Background: The optimal surgical approach for managing multiple renal arteries in living donor kidney transplantation remains unclear. This study compared outcomes between separate anastomoses (SA) and pantaloon anastomosis (PA). Methods: A retrospective cohort study was conducted from November 2018 to August 2024. In total, 117 patients were included (SA, 67; PA, 50). The primary outcome was arterial complications. Secondary outcomes included renal function, graft and patient survival at 1 year, and intraoperative variables. Statistical analyses utilized the t-test, the Mann-Whitney U-test, and Kaplan-Meier survival curves. Results: The mean warm ischemia time was similar between groups (105.48±35.64 vs.103.92±33.51 min; P=0.811). Cold ischemia time was also comparable (75.90±14.36 vs.80.68±17.85 min; P=0.111). However, anastomosis time was significantly longer in the SA group (39.00 [35.00–43.00] min) compared with the PA group (34.00 [28.50–39.50] min; P<0.001). The immediate postoperative creatinine reduction was slightly greater in the SA group (46.83%±20.68% vs. 39.78%±19.04%; P=0.090). Vascular complications requiring re-exploration were infrequent in both groups (1.5% vs. 2.0%). At 1 year posttransplant, creatinine levels were comparable between the SA (1.20 [0.95–1.45] mg/dL) and PA (1.20 [1.05–1.35] mg/dL) groups (P=0.337). Regression analysis confirmed that postoperative creatinine levels were independently associated with SA (P=0.006). Graft survival was similarly high (97% vs. 98%), as was patient survival (100% vs. 96%). Conclusions Both anastomosis techniques are safe and effective for transplanting kidneys with anatomical anomalies. SA offers surgical efficiency and provides superior early renal recovery.

Objective: Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and Methods: CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 μL/g). Results: Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.

Mixed lineage leukemia proteins (MLL) are the key histone lysine methyltransferases that regulate expression of diverse genes. Aberrant activation of MLL promotes leukemia as well as solid tumors in humans, highlighting the urgent need for the development of an MLL inhibitor. We screened and isolated MLL1-binding ssRNAs using SELEX (Systemic Evolution of Ligands by Exponential enrichment) technology. When sequences in sub-libraries were obtained using next-generation sequencing (NGS), the most enriched aptamers—APT1 and APT2—represented about 30% and 26% of sub-library populations, respectively. Motif analysis of the top 50 sequences provided a highly conserved sequence: 5′-A[A/C][C/G][G/U][U/A]ACAGAGGG[U/A]GG[A/C] GAGUGGGU-3′. APT1, APT2, and APT5 embracing this motif generated secondary structures with similar topological characteristics. We found that APT1 and APT2 have a good binding activity and the analysis using mutated aptamer variants showed that the site information in the central region was critical for binding. In vitro enzyme activity assay showed that APT1 and APT2 had MLL1 inhibitory activity. Three-dimensional structure prediction of APT1-MLL1 complex indicates multiple weak interactions formed between MLL1 SET domain and APT1. Our study confirmed that NGS-assisted SELEX is an efficient tool for aptamer screening and that aptamers could be useful in diagnosis and treatment of MLL1-mediated diseases.
Aptamers, Nucleotide ; Conserved Sequence ; Diagnosis ; Histones ; Humans ; In Vitro Techniques ; Leukemia ; Ligands ; Lysine ; Mass Screening ; Methyltransferases ; Myeloid-Lymphoid Leukemia Protein ; RNA