AMOTL2 restrains transforming growth factor-ß1-induced proliferation and extracellular matrix deposition of airway smooth muscle cells via the down-regulation of YAP1 activation.

Angiomotin-like 2 (AMOTL2) is a key modulator of signaling transduction and participates in the regulation of various cellular progresses under diverse physiological and pathological conditions. However, whether AMOTL2 participates in asthma pathogenesis has not been fully studied. In the present work, we studied the possible role and mechanism of AMOTL2 in regulating transforming growth factor-ß1 (TGF-ß1)-induced proliferation and extracellular matrix (ECM) deposition of airway smooth muscle (ASM) cells. Our results showed marked reductions in the abundance of AMOTL2 in TGF-ß1-stimulated ASM cells. Cellular functional investigations confirmed that the up-regulation of AMOTL2 dramatically decreased the proliferation and ECM deposition induced by TGF-ß1 in ASM cells. In contrast, the depletion of AMOTL2 exacerbated TGF-ß1-induced ASM cell proliferation and ECM deposition. Further research revealed that the overexpression of AMOTL2 restrained the activation of Yes-associated protein 1 (YAP1) in TGF-ß1-stimulated ASM cells. Moreover, the reactivation of YAP1 markedly reversed AMOTL2-mediated suppression of TGF-ß1-induced ASM cell proliferation and ECM deposition. Together, these findings suggest that AMOTL2 restrains TGF-ß1-induced proliferation and ECM deposition of ASM cells by down-regulating YAP1 activation.

Chronic restraint stress promotes hepatocellular carcinoma growth by mobilizing splenic myeloid cells through activating ß-adrenergic signaling.

Psychological stress promotes tumor progression and has a large impact on the immune system, particularly the spleen. The spleen plays an important role in tumor behavior. However, the role and mechanism of the spleen in hepatocellular carcinoma progression induced by stress is unclear. Here, we showed that the spleen plays a critical role in hepatocellular carcinoma growth induced by restraint stress. Our results demonstrated that restraint stress promoted hepatocellular carcinoma growth, changed the spleen structure, and redistributed splenic myeloid cells to tumor tissues. Interestingly, we found that splenectomy could inhibit hepatocellular carcinoma growth and prevent increases in myeloid cells and macrophages in tumor tissues in stressed mice. Restraint stress significantly elevated the concentration of norepinephrine in the spleen, serum and tumor tissues. Meanwhile, propranolol, an inhibitor of ß-adrenergic signaling, could inhibit hepatocellular carcinoma growth and prevent the redistribution of splenic myeloid cells induced by restraint stress, suggesting that restraint stress promotes hepatocellular carcinoma growth and redistributes splenic myeloid cells through ß-adrenergic signaling. Mechanistic studies revealed that restraint stress upregulated the expressions of CXCL2/CXCL3 in tumor tissues and changed the expression of CXCR2 in myeloid cells. SB225002, an inhibitor of CXCR2, could prevent the recruitment of myeloid cells in tumor tissues and inhibit tumor growth in stressed mice. Together, these data indicate that chronic restraint stress promotes hepatocellular carcinoma growth by mobilizing splenic myeloid cells to tumor tissues via activating ß-adrenergic signaling. The CXCR2-CXCL2/CXCL3 axis contributed to the recruitment of myeloid cells in tumor tissues induced by restraint stress.

Diagnostic performance of the Sofia® influenza A+B fluorescent immunoassay in adult outpatients in Northern Taiwan.

To evaluate the diagnostic performance of the Sofia influenza A+B fluorescent immunoassay (Sofia FIA), we performed a prospective study at the Chang Gung Memorial Hospital in Taiwan from January 2012 to December 2013. Patients who presented at out-patient clinics or the emergency department with influenza-like illness were included. Upper respiratory tract specimens were collected from oropharynx or nasopharynx. Performance of the Sofia FIA was compared to that of the Formosa One Sure Flu A/B Rapid Test. A Real-time reverse transcriptase-polymerase chain reaction assay (RT-PCR) and/or virus culture were used as reference standards. Of the 109 enrolled patients, the sensitivity, specificity, positive, and negative predictive values of the Sofia FIA to detect influenza A virus were 82%, 89%, 77%, and 89%, respectively. These parameters were 100% when the samples were from nasopharynx. The positive predictive value for influenza B virus detection was 29%. The sensitivity of the Sofia FIA for detection of influenza A virus was 93% between days 2 and 4 after onset of symptoms. For specimens with low viral loads (RT-PCR cycle threshold between 30 and 34.9), the sensitivity of The Sofia FIA was 83% (10/12). The Sofia FIA performed effectively in detecting influenza A virus infection. With nasopharyngeal samples, the performance was comparable to RT-PCR. Although influenza viral load typically decreases with time, the Sofia FIA was sensitive enough to identify influenza infecting patients presenting after several days of illness. However, a high false positive rate limits the assay's usefulness to identify influenza B virus infection.

Duodenoscope-Related Outbreak of a Carbapenem-Resistant Klebsiella pneumoniae Identified Using Advanced Molecular Diagnostics.

Background: Carbapenem-resistant Klebsiella pneumoniae infections are increasingly prevalent in North American hospitals. We describe an outbreak of carbapenem-resistant K. pneumoniae containing the blaOXA-232 gene transmitted by contaminated duodenoscopes during endoscopic retrograde cholangiopancreatography (ERCP) procedures. Methods: An outbreak investigation was performed when 9 patients with blaOXA-232 carbapenem-resistant K. pneumoniae infections were identified at a tertiary care hospital. The investigation included 2 case-control studies, review of duodenoscope reprocessing procedures, and culture of devices. Carbapenem-resistant Enterobacteriacieae (CRE) isolates were evaluated with polymerase chain reaction analysis for carbapenemase genes, and isolates with the blaOXA-232 gene were subjected to whole-genome sequencing and chromosome single-nucleotide polymorphism analysis. On recognition of ERCP as a key risk factor for infection, targeted patient notification and CRE screening cultures were performed. Results: Molecular testing ultimately identified 17 patients with blaOxa-232 carbapenem-resistant K. pneumoniae isolates, including 9 with infections, 7 asymptomatic carriers who had undergone ERCP, and 1 additional patient who had been hospitalized in India and was probably the initial carrier. Two case-control studies established a point-source outbreak associated with 2 specific duodenoscopes. A field investigation of the use, reprocessing, and storage of deuodenoscopes did not identify deviations from US Food and Drug Administration or manufacturer recommendations for reprocessing. Conclusions: This outbreak demonstrated the previously underappreciated potential for duodenoscopes to transmit disease, even after undergoing high-level disinfection according to manufacturers' guidelines.

Identification of curcumin-inhibited extracellular matrix receptors in non-small cell lung cancer A549 cells by RNA sequencing.

Curcumin is a potent anti-cancer drug in several types of human cancers. Despite of several preclinical and clinical studies of curcumin, the precise mechanism of curcumin in cancer prevention has remained unclear. In our study, we for the first time investigated whole transcriptome alteration in A549 non-small cell lung cancer (NSCLC) cell lines after treatment with curcumin using RNA sequencing. We found that lots of genes and signaling pathways were significantly altered after curcumin treatment in A549 cells. With bioinformatics approaches (gene ontology, Kyoto Encyclopedia of Genes and Genomes, and STRING), we found that those curcumin altered genes were not only the genes that induce cell death but also those extracellular matrix receptors and mitogen-activated protein kinase signaling pathway genes which regulate cell migration and proliferation. Among those significantly altered genes, eight genes ( COL1A1, COL4A1, COL5A1, LAMA5, ITGA3, ITGA2B, DDIT3, and DUSP1) were further examined by quantitative reverse transcription polymerase chain reaction and western blot analysis in four non-small cell lung cancer cell lines. Both in cell lines and in mouse model, the extracellular matrix receptors including the integrin ( ITGA3 and ITGA2B), collagen ( COL5A1), and laminin ( LAMA5) were significantly inhibited by curcumin at messenger RNA and protein levels. Functional studies confirmed that curcumin not only induced A549 cell death but also repressed cell proliferation and migration by regulating extracellular matrix receptors. Collectively, our study suggests that curcumin may be used as a promising drug candidate for intervening lung cancer in future studies.

A Potent Virus-Specific Antibody-Secreting Cell Response to Acute Enterovirus 71 Infection in Children.

BACKGROUND: Enterovirus 71 (EV71) remains a leading pathogen for acute infectious diseases in children, especially in Asia. The cellular basis for establishing a virus-specific antibody response to acute EV71 infections is unclear in children. METHODS: We studied the magnitude of virus-specific antibody-secreting B cells (ASCs) and its relationship with serological response, clinical parameters, and virological parameters among children with laboratory-confirmed EV71 infection. RESULTS: A potent EV71 genogroup B- and virus-specific ASC response was detected in the first week of illness among genotype B5 EV71-infected children. The cross-reactive EV71-specific ASC response to genogroup C viral antigens composed about 10% of the response. The EV71-specific ASC response in children aged ≥3 years produced immunoglobulin G predominantly, but immunoglobulin M was predominant in younger children. Proliferation marker was expressed by the majority of circulating ASCs in the acute phase of EV71 infection. Virus-specific ASC responses significantly correlated with throat viral load, fever duration, and serological genogroup-specific neutralization titer. CONCLUSIONS: The presence of a virus-specific ASC response serves an early cellular marker of an EV71-specific antibody response. Further detailed study of EV71-specific ASCs at the monoclonal level is crucial to delineate the specificity and function of antibody immunity in children.

The effects of subclinical hypothyroidism on serum lipid level and TLR4 expression of monocyte in peripheral blood of rats.

OBJECTIVE: To observe effect of subclinical hypothyroidism (SCH) on serum lipid level and expression of toll-like receptor 4 (TLR4) in rats' peripheral blood mononuclear cells (PBMC). METHODS: Fifty Wistar female rats were divided into three groups: normal control (NC group; n=10), sham group (n=10), and L-T-4 (L-thyroxine) group (n=30, with thyroidectomy, fed with rich-calcium water after operation. 5 weeks later, abdominal subcutaneous injection of L-T-4: 0.95 µg/100g/d). 8 weeks later, the rats were killed then the peripheral blood was collected to determine the levels of serum thyroid-stimulating hormone (TSH), total thyroid hormone (TT4), total cholesterol (TC) and low density lipoprotein cholesterin (LDL-C). Rats in L-T-4 group were divided into normal lipid (NL) group) and high lipid (HL) group) according to lipid value of NC group. Monocytes were separated from blood to determine TLR4 expression by flow cytometry. RESULTS: In NL and HL groups TSH were higher than in NC and Sham groups (p<0.05). TT4 have no significant differences (p>0.05). TLR4, TLR4 mRNA, NF-κB (p65) were increased (p<0.05). TNF-α, IL-6 and IL-1ß were higher than in NC and sham groups (p<0.01). There were no significant differences of TLR4, TLR4 mRNA, NF-κB (p65), TNF-α, IL-6 and IL-1ß expression between NL and HL groups (p>0.05). CONCLUSION: TLR4, TLR4 mRNA, NF-κB (p65) of PBMC and TNF-α, IL-6, IL-1ß expression in serum were all increased in SCH rats, which was not related to serum dyslipidemia.

Atorvastatin decreases Toll-like receptor 4 expression and downstream signaling in human monocytic leukemia cells.

PURPOSE: We aim to evaluate the effects of atorvastatin on the expression of Toll-like receptor (TLR4) protein and mRNA, and to explore their effects on TLR4-dependent downstream signaling in human monocytic leukemia (THP-1) cells. METHODS: TLR4 protein and mRNA expression, levels of NF-κB protein, and expression of TNF-α, IL-6, and IL-1ß in lipopolysaccharide-induced THP-1 cells after incubation with different concentrations of atorvastatin (0.1, 1, 10, 20µM) were quantified via flow-cytometry, quantitative RT-PCR, western blotting, and ELSIA kits. RESULTS: Atorvastatin incubation resulted in significant decreases in the levels of TLR4 protein and mRNA, NF-κB expression, and levels of TNF-α, IL-6, and IL-1ß in LPS-induced THP-1 cells (P<0.01). However, compared with the untreated control, the expression of these were significantly increased (P<0.01). CONCLUSIONS: Atorvastatin could inhibit the TLR4 expression and TLR4-dependent downstream signaling in THP-1 cells. These observations imply that the interactions with innate immunity may serve as one of the pleiotropic mechanisms of atorvastatin.

Top-Emitting Active-Matrix Quantum Dot Light-Emitting Diode Array with Optical Microcavity for Micro QLED Display.

An electroluminescent quantum-dot light-emitting diode (QLED) device and a micro QLED device array with a top-emitting structure were demonstrated in this study. The QLED device was fabricated in the normal structure of [ITO/Ag/ITO anode]/PEDOT:PSS/PVK/QDs/[ZnO nanoparticles]/Ag/MoO3, in which the semi-transparent MoO3-capped Ag cathode and the reflective ITO/metal/ITO (IMI) anode were designed to form an optical microcavity. Compared with conventional bottom-emitting QLED, the microcavity-based top-emitting QLED possessed enhanced optical properties, e.g., ~500% luminance, ~300% current efficiency, and a narrower bandwidth. A 1.49 inch micro QLED panel with 86,400 top-emitting QLED devices in two different sizes (17 × 78 µm2 and 74 × 40.5 µm2) on a low-temperature polysilicon (LTPS) backplane was also fabricated, demonstrating the top-emitting QLED with microcavity as a promising structure in future micro display applications.

Metabolic Dysfunction-associated Fatty Liver Disease is Associated with Greater Impairment of Lung Function than Nonalcoholic Fatty Liver Disease.

Background and Aims: We compared lung function parameters in nonalcoholic fatty liver disease (NAFLD) and metabolic dysfunction-associated fatty liver disease (MAFLD), and examined the association between lung function parameters and fibrosis severity in MAFLD. Methods: In this cross-sectional study, we randomly recruited 2,543 middle-aged individuals from 25 communities across four cities in China during 2016 and 2020. All participants received a health check-up including measurement of anthropometric parameters, biochemical variables, liver ultrasonography, and spirometry. The severity of liver disease was assessed by the fibrosis (FIB)-4 score. Results: The prevalence of MAFLD was 20.4% (n=519) and that of NAFLD was 18.4% (n=469). After adjusting for age, sex, adiposity measures, smoking status, and significant alcohol intake, subjects with MAFLD had a significantly lower predicted forced vital capacity (FVC, 88.27±17.60% vs. 90.82±16.85%, p<0.05) and lower 1 s forced expiratory volume (FEV1, 79.89±17.34 vs. 83.02±16.66%, p<0.05) than those with NAFLD. MAFLD with an increased FIB-4 score was significantly associated with decreased lung function. For each 1-point increase in FIB-4, FVC was diminished by 0.507 (95% CI: -0.840, -0.173, p=0.003), and FEV1 was diminished by 0.439 (95% CI: -0.739, -0.140, p=0.004). The results remained unchanged when the statistical analyses was performed separately for men and women. Conclusions: MAFLD was significantly associated with a greater impairment of lung function parameters than NAFLD.

Molecular Epidemiology of Cytomegalovirus UL97 and UL54 variants in Taiwan.

BACKGROUND: The antiviral resistance of cytomegalovirus (CMV) infections is associated with mutations in the CMV UL54 and UL97 gene regions and is a serious problem in immunocompromised patients. However, the molecular epidemiology of UL54 and UL97 in Taiwan is unclear. METHODS: We conducted a retrospective study of patients with CMV infections between January and December 2016 in two tertiary hospitals, one regional hospital in Taiwan. CMV DNAemia was confirmed by elevated CMV DNA titers. Then the regions of the UL54 and UL97 mutations were amplified by PCR and sequenced. RESULTS: Of 729 patients with CMV syndrome, 112 CMV DNAemia patients were enrolled. Twelve novel variants in UL54 (P342S, S384F, K434R, S673F, T754M, R778H, C814S, M827I, G878E, S880L, E888K, and S976N) and one novel variant in UL97 (M615T) were discovered. UL97 antiviral resistance mutations (L595S, M460I, and M460V) were found in four patients (3.6%). In the drug resistance strains, the mutation events occurred after 83-150 days of therapy, and drug resistance was also observed in these patients. The following high frequency variants were observed: D605E in UL97 and A885T, N898D, V355A, N685S, and A688V in UL54. CONCLUSION: The results demonstrate that the positive rate of CMV DNAemia was 15.3% (112/729) among the patients with clinical CMV infection symptoms. The proportion of antiviral resistance CMV strains within CMV DNAemia patients was 3.6%. With the information of polymorphism incidence in the UL54 and UL97 patients from our study, determination of the genetic profile of UL54 and UL97 among immunocompromised populations with refractory CMV infection is recommended.

Diagnosis of Pneumocystis pneumonia by real-time PCR in patients with various underlying diseases.

BACKGROUND: Pneumocystis pneumonia (PCP) is a disease caused by the opportunistic infection of the fungus Pneumocystis jirovecii. Several PCR methods have been developed to aid in the diagnosis of PCP. In this study, we evaluated the performance of a real-time PCR in the diagnosis of PCP, in patients with various underlying diseases. METHODS: Ninety-seven BAL samples and 94 sputum samples from 191 patients were used in the study. Patients were classified as PCP (121 patients) or non-PCP (70 patients) based on their clinical and radiological presentations. RESULTS: Real time PCR amplified the P. jirovecii mitochondrial large-subunit rRNA gene with a detection limit of 68 copies of DNA per reaction. Non-PCP pathogens including 32 different fungi and bacteria were also evaluated. Overall, 71.9% of the samples from PCP patients and 14.5% of those from non-PCP patients were positive for the PCR test with a CT value of the real-time PCR below 45. The main underlying diseases of the patients were hematological or solid malignancies (47.1%) and HIV infection (8.9%). The CT values of the test were significantly lower in BAL samples from PCP patients than those from non-PCP patients (p = 0.024). No non-PCP patient had a CT value below 30, whereas samples from 24.8% of PCP patients with underlying diseases had a CT value below 30. CONCLUSION: Since false positive PCR results were obtained, perhaps due to colonization, we suggest that the diagnosis of PCP should be based on a combination of clinical symptoms, underlying diseases, and PCR results.

Association of genetic and immuno-characteristics with clinical outcomes in patients with RET-rearranged non-small cell lung cancer: a retrospective multicenter study.

BACKGROUND: Rearranged during transfection (RET) has been proven to be a tumorigenic target in non-small cell lung cancers (NSCLCs). In RET-rearranged NSCLCs, molecular features and their impact on prognosis were not well illustrated, and the activity of mainstay therapeutics has not currently been well compared. METHODS: Patients diagnosed with NSCLCs with RET rearrangements were analyzed for concomitant mutations, tumor mutation burden (TMB), PD-L1 expression, T cell receptor repertoire and clinical outcomes with chemotherapy, immune checkpoint inhibitors (ICIs), and multikinase inhibitors (MKIs). RESULTS: Among 129 patients with RET-rearranged NSCLC who were analyzed, 41.1% (53/129) had co-occurring genetic alterations by next-generation sequencing, and concomitant TP53 mutation appeared most frequently (20/53, 37.7%). Patients with concurrent TP53 mutation (n = 15) had shorter overall survival than those without (n = 30; median, 18.4 months [95% CI, 8.6-39.1] vs 24.8 months [95% CI, 11.7-52.8]; P < 0.05). Patients with lower peripheral blood TCR diversity (n = 5) had superior overall survival compared with those with higher diversity (n = 6; median, 18.4 months [95% CI, 16.9-19.9] vs 4.8 months [95% CI, 4.5-5.3]; P = 0.035). An association with overall survival was not observed for PD-L1 expression nor for tumor mutation burden level. Median progression-free survival was not significantly different across chemotherapy, ICIs, and MKIs (median, 3.5 vs 2.5 vs 3.8 months). For patients treated with ICIs, the disease control rate was 60% (6/10) and the objective response rate was 20% (2/10). CONCLUSIONS: RET-rearranged lung cancers can be heterogeneous in terms of concomitant genetic alterations. Patients with concurrent TP53 mutation or high peripheral blood TCR repertoire diversity have relatively inferior overall survival in this series. Outcomes with traditional systemic therapies in general are suboptimal.

[Application of liquid chip-mass spectrometry technology for screening serum biomarker proteins in lung squamous cell carcinoma].

OBJECTIVE: To screen the serum biomarker proteins of lung squamous cell carcinoma (SCCs) by liquid chip-mass spectrometry technology. METHODS: All serum samples, including 34 SCCs, 46 benign lung diseases (BLDs) and 44 healthy individuals, were analyzed by CLINPROT system in order to study the serum protein expression profiles. Then the discriminatory proteins were detected by FlexAnalysis 3.0 software. Biomarkers were identified by liquid chromatography-tandem mass spectrometry (LCMS/MS). RESULTS: Comparing the differential serum expression proteins between SCCs and healthy individuals, and SCCs and BLDs respectively. Ninety-six differential protein peaks [mass-to-charge ration (M/Z) between 800 and 10 000] were found between SCCs and healthy individuals. In these protein peaks, the expression of protein peaks at 4054.13 M/Z and 4267.46 M/Z had the largest difference between them. The two protein peaks could accurately separate SCCs from healthy individuals by the frame of axes. Similarly, 99 differential protein peaks were automatically detected between SCCs and BLDs. In these protein peaks, the expression of protein peaks at 5065.27 M/Z and 4054.02 M/Z had the largest difference between them. The two protein peaks could accurately separate SCCs from BLDs by the frame of axes. Identified by LC-MS/MS, 1778 M/Z and 1865 M/Z might be assayed jointly and corresponded to complements C3 fragment or C3f precursor. CONCLUSIONS: Differential protein expressions existed between SCCs versus healthy individuals and SCCs versus BLD patients. It is feasible to screen the diagnostic serum biomarkers of SCC with a high sensitivity and specificity by using CLINPROT system.

[The value of (18)F-FDG PET/CT in differential diagnosis of benign and malignant pulmonary lesions: a Meta-analysis.].

OBJECTIVE: Using Meta analysis to evaluate the value of (18)F-FDG PET/CT ((18)fluorine-fluorodeoxyglucose Positron emission tomography/computed tomography) in differentiating between benign and malignant pulmonary lesions. METHODS: Relevant documentations from PubMed and other 5 databases from 1980 to 2008 were searched, and the eligible literatures according to the inclusive criteria were selected. The statistical information and quality of science were assessed and classified. The data were analyzed using Meta-Disc1.4 software. The diagnostic value of PET/CT in distinguishing benign from malignant pulmonary lesions was evaluated by the pooled sensitivity, specificity, the likelihood ratio (LR) and summary receiver operating characteristic curve (SROC curve) statistical indicators. RESULTS: Seven literatures were collected including 5 in English and 2 in Chinese, and 795 cases were included in the study. Heterogeneity test showed that the homogeneity of the study was good. By using deterministic models to analyze the data, the value of the weighted sensitivity was 95% (93% - 97%), the specificity was 77% (71% - 82%), the positive likelihood ratio was 4.12, negative likelihood ratio was 0.08, and the SROC area under the curve (area under curve, AUC) was 94%. CONCLUSION: PET/CT is of high diagnostic value in differentiation between benign and malignant lung lesions, but large sized, multicenter, prospective studies are needed to assess its clinical value more accurately.

[Application of laser capture microdissection and surface-enhanced laser desorption ionization time-of-flight mass spectrometry to screen biomarkers for early diagnosis of lung adenocarcinoma].

OBJECTIVE: To improve diagnostic methods to screen biomarkers for early diagnosis in lung adenocarcinoma by employing laser capture microdissection (LCM) and surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and support vector machine (SVM). METHODS: Frozen sections of thickness 8 microm were made using 6 cases of fresh lung cancer tissues and 4 cases of matched normal lung tissues. The sections were stained by improved HE solution. The homogeneous adenocarcinoma cells and normal cells were collected by LCM in each sample, and then SELDI profiles based on PBS II+SELDI-TOF-MS (IMAC protein chip) were analyzed using SVM. RESULTS: High quality cell samples were obtained by LCM quickly and precisely from normal specimens and diseased tissues without interstitium, inflammation and necrosis. Eighty four differential protein peaks were found. Top ten of them were identified as candidate biomarkers; six proteins were significantly weakly expressed in lung cancer tissue compared to normal tissues, but the other four protein were over-expressed (P < 0.05). Every candidate biomarker has undergone the blind-cross-test. Each of them can separate the lung cancer from normal samples with a sensitivity of 100% and a specificity of 100%. The 3191 m/z was considered as disease marker of lung adenocarcinoma. CONCLUSION: The method combined LCM with SELDI-TOF-MS may be able to screen potential biomarkers to distinguish lung cancer from healthy tissue with high sensitivity and specificity, which could improve early diagnosis for lung cancer.

Fluorescent Double-Stranded DNA-Templated Copper Nanoprobes for Rapid Diagnosis of Tuberculosis.

In this work, we investigate highly sensitive fluorescent Cu nanoparticles for use as rapid and specific nucleic acid amplification nanoprobes (NPs) for the diagnosis of tuberculosis. After applying polymerase chain reaction (PCR) to a tuberculosis (TB) sample, we demonstrate that the presence of the targeted IS6110 DNA sequence of TB can be easily and directly detected through the in situ formation of DNA-templated fluorescent Cu NPs and subsequently quantified using only a smartphone. Compared to traditional DNA analysis, this sensing platform does not require purification steps and eliminates the need for electrophoresis to confirm the PCR results. After optimization, this dsDNA-Cu NP-PCR method has the ability to analyze clinical TB nucleic acid samples at a detection limit of 5 fg/µL, and the fluorescent signal can be distinguished in only ∼3 min after the DNA has been amplified. Moreover, with the combination of smartphone-assisted imaging analysis, we can further reduce the instrument size/cost and enhance the portability. In this manner, we are able to eliminate the need for a fluorescent spectrophotometer to measure the clinical sample. These results demonstrate this platform's practical applicability, combining a smartphone and on-site analysis while retaining the detection performance, making it suitable for clinical DNA applications in resource-limited regions of the world.

Trends and Patterns of Disparities in Burden of Lung Cancer in the United States, 1974-2015.

Background: Although lung cancer incidence and mortality have been declining since the 1990s, the extent to which such progress has been made is unequal across population segments. Updated epidemiologic data on trends and patterns of disparities are lacking. Methods: Data on lung cancer cases and deaths during 1974 to 2015 were extracted from the Surveillance, Epidemiology, and End Results program. Age-standardized lung cancer incidence and mortality and their annual percent changes were calculated by histologic types, demographic variables, and tumor characteristics. Results: Lung cancer incidence decreased since 1990 (1990 to 2007: annual percent change, -0.9 [95% CI, -1.0%, -0.8%]; 2007 to 2015: -2.6 [-2.9%, -2.2%]). Among adults aged between 20 and 39 years, a higher incidence was observed among females during 1995 to 2011, after which a faster decline in female lung cancer incidence (males: -2.5% [-2.8%, -2.2%]; females: -3.1% [-4.7%, -1.5%]) resulted in a lower incidence among females. The white population had a higher incidence than the Black population for small cell carcinoma since 1987. Black females were the only group whose adenocarcinoma incidence plateaued since 2012 (-5.0% [-13.0%, 3.7%]). A higher incidence for squamous cell carcinoma was observed among Black males and females than among white males and females during 1974 to 2015. After circa 2005, octogenarians and older patients constituted the group with the highest lung cancer incidence. Incidence for localized and AJCC/TNM stage I lung cancer among octogenarians and older patients plateaued since 2009, while mortality continued to rise (localized: 1.4% [0.6%, 2.1%]; stage I: 6.7% [4.5%, 9.0%]). Conclusions: Lung cancer disparities prevail across population segments. Our findings inform effective approaches to eliminate lung cancer disparities by targeting at-risk populations.

[Use of laser capture microdissection and surface-enhanced laser desorption ionization time-of-flight mass spectrometry to screen differential proteins in lung adenocarcinoma and lung squamous carcinoma].

OBJECTIVE: To screen biomarkers for classification in lung adenocarcinoma and lung squamous carcinoma by using laser capture microdissection (LCM) and surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and support vector machine (SVM). METHODS: Six specimens of lung adenocarcinoma tissues and seven specimens of lung squamous carcinoma tissues obtained during operation were made into frozen sections and stained by improved H-E solution. About 1.2 x 10(5) of homogeneous adenocarcinoma cells and 1.4 x 10(5) of homogeneous lung squamous carcinoma cells were collected using LCM. Then SELDI profiles based on PBS II(+)SELDI-TOF-MS (IMAC protein chip) and the data were analyzed by support vector machine (SVM). RESULTS: Eighty seven differential protein peaks were found and top ten of them were identified as candidate biomarkers. The expression levels of 6 proteins among them with the molecular weights of 3333, 3592, 3848, 5036, 5191, and 5211 respectively in the lung squamous cancer tissues were weaker than those in the adenocarcinoma tissues, and the expression levels of 4 proteins with the molecular weights of 2505, 4004, 4847, and 11 412 in the lung squamous carcinoma tissues were stronger than those in the adenocarcinoma tissues. The expression of the protein with the molecular weight of 4847 in the squamous cancer was significantly stronger than that in the adenocarcinoma (p + 0.032). A discriminatory pattern consisting of 3 proteins with the molecular weights of 4847, 11 412, and 3592 was established with a sensitivity of 100% and a specificity of 100% respectively in separating adenocarcinoma from squamous carcinoma. CONCLUSION: There is a difference in protein component between adenocarcinoma and squamous carcinoma. LCM combined with SELDI-TOF-MS help screen a biomarker pattern to distinguish lung adenocarcinoma from lung squamous carcinoma with high sensitivity and specificity.

Detection of common diarrhea-causing pathogens in Northern Taiwan by multiplex polymerase chain reaction.

Conventional methods for identifying gastroenteritis pathogens are time consuming, more likely to result in a false-negative, rely on personnel with diagnostic expertise, and are dependent on the specimen status. Alternatively, molecular diagnostic methods permit the rapid, simultaneous detection of multiple pathogens with high sensitivity and specificity. The present study compared conventional methods with the Luminex xTAG Gastrointestinal Pathogen Panel (xTAG GPP) for the diagnosis of infectious gastroenteritis in northern Taiwan. From July 2015 to April 2016, 217 clinical fecal samples were collected from patients with suspected infectious gastroenteritis. All specimens were tested using conventional diagnostic techniques following physicians' orders as well as with the xTAG GPP. The multiplex polymerase chain reaction (PCR) approach detected significantly more positive samples with bacterial, viral, and/or parasitic infections as compared to conventional analysis (55.8% vs 40.1%, respectively; P < .001). Moreover, multiplex PCR could detect Escherichia coli O157, enterotoxigenic E coli, Shiga-like toxin-producing E coli, Cryptosporidium, and Giardia, which were undetectable by conventional methods. Furthermore, 48 pathogens in 23 patients (10.6%) with coinfections were identified only using the multiplex PCR approach. Of which, 82.6% were from pediatric patients. Because the detection rates using multiplex PCR are higher than conventional methods, and some pediatric pathogens could only be detected by multiplex PCR, this approach may be useful in rapidly diagnosing diarrheal disease in children and facilitating treatment initiation. Further studies are necessary to determine if multiplex PCR improves patient outcomes and reduces costs.