TAC reduced Glomerular IgG and C3 deposition in MRL/lpr mice.

<p>(A) TAC administration decreases IgG deposition in glomeruli of MRL/lpr mice (starting at 12 weeks of age, 6 per group), as shown by immunoflourescence. (B) The mean fluorescence intensity corresponding to IgG staining/deposition (n = 6). (C) TAC treatment decreases C3 deposition in glomeruli. (D) The mean fluorescence intensity corresponding to C3 staining/deposition (n = 6). Magnification, ×400 in A and C. Nuclei are counterstained with DAPI. NC, normal control group; DC, disease control group; Tx, treatment group. * <i>P</i> < 0.05.</p

TAC preserved podocyte number and reduced podocyte apoptosis.

<p>(A) WT1 expression in MRL/lpr mice (starting at 12 weeks of age, 6 per group), as shown by immunoflourescence. (B) Podocyte number per mean glomerular tuft in each group (n = 6). (C) Real time-PCR analysis of WT1-mRNA expression was performed, n = 4. (D, E) Mouse podocyte cells were pre-incubated with TAC (10 μM or 20 μM) followed by treatment with TGF-β₁ (5 ng/ml or 10 ng/ml); n = 4. Apoptosis was assessed with annexin V by flow cytometery (FCM). TGF5, TGF-β₁ 5 ng/ml; TGF10, TGF-β₁ 10 ng/ml; TAC10, TAC 10 μM; TAC20, TAC 20 μM. NC, normal control group; DC, disease control group; Tx, treatment group. * <i>P</i> < 0.05, ^#<i>P</i> < 0.01.</p

TAC restored synaptopodin expression and stabilized podocyte cytoskeleton.

<p>(A) TAC restored synaptopodin expression in MRL/lpr mice (starting at 12 weeks of age, 6 per group), as shown by immunofluorescence. (B) Western blot showing synaptopodin expression in each group (n = 6). (C) Real time-PCR showing synaptopodin mRNA in each group (n = 6). (D) Treatment with TAC attenuated the decrease in F-actin staining (phalloidin staining) induced by TGF-β1 (5ng/ml) in mouse podocytes (MPC5 cells). NC, normal control group; DC, disease control group; Tx, treatment group. * <i>P</i> < 0.05, ^#<i>P</i> < 0.01.</p

TAC attenuated renal pathology damage in MRL/lpr mice.

<p>(A) At the end of the treatment period (8 weeks), kidney tissues from mice (starting at 12 weeks of age) were immersion-fixed in 4% paraformaldehyde/phosphate buffered saline and embedded in paraffin. Sections (2 μm) were stained with HE, PAS and Masson stain. There were 8 and 9 mice in DC 8w and Tx 8w group, respectively; 6 per group in other groups. (B) Thirty glomeruli for each kidney section were digitally quantified. A microscope equipped with a color camera was used and data were analyzed using image analysis software. According to the glomerular, renal tubular and pathology rating criteria [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132724#pone.0132724.ref005" target="_blank">5</a>], we calculated pathological scores in each mouse. NC, normal control group; DC, disease control group; Tx, treatment group. DC 8w vs. NC 8w, <i>P</i> < 0.01; DC 8w vs. Tx 8w, <i>P <</i> 0.05.</p

TAC prevented the effacement of foot process in MRL/lpr mice.

<p>(A) TAC prevented the effacement of foot process (arrows) and preserved the slit diaphragm in MRL/lpr mice (starting at 12 weeks of age, 6 per group), as shown by transmission electronic microscopy (13,500×). (B) The number of slit pores per 100 μm length of glomerular basement membrane (GBM). FP, foot process. NC, normal control group; DC, disease control group; Tx, treatment group. * <i>P</i> < 0.01.</p

TAC reduced proteinuria and preserved renal function in MRL/lpr mice.

<p>(A) All animals (starting at 12 weeks of age, 6 per group) were placed in metabolic cages for urine collection before therapy (0 day) and at the end of the study (8 weeks treatment). Urine samples were used to examine the levels of 24 hour urinary protein excretion. (B, C) Blood samples were obtained from MRL/lpr mice (starting at 12 weeks of age, 6 per group) by eye puncture under ether anesthesia to examine the levels of BUN and serum creatinine just before all mice were killed at 0 week and 8 weeks. NC, normal control group; DC, disease control group; Tx, treatment group. *<i>P</i> < 0.05.</p

Value of original and modified pathological scoring systems for prognostic prediction in paraffin-embedded donor kidney core biopsy

No study has validated, compared and adapted scoring systems for prognosis prediction based on donor kidney core biopsy (CB), with less glomeruli than wedge biopsy. A total of 185 donor kidney CB specimens were reviewed using seven scoring systems. The association between the total score, item scores, score-based grading, and allograft prognosis was investigated. In specimens with less than ten glomeruli (88/185, 47.6%), scoring systems were modified by adjusting weights of the item scores. The Maryland aggregate pathology index (MAPI) score-based grading and periglomerular fibrosis (PGF) associated with delayed graft function (DGF) (Grade: OR = 1.59, p p = 0.006). Total score, score-based grading and chronic lesion score in scoring systems associated with one-year and 3-year eGFR after transplantation. Total-score-based models had similar predictive capacities for eGFR in all scoring systems, except MAPI and Ugarte. Score of glomerulosclerosis (GS), interstitial fibrosis (IF), tubular atrophy (TA), and arteriolar hyalinosis (AH) had good eGFR predictive capacities. In specimens with less than ten glomeruli, modified scoring systems had better eGFR predictive capacities than original scoring systems. Scoring systems could predict allograft prognosis in paraffin-embedded CB with ten more glomeruli. A simple and pragmatic scoring system should include GS, IF, TA and AH, with weights assigned based on predictive capacity for prognosis. Replacing GS scores with tubulointerstitial scores could significantly improve the predictive capacity of eGFR. The conclusion should be further validated in frozen section.</p