Mitochondria-Specific Molecular Crosstalk between Ferroptosis and Apoptosis Revealed by In Situ Raman Spectroscopy.

Ferroptosis and apoptosis are two types of regulated cell death that are closely associated with the pathophysiological processes of many diseases. The significance of ferroptosis-apoptosis crosstalk in cell fate determination has been reported, but the underlying molecular mechanisms are poorly understood. Herein mitochondria-mediated molecular crosstalk is explored. Based on a comprehensive spectroscopic investigation and mass spectrometry, cytochrome c-involved Fenton-like reactions and lipid peroxidation are revealed. More importantly, cytochrome c is found to induce ROS-independent and cardiolipin-specific lipid peroxidation depending on its redox state. In situ Raman spectroscopy unveiled that erastin can interrupt membrane permeability, specifically through cardiolipin, facilitating cytochrome c release from the mitochondria. Details of the erastin-cardiolipin interaction are determined using molecular dynamics simulations. This study provides novel insights into how molecular crosstalk occurs around mitochondrial membranes to trigger ferroptosis and apoptosis, with significant implications for the rational design of mitochondria-targeted cell death reducers in cancer therapy.

In Situ Raman Spectroscopy Reveals Cytochrome c Redox-Controlled Modulation of Mitochondrial Membrane Permeabilization That Triggers Apoptosis.

The selective interaction of cytochrome c (Cyt c) with cardiolipin (CL) is involved in mitochondrial membrane permeabilization, an essential step for the release of apoptosis activators. The structural basis and modulatory mechanism are, however, poorly understood. Here, we report that Cyt c can induce CL peroxidation independent of reactive oxygen species, which is controlled by its redox states. The structural basis of the Cyt c-CL binding was unveiled by comprehensive spectroscopic investigation and mass spectrometry. The Cyt c-induced permeabilization and its effect on membrane collapse, pore formation, and budding are observed by confocal microscopy. Moreover, cytochrome c oxidase dysfunction is found to be associated with the initiation of Cyt c redox-controlled membrane permeabilization. These results verify the significance of a redox-dependent modulation mechanism at the early stage of apoptosis, which can be exploited for the design of cytochrome c oxidase-targeted apoptotic inducers in cancer therapy.

In Situ and Real-Time Monitoring of Mitochondria-Endoplasmic Reticulum Crosstalk in Apoptosis via Surface-Enhanced Resonance Raman Spectroscopy.

The crosstalk between mitochondria and endoplasmic reticula plays a crucial role in apoptotic pathways in which reactive oxygen species (ROS) produced by microsomal monooxygenase (MMO) are believed to accelerate cytochrome c release. Herein, we successfully demonstrate the potential of surface-enhanced resonance Raman spectroscopy (SERRS) for monitoring MMO-derived ROS formation and ROS-mediated cytochrome c release. Silver nanoparticles coated with nickel shells are used as both Raman signal enhancers and electron donors for cytochrome c. SERRS of cytochrome c is found to be sensitive to ROS, allowing for in situ probing of ROS formation with a cell death inducer. Label-free evaluation of ROS-induced apoptosis is achieved by SERRS-based monitoring of cytochrome c release in living cells. This study verifies the capability of SERRS for label-free, in situ, and real-time monitoring of the mitochondria-endoplasmic reticulum crosstalk in apoptosis and provides a novel strategy for the rational design and screening of ROS-inducing drugs for cancer treatment.

Poor prognosis, hypomethylation, and immune infiltrates are associated with downregulation of INMT in head and neck squamous cell carcinoma.

Background: Indiolethylamine-N-methyltransferase (INMT) is a methyltransferase responsible for transferring methyl groups from methyl donor SAM to its substrate. S-adenosyl-l-methionine (SAM), obtained from the methionine cycle, is a naturally occurring sulfonium compound that is vital to cellular metabolism. The expression of INMT is down-regulated in many tumorous tissues, and it may contribute to tumor invasion and metastasis. Nevertheless, the expression of INMT and its relationship to methylation and immune infiltrates in head and neck squamous cell carcinoma (HNSC) remains a mystery. Thus, we evaluated expression, clinicopathological features, prognosis, several critical pathways, DNA methylation, and immune cell infiltration for the first time. Methods: Analysis of the clinicopathological characteristics of INMT expression, several tumor-related bioinformatics databases were utilized. In addition, the role of INMT expression was analyzed for prognosis. Several INMT-related pathways were enriched on the LinkedOmics website. In addition, we have analyzed the methylation of INMT in HNSC in detail by using several methylation databases. Lastly, the relationship between INMT gene expression and immune infiltration was analyzed with ssGSEA, Timer, and TISIDB. Results: In HNSC, mRNA and protein levels were significantly lower than in normal tissues. The low expression of INMT was statistically associated with T stage, histological grade, gender, smoking history, and alcohol consumption. HNSC patients with low INMT expression have a poorer OS (overall survival) compared to those with high levels of expression. In addition, the multivariate analysis revealed INMT expression to be a remarkable independent predictor of prognosis in HNSC patients. An analysis of gene enrichment showed that several pathways were enriched in INMT, including the Ras signaling pathway, the cGMP-PKG signaling pathway, and others. Moreover, methylation patterns of INMT detected in a variety of methylation databases are closely associated with mRNA expression and prognosis. Finally, INMT was significantly correlated with immune infiltration levels. Conclusion: HNSC with low levels of INMT exhibits poor survival, hypomethylation, and immune infiltration. For HNSC, this study presented evidence that INMT is both a biomarker of poor prognosis and a target of immunotherapy.

Fabrication and characterization of a novel Ba2+-loaded sawdust biochar doped with iron oxide for the super-adsorption of SO42- from wastewater.

Biochar is a low-cost adsorbent used in the treatment of contaminated wastewater. We investigated the potential of an Fe-impregnated, Ba2+-loaded biochar (Fe-(Ba-BC)) for the removal of SO42- from aqueous solutions. The Ba2+-loaded biochar was synthesized from sawdust impregnated with iron oxide via pyrolysis at 600 °C. The porous structure of the Fe-(Ba-BC) was identified by scanning electron microscopy before sulfate was adsorbed onto the adsorbent. Functional groups were determined by energy-dispersive spectrophotometry and Raman spectrometry.. The Fe-(Ba-BC) Raman peaks before the experiment were higher than after, suggesting the precipitation of BaSO4. The presence of BaCl2 on the surface of the biochar was confirmed by X-ray diffraction. Batch sorption results showed that Fe-(Ba-BC) strongly adsorbed aqueous SO42- with a removal efficacy of 96.7% under the optimum conditions of 0.25 M BaCl2, a contact time of 480 min, a pH of 9 and an adsorbent dose of 2 g. The optimum condition for removal and reaction rate kinetics analysis indicated that adsorption curve fitted well with PSO, k2 0.00015 confirmed the removal of SO42- via chemisorption. Thus, Fe-(Ba-BC) was found to be a favorable adsorbent for removing SO42-.

Electron transfer between cytochrome c and microsomal monooxygenase generates reactive oxygen species that accelerates apoptosis.

Generation of reactive oxygen species (ROS) are possibly induced by the crosstalk between mitochondria and endoplasmic reticula, which is physiologically important in apoptosis. Cytochrome c (Cyt c) is believed to play a crucial role in such signaling pathway by interrupting the coupling within microsomal monooxygenase (MMO). In this study, the correlation of ROS production with the electron transfer between Cyt c and the MMO system is investigated by resonance Raman (RR) spectroscopy. Binding of Cyt c to MMO is found to induce the production of ROS, which is quantitatively determined by the in-situ RR spectroscopy reflecting the interactions of Cyt c with generated ROS. The amount of ROS that is produced from isolated endoplasmic reticulum depends on the redox state of the Cyt c, indicating the important role of oxidized Cyt c in accelerating apoptosis. The role of electron transfer from MMO to Cyt c in the apoptotic mitochondria-endoplasmic reticulum pathway is accordingly proposed. This study is of significance for a deeper understanding of how Cyt c regulates apoptotic pathways through the endoplasmic reticulum, and thus may provide a rational basis for the design of antitumor drugs for cancer therapy.

[Application of objective performance criteria in post-marketing evaluation schemes of traditional Chinese medicine].

Traditional Chinese medicines(TCMs) have certain limitations in the clinical research design in their post-marketing evaluation, so that randomized controlled programs cannot be strictly implemented in some studies, while the objective performance criteria is a reasonable external controlled research method that has been gradually recognized at home and abroad in recent years in addition to randomized controlled trial(RCT) method. It is more mature in medical devices, surgery and other research fields, but there is no relevant report in the field of post-marketing evaluation of Chinese patent medicines. In this paper, the application prospect of the objective performance criteria and the problems were discussed in the field of post-marketing evaluation of TCM. The characteristics of as TCM are more consistent with the scope of the objective performance criteria, the application of the objective performance criteria in post-marketing evaluation of Chinese patent medicines, especially in single arm research, can break through the limitations of existing conventional clinical research methods, and improve the level of evidence, with good feasibility and advantages. However, in the application process, we should pay attention to the key issues such as the selection of index, research population, follow-up period and the reference selection, to ensure the quality of research. This research group has carried out some exploration and practice in the field of post-marketing evaluation of TCM injections by using single arm combined with the objective performance criteria, hoping to establish the key technology in this field, and provide certain research and design reference for the secondary development of Chinese patent medicines.

Multiplex measurement of twelve tumor markers using a GMR multi-biomarker immunoassay biosensor.

Tumor markers play an important role in the early diagnosis and therapeutic effect monitoring of tumors. Combined detection of multiple tumor markers is a realistic way of improving the sensitivity and specificity of cancer diagnosis. To achieve this, we studied and designed a giant magneto resistance (GMR) multi-biomarker immunoassay biosensor that can simultaneously detect twelve kinds of tumor markers by integrating a GMR sensor chip, a microfluidic device, a magnetic nano-beads label, and a double antibody sandwich immunoassay method. As a proof of concept, the proposed immunosensor was utilized to detect 12 tumor markers (AFP, CEA, CYFRA21-1, NSE, SCC, PG I, PG II, CA19-9, total PSA, free PSA, free-ß-hCG, Tg) and to screen patients with lung cancer, liver cancer, digestive tract cancer, prostatic cancer, etc. The immunosensor showed excellent sensitivity, accuracy, precision and stability. Designed as a POCT device, the immunosensor also allows for portability, able to perform rapid detection wherever necessary. As a multi-analyte assay, it provides significant advantages over single-analyte tests in terms of cost per test, labor and convenience. The system's ability to simultaneously measure the concentration of multiple markers in serum samples with excellent sensitivity and accuracy allows the immunosensor to be used for early tumor diagnosis.

The elucidation of surrounding alginate gels on the pollutants degradation by entrapped nanoscale zero-valent iron.

The adsorption capacity of calcium alginate (Ca-alginate) beads was evaluated by measuring the removal of three organic compounds with different charges (malachite green, p-chlophenol and methyl orange). The diffusion was investigated in Ca-alginate hydrogel as a function of solute charge. It was found that an increased electrostatic attraction between the hydrogel and solute would improve the mobility of solute and hence enhance its adsorption efficiency. The degradation kinetics of charged pollutants by Ca-alginate with NZVI entrapped (NZVI-alginate) beads was compared to that of bare NZVI and the data followed a pseudo-first-order kinetic model. Negatively charged Ca-alginate hydrogel strongly adsorbed positively charged pollutant, which led to an enhancement in degradation rate. However, the degradation efficiency of neutral and negatively charged pollutants by NZVI-alginate was comparable with that by bare NZVI. Thus, the degradation ability of NZVI-alginate was related with the diffusion and adsorption behavior of solutes in Ca-alginate hydrogel. The experimental results showed that the free calcium ions containing in Ca-alginate had a significant impact on the adsorption and degradation behaviors of positively charged pollutant, but those of neutral and negatively charged pollutants were only moderately affected. With the dispersity of NZVI particles in beads (1:1-1:4, w/w) increasing, the degradation efficiency of malachite green was improved, whereas further increase of NZVI dispersity (1:6, w/w) brought about a depressed degradation.

[A study of the effect of interstitial (125)I seed implantation on ward nurses' hemogram and relevant protection investigations].

OBJECTIVE: To analyze the blood picture changes of the ward nurses who nurse the patients received (125)I seeds implantation. METHODS: 32 ward nurses who nurse the patients for (125)I seeds implantation were involved in the experimental group, the control group included 32 ward nurses from the medical oncology never exposed to (125)I seeds. The WBC count, hemoglobin concentration and platelet counts from the two groups were analyzed statistically, respectively. 30 patients received (125)I seeds implantation were selected randomly and γ-ray dose from the unshielded group and lead rubber cloth covering group was detected at a distance of 0, 15, 30, 50, 100 cm from patient body surface and at different angles. RESULTS: No significant statistical difference was found between the two groups of WBC count, hemoglobin concentration and platelet count (7.1 ± 1.7 vs 6.8 ± 2.0, 132 ± 11 vs 136 ± 11, 236 ± 57 vs 242 ± 61) . The measured radiation dose was close to the natural background dose (10.2-10.8 µSv/h) at a distance of 50 cm from the body surface [ (12.7 ± 4.3) µSv/h] and the same as after 0.125 mm lead equivalent (Pb) rubber cloth shield protection[ (11.2 ± 3.1) µSv/h]. CONCLUSION: Interstitial (125)I seed implantation is safe on the blood picture of the ward nurses after taking appropriate protective measures.

Adverse reactions and their management in the treatment of malignant tumors using cytokine-induced killer cells.

OBJECTIVE: To identify the causes and the corresponding management of adverse reactions during the treatment of malignant tumors using cytokine-induced killer cells. METHODS: From January 2012 to December 2012, 441 patients received a total of 1,393 autologous cytokine-induced killer cell transfusion cycles in our department. The adverse reactions after the procedures were observed (assessed using the National Cancer Institute Common Toxicity version 2.0), and targeted care and health education were delivered by nurses. RESULTS: All treatment sessions were successfully completed, and the following adverse reactions were found: grade 1/3 fever in 1.36% (19/1,393) patients; grade 2/3 fever in 0.86% (12/1,393) patients; grade 2/3 chills in 0.65% (9/1,393) patients; and grade 1/3 dizziness in 0.29% (4/1,393) patients. CONCLUSIONS: After timely intervention of the adverse reactions, all patients were treated successfully. The best timing of the CIK cell therapy for cancer patients is when the tumor burden, or the number of tumor cells, reaches the minimal level after the end of surgery, chemotherapy and radiation therapy.