Altered Iron-Mediated Metabolic Homeostasis Governs the Efficacy and Toxicity of Tripterygium Glycosides Tablets Against Rheumatoid Arthritis

Rheumatoid arthritis(RA),a globally increasing autoimmune disorder,is associated with increased disability rates due to the disruption of iron metabolism.Tripterygium glycoside tablets(TGTs),a Tripterygium wilfordii Hook.f.(TwHF)-based therapy,exhibit satisfactory clinical efficacy for RA treatment.However,drug-induced liver injury(DILI)remains a critical issue that hinders the clinical application of TGTs,and the molecular mechanisms underlying the efficacy and toxicity of TGTs in RA have not been fully elucidated.To address this problem,we integrated clinical multi-omics data associated with the anti-RA efficacy and DILI of TGTs with the chemical and target profiling of TGTs to perform a systematic network analysis.Subsequently,we identified effective and toxic targets following experimental validation in a collagen-induced arthritis(CIA)mouse model.Significantly different transcriptome–protein–metabolite profiles distinguishing patients with favorable TGTs responses from those with poor outcomes were identified.Intriguingly,the clinical efficacy and DILI of TGTs against RA were associated with metabolic homeostasis between iron and bone and between iron and lipids,respectively.Particularly,the signal transducer and activator of transcription 3(STAT3)–hepcidin(HAMP)/lipocalin 2(LCN2)–tartrate-resis tant acid phosphatase type 5(ACP5)and STAT3–HAMP–acyl-CoA synthetase long-chain family member 4(ACSL4)–lysophosphatidylcholine acyltransferase 3(LPCAT3)axes were identified as key drivers of the efficacy and toxicity of TGTs.TGTs play dual roles in ameliorating CIA-induced pathology and in inducing hepatic dysfunction,disruption of lipid metabolism,and hepatic lipid peroxidation.Notably,TGTs effectively reversed“iron–bone”disruptions in the inflamed joint tissues of CIA mice by inhibiting the STAT3–HAMP/LCN2–ACP5 axis,subsequently leading to“iron–lipid”disturbances in the liver tissues via modulation of the STAT3–HAMP–ACSL4–LPCAT3 axis.Additional bidirectional validation experiments were conducted using MH7A and AML12 cells to confirm the bidirectional regulatory effects of TGTs on key targets.Collectively,our data highlight the association between iron-mediated metabolic homeostasis and the clinical efficacy and toxicity of TGT in RA therapy,offering guidance for the rational clinical use of TwHF-based therapy with dual therapeutic and toxic potential.

Single-cell transcriptome analysis uncovers underlying mechanisms of acute liver injury induced by tripterygium glycosides tablet in mice

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40%of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated hepatic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and therapeutic targets for liver protection.

Comparative Observation on the Effects of Radix Tripterygium Hypoglaucum Tablet and Tripterygium Glycosides Tablet in Treating Erosive Oral Lichen Planus

Objective: To compare the therapeutic effects of Radix Tripterygium hypoglaucum tablet (THT) and Tripterygium glycosides tablet (TGT) in treating erosive oral lichen planus(EOLP).Methods: The patients were randomized into two groups, and they were treated with THT (n=47) or TGT (n=47), respectively. The therapeutic effects were evaluated after 3 months treatment.Results: For the patients of grade 1, the total efficacy in TGT group was 85.71%, compared with 52.38% in THT group, the efficacy was statistically greater in the group receiving TGT (P=0.043). However, for the patients of grade 2, the difference was not statistically significant (P=0.173).Conclusion: TGT is more effective in treating EOLP than THT for grade 1 patients. However, TGT is not suitable for patients of child bearing age.

Characteristics and pharmacokinetics of tripterygium glycosides nano-carries transdermal delivery systems:skin-blood synchronous microdialysis and numerical simulation

The traditional Chinese medicine tripterygium glycosides(TPG)is used clinically to treat some Rheumatism,Eczema,immunosuppression and tumor,with the activities of hypnosis,antipyretic,analgesic,antiinflammatory,allergy and antitumor.However TPG has low water solubility and low skin permeability,so its clinical use is limited.Transdermal delivery systems can provide a controlled drug release rate that can keep constant concentrations of drug in the plasma for up to multiple days,improved patient compliance,and the possibility ofreducing the rate and severity of side effects.In this study,a fast and sensitive technique skin-blood two sites synchronous microdialysis coupled with LC-MS was used to study the pharmacokinetic parameter of three different formulations(TPG nanoemulsion,TPG nanoemulsion based gels and TPG gel).Creating a multilayer model,use the model to simulate the three formulations dynamics in transdermal-drug delivery system.The experiment results showed that the TPG nanoemulsion,TPG nanoemulsion based gels can significantly raise the drug concentrations in skin more than that of TPG gels.The numerical simulation results indicating that TPG gel and TPG nanoemulsion are close to practical measurements,only in the concentration increase phase the numerical simulation result has some difference with the experimental results.TPG nanoemulsion based gels have significant difference with the experimental results,both in concentration increase stage and concentration decreasing stage,but its trend was same.The study shows that the skin-blood synchronous microdialysis technique provided a new method for the pharmacokinetics study of nanocarriers transdermal delivery systems.In addition,the microdialysis technique combined with mathematical modeling provides a very good platform for the further study of transdermal delivery system.

Anti-angiogenic effect of tripterygium glycosides tablets in animal models of rheumatoid arthritis:A systematic review and meta-analysis

Objectives:To explore and summarize the beneficial effects of a traditional Chinese medicine preparation,Tripterygium glycosides tablets(TGT),in rheumatoid arthritis(RA)animal models of neovascularization,and to provide a reference for future clinical applications and research on its pharmacologic mechanism.Methods:We searched the databases PubMed,Embase,Web of Science,Chinese National Knowledge Infrastructure,VIP,Wan Fang and SinoMed(China Biomedical Document Service System)to identify studies of TGT with outcome indicators of angiogenesis-related factors that were published before April2020.Subgroup analysis and meta-regression were performed for dosage and duration of TGT.Statistical tests and subgroup analysis were conducted using RevMan 5.3,and meta-regression and sensitivity analysis were conducted using STATA/SE 15.0.Results:Fourteen studies of TGT in RA rats were included in this analysis.Treatment with TGT significantly reduces synovial microvessel density and the expression of vascular endothelial growth factor(VEGF),VEGF receptor 2,hypoxia inducible factor a,c-Fos,c-Jun,angiopoietin-1 and angiopoietin-2 compared with control groups(P<.05).Subgroup analysis did not show a significant association of the mRNA levels of VEGF in synovium,assessed using quantitative real-time PCR,with duration or dosage of TGT.Meta-regression analysis also indicated that the effects of dosage and duration were not significantly associated with differences in VEGF mRNA levels.Sensitivity analysis on VEGF m RNA levels did not fundamentally change the results.Conclusions:TGT can reduce synovial neovascularization by decreasing synovial microvessel density and expression of VEGF,VEGF receptor 2,hypoxia-inducible factor a,c-Fos,c-Jun,Ang-1 and Ang-2,thereby suppressing pannus formation and bone destruction in rat models of RA.Additional well-designed studies are required to confirm these findings.

A Synoptical Introduction of The Clinical Application of Tripterygium Glycosides

Tripterygium glycosides (TG) refers to the total glycosides, mainly the epoxy diterpene lactones extracted from the root of Tripterygium Wilfordii Hook f (TW), a common vine-like toxic plant grown in the wide area of South China. It possesses potent anti-inflammatory and immune suppressive effects and also some anti-coagulation and bacteriocidal action, and compared with other preparations of TW, it shows the superiority of being small in dose and having less adverse effect. It has been reported to be effective in treating patients with such diseases as rheumatoid arthritis, lupus or purpura nephritis, psoriasis, erythroderma, and allergoses. This article is attended to synoptically introduce the recent clinical applications of TG.

Alprostadil combined with tripterygium glycosides tablet in treatment of diabetic kidney disease: a systematic review and meta-analysis

Background:Diabetic kidney disease is now the principal cause of end-stage renal failure worldwide.Alprostadil combined with tripterygium glycosides tablet is a new method for diabetic kidney disease treatment.However,there are currently few systematic reviews on treatment for alprostadil combined with tripterygium glycosides tablet.Therefore,a systematic review and meta-analysis was conducted to analyze the function of alprostadil combined with tripterygium glycosides tablet in the treatment of diabetic kidney disease.Methods:We searched Pubmed,Embase,the Cochrane Library,Chinese databases,and clinical trial for randomized controlled trials of alprostadil combined with tripterygium glycosides tablet in the treatment of diabetic kidney disease,including results from the foundation of database until August 5,2020.Two reviewers have independently performed literature screening,data extraction,and quality evaluation.This meta-analysis has been carried out by RevMan5.4 software.Results:Ten randomized controlled trials with 724 patients were involved.Compared with alprostadil alone,the combination of alprostadil and tripterygium glycosides tablet in treatment of diabetic kidney disease could reduce the level of 24-hour urine protein(95%CI(–2.05,–0.22),P=0.01),serum creatinine(95%CI(–5.01,–0.20),P=0.03),level of interleukin-6(95%CI(−4.57,−2.37),P<0.00001),tumor necrosis factor-α(95%CI(−4.57,−2.37),P<0.00001).The combined treatment could also improve the clinical efficacy(95%CI(1.09,1.25),P<0.0001),and reduce the occurrence of serious adverse events(95%CI(0.26,0.94),P=0.03).However,there is no association of two treatments in blood urea nitrogen(95%CI(–4.17,2.11),P=0.52),albumin(95%CI(–1.10,0.97),P=0.90),triglyceride(95%CI(−1.44,1.50),P=0.97).Conclusion:Alprostadil combined with tripterygium glycosides tablet contributes to protecting renal function,inhibiting inflammation,and reducing the occurrence of adverse events,which could be considered as a feasible therapy for diabetic kidney disease patients.However,some clinical variables did not accurately conclude due to the low quality of methodology and small sample sizes.More rigorous and more extensive trials are essential to validate our results.Trial registration:Systematic review registration:(CRD42020203725).

Study on Effect of Small Dosage Tripterygium Glycosides in Auxiliary Treatment of Intractable Still's Disease

Intractable Still’s disease, namely the intractable systemic juvenile idiopathic arthritis (JIA) is a clinical difficulty of pediatrics, and so far there still lacks any special treatment. In virtue of the markedly anti-inflammatory and immunosuppres-sive effects of tripterygium glycosides (TG, product of Huangshi Pharmaceutical Factory, 10 mg/

A Dietary Supplement Jinghuosu Ameliorates Reproductive Damage Induced by Tripterygium Glycosides

Objective To determine the possible protective effects of Jinghuosu,a dietary supplement(DS),on tripterygium glycosides(TG)-induced reproductive system injury in rats and its underlying mechanisms.Methods A reproductive damage model was established in rats by feeding of TGs.Twenty-eight male Sprague Dawley rats were randomly divided into 4 groups using a random number table(n=7 in each):control(C)group,model(M)group,DS group and L-carnitine(LC)group.Rats in M,DS and LC groups received 40 mg/kg TGs orally.Starting from the 5th week,after administration of TGs for 4 h every day,rats in DS and LC groups were administered with 2.7 g/kg DS and 0.21 g/kg LC,respectively,for protective treatment over the next 4 weeks.Rats in Group C continued to receive the control treatment.Hematoxylin-eosin staining was used for histopathological analysis of rat testicular tissues.Enzyme-linked immunosorbent assay was performed to measure alkaline phosphatase(ALP),lactate dehydrogenase,alcohol dehydrogenase,total antioxidant capacity(T-AOC),superoxide dismutase,glutathione peroxidase(GSH-Px),and malondialdehyde(MDA)concentrations.Chemiluminescence assay was used to determine the serum testosterone content.Quantitative real-time PCR and Western blotting were conducted to analyze the expression of genes and proteins related to the testosterone synthesis pathway and the nuclear factor erythroid 2-related factor 2/heme oxygenase 1 antioxidant pathway.Results Oral administration of TGs induced significant increases in the testicular levels of zinc transporter 1 and MDA(P<0.05).On the other hand,sperm concentration,sperm motility,and serum testosterone,serum zinc,testicular zinc,Zrt-,Irt-like protein 1,ALP,luteinizing hormone(LH)receptor,steroidogenic acute regulatory protein,Cytochrome P450 family 11 subfamily A member 1,3β-hydroxysteroid dehydrogenase 1 T-AOC,GSH-Px,nuclear factor erythroid 2-related factor 2,heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1 levels decreased following TGs exposure(P<0.05).All of these phenotypes were evidently reversed by DS(P<0.05).Conclusion DS Jinghuosu protects against TG-induced reproductive system injury in rats,probably by improving zinc homeostasis,enhancing the testosterone synthesis and attenuating oxidative stress.

Effectiveness and safety of tripterygium glycosides tablet(雷公藤多苷片)for lupus nephritis:a systematic review and Meta-analysis

OBJECTIVE:To investigate the effectiveness and safety of tripterygium glycosides(TG)tablet(雷公藤多苷片)for the treatment of Lupus nephritis(LN).METHODS:Several databases were systematically searched including Pub Med,Embase,Cochrane,Wiley,China National Knowledge Infrastructure Database,Sino Med and Wanfang Library till June 20,2020.Revman5.3 was utilized to analyze the data according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Statement.RESULTS:In total,8 randomized controlled trials involving 583 participants were identified.Meta-analyses showed that,compared with glucocorticoids(GC)alone,the combination with TG tablet provided a statistically significant improvement in total remission(TR)(RR=1.27,95%CI:1.08–1.50,P=0.004),complete remission(CR)(RR=1.61,95%CI:1.05–2.47,P=0.03)and C3 levels(WMD=0.27,95%CI:0.14–0.39,P<0.0001),C4 levels(WMD=0.12,95%CI:0.07–0.17,P<0.00001).No significant differences were seen in TR,CR,proteinuria,serum creatinine,C3 and C4(TR:RR=1.00,95%CI:0.87–1.16,P=0.95;CR:RR=1.10,95%CI:0.78–1.56,P=0.58;proteinuria levels:WMD=-0.06,95%CI:-0.13 to 0.01,P=0.10;serum creatinine levels:WMD=-0.01,95%CI:-7.36 to 7.35,P=1.00;C3 levels:WMD=0.01,95%CI:-0.06 to 0.07,P=0.84;C4 levels:WMD=-0.01,95%CI:-0.03 to 0.01,P=0.49)between azathioprine(AZA)/leflomit(LEF)+GC and TG tablet+GC.Adverse events(hepatic dysfunction,nausea,vomitting)showed no statistical differences between the TG tablet+GC group and the GC group.There were more new onset of irregular menstruation in the TG tablet+GC group than those in the AZA+GC(RR=3.57,95%CI:1.40–9.11,P=0.008)/LEF+GC(RR=6.69,95%CI:2.42-18.46,P=0.0002)group,but leucopenia lower than those in AZA+GC group(RR=0.38,95%CI:0.17-0.85,P=0.02)and alopecia(RR=0.14,95%CI:0.03-0.77,P=0.02)and rash(RR=0.09,95%CI:0.01-0.69,P=0.02)lower than those in LEF+GC group.Conclusions:This review indicates that TG tablet maybe effective in LN treatment.Nevertheless,adverse events cannot be ignored.Large sample,multi-center,highquality clinical studies are needed to verify the exact effects and safety of TG tablet in treatment of LN.

COMPREHENSIVE TWO-DIMENSIONAL LIQUID CHROMATOGRAPHY COUPLED WITH QUADRUPLE TIME-OF-FLIGHT MASS SPECTROMETRY FOR CHEMICAL CONSTITUENTS ANALYSIS OF TRIPTERYGIUM GLYCOSIDES TABLETS

Comprehensive two-dimensional liquid chromatography platform(LC×LC)coupled with quadrupole time-of-flight(QTOF)mass spectrometry(MS)is developed to separate,identify and relatively determine the chemical constituents of two types of tripterygium glycosides tablets(TGT).The types and relative contents of the constituents discovered in two kinds of TGT tablets were subsequently compared.C8andC18 column were used for the separation of the first

Clinical Effect of Tripterygium Glycosides Combined with Glucocorticoids in the Treatment of Refractory Nephrotic Syndrome Patients: A Systematic Review and Meta-Analysis

The objective of this study is to evaluate the effectiveness and safety of Tripterygium glycosides combined with glucocorticoids for the treatment of refractory nephrotic syndrome(NS).Computer search of Chinese and English databases,including CNKI,VIP,Wan Fang Database,PubMed,Cochrane Library,Embase,and Sinomed,for randomized controlled trials(RCTs)of Tripterygium glycosides combined with glucocorticoids for refractory NS(RNS)was conducted.Meta-analysis was performed using RevMan5.3.Thirteen RCTs comprising 994 patients were included in the study.Tripterygium glycosides combined with glucocorticoids had a statistical significance on the effective rate(odds ratio[OR]=4.69,95%confidence interval[CI]3.29,6.67,P<0.00001),24-h urine protein(Weighted mean difference(MD)=-0.57,95%CI[-0.62,-0.51],P<0.00001),serum albumin(MD=4.77,95%CI[4.30,5.24],P<0.00001),total serum protein(MD=9.45,95%CI[8.73,10.17],P<0.00001),urea nitrogen(MD=-0.53,95%CI[-0.90,-0.17],P=0.005),and serum creatinine(MD=-8.45,95%CI[-15.32,-1.57],P=0.02).There was no statistical significance on adverse reactions(OR=0.68,95%CI[0.41,1.12],P=0.13).Tripterygium glycosides combined with glucocorticoids could improve clinical effective rate,reduce 24-h urine protein,improve serum albumin and total serum protein,and reduce urea nitrogen and serum creatinine levels in patients with RNS.However,the quality of the included literature is poor,and conclusion still needs further verification using larger samples and high-quality randomized,double-blind controlled trials.

CLINICAL STUDY ON EFFECT OF TRIPTERYGIUM WILFORDII HOOK. F. GLYCOSIDES ON UTERINE LEIOMYOMA

Objective To observe the therapeutic effect and side reactions of Tripterygium wilfordii Hook,f. ( GTW) glycosides on patients with uterine leiomyomas. Methods 65 normally cycling women with symptomatic uterine leiomyomas received 40mg daily dose GTW for 3 to 6 months. Baseline ultrasound tests were obtained to evaluate the sizes of myomas and uterus, then repeated three and six months after treatment. Blood samples were collected to determine the hormonal levels of in the mid-follicular and mid-luteal phases of the menstrual cycles before GTW therapy and at 3~4 months and 5~6 months after treatment. Results Significant decrease in leiomy-oma volume was shown in 39 of 65 (60% ) and 28 of 40 ( 70% ) patients after 3~4 months and 5~6 months of treatment, respectively. The decrease of the volume of leiomyoma was time-dependent as while 27. 84% and 51.60% in 3~4 months and 5 ~ 6months, respectively. 25 of 65 patients had amenorrhea during the course of treatment. Tripterygium wilfordii glycosides treatment induced a significant increase in LH and FSH levels (P < 0. 01) as compared with pretreatment values. In contrary, a significant decrease in E2 and P levels (P <0. 05) was found, but no changes were observed in T and PRL levels after treatment. Conclusion Tripterygium wilfordii might serve as an effective therapeutic agent for leiomyomas with fewer side effects. A reversible inhibitory effect on the ovary by Tripterygium wilfordii glycosides may be one of the mechanisms of Tripterygium wilfordii in decreasing leiomyoma volume.

Liangxue Xiaoban decoction and its disassembled prescriptions ameliorate psoriasis-like skin lesions induced by imiquimod in mice via T cell regulation

Objective:To explore the therapeutic capacity of the Liangxue Xiaoban(LXXB)decoction and its disassembled prescriptions in the modulation of T cell subsets and recurrence-related indexes of psoriasis using a psoriasis-like mouse model.Methods:The psoriasis model was generated by the treatment of BALB/c mice(n=48)with imiquimod.Mice were divided into six groups:control,psoriasis model,tripterygium glycosides,LXXB decoction,Liangxue decoction,and Qingqi decoction.After the intervention period,the interleukin(IL)-17A,IL-22,and interferon-γ levels in mice were examined and hematoxylin and eosin staining was conducted to determine pathological changes in the skin tissues.T cell subset changes in the skin-draining lymph nodes were analyzed using flow cytometry,and the expression levels of the associated transcription factors and recurrence-related indexes in the skin tissues were determined using a polymerase chain reaction.Results:LXXB decoction attenuated the levels of CD8^(+)T,Th17,and Th1 cells and induced an increase in the Th2 and Treg cell levels.The disassembled prescriptions promoted or inhibited specific subsets of T cells to improve the symptoms of psoriasis.Notably,the LXXB and Liangxue decoctions suppressed the expression of IL-22 at both the gene and protein levels and restored the CD103 and IL-15 expressions in the skin tissue to the normal range.Conclusion:LXXB decoction exerted significant immunoregulatory effects on T cell subsets and improved the recurrence-related indexes.Interestingly,the Liangxue prescription appeared to have a therapeutic advantage in terms of Th17 modulation and psoriasis recurrence,while the Qingqi prescription performed better in Treg immunoregulation.

Integrating systematic pharmacology-based strategy and experimental validation to explore mechanism of Tripterygium glycoside on cholangiocyte-related liver injury

Objective:Tripterygium glycoside(TG)is widely used in clinical practice for its multiple bioactivities including anti-inflammatory and immunosuppressive effects.However,emerging studies have frequently reported TG-induced adverse reactions to multiple organs,especially liver.Here,this study aimed to investigate the mechanism of liver damage induced by TG and explore representative components to reflect TG hepatotoxicity.Methods:Network pharmacology was used to determine the potential targets of bile duct injury caused by TG.Next,the hepatotoxic effects of TG,triptolide(TP)and celastrol(CEL)were investigated and compared in vivo and in vitro.Liver function was determined by measuring serum transaminase and histopathology staining.The cell proliferation and apoptosis were determined by cell viability assay,scratch assay and flow cytometry.The expression of gene of interest was determined by qPCR and Western blot.Results:Based on the network pharmacological analysis of 12 bioactive ingredients found in TG,a total of35 targets and 15 pathways related to bile duct injury were obtained.Both TG and TP resulted in cholangiocyte damage and liver injury,as illustrated by increased levels of serum transaminase and oxidative stress,stimulated portal edema and lymphocytic infiltration and decreased expression of cholangiocyte marker,cytoskeletal 19.In addition,TG and TP inhibited cell proliferation and migration,arrested cell cycle and promoted Caspase-dependent apoptosis of cholangiocytes via suppressing the phosphorylation of extracellular regulated protein kinases 1/2(ERK1/2)and protein kinase B(AKT).While,CEL at equivalent dosage had no obvious hepatotoxicity.Conclusion:We revealed that TG-stimulated liver injury was specifically characterized by cholangiocyte damage and TP might be the decisive ingredient to reflect TG hepatotoxicity.Our results not only provide novel insights into the mechanism underlying the hepatotoxicity effects of TG but also offer reference for clinical rational use of TG.

Treatment of Fibrillary Glomerulonephritis by Corticosteroids and Tripterygium Glycoside Tablets:A Case Report

Fibrillary glomerulonephritis （FGN） is a rare kidney disease characterized by the deposition of randomly arranged fibril deposits 15-30 nm in diameter, mainly in the mesangium and/or glomerular basement membranes （GBMs）, and negative Congo red staining.（1＇2） The diagnosis of FGN is made in about 1% of native kidney biopsy specimens.（3）