Quantitative assessment of disease markers using the naked eye: point-of-care testing with gas generation-based biosensor immunochromatographic strips.

BACKGROUND: Immunochromatographic strips (ICSs) are a practical tool commonly used in point-of-care testing (POCT) applications. However, ICSs that are currently available have low sensitivity and require expensive equipment for quantitative analysis. These limitations prohibit their extensive use in areas where medical resources are scarce. METHODS: We developed a novel POCT platform by integrating a gas generation biosensor with Au@Pt Core/Shell nanoparticle (Au@PtNPs)-based ICSs (G-ICSs). The resulting G-ICSs enabled the convenient and quantitative assessment of a target protein using the naked eye, without the need for auxiliary equipment or complicated computation. To assess this platform, C-reactive protein (CRP), a biomarker commonly used for the diagnosis of acute, infectious diseases was chosen as a proof-of-concept test. RESULTS: The linear detection range (LDR) of the G-ICSs for CRP was 0.05-6.25 µg/L with a limit of detection (LOD) of 0.041 µg/L. The G-ICSs had higher sensitivity and wider LDR when compared with commonly used AuNPs and fluorescent-based ICSs. When compared with results from a chemiluminescent immunoassay, G-ICS concordance rates for CRP detection in serum samples ranged from 93.72 to 110.99%. CONCLUSIONS: These results demonstrated that G-ICSs have wide applicability in family diagnosis and community medical institutions, especially in areas with poor medical resources.

[Cytotoxic effect of lappaconitine on non-small cell lung cancer in vitro and its molecular mechanism].

OBJECTIVE: To evaluate the proliferation inhibitory effect of Lappaconitine (LAP) on non-small cell lung cancer cell line A549 cells in vitro and its possible mechanism. METHODS: A549 cell was cultured with different concentrations of LAP. Cellular proliferation was determined with MTT. Cell cycle and apoptosis were detected with FCM technology. The Cyclin E1 gene expression was checked by Real-time Quantitative PCR method. RESULTS: LAP could inhibit the proliferation of A549 cells in vitro in a dose-dependent manner. LAP could induce apoptosis of A549 cell. Cell cycle was stopped at the G1 + G0 phase by LAP with FCM technology. With the increasement of LAP concentration, the ratio of G1 + G0 phase was increased and the ratio of S phase and G2 + M phase was decreased; The apoptotic rate was gradually increased, and the Cyclin E1 gene expression was down-regulated. CONCLUSION: LAP has the inhibitory effect on the growth of A549 cells, which is related to the cell cycle arrest in G0/G1 phase, apoptosis and down-regulation of Cyclin E1 gene expression.

Cell Division Cycle-Associated Genes Are Potential Immune Regulators in Nasopharyngeal Carcinoma.

BACKGROUND: Cell division cycle-associated (CDCA) gene family is essential to cell cycle regulation. Numerous studies have illuminated that dysfunction of CDCA genes may not only lead to uncontrolled cell proliferation resulting in tumorigenesis but also influence immune cell infiltration in tumors. However, the role of the CDCA gene family on the prognosis and immune infiltration in nasopharyngeal carcinoma (NPC) remains to be unclear. METHODS: SBC human ceRNA array V1.0 was used to measure mRNA expression in three pairs of NPC tissues and nasopharyngitis tissues. The expression of CDCA8 was confirmed in an IHC microarray containing 130 NPC patients. Two external GEO cohorts were enrolled for further analysis. Prognosis analysis was performed using the Kaplan-Meier method. Gene set enrichment analysis (GSEA) was applied to explore the potential mechanism of CDCA genes in NPC. The relationship between CDCA gene family and immune infiltration in NPC was evaluated using the Xcell tool. RESULTS: CDCA genes were broadly upregulated in NPC tissues compared to nasopharyngitis tissues, and high expression of CDCA3/5/8 indicated worse prognosis in NPC. Besides cell cycle pathways, we found that CDCA3/5/8 were involved in multiple immune-related pathways. Overexpression of CDCA8 was strongly associated with less infiltration of CD8+ T cells and more infiltration of CD4+ Th1 cells and was negatively correlated with immune checkpoint blockade (ICB)-related genes. CONCLUSION: CDCA gene family was upregulated in NPC, and their expressions were associated with adverse prognosis. High expression of CDCA8 was associated not only with poor prognosis, but also with less immune infiltration and downregulation of ICB-related genes in NPC.

RNA m6A reader YTHDF2 facilitates lung adenocarcinoma cell proliferation and metastasis by targeting the AXIN1/Wnt/ß-catenin signaling.

Lung adenocarcinoma (LUAD) remains a leading cause of cancer-related deaths worldwide. YTHDF2 is a reader of N6-methyladenosine (m6A) on RNA and plays a critical role in the initiation and propagation of myeloid leukemia; however, whether YTHDF2 controls the development of LUAD remains to be explored. Here, we found that YTHDF2 was significantly upregulated in LUAD compared with paracancerous normal tissues, and YTHDF2 knockdown drastically inhibited, while its overexpression promoted, cell growth, colony formation and migration of LUAD cells in vitro. In addition, YTHDF2 knockdown significantly inhibited tumorigenesis in a murine tumor xenograft model. Through the integrative analysis of RNA-seq, m6A-seq, CLIP-seq, and RIP-seq datasets, we identified a set of potential direct targets of YTHDF2 in LUAD, among which we confirmed AXIN1, which encodes a negative regulator of the Wnt/ß-catenin signaling, as a direct target of YTHDF2. YTHDF2 promoted AXIN1 mRNA decay and subsequently activated the Wnt/ß-catenin signaling. Knockout of AXIN1 sufficiently rescued the inhibitory effect of YTHDF2 depletion on lung cancer cell proliferation, colony-formation, and migration. These results revealed YTHDF2 to be a contributor of LUAD development acting through the upregulation of the AXIN1/Wnt/ß-catenin signaling, which can be a potential therapeutic target for LUAD.

Protective effect of tetrandrine on doxorubicin-induced cardiotoxicity in rats.

AIMS AND BACKGROUND: Doxorubicin (Dox) is effective in curative and adjuvant chemotherapy of malignant tumors. Cardiotoxicity is the chief toxic effect that limits the clinical use of Dox. We studied the effects of tetrandrine (Tet) on doxorubicin-induced cardiotoxicity in rats and its protective activity. MATERIALS AND METHODS: Sprague-Dawley rats were randomly divided into the following 4 groups: a control group (received only saline), Dox group (received only Dox), Tet/Dox group (received Tet plus Dox), and Tet group (received only Tet). Rats were injected intravenously with 2 mg/kg Dox once a week for 7 weeks and 50 mg/kg Tet was administered intraperitoneally weekly for 7 weeks. Measurements of cardiac contractile parameters including LSVP +dP/dt max and -dP/dt max, and assessment of electrocardiograms were carried out. Mitochondrial oxidation and phosphorylation state 3 (S3) and state 4 (S4) respiration were measured. Respiration control rate (RCR) and the ADP/O ratio were calculated. Cardiac ultrastructure was examined by electron microscopy. RESULTS: Dox induced significant cardiotoxicity in this rat model. The values of LSVP, +dP/dt max, and -dP/dt max in the Tet/Dox group increased as compared to the Dox group (P <0.05). The cardiac contraction and relaxation improved on Tet administration. Tet inhibited the prolonged QT interval on the electrocardiogram in Dox-treated rats. Compared to the Dox group, the values of S3, RCR, and ADP/O increased by more than 28%, 48%, and 27%, respectively, in the Tet/Dox group. Significant cardiac morphological protection was observed in the Tet/Dox-treated rats. CONCLUSION: Tet can improve the reduced cardiac function caused by Dox treatment and prevent Dox-induced mitochondrial impairment in rat cardiotoxicity.

[Effect of supplement energy and nourish lung herbs on drug-resistance lung cancer with mechanisms of mitochondrial and caspase signal].

OBJECTIVE: To explore the effect of supplement energy and nourish lung (SENL) herbs on the growth of drug-resistance lung cancer cells, expression of multidrug resistance-associated protein (MRP) and mechanisms of mitochondrial and Caspase signal. METHODS: Drug-resistance lung cancer cell line SW1573/2R120 was treated with SENL herbs and Adriamycin. Cellular toxicity with MTT colorimetric assay, expression of MRP protein with flow microscopy, the fluorescence intensity of mitochondria membrane potetional and Ca2+ with laser confocal microscopy, the activities of caspase-3 and caspase-8 with colorimetric assay were detected. RESULTS: Compared with control group, cellular inhibitory rate was increased obviously, expression of MRP protein was decreased, mitochondria membrane potetional and intracellular Ca2+ were decreased, the activities of caspase-3 and caspase-8 were increased in the group of SENL herbs. CONCLUSION: SENL herbs can interfere and reverse drug resistance in lung cancer by the mechanism of decreasing mitochondria membrane potetional and increasing the activities of caspase-3 and caspase-8.

Clinical relevance of the multidrug resistance­associated protein 1 gene in non­small cell lung cancer: A systematic review and meta­analysis.

The multidrug resistance­associated protein 1 (MRP1) gene has been found to be consistently overexpressed in the majority of patients with non­small cell lung cancer (NSCLC). MRP1 is known for its ability to actively decrease intracellular drug concentration, limiting the efficacy of cancer chemotherapy; however, data on the clinical relevance of MRP1 is inconclusive. In the present meta­analysis, all available published data were combined to provide an updated view on the clinicopathological relevance of MRP1 in patients with NSCLC. A systematic search was conducted to obtain relevant studies published in English, Chinese and Japanese databases. All data from patients with NSCLC who underwent testing for MRP1, by either immunohistochemistry or reverse transcription­polymerase chain reaction, were extracted and combined for further analysis. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for each selected study, with either the fixed­effects model or the random­effects model where appropriate. The quality of methodology, heterogeneities and publication bias of the included articles were also analyzed. A total of 36 clinical studies involving 3,278 patients were included in the study. It was found that the increased expression of the MRP1 gene was associated with the following subgroups of patients: Non­smokers vs. smokers (OR, 2.54; 95% CI, 1.17­5.54; P=0.019); adenocarcinoma vs. squamous cell carcinoma (OR, 1.58; 95% CI, 1.16­2.17; P=0.004); clinical stage III­IV vs. stage I­II (OR, 1.36; 95% CI, 1.11­1.66; P=0.003); lymph node metastases (OR, 1.32; 95% CI, 1.09­1.61; P=0.005); poor response to chemotherapy (OR, 0.41; 95% CI, 0.23­0.72; P=0.002) and reduced 3­year survival rate (OR, 0.40; 95% CI, 0.23­0.68; P=0.001). In conclusion, the findings from this study suggest that increase in MRP1 gene expression is associated with being a non­smoker, adenocarcinoma, advanced clinical stages and a poor response to chemotherapy in patients with NSCLC. The results from the most extensive and updated data on MRP1 support the requirement for continued investigation into the potential use of MRP1 as a biomarker/clinical indicator for NSCLC.

Reversal effect of Stephania tetrandra-containing Chinese herb formula SENL on multidrug resistance in lung cancer cell line SW1573/2R120.

This research is aimed on reversing multidrug resistance (MDR) of chemotherapy in lung cancer. According to our previous research, chemotherapeutic drugs resistance in lung cancer is mainly due to high expression of multidrug resistance-associated protein (MRP) gene and activation of caspases. The effect of stephania tetrandra-containing Chinese herbal formula, namely Supplement Energy and Nourish Lung (SENL), is effective in enhancing efficacy and reducing toxicity of chemotherapy in lung cancer. However, the underlying mechnism is largely unknown. To understand whether and how SENL herbs function on multidrug-resistance lung cancer cells, we treated a multidrug resistance lung cancer cell line, SW1573/2R120 with SENL herbs alone or together with a chemotherapeutic drug, Adriamycin (ADM). We observed that SENL herbs had a significant synergistic effect with ADM in inhibiting the growth of SW1573/2R120 cells. SENL alone and particularly together with ADM could significantly increase cell apoptotic death via mitochondria- and caspase-dependent pathway. Furthermore, we showed that SENL herbs could reverse drug resistance of lung cancer cells by decreasing MRP expression and increasing accumulation of intracellular ADM, which in turn increase the sensitivity of cancer cells to ADM. Taken together, the mechanism underlying reversal effect of drug resistance by SENL treatment was reported here and further systematical investigation on SENL herbs may lead to solve drug resistance in lung cancer chemotherapy.