Pathological Processes Research Articles

Antidepressant effect and mechanism of TMP269 on stress-induced depressive-like behavior in mice

TMP269, a class IIA histone deacetylase inhibitor with selectivity, that has a protective effect on the central nervous system, yet its specific mechanism of action remains ambiguous. Although major depressive disorder (MDD) is highly prevalent, its pathophysiology is poorly understood. Recent evidence suggests that histone deacetylase 5 plays a key role in the pathological process of depression and the fact that preclinical studies have shown HDAC5 to be a potential antidepressant target, the search for natural drugs or small molecule compounds that can target HDAC5 may be a potential therapeutic strategy for the treatment of depression. In addition, we examined the role of the Brain-derived neurotrophic factor (BDNF), an important neurotrophic factor for neuronal survival and growth, as a potential downstream target of HDAC5. We found downward revision of HDAC5 levels in the hippocampus ameliorated depressive-like behavior in LH (Learned helplessness) mice. Furthermore, injection of HDAC5 overexpressing adenoviral vectors in the hippocampal dentate gyrus of wild-type mice produced a somewhat depressive-like phenotype. Pharmacological, immunofluorescence and biochemical experiments showed that TMP269 could produce antidepressant effects by inhibiting mouse hippocampal HDAC5 and thus modulating its downstream BDNF. Over all, TMP269 mitigated LH-induced depressive-like behaviors and abnormalities in synapse formation and neurogenesis within the hippocampus. These findings suggest potential beneficial effects of TMP269 on depression.

KLF4 Inhibits the Activation of Human Hepatic Stellate Cell In Vitro.

Hepatic stellate cells (HSCs) play a crucial role in liver fibrosis. Early-stage liver fibrosis is reversible and intimately associated with the state of HSCs. Kruppel-like factor 4 (KLF4) plays a pivotal role in a wide array of physiological and pathological processes. This study aimed to investigate the effect of KLF4 on the proliferation, apoptosis and phenotype of quiescent HSCs METHODS: We designed a KLF4 lentiviral vector and a KLF4 siRNA lentiviral vector, to upregulate and silence KLF4 expression in human HSC LX-2 cells via transfection. Cell proliferation was assessed using the CCK-8 assay. Flow cytometry was used to detect the cell cycle distribution and apoptosis rate. Western blotting was used to determine the levels of some quiescence and activation markers of HSCs RESULTS: Overexpression of KLF4 significantly increased the levels of E-cadherin and ZO-1, which are quiescent HSC markers, while significantly decreased the levels of N-cadherin and a-SMA, known activated HSC markers. In contrast, cell proliferation and apoptosis rates were elevated in LX-2 cells in which KLF4 expression was silenced CONCLUSION: KLF4 inhibits the proliferation and activation of human LX-2 HSCs. It might be a key regulatory protein in the maintenance of HSC quiescence and may serve as a target for the inhibition of hepatic fibrosis.

Extracellular vesicles: Function, resilience, biomarker, bioengineering, and clinical implications

ABSTRACT Extracellular vesicles (EVs) have emerged as key players in intercellular communication, disease pathology, and therapeutic innovation. Initially overlooked as cellular debris, EVs are now recognized as vital mediators of cell-to-cell communication, ferrying a cargo of proteins, nucleic acids, and lipids, providing cellular resilience in response to stresses. This review provides a comprehensive overview of EVs, focusing on their role as biomarkers in disease diagnosis, their functional significance in physiological and pathological processes, and the potential of bioengineering for therapeutic applications. EVs offer a promising avenue for noninvasive disease diagnosis and monitoring, reflecting the physiological state of originating cells. Their diagnostic potential spans a spectrum of diseases, including cancer, cardiovascular disorders, neurodegenerative diseases, and infectious diseases. Moreover, their presence in bodily fluids such as blood, urine, and cerebrospinal fluid enhances their diagnostic utility, presenting advantages over traditional methods. Beyond diagnostics, EVs mediate crucial roles in intercellular communication, facilitating the transfer of bioactive molecules between cells. This communication modulates various physiological processes such as tissue regeneration, immune modulation, and neuronal communication. Dysregulation of EV-mediated communication is implicated in diseases such as cancer, immune disorders, and neurodegenerative diseases, highlighting their therapeutic potential. Bioengineering techniques offer avenues for manipulating EVs for therapeutic applications, from isolation and purification to engineering cargo and targeted delivery systems. These approaches hold promise for developing novel therapeutics tailored to specific diseases, revolutionizing personalized medicine. However, challenges such as standardization, scalability, and regulatory approval need addressing for successful clinical translation. Overall, EVs represent a dynamic frontier in biomedical research with vast potential for diagnostics, therapeutics, and personalized medicine.

Vasculoendothelial dysfunction and bone health in obese children: A connection with cancers

Obesity in children, a growing global epidemic, is not merely characterized by excessive body fat but also by a cascade of metabolic and functional derangements with significant musculoskeletal consequences. One of the key underpinnings of these detrimental effects is vasculoendothelial dysfunction (VED), a multifaceted pathological process characterized by impaired endothelial cell function. The endothelium, lining the vasculature, plays a pivotal role in regulating vascular tone, inflammation, and coagulation, and its dysfunction in obese children leads to a plethora of downstream complications with profound implications for musculoskeletal and bone health. Diagnosing VED in obese children remains a challenge. Traditional cardiovascular risk factors such as hypertension and dyslipidemia are often detected long after the occurrence of endothelial dysfunction. In the context of bone sarcoma, the interaction between cancer cells and vascular endothelium in the bone microenvironment is critical. The bone is a highly vascularized tissue, and the vascular endothelium plays a role in regulating bone homeostasis. In the presence of cancer cells, altered bone microenvironment provides a milieu favorable to tumor growth and invasion. The relationship between the vascular endothelium and cancer is complex and can vary depending on the type of cancer and its microenvironment. Understanding the interactions between vascular endothelium and bone sarcoma is crucial for developing targeted therapies. New molecular and cellular mechanisms involved in these interactions are awaiting to be uncovered, shedding light on potential therapeutic strategies for cancer treatment. On a relevant note, emerging vasculoendothelial markers hold promise for early detection and intervention. Early identification of VED, through the detection of vasculoendothelial markers, opens doors for timely interventions aimed at preventing its detrimental effects on musculoskeletal and bone health in obese children

Type 2 diabetes mellitus related sarcopenia: a type of muscle loss distinct from sarcopenia and disuse muscle atrophy

Muscle loss is a significant health concern, particularly with the increasing trend of population aging, and sarcopenia has emerged as a common pathological process of muscle loss in the elderly. Currently, there has been significant progress in the research on sarcopenia, including in-depth analysis of the mechanisms underlying sarcopenia caused by aging and the development of corresponding diagnostic criteria, forming a relatively complete system. However, as research on sarcopenia progresses, the concept of secondary sarcopenia has also been proposed. Due to the incomplete understanding of muscle loss caused by chronic diseases, there are various limitations in epidemiological, basic, and clinical research. As a result, a comprehensive concept and diagnostic system have not yet been established, which greatly hinders the prevention and treatment of the disease. This review focuses on Type 2 Diabetes Mellitus (T2DM)-related sarcopenia, comparing its similarities and differences with sarcopenia and disuse muscle atrophy. The review show significant differences between the three muscle-related issues in terms of pathological changes, epidemiology and clinical manifestations, etiology, and preventive and therapeutic strategies. Unlike sarcopenia, T2DM-related sarcopenia is characterized by a reduction in type I fibers, and it differs from disuse muscle atrophy as well. The mechanism involving insulin resistance, inflammatory status, and oxidative stress remains unclear. Therefore, future research should further explore the etiology, disease progression, and prognosis of T2DM-related sarcopenia, and develop targeted diagnostic criteria and effective preventive and therapeutic strategies to better address the muscle-related issues faced by T2DM patients and improve their quality of life and overall health.

Color-reversible fluorescence tracking for the dynamic interaction of SO2 with Hg2+ in living cells

Mercury ion (Hg2+) is one of the most threatening substances to human health, and the mercury poisoning can damage physiological homeostasis severely in human, even cause death. Intriguingly, Sulfur dioxide (SO2), a gas signal molecule in human, can specifically interact with Hg2+ for relieving mercury poisoning. However, the dynamic interaction of Hg2+ with SO2 at the tempospatial level and the correlation between Hg2+ and SO2 in the pathological process of mercury poisoning are still elusive. Herein, we rationally designed a reversible and dual color fluorescent probe (CCS) for dynamically visualizing Hg2+ and SO2 and deciphering their interrelationship in mercury poisoning. CCS held good sensitivity, selectivity and reversibility to Hg2+ and SO2, that enabled CCS to specifically detect SO2 and Hg2+ via cyan fluorescence channel (centered around 485 nm) and red fluorescence channel (centered around 679 nm), respectively. Notably, the separate fluorescence signal changes of CCS realized the dynamic tracing of Hg2+ and SO2 in living cells, and presented the potential for exploring the correlation between SO2 and Hg2+ in mercury poisoning.

Job burnout, cognitive functioning, and Brain-derived neurotrophic factor expression among hospital Mexican nurses.

To analyze the relationship between burnout syndrome, cognitive functions, and sBDNF (Serum Brain-derived Neurotrophic Factor) in Mexican nurses. A descriptive cross-sectional design was used. This study target staff nurses working in hospitals in Guanajuato, México. Demographic and working condition data were collected via questionnaire. The Maslach Burnout Inventory (MBI) was used to evaluate burnout. A blood sample were collected and processed by ELISA technique to measure sBDNF. Finally, the General Cognitive Assessment (CAB) of the Cognifit© neuropsychological battery was used to evaluated cognitive functions. Findings showed that there are sociodemographic characteristics and working conditions associated with burnout syndrome among nurses. Furthermore, the data demonstrated a significant decrease in sBDNF levels in burnout nurses and a negative correlation between BDNF levels and burnout syndrome. Additionally, these burnout nurse also revealed significant cognitive impairment in reasoning, memory, and attention as well as total scores of CAB. Interestingly, we found a positive correlation between sBDNF levels and the cognitive deficits in burnout nurse. Reduced BDNF levels could be a biological indicator or part of the pathological process of burnout, which could affect cognitive abilities. Reduced cognitive function in nurses has relevant implications and emphasizes the need for specialized preventive strategies because nurses make clinical decisions concerning their patients, whose situations are constantly changing.

#2983 Correlation of microRNA expression with IgA nephropathy progression: preliminary results

Abstract Background and Aims IgA nephropathy (IgAN) is the most common cause of glomerulonephritis worldwide. IgAN is diagnosed based on the result of histopathological examination. The risk factors for disease progression primarily include the level of impairment of renal function at the time of diagnosis (GFR < 60 ml/min/1.73 m2), the presence of hypertension, daily administration of more than 1.0 g, and the occurrence of hematuria. MicroRNAs (miRNAs) are important regulators of gene expression. Their role has a significant impact on the proper functioning of the body, and disorders related to their expression can lead to the development of many pathological processes in the body. The study aimed to assess the dependence of microRNA expression on the stage of the disease in patients with IgA nephropathy. Method A study was conducted involving patients newly diagnosed with IgA nephropathy based on histopathological examination results. The expression of microRNA molecules (miR-146a, miR-155, miR-200b, miR-429) was determined in urine sediment and blood. The analysis focused on examining the relationship between microRNA expression and the stage of disease progression (proteinuria, renal parameters). Blood and urine tests were performed before the start of immunosuppressive treatment with glucocorticosteroids according to the Pozzi regimen. Results Twenty-seven patients were included in the study group- 14 women (51.9%) and 13 men (48.1%). Nephrotic-range proteinuria (>3000 mg/24 h) was observed in 5 patients (18.5%), while subnephrotic proteinuria was present in 22 patients (77.8%). Reserved kidney function (eGFR > 90) was found in 17 patients (63%). The exact distribution of results according to the stage of renal failure is given in the Table 1. Statistically significant positive correlation was found between serum creatinine concentration and miR-200b expression in urine (R = 0,398, p = 0,049). Initially, no statistically significant correlation was found between the expression of selected miRNA molecules and the degree of 24-hour proteinuria. Conclusion MiRNA molecules may be promising, non-invasive biomarkers widely used in clinical practice. Determining the viability of clinical use of microRNA molecules will require a panel of miRNAs or miRNAs used in combination with other biomarkers and clinical outcomes to improve diagnostic utility.

In-depth organic mass cytometry reveals differential contents of 3-hydroxybutanoic acid at the single-cell level

Comprehensive single-cell metabolic profiling is critical for revealing phenotypic heterogeneity and elucidating the molecular mechanisms underlying biological processes. However, single-cell metabolomics remains challenging because of the limited metabolite coverage and inability to discriminate isomers. Herein, we establish a single-cell metabolomics platform for in-depth organic mass cytometry. Extended single-cell analysis time guarantees sufficient MS/MS acquisition for metabolite identification and the isomers discrimination while online sampling ensures the high-throughput of the method. The largest number of identified metabolites (approximately 600) are achieved in single cells and fine subtyping of MCF-7 cells is first demonstrated by an investigation on the differential levels of 3-hydroxybutanoic acid among clusters. Single-cell transcriptome analysis reveals differences in the expression of 3-hydroxybutanoic acid downstream antioxidative stress genes, such as metallothionein 2 (MT2A), while a fluorescence-activated cell sorting assay confirms the positive relationship between 3-hydroxybutanoic acid and target proteins; these results suggest that the heterogeneity of 3-hydroxybutanoic acid provides cancer cells with different ability to resist surrounding oxidative stress. Our method paves the way for deep single-cell metabolome profiling and investigations on the physiological and pathological processes that occur during cancer.

#2069 Elucidating the function of novel Arg1+/Clec4d+ scar-associated monocyte-derived macrophage population in driving fibrosis in kidney disease models

Abstract Background and Aims Fibrosis is the final common pathway in all progressive kidney disease. Macrophages are a major myeloid cell component of the renal mononuclear phagocyte system, with roles in defence against infection, renal injury, and repair. Using single-cell RNA sequencing, we identified a novel myeloid cell subset exclusively present in acute injury of the unilateral ureteric obstruction (UUO) model of kidney fibrosis. This population transcriptomically aligns to monocytes but is enriched for both Arginase-1 (Arg1) and the C-type lectin 4d (Clec4d) and a large number of pro-inflammatory and pro-fibrotic genes. We hypothesise that this novel Arg1+/Clec4d+ population contributes to fibrosis deposition in progressive kidney disease. Methods Monocyte-macrophage populations were characterised in different pre-clinical models; UUO, unilateral ischaemic reperfusion injury (uIRI) and subtotal nephrectomy with flow cytometry. Arg1+/Clec4d+ cells were identified by RT-qPCR, flow cytometry, and immunofluorescence. The origin and consequence of Arg1 deletion in monocytes on renal fibrosis was determined by utilising Ccr2-ERT2-TdTomato and Ccr2-ERT2-TdTomato; Arg1 fl/fl mice, respectively. Results The presence of the Arg1+/Clec4d+ cells was validated in the UUO, uIRI and subtotal nephrectomy models of kidney injury. Analysis of intra-renal inflammation revealed CD45+CD11b+Arg1+ cells persisted and increased in number across 7 days. Post-injury, Arg1+ cells were confirmed to be Clec4dhi. To confirm that Arg1+/Clec4d+ cells were derived from monocytes (Ccr2+), we administered tamoxifen to Ccr2CreERT2-TdTomato mice under a single or multiple-dose regimen following UUO surgery. At day 7 post-injury, ∼50% and ∼90% of the Arg1+ cells were TdTomato+ following a single or multiple doses of tamoxifen, respectively, suggesting that these cells are primarily monocyte derived. Moreover, the Arg1+/Clec4d+ cells were found to localise to areas of scarring (Fig. 1). To investigate the therapeutic potential of targeting Clec4d+ expressed on the Arg1+/Clec4d+ macrophages, mice underwent UUO surgery and given multiple doses of a Clec4d-neutralising antibody. A decrease in Arg1 and fibrosis-related gene expression was evident by day 7 post-injury. To understand the role of increased Arg1 expression in the cells we generated Ccr2-ERT2-TdTomato; Arg1 fl/fl mice. These mice had as expected a reduced Arg1 expression in the Ccr2+ population and metabolic profiling by SCENITH of the tissue monocytes revealed that the loss of Arg1 in the cells reduced their glucose dependence. The loss of Arg1 in these cells reduced Havcr1 gene expression in the kidney. Conclusion As fibrosis is a pathological process that can affect any organ, the work included here, in addition to the proposed future studies, has the potential to develop novel therapeutics that limit fibrosis and enhance repair to halt the progression of disease in CKD.

#2009 FGF23 in patients with diabetic kidney disease and atherosclerosis

Abstract Background and Aims Diabetes mellitus (DM) and chronic kidney disease (CKD) are risk factors for vascular atherosclerosis, a pathological process with an important inflammatory component. The coexistence of these conditions in patients with diabetic kidney disease (DKD) has additive effects with worst cardiovascular mortality and morbidity rates when compared to non-diabetic CKD patient on cardiovascular outcomes. Accumulating research has shown that increased levels of bone-derived fibroblast growth factor 23 (FGF23), the major phosphate-regulating hormone, constitute a novel predictor of cardiovascular and all-cause mortality in CKD patients, with potential effects on disorders of glucose metabolism as well. The objective of this comparative study is to determine the differences in FGF23 levels in two groups of patients with CKD, with and without DM, matched by age, sex and renal function, all of them with clinical atherosclerotic disease. We also evaluated the relationships of FGF23 with several inflammatory markers and parameters of glucose metabolism. Method Sixty-six patients with CKD stages 3-4 (33 of them with DM) and diagnosed with peripheral vascular disease were included in the study. Whole blood and serum samples were obtained from all participants. CRP values, neutrophil/lymphocyte (N/L) ratio, HbA1c (%), fasting glucose, and Triglycerides and glucose (TyG) index, among other parameters, were obtained using standardized routine methods. Circulating levels of the intact form of FGF23, tumor necrosis factor (TNF) α, interleukin (IL) 6, and IL10 were determined by ELISA immunoassays in both groups. We also performed gene expression analysis for these three cytokines in peripheral blood cells (PBCs) using qPCR. Results The mean values of eGFR, ACR and phosphate were 37.6 ± 11.2 ml/min/1.73 m2; 294.1 ± 271.2 mg/g; 3.8 ± 0.82 mg/dL; respectively without differences between patients with and without DM The median circulating FGF23 values were higher in the DM group (51.59 [37.73-89.68] vs. 38.1 [33-54.14] pg/mL; p<0.001). Likewise, patients with DM presented higher levels of TNFα both in serum concentration (1.78 [0.72-2.47] vs. 1.28 [1.01-1.88] pg/mL; p<0.01) as in the expression of mRNA in PBCs (2.1 ± 1.26 vs. 1.87 ± 1.16 a.u.; p<0.05). No differences were observed for IL6 and IL10 levels, nor for CRP concentration or N/L ratio. As expected, fasting glucose, HbA1c and TyG index were higher in the group of subjects with DM. FGF23 was inversely correlated with eGFR (r=−0.435, p<0.0001), while it showed a positive association with ACR (r=0.416, p<0.001) and, interestingly, with proinflammatory cytokines including circulating TNFα (r=0.44, p<0.001), circulating IL6 (r=0.27, p=0.04), and TNF mRNA expression in PBCs (r=0.245, p=0.047). Furthermore, circulating FGF23 was also direct and significantly correlated with HbA1c (r=0.34, p<0.01). Conclusion These results suggest that FGF23 levels are higher in DKD than in non-DM CKD patients with similar renal function. DKD patients also had higher levels of the proinflammatory cytokine TNFα. Our results also show positive correlations of FGF23 with the inflammatory cytokines TNFα and IL6, as well as with HbA1c. Previous studies, shown that elevated TNFα and glucose promote FGF23 production, which may help to explain the higher levels of FGF23 in patients with DM Future studies should clarify whether FGF23 is simply a marker of atherosclerosis disease severity or a contributing factor to this complication in CKD.

#2382 Renal tubule-derived EVs carrying complement 3 aggravate CKD vascular calcification by downregulating autophagy in vascular smooth muscle cells

Abstract Background and Aims Vascular calcification (VC) is a serious complication of chronic kidney disease (CKD) patients, recognized in KDIGO guidelines as a leading risk factor for cardiovascular diseases. Despite acknowledgment, the mechanisms of VC in CKD remain unclear. The kidneys intricately regulate blood vessel pathophysiology. Clinical studies suggest a correlation between elevated blood complement levels in CKD and arterial calcification, hinting at complement involvement in CKD-associated VC. Renal-origin complement, a significant source of circulating complement, activates in CKD. Extracellular vesicles (EVs), mediating intercellular communication, transport complement-related proteins in various physiological and pathological processes. Studies suggest that plasma EV rather than EV-depleted plasma causes pathogenic calcium salt deposition in smooth muscle cells. This study explores how renal-origin EVs carrying complement mediate CKD-associated VC. Method From October 2021 to August 2023, data from 110 clinical patients were gathered, encompassing both blood C3 levels and CT images of the thoracic aorta. Subsequently, we conducted a scoring analysis of the CT images and carried out correlation analyses. A CKD-associated VC model was induced by feeding C57BL/6j mice an adenine and high-phosphate diet for 16 weeks. Techniques, including WB, PCR, and immunofluorescence, detected complement activation and localization in renal tissues. EVs were isolated from kidney tissue and plasma and identified by NTA, electron microscopy, WB, etc. WB examined C3 and tubular markers of EVs. At the same time, the Elisa method was used to detect C3 in plasma, plasma EV, and EV-depleted plasma. Immunofluorescence investigated tubular-derived EVs uptake in calcified arteries. Tubular-derived EVs, C3a, C3aR receptor inhibitors (SB290157), and autophagy inhibitors (3-MA) intervened in high-phosphate-induced vascular smooth muscle cells (VSMCs). PCR and WB assessed calcification indicators, complement receptors, and autophagy-related indicators. Alizarin Red staining observed calcification nodules. Transcriptome sequencing was performed on control, high-phosphate, and renal tubular-derived EV intervention groups of VSMCs. An in vivo model infused renal tubular-derived EVs and injected C3aR inhibitors and autophagy inhibitors, measuring complement activation and VC indicators. Results There is a correlation between C3 and thoracic aorta calcium score in CKD patients, with a correlation coefficient of 0.3750 (P < 0.01). The CKD-associated VC mice model revealed complement C3 activation in renal tubules, secreted into the blood through EVs. The C3 proportion of plasma EV in CKD patients was significantly higher than that in the control group, but there was no statistical difference between the EV-depleted plasma control group and the CKD group. Immunofluorescence indicated tubular-derived EV uptake by the calcified vascular wall. In vitro, tubular-derived EVs enhanced VSMC osteogenic differentiation and upregulated C3aR, mimicking C3a intervention effects. C3aR receptor inhibitors produced opposing effects. Transcriptome sequencing suggested renal tubular-derived EVs downregulated VSMC autophagy-related pathways; in vitro autophagy inhibition reversed osteogenic differentiation. In vivo, reinfusing renal tubular-derived EVs exacerbated complement activation and VC, while injecting C3aR inhibitors and autophagy inhibitors alleviated CKD-associated VC. Conclusion This study confirms that tubular-derived EVs carrying C3 downregulate autophagy in VSMCs, mediating CKD-associated VC. These findings provide insights into understanding CKD-associated VC initiation and progression, revealing potential therapeutic targets.

Immunomodulatory therapy of precancers of the cervix – opinion of medical professionals and students

Objective. To assess the relevance of immunomodulatory therapy of precancerous diseases of the cervix among medical professionals and students. Precancerous diseases of the cervix combine pathological processes characterized by the structural changes in the epithelium of the cervix. If not treated, they lead to the development of cervical cancer. At present there are no selective drugs for the human papillomavirus. To administer immunotropic medicines it is necessary to know the mechanism of effect on the immune system. Materials and methods. The study was conducted using an anonymous online questionnaire on the Google Forms platform (https://vk.com/away.php?utf=1to=https%3A%2F%2Fforms.gle%2FRtT7PktnCrLqBB1x8). The questionnaire is presented as part of a one-stage (cross-sectional) study. Data analysis was carried out using Microsoft Excel 2010 graphs and tables. Empirical, sociological and statistical research methods were used. The indicators of the structure of the respondents were calculated, the reliability of the differences was determined by the Student's t-criterion, confidence differences were considered at significance of p 0.05. Results. The average age of the respondents was 30.4 ± 10.8 years. 46 people (75.4 %) are aware of the possibility of using immunomodulation as therapy for precancerous diseases of the cervix, and 15 people (24.6 %) are not aware of it. 41 respondents (67.2 %) would administer the therapy themselves and their close people; 12 people (19.7 %) find it difficult to answer; 8 respondents (13.1 %) do not agree to administer the therapy. Conclusions. Medical students have an idea about immunomodulatory therapy in gynecology in 55.9 % of cases. Doctors of different specialties in 100 % of cases are aware of the treatment, see the benefits of prescribing drugs. Students` awareness of the possibilities of immunomodulatory therapy is sufficient, they get additional knowledge from clinical recommendations on the specialty in the practical course.

#1646 Impact of sex in clinical presentation and outcomes of patients with ANCA-associated vasculitis with severe kidney disease

Abstract Background and Aims Biological sex has been associated to a more rapid rate of chronic kidney disease progression (ie, speed of kidney function loss) in male than female [1]. Testosterone can increase oxidative stress, activate the renin angiotensin system, and aggravate renal fibrosis, whereas estrogens inhibit these pathological processes in the diseased kidney [2]. Several factors such as lower disease awareness and surveillance rates as well as disparities in health-care access result in late initiation or lack of kidney replacement therapy among females [3-5]. The impact of sex in the clinical presentation and outcomes of patients with anti-neutrophil cytoplasmic antibodies (ANCA) associated vasculitis (AAV) with glomerulonephritis (AAV-GN) has not been studied, particularly in patients with severe kidney involvement at presentation (eGFR<15 mL/min/1.73 m2). Method A retrospective cohort study on MPO- or PR3-ANCA positive patients with AAV (MPA or GPA) and eGFR<15 ml/min/1.73 m2 or ESKD at presentation. Clinical presentation and outcomes (renal recovery, dialysis at presentation, and ESKD) were analyzed according with sex. Results We analyzed 166 patients with biopsy proven active AAV-GN and eGFR <15 mL/min/1.73 m2 at the time of diagnosis (Fig. 1): 78 (47%) were females and 88 (53%) were males. Arterial hypertension was more frequently present in males (85.2% vs. 70.5%, p = 0.022). Median serum creatinine (SCr) was higher in males when compared with females (5.2 [IQR 4.2-7.4] vs. 3.6 [IQR 2.8-5.1] mg/dL, p < 0.001) but there were no differences in eGFR at presentation. When analyzing only the 71 (24 females and 47 males) patients that started dialysis within 4 weeks, infections at 12 months were more common in females (75.0% vs. 36.2%, p = 0.017). The rate of dialysis initiation or progression to ESKD was not different between males (15, 62.5%) versus females (29, 61.7%) being on dialysis at 12 months. By multivariable logistic regression, factors related with dialysis initiation within 4 weeks in our cohort were SCr (OR1.478, [95% CI 1.231-1.776], p < 0.001), alveolar hemorrhage (OR 2.726, [95% CI 1.099-6.763], p = 0.031) and PR3-ANCA (OR3.155, [95% CI 1.485-6.702], p = 0.003) adjusted to sex). Patients who remained on dialysis at 12 months more often had higher SCr at diagnosis (IRR 1.098 [95% CI, 1.026–1.175], p = 0.007), severe chronicity changes on kidney biopsies (IRR 2.469 [95% CI,1.205–5.061], p = 0.014), higher chronicity score (IRR 1.178 [95%CI, 1.062-1.308], p = 0.002) and lower hemoglobin (IRR 0.824 [95%CI, 0.683-0.994], p = 0.043). Minimal chronicity changes were protective for the achievement of this endpoint (IRR 0.228 [95% CI, 0.070-0.741], p = 0.014). Multivariable Cox regression showed that patients with higher SCr at diagnosis and moderate/severe chronicity changes on biopsy were at higher risk to evolve to ESKD at 12 months compared to patients with lower chronicity grades, even when adjusted to sex. Conclusion In our cohort, male patients presented with higher SCr, and dialysis was started within 4 weeks more frequently. However, this did not result in different outcomes in terms of kidney recovery, overall dialysis rates, and progression to ESKD in patients with AAV-GN and eGFR<15 mL/min/1.73 m2 between groups.

#2305 Mice with a PAX2 missense variant are susceptible to experimental focal segmental glomerulosclerosis (FSGS) and display impaired glomerular repair

Abstract Background and Aims Focal segmental glomerulosclerosis (FSGS) is a histopathologic lesion observed in chronic kidney disease characterized by scarring of the glomerulus due to the injury and irreversible loss of highly specialized glomerular epithelial cells called podocytes. Recent studies suggest that podocytes may be regenerated by another population of glomerular epithelial cells called parietal epithelial cells (PECs), although this remains controversial. PAX2 is a transcription factor protein that regulates glomerular development and its expression persists in adult parietal epithelial cells (PECs), postulated to serve as a reservoir for podocytes in the event of injury and loss. Our team previously found that PAX2 pathogenic missense variants account for 4% of adult FSGS. In our present study, our aim is to determine whether PEC-mediated podocyte regeneration is impaired in mice with a pathogenic Pax2 missense variant (Pax2-MV). Method FSGS was induced in wildtype and Pax2-MV mice using Adriamycin. Urine samples were collected from mice every week for 28 days to measure proteinuria. At 28 days after Adriamycin injury, kidneys were collected for histopathology, transmission electron microscopy, and immunofluorescence staining for PAX2 and podocyte marker WT-1. Glomeruli from Adriamycin-injured mice were also isolated through intra-cardiac perfusion with Dynabeads and subjected to mass-spectrometry for proteomics analysis. Proteins with statistically-significant fold changes in expression levels were further analyzed by Gene Ontology to determine biological and molecular processes involved in pathogenesis. Kidney tissue from an FSGS patient with a pathogenic PAX2 missense variant was also examined by histopathology and PAX2 immunohistochemistry. Results Compared to wildtype, Pax2-MV mice were more susceptible to worsened FSGS after Adriamycin, as demonstrated by worsened glomerular scarring, increased podocyte injury and loss, and increased proteinuria, associated with increased morality in Pax2-MV mice (Fig. 1A-B). Wildtype mice showed fewer PAX2-expressing PECs after injury, accompanied by the occurrence of PAX2/WT-1 co-expressing glomerular tuft cells (Fig. 2A-B). These suggest that in Adriamycin-injured wildtype, PAX2-expressing PECs move to the glomerular tuft, where they trans-differentiate to a podocyte fate. In contrast, Pax2-MV mice showed no changes in numbers of PAX2-expressing PECs after injury, associated with fewer PAX2/WT-1 co-expressing glomerular tuft cells compared to injured wildtype (Fig. 2A-B). These suggest that in Adriamycin-injured Pax2-MV mice, PAX2-expressing PECs are unable to move to the glomerular tuft, resulting in decreased numbers of cells undergoing podocyte trans-differentiation. A subset of ectopic PAX2-expressing glomerular tuft cells after injury was increased in Pax2-MV mice (Fig. 2A), suggesting a pathologic process given the worse outcomes observed in these mice. Consistent with findings from injured Pax2-MV mice, we found that in an FSGS patient with a pathogenic PAX2 missense variant, PAX2-expressing glomerular tuft cells overlap with areas of glomerular scarring, while in a normal human kidney biopsy, PAX2 expression is only observed in PECs but not within glomerular tuft cells (Fig. 2A). Lastly, proteomics Gene Ontology analysis of isolated glomeruli demonstrated biological and molecular processes indicative of maladaptive glomerular repair in injured Pax2-MV mice, including decreased regulation of cell shape, decreased cell-cell junction assembly and decreased actin and cadherin binding compared to wildtype (Fig. 2C). Conclusion Altogether, our findings suggest that PAX2-expressing PECs migrate to the glomerular tuft to assist in podocyte regeneration in wildtype. However, this mechanism is impaired in Pax2-MV mice, leading to worsened podocyte loss and injury and further exacerbation of glomerular disease (Fig. 2C). Our findings provide a further understanding of normal and impaired regeneration mechanisms in FSGS, and our identified pathogenic mechanisms can be further harnessed for future therapeutic development.

Ferroptosis and hepatocellular carcinoma: the emerging role of lncRNAs

Hepatocellular carcinoma is the most common form of primary liver cancer and poses a significant challenge to the medical community because of its high mortality rate. In recent years, ferroptosis, a unique form of cell death, has garnered widespread attention. Ferroptosis, which is characterized by iron-dependent lipid peroxidation and mitochondrial alterations, is closely associated with the pathological processes of various diseases, including hepatocellular carcinoma. Long non-coding RNAs (lncRNAs), are a type of functional RNA, and play crucial regulatory roles in a variety of biological processes. In this manuscript, we review the regulatory roles of lncRNAs in the key aspects of ferroptosis, and summarize the research progress on ferroptosis-related lncRNAs in hepatocellular carcinoma.

G-Protein and G-Protein Coupled Receptors: Implications in Regulation of Immune Response

The guanine nucleotide binding proteins (G proteins) act as molecular switches of ‘on’ and ‘off’ when bound to GTP and GDP respectively while the guanine protein coupled receptors (GPCRs) are membrane bound receptors whereby extracellular substances (ligands)communicate signals from these substances to an intracellular molecule the G-proteins which in turn bind and activate or inhibit downstream effect or molecules causing cellular responses. This review is aimed at exploring the concept and mechanism of G-proteins and GPCR and their implication in immune response. The GPCR can be activated by various physiological or pathological processes cellular metabolism, hormones, neuro-transmitters, chemokines, autocrines, paracrines, endocrine and exocrine secretions which play an important role in relaying or routing signals to several intracellular pathways. The signal transduction by the extracellular activation or inhibition of the GPCR mediate metabolic enzymes, ion channels, transporters, cellular gene transcription, migration, survival, activation, differentiation and cytokine secretion of immune cells resulting in the synthesis and regulation of embryonic development, gonadal development, learning /memory organismal homeostasis, hematopoiesis and immune dynamics. Therefore, G proteins and GPCRs signaling systems are key determinants in innate and adaptive immunity. The signal transduction of G-Protein and GPCR by cytokine chemotaxis as Chronic inflammatory mediators is associated with tumorigenesis, metastasis with potential antagonism for appropriate targeted therapy.

#860 The role of immunocorrection in the treatment of patients chronic pyelonephritis

Abstract Background and Aims Chronic pyelonephritis in the structure of extragenital pathology in pregnant women occupies a leading place due to the high prevalence, complicated course of pregnancy and childbirth. Chronic pyelonephritis (CP) triggers a cascade of reactions, primarily immune, at the systemic and local levels. With HP the role of the immune system in pregnant women is not only a pathological process occurring in the kidneys, but also takes into account a number of changes in the restructuring of the immune system of a pregnant woman. In relation with the above, the study of the role of the immune system of pregnant women with chronic pyelonephritis is of great interest. To study the effectiveness of immunocorrective therapy in the complex treatment of patients with chronic pyelonephritis. Method 43 pregnant women with CP were examined, the average age was 28.3 ± 0.2 years. The study included clinical research methods (survey, questionnaire, objective examination). Of the laboratory methods a general analysis of blood, urine, urine analysis according to Nechiporenko, tank. urine culture, biochemical blood tests and instrumental (ultrasound) research methods. Cellular immunity was studied by monoclonal antibodies, humoral immunity was studied by Mancini immunodiffusion. Among the surveyed women, the first group of upp was pregnant women (n = 14) receiving traditional therapy. The second group of 13 pregnant women with CP on the background of traditional therapy received ozone therapy (basic treatment + 400 ml of saline solution ozonated to a concentration of ozone in a liquid of 4 mg/l, intravenously, drip). The third group of 15 pregnant women with CP - with traditional therapy received ozone therapy and immune-regulating therapy with polyoxidonium. Results The characterization of clinical and laboratory data on the background of therapy showed significant shifts, as lower back pain decreased by 85.71%, 92.31% and 93.33%, respectively, in both the first and second and third groups. Indicators such as dysuria decreased by 92.85%, 92% and 93% (respectively in the first, second and third groups). Leukocyturia decreased significantly on the fifth day by 71.43%, 84.62%, 86.67% (in the first, second and third groups, respectively). Results indicators of eradication of pathogens after treatment in combination with traditional therapy with ozone and immune-correcting therapy, they were significantly better than in the ozone therapy group against the background of traditional therapy and in the traditional treatment group (86.6%, 84.6% and 64.28%, respectively). As a result of the study, significant decreases in cellular immunity indicators, in particular CD3, CD4, were revealed before the treatment of patients and CD8 and revealed a significant increase in IgA, IgM. After the treatment in the treatment group in combination significant increases in the number of CD3, CD4 and CD8 were revealed by traditional therapy with ozone- and immune-regulating therapy (by 76.1%, 64.68% and 58.3%, respectively), compared with the group of women who received only ozone therapy against the background of traditional therapy and in the group of traditional treatment (66.23%, 59.54%, 54.2% and 61.12%, 54.68%, 51.4%, respectively), and normalization of immunoglobulins A was noted and M. Conclusion Along with the improvement of clinical and laboratory data in CP normalization of cellular and humoral immunity indicators was noted in pregnant women. Additional assignment to traditional therapy for chronic pyelonephritis of pregnant women, ozone therapy and therapy for correction of the immune system is a more effective treatment method.

Recent advances in potential therapeutic targets of ferroptosis‑associated pathways for the treatment of stroke (Review).

Stroke is a severe neurological disease that is associated with high rates of morbidity and mortality, and the underlying pathological processes are complex. Ferroptosis fulfills a significant role in the progression and treatment of stroke. It is well established that ferroptosis is a type of programmed cell death that is distinct from other forms or types of cell death. The process of ferroptosis involves multiple signaling pathways and regulatory mechanisms that interact with mechanisms inherent to stroke development. Inducers and inhibitors of ferroptosis have been shown to exert a role in the onset of this cell death process. Furthermore, it has been shown that interfering with ferroptosis affects the occurrence of stroke, indicating that targeting ferroptosis may offer a promising therapeutic approach for treating patients of stroke. Hence, the present review aimed to summarize the latest progress that has been made in terms of using therapeutic interventions for ferroptosis as treatment targets in cases of stroke. It provides an overview of the relevant pathways and molecular mechanisms that have been investigated in recent years, highlighting the roles of inducers and inhibitors of ferroptosis in stroke. Additionally, the intervention potential of various types of Traditional Chinese Medicine is also summarized. In conclusion, the present review provides a comprehensive overview of the potential therapeutic targets afforded by ferroptosis‑associated pathways in stroke, offering new insights into how ferroptosis may be exploited in the treatment of stroke.

Evaluation of Anterior Scleral Thickness and Angle Parameters in Eyes with Pseudoexfoliation Syndrome and Glaucoma.

The relationship between anterior scleral thickness and scleral spur length was disrupted in eyes with pseudoexfoliation, and scleral spur length was shorter in eyes with pseudoexfoliative glaucoma. To evaluate anterior scleral thickness (AST) in eyes with pseudoexfoliation (PX) and to examine the relationship between AST and Schlemm's canal (SC), trabecular meshwork (TM), and scleral spur (SS). Thirty-eight patients with PX syndrome (PXS), thirty-eight patients with pseudoexfoliative glaucoma (PXG), and thirty-eight healthy subjects were included in the study. Using sweep source anterior segment optical coherence tomography, AST (0, 1, 2, and 3mm from the SS), SC, and TM were visualized in the nasal and temporal areas, and measurements were compared between groups. The relationships between corneal thickness, TM, SS, SC, and AST were then evaluated. In all groups, the AST, SC, and TM measurements were similar (P>0.05). In the PXG group, SS lengths in the temporal area were shorter than those in the control and PXS groups (P=0.012). There were significant correlations between TM length and AST in the PXG group (P<0.05). The SS length exhibited moderately positive correlations with SC length and mean TM thickness in the PXG (P<0.05). There was a significant relationship between AST0 and SS in healthy eyes (P<0.05), but not in other eyes. The shorter SS length observed in eyes with PXG may be a sign of structural changes. Additionally, disruption of the relationship between AST and SS may be an early sign of pathological processes, especially in eyes with PXS, and may require closer follow-up of these eyes.