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This study aimed to create prognostic signatures to predict AML patients' survival using alternative splicing (AS) events. The AS data, RNA sequencing data, and the survival statistics of 136 AML patients were obtained from The Cancer Genome Atlas (TCGA) and TCGA SpliceSeq databases. Total 34,984 AS events generated from 8,656 genes, 2,583 of which were survival-associated AS events, were identified using univariate Cox regression. The prognostic models constructed using independent survival-associated AS events revealed that low-risk splicing better predicted patients' survival. ROC analysis indicated that the predictive efficacy of the alternate terminator model was best in the area under the curve at 0.781. Enrichment analysis revealed several important genes (TP53, BCL2, AURKB, PPP2R1B, FOS, and BIRC5) and pathways, such as the protein processing pathway in the endoplasmic reticulum, RNA transport pathway, and HTLV-I infection pathway. The splicing network of splicing events and factors revealed interesting interactions, such as the positive correlation between HNRNPH3 and CALHM2-13010-AT, which may indicate the potential splicing regulatory mechanism. Taken together, survival-associated splicing events and the prognostic signatures for predicting survival can help provide an overview of splicing in AML patients and facilitate clinical practice. The splicing regulatory network may improve the understanding of spliceosomes in AML.

The accumulation and spatial distribution of economically important petroleum in sedimentary basins are primarily controlled by its migration from source rocks through permeable carrier beds to reservoirs. Tracing petroleum migration entails the use of molecular indices established according to sorption capacities of polar molecules in migrating petroleum. However, little is known about molecular sorption capacities in natural migration systems, rendering these indices unreliable. Here, we present a new approach based on a novel concept of relative sorption coefficient for quantitatively assessing sorption capacities of polar molecules during natural petroleum migration. Using this approach, we discovered previously unrecognized “stripping” and “impeding” effects that significantly reduce the sorption capacities of polar compounds. These discoveries provide new insights into the behaviors of polar compounds and can easily explain why traditional molecular indices yield incorrect information about petroleum migration. In light of these new findings, we established new molecular indices for tracing petroleum migration. We demonstrate via case studies that the newly established indices, unlike traditional molecular indices, are reliable and effective in tracing petroleum migration. Our approach can be applied to diverse basins around the world to reveal distribution patterns of petroleum, which would decrease environmental risks of exploration by reducing unsuccessful wells.

Al2TiO5/TiO2/ZrO2 plasma electrolytic oxidation coating deposited on the surface of Ti–6Al–4V alloy was prepared in NaAlO2–NaH2PO2–NaOH electrolytes system, which was doped with ZrO2 micro-particles by a kind of pulsed bipolar plasma electrolytic oxidation. The microstructure, surface morphology and phase composition of the Al2TiO5/TiO2/ZrO2 coatings were studied by SEM and XRD, respectively. Its cyclic oxidation behaviour was studied over the temperature of 1000°C for 100 hours in dry air. After high-temperature oxidation, the original crater eruption topography of plasma electrolytic oxidation coating has disappeared and it obviously took a decreasing tendency of both pore numbers and their diameters. The scales of original plasma electrolytic oxidation coating consisted mostly of Al2TiO5, R-TiO2 and α-Al2O3intermixed with small amounts of ZrO2 particles. They were transferred into R-TiO2, α-Al2O3 and ZrO2 with small amounts of Al2TiO5 after high-temperature oxidation procedure. The weight gains of plas...

The isothermal oxidation behaviours of Al2TiO5/TiO2/ZrO2 composite coating on titanium alloy (TC4) deposited by micro-arc oxidation technique were studied at different temperature points of 1073, 1173 and 1273 K in dry air. The surface morphology and phase composition of all coated samples were investigated before and after oxidation by SEM and XRD, respectively. The weight gaining curves of the coated samples were plotted and the high temperature oxidation rates were evaluated, showing that the oxidation kinetics followed the parabolic rate law. And with increasing oxidation temperature, their oxidation rates also increased. The original phases of Al2TiO5/TiO2/ZrO2 coating on TC4 alloy consisted mostly of Al2TiO5 and R-TiO2 intermixed with small amounts of ZrO2 and the phased changed into mainly of R-TiO2, Al2O3 and little Al2TiO5 when after 1273 K oxidation for 1 h, whose average parabolic rate constant was 6.43 × 10−13 g2 cm−4 s−1, greatly lower than that of blank samples, 6.01 × 10−12 g2 cm−4 ...

The objective of this study was to assess the efficacy of exposing Rosenmüller valve during endonasal endoscopic dacryocystorhinostomy (EES-DCR) in treating refractory nasolacrimal duct obstruction.Thirty six patients (47 eyes) with refractory nasolacrimal duct obstruction were entered into this study, including 13 patients (15 eyes) underwent nasal lacrimal duct intubation before, 18 patients (26 eyes) with small lacrimal sac, 2 patients underwent DCR before, 1 patient with functional nasal lacrimal duct obstruction, 1 patient with lacrimal mucocele, 1 patient with nasal trauma. During the DCR, the bone window was enlarged as wide as possible and the valve of Rosenmüller was exposed. The intubation of the lacrimal apparatus was performed if the lacrimal sac was very small and the Rosenmüller valve was tight. Functional endoscopic sinus surgery and septoplasty were performed simultaneously if necessary. Postoperatively, each patient was assessed by the symptom and nasal endoscopy.The postoperative follow-up period ranged from 3 to 36 months. Thirty two patients(89.4%, 42/47 eyes) were cured, which presented as the disappearance of symptoms, the patency of the lacrimal ostium and the lacrimal duct irrigation was clear. One patient (2.1%, 1/47 eyes) was improved, which presented as the improvement of symptoms, a few hypertrophic scar around the lacrimal ostium and the lacrimal duct irrigation was clear. Four cases failed (8.5%, 4/47), which presented as symptomatic and obstruction of the lacrimal ostium. The total effective rate reached to 91.5%. No serious postoperative complications were found in all cases.To expose Rosenmüller valve during endoscopic DCR in treating patients with refractory nasolacrimal duct obstruction could obviously increase the operative success rate, fewer complications, with no facial surgical scar. It is an ideal way to treat the refractory nasolacrimal duct obstruction.

To investigate the characteristics of children male soccer players' ankle imaging features.From October 2015 to February 2016, a total of 32 male children players of two soccer clubs in Guangzhou were enrolled in this study.The ages of all cases were from 10 to 14 years, average age was (11.3±0.9) years.A total of 15 male children in region ordinary primary and secondary school students were set as control group, ages were from 10 to 14 years, average age was (12.1±1.2) years. All objects' ankle were examined by X-ray for positive and lateral positions; routine CT scanning and then on the workstation restructuring for axial, coronal and sagittal slices; and examined by MR. MR scan was with special surface coil for ankle joint for horizontal axis T2WI; coronal T1WI; coronary proton density weighted imaging (PWI); sagittal T2WI with fat suppression; sagittal PWI with isotropic and fat suppression sequence of fast field echo. The ankle bone morphological structures were observed on X-ray; the ankle bone mineral density, cortical bone thickness and sesamoid bone quantity was being observed and measured on CT; and the tenosynovitis, Achilles tendinitis, synovitis, and cancellous bone edema signal were observed on MR.For study group, a total of 32 cases and 64 ankles joints were completed by X-ray, CT and MR examination.A total of 15 cases and 30 ankles joints were completed by X-ray in control group, 26 ankle joints were completed by CT scan and 22 ankle joints were completed by MR examination.X-ray examination showed there was no statistically significant difference between the two groups in ankle bone structure.CT showed that navicular bone CT value was (296±82) HU in research group and navicular bone CT value was (266±107) HU in control group, the difference was statistically significant (P=0.03). MR showed the incidence of diseases in research group that the tendon sheath peripheral inflammation was 92.2% (59/64), Achilles tendon lesions was 18.8%(12/64), edema of cancellous bone was 73.4% (47/64) , lateral malleous ligaments injuried was 43.8%(28/64), synovitis or effusion in posterior ankle was 87.5% (56/64). The incidence of diseases in control group was that tendon sheath peripheral inflammation was 31.8%(7/22), Achilles tendon lesions was 0/22, edema of cancellous bone was 0/22, lateral malleous ligaments injuried was 0/22, synovitis or effusion in posterior ankle was 54.5% (12/22). There was statistically significant difference between these two groups (all P0.05).Compared with the control group, children's male soccer players ankle bone structure, bone cortex thickness and bone mineral density there were no obvious difference. In the tenosynovitis, Achilles tendinitis, synovitis, bone marrow edema, lateral malleous ligaments injuried were significantly higher than the control group.

Different protocols incorporating methyl-β-cyclodextrin (MBCD) and reduced glutathione (GSH) have been reported to improve IVF recovery of cryopreserved mouse sperm on a C57BL/6 (J and N) genetic background. However, it is not clear which IVF protocol is most appropriate when using the various methods to cryorecover sperm with different sperm quality and sample volumes. Therefore, in the present study we correlated sperm motility with fertilization rate and compared the efficiency of different IVF methods using various sperm samples so as to establish general guidelines for mouse sperm cryorecovery by IVF. High linear correlation between sperm fertilization rate and progressive motility was found, R(2) was 0.9623 and 0.9993 for pre-freezing and post-thaw progressive motility, respectively. High amounts of cryoprotective agent (CPA) were observed to impair both sperm capacitation and fertilization. Moreover, the presence of a large number of immotile sperm in the sperm-oocyte co-incubation drop was found to reduce IVF success which could be partially reversed by supplementation using monothioglycerol (MTG) during centrifugation. It was concluded that the efficiency of IVF using cryorecovered mouse sperm in media containing MBCD and GSH can be predicted from sperm progressive motility. High concentrations of CPA and immotile sperm should be mitigated prior to IVF. The optimum IVF method should be selected based on sperm sample volume and sperm parameters.

Phosphorylation of eIF2α is an important strategy for living organisms to adapt to metabolic and physiological changes that are often associated with external stimuli. GCN2 is one of the well-studied eIF2α kinases in yeast and mammals, which is responsible for the survival of the organism under amino acid starvation. Despite the downstream reactions being quite divergent, AtGCN2 exhibits a high primary sequence similarity to its yeast and animal counterparts. In this study, we provide experimental evidence to show that AtGCN2 shares similar biochemical properties to the yeast and animal homologues. Our in vitro assays demonstrate the binding of the C-terminus of AtGCN2 to uncharged tRNA molecules and the enzymatic activities of AtGCN2 on both eIF2α homologues in A. thaliana, thus providing essential information for further understanding the functions of plant general control non-repressible (GCN) homologues.

The effects of austenitising temperature and temper treatment on the microstructure, hardness and abrasive wear resistance of a high carbon high vanadium high speed steel roll of composition 2·2–2·4C, 5·5–6·5V, 4·5–5·5Cr, 14·0–16·0(W+2Mo), 1·0–1·4Ni and 1·2–1·5Nb (wt-%) were investigated by light optical microscopy, scanning electron microscopy, backscattered electron image, X-ray diffraction and hardness and wear testing. It was found that the hardness increases as the austenitising temperature increases to a maximum of 1050°C, and then begins to decrease because of the increase in retained austenite. Moreover, retained austenite begins to transform into martensite with increasing tempering temperature. The retained austenite in the quenched structure leads to the temper hardening of the rolls, which promotes improved abrasive wear resistance. The roll life of high carbon high vanadium high speed steel compound rolls austenitised at 1050–1080°C and tempered at 540–560°C increased by 400% (single ...

In-doped zinc oxide ( ZnO:In ) thin films with thickness from 157 nm to 592 nm have been deposited on glass substrates by radio frequency (RF) magnetron sputtering. The effect of the film thickness on the structural, electrical and optical properties of ZnO:In thin films has been investigated. It is found that the films are hexagonal wurtzite structure with c-axis perpendicular to the substrate, and with increasing thickness, the crystallinity, the grains size and the conductivity of the films increases, but the strains along c-axis and the transmittance decrease. The decrease of the resistivity in a thicker film is attributed to the slight increase of the carrier concentration and the significant increase of Hall mobility. The transmittance of all the films is over 80% in the visible region (400–800 nm) and the band gap decrease with the increase of film thickness. The film with the thickness of around 303 nm has the resistivity of 6.07 × 10-3 Ω⋅ cm and the transmittance of 90% in the visible range. Based on the good conductivity and high transmittance, the ZnO:In films prepared by magnetron sputtering can be regarded as a potential transparent electrode.

Porous Ni–8YSZ (zirconia stabilised by 8 mol-% yttria) coatings were prepared by electron beam physical vapour deposition (EB–PVD) technique with a high deposition rate of 2 μm min−1 via simultaneously evaporating the ingots of metal Ni and 8YSZ. The coating composition showed a significant dependence on the deposition position in vacuum chamber. The porosity, hardness and electrical conductivity of the coating were severely influenced by Ni content in this coating. The SEM and TEM observations exhibited that the coating presented a distinct columnar structure and many pores were uniformly dispersed at the boundaries of the columns. A Brunauer, Emmet and Teller analysis demonstrated that the average diameter of the pores in this coating was 25 nm and the coating had a high specific surface area of 3·232 m2 g−1.

In order to reduce concentration polarisation from the solid oxide fuel cell (SOFC) anode and the internal stress originating from the thermal expansion mismatch between the anode and electrolyte, a NiO–YSZ coating with a graded composition distribution was prepared by electron beam physical vapour deposition (EB-PVD) through adjusting the electron beam currents. Then the graded Ni–YSZ coating was obtained after a reduction treatment from this NiO–YSZ coating. A graded distribution in contents of both nickel and zirconium was found in this Ni–YSZ coating from the electron probe microanalysis (EPMA) result. Scanning electron microscopy (SEM) observation exhibited a continuous variation in porosity and pore diameter along the direction across the coating thickness. In this coating, a high porosity of up to 43% and an average pore diameter of about 1 µm were obtained in the part close to the substrate, while a low porosity of 10% and mean pore diameters of about tens of nanometres were achieved in th...

A comparison of the California bearing ratio (CBR) for recycled glass (RG) with other recycled materials and a basaltic virgin aggregate, all having similar gradations, revealed that the CBR of RG is superior to that of recycled asphalt pavement but less than that of recycled concrete and virgin aggregate. Direct shear tests were then run on the as-received gradation to derive strength parameters for RG prepared at very high and very low relative densities. For “dense” RG, the peak failure envelope was nonlinear with secant friction angles varying from 50° to 61°. For “loose” RG, the peak and critical state failure envelopes were linear with friction angles of 41° and 38°, respectively. Bolton's postulate that the peak friction angle is approximately equal to the critical friction angle plus 0.8 times the maximum angle of dilation works well for the RG tested in direct shear. A friction angle of 38° at critical state is significant, implying that RG has the potential to be used in even more foundation and ground improvement applications that are so often associated with transportation infrastructure construction. With increased use of recycled materials, civil engineers can help comply with the demand for sustainable development, a major theme in society today.

The measurement and determination of the lithium ion battery's electrochemical impedance spectroscopy (EIS) and the application of EIS to battery classification are researched in this paper. The lithium ion battery gets extensive applications due to its inherent advantages over other batteries. For proper and sustainable performance, it is very necessary to check the uniformity of the lithium ion batteries. In this paper, the equivalent circuit of the lithium ion battery is analyzed; the design of hardware circuit based on DSP and software that calculates the EIS of the lithium ion battery is critically done and evaluated. The parameters of the lithium ion equivalent circuit are determined, the parameter values of li-ion equivalent circuit are achieved by least square method, and the application of Principal Component Analysis (CPA) to the battery classification is analyzed.

Telluride-bearing gold deposits of the Pingyi area, western Shandong, China, are located on the southeastern margin of the North China Craton. There are two main types of deposits: (i) mineralized cryptoexplosive breccia, e.g., Guilaizhuang; and (ii) stratified, finely-disseminated mineralization hosted in carbonate rocks, e.g., Lifanggou and Mofanggou deposits. In Guilaizhuang, the cryptoexplosive breccia is formed within rocks of the Tongshi complex and Ordovician dolomite. The mineralization is controlled by an E–W-trending listric fault. Stratified orebodies of the Lifanggou and Mofanggou deposits are placed along a NE-trending, secondary detachment zone. They are hosted within dolomitic limestone, micrite and dolomite of the Early-Middle Cambrian Changqing Group. The mineralization in the ore districts is considered to be related to the Early Jurassic Tongshi magmatic complex that formed in a continental arc setting on the margin of the North China Craton. The host rocks are porphyritic and consist predominantly of medium- to fine-grained diorite and pyroxene (hornblende)-bearing monzonite. SHRIMP U–Pb zircon dating of diorites give a 206Pb/238U weighted mean age of 175.7 ± 3.8 Ma. This is interpreted as representing the crystallization age of the Tongshi magmatic complex. Considering the contact relationships between the magmatic and host sedimentary rocks, as well as the genetic link with the deposits, we conclude that this age is relevant also for the formation of mineralization in the Pingyi area. We hence consider that the deposits formed in the Jurassic. The principal gold minerals are native gold, electrum and calaverite. Wall-rock alteration comprises pyritization, fluoritization, silicification, carbonatization and chloritization. Fluid inclusion studies indicate that all the analyzed inclusions are of two-phase vapor–liquid NaCl–H2O type. Homogenization temperatures of the fluid inclusions vary from 103 °C to 250 °C, and the ice melting temperatures range from −2.5 °C to −13.5 °C, corresponding to a salinity range of 4.65 to 17.26 wt.% NaCl equiv. The δ34S values of pyrite associated with gold mineralization exhibit a narrow range of −0.71 to + 2.99‰, implying that the sulfur was probably derived from the mantle and/or dioritic magma. The δ13CPDB values of the fluid inclusions in calcite range from −7.3 to 0.0‰. The δ18OSMOW values of vein quartz and calcite range from 11.5 to 21.5‰, corresponding to δ18Ofluid values of −1.1 to 10.9‰; δD values of the fluid inclusions vary between −70 and −48‰. The isotope data for all three deposits suggest mixing of ore-forming fluids derived from the mantle and/or magma with different types of fluids at shallow levels. Pressure release and boiling of the fluids, as well as fluid-rock interaction (Lifanggou and Mofanggou) and mixing of magmatically-derived fluids with meteoritic waters (Guilaizhuang) played an important role in the ore-forming processes.

This letter presents a V-band direct injection-locked frequency divider (ILFD) using forward body bias technology. The divider is implemented in a 0.13 μm CMOS process. The measurements show that the free-running frequency of the divider is 28.67 GHz, the total locking range is 16.2% at 58 GHz with 3.93 mW from a low supply voltage of 0.7 V, and the measured phase noise of the divider is -123.5 dBc/Hz at 500 KHz offset. The output power of the divider is higher than -9 dBm from 54 to 61 GHz, and a good figure of merit is achieved.

Silkworm (Bombyx mori L.) is one of the important economic insects. Silkworm rearing and silk industry plays an important role in China, India and other developing countries. In the long history of sericultural practice, introduction of silkworm strains with high resistance to diseases has greatly improved cocoon and silk quality and productivity. However, current silkworm breeding is mainly based on traditional method that involves high input of time and labour. In order to increase the selection efficiency and accuracy for future silkworm breeding, it is necessary to establish a molecular marker-assisted selection system. In our study, three silkworm near isogenic lines that had different resistance to nuclear polyhedrosis virus (NPV) were established by means of different hybridization methods. A total of 150 random amplified polymorphic DNA (RAPD) random primers were used to screen molecular markers. Among them, two molecular markers OPA-18700 and OPY-11400 were found linked to major genes resistant and susceptible to NPV, respectively. Validity of the molecular markers was proved in F2 populations.

Phosphorylation of eIF2α is an important strategy for living organisms to adapt to metabolic and physiological changes that are often associated with external stimuli. GCN2 is one of the well-studied eIF2α kinases in yeast and mammals, which is responsible for the survival of the organism under amino acid starvation. Despite the downstream reactions being quite divergent, AtGCN2 exhibits a high primary sequence similarity to its yeast and animal counterparts. In this study, we provide experimental evidence to show that AtGCN2 shares similar biochemical properties to the yeast and animal homologues. Our in vitro assays demonstrate the binding of the C-terminus of AtGCN2 to uncharged tRNA molecules and the enzymatic activities of AtGCN2 on both eIF2α homologues in A. thaliana, thus providing essential information for further understanding the functions of plant general control non-repressible (GCN) homologues.

ICSI (intracytoplasmic sperm injection) is an important and commonly used assisted reproductive technology (ART) in humans and animals. However, conventional ICSI has proven to be difficult and inefficient, especially in mice. Fortunately, the application of piezo-electric technology to assist the injection process has made ICSI more effective and efficient in recent years. Despite this advance, the pipette tip is unstable in response to the force exerted by the piezoelectric pulse during injection. To some extent, this instability can be mitigated by the use of mercury, which is highly toxic. In an effort to overcome these hurdles, we developed and tested a completely different and mercury-free technology, called the “Ros-Drill©” (rotationally oscillating drill), which uses a microprocessor-controlled actuator to generate rotational oscillations on a spiked micropipette. ICSI utilizing the Ros-Drill© requires neither mercury nor piezoelectricity. Early experimental trials reveal high survival (≫70% of the injected oocytes), fertilization (≫ 80% of the survived oocytes), and blastocyst formation rates (∼ 50% of the survived oocytes). After surgical transfer of blastocysts created by Ros-Drill© ICSI into pseudopregnant surrogate mothers, healthy pups were born and weaned. Because the Ros-Drill ICSI technique is automated, a very short training period is required, especially for specialists. These studies in mice have revealed, that Ros-Drill ICSI has mentioned advantages over conventional and piezo-assisted ICSI starting with the elimination of mercury in the process.

Extracellular signal-regulated kinases (ERK) such as ERK1 [p44 mitogen-activated protein kinase (MAPK)] and ERK2 (p42 MAPK) are activated in the central nervous system under physiological and pathological conditions such as ischemia and epilepsy. Our aim is to investigate ERK1, ERK2, and phosphorylated ERK (p-ERK) (Thr202/ Tyr 204) expression in the temporal lobe of patients with intractable epilepsy (IE) and to explore its possible role of ERK in it. Tissue samples from temporal neocortices of 40 patients who had surgery for IE were used to detect ERK1, ERK2, and p-ERK (Thr 202/Tyr 204) expression through immunohistochemistry and western blot. We compared these tissues against 17 histological normal temporal lobes from head-traumat patients. ERK1, ERK2, and p-ERK in IE were significantly higher than those in the controls. They were mainly expressed in the cytoplasm of neurons and glial cells. There was also increased detection of p-ERK in the gliotic cortex of IE compared with the non-gliotic cortex. These findings were consistently observed in western blot and immunohistochemistry techniques. ERK expression in patients with IE was significantly increased compared with the controls. This suggested a probable role of ERK in the pathogenesis of IE.

Recordings were obtained from a total of 25 normal subjects of the electromyographic (EMG) responses in the sternocleidomastoid muscle (SCM) to intense sound stimuli. While previous authors have demonstrated these responses exist, it has remained unclear whether the EMG response is unilateral or bilateral in nature. Accordingly, we chose a remote site, linked-wrists, for our reference electrodes so that we could be certain that no significant volume conduction of potentials could occur from the source in the SCM to the reference site. When this was done we found that if the sternum was used as a reference site, as was the case in previous studies, some subjects exhibited bilateral responses while in others, the response was ipsilateral. However, with linked-wrists as the reference site, responses were always purely ipsilateral. Furthermore, recordings that used the sternum or the ipsilateral mastoid process as active sites and linked-wrists as a reference, exhibited responses which were inverted. Thus, both the sternum and the ipsilateral mastoid process are electrically active due to volume conduction from the nearby source in the SCM. The ambiguity in previous recordings can be attributed to the use of these active sites as a reference. When SCM responses are recorded versus a remote, electrically inactive site, the responses are purely ipsilateral.

In this study, we investigated the functions of PH-20 and acrosin during the interaction of macaque sperm with the zona pellucida. Both of these sperm enzymes have been reported to be present on the inner acrosomal membrane of acrosome reacted sperm, and have been suggested to play a role during secondary sperm-zona binding in other species. Anti-macaque PH-20 IgG, anti-pig acrosin IgG and soybean trypsin inhibitor (SBTI) were used as probes for immunolocalization of the two proteins at the ultrastructural level, and as reagents for blocking sperm penetration of the macaque zona pellucida in vitro. As a control, we performed similar studies with antibodies to CD-46, which is also located on the inner acrosomal membrane, but has no known function in sperm-zona pellucida interaction. After labeling with anti-acrosin IgG, gold label was not present on the sperm surface before the acrosome reaction, but was detected over the entire head of sperm that were induced to acrosome react with calcium ionophore A23187. In contrast, when sperm were induced to acrosome react by binding to intact zona pellucida, acrosin was present in the acrosomal shroud but not on the inner acrosomal membrane. Similar results were obtained when SBTI was used as a probe for enzyme localization. PH-20 and CD-46 were demonstrated on the inner acrosomal membrane of sperm induced to acrosome react by ionophore treatment and by zona binding. Neither anti-acrosin IgG nor anti-CD-46 IgG affected sperm penetration of the zona at concentrations up to 300 microg/ml, but zona penetration was blocked completely when anti-PH-20 IgG (100 microg/ml) was present during sperm-oocyte interaction. Ultrastructural observations of oocytes incubated with anti-PH-20 IgG showed that acrosomal shrouds were present on the zona surface but no sperm had begun to penetrate into the zona substance. We conclude that anti-PH-20 IgG prevented sperm penetration of the macaque zona pellucida by interference with secondary sperm-zona binding, rather than primary sperm-zona binding or the zona-induced acrosome reaction. Acrosin was not detected on the inner acrosomal membrane of sperm that are induced to acrosome react after zona binding, and acrosin does not appear to be critical for sperm penetration of the macaque zona pellucida.

Preparations of sperm membranes (plasma membranes and outer acrosomal membranes) and denuded sperm heads were isolated from macaque sperm, and the PH-20 proteins present were characterized by Western blotting, hyaluronic acid substrate gel analysis, and a microplate assay for hyaluronidase activity. Because we have shown previously that PH-20 is located on the plasma membrane and not on the outer acrosomal membrane, the PH-20 in the membrane preparations was presumed to be plasma membrane PH-20 (PM-PH-20). PM-PH-20 had an apparent molecular weight of 64 kDa and the optimum pH for its hyaluronidase activity was 6.5. The PH-20 associated with denuded sperm heads was localized by immunogold label to the persistent inner acrosomal membrane (IAM) and was presumed to be IAM-PH-20, which included a major 64 kDa form and a minor 53 kDa form. The 53 kDa form was not detected in extracts of denuded sperm heads from acrosome intact sperm that were boiled in nonreducing sample buffer, but was present in extracts of sperm heads from acrosome reacted sperm and in the soluble material released during the acrosome reaction, whether or not the samples were boiled. Substrate gel analysis showed that the hyaluronidase activity of the 53 kDa form of PH-20 was greatest at acid pH, and this activity was probably responsible for the broader and lower optimum pH of IAM hyaluronidase activity. When hypotonic treatment was used to disrupt the sperm acrosome and release the acrosomal contents, less than 0.05% of the total hyaluronidase activity was released. The PH-20 protein released by hypotonic treatment was the 64 kDa form and not the 53 kDa form, suggesting that its source might be the disrupted plasma membranes. Our experiments suggest that the soluble form of hyaluronidase, which is released at the time of the acrosome reaction, is derived from the IAM. This soluble hyaluronidase is composed of both the 64 kDa form and 53 kDa form of PH-20. The 53 kDa form appears to be processed from the 64 kDa form at the time of the acrosome reaction.

A microplate assay for hyaluronidase and a heterologous cumulus penetration assay were used to determine the effects of four flavonoids (tannic acid, kaempferol, quercetin, and apigenin) on the function of cynomolgus monkey sperm. All four flavonoids inhibited the activity of hyaluronidase extracted from monkey sperm in a concentration-dependent manner over the range of 50-200 microM. Tannic acid and apigenin had lower inhibitory effects than kaempferol and quercetin. Kaempferol, quercetin, and apigenin at 100 microM were shown to significantly inhibit monkey sperm penetration into hamster cumulus. There was a significant linear relationship between the capacity of the flavonoids to inhibit monkey sperm hyaluronidase activity and their inhibitory effects on hamster cumulus penetration (r = 0.97). Tannic acid was observed to reduce sperm motility, and it was not used in the cumulus penetration assay. The other three flavonoids tested in the cumulus penetration assay did not affect sperm motility, nor did they induce acrosome reactions. The results demonstrate that the flavonoids are useful tools for assessing the involvement of hyaluronidase in the functions of monkey sperm that are involved in fertilization.

PH-20 is a sperm plasma-membrane protein that has been shown to have hyaluronidase activity in several mammalian species including nonhuman primates. In this investigation, the PH-20 protein was characterized in noncapacitated human sperm and in capacitated human sperm. Two forms of PH-20 were observed in immunoblots of sodium dodecylsulfate polyacrylamide-gel electrophoresis (SDS PAGE) using a polyclonal antibody to recombinant PH-20: a major band of 64 kDa appeared in noncapacitated and capacitated sperm extracts and a 53-kDa band that appeared only in the acrosome-reaction supernatant of acrosome-reacted sperm. Using hyaluronic acid substrate gel analysis, we demonstrated that noncapacitated sperm extracts, capacitated sperm extracts, and the acrosome-reaction supernatant had hyaluronidase activity at neutral pH (pH 7) and acid pH (pH 4). The 64-kDa form in all samples had hyaluronidase activity at both neutral and acid pH, but the 53-kDa form was only active at acid pH. Total hyaluronidase activity, as measured by a microplate assay, was higher at pH 7 than at pH 4. Very low hyaluronidase activity was detected in the acrosome-reaction supernatant. Transmission electron microscopy and immunogold labeling showed that PH-20 of acrosome-intact human sperm was located on the plasma membrane over the entire head but not on the sperm midpiece and tail. After the acrosome reaction, PH-20 was also located on the inner acrosomal membrane. The biochemical characteristics and the ultrastructural localization of PH-20 in human sperm suggest that this protein is the human sperm hyaluronidase and, therefore, has an important function during fertilization.

Calcium was localized by in situ precipitation with potassium antimonate during epididymal maturation of the mouse and guinea pig sperms. In caput epididymis the calcium in mouse sperm head was mainly localized on the inner surface of tha outer acrosomal membrane (OAM) in preacrosomal region. During the passage from the caput epididymis to the cauda epididymis the calcium amount of mouse sperm did not undergo apparent changes. In comparison with mouse sperm, there were a few fine calcium deposite granules on the inner surface of the guinea pig sperm OAM on the abdomen side at the caput epididymis stage, but these granules disappeared at the cauda epididymis stage. The microvilli of columnar cell in columnar epithlium lining the epididymal duct are probably involved in regultion of Ca2+ concentration of the intraluminal fluid. The calcium precipitation granules distributing in the microvilli were observed. An abundance of Ca2+ were present in the intraluminal fluid of the corpus epididymis and they might have some important functions during the process of sperm maturation. Calcium in sperm tail was mainly distributed in the mitochondria. Compared with the mouse sperm, the mitochondria of guinea pig sperm possessed more calcium.

Background In vitro culture (IVC) and IVF of preimplantation mouse embryos are associated with changes in gene expression. It is however not known whether ICSI has additional effects on the transcriptome of mouse blastocysts. Methods We compared gene expression and development of mouse blastocysts produced by ICSI and cultured in Whitten's medium (ICSI(WM)) or KSOM medium with amino acids (ICSI(KSOMaa)) with control blastocysts flushed out of the uterus on post coital Day 3.5 (in vivo). In addition, we compared gene expression in embryos generated by IVF or ICSI using WM. Global pattern of gene expression was assessed using the Affymetrix 430 2.0 chip. Results Blastocysts from ICSI fertilization have a reduction in the number of trophoblastic and inner cell mass cells compared with embryos generated in vivo. Approximately 1000 genes are differentially expressed between ICSI blastocyst and in vivo blastocysts; proliferation, apoptosis and morphogenetic pathways are the most common pathways altered after IVC. Unexpectedly, expression of only 41 genes was significantly different between embryo cultured in suboptimal conditions (WM) or optimal conditions (KSOM(aa)). Conclusions Our results suggest that fertilization by ICSI may play a more important role in shaping the transcriptome of the developing mouse embryo than the culture media used.

210 cases of scrofulous sinus has been reported. Baluodan (BLD) has removed necrotic tissue and promote the granulation. It had potent inhibition on TB with no side effect in animal experiments and clinical trials. The total effective rate was 98.57%. The average of treatment course was 39.0 +/- 13.7 days. Follow-up for 2-4 years showed 3 cases had recurrence (1.4%). Its advantages were high effective rate, short curative course, low recurrence rate, with convenient in administration, and easy to spread.

Bridge failures in recent earthquakes such as the 1989 Loma Prieta earthquake have attracted the attention of the bridge engineering community to the large number of bridges with substandard seismic design details. Many concrete columns in bridges designed before the new seismic design provisions were adopted have low flexural ductility, low shear strength, and inadequate lap length for starter bars. These problems, compounded by flaws in the design of structural systems, have contributed to the catastrophic bridge failures in recent earthquakes. In this paper, a new technique for seismic strengthening of concrete columns is presented. The technique requires wrapping thin, flexible high-strength fiber composite straps around the column to improve the confinement and, thereby, its ductility and strength. Analytical models are presented that quantify the gain in strength and ductility of concrete columns externally confined by means of high-strength fiber composite straps. A parametric study is conducted to examine the effects of various design parameters such as concrete compressive strength, thickness and spacing of straps, and type of strap. The results indicate that the strength and ductility of concrete columns can be significantly increased by wrapping high-strength fiber composite straps around the columns.

The distribution of the soil fungi and the edaphic properties of the iron‐ore tailings along the eastern coast of Tolo Harbour, Hong Kong were studied. There was a phenomena of fungal succession according to the different ages of the tailing‐deposits. There was a gradual decline in the number of fungal species and the abundance of mycelia related to depth and the distance towards the seashore. The occurrence of the fungi was found to be governed by some of the edaphic properties tested, such as texture, content of organic carbon, pH, etc. The content of the metals seemed to play a minor role. In view of the lack of the basic nutrients in the tailings, it is suggested that sewage sludge be applied for stimulating the decomposition of organic matter and the subsequent release of nutrients will be available to higher plants.