Long noncoding RNA00324 is involved in the inflammation of rheumatoid arthritis by targeting miR-10a-5p via the NF-κB pathway.

BACKGROUND: Altered expressions of genes in immune cells and synovial tissues are involved in the pathology of rheumatoid arthritis (RA). Long noncoding RNAs act as competing endogenous RNAs and can cause immune disorders. The goal of this study was to reveal the association between noncoding RNA linc00324 and RA, and a plausible action mechanism was proposed. METHODS: RT-qPCR was used to evaluate the expression of linc00324 in peripheral blood mononuclear cells isolated from 50 RA patients and 50 healthy controls, and the correlations between linc00324 level and the clinical indicators were analyzed. Flow cytometry was used to characterize CD4+ T cells. The effects of linc00324 on cytokine production and cell proliferation of CD4+ T cells were evaluated by ELISA assay and Western blot. The interaction between linc00324 and miR-10a-5p was investigated by RNA immunoprecipitation and dual-luciferase assays. RESULTS: The linc00324 expression was significantly enhanced in RA patients, and linc00324 expression was positively correlated with rheumatoid factor and CD4+ T cells. Overexpression of linc00324 promoted CD4+ T cells proliferation, and enhanced chemokine MIP-1α secretion and NF-κB phosphorylation level, whereas knockout of linc00324 blocked CD4+ T cell proliferation and NF-κB phosphorylation. Overexpression of miR-10a-5p led to the decrease of CD4+ T cells proliferation and NF-κB phosphorylation, and reversed the effects of linc00324 on cell proliferation and NF-κB activity. CONCLUSION: Linc00324 was upregulated in RA and may exaggerate inflammation by targeting miR-10a-5p through NF-κB signaling pathway.

The effect of dietary awareness on household food waste.

How to reduce household food waste has emerged as an important issue worldwide. Considering the potential endogeneity issue, the present study utilizes the treatment effects model to analyse the effect of dietary awareness on household food waste using data from China. The results showed that improving dietary awareness could significantly increase household food waste overall. However, this impact is heterogeneous among households of different characteristics. Improving dietary awareness leads to more food waste for households with young and old food decision-makers, but contributes to less food waste for households of middle-aged makers. Also, the positive effect of dietary awareness on household food waste weakens as income increases. These findings propose a new perspective to understand the heterogeneity in household food waste in the context of dietary awareness promotion.

The Direct and Structure Effect of Income on Nutrition Demand of Chinese Rural Residents.

Although a significant body of literature has analyzed the effect of income-mediated policies on nutrition, research on how income affect nutrition consumption is scant. This paper contributes to the literature by decomposing the overall income effect on rural residents' calorie intake into the direct income effect and the structure effect by building a simple theoretical model and conducting related empirical research with an instrumental variable (IV) approach. Using nationally representative data from China, we find that the structure effect of income, represented by fat share growth induced by income, occupies a considerable proportion (38.03%) of overall income effect. Additionally, we provide evidence of an asymmetric distributional effect of income on calorie intake. In particular, the structure effect of income substantially accounts for a larger proportion in the higher quantiles of the calorie intake distribution. Our findings help better evaluate the effectiveness of the income-mediated policies from quantity and structure perspectives in a comprehensive framework.

cDNA cloning of a novel lectin that induce cell apoptosis from Artocarpus hypargyreus.

We isolated a novel lectin (AHL) from Artocarpus hypargyreusHance and showed its immunomodulatory activities. In this study, the amino acid sequence of AHL was determined by cDNA sequencing. AHL cDNA (875bp) contains a 456-bp open reading frame (ORF), which encodes a protein with 151 amino acids. AHL is a new member of jacalin-related lectin family (JRLs), which share high sequence similarities to KM+ and Morniga M, and contain the conserved carbohydrate binding domains. The antitumor activity of AHL was also explored using Jurkat T cell lines. AHL exhibits a strong binding affinity to cell membrane, which can be effectively inhibited by methyl-α-D-galactose. AHL inhibits cell proliferation in a time- and dose-dependent manner through apoptosis, evidenced by morphological changes, phosphatidylserine externalization, poly ADP-ribose polymerase (PARP) cleavage, Bad and Bax up-regulation, and caspase-3 activation. We further showed that the activation of ERK and p38 signaling pathways is involved for the pro-apoptotic effect of AHL.

Long intergenic non-protein coding RNA 324 prevents breast cancer progression by modulating miR-10b-5p.

Mounting evidence suggests that long noncoding RNAs serve as specific biomarkers and potent modulators of multiple cancers. Long intergenic non-protein coding RNA 324 (LINC00324) is ubiquitously expressed in various tissues, including breast cancer. The biological function of LINC00324 in the development and progression of breast cancer remains unknown. Here, we fully elucidate the relation between LINC00324 expression and breast cancer, and suggest a potential mechanism of action. We found that decreased expression of LINC00324 was dramatically correlated with malignancy of breast cancer, both in breast cancer tissues and in cell lines. Overexpression of LINC00324 in MDA-MB-231 cells resulted in a decrease in cell proliferation, invasion, and migration, while increasing cells apoptosis. On the other hand, loss-of-function experiments indicated that deficiency of LINC00324 promoted malignant phenotypes in breast cancer cells. Mechanically, we found that LINC00324 is mainly distributed in the cytoplasm, fostering the expression of E-cadherin by sponging miR-10b-5p. Taken together, these findings suggest that LINC00324 plays a critical role in breast cancer progression by directly interacting with miR-10b-5p. LINC00324 can thus potentially act as an early diagnostic marker and a novel therapeutic agent for breast cancer.

[Correlation analysis between LINC00324 and immunophenotype in peripheral blood leukocytes in patients with acute myeloid leukemia].

Objective To investigate the relationship between long-chain intergenic non-coding RNA324 (LINC00324) and immunophenotype in peripheral blood leukocytes of acute myeloid leukemia (AML) patients. Methods Real-time quantitative PCR and bioinformatics databases were utilized to analyze the expression level of LINC00324 in peripheral blood leukocytes and cell lines KG-1, THP-1 and U937 in AML patients. The relationships of the expression level of LINC00324 with the red blood cell and platelet count, the expression levels of LINC00324 and immunophenotypes in 40 AML patients were analyzed by Person correlation analysis. The immunophenotypes included CD14, CD68, CD64, CD11b, CD4, CD45, CD33, HLA-DR, CD163, CD2, CD58, CD117, CD43, CD34, CD99, CD8, CD38, CD10, CD13, CD56, CD7, TdT, CD235a, CD138, CD61, MPO and CD19. Simultaneously, the cBioPortal database datasets (TCGA, NEJM 2013) were used to analyze the clinical characteristics of 173 AML patients, and to analyze the correlations between the expression level of LINC00324 and the peripheral blood blast percentage and white blood cell count in tumor samples. Results The expression of LINC00324 in peripheral blood leukocytes of AML patients was down-regulated, and its expression level was significantly correlated with immunophenotype CD33, red blood cell and platelet count. Analysis of bioinformatics database showed that LINC00324 was under-expressed in myeloid leukemia cell lines. The expression of LINC00324 in AML patients was associated with multiple immunophenotypes such as CD33, CD117, CD11b, CD14 and CD64 and was negatively correlated with peripheral blood blast percentage and white blood cell count. Conclusion LINC00324 may be involved in regulating the differentiation, development and function of immune cells, which providing a new strategy for the development of targeted drugs or treatment of AML.

Medical system and nutrition improvement for the rural elderly.

BACKGROUND: Insufficient nutrition intake has negatively influenced the health of the elderly in rural China where the problem of population aging is serious. The present study aims to explore whether the medical system, called the New Rural Cooperative Medical System (NRCMS), can improve the rural elderly's nutrition intake and the mechanism behind it. METHODS: The difference in differences (DID) model and the propensity score matching-difference in differences (PSM-DID) model are both performed to investigate the impact of the medical system on nutrition improvement for the rural elderly. Two thousand seven hundred eighty rural elderly samples tracked in 2000 and 2006 from the China Health and Nutrition Survey are analyzed. Indices for the elderly's nutrition intake includes daily average intake of energy, fat, protein, and carbohydrate. RESULTS: The results show that participation in the NRCMS can significantly increase the rural elderly's total energy intake, carbohydrate intake, and protein intake by 206.688 kcal, 36.379 g, and 6.979 g, respectively. A more significant impact of the NRCMS on nutrition intake is observed in the central and near-western where economic development is lagging behind. Also, compared to people of 18-60 age group, such impact is statistically more significant in the elderly for the carbohydrate intake. CONCLUSIONS: The NRCMS can improve the rural elderly's nutrition intake in China. As the population ages rapidly in rural China, the present study provides recommendations on how to improve nutrition and health status of the elderly from the aspect of the medical system.

Phosphatase PPM1L Prevents Excessive Inflammatory Responses and Cardiac Dysfunction after Myocardial Infarction by Inhibiting IKKß Activation.

Although the inflammatory response triggered by damage-associated molecular patterns (DAMPs) in the infarcted cardiac tissues after acute myocardial infarction (MI) contributes to cardiac repair, the unrestrained inflammation induces excessive matrix degradation and myocardial fibrosis, leading to the development of adverse remodeling and cardiac dysfunction, although the molecular mechanisms that fine tune inflammation post-MI need to be fully elucidated. Protein phosphatase Mg2+/Mn2+-dependent 1L (PPM1L) is a member of the serine/threonine phosphatase family. It is originally identified as a negative regulator of stress-activated protein kinase signaling and involved in the regulation of ceramide trafficking from the endoplasmic reticulum to Golgi apparatus. However, the role of PPM1L in MI remains unknown. In this study, we found that PPM1L transgenic mice exhibited reduced infarct size, attenuated myocardial fibrosis, and improved cardiac function. PPM1L transgenic mice showed significantly lower levels of inflammatory cytokines, including IL-1ß, IL-6, TNF-α, and IL-12, in myocardial tissue. In response to DAMPs, such as HMGB1 or HSP60, released in myocardial tissue after MI, macrophages from PPM1L transgenic mice consistently produced fewer inflammatory cytokines. PPM1L-silenced macrophages showed higher levels of inflammatory cytokine production induced by DAMPs. Mechanically, PPM1L overexpression selectively inhibited the activation of NF-κB signaling in myocardial tissue post-MI and DAMP-triggered macrophages. PPM1L directly bound IKKß and then inhibited its phosphorylation and activation, leading to impaired NF-κB signaling activation and suppressed inflammatory cytokine production. Thus, our data demonstrate that PPM1L prevents excessive inflammation and cardiac dysfunction after MI, which sheds new light on the protective regulatory mechanism underlying MI.

Purification and characterization of a novel immunomodulatory lectin from Artocarpus hypargyreus Hance.

Lectins are proteins/glycoproteins of non-immune origin, which interact specifically and non-covalently with the carbohydrate moieties of respective receptors on the cell surface. In this study, a novel 65.2-kDa tetrameric lectin (AHL) was purified from Artocarpus hypargyreus Hance (A. hypargyreus) by affinity chromatography on a galactose-sepharose column. It is a glycoprotein with carbohydrate content of 6.91%. Its maximum haemagglutinating activity was maintained after incubation at a temperature range of 20-40 °C and pH range of 5.0-9.0. AHL-induced haemagglutination of erythrocytes was inhibited strongly by carbohydrates, such as methyl-galactose, methyl-mannose, and N-acetyl-d-galactosamine, indicating the existence of more than one carbohydrate binding sites in the AHL molecule. The AHL activity was gradually lost in the presence of urea and completely lost when being treated with ethylenediaminetetraacetic acid (EDTA). The immunomodulatory activity of AHL was assessed using human peripheral lymphocytes and rat peritoneal macrophages. AHL triggered proliferation and activation of human T lymphocytes and induced the release of Th1 cytokines, including IFN-γ, TNF-α and IL-6. Furthermore, AHL significantly stimulated the production of nitric oxide (NO) and pro-inflammatory cytokines, including TNF-α and IL-12, in rat peritoneal macrophages. However, AHL did not enhance proliferation of B cell-enriched rat splenocytes. Taken together, in this study, a novel immunomodulatory lectin was purified from A. hypargyreus and was found to be capable of inducing a Th1-type immune response, and thus, it may have potential immunoregulatory application in response to infections, immune diseases and cancer.

Overweight and obesity standards and subjective well-being: Evidence from China.

The adult BMI cutoffs for overweight and obesity standards set by the Working Group on Obesity in China are exerting growing influence over daily life. Using the regression discontinuity design method, this paper confirms the existence of a statistically significant discontinuity in subjective well-being at the overweight and obesity cutoffs, respectively. The overweight standard causes a significant decrease in subjective well-being (SWB) by approximate 0.10 units, and the obesity standard by 0.14 units, both sizable. Thus the standard setting has profound social, economic and welfare impacts beyond the health scope.

Mitogenic activity of Artocarpus lingnanensis lectin and its apoptosis induction in Jurkat T cells.

Lectins are a class of carbohydrate-binding proteins or glycoproteins and used in the purification and characterization of glycoproteins according to their specificity to carbohydrates. In the present study, the mitogenic activity of Artocarpus lingnanensis lectin (ALL) and its apoptosis induction in Jurkat T cells were explored. MTT assay revealed strong mitogenic potential of ALL. Meanwhile, the anti-cancer activity of ALL was also explored using the human leukemic Jurkat T cell line. ALL exhibited strong binding affinity (97%) to the cell membrane, which could be effectively inhibited by N-acetyl-D-galactosaminide (NAD). ALL induced time- and dose-dependent growth inhibition in Jurkat T cells. ALL could induce morphologic change and increase the hypodiploid cell population with the decreased population of S and G2/M phases. The induction of phosphatidylserine externalization and PARP cleavage further confirmed its apoptosis-inducing activity due to the activation of caspase-8 and -9. The inhibition of caspase-9 but not caspase-8 could rescue ALL-induced apoptotic cells. Further studies showed that ALL enhanced the cleavage of Bid, the release of cytochrome C, the depolarization of mitochondria and the activation of caspase-3. ALL downregulated the expression of Bcl-xl and Bcl-2 without impact on Bax and Bad. In addition, the activation of p38/JNK MAPK signaling pathways was observed to be a requisite for ALL apoptotic activity. In contrast, ALL could not induce apoptosis of normal T cells. These findings present the differential effect of ALL on Jurkat and normal T lymphocytes, suggesting its therapeutic value in leukemia.

Cloning of a novel lectin from Artocarpus lingnanensis that induces apoptosis in human B-lymphoma cells.

We isolated a novel lectin (Artocarpus nitidus subsp. lingnanensis lectin, ALL) from Artocarpus nitidus subsp. lingnanensis and showed its mitogenic activities. In this study, we determined the amino acid sequence of ALL by cDNA sequencing. ALL cDNA (933 bp) contains a 657-bp open reading frame (ORF), which encodes a protein with 218 amino acids. ALL shares high sequence similarities with Jacalin and Morniga G and belongs to jacalin-related lectin family. We also examined the antitumor activity of ALL using Raji, a human B-lymphoma cell line. ALL exhibits a strong binding affinity to cell membrane, which can be effectively inhibited by N-acetyl-D-galactosamine (GalNAc). ALL inhibits Raji cell proliferation in a time- and dose-dependent manner through apoptosis, evidenced by morphological changes, phosphatidylserine externalization, poly ADP-ribose polymerase (PARP) cleavage, Bcl-2 down-regulation, and caspase-3 activation. We further showed that the activation of p38 mitogen-activated protein kinase (MAPK) signaling pathways is required for the pro-apoptotic activity of ALL.

Eating out and getting fat? A comparative study between urban and rural China.

In parallel with the increased prevalence of overweight and obesity, the rate of food away from home (FAFH) consumption in China has increased notably in recent years. Under the long-term urban-rural dual structure in China, the purpose of this study was to investigate the impact of FAFH consumption on body mass index (BMI) by a comparative study between rural and urban areas, using 26,244 subjects from the 2004-2011 China Health and Nutrition Survey. The results indicated that urban residents have a higher rate of FAFH consumption than rural residents with the difference narrowing over time. The empirical results illustrated that the frequency of meals consumed away from home had a significantly positive effect on BMI in urban China, whereas no significant association was observed in rural China. The urban-rural difference resulted from different levels of surplus energy, which was mainly due to the different labor intensity among rural and urban residents.

The impact of changes in dietary knowledge on adult overweight and obesity in China.

Overweight and obesity are rapidly growing threats in China. Improvement in dietary knowledge can potentially prevent overweight and obesity, conditions which are receiving substantial attention from international organizations and governments. The purpose of this study was to investigate the impact of changes in dietary knowledge on adult overweight and obesity, using a balanced panel data consisting of 10,401 samples from the 2006, 2009, and 2011 iterations of the China Health and Nutrition Survey. Results indicate that overweight and obesity are becoming increasingly problematic in China, and the level of dietary knowledge among Chinese adults needs improvement. Moreover, the empirical results indicate that changes in dietary knowledge among adults has no significant influence on adult overweight and obesity, a likely result of lacking systematic dietary knowledge and having inadequate guidance on overweight/obesity-related behaviors.

A novel lectin from Artocarpus lingnanensis induces proliferation and Th1/Th2 cytokine secretion through CD45 signaling pathway in human T lymphocytes.

Lectins are carbohydrate-binding proteins and have been used for purification and characterization of glycoproteins. In this study, a novel 58.9-kDa tetrameric lectin from Artocarpus lingnanensis seeds was purified, characterized, and its mitogenic potential was evaluated. The hemagglutination inhibition assay indicated that Artocarpus lingnanensis lectin (ALL) showed specificity toward galactose. ALL was effectively purified in a single-step using affinity chromatography on a galactose-Sepharose column. ALL showed pH optima between 5.0 and 9.0, and optimal temperature between 20 and 40 °C. ALL triggered proliferation and activation of human T lymphocytes (e.g., CD4+ T lymphocytes). Flow cytometry and laser scanning confocal microscopy revealed binding of ALL to T cells and colocalized with CD45. Affinity chromatography and Western blot suggested that CD45 isolated from human T cell membrane fraction may be the major receptor of ALL. CD45 blocking antibody attenuated the binding and proliferation of T cells induced by ALL. CD45-PTPase inhibitor dephostatin reduced ALL-induced T cells proliferation and expression of CD25 and pZAP-70. Furthermore, secretion of ALL-induced Th1/Th2 cytokines was blocked with dephostatin. Also, dephostatin inhibited phosphorylation of ALL-mediated activation of ERK and p38MAPK. This study demonstrates the involvement of CD45-mediated signaling in ALL-induced T lymphocyte proliferation and Th1/Th2 cytokine secretion through activation of p38 and ERK.

Sensitization of cervical carcinoma cells to paclitaxel by an IPP5 active mutant.

Paclitaxel is one of the best anticancer agents that has been isolated from plants, but its major disadvantage is its dose-limiting toxicity. In this study, we obtained evidence that the active mutant IPP5 (8-60hIPP5m), the latest member of the inhibitory molecules for protein phosphatase 1, sensitizes human cervix carcinoma cells HeLa more efficiently to the therapeutic effects of paclitaxel. The combination of 8-60hIPP5m with paclitaxel augmented anticancer effects as compared to paclitaxel alone as evidenced by reduced DNA synthesis and increased cytotoxicity in HeLa cells. Furthermore, our results revealed that 8-60hIPP5m enhances paclitaxel- induced G2/M arrest and apoptosis, and augments paclitaxel-induced activation of caspases and release of cytochrome C. Evaluation of signaling pathways indicated that this synergism was in part related to down- regulation of NF-?B activation and serine/threonine kinase Akt pathways. We noted that 8-60hIPP5m down- regulated the paclitaxel-induced NF-?B activation, I?Bα degradation, PI3-K activity and phosphorylation of the serine/threonine kinase Akt, a survival signal which in many instances is regulated by NF-?B. Together, our observations indicate that paclitaxel in combination with 8-60hIPP5m may provide a therapeutic advantage for the treatment of human cervical carcinoma.

8-60hIPP5(m)-induced G2/M cell cycle arrest involves activation of ATM/p53/p21(cip1/waf1) pathways and delayed cyclin B1 nuclear translocation.

Protein phosphatase 1 (PP1) is a major serine/threonine phosphatase that controls gene expression and cell cycle progression. The active mutant IPP5 (8-60hIPP5(m)), the latest member of the inhibitory molecules for PP1, has been shown to inhibit the growth of human cervix carcinoma cells (HeLa). In order to elucidate the underlying mechanisms, the present study assessed overexpression of 8-60hIPP5(m) in HeLa cells. Flow cytometric and biochemical analyses showed that overexpression of 8-60hIPP5(m) induced G2/M-phase arrest, which was accompanied by the upregulation of cyclin B1 and phosphorylation of G2/M-phase proteins ATM, p53, p21(cip1/waf1) and Cdc2, suggesting that 8-60hIPP5(m) induces G2/M arrest through activation of the ATM/p53/p21(cip1/waf1)/Cdc2/ cyclin B1 pathways. We further showed that overexpression of 8-60hIPP5(m) led to delayed nuclear translocation of cyclin B1. 8-60hIPP5(m) also could translocate to the nucleus in G2/M phase and interact with pp1α and Cdc2 as demonstrated by co-precipitation assay. Taken together, our data demonstrate a novel role for 8-60hIPP5(m) in regulation of cell cycle in HeLa cells, possibly contributing to the development of new therapeutic strategies for cervix carcinoma.

IPP5, a novel inhibitor of protein phosphatase 1, suppresses tumor growth and progression of cervical carcinoma cells by inducing G2/M arrest.

Protein phosphatase 1 (PP1) is a major serine/threonine phosphatase that controls gene expression and cell cycle progression. Here, we report the characterization of a novel human bone marrow stromal cell (BMSC)-derived protein called protein phosphatase 1 inhibitor 5 (IPP5), which was obtained by large-scale random sequencing of a human BMSC cDNA library. The human IPP5 cDNA encodes a protein of 116 amino acid residues, which shares high homology with human protein phosphatase 1 inhibitor-1 (PPI-1). The effect of IPP5 on tumor growth and the underlying molecular mechanisms were investigated by overexpression of IPP5 in HeLa cells, a human cervical carcinoma cell line. Our results demonstrated that overexpression of the active mutant IPP5 inhibited the growth of HeLa cells both in vitro and in vivo. Biochemical analysis demonstrated that active mutant IPP5-mediated G2/M arrest of HeLa cells involved regulation of cyclin A1, cyclin B1, CDK1, p21, and p53, as well as increased inhibition of ERK activation. Furthermore, overexpression of the active mutant IPP5 leads to the formation of dikaryons following the failure of cytokinesis. Therefore, IPP5 might be a potential growth inhibitor for human tumor cells, especially for cervical carcinoma cells, and it could contribute to the development of new therapeutic strategies for human cervical cancer treatment.