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In this paper, we investigated how magnetic field influenced Trichel discharges by performing a statistical analysis of the current amplitude and pulse interval of Trichel discharges at various magnetic field intensities. Experimental results indicate that the amplitude and pulse intervals always obey lognormal functions. As the magnetic field intensity increased, the current amplitudes almost decreased and the pulse intervals increased significantly. The paths of negative ions produced in the Trichel discharge were curved by the magnetic field, weakening their shielding effect. A magnetic field can enhance the possibility of Trichel discharge with small energy release. Moreover, the characteristics of the Trichel pulse could be controlled with only magnetic field and without any energy addition.

The effect of the tube materials on the characteristics of an argon atmospheric pressure cold plasma jet is investigated through a single-electrode configuration driven by ac power supply. Two kinds of tube materials, Pyrex glass and 95% alumina ceramic, are applied in the experiments. Results show that, with the change of tube material, both the discharge current and the plume length are barely affected, whereas the plume volume exhibits an enormous distinction. In comparison with the results obtained in the ceramic tube, the peak volume can increase by 65% in the glass tube, while the increment of the peak length is only 2%. Through the analysis, the difference in the thermal conductivity between the glass and the ceramic is regarded as the main reason.

The stabilization characteristics of liftoff and reattachment in methane diffusion flames have been investigated experimentally with addition of plasmas. The flame liftoff and reattachment velocities have been measured by varying the applied voltages on plasmas. The results show that the liftoff and reattachment velocities are strongly affected by the plasmas. The difference between these two velocities as well as the flame hysteresis region is reduced and eventually disappeared with the addition of plasmas. A topological approach is put forward to explore the mathematic laws of hysteresis phenomena, and the existent hysteresis and catastrophe laws of flame structure transition are interpreted under the typical operation routes. It is concluded that a change of flame stabilization will occur when the fuel velocity or the applied voltage passes through stabilization boundaries in a special direction.

Atmospheric pressure discharges excited by repetitive nanosecond pulses have attracted significant attention for various applications. In this paper, a plate-plate discharge with airflows is excited by a repetitive nanosecond pulse generator. Under different experiment conditions, the applied voltages, discharge currents, and discharge images are recorded. The plasma images presented here indicate that the volume discharge modes vary with airflow speeds, and a diffuse and homogeneous volume discharge occurs at the speed of more than 35 m/s. The role of airflows provides different effects on the 2-stage pulse discharges. The 1st pulse currents nearly maintain consistency for different airflow speeds. However, the 2nd pulse current has a change trend of first decreasing and then rapidly increasing, and the value difference for 2nd pulse currents is about 20 A under different airflows. In addition, the experimental results are discussed according to the electrical parameters and discharge images.

Autosomal dominant polycystic kidney disease pathogenesis can be recapitulated in animal models by gene mutations in or dosage alterations of polycystic kidney disease 1 ( PKD1 ) or PKD2 , demonstrating that too much and too little PKD1/PKD2 are both pathogenic. Gene dosage manipulation has become an appealing approach by which to compensate for loss or gain of gene function, but the mechanisms controlling PKD2 expression remain incompletely characterized. In this study, using cultured mammalian cells and dual-luciferase assays, we found that the 3′ untranslated region (3′UTR) of PKD2 mRNA inhibits luciferase protein expression. We then identified nucleotides 691–1044, which we called 3FI, as the 3′UTR fragment necessary for repressing the expression of luciferase or PKD2 in this system. Using a pull-down assay and mass spectrometry we identified far upstream element-binding protein 1 (FUBP1) as a 3FI-binding protein. In vitro overexpression of FUBP1 inhibited the expression of PKD2 protein but not mRNA. In embryonic zebrafish, FUBP1 knockdown (KD) by morpholino injection increased PKD2 expression and alleviated fish tail curling caused by morpholino-mediated KD of PKD2. Conversely, FUBP1 overexpression by mRNA injection significantly increased pronephric cyst occurrence and tail curling in zebrafish embryos. Furthermore, FUBP1 binds directly to eukaryotic translation initiation factor 4E-binding protein 1, indicating a link to the translation initiation complex. These results show that FUBP1 binds 3FI in the PKD2 3′UTR to inhibit PKD2 translation, regulating zebrafish disease phenotypes associated with PKD2 KD.

Summary Transient receptor potential vanilloid 6 (TRPV6), a calcium-selective channel possessing six transmembrane domains (S1-S6) and intracellular N and C termini, plays crucial roles in calcium absorption in epithelia and bone and is involved in human diseases including vitamin-D deficiency, osteoporosis, and cancer. The TRPV6 function and regulation remain poorly understood. Here we show that the TRPV6 intramolecular S4-S5 linker to C-terminal TRP helix (L/C) and N-terminal pre-S1 helix to TRP helix (N/C) interactions, mediated by Arg470:Trp593 and Trp321:Ile597 bonding, respectively, are autoinhibitory and are required for maintaining TRPV6 at basal states. Disruption of either interaction by mutations or blocking peptides activates TRPV6. The N/C interaction depends on the L/C interaction but not reversely. Three cationic residues in S5 or C terminus are involved in binding PIP2 to suppress both interactions thereby activating TRPV6. This study reveals “PIP2 - intramolecular interactions” regulatory mechanism of TRPV6 activation-autoinhibition, which will help elucidating the corresponding mechanisms in other TRP channels., Graphical Abstract, Highlights • The R470:W593 and W321:I597 residue pairs mediate the L/C and N/C interactions in TRPV6 • The L/C and N/C interactions are inhibitory for the TRPV6 function • PIP2 binds to cationic residues in the S5 helix and C terminus of TRPV6 • PIP2 activates TRPV6 by disrupting the L/C and N/C interactions, Biochemistry; Neuroscience

Scramjet is the optimal propulsion system for hypersonic vehicles, and it is important to study its maximum flight Mach number. Upper limit and its influence factors of the flight Mach number of dual-mode scramjet used in hypersonic vehicles are investigated in this paper. Scramjet performance analysis model with the constant-area heating process, cooling requirement analysis model of the combustor and cruise model of the scramjet-powered hypersonic vehicle are established. Specific impulse of dual-mode scramjet engine with hydrocarbon fuel is similar to that of liquid oxygen kerosene rocket engine when the freestream Mach number is above 10. The ratio of hypersonic vehicle reference area to scramjet inlet capture area is introduced to establish the size relationship between the scramjet engine and the hypersonic vehicle. Considering the balance of thrust and drag, smaller ratio of hypersonic vehicle reference area to scramjet inlet capture area leads to higher flight Mach number that hypersonic vehicle can cruise. When the angle of attack is 10 degrees, the ratio of hypersonic vehicle reference area to scramjet capture area should be less than 5.5 to ensure that the vehicle can work at Mach 8. The maximum length to diameter ratio of the combustor is used to indicate active cooling requirements, and it is less than 5 under the cooling requirement when flight Mach number is above 9. Reducing the heat flux at the combustor wall and increasing the fuel heat sink is helpful to increase the allowable combustion chamber length, so as to increase the maximum flight Mach number of the scramjet engine on the premise of meeting thermal protection requirement.

PKD2 and PKD1 genes are mutated in human autosomal dominant polycystic kidney disease. PKD2 can form either a homomeric cation channel or a heteromeric complex with the PKD1 receptor, presumed to respond to ligand(s) and/or mechanical stimuli. Here, we identify a two-residue hydrophobic gate in PKD2L1, and a single-residue hydrophobic gate in PKD2. We find that a PKD2 gain-of-function gate mutant effectively rescues PKD2 knockdown-induced phenotypes in embryonic zebrafish. The structure of a PKD2 activating mutant F604P by cryo-electron microscopy reveals a π- to α-helix transition within the pore-lining helix S6 that leads to repositioning of the gate residue and channel activation. Overall the results identify hydrophobic gates and a gating mechanism of PKD2 and PKD2L1., Mutations in the cation channel PKD2 cause human autosomal dominant polycystic kidney disease but its channel function and gating mechanism are poorly understood. Here authors study PKD2 using electrophysiology and cryo-EM, which identifies hydrophobic gates and proposes a gating mechanism for PKD2.

An experimental investigation of combustion mode transition in a hydrocarbon fueled scramjet combustor model is reported under Mach number 2.1 and 2.5 inflow conditions. Three different combustion modes with respect to equivalence ratio are observed, namely, scramjet mode, weak ramjet mode and strong ramjet mode. The typical features of different combustion modes are analyzed by wall-pressures and one-dimensionally estimated Mach number distributions. The processes of combustion mode transitions show significant nonlinear characteristics. The static pressure and Mach number have discontinuous sudden changes as the mode transition occurs, especially near the fuel-supply region, emphasizing the importance of detection and control of combustion modes. The nonlinear characteristics of wall-pressures near the exit of the isolator can be used in the detection of different combustion modes. A series of experiments prove that this pressure-magnitude-based detection technique is feasible.

Scramjet engine encounters many safety restrictions during the ascent trajectory. These restrictions make the engine a multi-objective control system. In order to obtain optimal thrust performance while avoiding the inlet unstart, switching control based on Min strategy has been focused on in the working process of scramjet engine. Firstly, on the basis of experimental data, the dynamic mathematical models of fuel supply system, thrust regulation control loop and inlet unstart protection loop were established. Min strategy and PI controller were applied to the scramjet engine and the influence of integral limiter of PI controllers on the control process was discussed using the hardware-in-the-loop simulation. Finally, the validity of switching control was verified on the ground experiment facility. The results showed that it is useful to introduce the multi-objective switching control to scramjet engine. Switching control based on Min strategy could effectively maximize the engine performance while avoiding the inlet unstart.

Treponema pallidum, hepatitis C virus (HCV), human immunodeficiency virus (HIV)-1, and hepatitis B virus (HBV) are major causes of sexually transmitted diseases passed through blood contact. The development of a sensitive and efficient method for detection is critical for early diagnosis and for large-scale screening of blood specimens in China. This study aims to establish an assay to detect these pathogens in clinical serum specimens. We established a TaqMan-locked nucleic acid (LNA) real-time polymerase chain reaction (PCR) assay for rapid, sensitive, specific, quantitative, and simultaneous detection and identification. The copy numbers of standards of these 4 pathogens were quantified. Standard curves were generated by determining the mean cycle threshold values versus 10-fold serial dilutions of standards over a range of 10(6) to 10(1) copies/μL, with the lowest detection limit of the assay being 10(1) copies/μL. The assay was applied to 328 clinical specimens and compared with enzyme-linked immunosorbent assay (ELISA) and commercial nucleic acid testing (NAT) methods. The assay identified 39 T. pallidum-, 96 HCV-, 13 HIV-1-, 123 HBV-, 5 HBV/HCV-, 1 T. pallidum/HBV-, 1 HIV-1/HCV-, and 1 HIV-1/T. pallidum-positive specimens. The high sensitivity of the assay confers strong potential for its use as a highly reliable, cost-effective, and useful molecular diagnostic tool for large-scale screening of clinical specimens. This assay will assist in the study of the pathogenesis and epidemiology of sexually transmitted blood diseases.

Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in PKD1 or PKD2 which encodes polycystin-1 (PC1) and polycystin-2, respectively. PC1 was previously shown to slow cell proliferation and inhibit apoptosis but the underlying mechanisms remain elusive or controversial. Here we showed in cultured mammalian cells and Pkd1 knockout mouse kidney epithelial cells that PC1 and its truncation mutant comprising the last five transmembrane segments and the intracellular C-terminus (PC1-5TMC) down-regulate the phosphorylation of protein kinase R (PKR) and its substrate eukaryotic translation initiation factor 2 alpha (eIF2α). PKR is known to be activated by interferons and dsRNAs, inhibits protein synthesis and induces apoptosis. By co-immunoprecipitation experiments we found that PC1 truncation mutants associate with PKR, or with PKR and its activator PACT. Further experiments showed that PC1 and PC1-5TMC reduce phosphorylation of eIF2α through inhibiting PKR phosphorylation. Our TUNEL experiments using tunicamycin, an apoptosis inducer, and GADD34, an inhibitor of eIF2α phosphorylation, demonstrated that PC1-5TMC inhibits apoptosis of HEK293T cells in a PKR-eIF2α-dependent manner, with concurrent up- and down-regulation of Bcl-2 and Bax, respectively, revealed by Western blotting. Involvement of PC1-regulated eIF2α phosphorylation and a PKR-eIF2α pathway in cell apoptosis may be an important part of the mechanism underlying ADPKD pathogenesis.

// Cefan Zhou 1, 2 , Jing Yu 2, 3 , Ming Wang 4 , Jing Yang 5 , Hui Xiong 6 , Huang Huang 1 , Dongli Wu 2 , Shimeng Hu 2 , Yefu Wang 2 , Xing-Zhen Chen 1, 7 and Jingfeng Tang 1 1 Institute of Biomedical and Pharmaceutical Sciences, Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Hubei Key Laboratory of Industrial Microbiology, Hubei University of Technology, Wuhan, Hubei, China 2 The State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei, China 3 Department of Clinical Laboratory, Hubei Cancer Hospital, Wuhan, Hubei, China 4 Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, Hubei, China 5 Institute for Immunology, Tsinghua University, Beijing, China 6 XiLi People's Hospital, Shenzhen, Guangdong, China 7 Membrane Protein Disease Research Group, Department of Physiology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada Correspondence to: Jingfeng Tang, email: Jingfeng_HUT@163.com Keywords: prognostic significance, biomarker, cell proliferation, survival, meta-analysis Received: July 05, 2017 Accepted: August 27, 2017 Published: September 19, 2017 ABSTRACT In the present study, we found the mRNA expression level of glycerol-3-phosphate dehydrogenase (GPD1) was significantly downregulated in human breast cancer patients. Patients with reduced GPD1 expression exhibited poorer overall metastatic relapse-free survival ( p = 0.0013). Further Cox proportional hazard model analysis revealed that the reduced expression of GPD1 is an independent predictor of overall survival in oestrogen receptor-positive ( p = 0.0027, HR = 0.91, 95% CI = 0.85–0.97, N = 3,917) and nodal-negative ( p = 0.0013, HR = 0.87, 95% CI = 0.80–0.95, N = 2,456) breast cancer patients. We also demonstrated that GPD1 was a direct target of miR-370, which was significantly upregulated in human breast cancer. We further showed that exogenous expression of GPD1 in human MCF-7 and MDA-MB-231 breast cancer cells significantly inhibited cell proliferation, migration, and invasion. Our results, therefore, suggest a novel tumour suppressor function for GPD1 and contribute to the understanding of cancer metabolism.

Herpes simplex virus type 2 (HSV-2) is associated with a variety of diseases that are health problems worldwide. Our early study showed that lambda-interferons (IFN-λs), induced by the activation of the Toll-like receptor 3 and retinoic acid-inducible protein I signaling pathways, contribute to inhibition of HSV-2 replication in human cervical epithelial cells. However, anti-HSV-2 mechanisms and specific differences in signaling transduction by different IFN-λs in human cervical epithelial cells remain unclear. In this study, we demonstrated potent inhibition of HSV-2 replication by IFN-λs without cytotoxicity. Investigation of the underlying mechanism(s) showed that IFN-λs induced expression of IFN-stimulated genes (ISGs) and enhanced the expression of several pattern recognition receptors (PRRs). Among the IFN-λs, IFN-λ3 induced higher levels of ISG and PRR expression. In addition, IFN-λs up-regulated a number of genes that encode components of the Janus kinase signal transducers and activators of transcription (JAK/STAT) signaling pathway. Inhibition of the JAK/STAT signaling pathway by a JAK inhibitor abolished IFN-λ-mediated anti-HSV-2 activity and induction of ISGs and PRRs, whereas the induction of ISGs and PRRs by IFN-λs was not compromised by HSV-2 infection. These findings provide further experimental evidence that IFN-λs have therapeutic potential for HSV-2 infections.

A MHD generator with different arrangements of electromagnetic fields will lead the generator working in three modes. A quasi-one-dimensional approximation is used for the model of the MHD generator to analyze the inner mechanism of operation modes. For the MHD generator with a uniform constant magnetic field, a specific critical electric field Ecr is required to decelerate a supersonic entrance flow into a subsonic exit flow. Otherwise, the generator works in a steady mode with a larger electric field than Ecr in which a steady supersonic flow is provided at the exit, or the generator works in a choked mode with a smaller electric field than Ecr in which the supersonic entrance flow is choked in the channel. The detailed flow field characteristics in different operation modes are discussed, demonstrating the relationship of operation modes with electromagnetic fields.

PKD2 and PKD1 genes are mutated in human autosomal dominant polycystic kidney disease. PKD2 can form either a homomeric cation channel or a heteromeric complex with the PKD1 receptor, presumed to respond to ligand(s) and/or mechanical stimuli. Here, we identify a two-residue hydrophobic gate in PKD2L1, and a single-residue hydrophobic gate in PKD2. We find that a PKD2 gain-of-function gate mutant effectively rescues PKD2 knockdown-induced phenotypes in embryonic zebrafish. The structure of a PKD2 activating mutant F604P by cryo-electron microscopy reveals a π- to α-helix transition within the pore-lining helix S6 that leads to repositioning of the gate residue and channel activation. Overall the results identify hydrophobic gates and a gating mechanism of PKD2 and PKD2L1.

Surrounded with distilled water, a classic argon atmospheric pressure discharge device is employed to study the propagation of the plasma. By examining the high-speed photographs, a phenomenon of interruption is found at different flow rates. At 3 L/min, the interruption appears only at the downward stage of the applied voltage after the plasma jet has run into the ambient air. As the flow rate reaches up to 15 L/min, however, the interruption can be obtained at a rather low applied voltage and exists in both the upward and downward stages. It is believed that the active species flowing from the upstream to the downstream region may be the main cause.

// Cefan Zhou 1, 2, * , Hongxia Cheng 3, * , Wenying Qin 1, * , Yi Zhang 1 , Hui Xiong 4 , Jing Yang 5 , Huang Huang 1 , Yefu Wang 2 , Xing-Zhen Chen 1, 6 , Jingfeng Tang 1 1 Institute of Biomedical and Pharmaceutical Sciences, Key Laboratory of Fermentation Engineering (Ministry of Education), College of Bioengineering, Hubei University of Technology, Wuhan, 430068, China 2 The State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, China 3 Department of Chemical and Pharmaceutical Engineering, Wuhan Huaxia University of Technology, Wuhan, 430223, China 4 XiLi People's Hospital, Shenzhen, Guangdong, 518055, China 5 Institute for Immunology, Tsinghua University, Beijing, 100084, China 6 Membrane Protein Disease Research Group, Department of Physiology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, T6G 2R3, Canada * These authors contributed equally to this work Correspondence to: Jingfeng Tang, email: Jingfeng_HUT@163.com Keywords: Pygo2, paclitaxel, MDR1, P-glycoprotein, glioma Received: November 16, 2016 Accepted: February 20, 2017 Published: March 02, 2017 ABSTRACT Anti-microtubule drugs, such as paclitaxel (PTX), are extensively used for the treatment of numerous cancers. However, growing evidence has shown that PTX resistance, either intrinsic or acquired, frequently occurs in patients and results in the failure of treatment, contributing to the high cancer mortality rate. Therefore, it is necessary to identify the genes or pathways involved in anti-microtubule drug resistance for future successful treatment of cancers. Pygopus2 (Pygo2), which contains a Zn-coordinated plant homeodomain (PHD) finger domain, is critical for β-catenin-dependent transcriptional switches in normal and malignant tissues and is over-expressed in various cancers, including human brain glioma. In this study, we report that over-expression of Pygo2 inhibited the efficacy of PTX and contributed to cell multidrug resistance in two different ways. First, over-expression of Pygo2 inhibited the PTX-induced phosphorylation of B-cell lymphoma 2 (Bcl-2), suppressing the proteolytic cleavage of procaspase-8/9 and further inhibiting the activation of caspase-3, which also inhibits the activation of the JNK/SAPK pathway, ultimately inhibiting cell apoptosis. Second, over-expression of Pygo2 facilitated the expression of P-glycoprotein, which acts as a drug efflux pump, by promoting the transcription of Multi-drug resistance 1 (MDR1) at the MDR1 promoter loci, resulting in acceleration of the efflux of PTX.

Transient receptor potential polycystin-3 (TRPP3) is a cation channel activated by calcium and proton and is involved in hedgehog signaling, intestinal development, and sour tasting. How TRPP3 channel function is regulated remains poorly understood. By N-terminal truncation mutations, electrophysiology, and Xenopus oocyte expression, we first identified fragment Asp-21-Ser-42 to be functionally important. We then found that deletion mutant Δ1-36 (TRPP3 missing fragment Met-1-Arg-36) has a similar function as wild-type TRPP3, whereas Δ1-38 is functionally dead, suggesting the importance of Val-37 or Cys-38. Further studies found that Cys-38, but not Val-37, is functionally critical. Cys-38 is a predicted site of palmitoylation, and indeed TRPP3 channel activity was inhibited by palmitoylation inhibitor 2-bromopalmitate and rescued by palmitoylation substrate palmitic acid. The TRPP3 N terminus (TRPP3NT, Met-1-Leu-95) localized along the plasma membrane of HEK293 cells but stayed in the cytoplasm with 2-bromopalmitate treatment or C38A mutation, indicating that TRPP3NT anchors to the surface membrane through palmitoylation at Cys-38. By acyl-biotin exchange assays, we showed that TRPP3, but not mutant C38A, is indeed palmitoylated. When putative phosphorylation sites near Cys-38 were mutated to Asp or Glu to mimic phosphorylation, only T39D and T39E reduced TRPP3 function. Furthermore, TRPP3NT displayed double bands in which the upper band was abolished by λ phosphatase treatment or T39A mutation. However, palmitoylation at Cys-38 and phosphorylation at Thr-39 independently regulated TRPP3 channel function, in contrast to previous reports about correlated palmitoylation with a proximate phosphorylation. Palmitoylation at Cys-38 represents a novel mechanism of functional regulation for TRPP3.

Breast cancer is a heterogeneous disease associated with diverse clinical, biological and molecular features, presenting huge challenges for prognosis and treatment. Here we found that perilipin-1 (PLIN1) mRNA expression is significantly downregulated in human breast cancer. Kaplan-Meier analysis indicated that patients presenting with reduced PLIN1 expression exhibited poorer overall metastatic relapse-free survival (p = 0.03). Further Cox proportional hazard models analysis revealed that the reduced expression of PLIN1 is an independent predictor of overall survival in estrogen receptor positive (p < 0.0001, HR = 0.87, 95% CI = 0.81-0.92, N = 3,600) and luminal A-subtype (p = 0.02, HR = 0.88, 95% CI = 0.78-0.98, N = 1,469) breast cancer patients. We also demonstrated that the exogenous expression of PLIN1 in human breast cancer MCF-7 and MDA-MB-231 cells significantly inhibits cell proliferation, migration, invasion and in vivo tumorigenesis in mice. Together, these data provide novel insights into a prognostic significance of PLIN1 in human breast cancer and reveal a potentially new gene therapy target for breast cancer.

Infection with human papillomavirus (HPV), particularly HPV16 and HPV18, is the main cause of invasive cervical cancer, although other factors such as herpes simplex virus (HSV) may act in conjunction with HPV in this context. To explore the possibility of developing a system for rapid diagnosis and clinical screening of cervical cancer, we developed a multiplex real-time PCR assay that can simultaneously detect and quantify HPV16/18 and HSV1/2. To evaluate its possibilities and practical uses, 177 samples collected from patients with suspected HPV and HSV infection in exfoliated cervical cells, genital herpes or labial herpes were tested by multiplex real-time PCR and compared with results obtained by DNA sequencing. Each virus was detected over a range from 1.0 × 10 1 to 1.0 × 10 7 copies/reaction. The clinical sensitivity was 100% for HPV16/18 and HSV1/2. The clinical specificity was 97.1% for HPV16, 98.1% for HPV18, 97.0% for HSV1 and 96.0% for HSV2. The kappa value was 0.96 for HPV16, 0.92 for HPV18, 0.94 for HSV1 and 0.93 for HSV2, when DNA sequencing was used as the reference standard. In summary, this novel multiplex real-time PCR allows the rapid and specific detection of HPV16/18 and HSV1/2, as well as coinfection with HPV and HSV, in clinical samples. In the future, this multiplex real-time PCR assay will assist in cervical cancer screening, viral treatment evaluation and epidemiological studies in which high throughput analysis is required.

Kocuria flava HO-9041 isolated from the air of Xinjiang of China, is a Gram-positive, aerobic, coccoid, nonencapsulated, non-halophilic, and non-endospore-forming, in the genus Kocuria, and it shows the ability to remove heavy metal. Here, we report the finished and annotated genome sequence of this organism.

To explore the possibilities of a novel multiplex real-time PCR system for rapid diagnosis, genetic typing of serovars and clinical application in NGU, we developed a multiplex real-time PCR system for the simultaneous diagnosis of Chlamydia trachomatis, Ureaplasma parvum and Ureaplasma urealyticum and molecular detection of serovars of C. trachomatis and U. parvum in NGU using the SNP technology and TaqMan-LNA probe. In 57 pathogen-positive clinical specimens, we identified the following C. trachomatis serovars: D (20.05%, 12/57), E (36.84%, 21/57), F (19.30%, 11/57), G (8.77%, 5/57), H (5.26%, 3/57), J (3.51%, 2/57), and K (5.26%, 3/57). In 115 pathogen-positive clinical specimens, we identified the following U. parvum serovars: 1 (0.87%, 2/115), 3 (55.65%, 64/115), 6 (20.87%, 24/115) and 14 (21.74%, 25/115). Our fast pathogen diagnosis and serotyping assay using real-time TaqMan-LNA PCR may improve our ability to study the pathogenesis and epidemiology of NGU.

This paper illustrates a repetitive nanosecond volume discharge in the airflows at different velocities. The plasma images presented here show that the volume discharge modes vary with the changes of airflow speeds, and a diffuse and homogeneous volume discharge occurs at the airflow speed of 30 m/s. The discharge is promoted in the airflows at a lower velocity, and the discharge tends to be inhibited at a speed to 40 m/s. However, the inhibited mode disappears at higher repetitive frequency of the pulse.

Based on gold-labeled silver stain (GLSS) method, we developed the visual protein microarray for simultaneous, sensitive, and specific detection of Ureaplasma parvum and Chlamydia trachomatis using N-terminus multiple-banded antigen (NMBA) of U. parvum and major outer membrane protein of C. trachomatis. The specific antigens were immobilized on glass surface that was treated with 3-glycidoxypropyltrimethoxysilane, and they were used as the capturing probes to recognize the complementary target antibodies binding to the detecting probes of Nano-gold-Staphylococcal protein A (SPA). In the "sandwich" format, Nano-gold-SPA probe was used as an indicator and GLSS was applied to amplify the detection signals and produce black image on array spots, which were visible with naked eyes. In our model arrays, the detection limit of protein microarray was as low as 2 ng/mL, and the lowest titer of detectable antibody was 1:128; thus, this sensitivity was comparable to the fluorescent detection method. The visual simultaneous protein microarrays were used to detect total 186 clinical samples, which had been determined by enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative real-time polymerase chain reaction; the results were identical and no distinct difference (P > 0.05) existed between them. Our results demonstrate that we have developed the visual protein microarray technique, which is of high sensitivity and high specificity, and it may have potential in clinical applications.

Based on gold label silver stain and coupled with multiplex asymmetric polymerase chain reaction (PCR) analysis, we developed the visual DNA microarray for simultaneous, sensitive, and specific detection of human immunodeficiency virus type-1 (HIV-1) and Treponema pallidum. The 5'-end amino-modified oligonucleotides were immobilized on glass surface, which were used as the capturing probes to bind the complement biotinylated target DNA. The gold-conjugated streptavidins were introduced to the microarray for specific binding to biotin. The black image of microarray spots, which were the result from the precipitation of silver onto nanogold particles and bound to streptavidins, was visualized and accounted as the detection of biotinylated target DNA. Multiplex asymmetric PCR products of HIV-1 and T. pallidum and Bacillus subtilis (used as positive control) were performed for preparing the abundant biotinylated single-stranded target DNA of which could affect detection efficiency and sensitivity of hybridization on microarray. One hundred sixty-nine clinical samples of HIV-1 and T. pallidum from infected patients were tested using the homemade DNA microarrays. The results were identical to those shown in the assays of ELISA and fluorescence quantitative real-time PCR. Our results demonstrate that we have developed the visual gene detection technique, which is of high sensitivity and specificity; it may have potential in clinical applications.

Berberine, a hypoglycemic agent, has recently been shown to activate AMP-activated protein kinase (AMPK) contributing to its beneficial metabolic effects in peripheral tissues. However, whether berberine exerts a regulatory effect on β-cells via AMPK or other signaling pathways and counteracts glucolipotoxicity remains uncertain. In the present study, the impact of berberine on β-cell function was investigated in vivo and in vitro. In high-fat-fed rats, berberine treatment for 6 wk significantly decreased plasma glucose and insulin levels before and after an oral glucose challenge along with the reduction of body weight and improvement of blood lipid profile. In accordance with the in vivo results, berberine acutely decreased glucose-stimulated insulin secretion (GSIS) and palmitate-potentiated insulin secretion in MIN6 cells and rat islets. However, pretreated with berberine for 24 h augmented the response of MIN6 cells and rat islets to glucose and attenuated the glucolipotoxicity. Berberine acutely increased AMPK activity in MIN6 cells. However, compound C, an AMPK inhibitor, completely reversed troglitazone-suppressed GSIS, not berberine-suppressed GSIS. Otherwise, berberine decreased cAMP-raising agent-potentiated insulin secretion in MIN6 cells and rat islets. These results suggest that the activation of AMPK is required for troglitazone-suppressed GSIS, whereas cAMP signaling pathway contributes, at least in part, to the regulatory effect of berberine on insulin secretion.

Polycystic kidney disease (PKD) protein 2 Like 1 (PKD2L1), also called transient receptor potential polycystin-3 (TRPP3), regulates Ca2+-dependent hedgehog signalling in primary cilia, intestinal development and sour tasting but with an unclear mechanism. PKD2L1 is a Ca2+-permeable cation channel that is activated by extracellular Ca2+ (on-response) in Xenopus oocytes. PKD2L1 co-expressed with PKD protein 1 Like 3 (PKD1L3) exhibits extracellular acid-induced activation (off-response, i.e., activation following acid removal) but whether PKD1L3 participates in acid sensing remains unclear. Here we used the two-microelectrode voltage-clamp, site directed mutagenesis, Western blotting, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunofluorescence and showed that PKD2L1 expressed in oocytes exhibits sustained off-response currents in the absence of PKD1L3. PKD1L3 co-expression augmented the PKD2L1 plasma membrane localization but did not alter the observed properties of the off-response. PKD2L1 off-response was inhibited by an increase in intracellular Ca2+. We also identified two intra-membrane residues aspartic acid 349 (D349) and glutamic acid 356 (E356) in the third transmembrane domain that are critical for PKD2L1 channel function. Our study suggests that PKD2L1 may itself sense acids and defines off-response properties in the absence of PKD1L3.

Pygo2 has been discovered as an important Wnt signaling component contributing to the activation of Wnt-target gene transcription. In the present study, we discovered that Pygo2 mRNA and protein levels were up-regulated in the majority of (152/209) human brain glioma tissues and five glioma cell lines and significantly correlated with the age, the WHO tumor classification and poor patient survival. The histone methyltransferase complex components (WDR5, Ash2 and menin, but not CXCC1 or NCOA6) were down-regulated at the promoter loci of Wnt target genes after Pygo2 knockdown and this was accompanied by the down-regulation of Wnt/β-catenin pathway activity. Further, we demonstrated that the involvement of Pygo2 in the activation of the Wnt pathway in human glioma progression is through up-regulation of the H3K4me3 (but not H3K4me2) by promoting the recruitment of the histone methyltransferase MLL1/MLL2 complex to Wnt target gene promoters. Thus, our study provided evidence that Pygo2 functions as a novel prognostic marker and represents a potential therapeutic target.

Lots of research have been carried out since Trichel pulse was presented in 19341,2. In order to obtain some stochastic properties of Trichel pulse in wire-plate negative corona discharge, statistical method is used in this paper to analyze three characteristic parameters of Trichel pulse, which are amplitude A, rising time t r , half-falling time t f .

In this work, effect of dielectric temperature on atmospheric pressure nonequilibrium plasma jets (APNPJs) with different dielectric materials has been investigated1–2. In our experiments, a single-electrode plasma generation equipped with an AC power supply is applied. Two materials, 95% alumina cerami c and quartz are adopted as dielectric tube. To distinguish the APNPJ status from the argon flow rate, the three modes, laminar, transition and turbulent, are separated.

As an emerging source, atmospheric pressure nonthermal plasma jets (APNPJs) have held remarkable promise owing to its wide applications1. At low pressure (∼0.1 Pa), magnetic field i s always introduced into plasmas to affect the discharge current2. However, little works have been reported on the interaction between magnetic field and atmospheric pressure plasmas. In this work, effect of magnetic field on the propagation of Ar APNPJs is investigated through a single-electrode configuration by use of AC power. In order to avoid the interference of the electric field by the permanent magnet, a same size iron is set i n the electric field to replace the permanent magnet when there's no magnetic field. We compare the electrical characteristic with/without iron and with/without magnetic field. As seen in Fig. 1, with the magnetic field, the discharge current is clearly increased, however, with and without the iron, the discharge current show slight difference. It is believed that the enhancement of ionization in the curved and lengthened electron path caused by Lorentz force is the main reason.

As a transient receptor potential (TRP) superfamily member, polycystic kidney disease 2-like-1 (PKD2L1) is also called TRPP3 and has similar membrane topology as voltage-gated cation channels. PKD2L1 is involved in hedgehog signaling, intestinal development and sour tasting. PKD2L1 and PKD1L3 form heterotetramers with 3:1 stoichiometry. C-terminal coiled-coil-2 (CC2) domain (G699-W743) of PKD2L1 was reported to be important for its trimerization but independent studies showed that CC2 does not affect PKD2L1 channel function. It thus remains unclear how PKD2L1 proteins oligomerize into a functional channel. By SDS-PAGE, blue native PAGE and mutagenesis we here identified a novel C-terminal domain called C1 (K575-T622) involved in stronger homotrimerization than the non-overlapping CC2 and found that the PKD2L1 N-terminus is critical for dimerization. By electrophysiology and Xenopus oocyte expression, we found that C1, but not CC2, is critical for PKD2L1 channel function. Our co-immunoprecipitation and dynamic light scattering experiments further supported involvement of C1 in trimerization. Further, C1 acted as a blocking peptide that inhibits PKD2L1 trimerization as well as PKD2L1 and PKD2L1/PKD1L3 channel function. Thus, our study identified C1 as the first PKD2L1 domain essential for both PKD2L1 trimerization and channel function and suggest that PKD2L1 and PKD2L1/PKD1L3 channels share the PKD2L1 trimerization process.

Polycystin-2 (PC2), encoded by the PKD2 gene, is mutated in ~15% of autosomal dominant polycystic kidney disease. Filamins are actin-binding proteins implicated in scaffolding and membrane stabilization. Here we studied the effects of filamin on PC2 stability using filamin-deficient human melanoma M2, filamin-A (FLNA)-replete A7, HEK293 and IMCD cells together with FLNA siRNA/shRNA knockdown (KD). We found that the presence of FLNA is associated with higher total and plasma membrane PC2 protein expression. Western blotting analysis in combination with FLNA KD showed that FLNA in A7 cells represses PC2 degradation, prolonging the half-life from 2.3 to 4.4 hours. By co-immunoprecipitation and Far Western blotting we found that the FLNA C-terminus (FLNAC) reduces the FLNA-PC2 binding and PC2 expression, presumably through competing with FLNA for binding PC2. We further found that FLNA mediates PC2 binding with actin through forming complex PC2-FLNA-actin. FLNAC acted as a blocking peptide and disrupted the link of PC2 with actin through disrupting the PC2-FLNA-actin complex. Finally, we demonstrated that the physical interaction of PC2-FLNA is Ca-dependent. Taken together, our current study indicates that FLNA anchors PC2 to the actin cytoskeleton through complex PC2-FLNA-actin to reduce degradation and increase stability, and possibly regulate PC2 function in a Ca-dependent manner.

Aim: To investigate the expression of feeding-related peptide receptors mRNA in GT1-7 cell line and roles of leptin and orexins in the control of GnRH secretion. Methods: Receptors ofbombesin3, cholecystokinin (CCK)-A, CCK-B, glucagon-like peptide (GLP)1, melanin-concentrating hormone (MCH)1, orexin1, orexin2, neuromedin-B, neuropeptide Y (NPY) 1 and NPY5, neurotensin (NT) 1, NT2, NT3, and leptin receptor long form mRNA in GT1-7 cells were detected by reversed transcriptase-polymerase chain reaction. GT1-7 cells were treated with leptin, orexin A and orexin B at a cohort of concentrations for different lengths of time, and GnRH in medium was determined by radioimmunoassay (RIA). Results: Receptors of bombesin 3, CCK-B, GLP1, MCH1, orexin1, neuromedin-B, NPY1, NPY5, NT1, NT3, and leptin receptor long form mRNA were expressed in GT1-7 cells, of which, receptors of GLP1, neuromedin-B, NPY1, and NT3 were highly expressed. No amplified fragments of orexin2, NT2, and CCK-A receptor cDNA were generated with GT1-7 RNA, indicating that the GT1-7 cells did not express mRNA of them. Leptin induced a significant stimulation of GnRH release, the results being most significant at 0.1 nmol/L for 15 min. In contrast to other studies in hypothalamic explants, neither orexin A nor orexin B affected basal GnRH secretion over a wide range of concentrations ranging from 1 nmol/L to 500 nmol/Lat 15, 30, and 60 min. Conclusion: Feeding and reproductive function are closely linked. Many orexigenic and anorexigenic signals may control feeding behavior as well as alter GnRH secretion through their receptors on GnRH neurons.

Rufibacter tibetensis strain 1351, isolated from the soil of the Tibet plateau of China, belongs to the family of Cytophagaceae. It is a red-pigmented, gram-negative, strictly aerobic and rod-shaped bacterium and shows resistance to UV radiation. Here, we report its complete genome sequence, which can help us find the key genes of the carotenoid biosynthesis and resistance to UV radiation.

Mutations in polycystin-1, polycystin-2, or fibrocystin account for autosomal dominant or recessive polycystic kidney disease. Renal cystogenesis is linked to abnormal localization and function of these cystoproteins in renal primary cilia. They are also expressed in extrarenal tissues in which their functions are unclear. Here we found that human type-II alveolar epithelial A549, airway submucosal Calu-3 cells, and rat bronchioles contain primary or multiple cilia in which we detected these cystoproteins. At sub-confluency, polycystin-1 was expressed on plasma membrane, while polycystin-2 was localized to the ER of resting cells. Both polycystins were detected on the spindle and mid-body of mitotic cells, while fibrocystin was on centrosome throughout cell cycle. Polycystins and fibrocystin may participate in regulating mucociliary sensing and transport within pulmonary airways.

Plasma generated by discharge has wide applications, which require the formation of a large-volume discharge in atmospheric pressure. The key to an optimal application performance lies in how to generate stable and diffuse plasma especially in a large-volume and in high-speed airflows1. This paper presents our study of repetitive nanosecond volume discharge behaviors in high-speed airflows.

Plasma jet is one kind of atmospheric pressure discharge which has been studied extensively. The stable /unstable states of plasma jets have been demonstrated in previous studies1. And the plasma jet which can keep in stable stage is important for application. In this study, we are focus on the influence of grounding electrode for the stage of plasma jet.

In a dielectric tube partially surrounded by distilled water, a propagating discontinuity is observed in a cold argon plasma jet at atmospheric pressure in both the laminar and turbulent flow regimes. Through electrical and optical diagnostics, the mechanism of formation for this phenomenon is investigated. Results show that the strong polarization of the distilled water, the constraint from the electric field (or a sufficiently high electric field), and the transport of the active species are in combination responsible for the propagating plasma discontinuity. Indeed, smaller surface areas of the tube surrounded by distilled water, shorter distances between the distilled water and the ground electrodes, as well as larger gas flow rates can together make the discontinuous discharge channel appear more readily. In particular, when the flow switches from laminar to turbulent, the plasma behavior of the discontinuity no longer depends on the gas flow rate. This is mostly attributed to the extreme instability of turbulence which further suppresses the transport of active species.

Persistent infection with high-risk HPV, particularly Type HPV 16 and 18, is necessary in the development of cervical cancer, but apart from HPV infection, other causative factors of most cervical cancers remain unknown. The aim of this study was to determine the prevalence of HPV 16 and HPV 18 and HSV 1 and HSV 2 in cervical samples, and to assess the role of HSVs in cervical carcinogenesis. Two hundred thirty-three healthy controls and 567 cases (333 of cervicitis, 210 of cervical intraepithelial neoplasia, and 24 of squamous cell carcinoma) in cervical exfoliative cells were tested for HPV 16, HPV 18, HSV 1, and HSV 2 DNA using the triplex real-time polymerase chain reaction method. In contrast to healthy women, positive rate of HPV is related significantly to cervical lesions (odds ratios (ORs) = 4.1, P0.01 for cervical intraepithelial neoplasia; ORs = 24.9, P0.01 for squamous cell carcinoma), but not cervicitis (ORs = 2.3, P0.05). HSV 2 prevalence in cervical intraepithelial neoplasia and squamous cell carcinoma was higher than in healthy women (ORs = 4.9, P0.05 for cervical intraepithelial neoplasia; ORs = 4.7, P0.05 for squamous cell carcinoma). HSV 2 coinfection with HPV in cervical intraepithelial neoplasia and squamous cell carcinoma was strongly higher than in healthy women (ORs = 34.2, P0.01 for cervical intraepithelial neoplasia; ORs = 61.1, P0.01 for squamous cell carcinoma). The obtained results indicated that the presence of HPV is associated closely with cervical cancer, and that HSV 2 infection or co-infection with HPV might be involved in cervical cancer development, while HSV 1 might not be involved.

Schistosomiasis is a major infectious disease and a public health concern in many areas in China and other countries. Sensitive method for detection of the parasite is critical for early diagnosis and for monitoring of effective treatment of the disease. In this study, we developed a highly sensitive TaqMan real-time PCR assay for the detection of Schistosoma japonicum DNA in mouse feces and serum samples. This assay was based on the DNA sequence of the S. japonicum 18S rRNA gene and was able to detect 10 fg of S. japonicum genomic DNA, which is 100 times more sensitive than conventional PCR. We were able to detect the S. japonicum DNA one week post-infection in mouse sera and 4 weeks post-infection in feces, which was one week earlier than egg detection by microscopy in feces. This assay was also highly specific for Asian Schistosomes which are causative species of human Schistosomiasis. In single sex male cercariae infected mice, parasite DNA was only detected in the first 4 weeks post-infection, suggesting that the DNA was derived from decaying worms’ corpse in the first 4 weeks whereas the DNA was mainly from decaying parasite eggs afterwards. Therefore we conclude that the established TaqMan real-time PCR assay is a sensitive, specific and convenient method that could be used for the early diagnostic evaluation of S. japonicum infection in humans and for monitoring outbreaks in endemic areas with low prevalence.

Human immunodeficiency virus type-1 (HIV-1) and hepatitis C virus (HCV) are transfusion-transmitted human pathogens that have a major impact on blood safety and public health. Based on multiplex asymmetrical PCR and coupled with gold labelled silver stain (GLSS), we developed the visual DNA microarray for sensitive and specific detection of these two viruses. Capturing probes of 5'-end-amino-modified oligonucleotides were immobilized on glass surface to bind the complement biotinylated target DNA. The Au-streptavidin probe was introduced to the microarray for specific binding to biotin. Black images of microarray spots which result from the precipitation of silver onto Au-streptavidin probes, were visualized by naked eyes. In order to improve the efficiency of microarray hybridization, triplex asymmetrical PCR of HIV-1, HCV and Human enterovirus 71 (EV-71, used as positive control) were performed to prepare abundant biotinylated single-stranded target DNA. The sensitivity of visual DNA microarray (10(3) copies/ml) was higher than conventional PCR (10(4) copies/ml) and was identical to FQ-PCR (10(3) copies/ml). Total 152 blood samples containing the two viruses were tested using the DNA microarray and fluorescence quantitative real-time PCR (FQ-PCR). The results were identical (P0.05). So this system has high sensitivity and may have potential in clinical applications.