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OBJECTIVE: To investigate feasibility of the quantitative parameters of dual-energy computed tomography (DECT) to assess therapy response in advanced non-small cell lung cancer (NSCLC) compared with the traditional enhanced CT parameters based on the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines. METHODS: Forty-five patients with unresectable locally advanced NSCLC who underwent DECT before and after chemotherapy or concurrent chemoradiotherapy (cCRT) were prospectively enrolled. By comparing baseline studies with follow-up, patients were divided into two groups according to RECIST guidelines as follows: disease control (DC, including partial response and stable disease) and progressive disease (PD). The diameter (D), attenuation, iodine concentration and normalized iodine concentration of arterial and venous phases (ICA, ICv, NICA, NICv) and the percentage of these changes pre- and post-therapy were measured and calculated. The Pearson correlation was used to analyze correlation between various quantitative parameters. The receiver operating characteristic (ROC) curves were used to evaluate accuracy of therapy response prediction. RESULTS: The change percentages of Attenuation (Δ-Attenuation-A and Δ-Attenuation-V), IC (ΔICA and ΔICV) and NIC (ΔNICA and ΔNICV) pre- and post-therapy correlate with the change percentage of D (ΔD). Among these, ΔICA strongly correlates with ΔD (r = 0.793, P

Transient receptor potential canonical 3 (TRPC3) is a nonselective cation channel, and its dysfunction is the basis of many clinical diseases. However, little is known about its possible role in the bladder. The purpose of this study was to explore the function and mechanism of TRPC3 in partial bladder outlet obstruction (PBOO)-induced detrusor overactivity (DO). We studied 31 adult female rats with DO induced by PBOO (the DO group) and 40 sham-operated rats (the control group). Here we report that the expression of TRPC3 in the bladder of DO rats increased significantly. Furthermore, PYR10, which can selectively inhibit the TRPC3 channel, significantly reduced bladder excitability in DO and control rats, but the decrease of the bladder excitability of DO rats was more obvious. PYR10 significantly reduced the intracellular calcium concentration in smooth muscle cells (SMCs) in DO and control rats. Finally, Na+/Ca2+ exchanger 1 (NCX1) colocalizes with TRPC3 and affects its expression and function. Collectively, these results indicate that TRPC3 plays an important role in the pathogenesis of DO through a synergistic effect with NCX1. TRPC3 and NCX1 may be new therapeutic targets for DO. The expression and functional levels of TRPC3, a nonselective cation channel, and NCX1, a Na+/Ca2+ exchanger, are increased in the bladders of rats with partial bladder outlet obstruction-induced detrusor overactivity. The synergistic effects of TRPC3 and NCX1 significantly increase the concentration of Ca2+i in smooth muscle cells, which induces bladder hyperactivity in this animal model of overactive bladder. TRPC3 and NCX1 may be new therapeutic targets for detrusor overactivity.

Qudong Lu,1 Yang Yang,1 Hengshuai Zhang,1 Cheng Chen,2 Jiang Zhao,1 Zhenxing Yang,1 Yi Fan,1 Longkun Li,1 Huan Feng,1 Jingzhen Zhu,1 Shanhong Yi1 1Department of Urology, Second Affiliated Hospital, Army Medical University, Chongqing, 400037, People’s Republic of China; 2Department of Burns, First Affiliated Hospital, Army Medical University, Chongqing, 400037, People’s Republic of ChinaCorrespondence: Jingzhen Zhu; Shanhong YiDepartment of Urology, Second Affiliated Hospital, Army Medical University, Chongqing, People’s Republic of ChinaTel +86 23-68774453Email zhujz1114@163.com; 2644057844@qq.comPurpose: Hyperalgesia and bladder overactivity are two main symptoms of interstitial cystitis/bladder pain syndrome (IC/BPS). Cannabinoid receptors participate in the modulation of pain and bladder function. GPR18, a member of the cannabinoid receptor family, also participates in the regulation of pain and bladder function, but its underlying mechanisms are unknown. In this work, we sought to study the role of GPR18 in IC/BPS.Methods: A rat model of IC/BPS was established with cyclophosphamide (CYP). Paw withdrawal threshold (PWT) measurement and cystometry were used to evaluate pain and bladder function, respectively. RT-PCR, Western blotting and immunofluorescence were used to assess the expression and distribution of GPR18. The role of GPR18 in pain and bladder function was studied by intrathecal injection of resolvin D2 (RvD2, a GPR18 agonist) and O-1918 (a GPR18 antagonist). Calcium imaging was used to study the relationship between GPR18 and TRPV1.Results: A rat model of IC/BPS, which exhibited a decreased PWT and micturition interval, was successfully established with CYP. The mRNA and protein expression of GPR18 was reduced in the bladder and dorsal root ganglia (DRG) in rats with CYP-induced cystitis. Intrathecal injection of RvD2 increased the PWT and micturition interval. However, O-1918 blocked the therapeutic effect of RvD2. GPR18 was present in bladder afferent nerves and colocalized with TRPV1 in DRG, and RvD2 decreased capsaicin-induced calcium influx in DRG.Conclusion: Activation of GPR18 by RvD2 alleviated hyperalgesia and improved bladder function, possibly by inhibiting TRPV1 in rats with CYP-induced cystitis.Keywords: interstitial cystitis/bladder pain syndrome, GPR18, TRPV1, hyperalgesia, bladder overactivity, resolvin D2, O-1918

The previous study using Sertoli cells cultured in vitro has shown that the protective effects of astragaloside IV (AsIV) on cadmium (Cd)-induced damage to Sertoli cells and its membrane proteins. Yet, it is not known if AsIV has an equivalent effect on Cd-induced damage to the spermatogenesis microenvironment in rats. Using an in vivo model, Cd-induced damage to the spermatogenesis microenvironment and the protective effects of AsIV were studied. Eighteen male Sprague Dawley (SD) rats were randomly divided into three groups (n = 6/group): Cd group, CdA Cd: cadmium; SD: Sprague Dawley; ZO-1: zonal occludin-1; Cx43: connexin 43; BTB: blood-testis barrier; MAPKs: mitogen-activated protein kinases; OSP: oligodendrocyte-specific protein; Cxs: connexins; GJIC: gap junctional intercellular communication; ROS: reactive oxygen species; MDA: malondialdehyde; TGF: tumor growth factor; PBS: phosphate buffer saline; BSA: bovine serum albumin.

The protein corona on nano drug carriers is an important well-known biological issue that often induce biological incompatibility and screens the targeting molecules on the surfaces of carriers, therefore, the design of NPs with good protein corona-free property is highly desired and challenged. The natural polysaccharide has been demonstrated as one types of stealth materials after the functional group modification process, but the types and structures of their chains has never been considered. Here, we have designed five types of core-shell starch-coated poly (methyl methacrylate) nanoparticles and we found the starch coated NPs with low amylose content (

According to the laws of thermodynamics, materials normally exhibit contraction or expansion along the directions of the applied pressure or tension. Here, we show that a man-made cocrystal of a me ...

Human urine-derived stem cells (hUSCs) show multipotential differentiation ability and can differentiate into mesodermal cell lineages. Interstitial cells of Cajal-like cells (ICC-LCs) are crucial for the pace-making function of spontaneous contraction in the bladder. However, the mechanisms by which hUSCs generate ICC-LCs have not been elucidated. In this study, we developed a strategy for directional differentiation of hUSCs into ICC-LCs. hUSCs were transfected with lentiviral vectors encoding c-Kit, stem cell factor (SCF), hyperpolarization activated cyclic nucleotide gated potassium channel 4 (HCN4), and 5-azacytidine induced 2 (AZI2) genes, and the cells were cultured for an additional 7 days in specific medium. The expression of the surface marker c-Kit on ICC-LCs was determined at 7 days after transfection. hUSCs were successfully expanded and transfected with the four lentiviral vectors. hUSCs transfected with lentiviral-c-Kit, lentiviral-HCN4, and lentiviral-AZI2 showed higher expression of c-Kit 7 days after transfection, but only the lentiviral-HCN4-transfected cells showed morphological alterations in ICC-LCs. These cells also displayed visible HCN current amplitude and density. This approach may provide a new strategy for the treatment of underactive bladder.

The protein corona on nano drug carriers is an important well-known biological issue that often induces biological incompatibility and screens the targeting molecules on the surfaces of carriers, thus causing a loss of targeting specificity. Although polyethylene glycol (PEG) and zwitterionic polymers have been widely used as anti-fouling materials, there still remain critical challenges for their use as protein-corona agents for drug delivery and targeting. Here, we have designed novel amphoteric natural starch-stabilized core-shell colloidal nanoparticles with more efficient protein corona-free properties, under long term circulation, at different protein concentrations and in different protein charge environments, compared to typical anti-fouling materials such as PEG and zwitterionic polymers. More importantly, the starch-coated polymer nanoparticles can be further functionalized by antibodies to achieve additional excellent targeting and cell internalization capabilities for their use in photodynamic therapy. Our findings demonstrate a novel protein-free or anti-fouling natural material that is very promising for use as highly efficient nano drug carriers and marine coatings.

Crystallized black phosphorus-graphite (BP-G) composites with tunable phosphorus carbon (P C) bonds have been prepared by a facile high pressure and high temperature (HPHT) method. It is found that P C bonds were formed between phosphorus and graphene layers, and their concentration can be tuned by controlling the ratio of the two precursors (i.e., red phosphorus and graphitic carbon nitride). The maximum content of P C bond is ∼13.3% in the obtained composites by HPHT treatment, which reaches the highest record reported previously by other methods. Meanwhile, by combining theoretical calculation with experiment, we studied the bonding sites of P C bonds, and also the effect of P C bonds on the cycling performance of crystallized BP-G composites, which sheds new insights into our understanding on the electrochemical performance of phosphorus/carbon materials.

Enhancement of the luminescence efficiency of up-conversion nanocrystals is essential for the development of near-infrared (NIR)-responsive photocatalysts. In this study, δ-doped hexagonal NaYF4:Yb,Tm@TiO2 (β-NaYF4:Yb,Tm@TiO2) nanocrystals, with the dopant ions Tm3+ and Yb3+ in two-dimensional planes, integrated with reduced graphene oxide (RGO) nanosheets were synthesized by a simple solvothermal process and characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), luminescence and ultraviolet absorption spectroscopy. The core, inner shell and outter shell of the δ-doped β-NaYF4:Yb,Tm are in the most efficient host structure phase, with a diameter of 29 nm and thicknesses of 6.4 and 1.2 nm, respectively. The δ-doped β-NaYF4:Yb,Tm exhibits about a 13 times increase in the luminescence intensity than the bare β-NaYF4:Yb,Tm due to the effective reduction of both the surface and inner defects around Yb3+. Moreover, the δ-doped β-NaYF4:Yb,Tm@TiO2/RGO exhibits significantly enhanced photocatalytic activity in the degradation of rhodamine B (RhB) under both NIR and UV irradiation. Our results highlight that the δ-doping construction of the most efficient host structure is promising for developing NIR-responsive photocatalysts.

The Piezo1 channel is a mechanotransduction mediator, and Piezo1 abnormalities have been linked to several clinical disorders. However, the role of the Piezo1 channel in cystitis-associated bladder dysfunction has not been documented. The current study aimed to discover the functional role of this channel in regulating bladder activity during cyclophosphamide (CYP)-induced cystitis. One hundred four female rats were randomly assigned to the control, CYP-4h, CYP-48h and CYP-8d groups. CYP successfully induced acute or chronic cystitis in these rats. CYP treatment for 48h or 8d significantly increased Piezo1 channel expression in bladder interstitial Cajal-like cells (ICC-LCs), and the increase in CYP-8d rats was more prominent. In addition, 2.5 μM Grammostola spatulata mechanotoxin 4 (GsMTx4) significantly attenuated bladder hyperactivity in CYP-8d rats by inhibiting the Piezo1 channel in bladder ICC-LCs. Furthermore, by using GsMTx4 and siRNA targeting the Piezo1 channel, we demonstrated that hypotonic stress-induced Piezo1 channel activation significantly triggered Ca2+ and Na+ influx into bladder ICC-LCs during CYP-induced chronic cystitis. In addition, the Piezo1 channel functionally interacted with the relatively activated reverse mode of Na+/Ca2+ exchanger 1 (NCX1) in bladder ICC-LCs from CYP-8d rats. In conclusion, we suggest that the functional role of the Piezo1 channel in CYP-induced chronic cystitis is based on its synergistic effects with NCX1, which can significantly enhance [Ca2+]i and result in Ca2+ overload in bladder ICC-LCs, indicating that the Piezo1 channel and NCX1 are potential novel therapeutic targets for chronic cystitis-associated bladder hyperactivity., Chronic cystitis: proteins may open a channel to treatment A protein that controls the passage of ions through cell membranes is implicated in interstitial cystitis/painful bladder syndrome (IC/PBS). This condition causes chronic pelvic pain and increased urinary frequency and urgency. Current treatment options are unsatisfactory. Researchers led by Longkun Li at the Third Military Medical University in Chongqing, China, and Mingjia Tan at the University of Michigan, Ann Arbor, USA, studied the role of this membrane channel protein, called Piezo1. Increased activity of Piezo1 was linked to bladder hyperactivity in rats with drug-induced cystitis. The research also identified a synergistic interaction between Piezo1 and a second membrane channel protein. A drug that inhibits Piezo1 activity reduced bladder hyperactivity in the rats. Drugs targeting these two proteins might help to treat the chronic cystitis of patients with IC/PBS.

Carbon nitride (C-N) materials have been attracting great interest because of their extraordinary performance in photocatalysis and energy conversion. However, developing an effective strategy for achieving band-gap engineering of C-N materials to satisfy practical applications remains highly desired. Here we report an efficient way to tune the band gap and control the nitrogen stoichiometry in carbon nitride compounds by using high pressure and high temperature (HPHT) treatment. It is found that treating a g-C3N4 precursor at relatively low temperature (630oC and below) under pressure can efficiently narrow the band gap even down to the red light region (~600 nm), increase the crystallinity, and significantly improve the charge carrier separation efficiency (by two orders of magnitude), almost without changing their stoichiometry. When increasing the treatment temperature under pressure, nitrogen-doped graphene/graphite materials with weak ferromagnetism were obtained. We thus obtained C-N materials with tunable band gaps, ranging from semiconducting to metallic states. XPS measurements show that pyridinic nitrogen is preferentially eliminated under such HPHT conditions while graphitic nitrogen is preserved in the C-N network. Our results thus provide an efficient strategy for tuning the structure and physical properties of C-N materials for applications. Supplementary information available at the journal website.

It is widely reported that fungal strains show good heavy metal ions tolerance, but the role of fungal extracellular polymeric substances (EPS) in heavy metal ion biosorption and tolerance is still not clear. In this study, a high metal ion tolerant fungal strain PTN31, identified as Aspergillus niger, was isolated, which could grow in the medium with coexistence of various metal ions. The minimum inhibitory concentration (MIC) for Pb was up to 72 mM. Moreover, biosorption behaviors and mechanisms of Pb2+ by PTN31 and extracted EPS were fully tested. Higher pH promoted biosorption of Pb2+ by the both biosorbents due to deprotonation of the functional groups in microorganisms and EPS. High and low temperature affected the physiological and metabolic activity of the biosorbents, leading to low Pb2+ adsorption. The adsorption kinetic process was well described by pseudo-second order model (R2 > 0.98) and the adsorption isotherm fit well with Freundlich model (R2 > 0.95). The maximum monolayer Pb2+ biosorption capacity of extracted EPS form PTN31 was as high as 713.6 mg/g, whereas Pb2+ biosorption capacity of PTN31 was decreased by 40.9%–66.8% after removal of EPS. Therefore, large Pb2+ biosorption capability of EPS is primarily responsible for the high lead tolerance of strain PTN31. EDS and FTIR indicated that EPS with abundant functional groups was highly related to the formation of cell aggregates, which could build a protective barrier for protecting cells from hostile environments.

Prostate cancer can progress from androgen dependence to androgen deprivation resistance with some unknown mechanisms. The current study aims to explore the possible role of pituitary tumor transforming gene1 (PTTG1) in castration-resistant prostate cancer (CRPC). Initially, we found that PTTG1 expression was significantly increased in androgen-independent prostate cancer cell lines PC3, DU145 and CRPC specimens compared with that in androgen-dependent prostate cancer cell line LNCaP and initial prostate cancer specimens. PTTG1 overexpression significantly enhanced the cell survival rate, clonality and tumorigenicity in LNCaP cells upon androgen-deprivation therapy (ADT). While knockdown of PTTG1 expression significantly elevated the sensitivity of DU145 cells to ADT. The effects of PTTG1 overexpression on LNCaP cells may be ascribed to the induced EMT and increased CD44+ CD24- cancer stem cell population. Furthermore, we detected that PTTG1 expression was regulated by interleukin-6 via activated signal transducer and activator of transcription 3 (STAT3) directly binding to the region -500 to +1 of PTTG1 promoter in LNCaP cells. In conclusion, our results elucidate that interleukin-6/STAT3 activation can increase PTTG1 expression and, consequently, promote the resistance to ADT in CRPC by inducing EMT and increasing the cancer stem cell population, suggesting that PTTG1 may be a novel therapeutic target for CRPC.

Background/Aims: Cyanidin is an anthocyanin found in many foods. Although its variable antioxidant levels are well-documented, little is known about its effects on renal cell carcinoma (RCC) tumorigenesis. This study, therefore, investigated the effects of cyanidin on the proliferation, migration, and invasion of renal cell carcinoma lines and demonstrated, for the first time, significant inhibitory effects of cyanidin on RCC tumorigenesis. Methods: RCC cells were treated with different doses of cyanidin and the effects were tested by Cell Counting Kit-8 reagent, clone formation assay, transwell assay, and flow cytometry. Moreover, the cyanidin-mediated mechanism that curtailed tumorigenesis was analyzed by RNA sequencing (RNA-seq). Sequencing data from The Cancer Genome Atlas (TCGA) were used to compare the expression of both early growth response protein 1 (EGR1) and selenoprotein W (SEPW1) in RCC and tumor-free adjacent normal tissue samples. Real-time PCR (RT-PCR) and/or western blot were used to assess the expression of E-cadherin, cleaved-caspase3, Bcl2, p62, and ATG4. Results: We found significantly greater induction of cell-cycle arrest, apoptosis, and suppression of RCC cell invasion and migration at concentrations of 25 µM and 100 µM than at a concentration of 50 µM. It was also discovered, first through RNA-seq then confirmed by RT-PCR, that cyanidin (100 µM) inhibited RCC carcinogenesis through EGR1 and SEPW1. TCGA data indicated that the expression level of EGR1 was lower and that of SEPW1 was higher in RCC tumor tissue than in normal tissues. Moreover, western blot and/or RT-PCR indicated that cleaved-caspase3 was enhanced and E-cadherin was inhibited by cyanidin treatment. Furthermore, western blot and RT-PCR also showed regulation of p62 and ATG4, which are associated with autophagy. Cyanidin in vivo significantly inhibited the growth of xenografts in nude mice. Conclusions: The results of this study showed the therapeutic potential of cyanidin for the treatment of RCC and the prevention of recurrence and metastasis.

Background Previous studies have confirmed a family risk of nephrolithiasis (NL), but only 15% of all cases are associated with an identified monogenic factor. In clinical practice, our group encountered a patient with NL combined with cystic kidney disease that had 3 affected family members. No known mutations association with NL was detected in this family, and thus further investigation of the molecular cause of NL was deemed to be necessary. Results Quality analysis from the sequencing stage showed a more than 80-fold average depth and 95% coverage for each sample, and six mutations within six genes were chosen as candidate variants for further validation. Genotyping of rs182089527in the phosphodiesterase 1A (PDE1A) gene in the validation cohort indicated that the alternative allele was present in 15 patients with heterozygosity and in 1 patient with homozygosity, and exhibited significant enrichment in NL patients (Fisher’s exact test, adjusted p = 0.0042) and kidney cystic patients (Fisher’s exact test, adjusted p = 0.067) compared to controls. In addition, function analysis displayed a significant decrease in the protein and mRNA expression levels resulting from the rs182089527 mutant sequence compared with the wild-type sequence. Moreover, patients with this mutation displayed a high level of creatinine and urea in urinalysis. Conclusions Our study provides genetic evidence that the rs182089527 mutation in PDE1A is involved in the development of NL and kidney cysts, which should help to improve personalized medicine for diagnosis and treatment. Electronic supplementary material The online version of this article (dio: 10.1186/s12864-017-4247-8) contains supplementary material, which is available to authorized users.

EP3 is a receptor for prostaglandin E2 (PGE2), and although its effect on bladder excitability has attracted considerable attention, the underlying mechanism remains unclear. To investigate whether the hyperpolarization-activated cyclic nucleotide-gated (HCN) channels in the interstitial cells of Cajal (ICCs) of the bladder are involved in the effect of EP3 activation on bladder excitability, wild-type mice, HCN1 knockout (HCN1-/-) mice and rats were used in our study. Double immunofluorescence staining and immunoprecipitation assays demonstrated the interaction between EP3 and the HCN channels. Sulprostone is a selective agonist of EP3. The current density of HCN channels was enhanced by sulprostone or PGE2 using whole-cell patch clamping. Western blot analyses showed that the expression levels of HCN1 and HCN4 were higher in bladders that had undergone intravesical instillation with sulprostone than in bladders treated with normal saline (NS). Both PGE2 and sulprostone increased the calcium concentration of the ICCs, and their effects were inhibited by ZD7288 (antagonist of HCN channels) treatment. In bladder detrusor strip testing, both PGE2 and sulprostone enhanced the amplitude of the bladder detrusor in HCN1-/- mice; however, these effects were less than those in the wild-type mice. Furthermore, the effects of PGE2 and sulprostone were inhibited by ZD7288. Taken together, our results indicate that EP3 is expressed in bladder ICCs and facilitates bladder excitability via HCN channels. This study provides more comprehensive insights into the mechanism between inflammation and bladder excitability and highlights methods that can resolve bladder hyperactivity.

Calcium-activated nucleotidase 1 (CANT1, belongs to the apyrase family, is widely expressed in various organs. However, the biological function of CANT1 remains poorly explored. In this study, we aimed to investigate the expression profile and functions of CANT1 in clear cell renal cell carcinoma (ccRCC). Our data show that the protein level of CANT1 was significantly higher in tumor tissues than in adjacent normal tissues. CANT1 silencing suppressed cell proliferation, migration, and invasion obviously in 769-P and 786-O cells, arrested cell cycle in S phase and promoted apoptosis in 769-P cells. In conclusion, the present study shows the different expression mode of CANT1 in human ccRCC tumor tissue and adjacent normal tissue, denotes the function of CANT1 in ccRCC cells and provides potential molecular mechanisms and pathways of CANT1 antitumor function in ccRCC.

Graphitic carbon nitride (C2N and C3N) with various π electron distributions on layers have been studied under pressure through a combined theoretical and experimental approach and a comparison with graphite. It is found that as these materials transform into low compressibility phases in the pressure range from 15 to 45 GPa, strong electrostatic repulsion between π electrons and in-plane sp2 electrons may distort and soften the sp2 bonds, leading to anomalous pressure evolutions of the intralayer phonon vibrations, such as a plateau-like behavior of E2g mode (G-band) in C2N and C3N. This also causes a slow increase in the resistivity/resistance of C2N and C3N as pressure increases, and the gradual interlayer bonding leads to an abrupt increase in resistance of the materials but with different pressure responses due to their different π electron distributions. Moreover, the intensity enhancement of the G band in both CN materials may be related to their electronic structure changes. The results deepen our understanding of the effects of π electron distribution on the structural transition of graphitic materials and may explain some unexplained in previous studies.

Porous graphitic carbon nitride (g-C3N4) with tunable nitrogen vacancies was prepared by a high pressure and high temperature (HPHT) process combined with hard template method. The concentrations of two-coordinated nitrogen vacancy can be tuned by temperatures that are much gentler than those required at ambient pressure. Such produced samples after HPHT treatment show decreased interlayer distance, up-shift in valance band (VB) and down-shift in conduction band (CB), and thus exhibit narrowed bandgap down to 1.88 eV, as well as increased efficiency of charge carrier separation and transfer. Consequently, the porous vacancy-rich g-C3N4 with increased specific surface area exhibits 13.5 times enhancement in photocatalytic H2 evolution compared to the bulk g-C3N4. The underlying mechanism for the significantly enhanced photocatalytic H2 production is further proposed. This process is efficient for defects engineering of carbon nitride materials with simultaneous realization of porous structure, which might be extended to other materials for superior performance.

Preventing the high carrier recombination rate of graphitized C3N4 (GCN) is an urgent problem to be solved for its application as a photocatalyst for hydrogen production. Here, we first rationally embed the benzene ring in GCN to modify the local symmetry without changing its long-order structure. Theoretical calculation predicts that this design can change the electronic structure and promote the effective charge transfer in GCN. The benzene ring embedded GCN is successfully prepared by copolymerization using dicyandiamide and terephthalonitrile as precursors. Photoluminescence (PL) and time-resolved transient PL (TRPL) spectra confirm that the electron transfer efficiency of the benzene ring embed GCN is greatly improved. This nanomaterial displays 10.8 times higher photocatalytic hydrogen production rate than that of pristine GCN with the apparent quantum yield of 11.3% at 400 nm and 9% at 420 nm. This work provides a novel strategy for designing high-efficiency two-dimensional (2D) photocatalytic materials for water splitting.

A thermostable GDSL family esterase-encoding gene, EstL5, was directly obtained from the genomic DNA of Geobacillus thermodenitrificans T2. Recombinant hexahistidine-tagged EstL5 was overexpressed, purified, and its biochemical properties were partially characterized. EstL5 was observed to be active within the temperature range of 0–80°C, having maximal activity at 60°C. Unlike most other thermostable enzymes, EstL5 displayed 24% of its highest activity at 0°C. EstL5 exhibited a high level of activity within a pH range of 6.0–11.0, showing the highest activity at pH 8.0. EstL5 also retained 100% of its activity after a 12-h incubation at 55°C. Furthermore, this enzyme was observed to be strongly inhibited by 10% (w/v) SDS and 0.1 mM PMSF.

Objective To uncover the role of c-Jun, a proto-oncogene, in inhibitory effects of antiproliferative factor (APF) on tumor cell growth. Materials and Methods Expression of c-Jun was analyzed by Western blotting in 45 clinical specimens (30 tumorous tissues and 15 paired non-tumorous tissues) and 3 bladder cancer cell lines. APF-responsive T24 transitional carcinoma bladder cells were treated with APF or mock control. Cell proliferation was measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. Change of c-Jun expression was detected by RT-PCR and Western blotting. The influence of c-Jun on APF treatment was evaluated by transient transfection of c-Jun and MTT assay in T24 cells. Results c-Jun was significantly higher in invasive bladder cancer tissues and cell lines. T24 cells treated with APF had decreased c-Jun expression and suppressed cell growth. More importantly, ectopic c-Jun attenuated APF inhibitory effects on cell growth. Conclusions These observations suggest that c-Jun is involved in APF-mediated inhibition for bladder tumor cell growth, as potential target of APF in patients with aggressive bladder carcinoma.

Long non-coding RNA plasmacytoma variant translocation 1 (PVT1) is up-regulated in various human cancers, and our results indicated that PVT1 was up-regulated in clear cell renal cell carcinoma tissues. The Cancer Genome Atlas cohort analysis revealed that in clear cell renal cell carcinoma, higher PVT1 expression correlated with advanced TNM stage, histological grade, and poor survival. PVT1 knockdown promoted apoptosis, inhibited renal cancer cell proliferation, decreased Mcl-1, and increased cleaved caspase-3 and cleaved PARP. PVT1 increased Mcl-1 mRNA levels in renal cancer cells by promoting mRNA stability without influencing its transcription. in vitro, the enhanced apoptosis arising from PVT1 suppression was attenuated by overexpressing Mcl-1. In addition, in vivo experiments showed that PVT1 knockdown repressed xenograft tumor growth, while Mcl-1 overexpression partially rescued xenograft tumor growth. These results indicate the PVT1/Mcl-1 pathway inhibits renal cancer cell apoptosis in vitro and in vivo. PVT1 may thus serve as a novel biomarker, and the PVT1/Mcl-1 pathway may be a useful therapeutic target for clear cell renal cell carcinoma.

Although DBP (di-n-butyl phthalate) is commonly encountered as an artificially-synthesized plasticizer with potential to impair fertility, we confirm that it can also be biosynthesized as microbial secondary metabolites from naturally occurring filamentous fungi strains cultured either in an artificial medium or natural water. Using the excreted crude enzyme from the fungi for catalyzing a variety of substrates, we found that the fungal generation of DBP was largely through shikimic acid pathway, which was assembled by phthalic acid with butyl alcohol through esterification. The DBP production ability of the fungi was primarily influenced by fungal spore density and incubation temperature. This study indicates an important alternative natural waterborne source of DBP in addition to artificial synthesis, which implied fungal contribution must be highlighted for future source control and risk management of DBP.

CaF2 with different morphologies and hierarchical structures was prepared under microwave-assisted condition by a simple hydrothermal route using CaCl2 and ionic liquid [BMIM]BF4 as initial reagents, respectively. It was found that both the precursor and pH values played an important role in the formation of CaF2 crystals with different morphologies. The results suggested that the CaF2 with cube and spherical structures was obtained at different pH values (4, 8, 11), while the CaF2 with dendrite shape was formed through an oriented self-assembly growth using (NH4)2HPO4 as a structure directing agent. Scanning electron microscope observation showed that the as-prepared CaF2 was of three-dimensional eight-horn-shaped dendritic structure. The influence of the pH values for the reaction solution and precursors has been discussed in detail. On the basis of the experimental results, possible growth mechanisms of the CaF2 microcrystals were proposed.

The photo-assisted epoxidation of styrene over V/SBA-15 modified by alkaline-earth metal ions was investigated with hydrogen peroxide at room temperature. The results indicated that the isolated VO4 tetrahedron dispersed on the SiO2 was the photo-assisted catalytic active site for the formation of styrene oxide, while V5+ polymer species in V/SBA-15 with highly loaded V species caused deep oxidation of styrene oxide. Among the catalysts modified by alkaline-earth metal ions, Mg–V/SBA-15 displayed the best catalytic performance. It was probably due to the fact that the metallic character of Mg was weaker in alkaline-earth metals, so it was easier to disperse V5+ polymer species and the dispersion effect was especially obvious in the comparatively high loading weight of vanadium oxide. Moreover, addition of alkaline-earth metals decreased the acidic sites of V/SBA-15, inhibiting the ring-opening reaction on which the deep oxidation occurred. The structures of all catalysts were thoroughly investigated using different characterization techniques, including X-ray diffraction (XRD), transmission electron microscopy (TEM), NH3-temperature programmed desorption (TPD), H2 temperature-programmed reduction (TPR) and X-ray photoelectron spectroscopy (XPS), etc. A possible catalytic mechanism was proposed.

UBE1 plays an important role in the first step of ubiquitin-proteasome pathway to activate ubiquitin. Both the structure and biochemical property research of human UBE1 protein, and the activity analysis of those enzymes which are related with ubiquitination pathway, are based on high purity of UBE1 protein. To obtain human UBE1 protein, the full length of human UBE1 was expressed in E. coli and purified by Ni-NTA superflow sepharose and strep-tactin sepharose which based on UB-UBE1 high-energy thioester bonded intermediate complex. It was demonstrated that purified UBE1 could activate and conjugate UB to ubiquitin-conjugating enzyme E2s. The purified large amount of UBE1 could be used for in vitro studies of ubiquitin pathway and structural studies.

Tungstophosphoric acid (H3PW12O40) (PW) catalysts supported on mesoporous Mn-MCM-41(n Si/n Mn = 25) with various (10, 20 and 30 wt%) acid loadings were prepared by impregnation, and their physical chemical properties were characterized by powder X-ray diffraction (XRD), N2 adsorption–desorption, Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), high resolution transmission electron microscopy (HRTEM), and X-Ray Photoelectron Spectroscopy (XPS). The host material suffers severe structural distortions at higher loading, which lead to a considerable loss of long-range orders. Their catalytic performances were evaluated in the epoxidation of styrene with dilute H2O2 (30%) as an oxidizing agent under liquid phase reaction conditions over PW/Mn-MCM-41 catalysts for selective synthesis of styrene oxide. The influences of various reaction parameters such as temperature, time, solvents, PW loading and catalyst amount, styrene to H2O2 mmol ratios and acetonitrile (MeCN) to N,N-dimethylformamide (DMF) volume ratios on the conversion of styrene and yield as well as selectivity of styrene oxide have also been studied in details. The results revealed that 20% PW/Mn-MCM-41 (n Si/n Mn = 25) was more active than other catalysts. The PW/Mn-MCM-41, moreover, was found to be reusable and environmentally benign for the epoxidation of styrene.

Most members of the type-2 phosphatidic acid phosphatase (PAP2) superfamily are integral membrane phophatases involved in lipid-related signal transduction and metabolism. Here we describe the cloning of a novel gene from Geobacillus toebii T-85, encoding a PAP2-like protein, Gtb PAP2L2, which contains 212 amino acids and shows a limited homology to other known PAP2s, especially at conserved phosphatase motifs, and a similar six-transmembrane topology structure. This enzyme was expressed, and purified in Escherichia coli. Recombinant Gtb PAP2L2s from the membrane fractions were solublized with 0.3% (v/v) Triton X-100 and purified by Ni(2+) affinity chromatography. The purified enzyme showed broad substrate specificity to phosphatidic acid, diacylglycerol pyrophosphate, and lysophosphatidic, but preferred phosphatidic acid and diacylglycerol pyrophosphate in vitro. Gtb PAP2L2 is a thermal stable enzyme with a half-life of 30 min at 60 degrees C. The enzyme was strongly inhibited by 1% SDS, 10 mM veranda, and Zn(2+), whereas it was independent of the Mg(2+) ion, and insensitive to N-ethylmaleimide. The purified recombinant Gtb PAP2L2 was catalytically active and highly stable, making it ideal as a candidate on which to base further PAP2 structure/function studies.

Mesoporous Ti-MCM-41 molecular sieve was synthesized under hydrothermal conditions. 5, 10, 20, 30 and 50 wt.% phosphotungstic acid (H 3 PW 12 O 40 ) (PW) were supported on Ti-MCM-41 and the resulting catalysts were characterized by powder X-ray diffraction (XRD), N 2 adsorption–desorption, Fourier transform infrared spectroscopy (FT-IR), UV–vis diffuse reflectance spectroscopy (DRS), thermogravimetric analysis (TGA) and differential thermal analysis (DTA), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and high-resolution transmission electron microscopy (HRTEM). The host material suffers severe structural distortions at higher loading, which lead to a considerable loss of long-range orders. Their catalytic performances were evaluated in the epoxidation of styrene with dilute H 2 O 2 (30%) as an oxidizing agent under liquid-phase reaction conditions over PW/Ti-MCM-41 catalysts for selective synthesis of styrene oxide. The influences of various reaction parameters such as temperature, time, solvents, PW loading and catalyst amount, styrene to H 2 O 2 mmol ratios and acetonitrile (MeCN) to N , N -dimethylformamide (DMF) volume ratios on the conversion of styrene and yield as well as selectivity of styrene oxide have also been studied in details. The results revealed that 20% PW/Ti-MCM-41 was more active than other catalysts. The PW/Ti-MCM-41, moreover, was found to be reusable and effective for the epoxidation of styrene.

Ubiquitin and ubiquitin-like proteins are known to be covalently conjugated to a variety of cellular substrates via a three-step enzymatic pathway. These modifications lead to the degradation of substrates or change its functional status. The ubiquitin-activating enzyme (E1) plays a key role in the first step of ubiquitination pathway to activate ubiquitin or ubiquitin-like proteins. Ubiquitin-activating enzyme E1-domain containing 1 (UBE1DC1) had been proved to activate an ubiquitin-like protein, ubiquitin-fold modifier 1 (Ufm1), by forming a high-energy thioester bond. In this report, UBE1DC1 is proved to activate another ubiquitin-like protein, SUMO2, besides Ufm1, both in vitro and in vivo by immunological analysis. It indicated that UBE1DC1 could activate two different ubiquitin-like proteins, SUMO2 and Ufm1, which have no significant similarity with each other. Subcellular localization in AD293 cells revealed that UBE1DC1 was especially distributed in the cytoplasm; whereas UBE1DC1 was mainly distributed in the nucleus when was cotransfected with SUMO2. It presumed that UBE1DC1 greatly activated SUMO2 in the nucleus or transferred activated-SUMO2 to nucleus after it conjugated SUMO2 in the cytoplasm.

A gene-encoding alkaline phosphatase (AP) from thermophilic Geobacillus thermodenitrificans T2, termed Gtd AP, was cloned and sequenced. The deduced Gtd AP protein comprises 424 amino acids and shares a low homology with other known AP (35% identity), while it exhibits the conservation of the active site and structure element of Escherichia coli AP. The Gtd AP protein, without a predicted signal peptide of 30 amino acids, was successfully overexpressed in E. coli and purified as a hexa-His-tagged fusion protein. The pH and temperature optima for purified enzyme are 9.0 and 65 degrees C, respectively. The enzyme retained a high activity at 45-60 degrees C, while it could be quickly inactivated by a heat treatment at 80 degrees C for 15 min, exhibiting a half-life of 8 min at 70 degrees C. The K(m) and V (max) for pNPP were determined to be 31.5 muM and 430 muM/min at optimal conditions. A divalent cation is essential, with a combination of Mg2+ and Co2+ or Zn2+ preferred. The enzyme was strongly inhibited by 10 mM ethylenediaminetetraacetic acid (EDTA) and vanadate but highly resistant to urea and dithiothreitol. The properties of Gtd AP make it suitable for application in molecular cloning or amplification.

Uracil phosphoribosyltransferase, which catalyzes the conversion of uracil and 5-phosphoribosyl-1-R-diphosphate to uridine monophosphate, is important in the pyrimidine salvage pathway and is an attractive target for rational drug design by incorporation of prodrugs that are lethal to many parasitic organisms specifically. So far, uracil phosphoribosyltransferase has been reported in Arabidopsis thaliana only, not in mammals. In this study, a novel uracil phosphoribosyltransferase family cDNA encoding a 309 amino acid protein with a putative uracil phosphoribosyltransferase domain was isolated from the human fetal brain library. It was named human UPRTase (uracil phosphoribosyltransferase). The ORF of human UPRTase gene was cloned into pQE30 and expressed in Escherichia coli M15. The protein was purified by Ni-NTA affinity chromatography, but UPRTase activity could not be detected by spectrophotometry. RT-PCR analysis showed that human UPRTase was strongly expressed in blood leukocytes, liver, spleen, and thymus, with lower levels of expression in the prostate, heart, brain, lung, and skeletal muscle. Subcellular location of UPRTase-EGFP fusion protein revealed that human UPRTase was distributed in the nucleus and cytoplasm of AD293 cells. Evolutional tree analyses of UPRTases or UPRTase-domain-containing proteins showed that UPRTase was conserved in organisms. UPRTases of archaebacteria or eubacterium had UPRTase activity whereas those higher than Caenorhabditis elegans, which lacked two amino acids in the uracil-binding region, had no UPRTase activity. This means that human UPRTase may have enzymatic activity with another, unknown, factor or have other activity in pyrimidine metabolism.

Copper is an essential heavy metal trace element that plays important roles in cell physiology. The Cut family was associated with the copper homeostasis and involved in several important metabolisms, such as uptake, storage, delivery, and efflux of copper. In this study, a novel Cut family cDNA was isolated from the human fetal brain library, which encodes a 273 amino acid protein with a molecular mass of about 29.3 kDa and a calculated pI of 8.17. It was named hCutC (human copper transporter protein CutC). The ORF of hCutC gene was cloned into pQE30 vector and expressed in Escherichia coli M15. The secreted hCutC protein was purified to a homogenicity of 95% by using the Ni-NTA affinity chromatography. RT-PCR analysis showed that the hCutC gene expressed extensively in human tissues. Subcellular location analysis of hCutC-EGFP fusion protein revealed that hCutC was distributed to cytoplasm of COS-7 cells, and both cytoplasm and nucleus of AD293 cells. The results suggest that hCutC may be one shuttle protein and play important roles in intracellular copper trafficking.

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are confirmed to be expressed in bladder interstitial Cajal-like cells (ICC-LCs), but little is known about their possible role in cystitis-associated bladder dysfunction. The present study aimed to determine the functional role of HCN channels in regulating bladder function under inflammatory conditions. Sixty female wild-type C57BL/6J mice and sixty female HCN1-knockout mice were randomly assigned to experimental and control groups, respectively. Cyclophosphamide (CYP)-induced cystitis models were successfully established in these mice. CYP treatment significantly enhanced HCN channel protein expression and Ih density and significantly altered bladder HCN1 channel regulatory proteins. Carbachol (CCH) and forskolin (FSK) exerted significant effects on bladder ICC-LC [Ca2+]i in CYP-treated wild-type (WT) mice, and HCN1 channel ablation significantly decreased the effects of CCH and FSK on bladder ICC-LC [Ca2+]i in both naive and CYP-treated mice. CYP treatment significantly potentiated the spontaneous contractions and CCH (0.001–10 μM)-induced phasic contractions of detrusor strips, and HCN1 channel deletion significantly abated such effects. Finally, we demonstrated that the development of CYP-induced bladder overactivity was reversed in HCN1−/− mice. Taken together, our results suggest that CYP-induced enhancements of HCN1 channel expression and function in bladder ICC-LCs are essential for cystitis-associated bladder hyperactivity development, indicating that the HCN1 channel may be a novel therapeutic target for managing bladder hyperactivity. Research implicating a protein channel in bladder cell membranes in inflammatory bladder disorders could lead to new treatments. Longkun Li and colleagues at the Third Military Medical University in Chongqing, China, investigated a specific type of bladder cell in mice with chemically-induced cystitis, an inflammation of the bladder. Their results suggest that the cystitis was linked to increased production of the membrane protein HCN, which allows ions to pass through cell membranes. The cells concerned, known as interstitial Cajal-like cells, have already been implicated in various bladder disorders in humans, but the mechanism of their involvement was unclear. If their role in mice can also be confirmed in humans, drugs that interact with the HCN protein, or reduce its production, may be able to treat cystitis and other inflammatory bladder disorders.

The novel thermostable esterase EstL5 belonging to the GDSL family exhibits a unique cold-adaptation feature at low temperatures. To better understand its biochemical and enzymatic properties, recombinant EstL5 protein was purified and crystallized using the vapour-diffusion method. The EstL5 crystals diffracted X-rays to 2.79 A resolution using a synchrotron-radiation source, belonged to the tetragonal space group P41212 or P43212, with unit-cell parameters a = b = 101.51, c = 124.22 A, and are expected to contain two molecules in each asymmetric unit. To obtain initial phases, selenomethionyl-substituted protein was overproduced. Purified SeMet-labelled EstL5 protein was crystallized and formed crystals that diffracted to a resolution of 3.0 A.

The Escherichia coli cyclic AMP receptor protein (CRP) is a well known transcription activator protein. In this study, CRP was overexpressed, purified and cocrystallized with cAMP and a 38 bp full-length double-stranded DNA fragment. The full-length segment differed from the half-site fragments used in previous crystallization experiments and is more similar to the environment in vivo. CRP-cAMP-DNA crystals were obtained and diffracted to 2.9 Å resolution. The crystals belonged to space group P3121, with unit-cell parameters a = b = 76.03, c = 144.00 Å. The asymmetric unit was found to contain one protein molecule and half a 38 bp full-length double-stranded DNA fragment, with a Matthews coefficient of 2.62 Å(3) Da(-1) and a solvent content of 53.14%.