5-Hydroxymethylcytosines in circulating cell-free DNA as a diagnostic biomarker for nasopharyngeal carcinoma.

OBJECTIVE: To evaluate the diagnostic value of 5-hydroxymethylcytosines (5hmC) in circulating cell-free DNA (cfDNA) for nasopharyngeal carcinoma (NPC) and to develop a diagnostic model. METHODS: Genome-wide 5hmC profiles in cfDNA from 174 NPC patients and 146 non-cancer individuals were analyzed using the 5hmC-Seal technique. A cfDNA 5hmC-based diagnostic model to identify NPC patients was developed using least absolute shrinkage and selection operator (LASSO) logistic regression, and performance was evaluated with receiver operating characteristic (ROC) curves and confusion matrices. RESULTS: The 5hmC-Seal data from patients with NPC showed a different genome-wide distribution than non-tumor samples. Our initial analysis revealed a 12-gene-based 5hmC marker panel to be an accurate diagnostic model effectively distinguishing between NPC samples and non-cancerous samples (training set: area under curve (AUC)= 0.97 [95 % CI: 0.94-0.99]; and test set: AUC= 0.93 [95 % CI: 0.88-0.98]) superior to EBV DNA testing. The diagnostic score performed well in differentiating the non-cancer subjects from early-stage NPC (training set: AUC=0.99 [95 % CI: 0.98-1]; test set: AUC=0.98 [95 % CI: 0.95-1]), and advanced-stage NPC (training set: AUC=0.96 [95 % CI: 0.93-0.99]; test set: AUC=0.93 [95 % CI: 0.88-0.98]). Notably, in EBV-negative patients, the diagnostic scores showed excellent capacity for distinguishing EBV-negative patients with NPC from non-cancer subjects in both the training set (AUC= 0.94 [95 % CI: 0.88-1]) and test set (AUC=0.91 [95 % CI: 0.81-1]). CONCLUSION: 5hmC modifications in cfDNA are promising noninvasive biomarkers for NPC, offering high sensitivity and specificity, particularly for early-stage and EBV-negative NPC.

Prognostic nutritional index and serum lactate dehydrogenase predict the prognosis of nasopharyngeal carcinoma patients who received intensity-modulated radiation therapy.

PURPOSE: This research aimed to evaluate the prognostic significance of baseline prognostic nutritional index (PNI) and lactate dehydrogenase (LDH) for the outcome of individuals diagnosed with non-metastatic nasopharyngeal carcinoma (NPC). METHODS: A retrospective analysis was conducted on data from 810 patients with non-metastatic NPC who underwent intensity-modulated radiation therapy (IMRT) with or without chemotherapy. The best cut-offs for PNI and LDH were identified by X-tile software to be 48.5 and 150, respectively. To find the independent prognostic factors for survival outcomes, univariate and multivariate regression analyses were conducted, and AUCs were used to compare their prognostic values. RESULTS: Multivariate analysis revealed that patients with PNI > 48.5 had better overall survival (OS) (HR: 0.502, P < 0.001), progression-free survival (PFS) (HR: 0.618, P < 0.001), and distant metastasis-free survival (DMFS) (HR: 0.637, P = 0.005). Higher LDH was associated with poorer OS (HR: 1.798, P < 0.001), PFS (HR: 1.671, P < 0.001), and DMFS (HR: 1.756, P < 0.001). The combination of low PNI and high LDH in non-metastatic NPC patients was correlated with poor OS (P < 0.001), PFS (P < 0.001), and DMFS (P < 0.001). The combination of PNI and LDH had the highest AUCs for predicting OS, PFS, and DMFS. CONCLUSIONS: PNI and LDH might become valuable predictors of the prognosis of non-metastatic NPC patients undergoing IMRT with or without chemotherapy. Prognostic accuracy can be enhanced by combining PNI and LDH.

Rho-kinase inhibitor, fasudil, prevents neuronal apoptosis via the Akt activation and PTEN inactivation in the ischemic penumbra of rat brain.

Recently, some studies suggested that inhibition of Rho-kinase (ROCK) prevented cerebral ischemia injury through inhibiting inflammatory reaction, increasing cerebral blood flow, modulating the neuronal actin cytoskeleton polymerization, and preventing tau hyperphosphorylation and p25/CDK5 increase. However, there is little information regarding the effects of ROCK inhibitor on the neuronal apoptosis in ischemic brain injury. In this study, we determined whether ROCK inhibitor, fasudil, inhibited ischemic neuronal apoptosis through phosphatase and tensin homolog deleted on chromosome10 (PTEN)/Akt/signal pathway in vivo. Adult male Sprague-Dawley rats were subjected to permanent middle cerebral artery occlusion. Rats received ROCK inhibitor, fasudil (10 mg/kg), at 30 min before middle cerebral artery occlusion. The infarct area, neuronal apoptosis and caspase-3 activity was significantly decreased by fasudil with improvement of neurological deterioration. However, the beneficial effects of fasudil were attenuated by the co-application of LY294002 (PI3K inhibitor). Fasudil maintained postischemic Akt activity at relatively proper level and decreased the augmentation of PTEN and ROCK activity in the penumbra area. Furthermore, fasudil inhibited attenuation of GSK-ß and Bad phosphorylation in the penumbra area. In conclusion, the findings provide another consideration that fasudil protects the brain against ischemia injury through decreasing neuronal apoptosis and reveals the link between the ROCK inhibition and the PTEN/Akt pathway.

Expression and subcellular localization of syntaxin 11 in human neutrophils.

OBJECTIVE: Syntaxin 11 mutations lead to familial hemophagocytic lymphohistiocytosis (FHL), characterized by uncontrolled hyperinflammation. This study examines the expression and subcellular localization of syntaxin 11 in human neutrophils as major inflammatory cells. MATERIALS: The materials included human peripheral blood neutrophils, HL-60 cells. METHODS: The methods used were RT-PCR, Western blot, immunocytochemistry, subcellular fractionation, HL-60 cell differentiation. RESULTS: We have found that human peripheral blood neutrophils express syntaxin 11 mRNA and protein. Syntaxin 11 was upregulated during neutrophil differentiation of HL-60 cells. Syntaxin 11, identified as a membrane-bound protein, was broadly located in the plasma membrane and granules, with a predominant location in azurophilic granules of resting human neutrophils. A secondary location of syntaxin 11 was in specific and tertiary granules, which resulted in translocation to the plasma membrane on cell activation conditions that promoted the release of these organelles. CONCLUSIONS: These data indicate that human neutrophils express syntaxin 11 and call attention to the possible involvement of neutrophils in familial hemophagocytic lymphohistiocytosis pathology.

Development a polymer-based electronic pulse diagnosis instrument for measuring and analyzing pulse wave velocity.

A novel pulse-diagnosis system was proposed in this study for measuring pulse wave velocities. In contrast with most conventional mechanical, rigid-type pulse diagnosis instruments such as pressure transducers and microactuators, a conductive elastic polymer was adopted as the sensor material. The soft and formability properties of such material enabled fabricating a flexible pulse diagnosis instrument. In addition, the flexible design was integrated with a contemporary, wrist-type pulse-wave acquisition system to ensure stable measurements. Closely related to the incidence of cardiovascular diseases, pulse wave velocity was analyzed in applications to verify the feasibility of this system. Regarding signal processing, the cun, guan, and chi pulse signals obtained through the data acquisition device were sent to the LabVIEW platform for reconstructing the pulse waveforms. Finally, the results of 20 measured samples were compared and analyzed to evaluate the level of system performance.

[Effect of jiuxinfumai injection on L-type calcium currents in single guinea pig ventricular myocytes].

OBJECTIVE: To determine the effect of 311 and 417, both active ingredients isolated from Jiuxinfumai injection (Citrus Aurantium) on L-type calcium currents (I(Ca-L)) in ventricular myocytes of guinea pigs. METHODS: Single myocytes were dissociated by enzymatic dissociation method. The whole-cell patch-clamp recording technique was used to record the change of calcium current after the administration of 311and 417. RESULTS: 311 (10, 25, 50, 100 mmol/L) increased the (I(Ca-L)) by 8.27%, 27.29%, 41.01%, and 48.74% (P < 0.05), respectively. 417 (10, 25, 50, 100 mmol/L) increased the (I(Ca-L)) by 10.05%, 30.12%, 43.05%, and 51.90% (P < 0.05), respectively. Both 311 and 417 changed the (I(Ca-L)) significantly in a concentration-dependent manner. They did not change the shape of I-V cruves. CONCLUSION: 311 and 417 can increase I(I(Ca-L)) n ventricular myocytes of guinea pigs in a dose-response manner.

Mitochondrial-derived ROS in edelfosine-induced apoptosis in yeasts and tumor cells.

AIM: To investigate whether a similar process mediates cytotoxicity of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH3, edelfosine) in both yeasts and human tumor cells. METHODS: A modified version of a previously described assay for the intracellular conversion of nitro blue tetrazolium to formazan by superoxide anion was used to measure the generation of reactive oxygen species (ROS). Apoptotic yeast cells were detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. DNA fragmentation and the generation of ROS were measured by cytofluorimetric analysis in Jurkat cells. RESULTS: Edelfosine induced apoptosis in Saccharomyces cerevisiae, as assessed by TUNEL assay. Meanwhile, edelfosine induced a time- and concentration-dependent generation of ROS in yeasts. Rotenone, an inhibitor of the mitochondrial electron transport chain, prevented ROS generation and apoptosis in response to edelfosine in S cerevisiae. alpha-Tocopherol abrogated the edelfosine-induced generation of intracellular ROS and apoptosis. Edelfosine also induced an increase of ROS in human leukemic cells that preceded apoptosis. The overexpression of Bcl-2 by gene transfer abrogated both ROS generation and apoptosis induced by edelfosine in leukemic cells. Changes in the relative mitochondrial membrane potential were detected in both yeasts and Jurkat cells. CONCLUSION: These results indicate that edelfosine induces apoptosis in yeasts in addition to human tumor cells, and this apoptotic process involves mitochondria, likely through mitochondrial-derived ROS. These data also suggest that yeasts can be used as a suitable cell model in elucidating the antitumor mechanism of action of edelfosine.

[Effect of low-dosage aspirin combined with perindopril on prostacyclin, thromboxone A2, and norepinephrine in rabbits' blood].

OBJECTIVE: To explore the interaction of low-dosage aspirin combined with angiotensin-converting enzyme (ACE) inhibitors by prostacyclin (PGI2), thromboxone A2 (TXA2) and norepinephrine (NE)) levels in rabbits' blood. METHODS: Forty healthy New Zealand rabbits were divided into four groups. Blood samples were drawn from the rabbits' heart before and after a consecutive four-week. NE was measured by high performance liquid chromatography, and PGI2 and TXA2 were measured by radioimmunoassay. RESULTS: ACE inhibitors increased PGI2 levels (P < 0.05, P < 0.01); low-dosage aspirin suppressed TXA2 productions (P < 0.05, P < 0.01) after the four-week administration. Aspirin combined with ACE inhibitors led to a significant increase in PGI2/TXA2(P < 0.01), together with a significant decrease in NE levels in the rabbits' blood (P < 0.001). CONCLUSION: Neither low-dosage aspirin nor ACE inhibitors influence NE levels alone. The ratio of PGI2 to TXA2 increased, and NE levels decreased significantly during the administration of aspirin combined with ACE inhibitors. The results suggest that there is a synergis-action between low-dosage aspirin and ACE inhibitors due to increased PGI2/TXA2 and decreased NE levels.