中南大学学报(医学版)最新文献

Objectives: IgA nephropathy (IgAN) is the most common primary glomerular disease in China, but its pathogenesis remains unclear. This study aims to explore the regulatory role of the mammalian target of rapamycin (mTOR) signaling pathway in autophagy and mesangial proliferation during renal injury in IgA.

Methods: The activity of mTOR and autophagy was evaluated in kidney samples from IgAN patients and in an IgAN mouse model induced by oral bovine serum albumin and carbon tetrachloride (CCl4) injection. mTOR inhibitors (rapamycin) and activators [bpV(phen)] were administered to the IgAN mouse model to observe the effects of mTOR on autophagy and renal lesions. In human mesangial cells treated with polymeric IgA1 (p-IgA1) and mTOR modulators, the expression and distribution of cell cycle proteins were assessed, along with the effects of mTOR on mesangial cell proliferation and autophagy.

Results: Increased mTOR activity and decreased autophagy were observed in kidney tissues from IgAN patients and the mouse model, as evidenced by elevated phosphorylated mTOR (p-mTOR) levels and reduced LC3 expression. In the IgAN mouse model, rapamycin inhibited mTOR, restored autophagy, reduced mesangial IgA deposition, alleviated mesangial cell proliferation, and decreased proteinuria (all P<0.05). In contrast, bpV(phen) activated mTOR, further suppressed autophagy, exacerbated kidney damage, and increased proteinuria (all P<0.05). In vitro, p-IgA1 induced mesangial cell proliferation and inhibited autophagy, effects that were reversed by rapamycin and aggravated by bpV(phen) (all P<0.05). mTOR regulated mesangial cell proliferation by altering cell cycle distribution, with rapamycin inducing G1 phase arrest and bpV(phen) promoting cell cycle progression. Additionally, cyclinD1 expression in renal cortex was up-regulated in the IgAN mouse model, further increased by bpV(phen), and reduced by rapamycin (all P<0.05).

Conclusions: Inhibition of the mTOR signaling pathway enhances renal autophagy, reduces mesangial cell proliferation, and improves renal injury in IgAN.

OTU domain-containing protein 3 (OTUD3) is a crucial deubiquitinase that exhibits significant expression differences across various disease models. OTUD3 plays a role in regulating biological functions such as apoptosis, inflammatory responses, cell cycle, proliferation, and invasion in different cell types. By deubiquitinating key substrate proteins, OTUD3 is involved in essential physiological and pathological processes, including innate antiviral immunity, neural development, neurodegenerative diseases, and cancer. OTUD3 exhibits tumor-suppressive effects in breast cancer, esophageal cancer, colon cancer, and papillary thyroid cancer, but acts as an oncogenic in liver and lung cancers. OTUD3 serves as a biomarker in predicting diagnosing, and assessing prognosis for certain malignancies. Despite its potential, the molecular mechanisms of OTUD3 in many diseases are still not well-understood, and exploring OTUD3's regulatory mechanisms is essential for comprehending its roles in immunity and disease. Future research will focus on developing OTUD3-targeted therapies.

Objectives: Software for pharmacological modeling and statistical analysis is essential for drug development and individualized treatment modeling. This study aims to develop a pharmacokinetic analysis cloud platform that leverages cloud-based benefits, offering a user-friendly interface with a smoother learning curve.

Methods: The platform was built using Rails as the framework, developed in Julia language, and employs PostgreSQL 14 database, Redis cache, and Sidekiq for asynchronous task management. Four commonly used modules in clinical pharmacology research were developed: Non-compartmental analysis, bioequivalence/bioavailability analysis, compartment model analysis, and population pharmacokinetics modeling. The platform ensured comprehensive data security and traceability through multiple safeguards, including data encryption, access control, transmission encryption, redundant backups, and log management. The platform underwent basic function, performance, reliability, usability, and scalability testing, along with practical case studies.

Results: The CPhaMAS cloud platform successfully implemented the 4 module functionalities. The platform provides a list-based navigation for users, featuring checkbox-style interactions. Through cloud computing, it allows direct online data analysis, saving computer storage and minimizing performance requirements. Modeling and visualization do not require programming knowledge. Basic functionality achieved 100% completion, with an average annual uptime of over 99%. Server response time was between 200 to 500 ms, and average CPU usage was maintained below 30%. In a practical case study, cefotaxime sodium/tazobactam sodium injection (6꞉1 ratio) displayd near-linear pharmacokinetics within a dose range of 1.0 to 4.0 g, with no significant effect of tazobactam on the pharmacokinetic parameters of cefotaxime, validating the platform's usability and reliability.

Conclusions: CPhaMAS provides an integrated modeling and statistical tool for educators, researchers, and industrial professionals, enabling non-compartmental analysis, bioequivalence/bioavailability analysis, compartmental model building, and population pharmacokinetic modeling and simulation.

Objectives: Islet transplantation is one of the most promising curative methods for type 1 diabetes mellitus (T1DM), but early hypoxic death of the graft post-transplantation impedes successful treatment. To improve the efficacy of islet transplantation and enhance islet cell resistance to hypoxia, reducing hypoxic injury before revascularization is crucial. Mesenchymal stem cells (MSCs) are known to regulate immune responses and protect against hypoxic damage through paracrine mechanisms. This study aims to verify the protective effects of MSC-conditioned medium (CM) in enhancing islet cells' tolerance to hypoxic conditions and preserving islet graft function.

Methods: MIN6 cells were cultured under hypoxic conditions (1% oxygen), and their viability was assessed at different time points using AO/PI staining, observed through fluorescence microscopy. MIN6 cells were treated with varying concentrations of MSC-CM under normal and hypoxic conditions. At different time points, cell viability was measured by Annexin/PI flow cytometry, and insulin secretion capacity was assessed through glucose-stimulated insulin secretion tests. A NCG T1DM mouse model was established, and islet cells from BALB/c mice were co-incubated with MSC-CM for 24 hours. The islet cells were then transplanted under the renal capsule of NCG T1DM mice. Mice body weight and blood glucose levels were monitored, and glucose tolerance tests were conducted to evaluate graft function. Graft survival was further assessed by HE staining and insulin immunohistochemistry.

Results: Under hypoxic conditions, MIN6 cell death increased with prolonged hypoxia. Flow cytometry showed that after 48 hours of hypoxia, the survival rate of MIN6 cells was significantly lower than that of the normoxic group [(68.07±7.90)% vs (94.57±2.12)%, P<0.01)]. MSC-CM treatment restored the insulin secretion function of MIN6 cells under hypoxia, with the stimulation index (SI) increasing from 1.43±0.06 to 1.77±0.02 (P<0.001). Both 10% and 20% MSC-CM effectively mitigated hypoxic damage, whereas 30% MSC-CM had weaker effects. Glucose-stimulated insulin secretion results showed trends consistent with cell survival. Primary mouse islet cells pretreated with 10% MSC-CM and transplanted under the renal capsule of T1DM mice showed a sustained decrease in blood glucose levels 5 days post-surgery. HE staining and insulin immunohistochemistry indicated that the islet cells in the MSC-CM group maintained more intact morphology and higher insulin secretion. Glucose tolerance tests demonstrated better graft function in the MSC-CM group.

Conclusions: Hypoxia significantly reduces the survival of MIN6 cells and suppresses their insulin secretion function. However, MSC-CM can significantly improve hypoxia-induced cell death and functional decline, and protect islet graft function in a T1DM mouse transplantation model.

Immunotherapy has led to groundbreaking advances in anti-tumor treatment, yet significant clinical challenges remain such as the low proportion of beneficiaries and the lack of effective platforms for predicting therapeutic response. Organoid technology provides a novel solution to these issues. Organoids are three-dimensional tissue cultures derived from adult stem cells or pluripotent stem cells that closely replicate the structural and biological characteristics of native organs, demonstrating particularly strong potential in modeling the tumor microenvironment (TME). Tumor organoids can simulate TME effectively by retaining endogenous matrix components, including various immune cells, or by adding immune cells, cancer-associated fibroblasts, and other components. This provides a novel platform for predicting immunotherapy outcomes, evaluating adoptive cell therapies, and selecting personalized treatment options for patients. Summarizing strategies for constructing tumor organoids that simulate the microenvironment and understanding their advancements in immunotherapy research and clinical application can provide new insights for the development of tumor immunotherapy.

Objectives: Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii (T. gondii), which can lead to complications such as encephalitis and ocular toxoplasmosis. The disease becomes more severe when the host's immune system is compromised. Rhoptry proteins are major virulence factors that enable T. gondii to invade host cells. This study aims to construct a T. gondii rhoptry protein 41 (rop41/ROP41) gene knockout strain and preliminarily investigate the biological function of rop41.

Methods: Using CRISPR/Cas9 technology, a specific single-guide RNA (sgRNA) for the target gene was designed and linked to a recombinant plasmid. Homologous fragments were fused with a pyrimethamine resistance gene for selection purposes. The recombinant plasmid and the homologous fragments were electroporated into T. gondii, and PCR identification was performed after drug selection and monoclonal screening. Plaque assays were used to comprehensively assess whether rop41 affected the growth and proliferation of T. gondii in host cells. Invasion and proliferation assays were conducted to evaluate the invasion ability of the knockout strain into host cells and its intracellular proliferation capacity. The STRING database was utilized to construct a protein-protein interaction (PPI) network, and functional enrichment analysis was performed to predict the signaling pathways in which ROP41 might be involved.

Results: The T. gondiirop41 gene knockout strain (RH Δku80Δrop41) was successfully constructed and stably inherited. Plaque assays showed that compared with the parental strain, the number of plaques formed by the rop41 gene knockout strain did not significantly decrease, but the reduction in plaque size was statistically significant (P<0.05). After the rop41 gene was knocked out, the invasion ability of T. gondii was reduced, but there was no statistically significant difference in its proliferation ability (P>0.05). The PPI network revealed that ROP41 was associated with other protein kinases and autophagy-related proteins. Enrichment analysis indicated that proteins interacting with ROP41 may be involved in signal transduction, biosynthesis, metabolism, and autophagy-related pathways and could be components of various kinase complexes and phagocytic vesicles.

Conclusions: The T. gondii RH Δku80Δrop41 strain has been successfully constructed. ROP41 primarily affects the ability of T. gondii to invade host cells and may play a role in signal transduction and autophagy-related pathways between T. gondii and the host.

Objectives: Thalamic hematoma patients present with diverse clinical conditions, and treatment approaches vary widely. Currently, the use of disposable portable endoscope surgery has been rapidly adopted in many hospitals, but outcomes can vary significantly. Surgical approaches and techniques for thalamic hematoma often reference those used for basal ganglia hemorrhage, but their effectiveness remains uncertain. This study aims to explore the advantages of using disposable portable endoscopes in removing thalamic hematoma via the superior parietal lobule, providing guidelines for clinicians to manage thalamic bleeding effectively.

Methods: Clinical data of patients with thalamic hematoma who underwent either disposable portable endoscope or microscope surgery at the Third Xiangya Hospital, Central South University, were retrospectively analyzed. Surgical duration, hematoma clearance rate, length of hospital stay, improvement rate in Glasgow Coma Scale (GCS) score at 24 hours post-operation, and incidence of pulmonary infection were compared between the 2 groups.

Results: Compared with the microscope group, the disposable portable endoscope group had shorter operation time, higher hematoma clearance rate, shorter hospital stay, and lower incidence of pulmonary infection (all P<0.05). However, there was no significant difference in the improvement rate of GCS score at 24 hours post-operation between the 2 groups (P>0.05).

Conclusions: In the surgical removal of thalamic hematoma via the superior parietal lobule, the disposable portable endoscope offers advantages such as shorter surgical duration, better visualization, higher hematoma clearance rate, improved surgical efficiency, shorter hospital stay, and lower incidence of pulmonary infection. Therefore, it can be considered as a preferred surgical treatment option for patients with thalamic hematoma.

Objectives: Sleep quality in kidney transplant recipients is closely associated with symptoms of fatigue and depression. Although subjective assessment tools like the Pittsburgh Sleep Quality Index and the Richards-Campbell Sleep Questionnaire (RCSQ) are widely used to evaluate sleep quality, there is a lack of studies utilizing polysomnography for objective evaluation. This study aims to investigate the correlation between sleep quality, fatigue, and depression in kidney transplant recipients using both subjective and objective methods, providing scientific evidence for improving their quality of life.

Methods: The cross-sectional study conveniently sampled 50 kidney transplant recipients from a transplant center in a general hospital between August 2018 and March 2020. Subjective and objective sleep parameters were evaluated using the RCQS and polysomnography, respectively. The Fatigue Severity Scale was used to assess fatigue, and the Hamilton Depression Scale was employed to measure depression levels.

Results: A lower proportion of rapid eye movement (REM) sleep was associated with increased fatigue. Additionally, higher wake time percentages and poorer sleep quality were significantly correlated with greater depression severity.

Conclusions: This study underscores the critical importance of effectively managing sleep quality in kidney transplant recipients and addressing their fatigue and depression symptoms. These findings lay a foundation for developing targeted nursing and therapeutic strategies.