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The gut microbiota has emerged as an important contributor to the immune system and it is believed to be regulated by dietary fiber. In this study, therefore, the immunomodulatory and gut microbiota modulatory effects of polysaccharides from bee collected pollen of Chinese wolfberry (WBPPS) in cyclophosphamide (CTX)-treated mice were investigated. The results showed that WBPPS could enhance thymus and spleen indices and promote production of immune-related cytokines (IgA, TNF-α, TGF-β1 and IL-10) in serum of CTX-treated mice. In addition, WBPPS could significantly improve hepatic function and oxidative stress in CTX-treated mice via increasing the levels of GSH, SOD and CAT and decreasing the levels of ALT and MDA. More importantly, WBPPS remarkably increased the relative abundances of Family_XIII, Ruminococcaceae and Saccharimonadaceae while decreased the relative abundances of Tannerellaceae, Marinifilaceae and Rikenellaceae, which were closely associated with immune traits. Overall, WBPPS might be developed as a promising immunomodulator in functional foods.

The ventromedial hypothalamus (VMH) is a primary site that initiates counter-regulatory response and regulates food intake, maintaining glucose and lipid homeostasis. Known as a nutrient sensor and metabolic regulator, O-linked β-D-N-acetylglucosamine modification (O-GlcNAcylation) of intracellular proteins is catalyzed by O-GlcNAc transferase (OGT). How O-GlcNAc signaling in the VMH regulates whole-body metabolism remains unknown. Here, using the mouse model with VMH neuron-specific OGT deletion (VOK), we report that OGT is a nutrient sensor in VMH neurons that primarily regulates lipid metabolism. VOK mice show increased body weight and adiposity as well as reduced energy expenditure. OGT deletion in VMH neurons has no significant effects on food intake, physical activity, substrate utilization, glucose tolerance or insulin sensitivity. Lipolysis in subcutaneous and visceral white adipose tissues (WATs) is reduced in VOK mice, indicated by reduced activation of hormone-sensitive lipase (HSL) via cAMP-dependent protein kinase (PKA). In addition, VOK mice have lower serum glucagon and norepinephrine levels and show impaired depot-specific sympathetic innervation to WATs. These data reveal that O-GlcNAc signaling in VMH neurons specifically controls lipid metabolism through depot-specific sympathetic innervation. Disclosure Q. Wang: None. B. Zhang: None. Y. Yang: None. J. Mi: None. G. Tian: None. Q. Ong: None. X. Yang: None. Funding American Diabetes Association (1-19-IBS-119 to X.Y.); National Institutes of Health (R01DK089098, R01DK102648)

Lycium barbarum L. (abbreviated to L. barbarum) is a traditional medicinal and edible plant cultivated in different ecological areas in China. In this study, “Ningqi” No.1, one of the main cultivars of L. barbarum, was collected from 10 different important cultivation areas in 5 autonomous regions or provinces of China. Partial least squares regression and correlation analysis were used to analyze the relationship between 16 ecological factors, topographical, meteorological and soil factors and 9 phytochemicals of “Ningqi” No.1 fruits. There was no significant difference in carotenoids or ash in plants from the 5 regions or provinces (P ≥ 0.05), but total sugar content and total reducing sugars were lowest in Ningxia (P

Skeletal muscle preservation is a dynamic process that involves constant repair and regeneration. However, the regenerative capacity of muscle cells declines in hyperglycemia. This study aimed to explore the molecular mechanisms underlying this glucotoxicity during myoblast differentiation. C2C12 cells were exposed to different concentrations of glucose, to recapitulate the development of skeletal muscles in vivo in normo- and hyperglycemic conditions. In high glucose conditions, we found significant increases in levels of total cellular reactive oxygen species (ROS) and a reorganization of SUMO enzyme transcripts and SUMOylated proteins. Furthermore, in anticipation of the ROS-induced damage to newly formed myotubes, we observed acceleration of myogenesis. Interestingly, we found a tight relationship between SUMOylation of the Histone methyltransferase SET7/9 and the maintenance of sarcomeric structures of newly formed myotubes. Finally, treatment with the antioxidant anacardic acid preserved the function and activity of myotubes generated in high-glucose conditions by interfering with both ROS and SUMO pathways. Combined, these results suggest that increased oxidative stress and modulation of SUMO reactions are key mediators of glucotoxicity and inhibition of these perturbations using antioxidants might improve muscle regeneration in hyperglycemia.

Singlet oxygen is regarded as the primary cytotoxic agent in cancer photodynamic therapy (PDT). Despite the advances in optical methods to image singlet oxygen, it remains a challenge for in vivo application due to the limited tissue penetration depth of light. Up to date, no singlet oxygen-specific magnetic resonance imaging (MRI) probe has been reported. Herein, a T2 -weighted MRI probe is reported to visually detect singlet oxygen generated in PDT in vitro and in vivo. The MRI probe Ce6/Fe3 O4 -M is constructed by co-encapsulation of photosensitizer Ce6 and Fe3 O4 nanoparticles in mPEG2000 -TK-C16 micelles. Thioketal (TK) linker in the probe is highly sensitive to singlet oxygen, but lowly sensitive to other reactive oxygen species (ROS) existing in physiological and pathological environments. Singlet oxygen, generated with light irradiation, triggers the cleavage of TK, which leads to loss of surface polyethylene glycol, increment of the hydrophobicity, and aggregation of Fe3 O4 nanoparticles. Subsequently, negatively enhanced T2 -weighted MRI signal is obtained for visual detection of singlet oxygen in the solution, cancer cells, and in vivo. This oxidation responsive MRI probe is expected to hold great promise in evaluating the ability of photosensitizers to generate singlet oxygen and in predicting the therapeutic efficacies of PDT in vivo.

Neoadjuvant chemotherapy (NCT) is a standard care for esophageal squamous cell carcinoma (ESCC), but the efficacy is unsatisfactory. Cancer stem cells (CSCs) play key roles in chemotherapy resistance. Gene amplified in squamous cell carcinoma 1 (GASC1) is a neoteric gene in stemness maintaining of ESCC. We aimed to reveal whether GASC1 could be a predictive biomarker for NCT in ESCC. ESCC patients (T2-4N0-2M0) were evaluated for GASC1 expression using immunohistochemical staining and classified as GASC1-low group (GLG) and GASC1-high group (GHG). NCT was delivered in two cycles and then the surgery was completed. Primary endpoints were tumor regression grade (TRG) and objective response rate (ORR); secondary endpoints were radical surgical resection (R0) rate and three-year overall survival (OS). 60 patients were eligible with evaluable outcomes: 24 in GHG and 36 in GLG. Between GHG and GLG, TRG1, TRG2, TRG3, and TRG4 were 0 : 16.7%, 20.8% : 41.7%, 58.3% : 36.1%, and 20.8% : 5.6%, respectively (P=0.006); ORR and R0 rate were 33.3% : 69.4% (P=0.006) and 75% : 94.4% (P=0.046), respectively; the median OS was 20 : 32 (months) (P=0.0356). No significant difference in the three-year OS was observed between GHG and GLG: 29.2% : 41.7% (P=0.24). Furthermore, the GASC1 expression level was associated with poor OS independent of other factors by univariate and multivariate analyses. Therefore, GASC1 might be a potential biomarker to predict NCT efficacy for ESCC.

Fitness is closely associated with the development of pesticide resistance in insects, which determines the control strategies employed to target species and the risks of toxicity faced by non-target species. After years of selections with beta-cypermethrin in laboratory, a strain of housefly was developed that was 684,521.62-fold resistant (CRR) compared with the susceptible strain (CSS). By constructing ≤ 21 d and ≤ 30 d life tables, the differences in life history parameters between CSS and CRR were analyzed. The total production numbers of all the detected development stages in CRR were lower than in CSS. Except for the lower mortality of larvae, all the other detected mortalities in CRR were higher than in CSS. ♀:♂ and normal females of CRR were also lower than those of CSS. For CRR, the relative fitness was 0.25 in the ≤ 21 d life table and 0.24 in the ≤ 30 d life table, and a lower intrinsic rate of increase (rm) and net reproductive rate (Ro) were detected. Based on phenotype correlation and structural equation model (SEM) analyses, fecundity and viability were the only directly positive fitness components affecting fitness in CRR and CSS, and the other components played indirect roles in fitness. The variations of the relationships among fitness, fecundity and viability seemed to be the core issue resulting in fitness differences between CRR and CSS. The interactions among all the detected fitness components and the mating frequency-time curves appeared to be distinctly different between CRR and CSS. In summary, fecundity and its related factors separately played direct and indirect roles in the fitness costs of a highly beta-cypermethrin-resistant housefly strain.

Non‐alcoholic fatty liver disease (NAFLD) is one of the most common causes of hepatocellular carcinoma (HCC), but the underlying mechanisms behind the correlation of NAFLD with HCC are unclear. We aimed to uncover the genes and potential mechanisms that drive this progression. This study uncovered the genes and potential mechanisms through a multiple ’omics integration approach. Quantitative proteomics combined with phenotype‐association analysis was performed. To investigate the potential mechanisms, a comprehensive transcriptome/lipidome/phenome‐wide association analysis was performed in genetic reference panel BXD mice strains. The quantitative proteomics combined with phenotype‐association results showed that VDAC1 was significantly increased in tumor tissues and correlated with NAFLD‐related traits. Gene co‐expression network analysis indicated that VDAC1 is involved in mitochondria dysfunction in the tumorigenic/tumor progression. The association between VDAC1 and mitochondria dysfunction can be explained by the fact that VDAC1 was associated with mitochondria membrane lipids cardiolipin (CL) composition shift. VDAC1 was correlated with the suppression of mature specie CL(LLLL) and elevation level of nascent CL species. Such profiling shift was supported by the significant positive correlation between VDAC1 and PTPMT1, as well as negative correlation with CL remodeling enzyme Tafazzin (TAZ). This study confirmed that the expression of VADC1 was dysregulated in NAFLD‐driven HCC and associated with NAFLD progression. The VDAC1‐driven mitochondria dysfunction is associated with cardiolipin composition shift, which causes alteration of mitochondria membrane properties.

Among patients with the rare diagnosis of pancreatic neuroendocrine tumor (P-NET), a substantial proportion suffer from the inherited cancer syndrome multiple endocrine neoplasia type 1 (MEN1), which is caused by germline mutations of the MEN1 suppressor gene. Somatic mutations and loss of the MEN1 protein (menin) are frequently also found in sporadic P-NETs. Thus, a human neuroendocrine pancreatic cell line with biallelic inactivation of MEN1 might be of value for studying tumorigenesis. We used the polyclonal human P-NET cell line BON1, which expresses menin, serotonin, chromogranin A and neurotensin, to generate a monoclonal stable MEN1 knockout BON1 cell line (MEN1-KO-BON1) by CRISPR/Cas9 editing. Changes in morphology, hormone secretion, and proliferation were analyzed, and proteomics were assessed using nanoLC-MS/MS and Ingenuity Pathway Analysis (IPA). The menin-lacking MEN1-KO-BON1 cells had increased chromogranin A production and were smaller, more homogenous, rounder and grew faster than their control counterparts. Proteomic analysis revealed 457 significantly altered proteins, and IPA identified biological functions related to cancer, e.g., posttranslational modification and cell death/survival. Among 39 proteins with at least a two-fold difference in expression, twelve are relevant in glucose homeostasis and insulin resistance. The stable monoclonal MEN1-KO-BON1 cell line was found to have preserved neuroendocrine differentiation, increased proliferation, and an altered protein profile.

Transgenic adenocarcinoma of the mouse prostate (TRAMP) mice is a widely used transgenic animal model of prostate cancer (PCa). We performed a label-free quantitative proteomics analysis combined with a bioinformatics analysis on the entire prostate protein extraction from TRAMP mice and compared it with WT littermates. From 2379 total identified proteins, we presented a modest mice prostate reference proteome containing 919 proteins. 61 proteins presented a significant expression difference between two groups. The integrative bioinformatics analysis predicted the overexpression of platelet-derived growth factor B (PDGF-B) in tumor tissues and supports the hypothesis of the PDGF-B signaling network as a key upstream regulator in PCa progression. Furthermore, we demonstrated that Crenolanib, a novel PDGF receptor inhibitor, inhibited PCa cell proliferation in a dose-dependent manner. Finally, we revealed the importance of PDGF-B regulatory network in PCa progression, which will assist us in understanding the role and mechanisms of PDGF-B in promoting cancer growth and provide valuable knowledge for future research on anti-PDGF therapy.

Plasmonic metal nanostructures possessing plentiful gaps or tips have attracted great attention in surface enhanced Raman scattering (SERS) due to their superior performance. Herein, we present an efficient fabrication strategy to construct a large-area and highly ordered 3D silver “urchin-like” array (SULA) based on the ultra-thin alumina membrane (UTAM) mask and the self-assembled polystyrene spheres template. Numerical simulations reveal that the proposed SULA possesses enormous hotspots at the tips of Ag nanocones and in the crevice gaps between the adjacent Ag nanocaps responsible for high sensitivity with an enhancement factor of 1.8 × 106 and excellent reproducibility with a relative standard deviation of 8.3% in SERS analysis. Furthermore, the SULA substrate exhibits great SERS performance for detecting the contaminant crystal violet and thiram in real water environments and excellent linear intensity-concentration dependence relationships. The results indicate the proposed SULAs are promising to be developed as high-performance SERS platforms in qualitative and quantitative trace contaminants in complex water environments.

BACKGROUND Diabetes has become a global public health problem and danger to human health. Diabetes is the main cause of blindness, kidney failure, heart attack, stroke, and lower limb amputation. According to the latest epidemiological survey and research, the overall prevalence of diabetes in mainland China is 11.2%, of which type 2 diabetes mellitus (T2DM) accounts for more than 90% acupuncture combined with Chinese herbal medicine have been widely used in the treatment of T2DM. However, we have not found a meta-analysis of their synergistic effects. Therefore, this systematic review and meta-analysis aims to evaluate the efficacy and safety of acupuncture combined with Chinese herbal medicine in the treatment of T2DM. METHOD From inception up to September 20, 2021, the PubMed, Web of Science, Embase, AMED, Cochrane Library, CNKI, VIP, CBM, and Wanfang databases will be searched. The publication date or language will not be limited. We will apply a combination of medical keywords, including "acupuncture", "Chinese herbal medicine", and "type 2 diabetes mellitus". We will also check other ongoing and unpublished studies in the clinical trial registry. At the same time, we will manually search all reference lists from relevant systematic reviews to find other eligible studies. We will use Review Manager software (REVMAN v5.3 Cochrane Collaboration) to meta-analyze the selected literature. The study for acupuncture combined with Chinese herbal medicine in the treatment of T2DM was a randomized controlled study. Two researchers will independently review the research selection, data extraction, and research quality assessments. Finally, we will observe the outcome measures. RESULTS This study will generate evidence-based data on the treatment of T2DM with acupuncture combined with Chinese herbal medicine and will provide new ideas and treatment modalities to investigate in future research.

Ti3+ self-doped mesoporous black TiO2/graphene assemblies are fabricated by a facile solvothermal method and surface hydrogenation. The structure, crystallinity, morphology, and chemical state of the as-prepared samples are characterized in detail by X-ray diffraction, Raman, X-ray photoelectron spectroscopy, transmission electron microscopy, N2 adsorption and UV-visible diffuse reflectance spectroscopy. The results show that the presence of Ti3+ can efficiently extend the photoresponse of anatase TiO2 to visible light region. The solar-driven photocatalytic hydrogen evolution shows that Ti3+ self-doped mesoporous black TiO2/graphene assemblies exhibit the highest photocatalytic activity (186μmolh-1 0.01g-1), exceeding to mesoporous TiO2/graphene assemblies and mesoporous black TiO2. It also exhibits superior photoelectrochemical properties compared with mesoporous TiO2/graphene assemblies. The unpredicted-high photocatalytic performance is attributed to the close contact between the unique two-dimensional graphene structures coupled with TiO2 mesoporous architectures resulting in outstanding charge separation efficient and the Ti3+ self-doping extending the utilization ratio of visible light.

This paper proposes a new conceptual configuration for oscillating surge wave energy converters. The concept is a self-floating device consisting of two vertical flaps with a common hinge and an averaged density equal to that of water. The two flaps will oscillate in opposite directions when driven by incident waves. Kinetic energy will be extracted from the relative speed between the two flaps by a power take-off embedded at the hinge. Numerical studies reveal that the self-floating device has a smaller resonant period and optimal power take-off damping compared with a bottom-hinged floating device of the same overall dimensions. As long as the power take-off damping is appropriately tuned, the opposite-rotation mechanism is valid in both regular and irregular waves, and no matter whether the hinge is fixed or connected with mooring lines. With the self-floating feature and opposite-rotation mechanism, the proposed concept can potentially benefit deep ocean applications by exerting a significantly less reaction loading on mooring lines and eliminating the need for a supporting frame to house the power take-off. The influence of various design parameters such as flap dimensions, submerged depth, incident wave direction, and water depth is also analyzed for a comprehensive evaluation of the proposed concept.

Current challenge of using cytokines is its poor distribution and systemic side effects. To avoid this issue, we prepared the tumor-targeted and microenvironment-responsive nanocarriers (TRN), which were consisted of α-tocopheryl succinate (α-TOS) loaded mesoporous silica nanoparticles as cores, and surface-modified by thioketal-linkage, electrostatically coated with carboxymethyl chitin, and further anchored glucose-regulated protein 78-binding peptide as shells for encapsulating IL-12. TRN showed a size of 260 nm after encapsulated IL-12 and α-TOS with loading content of 0.0206% and 7.21%, respectively, and exhibited good biocompatibility to 4 T1 cells and macrophages. Moreover, IL-12/α-TOS loaded TRN displayed obvious anti-tumor efficacy on BALB/c nude mice bearing 4 T1 tumors, which was derived from promoted targeting to tumor tissue, endocytosed by macrophages and locally release IL-12 to subsequently repolarize tumor-associated macrophages into tumoricidal M1 phenotype with reduced side effects. The nanosystem exhibited as a promising strategy with functional conversion of macrophages in tumor microenvironment for anti-tumor therapy.

Nanotechnology provides a powerful tool to overcome many disadvantages of small-molecule photosensitizers for photodynamic cancer therapy, such as hydrophobicity, rapid blood clearance, low accumulation in tumor tissue and low cell penetration, etc. The occurrence of quench in photosensitizer-loaded nanoparticle greatly downregulates the ability to generate singlet oxygen with light irradiation. Stimuli-responsive nanocarriers can improve the efficacy of PDT to a certain extent. However, insufficient release of photosensitizer from either endogenous or exogenous stimuli responsive nanocarriers in the short period of light irradiation restricts full usage of the photosensitizer delivered into cancer cells. We here report a dual-step light irradiation strategy to enhance the efficacy of cancer PDT. Ce6 as a photosensitizer is loaded in singlet oxygen-sensitive micelles (Ce6-M) via self-assembly of amphiphilic polymer mPEG2000-TK-C16. After co-incubation of Ce6-M with cancer cells or i.v. injection of Ce6-M, cancer cells or tumor tissues are irradiated with light for a short time to trigger Ce6 release, and 2 h later, re-irradiated for relatively long time. The sufficient release of Ce6 in the period between twice light irradiation significantly improves the generation of singlet oxygen, leading to more efficient cancer therapeutic effects of dual-step irradiation than that of single-step irradiation for the same total irradiation time.

The most commonly used vibration energy harvesting system is the single-degree-of-freedom (SDOF) system. However, the power output of the SDOF system is limited by the mechanical damping ratio and the efficiency is only 50% at the maximal power output. In this paper, we modified the SDOF configuration by adding a spring in series with the energy transducer, which results in a configuration where a parallel energy-harvesting damper and inerter are in series with the spring, similar to the Maxwell model. The parameters of the proposed configuration are analyzed and optimized. The optimization results show the new configuration can produce many times more power output compared with the conventional SDOF system if the primary spring is relatively soft and the added spring is properly selected. In addition, the energy-harvesting efficiency can be significantly increased at the maximal power output. A lab experiment of backpack setup demonstrated 82% power increase and an efficiency of 83%. Lab test using a bridge vibration induced by freight train finds out that the proposed configuration can double the power output compared with the conventional SDOF system. With the augment of a simple spring, the proposed configuration disrupts the fundamental limit on the power and efficiency of the conventional SDOF energy-harvesting system, which opens a new perspective in the energy-harvesting field.

The development of road traffic communication technology makes communication means more diverse. With the increase of the complexity of urban road traffic communication network, the fault diagnosis of road traffic communication network has been paid more and more attention. The alarm data in the communication network is the manifestation of network failure. Network operation and maintenance personnel can understand the current network operation status according to the alarm information displayed in the system, and then analyze, locate and handle the fault. Therefore, the identification of alarm information is very important for the timely treatment of network failure. In this paper, based on the existing communication network theory and network equipment alarm information, this paper proposes a method using SVM, which can effectively solve the fault identification of network communication input anomaly.

Ocular neovascularization is a devastating pathology of numerous ocular diseases and is a major cause of blindness. Caveolin-1 (Cav-1) plays important roles in the vascular system. However, little is known regarding its function and mechanisms in ocular neovascularization. Here, using comprehensive model systems and a cell permeable peptide of Cav-1, cavtratin, we show that Cav-1 is a critical player in ocular neovascularization. The genetic deletion of Cav-1 exacerbated and cavtratin administration inhibited choroidal and retinal neovascularization. Importantly, combined administration of cavtratin and anti-VEGF-A inhibited neovascularization more effectively than monotherapy, suggesting the existence of other pathways inhibited by cavtratin in addition to VEGF-A. Indeed, we found that cavtratin suppressed multiple critical components of pathological angiogenesis, including inflammation, permeability, PDGF-B and endothelial nitric oxide synthase expression (eNOS). Mechanistically, we show that cavtratin inhibits CNV and the survival and migration of microglia and macrophages via JNK. Together, our data demonstrate the unique advantages of cavtratin in antiangiogenic therapy to treat neovascular diseases.

// Tingting Men 1, * , Cuicui Yu 2, * , Dan Wang 3 , Fang Liu 3 , Jingjing Li 3 , Xiaoying Qi 3 , Chunhua Yang 3 , Wenguo Jiang 3 , Xiaodan Wei 3 , Xuri Li 3 , Bin Wang 4 , Jia Mi 3 and Geng Tian 3 1 School of Nursing, Binzhou Medical University, Yantai, Shandong, China 2 Department of Anesthesiology, Yantai Yu Huang Ding Hospital, Yantai, Shandong, China 3 Medicine and Pharmacy Research Center, Binzhou Medical University, Yantai, Shandong, China 4 Institute of Molecular Imaging, Binzhou Medical University, Yantai, Shandong, China * These authors have contributed equally to this work Correspondence to: Geng Tian, email: tiangengshandong@yeah.net Jia Mi, email: Jia.mi@kemi.uu.se Bin Wang, email: binwang001@aliyun.com Keywords: prostate cancer, interleukin-10, polymorphism, peripheral blood interleukin-10, meta-analysis Received: December 16, 2016 Accepted: April 11, 2017 Published: April 29, 2017 ABSTRACT This study purported to investigate the impact of interleukin-10 (IL-10) gene 4 polymorphisms (-1082G>A, -819T>C, -592A>C and 210T>C) on peripheral blood IL-10 variation and prostate cancer (PCa) risk, with a special consideration given to various origins of between-study heterogeneity. 2 researchers independently fulfilled literature retrieval, quality assessment and information collection. Sub-grouped analyses per ethnicity, continent, design type, control source, genotyping procedure, genotype validation, age-matched status, study sample size, quality score and controls’ mean age were conducted, respectively. Total 17 unduplicated studies (patients/controls: 7561/8101) were assessable for PCa risk, and 4 unduplicated studies (1189 subjects) for peripheral blood IL-10 variation. Pooling all assessable studies identified a marginally significant association between the -1082A allele and increased PCa risk (odds ratio (OR)=1.10, 95% confidence interval [CI]: 1.00 to 1.21) (Heterogeneity I 2 =64.3%), and no significance was detected in sub-grouped analyses of this polymorphism. Contrastingly, the -592C allele was significantly associated with reduced PCa risk in both prospective (OR=0.85, 95% CI: 0.77 to 0.95) and population-based (OR=0.92, 95% CI: 0.84 to 1.00) studies (Heterogeneity I 2 =0.0% and 18.1%). Moreover, carriers of combined -592CA/CC genotypes had a significant higher level of peripheral blood IL-10 than the -592AA genotype carriers (weighted mean difference=0.45 and 0.54 mg/dL, 95% CI: 0.23 to 0.67 and 0.30 to 0.39). The above comparisons possessed a low probability of publication bias. In sum, our findings suggested that IL-10 gene -592A>C polymorphism may represent a promising candidate locus for the occurrence of PCa, and further signified a contributing role of this polymorphism in prostate carcinogenesis.

// Geng Tian 1, * , Fangrong Tang 2, * , Chunhua Yang 1 , Wenfeng Zhang 2 , Jonas Bergquist 3 , Bin Wang 1 , Jia Mi 1, 3 and Jiandi Zhang 1, 2 1 Medicine and Pharmacy Research Center, Binzhou Medical University, Yantai, P. R. China 2 Yantai Zestern Biotechnique Co. LTD, Yantai, P. R. China 3 Department of Chemistry, BMC, Uppsala University, Uppsala, Sweden * These authors have contributed equally to this work Correspondence to: Jiandi Zhang, email: Jiandi.zhang@zestern.net Jia Mi, email: jiajiami@gmail.com Keywords: quantitative dot blot, high throughput, proteomics, immunoblot analysis, western blot Received: February 17, 2017 Accepted: March 26, 2017 Published: April 19, 2017 ABSTRACT Lacking access to an affordable method of high throughput immunoblot analysis for daily use remains a big challenge for scientists worldwide. We proposed here Q uantitative D ot B lot analysis (QDB) to meet this demand. With the defined linear range, QDB analysis fundamentally transforms traditional immunoblot method into a true quantitative assay. Its convenience in analyzing large number of samples also enables bench scientists to examine protein expression levels from multiple parameters. In addition, the small amount of sample lysates needed for analysis means significant saving in research sources and efforts. This method was evaluated at both cellular and tissue levels with unexpected observations otherwise would be hard to achieve using conventional immunoblot methods like Western blot analysis. Using QDB technique, we were able to observed an age-dependent significant alteration of CAPG protein expression level in TRAMP mice. We believe that the adoption of QDB analysis would have immediate impact on biological and biomedical research to provide much needed high-throughput information at protein level in this “Big Data” era.

// Fang Liu 1, 2, * , Yuan Zhang 1, * , Tingting Men 1 , Xingyue Jiang 2 , Chunhua Yang 1 , He Li 4 , Xiaodan Wei 1 , Dong Yan 1 , Gangming Feng 5 , Jianke Yang 1 , Jonas Bergquist 3 , Bin Wang 2 , Wenguo Jiang 1 , Jia Mi 1, 3 , Geng Tian 1 1 Medicine and Pharmacy Research Center, Binzhou Medical University, Yantai, Shandong Province, 264003 China 2 Department of Radiology, Affiliated Hospital of Binzhou Medical University, Binzhou, Shandong Province, 256603 China 3 Department of Chemistry – BMC, Uppsala University, Uppsala, 75124, Sweden 4 Department of Gastric and Intestine, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, Shandong Province, 264003 China 5 Yantai Institute, China Agriculture University, Yantai, Shandong Province, 264670 China * Shared first author Correspondence to: Geng Tian, email: Tiangeng@live.se Wenguo Jiang, email: jiangwg@foxmail.com Jia Mi, email: jia.mi@kemi.uu.se Keywords: proteomic, PDGF-B, gastric cancer, PRDX5, pathway Received: December 02, 2016 Accepted: February 07, 2017 Published: March 06, 2017 ABSTRACT Gastric cancer is one of the most common cancers in Asian countries. Searching for reliable biomarkers involving the development of gastric cancer is important for clinical practice. Quantitative proteomics has become an important method contributed to the discovery of novel diagnostic or therapeutic targets for the management of cancer. Here, we identified differently expressed proteins in gastric cancer and normal gastric tissues by using the high resolution mass spectrometer. Among the total of 2280 identified proteins, 87 were differentially expressed between gastric cancer and normal gastric tissues. Notably, several significant proteins are in the PDGF-B signaling pathway, including peroxiredoxin5 (PRDX5), S100A6, calreticulin (CALR) and cathepsin D (CTSD), which were validated by western blot. Furthermore, upstream regulators including PDGF-B, PDGFR-β, Akt, eIF4E and p70s6K were found significantly increased in the gastric cancer tissues. In addition, silencing of PRDX5 and PDGF-B suppressed the proliferation of gastric cancer cells in vitro . The administration of exogenous PDGF-BB recovered the reduced expression of PDGF-B signaling pathway in PDGF-B knockdown cells. Taken together, our findings suggested that PDGF-B signaling pathway plays an important role in the regulation of gastric cancer proliferation and the inhibition of this pathway may be a potential approach for treatment of gastric cancer.

Toxoplasma gondii is a ubiquitous, obligate intracellular parasite capable of crossing the placental barrier and causing spontaneous abortion, preterm labor, or significant disease in the surviving neonate. To better understand molecular mechanisms underlying abnormal pregnancy outcomes caused by T. gondii, placental proteins extracted from T. gondii-infected and -uninfected mice were comparatively analyzed using label-free liquid chromatography-tandem mass spectrometry. Significant difference was observed in the expression of 58 out of 792 proteins in infected placentas (p0.05) compared with that in uninfected placentas. Quantitative real-time polymerase chain reaction, western blotting, and immunohistochemical staining were used to validate the results of the proteomic analysis. Some placental proteins differentially expressed in infected and uninfected mice were found to be associated with several different biological processes of pregnancy, particularly with trophoblast invasion and placental development. The results provide possible novel insights into the molecular mechanisms for abnormal pregnancy outcomes associated with T. gondii infection.In order to further explore the mechanisms of abnormal pregnant outcomes caused by T. gondii infection, we first applied label-free proteomic technology to analyze the differentially expressed host placental proteins with T. gondii infection. The results showed that some differential proteins are associated with trophoblast invasion and placenta development. The findings provide a systemic view of the altered placental proteins and help to declare the molecular mechanisms of abnormal pregnancy outcomes caused by T. gondii infection.

A deeper understanding of how viruses reprogram their hosts for production of progeny is needed to combat infections. Most knowledge on the regulation of cellular gene expression during adenovirus infection is derived from mRNA studies. Here, we investigated the changes in protein expression during the late phase of adenovirus type 2 (Ad2) infection of the IMR-90 cell line by stable isotope labeling in cell culture with subsequent liquid chromatography-high resolution tandem mass spectrometric analysis. Two biological replicates of samples collected at 24 and 36 h post-infection (hpi) were investigated using swapped labeling. In total, 2648 and 2394 proteins were quantified at 24 and 36 hpi, respectively. Among them, 659 and 645 were deregulated1.6-fold at the two time points. The protein expression was compared with RNA expression using cDNA sequencing data. The correlation was surprisingly low (r = 0.3), and several examples of posttranscriptional regulation were observed; e.g., proteins related to carbohydrate metabolism were up-regulated at the protein level but unchanged at the RNA level, whereas histone proteins were down-regulated at the protein level but up-regulated at the RNA level. The deregulation of cellular gene expression by adenovirus is mediated at multiple levels and more complex than hitherto believed.

// Min Li 1,* , Peiyuan Wang 2,* , Chunhua Yang 1 , Wenguo Jiang 1 , Xiaodan Wei 1 , Xinbo Mu 3 , Xuri Li 1 , Jia Mi 1 and Geng Tian 1 1 Medicine and Pharmacy Research Center, Binzhou Medical University, Yantai, Shandong, China 2 Institute of Imaging, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, Shandong, China 3 Personnel Department, Binzhou Medical University, Yantai, Shandong, China * These authors have contributed equally to this work Correspondence to: Geng Tian, email: // Jia Mi, email: // Xuri Li, email: // Keywords : hepatitis C virus; chronic kidney disease; meta-analysis; effect estimate Received : August 19, 2016 Accepted : October 14, 2016 Published : October 25, 2016 Abstract We aimed to meta-analytically assess the predisposition of hepatitis C virus (HCV) infection to the occurrence and severity of chronic kidney disease (CKD). Two authors independently searched articles and abstracted information. Odds ratio (OR) or hazard ratio (HR) along with 95% confidence interval (CI) was converged separately in 12 longitudinal (1,972,044 subjects) and 15 cross-sectional (937,607 subjects) studies. Overall effect estimate was remarkably significant in longitudinal studies (HR, 95% CI, P : 1.45, 1.23-1.71, 95%). HCV infection was also associated with an 1.54-fold (95% CI, P : 1.27-1.87, < 0.001) increased risk of having prevalent proteinuria. In longitudinal studies with estimated glomerular filtration rate (eGFR) < 60, < 30 and < 15 ml/min/1.73m 2 , the corresponding HR was 1.39 (95% CI, P : 1.14-1.69, 0.001), 1.79 (0.91-3.51, 0.091) and 2.30 (1.26-4.19, 0.007). Further grouping the longitudinal studies by median follow-up time at 5 years revealed that the effect estimate was reinforced in long-term studies (HR, 95% CI, P : 1.86, 1.19-2.89, 0.006; I 2 =98.1%) relative to that in short-term studies (1.21, 1.03-1.43, 0.024; 92.0%). In conclusion, our findings demonstrate the significant risk of experiencing incident CKD after HCV infection, with the lower eGFR and longer HCV exposure time entailing a greater risk.

// Lei Zheng 1, 2, * , Chen Zhao 3, * , Yuxiang Du 2 , Xianchai Lin 2 , Yida Jiang 2 , Chunsik Lee 2 , Geng Tian 1 , Jia Mi 1 , Xianglin Li 1 , Qishan Chen 2 , Zhimin Ye 2 , Lijuan Huang 2 , Shasha Wang 2 , Xiangrong Ren 2 , Liying Xing 2 , Wei Chen 2 , Delong Huang 1, 2 , Zhiqin Gao 4 , Shuping Zhang 1 , Weisi Lu 2 , Zhongshu Tang 2 , Bin Wang 5 , Rong Ju 2 , Xuri Li 1, 2 1 Center for Medical and Pharmaceutical Research, Binzhou Medical University, Yantai, Shandong, 264003, P. R. China 2 State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, P. R. China 3 Department of Ophthalmology, the First Affiliated Hospital of Nanjing Medical University and State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing 210029, P. R. China 4 Department of Cell Biology, Weifang Medical University, Weifang, 261053 P. R. China 5 Medical Imaging Institute, Shandong Province Characteristical Key Subject, Medical Imaging and Nuclear Medicine, Binzhou Medical University, Yantai, 264003 P. R. China * Equal first authors Correspondence to: Xuri Li, email: lixr6@mail.sysu.edu.cn Rong Ju, email: jurong@mail.sysu.edu.cn Bin Wang, email: binwang001@aliyun.com Keywords: angiogenesis, PDGF-CC, VEGF-A, drug resistance Received: May 11, 2016 Accepted: October 14, 2016 Published: October 24, 2016 ABSTRACT Anti-VEGF-A therapy has proven to be effective for many neovascular diseases. However, drug resistance to anti-VEGF-A treatment can develop. Also, not all patients with neovascular diseases are responsive to anti-VEGF-A treatment. The mechanisms underlying these important issues remain unclear. In this study, using different model systems, we found that inhibition of VEGF-A directly upregulated PDGF-CC and its receptors in multiple cell types in pathological angiogenesis in vitro and in vivo . Importantly, we further revealed that combinatorial targeting of VEGF-A and PDGF-CC suppressed pathological angiogenesis more efficiently than monotherapy. Given the potent angiogenic activity of PDGF-CC, our findings suggest that the development of resistance to anti-VEGF-A treatment may be caused by the compensatory upregulation of PDGF-CC, and combined inhibition of VEGF-A and PDGF-CC may have therapeutic advantages in treating neovascular diseases.

To investigate how undergraduates use library and scholarly resources through their mobile devices, the authors surveyed 138 undergraduate students at The College of New Jersey (TCNJ) in the spring 2014 semester. The results confirmed that they want to search databases and journals on their mobile devices. By examining the functionalities of mobile-friendly databases, we identified the challenges and barriers to using mobile devices to search electronic databases and find articles. The results of this study were used in generating recommendations for the design, capabilities, and resources of future mobile websites.

With the recent energy crisis, the new energy harvesting technologies have become one of the hot spots in engineering academic research and industrial applications. By its wide range of application fields, vibration energy harvesting technologies have been gradually developed and utilized in which an efficient and stable harvester technology is one of the recent key problems. In order to improve energy harvesting efficiency and reduce energy loss caused by motor inertial commutation, many mechanical structures or hydraulic structures that convert reciprocating vibration energy into single direction rotation of motor are proposed. Although these methods can improve energy harvesting efficiency, they can have negative effects in some cases, especially in the case of vibration energy harvesting from human beings. This paper proposes a vibration harvesting mechanism with mechanical rectification filter function applied to backpack. The prototype model of the system was established in SolidWorks and imported into ADAMS. Thereafter, dynamic analyses of mechanical rectification filtering characteristics and meshing characteristics of one-way clutch were simulated in ADAMS. Based on ADAMS, parametric design analysis and its influence on the mechanical rectification characteristics were investigated. The simulation results were validated by bench test results. Simulation results is done by ADAMS and the results match well with bench test results.

The relationship between diet, especially polyphenols, and health has been receiving increasing attention. Polyphenols were degraded by gut microbiota into metabolites and acted on the body to exert many bioactivities from several targets such as antioxidative stress, anti-inflammation, intestinal barrier and gut microbiota modulation. After long-term treatment of mice with anthocyanins from Lycium ruthenicum Murray (ACN), antioxidant status in liver (T-AOC, T-SOD, CAT, GSH and GSH-Px were increased and AST, ALT, ALP and MDA were decreased), anti-inflammatory status in colon (the expression of mRNA of iNos, Cox-2, Tnf-α, Il-6, Il-1β and Ifn-γ were significantly reduced), intestinal barrier (the expression of mRNA of Zo-1, Occludin, Claudin-1 and Muc1 were significantly increased) and gut microbiota (Barnesiella, Alistipes, Eisenbergiella, Coprobacter and Odoribacter were proliferated) were all regulated in ACN group. Meanwhile, the content of short-chain fatty acids in cecal contents and feces were increased. Taken together, long-term intake of ACN could promote organism healthy and these results have important implications for the development of ACN as a functional food ingredient.

Zeaxanthin dipalmitate (ZDP) is a major non-saponified carotenoid in fully ripe fruits of Lycium barbarum L. In the present study, response surface methodology was used to optimize the ultrasonic-assisted extraction (UAE) conditions of carotenoids from the fruits of L. barbarum, and the optimal extraction conditions were determined as follows: ultrasonic power of 360 W, ultrasonic time of 40 min and the ratio of extraction solvent to sample of 30 mL/g. An actual value of ZDP content of 5.40 mg/g and short extraction time indicated the efficiency of UAE. Furthermore, a promising high-speed counter-current chromatography (HSCCC) method was established for the purification of ZDP from the fruits of L. barbarum. With a developed two-phase solvent system composed of n-hexane/dichloromethane/acetonitrile (10/3/7, v/v/v), ZDP with a purity of higher than 95% was successfully isolated from the crude extract. This is the first report on the purification of ZDP by using HSCCC.

Many studies have shown that pollen and its preparation are ideal herbal remedies for the treatment of prostate diseases. Our previous study found that pollen polysaccharides from Chinese wolfberry (WPPs) can induce the apoptosis of prostate cancer DU145 cells. But the antitumor mechanism of WPPs was not clearly understood. Therefore, in the present study, we further investigated the antitumor mechanism of WPPs in DU145 cells and a xenograft mice model. The results showed that WPPs decreased the levels of PI3K, AKT, p-AKT and Bcl-2 proteins, and increased expression of Bax, caspase-3 and caspase-9 in DU145 cells (P

In the present study, the effects of Lycium barbarum polysaccharides (LBPS) on immunoregulation and gut microbiota dysbiosis in cyclophosphamide (CTX)-induced mice were investigated to elucidate whether the attenuation of immunosuppression is related to the modulation of the gut microbiota. The results showed that administration of LBPS could protect immune organs (enhancing immune organ indexes and alleviating immune organ damage), enhance the production of immune-related cytokines (IL-2, IL-6, IL-1β, TNF-α and IFN-γ) and prevent the hepatotoxicity in CTX-induced mice. Additionally, LBPS treatment could promote the production of short-chain fatty acids and modulate the composition of the gut microbiota, increasing the relative abundances of Bacteroidaceae, Lactobacillaceae, Prevotellaceae and Verrucomicrobiaceae, which were positively associated with immune traits. The present results indicated that LBPS might regulate the immune response depending on the modulation of the gut microbiota, suggesting that LBPS could be developed as special ingredients for immunoregulation in association with the modulation of the gut microbiota.

Purpose Activation of the hypothalamus-pituitary-adrenal (HPA) axis pathway is closely related to insulin resistance (IR), glucose, and lipid metabolism disorders in type 2 diabetes mellitus (T2DM). Berberine (BBR) has effect on regulating disorder of glucose and lipid metabolism in T2DM. In fact, activation of the HPA axis pathway is closely related to IR, glucose, and lipid metabolism disorders in T2DM. Here, we investigated whether the therapeutic effect of BBR on T2DM rats is acted through the HPA axis pathway. Methods In this research, we investigated the effects of BBR on the HPA-axis pathway-related indicators and expression of skeletal muscle glucose transporter 4 (GLUT4) in the high-fat diet and streptozotocin-induced T2DM rats, and identify its possible mechanism of improving IR in T2DM. Results BBR significantly reduced fasting blood glucose, total cholesterol, and low-density lipoprotein cholesterol in model rats. It also improved the abnormalities of the high-density lipoprotein cholesterol, the insulin resistance index, the insulin sensitivity index, glucagon, and insulin levels. BBR decreased levels of hypothalamic Orexin-A, the OX2R receptor, the corticotropin-releasing hormone, the pituitary and the plasma adrenocorticotropic hormone, as well as serum and urine corticosterone. At the same time, BBR increased mRNA and protein expressions of GLUT4 in skeletal muscles of model rats as well. Conclusion Those results suggested that BBR can exert inhibition on the HPA-axis and increased skeletal muscle expression of GLUT4 proteins, which may be one of the important mechanisms in BBR to improve IR and regulating glucose and lipid metabolism in T2DM rats.

To circumvent the limitations associated with sandwich ELISA for tissue biomarker quantitation, Quantitative Dot Blot method (QDB) was proposed using antibodies clinically validated for immunohistochemistry (IHC), as this method requires only one primary antibody in the analysis. The protein levels of four breast cancer tissue biomarkers, including Estrogen Receptor (ER), Progesterone Receptor (PR), Ki67 and Her2, were absolutely quantitated successfully in 190 frozen breast tissue biopsies, and the results were further verified with provided IHC results. We propose QDB method as an alternative platform to Sandwich ELISA for absolute quantitation of tissue biomarkers with significantly reduced developing effort and time.

In the present study, the therapeutic effects of crude anthocyanins (ACN) from the fruits of Lycium ruthenicum Murray and the main monomer of ACN, petunidin 3-O-[rhamnopyranosyl-(trans-p-coumaroyl)]-5-O-[β-d-glucopyranoside] (P3G), on the dextran sodium sulfate (DSS)-induced colitis in mice were investigated. Both ACN and P3G showed intestinal anti-inflammatory effects, evidenced by restoration of various physical signs (body weight, feed quantity, solid fecal weight and colon length were increased, and DAI score was decreased), reduction of the expression of proinflammatory cytokines and related mRNA (such as TNF-α, IL-6, IL-1β and IFN-γ), and promotion of the intestinal barrier function by histological and immunofluorescence analysis (proteins such as ZO-1, occludin and claudin-1 were increased). Furthermore, the effects on gut microbiota community of DSS-induced colitis in mice have been investigated. It was found that Porphyromonadaceae, Helicobacter, Parasutterella, Parabacteroides, Oscillibacter and Lachnospiraceae were the key bacteria associated with inflammatory bowel disease. Taken together, P3G and ACN ameliorated DSS-induced colitis in mice through three aspects including blocking proinflammatory cytokines, increasing tight junction protein and modulating gut microbiota. What's more, P3G showed better anti-inflammatory effects than ACN.

Both Lycium barbarum L. polysaccharides (LBP) and zinc have protective effects on liver injuries. In this paper, LBP and ZnSO4 were combined to study the effects on the prevention of alcoholic liver injury. The rats were divided into six groups, the normal group, alcohol group, zinc sulfate group, LBP group, low‐dose group of ZnSO4, and high‐dose group of ZnSO4 and LBP, used to explore the impact of LBP and ZnSO4 complex on liver lipid metabolism of alcohol, alcohol‐metabolizing enzymes, oxidative damage, and inflammation of the liver. The experimental model was established by gavage treatment, observation, and determination of indexes of rats. The results showed that the combination of LBP and ZnSO4 could significantly decrease the levels of triglyceride (TG), total cholesterol (TC), tumor necrosis factor‐α(TNF‐ɑ), malondialdehyde (MDA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and the activity of enzyme subtype 2E1 (CYP2E1). It also significantly increased the activities of total superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH‐Px), glutathione peptide (GSH), and alcohol dehydrogenase, effectively improved the liver tissue lesion. What is more, the combination of LBP and ZnSO4 had a synergistic effect on the remission of alcoholic fatty liver, and alleviated chronic alcoholic liver injury by promoting lipid metabolism, inhibiting oxidative stress, controlling inflammatory responses, and regulating the expression and activity of alcohol‐metabolizing enzymes in rats.

Background Esophageal cancer (EC) is one of the malignant tumors with a poor prognosis. The early stage of EC is asymptomatic, so identification of cancer biomarkers is important for early detection and clinical practice. Methods In this study, we compared the protein expression profiles in esophageal squamous cell carcinoma (ESCC) tissues and adjacent normal esophageal tissues from five patients through high-resolution label-free mass spectrometry. Through bioinformatics analysis, we found the differentially expressed proteins of ESCC. To perform the rapid identification of biomarkers, we adopted a high-throughput protein identification technique of Quantitative Dot Blot (QDB). Meanwhile, the QDB results were verified by classical immunohistochemistry. Results In total 2297 proteins were identified, out of which 308 proteins were differentially expressed between ESCC tissues and normal tissues. By bioinformatics analysis, the four up-regulated proteins (PTMA, PAK2, PPP1CA, HMGB2) and the five down-regulated proteins (Caveolin, Integrin beta-1, Collagen alpha-2(VI), Leiomodin-1 and Vinculin) were selected and validated in ESCC by Western Blot. Furthermore, we performed the QDB and IHC analysis in 64 patients and 117 patients, respectively. The PTMA expression was up-regulated gradually along the progression of ESCC, and the PTMA expression ratio between tumor and adjacent normal tissue was significantly increased along with the progression. Therefore, we suggest that PTMA might be a potential candidate biomarker for ESCC. Conclusion In this study, label-free quantitative proteomics combined with QDB revealed that PTMA expression was up-regulated in ESCC tissues, and PTMA might be a potential candidate for ESCC. Since Western Blot cannot achieve rapid and high-throughput screening of mass spectrometry results, the emergence of QDB meets this demand and provides an effective method for the identification of biomarkers.