A Method of Quick and Safe Coal Uncovering by Hydraulic Fracturing in a Multibranch Radial Hole with a Coalbed Methane Well.

Coal and gas outburst accidents occur most frequently during coal uncovering operation, and uncovering-induced outburst accidents have caused great economic losses and casualties. In order to quickly and safely uncover the outburst coal seam at the air intake shaft in panel 2, Zhaozhuang Coal Mine has come up with the technology of multibranch radial borehole hydraulic fracturing (MRBHF) coal uncovering technology in the surface coalbed methane (CBM) well. By conducting fracturing of constructed radial boreholes in the CBM well, the technology increases fractures, enhancing coal permeability and raising gas drainage volume in the uncovering area. It brings the following three major benefits: (1) uncovering coal efficiently, rapidly, and safely; (2) greatly reducing engineering quantity, coal uncovering period, and construction cost, i.e., significantly improving the economic benefits; and (3) efficiently reusing the CBM well. Compared with conventional uncovering measures, the proposed technology shortens the uncovering period by 118 d, enhances the coal seam permeability by 3.55 times, raises the gas drainage rate by 22%, increases the gas drainage volume by 1.93 times, and reduces the engineering volume of drilling by 12,000 m. The safe, rapid, economical, and applicable technology provides scientific guidance and reference for domestic and foreign shaft coal uncovering engineering and possesses important economic and social values and application prospect for safe and efficient production of coal mines.

Research Advances of Prevention and Control of Hydrogen Sulfide in Coal Mines.

Sudden emission and casualty accidents caused by abnormal enrichment of hydrogen sulfide (H2S) in coal mines are becoming frequent increasingly, causing major casualties and environmental pollution. Scholars in various countries have developed various measuring devices for hydrogen sulfide content in coal and rock formations and their calculation methods. The existing prevention and control technologies of H2S in coal mines were summarized in various countries. According to the distribution characteristics, occurrence modes, and emission forms of H2S in coal mines, the prevention and control technologies of H2S in coal-bearing strata, airflow in tunnel, and underground water body are mainly introduced. Analyzed the effects of different ventilation systems on prevention and control of H2S, which include conventional ventilation system, partial homotropal ventilation system, and full homotropal ventilation system. The methods used mainly include neutralization by injecting alkalizer through drilling in coal seams with high pressure, spraying alkalizer in tunnel, attenuation by increasing wind amount, changing the ventilation method, pumping, dredging, and blocking the water that contains H2S as well as comprehensive prevention and control method. The basic agents adopted mainly include sodium carbonate (the mass percentage concentration is about 0.5% ~ 3.0%) and sodium bicarbonate solution, and some basic solution is added by an additive, such as surfactant, Fenton reagent, sodium dodecyl benzene sulfonate, sodium hypochlorite, or chloramine-T. The treatment effect and the main problems of each prevention and control technology are analyzed, and a comprehensive method of prevention and control techniques of H2S in coal mines is proposed. According to current technological level as well as the cost, the effective prevention and control techniques of H2S should take the occurrence, distribution, and emission forms of H2S in coal mines as well as the content into consideration.

Facile Preparation and Self-Assembly of Monodisperse Polystyrene Nanospheres for Photonic Crystals.

We have developed boiling emulsifier-free emulsion polymerization recipes for the synthesis of monodisperse polystyrene (PS) nanospheres with diameters between ca. 100 and 300 nm. The morphologies of the nanospheres during growth were characterized and the results showed that the PS nanospheres with uniform structures could be synthesized rapidly by modifying the reaction conditions. These nanospheres readily self-assemble into three-dimensionally colloidal photonic crystal film and whose photonic band-stop could be tuned over the entire visible spectral region by altering the sphere diameters.

Electrically configurable graphene field-effect transistors with a graded-potential gate.

A device architecture for electrically configurable graphene field-effect transistor (GFET) using a graded-potential gate is present. The gating scheme enables a linearly varying electric field that modulates the electronic structure of graphene and causes a continuous shift of the Dirac points along the channel of GFET. This spatially varying electrostatic modulation produces a pseudobandgap observed as a suppressed conductance of graphene within a controllable energy range. By tuning the electrical gradient of the gate, a GFET device is reversibly transformed between ambipolar and n- and p-type unipolar characteristics. We further demonstrate an electrically programmable complementary inverter, showing the extensibility of the proposed architecture in constructing logic devices based on graphene and other Dirac materials. The electrical configurable GFET might be explored for novel functionalities in smart electronics.

Enhanced Field Emission from a Carbon Nanotube Array Coated with a Hexagonal Boron Nitride Thin Film.

A high-quality field emission electron source made of a highly ordered array of carbon nanotubes (CNTs) coated with a thin film of hexagonal boron nitride (h-BN) is fabricated using a simple and scalable method. This method offers the benefit of reproducibility, as well as the simplicity, safety, and low cost inherent in using B(2)O(3) as the boron precursor. Results measured using h-BN-coated CNT arrays are compared with uncoated control arrays. The optimal thickness of the h-BN film is found to be 3 nm. As a result of the incorporation of h-BN, the turn-on field is found to decrease from 4.11 to 1.36 V µm(-1), which can be explained by the significantly lower emission barrier that is achieved due to the negative electron affinity of h-BN. Meanwhile, the total emission current is observed to increase from 1.6 to 3.7 mA, due to a mechanism that limits the self-current of any individual emitting tip. This phenomenon also leads to improved emission stability and uniformity. In addition, the lifetime of the arrays is improved as well. The h-BN-coated CNT array-based field emitters proposed in this work may open new paths for the development of future high-performance vacuum electronic devices.

Graphene nanomesh photodetector with effective charge tunnelling from quantum dots.

Graphene nanomesh (GNM)-based optoelectronics integrated with quantum dots (QDs) are investigated in this article. The charge transfer mechanism in the QDs/GNM interface is probed in four terminal gated FET-type photodetectors. The insulating ligand is used to make the GNM/ligand/QDs vertically behave like a metal/insulate/semiconductor (MIS) structure to facilitate the charge tunnelling. With the current constraint effect of the GNM and the effective charge tunnelling, a high-performance photodetector is fabricated with higher responsivity, higher on/off ratio and shorter response time. The results of our analysis and experimental approach can be extended to future graphene-based photodetectors, as long as suitable ligands and an effective architecture are chosen for this type of device.

High-performance Ge quantum dot decorated graphene/zinc-oxide heterostructure infrared photodetector.

A novel size-controllable germanium quantum dot (Ge QD) is synthesized and decorated onto reduced graphene oxide (RGO) fragments to overcome the low infrared (IR) photoresponses (∼0.1 A/W)13,14 of pristine graphene. With the integration of flexible substrate, monolayer graphene (MLG) electrode and n-type zinc oxide (ZnO), a high-performance QD-decorated-RGO/ZnO heterostructure infrared photodetector is reported in this study. The Ge QD-decorated-RGO hybrid photosensitive composite improves the responsivity (∼9.7 A/W, 1400 nm) in IR waveband without sacrificing the response speed (∼40 µs rise time and 90 µs recovery time). In addition, the effective barrier formed between graphene and ZnO interface restricts the dark current (∼1.4 nA, -3 V) to guarantee the relatively excellent rectifying behavior and high on/off ratio (∼10(3)) for this IR photodetector. With these superior inherent properties and micron-sized sensing active area, this photodetector manifests great potential in the future application of graphene-based IR photodetector.

An isogenetic myoblast expression screen identifies DUX4-mediated FSHD-associated molecular pathologies.

Facioscapulohumeral muscular dystrophy (FSHD) is caused by an unusual deletion with neomorphic activity. This deletion derepresses genes in cis; however which candidate gene causes the FSHD phenotype, and through what mechanism, is unknown. We describe a novel genetic tool, inducible cassette exchange, enabling rapid generation of isogenetically modified cells with conditional and variable transgene expression. We compare the effects of expressing variable levels of each FSHD candidate gene on myoblasts. This screen identified only one gene with overt toxicity: DUX4 (double homeobox, chromosome 4), a protein with two homeodomains, each similar in sequence to Pax3 and Pax7. DUX4 expression recapitulates key features of the FSHD molecular phenotype, including repression of MyoD and its target genes, diminished myogenic differentiation, repression of glutathione redox pathway components, and sensitivity to oxidative stress. We further demonstrate competition between DUX4 and Pax3/Pax7: when either Pax3 or Pax7 is expressed at high levels, DUX4 is no longer toxic. We propose a hypothesis for FSHD in which DUX4 expression interferes with Pax7 in satellite cells, and inappropriately regulates Pax targets, including myogenic regulatory factors, during regeneration.

Catalase overexpression does not impair extensor digitorum longus muscle function in normal mice.

Catalase is a major antioxidant enzyme. Increasing catalase expression represents a promising avenue to improve muscle function in certain physiological conditions and in some muscle diseases. We hypothesized that catalase overexpression should not impair normal muscle contraction. We delivered a hemagglutinin (HA)-tagged human catalase gene to normal mouse muscle by an adeno-associated viral vector (AAV). Western blot and immunostaining revealed efficient expression of HA-tagged catalase. Enzymatic assay demonstrated an approximately threefold increase in catalase activity in AAV-infected muscles. Catalase overexpression impaired neither twitch nor tetanic tension in the extensor digitorum longus (EDL) muscle. Furthermore, EDL fatigue response was not altered. Taken together, we have developed a novel AAV vector to enhance catalase expression. Lack of apparent toxicity in normal muscle strongly supports further exploration of this vector to reduce oxidative stress-induced muscle damage.

Synthetic intron improves transduction efficiency of trans-splicing adeno-associated viral vectors.

Trans-splicing adeno-associated viral (AAV) vectors hold great promise in many gene therapy applications. We have shown that rational selection of the gene-splitting site in a therapeutic target gene can lead to extremely efficient trans-splicing vectors [Lai, Y., Yue, Y., Liu, M., Ghosh, A., Engelhardt, J.F., Chamberlain, J.S., and Duan, D. (2005). Nat. Biotechnol. 23, 1435-1439]. Our original strategy requires the screening of endogenous introns that are capable of overcoming the mRNA accumulation barrier. To further develop transsplicing vectors, we have tested whether the use of a generic synthetic intron can bypass the labor-intensive intron-screening process. Two previously characterized exon/intron/exon junctions (60/60/61 and 63/63/64, respectively) in the 6 kb minidystrophin gene were used as templates to represent highly efficient (60/60/61) and relatively poor (63/63/64) gene-splitting sites. We compared RNA production from the reconstituted viral genome and transduction efficiency of the trans-splicing vectors in dystrophin-null mdx mouse skeletal muscle. Our results suggest that a synthetic intron can successfully overcome the mRNA accumulation barrier at the exon 63/64 junction. Furthermore, when the gene was split at the exon 63/64 junction, the synthetic intronbased vectors performed better than the endogenous intron-based vectors. When the gene was split at the exon 60/61 junction, we observed only nominal improvement in mRNA production. Nevertheless, vectors based on the exon 60/61 junction remain the best set in transduction efficiency. Taken together, our results suggest that optimizing intron sequence may boost the transduction efficiency of trans-splicing AAV vectors.

C-terminal-truncated microdystrophin recruits dystrobrevin and syntrophin to the dystrophin-associated glycoprotein complex and reduces muscular dystrophy in symptomatic utrophin/dystrophin double-knockout mice.

C-terminal-truncated (DeltaC) microdystrophin is being developed for Duchenne muscular dystrophy gene therapy. Encouraging results have been achieved in the mdx mouse model. Unfortunately, mdx mice do not display the same phenotype as human patients. Evaluating DeltaC microdystrophin in a symptomatic model will be of significant relevance to human trials. Utrophin/dystrophin double-knockout (u-dko) mice were developed to model severe dystrophic changes in human patients. In this study we evaluated the therapeutic effect of the DeltaR4-R23/DeltaC microdystrophin gene (DeltaR4/DeltaC) after serotype-6 adeno-associated virus-mediated gene transfer in neonatal u-dko muscle. At 2 months after gene transfer, the percentage of centrally nucleated myofiber was reduced from 89.2 to 3.4% and muscle weight was normalized. Furthermore, we have demonstrated for the first time that DeltaC microdystrophin can eliminate interstitial fibrosis and macrophage infiltration and restore dystrobrevin and syntrophin to the dystrophin-associated glycoprotein complex. Interestingly neuronal nitric oxide synthase was not restored. The most impressive results were achieved in muscle force measurement. Neonatal gene therapy increased twitch- and tetanic-specific force. It also brought the response to eccentric contraction-induced injury to the normal range. In summary, our results suggest that the DeltaR4/DeltaC microgene holds great promise in preventing muscular dystrophy.

Efficient in vivo gene expression by trans-splicing adeno-associated viral vectors.

Although adeno-associated virus (AAV)-mediated gene therapy has been hindered by the small viral packaging capacity of the vector, trans-splicing AAV vectors are able to package twice the size of the vector genome. Unfortunately, the efficiency of current trans-splicing vectors is very low. Here we show that rational design of the gene splitting site has a profound influence on trans-splicing vector-mediated gene expression. Using mRNA accumulation as a guide, we generated a set of efficient trans-splicing vectors and achieved widespread expression of the 6-kb DeltaH2-R19 mini-dystrophin gene in skeletal muscle of mdx mice, a model for Duchenne muscular dystrophy. The dystrophic phenotype was ameliorated in both adult and aged mice. This demonstrates the use of trans-splicing vectors to efficiently express a large therapeutic structural protein. This strategy should be applicable to other large therapeutic genes or large transcription regulatory elements.

Adeno-associated virus-mediated microdystrophin expression protects young mdx muscle from contraction-induced injury.

Duchenne muscular dystrophy (DMD) is the most common inherited lethal muscle degenerative disease. Currently there is no cure. Highly abbreviated microdystrophin cDNAs were developed recently for adeno-associated virus (AAV)-mediated DMD gene therapy. Among these, a C-terminal-truncated DeltaR4-R23/DeltaC microgene (DeltaR4/DeltaC) has been considered as a very promising therapeutic candidate gene. In this study, we packaged a CMV.DeltaR4/DeltaC cassette in AAV-5 and evaluated the transduction and muscle contractile profiles in the extensor digitorum longus muscles of young (7-week-old) and adult (9-month-old) mdx mice. At approximately 3 months post-gene transfer, 50-60% of the total myofibers were transduced in young mdx muscle and the percentage of centrally nucleated myofibers was reduced from approximately 70% in untreated mdx muscle to approximately 22% in microdystrophin-treated muscle. Importantly, this level of transduction protected mdx muscle from eccentric contraction-induced damage. In contrast, adult mdx muscle was more resistant to AAV-5 transduction, as only approximately 30% of the myofibers were transduced at 3 months postinfection. This transduction yielded marginal protection against eccentric contraction-induced injury. The extent of central nucleation was also more difficult to reverse in adult mdx muscle (from approximately 83% in untreated to approximately 58% in treated). Finally, we determined that the DeltaR4/DeltaC microdystrophin did not significantly alter the expression pattern of the endogenous full-length dystrophin in normal muscle. Neither did it have any adverse effects on normal muscle morphology or contractility. Taken together, our results suggest that AAV-mediated DeltaR4/DeltaC microdystrophin expression represents a promising approach to rescue muscular dystrophy in young mdx skeletal muscle.

Full-length dystrophin expression in half of the heart cells ameliorates beta-isoproterenol-induced cardiomyopathy in mdx mice.

Gene therapy holds great promise for curing Duchenne muscular dystrophy (DMD), the most common fatal inherited childhood muscle disease. Success of DMD gene therapy depends upon functional improvement in both skeletal and cardiac muscle. Numerous gene transfer studies have been performed to correct skeletal muscle pathology, yet little is known about cardiomyopathy gene therapy. Since complete transduction of the entire heart is an impractical goal, it becomes critical to determine the minimal level of correction needed for successful DMD cardiomyopathy gene therapy. To address this question, we generated heterozygous mice that persistently expressed the full-length dystrophin gene in 50% of the cardiomyocytes of mdx mice, a model for DMD. We questioned whether dystrophin expression in half of the heart cells was sufficient to prevent stress-induced cardiomyopathy. Heart function of mdx mouse is normal in the absence of external stress. To determine the therapeutic effect, we challenged 3-month-old mice with beta-isoproterenol. Cardiomyocyte sarcolemma integrity was significantly impaired in mdx but not in heterozygous and C57Bl/10 mice. Importantly, in vivo closed-chest hemodynamic assays revealed normal left ventricular function in beta-isoproterenol-stimulated heterozygous mice. Since the expression profile in the heterozygous mice mimicked viral transduction, we conclude that gene therapy correction in 50% of the heart cells may be sufficient to treat cardiomyopathy in mdx mice. This finding may also apply to the gene therapy of other inherited cardiomyopathies.

The effects of verbascoside on plasma lipid peroxidation level and erythrocyte membrane fluidity during immobilization in rabbits: a time course study.

We studied the changes in the level of plasma lipid peroxidation indicated by malondialdehyde (MDA) level and erythrocyte membrane fluidity expressed by the value of order parameter (S) during single hindlimb immobilization of 21 days using thiobarbituric acid - reactive substances method and spin label electron spin resonance method, respectively. The impacts of verbascoside that has been proved its antioxidative activity on the measured parameters were examined. 11 New Zealand white rabbits were immobilized and divided into two groups. The rabbits in the verbascoside group were administrated with 0.8 mg/kg of verbascoside twice a day orally throughout the immobilization. The rabbits in the placebo group were treated with normal saline. In placebo group, the results showed that the level of MDA significantly increased on day 3, peaked on day 7, and was still significantly higher on day 14 of immobilization, compared with the value measured before immobilization. The value of S reached the highest on day 7 and subsequently lowered but still higher on day 14 than those measured before immobilization. Compared with placebo group, there were lower MDA level (P < 0.05, 0.001, and 0.05 for days 3, 7, and 14, respectively) and higher erythrocyte membrane fluidity (P < 0.05, 0.001, and 0.001 for days 3, 7, and 14, respectively) in verbascoside group. The data indicated that immobilization caused temporal changes of increase in plasma lipid peroxidation and decrease in erythrocyte membrane fluidity. Verbascoside might have the effects to moderate oxidative stress and erythrocyte membrane fluidity during immobilization.

Immobilization-induced changes in erythrocyte membrane fluidity in rabbits: a spin-label electron spin resonance study.

It has been shown that many diseases are linked to abnormalities of the erythrocyte membrane. This study observed the changes in erythrocyte membrane fluidity during an immobilization period of 21 days. The right hindlimbs of male adult New Zealand white rabbits were immobilized for 21 days. Blood samples were collected and heparinized prior to immobilization and on days 1, 3, 7, 14, and 21 of immobilization. The membrane fluidity of erythrocytes was measured by spin-label electron spin resonance (ESR). The membrane fluidity was expressed by the value of order parameter (S). The results showed a significant increase of S on days 7 and 14 of immobilization (p <0.01). The highest value of S was found on day 7 of immobilization; thereafter, S gradually declined. Compared with the value measured before immobilization, there was no significant difference in the value of S on day 21. The results show that immobilization significantly affected erythrocyte membrane fluidity, indicating a systemic and temporal response ofthe immobilized body. This study is the first to document the time-course of changes in erythrocyte membrane fluidity in rabbits with single hindlimb immobilization during 21 days. These results may be helpful in studying the pathophysiology of immobilization.