Improvement of human embryonic stem cell-derived retinal pigment epithelium cell adhesion, maturation, and function through coating with truncated recombinant human vitronectin

AIM:To explore an xeno-free and defined coating substrate suitable for the culture of H9 human embryonic stem cell-derived retinal pigment epithelial(hES-RPE)cells in vitro,and compare the behaviors and functions of h ESRPE cells on two culture substrates,laminin521(LN-521)and truncated recombinant human vitronectin(VTN-N).METHODS:hES-RPE cells were used in the experiment.The abilities of LN-521 and VTN-N at different concentrations to adhere to hES-RPE cells were compared with a high-content imaging system.Quantitative real-time polymerase chain reaction was used to evaluate RPE-specific gene expression levels midway(day 10)and at the end(day 20)of the time course.Cell polarity was observed by immunofluorescent staining for apical and basal markers of the RPE.The phagocytic ability of hES-RPE cells was identified by flow cytometry and immunofluorescence.RESULTS:The cell adhesion assay showed that the ability of LN-521 to adhere to hES-RPE cells was dosedependent.With increasing coating concentration,an increasing number of cells attached to the surface of LN-521-coated wells.In contrast,VTN-N presented a strong adhesive ability even at a low concentration.The optimal concentration of LN-521 and VTN-N required to coat and adhesion to hES-RPE cells were 2 and 0.25μg/cm^(2),respectively.Furthermore,both LN-521 and VTN-N could facilitate adoption of the desired cobblestone cellular morphology with tight junction and showed polarity by the hES-RPE cells.However,hES-RPE cells cultivated in VTN-N had a greater phagocytic ability,and it took less time for these hES-RPE cells to mature.CONCLUSION:VTN-N is a more suitable coating substrate for cultivating hES-RPE cells.

IgG4-related kidney disease complicated with retroperitoneal fibrosis:A case report

BACKGROUND IgG4-related disease(IgG4-RD)is a chronic fibrotic disease mediated by immunity recognized by clinicians in recent years.When the kidney is involved,it is called IgG4-related kidney disease(IgG4-RKD).IgG4-related tubulointerstitial nephritis(IgG4-TIN)is a representative manifestation of IgG4-RKD.IgG4-TIN can cause obstructive nephropathy complicated by retroperitoneal fibrosis(RPF).Cases of IgG4-TIN complicated with RPF are rare.Glucocorticoids are the firstline therapeutic medication for IgG4-RD and can significantly improve renal function.CASE SUMMARY Herein,we report the case of a 56-year-old man with IgG4-RKD complicated with RPF.The patient presented to the hospital with complaints of elevated serum creatinine(Cr),nausea,and vomiting.During hospitalization,Cr was 1448.6μmol/L,and serum IgG4 was increased.A total abdominal computed tomography(CT)scan and enhanced CT scan obviously indicated RPF.Although this patient had a long course and renal insufficiency,we performed a kidney biopsy.Renal biopsy showed that the renal tubulointerstitium had focal plasma cell infiltration and increased lymphocyte infiltration accompanied by fibrosis.After combining the biopsy results with immunohistochemistry,it was found that the absolute number of positive IgG4+cells per high power field exceeded 10,and the ratio of IgG4/IgG was over 40%.Finally,the patient was diagnosed with IgG4-TIN complicated with RPF and given glucocorticoids as long-term maintenance therapy,helping him keep out of dialysis.After a follow-up of 19 mo,the patient had recovered well.Previous literature on IgG4-RKD and RPF was retrieved from PubMed to characterize the clinical and pathological features and to identify the diagnosis and treatment of IgG4-RKD.CONCLUSION Our case report demonstrates the clinical characteristics of IgG4-RKD complicated with RPF.Serum IgG4 is a favorable indicator for screening.Performing renal biopsy actively plays a vital role in diagnosis and treatment,even if the patient has a long course and manifests with renal insufficiency.It is remarkable to treat IgG4-RKD with glucocorticoids.Hence,early diagnosis and targeted therapy are essential for reversing renal function and improving extrarenal manifestations in patients with IgG4-RKD.

经食管三维超声在二尖瓣成形术中的应用

目的探讨经食管实时三维超声心动图(RT3D-TEE)在二尖瓣成形术中的应用价值。方法 18例拟行二尖瓣成形术的二尖瓣脱垂患者,术前及术后即刻均接受RT3D-TEE检查。术前观察二尖瓣脱垂的三维特征及小叶区域,与外科术中所见对照;术后即刻评估二尖瓣成形的手术效果,比较手术前后二尖瓣的对合高度、瓣环周长和反流面积。结果 RT3D-TEE显示二尖瓣脱垂的三维图像与外科视野所见高度一致,术前诊断脱垂小叶的敏感性、特异性分别为96.00%、100.00%。18例患者中有2例转行二尖瓣置换术,其余16例成功完成二尖瓣成形术。RT3D-TEE观察术后二尖瓣叶及人工瓣环的形态良好,二尖瓣反流程度明显改善。二尖瓣的对合高度较术前增加,瓣环周长、反流面积较术前减少,差异有统计学意义(t=-13.556,37.789和20.847,均P=0.000)。结论 RT3D-TEE能清晰显示二尖瓣的三维形态,术前准确诊断脱垂小叶及术后即刻评估手术效果,在二尖瓣成形术中发挥重要的作用。