Progress on electron microscopy diagnosis on Banff classification for renal allograft pathology / 器官移植

With the development of organ transplantation in clinical practice, allograft pathology has been constantly developing and advancing. The convening of Banff conference on allograft pathology and the establishment of Banff classification on allograft pathology (Banff classification) are pivotal milestones in the development of international allograft pathology. Since then, Banff classification on pathological diagnosis of various transplant organs have been continually updated and improved. Ultrastructural pathological observation by electron microscope plays an irreplaceable role in the early diagnosis of antibody-mediated rejection, recurrent disease and de novo disease of renal allograft. Early detection and rational treatment help to maintain the long-term survival of renal allograft and reduce the failure of renal allograft. In this article, the basic definition of electron microscope and the ultrastructural pathological diagnosis, the research history and main progress on electron microscope diagnosis on Banff classification for renal allograft pathology were introduced, and typical pathological changes, specific terminology and diagnostic criteria of electron microscope diagnosis on renal allograft biopsy were summarized, aiming to provide reference for clinical and basic research of organ transplantation.

Transcription factor SP1 decoy ODNs targeting α1, 3-GT renders porcine endothelial cells resistant to complement-mediated cytotoxicity / 中华器官移植杂志

Objective To investigate whether porcine endothelial cells transfected with SP1 de-coy ODNs could resist complement-mediated cytotoxicity during the model of SV-40-PED with human serum in vitro. Methods Immortalized porcine aortic endothelial cells of the line PED were divided in-to three groups. The transfected group was SV-40-PED with SP1 decoy ODNs. The mismatched group was SV-40-PED with scrambled SP1 decoy ODNs. The negative group was cells with oligo-fectamine only. The expression of α1,3-GT mRNA and αGal was detected after 26 h by using fluores-cence microscope, Western blot, RT-PCR and lactate dehydrogenase (LDH) activity assay. Results Fluorescence microscopy observed the decreased fluorescence of αGal after SP1 decoy ODNs transfec-tion. Dotted fluorescent decrease could be observed on some membrane while the mismatched group and negative group with bright green fluorescence. Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was decreased to 52.6% of the negative group (P<0.05). The expression of α1,3-GT mRNA in the PED cells transfected with decoy ODNs was 0.42±0.20 (isoform 1) and 0.27±0.12 (isoform 2) respectively, significantly lower than in the negative group (isoform 1, 0.72±0.17; isoform 2, 0.50±0.19; both P<0.05). The expression of α1, 3-G Tin the mismatch group was not different from that in the negative group (P>0.05). 20% normal hu-man serum (NHS) and 40 % NHS had cytotoxic effect on both mismatch and negative groups, but de-coy ODNs could confer SV-40-PED protection from the cytolysis effect (P<0.05), which made a re-markable reduction of complement-mediated cytotoxicity towards SV-40--PED. Conclusions SP1 decoy ODNs could confer porcine endothelial cells protection from complement-mediated cytotoxicity effect in vitro.

IL-5 Up-regulates the Expression of TGF-β1 in Human Blood Eosinophils in Vitro / 华中科技大学学报（医学）（英德文版）

To investigate the effects of IL-5 on the expression of TGF-β1 in eosinophils in vitro, eosinophils were incubated in the presence of the same concentrations of IL-4, IL-5 and IFNγ, different concentrations of IL-5 in vitro and changes of eosinophil viability were assessed by trypan blue exclusion. Non-cytokine was employed as a negative control. 16 h after the cultivation, supernatants and cells were assayed by using TGF-β1 specific ELISA and RT-PCR. The mRNA expression and protein expresssion of TGF-β1 in eosinophils stimulated with different cytokines was observed.The expression of TGF-β1 protein in eosinophils was increased significantly by IL-4 (433.67±9.86vs 228.9±2.87) and IL-5 (403. 72±7.60 vs 228.9±2.87, P＜0.05), while decreased by IFNγ (178.47±2.60 vs 228.9±2.87). At the same time, the results demonstrated that the basal level of TGF expression was enhanced by IL5 in all samples (P＜0.05). The expression of TGF β1 mRNA was 1.42, 1. 70, 1. 76-folds higher than that of the non-stimulated controls. It is concluded that IL-5 can up-regulate the expression of TGF-β1 in eosinophils in vitro, which might have effect in eosinophil-associated chronic rejection.

The effect of transcription factor SP1 decoy oligodeoxynucleotides on expression of ? Gal in SV-40-PED cells / 医学研究生学报

Objective:To investigate the role of transcription factor SP1 decoy oligodeoxynucleotides(ODN) on expression of ? Gal in SV-40-PED cells.Methods:Immortalized porcine aortic endothelial cells of the PED line were cultured and transfected with ?1,3galactosyltransferase(?1,3GT) specific decoy ODN.Cells transfected with mismatch ODN was used as negative controls.Twenty-six hours later the cells were collected.The expression of ? Gal was determined with fluorescence microscope and Western blot.The expression of ?1,3GT mRNA was examined by RT-PCR.Results:Fluorescence microscopy observed the decreased fluorescence of ? Gal after decoy ODN transfection.Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was(48.2?0.9).It is 52.6% of the mock group(P0.05).Conclusion:?1,3GT gene reduce actually occurs following transfection of decoy ODN.Porcine endothelial cells can be the targets of decoy ODN.