Induced Intermediate Mesoderm Combined with Decellularized Kidney Scaffolds for Functional Engineering Kidney

BACKGROUND: Chronic kidney disease is a severe threat to human health with no ideal treatment strategy. Mature mammalian kidneys have a fixed number of nephrons, and regeneration is difficult once they are damaged. For this reason, developing an efficient approach to achieve kidney regeneration is necessary. The technology of the combination of decellularized kidney scaffolds with stem cells has emerged as a new strategy; however, in previous studies, the differentiation of stem cells in decellularized scaffolds was insufficient for functional kidney regeneration, and many problems remain. METHODS: We used 0.5% sodium dodecyl sulfate (SDS) to produce rat kidney decellularized scaffolds, and induce adipose-derived stem cells (ADSCs) into intermediate mesoderm by adding Wnt agonist CHIR99021 and FGF9 in vitro. The characteristics of decellularized scaffolds and intermediate mesoderm induced from adipose–derived stem cells were identified. The scaffolds were recellularized with ADSCs and intermediate mesoderm cells through the renal artery and ureter. After cocultured for 10 days, cells adhesion and differentiation was evaluated. RESULTS: Intermediate mesoderm cells were successfully induced from ADSCs and identified by immunofluorescence and Western blotting assays (OSR1 + , PAX2 +). Immunofluorescence showed that intermediate mesoderm cells differentiated into tubular-like (E-CAD + , GATA3 +) and podocyte-like (WT1 +) cells with higher differentiation efficiency than ADSCs in the decellularized scaffolds. Comparatively, this phenomenon was not observed in induced intermediate mesoderm cells cultured in vitro. CONCLUSION: In this study, we demonstrated that intermediate mesoderm cells could be induced from ADSCs and that they could differentiate well after cocultured with decellularized scaffolds.

A randomized prospective study of early steroid withdrawal in middle aged and elderly renal transplant patients / 中华老年医学杂志

Objective To evaluate the safety and validity of an early steroid withdrawal protocol including cyclosporine (CsA) and mycophenolate mofetil (MMF) in middle aged and elderly renal transplant patients. Methods Between September 2000 and April 2008, the prospective, randomized study design was used in 80 middle aged and elderly renal transplant patients. Steroid withdrawal group (n=39) with primary cadaveric kidney transplants received a protocol consisting of CsA 4～6 mg·kg~(-1)·d~(-1) beginning at postoperative day 3, MMF 0. 75 g twice a day from the next postoperative day, and methylprednisolone (MP) 500 mg daily from day 0 to 3. Then prednisone (Pred) 20 mg daily was gradually tapered and withdrawn after postoperative day 30. Conventional steroid treatment group (control group, n=41) received a regimen consisting of CsA, MMF and MP, and Pred 20 mg daily. Pred was tapered to 5 mg daily over a period of 6 months, then maintained thereafter. Outcome parameters were patient and graft survival rates, renal function, acute rejection ( AR), arterial hypertension, hyperlipidemia or diabetes mellitus, weight gain and infection. Results The incidence of AR in the steroid withdrawal group was similar to the control group (23. 1% vs. 19. 5%, χ~2=0. 15,P>0. 05). Patient survival rates at 12, 24, 36 months were 97. 4%, 94. 8% and 88.0% in the steroid withdrawal group and were 97.6%, 97.6 and 87.8% in the control group, respectively (χ~2=0. 17, P>0. 05). And graft survival rates were 94. 9%, 88. 6% and 83. 7% in the steroid withdrawal group and were 95. 1%, 91. 5% and 79. 5% in control group, respectively (χ~2 = 0.07, P>0. 05). Conclusions In middle aged and elderly renal transplant patients, early steroid withdrawal is feasible and may not significantly increase the risk of acute rejection episodes.

Expression of M3 subtype of muscarinic receptors in normal,BPH and prostate cncer tissue / 中华泌尿外科杂志

Objective To study the relationship between M3 receptor and prostastic tumor by analyzing the expressions of M3 and vascular endothelial growth factor(VEGF)in adult human normal and neoplastic prostatic gland tissue. Methods The specimens included 36 normal prostates(fresh),36 benign prostatic hyperplasia(BPH)tissue(fresh),and 36 cancer tissue(8 fresh).RT-PCR was used to detect M3 receptor,VEGPs genetic expression.At protein level,VEGF,Ms receptor,CD34 were detected by western-blot and immunohistochemical method. Results VEGF and M3 receptor's genetic expressions were higher in prostate cancer tissue(O.8354±0.1897,0.7824±0.2047)than in BPH tissue(0.6735±0.1603,0.6021±0.1637),while the expressions of these genes were lowest in normal prostate tissue(0.5425±0.1629,0.3436±0.1581)(P<0.001).There was a positive correlation between M3 and VEGFs gene expression(r=0.4999,P<O.001).The protein expression of Ms receptor and VEGF was similar to their genetic expression pattern,the expressions in prostate cancer tissue(0.4777±0.1638,0.5718±0.2245)were higher than those in BPH tissue(0.3655±0.1474,0.4342±0.1538)and in normal prostate tissue(0.2659±0.1076,0.3380±0.1527)(P<0.05).Conclusions M3 receptor might be related with development of prostatic tumor.It could be used as a new target 0f diagnosis and treatment for prostastic tumor.