Exercise accelerates recruitment of CD8+ T cell to promotes anti-tumor immunity in lung cancer via epinephrine.

BACKGROUND AND PURPOSE: In recent years, there has been extensive research on the role of exercise as an adjunctive therapy for cancer. However, the potential mechanisms underlying the anti-tumor therapy of exercise in lung cancer remain to be fully elucidated. As such, our study aims to confirm whether exercise-induced elevation of epinephrine can accelerate CD8+ T cell recruitment through modulation of chemokines and thus ultimately inhibit tumor progression. METHOD: C57BL/6 mice were subcutaneously inoculated with Lewis lung cancer cells (LLCs) to establish a subcutaneous tumor model. The tumor mice were randomly divided into different groups to performed a moderate-intensity exercise program on a treadmill for 5 consecutive days a week, 45 min a day. The blood samples and tumor tissues were collected after exercise for IHC, RT-qPCR, ELISA and Western blot. In addition, another group of mice received daily epinephrine treatment for two weeks (0.05 mg/mL, 200 µL i.p.) (EPI, n = 8) to replicate the effects of exercise on tumors in vivo. Lewis lung cancer cells were treated with different concentrations of epinephrine (0, 5, 10, 20 µM) to detect the effect of epinephrine on chemokine levels via ELISA and RT-qPCR. RESULTS: This study reveals that both pre- and post-cancer exercise effectively impede the tumor progression. Exercise led to an increase in EPI levels and the infiltration of CD8+ T cell into the lung tumor. Exercise-induced elevation of EPI is involved in the regulation of Ccl5 and Cxcl10 levels further leading to enhanced CD8+ T cell infiltration and ultimately inhibiting tumor progression. CONCLUSION: Exercise training enhance the anti-tumor immunity of lung cancer individuals. These findings will provide valuable insights for the future application of exercise therapy in clinical practice.

Chanling Gao suppresses colorectal cancer via PI3K/Akt/mTOR pathway modulation and enhances quality of survival.

The Chinese medicine formula Chanling Gao (CLG) exhibits significant tumor inhibitory effects in colorectal cancer (CRC) nude mice. However, the detailed mechanisms remain elusive. CRC in situ nude mouse models were treated with CLG. Small animal magnetic resonance imaging (MRI) tracked tumor progression, and overall health metrics such as food and water intake, body weight, and survival were monitored. Posttreatment, tissues and blood were analyzed for indicators of tumor inhibition and systemic effects. Changes in vital organs were observed via stereoscope and hematoxylin-eosin staining. Immunohistochemistry quantified HIF-1α and P70S6K1 protein expression in xenografts. Double labeling was used to statistically analyze vascular endothelial growth factor (VEGF) and CD31 neovascularization. Enzyme-linked immunosorbent assay was used to determine the levels of VEGF, MMP-2, MMP-9, IL-6, and IL-10 in serum, tumors, and liver. Western blotting was used to assess the expression of the PI3K/Akt/mTOR signaling pathway-related factors TGF-ß1 and smad4 in liver tissues. CLG inhibited tumor growth, improved overall health metrics, and ameliorated abnormal blood cell counts in CRC nude mice. CLG significantly reduced tumor neovascularization and VEGF expression in tumors and blood. It also suppressed HIF-1α, EGFR, p-PI3K, Akt, p-Akt, and p-mTOR expression in tumors while enhancing PTEN oncogene expression. Systemic improvements were noted, with CLG limiting liver metastasis, reducing pro-inflammatory cytokines IL-6 and IL-10 in liver tissues, decreasing MMP-2 in blood and MMP-2 and MMP-9 in tumors, and inhibiting TGF-ß1 expression in liver tissues. CLG can enhance survival quality and inhibit tumor growth in CRC nude mice, likely through the regulation of the PI3K/Akt/mTOR signaling pathway.

Deubiquitinating PABPC1 by USP10 upregulates CLK2 translation to promote tumor progression in pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is extremely malignant with limited treatment options. Deubiquitinases (DUBs), which cleave ubiquitin on substrates, can regulate tumor progression and are appealing therapeutic targets, but there are few related studies in PDAC. In our study, we screened the expression levels and prognostic value of USP family members based on published databases and selected USP10 as the potential interventional target in PDAC. IHC staining of the PDAC microarray revealed that USP10 expression was an adverse clinical feature of PDAC. USP10 promoted tumor growth both in vivo and in vitro in PDAC. Co-IP experiments revealed that USP10 directly interacts with PABPC1. Deubiquitination assays revealed that USP10 decreased the K27/29-linked ubiquitination level of the RRM2 domain of PABPC1. Deubiquitinated PABPC1 was able to couple more CLK2 mRNA and eIF4G1, which increased the translation efficiency. Replacing PABPC1 with a mutant that could not be ubiquitinated impaired USP10 knock-down-mediated tumor suppression in PDAC. Targeting USP10 significantly delayed the growth of cell-derived xenograft and patient-derived xenograft tumors. Collectively, our study first identified USP10 as the DUB of PABPC1 and provided a rationale for potential therapeutic options for PDAC with high USP10 expression.

Inhibition of epithelial-to-mesenchymal transition augments antitumor efficacy of nanotherapeutics in pancreatic ductal adenocarcinoma.

Intrinsic drug resistance mechanisms of tumor cells often reduce intracellular drug concentration to suboptimal levels. Epithelial-to-mesenchymal transition (EMT) is a pivotal process in tumor progression and metastasis that confers an aggressive phenotype as well as resistance to chemotherapeutics. Therefore, it is imperative to develop novel strategies and identify new targets to improve the overall efficacy of cancer treatment. We developed SN38 (active metabolite of irinotecan)-assembled glycol chitosan nanoparticles (cSN38) for the treatment of pancreatic ductal adenocarcinoma (PDAC). Furthermore, cSN38 and the TGF-ß1 inhibitor LY364947 formed composite nanoparticles upon self-assembly (cSN38 + LY), which obviated the poor aqueous solubility of LY364947 and enhanced drug sensitivity. The therapeutic efficacy of cSN38 + LY nanotherapeutics was studied in vitro and in vivo using suitable models. The cSN38 nanoparticles exhibited an antitumor effect that was significantly attenuated by TGF-ß-induced EMT. The cellular uptake of SN38 was impeded during EMT, which affected the therapeutic efficacy. The combination of LY364947 and cSN38 markedly enhanced the cellular uptake of SN38, increased cytotoxic effects, and inhibited EMT in PDAC cells in vitro. Furthermore, cSN38 + LY significantly inhibited PDAC xenograft growth in vivo. The cSN38 + LY nanoparticles increased the therapeutic efficacy of cSN38 via repressing the EMT of PDAC cells. Our findings provide a rationale for designing nanoscale therapeutics to combat PDAC.

SIGLEC15 amplifies immunosuppressive properties of tumor-associated macrophages in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) usually presents infrequent infiltration of T lymphocytes. The known immune-checkpoint inhibitors to date focus on activating T cells and manifest limited effectiveness in PDAC. SIGLEC15 was identified as a novel tumor-associated macrophage (TAM)-related immune-checkpoint in other cancer types, while its immunosuppressive role and clinical significance remained unclear in PDAC. In our study, SIGLEC15 presented immunosuppressive relevance in PDAC via bioinformatic analysis and expressed on TAM and PDAC cells. SIGLEC15+ TAM, rather than SIGLEC15+ PDAC cells or SIGLEC15- TAM, correlated with poor prognosis and immunosuppressive microenvironment in the PDAC microarray cohort. Compared with SIGLEC15- TAM, SIGLEC15+ TAM presented an M2-like phenotype that could be modulated by SIGLEC15 in a tumor cell-dependent manner. In mechanism, SIGLEC15 interacted with PDAC-expressed sialic acid, preferentially α-2, 3 sialic acids, to stimulate SYK phosphorylation in TAM, which further promoted its immunoregulatory cytokines and chemokines production. In vivo, SIGLEC15+ TAM also presented an M2-like phenotype, accelerated tumor growth, and facilitated immunosuppressive microenvironment, which was greatly abolished by SYK inhibitor. Our study highlighted a novel M2-promoting function of SIGLEC15 and strongly suggested SIGLEC15 as a potential immunotherapeutic target for PDAC.

First Report of Bipolaris cactivora Causing Flower Rot of Pitaya (Hylocereus costaricensis) in China.

Pitaya (Hylocereus costaricensis), belonging to the Cactaceae family, has rich functional substances, such as a variety of amino acids, which are popular with consumers (Wichienchot et al. 2010). In May 2019, flowers showed symptoms of rot, with an incidence of 15% in a plantation (233.3 ha) in Changjiang (19°46'N; 108°93'E) (Hainan province), China. The initial disease symptoms of flower were small scattered purple-red spot (1~2 mm), including circular, long oval or irregular in shape. The spots were gradually expanded and coalesced, forming abundant reddish-brown lesions. Later, this disease resulted in rotting and blackening of the whole flower. Many black mildew layers (conidiophores and conidia) on the surface of the lesions were observed under compound microscopy. Symptomatic flower tissue (4 cm2) from collecting samples was disinfected in 75% ethanol for 25 s, followed by 1 min in 5% sodium hypochlorite, rinsed 3 times with sterile water, plated on potato dextrose agar (PDA) for 3 days, and incubated at 28ºC. A fungus was consistently isolated from symptomatic flower samples with 90% isolation rate. Resultant colony of the fungus was circular, dark green, velvety, hairy, after 7 days, incubated at 28ºC. Hyphae were septate, 6.2-8.9 µm (average 7.6±0.5) in diameter. Conidia were straight, obclavate, pale to mid brown, 2-6 septate, 23.0 to 42.2 µm (average 31.0±3.2) × 6.5 to 9.8 µm (average 8.0±0.6) (n = 100). The conidia were normally produced germ tubes from one end or both ends. The width of conidiophore was 5.1 to 6.6 µm (average 5.8±0.4) (n = 50). Sequences were generated from the isolate using primers for the internal transcribed spacer region (ITS) (ITS1/ITS4) (White et al. 1990), ribosomal large subunit (LSU) (LROR/LR5) (Vilgalys et al. 1990), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (GPD1/GPD2) (Berbee et al. 1999) loci. The resulting sequences were deposited in GenBank with accession numbers MN960109, MN966852, and MT542865. BLAST analysis demonstrated that these sequences were 99% similar to ITS (HM193535), LSU (MH869295), and GAPDH (HM598681) of Bipolaris cactivora. A maximum likelihood phylogenetic analysis based on combined dataset of ITS, LSU, and GAPDH sequences using MEGA7.0 revealed that the isolate was placed in the same clade as B. cactivora with 100% bootstrap support. A conidial suspension (1 × 105 conidia/ml) of the fungal isolate was prepared by harvesting conidia from pure culture of the fungus grown on PDA 25 days. The 10 mL suspension was sprayed onto ten flowers with no wounding. Ten additional flowers sprayed with sterile distilled water were served as controls. All flowers were covered with plastic bags to maintain high humidity and incubated under natural condition. Typical symptoms of purple-red spot were observed on all the inoculated flowers on the third day. Abundant dark-brown to dark lesions were observed on the surface of flowers and were similar to those observed on the naturally infected flowers after 5 days. The control flowers remained asymptomatic. The fungal isolate of B. cactivora was reisolated from lesion of the flowers and reidentified by morphological and molecular characteristics, thus fulfilled Koch's postulates. Pathogenicity tests were repeated thrice with the same results. B. cactivora had been reported causing flowers and fruit rot of pitaya in South Florida (Tarnowski et al. 2010). This is the first report of B. cactivora causing flower rot of pitaya (H. costaricensis) in China. The flower rot may provide inoculum for the fruit rot, which will cause reduction of pitaya yield.

Oncological outcomes of laparoscopic versus open surgery in pT4 colon cancers: A systematic review and meta-analysis.

BACKGROUND: Widespread adoption of minimally invasive surgery for colon cancer has achieved improved short-term benefits and better long-term oncological outcomes compared with open surgery. However, it is still controversial whether laparoscopic surgery is suitable for patients with stage T4 colon cancer. The aim of this meta-analysis was to compare short- and long-term oncological outcomes associated with laparoscopic and conventional open surgery for pT4 colon cancer. METHODS: Published studies from 2003 to 2018 comparing oncological outcomes following laparoscopic and open surgery for pT4 colon cancer were systematically searched. Data on conversion rate, R0 resection rate, number of harvested lymph nodes, morbidity and mortality, and overall survival (OS) and disease-free survival (DFS) were subjected to meta-analysis using fixed-effect and random-effect models. RESULTS: Twelve observational studies met the inclusion criteria with a total of 2396 cases (1250 laparoscopic and 1146 open). There was no significant difference in R0 resection rate [relative risk (RR) = 1.007; 95% confidence interval (CI) = 0.935-1.085; P = 0.850], number of harvested lymph nodes (MD = 0.004; 95% CI = -0.139 to 0.148; P = 0.951), mortality (RR = 0.509; 95% CI = 0.176-1.470; P = 0.212), and 3-year OS (RR = 1.056; 95% CI = 0.939-1.188; P = 0.360), 5-year OS (RR = 1.003; 95% CI = 0.883-1.139; P = 0.966), 3-year DFS (RR = 1.032; 95% CI = 0.903-1.179; P = 0.642), and 5-year DFS (RR = 0.995; 95% CI = 0.868-1.140; P = 0.973) between the groups. The rate of conversion from laparoscopic to open procedures was 10.7% (95% CI = 0.090-0.124). There was a significant difference in incidence of complications within 30 postoperative days between laparoscopic and open surgery (RR = 0.703; 95% CI = 0.564-0.876; P = 0.002). CONCLUSION: Laparoscopic surgery is safe and feasible in pT4 colon cancer, oncological outcomes are similar, and more importantly, there are fewer postoperative complications compared with open surgery.

Convolutional Neural Network Based on Extreme Learning Machine for Maritime Ships Recognition in Infrared Images.

The success of Deep Learning models, notably convolutional neural networks (CNNs), makes them the favorable solution for object recognition systems in both visible and infrared domains. However, the lack of training data in the case of maritime ships research leads to poor performance due to the problem of overfitting. In addition, the back-propagation algorithm used to train CNN is very slow and requires tuning many hyperparameters. To overcome these weaknesses, we introduce a new approach fully based on Extreme Learning Machine (ELM) to learn useful CNN features and perform a fast and accurate classification, which is suitable for infrared-based recognition systems. The proposed approach combines an ELM based learning algorithm to train CNN for discriminative features extraction and an ELM based ensemble for classification. The experimental results on VAIS dataset, which is the largest dataset of maritime ships, confirm that the proposed approach outperforms the state-of-the-art models in term of generalization performance and training speed. For instance, the proposed model is up to 950 times faster than the traditional back-propagation based training of convolutional neural networks, primarily for low-level features extraction.

An applied anatomical study of bronchial artery.

The aim of this study was to reveal the external features of the bronchial artery (BA) system, so as to provide morphological basis for clinic. The BAs in 48 adult cadavers were dissected and analyzed. The number of BAs in 48 cases was 118. The incidence of BA arising from thoracic aorta, right posterior intercostal artery, and right subclavian artery was 69.49, 27.12, and 3.39%, respectively. The origin of BAs in individual specimen might be single, two, or all of them, respectively. According to the different origin and/or origins of BAs, it could be divided into five categories. As for the course of BAs, in this study, all the left BAs arising from thoracic aorta passed forward around the left side of esophagus and then entered left pulmonary hilum; most (n = 15) of the right BAs arising from thoracic aorta passed forward around the left side of esophagus and then entered right pulmonary hilum; a few (n = 8) of the right BAs arising from thoracic passed forward the right side of esophagus and bronchus and then entered right pulmonary hilum. Besides, in our group, the special courses were that right intercostal-bronchial trunk (RICBT) arising from thoracic aorta passed between vertebra and esophagus and gave off BA which curved forward around the right side of esophagus and then entered right pulmonary hilum, common bronchial trunk (CBT) arising from thoracic aorta passed forward around the left side of esophagus laying anterior to bronchus or posterior to bronchus, then dividing into a left and a right BAs entering right and left pulmonary hilum, respectively. In 4 cadavers, the RICBT gave off the radiculomedullary artery and BA in turn, so radiculomedullary artery has the same origin with BA. Of all BAs, the mean diameter of right posterior intercostal artery, CBT, left BA, and right BA was 2.17 ± 0.84, 1.79 ± 0.57, 1.44 ± 0.50, and 1.39 ± 0.38 mm, respectively. The information gained from this study will be of value in clinic application.

Bone marrow mesenchymal stem cells improve learning ability of the aging rat / 中国组织工程研究

BACKGROUND: At present, studies have shown that bone marrow mesenchymal stem cells (BMSCs) have self-renewal ability, which can be used as ideal seed cells for repairing tissue and organ damages caused by aging and lesions. OBJECTIVE: To study the changes in the levels of oxidation, inflammatory factors and neurotrophic factors (BDNF) in the brain of aging rats undergoing BMSCs transplantation, and to analyze the mechanism underlying the repair of learning and memory ability in the aging rats. METHODS: A total of 30 clean Sprague-Dawley rats were randomly divided into control group, model group and BMSCs group, 10 rats in each group. Aging models were made in the rats by 3-month subcutaneous injection of D-galactose. After modeling, BMSCs treatment was performed via tail vein injection in the BMSCs group.The injection was performed once a week,for 8 continuous weeks.Morris water maze was used to detect the learning and memory abilities of the rats in each group after the final injection of BMSCs. Superoxide dismutase activity in the brain tissue of rats was detected by xanthine oxidase method. Level of malondialdehyde in the rat brain tissue was detected by thiobarbituric acid method. Total antioxidant capacity of the brain tissue was detected by Fe3+reduction method. Real-time PCR and western blot assay were used to detect the expression of brain-derived neurotrophic factor mRNA and protein in the brain tissue of the aging rat, respectively. RESULTS AND CONCLUSION: Compared with the model group, the BMSCs group exhibited significantly higher activity of superoxide dismutase, stronger total antioxidant capacity, and higher levels of brain-derived neurotrophic factor mRNA and protein (P < 0.05), but the lower malondialdehyde level in the brain (P < 0.05). Compared with the model group, there was less time and higher frequency for passing through the platform in the BMSCs group (P < 0.05). Our findings further indicate that BMSCs can improve the abilities of learning and memory in aging rats, and the underlying mechanism is likely to improve antioxidant capacity and to regulate the level of brain-derived neurotrophic factors.

GC analysis of black gel pen ink stored under different conditions.

In many criminal and civil cases in China, the most commonly questioned documents are those written with gel pen ink. An important task for forensic document examiners is to identify whether two or more ink entries in one or more documents were written with the same ink type. The identification of the age of gel ink entries made poses an important and difficult problem for forensic document examiners. In this paper, the volatile components of gel ink were determined and the gel ink was classified by gas chromatography with a flame ionization detector. Calibration curves were created to express the relationship between the content of volatile gel ink components and the age of gel ink entries stored under natural and UV-induced aging conditions. The correspondence between the natural and UV-induced aging conditions was also established. The experimental results showed that GC was useful in the analysis of black gel ink and applicable for determining the relative age of gel ink entries under certain conditions.

[Research advances in anaerobic co-digestion of biogas fermentation substrates].

With global climate change, more and more attention has been paid to the development of bio-energy. Biogas fermentation, as a fairly mature technology of bio-matter energy transformation, has received considerable attention and experienced much development. How to improve the efficiency of biogas fermentation and promote its industrialization is a pressing issue. Anaerobic co-digestion is a simple, low-cost, and high-efficiency method for enhancing the efficiency of biogas fermentation, and received increasing attention from related researchers. This paper summarized the characteristics of various fermentation substrates, reviewed the research advances in the co-digestion of animal manure, sewage sludge, and industrial waste, with the focus on the advantages of co-digestion and the factors affecting the rate and efficiency of co-digestion, and prospected the future research of co-digestion and its application, aimed to provide theoretical guidance for the promotion and application of co-digestion techniques.

Investigation of thrombospondin-1 and transforming growth factor-ß expression in the heart of aging mice.

Elderly patients face the problems of morbidity and mortality due to age-mediated disabilities. The purpose of the present study was to investigate the expression of thrombospondin-1 (TSP-1) and transforming growth factor-ß (TGF-ß) in aging mice, and its probable mechanism in the pathological changes of aging myocardium. The aging model group (AM) comprised 30-month-old mice, while the control group comprised 2-month-old mice. The pathological changes were explored by H&E staining, and the contents of superoxide dismulase (SOD) and malondialdehyde (MDA) in the hearts were determined by xanthine oxidation or TBA colorimetry. TSP-1 and TGF-ß expression in the left ventricular myocardium was also measured by immunohistochemistry. The results showed that the activities of SOD decreased and the MDA content increased markedly in the hearts of the AM group compared to the control group. H&E staining showed that the control group myocardial cells lined up in order with clear structure and stained equably, while the AM group myocardial cells lined up in disorder with an augmented cell body and the appearance of many granules and interstitial fibrosis. Compared to the control group, in the hearts of the AM group, TSP-1 and TGF-ß protein expression in myocardial cells showed a significant increase (P<0.01). TSP-1 and TGF-ß expression increased in the myocardium, which may be related to pathological changes of age-related heart diseases, such as hypertrophy, fibrosis of myocardial cells and microvessel dissepiment thickening.

The association between two single nucleotide polymorphisms within the insulin-degrading enzyme gene and Alzheimer's disease in a Chinese Han population.

Several previous studies on the relationship between the insulin-degrading enzyme (IDE) gene and Alzheimer's disease (AD) have connected certain genetic variants to late-onset AD, in the absence of the apolipoprotein E (APOE)ε4 allele. However, the conclusions of these studies remain controversial. We investigated the association between two polymorphisms of IDE with AD in the Chinese population and found that the T/A genotype of rs4646958 had an important role in AD (adjusted p=0.007, odds ratio [OR]=2.796, 95% confidence interval [CI]=1.330-5.878), under the co-dominant genetic model. The T/C genotype of rs1887922 was also significantly associated with AD compared to the T/T genotype (adjusted p=0.003, OR=2.644, 95% CI=1.407-4.970). The C allele of rs1887922 conferred a higher risk of AD under the dominant genetics model (adjusted p=0.001, OR=2.719, 95% CI=1.472-5.022). Compared with the two other variant genotypes, the T/T genotype showed a protective effect in both polymorphisms (adjusted p=0.007, OR=0. 358, 95% CI=0.170-0.752 for rs4646958; adjusted p=0.001, OR=0. 368, 95% CI=0.199-0.679 in rs1887922). In the context of APOEε4-negative status, both variants were significantly associated with AD in some genetic models.

Association study of clusterin polymorphism rs11136000 with late onset Alzheimer's disease in Chinese Han population.

OBJECTIVE: We conducted a case-control study to investigate whether clusterin polymorphism (rs11136000) was associated with late-onset Alzheimer's disease in Chinese Han population. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was performed on genotype rs11136000 and APOEε4 in 127 patients with late-onset Alzheimer's disease and 143 control individuals. Previous published data from other Chinese samples was also included for further meta-analysis. RESULTS: APOEε4 was demonstrated to increase the risk of Alzheimer's disease in Chinese population (odds ratio = 2.35, 95% confidence interval: 1.40-3.96). There is no significant association between clusterin rs11136000 with late-onset sporadic AD in our small cohort. However, meta-analysis revealed significant allele and genotype differences between Alzheimer's disease and controls following a recessive model. CONCLUSION: Clusterin (rs11136000) was associated with Alzheimer's disease in Chinese Han population.

Lack of association between ATP13A2 Ala746Thr variant and Parkinson's disease in Han population of mainland China.

ATP13A2 (PARK9) mutations are related to Kufor-Rakeb syndrome (KRS). We performed genetic analysis of the Ala746Thr variant in an independent cohort of the patients with PD and healthy controls from mainland China. The Ala746Thr variant was present in 1/532 (0.19%) of PD compared with 1/480 (0.21%) of healthy controls (odds ratio=0.90, 95% CI 0.06, 14.39, P=1.00). The two subjects carried the heterozygous genotype. Subset analysis in the group 50 years of age showed 0% in PD versus 0.3% in healthy controls. We did not observe a significant association between Ala746Thr and Parkinson's disease in Han Chinese population, even after stratification by age at onset. The results suggested that Ala746Thr variant was not a major susceptible factor for PD in Han Chinese people.

[An evaluation of net carbon sink effect and cost/benefits of a rice-rape rotation ecosystem under long-term fertilization from Tai Lake region of China].

Taking a long-term fertilized rice-rape rotation system in Taihu Lake as test objective, its annual C balance and economic benefit were estimated, based on the measurement of past years grain yield, litter C content, and field CO2 emission as well as the investigation of material and management inputs. The calculated annual C sink under different fertilizations ranged from 0.9 t C x hm(-2) x a(-1) to 7.5 t C x hm(-2) x a(-1), and the net C sink effect under combined inorganic/organic fertilization was three folds as that under chemical fertilization. The C cost of material input ranged from 0.37 t C x hm(-2) x a(-1) to 1.13 t C x hm(-2) x a(-1), and that of management input ranged from 1.69 t C x hm(-2) x a(-1) to 1.83 t C x hm(-2) x a(-1). The annual economic benefit ranged from 5.8 x 10(3) CNY x hm(-2) x a(-1) to 16.5 x 10(3) CNY x hm(-2) x a(-1), and was 2.1 times higher under combined fertilization than under chemical fertilization. Comparing with that under chemical fertilization, the marginal cost for per ton C sink under combined inorganic/organic fertilization was estimated as 217.1 CNY x t(-1) C, very close to the C price of 20 Euro x t(-1) C in the EU. In sum, under combined inorganic/organic fertilization, this rice paddy ecosystem could not only have higher productivity, but also present greater net C sink effect and higher economic benefit, compared with under chemical fertilizer fertilization.

Designer self-assembling peptide scaffold stimulates pre-osteoblast attachment, spreading and proliferation.

A new peptide scaffold was made by mixing pure RADA16 (Ac-RADARADARADARADA-CONH2) and designer peptide RGDA16 (Ac-RADARGDARADARGDA-CONH2) solutions, and investigate any effect on attachment, spreading and proliferation of pre-osteoblast (MC3T3-E1). The peptides, RADA16 and RGDA16, were custom-synthesized. They were solubilized in deionized water at a concentration of 10 mg/ml (1% w/v), the RGDA16 peptide solution was mixed 1:1 with RADA16 solution and a new peptide solution RGDAmix was produced. The RGDAmix and RADA16 solution were directly loaded in 96-well plates and cover slips, and two different peptide scaffolds were formed with the addition of maintenance medium (alpha-MEM) in several minutes. About 1.0 x 10(4) MC3T3-E1 cells were seeded on each hydrogel scaffold, and then the cell morphological changes were observed using a fluorescence microscope at 1 h, 3 h and 24 h timepoint, respectively. Cell attachment was evaluated 1 h, 3 h and 24 h after cell seeding and cell proliferation was determined 4d, 7d and 14d after cell seeding. The RGDAmix scaffold significantly promoted the initial cell attachment compared with the RADA16 scaffold. MC3T3-E1 cells adhered and spread well on both scaffolds, however, cells spread better on the RGDAmix scaffold than on the RADA16 scaffold. Cell proliferation was greatly stimulated when cultured on RGDAmix scaffold. The RGD sequence contained peptide scaffold RGDAmix significantly enhances MC3T3-E1 cells attachment, spreading and proliferation.

Concentration-dependent modulation of amyloid-beta in vivo and in vitro using the gamma-secretase inhibitor, LY-450139.

LY-450139 is a gamma-secretase inhibitor shown to have efficacy in multiple cellular and animal models. Paradoxically, robust elevations of plasma amyloid-beta (Abeta) have been reported in dogs and humans after administration of subefficacious doses. The present study sought to further evaluate Abeta responses to LY-450139 in the guinea pig, a nontransgenic model that has an Abeta sequence identical to that of human. Male guinea pigs were treated with LY-450139 (0.2-60 mg/kg), and brain, cerebrospinal fluid, and plasma Abeta levels were characterized at 1, 3, 6, 9, and 14 h postdose. Low doses significantly elevated plasma Abeta levels at early time points, with return to baseline within hours. Higher doses inhibited Abeta levels in all compartments at early time points, but elevated plasma Abeta levels at later time points. To determine whether this phenomenon occurs under steady-state drug exposure, guinea pigs were implanted with subcutaneous minipumps delivering LY-450139 (0.3-30 mg/kg/day) for 5 days. Plasma Abeta was significantly inhibited at 10-30 mg/kg/day, but significantly elevated at 1 mg/kg/day. To further understand the mechanism of Abeta elevation by LY-450139, H4 cells overexpressing the Swedish mutant of amyloid-precursor protein and a mouse embryonic stem cell-derived neuronal cell line were studied. In both cellular models, elevated levels of secreted Abeta were observed at subefficacious concentrations, whereas dose-responsive inhibition was observed at higher concentrations. These results suggest that LY-450139 modulates the gamma-secretase complex, eliciting Abeta lowering at high concentrations but Abeta elevation at low concentrations.