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The Shugan Hewei recipe (SHR) is a well-recognized traditional Chinese medicine (TCM) prescription that has been shown to significantly improve chest pain, acid regurgitation, and the mood of GERD. Nonetheless, the underlying mechanisms remain unclear. In this study, the active compounds and targets of SHR were predicted using network pharmacology. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were utilized to explore the therapeutic mechanism of SHR. Combined with the drug target obtained from network pharmacology, the therapeutic effect and mechanism of SHR were observed. SHR’s main active compounds included quercetin, kaempferol, and luteolin. The core targets of SHR and GERD were TGF-β1, IL-1β, IL-4, CXCL10, MAPK1, MAPK3, CXCL8, IL-10, IL-2, and FOS, involving virus infection, inflammatory response, and body immunity. The core targets of SHR during the treatment of mental disorders were GABRA1, GABRA2, GABRA3, GABRA5, and GABRA6, involving synaptic transmission and transmembrane movement. Animal experiments revealed that SHR could repair the lower esophageal mucosa, mediate inflammatory factors, and GABA receptors and improve the behavior of rats. Overall, our results substantiate that SHR has huge prospects for widespread application in treating GERD subjects with anxiety and depression.

In the field of cultural relic protection, the migration of moisture will cause a series of adverse effects, such as disruption and crater eruption. Assessment of water migration status is the key to early warning and prevention of these problems. In an attempt for a solution, the whole process of moisture migration in rammed earth is studied by using the moisture migration initiation criterion based on entropy. We analyzed and compared the entropy change law of the infrared radiation temperature in the two main migration stages, and we made a judgment on whether the moisture state is in the steady equilibrium or in the process of dynamic migration. The results show that the moisture migration status can be identified quickly and in real time by using the moisture migration initiation criterion based on entropy. The results of this study provide a new method for judging the state of moisture migration in geotechnical engineering, especially in the field of cultural relic protection, which has very important practical significance.

A novel porous core-shell magnetic β-cyclodextrin/graphitic carbon nitride photocatalyst (Mβ-CD/GCN) was synthesized and employed in a solar light driven catalytic system for the degradation of polychlorinated biphenyls (PCBs). The Mβ-CD/GCN display superior photocatalytic performance on account of porous structure and ultrathin GCN nanosheets design, the former improves the utilization of visible light by multiple scattering and reflection of incident light, and the latter accelerates electron transfer. The ultrahigh specific surface area (1255 m

Herein, we found that serum chemokine ligand 14 (CXCL14) was significantly enhanced in patients with idiopathic pulmonary fibrosis (IPF). In our current study, mouse L929 fibroblasts were stimulated with lipopolysaccharide (LPS) (100 ng/mL). Cell proliferation, the levels of matrix metalloproteinase 2 (MMP2) and MMP9, as well as extracellular matrix (ECM) content were assessed to evaluate the fibrogenesis of L929 cells. Proliferating cell nuclear antigen and cell viability were assessed to evaluate cell proliferation. Hydroxyproline (Hyp), collagen I/III, connective tissue growth factor (CTGF), and phosphorylated Smad2/3 (p-Smad2/3) were assessed to evaluate ECM secretion and deposition. α-Smooth muscle actin (α-SMA) was used to measure the occurrence of differentiation from fibroblast toward myofibroblast. Our data suggested that knockdown of CXCL14 prevented LPS-induced fibrogenesis of L929 cells through inhibiting cell proliferation and decreasing the expression of MMP2/9, Hyp, collagen I/III, CTGF, p-Smad2/3, and α-SMA. Notably, upregulation of protein phosphatase magnesium-dependent 1A (PPM1A) was involved in this process. On the contrary, recombinant CXCL14 protein led to an opposite effect. We first suggested that overexpression of PPM1A ameliorated LPS-induced fibrogenesis. Furthermore, we substantiated that knockdown of CXCL14 exerted an antifibrotic effect in IPF in vitro probably via the upregulation of PPM1A. Besides, evidently enhanced CXCL14, yet reduced PPM1A, was found in bleomycin-induced rat pulmonary fibrosis, confirming the roles of CXCL14 and its potential association with PPM1A in IPF in vivo. In conclusion, CXCL14 could be considered as a therapeutic target for preventing fibrogenesis of mouse L929 fibroblasts.

In this work, we have developed a novel pneumatic microfluidic chip (pChip) for in-vivo antibacterials screening at single-organism resolution using a P. aeruginosa-C. elegans infection model. To improve the accuracy and efficiency of nematode infection-based drug screening, we designed a pneumatic-controlled micro-sampler that enables simultaneous dispensing of a large number of individual free-moving worms into each culture chamber under physiological conditions without any mechanical squeezing, and the average probability of a single worm in a chamber is 88.5%, with RSD of 2.8%. Furthermore, we proposed an effective liquid P. aeruginosa-C. elegans infection model based on microfluidic chip, and the performance of the platform was verified with multiple clinical antibiotics. By applying this developed worm pChip to evaluate the antimicrobial activity of some natural medicinal compounds (coptisine, baicalin and gypenoside), we found that gypenoside can increase the P. aeruginosa infected worm survival by 2.3-fold at 30.0 μg/mL, whereas it does not suppress pathogen growth in vitro. It indicates that the well-established pneumatic chip will be an efficient platform for evaluation of antibacterial potential of medicinal natural compounds.

Karyotype is the most important diagnostic and prognostic parameter in myelodys-plastic syndrome (MDS). Here, we describe a novel case of MDS with complex chromosomal abnormalities.A 55-year-old Chinese female was admitted to the hospital for facial edema and a loss of appetite. Bone marrow aspiration showed the blast cell count 3.6%. Erythrocyte hyperplasia was active, megaloblastoid change was observed, and a wide variability of nuclear numbers, as well as variability of size and shape was present. Bone marrow chromosomal analyses showed 45~48, X, -X, -4, t (5;8) (q13;q22), add (7) (q11), add (13) (p11), -14, del (16) (p13), add (19) (q13), -20, i(21)(q10),+4~6mar [cp15]/46,XX[5]. The patient was diagnosed with MDS with WPSS of the high risk group. IPSS was medium risk-2. IPSS-R was categorized as the extremely high risk group.The prognosis and treatment of MDS with complex chromosomal abnormalities are still uncertain, and further studies are needed.

In view of the large deformation of thin-layer soft rock in the No. 2 inclined shaft of the Muzhailing Tunnel, we performed an experimental investigation on the mineral composition, physical characteristics, and uniaxial compressive strength of the surrounding rock of the tunnel. The characteristics of the large deformation of the surrounding rock of the tunnel were analyzed, and the main factors influencing the deformation of the tunnel were revealed. The influence of various factors on the large deformation of the surrounding rock was analyzed using the 3DEC-Trigon discrete element numerical simulation method. The results show that (1) the deformation of the surrounding rock of the tunnel has remarkable asymmetry, the deformation of the initial support of the tunnel is significant, and the buried depth of the area where the maximum deformation of the tunnel exceeded 1 m is greater than 500 m; (2) the main factors influencing the deformation of a thin-layer slate tunnel include joint inclination, buried depth, water absorption, and softening of the surrounding rock; and (3) the maximum deformation of the surrounding rock is observed for a joint angle of 45°, at which the buried depth is directly proportional to the deformation and failure of the tunnel. Furthermore, after the surrounding rock was softened by water absorption, the floor of the tunnel, the left shoulder socket, and the right side of the tunnel are deformed greatly. The results of this study will provide a theoretical basis for the study of similar deformation control methods and supporting measures for tunnels excavated in thin-layer soft rock.

A simple and efficient access to arylboronates was achieved with methanol-initiated borylation of aryldiazonium salts. Reduction of aryldiazonium ions by single electron transfer from methanol affords aryl radical species, which undergo a subsequent C–B bond formation with bis(pinacolato)diboron. This highly practical borylation process, which can be carried out on the gram-scale, enjoys operational simplicity as well as mild and catalyst-free conditions.

In order to solve the problem of quantitative description of water migration in a stone artifacts site, we have investigated the dynamic migration process of water and established a quantitative descriptive theory system based on infrared radiation temperature through the infrared thermal imaging method. We propose the criterion of water movement initiation based on entropy theory. A quantitative relationship equation between infrared radiation temperature and water content is established using the first law of thermodynamics. A method of identifying the moisture migration wetting front is proposed. The above theory is verified by laboratory experimental thermal imaging results of moisture migration of siltstone from the Mogao Grottoes cliff. Therefore, our method can quantitatively describe the start and migration of water at a rock site using our theoretical system and infrared thermography in a non-destructive and real-time manner. Here, we provide a new method for quantitative description of moisture migration in a stone artifacts site.

Shale is a fine-grained, clastic sedimentary rock composed of clay minerals and other minerals. The water-shale interaction plays an important role in the borehole drilling and hydraulic fracturing during the process of shale gas exploitation. In this study, laboratory experiments, including water absorption experiments and uniaxial compression tests, were used to investigate the mechanical behaviours of the water-absorbing shale specimens. From the laboratory experiments, the following statements can be drawn: a) the water-absorbing shale specimens experience longer compaction and crack closure stages than the natural shale specimen; b) the mechanical properties of the water-absorbing shale specimens, such as uniaxial compressive strength, Young's modulus and peak strain, are less than those of the natural shale specimens; c) the apertures of the failure cracks in the water-absorbing shale specimens are greater than those in the natural shale specimens under uniaxial compression loading. The acoustic emission technique was also used to capture the acoustic emission events of the shale specimens during the uniaxial compression tests. By the acoustic emission analysis, it is concluded that the water absorption stimulates the AE events in the shale specimens.

Pulmonary fibrosis (PF) is a heterogeneous pathological process in lung tissues with a considerable mortality rate. Currently, combination therapy represents an effective approach to treat PF. Dexamethasone (Dxs) and berberine (BBR) are widely applied to inhibit the progression of PF. Dxs plus penehyclidine hydrochloride or alfacalcidol have been reported more effective in therapy compared with any single drug treatment. However, whether Dxs plus BBR induces an increased antifibrotic effect remains unknown. The current study aimed to evaluate the therapeutic effect of BBR plus Dxs in bleomycin (BLM)-induced PF. A PF model in rats was established and rats were divided into control, BLM, BBR, Dxs and BBR plus Dxs groups (n=9/group). On days 3, 7 and 14, blood samples were collected from the eyes of the rats (n=6/group). CXC chemokine ligand 14 (CXCL14), collagen I, collagen III, matrix metalloproteinase (MMP)2 and MMP9 serum levels were measured by ELISA. On day 14, all rats were sacrificed. Hematoxylin and eosin analysis, Masson staining and hydroxyproline (Hyp) assessment were performed to observe histopathological changes and collagen deposition. mRNA and protein levels of CXCL14, CXC chemokine receptor 4 (CXCR4), collagen I/III, α-smooth muscle actin (α-SMA), MMP2/9 and phosphorylated-Smad 2/3 in lung tissue were further evaluated. Similar effects in preventing lung damage were observed histopathologically for Dxs and BBR compared with the BLM group. These treatments further reduced levels of Hyp, CXCL14, CXCR4, collagen I/III, MMP2/9, α-SMA and p-Smad 2/3. The combination of Dxs and BBR exhibited increased effectiveness compared with the single treatments. Results further suggested that antifibrotic mechanisms were involved in inhibiting CXCL14 and MMP2/MMP9 expression, and preventing the activation of Smad2/3 and hedgehog signaling pathways. The combined use of Dxs and BBR may represent a potential therapeutic approach for PF.

In geotechnical engineering, the water saturation of rock cannot be obtained in real time and is lossless. To solve this problem, a continuous classification representation method of water saturation of rock is proposed herein, the classification and recognition theory of the water saturation level of rock is established, and near-infrared spectrum acquisition experiments of rock under different water saturation levels are carried out. Based on the near-infrared spectrum, the partial least square (PLS) method is used to establish the recognition model, and the model is applied to the real-time identification of the saturation level in the gravel water absorption process. The results are as follows: (1) In this paper, the method of hierarchical representation of water saturation of rock is proposed, which solves the limited extrapolation ability and extrapolation precision problems of the preparation accuracy of water saturation of the rock sample and the classification-learning algorithm and thus provides the completeness and feasibility for the identification of water saturation of rock. (2) The classification recognition theory and method based on near-infrared spectroscopy to set up the water saturation level of rock have better recognition precision and can realize the water saturation level of the rock in real time and nondestructively, and (3) when the PLS method is used to establish the recognition model, the appropriate threshold parameters are selected to eliminate the abnormal samples, and multiple spectral segments are used in the modeling, which can greatly improve the recognition accuracy of the model.

Aims To investigate the effect and underlying mechanism of melittin and tripartite motif (TRIM) family in human embryonic lung fibroblast (HELF). Materials and methods Lentiviral RNA interference vector and lentiviral overexpression vector were constructed and packaged by transfecting 293T cells; the proliferation of HELF was examined using Cell Counting Kit 8; Western blot and qRT-PCR were performed to examine protein and mRNA expression; the interaction with protein phosphatase magnesium-dependent 1A (PPM1A) was examined by Co-immunoprecipitation. Key findings Compared with the control group, the mRNA expression of the TRIM6, TRIM8 and TRIM47 in the IPF group significantly increased. Melittin inhibited the mRNA expression and protein expression levels of TRIM47, the HELF proliferation, the hydroxyproline levels, and the phosphorylation of Smad2/3; the interference of TRIM47 inhibited the protein expression of Vimentin, α-SMA, CTGF, the phosphorylation of Smad2/3 and the synthesis of hydroxyproline; TRIM47 overexpression elevated the phosphorylation of Smad2/3, induced ubiquitination of PPM1A and decreased the expression level of PPM1A, while TRIM47 RNA interference reversed this result. Significance Melittin has anti-fibrotic effect in HELF by directly reducing the phosphorylation of Smad2/3 or indirectly reducing the phosphorylation of Smad2/3 by decreasing the expression levels of TRIM47 whose overexpression induces ubiquitination of PPM1A.

It is meaningful for researching on monitoring and forecasting technology of slope stability to prevent landslide disasters, especially in the water system fields. Based on the mechanical principle of interaction among sliding mass, sliding bed and monitoring cable, a new type of energy absorbing cable (called Large Deformation Cable with Constant Resistance) which can have 2000mm deformation with constant pull load of 850kN is developed. The mechanical principle of relative movement between sliding mass and sliding bed is proposed, and the multi-factor monitoring is transformed into single landslide mechanical monitoring. The relationship between sliding force of slope and pretightening force of monitoring cable is set up. According to the physical model experiment of landslide, the sliding force at the potential slip surface will change continually before the landslide. When the sliding force is greater than the shear resistance at the potential slip surface, the landslide will take place, which means the variation of sliding force at the potential slip surface will be ahead of displacement on the ground of slope. Consequently, monitoring the variation of sliding force at the potential slip surface is better than that of the displacement. Based on above principle and experiment, the system of real-time remote monitoring and forecasting for landslide disasters based on the large deformation cable with constant resistance is developed, which can realize the real-time remote monitoring warning of sliding force. Several landslides have been successfully predicted.

Proteomic study on membrane-integrated proteins in endoplasmic reticulum (ER) fractions was performed. In this study, we examined the effects of heat stress on Jurkat cells. The ER fractions were highly purified by differential centrifugation with sodium carbonate washing and acetone methanol precipitations. The ER membrane proteins were separated by one dimensional electrophoresis (1-DE), and some of the protein bands changed their abundance by heat stress, 12 of the 14 bands containing 40 and 60 ribosomal proteins whose expression level were decreased, on the contrary, 2 of the 14 bands containing ubiquitin and eukaryotic translation initiation factor 3 were increased. Heat treatment of human Jurkat cells led to an increase in the phosphorylation of PERK and eIF2α within 30 min of exposure. This was followed by an increase in the expression of the GRP78. Protein ubiquitination and subsequent degradation by the proteasome are important mechanisms regulating cell cycle, growth and differentiation, the result showed that heat stress enhanced ubiquitination modification of the microsomal proteins. The data of this study strongly suggest that heat treatment led to a significant reduction in protein expression and activated UPR, concomitant with protein hyperubiqutination in ER.

Visible-light-initiated homogeneous oxidative synthesis of nitriles from amines was accomplished through a combined use of photoredox and copper catalysis. This transformation was performed at room temperature with O2 as the oxidant.

A new sorbent material, calcium fluoride-acid red 138 (AR138), was synthesized and characterized by various methods. The hybrid adsorbent for adsorption of PCBs corresponded to the octanol–water partition law, and their partition coefficients were calculated to be 5,836.8 mg kg−1 for PCB029, 8,311.9 mg kg−1 for PCB101, and 20,815 mg kg−1 for PCB180, respectively. When the hybrid adsorbent was used in the treatment of a polluted ground water sample, the removal of PCB concentrations was satisfactory. All the reactants used in the synthesis of the adsorbent material are easily available and are harmless to the environment. Also, the AR138 reactant may reuse a concentrated AR138-producing wastewater instead. Therefore, this work has developed a simple, eco-friendly, and “waste treat waste” method for the large-scale production of a cost-effective sorbent.

We performed proteomic differential display analysis of human malignant pleural mesothelioma (MPM) cell lines and a human pleural mesothelial cell line by using 2-DE and LC-MS/MS. The human MPM cell lines were NCI-H28, NCI-H2052 and NCI-H2452, and the human pleural mesothelial cell line was MeT-5A. Between MeT-5A and NCI-H2052, we found 38 protein spots whose expression levels were different, from the results of 2-DE; 28 protein spots appeared higher, and 10 other protein spots lower in NCI-H2052 than in MeT-5A. These spots were analyzed by LC-MS/MS analysis and identified by a peptide sequence tag. However, from the results of 2-DE of the other cell lines, there was only one consistently upregulated protein, astrocytic phosphoprotein PEA-15, in all three MPM cell lines. Western blotting using specific antibodies against PEA-15 confirmed the elevated expression level of PEA-15 in all three MPM cell lines compared with MeT-5A cells and normal pleura tissues from patients. PEA-15 was knocked down in NCI-H2052 cells, and the proliferation of PEA-15-silenced NCI-H2052 cells was suppressed 7-15% compared with negative control cells. These results suggest that PEA-15 expression is likely to be associated with the tumorigenesis of MPM.

Oral squamous cell carcinoma (OSCC) has an absolute majority of all oral cancer. We used proteomic technology to analyze the protein expression profile in OSCC tissues and accompanying surrounding normal tissues in four oral locations (buccal mucosa, gingival mucosa, oral floor, and tongue). Ten protein spots were overexpressed more strongly in cancer tissues than normal ones, and were identified as proliferating cell nuclear antigen, 14-3-3 ε, 14-3-3 σ, proteasome subunit α type 5, translationally controlled tumor protein, eukaryotic translation initiation factor 3 subunit, macrophage capping protein, and mitochondrial isocitrate dehydrogenase subunit α. Macrophage capping protein and mitochondrial isocitrate dehydrogenase subunit α had two spots. Especially, we focused on 14-3-3 σ protein, one of the eight identified proteins, and assessed its expression level in four oral locations of OSCC by using differential display methods. The expression level of 14-3-3 σ protein was upregulated in four locations of oral cavity. Eight proteins which we identified in this study may play an important role in OSCC carcinogenesis and progression and could be used as diagnostic biomarkers of OSCC.

There is a high burden of both diabetes and tuberculosis in China. Althoug both diabetes and tuberculosis have been the objects of intensive study the association between the two diseases is not fully understood. The aim of this study was to determine whether thetqualitative and quantitative differences in expression of some protein expression in plasma could be related to active pulmonary tuberculosis complicated with diabetes. Using two-dimensional gel electrophoresis and mass spectrometry we analyzed expression of the proteins associated with active pulmonary tuberculosis complicated with diabetes. Results showed that under the baseline condition, the levels of a-1 antitrypsin precursor, vitamin D-binding protein precursor, CD5 antigen like precursor, clusterin precursor, apolipoprotein A-I precursor, haptoglobin and fibrinogen g-chain differed between patients with active pulmonary tuberculosis and active pulmonary tuberculosis complicated with diabetes. Western blott results confirmed diffe...

Background There is a high burden of both diabetes and tuberculosis in China. Diabetes depresses the immunologic response that facilitates the development of infectious diseases, including infection by Mycobacterium tuberculosis, the agent of tuberculosis. Tuberculosis is the third cause of death among subjects with non-communicable diseases, and among the non-communicable diseases, diabetes is one of the most important. The relationship between diabetes and tuberculosis has already been object of many investigations but the association between these two diseases is not fully understood. The aim of this study was to determine whether relative qualitative and quantitative differences in protein expression of plasma could be related to active pulmonary tuberculosis complicated with diabetes. Methods Biological parameters are useful tools for understanding and monitoring complicated disease processes. Our study employed two-dimensional gel electrophoresis and mass spectrometry to analyze the proteins associated with active pulmonary tuberculosis complicated with diabetes. Results Under the baseline condition, we found that the levels of α-1 antitrypsin precursor, vitamin D-binding protein precursor, CD5 antigen like precursor, clusterin precursor, apolipoprotein A-I precursor, haptoglobin, and fibrinogen γ-chain differed between patients with active pulmonary tuberculosis and active pulmonary tuberculosis complicated with diabetes subjects. Western blotting results confirmed differential expression of clusterin. Conclusions We identified active pulmonary tuberculosis complicated with diabetes-associated proteins in plasma. C-terminal haptoglobin is a possible candidate protein of interest, which might be a link between active pulmonary tuberculosis and diabetes. The dynamics of protein expression during disease progression may improve our understanding of the pathogenesis of active pulmonary tuberculosis complicated with diabetes.

Two-dimensional gel electrophoresis (2-DE) and tandem mass spectrometry were successfully used for determination of a phosphorylation site of stathmin induced by heat stress to Jurkat cells of a human T lymphoblastic cell line. The cells were incubated for 30 min at 41 degrees C up to 45 degrees C in a serum free 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffered culture medium. The intracellular soluble proteins were separated by 2-DE, and some of the proteins increased their abundance by heat stress. Those proteins were identified to be calmodulin, protein kinase C substrate, thymosin beta-4 and F-actin capping protein beta-subunit by peptide mass fingerprinting (PMF) with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). On the contrary, protein phosphatase 2C gamma-isoform, nucleophosmin, translationally controlled tumor protein, Rho GDP-dissociation inhibitor-1, eukaryotic translation initiation factors 5A and 3A subunit 2, ubiquitin-like protein SMT 3B and chloride intracellular channel protein-1 were decreased their abundance. A protein spot of M(r) 18,000 and pI 5.9 was markedly increased at temperatures higher than 43 degrees C at which the cells were led to apoptosis. The spot was identified to be stathmin of a signal relay protein which has a function of sequestering microtubule. MALDI-quadrupole ion trap (QIT)-TOF-MS/MS and immunoblotting with a monoclonal antibody specific for a phosphorylation site of stathmin showed that the spot was a phosphorylated stathmin at serine 37 (Ser 37). The phosphorylation was suppressed by treatment of cells with olomoucine of an inhibitor specific for cyclin dependent kinase (Cdk-1). These results strongly suggest that heat stress activates Cdk-1 which phosphorylates Ser 37 on the stathmin molecule. The phosphorylation may cause the functional loss of stathmin for dynamic microtubule assembly and leads Jurkat cells to cell cycle arrest and apoptosis.

In order to effectively employ the unique high temperature mechanical properties of carbon/carbon composite substrates, SiC coatings reinforced by SiC whiskers were prepared by pack cementation method. The effect of SiC whiskers on the oxidation resistance properties of the single-layer coating and double-layer coating was investigated. SiC whiskers in the single-layer SiC coating have little effect on the anti-oxidation property but obviously improve the thermal shock property. The double-layer coating with inner-layer reinforced coating exhibits more perfect anti-oxidation ability than the double-layer coating with SiC inner-layer coating.

Studies on blood plasma proteome were performed by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) using samples supplied from China and the United States by collaborations of Human Plasma Proteome Project of HUPO. The plasma proteins were separated by isoelectric focusing using capillary type polyacrylamide gel (PAG) with carrier ampholytes pH 3.5-10 in the first dimension and with SDS-PAGE using 15%, 12.5% and 8% gel in the second dimension. The protein spots were identified by peptide mass fingerprinting using electrospray ionization MS/MS (LC/MSD Trap XCT, Agilent). More than 145 protein spots were identified by LC-MS/MS. The 2-DE patterns showed that the samples stored at 4°C for 3 days lost inter-alpha-trypsin inhibitor heavy chain 4 (ITIH-4).

Serum antibodies against pathogenic bacteria play immunologically protective roles, and can be utilized as diagnostic markers of infection. This study focused on Japanese child serum antibodies against Helicobacter pylori, a chronically-infected gastric bacterium which causes gastric cancer in adults. Serological diagnosis for H. pylori infection is well established for adults, but it needs to be improved for children. Serum samples from 24 children, 22 H. pylori (Hp)-positive and 2 Hp-negative children, were used to catalogue antigenic proteins of a Japanese strain CPY2052 by two-dimensional electrophoresis followed by immunoblot and LC-MS/MS analysis. In total, 24 proteins were identified as candidate antigen proteins. Among these, the major virulence factor, cytotoxin-associated gene A protein (CagA) was the most reactive antigen recognized by all the Hp-positive sera even from children under the age of 3 years. The major antigenic part of CagA was identified in the middle region, and two peptides containing CagA epitopes were identified using a newly developed peptide/protein-combined array chip method, modified from our previous protein chip method. Each of the epitopes was found to contain amino acid residue(s) unique to East Asian CagA. Epitope analysis of CagA indicated importance of the regional CagA antigens for serodiagnosis of H. pylori infection in children.

The purpose of this study was to analysis on the affection of key protein expression by vitamin D3, and to investigate the mechanism of vitamin D3 on contributing to diabetes.Good status U937 cells were cultured by high glucose medium which contained 45 mmol/L D-Glucose, and treated by 1, 25-(OH)(2)-D3. The control groups were U937 cells cultured by high glucose. The whole cell proteins were extracted and separated by 2-dimentional gel electrophoresis (2-DE), differential expression proteins were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS).There were 30 protein spots whose expression was significantly different between vitamin D-treated and untreated high glucose loaded cells. Six protein spots were identified by MALDI-TOF-MS, three of which, prohibitin, heat shock 70 kD protein 8, and catalase were protection of cell stress proteins, one of which, electron transport flavoprotein, was respiratory chain related proteins, two of which, fumarylacetoacetate hydrolase and triosephosphate isomerase, were cell metabolism related proteins.The differentially expressed proteins of high glucose load human U937 cell line stimulated by vitamin D3 were found. These differential proteins were mainly related to oxidative stress and energy metabolism. These data suggested that 1, 25-(OH)(2)-D3 might regulate cell high glucose load by reduction oxidative stress injury and affection energy metabolism.

A new kind of acid red 138 (AR138)–calcium fluoride nanosized CaF2/AR138 hybrid adsorbent (CFAR) was prepared. The microstructure of the hybrid adsorbent was characterized and its adsorption capacity to two cationic dyes, methyl blue (MB) and ethyl violet (EV), from aqueous solutions was examined using a batch sorption technique. The adsorption behavior of the MB and EV on CFAR is in good agreement with the Langmiur isotherms. The maximum adsorption capacity (Qm) of CFAR for MB and EV are 370.4 and 131.6 mg/g, respectively. The effects of pH, ionic strength, temperature, and time have been investigated in detail. The CFAR adsorbent was used to treat two dye wastewaters with satisfactory results. In synthesis of the materials, all of the reactants are easily available and harmless to environment as well because AR138 may use a concentrated AR138-producing wastewater instead. Therefore, this paper has developed a simple, eco-friendly and waste treat waste method for large-scale production of a cost...

Pancreatic cancer (PC) is characterized by rapid tumor spread, and very few patients with PC survive for more than 5 years. It is imperative to discover additional diagnostic biomarkers or specific therapeutic targets in order to improve the treatment of patients with PC. In search for useful biomarkers, we analyzed ten pairs of non-cancerous and cancer tissues from patients with PC by two-dimensional gel electrophoresis (2-DE). Nineteen protein spots showed differential expression on 2-DE gels between the cancer and non-cancerous tissues. Six upregulated protein spots were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) as calreticulin, glutathione synthetase, stathmin, vinculin, α-enolase and glyceraldehyde-3-phosphate dehydrogenase. Western blotting demonstrated that vinculin was predominantly expressed in the pancreatic cancer tissues compared with to non-cancerous tissues. Our findings indicate that vinculin may be a clinically useful biomarker of PC.

Pancreatic cancer (PC) is one of the most deadly malignant tumors. The aim of this study was to identify potential biomarkers for PC. Using two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry, the proteomic profiles of pancreatic cancerous and non-cancerous tissues from ten patients with PC were compared. One of the numerous spots that showed stronger intensity in cancerous compared to non-cancerous tissues was identified as non-muscle cofilin (cofilin-1). This up-regulation was validated by Western blot analysis. It is noteworthy that Western blot analysis showed significantly lower expression of muscle cofilin (cofilin-2) in pancreatic cancerous tissues compared to non-cancerous tissues. This is the first time that cofilin isoforms (cofilin-1/2) have been identified to be differentially expressed in pancreatic cancerous tissues. Therefore, cofilin isoforms may serve as candidates for clinically useful biomarkers or therapeutic targets for PC.

TS-1 is an oral anticancer agent containing two biochemical modulators for 5-fluorouracil (5-FU) and tegafur (FT), a metabolically activated prodrug of 5-FU. TS-1 has been recognized as an effective anticancer drug using standard therapies for patients with advanced pancreatic cancer along with gemcitabine. However, a high level of inherent and acquired tumor resistance to TS-1 induces difficulty in the treatment. To identify proteins linked to the TS-1-resistance of pancreatic cancer, we profiled protein expression levels in samples of TS-1-resistant and -sensitive pancreatic cancer cell lines by using two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The cytotoxicity of a 5-FU/5-chloro-2,4-dihydroxypyridine (CDHP) combination towards pancreatic cancer cell lines was evaluated by MTS assay. Panc-1, BxPC-3, MiaPaCa-2 and PK59 showed high sensitivity to the 5-FU/CDHP combination (TS-1-sensitive), whereas PK45p and KLM-1 were much less sensitive (TS-1-resistant). Proteomic analysis showed that eleven spots, including T-complex protein 1 subunit beta, ribonuclease inhibitor, elongation factor 1-delta, peroxiredoxin-2 and superoxide dismutase (Cu-Zn), appeared to be down-regulated, and 29 spots, including hypoxia up-regulated protein 1, lamin-A/C, endoplasmin, fascin and annexin A1, appeared to be up-regulated in TS-1-resistant cells compared with -sensitive cells. These results suggest that the identified proteins showing different expression between TS-1-sensitive and -resistant pancreatic cancer cells possibly relate to TS-1-sensitivity. These findings could be useful to overcome the TS-1-resistance of pancreatic cancer cells.

Our earlier studies reported that ONO-4007, a synthetic lipid A analogue with low endotoxic activity, had shown much effect on tumour necrosis factor (TNF)-alpha-sensitive rat hepatoma cells, even though it had no effect on TNF-alpha-resistant cells.To find biomarkers which relate to the sensitivity of cancer cells to TNF-alpha, proteomic differential display analysis for TNF-alpha-resistant cKDH-8/11 and -sensitive KDH-8/YK rat hepatoma cell lines was carried out using two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry.Two-DE analysis showed 32 spots, whose expression was different between cKDH-8/11 cells and KDH-8/YK cells. Of these, 12 were up-regulated and 20 were down-regulated in cKDH-8/11 cells compared to KDH-8/YK cells. The up-regulated proteins include transitional endoplasmic reticulum ATPase, 78 kDa glucose-regulated protein (GRP78), heat-shock cognate 71 kDa protein (HSC71) and protein disulfide-isomerase A6. The down-regulated proteins included alpha-enolase, aldose reductase, glutathione reductase, annexin A1, glutamate dehydrogenase 1 and dihydrolipoyl dehydrogenase.These findings suggest that these differentially regulated proteins could be factors responsible for the resistance of cKDH-8/11 cells to TNF-alpha-induced cell death.

The understanding of tumour progression is one of the most important strategies to conquer tumour. QR-32 is a regressive murine fibrosarcoma cell line, and QRsP-11 is a progressive malignant tumour cell clone derived from QR-32. Ina recently published study a differential display analysis for the cytoplasmic proteins was shown by using two-dimensional gel electrophoresis (2-DE) making full use of isoelectric focusing capillary gels and Coomassie brilliant blue R-250 staining. Furthermore, a differential display analysis of the nuclear proteome for QR-32 and QRsP-11 was performed. The present study shows a non-nuclear proteomic differential display analysis, using 2-DE making full use of immobilized pH gradient strips and Flamingo™ fluorescent gel stain, between QR-32 and QRsP-11 to identify particular proteins which may be involved in malignant progression. In QRsP-11 25 proteins were up-regulated, including hypoxia up-regulated protein 1, and 6 were down-regulated compared with QR-32. These results suggest that the identified non-nuclear proteins showing different expression between QR-32 and QRsP-11 possibly related to malignant tumour progression.

Pancreatic cancer (PC) is one of the most lethal malignant tumors because of late diagnosis and the lack of response to various therapies. To identify potential biomarkers in cancerous serum from early stage PC patients, we carried out two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to compare the serum proteomic profiles from 45 patients with PC and 20 healthy volunteers. Seven spots showed differential expression on 2-DE gels and two up-regulated protein spots were identified by LC-MS/MS as α-1-antitrypsin (AAT). These protein spots were also confirmed by Western blotting. This is the first time that AAT isoforms have been identified as potential serum biomarkers for PC. The serum isoforms of AAT may be clinically useful for PC diagnosis and monitoring.

This study performed proteomic differential display analysis of human scirrhous-type gastric carcinoma (SGC) cell lines and normal gastric mucosa (NGM) tissues by using two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The human SGC cell lines were OCUM-1, OCUM-2M, OCUM-2MLN, OCUM-2D, OCUM-D3, OCUM-9 and OCUM-12. Among the SGC cell lines and the NGM tissues, 28 protein spots were found whose expression levels were different from the results of 2-DE: 19 protein spots appeared higher, and 9 other protein spots appeared lower in SGCs than in NGM tissues. These spots were analysed by LC-MS/MS analysis and identified by a peptide sequence tag. Identified increased spots included elongation factor 1-beta, 14-3-3 sigma, tropomyosin alpha-4 chain, protein DJ-1, nucleoside diphosphate kinase A, elongation factor Tu and peroxiredoxin-1. Western blot analysis showed increased protein level of 14-3-3 sigma in SGCs. Although OCUM-1 and AGS (gastric cancer) showed up-regulation of 14-3-3 sigma, MiaPaca-2 (pancreatic cancer), Huh-7 (HCC) and NCI-H2052 (malignant pleural mesothelioma) showed very weak expression of 14-3-3 sigma. The up-regulation of 14-3-3 sigma may play an important role in SGC carcinogenesis and progression and may be used as a diagnostic biomarker of SGC.

Highly sensitive Coomassie brilliant blue SeePico™ Stain was applied for proteomic analysis using two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). After staining with Flamingo™ Fluorescent Gel Stain, the images of the protein spots were analyzed, and 424 protein spots were detected. After washing with Milli-Q water three times, the gels were re-stained with SeePico™ Stain and the images of the protein spots were analyzed; 272 spots were detected. To assess whether SeePico™ Stain alters MS analysis, a spot was picked up and was analyzed by LC-MS/MS. The MS analysis showed good protein identification. These results show a possible role for SeePico™ Stain in cancer proteomics using 2-DE and MS.