Photogenerated-hole-induced rapid elimination of solid tumors by the supramolecular porphyrin photocatalyst.

The rapid, complete, targeted and safe treatment for tumors remains a key issue in cancer therapy. A novel treatment of solid tumors by supramolecular photocatalyst Nano-SA-TCPP with the irradiation of 600-700 nm wavelength is established. Solid tumors (100 mm3) can be eliminated within 10 min. The 50-day mouse survival rate was increased from 0% to 100% after the photocatalytic therapy. The breakthrough was owing to the cell membrane rupture and the cytoplasmic loss caused by photogenerated holes inside cancer cells. The porphyrin-based photocatalysts can be internalized in a targeted manner by cancer cells due to the size selection effect, without entering the normal cells. The therapy has no toxicity or side effects for normal cells and organisms. Moreover, the photocatalytic therapy is effective for a variety of cancer cell lines. Because of its high efficiency, safety and universality, the photocatalytic therapy provides us with a new lancet to conquer the tumor.

In Situ Identification and Spatial Mapping of Microplastic Standards in Paramecia by Secondary-Ion Mass Spectrometry Imaging.

Microplastics (MPs) are universally present in the ecosystem and pose great threats to the environment and living organisms. Research studies have shown that small MPs (<50 µm in diameter) are especially toxic and account for more than half of all MPs collected in the Atlantic Ocean. Nevertheless, current methods for the detection and analysis of MPs are incapable of achieving rapid and in situ analysis of small MPs in the biota to ultimately enable the study of their biological effects. In this work, we report a method that allows rapid in situ identification and spatial mapping of small MPs directly from paramecia with high accuracy by acquiring chemical composition information using secondary-ion mass spectrometry (SIMS) imaging. Specifically, six types of common MPs (polymethyl methacrylate, polyvinyl chloride, polypropylene, polyethylene terephthalate, polyglycidyl methacrylate, and polyamide 6) with a diameter of 1-50 µm were simultaneously imaged with high chemical specificity at a spatial resolution of 700 nm. In situ spatial mapping of a group of MPs ingested by paramecia was performed using SIMS fragments specific to the plastic composition with no sample pretreatment, revealing the aggregation of MPs in paramecia after ingestion. Compared with existing methods, one additional advantage of the developed method is that the MPs and the organism can be analyzed in the same experimental workflow to record their fingerprint spectra, acquiring biochemical information to evaluate MP fate, toxicity, and the MP-biota interaction.

Dynamic Monitoring of Phase-Separated Biomolecular Condensates by Photoluminescence Lifetime Imaging.

The formation of biomolecular condensates is driven by liquid-liquid phase separation, which is prevalent in cells to govern crucial cellular functions. However, understanding the properties of phase-separated condensates remains very challenging for the lack of suitable techniques. Here, we report a photoluminescence lifetime imaging method for real-time monitoring of phase-separated condensates, both in vitro and in living cells, using a microsecond-scale photoluminescence lifetime probe based on iridium complex. The probe has a large Stokes shift, excellent cell permeability, and minimal cell autofluorescence interference. With this method, the dynamic process of phase separation of fused in sarcoma protein has been well explored, showing high spatiotemporal resolution and high throughput. Beginning with initial formation, the protein droplets get bigger and more viscous, and then a final maturation to solidified aggregates has been characterized. This study paves the path for a deeper understanding of the properties of phase-separated biomolecular condensates.

Thickness-dependent Young's modulus of polycrystalline α-PbO nanosheets.

Litharge, in two dimensional (2D) nanostructure form, has recently ignited considerable theoretical interest due to its excellent photoelectric and magnetic properties. However, the lack of an efficient synthesis method hinders its development. Here, we provide an interfacial solvothermal strategy for controllably synthesizing ultrathin hexagonal polycrystalline α-PbO nanosheets in micrometer scale. This strategy can also be utilized for the synthesis of other 2D materials. Experimental atomic force microscope nanoindentation measurements reveal the relationship between the thickness of polycrystalline α-PbO nanosheets and the corresponding Young's modulus, expressed as E = E0 + Kt -1. First-principles calculation supports the result and ascribes the cause to interlayer sliding from particular weak interlayer interactions. Additionally, the enhanced mechanical strength of the polycrystalline structure compared to its single-crystal counterpart is attributed to the alternate arrangement of grain-boundaries effects. The summative formula may be extended to other 2D materials with weak interlayer interactions, which has the potential to provide guidance for constructing flexible devices.

An iridium complex-based probe for photoluminescence lifetime imaging of human carboxylesterase 2 in living cells.

A novel photoluminescence lifetime probe (Ir-TB) has been developed for the detection and imaging of hCE2 in living cells. A large lifetime increase by around 300 ns after the enzymatic reaction makes it an ideal tool to distinguish hCE2-hydrolyzed probes from those non-hydrolyzed ones via PLIM for the first time.

Photoluminescence Lifetime Imaging of Synthesized Proteins in Living Cells Using an Iridium-Alkyne Probe.

Designing probes for real-time imaging of dynamic processes in living cells is a continuous challenge. Herein, a novel near-infrared (NIR) photoluminescence probe having a long lifetime was exploited for photoluminescence lifetime imaging (PLIM) using an iridium-alkyne complex. This probe offers the benefits of deep-red to NIR emission, a long Stokes shift, excellent cell penetration, low cytotoxicity, and good resistance to photobleaching. This example is the first PLIM probe applicable to the click reaction of copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) with remarkable lifetime shifts of 414 ns, before and after click reaction. The approach fully eliminates the background interference and distinguishes the reacted probes from the unreacted probes, thus enabling the wash-free imaging of the newly synthesized proteins within single living cells. Based on the unique properties of the iridium complexes, it is anticipated to have applications for imaging other processes within living cells.

Wnt10B is critical for the progression of gastric cancer.

The family of Wnt proteins have been implicated in embryogenesis by regulation of cell fate and pattern formation, and also in human carcinogenesis. Wnt10B was previously shown to be involved in breast cancer development. The present study assessed the association of Wnt10B expression in human gastric cancer tissue specimens with clinicopathological data from these patients. Wnt10B expression in the regulation of gastric cancer cell proliferation and migration capacity in vitro was then investigated. The data revealed that Wnt10B mRNA and protein were upregulated in gastric cancer tissue samples and the upregulated Wnt10B mRNA was associated with gastric cancer metastasizing to lymph nodes. Knockdown of Wnt10B expression reduced gastric cancer cell proliferation and migration, as well as expression of a cell proliferation marker Ki67. Knockdown of Wnt10B expression inhibited tumor cell epithelial-mesenchymal transition by upregulation of E-cadherin and downregulation of N-cadherin. In addition, Wnt10B knockdown also suppressed tumor cell stemness by downregulation of octamer-binding transcription factor 4 and Nanog expression. The present data indicated that Wnt10B expression performs an important role in gastric cancer progression in vitro. Therefore, targeting of Wnt10B expression or activity may be investigated as a possible strategy for the control of gastric cancer.

Acinetobacter baumannii Infection and IL-17 Mediated Immunity.

Acinetobacter baumannii is a significant cause of severe hospital-acquired infections with a recent rise in multidrug-resistant infections involving traumatic wounds of military personnel. The interleukin-17 (IL-17) pathway is essential for neutrophil recruitment in response to a variety of pathogens, while the control of A. baumannii infection is known to be dependent on neutrophils. This suggests that IL-17 may play an important role in A. baumannii infection; however, this has yet to be studied. Here, we summarize the recent advances in understanding the host-pathogen interaction of A. baumannii and propose a potential role of the IL-17 pathway in generating a protective immune response.

Change of plasma microRNA-208 level in acute myocardial infarction patients and its clinical significance.

BACKGROUND: To study the change of cardiac-specific microRNA-208 (miRNA-208) level in acute myocardial infarction (AMI) patients and to explore the role of miRNA-208 in AMI progression. METHODS: The consecutive subjects including 42 AMI patients, 22 patients with unstable angina (UA), and 40 healthy subjects in our hospital from January 2013 to December 2013 were enrolled in this study. The peripheral miRNA-208 level was measured with real-time reverse-transcription polymerase chain reaction (RT-PCR), and the plasma cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB) levels were determined using electrogenerated chemiluminescence (ECL) method. Patients in the AMI group were further grouped according to the number of stenosed coronary vessels and primary percutaneous coronary intervention (PCI) or not, and the difference in peripheral miRNA-208 level among these subgroups was analyzed. RESULTS: The miRNA-208 level was significantly higher in AMI group than in UA group and healthy controls immediately after admission (P<0.01). In the AMI patients, the plasma miRNA-208 level had a positive correlation with serum cTnI level (r=0.700, P=0.000). In 24 AMI patients who had undergone coronary angiography, the expression of miRNA-208 was significantly higher in patients with two or three stenosed coronary vessels. In 17 AMI patients who had successfully received emergent PCI treatment, the 24-h plasma miRNA-208 level was significantly lower than that immediately after admission (P<0.01). CONCLUSIONS: The peripheral plasma miRNA-208 level remarkably increases after cardiac infarction and may dramatically change along with the increase of the myocardial damage. Thus, it may be a new biomarker for AMI.

Genetic variants of the class A scavenger receptor gene are associated with essential hypertension in Chinese.

BACKGROUND: The class A scavenger receptor, which is encoded by the macrophage scavenger receptor 1 (MSR1) gene, is a pattern recognition receptor (PPR) primarily expressed in macrophages. It has been reported that genetic polymorphisms of MSR1 are significantly associated with many cardiovascular events. However, whether it links genetically to essential hypertension (EH) in Chinese is not defined. METHODS: We performed an independent case-control study in a Chinese population consisting of 617 EH cases and 620 controls by genotyping three single nucleotide polymorphisms (SNPs) of MSR1. RESULTS: We found that rs13306541 and rs3747531 were significantly associated with an increased risk of EH with per allele odds ratio (OR) of 1.63 [95% confidence interval (CI): 1.27-2.09; P<0.001] and 1.29 (95% CI: 1.09-1.52; P=0.003), respectively. Individuals with 2-4 risk alleles had a 2.03-fold (95% CI: 1.48-2.78) increased risk of EH compared with those having none of the risk alleles (P for trend <0.001). CONCLUSIONS: Our results indicate that genetic variants of MSR1 may serve as predictive markers for the risk of EH in combination with traditional risk factors of EH in Chinese population.

Diagnostic Accuracy of Natriuretic Peptides for Heart Failure in Patients with Pleural Effusion: A Systematic Review and Updated Meta-Analysis.

BACKGROUND: Previous studies have reported that natriuretic peptides in the blood and pleural fluid (PF) are effective diagnostic markers for heart failure (HF). These natriuretic peptides include N-terminal pro-brain natriuretic peptide (NT-proBNP), brain natriuretic peptide (BNP), and midregion pro-atrial natriuretic peptide (MR-proANP). This systematic review and meta-analysis evaluates the diagnostic accuracy of blood and PF natriuretic peptides for HF in patients with pleural effusion. METHODS: PubMed and EMBASE databases were searched to identify articles published in English that investigated the diagnostic accuracy of BNP, NT-proBNP, and MR-proANP for HF. The last search was performed on 9 October 2014. The quality of the eligible studies was assessed using the revised Quality Assessment of Diagnostic Accuracy Studies tool. The diagnostic performance characteristics (sensitivity, specificity, and other measures of accuracy) were pooled and examined using a bivariate model. RESULTS: In total, 14 studies were included in the meta-analysis, including 12 studies reporting the diagnostic accuracy of PF NT-proBNP and 4 studies evaluating blood NT-proBNP. The summary estimates of PF NT-proBNP for HF had a diagnostic sensitivity of 0.94 (95% confidence interval [CI]: 0.90-0.96), specificity of 0.91 (95% CI: 0.86-0.95), positive likelihood ratio of 10.9 (95% CI: 6.4-18.6), negative likelihood ratio of 0.07 (95% CI: 0.04-0.12), and diagnostic odds ratio of 157 (95% CI: 57-430). The overall sensitivity of blood NT-proBNP for diagnosis of HF was 0.92 (95% CI: 0.86-0.95), with a specificity of 0.88 (95% CI: 0.77-0.94), positive likelihood ratio of 7.8 (95% CI: 3.7-16.3), negative likelihood ratio of 0.10 (95% CI: 0.06-0.16), and diagnostic odds ratio of 81 (95% CI: 27-241). The diagnostic accuracy of PF MR-proANP and blood and PF BNP was not analyzed due to the small number of related studies. CONCLUSIONS: BNP, NT-proBNP, and MR-proANP, either in blood or PF, are effective tools for diagnosis of HF. Additional studies are needed to rigorously evaluate the diagnostic accuracy of PF and blood MR-proANP and BNP for the diagnosis of HF.

A preliminary study on the relationship between circulating tumor cells count and clinical features in patients with non-small cell lung cancer.

BACKGROUND: To evaluate the clinical value of circulating tumor cells (CTC) count in peripheral venous blood of patients with non-small cell lung carcinoma (NSCLC). METHODS: A total of 50 NSCLC patients who were diagnosed in Wuxi No. 2 People's Hospital from January 2013 to December 2013 were selected as the NSCLC group, 35 patients with lung benign tumor as the benign group, and 28 healthy subjects as the normal control group. Venous blood samples (3 mL) were collected in all subjects for counting the CTC, and a result of ≥8.7 was judged to be positive. The relationships between the positive rate of CTC and the age, sex, pathological type, and clinical stage of NSCLC were analyzed. RESULTS: CTC count was significantly higher in NSCLC group than in benign group and normal control group. In NSCLC patients, CTC count was not significantly correlated with sex, age, or the pathological type (P>0.05) but was closely related to clinical stage (P<0.01). Among NSCLC patients, CTC count significantly increased along with tumor progression. CONCLUSIONS: CTC count shows certain correlation with the clinical features of NSCLC and thus can, to certain extent, reflect the status of the disease.

Comparison of four DNA extraction methods for detecting Mycobacterium tuberculosis by real-time PCR and its clinical application in pulmonary tuberculosis.

OBJECTIVES: China is one of the countries with a high burden of Mycobacterium tuberculosis (MTB) infection. One challenge for the earlier diagnosis of tuberculosis is the DNA extraction of MTB. This study was to compare four MTB DNA extraction methods, and use the best one in the diagnosis of pulmonary tuberculosis. METHODS: A total of 43 serum and 94 plasma samples were collected from 124 clinical diagnosed pulmonary tuberculosis patients. Four different MTB DNA extraction methods, including phenol-chloroform method, Qiagen kit, Omega kit and magnetic bead method, were compared to determine which method displayed the highest sensitivity. A quantitative fluorescent PCR assay was also designed for the detection of MTB DNA. RESULTS: The highest DNA extraction efficiency (52.8%) and the best reproducibility (coefficient of variance =26.7%) were observed using the magnetic bead method. For 39 of the 124 (31.5%) pulmonary tuberculosis patients, MTB DNA was detected in their plasma or serum samples. Interestingly, 35.3% (12/34) of smear-negative cases were MTB DNA positive. CONCLUSIONS: In conclusion, magnetic bead method is the best one for the DNA extraction of MTB. The detection of MTB DNA may provide valuable information for the diagnosis of acid-fast bacilli (AFB) negative pulmonary tuberculosis patients.

Rational design of a thrombin electrochemical aptasensor by conjugating two DNA aptamers with G-quadruplex halves.

A novel strategy for the fabrication of an electrochemical aptasensor is proposed; this strategy has been employed in this work to assay thrombin concentration. Two well-designed oligonucleotides were used as the core element. G-quadruplex-hemin complexes can be formed on the surface of the electrode to give a detectable signal only when thrombin is not bound to the aptamers. The detection limit of the biosensor has been lowered to 10nM. Moreover, since the electroactive probe is not required to be bound to the oligonucleotide, this strategy may integrate the advantages of being both label-free and cost-effective.

Prediction and characterization of the linear IgE epitopes for the major soybean allergen ß-conglycinin using immunoinformatics tools.

The α subunit of ß-conglycinin is a major allergen in soybean. The objective of this study was to predict and identify the linear immunoglobulin (Ig)E epitopes of the soybean α subunit of ß-conglycinin. Three immunoinformatics tools were used to predict the potential epitopes and were confirmed by dot-blot inhibition using sera from soybean allergic subjects. As a result, 15 peptides were predicted and assembled by solid-phase synthesis. Eleven epitopes were identified by the dot-blot inhibition test. Moreover, peptide 3 had IgE binding capability with all sera(5/5) tested, while peptide 1, 4, 6, 8 and12 could bind to 4/5 of the sera samples. Secondary structure prediction of peptide 3 and circular dichroism test validated that the structure of peptide 3 was a random coil.

A new strategy based on aptasensor to time-resolved fluorescence assay for adenosine deaminase activity.

Quantitative protein bioanalysis in complex biological fluids presents considerable challenges in biological studies and medical diagnosis. The major obstacles are the background signals from the biological fluids and sensors themselves. Because the europium ion (Eu (III)) has the much longer fluorescence lifetime (1 ms) than that of the background (5 ns), time-resolved method can be widely used to eliminate the biological background. So, we report here an aptamer-based sensor (aptasensor) for time-resolved fluorescence assay of adenosine deaminase (ADA). This aptasensor employs two oligonucleotides labeled with DIG and biotin, respectively. The DNA1 (an oligonucleotide modified with biotin) is immobilized at a streptavidin-modified plate surface via the biotin-avidin bridge, and the DIG which is modified on the DNA2 serves as an affinity tag for the Eu(3+) labeled anti-DIG (Eu-anti-DIG) binding. If the adenosine is binding with DNA1, it will make the DNA1 in the closed state with a close-packed tight structure, which forbids the DNA2 approaching. And if the ADA is added into the mixture, the DNA1 unbends, because of the adenosine is transformed to inosine catalyzed by the ADA. Then DNA2 could hybridize with DNA1. Accordingly, the DIG finds Eu-anti-DIG and the Eu-anti-DIG will give a remarkable fluorescent signal. The detection limit of the aptasensor can be lowered to 2 UL(-1), which can meet the clinical requirement of ADA cutoff value (4 UL(-1)). Moreover, we were able to detect ADA in human serum quantitatively. Combined with time-resolved based measurements and aptasensor, this strategy holds great potential in protein analysis.

Performance assessment of the CapitalBio mycobacterium identification array system for identification of mycobacteria.

The CapitalBio Mycobacterium identification microarray system is a rapid system for the detection of Mycobacterium tuberculosis. The performance of this system was assessed with 24 reference strains, 486 Mycobacterium tuberculosis clinical isolates, and 40 clinical samples and then compared to the "gold standard" of DNA sequencing. The CapitalBio Mycobacterium identification microarray system showed highly concordant identification results of 100% and 98.4% for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM), respectively. The sensitivity and specificity of the CapitalBio Mycobacterium identification array for identification of Mycobacterium tuberculosis isolates were 99.6% and 100%, respectively, for direct detection and identification of clinical samples, and the overall sensitivity was 52.5%. It was 100% for sputum, 16.7% for pleural fluid, and 10% for bronchoalveolar lavage fluid, respectively. The total assay was completed in 6 h, including DNA extraction, PCR, and hybridization. The results of this study confirm the utility of this system for the rapid identification of mycobacteria and suggest that the CapitalBio Mycobacterium identification array is a molecular diagnostic technique with high sensitivity and specificity that has the capacity to quickly identify most mycobacteria.

Increased Siglec-1 expression in monocytes of patients with primary biliary cirrhosis.

OBJECTIVES: To evaluate Siglec-1 protein (CD169) and mRNA levels in peripheral blood monocytes of patients with primary biliary cirrhosis (PBC) and investigate its role in PBC pathogenesis by looking for correlations between Siglec-1 expression and key PBC associated biochemical indices. METHODS: FACS analysis was used to identify the percentage of peripheral blood monocytes positive for both CD14 and Siglec-1 in (a) 45 PBC patients, (b) 40 patients with liver cirrhosis after hepatitis B infection and (c) 36 healthy controls. Siglec-1 mRNA was measured by real-time RT-PCR and serum biomarkers by routine biochemistry. RESULTS: The percentage of CD14-Siglec-1 double positive cells was significantly higher (p< 0.01) in PBC patients than in healthy controls or cirrhosis post-hepatitis patients (13.68 +/- 2.44%, 1.0 +/- 0.2 %, and 4.1 +/- 0.5 %, respectively). Siglec-1 mRNA expression in the PBC group was 3.42 times higher than in healthy controls (p < 0.01). CONCLUSION: We investigated the role of Siglec-1 in PBC by assessing its expression in mononuclear cells of PBC patients and levels of secreted cytokines in cell supernatants after Siglec-1 RNA interference. It is possible that elevated Siglec-1 expression in peripheral blood monocytes of PBC patients is correlated with monocyte-mediated inflammatory responses during the development of PBC.

[Prevalence and drug tolerance of mycoplasma in patients with urogenital inflammation].

OBJECTIVE: To observe the prevalence and drug tolerance of mycoplasma(Ureaplasma urealyticum and Mycoplasma hominis) in patients with urogenital inflammation. METHODS: Three thousand and fifty-five patients with urogenital inflammation such as non-gonococcal urethritis(NGU), chronic prostatitis or pelvic inflammation from 1999 to 2003 were included. The results of mycoplasma culture and drug sensitivity test were analyzed. RESULTS: A total of 992(32.5%) cases were mycoplasma positive in the 3,055 patients, and there was no significant difference in the yearly positive percentage in the 5 years (P < 0.05). Among them, 701(70.7%) were infected with Ureaplasma urealyticum, 44(4.4%) with Mycoplasma hominis, and 247(24.9%) with both Ureaplasma urealyticum and Mycoplasma hominis, the Ureaplasma urealyticum infection rate being much higher than that of Mycoplasma hominis and mixed infection (P < 0.01). The high colony counting(> or = 10(4) cfu/ml) in Ureaplasma urealyticum infection patients accounted for 76.7%, while Mycoplasma hominis infection represented only 18.2%. The results of drug tolerance test showed a higher sensitivity to doxycycline, pristinamycin, josamycin and tetracycline (94.3%, 96.6%, 86.5% and 97.4% respectively), and a lower sensitivity to erythromycin and ofloxacin (54.8% and 29.4% respectively). CONCLUSIONS: Ureaplasma urealyticum and Mycoplasma hominis should be detected simultaneously and the drug tolerance test is needed for the selection of appropriate antibiotics.