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A shale reservoir’s gas content is an indispensable, crucial parameter used in exploration, formation evaluation, block selection and exploitation. Canister gas desorption measurements are used to quantify lost gas volumes that are key to determine the gas content of the reservoir Usually, a lost gas volume is evaluated using the United States Bureau of Mines (USBM) direct method and/or a polynomial curve-fitting method. These methods extrapolate desorption data to time zero in the curve of a cumulative desorption volume versus the square root of time. Lost gas time is the time spent lifting the core out of a hole, handling and placing it inside a canister for evaluation., For cores with densely developed natural fractures, neither the USBM method nor the polynomial curve-fitting method can accurately estimate the lost gas volume if the lifting, handling and canister storage takes a long time. The lost gas period is a non-isothermal gas desorption process; the surrounding temperature keeps declining. Experiments were conducted to determine the methane desorption volumes of a Duvernay shale sample under various pressures and temperatures. Using Boyle’s law and the multilayer adsorption model, a novel mathematical model was built to calculate lost gas volume, including free gas and desorption gas. Because gas desorption is an endothermic procedure, equations for the inhaled energy were developed to overcome the van der Waals force between the molecules, deorbit the vibration equilibrium and the migration of molecules into the air. Finally, a shale sample with dense natural fractures from the Duvernay formation of Alberta, Canada, was used to validate the mathematical model. For comparison, the results from the USBM method, polynomial curve-fitting method and model’s calculations are illustrated in tables and figures. The outcomes exposed the results from the USBM method and polynomial curve-fitting method as deviating from the real gas content in the Duvernay formation without the consideration of the free gas loss. The model is reliable for performing gas content evaluations because it considers dual-pore structure, PVT behaviour and free gas escape simultaneously.

We experimentally study a circuit quantum acoustodynamics system, which consists of a superconducting artificial atom, coupled to both a two-dimensional surface acoustic wave resonator and a one-dimensional microwave transmission line. The strong coupling between the artificial atom and the acoustic wave resonator is confirmed by the observation of the vacuum Rabi splitting at the base temperature of dilution refrigerator. We show that the propagation of microwave photons in the microwave transmission line can be controlled by a few phonons in the acoustic wave resonator. Furthermore, we demonstrate the temperature effect on the measurements of the Rabi splitting and temperature induced transitions from high excited dressed states. We find that the spectrum structure of two-peak for the Rabi splitting becomes into those of several peaks, and gradually disappears with the increase of the environmental temperature $T$. The quantum-to-classical transition is observed around the crossover temperature $T_{c}$, which is determined via the thermal fluctuation energy $k_{B}T$ and the characteristic energy level spacing of the coupled system. Experimental results agree well with the theoretical simulations via the master equation of the coupled system at different effective temperatures., Comment: 14 pages, 6 figures

The mechanical conditions that allow seismogenic faults to be activated in distinctive positions and moments remain uncertain. An integrated approach is proposed to quantify the hydrological connection between stimulated wells and associated seismogenic faults during fracturing stimulation. A case study in Fox Creek, Alberta, was conducted to demonstrate the applicability of this integrated approach. Ant tracking was used to identify the subvertical basement-rooted faults from a three-dimensional seismic reflection survey. A coupled flow–geomechanics simulation was conducted to quantify the well–fault hydraulic connection in terms of pore pressure diffusion and poroelastic stress perturbation during fracturing stimulation. Three identified basement-rooted faults were corroborated by the focal mechanisms and spatial distribution of induced siemsicity events. The negative shear stress gradient indicates the downward shear growth during hydraulic connections. The induced earthquakes were triggered by fluid diffusion through hydraulic fractures along high-permeability fault damage zones downwards into the basement. This basal fault slip was attributed primarily to the elevated pore pressure along the fault plane in response to fracturing fluid injection. The moderate distance (879 m for NS-oriented wells and 749 m for NW-SE-oriented wells) between the future horizontal wellbores and critically-stressed faults could mitigate the effects of well-fault hydraulic connection and reduce the seismicity risks.

Hundreds of horizontal wells have been performed fracturing operations to exploit the unconventional shale gas resources in the Duvernay Formation of Fox Creek, Alberta. Despite achieving the practical analysis of shale gas production via the data-mining approach, previous studies failed to incorporate comprehensive site-specific geological and operational factors. In this study, a comprehensive machine-learning approach is developed to forecast the shale gas production via the integration of geological and operational factors. Thirteen geological and operational parameters deriving from the well logging, core experiment and treatment data are included as the input variables, whereas the 12-month shale gas production is regarded as the target variable. Results show that factors that mostly contributed to the shale gas production are found to be total fluid injection, total proppant mass, well TVD, permeability, y coordinate, porosity, gas saturation, number of stages, x coordinate, formation pressure, horizontal length, distance to fault and Duvernay thickness. Four machine learning methods are evaluated, where the Extra Trees approach has led to the highest coefficient of determination R2 of 0.81. Case study for Well 2 have shown that the shale gas production can be doubled if increase the total pumped volume and proppant placed mass by approximately 73% and 38%.

Helium-3, a potential fusion fuel which is widely deposited in the lunar regolith, may act as a leading role in the future energy supply. Despite abundant investigations of the lunar regolith properties, the research of the transport behavior of helium-3 in porous media is still lacking, which is essential for the prediction of its distribution characteristics. In this work, the storage mechanisms of helium-3 in the lunar regolith are firstly reviewed, and it is found that the primary driving force for the diffusion of helium-3 in porous media arises from its local supersaturation caused by the solar wind flux. After an estimation of the Knudsen number (Kn), it is concluded that Knudsen diffusion is the only transfer regime for the transport of helium-3 in the lunar subsurface because of a rarefaction effect. Moreover, the diffusion coefficient of helium-3 in porous media is predicted and the factors affecting this coefficient are analyzed. Furthermore, a sensitivity analysis is performed due to the uncertainty of the geometric factors of the lunar regolith. Finally, the storage and transport behavior for shale gas and helium-3 are compared, and the prospect for the mining of helium-3 is discussed.

Fracture interference has been frequently observed between the parent wells (i.e., the pre-existing wells) and the child wells (i.e., infill wells) in the unconventional tight/shale reservoirs with massive hydraulic fractures. Such interference usually decreases the expected productivity and capital efficiency of the child wells due to the overlapped drainage areas of the parent and child wells. In this study, a new procedure using machine learning (ML) methods is proposed to maximize the productivity of the child wells in Montney formation. More specifically, three types of parent-child well patterns are first defined based on production and trajectory and a comprehensive dataset is developed to include location information on the wells, fracturing parameters, and production history. Then, various strategies are employed to eliminate the redundant and nonsignificant variables and four ML methods including neural networks, random forests, gradient boosting decision trees and linear regression are introduced to predict the first-year production of child wells. Performances of the four ML methods are evaluated with anti-overfitting strategy and the random forests (RF) is found to be the most suitable one for this issue. Finally, a fine-tuned random forest model is applied to optimize the usage of proppant, the volume of fluid injected to get higher productivity of parent-child mode. In addition, the correlation among well spacing and first-year barrels of oil equivalent (BOE) of child well is also analyzed. The calculation results indicated that fracture interference decreases as the spacing increases and the recommend one is above 200 m. The results are also consistent with some actual observation data reported, which further demonstrates the feasibility and credibility of the method adopted.

Lung injury is one of the pathological hallmarks of most respiratory tract diseases including asthma, acute respiratory distress syndrome (ARDS) and chronic obstructive pulmonary disease (COPD). It involves progressive pulmonary tissue damages which are usually irreversible and incurable. Therefore, strategies to facilitate drug development against lung injury are needed. Here, we characterized the zebrafish folate-deficiency (FD) transgenic line that lacks a fully-developed swim bladder. Whole-mount in-situ hybridization revealed comparable distribution patterns of swim bladder tissue markers between wild-type and FD larvae, suggesting a proper development of swim bladder in early embryonic stages. Unexpectedly, neutrophils infiltration was not observed in the defective swim bladder. Microarray analysis revealed a significant increase and decrease of the transcripts for cathepsin L and a cystatin B (CSTB)-like (zCSTB-like) proteins, respectively, in FD larvae. The distribution of cathepsin L and the zCSTB-like transcripts was spatio-temporally specific in developing wild-type embryos and, in appropriate measure, correlated with their potential roles in maintaining swim bladder integrity. Supplementing with 5-formyltetrahydrofolate successfully prevented the swim bladder anomaly and the imbalanced expression of cathepsin L and the zCSTB-like protein induced by folate deficiency. Injecting the purified recombinant zebrafish zCSTB-like protein alleviated FD-induced swim bladder anomaly. We concluded that the imbalanced expression of cathepsin L and the zCSTB-like protein contributed to the swim bladder malformation induced by FD and suggested the potential application of this transgenic line to model the lung injury and ECM remodeling associated with protease/protease inhibitor imbalance.

Background Thrombomodulin (TM), an integral membrane protein, has long been known for its anticoagulant activity. Recent studies showed that TM displays multifaceted activities, including the involvement in cell adhesion and collective cell migration in vitro. However, whether TM contributes similarly to these biological processes in vivo remains elusive. Methods We adapted zebrafish, a prominent animal model for studying molecular/cellular activity, embryonic development, diseases mechanism and drug discovery, to examine how TM functions in modulating cell migration during germ layer formation, a normal and crucial physiological process involving massive cell movement in the very early stages of life. In addition, an in vivo assay was developed to examine the anti-hemostatic activity of TM in zebrafish larva. Results We found that zebrafish TM-b, a zebrafish TM-like protein, was expressed mainly in vasculatures and displayed anti-hemostatic activity. Knocking-down TM-b led to malformation of multiple organs, including vessels, heart, blood cells and neural tissues. Delayed epiboly and incoherent movement of yolk syncytial layer were also observed in early TM-b morphants. Whole mount immunostaining revealed the co-localization of TM-b with both actin and microtubules in epibolic blastomeres. Single-cell tracking revealed impeded migration of blastomeres during epiboly in TM-b-deficient embryos. Conclusion Our results showed that TM-b is crucial to the collective migration of blastomeres during germ layer formation. The structural and functional compatibility and conservation between zebrafish TM-b and mammalian TM support the properness of using zebrafish as an in vivo platform for studying the biological significance and medical use of TM. Electronic supplementary material The online version of this article (10.1186/s12929-019-0549-2) contains supplementary material, which is available to authorized users.

Figure S1. The alignment and identity of zTHBD primary sequence with those of zebrafish TM-a and TM-b. Figure S2. Efficiency of zTM-b MO. Figure S3. Schematic diagram of the predictive ETS-1 binding sites in TM promoter. Figure S4. The zTM-b morphants did not recover from the developmental anomalies as the embryogenesis proceeding to the later stages. Figure S5. In vitro activity assay for thrombomodulin. (PDF 1830 kb)

Gas adsorption can induce kerogen swelling with changes in pore volumes. In this study, hybrid Grand Canonical Monte Carlo/Molecular Dynamics simulations are conducted to investigate gas adsorption in kerogen and the corresponding sorption-induced swelling. A unified relationship between volumetric strain and an absolute adsorption amount is developed for different adsorbates. A theoretical model for calculating excess adsorption isotherms coupling swelling is proposed. Results show that (i) steep increases in C2H6 and CO2 adsorption isotherms at lower pressure indicate stronger affinities of C2H6 and CO2 than CH4; however, a larger size of C2H6 results in smaller accessible pore volumes and smaller maximum absolute adsorption amounts than CH4 and CO2. (ii) A linear relationship between volumetric strain and an adsorption amount is shown for CH4, C2H6 and CO2, separately. The volumetric strain caused by per unit of the absolute adsorption amount for C2H6 is the greatest because C2H6 with greater diameters has larger contact areas with pore walls for the same adsorbed amount. The maximum swelling upon CO2 adsorption is the largest (CO2 > CH4 > C2H6) due to its greatest absolute adsorption amount. (iii) Excess adsorption isotherms generated by our model are consistent with excess adsorption data calculated by variable pore volumes, which shows a large discrepancy compared to that determined using constant volumes, especially at high pressures. The difference between the fraction of free CH4 in our model and that calculated by using a constant volume is up to 23.2% at 323 K, 40 MPa. Therefore, this theoretical model can accurately determine free and adsorbed gas amounts in shale, further influencing prediction of hydrocarbon production and CO2 sequestration.

The dynamic imbibition behavior of fluids through nanopores has tremendous implications in the chemistry and engineering fields. The dynamics of water confined in nanopores significantly deviates from the macroscopic hydrodynamics in many aspects. In this work, we investigate the capillary dynamics of confined water in nanopores under a quasi-continuum flow regime, considering precursor films ahead of a main meniscus. A model for the imbibition behavior in nanopores is proposed, which takes the propagation of the precursor films into account. In addition, the increased viscosity of water near a solid wall and dynamic contact angle are incorporated in the proposed model. Experimental results are used to validate the model. It is found that the increased viscosity for water near a solid surface significantly reduces the fluidity, and the fluid loss from the surface water into the precursor films plays a key role in the imbibition behavior. Ignoring this effect will highly overestimate the true imbibition length. For a nano-channel with a width of 5 nm, the predicted imbibition length without the consideration of the fluid loss will almost be double of the true imbibition length. Moreover, this deviation becomes smaller with the increase of the pore size. Furthermore, the dynamic contact angle has a minimal effect on the imbibition length, which indicates an improvement of nano-channel-coating efficiency by the propagation of the precursor films. The proposed model captures important physics for capillary dynamics at the nanoscale, and has wide applications in many fields.

Understanding water imbibition behaviors in shale formations plays an essential role in shale gas development. The effective viscosity of water in shale nanopores is usually different from that of the bulk phase because of confined conditions. In this work, a model considering interactions between water and a solid surface is proposed to predict the effective viscosity of water at the nanoscale, which is inserted into the classic Lucas-Washburn (L-W) model to describe the water imbibition behavior in shale formations. This model is derived based on a molecular kinetic theory and incorporates the disjoining pressure. Published experimental data is used to validate the model. It is demonstrated that the effective viscosity of water in hydrophilic nanopores is much higher than that of the bulk phase because of strong interactions between water and a solid surface, and the deviation significantly increases when the separation is below 10 nm. For a hydrophilic capillary tube with a diameter of 10 nm, the water effective viscosity is approximately 2.5 times higher than that of the bulk phase. Moreover, this deviation is larger for a capillary tube compared with a capillary channel, because of the curvature effect. Besides, the effective viscosity for water under a hydrophobic condition is smaller than that of the bulk phase water, because the structural repulsive force dominates under a hydrophobic condition. This work establishes a theoretical foundation to calculate the effective viscosity for water flow at the nanoscale. Furthermore, it helps to understand fracturing fluid imbibition behavior in shale gas reservoirs, which will benefit the simulation of fluid flow at the reservoir scale.

There has been a remarkable increase in induced seismicity in Western Canada and the majority of these events can be related to hydraulic fracturing operations applied to develop unconventional reservoirs. The underlying mechanism of such induced seismicity is still unclear and needs to be investigated to mitigate risks of future events. In this paper, we propose a new integrated approach to characterize hydraulic fracturing-induced seismicity. Inferred faults in the target area were first identified by the ant-tracking approach and hydraulic fractures were characterized by simulating hydraulic fracturing processes. The coupled flow-geomechanics modeling was then performed to compute the Coulomb Failure Stress and determine the reactivation of pre-existing faults. A case study was finally utilized to demonstrate the applicability of the integrated method. Four north-south-oriented faults were interpreted and half-length of hydraulic fractures were calculated from 84 to 127 m. The calculated results of the flow-geomechanics model were in good agreement with the actual induced seismicity spatially and temporally. Three types of triggering mechanisms were accounted for HF-induced seismicity, including hydraulic fractures propagation, the connection between hydraulic fractures and a fault and connections of hydraulic fractures with the natural fractures around the fault. The injection rate could be decreased to mitigate risks of future seismic events.

The rock wettabilities and water contact angles describing interactions between CO2, CH4, brine and shale formations are of great significance to CO2 sequestration and enhanced gas recovery processes. However, water contact angles on the surfaces of shale organic matter in the atmospheres of CO2 and CH4 under reservoir conditions are not well-understood. In this study, we present an investigation of water/brine contact angles as functions of temperature, pressure, salinity, ion types, and gas contents by molecular dynamics simulations, and compare results with data from literature. It is found that temperature has profound effects on water contact angles below the critical temperature at an intermediate pressure. Meanwhile, water contact angles increase with pressure before reaching 180° at high pressure and the CO2-water-shale organic matter system turns from a neutrally-wet state to a CO2-wet state at the critical pressure of CO2. We also demonstrate that salinity and ion types have minor impacts on the brine contact angles in the CO2-brine-shale system. Only a slight increase in water contact angles is observed with increasing salinity, and an increase in brine contact angles caused by the divalent cations Mg2+ and Ca2+ is larger than that by the monovalent cations Na+ at the same salinity. Additionally, an increase in the CO2 fraction of gas mixtures can increase water contact angles at the same pressure and temperature. The surfaces of shale organic matter have a stronger affinity for CO2 than CH4, which contributes to a higher CO2 adsorption capacity and improves the displacement efficiency of CH4.

Background: Because of limited stability and sensitivity, circulating miRNAs as noninvasive biomarkers have not so far been used for early diagnosis and prognosis of non–small cell lung cancer (NSCLC) in clinic. Therefore, it is imperative to find more reliable biomarker(s). Methods: We performed one of most sensitive qRT-PCR assays, S-Poly(T) Plus, to select differently expressed miRNAs from genome-wide miRNA profiling. miRNA candidates were validated through a three-phase selection and two validation processes with 437 NSCLC cases and 415 controls. Results: A unique set of 7 and 9 miRNAs differed significantly in adenocarcinoma (ADC) and squamous cell carcinoma (SCC) samples compared with those in controls, of which, there were 5 universal biomarkers for NSCLC (ADC or SCC). Ten of 11 miRNAs could discriminate early stage (stage I) of NSCLC from healthy individuals. Risk score was obtained from the validation set-1 and was tested using the ROC curves with a high area under ROC curve of 0.89 in ADC and 0.96 in SCC. Ultimately, potential biomarkers and the risk score were verified by the validation set-2 with a sensitivity of 94% and a specificity of 91.6% in ADC, and a sensitivity of 98.5% and a specificity of 51.5% in SCC, respectively. Conclusions: Taken together, 7 miRNAs and 9 miRNAs may provide noninvasive biomarkers for diagnosis and prognosis in ADC and SCC, respectively. Impact: On the basis of our sensitive and accurate method, we hope that these candidate miRNAs may have strong impact on the early lung cancer diagnosis.

Background: The use of video-assisted thoracic surgery (VATS) in the treatment of thymoma is becoming more and more common. The aim of this study was to evaluate safety and effectiveness of thoracoscopic treatment of thymoma and, more importantly, to report long-term follow-up results of a large single-center cohort. Methods: A retrospective review of a prospective database was performed. In total, 150 patients who underwent VATS resection for thymoma at Peking University People’s Hospital from April 2001 to November 2014 were retrospectively reviewed. Results: Average operation time was 140.1±54.2 min, median blood loss was 50 mL (range, 10–700 mL), median post-operative drainage time was 3 days (range, 1–11 days), and median length of post-op stay was 5 days (range, 2–20 days); 134 patients (89.3%) were followed up successfully. Median follow-up was 59.5 months (range, 2–187 months). Five- and 10-year recurrence free survival (RFS) rates of entire group were 96.5% and 94.4%, respectively; 5- and 10-year RFS rates for Masaoka stages I + II were 98.1% and 98.1%, respectively; 5- and 6-year RFS rates for Masaoka stage III were 90% and 60%, respectively. One case of recurrence in ve Masaoka stage IV patients was observed, and 4-year RFS was 80%. Multivariable analysis indicated that recurrence tended to occur in Masaoka stages III + IV patients (P=0.037, HR =12.69, 95% CI: 1.17–138.22) and older patients had a lower risk of recurrence (P=0.029, HR =0.87, 95% CI: 0.77–0.99). Myasthenia gravis (MG) presented in 44 patients (29.3%), of which 36 patients (81.8%) were followed up. Nine patients achieved complete remission, and 19 patients had symptom improvement after surgery. Overall response rate of MG was 77.8% (28/36). Conclusions: VATS was a safe and effective procedure for treatment of thymomas with satisfactory prognosis. MG with thymoma treated by VATS had comparable neurological outcome to that associated with the trans-sternal approach.

Lung cancer is one of the most common cancers and the predominant cause of cancer‐related death in the world. The low accuracy of early detection techniques and high risk of relapse greatly contribute to poor prognosis. An accurate clinical tool that can assist in diagnosis and surveillance is urgently needed. Circulating tumor DNA (ctDNA) is free DNA shed from tumor cells and isolated from peripheral blood. The genomic profiles of ctDNA have been shown to closely match those of the corresponding tumors. With the development of approaches with high sensitivity and specificity, ctDNA plays a vital role in the management of lung cancer as a result of its reproducible, non‐invasive, and easy‐to‐obtain characteristics. However, most previous studies have focused on advanced lung cancer. Few studies have investigated ctDNA in the early stages of the disease. In this review, we focus on ctDNA obtained from patients in the early stage of lung cancer, provide a summary of the related literature to date, and describe the main approaches to ctDNA and the clinical applications.

Folate (vitamin B9) is an essential nutrient required for cell survival, proliferation, differentiation and therefore embryogenesis. Folate deficiency has been associated with many diseases, including congenital heart diseases and megaloblastic anemia, yet the mechanisms underlying these remains elusive. Here, we examine the impact of folate deficiency on the development of the circulation system using a zebrafish transgenic line which displays inducible folate deficiency. Impaired hematopoiesis includes decreased hemoglobin levels, decreased erythrocyte number, increased erythrocyte size and aberrant c-myb expression pattern were observed in folate deficient embryos. Cardiac defects, including smaller chamber size, aberrant cardiac function and cmlc2 expression pattern, were also apparent in folate deficient embryos. Characterization of intracellular folate content in folate deficiency revealed a differential fluctuation among the different folate derivatives that carry a single carbon group at different oxidation levels. Rescue attempts by folic acid and nucleotides resulted in differential responses among affected tissues, suggesting that different pathomechanisms are involved in folate deficiency-induced anomalies in a tissue-specific manner. The results of the current study provide an explanation for the inconsistent outcome observed clinically in patients suffering from folate deficiency and/or receiving folate supplementation. This study also supports the use of this model for further research on the defective cardiogenesis and hematopoiesis caused by folate deficiency.

Nanoparticles (NP) that target bone tissue were developed using PLGA-PEG (poly(lactic-co-glycolic acid)-polyethylene glycol) diblock copolymers and bone-targeting moieties based on aspartic acid, (Asp)(n(1,3)). These NP are expected to enable the transport of hydrophobic drugs. The molecular structures were examined by (1)H NMR or identified using mass spectrometry and Fourier transform infrared (FT-IR) spectra. The NP were prepared using the water miscible solvent displacement method, and their size characteristics were evaluated using transmission electron microscopy (TEM) and dynamic light scattering. The bone targeting potential of the NP was evaluated in vitro using hydroxyapatite affinity assays and in vivo using fluorescent imaging in zebrafish and rats. It was confirmed that the average particle size of the NP was

Folate is a nutrient essential for the development, function and regeneration of nervous systems. Folate deficiency has been linked to many neurological disorders including neural tube defects in fetus and Alzheimer's diseases in the elderly. However, the etiology underlying these folate deficiency-associated diseases is not completely understood. In this study, zebrafish transgenic lines with timing and duration-controllable folate deficiency were developed by ectopically overexpressing a recombinant EGFP-γ-glutamyl hydrolase (γGH). Impeded neural crest cell migration was observed in the transgenic embryos when folate deficiency was induced in early stages, leading to defective neural tube closure and hematopoiesis. Adding reduced folate or N-acetylcysteine reversed the phenotypic anomalies, supporting the causal link between the increased oxidative stress and the folate deficiency-induced abnormalities. When folate deficiency was induced in aged fish accumulation of beta-amyloid and phosphorylated Tau protein were found in the fish brain cryo-sections. Increased autophagy and accumulation of acidic autolysosome were apparent in folate deficient neuroblastoma cells, which were reversed by reduced folate or N-acetylcysteine supplementation. Decreased expression of cathepsin B, a lysosomal protease, was also observed in cells and tissue with folate deficiency. We concluded that folate deficiency-induced oxidative stress contributed to the folate deficiency-associated neuropathogenesis in both early and late stages of life.

A metal titanium coating was prepared in alumina foam by chemical vapor deposition method. The phase composition of the coating was analyzed by X-ray diffraction (XRD). The coverage of the coating on the alumina foam was analyzed by both optical and scanning electron microscope (SEM) with energy dispersive spectrometer (EDS). The results showed that the obtained titanium coating had a high purity, and showed a good coverage and uniform distribution on the skeleton surface of the foam. The skeleton surface became smooth due to coverage of the tiny holes and cracks by the coating.

Background Folate is an essential nutrient for cell survival and embryogenesis. 10-Formyltetrahydrofolate dehydrogenase (FDH) is the most abundant folate enzyme in folate-mediated one-carbon metabolism. 10-Formyltetrahydrofolate dehydrogenase converts 10-formyltetrahydrofolate to tetrahydrofolate and CO2, the only pathway responsible for formate oxidation in methanol intoxication. 10-Formyltetrahydrofolate dehydrogenase has been considered a potential chemotherapeutic target because it was down-regulated in cancer cells. However, the normal physiological significance of 10-Formyltetrahydrofolate dehydrogenase is not completely understood, hampering the development of therapeutic drug/regimen targeting 10-Formyltetrahydrofolate dehydrogenase. Methods 10-Formyltetrahydrofolate dehydrogenase expression in zebrafish embryos was knocked-down using morpholino oligonucleotides. The morphological and biochemical characteristics of fdh morphants were examined using specific dye staining and whole-mount in-situ hybridization. Embryonic folate contents were determined by HPLC. Results The expression of 10-formyltetrahydrofolate dehydrogenase was consistent in whole embryos during early embryogenesis and became tissue-specific in later stages. Knocking-down fdh impeded morphogenetic movement and caused incorrect cardiac positioning, defective hematopoiesis, notochordmalformation and ultimate death of morphants. Obstructed F-actin polymerization and delayed epiboly were observed in fdh morphants. These abnormalities were reversed either by adding tetrahydrofolate or antioxidant or by co-injecting the mRNA encoding 10-formyltetrahydrofolate dehydrogenase N-terminal domain, supporting the anti-oxidative activity of 10-formyltetrahydrofolate dehydrogenase and the in vivo function of tetrahydrofolate conservation for 10-formyltetrahydrofolate dehydrogenase N-terminal domain. Conclusions 10-Formyltetrahydrofolate dehydrogenase functioned in conserving the unstable tetrahydrofolate and contributing to the intracellular anti-oxidative capacity of embryos, which was crucial in promoting proper cell migration during embryogenesis. General significance These newly reported tetrahydrofolate conserving and anti-oxidative activities of 10-formyltetrahydrofolate dehydrogenase shall be important for unraveling 10-formyltetrahydrofolate dehydrogenase biological significance and the drug development targeting 10-formyltetrahydrofolate dehydrogenase.

Aims The nuclear hormone receptor estrogen-related receptor α (ERRα) regulates the activation of mitochondrial genes in various human tissues, but its role in the adrenal gland and its disorders has not been defined. Therefore, we examined ERRα expression in both normal adrenal cortex (NAC) and adrenocortical tumor (ACT) in order to study the possible correlation of ERRα with adrenal development and tumor development. Methods Human adrenal specimens (non-pathological fetal n = 7; non-pathological post-birth n = 40; aldosterone producing adenoma (APA) n = 11; cortisol producing adenoma (CPA) n = 11; adrenocortical carcinoma (ACC) n = 8) were immunohistochemically examined in this study. NAC (n = 13) and ACT (n = 28) frozen tissue specimens were also available for studying ERRα mRNA levels. Key findings In fetal NAC tissues, ERRα labeling index (LI) in fetal zone (FZ) was significantly higher that that in neocortex (NC), and the differences among age groups for overall mean LI was statistically significant when analyzed according to individual cortical layers. ERRα LI was also significantly higher in ACC than in other types of ACT. ERRα mRNA was detected in NAC and all types of ACT. Significance Results of our present study suggest a possible role of ERRα in adrenal development and ACC.

Capsulin and Musculin are basic helix-loop-helix transcription factors, but their biophysiological roles in zebrafish cranial myogenesis are unclear. Expressions of endogenous capsulin transcripts are detected at the central- (~24-hpf) and at dorsal- and ventral-mesoderm cores (~30-72 hpf) of branchial arches. In contrast, musculin transcripts are expressed as a two-phase manner: early phase (20-22 hpf) expressions of musculin are detected at the head mesoderm, whereas late-phase (36-72 hpf) are detected at all presumptive head-muscle precursors. Knockdown of either capsulin or musculin leads to loss of all cranial muscles without affecting trunk muscle development. The defective phenotypes of Capsulin- and Musculin-morphant can be rescued by co-injection of mRNA of each other. Both myf5 and myod transcripts are down-regulated in the Capsulin-morphant while myod transcripts are up-regulated in the Musculin-morphant. Therefore, we propose a putative regulatory network to understand how capsulin/musculin regulate distinctly either myf5 or myod during zebrafish craniofacial myogenesis.

Peroxisome proliferator-activated receptor-γ (PPARγ) is a nuclear receptor for the antidiabetic agent thiazolidinedione, which exerts various physiological activities, independent of lowering blood glucose. However, the role of PPARγ in aldosterone production has not been clarified. The objective of this study was to investigate the effect of PPARγ on aldosterone synthase gene (CYP11B2) expression and aldosterone production. Localization of PPARγ expression in normal adrenal cortex was determined by immunohistochemistry. Aldosterone production and CYP11B2 expression levels were determined using human adrenocortical carcinoma H295R cells. Pioglitazone suppressed angiotensin II-induced aldosterone secretion and CYP11B2 expression. PPARγ was expressed in zona glomerulosa in human normal adrenal gland. PPARγ overexpression enhanced pioglitazone-mediated CYP11B2 transrepression. The pioglitazone-mediated suppression of aldosterone secretion and CYP11B2 expression were canceled by PPARγ L466A/E469A mutant. Pioglitazone also suppressed potassium-mediated CYP11B2 induction, but not N6-2'-O-dibutyladenosine-3',5'-cyclic monophosphate stimulation. Rosiglitazone and GW1929 also suppressed CYP11B2 transactivation. Mutation analysis revealed that the Ad1/CRE element in CYP11B2 5'-flanking region was responsible for the pioglitazone-mediated transrepression. Pioglitazone suppressed ionomycin and a truncated constitutively active form Ca(2+)/calmodulin-dependent kinase I (CaMKI)-mediated CYP11B2 transcriptional activation. A CaMK inhibitor KN-93 attenuated pioglitazone-mediated CYP11B2 transrepression. PPARγ suppresses CYP11B2 expression and aldosterone secretion.

Background Hypertension is a significant global public health problem and recognized as an important risk factor for cardiovascular diseases. This study was designed to assess the current prevalence of hypertension and to explore risk factors associated with hypertension by urban and rural status to guide the prevention and control of hypertension in Jilin province. Methods A multi-stage stratified random cluster sampling method was used to obtain data on hypertension, which was investigated by physical examination and face-to-face questionnaire in July 2014-December 2015. Sample data were analyzed by complex weighted statistical analysis to estimate blood pressure levels and prevalence of hypertension in the province. Multivariable logistic regression analysis was used to identify factors influencing hypertension rates. Results The prevalence of hypertension was significantly higher in rural areas than urban areas (25.93% versus 22.73%, respectively). The rates of hypertension known (46.7% versus 38.1%, respectively), control (13.7% versus 5.0%, respectively), and controlled among treated subjects (38.3% versus 17.5%, respectively) were higher in urban areas than in rural areas (all p < 0.001), while the treatment rate was not statistically significantly different between urban and rural areas (35.9% versus 28.4%, respectively). After adjusting for demographic covariates, hypertension prevalence in rural areas was still significantly greater than in urban areas (adjusted OR = 1.22; 95%CI: 1.10, 1.36; p < 0.001). Common risk factors for hypertension among urban and rural residents included older age; male; married; employed; less education; overweight/obese; greater abdominal waist circumference; family history of hypertension, stroke, or coronary heart disease; current smoker; alcohol consumption; higher visceral adiposity index; and higher body fat percentage. Conclusions This study identified an increased risk for hypertension in rural regions of Jilin province, suggesting that rural hypertension screening and treatment guidelines should receive greater attention.

Objective: To evaluate the clinical effects of thoracoscopy-assisted mini-open surgery for anterior column reconstruction in thoracic spinal tuberculosis. Methods: Fifty-eight patients, 35 men and 23 women, aged 39.2 (range, 19–60) years with thoracic spinal tuberculosis with an average kyphotic angle of 29.2° (range, 18°–42°) underwent thoracoscopy-assisted mini-open surgeries, including thorough debridement and anterior spinal reconstruction. According to the Frankel Grading system, preoperative neurological function was judged as Grade B in 3 cases, Grade C in 7, Grade D in 28, and Grade E in 20. All patients were followed up for an average of 4.6 years. Outcomes were evaluated retrospectively. Results: Surgery was accomplished successfully in all cases. The average operation time was 230 min (range, 180–320 min), the average intraoperative blood loss 570 ml (range, 350–1200 ml), and the mean drainage duration 3.6 days (3–5 days). Complications occurred in 19 patients (32.8%). Neurological improvement of one to three grades had occurred in 29 patients by final follow-up. The average correction rate of the kyphotic angle was 36.4%, and no obvious correction loss was detected during follow-up. No recurrent tuberculosis was found in the group. Conclusions: Thoracoscopy-assisted mini-open surgery provides a simple, safe, effective, and practical technology with minimal invasiveness for the treatment of thoracic spinal tuberculosis.

Age-related morphologic development of human adrenal zona reticularis (ZR) has not been well examined. Therefore, in this study, 44 human young adrenal autopsy specimens retrieved from large archival files (n=252) were examined for immunohistochemical and morphometric analyses. Results demonstrated that ZR became discernible around 4 years of age, and both thickness and ratio per total cortex of ZR increased in an age-dependent fashion thereafter, although there was no significant increment in total thickness of developing adrenal cortex. We further evaluated immunoreactivity of both KI67 and BCL2 in order to clarify the equilibrium between cell proliferation and apoptosis in the homeostasis of developing human adrenals. Results demonstrated that proliferative adrenocortical cells were predominantly detected in the zona glomerulosa and partly in outer zona fasciculata (ZF) before 4 years of age and in ZR after 4 years of age, but the number of these cells markedly decreased around 20 years of age. The number of BCL2-positive cells increased in ZR and decreased in ZF during development. Adrenal androgen synthesizing type 5 17β-hydroxysteroid dehydrogenase (HSD17B5 or AKR1C3 as listed in the Hugo Database) was almost confined to ZR of human adrenals throughout development. HSD17B5 immunoreactivity in ZR became discernible and increased from around 9 years of age. Results of our present study support the theory of age-dependent adrenocortical cell migration and also indicated that ZR development is not only associated with adrenarche, but may play important roles in an initiation of puberty.

Mrf4 is a basic helix-loop-helix (bHLH) transcription factor associated with myogenesis. Two mrf4 transcripts, mrf4_tv1 and mrf4_tv2, were identified in zebrafish generated by alternative splicing. To study their biological functions, we separately injected the Mrf4-morpholinos, including MO1 (mrf4_tv1:mrf4_tv2 knockdown), MO2+MO3 (mrf4_tv1:mrf4_tv2 knockdown), MO3 (mrf4_tv1 knockdown), and MO4 (mrf4_tv2 knockdown), into zebrafish embryos to observe mrf4 gene knockdown phenotypes. No phenotypic abnormalities were observed following injection with 0.5 ng of MO1 but those injected with 4.5, 9, or 13.5 ng displayed curved-body phenotypes, such as indistinct somite boundaries, and a lack of uniformly sized cell blocks. Similar results were also observed in the (MO2+MO3)-, MO3-, and MO4-injected groups. To further investigate the molecular mechanisms that lead to curved-body phenotypes, we stained embryos with α-bungrotoxin and specific monoclonal antibodies F59, Znp1, and Zn5 to detect morphological changes in acetyl-choline receptor (AChR) clusters, muscle fibers, common path of the primary neurons, and secondary neurons axonal projections, respectively. Our results show that the muscle fibers of mrf4_(tv1:tv2)-morphant aligned disorderly and lost their integrity and attachment, while the defects became milder in either mrf4_tv1-morphant or mrf4_tv2-morphant. On the other hand, reduced axonal projections and AChR clusters were found in both mrf4_tv2-morphant and mrf4_(tv1:tv2)-morphant but distributed normally in the mrf4_tv1-morphant. We conclude that Mrf4_tv2 is involved in alignment of muscle fibers, and Mrf4_tv1 might have cooperative function with Mrf4_tv2 in muscle fiber alignment, without affecting the muscle-nerve connection. Developmental Dynamics 237:1043–1050, 2008. © 2008 Wiley-Liss, Inc.

To compare the clinical efficacy in the treatment of post-stroke shoulder-hand syndrome between floating-needle therapy and conventional acupuncture on the basis of rehabilitation training.One hundred cases of post-stroke shoulder-hand syndrome were randomized into a floating-needle group and an acupuncture group, 50 cases in each one. The passive and positive rehabilitation training was adopted in the two groups. Additionally, in the floating-needle group, the floating-needle therapy was used. The needle was inserted at the site 5 to 10 cm away from myofasical trigger point (MTrP), manipulated and scattered subcutaneously, for 2 min continuously. In the acupuncture group, the conventional acupuncture was applied at Jianqian (EX-UE), Jianyu (LI 15), Jianliao (TE 14), etc. The treatment was given once every two days, 3 times a week, and 14 days of treatment were required. The shoulder hand syndrome scale (SHSS), the short form McGill pain scale (SF-MPQ) and the modified Fugl-Meyer motor function scale (FMA) were used to evaluate the damage severity, pain and motor function of the upper limbs before and after treatment in the two groups. The clinical efficacy was compared between the two groups.SHSS score, SF-MPQ score and FMA score were improved significantly after treatment in the two groups (all P0.01), and the improvements in the floating-needle group were superior to those in the acupuncture group (all P0.05). The total effective rate was 94.0% (47/50) in the floating-needle group, which was better than 90.0% (45/50) in the acupuncture group (P0.05).The floating-needle therapy combined with rehabilitation training achieves a satisfactory efficacy on post-stroke shoulder-hand syndrome, which is better than the combined therapy of conventional acupuncture and rehabilitation training.