study identification codes, and differences between survey responses were analyzed using paired t-tests. RESULTS: Eighty first-year medical students participated in the pre-survey, while 55 participated in the post-survey. Compared to the pre-survey, participants significantly improved their knowledge base related to the flu virus and vaccine as well as their self-rated knowledge and clinical skills, in the post-survey. CONCLUSIONS: The Stony Brook influenza vaccination program succeeded in establishing medical student training and practice with service learning. Our study is the first to provide quantitative evidence of influenza vaccine education programs improving medical student knowledge and clinical skills. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40670-021-01355-2.

Downregulating miR-96-5p promotes proliferation, migration, and invasion, and inhibits apoptosis in human trophoblast cells via targeting DDAH1.

Several microRNAs (miRs) have been found to have modulating effects on trophoblast functions, yet the biological role and function of miR-96-5p and its interaction with Dimethylarginine Dimethylaminohydrolase 1 (DDAH1) remained poorly understood. After lentivirus transfection, the proliferation, migration, invasion and apoptosis of human trophoblast cells HTR-8/SVneo and SGHPL-4 were determined by Cell Counting Kit-8 (CCK-8) assay, scratch assay, Transwell, and flow cytometry, respectively. Relative expressions of miR-96-5p, DDAH1, and apoptosis-related proteins (B-cell lymphoma 2, Bcl-2; Bcl-2-associated X protein, Bax; cleaved (C) caspase-3) were detected via quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as needed. The target gene of miR-96-5p and their potential binding sites were predicted using TargetScan V7.2 and confirmed by dual-luciferase reporter assay. MiR-96-5p downregulation promoted proliferation, migration and invasion, suppressed apoptosis, and decreased miR-96-5p expression in trophoblast cells in vitro, while miR-96-5p upregulation had the opposite effects. DDAH1 was recognized as a target gene of miR-96-5p, and silencing DDAH1 reversed the effects of miR-96-5p downregulation on the proliferation, migration, invasion and apoptosis of trophoblast cells as well as the expressions of apoptosis-related proteins. MiR-96-5p downregulation promotes proliferation, migration, and invasion, and suppresses apoptosis in human trophoblast cells in vitro via targeting DDAH1, which provides evidence for the implication of miR-96-5p in the functional modulation of trophoblasts.

Heat shock factor 4 regulates lysosome activity by modulating the αB-crystallin-ATP6V1A-mTOR complex in ocular lens.

BACKGROUND: Germline mutations in heat shock factor 4 (HSF4) cause congenital cataracts. Previously, we have shown that HSF4 is involved in regulating lysosomal pH in mouse lens epithelial cell in vitro. However, the underlying mechanism remains unclear. METHODS: HSF4-deficient mouse lens epithelial cell lines and zebrafish were used in this study. Immunoblotting and quantitative RT-PCR were used for expression analysis. The protein-protein interactions were tested with GST-pull downs. The lysosomes were fractioned by ultracentrifugation. RESULTS: HSF4 deficiency or knock down of αB-crystallin elevates lysosomal pH and increases the ubiquitination and degradation of ATP6V1A by the proteasome. αB-crystallin localizes partially in the lysosome and interacts solely with the ATP6V1A protein of the V1 complex of V-ATPase. Furthermore, αB-crystallin can co-precipitate with mTORC1 and ATP6V1A in GST pull down assays. Inhibition of mTORC1 by rapamycin or siRNA can lead to dissociation of αB-crystallin from the ATP6V1A and mTORC1complex, shortening the half-life of ATP6V1A and increasing the lysosomal pH. Mutation of ATP6V1A/S441A (the predicted mTOR phosphorylation site) reduces its association with αB-crystallin. In the zebrafish model, HSF4 deficiency reduces αB-crystallin expression and elevates the lysosomal pH in lens tissues. CONCLUSION: HSF4 regulates lysosomal acidification by controlling the association of αB-crystallin with ATP6V1A and mTOR and regulating ATP6V1A protein stabilization. GENERAL SIGNIFICANCE: This study uncovers a novel function of αB-crystallin, demonstrating that αB-crystallin can regulate lysosomal ATP6V1A protein stabilization by complexing to ATP6V1A and mTOR. This highlights a novel mechanism by which HSF4 regulates the proteolytic process of organelles during lens development.

Knockout of DNase1l1l abrogates lens denucleation process and causes cataract in zebrafish.

Removal of nuclei in lens fiber cells is required for organelle-free zone (OFZ) formation during lens development. Defect in degradation of nuclear DNA leads to cataract formation. DNase2ß degrades nuclear DNA of lens fiber cells during lens differentiation in mouse. Hsf4 is the principal heat shock transcription factor in lens and facilitates the lens differentiation. Knockout of Hsf4 in mouse and zebrafish resulted in lens developmental defect that was characterized by retaining of nuclei in lens fiber cells. In previous in vitro studies, we found that Hsf4 promoted DNase2ß expression in human and mouse lens epithelial cells. In this study, it was found that, instead of DNase2ß, DNase1l1l is uniquely expressed in zebrafish lens and was absent in Hsf4-/- zebrafish lens. Using CRISPR-Cas9 technology, a DNase1l1l knockout zebrafish line was constructed, which developed cataract. Deletion of DNase1l1l totally abrogated lens primary and secondary fiber cell denucleation process, whereas had little effect on the clearance of other organelles. The transcriptional regulation of DNase1l1l was dramatically impaired in Hsf4-/- zebrafish lens. Rescue of DNase1l1l mRNA into Hsf4-/- zebrafish embryos alleviated its defect in lens fiber cell denucleation. Our results in vivo demonstrated that DNase1l1l is the primary DNase responsible for nuclear DNA degradation in lens fiber cells, and Hsf4 can transcriptionally activate DNase1l1l expression in zebrafish.