Insight into Function and Subcellular Localization of a Type III-Secreted Effector in Pseudomonas syringae pv. tomato DC3000

Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) is a bacterial pathogen of tomato and of the model plants Arabidopsis and Nicotiana benthamiana (N. benthamiana). Like numerous Gram-negative bacterial pathogens of animals and plants, Pst DC3000 exploits the conserved type III secretion system (TTSS) to deliver multiple virulence effectors directly into the host cells. Type III effectors (T3Es) collectively participate in causing disease, by mechanisms that are not well clarity. Elucidating the virulence function of individual effector is fundamental for understanding bacterial infection of plants. Here, we focused on studying one of these effectors, HopAA1-1, and analyzed its potential function and subcellular localization in N. benthamiana. Using an Agrobacterium-mediated transient expression system, we found that HopAA1-1 can trigger domain-dependent cell death in N. benthamiana. The observation using confocal microscopy showed that the YFP-tagged HopAA1-1 localizes to diverse cellular components containing nucleus, cytoplasm and cell membrane, which was demonstrated through immunoblot analysis of membrane fractionation and nuclear separation. Enforced HopAA1-1 subcellular localization, by tagging with a nuclear localization sequence (NLS) or a nuclear export sequence (NES), shows that HopAA1-1-induced cell death in N. benthamiana is suppressed in the nucleus but enhanced in the cytoplasm. Our research is lay a foundation for revealed the molecular pathogenesis of Pseudomonas syringae pv. tomato.

The Application of Nicotiana benthamiana as a Transient Expression Host to Clone the Coding Sequences of Plant Genes

Coding sequences (CDS) are commonly used for transient gene expression, in yeast two-hybrid screening, to verify protein interactions and in prokaryotic gene expression studies. CDS are most commonly obtained using complementary DNA (cDNA) derived from messenger RNA (mRNA) extracted from plant tissues and generated by reverse transcription. However, some CDS are difficult to acquire through this process as they are expressed at extremely low levels or have specific spatial and/or temporal expression patterns in vivo. These challenges require the development of alternative CDS cloning technologies. In this study, we found that the genomic intron-containing gene coding sequences (gDNA) from Arabidopsis thaliana, Oryza sativa, Brassica napus, and Glycine max can be correctly transcribed and spliced into mRNA in Nicotiana benthamiana. In contrast, gDNAs from Triticum aestivum and Sorghum bicolor did not function correctly. In transient expression experiments, the target DNA sequence is driven by a constitutive promoter. Theoretically, a sufficient amount of mRNA can be extracted from the N. benthamiana leaves, making it conducive to the cloning of CDS target genes. Our data demonstrate that N. benthamiana can be used as an effective host for the cloning CDS of plant genes.

Screening and Identifying of Interaction Protein AtL5 in Arabidopsis thaliana

Research background: The Arabidopsis-resistance protein L5 (AT1G12290) can trigger cell death in Nicotiana benthamiana, which is a characteristic function of an NBS-LRR (Nucleotide-Binding Sites and Leucine-Rich Repeat) protein activation. Purpose: To explore the function and molecular regulatory network of L5. Method: We employed yeast two-hybrid technology to search for interacting proteins of L5, combined with laser confocal microscopy to observe the subcellular localization of these candidate proteins, and analyzed the impact of these proteins on L5 function using an Agrobacterium mediated transient expression system. Results: Seven candidate interacting proteins were identified from the Arabidopsis cDNA library, including PPA1 (AT1G01050), RIN4 (AT3G25070), LSU1 (AT3G49580), BZIP24 (AT3G51960), BOI (AT4G19700), RING/U (AT4G22250) and PPA3 (AT2G46860). Functional analysis of these candidate interacting proteins showed that they participated in multiple pathways, including biological and abiotic stress, programmed cell death, protein degradation, material metabolism and transcriptional regulation. The results of laser confocal microscopy manifested that RIN4 was only localized on the plasma membrane (PM), and RING/U was mainly associated with the PM. PPA1, PPA3, LSU1, BZIP24, and BOI all emerged nuclear and cytoplasmic localization. The results of the transient assay proclaimed that both BOI and RING/U can inhibit cell death caused by L5. Conclusions: These results indicate that L5 immune receptors may participate in various pathways, and their protein levels and activities are strictly regulated at multiple levels, providing a basis for elucidating the mechanism of L5 immune receptors in Arabidopsis resistance.

Expression Pattern, Interaction Network, and Functional Analysis of the Arabidopsis Botrytis Susceptible1 Interactor

E3 ubiquitin ligases are participated in numerous processes, regulating the response to biotic and abiotic stresses. Botrytis susceptible1 interactor (BOI) is a RING (Really Interesting New Gene)-type E3 ligase that mediates the ubiquitination of BOS1 (Botrytis susceptible1), a transcription factor involved in stress and pathogen responses. Although BOI is an E3 ligase, there are reports to show that BOI interacts with target proteins such as DELLAs or CONSTANS to repress gibberellin responses and flowering without the degradation of the target proteins. In this article, we utilize diversified methods to comprehensively analyze the expression pattern, interaction network and function of BOI gene. Firstly, 1800 bp upstream region of BOI gene from Arabidopsis thaliana (Arabidopsis) genome was isolated, and fused GUS reporter gene. The resulting expression cassette was introduced into wild-type Arabidopsis through Agrobacterium-mediated transformation. The result demonstrated that BOI gene was expressed predominantly in leaves, siliques, young roots, and flowering tissues, indicating that BOI gene may be involved in multiple processes in plant growth and development in Arabidopsis. Besides, eight candidate interacting proteins were obtained from the Arabidopsis cDNA library via yeast two-hybrid technology, including EXO70E2 (AT5G61010), WRKY7 (AT4G24240), WRKY11 (AT4G31550), WRKY17 (AT2G24570), UBP20 (AT4G17895), L5 (AT1G12290), SAUR9 (AT4G36110) and TCP21 (AT5G08330). Functional analysis of these candidate interacting proteins manifested that they related to multiple pathways, including biological and abiotic stress, programmed cell death, protein degradation, material metabolism and transcriptional regulation. In addition, the results of the transient assay proclaimed that BOI protein affects the protein stability of EXO70E2 and L5 through its E3 ubiquitin ligase activity. Our results provide novel clues for a better understanding of molecular mechanisms underlying BOI-mediated regulations.

Non-alcoholic fatty liver disease and sleep disorders

Studies have shown that non-alcoholic fatty liver disease(NAFLD)may be associated with sleep disorders.In order to explore the explicit relationship between the two,we systematically reviewed the effects of sleep disorders,especially obstructive sleep apnea(OSA),on the incidence of NAFLD,and analyzed the possible mechanisms after adjusting for confounding factors.NAFLD is independently associated with sleep disorders.Different sleep disorders may be the cause of the onset and aggravation of NAFLD.An excessive or insufficient sleep duration,poor sleep quality,insomnia,sleep-wake disorders,and OSA may increase the incidence of NAFLD.Despite that some research suggests a unidirectional causal link between the two,specifically,the onset of NAFLD is identified as a result of changes in sleep characteristics,and the reverse relationship does not hold true.Nevertheless,there is still a lack of specific research elucidating the reasons behind the higher risk of developing sleep disorders in individuals with NAFLD.Further research is needed to establish a clear relationship between NAFLD and sleep disorders.This will lay the groundwork for earlier identification of potential patients,which is crucial for earlier monitoring,diagnosis,effective prevention,and treatment of NAFLD.

Structural and Functional Insights into an Arabidopsis NBS-LRR Receptor in Nicotiana benthamiana

Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involved in autoimmunity initiated by type one protein phosphatase 4 mutation (topp4-1) in Arabidopsis, however, its role in planta is still unclear. This study employed Nicotiana benthamiana, a model platform, to conduct an overall structural and functional analysis of SUT1 protein. The transient expression results revealed that SUT1 is a typical CNL (CC-NBS-LRR) receptor, both fluorescence data and biochemical results showed the protein is mainly anchored on the plasma membrane due to its N-terminal acylation site. Further truncation experiments announced that its CC (coiled-coil) domain possessed cell-death-inducing activity. The outcomes of point mutations analysis revealed that not only the CC domain, but also the full-length SUT1 protein, whose function and subcellular localization are influenced by highly conserved hydrophobic residues. These research outcomes provided favorable clues for elucidating the activation mechanism of SUT1.

Dissecting Multiple Arabidopsis CC-NBS-LRR Proteins Structure and Localization

NBS-LRR (nucleotide binding sites and leucine rich repeat) protein plays a crucial role as sentries and as defense activators in plants. The structure and function of NBS-LRR proteins are closely related. Previous articles have announced that the activated ZAR1 (HopZ-Activated Resistance 1) forms a pentamer in the plasma membrane, which is a calcium permeable channel that can trigger plant immune signaling and cell death. However, the structure of galore NBS-LRRs in Arabidopsis is not yet clear. The functional sites of distinct NBS-LRR in cells may vary. In addition, identifying pathogens and activating defense regions may occur in different subcellular compartments. Therefore, dissecting the specific structure and positioning of NBS-LRRs is an indispensable step in understanding their functions. In this article, we exploit AlphaFold to predict the structure of some designed NBS-LRRs, and utilize Agroinfiltration transient expression system, combined with biochemical fractionation, to dissect the localization of these NBS-LRR receptors from Arabidopsis. Structural data indicates that the identified NBS-LRRs share analogous conformation. Membrane fractionation assay demonstrates these NBS-LRRs are mainly associated with the membrane. These data show that the Ca2+-permeable channel activity may be evolutionarily conserved in NBS-LRR of Arabidopsis, and this study provides some reference clues for analyzing the structure and localization patterns of other plant immune receptors.

Integration of artificial intelligence and multi-omics in kidney diseases

Kidney disease is a leading cause of death worldwide.Currently,the diagnosis of kidney diseases and the grading of their severity are mainly based on clinical features,which do not reveal the underlying molecular pathways.More recent surge of∼omics studies has greatly catalyzed disease research.The advent of artificial intelligence(AI)has opened the avenue for the efficient integration and interpretation of big datasets for discovering clinically actionable knowledge.This review discusses how AI and multi-omics can be applied and integrated,to offer opportunities to develop novel diagnostic and therapeutic means in kidney diseases.The combination of new technology and novel analysis pipelines can lead to breakthroughs in expanding our understanding of disease pathogenesis,shedding new light on biomarkers and disease classification,as well as providing possibilities of precise treatment.

Hemodialysis bilayer bionic blood vessels developed by the mechanical stimulation of hepatitis B viral X(HBX)genetransfected hepatic stellate cells

Artificial vascular graft(AVG)fistula is widely used for hemodialysis treatment in patients with renal failure.However,it has poor elasticity and compliance,leading to stenosis and thrombosis.The ideal artificial blood vessel for dialysis should replicate the structure and components of a real artery,which is primarily maintained by collagen in the extracellular matrix(ECM)of arterial cells.Studies have revealed that in hepatitis B virus(HBV)-induced liver fibrosis,hepatic stellate cells(HSCs)become hyperactive and produce excessive ECM fibers.Furthermore,mechanical stimulation can encourage ECM secretion and remodeling of a fiber structure.Based on the above factors,we transfected HSCs with the hepatitis B viral X(HBX)gene for simulating the process of HBV infection.Subsequently,these HBX-HSCs were implanted into a polycaprolactonepolyurethane(PCL-PU)bilayer scaffold in which the inner layer is dense and the outer layer consists of pores,which was mechanically stimulated to promote the secretion of collagen nanofiber from the HBX-HSCs and to facilitate crosslinking with the scaffold.We obtained an ECM-PCL-PU composite bionic blood vessel that could act as access for dialysis after decellularization.Then,the vessel scaffold was implanted into a rabbit’s neck arteriovenous fistula model.It exhibited strong tensile strength and smooth blood flow and formed autologous blood vessels in the rabbit’s body.Our study demonstrates the use of human cells to create biomimetic dialysis blood vessels,providing a novel approach for creating clinical vascular access for dialysis.

Virtual Screening and Mechanism Analysis of Effective Components from Several Chinese Herbs to Inhibit dPLA2 of Deinagkistrodon acutus Venom

Objective: In folk and TCM clinical medicine, Chinese herbal medicine is used to treat snakebite and has good curative effect, but its active ingredients and mechanism are still unclear. In this study, virtual screening and mechanism analysis of effective components from 6 Chinese herbs to inhibit phospholipase A2 of Deinagkistrodon acutus (dPLA2) venom were conducted. Methods: With advanced computing software AutoDock, Pymol and GROMACS, the molecules selected from the Chinese herbal Medicine Chemical Composition databas6e (TCMSP) were docked with the dPLA2 from the protein database (PDB). Further molecular dynamics simulation was used to evaluate the molecular binding stability. Results: Four potential dPLA2-inhibiting molecules were screened: lobelanidine, lobeline, norlobelanine and pratensein, by analyzing the spatial structure, binding energy and binding interaction of small molecular-dPLA2 complexes, as well as the RMSD and RMSF of molecular dynamics simulation. Conclusion: To our knowledge, this is the first report of lobeline has an inhibitory effect on dPLA2, and lobelanidine, as a precursor of lobeline, has a stronger inhibitory effect. According to the docking results, it is speculated that the mechanism of action of the four molecules is to form stable interactions with calcium ions and amino acid residues on the calcium ion binding ring in dPLA2. Moreover, these small molecules compete with phosphatidylcholine (the natural substrate of dPLA2) to bind dPLA2 and have a higher affinity than phosphatidylcholine, resulting in inhibition of dPLA2 activity.

Chinese cohort study of chronic kidney disease： design and methods

Background Chronic kidney disease （CKD） is a common disorder associated with multiple adverse clinical consequences,especially cardiovascular risk and end-stage renal disease.A recent national survey demonstrated that CKD has become a leading health problem in China.There is an urgent need to implement an in-depth investigation of the CKD burden and also to explore underlying mechanisms of CKD progression and it association with adverse consequences.Methods The Chinese Cohort Study of Chronic Kidney Disease （C-STRIDE） is the first national CKD cohort in China.It will enroll approximately 3 000 pre-dialysis CKD patients aged between 18 and 74 years and follow-up for at least 5 years.Questionnaires,anthropometric measures,laboratory tests,and biomaterials will be collected at baseline and annually.The principal clinical outcomes of the C-STRIDE consist of renal disease events,cardiovascular events,and death.Based on the longitudinal clinical data and biomaterials,the risk factors with CKD progression and other outcomes will be analyzed,and candidate markers and predicted models will be established.Conclusion The C-STRIDE would provide important evidence for underlying mechanisms of CKD progression,valuable information for clinical guidelines,and healthcare policies in China.

Lanthanum promoting bone formation by regulating osteogenesis,osteoclastogenesis and angiogenesis

With lanthanum(La) in agriculture, medicine, and the chemical industry, La shows accumulation in the body, especially in the bone, because of its similar structure to calcium. Moreover, La has a direct role in bone formation regulating osteoblasts and osteoclasts. Nevertheless, bone formation is complex under the regulation of osteogenesis, osteoclastogenesis, and angiogenesis in the bone microenvironment. It is difficult to comprehensively understand the regulation of bone homeostasis from a single kind of cell.Herein, some bone microenvironment-related cells and mouse calvaria culture models were used to study the regulatory effect of La-based compound-lanthanum nitrate(La(NO_(3))_(3)) on bone formation.La(NO_(3))_(3) has good biological safety, and the osteogenic differentiation is significantly increased under La(NO_(3))_(3) treatment for bone marrow mesenchymal stem cells(BMMSC). In contrast, the differentiation,maturation, and bone erosion ability of osteoclasts are significantly decreased. Meanwhile, the angiogenesis ability of human umbilical vein endothelial cells(HUVEC) is significantly promoted when treated with La(NO_(3))_(3). Furthermore, bone metabolism and angiogenesis are improved under La(NO_(3))_(3) treatment in the calvaria ex vivo culture model and BMMSC-HUVEC co-culture system. These results suggest that La has the beneficial effect of promoting bone metabolism and improving bone formation in the bone microenvironment.

An orally administered bacterial membrane protein nanodrug ameliorates doxorubicin cardiotoxicity through alleviating impaired intestinal barrier

The cardiotoxicity caused by Dox chemotherapy represents a significant limitation to its clinical application and is a major cause of late death in patients undergoing chemotherapy.Currently,there are no effective treatments available.Our analysis of 295 clinical samples from 132 chemotherapy patients and 163 individuals undergoing physical examination revealed a strong positive correlation between intestinal barrier injury and the development of cardiotoxicity in chemotherapy patients.We developed a novel orally available and intestinal targeting protein nanodrug by assembling membrane protein Amuc_1100(obtained from intestinal bacteria Akkermansia muciniphila),fluorinated polyetherimide,and hyaluronic acid.The protein nanodrug demonstrated favorable stability against hydrolysis compared with free Amuc_1100.The in vivo results demonstrated that the protein nanodrug can alleviate Dox-induced cardiac toxicity by improving gut microbiota,increasing the proportion of short-chain fatty acid-producing bacteria from the Lachnospiraceae family,and further enhancing the levels of butyrate and pentanoic acids,ultimately regulating the homeostasis repair of lymphocytes in the spleen and heart.Therefore,we believe that the integrity of the intestinal barrier plays an important role in the development of chemotherapy-induced cardiotoxicity.Protective interventions targeting the intestinal barrier may hold promise as a general clinical treatment regimen for reducing Dox-induced cardiotoxicity.

Polysaccharides from Chinese herbal medicine:a review on the hepatoprotective and molecular mechanism

Polysaccharides,predominantly extracted from traditional Chinese medicinal herbs such as Lycium barbarum,Angelica sinensis,Astragalus membranaceus,Dendrobium officinale,Ganoderma lucidum,and Poria cocos,represent principal bioactive constituents extensively utilized in Chinese medicine.These compounds have demonstrated significant anti-inflammatory capabilities,especially anti-liver injury activities,while exhibiting minimal adverse effects.This review summarized recent studies to elucidate the hepatoprotective efficacy and underlying molecular mechanisms of these herbal polysaccharides.It underscored the role of these polysaccharides in regulating hepatic function,enhancing immunological responses,and improving antioxidant capacities,thus contributing to the attenuation of hepatocyte apoptosis and liver protection.Analyses of molecular pathways in these studies revealed the intricate and indispensable functions of traditional Chinese herbal polysaccharides in liver injury management.Therefore,this review provides a thorough examination of the hepatoprotective attributes and molecular mechanisms of these medicinal polysaccharides,thereby offering valuable insights for the advancement of polysaccharide-based therapeutic research and their potential clinical applications in liver disease treatment.

A wearable,rapidly manufacturable,stabilityenhancing microneedle patch for closed-loop diabetes management

The development of a wearable,easy-to-fabricate,and stable intelligent minisystem is highly desired for the closedloop management of diabetes.Conventional systems always suffer from large size,high cost,low stability,or complex fabrication.Here,we show for the first time a wearable,rapidly manufacturable,stability-enhancing microneedle patch for diabetes management.The patch consists of a graphene composite ink-printed sensor on hollow microneedles,a polyethylene glycol(PEG)-functionalized electroosmotic micropump integrated with the microneedles,and a printed circuit board for precise and intelligent control of the sensor and pump to detect interstitial glucose and deliver insulin through the hollow channels.Via synthesizing and printing the graphene composite ink,the sensor fabrication process is fast and the sensing electrodes are stable.The PEG functionalization enables the micropump a significantly higher stability in delivering insulin,extending its lifetime from days to weeks.The patch successfully demonstrated excellent blood glucose control in diabetic rats.This work may introduce a new paradigm for building new closedloop systems and shows great promise for widespread use in patients with diabetes.

Astragalus Polysaccharide Enhances Voriconazole Metabolism under Inflammatory Conditions through the Gut Microbiota

Background and Aims:Voriconazole(VRC),a widely used antifungal drug,often causes hepatotoxicity,which presents a significant clinical challenge.Previous studies demonstrated that Astragalus polysaccharide(APS)can regulate VRC metabolism,thereby potentially mitigating its hepatotoxic effects.In this study,we aimed to explore the mechanism by which APS regulates VRC metabolism.Methods:First,we assessed the association of abnormal VRC metabolism with hepatotoxicity using the Roussel Uclaf Causality Assessment Method scale.Second,we conducted a series of basic experiments to verify the promotive effect of APS on VRC metabolism.Various in vitro and in vivo assays,including cytokine profiling,immunohistochemistry,quantitative polymerase chain reaction,metabolite analysis,and drug concentration measurements,were performed using a lipopolysaccharideinduced rat inflammation model.Finally,experiments such as intestinal biodiversity analysis,intestinal clearance assessments,and Bifidobacterium bifidum replenishment were performed to examine the ability of B.bifidum to regulate the expression of the VRC-metabolizing enzyme CYP2C19 through the gut–liver axis.Results:The results indicated that APS does not have a direct effect on hepatocytes.However,the assessment of gut microbiota function revealed that APS significantly increases the abundance of B.bifidum,which could lead to an anti-inflammatory response in the liver and indirectly enhance VRC metabolism.The dual-luciferase reporter gene assay revealed that APS can hinder the secretion of pro-inflammatory mediators and reduce the inhibitory effect on CYP2C19 transcription through the nuclear factor-κB signaling pathway.Conclusions:The study offers valuable insights into the mechanism by which APS alleviates VRC-induced liver damage,highlighting its immunomodulatory influence on hepatic tissues and its indirect regulatory control of VRC-metabolizing enzymes within hepatocytes.

ncidence, Development, and Prognosis of Diabetic Kidney Disease in China： Design and Methods

Background： Although that glomerulonephritis is the major cause of end-stage renal disease in developing countries such as China, the increasing prevalence of diabetes has contributed to the changing spectrum of predialysis chronic kidney disease. Recent studies have revealed an increased proportion of patients with diabetic kidney disease （DKD） in hemodialysis populations in large cities in China. However, studies regarding the clinical phenotype of DKD in China are extremely limited. The incidence, development, and prognosis of diabetic kidney disease （INDEED） study aims to investigate the incidence, progression, and prognosis of DKD, as well as the associated genetic, behavioral, and environmental factors and biomarkers in patients with DKD in China. Methods： INDEED study is a prospective cohort study based on all participants with diabetes in the Kailuan study, which is a general population-based cohort study in northern China. Altogether, over 10,000 participants with diabetes will be followed biennially. Questionnaires documenting general characteristics, behavioral and environmental factors, and medical history will be administrated. Anthropometric measurements and a series of laboratory tests will be peribrmed in one central laboratory. The DNA, plasma, and urine samples of every participant will be stored in a biobank for future research. Conclusions： INDEED study will provide essential information regarding the clinical phenotype and prognosis of patients with DKD in China and will be valuable to identity factors and biomarkers associated with patients with DKD in China.

Expert consensus on the diagnosis and management of diabetic kidney disease

For the past decades,the rapid growth of economy and transition of lifestyles have led to a rapid rise in the prevalence of diabetes in China.Due to the lack of standard process in prevention and management of diabetic kidney disease(DKD)in China,the Peking University Health Science Center organized experts in fields of nephrology and endocrinology to develop the Chinese consensus statement of DKD.This consensus emphasizes on evidence-based inquiry,and focuses on the following three topics:(1)risk factors of the incidence and progression of DKD;(2)the diagnosis of DKD and indication of renal biopsy;(3)the overall management of DKD.

Homocysteine and IgA nephropathy: observational and Mendelian randomization analyses

Background:High levels of plasma homocysteine occur almost uniformly in patients with end-stage renal disease(ESRD).IgA nephropathy(IgAN)is the most common form of primary glomerulonephritis and a common cause of ESRD in young adults.Here,we aimed to detect whether homocysteine was elevated and associated with clinical-pathologic manifestations of IgAN patients and tested its causal effects using a two-sample Mendelian randomization(MR)approach.Methods:For observational analysis,108 IgAN patients,30 lupus nephritis(LN)patients,50 minimal change disease(MCD)patients,and 206 healthy controls were recruited from April 2014 to April 2015.Their plasma homocysteine was measured and clinical-pathologic manifestations were collected from medical records.For MR analysis,we further included 1686 IgAN patients.The missense variant methylenetetrahydrofolate reductase C677T(rs1801133)was selected as an instrument,which was genotyped by TaqMan allele discrimination assays.Results:Majority of IgAN patients(93.52%,101/108)showed elevated levels of plasma homocysteine(>10μmol/L).Plasma homocysteine in IgAN patients was significantly higher than that in MCD patients(median:18.32 vs.11.15μmol/L,Z=-5.29,P<0.01)and in healthy controls(median:18.32 vs.10.00μmol/L,Z=-8.76,P<0.01),but comparable with those in LN patients(median:18.32 L vs.14.50μmol/L,Z=-1.32,P=0.19).Significant differences were observed in sub-groups of IgAN patients according to quartiles of plasma homocysteine for male ratio(22.22%vs.51.85%vs.70.37%vs.70.37%,χ2=14.29,P<0.01),serum creatinine(median:77.00 vs.100.00 vs.129.00 vs.150.00μmol/L,χ2=34.06,P<0.01),estimated glomerular filtration rate(median:100.52 vs.74.23 vs.52.68 vs.42.67 mL·min-1·1.73 m-2,χ2=21.75,P<0.01),systolic blood pressure(median:120.00 vs.120.00 vs.125.00 vs.130.00 mmHg,χ2=2.97,P=0.05),diastolic blood pressure(median 80.00 vs.75.00 vs.80.00 vs.81.00 mmHg,χ2=11.47,P<0.01),and pathologic tubular atrophy and interstitial fibrosis(T)(T0/T1/T2:62.96%/33.33%/3.70%vs.29.63%/40.74%/29.63%vs.24.00%/48.00%/28.00%vs.14.81%/37.04%/48.15%,χ2=17.66,P<0.01).The coefficient of each rs1801133-T allele on homocysteine levels after controlling age and sex was 7.12(P<0.01).MR estimates showed causal positive effects of homocysteine on serum creatine(β=0.76,P=0.02),systolic blood pressure(β=0.26,P=0.02),diastolic blood pressure(β=0.20,P=0.01),and pathologic T lesion(β=0.01,P=0.01)in IgAN.Conclusions:By observational and MR analyses,consistent results were observed for associations of plasma homocysteine with serum creatinine,blood pressures,and pathologic T lesion in IgAN patients.

Relationship between Gut Microbiota and Phosphorus Metabolism in Hemodialysis Patients:A Preliminary Exploration

Background:Hyperphosphatemia is a risk factor associated with mortality in patients on maintenance hemodialysis.Gut absorption of phosphate is the major source.Recent studies indicated that the intestinal flora of uremic patients changed a lot compared with the healthy population,and phosphorus is an essential element of bacterial survival and reproduction.The purpose of this study was to explore the role of intestinal microbiota in phosphorus metabolism. Methods:A prospective self-control study was performed from October 2015 to January 2016.Microbial DNA was isolated from the stools of 20 healthy controls and 21 maintenance hemodialysis patients.Fourteen out of the 21 patients were treated with lanthanum carbonate for 12 weeks.Thus,stools were also collected before and after the treatment.The bacterial composition was analyzed based on 16S ribosomal RNA pyrosequencing.Bioinformatics tools,including sequence alignment,abundance profiling,and taxonomic diversity, were used in microbiome data analyses.Correlations between genera and the serum phosphorus were detected with Pearson's correlation. For visualization of the internal interactions and further measurement of the microbial community,SparCC was used to calculate the Spearman correlation coefficient with the corresponding P value between each two genera. Results:Thirteen genera closely correlated with serum phosphorus and the correlation coefficient was above 0.4(P <0.05).We also found that 58 bacterial operational taxonomic units (OTUs)were significantly different and more decreased OTUs were identified and seven genera (P <0.05)were obviously reduced after using the phosphate binder.Meanwhile,the microbial richness and diversity presented downward trend in hemodialysis patients compared with healthy controls and more downward trend after phosphorus reduction.The co-occurrence network of genera revealed that the network complexity of hemodialysis patients was significantly higher than that of controls,whereas treatment with lanthanum carbonate reduced the network complexity. Conclusions:Gut flora related to phosphorus metabolism in hemodialysis patients,and improving intestinal microbiota may regulate the absorption of phosphate in the intestine.The use of phosphate binder lanthanum carbonate leads to a tendency of decreasing microbial diversity and lower network complexity.