Glutamine supplementation improves the activity and immunosuppressive action of induced regulatory T cells in vitro and in vivo.

BACKGROUND: Glutamine is crucial for the activation and efficacy of T cells, and may play a role in regulating the immune environment. This study aimed to investigate the potential role of glutamine in the activation and proliferation of induced regulatory T cells (iTregs). METHODS: CD4+CD45RA+T cells were sorted from peripheral blood mononuclear cells and cultured to analyze iTreg differentiation. Glutamine was then added to the culture system to evaluate the effects of glutamine on iTregs by determining oxidative phosphorylation (OXPHOS), apoptosis, and cytokine secretion. Additionally, a humanized murine graft-versus-host disease (GVHD) model was constructed to confirm the efficacy of glutamine-treated iTregs in vivo. RESULTS: After being cultured in vitro, glutamine significantly enhanced the levels of Foxp3, CTLA-4, CD39, CD69, IL-10, TGF-ß, and Ki67 (CTLA-4, IL-10, TGF-ß are immunosuppressive markers of iTregs) compared with that of the control iTregs (P < 0.05). Furthermore, the growth curve showed that the proliferative ability of glutamine-treated iTregs was better than that of the control iTregs (P < 0.01). Compared with the control iTregs, glutamine supplementation significantly increased oxygen consumption rates and ATP production (P < 0.05), significantly downregulated Annexin V and Caspase 3, and upregulated BCL2 (P < 0.05). However, GPNA significantly reversed the effects of glutamine (P < 0.05). Finally, a xeno-GVHD mouse model was successfully established to confirm that glutamine-treated iTregs increased the mice survival rate, delayed weight loss, and alleviated colon injury. CONCLUSION: Glutamine supplementation can improve the activity and immunosuppressive action of iTregs, and the possible mechanisms by which this occurs are related to cell proliferation, apoptosis, and OXPHOS.

The lncRNA TRG-AS1 promotes the growth of colorectal cancer cells through the regulation of P2RY10/GNA13.

BACKGROUND: The lncRNA TRG-AS1 and its co-expressed gene P2RY10 are important for colorectal cancer (CRC) occurrence and development. The purpose of our research was to explore the roles of TRG-AS1 and P2RY10 in CRC progression. METHODS: The abundance of TRG-AS1 and P2RY10 in CRC cell lines (HT-29 and LoVo) and normal colon cells FHC was determined and difference between CRC cells and normal cells was compared. LoVo cells were transfected with si-TRG-AS1 and si-P2RY10 constructs. Subsequently, the viability, colony formation, and migration of the transfected cells were analyzed using cell counting kit-8, clonogenicity, and scratch-wound/Transwell® assays, respectively. Cells overexpressing GNA13 were used to further explore the relationship between TRG-AS1 and P2RY10 along with their downstream functions. Finally, nude mice were injected with different transfected cell types to observe tumor formation in vivo. RESULTS: TRG-AS1 and P2RY10 were significantly upregulated in HT-29 and LoVo compared to FHC cells. TRG-AS1 knockdown and P2RY10 silencing suppressed the viability, colony formation, and migration of LoVo cells. TRG-AS1 knockdown downregulated the expression of P2RY10, GNA12, and GNA13, while P2RY10 silencing downregulated the expression of TRG-AS1, GNA12, and GNA13. Additionally, GNA13 overexpression reversed the cell growth and gene expression changes in LoVo cells induced by TRG-AS1 knockdown or P2RY10 silencing. In vivo experiments revealed that CRC tumor growth was suppressed by TRG-AS1 knockdown and P2RY10 silencing. CONCLUSIONS: TRG-AS1 knockdown repressed the growth of HT-29 and LoVo by regulating P2RY10 and GNA13 expression.

A comparison of the therapeutic outcomes between primary duct closure and T-tube drainage after laparoscopic common bile duct exploration: a single-centre retrospective study.

Introduction: In emergency surgery for acute obstruction of the common bile duct (CBD), primary duct closure (PC) of the CBD after laparoscopic common bile duct exploration (LCBDE) remains challenging. Aim: To explore the safety and effectiveness of this surgical method after LCBDE in patients with acute choledocholithiasis and discuss the feasibility of PC in the CBD. Material and methods: This retrospective study on surgical efficacy and safety involved 232 patients treated at The Third Affiliated Hospital of Soochow University between January 2015 and December 2019. These patients underwent LC + LCBDE for acute choledocholithiasis and were categorized into PC and T-tube drainage (TD) groups based on the method of closure of the CBD. The basic preoperative information, intraoperative situation, postoperative situation, and complications were analysed and compared between groups. Results: The baseline characteristics and preoperative information of patients between the 2 groups were balanced. Patients in the PC group had a shorter operation time (p < 0.001) and CBD suturing time (p < 0.001) than those in the TD group. In addition, compared with the TD group in postoperative situations, gastrointestinal recovery (p = 0.002), drainage removal (p < 0.001), and the length of postoperative hospital stay (p = 0.004) were markedly decreased in the PC group. In terms of intraoperative blood loss (p = 0.961), use of pipe washing (49.0 vs. 54.6%, p = 0.397), use of stone basket (50.0 vs. 42.3%, p = 0.243), use of electrohydraulic lithotripsy (1.0 vs. 3.1%, p = 0.525), postoperative liver function, and complications there was no significant difference between the PC and TD groups. No intraoperative transfusion and postoperative mortality occurred in either group. During 6 months of follow-up, only 1 patient showed biliary stricture in the PC group, and 2 and 4 patients in the PC and TD groups, respectively, showed residual stones. Conclusions: PC after LCBDE in acute choledocholithiasis patients displays better therapeutic outcomes than TD in some intraoperative and postoperative situations. PC of the CBD after LCBDE is a safe and effective therapeutic option in acute choledocholithiasis patients.

Therapeutic efficacy of programmed spatial anatomy of the myopectineal orifice in total extraperitoneal hernioplasty: a retrospective study.

This study aimed to investigate the therapeutic efficacy of programmed spatial anatomy of myopectineal orifice technique in laparoscopic total extraperitoneal hernioplasty (TEP) surgery. A total of 121 adult male patients with unilateral inguinal hernias who underwent TEP in the Department of General Surgery, Wujin Hospital, affiliated with Jiangsu University, from January 2019 to December 2020 were selected. Patients were divided into the procedural (63 cases) and traditional groups (58 cases) according to the surgical methods adopted. The procedural group underwent programmed spatial anatomy of the myopectineal orifice combined with TEP, and the traditional group underwent traditional TEP. The perioperative evaluation indicators and postoperative complications were observed and compared between the two groups. Compared with the traditional group, the time of handling hernia, the intraoperative operation time, intraoperative blood loss, postoperative ambulation time, and postoperative hospital stay in the procedural group were significantly reduced (P < 0.05). The incidence of postoperative complications such as sensory nerve abnormalities and chronic pain was significantly decreased (P < 0.05), and the total incidence of complications in the procedural group was significantly lower than that in the traditional group (P < 0.05). While there was no significant difference in postoperative incision infection (P > 0.05). The programmed spatial anatomy of the myopectineal orifice can significantly improve the treatment outcome of TEP, significantly improve the patients' intraoperative and postoperative indicators, and reduce the incidence of postoperative complications. It is worthy of being promoted among young physicians and basic hospitals.

CD8+iTregs mediate the protective effect of rapamycin against graft versus host disease in a humanized murine model.

CD8+Tregs are important immunoregulatory cells that participate in immunopathological processes in many diseases. Rapamycin (Rapa) is a macrolide immunosuppressant that inhibits the mammalian target of rapamycin (mTOR) and has been shown to improve CD4+-induced Tregs (iTregs) generation. This study aimed to evaluate the role of Rapa in the generation and function of CD8+iTregs. Human CD8 + CD25-CD45RA + T cells were divided into two groups, one with Rapa and the other without Rapa, and both groups were cultured under Treg-induced conditions. Rapa significantly improved Foxp3 expression and the suppressive function of CD8+iTregs in vitro. Further studies showed that Rapa suppressed inflammatory cytokine expression and enhanced anti-inflammatory cytokine expression. Under inflammatory conditions in vitro, Rapa-CD8 + iTregs sustained Foxp3 and anti-inflammatory cytokine expression. An in-depth study showed that Rapa regulated CpG demethylation in the Foxp3 region and STAT1 and STAT3 phosphorylation in CD8+iTregs. Finally, we compared the regulatory ability of Rapa and all-trans retinoic acid, another reagent that stimulates CD4+ iTreg generation in vitro, which showed that Rapa, but not all-trans retinoic acid, improved CD8+ iTreg induction and suppressed CD4+T cell expansion in vitro and protected against graft-versus-host disease in a humanized murine model in vivo. These results strongly suggest that CD8+iTregs initiated by Rapa may represent a new therapeutic strategy for inflammatory and autoimmune diseases.