SPRi/SERS dual-mode biosensor based on ployA-DNA/ miRNA/AuNPs-enhanced probe sandwich structure for the detection of multiple miRNA biomarkers.

MicroRNA (miRNA) has broad application prospects in the early detection of various cancers. In this work, a SPRi/SERS dual-mode biosensor was developed on the same gold chip by AuNPs as the reinforcing medium. High throughput and sensitivity detection of three typical cervical cancer markers miRNA21, miRNA124 and miRNA143 were achieved based on the sandwich structure of polyA blocks-DNA capture probe/target miRNA/AuNPs-assistant probe or SERS nanoprobes. AuNPs greatly improved the SPR response due to mass increase and more sensitive refractive index changes. Meanwhile, due to the LSPR effect of AuNPs, the signal of SERS nanoprobe can be amplified. The miRNAs were detected in serum to verify its practicality. SPRi achieved detection of three miRNAs simultaneously. LODs were 6.3 fM, 5.3 fM and 4.6 fM, respectively, and wide dynamic response range of 500 pM-10 nM. While SERS assay ensured high sensitivity with LODs as low as 1 fM, 0.8 fM and 1.2 fM, respectively, and with the recoveries in the range of 90.0 %-100.2 %. The redundant detection signals of the two modes can provide more reliable data to prevent false positive or false negative detection, and have great application prospects in detection of cancer-related nucleic acids in early stage of disease.

Efficient interfacial self-assembled MXene/Ag NPs film nanocarriers for SERS-traceable drug delivery.

Combining the unique advantages of two-dimensional transition metal carbon/nitrogen compounds (MXene) and the excellent surface-enhanced Raman scattering (SERS) performance of noble metal materials, MXene/Ag NPs films were proposed as nanocarriers for SERS-traceable drug delivery. The films were prepared by two-step self-assembly on positively charged silicon wafers using virtue of the high evaporation of ethyl acetate, the Marangoni effect, and an oil/water/oil three-phase system. With 4-mercaptobenzoic acid (4-MBA) as the probe molecule, the SERS detection limit was 10-8 M and had shown a good linear relationship in the range of 10-8-10-3 M. Simultaneously, the film had good uniformity, repeatability, and stability. When Ti3C2Tx/Ag NPs films were used as nanocarriers, the anticancer drug doxorubicin (DOX) was loaded onto the surface through 4-MBA, and the tracking and monitoring were realized by SERS. The addition of glutathione (GSH) triggered the thiol exchange reaction, resulting in the shedding of 4-MBA from the surface of the film, which indirectly achieved the efficient release of DOX. Furthermore, the loading of DOX and the drug release effect triggered by GSH maintained a certain stability in serum, which provided a potential possibility for the subsequent loading and release of drugs by films with three-dimensional structures as scaffolds in biological therapy. Self-assembled MXene/Ag NPs film nanocarriers for SERS-traceable drug delivery and GSH-triggered high-efficiency drug release.

Structural insights into pathogenic mechanism of hypohidrotic ectodermal dysplasia caused by ectodysplasin A variants.

EDA is a tumor necrosis factor (TNF) family member, which functions together with its cognate receptor EDAR during ectodermal organ development. Mutations of EDA have long been known to cause X-linked hypohidrotic dysplasia in humans characterized by primary defects in teeth, hair and sweat glands. However, the structural information of EDA interaction with EDAR is lacking and the pathogenic mechanism of EDA variants is poorly understood. Here, we report the crystal structure of EDA C-terminal TNF homology domain bound to the N-terminal cysteine-rich domains of EDAR. Together with biochemical, cellular and mouse genetic studies, we show that different EDA mutations lead to varying degrees of ectodermal developmental defects in mice, which is consistent with the clinical observations on human patients. Our work extends the understanding of the EDA signaling mechanism, and provides important insights into the molecular pathogenesis of disease-causing EDA variants.

Case report: Novel junctional sarcoplasmic reticulum protein 1 intergenic region-anaplastic lymphoma kinase fusion in a patient with lung adenocarcinoma responds to alectinib.

Novel anaplastic lymphoma kinase (ALK) fusions are still being discovered in non-small cell lung cancer (NSCLC). Most patients with ALK+ NSCLC respond favorably to ALK tyrosine kinase inhibitors. In this case report, we identified a novel nonreciprocal ALK fusion, namely, junctional sarcoplasmic reticulum protein 1 (JSRP1) intergenic region-ALK fusion (Jintergenic: A20) via next-generation sequencing in a female patient initially diagnosed with stage IV B lung adenocarcinoma. Further examination of biopsy specimen and analysis of clinical samples by a multidisciplinary team confirmed the diagnosis of ALK+ NSCLC. At the 2- and 4-months follow-up after receiving alectinib, the patient responded rapidly, implying that alectinib had a remarkable therapeutic effect. We identified a novel JSRP1 intergenic region-ALK fusion as a carcinogenic mutation that responds to alectinib, thereby expanding the spectrum of ALK fusion partners in ALK + NSCLC. This study may help clinicians detect oncogenic mutations and provide timely treatment to patients with ALK+ NSCLC.

Two novel ectodysplasin A gene mutations and prenatal diagnosis of X-linked hypohidrotic ectodermal dysplasia.

BACKGROUND: Hypohidrotic ectodermal dysplasia (HED) is mainly caused by ectodysplasin A (EDA) gene mutation. Fetus with genetic deficiency of EDA can be prenatally corrected. This study aimed at revealing the pathogenesis of two HED families and making a prenatal diagnosis for one pregnant female carrier. DESIGNS: Genomic DNA was extracted from two HED patients and sequenced using whole exome sequencing (WES). The detected mutations were confirmed in patients and family members using Sanger sequencing. The expression of soluble ectodysplasin A1 (EDA1) protein was studied by western blot. The transcriptional activity of NF-κB pathway was tested by dual luciferase assay. The genomic DNA of fetus was extracted from shed chorion cells and EDA gene was screened through Sanger sequencing. RESULTS: We identified two novel EDA mutations: c.1136T>C (p.Phe379Ser) and c.[866G>C;868A>T] (p.[Arg289Pro;Ser290Cys]). Further examinations revealed that these two mutated EDA1 proteins showed completely impaired solubility, and the transcriptional NF-κB activation induced by these missense mutant-type EDA1 proteins was significantly reduced compared with wild-type EDA1. Furthermore, the analysis of amniotic fluid samples from a pregnant heterozygote indicated that the fetus was a c.1136T>C mutation female carrier. CONCLUSIONS: This study extended the mutation spectrum of X-linked hypohidrotic ectodermal dysplasia (XLHED) and applied prenatal diagnosis for the pregnant carrier, which can be helpful in genetic counseling, prenatal diagnosis, and intervention for the XLHED family.

Highly uniform self-assembled monolayers of silver nanospheres for the sensitive and quantitative detection of glutathione by SERS.

The homeostasis and imbalance of glutathione (GSH), an important antioxidant in organisms, are one of the key signals that reflect the health of organisms. In this paper, a novel SERS sensing platform based on Ag film@Si that self-assembled using silver nanospheres was proposed, which was used for the highly sensitive and selective detection of GSH. With the aid of an oil/water/oil three-phase system, the nano-silver film was self-assembled and finally deposited on silicon wafers. The heterobifunctional crosslinking agent N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), which contains pyridine rings and disulfide bonds, was involved in the exchange reaction between the sulfhydryl groups and disulfide bonds. With the addition of GSH, the breakage of disulfide bonds was promoted, thereby enhancing the SERS signal of SPDP. GSH can be detected sensitively by detecting the changes in the SPDP signal. The detection limit of GSH is 10 nM, and the method is still highly stable when the external environment is serum or other more complex environments.

Dihydroartemisinin Increases the Sensitivity of Photodynamic Therapy Via NF-κB/HIF-1α/VEGF Pathway in Esophageal Cancer Cell in vitro and in vivo.

BACKGROUND/AIMS: Although photodynamic therapy (PDT) can relieve esophageal obstruction and prolong survival time of patients with esophageal cancer, it can induce nuclear factor-kappa B (NF-κB) activation in many cancers, which plays a negative role in PDT. Dihydroartemisinin (DHA), the most potent artemisinin derivative, can enhance the effect of PDT on esophageal cancer cells. However, the mechanism is still unclear. METHODS: We generated stable cell lines expressing the super-repressor form of the NF-κB inhibitor IκBα and cell lines with lentivirus vector-mediated silencing of the HIF-1α gene. Esophageal xenograft tumors were created by subcutaneous injection of Eca109 cells into BALB/c nude mice. Four treatment groups were analyzed: a control group, photosensitizer alone group, light alone group, and PDT group. NF-κB expression was detected by an electrophoretic mobility shift assay, hypoxia-inducible factor α (HIF-1α) and vascular endothelial growth factor (VEGF) by real-time PCR, NF-κB, HIF-1α, and VEGF protein by western blot, and Ki-67, HIF-1α, VEGF, and NF-κB protein by immunohistochemistry. RESULTS: PDT increased NF-κB activity and the gene expression of HIF-1α and VEGF in vitro and in vivo. In contrast, the DHA groups, particularly the combined DHA and PDT treatment group, abolished the effect. The combined treatment significantly inhibited tumor growth in vitro and in vivo. NF-κB activity and HIF-1α expression were also reduced in the stable IκBα expression group, whereas the former showed no change in HIF-1α-silenced cells. CONCLUSION: DHA might increase the sensitivity of esophageal cancer cells to PDT by inhibiting the NF-κB/HIF-1α/VEGF pathway.

Anticancer Effects of Dihydroartemisinin on Human Esophageal Cancer Cells In Vivo.

Despite recent advances in chemotherapy and surgical resection, the 5-year survival rate of esophageal cancer still remains at the low level. Therefore, it is very important to discover a new agent to improve the life expectancy of patients with esophageal cancer. Dihydroartemisinin (DHA), a semisynthetic derivative of artemisinin, has recently exhibited promising anticancer activity against various cancer cells. But so far, the specific mechanism remains unclear. We have previously demonstrated that DHA reduced viability of esophageal cancer cells in a dose-dependent manner in vitro and induced cell cycle arrest and apoptosis. Here, we extended our study to further observe the efficacy of DHA on esophageal cancer cells in vivo. In the present study, for the first time, we found that DHA significantly inhibits cell proliferation in xenografted tumor compared with the control. The mechanism was that DHA induced cell apoptosis in both human esophageal cancer cell lines Eca109 and Ec9706 in vivo in a dose-dependent manner. The results suggested that DHA was a promising agent against esophageal cancer in the clinical treatment.