RESUMO
Polymerase chain reaction (PCR) is a technique for nucleic acid amplification, which has been widely used in molecular biology. Owing to the limitations such as large size, high power consumption, and complicated operation, PCR is only used in hospitals or research institutions. To meet the requirements of portable applications, we developed a fast, battery-powered, portable device for PCR amplification and end-point detection. The device consisted of a PCR thermal control system, PCR reaction chip, and fluorescence detection system. The PCR thermal control system was formed by a thermal control chip and external drive circuits. Thin-film heaters and resistance temperature detectors (RTDs) were fabricated on the thermal control chip and were regulated with external drive circuits. The average heating rate was 32 °C/s and the average cooling rate was 7.5 °C/s. The disposable reaction chips were fabricated using a silicon substrate, silicone rubber, and quartz plate. The fluorescence detection system consisted a complementary metal-oxide-semiconductor (CMOS) camera, an LED, and mirror units. The device was driven by a 24 V Li-ion battery. We amplified HPV16E6 genomic DNA using our device and achieved satisfactory results.
Assuntos
Calefação , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase , DNA/genética , TemperaturaRESUMO
Au nanoclusters with an average size of approximately 1â nm size supported on HY zeolite exhibit a superior catalytic performance for the selective oxidation of 5-hydroxymethyl-2-furfural (HMF) into 2,5-furandicarboxylic acid (FDCA). It achieved >99â % yield of 2,5-furandicarboxylic acid in water under mild conditions (60 °C, 0.3â MPa oxygen), which is much higher than that of Au supported on metal oxides/hydroxide (TiO2 , CeO2 , and Mg(OH)2 ) and channel-type zeolites (ZSM-5 and H-MOR). Detailed characterizations, such as X-ray diffraction, transmission electron microscopy, N2 -physisorption, and H2 -temperature-programmed reduction (TPR), revealed that the Au nanoclusters are well encapsulated in the HY zeolite supercage, which is considered to restrict and avoid further growing of the Au nanoclusters into large particles. The acidic hydroxyl groups of the supercage were proven to be responsible for the formation and stabilization of the gold nanoclusters. Moreover, the interaction between the hydroxyl groups in the supercage and the Au nanoclusters leads to electronic modification of the Au nanoparticles, which is supposed to contribute to the high efficiency in the catalytic oxidation of HMF to FDCA.
Assuntos
Ácidos Dicarboxílicos/química , Ácidos Dicarboxílicos/síntese química , Furaldeído/análogos & derivados , Furanos/química , Furanos/síntese química , Ouro/química , Nanopartículas Metálicas/química , Ítrio/química , Zeolitas/química , Catálise , Furaldeído/química , Oxirredução , TemperaturaRESUMO
The purpose of this study was to evaluate the sensitivity, specificity, and predictive value of serum procalcitonin (PCT) for osteomyelitis and septic arthritis in children. PubMed, EMBase, and Cochrane Library were searched until 10 August 2021, for eligible literature focusing on PCT for the diagnosis of osteomyelitis and septic arthritis. Four articles with six studies were included in the diagnostic meta-analysis, a total of 654 children were examined for bacterial cultures in PCT, osteomyelitis, and septic arthritis. The results of diagnostic meta-analysis showed that the PCT had a sensitivity of 0.72, 95% confidence interval (CI) (0.65-0.79), specificity of 0.90, 95% CI (0.87-0.93), positive likelihood ratio (LR) of 3.87, 95% CI (2.53-5.90), negative LR of 0.39, 95% CI (0.22-0.70), and diagnostic odds ratio was 13.13, 95% CI (6.46-26.66), for the detection of osteomyelitis and septic arthritis using bacterial culture as the gold standard. Based on the summary receiver operating characteristic curve of PCT, it was found that the area under the curve of PCT was 0.88. In the evaluation of publication bias, the result of the regression line test showed that there was not publication bias (bias = 13.72; 95% CI, -1.84 to 29.28; P = 0.07). This study provided systematic review of the published literature on the diagnosis of osteomyelitis and septic arthritis in children using PCT, which may serve as a biomarker for diagnosis of osteomyelitis, but it has no direct evidences to support the diagnosis of septic arthritis. However, the specific optimal cutoff value of PCT and specific population still needed to be verified by large sample studies.
Assuntos
Artrite Infecciosa , Osteomielite , Criança , Humanos , Pró-Calcitonina , Calcitonina , Peptídeo Relacionado com Gene de Calcitonina , Precursores de Proteínas , Osteomielite/diagnóstico , Curva ROC , Artrite Infecciosa/diagnóstico , Testes Diagnósticos de RotinaRESUMO
PURPOSE: Invasive breast carcinomas (BRCAs) are highly lethal. The molecular mechanisms underlying progression of invasive BRCAs are unclear, and effective therapies are highly desired. The cancer-testis antigen CT45A1 promotes overexpression of pro-metastatic sulfatase-2 (SULF2) and breast cancer metastasis to the lungs, but its mechanisms are largely unknown. In this study, we aimed to elucidate the mechanism of CT45A1-induced SULF2 overexpression and provide evidence for targeting CT45A1 and SULF2 for breast cancer therapy. METHODS: The effect of CT45A1 on SULF2 expression was assessed using reverse transcription polymerase chain reaction and western blot. The mechanism of CT45A1-induced SULF2 gene transcription was studied using protein-DNA binding assay and a luciferase activity reporter system. The interaction between CT45A1 and SP1 proteins was assessed using immunoprecipitation and western blot. Additionally, the suppression of breast cancer cell motility by SP1 and SULF2 inhibitors was measured using cell migration and invasion assays. RESULTS: CT45A1 and SULF2 are aberrantly overexpressed in patients with BRCA; importantly, overexpression of CT45A1 is closely associated with poor prognosis. Mechanistically, gene promoter demethylation results in overexpression of both CT45A1 and SULF2. CT45A1 binds directly to the core sequence GCCCCC in the promoter region of SULF2 gene and activates the promoter. Additionally, CT45A1 interacts with the oncogenic master transcription factor SP1 to drive SULF2 gene transcription. Interestingly, SP1 and SULF2 inhibitors suppress breast cancer cell migration, invasion, and tumorigenicity. CONCLUSION: Overexpression of CT45A1 is associated with poor prognosis in patients with BRCA. CT45A1 promotes SULF2 overexpression by activating the promoter and interacting with SP1. Additionally, SP1 and SULF2 inhibitors suppress breast cancer cell migration, invasion, and tumorigenesis. Our findings provide new insight into the mechanisms of breast cancer metastasis and highlight CT45A1 and SULF2 as sensible targets for developing novel therapeutics against metastatic breast cancer.
RESUMO
High-risk human papillomavirus (HPV) testing can significantly decline the incidence and mortality of cervical cancer. Microfluidic technology provides an effective method for accurate detection of high-risk HPV by utilizing multiplex single-cell droplet polymerase chain reaction (PCR). However, current strategies are limited by low-integration microfluidic chip, complex reagent system, expensive detection equipment and time-consuming droplet identification. Here, we developed a novel multiplex droplet PCR method that directly detected high-risk HPV sequences in single cells. A multiplex microfluidic chip integrating four flow-focusing structures was designed for one-step and parallel droplet preparation. Using single-cell droplet PCR, multi-target sequences were detected simultaneously based on a monochromatic fluorescence signal. We applied machine learning to automatically identify the large populations of single-cell droplets with 97% accuracy. HPV16, 18 and 45 sequences were sensitively detected without cross-contamination in mixed CaSki and Hela cells. The approach enables rapid and reliable detection of multi-target sequences in single cells, making it powerful for investigating cellular heterogeneity related to cancer diagnosis and treatment.
Assuntos
Papillomavirus Humano , Infecções por Papillomavirus , Humanos , Células HeLa , Infecções por Papillomavirus/diagnóstico , Reação em Cadeia da Polimerase Multiplex , Aprendizado de MáquinaRESUMO
An excellent linear correlation between ln δ (OH chemical shift) and 1/T (temperature) is discovered for the first time for hydroxyl compounds. The derived slope (A) provides information as an index not only for distinguishing different types of H-bonds, but also for predicting their reactivities. This finding can be extended to other H-bond-containing molecules.
Assuntos
Éter/química , Catálise , Ligação de Hidrogênio , Hidroxilação , Temperatura , Termodinâmica , Compostos de Tungstênio/químicaRESUMO
5-Hydroxymethylfurfural (HMF) is an important biomass-derived building block, but production and sustainable utilization of HMF remain challenging due to reactions of the highly reactive functional groups of this compound. H(4)SiW(12)O(40)/MCM-41 nanospheres were developed that exhibit 84.1% selectivity to 5-ethoxymethylfurfural (EMF) when HMF conversion reaches 92.0%, during etherification of 5-hydroxymethylfurfural (HMF) with ethanol under mild conditions. The catalyst could be reused, and its activity remained unaffected over five cycles. The strong acidity of the catalyst significantly enhanced etherification. The acetalized byproducts, 5-(diethoxymethyl)-2-furanmethanol and the HMF-dimer (5,5'(oxy-bis(methylene))bis-2-furfural), can be converted into HMF and then transformed to the main product, EMF, by using this catalyst to shift the reaction equilibrium.