Biological Procedures Online最新文献

Background: Myelodysplastic neoplasms (MDS) are a category of myeloid malignancies that can progress to acute myeloid leukemia (AML). Arsenic sulfide (As₂S₂) has demonstrated efficacy in the treatment of MDS. However, further investigation is required to elucidate its specific therapeutic targets.

Objective: To examine the influence of ABI2 on MDS and clarify the regulatory role of As₂S₂ on ABI2 within the therapeutic framework of MDS.

Methods: To explore ABI2's role in the expression and prognosis of patients with hematological malignancies, RNA interference was used to reduce ABI2 expression in SKM-1, K562 and MDS-L cell lines. The therapeutic efficacy of As₂S₂ and its regulatory impact on ABI2 were evaluated both in vitro and in vivo.

Results: ABI2 was found to be overexpressed in the peripheral blood of patients with MDS and AML, which was significantly associated with decreased survival rates in AML patients. Inhibition of ABI2 decreased the proliferation and increased the apoptosis of SKM-1, K562 and MDS-L cells. As₂S₂ also inhibited cell proliferation and induced apoptosis in these cells, but its efficacy diminished significantly with ABI2 silencing. Additionally, As₂S₂ showed therapeutic benefits in NHD13 mice by downregulating bone marrow ABI2 expression.

Conclusion: The collective findings suggest that ABI2 is significantly overexpressed in MDS and AML, and its expression was correlated with unfavorable prognostic outcomes. Furthermore, As₂S₂ demonstrates therapeutic efficacy in MDS by modulating the expression levels of ABI2.

Background: Gastric cancer remains a major global health concern with approximately one million new cases annually. Despite five decades of extensive research, comprehensive analysis mapping gastric carcinogenesis research evolution and emerging therapeutic frontiers remains limited. This study aims to systematically analyze the intellectual landscape, research trends, and future directions in gastric carcinogenesis research.

Materials and methods: We analyzed 11,527 scientific publications on gastric carcinogenesis from 1970-2024 using advanced scientometric tools including CiteSpace and VOSviewer for co-citation analysis, collaboration network mapping, and keyword co-occurrence analysis.

Results: Publication output demonstrated exponential growth, peaking at 547 publications in 2022. Asia leads research productivity with China, Japan, and South Korea contributing 37.50% of total publications, while the United States exhibits highest collaborative influence with betweenness centrality of 0.45. Seven distinct research clusters emerged encompassing Helicobacter pylori pathogenesis, molecular mechanisms, precancerous lesions, autoimmune gastritis, and therapeutic interventions. Five emerging trends were identified: stromal microenvironment interactions in carcinogenesis progression, gastric organoid models for disease modeling, artificial intelligence-enhanced endoscopic detection technologies, biomarker development for early diagnosis, and traditional herbal medicine applications for chemoprevention.

Conclusions: Research gaps persist in autoimmune gastritis pathways and clinical translation of molecular discoveries. Emerging research directions encompass advanced modeling systems, molecular diagnostics, and technological applications, though their clinical impact remains to be established through future validation studies.

Objective: This study aims to evaluate the clinical efficacy of combining spleen‑nourishing and qi‑tonifying therapy with programmed cell death protein 1 (PD‑1)/programmed death‑ligand 1 (PD‑L1) inhibitors in patients with stage IIIB-IV non‑small cell lung cancer (NSCLC), and to develop a survival prediction model based on progression‑free survival (PFS) and overall survival (OS).

Methods: A retrospective cohort of 246 patients with stage IIIB-IV NSCLC receiving immunotherapy at the Tumor Medicine Center of the First Hospital of Hunan University of Chinese Medicine from September 2019 to October 2024 is assembled. Patients are categorized into a traditional Chinese medicine (TCM) group (n = 174) or a control group (n = 72) according to use of TCM decoctions. Baseline clinical and pathological characteristics and treatment data are collected. Cox proportional hazards regression is used to analyze survival‑related factors. Kaplan-Meier curves and log‑rank tests are used to compare survival. Logistic regression is used to identify factors associated with the objective response rate (ORR). Survival prediction models based on PFS and OS are constructed and validated.

Results: Median PFS is 5.9 months in the TCM group versus 2.9 months in the control group (P < 0.001), and median OS is 22.3 versus 14.8 months (P < 0.001). On multivariable analysis, TCM intervention, PD‑L1 expression, alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and cytokeratin fragment 21‑1 (CYFRA21‑1) are independent predictors of PFS; TCM intervention, brain metastasis, and carcinoembryonic antigen (CEA) independently influence OS. The 6‑month PFS rate, 1‑year PFS rate, and 1‑year OS rate are significantly higher in the TCM combination group than in controls (P < 0.05). TCM intervention and sex are independent predictors of ORR (P < 0.05). Model validation shows areas under the curve (AUCs) of 0.705 (PFS) and 0.636 (OS) in the prediction-model group, and 0.879 (PFS) and 0.765 (OS) for internal validation.

Conclusion: The addition of TCM to PD‑1/PD‑L1 inhibitors prolongs PFS and OS in patients with stage IIIB-IV NSCLC. A survival prediction model based on routine clinical characteristics demonstrates predictive utility, offering a potential tool to inform clinical decision‑making.

Background: Membrane-bound vesicles called apoptotic bodies (ApoBDs) formed during apoptosis are being recognized as non-invasive biomarkers for neurological diseases like dementia and ischemic stroke. Centrifugation-based isolation of these vesicles from blood was suggested in a recent paper by Serrano-Heras et al. (2020), which asserted their use for clinical diagnosis. This initial study provides a foundational methodology for ApoBD diagnostics. Albeit, centrifugation parameters employed present technical challenges that require optimization to ensure the purity and reproducibility of ApoBD preparations for clinical utility.

Results: Due to the use of improper centrifugation speeds (160×g and 700×g) not consistent with accepted norms for the separation of platelets (ideally 2,000-5,000×g), running the risk of being seriously contaminated by platelets and platelet-derived EVs (P-EVs), which make up 70-90% of EVs in circulation. Such contamination can render functional studies problematic, cloud proteomic and size-distribution analysis, and lower the specificity of ApoBDs enumeration on the basis of Annexin V/PI labeling. Even more extreme centrifugation has little chance to eliminate platelets completely, according to comparative literature data. Increased centrifugal pressure, density gradient separation, size-exclusion chromatography (SEC), and confirmation with platelet-specific markers are among some procedural changes recommended to improve sample purity in attempts to solve such problems.

Conclusions: Even as the Serrano-Heras et al. study pinpoints the potential of ApoBDs as biomarkers, constraints to the procedure under consideration pose the threat of bias in future studies. Proper isolation of ApoBDs requires multimodal confirmation, optimized platelet removal protocols and clinically feasible methods like SEC are essential to enhance ApoBDs isolation for neurological disease diagnostics. Rectification of such issues enhance significantly the practicality of ApoBDs in clinical practice for the detection of neurological illnesses.

Background: Western blotting is a fundamental technique for protein detection and quantification, with critical dependence on antibody for its sensitivity and specificity. Antibody solutions are frequently reused to conserve the limited stock and compensate for their high cost. However, storing antibody solutions is often challenged by protein aggregation and loss of immunoreactivity, especially when skim milk is added. Recent advancements in Western blot include microfluidic techniques which minimize antibody consumption. To facilitate these techniques, a storage strategy is needed for diluted antibody solutions to properly preserve their immunoreactivity.

Results: We found that Tris-buffered saline with 0.1% Tween 20 (TBST), an ordinary washing buffer for Western blot, provides protection to diluted antibodies for extended period of storage at 4 °C. We discovered that TBST-diluted commercial antibodies retained their immunoreactivity for 281 days at ambient temperature and 1,690 days at 4 °C. In addition, we demonstrated that Tween 20 plays a critical role in stabilizing the diluted antibody.

Conclusions: We propose that TBST is an effective and practical buffer to preserve diluted antibodies, which can be utilized as an integral part of strategies to reduce antibody consumption in laboratories.

Background: Advances in nanomedicine have spurred interest in antibody-conjugated carbon nanotubes for targeted cancer therapy. CD133, a marker enriched in chemoresistant cancer stem cells, presents a strategic target for precision drug delivery. This study explores functionalized single-walled carbon nanotubes (SWCNTs) as dual-action platforms for enhanced doxorubicin (DXR) delivery and CD133-specific targeting.

Methods: Two nano formulations-SWCNT-Ab/DXR (antibody-functionalized) and SWCNT-PEG-Ab/DXR (pegylated antibody-functionalized)-were engineered to encapsulate DXR. Release profiles, cytotoxicity, and apoptosis were assessed in CD133⁺ HT-29 colorectal cancer cells and CD133^- CHO control cells. Computational modeling included DFT-D structural optimization, Monte Carlo adsorption simulations for DXR binding analysis, and molecular docking to evaluate carrier-receptor interactions.

Results: PEGylation markedly enhanced colloidal stability and drug-loading capacity, with SWCNT-PEG-Ab/DXR achieving 92% DXR encapsulation vs. 78% for non-PEGylated counterparts. In vitro, the PEGylated system showed amplified cytotoxicity (IC50: 2.1 µM vs. 3.8 µM for SWCNT-Ab/DXR) and 1.7-fold higher apoptosis induction in HT-29 cells. Computational data aligned with experimental findings: DXR adsorption energy was strongest on PEG-NH₂-modified SWCNTs (- 32.6 kcal/mol) versus carboxylated variants (- 24.8 kcal/mol), confirming PEG's role in stabilizing drug-carrier interactions.

Conclusions: By integrating PEG-mediated stealth properties, charge-modified surfaces, and antibody targeting, SWCNT-PEG-Ab/DXR emerges as a multifunctional nanoplatform with enhanced tumor selectivity and therapeutic payload delivery. This dual experimental-computational approach underscores the potential of rationally engineered nanotubes to overcome limitations in conventional chemotherapy.