Proteomic Analysis Reveals the Association between the Pathways of Glutathione and α-Linolenic Acid Metabolism and Lanthanum Accumulation in Tea Plants.

Lanthanum can affect the growth and development of the tea plant. Tieguanyin (TGY) and Shuixian (SX) cultivars of Camellia sinensis were selected to explore the mechanism underlying the accumulation of lanthanum (tea plants' most accumulated rare earth element) through proteomics. Roots and fresh leaves of TGY and SX with low- and high-accumulation potential for lanthanum, respectively, were studied; 845 differentially expressed proteins (DEPs) were identified. Gene ontology analysis showed that DEPs were involved in redox processes and related to molecular functions. Kyoto Encyclopedia of Genes and Genomes metabolic pathway analysis showed that DEPs were associated with glutathione (GSH) and α-linolenic acid metabolism, plant pathogen interaction, and oxidative phosphorylation. Thirty-seven proteins in the GSH metabolism pathway showed significant differences, wherein 18 GSH S-transferases showed differential expression patterns in the root system. Compared with the control, expression ratios of GST (TEA004130.1) and GST (TEA032216.1) in TGY leaves were 6.84 and 4.06, respectively, after lanthanum treatment; these were significantly higher than those in SX leaves. The LOX2.1 (TEA011765.1) and LOX2.1 (TEA011776.1) expression ratios in the α-linolenic acid metabolic pathway were 2.44 and 6.43, respectively, in TGY roots, which were significantly higher than those in SX roots. The synthesis of specific substances induces lanthanum-associated defense responses in TGY, which is of great significance for plant yield stability.

High Light Intensity Triggered Abscisic Acid Biosynthesis Mediates Anthocyanin Accumulation in Young Leaves of Tea Plant (Camellia sinensis).

There is increasing interest in the production and consumption of tea (Camellia sinensis L.) processed from purple-leaved cultivar due to their high anthocyanin content and health benefits. However, how and why seasonal changes affect anthocyanin accumulation in young tea leaves still remains obscured. In this study, anthocyanin and abscisic acid (ABA) contents in young leaves of Zifuxing 1 (ZFX1), a cultivar with new shoots turning to purple in Wuyi Mountain, a key tea production region in China, were monitored over four seasons. Young leaves produced in September were highly purplish, which was accompanied with higher anthocyanin and ABA contents. Among the environmental factors, the light intensity in particular was closely correlated with anthocyanin and ABA contents. A shade experiment also indicated that anthocyanin content significantly decreased after 168 h growth under 75% shade, but ABA treatment under the shade conditions sustained anthocyanin content. To confirm the involvement of ABA in the modulation of anthocyanin accumulation, anthocyanin, carotenoids, chlorophyll, ABA, jasmonic acid (JA), and salicylic acid (SA) in the young leaves of four cultivars, including ZFX1, Zijuan (ZJ), wherein leaves are completely purple, Rougui (RG) and Fudingdabaicha (FDDB) wherein leaves are green, were analyzed, and antioxidant activities of the leaf extracts were tested. Results showed that ABA, not other tested hormones, was significantly correlated with anthocyanin accumulation in the purple-leaved cultivars. Cultivars with higher anthocyanin contents exhibited higher antioxidant activities. Subsequently, ZFX1 plants were grown under full sun and treated with ABA and fluridone (Flu), an ABA inhibitor. ABA treatment elevated anthocyanin level but decreased chlorophyll contents. The reverse was true to those treated with Flu. To pursue a better understanding of ABA involvement in anthocyanin accumulation, RNA-Seq was used to analyze transcript differences among ABA- or Flu-treated and untreated ZFX1 plants. Results indicated that the differentially expressed genes in ABA or Flu treatment were mainly ABA signal sensing and metabolism-related genes, anthocyanin accumulation-related genes, light-responsive genes, and key regulatory MYB transcription factors. Taking all the results into account, a model for anthocyanin accumulation in ZFX1 cultivar was proposed: high light intensity caused reactive oxygen stress, which triggered the biosynthesis of ABA; ABA interactions with transcription factors, such as MYB-enhanced anthocyanin biosynthesis limited chlorophyll and carotenoid accumulation; and transport of anthocyanin to vacuoles resulting in the young leaves of ZFX1 with purplish coloration. Further research is warranted to test this model.

Interplay of Ecological Opportunities and Functional Traits Drives the Evolution and Diversification of Millettiod Legumes (Fabaceae).

Understanding the striking diversity of the angiosperms is a paramount issue in biology and of interest to biologists. The Millettiod legumes is one of the most hyper-diverse groups of the legume family, containing many economically important medicine, furniture and craft species. In the present study, we explore how the interplay of past climate change, ecological opportunities and functional traits' evolution may have triggered diversification of the Millettiod legumes. Using a comprehensive species-level phylogeny from three plastid markers, we estimate divergence times, infer habit shifts, test the phylogenetic and temporal diversification heterogeneity, and reconstruct ancestral biogeographical ranges. We found that three dramatic accumulations of the Millettiod legumes occurred during the Miocene. The rapid diversification of the Millettiod legumes in the Miocene was driven by ecological opportunities created by the emergence of new niches and range expansion. Additionally, habit shifts and the switch between biomes might have facilitated the rapid diversification of the Millettiod legumes. The Millettiod legumes provide an excellent case for supporting the idea that the interplay of functional traits, biomes, and climatic and geographic factors drives evolutionary success.

Combined analysis of lipidomics and transcriptomics revealed the key pathways and genes of lipids in light-sensitive albino tea plant (Camellia sinensis cv. Baijiguan).

Currently, the mechanism by which light-sensitive albino tea plants respond to light to regulate pigment synthesis has been only partially elucidated. However, few studies have focused on the role of lipid metabolism in the whitening of tea leaves. Therefore, in our study, the leaves of the Baijiguan (BJG) tea tree under shade and light restoration conditions were analyzed by a combination of lipidomics and transcriptomics. The leaf color of BJG was regulated by light intensity and responded to light changes in light by altering the contents and proportions of lipids. According to the correlation analysis, we found three key lipid components that were significantly associated with the chlorophyll SPAD value, namely, MGDG (36:6), DGDG (36:6) and DGDG (34:3). Further weighted gene coexpression network analysis (WGCNA) showed that HY5 TF and GLIP genes may be hub genes involved lipid regulation in albino tea leaves. Our results lay a foundation for further exploration of the color changes in albino tea leaves.

Willingness of Tea Farmers to Adopt Ecological Agriculture Techniques Based on the UTAUT Extended Model.

Ecological agricultural technology is the key method for making the transition from traditional agriculture to ecological agriculture, and is also the basic measure for promoting the transformation and upgrading of the tea industry and sustainable development. This study explores the influencing factors and mechanisms of tea farmers' adoption of ecological agricultural technology by using the extended model of the unified theory of technology adoption and use (UTAUT) based on perceived value. The analysis results, using the partial least squares structural equation model (PLS-SEM), show that: the positive impact of perceived value on willingness to use not only makes the explanatory power of the extended model greater than that of the original model but also expands the UTAUT model into a full mediating model, in which performance expectation has the greatest impact on behavioral intention through the implemented value. Effect expectation, social influence and factoring factors following, then the four intermediary paths have significant positive effects on behavioral intention. This study improves on the limitations of the UTAUT theoretical model through the theory of perceived value, and provides a reference for research on the same topic. At the same time, the government should provide tea farmers with enhanced subsidies, skills training and communication platforms.

Relationship between the Grade and the Characteristic Flavor of PCT (Panyong Congou Black Tea).

Panyong Congou black tea (PCT) is one of the most representative and historically famous Congou black teas in China and has been gaining more and more attention for its beneficial health properties. Currently, four grades of PCT are available, based on the raw leaf materials and consumer palatability. The chemical profiles distinguishing different grades of PCT are yet to be defined, nor has the relationship with grade been evaluated. In the present study, chemometric analysis showed that epigallocatechin (EGC), catechin (C), polyphenols, gallic acid (GA), and free amino acids are grade related bio-markers of PCT. These compounds are associated with the sweet and mellow aftertaste of PCT. A total of 34 volatile components were identified, of which the three component types with the highest relative percentages were alcohols (51.34-52.51%), ketones (27.31-30.28%), and aldehydes (12.70-13.18%). Additionally, our results revealed that sweet floral and fruity aromas were positively correlated with six volatile organic compounds (VOCs), 1-pentanol, propyl hexanoate, linalool, cyclohexanone, hexanal, and 2,5-dimethylpyrazine. Clear discrimination was achieved using orthogonal projections to latent structures discriminant analysis (OPLS-DA). The findings provide vital information on the characteristic flavor of each grade of PCT.

Erratum: Cardiomyocyte-Restricted Low Density Lipoprotein Receptor-Related Protein 6 (LRP6) Deletion Leads to Lethal Dilated Cardiomyopathy Partly Through Drp1 Signaling: Erratum.

[This corrects the article DOI: 10.7150/thno.22177.].

Widely Targeted Metabolomics Analysis Reveals the Differences of Nonvolatile Compounds in Oolong Tea in Different Production Areas.

The flavor differences in Oolong tea from different producing areas are caused by its complex differential compounds. In this study, representative samples of Oolong tea from four countries were collected, and their differential nonvolatile compounds were analyzed by a combination of widely targeted metabolomics, chemometrics, and quantitative taste evaluation. A total of 801 nonvolatile compounds were detected, which could be divided into 16 categories. We found that the difference in these compounds' content among Oolong teas from three producing areas in China was the largest. There were 370 differential compounds related to the producing areas of Oolong tea, which were mainly distributed in 67 Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways. In total, 81 differential nonvolatile compounds made important contributions to the taste differences in Oolong tea from different producing areas, among which the number of flavonoids was the largest. Finally, the characteristic compounds of Oolong tea in six producing areas were screened. This study comprehensively identifies the nonvolatile compounds of Oolong tea in different producing areas for the first time, which provides a basis for the analysis of flavor characteristics, quality directional control, and the identification and protection of geographical landmark agricultural products of Oolong tea from different producing areas.

Identification, Characterization and Expression Profiling of the RS Gene Family during the Withering Process of White Tea in the Tea Plant (Camellia sinensis) Reveal the Transcriptional Regulation of CsRS8.

Raffinose synthetase (RS) is a key enzyme in the process of raffinose (Raf) synthesis and is involved in plant development and stress responses through regulating Raf content. As a sweetener, Raf makes an important contribution to the sweet taste of white tea. However, studies on the identification, analysis and transcriptional regulation of CsRSs (Camellia sinensis RS genes) are still lacking. In this study, nine CsRSs were identified from the tea plant (Camellia sinensis) genome database. The CsRSs were classified into five groups in the phylogenetic tree. Expression level analysis showed that the CsRSs varied in different parts of the tea plant. Transcriptome data showed that CsRSs could respond to persistent drought and cold acclimation. Except for CsRS5 and CsRS9, the expression pattern of all CsRSs increased at 12 h and decreased at 30 h during the withering process of white tea, consistent with the change trend of the Raf content. Furthermore, combining yeast one-hybrid assays with expression analysis, we found that CsDBB could potentially regulate the expression of CsRS8. Our results provide a new perspective for further research into the characterization of CsRS genes and the formation of the white tea flavour.

[Analysis of clinical characteristics and prognostic risk factors in patients with Stanford type B aortic dissection complicated with hypertension].

OBJECTIVE: To investigate the clinical characteristics as well as short-term and long-term prognostic factors of patients with Stanford type B aortic dissection (TBAD) with hypertension. METHODS: Patients with TBAD who received thoracic endovascular aortic repair (TEVAR) admitted to Xiangyang Central Hospital from January 2014 to December 2018 were enrolled. The baseline data of patients admitted to the hospital were collected through the case management system, including gender, age, underlying diseases (hypertension, diabetes, coronary heart disease), smoking history, drinking history, duration of pain, vital signs at admission [heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP)], laboratory results [white blood cell count (WBC), platelet count (PLT), neutrophil/lymphocyte ratio (NLR), serum creatinine (SCr), C-reactive protein (CRP), D-dimer, ascending aorta diameter], etc. The clinical characteristics of TBAD patients with hypertension were analyzed. Logistic regression model and Cox proportional risk model were used to analyze the impact of hypertension on the short-term and long-term all-cause deaths after TEVAR in TBAD patients. RESULTS: Among 227 TBAD patients, 160 cases (70.5%) were complicated with hypertension, while 67 cases (29.5%) were not. The average age, the proportion of diabetes and coronary heart disease, and the level of SBP, DBP and SCr at admission of TBAD patients with hypertension were higher than those of TBAD without hypertension [age (years old): 53.1±11.9 vs. 42.8±14.1, combined with diabetes: 8.8% vs. 1.5%, combined with coronary heart disease: 6.3% vs. 0%, SBP (mmHg, 1 mmHg = 0.133 kPa): 147.9±18.1 vs. 136.9±15.2, DBP (mmHg): 93.9±11.9 vs. 89.1±13.8, SCr (µmol/L): 97.8±25.4 vs. 89.8±23.6, all P < 0.05]. The short-term mortality of TBAD with hypertension group was significantly higher than that of TBAD without hypertension group [6.3% (10/160) vs. 0% (0/67), χ2 = 4.386, P = 0.036]. 227 patients with TBAD were followed up for 3-66 months, with a median follow-up time of 25 months. There was no significant difference in long-term mortality between TBAD patients with and without hypertensive during discharge follow-up [13.1% (21/160) vs. 9.0% (6/67), χ2 = 0.784, P = 0.376]. Further multivariate Logistic regression analysis and Cox regression analysis did not indicate that hypertension was an independent risk factor for short-term and long-term death in TBAD patients [odds ratio (OR) and 95% confidence interval (95%CI) were 13.477 (0.541-330.215), 1.012 (0.990-1.035), both P > 0.05]. Age and HR were independent risk factors for the short-term mortality of TBAD patients [OR and 95%CI were 15.287 (1.051-226.415), 0.026 (0.002-0.840), both P < 0.05]. Age, PLT and D-dimer were independent risk factors for the long-term mortality of TBAD patients [OR and 95%CI were 1.808 (1.205-2.711), 0.555 (0.333-0.924), 1.482 (1.035-2.122), respectively, all P < 0.05]. CONCLUSIONS: The TBAD patients with hypertension have older age, high rates of diabetes or coronary heart disease. However, hypertension is not an independent risk factor for short-term and long-term mortality in TBAD patients.

Low-density lipoprotein receptor-related protein 6 regulates cardiomyocyte-derived paracrine signaling to ameliorate cardiac fibrosis.

Rationale: Maladaptive cardiac remodeling is a critical step in the progression of heart failure. Low-density lipoprotein receptor-related protein 6 (LRP6), a co-receptor of Wnt, has been implicated in cardiac protection. We aimed to study the role of cardiomyocyte-expressed LRP6 in cardiac remodeling under chronic pressure overload. Methods: Cardiac parameters were analyzed in inducible cardiac-specific LRP6 overexpressing and control mice subjected to transverse aortic constriction (TAC). Results: Cardiac LRP6 was increased at an early phase after TAC. Cardiomyocyte-specific LRP6 overexpression improved cardiac function and inhibited cardiac hypertrophy and fibrosis four weeks after TAC. The overexpression significantly inhibited ß-catenin activation, likely contributing to the inhibitory effect on cardiac hypertrophy after TAC. LRP6 overexpression reduced the expression and secretion of Wnt5a and Wnt11 by cardiomyocytes, and knockdown of Wnt5a and Wnt11 greatly inhibited cardiac fibrosis and dysfunction under pressure overload in vitro and in vivo. Cardiomyocyte-expressed LRP6 interacted with cathepsin D (CTSD, a protease) and promoted the degradation of Wnt5a and Wnt11, inhibiting cardiac fibrosis and dysfunction induced by TAC. The protease inhibitor leupeptin attenuated the interaction between LRP6 and CTSD, enhanced the expression of Wnt5a and Wnt11, and deteriorated cardiac function and fibrosis in cardiomyocyte-specific LRP6-overexpressing mice under pressure overload. Mutants from human patients, P1427Q of LRP6 and G316R of CTSD significantly inhibited the interaction between LRP6 and CTSD and increased Wnt5a and Wnt11 expression. Conclusion: Cardiomyocyte-expressed LRP6 promoted the degradation of Wnt5a and Wnt11 by regulating CTSD and inhibited cardiac fibrosis under pressure overload. Our study demonstrated a novel role of LRP6 as an anti-fibrosis regulator.

[Evaluation of Red Blood Cell Indices and Parameter Formulas in Differential Diagnosis of Thalassemia Trait and Iron Deficiency Anemia for Children in Shenzhen Area of Guangdong Province in China].

OBJECTIVE: To evaluate the efficiency of red blood cell indices and fomulas for the differential diagnosis of the thalassemia trait (TT) and iron deficiency anemia (IDA) for children in Shenzhen area of Guangdong Province in China. METHODS: A total of 849 child patients from Shenzhen were enrolled, including 536 cases of TT and 313 cases of IDA. The sensitivity (SEN), specificity (SPE), positive predictive values (PPV), negative predictive value (NPV), and Youden's indices (YI) were analyzed using five red blood cell indices ï¼»including red blood cell count, average red blood cell volune(MCV), average amount of red blood cell hemoglobin(CMH), red blood hemoglobin cancentration(MCHC), red blood cell distribution width(RDW)ï¼½ and 10 red blood cell paramter formulas including Mentzer, Green and King, Srivastava, Ricerca, RDWI, Sirdah, Huber-Herklotz, Ehsani, Shine and Lal, and England and Fraser. Receiver operating characteristic (ROC) curve was drawn. RESULTS: Green and King was the most reliable index, as it had the highest YI (63.7%) and area under ROC curve (AUC) (0.875), the SEN and SPE was 82.5% and 81.2%. The YI, SEN, SPE, and AUC for RDWI were 62.8%, 79.1%, 83.7%, and 0.870, respectively. CONCLUSION: The formulas of Green and King and RDWI can be used for the differential diagnosis of TT and IDA, suitable for chidren in Shenzhen, China.

Low density lipoprotein receptor related protein 6 (LRP6) protects heart against oxidative stress by the crosstalk of HSF1 and GSK3ß.

Low density lipoprotein receptor-related protein 6 (LRP6), a Wnt co-receptor, induces multiple functions in various organs. We recently reported cardiac specific LRP6 deficiency caused cardiac dysfunction in mice. Whether cardiomyocyte-expressed LRP6 protects hearts against ischemic stress is largely unknown. Here, we investigated the effects of cardiac LRP6 in response to ischemic reperfusion (I/R) injury. Tamoxifen inducible cardiac specific LRP6 overexpression mice were generated to build I/R model by occlusion of the left anterior descending (LAD) coronary artery for 40 min and subsequent release of specific time. Cardiac specific LRP6 overexpression significantly ameliorated myocardial I/R injury as characterized by the improved cardiac function, strain pattern and infarct area at 24 h after reperfusion. I/R induced-apoptosis and endoplasmic reticulum (ER) stress were greatly inhibited by LRP6 overexpression in cardiomyocytes. LRP6 overexpression enhanced the expression of heat shock transcription factor-1(HSF1) and heat shock proteins (HSPs), the level of p-glycogen synthase kinase 3ß(GSK3ß)(S9) and p-AMPK under I/R. HSF1 inhibitor deteriorated the apoptosis and decreased p-GSK3ß(S9) level in LRP6 overexpressed -cardiomyocytes treated with H2O2. Si-HSF1 or overexpression of active GSK3ß significantly attenuated the increased expression of HSF1 and p-AMPK, and the inhibition of apoptosis and ER stress induced by LRP6 overexpression in H2O2-treated cardiomyocytes. AMPK inhibitor suppressed the increase in p-GSK3ß (S9) level but didn't alter HSF1 nucleus expression in LRP6 overexpressed-cardiomyocytes treated with H2O2. Active GSK3ß, but not AMPK inhibitor, attenuated the inhibition of ubiquitination of HSF1 induced by LRP6-overexpressed-cardiomyocytes treated with H2O2. LRP6 overexpression increased interaction of HSF1 and GSK3ß which may be involved in the reciprocal regulation under oxidative stress. In conclusion, cardiac LRP6 overexpression significantly inhibits cardiomyocyte apoptosis and ameliorates myocardial I/R injury by the crosstalk of HSF1 and GSK3ß signaling.

Cardiac-specific LRP6 knockout induces lipid accumulation through Drp1/CPT1b pathway in adult mice.

We recently reported low-density lipoprotein receptor-related protein 6 (LRP6) decreased in dilated cardiomyopathy hearts, and cardiac-specific knockout mice displayed lethal heart failure through activation of dynamin-related protein 1 (Drp1). We also observed lipid accumulation in LRP6 deficiency hearts, but the detailed molecular mechanisms are unclear. Here, we detected fatty acids components in LRP6 deficiency hearts and explored the potential molecular mechanisms. Fatty acid analysis by GC-FID/MS revealed cardiac-specific LRP6 knockout induced the higher level of total fatty acids and some medium-long-chain fatty acids (C16:0, C18:1n9 and C18:2n6) than in control hearts. Carnitine palmitoyltransferase 1b (CPT1b), a rate-limiting enzyme of mitochondrial ß-oxidation in adult heart, was sharply decreased in LRP6 deficiency hearts, coincident with the activation of Drp1. Drp1 inhibitor greatly improved cardiac dysfunction and attenuated the increase in total fatty acids and fatty acids C16:0, C18:1n9 in LRP6 deficiency hearts. It also greatly inhibited the decrease in the cardiac expression of CPT1b and the transcriptional factors CCCTC-binding factor (CTCF) and c-Myc induced by cardiac-specific LRP6 knockout in mice. C-Myc but not CTCF was identified to regulate CPT1b expression and lipid accumulation in cardiomyocytes in vitro. The present study indicated cardiac-specific LRP6 knockout induced lipid accumulation by Drp1/CPT1b pathway in adult mice, and c-Myc is involved in the process. It suggests that LRP6 regulates fatty acid metabolism in adult heart.

Lipoprotein receptor-related protein 6 is required to maintain intercalated disk integrity.

The intercalated disk (ID), a highly organized adhesion structure connecting neighboring cardiomyocytes, fulfills mechanical and electrical signaling communication to ensure normal heart function. Lipoprotein receptor-related protein 6 (LRP6) is a co-receptor inducing canonical Wnt/ß-catenin signaling. It was recently reported that LRP6 deficiency in cardiomyocytes predisposes to arrhythmia independent of Wnt signaling. However, whether LRP6 directly regulates the structure of IDs requires further investigation. The aim of the present study was to explore the role of LRP6 in IDs and the potential underlying mechanisms by inducible cardiac-specific LRP6 knockout mice. The results revealed that LRP6 was predominately expressed in the cell membrane, including the IDs of cardiomyocytes. Tamoxifen-inducible cardiac-specific LRP6 knockout mice displayed overt cardiac dysfunction and disruption of ID structure. Further analysis revealed that cardiac LRP6 deficiency induced the imbalance of ID component proteins, characterized by the sharply decreased expression of connexin 43 (Cx43) and the significantly increased expression of N-cadherin, desmoplakin and γ-catenin in tissue lysates or membrane fraction from the left ventricle. STRING database analysis indicated that ß-catenin, but no other ID-associated proteins, interacted with LRP6. Our immunoprecipitation analysis demonstrated that LRP6 strongly interacted with Cx43, N-cadherin and γ-catenin, and weakly interacted with ß-catenin, whereas there was no association with desmoplakin. In response to LRP6 deficiency, the recruitment of ß- or γ-catenin to N-cadherin was increased, but they displayed little interaction with Cx43. In conclusion, LRP6 is required to maintain the integrity of ID structure and the balance of ID proteins, and the interaction between LRP6 and Cx43, N-cadherin and γ-catenin may be involved in this process.

Strong impact of sulfotransferases on DNA adduct formation by 4-aminobiphenyl in bladder and liver in mice.

Bladder cancer risk is 3-4 times higher in men than women, but the reason is poorly understood. In mice, male bladder is also more susceptible than female bladder to 4-aminobiphenyl (ABP), a major human bladder carcinogen; however, female liver is more susceptible than male liver to ABP. We investigated the role of sulfotransferase (Sult) in gender-related bladder and liver susceptibility to ABP. Sulfation reactions of aromatic amine bladder carcinogens catalyzed by Sult may generate highly unstable and toxic metabolites. Therefore, liver Sult may decrease bladder exposure to carcinogens by promoting their toxic reactions in the liver. Notably, the expression of several liver Sults is suppressed by androgen in male mice. Here, we show that two Sults are critical for gender-related bladder susceptibility to ABP in mice. We measured tissue level of N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dG-C8-ABP), a principal ABP-DNA adduct, as readout of tissue susceptibility to ABP. We identified Sutl1a1 and to a lesser extent Sult1d1 as Sults that promote dG-C8-ABP formation in hepatic cells. In mice, gender gap in bladder susceptibility to ABP was narrowed by knocking out Sult1a1 and was almost totally eliminated by knocking out both Sutl1a1 and Sult1d1. This was accompanied by dramatic decrease in ABP genotoxicity in the liver (>97%). These results show the strong impact of the Sults on bladder and liver susceptibility to a human carcinogen. Because liver expression of both Sult1a1 and Sutl1d1 is suppressed by androgen in male mice, our results suggest that androgen renders bladder more exposed to ABP in male mice by suppressing Sult-mediated ABP metabolism in liver, which increases bladder delivery of carcinogenic metabolites.

Mechanical stresses induce paracrine ß-2 microglobulin from cardiomyocytes to activate cardiac fibroblasts through epidermal growth factor receptor.

By employing a proteomic analysis on supernatant of mechanically stretched cardiomyocytes, we found that stretch induced a significantly high level of ß-2 microglobulin (ß2M), a non-glycosylated protein, which is related to inflammatory diseases but rarely known in cardiovascular diseases. The present data showed that serum ß2M level was increased in patients with hypertension and further increased in patients with chronic heart failure (HF) as compared with control group, and the high level of serum ß2M level correlated to cardiac dysfunction in these patients. In pressure overload mice model by transverse aortic constriction (TAC), ß2M levels in serum and heart tissue increased progressively in a time-dependent manner. Exogenous ß2M showed pro-fibrotic effects in cultured cardiac fibroblasts but few effects in cardiomyocytes. Adeno-associated virus 9 (AAV9)-mediated knockdown of ß2M significantly reduced cardiac ß2M level and inhibited myocardial fibrosis and cardiac dysfunction but not cardiac hypertrophy at 4 weeks after TAC. In vitro, mechanical stretch induced the rapid secretion of ß2M mainly from cardiomyocytes by activation of extracellular-regulated protein kinase (ERK). Conditional medium (CM) from mechanically stretched cardiomyocytes activated cultured cardiac fibroblasts, and the effect was partly abolished by CM from ß2M-knockdown cardiomyocytes. In vivo, knockdown of ß2M inhibited the increase in phosphorylation of epidermal growth factor receptor (EGFR) induced by TAC. In cultured cardiac fibroblasts, inhibition of EGFR significantly attenuated the ß2M-induced the activation of EGFR and pro-fibrotic responses. The present study suggests that ß2M is a paracrine pro-fibrotic mediator and associated with cardiac dysfunction in response to pressure overload.

Up-regulated expression of miR-155 in human colonic cancer.

OBJECTIVE: The aim of this study is to investigate the expression of miR-155 in colonic cancer tissue and to assess the potential predictive value of miR-155 in colonic cancer patients. MATERIALS AND METHODS: From March to September of 2011, we included 57 patients with primary colonic cancer who underwent curative surgical resection. Total RNAs were extracted from colonic cancer tissues and adjacent normal tissues. Then the expression of miR-155 in colonic cancer and paracancerous tissues was investigated using real time quantitative reverse transcription-polymerase chain reaction. And the relationship between miR-155 expression level and the clinical, pathological parameters of colonic cancer was analyzed. RESULTS: The relative expression level of miR-155 in colonic cancer tissues was significantly higher than that in paracancerous tissues and related to tumor-node-metastasis staging, tumor invasion, metastasis, and differentiation. CONCLUSION: The expression of miR-155 is up-regulated in colonic cancer tissues. MiR-155 may acts as proto-oncogenes involved in carcinogenesis, development, and invasion of colon cancer making it a potential target for gene therapy of colon cancer.

Knockdown of LRP6 activates Drp1 to inhibit survival of cardiomyocytes during glucose deprivation.

Lipoprotein receptor-related protein 6 (LRP6) binds to Wnt ligands to transduce signal by stabilization of ß-catenin, which has been involved in the regulation of embryonic development and metabolism et al. Here, we observed LRP6 decreased in human hearts with dilated cardiomyopathy (DCM), and it also decreased in cultured cardiomyocytes under glucose- deprivation (GD). Knockdown of LRP6 greatly inhibited cell viability in cardiomyocytes under GD, but it didn't induce the effect in cardiomyocytes at baseline. Overexpression of LRP6 increased the cell viability in GD-cardiomyocytes. To explore potential molecular mechanisms, we detected the phosphorylation of dynamin-related protein 1(Drp1) and active ß-catenin in cardiomyocytes under GD. Knockdown of LRP6 enhanced p-Drp1(S616) level while it didn't alter the p-Drp1(S637) and active ß-catenin level in GD-cardiomyocytes. Drp1 inhibitor significantly suppressed the increase in p-Drp1 at S616 and improved the cell viability in GD-cardiomyocytes with knockdown of LRP6. Further analysis showed that knockdown of LRP6 also increased the phosphorylation of mammalian target of rapamycin (mTOR), and Drp1 inhibitor greatly inhibited the increase in p-mTOR level in GD-cardiomyocytes. The present study indicated that knockdown of LRP6 inhibited the cell viability by activation of Drp1 in GD-cardiomyocytes, and the phosphorylation of mTOR may be involved in the process. It suggests that LRP6 can prevent cardiomyocytes from death in nutrition-deprived condition.