Cell Ratio Research Articles

Myocardia-Injected Synergistically Anti-Apoptotic and Anti-Inflammatory Poly(amino acid) Hydrogel Relieves Ischemia-Reperfusion Injury.

Reperfusion therapy is the most effective treatment for acute myocardial infarction, but its efficacy is frequently limited by ischemia-reperfusion injury (IRI). While antioxidant and anti-inflammatory therapies have shown significant potential in alleviating IRI, these strategies have not yielded satisfactory clinical outcomes. For that, a thermo-sensitive myocardial-injectable poly(amino acid) hydrogel of methoxy poly(ethylene glycol)45-poly(L-methionine20-co-L-alanine10) (mPEG45-P(Met20-co-Ala10), PMA) loaded with FTY720 (PMA/FTY720) is developed to address IRI through synergistic anti-apoptotic and anti-inflammatory effects. Upon injection into the ischemic myocardium, the PMA aqueous solution undergoes a sol-to-gel phase transition and gradually degrades in response to reactive oxygen species (ROS), releasing FTY720 on demand. PMA acts synergistically with FTY720 to inhibit cardiomyocyte apoptosis and modulate pro-inflammatory M1 macrophage polarization toward anti-inflammatory M2 macrophages by clearing ROS, thereby mitigating the inflammatory response and promoting vascular regeneration. In a rat IRI model, PMA/FTY720 reduces the apoptotic cell ratio by 81.8%, increases vascular density by 34.0%, and enhances left ventricular ejection fraction (LVEF) by 12.8%. In a rabbit IRI model, the gel-based sustained release of FTY720 enhanced LVEF by an additional 7.2% compared to individual treatment. In summary, the engineered PMA hydrogel effectively alleviates IRI through synergistic anti-apoptosis and anti-inflammation actions, offering valuable clinical potential for treating myocardial IRI.

Sustainable streptomycin-based Ag nanoparticle synthesis and study of the cytotoxicity.

Antibiotic resistance is one of the major problems of our time, which can be addressed by the use of different nanoparticles synthesized using antibiotics. The interaction between nanoparticles and biological systems is dynamic, which makes them extremely powerful for applications in the biomedical field. In this work, silver nanoparticles (Ag NPs) were synthesized with streptomycin and characterized by TEM, XRD, and UV-VIS. The synthesis resulted in the formation of spherical particles in the size range of 10-20nm, which did not show any signs of aggregation after several days, facilitating their ease of use. Cytotoxicity studies of the synthesized AgNPs were performed by flow cytometry on A549 lung cancer cells after 24h of exposure. At a concentration of 0.02mM AgNP, the live cell ratio did not differ significantly from the control, but LC50 value was between 1.7 and 1.9mM. The change in mitochondrial activity was examined after 4h of exposure, and the results showed that AgNPs synthesized with streptomycin induce a decrease in mitochondrial activity at concentrations as low as 0.01mM Ag NP. In this study, we have shown that antibiotic-streptomycin-stabilized nanoparticles play a dual role by both reducing and stabilizing silver nanoparticles without the need for any undesirable additive. Nanostructured silver particles synthesized with antibiotics were effective against the cancer cell line used. This is most probable because damage to the mitochondria induces the production of free radicals leading to severe cell damage. The research contributes to a deeper understanding of the effects of nanoparticles on cancer cells. The ability to stabilize silver nanoparticles with antibiotic-loaded nanoparticles could enhance therapeutic efficacy and open new opportunities for the design and development of nanomedicines for use in various biomedical fields such as wound healing, drug delivery, and antimicrobial coatings.

Disease Aggravation With Age in an Experimental Model of Multiple Sclerosis: Role of Immunosenescence.

The onset of multiple sclerosis (MS) in older individuals correlates with a higher risk of developing primary progressive MS, faster progression to secondary progressive MS, and increased disability accumulation. This phenomenon can be related to age-related changes in the immune system: with age, the immune system undergoes a process called immunosenescence, characterized by a decline in the function of both the innate and adaptive immune responses. This decline can lead to a decreased ability to control inflammation and repair damaged tissue. Additionally, older individuals often experience a shift toward a more pro-inflammatory state, known as inflammaging, which can exacerbate the progression of neurodegenerative diseases like MS. Therefore, age-related alterations in the immune system could be responsible for the difference in the phenotype of MS observed in older and younger patients. In this study, we investigated the effects of age on the immunopathogenesis of experimental autoimmune encephalomyelitis (EAE). Our findings indicate that EAE is more severe in aged mice due to a more inflammatory and neurodegenerative environment in the central nervous system. Age-related changes predominantly affect adaptive immunity, characterized by altered T cell ratios, a pro-inflammatory Th1 response, increased regulatory T cells, exhaustion of T cells, altered B cell antigen presentation, and reduced NK cell maturation and cytotoxicity. Transcriptomic analysis reveals that fewer pathways and transcription factors are activated with age in EAE. These findings allow us to identify potential therapeutic targets specific to elderly MS patients and work on their development in the future.

The association between hair cortisol and burnout is moderated by age, psychosocial, and immunological markers

BackgroundExhaustion and depersonalization are the core symptoms of the occupational burnout. However, burnout is not an all-or-nothing phenomenon, but can occur in a milder to moderate form in otherwise healthy employees. In the last two decades hair cortisol concentrations (HCC) were increasingly related to the cumulative effect of psychosocial stress at work. We analyzed data of the Dortmund Vital Study (Clinicaltrials.gov: NCT05155397) to explore the relationship of HCC and burnout symptoms. Moreover, we asked whether the HCC – burnout association was moderated by work ability, chronic stress, neuroticism, depressive symptoms, and stress-related immunological biomarkers such as T cell concentration, CD4/CD8 cell ratio, and proinflammatory cytokines TNF- α, IL-6, and IL-18. MethodsBurnout was assessed by the Oldenburg Burnout Inventory (OLBI), and the Maslach Burnout Inventory (MBI) in 196 working adults aged between 20 and 65 years (mean age 42.2 years). Several self-reported variables and biomarkers were collected. ResultsThe results showed an association between HCC and the burnout measures. A series of moderator analyses revealed that the association between HCC and burnout symptoms was substantial for low work ability, high chronic stress level, high neuroticism level, and mild to moderate depressive symptoms. Immunological markers moderated the HCC – burnout association for high concentrations of T cells, low CD4/CD8 ratio and low IL-6, IL-18 and TNF-α concentrations. These interactions were moderated by age showing the largest impact in middle-aged to older individuals. ConclusionsThe present findings shed light on the complex interaction between burnout symptoms and work ability, chronic stress, personality, and the endocrinological and immunological responses across the working lifespan. These parameters should be considered when assessing the risk for developing burnout and validating the diagnosis of burnout. Trial RegistrationClinicalTrials.gov NCT05155397; https://clinicaltrials.gov/ct2/show/NCT05155397

Role of intraperitoneal paclitaxel in eosinophil activation and recruitment in the peritoneal cavity and anti-tumor effects against peritoneal metastasis from gastric cancer.

471 Background: Recent studies indicate that the efficacy of chemotherapy is significantly influenced by the tumor immune microenvironment. Peritoneal cavity contains many immune cells, however, the relationship between immune response within the peritoneal cavity and tumor response remains poorly understood. Methods: Single cell suspensions were obtained from ascites or peritoneal lavages from 41 patients with PM from GC. In 28 patients, cells were collected both before and after 1-3 courses of IP chemotherapy. These samples were immunestained with mAbs targeting to specific lymphoid or myeloid subsets, and their proportions in CD45(+) leukocytes were analyzed using flowcytometry. Eosinophils were purified by magnetic cell sorting for subsequent RNA-Seq analysis. Results: Among 41 patients with PM, tumor leukocyte ratio (TLR) calculated as CD326(+) tumor cells divided by CD45(+) leukocytes varied from 0.002% to 58.1%. TLR was not correlated with any ratios of immune cells, however, the ratios of CD8(+) T cells, NK cells and CD16(-) CD193(+) eosinophils tended to decrease with the increase in TLR. Conversely, the ratios of CD19(+) B cell, CD14(+) macrophages and CD16(+) neutrophils increased with elevated TLR. Mean survival time of these patients was 17.7 months. None of the leukocyte subsets showed significant association with patient outcome. In 28 patients, the ratios of lymphocytes mostly decreased with significant difference in CD4(+) T cells and CD19(+) B cells after IP treatment. In contrast, the ratio of CD11b(+) myeloid cells increased in peritoneal cavity after IP treatment. Among the myeloid cells, the ratio of CD16(-) CD193(+) eosinophils significantly increased. The change was particularly prominent in in post-treatment negative peritoneal lavage cytology (CY0) patients (M=0.47% vs M=10.0%, p<0.0001), while the difference was not significant in post-treatment positive peritoneal lavage cytology (CY1) patients (M=0.93% vs M=0.97%, p=0.84). Patients with eosinophil ratio of ≥2% after IP chemotherapy in peritoneal cavity had significantly longer overall survival compared to those with lower eosinophil ratios (17.3 mo vs 26.7 mo, p=0.034; HR =0.23[0.06-0.89]). The peritoneal eosinophils after IP treatment exhibited elevated levels of CD11b and CD63 expression compared to circulating eosinophils (p<0.01), while SSC-A levels tended to be lower. Gene ontology pathways revealed enrichment of cytokine-mediated signaling pathway, extracellular matrix organization and response to interferon-gamma in peritoneal eosinophils (p<0.0001). Conclusions: IP-PTX induces eosinophil activation and recruitment in the peritoneal cavity, while also affecting other types of immune cells. These effects may potentially contribute to the anti-tumor response against peritoneal metastases.

Blood Cell Ratio Combinations for Diagnosing Periprosthetic Joint Infections: A Preliminary Study.

Periprosthetic joint infection (PJI) is a serious complication following total joint arthroplasty (TJA), which requires prompt and accurate diagnosis for effective management. Many biomarkers have been used for PJI diagnosis; however, the identification of the most effective inflammatory biomarker combination for optimal diagnostic accuracy may be poorly reported. In this prospective, multi-center study, a total of 269 individuals undergoing knee or hip revision arthroplasty were recruited and subsequently categorized based on 2018 ICM PJI criteria into two groups: 93 with periprosthetic joint infection (PJI) and 176 with aseptic failure (AF). Various preoperative biomarkers were analyzed and compared, including C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), neutrophil-to-lymphocyte ratio (NLR), CRP-to-albumin ratio (CAR), CRP-Albumin-lymphocyte ratio (CALLR), platelet-to-lymphocyte ratio (PLR), platelet-to-albumin ratio (PAR), and neutrophil-to-albumin ratio (NAR). The diagnostic performance of these biomarkers was evaluated using ROC curve analysis and the area under the curve (AUC). Additionally, the Youden index was used to determine optimal threshold values, and positive predictive value (PPV) and negative predictive value (NPV) were calculated to evaluate diagnostic precision. In the PJI group, levels of PAR, CAR, and CALLY were notably higher compared to the AF group, reaching statistical significance (P < 0.05). PAR and CAR were confirmed to have high diagnostic values, with AUC values of 0.779 and 0.718, respectively. CALLY exhibited moderate diagnostic effectiveness, with an AUC of 0.647. When PAR was combined with CRP and ESR, sensitivity and specificity notably improved to 93.8% and 92.5%, respectively. However, subgroup analysis revealed no significant differences in combined inflammatory biomarker levels between the two groups. PAR and CAR prove to be effective combined inflammatory biomarkers for PJI diagnosis, whereas other markers exhibited limited diagnostic utility for PJI.

Platelet Volume Parameters as Predictors of Valvular Thrombosis Risk in Patients with Aortic and Mitral Valve Replacement.

Percutaneous valve implantation or surgical replacement with mechanical or biological valves are standard therapies for severe valvular heart diseases. Prosthetic valve thrombosis, though rare, is a serious complication, particularly with mechanical prostheses. This study aimed to investigate the predictive value of platelet volume parameters, including mean platelet volume (MPV), platelet distribution width (PDW), and platelet-large cell ratio (P-LCR), for valvular thrombosis risk in patients undergoing valve replacement therapy. A retrospective cross-sectional study was conducted from May 2002 to May 2020, involving 108 patients with a history of mitral or aortic valve replacement and valvular thrombosis, and 216 controls with a history of valve surgery without valvular malfunction. PDW was significantly associated with an increased risk of thrombosis after adjusting for confounders, while MPV showed a clinical difference but did not reach statistical significance. P-LCR did not exhibit a significant association. These findings suggest PDW as a potential predictor of valvular thrombosis in such patients. The ease of measuring platelet volume parameters suggests their utility in routine hematological analysis for identifying patients at higher risk of valvular thrombosis post-replacement surgery. Further studies are warranted to validate these findings and explore additional laboratory markers, such inflammatory markers, for thrombotic risk assessment in this population.

EXPRESS: Unlocking the Immune Puzzle: T Cell exhaustion in cirrhosis and implication for immunotherapy.

Cirrhosis, an advanced stage of liver diseases, induces Cirrhosis-Associated Immune Dysfunction Syndrome (CAIDS) characterized by both innate and adaptive immune dysfunction. Inflammation triggered by factors such as alcohol, viruses, toxins, and cholesterol induce metabolic reprogramming of both innate and adaptive immune cells. Our study specifically sought to investigate the compromised adaptive immune response in cirrhosis by focusing on assessing T-cell exhaustion and activation markers on helper and cytotoxic T cells. A prospective observational study involving 19 liver cirrhosis patients and 36 healthy controls was conducted. Hepatic decompensation degree was assessed using various parameters including serum bilirubin, albumin, international normalized ratio, ascites and hepatic encephalopathy. T cell activation (CD38, CD44, CD69, HLADR), and exhaustion markers (CTLA-4, PD-1, TIM-3, LAG-3) were assessed on helper and cytotoxic T cells by flow cytometry. Cirrhosis patients showed reduced T cells with no alteration in CD4:CD8 T cell ratio. Among activation markers, HLADR showed increased expression on CD8+ T cells (P=0.031). Regarding exhaustion markers LAG-3 and TIM-3 exhibited increased expression in cirrhotic patients compared to controls in both CD4 and CD8 T cells (P=0.004, P=0.016, P=0.001, P=0.004, respectively). Neither cirrhotic nor healthy controls showed CTLA expression. PD-1 did not differ significantly between the two groups. Co-expression of PD-1/TIM-3 on CD8+ T cells was notably higher in cirrhotic patients (P<0.002). The observation of impaired adaptive immunity with notable T cell exhaustion and activation in cirrhosis underscores the potential relevance of immunotherapy.

Neuregulin1 ameliorates metabolic dysfunction-associated fatty liver disease via the ERK/SIRT1 signaling pathways.

Neuregulin (NRG) family is involved in energy metabolism, among which NRG1 is a neuregulin proved to play a protective role in MAFLD cells. But the presice echanism has not been fully illustrated. This study aimed to investigate the role of NRG1 via the ERK/SIRT1 signaling in the pathogenesis of MAFLD. C57BL/6 mice were fed with high-fat diet for 8 weeks, and then injected with NRG1 (0.3mg/kg/d) and PD98059 (0.3mg/kg/d) via tail vein for 5 weeks. HepG2 cells induced by oleic acid and palmitic acid were treated with 20ng/mL NRG1 and 10µmol/L PD98059. The changes of histopathological, biochemical indexes, inflammatory factors, lipid metabolism, apoptosis and autophagy parameters were measured. The expressions of NRG1 in MAFLD cell and animal models were significantly lower than that in the control group. After the intervention of ERK inhibitor PD98059, the expression of NRG1 decreased significantly in vivo, but no significant change was observed in vitro. Moreover, NRG1 ameliorated hepatic steatosis, enhanced cell viability, reduced cell apoptosis, and attenuated liver injury both in vitro and in vivo. After NRG1 intervention, the expressions of ERBB2, ERBB3, p-ERK1/2, SIRT1 and p-FOXO1 as well as the LC3II/I ratio in MAFLD cells and liver tissues of MAFLD mice were significantly increased, while the expression of SREBP1c was decreased. The aforementioned therapeutic effect of NRG1 was lost after the intervention of PD98059. NRG1 might play a protective role in the pathogenesis of MAFLD by activating the downstream ERK1/2 through ErbB2-ErbB3, which promotes the expression of SIRT1 and autophagy markers. This study might indicate a new therapeutic strategy for MAFLD.

The Potential Mechanism of Cuproptosis in Hemocytes of the Pacific Oyster Crassostrea gigas upon Elesclomol Treatment

Cuproptosis is a novel cell death dependent on mitochondrial respiration and regulated by copper. While the study of it is mainly focused on tumor therapy, in the present study, two key cuproptosis-related genes, ferredoxin (FDX1) and dihydrolipoamide S-acetyltransferase (DLAT) homologs (designated as CgFDX1 and CgDLAT), were identified from Crassostrea gigas. CgFDX1 has a Fer2 domain with a 2Fe-2S cluster forming a unique ferredoxin. CgDLAT is composed of a biotin_lipoyl domain, an E3-binding domain, and a 2-oxoacid_dh domain. CgFDX1 and CgDLAT mRNA were expressed in all the examined tissues. After elesclomol treatment, both mRNA and protein expressions of them were reduced in the hemocytes. The mortality rate of the hemocytes increased significantly, and the hemocytes were accompanied with noticeable adhesive abnormalities and heightened secretion after elesclomol treatment. Additionally, the accumulation or depletion of actin was observed in the hemocytes. The integrity of the double membrane structure of the mitochondria was compromised, and the organization of mitochondrial cristae was disrupted. The contents of copper, malondialdehyde (MDA), pyruvic acid and mitoSOX as well as the ratio of cells with low mitochondrial potential increased significantly in the hemocytes upon elesclomol treatment and the content of citric acid decreased significantly. These findings suggest the potential presence of cuproptosis in oysters and its activation mechanism is relatively conserved in evolution.

Mortality and immunological indicators of men who have sex with men living with HIV on antiretroviral therapy: a 10-year retrospective cohort study in Southern China

BackgroundThe proportion of people living with HIV (PLHIV) in Guangxi who are men who have sex with men (MSM) increased rapidly to nearly 10% in 2023; notably, over 95% of this particular population is currently receiving antiretroviral therapy (ART). This study aimed to describe the survival of MSM PLHIV, depict the characteristics and trends of changes in CD4+ T cell counts, CD4+/CD8+ T cell ratio, and viral load, and explore immunological indicators that may be related to mortality during different stages of treatment.MethodsImmunological indicators of MSM PLHIV receiving ART were extracted and categorized into baseline, mid-treatment, and last values. These were then incorporated into the Cox models in the form of repeated measurements to evaluate the associations (Adjusted-hazard ratios, aHRs) and 95% confidence intervals (95% CIs) of these immunological indicators with mortality at different stages.ResultsA total of 5,642 patients who met the criteria were enrolled in the study, and 160 died, representing a mortality of 2.8%. The first, median, and last values of the CD4+ count and the CD4+/CD8+ ratio in surviving patients were significantly greater than the corresponding values in nonsurviving patients (P < 0.001). Except for the first viral load (P = 0.379), the median and last viral loads of the former were significantly lower than those of the latter (P < 0.001). In Cox model, with 2,144 cases, immunological indicators of increased mortality risk included a baseline CD4+ count below 200 cells/µL (aHR: 4.58, 95% CI: 2.28–9.19), a median (aHR: 8.46, 95% CI: 3.05–23.46), and a final (aHR: 4.43, 95% CI: 1.06–18.45) ratio below 0.7 and a median (aHR: 9.47, 95% CI: 4.02–22.35) and final (aHR: 14.46, 95% CI: 4.50–46.50) viral load 100,000 copies/mL and above.ConclusionsMortality among MSM PLHIV on ART in Guangxi is relatively low, and both high viral loads and low CD4+/CD8+ T cell ratios during treatment and at recent follow-up are strongly predictive of a serious prognosis and should be closely monitored.

Platelet-white cell ratio is more strongly associated with mortality than other common risk ratios derived from complete blood counts

Complete blood count indices and their ratios are associated with adverse clinical outcomes for many acute illnesses, but the mechanisms generating these associations are not fully understood. Recent identification of a consistent pattern of white blood cell and platelet count co-regulation during acute inflammatory recovery provides a potentially unifying explanation. Here we show that the platelet-to-white-cell ratio, which was selected based on this conserved recovery pattern, is more strongly associated with mortality than other blood count markers and ratios in four important illnesses involving acute inflammation: COVID-19, acute heart failure, myocardial infarction, and stroke. Patients recovering well from these acute illnesses tend to follow a joint white cell and platelet trajectory that can be reduced to this one-dimensional ratio. The platelet-to-white-cell ratio’s association with prognosis is consistent with recently identified inflammatory dynamics and may provide a convenient and interpretable summary of patient inflammatory state.

Abstract P019: Spatiometabolic mapping of cytoarchitectural changes in lung adenocarcinoma following radiotherapy

Abstract Understanding how radiotherapy induces metabolic reprogramming and reshapes the cytoarchitecture of the tumor microenvironment (TME) is essential for improving treatment outcomes and overcoming resistance. Despite its importance, no studies have used computational tools to quantitatively assess these cytoarchitectural rearrangements in the TME following radiotherapy. In previous work, we developed a quantitative method called “colocatome analysis,” a spatial -omic approach that catalogs statistically significant colocalizations between pairs of cell subpopulations or states. Through a normalization process, this method enables direct comparisons of spatial features across different conditions, including comparisons between in vitro models and tissue samples. We applied colocatome analysis to multiplexed immunofluorescence images of lung adenocarcinoma specimens, examining differences between naïve and recurrent tumors following radiotherapy using a panel of markers that represent primary tumor compartments (endothelial, malignant, fibroblast, and immune) and include markers for the hexosamine biosynthesis pathway (HBP), glycans, key metabolic enzymes, proliferation, cell death, DNA damage, hypoxia, and EMT. Our preliminary results indicate significant cytoarchitectural differences: tumors that recurred after radiotherapy exhibited a lower epithelial-to-mesenchymal cancer cell ratio and were enriched in FAP+ cancer-associated fibroblasts (CAFs). Although the overall proportions of macrophages and other immune cells remained consistent across samples, their spatial organization differed. In naïve tumors, immune cells were largely segregated from cancer cells, while recurrent tumors showed a more homogeneous mixture of cancer and immune cells. Additionally, CAFs were markedly increased in recurrent tumors. Lastly, cancer cells in naïve tumors displayed high levels of O-GlcNAc, which significantly decreased in recurrent tumors. These findings provide insights into the impact of metabolic reprogramming towards HBP in the TME and highlight its potential role in mediating radioresistance. Citation Format: Gina Bouchard.Spatiometabolic mapping of cytoarchitectural changes in lung adenocarcinoma following radiotherapy.[abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Targeted Therapies in Combination with Radiotherapy; 2025 Jan 26-29; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(2_Suppl):Abstract nr P019

Preoperative diagnostic efficacy of novel blood markers white blood cell ratio and fibrinogen levels in periprosthetic joint infection

To investigate the clinical utility of novel of new hematological markers in the preoperative diagnosis of periprosthetic joint infection (PJI). A retrospective analysis was conducted on a total of 149 patients who underwent revision of total hip arthroplasty (THA) or total knee arthroplasty (TKA) at a single center between January 2016 and June 2022, including 63 males and 86 females, aged from 47 to 93 years old with an average of (69.5±11.8) years old. Of them, 46 were diagnosed as PJI(PJI group), including 22 males and 24 females. The mean age was (71.3±12.5) years old. The body mass index (BMI) was (26.4±3.1) kg·m-2. And 103 patients were diagnosed as aseptic prosthesis loosening (aseptic group), including 41 males and 62 females. The mean age was (68.7±11.4) years old. The BMI was (25.8±3.5) kg·m-2. Preoperatively analyzed clinical parameters included C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), albumin, globulin, albumin-to-globulin ratio (AGR), plasma D-dimer, and plasma fibrinogen. The receiver operating characteristic curve (ROC), sensitivity, and specificity analysis were employed to compare the diagnostic value of each blood marker in preoperative PJI diagnosis. In the PJI group, the levels of CRP were 16.6 (7.6, 4.5) mg·L-1, ESR was 17.0 (12.8, 35.5) mm·h-1, plasma D-dimer was 1.0 (0.5, 3.1) μg·L-1, and plasma fibrinogen was 4.2 (3.2, 3.1) mg·L-1;all of which were higher compared to the aseptic group with CRP at 4.2 (2.6, 7.8) mg·L-1, ESR at 12.0(8.0, 20.0 )mm·h-l, D-dimer at 0.4(0.2, 0.7)μg·L-1, and fibrinogen at 2.8(2.4, 3.3 ) g·L-1(P<0.05). However, the albumin level of 35.3 (32.3, 37.5) g·L-1 and the WBC ratio of 1.0(0.9, 1.1) in the PJI group were significantly lower compared to the aseptic group with levels of 39.8 (36.1, 41.8) g·L-1 and 1.4 (1.3, 1.5), respectively (P<0.05). Only the area under the curve (AUC) of AGR and plasma fibrinogen were greater than 0.8. The optimal predictive cut-1off, AUC, sensitivity and specificity were 3.4 g·L-1, 0.820, 69.57% and 84.47% for plasma fibrinogen; 1.18, 0.813, 82.61% and 78.64% for AGR, respectively. AGR and plasma fibrinogen are promising blood markers for improving the diagnosis of PJI.

T-cell immune status in patients with acute exacerbation of chronic obstructive pulmonary disease: a case-control study

IntroductionImmune inflammatory response plays an important role in chronic obstructive pulmonary disease (COPD). However, the cellular immune status of patients with COPD at different phases is unclear. Herein, we aim to investigate the distribution and functional status of T cell subsets in different phases of COPD (acute exacerbation of COPD [AECOPD] and stable COPD [SCOPD]).MethodsThis is an observational case-control study undertaken in West China Hospital. The distribution of T cell subsets in peripheral blood of AECOPD, SCOPD, and healthy controls (HCs) was measured using multi-color flow cytometry, and the functional status was analyzed by additional staining of activation markers.ResultsA total of 43 HCs, 43 SCOPD patients, and 64 AECOPD patients were evaluated. The total number and percentage of lymphocytes and the CD4+/CD8+ T cells ratio were significantly lower in AECOPD patients when compared to HCs. HLA-DR expression in CD3+, CD4+, CD8+, CD8+ TCR aβ, and CD4+ TCR aβ T cells was upregulated in the AECOPD group. Similarly, the expressions of HLA-DR, CD57, and PD-1 were higher in T cell subsets in the AECOPD group. Compared with the SCOPD and HC groups, the AECOPD had a significantly lower proportion of CD4+CD27+CD28+ T cells, but opposite results were found for CD4+CD27-CD28- T cells. In addition, the proportion of CD4+CD39+ T cells and CD4+CD25+FoxP3+ T cells was significantly higher in the AECOPD and SCOPD groups when compared to the HC group (P &amp;lt; 0.05).DiscussionThe distribution of nearly half the T cell subsets in AECOPD patients was significantly different from that in SCOPD patients and HCs. AECOPD patients may have cellular immune suppression, immune dysfunction, abnormal activation, and higher senescence depletion of T cells.

Cell viability assessment by using GelRed/SYTO 9-based double staining

Cell viability assessment plays a crucial role in biological research, pharmaceutical development, and toxicological identification. Here, we used GelRed, a sensitive and safer nucleic acid dye, to selectively label dead cells with red fluorescence (FL) thus distinguishing dead cells from live ones. Further more, the combined use of GelRed and SYTO 9 (another nucleic acid dye) enabled the clear differentiation in FL spectra between the two physiological statuses. The GelRed and SYTO 9 concentrations were optimized to obtain the highest FL ratio of dead to live cells. The GelRed/SYTO 9-based double staining could quantify the cell viability through flow cytometry analysis, with a good correlation between the detected and theoretical dead cell ratios. Compared with traditional prodium iodide (PI) staining, the GelRed/SYTO 9-based double staining showed high accuracy in quantifying dead cell of low levels. The as-developed staining method could be used in biomedical research to accurately measure the cytotoxic effect of various substances in living cells.

Trajectories of platelet indices and their association with mortality in the ICU—a longitudinal cohort study

While thrombocytopenia’s link to mortality is known, the prognostic impact of longitudinal trajectories of platelet indices in combination with analysis of thrombocytopenia’s mediating role remains unexplored. This is the first study that addresses this significant gap by investigating the association between seven platelet indices trajectory subphenotypes and ICU mortality, considering thrombocytopenia’s mediating influence. Four hundred and twenty-one adult ICU patients were enrolled in this longitudinal cohort study. Three trajectories were identified for each platelet index, namely: descending, stable, and ascending, and using a regression, receiver-operating characteristic curve, and mediation analysis, their associations with 90-day mortality were evaluated with the mediating effect of thrombocytopenia. The findings were adjusted (prefixed ‘a’) for covariates. The heterogeneous trajectories significantly associated with 90-day mortality included: descending platelet count (PC) [aOR, 2.75 (CI, 1.56–4.85), p = 0.0005, aAUC, 0.783], descending plateletcrit (PCT) [aOR, 3.49 (CI, 1.88–6.46), p = 0.0001, aAUC, 0.802], ascending platelet distribution width (PDW) [aOR, 2.04 (CI, 1.13–3.71), p = 0.0188, aAUC, 0.776], and ascending percent-immature platelet fraction (%-IPF) [aOR, 2.25 (CI, 1.29–3.94), p = 0.0045, aAUC, 0.778], with 11.6% (p = 0.027), 12.0% (p = 0.019), 22.1% (p = 0.011), and 15.9% (p = 0.024) effects mediated by thrombocytopenia, respectively. In contrast, ascending mean platelet volume (MPV) was significantly and independently associated with mortality [aOR, 3.04 (CI, 1.45–6.39), p = 0.0033, aAUC, 0.781], without the effect mediated by thrombocytopenia (p = 0.056). The trajectories of platelet-large cell ratio (P-LCR) and absolute-immature platelet count (A-IPF) were not significantly associated with the risk of mortality (p > 0.05). This study demonstrated that descending PC and PCT and ascending PDW and %-IPF, mediated by thrombocytopenia, and ascending MPV, without mediation by thrombocytopenia, are useful longitudinal trajectories for predicting 90-day mortality in the ICU.

In Vitro Evaluation of the Safety and Efficacy of Cibisatamab Using Adult Stem Cell-Derived Organoids and Colorectal Cancer Spheroids.

Objectives: Developing ex vivo models that replicate immune-tumor interactions with high fidelity is essential for advancing immunotherapy research, as traditional two-dimensional in vitro systems often lack the complexity required to fully represent these interactions. Methods: In this study, we establish a comprehensive 3D redirect lysis (3D-RDL) assay using colorectal cancer spheroids and adult stem cell-derived, healthy human organoids to evaluate the efficacy and safety profile of Cibisatamab, a bispecific antibody targeting carcinoembryonic antigens (CEAs) on cancer cells and CD3 on T cells. This model allows us to assess cytotoxic activity and immune responses, capturing variations in therapeutic response not observable in simpler systems. Our model integrates live imaging and cytotoxicity analyses to enable precise, real-time tracking of antibody effects on CEA-expressing tumor cells compared to healthy cells. Additionally, by standardizing effector-to-target cell ratios in each co-culture, we establish a reproducible workflow that enhances data accuracy and comparability across assays. Flow cytometry and Granzyme B release profiling further allow us to characterize immune cell activation, revealing distinct T cell activation markers and Granzyme B release patterns tied to Cibisatamab treatment. Results: Our results show that Cibisatamab effectively induces cell death in cancer spheroids with high CEA expression while being dose-dependent on target, off-tumor binding and killing on non-cancerous cells of healthy organoids with intermediate CEA levels. This highlights our model's potential to predict clinical immunotherapy outcomes, capturing complex responses like immune activation, therapeutic selectivity, and potential resistance mechanisms. Conclusions: These findings underscore the utility of our model as a reliable, physiologically relevant tool for screening new immunotherapies and advancing our understanding of tumor-immune dynamics.

Loss of Sirtuin 7 impairs cell motility and proliferation and enhances S-phase cell arrest after 5-fluorouracil treatment in head and neck cancer

Sirtuin 7 (SIRT7), a member of the sirtuin family of NAD+-dependent deacetylases, plays a vital role in cancer, exhibiting context-dependent functions across various malignancies. Our study investigates the role of SIRT7 depletion in head and neck squamous cell carcinoma (HNSCC) progression. In vitro and 3D organotypic models demonstrated that SIRT7 knock-out attenuates cancer cell viability, proliferation, and motility as well as induces downregulation of migration- and epithelial-mesenchymal transition (EMT)-related gene expression. Moreover, the SIRT7 loss results in slower organoid formation and less invasive organoid morphology, validated by vimentin downregulation. The SIRT7 loss potentiates S-phase arrest in cell cycle progression after 5-FU treatment and elevates the ratio of dead cells. Additionally, SIRT7 deletion reduces the expression of G1 phase-associated proteins, Cyclin D and CDK4. Altogether, our study highlights SIRT7 as a promising therapeutic target in HNSCC, enhancing the effectiveness of treatment modalities such as combinational treatment.

Interleukin-17A facilitates tumor progression via upregulating programmed death ligand-1 expression in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is an inflammation-associated tumor with a dismal prognosis. Immunotherapy has become an important treatment strategy for HCC, as immunity is closely related to inflammation in the tumor microenvironment. Inflammation regulates the expression of programmed death ligand-1 (PD-L1) in the immunosuppressive tumor microenvironment and affects immunotherapy efficacy. Interleukin-17A (IL-17A) is involved in the remodeling of the tumor microenvironment and plays a protumor or antitumor role in different tumors. We hypothesized that IL-17A participates in tumor progression by affecting the level of immune checkpoint molecules in HCC. To investigate the effect and mechanism of action of IL-17A on PD-L1 expression and to identify attractive candidates for the treatment of HCC. The upregulation of PD-L1 expression in HCC cells by IL-17A was assessed by reverse transcription PCR, western blotting, and flow cytometry. Mechanistic studies were conducted with gene knockout models and pathway inhibitors. The function of IL-17A in immune evasion was explored through coculture of T cells and HCC cells. The effects of IL-17A on the malignant biological behaviors of HCC cells were evaluated in vitro, and the antitumor effects of an IL-17A inhibitor and its synergistic effects with a PD-L1 inhibitor were studied in vivo. IL-17A upregulated PD-L1 expression in HCC cells in a dose-dependent manner, whereas IL-17A receptor knockout or treatment with a small mothers against decapentaplegic 2 inhibitor diminished the PD-L1 expression induced by IL-17A. IL-17A enhanced the survival of HCC cells in the coculture system. IL-17A increased the viability, G2/M ratio, and migration of HCC cells and decreased the apoptotic index. Cyclin D1, VEGF, MMP9, and Bcl-1 expression increased after IL-17A treatment, whereas BAX expression decreased. The combination of IL-17A and PD-L1 inhibitors showed synergistic antitumor efficacy and increased cluster of differentiation 8 + T lymphocyte infiltration in an HCC mouse model. IL-17A upregulates PD-L1 expression via the IL-17A receptor/phosphorylation-small mothers against decapentaplegic 2 signaling pathway in HCC cells. Blocking IL-17A enhances the therapeutic efficacy of PD-L1 antibodies in HCC in vivo.