Cell Ratio Research Articles

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.

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.

Artificial dural regeneration matrix as a substitute for autologous tissue in indirect bypass in Moyamoya disease: Investigation of a rat model of chronic cerebral hypoperfusion

Indirect bypass using autologous tissue is effective in Moyamoya disease, especially among pediatric patients. This study aimed to evaluate the effectiveness of indirect bypass using DuraGen (absorbable artificial dura mater composed of collagen matrix), as a substitute for autologous tissue in a rat model of chronic cerebral hypoperfusion. Male Wistar rats were subjected to bilateral internal carotid artery occlusion and divided into three groups: a control group without bypass surgery, a group wherein indirect bypass was performed using the temporalis muscle (encephalo-myo-synangiosis [EMS] group), and a group wherein DuraGen was used (Dura group). The ratio of the number of vascular endothelial cells, detected by antibodies to CD31 and glucose transporter type 1 (Glut-1), on the operative side to that on the non-operative side was measured and compared between the three groups. The ratio of CD31-positive cells was 1.50 ± 0.13 and 1.92 ± 0.29 in the EMS and Dura groups, respectively, and that of Glut-1-positive cells was 1.32 ± 0.10 and 1.53 ± 0.18 in the EMS and Dura groups, respectively. No significant difference was observed in the ratio of vascular endothelial cells on the bypass side between the EMS and Dura groups. Indirect bypass with DuraGen resulted in an increased ratio of vascular endothelial cells, equivalent to that of an indirect bypass with the temporalis muscle in a rat model. Thus, in an actual indirect bypass for patients with Moyamoya disease, the use of DuraGen may produce the same angiogenesis as using autologous tissue.

ADARp110 promotes hepatocellular carcinoma progression via stabilization of CD24 mRNA

ADAR is highly expressed and correlated with poor prognosis in hepatocellular carcinoma (HCC), yet the role of its constitutive isoform ADARp110 in tumorigenesis remains elusive. We investigated the role of ADARp110 in HCC and underlying mechanisms using clinical samples, a hepatocyte-specific Adarp110 knock-in mouse model, and engineered cell lines. ADARp110 is overexpressed and associated with poor survival in both human and mouse HCC. It creates an immunosuppressive microenvironment by inhibiting total immune cells, particularly cytotoxic GZMB+CD8+ T cells infiltration, while augmenting Treg cells, MDSCs, and exhausted CD8+ T cells ratios. Mechanistically, ADARp110 interacts with SNRPD3 and RNPS1 to stabilize CD24 mRNA by inhibiting STAU1-mediated mRNA decay. CD24 protects HCC cells from two indispensable mechanisms: macrophage phagocytosis and oxidative stress. Genetic knockdown or monoclonal antibody treatment of CD24 inhibits ADARp110-overexpressing tumor growth. Our findings unveil different mechanisms for ADARp110 modulation of tumor immune microenvironment and identify CD24 as a promising therapeutic target for HCCs.

The prognostic value of systemic immune-inflammation index in patients with unresectable hepatocellular carcinoma treated with immune-based therapy

BackgroundPredicting the efficacy of immune-based therapy in patients with unresectable hepatocellular carcinoma (HCC) remains a clinical challenge. This study aims to evaluate the prognostic value of the systemic immune-inflammation index (SII) in forecasting treatment response and survival outcomes for HCC patients undergoing immune-based therapy.MethodsWe analyzed a cohort of 268 HCC patients treated with immune-based therapy from January 2019 to March 2023. A training cohort of 93 patients received atezolizumab plus bevacizumab (T + A), while a validation cohort of 175 patients underwent treatment with tyrosine kinase inhibitors (TKIs) combined with anti-PD-(L)1 therapy. The SII cutoff value, determined using X-tile analysis based on overall survival (OS) in the training cohort, divided patients into high (> 752*109) and low (≤ 752*109) SII groups. Prognostic factors were identified through univariate and multivariate logistic and Cox regression analyses, and survival outcomes were assessed using Kaplan–Meier methods. The predictive accuracy of SII was evaluated using receiver operating characteristic (ROC) curves.ResultsAn optimal SII cutoff of 752*109 stratified patients into high and low SII groups. Univariate and multivariate logistic regression indicated that SII was a significant predictor of the objective response rate (ORR), which was markedly different between the low and high SII subgroups (34.72% vs. 9.52%, P = 0.019). This finding was consistent in the validation cohort (34.09% vs. 16.28%, P = 0.026). SII also demonstrated prognostic value in Cox regression and Kaplan–Meier analyses. ROC curves confirmed that SII had superior predictive accuracy compared to common clinical indicators, with predictive relevance even in AFP-negative patients. Furthermore, a lower SII was associated with a higher T cell ratio and an increased number of CD8+ T cells and Granzyme B+ CD8+ T cells in peripheral blood.ConclusionSII is a promising predictor of both therapeutic efficacy and prognosis in HCC patients undergoing immune-based treatments. Its application may enhance clinical decision-making, thereby improving patient outcomes from immune-based therapy.

Investigation of the Effects of Polyunsaturated Fatty Acid Ratios in Human SH-SY5Y Cells by in vitro Methods

Investigation of the Effects of Polyunsaturated Fatty Acid Ratios in Human SH-SY5Y Cells by in vitro Methods

Early Initiation of Bundle Sheath Cells During Leaf Development as Visualised by SCARECROW Expression in Dicotyledonous C4 Plants.

The C4 type of dicotyledonous plants exhibit a higher density of reticulate veins than the C3 type, with a nearly 1:1 ratio of mesophyll cells (MCs) to bundle sheath cells (BSCs). To understand how this C4-type cell pattern is formed, we identified two SCARECROW (SCR) genes in C4 Flaveria bidentis, FbSCR1 and FbSCR2, that fully or partially complement the endodermal cell layer-defective phenotype of Arabidopsis scr mutant. We then created FbSCRs promoter β-glucuronidase reporter (GUS) lines of F. bidentis, which showed GUS expression in BSCs and their progenitor cells. The GUS expression pattern in F. bidentis transformants and comparison with the closely related C3-type Flaveria pringlei revealed that higher-order veins were initiated in the early leaf developmental stage. Treatment with an auxin polarity transport inhibitor decreased the MC area and led to vein formation without free ends, resulting in the formation of BSCs in positions adjacent to other BSCs. However, BSC differentiation was not affected, as evidenced by BSC specific FbSCR1 expression and RuBisCO accumulation. These results indicate that polar auxin transport is important for MC proliferation and/or differentiation, which leads to the formation of a C4-type cell pattern in which MCs and BSCs are equally adjacent.

MicroRNA-155 Inhibition Activates Wnt/β-catenin Signaling to Restore Th17/Treg Balance and Protect against Acute Ischemic Stroke.

Acute ischemic stroke (AIS) is a dangerous neurological disease associated with an imbalance in Th17/Treg cells and abnormal activation of the Wnt/β-catenin signaling pathway. This study aims to investigate whether inhibition of miR-155 can activate the Wnt/β-catenin signaling pathway to improve Th17/Treg imbalance and provide neuroprotective effects against stroke. We employed a multi-level experimental design. Firstly, we analyzed the differential gene expression between the miR-155 antagomir-treated group and the control group using high-throughput sequencing to identify potential target genes. Subsequently, we conducted functional and pathway enrichment analysis of the differentially expressed genes using bioinformatics tools. Next, we performed in vivo animal experiments using a mouse model to validate the impact of miR-155 antagomir treatment on the Wnt/β-catenin signaling pathway and improvement of Th17/Treg cell ratios. Lastly, we conducted in vitro cell experiments to validate our findings further. High-throughput sequencing results showed significant differential expression between the miR-155 antagomir-treated group and the control group (BioProject: PRJNA1152758, SRA IDs: SRR30410532, SRR30410531, SRR30410530 for the disease group; SRR30410529, SRR30410528, SRR30410527 for the control group). Bioinformatics analysis revealed potential target genes associated with the Wnt/β-catenin signaling pathway and Th17/Treg cell imbalance. In vitro experiments demonstrated that miR-155 antagomir treatment significantly activated the Wnt/β-catenin signaling pathway and improved Th17/Treg cell ratios. In vivo, animal experiment results indicated that miR-155 antagomir treatment exhibited significant neuroprotective effects against AIS. This study demonstrates that miR-155 antagomir can improve Th17/Treg cell imbalance by activating the Wnt/β-catenin signaling pathway and exhibiting neuroprotective effects against AIS in a mouse model. These findings provide crucial support for miR-155 as a potential therapeutic strategy for stroke and lay the foundation for further research.Significance Statement This study identifies miR-155 as a pivotal regulator of the Th17/Treg cell balance and Wnt/β-catenin signaling pathway in AIS. By inhibiting miR-155, we demonstrate the potential to enhance neuroprotection and modulate immune responses, offering a promising therapeutic avenue for stroke management. These findings contribute to the growing understanding of molecular mechanisms in stroke and provide a foundation for developing miR-155-targeted therapies.

Downregulation of PAX1 in OSCC Enhances Stemness and Immunosuppression via IFIT1 and PD‐L1 Pathways

ABSTRACTObjectiveOur study investigated how arecoline‐induced extracellular vesicle (EV) secretion suppresses PAX1 protein production through DNA hypermethylation and examined whether PAX1 downregulation enhances cancer stemness and immunosuppression in the tumor microenvironment.Materials and MethodsEVs were isolated from SAS/TW2.6 cancer cell lines using ultracentrifugation and identified using transmission electron microscopy. PAX1 DNA methylation was tested in an ISO17025‐certified lab, with and without EV pretreatment. Stemness and epithelial–mesenchymal transition markers were assessed by western blotting and 3D culture. PAX1, IFIT1, and PD‐L1 co‐expression were examined through immunofluorescence. Flow cytometry detected various T cells.ResultsArecoline‐induced EVs enhanced PAX1 methylation, suppressing its tumor‐suppressive function. Reduced PAX1 mRNA in OSCCs was linked to larger tumors, nodal metastasis, late‐stage disease, areca quid chewing, and poor survival. Downregulated PAX1 protein negatively correlated with IFIT1 and PD‐L1 expression. Reduced PAX1 promoted stemness via the IFIT1 pathway, increasing PD‐L1 secretion and aiding immune evasion. PD‐L1 expression correlated with Treg and CD8+ T cell levels in OSCC tissues, and the CD4+/CD8+ T cell ratio was lower in OSCC patients than in controls.ConclusionArecoline‐induced EV production, which influences PAX1/IFIT1/PD‐L1 function, may serve as a reliable biomarker for targeted therapy in OSCC patients.

Sodium carboxymethylcellulose disrupts intestinal cell homeostasis by repressing Notch signaling in Drosophila.

Sodium carboxymethylcellulose (CMC), a cellulose derivative, is mainly used in food industry as additives. However, its supplementation has potentially negative effects on organisms. In this study, the effects of CMC on the physiological parameters and intestinal homeostasis in organisms were investigated using Drosophila melanogaster as a model. The results show that CMC did not affect survival and fecundity in flies at three detected concentrations. The puc activity analysis revealed that the CMC did not induce apoptosis in adult midgut. In adults treated with 5% CMC, the midgut contained fewer progenitor cells and showed reduced mitosis compared to control flies. In addition, an increased enteroendocrine (EE) cells ratio was observed in midgut of 5% CMC-fed flies. The RT-qPCR results revealed that notch expression was downregulated in 5% CMC-fed flies, whereas no significant differences in mRNA expression levels were found for genes involved in midgut development. Consistently, the number of Notch positive clones was reduced in flies treated with 5% CMC compared to controls. Taken together, our results suggest that CMC disrupts intestinal cell homeostasis by repressing Notch signaling. These findings provide a new perspective on the effects of CMC on organisms.

Huanglian Ejiao Decoction Alleviates Ulcerative Colitis in Mice Through Regulating the Gut Microbiota and Inhibiting the Ratio of Th1 and Th2 Cells

Huanglian Ejiao Decoction Alleviates Ulcerative Colitis in Mice Through Regulating the Gut Microbiota and Inhibiting the Ratio of Th1 and Th2 Cells

Optimizing gelation time for cell shape control through active learning.

Hydrogels are popular platforms for cell encapsulation in biomedicine and tissue engineering due to their soft, porous structures, high water content, and excellent tunability. Recent studies highlight that the timing of network formation can be just as important as mechanical properties in influencing cell morphologies. Conventionally, time-dependent properties can be achieved through multi-step processes. In contrast, one-pot synthesis can improve both the efficiency and uniformity of cell encapsulation. Reaction kinetics are sensitive to temperatures and pH conditions, thus, monitoring gelation time across different conditions is essential for formulation. In this work, we choose tetra-poly(ethylene glycol) (TPEG) macromers as a model system to examine the relationship between the rate of polymer network formation and cell morphology. Previous studies of this system focused on reactions at neutral pH and room temperature, leaving much of the formulation space underexplored. We use Gaussian process regression (GPR) to minimize response surface errors by strategically selecting additional investigation points based on prior knowledge. Then we extend the knowledge from pre-trained data at neutral pH to a new surface at physiological pH. We find that the gelation time surface can effectively predict the aspect ratio of the encapsulated cells. Additionally, through focal adhesion kinase inhibition, we show that cell shape is influenced by the properties of the forming network in the initial hours as cells develop connections with the matrix. We demonstrate the utility of a high-throughput microrheology approach in enhancing fabrications of synthetic extracellular matrix and cell assemblies.

Phenylbutyric Acid Modulates Apoptosis and ER Stress-Related Gene Expression in Glycogen Storage Disease Type Ib InVitro Model.

Chronic endoplasmic reticulum (ER) stress and increased apoptosis are involved in the pathogenesis of glycogen storage disease Ib (GSD Ib), whereas small molecule phenylbutyrate (4-PBA) showed the capability of reducing ER stress-induced apoptosis. The objective was to generate an invitro system in which capability of small molecules (SMs) to influence ER stress and apoptosis could be screened at the expression level. G6PT-deficient FlpInHEK293 cell line was created and validated using the CRISPR/Cas9 knockout method. Molecular markers of unfolded protein response (ATF4, DDIT3, HSPA5, XBP1s), and apoptosis (BCL2/BAX, CASP3, CASP7) in G6PT-deficient cells were analyzed using RT-qPCR method before and upon the treatment with 4-PBA. Treatment with the most effective dose of 1 mM 4-PBA reduced the expression of UPR markers and executioner caspases, while increased BCL2/BAX ratio in G6PT-deficient cells. Our results proved the concept that 4-PBA could alleviate markers of ER stress detected in the GSD Ib invitro model system and prevent cell death. This cost-effective invitro model screens the therapeutic potential of SMs affecting ER stress and apoptosis in G6PT-deficient kidney cells, offering a first-line screening assay for promising compounds. 4-PBA's potential repurposing for GSD Ib patients opens new research directions.

Differentiating multiple sclerosis with predominant spinal cord and optic nerve involvement from other autoimmune demyelinating diseases using B cell immunophenotyping and gene expression profiling.

Multiple sclerosis (MS) may present with predominant involvement of the spinal cord and optic nerve (MS/w-SCON) and mimic other autoimmune inflammatory demyelinating disorders (AIDD) such as neuromyelitis optica spectrum disorder (NMOSD), and relapsing inflammatory optic neuritis (RION). Thus, biomarkers are required for effective differential diagnosis of AIDD. Patients with MS/w-SCON (n = 20), MS without involvement of SCON (MS/wo-SCON) (n = 22), NMOSD (n = 16), RION (n = 15) and healthy individuals (n = 21) were included. Peripheral blood cell immunophenotypes were analyzed by flow cytometry and gene expression levels of isolated B cells were assessed by microarray analysis and quantitative real-time PCR. Significantly increased Breg, plasmablast, and plasma cell ratios distinguished MS/w-SCON from other groups. Most differentially expressed B cell genes were subjected to protein-protein interaction yielding a network of 13 immune mediators (IL18, IL18R1, P2RY12, CSF3R, TNFSF4, TNFRSF13C, TNFRSF1A, CCL20, CCL5, CCR4, CCL4L2, CXCL5, CXCL10). CCL4L2 and CCR4 expression levels distinguished MS/w-SCON. IL18/IL18R1 and CCL5 were distinguishing factors for NMOSD and RION, respectively. Peripheral blood B cell expression levels of CCL4L2, CCR4, IL18, IL18R1 and CCL5 are candidate biomarkers for differential diagnosis of AIDD of CNS. Our results substantiate the significance of B cells in the pathogenesis of these disorders.

FT538, iPSC-derived NK cells, enhance AML cell killing when combined with chemotherapy.

Induced pluripotent stem cell (iPSC)-derived natural killer (NK) cells offer an opportunity for a standardized, off-the-shelf treatment with the potential to treat a wider population of acute myeloid leukaemia (AML) patients than the current standard of care. FT538 iPSC-NKs express a high-affinity, noncleavable CD16 to maximize antibody dependent cellular cytotoxicity, a CD38 knockout to improve metabolic fitness, and an IL-15/IL-15 receptor fusion preventing the need for cytokine administration, the main source of adverse effects in NK cell-based therapies. Here, we sought to evaluate the potential of FT538 iPSC-NKs as a therapy for AML through their effect on AML cell lines and primary AML cells. We observed that FT538 iPSC-NKs induce effector-to-target cell ratio dependent apoptosis in cell lines and primary AML cells, including cells from high-risk patients. Flow cytometric analysis revealed that FT538 iPSC-NKs induce AML cell death when combined with the AML therapies: cytarabine, venetoclax and gilteritinib. Moreover, cytarabine did not affect FT538 iPSC-NK viability, suggesting that iPSC-derived NK therapies and chemotherapy may be a promising treatment combination. This study provides the basis for further study of iPSC-derived NK cell therapies as a treatment option for high-risk AML patients, particularly those with disease resistant to standard therapies.

Prediction of deep vein thrombosis in pregnant women by platelet indices: A retrospective case-control study.

This study aimed to investigate the predictive value of the platelet indices for the diagnosis of acute deep vein thrombosis (DVT) in pregnant women. Pregnant women aged 20-45 years who presented to the emergency room with leg pain and edema and underwent bilateral lower extremity venous Doppler examination for DVT were included in the study. Platelet large cell ratio (P-LCR), platelet distribution width (PDW) and mean platelet volume (MPV) values, which provide information on current platelet activity in the hemogram parameters recorded at first presentation in the emergency department, were analyzed between the two groups. Pregnant women with a history of venous thromboembolism, anticoagulant medication, heart disease, cancer, essential thrombocytopenia, gestational thrombocytopenia, pulmonary embolism, and multiple pregnancies were excluded. A total of 125 participants were included in the study. Acute DVT was diagnosed in 37 participants using venous Doppler of the lower extremities. In the remaining 88 patients, the diagnosis of DVT was excluded. There was no significant difference between the groups in terms of age, body mass index, parity, and gestational age at diagnosis. Although platelet counts were similar between groups, P-LCR, PDW, and MPV values, which provide information on platelet shape and volume, were statistically significantly lower in Group DVT (p < 0.05). According to this study, low MPV, PDW and P-LCR values in pregnant women can be evaluated as independent markers in the diagnosis of acute DVT or these markers can be included in the DVT scoring system for pregnant women.