BACKGROUND: T cell dysfunction, which includes exhaustion, anergy, and senescence, is a distinct T cell differentiation state that occurs after antigen exposure. Although T cell dysfunction has been a cornerstone of cancer immunotherapy, its potential in transplant research, while not yet as extensively explored, is attracting growing interest. Interferon regulatory factor 4 (IRF4) has been shown to play a pivotal role in inducing T cell dysfunction. METHODS: A novel ultra-low-dose combination of Trametinib and Rapamycin, targeting IRF4 inhibition, was employed to investigate T cell proliferation, apoptosis, cytokine secretion, expression of T-cell dysfunction-associated molecules, effects of mitogen-activated protein kinase (MAPK) and mammalian target of rapamycin (mTOR) signaling pathways, and allograft survival in both in vitro and BALB/c to C57BL/6 mouse cardiac transplantation models. RESULTS: In vitro , blockade of IRF4 in T cells effectively inhibited T cell proliferation, increased apoptosis, and significantly upregulated the expression of programmed cell death protein 1 (PD-1), Helios, CD160, and cytotoxic T lymphocyte-associated antigen (CTLA-4), markers of T cell dysfunction. Furthermore, it suppressed the secretion of pro-inflammatory cytokines interferon (IFN)-γ and interleukin (IL)-17. Combining ultra-low-dose Trametinib (0.1 mg·kg -1 ·day -1 ) and Rapamycin (0.1 mg·kg -1 ·day -1 ) demonstrably extended graft survival, with 4 out of 5 mice exceeding 100 days post-transplantation. Moreover, analysis of grafts at day 7 confirmed sustained IFN regulatory factor 4 (IRF4) inhibition, enhanced PD-1 expression, and suppressed IFN-γ secretion, reinforcing the in vivo efficacy of this IRF4-targeting approach. The combination of Trametinib and Rapamycin synergistically inhibited the MAPK and mTOR signaling network, leading to a more pronounced suppression of IRF4 expression. CONCLUSIONS Targeting IRF4, a key regulator of T cell dysfunction, presents a promising avenue for inducing transplant immune tolerance. In this study, we demonstrate that a novel ultra-low-dose combination of Trametinib and Rapamycin synergistically suppresses the MAPK and mTOR signaling network, leading to profound IRF4 inhibition, promoting allograft acceptance, and offering a potential new therapeutic strategy for improved transplant outcomes. However, further research is necessary to elucidate the underlying pharmacological mechanisms and facilitate translation to clinical practice.
Animals ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Interferon Regulatory Factors/metabolism* ; Heart Transplantation/methods* ; T-Lymphocytes/immunology* ; Sirolimus/therapeutic use* ; Pyridones/therapeutic use* ; Graft Survival/drug effects* ; Pyrimidinones/therapeutic use* ; Cell Proliferation/drug effects* ; Apoptosis/drug effects* ; Male ; Signal Transduction/drug effects*

Background::Homoharringtonine (HHT) is an effective anti-inflammatory, anti-viral, and anti-tumor protein synthesis inhibitor that has been applied clinically. Here, we explored the therapeutic effects of HHT in a mouse heart transplant model.Methods::Healthy C57BL/6 mice were used to observe the toxicity of HHT in the liver, kidney, and hematology. A mouse heart transplantation model was constructed, and the potential mechanism of HHT prolonging allograft survival was evaluated using Kaplan–Meier analysis, immunostaining, and bulk RNA sequencing analysis. The HHT-T cell crosstalk was modeled ex vivo to further verify the molecular mechanism of HHT-induced regulatory T cells (Tregs) differentiation. Results::HHT inhibited the activation and proliferation of T cells and promoted their apoptosis ex vivo. Treatment of 0.5 mg/kg HHT for 10 days significantly prolonged the mean graft survival time of the allografts from 7 days to 48 days ( P <0.001) without non-immune toxicity. The allografts had long-term survival after continuous HHT treatment for 28 days. HHT significantly reduced lymphocyte infiltration in the graft, and interferon-γ-secreting CD4 + and CD8 + T cells in the spleen ( P <0.01). HHT significantly increased the number of peripheral Tregs (about 20%, P <0.001) and serum interleukin (IL)-10 levels. HHT downregulated the expression of T cell receptor (TCR) signaling pathway-related genes ( CD4, H2-Eb1, TRAT1, and CD74) and upregulated the expression of IL-10 and transforming growth factor (TGF) -β pathway-related genes and Treg signature genes ( CTLA4, Foxp3, CD74, and ICOS). HHT increased CD4 + Foxp3 + cells and Foxp3 expression ex vivo, and it enhanced the inhibitory function of inducible Tregs. Conclusions::HHT promotes Treg cell differentiation and enhances Treg suppressive function by attenuating the TCR signaling pathway and upregulating the expression of Treg signature genes and IL-10 levels, thereby promoting mouse heart allograft acceptance. These findings may have therapeutic implications for organ transplant recipients, particularly those with viral infections and malignancies, which require a more suitable anti-rejection medication.

Objective:To investigate the biological effects of acute high-dose radon exposure on mice.Methods:BALB/c male mice aged 6 to 8 weeks were exposed once in an HD-3 ecological radon chamber with an average radon concentration of 7 × 10 5 Bq/m 3 for 10 h. Mice were weighed, their lung tissues and blood samples were collected at 1, 2 and 3 months after exposure. Control groups were set up at the three time points with four mice in each group. For these mice, the lung tissue pathology was observed using hematoxylin-eosin (HE) staining method, routine blood tests were conducted using a hematology analyzer and the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in the serum and lung tissues were measured using corresponding assay kits. Results:The HE staining result revealed that compared to the control groups, the experimental groups exhibited thickening of alveolar walls and increased infiltration of granulocyte, whose degrees, however, reduced over time and displayed no significant difference at 3 months after exposure. There was no significant difference in body weight or blood routine between the experimental and control groups. The detection result revealed decreased SOD levels in the lung tissues at 2 months after exposure, which were (11.34 ± 1.03) U/mgprot and (9.75 ± 0.71) U/mgprot, respectively for the control and experimental groups ( t = 2.54, P < 0.05). The MDA levels in lung tissue increased at 1 month after exposure, which were(2.30 ± 0.24) and (2.77 ± 0.29) nmol/mgprot, respectively for the control and experimental groups ( t = 2.49, P < 0.05). At 3 months after exposure, the SOD and MDA levels differed insignificantly between the control and experimental groups ( P > 0.05). Conclusions:After acute high-dose radon exposure, the mice suffered damage to the lung tissue, with changes in their oxidative stress indicators being detected. However, these effects gradually diminished at 3 months after exposure. Additionally, acute high-dose radon exposure did not give rise to significant changes in the body weight or routine blood result of the mice.

Objective:To evaluate the effect of dexmedetomidine on pyroptosis in rats with endotoxin-induced acute lung injury (ALI).Methods:Sixty SPF male Sprague-Dawley rats, aged 6 weeks, weighing 200-220 g, were divided into 5 groups ( n=10 each) by a random number table method: control group (group C), ALI group and different doses of dexmedetomidine groups (D 1-3 groups). In ALI group and D 1-3 groups, LPS 5 mg/kg was intraperitoneally injected to establish endotoxin-induced ALI model.Immediately after establishing the model, dexmedetomidine 12.5, 25.0 and 50.0 μg/kg were intraperitoneally injected in D 1-3 groups, and the equal volume of normal saline was intraperitoneally injected in group C, once a day for 14 consecutive days.After the end of administration, the rats were sacrificed, the left bronchus was lavaged, and the left bronchoalveolar lavage fluid (BALF) was collected for determination of the concentrations of interleukin-1beta (IL-1β), IL-6 and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay), and the lung tissue was taken for determination of the wet/dry weight ratio (W/D ratio) and expression of cleaved-caspase-1, N-terminal of the spliceosome (GSDMD-N), IL-18 and IL-1β (by Western blot) and for microscopic examination of the pathological changes (with a light microscope). Results:Compared with group C, the W/D ratio of lung tissues was significantly increased, the concentrations of IL-1β, IL-6 and TNF-α in BALF were increased, the expression of cleaved-caspase-1, GSDMD-N, IL-1β and IL-18 in lung tissues was up-regulated ( P<0.05), and the pathological damage was aggravated in ALI group and D 1-3 groups.Compared with group ALI, the W/D ratio of lung tissues was significantly decreased, and the concentrations of IL-1β, IL-6 and TNF-α in BALF were decreased, the expression of cleaved-caspase-1, GSDMD-N, IL-1β and IL-18 in lung tissues was down-regulated in a dose-dependent manner ( P<0.05), and the pathological damage was significantly reduced in D 1-3 groups. Conclusion:The mechanism by which dexmedetomidine attenuates endotoxin-induced ALI may be related to inhibition of pyroptosis and reduction of inflammatory responses in rats.

Objective: To assess the outcomes of partial cricotracheal resection (CTR) and extended cricotracheal resection (ECTR) for severe laryngotracheal stenosis. Methods: From November 2009 to September 2017, 18 patients underwent CTR and ECTR at the Department of Otorhinolaryngology Head and Neck Surgery, Tangdu Hospital, Air Force Medical University for severe laryngotracheal stenosis were reviewed retrospectively. There were 12-male and 6-female patients, with the age ranged from 4 to 56 years (median 25 years). The causes were postintubation in 11 cases, cervical trauma in 4, idiopathic in 3. The stenosis located in subglottic and tracheal (n=12), glottic and subglottic and tracheal (n=3), subglottic (n=2), and glottic and subglottic (n=1). Two patients had concurrent unilateral vocal cord palsy.One patient had undergone previous endoscopic balloon dilation and 8 patients had previous laryngotracheal reconstruction. The stenosis was graded according to modified Myer-Cotton classification as follows: Ⅲb (n=1), Ⅲc(n=1), Ⅳa (n=2), Ⅳb (n=12), Ⅳc (n=2). The surgical outcomes and complications were recorded. Results: Among 18 patients,11 of the 12 patients undergoing CTR were decannulated. Five of the 6 patients undergoing ECTR were decannulated. Resected airway length ranged from 1.5 to 4.0 cm (median 2.8 cm). Surgical complications included infection of incision wound in 2 cases, anastomotic granulation in 2, cervical subcutaneous emphysema in 1, aspiration in 1, and unilateral arytenoid prolapse in 1. No recurrent laryngeal nerve injury or tracheoesophageal fistula occurred. The median follow up was 11 months. Conclusions CTR is efficient for severe subglottic and upper tracheal stenosis while ECTR is efficient for subglottic stenosis extended to the glottis. Both procedures also provide a salvage therapy for patients with previous failed treatments.

The aim of this study is to o explore the diagnosis, treatment and prognosis of idiopathic subglottic stenosis （ISS） The clinical data of 15 patients with idiopathic subglottic stenosis treated in our department were analyzed retrospectively. The degree of stenosis was classified by the Cotton Airway grading system of Myer, with 8 cases of gradeⅡ, 4 cases of grade Ⅲ and 3 cases of grade Ⅳ. The time of follow-up of HTSS was 0.5-10 years. All 15 patients were successfully extubated without asphyxia, decannulation and wound nonunion. For patients with idiopathic subglottic stenosis in the non-progressive stage, active surgical treatment strategy should be adopted and treated individually. The prognosis is satisfactory.

Objective: The aim of this study is to o explore the diagnosis, treatment and prognosis of idiopathic subglottic stenosis （ISS） Method: The clinical data of 15 patients with idiopathic subglottic stenosis treated in our department were analyzed retrospectively. The degree of stenosis was classified by the Cotton Airway grading system of Myer, with 8 cases of gradeⅡ, 4 cases of grade Ⅲ and 3 cases of grade Ⅳ. Result: The time of follow-up of HTSS was 0.5-10 years. All 15 patients were successfully extubated without asphyxia, decannulation and wound nonunion. Conclusion For patients with idiopathic subglottic stenosis in the non-progressive stage, active surgical treatment strategy should be adopted and treated individually. The prognosis is satisfactory.

Objectives: . The simulation of microlaryngeal skills is rarely seen in surgical training, but it is particularly important in phonomicrosurgery. This study described and validated the laryngeal surgical simulator through surgical training. Methods: . A simple and low-cost simulator was developed for the fixation of the suspension laryngoscope and porcine larynges. Twenty participants with work skills and experience did preparation before training, and performed suture and carbon dioxide (CO2) laser cordectomy for simulator evaluation. The results were proposed by the aspects of time taken for each procedure, the global rating scale, a procedure-specific assessment, and a post-simulation questionnaire. Results: . All participants completed the preparation within 9 minutes and reached the conclusion that the microlaryngeal surgical simulator was helpful in improving their surgical skills. The performance of experts was superior to that of novices in both suture and CO2 laser cordectomy. Conclusion . This simulator could be easily assembled and was successfully validated by microlaryngeal surgical training both subjectively and objectively. It may be helpful to clinicians in microlaryngeal skills.

Objective: To explore the application value of thyroid alar cartilage(TAC) in the laryngotracheal reconstruction of subglotticstenosis in the paediatric population. Methods: Twelve patients(7 males,5 females; range from 2.3 to 12.0 years) with subglotticstenosis who had undergone laryngotracheal reconstruction procedures at our hospital fromSeptember 2016 to July 2018 were analyzed retrospectively.The degree of stenosis was classified according to Myer-Cotton classification system:grade Ⅱ(n=4),grade Ⅲ(n=6) and grade Ⅳ(n=2). The stenosis planes were subglotticbut did not exceed the 3rd tracheal cartilage ring. Results: Of all 12 patients, 11 were decannulated and 1 failed. All patients with grade Ⅱ and Ⅲ patients were decannulated at one procedure. There were no severe complications such as tube dislogement,asphyxia and unhealed wound. Conclusions The TAC for widening laryngotracheal lumen is relatively simple and reliable for laryngotracheal reconstruction in the pediatric population with subglottic stenosis. This would′t cause any structural damage to the laryngeal cavity.

BACKGROUND: The purpose of this study is to compare the survival time of non-small cell lung cancer (NSCLC) patients with different organ metastasis. Among all cancers, the morbidity and mortality of lung cancer is the highest worldwide, which may caused by local recurrence and distant metastasis, and the location of metastasis may predict the prognosis of patients. METHODS: A total of 117,542 patients with NSCLC diagnosed between 2010 and 2014 were enrolled from Surveillance, Epidemiology, and End Result (SEER) databases, and the relationship between distant metastasis and survival time was retrospectively analyzed. RESULTS: Of all the 117,542 patients diagnosed with non-small cell lung cancer, 42,071 (35.8%) patients had different degrees of distant metastasis during their medical history, including 26,932 single organ metastases and 15,139 multiple organ metastases, accounting for 64.0% and 36.0% of the metastatic patients respectively. Compared with patients with no metastasis, whose median survival time was 21 months, the median survival time of patients with metastases was 7 months (lung), 6 months (brain), 5 months (bone), 4 months (liver), and 3 months (multiple organ) respectively, and the difference was significant (P<0.001, except liver vs multiple organ P=0.650); Most patients with NSCLC (88.4%) eventually died of lung cancer. CONCLUSIONS Distant metastasis of NSCLC patients indicates poor prognosis. In NSCLC patients with single organ metastasis, the prognosis of lung metastasis is the best, and liver metastasis is the worst, and multiple organ metastasis is worse than single organ metastasis.
Aged ; Aged, 80 and over ; Bone Neoplasms ; mortality ; secondary ; Brain Neoplasms ; mortality ; secondary ; Carcinoma, Non-Small-Cell Lung ; mortality ; pathology ; Female ; Humans ; Liver Neoplasms ; mortality ; secondary ; Lung Neoplasms ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Prognosis ; Retrospective Studies