The transcription factor CmERFI-2 represses CmMYB44 expression to increase sucrose levels in oriental melon fruit.

Soluble sugar accumulation in fruit ripening determines fleshy fruit quality. However, the molecular mechanism for this process is not yet understood. Here, we showed a transcriptional repressor, CmMYB44 regulates sucrose accumulation and ethylene synthesis in oriental melon (Cucumis. melo var. makuwa Makino) fruit. Overexpressing CmMYB44 suppressed sucrose accumulation and ethylene production. Furthermore, CmMYB44 repressed the transcriptional activation of CmSPS1 (sucrose phosphate synthase 1) and CmACO1 (ACC oxidase 1), two key genes in sucrose and ethylene accumulation, respectively. During the later stages of fruit ripening, the repressive effect of CmMYB44 on CmSPS1 and CmACO1 could be released by overexpressing CmERFI-2 (ethylene response factor I-2) and exogenous ethylene in "HS" fruit (high sucrose accumulation fruit). CmERFI-2 acted upstream of CmMYB44 as a repressor by directly binding the CmMYB44 promoter region, indirectly stimulating the expression level of CmSPS1 and CmACO1. Taken together, we provided a molecular regulatory pathway mediated by CmMYB44, which determines the degree of sucrose and ethylene accumulation in oriental melon fruit and sheds light on transcriptional responses triggered by ethylene sensing that enable the process of fruit ripening.

A urine-based liquid biopsy for detection of upper tract urothelial carcinoma: a self-matched study.

BACKGROUND: To establish the pathological diagnosis of UTUC before treatment is profitable. At present, the conventional pathological diagnostic methods have certain problems. Besides, the urine-based DNA methylation test have been already utilized to detect bladder cancer. OBJECTIVE: To evaluate the sensitivity and specificity of DNA methylation plus 17 genes mutation test and compare the combined test with cytology. MATERIALS AND METHODS: We included 45 patients from April 2019 to May 2022, all of whom underwent radical nephroureterectomy (RNU), nephrectomy, diagnostic ureteroscopy or tissue biopsy. Before surgery, the urine samples were collected for DNA methylation plus 17 genes mutation test and cytology. The test performance was calculated, and comparative ROC curves were drawn. RESULTS: The median age of the patients was 67 years. The Kappa value of the DNA methylation plus 17 genes mutation test and tissue pathology was 0.59 (p<0.001). The sensitivity/specificity/PPV/NPV of DNA methylation plus 17 genes mutation test was 86/80/94/62% compared with 29/100/100/29% for cytology. The AUC of DNA methylation plus 17 genes mutation test was 0.829 (p<0.001).The mutated gene proportion of UTUC patients was 51.43% for TERT and 25.71% for TP53. CONCLUSION: The test performance of DNA methylation plus 17 genes mutation test was satisfactory, which may replace cytology in the future. Further multicenter studies with larger samples are needed to confirm the clinical value of this promising method. NOVELTY & IMPACT STATEMENTS: We evaluated the diagnostic efficacy of a urine-based liquid biopsy for the detection of UTUC and compared the combined test with cytology. We found satisfactory results and concluded that the test could partly replace cytology. Further studies are needed.

Million Microfiber Releases: Comparing Washable and Disposable Face Masks.

The extensive use of single-use or disposable face masks has raised environmental concerns related to microfiber contamination. In contrast, research on the potential release and ecological impact of microfibers from washable masks (WMs), suggested as an eco-friendly alternative, is currently lacking. Here, we comprehensively investigated the release of microfibers from disposable and WMs of different types in simulated aquatic environments and real-life scenarios, including shaking, disinfection, hand washing, and machine washing. Using a combination of wide-field fluorescence microscopy, He-ion microscopy, and confocal µ-Raman spectroscopy, we revealed that disposable masks (DMs) released microfibers ranging from 18 to 3042 microfiber/piece, whereas WMs released 6.1 × 104-6.7 × 106 microfibers/piece depending on the simulated conditions above. Another noteworthy finding was the observed negative correlation between microfiber release and the proportion of reinforcement (embossing) on the DM surfaces. Microfibers from tested DMs primarily comprised polypropylene (PP), while WMs predominantly released poly(ethylene terephthalate) (PET) and cellulose microfibers. Furthermore, acute toxicological analyses unveiled that PP microfibers (0.01-50 mg/L) from DMs impacted zebrafish larval swimming behavior, while PET microfibers from WMs delayed early-stage zebrafish hatching. This study offers new insights into the source of microfiber contamination and raises concerns about the environmental implications linked to the use of washable face masks.

Rigorous Mapping of Data to Qualitative Properties of Parameter Values and Dynamics: A Case Study on a Two-Variable Lotka-Volterra System.

In this work, we describe mostly analytical work related to a novel approach to parameter identification for a two-variable Lotka-Volterra (LV) system. Specifically, this approach is qualitative, in that we aim not to determine precise values of model parameters but rather to establish relationships among these parameter values and properties of the trajectories that they generate, based on a small number of available data points. In this vein, we prove a variety of results about the existence, uniqueness, and signs of model parameters for which the trajectory of the system passes exactly through a set of three given data points, representing the smallest possible data set needed for identification of model parameter values. We find that in most situations such a data set determines these values uniquely; we also thoroughly investigate the alternative cases, which result in nonuniqueness or even nonexistence of model parameter values that fit the data. In addition to results about identifiability, our analysis provides information about the long-term dynamics of solutions of the LV system directly from the data without the necessity of estimating specific parameter values.

Segmental ureterectomy for high-risk ureteral carcinoma: a preliminary report.

BACKGROUND: EAU guidelines strongly recommend kidney sparing surgery (KSS) as the primary treatment option for the low-risk UTUC patients. While there are few reports involving the KSS treated for the high-risk counterparts, especially the ureteral resection. OBJECTIVE: To evaluate the effectiveness and safety of the segmental ureterectomy (SU) for the patients with high-risk ureteral carcinoma. MATERIALS AND METHODS: We included 20 patients from May 2017 to December 2021 who underwent segmental ureterectomy (SU) in Henan Provincial People's Hospital. The overall survival (OS) and progression free survival (PFS) were evaluated. Besides, the ECOG scores and postoperative complications were also included. RESULTS: As of December 2022, the mean OS was 62.1months (95%CI:55.6-68.6months) and the mean PFS was 45.0months (95%CI:35.9-54.1months). The median OS and median PFS were not reached. The 3-year OS rate was 70% and the 3-year PFS rate was 50%. The percentage of Clavien I and II complications was 15%. CONCLUSION: For the selected patients with high-risk ureteral carcinoma, the efficacy and safety of segmental ureterectomy were satisfactory. But we still need to conduct prospective or randomized study to validate the value of SU in patients with high-risk ureteral carcinoma.

Lightweight 3D Dense Autoencoder Network for Hyperspectral Remote Sensing Image Classification.

The lack of labeled training samples restricts the improvement of Hyperspectral Remote Sensing Image (HRSI) classification accuracy based on deep learning methods. In order to improve the HRSI classification accuracy when there are few training samples, a Lightweight 3D Dense Autoencoder Network (L3DDAN) is proposed. Structurally, the L3DDAN is designed as a stacked autoencoder which consists of an encoder and a decoder. The encoder is a hybrid combination of 3D convolutional operations and 3D dense block for extracting deep features from raw data. The decoder composed of 3D deconvolution operations is designed to reconstruct data. The L3DDAN is trained by unsupervised learning without labeled samples and supervised learning with a small number of labeled samples, successively. The network composed of the fine-tuned encoder and trained classifier is used for classification tasks. The extensive comparative experiments on three benchmark HRSI datasets demonstrate that the proposed framework with fewer trainable parameters can maintain superior performance to the other eight state-of-the-art algorithms when there are only a few training samples. The proposed L3DDAN can be applied to HRSI classification tasks, such as vegetation classification. Future work mainly focuses on training time reduction and applications on more real-world datasets.

mTORC2-AKT-LAT1 signalling participates in methionine-induced ß-CASEIN expression in mammary epithelial cells of dairy cows.

This study investigated the role of the mammalian target of rapamycin complex 2 (mTORC2)-protein kinase B (AKT) signalling in methionine (Met)-induced L-type amino acid transporter 1 (LAT1) expression and milk protein production. Primary mammary epithelial cells (MECs) from mammary parenchymal tissues of three lactating cows and MAC-T bovine MECs were cultured with or without 0.6 mM Met. Rapamycin-insensitive companion of mTOR (RICTOR) siRNA, the mTORC1 inhibitor rapamycin and the AKT activator SC79 were used to evaluate the effects of mTORC2-AKT signalling on Met-induced LAT1 expression and function. Each experiment was performed three times. Data were analysed with a two-sided unpaired t test or ANOVA with the Bonferroni multiple-comparison test. Western blotting showed that Met stimulation increased RICTOR expression (~244.67%; p < 0.05; control, 0.15 ± 0.026; Met, 0.517 ± 0.109) and AKT-S473 levels (~281.42%; p < 0.01; control, 0.253 ± 0.067; Met, 0.965 ± 0.019) in both primary MECs and MAC-T cells. Rapamycin-induced mTORC1 signalling inhibition decreased only Met-induced ß-CASEIN expression by ~21.24% (p < 0.01; Met, 0.777 ± 0.01; Met and rapamycin, 0.612 ± 0.04) and did not affect Met-stimulated AKT-S473 levels, suggesting that mTORC2-AKT activation upon Met stimulation also contributes to milk protein synthesis. LAT1 participates in Met-induced ß-CASEIN expression. In dairy cow MECs, mTORC2 inhibition by RICTOR siRNA decreased LAT1 levels on the plasma membrane by ~45.13% (p < 0.01; control, 0.359 ± 0.006; siRICTOR, 0.197 ± 0.004). However, SC79-induced AKT activation had the opposite effect (p < 0.01). In primary MECs and MAC-T cells, Met stimulation increased cytosolic and plasma membrane LAT1 expression respectively (MECs, 113.98% and 58.43%; MAC-T, 165.85% and 396.39%; p < 0.05). However, RICTOR siRNA significantly reduced Met-induced plasma membrane LAT1 expression (~76.48%; Met, 0.539 ± 0.05; Met and siRICTOR, 0.127 ± 0.012; p < 0.05). Thus, Met increased LAT1 expression and function via mTORC2-AKT signalling, upregulating milk protein synthesis in dairy cow MECs.

LncRNA PGM5-AS1 Inhibits the Progression of Bladder Cancer by Regulating miR-587/SLIT3 Axis.

Bladder cancer (BC), as one of the most common urological malignant tumor types worldwide, places a considerable burden on the economy and patients' health. Long non-coding RNA PGM5-AS1 has been shown to be downregulated in BC, however, its exact function in BC remains unclear. This study aimed to determine the influence of PGM5-AS1 on BC and its related mechanisms. The expression of PGM5-AS1, miR-587, and slit guided ligand 3 (SLIT3) in BC tissues and cells was detected using real-time quantitative polymerase chain reaction and Western blotting. In vitro functional experiments, including CCK-8, Transwell, and Western blotting, were used to assess BC cell proliferation, migration, and the expression of apoptosis-related proteins (Bax and Bcl-2). A xenograft tumor experiment was conducted to test the role of PGM5-AS1 in BC cell growth in vivo. In addition, the relationship between PGM5-AS1, miR-587, and SLIT3 was verified using luciferase reporter and RIP assays. PGM5-AS1 and SLIT3 were expressed at low levels in BC, whereas miR-587 exhibited the opposite trend. PGM5-AS1 overexpression significantly inhibited BC cell proliferation and migration, promoted apoptosis in vitro, and alleviated tumor growth in vivo. miR-587 has been shown to be a target of PGM5-AS1, and miR-587 overexpression can reverse the inhibitory effect of PGM5-AS1 upregulation on BC cell growth. Furthermore, miR-587 directly targeted SLIT3 and negatively regulated its expression. PGM5-AS1 inhibited BC cell proliferation and migration while facilitating apoptosis through the miR-587/SLIT3 pathway. PGM5-AS1 represses BC development via the miR-587/SLIT3 axis, indicating that PGM5-AS1 may be a candidate biomarker and target for BC treatment.

Transcription Factor CmNAC34 Regulated CmLCYB-Mediated ß-Carotene Accumulation during Oriental Melon Fruit Ripening.

Ripened oriental melon (Cucumis melo) with orange-colored flesh is rich in ß-carotene. Lycopene ß-cyclase (LCYB) is the synthetic enzyme that directly controls the massive accumulation of ß-carotene. However, the regulatory mechanism underlying the CmLCYB-mediated ß-carotene accumulation in oriental melon is fairly unknown. Here, we screened and identified a transcription factor, CmNAC34, by combining bioinformatics analysis and yeast one-hybrid screen with CmLCYB promoter. CmNAC34 was located in the nucleus and acted as a transcriptional activator. The expression profile of CmNAC34 was consistent with that of CmLCYB during the fruit ripening. Additionally, the transient overexpression of CmNAC34 in oriental melon fruit promoted the expression of CmLCYB and enhanced ß-carotene concentration, while transient silence of CmNAC34 in fruit was an opposite trend, which indicated CmNAC34 could modulate CmLCYB-mediated ß-carotene biosynthesis in oriental melon. Finally, the yeast one-hybrid (Y1H), electrophoretic mobility shift assay (EMSA), ß-glucuronidase (GUS) analysis assay, and luciferase reporter (LUC) assay indicated that CmNAC34 could bind to the promoter of CmLCYB and positively regulated the CmLCYB transcription level. These findings suggested that CmNAC34 acted as an activator to regulate ß-carotene accumulation by directly binding the promoter of CmLCYB, which provides new insight into the regulatory mechanism of carotenoid metabolism during the development and ripening of oriental melon.

Cell death-inducing DNA fragmentation factor-α-like effector C (CIDEC) regulates acetate- and ß-hydroxybutyrate-induced milk fat synthesis by increasing FASN expression in mammary epithelial cells of dairy cows.

Increasing acetate and ß-hydroxybutyrate (BHB) supply to lactating cows will increase milk fat synthesis. However, the underlying molecular mechanism remains largely unknown. Cell death-inducing DNA fragmentation factor-α-like effector C (CIDEC) is a lipid droplet-associated protein that promotes intracellular triacylglycerol accumulation. In the present study, using gene overexpression and knockdown, we detected the contributions of CIDEC on milk fat synthesis in mammary epithelial cells of dairy cows in the presence of acetate and BHB. The results showed that knockdown of CIDEC decreased fatty acid synthase (FASN) expression and intracellular triacylglycerol content, whereas overexpression of CIDEC had the opposite effect. The transcription factor CCAAT/enhancer-binding protein ß (C/EBPß) regulates cell growth and differentiation in the mammary gland. We demonstrated that the FASN promoter had a canonical C/EBPß binding sequence. CEBPB overexpression upregulated FASN expression and milk fat synthesis, whereas CEBPB knockdown had the opposite effect. Moreover, knockdown of CEBPB attenuated the promoting effects of CIDEC on acetate- and BHB-induced FASN transcription. Taken together, our data showed that acetate and BHB induced FASN expression in mammary epithelial cells of dairy cows in a CIDEC-C/EBPß-dependent manner, which provides new insights into the understanding of the molecular events involved in milk fat synthesis.

ERK1/2-PPARγ pathway is involved in Chlamydia pneumonia-induced human umbilical vein endothelial cell apoptosis through increased LOX-1 expression.

Chlamydia pneumonia (C.pn) is a common respiratory pathogen that is involved in human cardiovascular diseases and promotes the development of atherosclerosis in hyperlipidemic animal models. C.pn reportedly up-regulated lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in endothelial cells. Recently, the anti-atherosclerotic activity of peroxisome proliferator-activated receptor γ (PPARγ) has been documented. In the present study, we investigated the effect of C.pn on LOX-1 expression in human umbilical vein endothelial cells (HUVECs) and identified the involvement of the PPARγ signaling pathway therein. The results showed that C.pn increased the expression of LOX-1 in HUVECs in a dose- and time-dependent manner. C.pn-induced up-regulation of LOX-1 was mediated by ERK1/2, whereas p38 MAPK and JNK had no effect on this process. C.pn induced apoptosis, inhibited cell proliferation, and decreased the expression PPARγ in HUVECs. Additionally, LOX-1 activity and cell injury caused by C.pn through activation of ERK1/2 was completely inhibited by rosiglitazone, a PPARγ agonist. In conclusion, we inferred that activation of PPARγ in HUVECs suppressed C.pn-induced LOX-1 expression and cell damage by inhibiting ERK1/2 signaling.

Curcumin suppresses the proliferation and tumorigenicity of Cal27 by modulating cancer-associated fibroblasts of TSCC.

Cancer-associated fibroblasts (CAFs) are "activated" fibroblasts in the tumor microenvironment (TME) and play a vital role in all steps of cancer development. Increasing evidence focusing on the function of CAFs suggests that CAFs are candidate therapeutic targets and that drugs targeting the modification of CAFs would suppress tumor progression and be beneficial to tumor treatment and prevention. In the present study, we found that curcumin reversed the phenotype of CAFs to that of peri-tumor fibroblast (PTF)-like cells by downregulating the expression of α-SMA (a special marker for CAFs) and inhibiting the secretion of pro-carcinogenic cytokines, including transforming growth factor-ß1 (TGF-ß1), matrix metalloproteinases 2 (MMP2), and stromal cell-derived factor-1 (SDF-1). We further demonstrated that the conditioned medium (CM) derived from CAFs promoted the proliferation of Cal27, and this effect was confirmed by the xenograft model. More importantly, we found that curcumin blocked the CAF-mediated enhancement of Cal27 proliferation in vitro and in vivo. In conclusion, our data suggest that curcumin reverses cell phenotype from CAF to PTF-like cells and suppresses the CAF-mediated proliferation and tumorigenicity of Cal27 by inhibiting TSCC CAFs.

Unbalanced expression of membrane-bound and soluble inducible costimulator and programmed cell death 1 in patients with myasthenia gravis.

This study aimed to investigate the possible functions and mechanisms of positive and negative costimulatory molecules in the pathological process of myasthenia gravis (MG). The expression levels of membrane-bound inducible costimulator (ICOS) and programmed cell death 1 (PD-1) in peripheral blood T cells, their corresponding ligands ICOSL and PDL-1 on B cells, and their soluble forms (sICOS, sPD-1, sICOSL, and sPDL-1) in plasma were detected in patients with untreated-stage MG (USMG) and remission-stage MG (RSMG). The results showed that the expression levels of membrane-bound ICOS and PD-1 in the peripheral blood T cells of the USMG group and their corresponding ligands ICOSL and PD-L1 on B cells were significantly increased compared to those in the RSMG group and healthy controls (HCs). The levels of sICOSL and sPD-1 were significantly upregulated in USMG patients compared to those in the RSMG and HC groups, while the levels of sICOS and sPD-L1 were not different. The expression of PD-L1 on CD19+ B cells was positively correlated with the concentrations of AchR Ab in the USMG group. The expression of ICOS and PD-1 in CD4+ T cells and the expression of ICOSL and PD-L1 on CD19+ B cells were positively correlated with the quantitative myasthenia gravis (QMG) scores in the USMG group. Also, in the USMG group, the plasma levels of sICOSL and sPD-1 were positively correlated with the QMG scores. In addition, the percentage of peripheral blood follicular helper T (Tfh) cells in the USMG group was positively correlated with ICOS and PD-1 expression on CD4+ T cells and ICOSL and PD-L1 expression on CD19+ B cells. There were positive correlations between sICOSL and sPD-1 levels and the percentage of peripheral blood Tfh cells and plasma interleukin-21 (IL-21) levels in the USMG group. The results suggest that the positive ICOS/ICOSL and negative PD-1/PD-L1 costimulatory molecule pairs participate in the pathological process of MG. Abnormal sICOSL and sPD-1 expression might interfere with the normal signal transduction of ICOS and PD-1 on Tfh cells, causing excessive activation of Tfh cells and promotion of disease progression. sICOSL and sPD-1 have potential value in monitoring MG disease states.

Association of exposure to Chinese famine in early life with the incidence of hypertension in adulthood: A 22-year cohort study.

BACKGROUND AND AIMS: Undernutrition in early life may have a lifelong effect on adult health. The conclusions on the association of exposure to famine with the risk of hypertension were inconsistent. The aim of this study was to examine the association of exposure to the Chinese famine with incident hypertension. METHODS AND RESULTS: Data were obtained from the China Health and Nutrition Survey. All included participants were divided into five birth cohorts: no exposure, born in or after 1962 (N = 2 088); fetal exposure, between 1959 and 1961 (N = 880); early childhood exposure, between 1956 and 1958 (N = 1 214); mid-childhood exposure, between 1953 and 1955 (N = 1 287); and late childhood exposure, between 1949 and 1952 (N = 1 445). Hypertension was defined as SBP/DBP ≥140/90 mmHg, use of hypertensive medications, or a self-reported diagnosis. A total of 6 914 participants were included. The exposure to famine decreased the incidence of hypertension (P = 0.0018, 0.0001, <0.0001, and <0.0001; HR: 0.715, 0.686, 0.622, and 0.527, respectively) in males. Similarly, the exposure to famine might also decrease incident hypertension in the rural areas (P = 0.0013, <0.0001, <0.0001, and <0.0001; HR: 0.735, 0.706, 0.679, and 0.539, respectively). There were interaction effects between famine severity and exposure to famine in early (P = 0.024) and late childhood (P = 0.009). CONCLUSION: Exposure to the Chinese famine decreased the incidence of hypertension, especially in males and in the rural areas. Furthermore, the exposure postponed the age at the onset of hypertension.

Qiliqiangxin Protects against Renal Injury in Rat with Cardiorenal Syndrome Type I through Regulating the Inflammatory and Oxidative Stress Signaling.

Cardiorenal syndrome (CRS) is a frequently encountered clinical condition when the dysfunction of either the heart or kidneys amplifies the failure progression of the other organ. CRS remains a major global health problem. Qiliqiangxin (QLQX) is a traditional Chinese herbs medication, which can improve cardiac function, urine volume, and subjective symptoms in patients with chronic heart failure. In the present study, we aim to investigate the role of QLQX in the treatment of CRS type I and the possible mechanism through establishment of a rat model of myocardial infarction. Rats in CRS-Q group were orally treated with QLQX daily for 2 weeks or 4 weeks, while in sham group and CRS-C group were treated with saline at the same time. Enzyme-linked immunosorbent assay (ELISA) analysis showed that QLQX significantly reduced the levels of angiotensin II (AngII), brain natriuretic peptides (BNP), creatinine (CRE), cystatin C (CysC), tumor necrosis factor (TNF)-α, interleukin (IL)-6, microalbuminuria (MAU), and neutrophil gelatinase-associated lipocalin (NGAL) in plasma induced by myocardial infarction. Western blot analysis showed that QLQX significantly reduced the expressions of AngII, non-phagocytic cell oxidase (NOX)2, and B-cell lymphoma (Bcl)2 associated X protein (Bax), and increased the expressions of Bcl2 and Angiotensin II Type 1 receptor (ATR) in the kidney as compared with the CRS-C group. Fluorescence microscopy showed that the content of reactive oxygen species (ROS) was significantly reduced in the kidney as compared with the CRS-C group. We also examined the apoptosis level in kidney by using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining, and the result showed that QLQX significantly reduced the apoptosis level in kidney induced by myocardial infarction. Taken together, we suggest that QLQX may be a potentially effective drug for the treatment of CRS by regulating inflammatory/oxidative stress signaling.

The effects of L-type amino acid transporter 1 on milk protein synthesis in mammary glands of dairy cows.

The mammary gland requires the uptake of AA for milk protein synthesis during lactation. The L-type amino acid transporter 1 (LAT1, encoded by SLC7A5), found in many different types of mammalian cells, is indispensable as a transporter of essential AA to maintain cell growth and protein synthesis. However, the function of LAT1 in regulating milk protein synthesis in the mammary gland of the dairy cow remains largely unknown. For the current study, we characterized the relationship between LAT1 expression and milk protein synthesis in lactating dairy cows and investigated whether the mammalian target of rapamycin complex 1 (mTORC1) signaling controls the expression of LAT1 in their mammary glands. We found that LAT1 and the heavy chain of its chaperone, 4F2, were expressed in mammary tissues of lactating cows, with the expression levels of LAT1 and the 4F2 heavy chain being significantly greater in lactating mammary tissues with high-milk protein content (milk yield, 33.8 ± 2.1 kg/d; milk protein concentration >3%, wt/vol,; n = 3) than in tissues from cows with low-milk protein content (milk yield, 33.7 ± 0.5 kg/d; milk protein concentration <3%, wt/vol; n = 3). Immunofluorescence staining of sectioned mammary tissues from cows with high and low milk protein content showed that LAT1 was located on the whole plasma membrane of alveolar epithelial cells, suggesting that LAT1 provides essential AA to the mammary gland. In cultured mammary epithelial cells from the dairy cows with high-milk protein content, knockdown of LAT1 expression decreased cell viability and ß-casein expression; in contrast, overexpression of LAT1 had the opposite effect. Inhibition of mTORC1 by rapamycin attenuated the phosphorylation of molecules related to mTORC1 signaling and caused a marked decrease in LAT1 expression in the cultured cells; expression of ß-casein also decreased significantly. These results suggest that LAT1 is involved in milk protein synthesis in the mammary glands of lactating dairy cows and that the mTORC1 signaling pathway might be a control point for regulation of LAT1 expression, which could ultimately be used to alter milk protein synthesis.

PA-MSHA induces apoptosis and suppresses metastasis by tumor associated macrophages in bladder cancer cells.

BACKGROUND: The aim of the present study was to investigate effects of Pseudomonas aeruginosa-mannose-sensitive hemagglutinin (PA-MSHA) on the inhibition of the proliferation of bladder cancer cell lines and to further define its functional mechanisms. METHODS: A rat model of bladder tumor was induced by intravesical N-methyl-N nitrosourea. The dynamic growth of tumor was measured by whole-body fluorescent imaging system. Morphological analysis was observed by hematoxylin-eosin staining and microscopic examination. The expression of Caspase 3 and E-Ca were detected by immunohistochemistry technique. Macrophages were separated by flow cytometry. The expression of cytokines was measured by qRT-PCR and western blot. Apoptosis ability was conducted by means of annexin V and propidium iodide. The abilities of invasion and migration were determined by transwell migration assay and scratch assay. RESULTS: PA-MSHA and PA-MSHA + Fisetin groups inhibited the growth of tumor and increased the ratio of M1/M2. For one thing, PA-MSHA suppressed the invasive ability of the bladder tumor cell and promoted bladder tumor cell apoptosis. For another, it facilitated the expression of M1 cytokines and reduced expression of M2 cytokines. Furthermore, treated with PA-MSHA, mouse M1 phagocytosis rates were higher than that of M2 macrophages for bladder cancer lines. CONCLUSIONS: The data revealed that PA-MSHA might promote apoptosis and inhibit proliferation, invasion and migration of mouse bladder cancer cells by inducing M1 polarization.

The effects of cell death-inducing DNA fragmentation factor-α-like effector C (CIDEC) on milk lipid synthesis in mammary glands of dairy cows.

Adequate lipid synthesis by the mammary gland during lactation is essential for the survival of mammalian offspring. Cell death-inducing DNA fragmentation factor-α-like effector C (CIDEC) is a lipid droplet-associated protein and functions to promote lipid accumulation and inhibit lipolysis in mice and human adipocytes. However, the function of CIDEC in regulation of milk lipid synthesis in dairy cow mammary gland remains largely unknown. In this study, 6 multiparous Holstein cows (parity = 3) in early lactation were allocated to high-fat milk (milk yield 33.9 ± 2.1 kg/d, milk fat >3.5%, n = 3) and low-fat milk (milk yield 33.7 ± 0.5 kg/d, milk fat <3.5%, n = 3) groups according to their milk fat content. Lactating cows were slaughtered at 90 d in milk and mammary tissues were collected to detect CIDEC localization. Immunofluorescence staining of sections of lactating mammary glands with high- and low-fat milk showed that CIDEC was expressed in the cytoplasm of epithelial cells and localized to lipid droplets. Lipid droplets and CIDEC protein were also detected in isolated lactating mammary epithelial cells of dairy cows. Immunostaining of CIDEC in isolated mammary epithelial cells also confirmed its presence in the nucleus. The knockdown of CIDEC in cultured bovine mammary epithelial cells decreased milk lipid content and reduced expression of genes associated with mammary de novo fatty acid synthesis, short- and long-chain intracellular fatty acid activation, triacylglycerol synthesis, and transcription regulation. These genes included those for acetyl-CoA carboxylase (ACC, -60%), fatty acid synthase (FASN, -65%), acyl-CoA synthetase short-chain family member 2 (ACSS2, -50%), acyl-CoA synthetase long-chain family member 1 (ACSL1, -30%), diacylglycerol acyltransferase 1 (DGAT1, -60%), sterol regulatory element-binding protein 1 (SREBP1, -45%), and SREBP cleavage activating protein (SCAP, -66%). Conversely, in cells overexpressing CIDEC, triacylglycerol content was increased, and transcription of those genes involved in milk lipid synthesis was coordinately upregulated. These results suggest that CIDEC plays an important role in regulating milk lipid synthesis in dairy cow mammary gland via a mechanism involving gene expression, which provides further insight into the mechanisms regulating mammary lipogenesis in ruminants.

Spleen tyrosine kinase regulates mammary epithelial cell proliferation in mammary glands of dairy cows.

Spleen tyrosine kinase (SYK) is a nonreceptor tyrosine kinase that has been considered a hematopoietic cell-specific signal transducer involved in cell proliferation and differentiation. However, the role of SYK in normal mammary gland is still poorly understood. Here we show that SYK is expressed in mammary glands of dairy cows. Expression of SYK was higher in dry period mammary tissues than in lactating mammary tissues. Knockdown and overexpression of SYK affected dairy cow mammary epithelial cell proliferation as well as the expression of signal molecules involved in proliferation, including protein kinase B (PKB, also known as AKT1), p42/44 mitogen-activated protein kinase (MAPK), and signal transducer and activator of transcription 5 (STAT5). Dual-luciferase reporter assay showed that SYK increased the transcriptional activity of the AKT1 promoter, and cis-elements within the AKT1 promoter region from -439 to -84 bp mediated this regulation. These results suggest that SYK affects mammary epithelial cell proliferation by activating AKT1 at the transcriptional level in mammary glands of dairy cows, which is important for the mammary remodeling process in dry cows as well as for increasing persistency of lactation in lactating cows.

Hydrolyzable tannins and pectin are glycolipid lowering ingredients in Phyllanthus emblica Linn.: Valid interaction with gut action targets.

Phyllanthus emblica L., a distinctive fruit, is rich in polyphenols and polysaccharide. However, there is a lack of knowledge regarding the role of these compounds as glycolipid lowering ingredients. In this study, the glycolipid lowering ingredients and their effects have been determined by gradually comparing varieties, parts, and components, splitting components, and calculating combined index via their interactions with digestion enzymes, bile acids, cholesterol micelles and probiotics. Results indicated that the glycolipid lowering ingredients were polyphenols and polysaccharide, which located in the pulp, and not influenced by the variety. Pectin with multiple structural domains, and hydrolyzable tannins, i.e. gallic acid, ellagic acid and their derivatives interacted with multiple gut action targets could regulate glycolipid digestion, absorption and metabolism. Polysaccharide and polyphenols demonstrated a synergistic effect in lowering glycolipid by interacting with gut action targets. These findings highlight the potential for further investigation and utilization of glycolipid lowering ingredients in fruits.