1.The Yi-Qi-Bu-Shen Recipe Attenuates High Glucose-induced Podocyte Injury Via The Inhibition Of NF?B And MAPK Signaling Pathway 2.Predictive Factors For Early Hypothyroidism And Early Hepatic Dysfunction Following The Radioactive Iodine Therapy In Graves

BackgroundDiabetic nephropathy(DN)is one of the most serious complications of diabetes mellitus and the main cause of the end-stage renal disease(ESRD).Microalbuminuria and later proteinuria are the most important clinical manifestations in the developing process of DN.The glomerular filtration barrier(GFB),which prevents proteins leaking from capillaries to urine,includes endothelial cells,glomerular basement membrane(GBM)and podocytes.Any injury of these portions may lead to the occurrence of microalbuminuria and later proteinuria.Podocytes are highly differentiated epithelial cells that interface with the GBM surface and play a significant role in maintaining the integrity of the GFB.Increasing evidence suggests that podocytes prevent protein leakage from plasma to primary urine.Slit diaphragms(SD)between podocytes are the essential determinant of podocyte function.Some specific proteins in the SD could affect the structure and function of podocytes,the most important of them are nephrin,podocin and CD2AP.Nephrin is a transmembrane glycoprotein and the structural backbone of SD,which encoded by the NPSH1 gene.Podocin is a hair-like structured membrane protein expressed specifically in SD,which can enhance the signal transduction induced by nephrin and is involved in the structure of cytoskeleton.CD2AP is a cohesive protein that connects SD and the cytoskeleton and plays a stabilizing role in podocytes.In DN patients and animal models,the mRNA and protein expressions of these molecules are significantly reduced and closely related to the progression of proteinuria.Hence,finding a drug that can protect these molecules from injury is essential for DN treatment.Chronic inflammation is closely associated with permeability changes in the GFB and proteinuria in DN.Inflammatory cytokines,such as IL-1?,IL-6 and TNF-?,are elevated even in the early stage of DN.Subsequently,TNF-? amplifies the activation of inflammatory cytokines,thereby leading to DN progression.Among several signaling pathways that regulate chronic inflammation,it has been reported that NF?B and MAPK are associated with podocyte injury in DN.The Yi-Qi-Bu-Shen recipe(YB)has been used in Qi'lu Hospital for more than 20 years,previously clinical data demonstrated that YB reduces proteinuria and decreases blood glucose level in DN patients.However,the exact mechanisms of YB in the treatment of proteinuria remain unknown.Since podocyte injury has been found to be the primary determinant of proteinuria,the aim of the present study is to investigate the protective effects of YB on high glucose(HG)-induced podocyte injury.Methods1.Culture conditionally immortalized human podocytes to do the following experiments.2.Cell viability measured by LDH assay.3.Gene expression detected by Q-PCR.4.Transcriptional activity of NF?B detected by Dual-Luciferase reporter assay system.5.Protein expression detected by Western blot.6.Statistical analysis.Results1.The influence of HG and YB on cell viability in podocytes.The viability of podocytes was determined by the LDH assay.Podocytes treated with HG and mannitol(up to 40 mmol/L for 24h)not reveal an effect of HG upon cell viability.YB did not affect cell viability in either condition below the 200 ?g/mL YB dose.Thus,subsequent experiments employed a 100 ?g/mL dose of YB which was demonstrated to have no effect on cell death.2.The effects of HG and YB on nephrin,podocin and CD2AP expression.HG remarkably decreased the expression of nephrin,podocin and CD2AP at both mRNA and protein levels.Interestingly,co-treatment with YB attenuated the effect of HG evidenced through significantly restored mRNA and protein expressions of the three markers,suggesting a therapeutic action of YB in HG-induce podocyte injury.3.YB suppresses the pro-inflammatory cytokines in podocytes.RT-PCR revealed that the mRNA expressions of IL-1?,IL-6,TNF-? and MCP-1 were all up-regulated in the HG group compared with the control group.Treatment with YB could significantly inhibit the transcriptional activity of these inflammatory cytokines induced by HG.These results revealed a potent anti-inflammatory profile of YB in podocytes.4.Effect of YB on NF?B signaling pathway.HG increased transcriptional activity of NF?B compared with the control group.Moreover,the phosphorylation of IKK?/? and I?B? were also up-regulated in the HG group,suggesting activation of NF?B signaling pathway.However,the protein levels of p-IKK?/? and p-I?B? were decreased after YB treatment.Furthermore,the transcriptional level of NF?B was also down-regulated by YB,which demonstrates the inhibitory effects of YB on NF?B signaling pathway.5.Effect of YB on MAPK signaling pathway.HG significantly activated the phosphorylation of ERK and P38,but only slightly increased the protein level of p-JNK.After co-treatment of YB,the phosphorylation level of ERK and P38 can be significantly reduced,suggesting that YB may exert its anti-inflammatory effect by inhibiting the ERK and P38 signaling pathway.6.Effect of inhibitors on nephrin,podocin and CD2AP expression.The I?B? inhibitor(BAY 11-7085),ERK inhibitor(U0126)and P38 inhibitor(SB203580)could partly reverse the reduced expression of nephrin,podocin and CD2AP observed in HG conditions,which further suggests that YB attenuated the podocyte injury induced by HG through the NF?B and MAPK signaling pathways.Conclusions1.YB could attenuate the podocyte injury induced by HG,the specific points are nephrin?podocin and CD2AP.2.YB attenuate the podocyte injury induced by HG through the NF?B and MAPK signaling pathways.BackgroundHyperthyroidism is a clinical syndrome caused by increased thyroid hormone in blood,it can lead to multiple complications,including cardiac,hepatic and hematologic system complications.More than 80% of the hyperthyroidism are caused by Graves5 disease(GD),and 3% of women and 0.5% of men may suffer GD in their lifetime.The incidence is 20 to 30 cases per year per 100,000 persons.The incidence peaks between 30 and 60 years of age,but people can be affected at any age.Common symptoms are palpitations,fatigue,tremor,anxiety,disturbed sleep,weight loss9 heat intolerance,sweating,and polydipsia.GD can be treated by anti-thyroid drugs(ATD),radioactive iodine(RAI)or thyroidectomy.ATD have lower risk of permanent hypothyroidism compared to RAI or thyroidectomy,but they are likely to cause frequently mild side effects and some rare but severe side effects,such as rash,vasculitis,agranulocytosis and acute hepatonecrosis,so the patients taking medications requires frequent hematological examinations.Commonly,after 12-18 months5 drag treatment,the risk of recurrence is around 50%.Compared to ATD? RAI is a definitive treatment of hyperthyroidism and can improve hyperthyroidism quickly.Due to its low cost and high efficiency,lots of patients are tend to use RAI therapy to cure GD all over the world.Although the side effects of RAI are rare,mild and transient,it could result in hypothyroidism frequently.The aim of this study is to examine the occur of early hypothyroidism in GD patients treated with RAI in our center between January 2017 to December 2018,and to identify the significant risk factors influencing the occurrence of early hypothyroidism.MethodsWe reviewed 312 GD patients treated with RAI between January 2017 to December 2018,collected the potential risk factors,and analyzed the relationship between these variables and early hypothyroidism.The main methods including:1.Patients inclusion2.Radioactive iodine uptake measurements3.Thyroid volume and weight estimation4.Radioactive iodine Dose Calculation5.Radioactive iodine therapy6.Follow-up7.Statistical analysisResults1.After 6 months9 follow-up,218(69.87%)patients were evaluated as early hypothyroid.2.Male gender,shorter duration of disease,smaller thyroid weight,lower 2-hour radioactive iodine uptake(RAIU),6-hour RAIU? 24-hour RAIU and 6/24-hour uptake ratio,lower administered dosages were significantly associated with early hypothyroidism.3.Logistics regression analysis showed that male gender,smaller thyroid weight and lower 6-hour RAIU were associated with early hypothyroidism.4.Multi-factors combined ROC curve analysis suggested that the predictive power of male gender,smaller thyroid weight and lower 6-hour RAIU for early hypothyroidism was 0.711.Conclusions1.Our results show that RAI is an effective therapy for GD and most of the cured patients became to hypothyroid within 6 months.2.Male gender,smaller thyroid weight and lower 6-hour RAIU are the main risk factors for early hypothyroidism.BackgroundGraves' disease(GD)is the most important cause of hyperthyroidism,the number of incidences are between 2.6 to 120 per 100,000 per year among different countries,the incidence peaks from 30 to 60 years of age and the lifetime risk is 3%for women and 0.5% for men.The main symptoms of GD including heat intolerance,tremor,weight loss,fatigue,anxiety,insomnia,thirst,polyuria,increased stool frequency,muscle weakness,palpitations,eye symptoms(swelling,pain,redness,double vision)and menstrual disturbances in women,which greatly affects tiie life quality of the patients.The main treatment of GD including anti-thyroid drugs(ATD),radioactive iodine(RAI)and surgical thyroidectomy.ATD,which mainly including propylthiouracil(PTU)and methimazole(MMI),remain the first-line treatment in Asia and Europe,typically take 1.5 to 2 years,and about half of the patients will relapse after withdrawal.Some recent studies showed that if the drug duration extended,the recurrence rate after withdrawal can be reduced than conventional treatment.The main side effects of ATD including hepatotoxicity,agranulocytosis,rash and anti-neutrophil cytoplasmic antibody-associated vasculitis,etc.These side effects greatly affect the clinical applications of ATD.Many patients,especially young patients,are reluctant to do the laboratory tests of leukocyte and hepatic function regularly.Due to the high cure rate and short treatment duration,many GD patients are prefer to choose RAI therapy.As the number of patients receiving RAI therapy increases,people pay more attention to the side effects of RAI therapy.As far as we know,the side effects of RAI tiierapy mainly including transient exacerbation of thyrotoxicosis,development and progression of ophthalmopathy,radioactive thyroiditis,local lymphadenitis,uncertain risks of cancer and mutagenesis.Previous studies revealed that after taking RAI,the distribution of iodine in the liver is higher than any other main organs except the thyroid.However,studies about the influence of hepatic function after RAI therapy among GD patients are still very rare.Because most of the RAI is eliminated from the body in urine,saliva,and feces in 2 days,so in this study,we performed the hepatic function tests before and within48 hours after RAI therapy among GD patients,in order to investigate the early effect of RAI therapy on hepatic function.MethodsWe reviewed 518 GD patients treated with RAI between January 2019 to July2020 in our department,collected the demographic data,results of thyroid and hepatic function tests,and analyzed the relationship between hepatic function and RAI therapy.1.Patients inclusion2.Radioactive iodine uptake measurements3.Thyroid volume and weight estimation4.Radioactive iodine Dose Calculation5.Radioactive iodine therapy6.Hepatic function tests7.Thyroid function tests8.Statistical analysisResults1.Of the 483 patients who had the full result of hepatic function tests,361patients(74,74%)had hepatic dysfunction before RAI therapy,so only 122 patients were included in the study ultimately.2.After the RAI therapy,the results of total protein(TP),albumin(ALB)5globulin(GLO),total bilirubin(TBIL),direct bilirubin(DBIL)S indirect bilirubin(IBIL),aspartate aminotransferase(AST)and gamma glutamyl transpeptidase(GGT)were statistically different from those before RAI therapy.3.26.2% of the patients developed new hepatic dysfunction,including ALB,DBIL,AST,ALT,alkaline phosphatase(ALP)and two or more abnormalities.4.Age and dose of RAI were significantly associated with hepatic dysfunction after RAI therapy.5.The predictive power of age and dose of RAI for hepatic dysfunction was0.690.6.However,none of the patients developed severe hepatic dysfunction(test results were 3 times higher than the upper limit of normal range)after RAI therapy.Conclusions1.Our results show that RAI therapy has mild early effect on hepatic function.2.Age and dose of RAI are the main risk factors for hepatic dysfunction.