Integrated single-cell and spatial transcriptomics reveal microenvironment disruptions by androgen in mouse ovary.

Elevated levels of androgen are risk factors for disrupted follicular maturation in the polycystic ovary syndrome (PCOS), a reproductive disease in women. As essential cell types for follicular maturation, granulosa and thecal cells respond to androgen, but their responses are unclear at the subpopulation level. Using single-cell RNA sequencing and spatial transcriptomics, we examined the subpopulation and function alterations in an androgen-induced PCOS-like mouse model. The results demonstrated that the granulosa cell subset 5 (GC5) was active in inflammation and the thecal cell subtype 2 (TC2) had an enhanced activity in lipid metabolism. The two subsets were expanded in population size and intercellular signaling pathways, such as Ptn-Ncl and Mdk-Ncl. The results reveal that androgen induced landscape and function shifts in the two cell types under the condition of impaired follicular maturation. The study characterizes the ovarian microenvironment in responses to androgen in PCOS mice.

LncRNA PTPRG-AS1 Promotes Breast Cancer Progression by Modulating the miR-4659a-3p/QPCT Axis.

Background: Overwhelming evidence has suggested that dysregulated long noncoding RNAs (lncRNAs) play a critical modulating effect in the evolution of breast cancer (BRCA). Nevertheless, the roles of lncRNA PTPRG antisense RNA 1 (PTPRG-AS1) in BRCA and the underlying mechanisms have not been experimentally validated and functionally annotated. Methods: The expression of lncRNA PTPRG-AS1 in BRCA tissues and cell lines was evaluated by reverse transcription-quantitative PCR (RT-qPCR), and by using public databases. The proliferation of BRCA cells was detected using Cell Counting Kit-8 and colony formation assays. Wound healing assay, and Transwell migration and invasion assays were carried out to explore the migratory and invasive abilities of BRCA cells. The interaction between lncRNA PTPRG-AS1, microRNA (miR)-4659a-3p and glutaminyl-peptide cyclotransferase (QPCT) was verified using RT-qPCR, dual-luciferase reporter assay and Western blotting. Results: The results showed that LncRNA PTPRG-AS1 was markedly upregulated in BRCA tissues and cell lines. Knocking down lncRNA PTPRG-AS1 significantly inhibited the proliferation, migration and invasion of BRCA cells, while overexpression of lncRNA PTPRG-AS1 enhanced the aforementioned properties of BRCA cells. Further analyses revealed that PTPRG-AS1 may act as a molecular sponge for miR-4659a-3p, thus regulating QPCT expression, therefore, acting as an oncogene in BRCA. Conclusion: Collectively, the study demonstrates that lncRNA PTPRG-AS1 may act as a competing endogenous RNA by regulating the miR-4659a-3p/QPCT axis in BRCA progression. This lncRNA could potentially be a biomarker and therapeutic target for BRCA.

Identification of key genes associated with cervical cancer based on bioinformatics analysis.

BACKGROUND: Cervical cancer has extremely high morbidity and mortality, and its pathogenesis is still in the exploratory stage. This study aimed to screen and identify differentially expressed genes (DEGs) related to cervical cancer through bioinformatics analysis. METHODS: GSE63514 and GSE67522 were selected from the GEO database to screen DEGs. Then GO and KEGG analysis were performed on DEGs. PPI network of DEGs was constructed through STRING website, and the hub genes were found through 12 algorithms of Cytoscape software. Meanwhile, GSE30656 was selected from the GEO database to screen DEMs. Target genes of DEMs were screened through TagetScan, miRTarBase and miRDB. Next, the hub genes screened from DEGs were merged with the target genes screened from DEMs. Finally, ROC curve and nomogram analysis were performed to assess the predictive capabilities of the hub genes. The expression of these hub genes were verified through TCGA, GEPIA, qRT-PCR, and immunohistochemistry. RESULTS: Six hub genes, TOP2A, AURKA, CCNA2, IVL, KRT1, and IGFBP5, were mined through the protein-protein interaction network. The expression of these hub genes were verified through TCGA, GEPIA, qRT-PCR, and immunohistochemistry, and it was found that TOP2A, AURKA as well as CCNA2 were overexpressed and IGFBP5 was low expression in cervical cancer. CONCLUSIONS: This study showed that TOP2A, AURKA, CCNA2 and IGFBP5 screened through bioinformatics analysis were significantly differentially expressed in cervical cancer samples compared with normal samples, which might be biomarkers of cervical cancer.

Adenine's impact on mice's gut and kidney varies with the dosage administered and relates to intestinal microorganisms and enzyme activities.

This study aimed to investigate the effects of different dosages of adenine on intestinal microorganisms and enzyme activities, laying the experimental groundwork for subsequent exploration of the microbial mechanisms underlying diarrhea with kidney yang deficiency syndrome. Twenty-four mice were assigned to the following four groups: the control (NC) group, low-dosage adenine (NML) group, middle-dosage adenine (NMM) group, and high-dosage adenine (NMH) group. Mice in the NML, NMM, and NMH groups received 25 mg/(kg·d), 50 mg/(kg·d), and 100 mg/(kg·d) of adenine, respectively, 0.4 mL/each, once a day for 14 days. The NC group received 0.4 mL sterile water. Parameters including body weight, rectal temperature, intestinal microorganisms, enzyme activities, and microbial activity were measured. Results indicated that mice in the experimental group displayed signs of a poor mental state, curled up with their backs arched, and felt sleepy and lazy, with sparse fur that was easily shed, and damp bedding. Some mice showed fecal adhesion contamination in the perianal and tail areas. Dosage-dependent effects were observed, with decreased food intake, body weight, rectal temperature, and microbial activity and increased water intake and fecal water content. Enzyme activity analyses revealed significantly higher activities of protease, sucrase, amylase, and cellulase in intestinal contents and lactase, sucrase, amylase, and cellulase in the mucosa of the NMM group compared to those of other groups. Ultimately, the higher adenine dosage was associated with more pronounced symptoms of kidney yang deficiency syndrome, with 50 mg/kg adenine exhibiting the most substantial impact on the number of intestinal microbial colonies and enzyme activities.

Phosphorus Addition Reduces Seedling Growth and Survival for the Arbuscular Mycorrhizal Tree Cinnamomum camphora (Lauraceae) and Ectomycorrhizal Tree Castanopsis sclerophylla (Fagaceae) in Fragmented Forests in Eastern China.

Global changes in nutrient deposition rates and habitat fragmentation are likely to have profound effects on plant communities, particularly in the nutrient-limited systems of the tropics and subtropics. However, it remains unclear how increased phosphorus (P) supply affects seedling growth in P-deficient subtropical fragmented forests. To explore this, we applied P to 11 islands in a subtropical Chinese archipelago and examined the results in combination with a contemporary greenhouse experiment to test the influence of P addition on seedling growth and survival. We measured the growth (i.e., base area) and mortality rate of seedlings for one arbuscular mycorrhizal (AM) and one ectomycorrhizal (EcM) tree species separately and calculated their relative growth rate and mortality when compared with P addition and control treatment on each island. We also measured three functional traits and the biomass of seedlings in the greenhouse experiment. Results showed that P addition significantly increased the mortality of AM and EcM seedlings and reduced the growth rate of EcM seedlings. The relative growth rate of AM seedlings, but not EcM seedlings, significantly decreased as the island area decreased, suggesting that P addition could promote the relative growth rate of AM seedlings on larger islands. The greenhouse experiment showed that P addition could reduce the specific root length of AM and EcM seedlings and reduce the aboveground and total biomass of seedlings, indicating that P addition may affect the resource acquisition of seedlings, thereby affecting their survival and growth. Our study reveals the synergistic influence of habitat fragmentation and P deposition, which may affect the regeneration of forest communities and biodiversity maintenance in fragmented habitats.

miR136 regulates proliferation and differentiation of small tail han sheep preadipocytes.

Low meat performance is the defect of Small Tail Han sheep. Intramuscular fat affects meat quality and largely determined by adipogenesis. In previous study, miR136 was showed one of differentially expressed microRNAs between preadipocytes and mature adipocytes of Small Tail Han sheep but its role in adipogenesis is still not elucidated. Here, we investigated the effect of miR136 on adipogenesis and the underlying mechanism. qPCR data showed that miR136 level increased with preadipocytes proliferation while declined with preadipocytes differentiation. Moreover, miR136 mimics blocked lipid droplet formation, reduced lipid content and triglyceride accumulation while miR136 inhibitor showed the opposite effects, revealing that miR136 promoted preadipocytes proliferation but inhibited preadipocytes differentiation. Bioinformatics and biochemical validation manifested that PPARGC1B was a target of miR136. Furthermore, miR136 mimics decreased PPARγ and C/EBPα expression accompanied by PPARGC1B expression descending. Reverse effects were observed with miR136 inhibitor. Besides, overexpression of miR136 elevated IGF1 expression. Collectively, our data first exhibited a regulatory role of miR136 in adipogenesis, which is promoting preadipocytes proliferation through elevating IGF1 expression while inhibiting preadipocytes differentiation through targeting PPARGC1B and further declined PPARγ and C/EBPα expression. The modulation of PPARGC1B by miR136 may provide a new potential target for increasing intramuscular fat.

The dysfunction in intestinal microorganisms and enzyme activity as significant contributors to diarrhea with kidney-yang deficiency syndrome.

Object: To investigate the pathogenesis of diarrhea with kidney-yang deficiency syndrome by examining characteristic changes in intestinal microorganisms, enzyme activities, oxidative stress, and metabolism indices. Methods: Twenty mice were randomly and equally divided into control group (NC) and model group (NM). Mice in NM group received adenine suspension at a dosage of 50 mg/(kg⋅day) by gavage, 0.4 mL/time, once a day for 14 days, and Folium sennae decoction at a dosage of 10 g/(kg⋅day) by gavage, 0.4 mL/time, once a day for 7 days, starting on 8th day. Mice in NC group were administered an equivalent amount of sterile water by gavage once a day for 7 days, and twice a day from the 8th day. After modeling, assessments encompassed microbial culture, organ index calculation, microbial and enzyme activity detection, malondialdehyde (MDA) content determination, superoxide dismutase (SOD) activity, blood biochemical tests, and observation of kidney tissue pathological changes. Results: The results showed that in NM group, a reduction in the number of Lactobacillus and Bifidobacteria was noted, accompanied by an increase in the number of bacteria and E. coli. Xylanase activity in the intestinal contents and mucosa, protease activity in the intestinal mucosa, and intestinal mucosa microbial activity were diminished. Conversely, the activities of amylase, sucrase, and lactase increased in intestinal mucosa. Additionally, there was an elevation in the level of MDA. Renal tubular dilatation and inflammatory cell infiltration were observed in the renal interstitium. Conclusion: These dysfunctions in intestinal microorganisms and enzyme activities suggest potential involvement in diarrhea with kidney-yang deficiency syndrome.

miR-182-5p attenuates Schistosoma japonicum-induced hepatic fibrosis by targeting tristetraprolin.

Egg granuloma formation in the liver is the main pathological lesion caused by Schistosoma japonicum infection, which generally results in liver fibrosis and may lead to death in advanced patients. MicroRNAs (miRNAs) regulate the process of liver fibrosis, but the putative function of miRNAs in liver fibrosis induced by S. japonicum infection is largely unclear. Here, we detect a new miRNA, miR-182-5p, which shows significantly decreased expression in mouse livers after stimulation by soluble egg antigen (SEA) of S. japonicum or S. japonicum infection. Knockdown or overexpression of miR-182-5p in vitro causes the increased or decreased expression of tristetraprolin (TTP), an important immunosuppressive protein in the process of liver fibrosis. Furthermore, knockdown of miR-182-5p in vivo upregulates TTP expression and significantly alleviates S. japonicum-induced hepatic fibrosis. Our data demonstrate that downregulation of miR-182-5p increases the expression of TTP in mouse livers following schistosome infection, which leads to destabilization of inflammatory factor mRNAs and attenuates liver fibrosis. Our results uncover fine-tuning of liver inflammatory reactions related to liver fibrosis caused by S. japonicum infection and provide new insights into the regulation of schistosomiasis-induced hepatic fibrosis.

TTP protects against acute liver failure by regulating CCL2 and CCL5 through m6A RNA methylation.

Tristetraprolin (TTP), an important immunosuppressive protein regulating mRNA decay through recognition of the AU-rich elements (AREs) within the 3'-UTRs of mRNAs, participates in the pathogenesis of liver diseases. However, whether TTP regulates mRNA stability through other mechanisms remains poorly understood. Here, we report that TTP was upregulated in acute liver failure (ALF), resulting in decreased mRNA stabilities of CCL2 and CCL5 through promotion of N6-methyladenosine (m6A) mRNA methylation. Overexpression of TTP could markedly ameliorate hepatic injury in vivo. TTP regulated the mRNA stabilization of CCL2 and CCL5. Interestingly, increased m6A methylation in CCL2 and CCL5 mRNAs promoted TTP-mediated RNA destabilization. Moreover, induction of TTP upregulated expression levels of WT1 associated protein, methyltransferase like 14, and YT521-B homology N6-methyladenosine RNA binding protein 2, which encode enzymes regulating m6A methylation, resulting in a global increase of m6A methylation and amelioration of liver injury due to enhanced degradation of CCL2 and CCL5. These findings suggest a potentially novel mechanism by which TTP modulates mRNA stabilities of CCL2 and CCL5 through m6A RNA methylation, which is involved in the pathogenesis of ALF.

An NF-κB-responsive long noncoding RNA, PINT, regulates TNF-α gene transcription by scaffolding p65 and EZH2.

Long noncoding RNAs (lncRNAs) are central regulators of the inflammatory response and play an important role in inflammatory diseases. PINT has been reported to be involved in embryonic development and tumorigenesis. However, the potential functions of PINT in the innate immune system are largely unknown. Here, we revealed the transcriptional regulation of inflammatory genes by PINT, whose expression is primarily dependent on the NF-κB signaling pathway in human and mouse macrophage and intestinal epithelial cell lines. Functionally, PINT selectively regulates the expression of TNF-α in basal and LPS-stimulated cells. Mechanistically, PINT acts as a modular scaffold of p65 and EZH2 to coordinate their localization and specify their binding to the target genes. Further, a high expression level of PINT was detected in intestinal mucosal tissues from patients with ulcerative colitis (UC). Together, these findings demonstrate that PINT acts as an activator of inflammatory responses, highlighting the importance of this lncRNA as a potential therapeutic target in infectious diseases and inflammatory diseases.

Zn-Cu-In-S-Se Quinary "Green" Alloyed Quantum-Dot-Sensitized Solar Cells with a Certified Efficiency of 14.4 .

The photoelectronic properties of quantum dots (QDs) have a critical impact on the performance of quantum-dot-sensitized solar cells (QDSCs). Currently, I-III-VI group QDs have become the mainstream light-harvesting materials in high-performance QDSCs. However, it is still a great challenge to achieve satisfactory efficiency for light-harvesting, charge extraction, and charge collection simultaneously in QDSCs. We design and prepare Zn0.4 Cu0.7 In1.0 Sx Se2-x (ZCISSe) quinary alloyed QDs by cation/anion co-alloying strategy. The critical photoelectronic properties of target QDs, including band gap, conduction band energy level, and density of defect trap states, can be conveniently tailored. Experimental results demonstrate that the ZCISSe quinary alloyed QDs can achieve an ideal balance among light-harvesting, photogenerated electron extraction, and charge-collection efficiencies in QDSCs compared to its single anion or cation quaternary alloyed QD counterparts. Consequently, the quinary alloyed QDs boost the certified efficiency of QDSCs to 14.4 %, which is a new efficiency record for liquid-junction QD solar cells.

Upregulation of KSRP by miR-27b attenuates schistosomiasis-induced hepatic fibrosis by targeting TGF-ß1.

Hepatic stellate cells (HSCs) are the main effectors for various types of hepatic fibrosis, including Schistosome-induced hepatic fibrosis. Multiple inflammatory cytokines/chemokines, such as transforming growth factor-ß1 (TGF-ß1), activate HSCs, and contribute to the development of hepatic fibrosis. MicroRNAs regulate gene expression at the posttranscriptional level and are involved in regulation of inflammatory cytokine/chemokine synthesis. In this study, we showed that soluble egg antigen (SEA) stimulation and Schistosoma japonicum infection downregulate miR-27b expression and increase KH-type splicing regulatory protein (KSRP) mRNA and protein levels in vitro and in vivo. miR-27b regulates the stabilization of TGF-ß1 mRNA through targeting KSRP by interacting with their AU-rich elements in hepatocytes and non-parenchymal cells, which has an effect on the activation of HSCs. Importantly, our results have shown that either knockdown miR-27b or overexpression of KSRP attenuates S. japonicum-induced hepatic fibrosis in vivo. Therefore, our study highlights the crucial role of miR-27b and KSRP in the negative regulation of immune reactions in hepatocyte and non-parenchymal cells in response to SEA stimulation and S. japonicum infection. It reveals that manipulation of miR-27b or KSRP might be a useful strategy not only for treating Schistosome-induced hepatic fibrosis but also for curing hepatic fibrosis in general.

[Structure and function of B-cell linker and its role in the development of B cell-related diseases].

B cell linker (BLNK) is a key linker protein of B cell receptor (BCR) signaling pathway. BLNK participates in the regulation of PLC-γactivity and the activation of Ras pathway through its typical structure and interaction network with other proteins, and is thus widely involved in the regulation of B cell proliferation, differentiation, apoptosis and signal transduction. Furthermore, it is closely related to anaphylactic diseases, multiple sclerosis, chromosomal aneuploidy, aneuglobulinemia, B lymphocytic leukemia and lymphoma. Herein we review the structure and biological function of BLNK and its role in B cell-related diseases. BLNK can cooperate with a series of effective proteins to activate BCR signaling pathway, thereby regulating the development, maturation and function of B cells. The functional mutation of BLNK can destroy the homeostasis of B cells and affect the development and maturation of B cells, which leads to the occurrence of B cell related diseases. A comprehensive understanding of the biological functions of BLNK not only provides insights into the pathogenesis of B cell-related diseases, but also inspires new ideas and helps to find breakthroughs for the treatment of these diseases with BLNK as the therapeutic target.

Enhancing Loading Amount and Performance of Quantum-Dot-Sensitized Solar Cells Based on Direct Adsorption of Quantum Dots from Bicomponent Solvents.

Intrinsically weak interaction between oil-soluble quantum dots (QDs) and TiO2 in a direct adsorption process limits QD loading and the performance of QD-sensitized solar cells (QDSCs). Herein, the underlying chemistry and mechanisms governing QD adsorption on TiO2 were studied to improve QD loading and cell performance. Experimental results indicate that solvent polarity plays the crucial role in determining QD loading. Compared with single-component solvents, substantially greater QD loading can be realized at the critical point (CP) of bicomponent solvents, where QDs become metastable and start to precipitate. Through this strategy, average efficiency of 12.24% was obtained for ZCISe QDSCs, which is comparable to those based on the capping ligand induced self-assembly route. This report demonstrates the great potential of bicomponent solvents at the CP for high QD loading and excellent cell performance and presents a platform for assembling functional composites with the use of different nanocrystals and substrates.

Origin of the effects of PEG additives in electrolytes on the performance of quantum dot sensitized solar cells.

It has been well established that polymer additives in electrolyte can impede the charge recombination processes at the photoanode/electrolyte interface, and improve performance, especially V oc, of the resulting sensitized solar cells. However, there are few reports about the effect of electrolyte additives on counter electrode (CE) performance. Herein, we systematically investigated the effect of polyethylene glycol (PEG) additives with various molecular weights (M w from 300 to 20 000) in polysulfide electrolyte on the performance of two representative CdSe and Zn-Cu-In-Se (ZCISe) quantum dot sensitized solar cells (QDSCs), and explored the mechanism of the observed effects. Electrochemical impedance spectroscopy measurements indicate that all PEG additives can improve the charge recombination resistance at the photoanode/electrolyte interface, therefore suppressing the unwanted charge recombination process, and enhancing the V oc of the resulting cell devices accordingly. On the CE side, with the increase of M w of PEG additives, the initial effect of reducing the charge transfer resistance at the CE/electrolyte interface evolves into an increasing resistance; accordingly the initial positive effect on FF turns into negative one. Accordingly, low M w PEG can improve efficiency for both CdSe (increasing from 6.81% to 7.60%) and ZCISe QDSCs (increasing from 9.26% to 10.20%). High M w PEG is still effective for CdSe QDSCs with an efficiency of 7.38%, but falls flat on ZCISe QDSCs (with an efficiency of 9.11%).

Comparative study on the treatment of raw and biologically treated textile effluents through submerged nanofiltration.

Raw and biologically treated textile effluents were submerged filtrated using lab-fabricated hollow fiber nanofiltration membrane with a molecular weight cut-off of about 650 g/mol. Permeate flux, chemical oxygen demand (COD) reduction, color removal, membrane fouling, and cleaning were investigated and compared by varying the trans-membrane pressure (TMP) and volume concentrating factor (VCF). It was found that both raw and biologically treated textile effluents could be efficiently treated through submerged nanofiltration. The increase of TMP resulted in a decline in water permeability, COD reduction, color removal, and flux recovery ratio, while the increase of VCF resulted in both increased COD reduction and color removal. Under the TMP of 0.4 bar and VCF of 5.0, fluxes of 1.96 and 2.59 l/m(2)h, COD reductions of 95.7 and 94.2%, color removals of 99.0, and 97.3% and flux recovery ratios of 91.1 and 92.9% could be obtained in filtration of raw and biologically treated effluents, respectively. After filtration, the COD and color contents of the raw effluent declined sharply from 1780 to 325 mg/l and 1.200 to 0.060 Abs/cm, respectively, while for the biologically treated effluent, they decreased from 780 to 180 mg/l and 0.370 to 0.045 Abs/cm, respectively.