SIRT1 inhibits apoptosis of human lens epithelial cells through suppressing endoplasmic reticulum stress in vitro and in vivo

AIM:To explore the effect of silent information regulator factor 2-related enzyme 1(SIRT1)on modulating apoptosis of human lens epithelial cells(HLECs)and alleviating lens opacification of rats through suppressing endoplasmic reticulum(ER)stress.METHODS:HLECs(SRA01/04)were treated with varying concentrations of tunicamycin(TM)for 24h,and the expression of SIRT1 and C/EBP homologous protein(CHOP)was assessed using real-time quantitative polymerase chain reaction(RT-PCR),Western blotting,and immunofluorescence.Cell morphology and proliferation was evaluated using an inverted microscope and cell counting kit-8(CCK-8)assay,respectively.In the SRA01/04 cell apoptosis model,which underwent siRNA transfection for SIRT1 knockdown and SRT1720 treatment for its activation,the expression levels of SIRT1,CHOP,glucose regulated protein 78(GRP78),and activating transcription factor 4(ATF4)were examined.The potential reversal of SIRT1 knockdown effects by 4-phenyl butyric acid(4-PBA;an ER stress inhibitor)was investigated.In vivo,age-related cataract(ARC)rat models were induced by sodium selenite injection,and the protective role of SIRT1,activated by SRT1720 intraperitoneal injections,was evaluated through morphology observation,hematoxylin and eosin(H&E)staining,Western blotting,and RT-PCR.RESULTS:SIRT1 expression was downregulated in TMinduced SRA01/04 cells.Besides,in SRA01/04 cells,both cell apoptosis and CHOP expression increased with the rising doses of TM.ER stress was stimulated by TM,as evidenced by the increased GRP78 and ATF4 in the SRA01/04 cell apoptosis model.Inhibition of SIRT1 by siRNA knockdown increased ER stress activation,whereas SRT1720 treatment had opposite results.4-PBA partly reverse the adverse effect of SIRT1 knockdown on apoptosis.In vivo,SRT1720 attenuated the lens opacification and weakened the ER stress activation in ARC rat models.CONCLUSION:SIRT1 plays a protective role against TM-induced apoptosis in HLECs and slows the progression of cataract in rats by inhibiting ER stress.These findings suggest a novel strategy for cataract treatment focused on targeting ER stress,highlighting the therapeutic potential of SIRT1 modulation in ARC development.

Tcf7l1 promotes transcription of Kruppel-like factor 4 during Xenopus embryogenesis

Kruppel-like factor 4（Klf4） is a zinc finger transcription factor and plays crucial roles in Xenopus embryogenesis.However, its regulation during embryogenesis is still unclear. Here, we report that Tcf711, a key downstream transducer of the Wnt signaling pathway, could promote Klf4 transcription and stimulate Klf4 promoter activity in early Xenopus embryos. Furthermore, cycloheximide treatment showed a direct effect on Klf4 transcription facilitated by Tcf711. Moreover, the dominant negative form of Tcf711（dnTcf711）, which lacks N-terminus of the β-catenin binding motif, could still activate Klf4 transcription, suggesting that this regulation is Wnt/β-catenin independent.Taken together, our results demonstrate that Tcf711 lies upstream of Klf4 to maintain its expression level during Xenopus embryogenesis.

Stochastic Differential Game Model for Decentralized Power Control with Wisdom Regulation Factor in Wearable Audio-Oriented BodyNets

In this study, aiming at the characteristics of randomness and dynamics in Wearable Audiooriented BodyNets （WA-BodyNets）, stochastic differential game theory is applied to the investigation of the problem of transmitted power control inconsumer electronic devices. First, astochastic differential game model is proposed for non-cooperative decentralized uplink power control with a wisdom regulation factor over WA-BodyNets with a onehop star topology.This model aims to minimize the cost associated with the novel payoff function of a player, for which two cost functions are defined： functions of inherent power radiation and accumulated power radiation darmge. Second, the feedback Nash equilibrium solution of the proposed model and the constraint of the Quality of Service （QoS） requirement of the player based on the SIR threshold are derived by solving the Fleming-Bellman-Isaacs partial differential equations. Furthermore, the Markov property of the optimal feedback strategies in this model is verified.The simulation results show that the proposed game model is effective and feasible for controlling the transmitted power of WA-BodyNets.

The Impact Heme Oxygenase -1 on the Regulating Factors of Hepatoma Cells＇ Cell Cycle

It aims to analyze the impact heme oxygenase -1 （heme oxygenase 1, HO-1） on regulating factors of human hepatoma cell HepG2＇s cell cycle, through constructing recombinant vector of pcDNA3.1 containing wild-type and mutant HO-1 gene （＋）-wtHO-1 and pcDNA3.1 （＋）-mHO-1G143H. By using the method of liposome-mediated, the recombinant vector was transfected hepatoma cell line HepG2. And the transfected one with empty vector was treated as a control group. By the selection of G418, stable expression of wild-type and mutant HO-1 in HepG2 liver cancer cell lines were established. Use the blot of semi-quantitative RT-PCR and Western to test transfected cell lines expressing levels of riO-1 mRNA and protein. As HO-1 expression in stably transfected cell lines altered, we use Western blot to test transfected cell lines P21, P27 protein expression levels. As result shows, we got 1 HO-over-expression of wild-type and mutant in HepG2 cells; wild- type and mutant＇s over expression of HO-1 can induce the expression of tumor suppressor genes p21 and p27.we got the conclusion that HO-l＇s over-expression of tumor suppressor genes p21 and p27 is unrelated to the expression of heme decomposition products. HO-1 may regulate the expression of p21 and p27 through other mechanisms.

Plant Anthocyanin Synthesis and Gene Regulation

This paper aims to explain the biochemistry of anthocyanin synthesis based on an overview of plant anthocyanin synthesis genes and environmental factors in the regulation of anthocyanin metabolism. The results show that： ① The metabolism of anthocyanins in plants is affected by the temperature, light, ultraviolet, fertilization status, hormone levels and other factors, which affect the military anthocyanin biosynthetic genes, and then induce or inhibit the synthesis of anthocyanins. ② In the regulation of genes, some of the structural genes of anthocyanin synthesis showed promoting effect, while others showed inhibitory effect. At different environ- mental conditions, the regulation of gene activation and inhibition of the amount of different regulatory genes that anthocyanin accumulation is different, and cause different colors of plant-organs production. ③ In different environmental factors or hor-mones induced to produce the same or different regulation of gene expression changes in regulatory genes, resulting in several different anthocyanins or anthocyanin ratio changes, so that the color of plant organs in different colors.

The Effect of Nitric Oxide on the Growth of Marine Phytoplankton

The incubation experiments of Skeletonema costatum, Dicrateria zhanjiangensis nov. sp., and Platymonas sub-cordiformis, and those of Emiliania huxleyi were carried out in the Marine Physical Chemistry Laboratory in Ocean University of China and in the Marine Organic Geochemistry Laboratory in the University of Georgia respectively. Nitric oxide was added into the media when these marine microalgae were growing. We found the growth of these four microalgae were promoted or inhibited when nitric oxide of different concentrations was added one or two times each day during the cultivation process. The results are consistent with the influence of nitric oxide on the growth of high plants. The results show that nitric oxide may be a new factor of regulation and control for the phytoplankton growth in seawater.

Observation of Transcription Regulation in the Mouse Heart Nuclear DNA Fragments and the Specific-protein Interaction by AFM

Using atom force microscopy (AFM), in vitro transcription, PAGE and other experimental technologies, it is observed that, in active genes of mice (Balb/c) nuclear DNA fragments of non-transcriptional state, only regulation sequences at both ends are associated with scaffold proteins (indissociable proteins) and some transcriptional factors such as complexes (dissociable proteins) made of gene-coding proteins and specific auxiliary small molecules, while there are no combining proteins in intermediate coding sequences. However, in active genes of transcriptional state, both regulation sequences and intermediate coding sequences are associated with active transcriptional factors by non-covalent bonds.This paper shows the prospective application of AFM observation and in vitro transcription in the research on gene expression and regulation. It also offers some theoretical basis for localization of specific genes in human genomes.

Specific effects of c-Jun NH2-terminal kinaseinteracting protein 1 in neuronal axons

c-Jun NH2-terminal kinase（JNK）-interacting protein 3 plays an important role in brain-derived neurotrophic factor/tropomyosin-related kinase B（Trk B） anterograde axonal transport. It remains unclear whether JNK-interacting protein 1 mediates similar effects, or whether JNK-interacting protein 1 affects the regulation of Trk B anterograde axonal transport. In this study, we isolated rat embryonic hippocampus and cultured hippocampal neurons in vitro. Coimmunoprecipitation results demonstrated that JNK-interacting protein 1 formed Trk B complexes in vitro and in vivo. Immunocytochemistry results showed that when JNK-interacting protein 1 was highly expressed, the distribution of Trk B gradually increased in axon terminals. However, the distribution of Trk B reduced in axon terminals after knocking out JNK-interacting protein 1. In addition, there were differences in distribution of Trk B after JNK-interacting protein 1 was knocked out compared with not. However, knockout of JNK-interacting protein 1 did not affect the distribution of Trk B in dendrites. These findings confirm that JNK-interacting protein 1 can interact with Trk B in neuronal cells, and can regulate the transport of Trk B in axons, but not in dendrites.

Effect of trans-acting factor on rat glutathione S-transferase P1 gene transcription regulation in tumor cells

Objective To investigate the effect of trans-acting factor(s) on rat glutathione S-transferase P1 gene (rGSTP1) transcription regulation in tumor cells.Methods The binding of trans-acting factor(s) to two enhancers of the rGSTP1 gene, glutathione S-transferase P enhancer Ⅰ (GPEI) and glutathione S-transferase P enhancer Ⅱ-1 (GPEⅡ-1), was identified by an electrophoretic mobility shift assay (EMSA). The molecular weight of trans-acting factor was measured in a UV cross-linking experiment. Results Trans-acting factor interacting with the core sequence of GPEI (cGPEI) were found in human cervical adenocarcinoma cell line (HeLa) and rat hepatoma cell line (CBRH7919). These proteins were not expressed in normal rat liver. Although specific binding proteins that bound to GPEⅡ-1 were detected in all three cell types, a 64 kDa binding protein that exists in HeLa and CBRH7919 cells was absent in normal rat liver. Conclusion cGPEI, GPEII specific binding proteins expressed in HeLa and CBRH7919 cells may play an important role in the high transcriptional level of the rGSTP1 gene in tumor cells.

Insights into the regulation of C-repeat binding factors in plant cold signaling

Cold temperatures, a major abiotic stress, threaten the growth and development of plants, worldwide. To cope with this adverse environmental cue, plants from temperate climates have evolved an array of sophisticated mechanisms to acclimate to cold periods, increasing their ability to tolerate freezing stress. Over the last decade, significant progress has been made in determining the molecular mechanisms underpinning cold acclimation, including following the identification of several pivotal components, including candidates for cold sensors, protein kinases, and transcription factors. With these developments, we have a better understanding of the CBF-dependent cold-signaling pathway. In this review, we summarize recent progress made in elucidating the cold-signaling pathways, especially the C-repeat binding factor-dependent pathway, and describe the regulatory function of the crucial components of plant cold signaling. We also discuss the unsolved questions that should be the focus of future work.

Negative regulation of resistance protein-mediated immunity by master transcription factors SARD_1 and CBP60g

Summary Salicylic acid （SA） is an essential defence hormone in plants. Upon pathogen infection, induced biosynthesis of SA is mediated by Isochorismate synthase 1 （ICS1）, whose gene transcription is controlled mainly through two redundant transcription factors, SAR Deficient 1 （SARD0 and Calmodulin- binding protein 6o-like g （CBP60g）.

Chromatin-remodeling factor OsINO80 is involved in regulation of gibberellin biosynthesis and is crucial for rice plant growth and development

The phytohormone gibberellin（GA） plays essential roles in plant growth and development. Here,we report that OsINO80, a conserved ATP-dependent chromatin-remodeling factor in rice（Oryza sativa）, functions in both GA biosynthesis and diverse biological processes. OsINO80-knockdown mutants, derived from either T-DNA insertion or RNA interference, display typical GA-deficient phenotypes, including dwarfism, reduced cell length, late flowering, retarded seed germination and impaired reproductive development. Consistently, transcriptome analyses reveal that OsINO80 knockdown results in downregulation by more than two-fold of over 1,000 genes, including the GA biosynthesis genes CPS_1 and GA_3ox_2, and the dwarf phenotype of OsINO80-knockdown mutants can be rescued by the application of exogenous GA3. Chromatin immunoprecipitation（Ch IP） experiments show that OsINO80 directly binds to the chromatin of CPS1 and GA_3ox_2 loci. Biochemical assays establish that OsINO80 specially interacts with histone variant H_2A.Z and the H_2A.Z enrichments at CPS_1 and GA_3ox_2 are decreased in OsINO80-knockdown mutants. Thus, our study identified a rice chromatin-remodeling factor,OsINO80, and demonstrated that OsINO80 is involved in regulation of the GA biosynthesis pathway and plays critical functions for many aspects of rice plant growth and development.

Molecular mechanism of microRNA125 regulating human coagulation factor IX gene with nonsense mutation

Objective To construct human coagulation factorⅨmini-gene(Mini-h F9)and some nonsense mutants,detect the levels of the Mini-h F9 mRNA,and analyze the molecular mechanism of microRNA125 regulating F9gene with nonsense mutation.Methods Three nonsense mutants were obtained by using PCR mutagenesis to ana-

中国近20年植被覆盖变化及其驱动机理

Vegetation change is of significant concern because it plays a crucial role in the global carbon cycle and climate.Many studies have examined recent changes in vegetation growth and the associated drivers.These drivers include both natural and human activities,but few studies have identified the regulation factors.By employing normalized difference vegetation index(NDVI)data,we analyzed the spatiotemporal pattern of vegetation change in China and then explored the driving factors.It was found that the overall greening of China has improved significantly,especially in the Loess Plateau and southwest China.The Yangtze River Delta and Bohai Rim,however,have not seen as much growth.Natural conditions are conducive to vegetation growth.Although socioeconomic development will be more beneficial for vegetation restoration,the current level and speed of development have a negative effect on vegetation.The regulation factors are considered separately since they affect both directly and indirectly.Regulation factors have accelerated vegetation growth.By understanding the factors affecting the current vegetation growth,we can provide a guide for future vegetation recovery in China and other similar countries.

Zhizhu Decoction Alleviates Intestinal Barrier Damage via Regulating SIRT1/FoxO1 Signaling Pathway in Slow Transit Constipation Model Mice

Objective:To explore the possible effects and mechanism of Zhizhu Decoction(ZZD)on the pathophysiology of slow transit constipation(STC).Methods A total of 54 C57BL/6 mice was randomly divided into the following 6 groups by a random number table,including control,STC model(model),positive control,and low-,medium-and high-doses ZZD treatment groups(5,10,20 g/kg,namely L,M-,and H-ZZD,respectively),9 mice in each group.Following 2-week treatment,intestinal transport rate(ITR)and fecal water content were determined,and blood and colon tissue samples were collected.Hematoxylin-eosin and periodic acid-Schiff staining were performed to evaluate the morphology of colon tissues and calculate the number of goblet cells.To determine intestinal permeability,serum levels of lipopolysaccharide(LPS),low-density lipoprotein(LDL)and mannose were measured using enzyme-linked immunosorbent assay(ELISA).Western blot analysis was carried out to detect the expression levels of intestinal tight junction proteins zona-occludens-1(ZO-1),claudin-1,occludin and recombinant mucin 2(MUC2).The mRNA expression levels of inflammatory cytokines including tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,IL-4,IL-10 and IL-22 were determined using reverse transcription-quantitative reverse transcription reaction.Colon indexes of oxidative stress were measured by ELISA,and protein expression levels of colon silent information regulator 1/forkhead box O transcription factor 1(SIRT1/FoxO1)antioxidant signaling pathway were detected by Western blot.Results Compared with the model group,ITR and fecal moisture were significantly enhanced in STC mice in the M-ZZD and H-ZZD groups(P<0.01).Additionally,ZZD treatment notably increased the thickness of mucosal and muscular tissue,elevated the number of goblet cells in the colon of STC mice,reduced the secretion levels of LPS,LDL and mannose,and upregulated ZO-1,claudin-1,occludin and MUC2 expressions in the colon in a dose-dependent manner,compared with the model group(P<0.05 or P<0.01).In addition,ZZD significantly attenuated intestinal inflammation and oxidative stress and activated the SIRT1/FoxO1 signaling pathway(P<0.05 or P<0.01).Conclusion ZZD exhibited beneficial effects on the intestinal system of STC mice and alleviated intestinal inflammation and oxidative stress via activating SIRT1/FoxO1 antioxidant signaling pathway in the colon.

A high-resolution cell atlas of the domestic pig lung and an online platform for exploring lung single-cell data

The genetically engineered pig is regarded as an optimal source of organ transplantation for humans and an excellent model for human disease research,given its comparable physiology to human beings.A myriad of single-cell RNA sequencing(sc RNA-seq)data on humans has been reported,but such data on pigs are scarce.Here,we apply sc RNA-seq technology to study the cellular heterogeneity of 3-month-old pig lungs,generating the single-cell atlas of 13,580 cells covering 16 major cell types.Based on these data,we systematically characterize the similarities and differences in the cellular cross-talk and expression patterns of respiratory virus receptors in each cell type of pig lungs compared with human lungs.Furthermore,we analyze pig lung xenotransplantation barriers and reported the cell-type expression patterns of 10 genes associated with pig-to-human immunobiological incompatibility and coagulation dysregulation.We also investigate the conserved transcription factors(TFs)and their candidate target genes and constructed five conserved TF regulatory networks in the main cell types shared by pig and human lungs.Finally,we present a comprehensive and openly accessible online platform,Scdb Lung.Our sc RNA-seq atlas of the domestic pig lung and Scdb Lung database can guide pig lung research and clinical applicability.

Inhibition Mechanism of Qingluo Tongbi Granule(清络通痹颗粒)on Osteoclast Differentiation Induced by Synovial Fibroblast and Monocytes Co-Culture in Adjuvant-Induced Arthritic Rats

Objective： To study the mechanism underlying the inhibitory effect of Qingluo Tongbi Granule （清络通痹颗粒, QTG） on osteoclast differentiation in rheumatoid arthritis in rats. Methods： Fibroblast and monocyte co-culture were used to induce osteoclast differentiation in adjuvant-induced arthritic （AIA） rats. Serum containing QTG was prepared and added to the osteoclasts, and activation of the tumor necrosis factor receptorassociated factor 6/mitogen-activated protein kinase/nuclear factor of activated T cells, cytoplasmic1 （TRAF6/ MAPK/NFATcl） pathways was examined. Results： The induced osteoclasts were multinucleated and stained positive for tartrate-resistant acid phosphatase （TRAP） staining. Serum containing QTG at 14.4, 7.2 or 3.6 g/kg inhibited the activation of TRAF6, extracellular regulated protein kinase （ERK）1/2, c-Jun N-terminal kinase （JNK） and p38 and decreased the percentage of cells with nuclear NFATcl in a dose-dependent manner, the high and middle doses exhibited clear inhibitory activity （P〈0.01 and P〈0.05, respectively）. After the addition of MAPK inhibitors, the NFATcl expression showed no significant difference compared with the control group （P〉0.05）. Conclusions： Serum containing QTG could generally inhibit the TRAF6/MAPK pathways and possibly inhibit the NFATcl pathway. In addition, QTG may regulate other signaling pathways that are related to osteoclast differentiation and maturation.

Panax notoginseng Saponins Protect Kidney from Diabetes by Up-regulating Silent Information Regulator 1 and Activating Antioxidant Proteins in Rats

Objective： To explore the mechanism of the protective effects of Panax notoginseng saponins（PNS） on kidney in diabetic rats. Methods： Diabetic rat model was obtained by intravenous injection of alloxan, and the rats were divided into model, PNS-100 mg/（kg·day） and PNS-200 mg/（kg·day） groups, 10 each. Another 10 rats injected with saline were served as control. Periodic acid-Schiff staining and immunological histological chemistry were used to observe histomorphology and tissue expression of bone morphogenetic protein-7（BMP-7）. Silent information regulator 1（SIRT1） was silenced in rat mesangial cells by RNA interference. The mR NA expressions of SIRT-1, monocyte chemoattractant protein-1（MCP-1）, transforming growth factor β1（TGF-β1） and plasminogen activator inhibitor-1（PAI-1） were analyzed by reverse transcription polymerase chain reaction. The protein expressions of SIRT1 and the acetylation of nuclear factor κB（NF-κB） P65 were determined by western blotting. The concentration of MCP-1, TGF-β1 and malondialdehyde（MDA） in culture supernatant were detected by enzyme-linked immuno sorbent assay. The activity of superoxide dismutase（SOD） was detected by the classical method of nitrogen and blue four. Results： In diabetic model rats, PNS could not only reduce blood glucose and lipid（P〈0.01）, but also increase protein level of BMP-7 and inhibit PAI-1 expression for suppressing fibrosis of the kidney. In rat mesangial cells, PNS could up-regulate the expression of SIRT1（P〈0.01） and in turn suppress the transcription of TGF-β1（P〈0.05） and MCP-1（P〈0.05）. PNS could also reverse the increased acetylation of NF-κB p65 by high glucose. In addition, redox regulation factor MDA was down-regulated（P〈0.05） and SOD was up-regulated（P〈0.01）, which were both induced by SIRT1 up-regulation. Conclusions： PNS could protect kidney from diabetes with the possible mechanism of up-regulating SIRT1, therefore inhibiting inflammation through decreasing the induction of inflammatory cytokines and TGF-β1, as well as activating antioxidant proteins.

A new perspective on ecological vulnerability and its transformation mechanisms

Ecological vulnerability refers to the degree of ecosystem disturbance,system damage,and the ability of system restoration.Although case-specific evaluations of ecological vulnerability are progressing rapidly,they have been carried out mainly in areas with intensive human activities or in harsh natural environments.Using the Web of ScienceTM core collection,this review paper summarized studies on ecological vulnerability published from 2000 to 2022 and analyzed in depth major case studies.It was found that traditional ecological vulnerability research has been addressed largely in terms of assessment models,data processing models,and analysis of influencing factors,however there was a lack of research on the process of vulnerability transformation.Because the vulnerability transformation in a hierarchical vulnerability index system is regulated by multiple factors in a heterogeneous region,it is urgent to understand how the ecological vulnerability in a region evolves from one level to another over time.Therefore this paper put forward the new perspective of research,i.e.,applying quantitative analysis to identification of regulating factors and exploring the mechanisms of ecological vulnerability transformation.This new perspective could assist in monitoring the complex spatiotemporal changes in ecological vulnerability and taking necessary measures to prevent from decline of ecological stability.