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Extracellular vesicles (EVs) are actively secreted by mammalian cells. They are increasingly recognized as promising circulating biomarkers of disease progression. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is currently one of the most powerful techniques for the rapid analysis of biological samples, especially for discovering biomarkers for disease diagnosis and prognosis. It is unclear what cell culture medium components and EV isolation methods are suitable for MALDI-TOF MS analysis. Using a human lung carcinoma cell line (A549), we investigated and optimized the critical experimental conditions for EVs' protein profiling by combining differential ultracentrifugation and MALDI-TOF MS. The results demonstrated that medium components and ultracentrifugation procedures to extract EVs played important roles in MS detection. Compared with EV-depleted serum and normal serum medium, conditioned medium with 2% fetal bovine serum in this study maintained cell proliferation and displayed significant protein profiling of EVs. RPS27A (ribosomal protein), which plays an essential role in mRNA translation and ribosome assembly for the differentiation of cancer cells, was detected from the EVs of lung cancer cells associated with cancer cell migration and invasion. We also found the known tumor diagnosis marker, which is S100A10_S100 calcium-binding protein A10. Therefore, MALDI-TOF MS-based EV analysis with optimized experimental protocols can contribute to future development of rapid screening techniques of protein biomarkers associated with early cancer diagnosis.

Methods Forty-four COVID-19 patients (severe/critical: N = 8, non-severe: N = 36) were examined by next generation sequencing (NGS) of nasopharyngeal test paper to observe the effect of novel coronavirus infection to the microbial composition in upper airway. Results In these nasopharyngeal test paper samples, 38 kinds of bacteria, 10 kinds of viruses except SARS-CoV-2, nine kinds of fungi and three kinds of atypical pathogens had been found. There was some difference in microbial composition in the upper airway between severe and non-severe cases. Summary These results are important for us to study the effect of SARS-CoV-2 on the local microbial composition of upper airway and prevent opportunistic infection in severe patients.

A new troodontid (LH PV39) recovered from the Upper Cretaceous Wulansuhai Formation, Inner Mongolia, China, is described, highlighting the dorsoventrally compressed sacral centra. The completely fused neurocentral junctions indicate that LH PV39 had reached adulthood at the time of death, but its size is nevertheless 20% smaller than that of the sympatric Philovenator, demonstrating that it is the second small-bodied troodontid recovered from the Wulansuhai Formation. Phylogenetic analyses scoring LH PV39 using different strategies and performed with different algorithms unambiguously recovered it as a troodontid. While the parsimony-based analysis scoring LH PV39 as an independent OTU with all of its available characteristics included recovered it as a basal troodontid, the Bayesian analysis suggests a closer relationship of LH PV39 to Almas and an unnamed troodontid from Ukhaa Tolgod, Mongolia (MPC-D100/1126+D100/3500). Body size analysis confirmed a single trend of gigantism throughout the evolution of troodontids, and suggests that the Late Cretaceous troodontids evolved in two directions: (i) several size-independent characteristics evolved while retaining the small sizes that are typical of the Early Cretaceous relatives, resulting in the Late Cretaceous small-bodied troodontids; and (ii) size-dependent characteristics (e.g., the elongation of the rostrum) evolved accompanying the size increase, resulting in large-bodied derived troodontids. The mosaic features of the Late Cretaceous small-bodied troodontids place them intermediate between their Early Cretaceous basal relatives and the Late Cretaceous large-bodied taxa in a well-resolved phylogeny, which is crucial for understanding the size and morphological evolution of troodontids.

Polycyclic aromatic hydrocarbons (PAH) are well-known to be carcinogenic and the mechanisms that it contributes to oxidative DNA damage and aryl hydrocarbon receptor (AhR)-dependent induction are also well understood. However, little is known about the associations between PAH exposure, AhR expression, and oxidative DNA damage. We investigated their associations of AhR expression and oxidative DNA damage related to PAH exposure among 310 workers from a coke-oven plant in China. Urine biomarkers of PAH exposure (2-hydroxynaphthalene, 2-NAP; 2-hydroxyfluorene, 2-FLU; 9-hydroxyphenanthren, 9-PHE; and 1-hydroxypyrene, 1-OHP) and a marker of oxidative damage (8-hydroxy- 2′- deoxyguanosine, 8-OHdG) were measured by high performance liquid chromatography. AhR expression in venous blood was measured by reverse transcription -polymerase chain reaction. The results showed that increasing levels of urinary 1-OHP was positively associated with high 8-OHdG (OR (95% CI) was 4.01 (1.41–11.45) for 4th quartile, compared with 1st quartile, P for trend = 0.013). The similar associations were also found between urinary 1-OHP and high-AhR expressions (4th vs. 1st quartile = 3.50, 95% CI: 1.24–9.87, P for trend = 0.029). A significant association between AhR expression and high 8-OHdG was also found (4th vs. 1st quartile = 2.44, 95% CI: 1.05–5.70, P for trend = 0.027). In addition, mediation analysis showed the AhR expression could explain 35.9% of the association of oxidative DNA damage related to PAH exposure. Our findings implicated that the association between PAH exposure and oxidative DNA damage may be mediated by AhR expression among Chinese occupational workers.

Myocardial infarction (MI), which is generally known as a heart attack, occurs when there is a lack of blood supply to the heart. Electrocardiogram (ECG) is commonly used for the initial screening of MI but is not sufficiently sensitive. With ECG, cardiac biomarkers help to efficiently assess MI. The cardiac troponin I level has a high correlation with MI and is one of the gold standard biomarkers to identify MI. This study focuses on detecting cardiac troponin I by its parent antibody on a microcomb interdigitated electrode (IDE) sensor. To improve the detection, silica nanoparticles (SiNps) were complexed with antibody and immobilized on the IDE sensing surface. Amine-modified SiNps help to hold more antibodies on the sensing surface and interact with more cardiac troponin I molecules, leading to sensitive detection with a limit of detection of 1 fM (24 pg/mL). Moreover, the control proteins were failed to specifically interact, indicating the genuine detection of cardiac troponin I. This study helps to more easily and efficiently quantify the cardiac troponin I level to diagnose MI.

Myocardial infarction (MI) is considered as one of the major life-threatening health issues worldwide. Growing number of cases every year is demanding rapid, portable, and early detection by the sensing devices for the identification of MI. This research work introduces a modified interdigitated electrode (IDE) sensing surface constructed with single-walled carbon nanotube (SWCN) to detect the cardiac biomarker, C-reactive protein (CRP). CRP-specific aptamer was conjugated with gold nanoparticle and attached on SWCN-constructed IDE surface. This probe-modified sensing surface has reached the limit of CRP detection to 10 pM on a linear regression curve with the regression coefficient of R² = 0.9223 [y = 0.9198x - 0.4326]. Further, control molecules, such as random aptamer sequence and nontarget cardiac biomarker (Troponin I), did not show the current response, indicating the specific CRP detection. This sensing strategy helps to detect the lower level of CRP and diagnose the MI at its earlier stages.

Rheumatoid arthritis (RA), a chronic inflammatory disease, deprives patients' walking ability and reduces their life quality worldwide. Though recent studies have indicated the role of long noncoding RNA (lncRNA) ZFAS1 in several diseases, however, its role in RA remains uncharacterized. The present study aimed to unravel the the effect of ZFAS1 on RA. Herein, the RA mouse model and the human RA synoviocyte MH7A cell lines stimulated with TNF-α were established. ZFAS1 was next determined to be highly expressed in the mice with RA-like symptoms and TNF-α-stimulated MH7A cells while inhibiting ZFAS1 was demonstrated to promote proliferation and suppress apoptosis of MH7A cells. Furthermore, ZFAS1 knockdown exerted anti-inflammation effect in vitro and in vivo and reduced the arthritis index value. Moreover, RNA immunoprecipitation and dual-luciferase reporter assays identified the binding of ZFAS1 to microRNA (miR)-296-5p as well as the binding of miR-296-5p to matrix metalloproteinase-15 (MMP-15). Of note, ZFAS1 could bind miR-296-5p to up-regulate the expression of MMP-15. Our results from in vitro and in vivo experiments demonstrated silencing ZFAS1 mitigated RA-like symptoms such as inflammation and hyperplasia via miR-296-5p-dependent inhibition of MMP-15. Taken altogether, our study confirmed that ZFAS1 involved in RA progression by competitively binding to miR-296-5p and regulating MMP-15 expression.

A new small-bodied troodontid (LH PV39) recovered from the Upper Cretaceous Wulansuhai Formation, Suhongtu, Inner Mongolia, China, is described. The new specimen preserves six postaxial cervical vertebrae, five completely fused sacral and four posterior caudal vertebrae in addition to two manual unguals. The completely fused neurocentral junctions indicate that a skeletally mature individual of the same species of LH PV39 would be smaller than Philovenator and comparable in body size to a skeletal mature individual of Almas. The extremely dorsoventrally compressed sacral centra and neural canal, and the middle three sacral centra that are shorter and wider than the first and the last one distinguishing LH PV39 from other known troodontids. A series of phylogenetic analyses were conducted using modified published matrices. By coding LH PV39 in different strategies, the troodontid affinity of LH PV39 is confirmed and it was recovered as the sister taxon of either Mei and Sinovenator (LH PV39 scored as a separate OTU) or Linhevenator (incorporating LH PV39 into Philovenator) in the best resolved coelurosaurian interrelationships. The referral of LH PV39 to Philovenator does not seriously alter the phylogenetic position of Philovenator nor the interrelationships of troodontids. This new finding confirms that the small and large sized troodontids are coexisted in the Gobi Desert of the Mongolia Plateau until the end of Cretaceous.

To provide an elaborate understanding on the performance of Entererococcus sp. L1 in biogas production at different fermentation periods under controlled low temperature, biogas optimization experiments was undertaken. Specifically, microscopic and culture techniques, physiological and biochemical studies, and measurement of optical densities at 600 wavelengths (OD600) were undertaken to identify and characterize the Entererococcus sp. L1. The aim was to identify the strain and determine its optimum working conditions in biogas production at different fermentation periods under controlled low temperature. The result confirmed the strain as hydrogen-producing bacteria (Enteroccus sp. L1). It was also confirmed that the organic nitrogen source was significantly better than the inorganic nitrogen source in supporting the growth of the bacteria. The optimal pH and temperature for the growth and biogas production performance of the Entererococcus sp. L1 was 7 and 15 °C respectively. The Na2HPO4 concentration of 0.2% was confirmed to be best for the optimum growth of the bacteria, and addition of citric acid resulted to significant inhibitory effect on the growth of the Entererococcus sp. L1. The effect of adding different concentrations (0, 1%, 5%, 10%) of the L1 bacteria inoculum on cow dung at simulated low temperature (15 °C) biogas fermentation showed that 10% treatment had the highest gas production of 3121 ml, which was 89% higher than the control (0 concentration). The daily gas production rate was 0.02 m3/(m−3d−1), which was 90% higher than the control. This showed that 10% treatment provided the best gas production result, with highest gas production occurring at 10% treatment within 31–60 days of fermentation (p

Our understanding of spider reproductive biology is hampered by the vast anatomical diversity and difficulties associated with its study. Although authors agree on the two general types of female spider genitalia, haplogyne (plesiomorphic) and entelegyne (apomorphic), our understanding of variation within each group mostly concerns the external genital part, while the internal connections with the reproductive duct are largely unknown. Conventionally and simplistically, the spermathecae of haplogynes have simple two-way ducts, and those of entelegynes have separate copulatory and fertilization ducts for sperm to be transferred in and out of spermathecae, respectively. Sperm is discharged from the spermathecae directly into the uterus externus (a distal extension of the oviduct), which, commonly thought as homologous in both groups, is the purported location of internal fertilization in spiders. However, the structural evolution from haplo- to entelegyny remains unresolved, and thus the precise fertilization site in entelegynes is ambiguous. We aim to clarify this anatomical problem through a widely comparative morphological study of internal female genital system in entelegynes. Our survey of 147 epigyna (121 examined species in 97 genera, 34 families) surprisingly finds no direct connection between the fertilization ducts and the uterus externus, which, based on the homology with basal-most spider lineages, is a dead-end caecum in entelegynes. Instead, fertilization ducts usually connect with a secondary uterus externus, a novel feature taking over the functional role of the plesiomorphic uterus externus. We hypothesize that the transition from haplo- to entelegyny entailed not only the emergence of the two separate duct systems (copulatory, fertilization), but also involved substantial morphological changes in the distal part of the oviduct. Thus, the common oviduct may have shifted its distal connection from the uterus externus to the secondary uterus externus, perhaps facilitating discharge of larger eggs. Our findings suggest that the conventional model of entelegyne reproduction needs redefinition.

Previous studies in our laboratory demonstrated that Ring2 may affect DNA damage and repair through pathways other than through regulating the expression of the nucleotide excision repair protein. In a series of experiments using wild-type cell (16HBE and WI38) and small interfering RNA (siRNA) Ring2 cells exposed to benzo[a]pyrene (BaP), we evaluated the cell cycle and DNA damage. The benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE–DNA) adduct assay demonstrated that in vitro exposure to BaP increased DNA damage in a time- and dose-dependent manner in wild-type and siRNA Ring2 cells. Analysis of covariance showed that a decrease of Ring2 caused DNA hypersensitivity to BaP. Flow cytometry results and proliferating cell nuclear antigen levels indicated that inhibition of Ring2 attenuated the effect of BaP on S-phase arrest. Taken together, these data implied that the lower proportion of cells in the S phase induced by inhibition of Ring2 may play an important role in DNA hypersensitivity to BaP.

The Enantiornithes were the most taxonomically diverse bird group in the Mesozoic. Most of the known taxa are from Lower Cretaceous deposits of the Jehol Group in north-eastern China. A new specimen from the Jiufotang Formation in Jianchang, Liaoning Province, is described here; being a subadult individual at the time of death it had reached a relatively large size. The presence of uncinate processes, bicapitate and forked vertebral ribs, sternal ribs that were all of similar length, as well as the location of parapophyses and diapophyses on the thoracic vertebrae, may imply a rigid and volume-constant lung, and less efficient lung ventilation in enantiornithines. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113, 820–827. ADDITIONAL KEYWORDS: Jianchang – Jiufotang Formation – Liaoning province – respiratory apparatus – ribs – thoracic vertebrae.

The vacuolar-type H+ adenosine triphosphatase (V-ATPase) plays an important role in cellular acidification and bone resorption by osteoclasts. However, the direct effect of bovine parathyroid hormone (bPTH) on V-ATPase has not yet been elucidated. The aim of the present study was to assess the effects of bPTH on V-ATPase and osteoclasts. Osteoclasts from bone marrow (BM)-derived monocytes of C57BL/6 mice were cultured with or without bPTH. The mRNA and protein expression levels of the V-ATPase a3-subunit and d2-subunit (by RT-qPCR and western blot analysis), V-ATPase activity (using the V type ATPase Activity Assay kit) and the bone resorption function of osteoclasts (by bone resorption assay) were examined following treatment with various concentrations of bPTH (0.1, 1.0, 10 and 100 ng/ml) alone or with bPTH and its inhibitor, bafilomycin A1. Furthermore, the expression of parathyroid hormone (PTH) receptors in osteoclasts was also detected. The results revealed that the mRNA and protein expression levels of V-ATPase a3-subunit and d2-subunit increased in a dose‑dependent manner, paralleling the level of bPTH present. In addition, an increase in the concentration of bPTH was accompanied by the increased resorption capability of osteoclasts, whereas bone resorption was inhibited in the presence of bafilomycin A1. In addition, we confirmed the existence of parathyroid hormone 1 receptor (PTH1R) in osteoclasts using three different methods (RT-qPCR, western blot analysis and immunofluorescence staining). We found that bPTH enhanced the bone resorption capability of osteoclasts by modulating the expression of V-ATPase subunits, intracellular acidification and V-ATPase activity. Thus, we propose that PTH has a direct effect on osteoblasts and osteoclasts, and that this effect is mediated through PTH1R, thus contributing to bone remodeling.

The aim of this study was to explore the expression and clinical significance of miR-494 and PTEN (phosphatase and tensin homologue deleted on chromosome ten) in non-small cell lung cancer (NSCLC). Immunohistochemistry for PTEN and in situ hybridization (ISH) for miR-494 were performed in 92 NSCLC tissues and 10 normal lung tissues to detect their expression, and correlation between their expression with clinical characteristics and prognosis was analyzed. The expression of miR-494 was significantly higher in NSCLC than in normal lung tissues (P = 0.004). The positive expression of PTEN protein in the lung carcinoma tissues was significantly lower than that in the normal lung tissues (P = 0.013), while the level of miR-494 expression was negatively correlated with PTEN expression (r = −0.577, P