To establish and optimize a protocol for the efficient isolation, identification, and short-term culture of primary CD34⁺ acute myeloid leukemia (AML) cells from bone marrow (BM) and peripheral blood (PB) samples of newly diagnosed AML patients, providing a stable primary cell model for basic and translational research on AML.

BM and PB samples were collected from 10 newly diagnosed AML patients. Cells were enriched using Ficoll density gradient centrifugation combined with CD34⁺ immunomagnetic bead sorting. Sorting efficiency was evaluated by flow cytometry via CD34⁺/CD117⁺ immunophenotyping, and cell morphology was observed using Wright-Giemsa staining. Cell counts and CCK-8 assays were employed to dynamically monitor the proliferation and viability of BM- and PB-derived cells over 72 hours. Furthermore, the effects of 10% versus 20% fetal bovine serum (FBS) concentrations on the proliferation of BM-derived cells were compared.

Following immunomagnetic sorting, the purity of CD34⁺ cells from both BM and PB exceeded 90% (92.7%±2.7% for BM, 91.1%±3.0% for PB, P>0.05), with a high proportion of CD117⁺ co-expression (71.9%±3.3% for BM, 72.6%±3.0% for PB, P>0.05). The cells exhibited typical primitive myeloid morphology. During the 72-hour culture period, the peak proliferation at 48 hour (approximately 4.0×10³±0.2×10³ viable cells) and metabolic activity (OD₄₅₀ value of 1.59±0.07) of BM-derived cells were significantly higher than those of PB-derived cells (3.2×10³±0.1×10³, 1.14±0.08, both P<0.01), with superior morphological stability in later stages. No statistically significant difference was observed between 10% and 20% FBS regarding short-term cell proliferation, although 20% FBS showed a slight advantage in maintaining cell morphology.

This study successfully established a stable and efficient system for the isolation and short-term culture of human primary CD34⁺ AML cells. This system is applicable to both BM and PB samples. The results demonstrate that BM-derived cells possess stronger in vitro proliferative capacity and that 10% FBS is sufficient for their short-term culture requirements.

To investigate the expression level of UBE2QL1 in multiple myeloma (MM) and its correlation with disease progression and prognosis, as well as to elucidate the biological role of UBE2QL1 in MM through in vitro cell experiments.

Bioinformatics analysis was used to explore the expression level of UBE2QL1 in MM and its impact on prognosis. Concurrently, a polyclonal antibody against UBE2QL1 was synthesized, and its specificity was confirmed through mass spectrometry. The expression level of UBE2QL1 in MM cell lines and primary bone marrow samples was validated using qRT-PCR and Western blot, followed by comparative analysis with the corresponding clinical data. Subsequently, the UBE2QL1 gene was knocked down in the MM cell lines NCI-H929 and MM.1S. to verify its regulatory effects on MM cell functions.

Analyses based on the Hemap, DepMap (CCLE), and GEO databases revealed that UBE2QL1 was highly expressed in MM, and its expression level was closely associated with disease progression and poor prognosis.In vitro experiments further confirmed the elevated expression of UBE2QL1 in MM cell lines and primary bone marrow samples from patients, showing a positive correlation with the proportion of plasma cells. Knockdown of UBE2QL1 in MM cell lines inhibited cell proliferation, promoted cell apoptosis, and improved cellular sensitivity to widely used therapeutic agents, including bortezomib, carfilzomib, and lenalidomide.

UBE2QL1 is highly expressed in MM and is closely associated with adverse disease progression and poor prognosis. Knockdown of UBE2QL1 in vitro can effectively inhibit MM cell proliferation, promote apoptosis, and enhance cellular sensitivity to therapeutic agents. Monitoring the expression level of UBE2QL1 may help to more accurately assess the prognosis of MM patients and provide a reference for therapeutic decision-making.

To explore the application of multiparameter flow cytometry combined with serum cystatin C (Cys C) and interleukin-10 (IL-10) in the diagnosis of multiple myeloma (MM).

A total of 167 patients with suspected MM attending our hospital from March 2023 to July 2024 were enrolled, including 106 MM patients (MM group) and 61 non-MM patients (non-MM group). The patients underwent multiparameter flow cytometry and serum Cys C and IL-10 detection. The multivariate logistic regression analysis was conducted to identify the influencing factors of MM, and ROC curve analysis to evaluate the diagnostic value of multiparameter flow cytometry combined with serum Cys C and IL-10 for MM.

The ratio of plasma cells, CD56, the levels of serum Cys C and IL-10 in the MM group were significantly higher than those in the non-MM group, while the levels of CD19 and CD45 were lower (all P < 0.001). Multivariate logistic regression analysis showed that the ratio of plasma cells, Cys C, and IL-10 were risk factors for MM (OR =2.493, 3.843, 2.726, P < 0.05). ROC curve analysis indicated that the diagnostic value of multiparameter flow cytometry combined with serum Cys C and IL-10 for MM was higher than that of single diagnosis.

The ratio of plasma cells and the levels of serum Cys C and IL-10 are elevated in MM patients, which lead to an increased risk of MM disease. The combination of the three has certain diagnostic value for MM.

To investigate the effects of alantolactone (AL) on the proliferation and apoptosis of acute myeloid leukemia (AML) cells and its underlying molecular mechanisms.

AML cell lines SHI-1 and THP-1 were treated with various concentrations of AL. Cell proliferation capacity was assessed using CCK-8 assay, and cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Intracellular reactive oxygen species (ROS) levels were measured using DCFH-DA fluorescent probe, while the mitochondrial membrane potential (MMP) was evaluated using tetramethylrhodamine ethyl ester (TMRE). Western blot analysis was performed to detect the expression of cell cycle regulatory proteins (CDK4, CDK6), apoptosis-related proteins (cleaved caspase-3, Bax, Bcl-XL, PARP), and proteins involved in the PI3K/Akt signaling pathway.

AL significantly inhibited the proliferation of SHI-1 and THP-1 cells in a concentration-dependent (r _SHI-1=-0.942 2, r _THP-1=-0.882 6) and time-dependent (r _SHI-1=-0.979 3, r _THP-1=-0.940 0) manner. Meanwhile, AL treatment significantly increased the proportion of cells in G₀/G₁ phase and apoptosis rate of both SHI-1 and THP-1 cells (P < 0.05), and downregulated the expression of CDK4 and CDK6 (P < 0.05). Additionally, AL significantly upregulated Bax protein expression, downregulated Bcl-XL protein expression, activated caspase-3 and cleaved PARP, thereby promoting cell apoptosis (all P < 0.05). Furthermore, AL treatment significantly elevated intracellular ROS levels and decreased MMP (P < 0.05). Further studies demonstrated that AL could induce programmed cell death in AML cells by inhibiting the activation of the PI3K/Akt signaling pathway.

AL may exert anti-leukemic effects by suppressing AML cell proliferation and inducing apoptosis through multiple mechanisms, including inhibition of PI3K/Akt signaling pathway activation, modulation of Bcl-2 family protein expression, and induction of intracellular oxidative stress.

We investigated the role of tumor microenvironment (TME) in evolution of liver cirrhosis to hepatocellular carcinoma (HCC). Nonmalignant hepatic tissues, including chronic hepatitis and cirrhosis, retained expression of liver specific miR-122 and hepatocyte nuclear factor 4 (HNF-4) transcription factor and exhibited an immune surveillance program characterized by Inducible T cell Costimulatory Ligand (ICOSL) and NF-κB expression, low Suppressor of Cytokine Signaling 1 (SOCS1) levels, and abundant cytotoxic T cell infiltration. In contrast, progression to HCC was associated with loss of cytotoxic T cells, loss of ICOSL, PDL1, HNF-4, SMAD7, and NF-κB expression, and marked upregulation of YAP1 oncogene. TGF-β overexpression was induced by SMAD7 loss, mediated by miR-21 upregulation. Loss of HNF4, was associated with reduced miR-122 expression and subsequent upregulation of its target, SOCS1. Chronic inflammation, driven by miR-155, created an immune permissive environment that facilitated miR-21 induction and miR-122 loss. In addition, miR-21 and miR-155 contributed to malignant transformation through suppression of numerous tumor suppressor genes and enhanced YAP1 expression, a potent hepatocyte oncogene. These findings were recapitulated in orthotopic HCC mouse models, which demonstrated loss of ICOSL, PDL1, and HNF4 expression, minimal cytotoxic T cell infiltration, and increased of SOCS1, YAP1, and TGF-β expression. It is concluded that the transition from cirrhosis to HCC is associated by coordinated immunosuppression and oncogenic signaling driven by miR-21 and miR-155, together with the loss of miR-122. Targeting miR-155 may therefore represent a preventive strategy for HCC development in patients with chronic viral hepatitis.

Breast cancer (BC) remains the most common malignancy among women worldwide. miR-199a-3p has been implicated in several cancers, while Cyclin-Dependent Kinase 7 (CDK7) plays a critical role in cell cycle regulation and transcriptional control.

To evaluate the expression profile and diagnostic performance of miR-199a-3p and CDK7 in BC tissues and investigate their associations with clinicopathological characteristics.

This study was conducted on 30 paired BC and adjacent normal breast tissue samples collected in Erbil, Iraq. Expression levels of miR-199a-3p and CDK7 were quantified using qRT-PCR. Also, blood samples were collected to estimate CA 15 - 3.

Among 30 BC patients, there was a significant difference in miR-199a-3p expression in tumor tissues compared to paired control tissues (P = 0.0029), with a downregulation of 0.506-fold relative to the control group. In contrast, CDK7 expression did not differ significantly between tumor and control tissues (P = 0.140). The serum CA 15 - 3 levels were considerably higher in BC patients (P < 0.001) and the AUC was 0.754 (95% CI: 0.623-0.867) with a sensitivity of 43.3% and specificity of 96.7% at a threshold value of 25.1 U/mL. miR-199a-3p presented an AUC of 0.720 (95% CI, 0.581-0.839) and CDK7 an AUC of 0.621 (95% CI, 0.467-0.764). The combined miR-199a-3p/CDK7 model showed an AUC of 0.750 (95% CI: 0.619-0.886) with 76.7% sensitivity and 80.0% specificity. The higher CDK7 expression was associated with PR-negative status (P = 0.038).

miR-199a-3p may be a possible biomarker for BC diagnosis with considerably low expression and moderate diagnostic efficacy. The combination miR-199a-3p/CDK7 model enhanced diagnostic discrimination, while serum CA 15 - 3 showed good specificity. The relationship of CDK7 with PR status further supports its therapeutic importance.

Uterine leiomyoma (UL) is a prevalent tumour of the female reproductive system which is poorly understood in terms of pathogenesis, and treatment options are limited. MicroRNAs (miRNAs) have been identified as being associated with tumorigenesis. The aim of the study is to elucide the function of miR-483-5p in promoting UL progression by targeting TIMP metallopeptidase inhibitor 2 (TIMP2) and its underlying molecular mechanisms.

The study included 340 patients with UL. The expression level of miR-483-5p was measured using quantitative real-time PCR (qPCR). Following knockdown of miR-483-5p in uterine leiomyoma cells (UtLMCs), cell proliferation, migration, and apoptosis were assessed using CCK-8, Transwell, and flow cytometry assays. Subsequently, bioinformatics analysis and luciferase reporter gene detection identified the target of miR-483-5p.

Compared to the adjacent myometrium, miR-483-5p was upregulated in UL tissues. Experiments revealed that miR-483-5p expression levels were increased in UtLMCs. It was determined that TIMP2 is the direct target of miR-483-5p. Furthermore, miR-483-5p significantly promoted the proliferation and migration of UtLMCs, and inhibited their apoptosis by targeting TIMP2.

Collectively, these findings reveal the molecular mechanism by which miR-483-5p negatively regulates TIMP2 to promote UL development, providing new insights for the targeted therapy of UL.

MicroRNAs (miRNAs) are essential in the modulation of cellular activities. Studies suggest that miR-3615 may possess a tumour-suppressive function in lung adenocarcinoma (LUAD); however, its precise role and molecular mechanisms in LUAD progression remain incompletely understood. The aim of the study is to systematically investigate the biological functions and underlying mechanisms of miR-3615 in LUAD.

The expression of miR-3615 in LUAD cell lines was quantified by real-time quantitative PCR (RT-qPCR). Subsequently, its functional effects on proliferation, migration, invasion, and apoptosis were evaluated in A549 and H1299 cells using the CCK-8 assay, Transwell assay, and flow cytometry, respectively. Potential target genes of miR-3615 were predicted and functionally annotated using bioinformatics methods. The binding interaction of miR-3615 with the CALML4 gene was demonstrated via a dual-luciferase reporter assay.

miR-3615 expression was notably downregulated in LUAD cell lines compared with normal cells. Overexpression of miR-3615 inhibited the proliferation, migration, and invasion capabilities of LUAD cells while promoting apoptosis. Integrated bioinformatics analysis involving target prediction and functional enrichment identified CALML4 as a key candidate target gene of miR-3615. CALML4 was confirmed as a direct target of miR-3615. The effects of miR-3615 mimic on LUAD cell proliferation, migration, invasion, and apoptosis were partly attenuated by CALML4 overexpression.

miR-3615 exerts its functional impact on LUAD cells by directly downregulating CALML4, thereby curbing proliferation, migration, and invasion and promoting apoptosis. The elucidated miR-3615/CALML4 axis advances the theoretical foundation for studying LUAD development.

Circular RNAs (circRNAs) are recognized as critical regulators of gastric cancer (GC) progression. Nevertheless, the biological function of circ_0088302 in gastric cancer and its epitranscriptomic regulatory mechanism remain unclear.

The expression of circ_0088302 was analyzed based on plasma circRNA data from GSE93541 and RT-qPCR, and its subcellular localization was determined by RNA-FISH. Cellular malignant phenotypes were evaluated using CCK-8 assay, colony-formation assay, wound-healing assay, Transwell assay, and flow cytometry. MeRIP-qPCR, RIP, dual-luciferase reporter assay, and actinomycin D assay were performed to assess m⁶A modification, IGF2BP1 binding, candidate m⁶A sites, and RNA stability. AVJ16 was used to inhibit IGF2BP1 function.

circ_0088302 was highly expressed in plasma samples from GC patients and GC cells. Knockdown of circ_0088302 repressed cell viability, colony formation, and migration, and promoted cell apoptosis. circ_0088302 exhibited m⁶A enrichment and bound to IGF2BP1, and the predicted 292 site was involved in IGF2BP1-dependent regulation. IGF2BP1 knockdown reduced circ_0088302 levels and accelerated its degradation. AVJ16 suppressed the viability of GC cells and abrogated the malignant phenotypes induced by circ_0088302 overexpression.

IGF2BP1 may facilitate the malignant phenotypes of GC cells by maintaining the RNA stability of m⁶A-modified circ_0088302, suggesting that the IGF2BP1/circ_0088302 axis may represent a potential therapeutic target for GC.

Lung transplantation is a complex therapy for end stage lung disease with variable postoperative outcomes. Textbook outcomes, defined by freedom from perioperative morbidity and mortality, have emerged as a comprehensive measure of surgical success. Body mass index (BMI) is commonly used to assess nutritional status but does not distinguish between skeletal muscle and adipose tissue. CT-based body composition analysis can differentiate these tissues and may provide a more precise assessment of physiologic reserve in lung transplantation.

We performed a retrospective cohort study of 377 adults undergoing single or bilateral lung transplantation at a single academic institution (2019-2023). Preoperative abdominal CT scans were used to quantify skeletal muscle, subcutaneous fat, and visceral fat areas. A composite textbook outcome was defined as freedom from intraoperative complications; postoperative reintervention; readmission, acute rejection, and dialysis within 30 days; 90-day mortality; primary graft dysfunction; ECMO at 72 h; tracheostomy within 7 days; reintubation; and extubation >48 h. Multivariable logistic regression assessed associations between body composition metrics and lung transplant outcomes after adjusting for potential confounders.

Among 377 recipients, 108 (28.6%) achieved textbook outcome. Increased skeletal muscle area was associated with increased odds of achieving textbook outcome (OR per 50cm²: 1.808 [1.061-3.119]) and decreased odds of 1-year mortality (OR per 50cm²: 0.423 [0.195-0.890]). Subcutaneous fat area, visceral fat area, and BMI were not significantly associated with outcomes.

CT-based skeletal muscle was associated with textbook outcome and 1-year mortality, whereas BMI was not. CT-based body composition analysis may be a promising tool for lung transplant risk stratification.