Adsorption and desorption characteristics of flavonoids from white tea using macroporous adsorption resin.

White tea contains the highest flavonoids compared to other teas. While there have been numerous studies on the components of different tea varieties, research explicitly focusing on the flavonoid content of white tea remains scarce, making the need for a good flavonoid purification process for white tea even more important. This study compared the adsorption and desorption performance of five types of macroporous resins: D101, HP20, HPD500, DM301, and AB-8. Among the tested resins, AB-8 was selected based on its best adsorption and desorption performance to investigate the static adsorption kinetics and dynamic adsorption-desorption purification of white tea flavonoids. The optimal purification process was determined: adsorption temperature 25 °C, crude tea flavonoid extract pH 3, ethanol concentration 80 %, sample loading flow rate and eluent flow rate 1.5 BV/min, and eluent dosage 40 BV. The results indicated that the adsorption process followed pseudo-second-order kinetics. Under the above purification conditions, the purity of the total flavonoids in the purified white tea flavonoid increased from approximately 17.69 to 46.23 %, achieving a 2.61-fold improvement, indicating good purification results. The purified white tea flavonoid can be further used for nutraceutical and pharmaceutical applications.

PUF60 promotes cell cycle and lung cancer progression by regulating alternative splicing of CDC25C.

Alternative splicing (AS) has been implicated in cell cycle regulation and cancer, but the underlying mechanisms are poorly understood. The poly(U)-binding splicing factor 60 (PUF60) is essential for embryonic development and is overexpressed in multiple types of cancer. Here, we report that PUF60 promotes mitotic cell cycle and lung cancer progression by controlling AS of the cell division cycle 25C (CDC25C). Systematic analysis of splicing factors deregulated in lung adenocarcinoma (LUAD) identifies that elevated copy number and expression of PUF60 correlate with poor prognosis. PUF60 depletion inhibits LUAD cell-cycle G2/M transition, cell proliferation, and tumor development. Mechanistically, PUF60 knockdown leads to exon skipping enriched in mitotic cell cycle genes, including CDC25C. Exon 3 skipping in the full-length CDC25C results in nonsense-mediated mRNA decay and a decrease of CDC25C protein, thereby inhibiting cell proliferation. This study establishes PUF60 as a cell cycle regulator and an oncogenic splicing factor in lung cancer.

Ethanol Causes Cell Death and Neuronal Differentiation Defect During Initial Neurogenesis of the Neural Retina by Disrupting Calcium Signaling in Human Retinal Organoids.

Fetal Alcohol Syndrome (FAS) affects a significant proportion, exceeding 90%, of afflicted children, leading to severe ocular aberrations such as microphthalmia and optic nerve hypoplasia. During the early stages of pregnancy, the commencement of neural retina neurogenesis represents a critical period for human eye development, concurrently exposing the developing retinal structures to the highest risk of prenatal ethanol exposure due to a lack of awareness. Despite the paramount importance of this period, the precise influence and underlying mechanisms of short-term ethanol exposure on the developmental process of the human neural retina have remained largely elusive. In this study, we utilize the human embryonic stem cells derived retinal organoids (hROs) to recapitulate the initial retinal neurogenesis and find that 1% (v/v) ethanol slows the growth of hROs by inducing robust cell death and retinal ganglion cell differentiation defect. Bulk RNA-seq analysis and two-photon microscope live calcium imaging reveal altered calcium signaling dynamics derived from ethanol-induced down-regulation of RYR1 and CACNA1S. Moreover, the calcium-binding protein RET, one of the downstream effector genes of the calcium signaling pathway, synergistically integrates ethanol and calcium signals to abort neuron differentiation and cause cell death. To sum up, our study illustrates the effect and molecular mechanism of ethanol on the initial neurogenesis of the human embryonic neural retina, providing a novel interpretation of the ocular phenotype of FAS and potentially informing preventative measures for susceptible populations.

Ammonia mitigation campaign with smallholder farmers improves air quality while ensuring high cereal production.

Reducing cropland ammonia (NH3) emissions while improving air quality and food supply is a challenge, particularly in China where there are millions of smallholder farmers. We tested the effectiveness of a tailored nitrogen (N) management strategy applied to wheat-maize cropping systems in 'demonstration squares' across Quzhou County in the North China Plain. The N-management techniques included optimal N rates, deep fertilizer placement and application of urease inhibitors, implemented through cooperation between government, researchers, businesses and smallholders. Compared with conventional local smallholder practice, our NH3 mitigation campaign reduced NH3 volatilization from wheat and maize by 49% and 39%, and increased N-use efficiency by 28% and 40% and farmers' profitability by 25% and 19%, respectively, with no detriment to crop yields. County-wide atmospheric NH3 and fine particulate matter (with aerodynamic diameter <2.5 µm) concentrations decreased by 40% and 8%, respectively. County-wide net benefits were estimated at US$7.0 million. Our demonstration-square approach shows that cropland NH3 mitigation and improved air quality and farm profitability can be achieved simultaneously by coordinated actions at the county level.

A controllable perfusion microfluidic chip for facilitating the development of retinal ganglion cells in human retinal organoids.

Retinal organoids (ROs) derived from human pluripotent stem cells (hPSCs) have become a promising model in vitro to recapitulate human retinal development, which can be further employed to explore the mechanisms of retinal diseases. However, the current culture systems for ROs lack physiologically relevant microenvironments, such as controllable mechano-physiological cues and dynamic feedback between cells and the extracellular matrix (ECM), which limits the accurate control of RO development. Therefore, we designed a controllable perfusion microfluidic chip (CPMC) with the advantages of precisely controlling fluidic shear stress (FSS) and oxygen concentration distribution in a human embryonic stem cell (hESC)-derived RO culture system. We found that ROs cultured under this system allow for expanding the retinal progenitor cell (RPC) pool, orchestrating the retinal ganglion cell (RGC) specification, and axon growth without disturbing the spatial and temporal patterning events at the early stage of RO development. Furthermore, RNA sequencing data revealed that the activation of voltage-gated ion channels and the increased expression of ECM components synergistically improve the growth of ROs and facilitate the differentiation of RGCs. This study elaborates on the advantages of the designed CPMC to promote RO growth and provide a controllable and reliable platform for the efficient maturity of RGCs in the ROs, promising applications in modeling RGC-related disorders, drug screening, and cell transplantation.

Application of Human Stem Cell Derived Retinal Organoids in the Exploration of the Mechanisms of Early Retinal Development.

The intricate neural circuit of retina extracts salient features of the natural world and forms bioelectric impulse as the origin of vision. The early development of retina is a highly complex and coordinated process in morphogenesis and neurogenesis. Increasing evidence indicates that stem cells derived human retinal organoids (hROs) in vitro faithfully recapitulates the embryonic developmental process of human retina no matter in the transcriptome, cellular biology and histomorphology. The emergence of hROs greatly deepens on the understanding of early development of human retina. Here, we reviewed the events of early retinal development both in animal embryos and hROs studies, which mainly comprises the formation of optic vesicle and optic cup shape, differentiation of retinal ganglion cells (RGCs), photoreceptor cells (PRs) and its supportive retinal pigment epithelium cells (RPE). We also discussed the classic and frontier molecular pathways up to date to decipher the underlying mechanisms of early development of human retina and hROs. Finally, we summarized the application prospect, challenges and cutting-edge techniques of hROs for uncovering the principles and mechanisms of retinal development and related developmental disorder. hROs is a priori selection for studying human retinal development and function and may be a fundamental tool for unlocking the unknown insight into retinal development and disease.

[Emission Inventory and Distribution Characteristics of Anthropogenic Ammonia in Quzhou County, North China Plain].

Quzhou county is a county in the North China Plain with intensive agricultural production. Based on data collected on the local anthropogenic activity level and the ammonia (NH3) emission factor, the anthropogenic NH3 emission inventory in Quzhou county from 2002 to 2019 was established using the emission factor method, and the NH3 emission inventory in 2019 was optimized using the locally measured nitrogen (N) fertilizer application NH3 emission factor and activity data from farmer surveys. We found that the total NH3 emission in Quzhou county showed a bimodal pattern: it increased from 6682.9 t in 2002 to 7195.0 t in 2004 and then decreased to 5872.0 t in 2008. The second peak occurred in 2015 (7010.5 t) and then gradually decreased to 5636.3 t in 2018. Livestock husbandry (61%-75%) and N fertilizer application (14%-28%) were the primary NH3 emission sources. In 2019, the total NH3 emission in Quzhou county was 6559.7 t, of which N fertilizer application and livestock husbandry contributed 28% and 61%, respectively. Wheat planting contributed the highest NH3 emissions, accounting for 40% of the total NH3 emission of crop production. Laying hens had the largest NH3 emission in livestock production, contributing 40% of the total emissions. NH3 emissions were higher in the south than those in the north at the county level, with the average NH3 emission intensity reaching 13.5 t·km-2. Nanliyue and Baizhai towns were hot spots of NH3 emission. Taken together NH3 emission reduction in wheat production and layer hen breeding will help to control atmospheric NH3 pollution in the North China Plain.

Development of human retinal organoid models for bisphenol toxicity assessment.

Bisphenols, including Bisphenol A (BPA), Tetrabromobisphenol A (TBBPA), and Tetrabromobisphenol S (TBBPS), have been widely applied in the production of polycarbonate plastics and epoxy resins and have been detected in the environment worldwide. The frequent detection of bisphenols in maternal and fetal samples has raised concerns about their toxic effects on human embryonic development, especially on the development of the central nervous system. However, the effect of bisphenols on human retinal development is still unknown. In this study, to evaluate the toxicity of bisphenols on early retinal development, human embryonic stem cells were induced to differentiate into retinal organoids that responded to BPA, TBBPA, and TBBPS, at human exposure relevant concentrations. The global gene expression of retinal organoids was analyzed by RNA sequencing (RNA-seq). A set of retinal development-related biological processes, including neuron differentiation, phototransduction, axon guidance, and retina layer formation, were identified in retinal organoids corresponding to different developmental stages. The RNA-seq data also showed that BPA, TBBPA, and TBBPS influenced retinal development by interfering with the Cytokine-cytokine receptor interaction pathway. HSPA6, HIF1A-AS3, CDC20B, IL19, OAS1, HSPA7, and RN7SK were dysregulated by these chemicals. Additionally, BPA, TBBPA, and TBBPS exhibited different toxic effects on neural retina development, with TBBPA appearing to exert more toxicity than BPA and TBBPS. Furthermore, three bisphenols exhibited different effects at different stages of neural retina development. The sensitivity of retinal development to bisphenols depends on their developmental stage. This study provides new insights into the deep dissection of retinotoxicity after prenatal bisphenol exposure.

Reactive N emissions from cropland and their mitigation in the North China Plain.

Excessive application of chemical nitrogen (N) fertilizer and inefficient N management are still common in the North China Plain, leading to large reactive N (Nr) losses and pollution, threatening environmental security and public health. Three improved N management practices (33% reduction in N applied (OU), OU combined with partial organic fertilizer substitution (UOM) and the urea in UOM amended with a urease inhibitor (ULOM)) together with no N application (CK) and farmers' conventional practice (CU) were tested on a maize-wheat rotation at Quzhou, Hebei, North China Plain (NCP). Nr emissions were related to WFPS (Water Filled Pore Space), soil mineral N (NH4+-N and NO3--N) and soil temperature. Nr emissions and yield-scaled Nr emissions were significantly reduced by partial substitution of organic fertilizer for chemical fertilizer: NH3 emissions were reduced by 55.8-62.4%. Using a urease inhibitor (Limus®), further reduced NH3 emissions by 40.2-64.5%. Yield-scaled NH3 emissions were, on average, reduced by 60.0% and 55.2% in the maize and wheat growing season, respectively, relative to the UOM treatment. Long-term application of organic fertilizer had a significant positive effect on N use efficiency (NUE). Overall, the study shows that appropriated N management such as reducing the N application rate, partial substitution of chemical N by organic N and using a urease inhibitor can reduce Nr emissions and promote NUE in the North China Plain. The methods corresponding to the ULOM and UOM treatments were the most and second most effective, respectively, with high net economic benefits.

3D Printing of Multiscale Ti64-Based Lattice Electrocatalysts for Robust Oxygen Evolution Reaction.

Electrically assisted water splitting is an endurable strategy for hydrogen production, but the sluggish kinetics of oxygen evolution reaction (OER) extremely restrict the large-scale production of hydrogen. Developing highly efficient and non-precious catalytic materials is essential to accelerate the sluggish kinetics of OER. However, currently used catalyst supports, such as copper foam, suffer from inferior corrosion resistance and structural stability, resulting in the disabled functionality of 3D conductive networks. To this end, a novel 3D freestanding electrode with corrosion-resistant and robust Ti-6Al-4V titanium alloy lattice as the catalyst support is designed via a 3D printing technology of selective laser melting. After the coating of core-shell Cu(OH)2@CoNi carbonate hydroxides (CoNiCH) on the designed lattice, a unique micro/nano-sized hierarchical porous structure is formed, which endows the electrocatalyst with a promising electrocatalytic activity (a low overpotential of 355 mV at 30 mA cm-2 and Tafel slope of 125.3 mV dec-1 ). Computational results indicate that the CoNiCH exhibits optimized electron transfer and the catalytic activity of the Ni site is higher than that of the Co site in the CoNiCH. Therefore, the integration of robust catalyst supports and highly active materials opens up an avenue for reliable and high-performance OER electrocatalysts.

Single-cell RNA-sequencing of zebrafish hair cells reveals novel genes potentially involved in hearing loss.

Hair cells play key roles in hearing and balance, and hair cell loss would result in hearing loss or vestibular dysfunction. Cellular and molecular research in hair cell biology provides us a better understanding of hearing and deafness. Zebrafish, owing to their hair cell-enriched organs, have been widely applied in hair cell-related research worldwide. Similar to mammals, zebrafish have inner ear hair cells. In addition, they also have lateral line neuromast hair cells. These different types of hair cells vary in morphology and function. However, systematic analysis of their molecular characteristics remains lacking. In this study, we analyzed the GFP+ cells isolated from Tg(Brn3c:mGFP) larvae with GFP expression in all hair cells using single-cell RNA-sequencing (scRNA-seq). Three subtypes of hair cells, namely macula hair cell (MHC), crista hair cell (CHC), and neuromast hair cell (NHC), were characterized and validated by whole-mount in situ hybridization analysis of marker genes. The hair cell scRNA-seq data revealed hair cell-specific genes, including hearing loss genes that have been identified in humans and novel genes potentially involved in hair cell formation and function. Two novel genes were discovered to specifically function in NHCs and MHCs, corresponding to their specific expression in NHCs and MHCs. This study allows us to understand the specific genes in hair cell subpopulations of zebrafish, which will shed light on the genetics of both human vestibular and cochlear hair cell function.

Single Cell Transcriptome Sequencing of Zebrafish Testis Revealed Novel Spermatogenesis Marker Genes and Stronger Leydig-Germ Cell Paracrine Interactions.

Spermatogenesis in testis is an important process for sexual reproduction, and worldwide about 10-15 percent of couples suffer from infertility. It is of importance to study spermatogenesis at single cell level in both of human and model organisms. Currently, single-cell RNA sequencing technologies (scRNA-seq) had been extensively applied to the study of cellular components and its gene regulations in the testes of different species, including human, monkey, mouse, and fly, but not in zebrafish. Zebrafish was a widely used model organism in biology and had been extensively used for the study of spermatogenesis in the previous studies. Therefore, it is also important to profile the transcriptome of zebrafish testis at single cell level. In this study, the transcriptomes of 14, 315 single cells from adult male zebrafish testes were profiled by scRNA-seq, and 10 cell populations were revealed, including Leydig cell, Sertoli cell, spermatogonia cell (SPG), spermatocyte, and spermatids. Notably, thousands of cell-type specific novel marker genes were identified, including sumo3b for SPG, krt18a.1 for Sertoli cells, larp1b and edrf1 for spermatids, which were also validated by RNA in situ hybridization experiments. Interestingly, through Ligand-Receptor (LR) analyses, zebrafish Leydig cells demonstrated stronger paracrine influence on germ cells than Sertoli cells. Overall, this study could be an important resource for the study of spermatogenesis in zebrafish and might also facilitate the study of the genes associated with human infertility through using zebrafish as a model organism.

Advanced human developmental toxicity and teratogenicity assessment using human organoid models.

Tremendous progress has been made in the field of toxicology leading to the advance of developmental toxicity assessment. Conventional animal models and in vitro two-dimensional models cannot accurately describe toxic effects and predict actual in vivo responses due to obvious inter-species differences between humans and animals, as well as the lack of a physiologically relevant tissue microenvironment. Human embryonic stem cell (hESC)- and induced pluripotent stem cell (iPSC)-derived three-dimensional organoids are ideal complex and multicellular organotypic models, which are indispensable in recapitulating morphogenesis, cellular interactions, and molecular processes of early human organ development. Recently, human organoids have been used for drug discovery, chemical toxicity and safety in vitro assessment. This review discusses the recent advances in the use of human organoid models, (i.e., brain, retinal, cardiac, liver, kidney, lung, and intestinal organoid models) for developmental toxicity and teratogenicity assessment of distinct tissues/organs following exposure to pharmaceutical compounds, heavy metals, persistent organic pollutants, nanomaterials, and ambient air pollutants. Combining next-generation organoid models with innovative engineering technologies generates novel and powerful tools for developmental toxicity and teratogenicity assessment, and the rapid progress in this field is expected to continue.

Structural and functional alterations in untreated patients with major depressive disorder and bipolar disorder experiencing first depressive episode: A magnetic resonance imaging study combined with follow-up.

BACKGROUND: Magnetic resonance imaging (MRI) could assist in identifying objective biomarkers and follow-up study could effectively improve subjective diagnostic accuracy. By combining MRI with follow-up, this study aims to determine the shared and distinct alterations between major depressive disorder (MDD) and bipolar disorder (BD). METHODS: Untreated patients with MDD experiencing the first episode were subjected to MRI and subsequent follow-up. Fifteen patients with mania or hypomania were regrouped into BD group. Twenty patients were still grouped as MDD after an average of 37.95 months follow-up. Thirty healthy controls (HCs) were recruited to match the patients. Gray matter volume (GMV) and amygdala-seed functional connectivity (FC) in the whole brain were detected and compared among the three groups. RESULTS: GMV analysis revealed that the MDD and BD groups presented reduced GMV predominantly in the parietal, occipital, and frontal regions in the bilateral cerebrum compared with the HCs. The BD group had reduced GMV predominantly in the parietal, temporal, insular regions and the Rolandic operculum in the right-side cerebrum compared with MDD and HC groups. FC analysis revealed that the MDD and BD patients displayed increased FC values mainly in the bilateral parietal, and left occipital regions. Only the BD group displayed increased FC values in the temporal, occipital, parietal and limbic regions in the right-side cerebrum relative to HCs. LIMITATIONS: The main limitation is the relatively small sample size. CONCLUSIONS: Alterations in the cortical regions and cortico-limbic neural system may provide the scientific basis for differential diagnosis in affective disorders.

Gender differences of amplitude of low-frequency fluctuations in bipolar disorder: A resting state fMRI study.

BACKGROUND: The clinical and epidemiological features of bipolar disorder (BD) between females and males have many differences. The association between brain function and gender in BD is unknown. This research aimed to investigate the association between brain function and gender in BD by using amplitude of low-frequency fluctuations (ALFFs). METHODS: Ninety-eight patients (49 females and 49 males) with BD and 171 matched healthy controls (HCs, 89 females and 82 males) were recruited for resting-state functional magnetic resonance imaging. ALFF was used to estimate brain function. RESULTS: A main effect of diagnosis in ALFF was observed in the dorsal lateral prefrontal cortex (DLPFC), ventral prefrontal cortex (VPFC), caudate and occipital lobe. A main effect of gender in ALFF was found in the right VPFC, DLPFC, thalamus, and occipital lobe. A main effect of diagnosis gender interaction in ALFF was observed in the left DLPFC. Analyses of two-sample t-test indicated that male patients with BD had increased ALFF in the right hippocampus, right amygdala, left caudate, and left DLPFC, and decreased ALFF in the occipital lobe compared with male HC. Female patients with BD demonstrated increased ALFF in the right VPFC and right DLPFC compared with female HC. Male patients with BD exhibited increased ALFF in the right VPFC and left DLPFC and decreased ALFF in the occipital lobe compared with female patients with BD. LIMITATIONS: This study did not consider the effect of medications and emotional states on brain activity. CONCLUSIONS: Results suggested gender differences in the dysfunctions of the cortico-limbic neural system in BD.

Common and distinct neural activities in frontoparietal network in first-episode bipolar disorder and major depressive disorder: Preliminary findings from a follow-up resting state fMRI study.

BACKGROUND: It is difficult to distinguish bipolar disorder (BD) from major depressive disorder (MDD), especially with the initial depressive episode. In this study, we compared neural activities of BD and MDD patients during the first-episode (FE) to investigate common and distinct neural activities and further explore predictive indicators in the two diseases. METHODS: FE-MDD patients were performed resting state functional magnetic resonance imaging and followed up after scanning. After follow-up, FE-MDD patients were regrouped into FE-BD and FE-MDD patients. The study included 24 FE-BD patients, 28 FE-MDD patients, and 30 age- and sex-matched healthy controls (HC) to investigate neural activities with regional homogeneity (ReHo) analysis among the 3 groups. RESULTS: Compared to HC, FE-BD patients displayed significantly higher ReHo values in the superior frontal gyrus, the medial superior frontal gyrus within right-side cerebral hemisphere than FE-MDD patients and HC. Compared to HC, FE-BD and FE-MDD patients displayed significant decreased ReHo values in the paracentral lobule, the precuneus and the median cingulate and paracingulate gyrus within bilateral cerebral hemisphere, and the postcentral gyrus and the precentral gyrus within the right-side. FE-BD displayed significant lower ReHo values than FE-MDD patients in these regions. LIMITATIONS: The potential effects of medicine, age, course of disease and handedness on results could not be ignored. CONCLUSIONS: Abnormal neural activities of frontoparietal network may provide common and distinct markers to affective disorders and scientific basis for further prediction researches of affective disorders.

Amplitude of low-frequency fluctuation (ALFF) may be associated with cognitive impairment in schizophrenia: a correlation study.

BACKGROUND: Cognitive impairments are prominent in schizophrenia (SZ). Imaging studies have demonstrated that functional changes of several areas of the brain exist in SZ patients. The relationships between these two indexes are largely unexplored in SZ. The MATRICS Consensus Cognitive Battery (MCCB) was used to measure cognitive impairment in multi-dimensional cognitive fields of SZ patients. This study was conducted to explore the relationship between cognitive functional impairment and the amplitude of low-frequency fluctuation (ALFF) in SZ patients. METHOD: A total of 104 participants (44 SZ patients and 60 age- and gender-matched healthy controls (HC)) were recruited for this study. The MCCB was used to assess cognitive function of the participants, while brain activity was assessed using the ALFF. The relationship between the MCCB and the ALFF was investigated by using a correlation analysis. RESULTS: There were significant differences between SZ patients and HC in MCCB total and domain scores as well as in ALFF results. The reduction of ALFF in the bilateral postcentral gyri and paracentral lobule in SZ patients has a negative correlation with the MCCB sub-test of symbol coding. CONCLUSION: These findings suggest that the reduction of ALFF in bilateral postcentral gyri and paracentral lobule may be related to cognitive impairment in SZ patients.