Ulinastatin inhibits the metastasis of nasopharyngeal carcinoma by involving uPA/uPAR signaling.

Distant metastasis is the primary reason for treatment failure in patients with nasopharyngeal carcinoma (NPC). In this study, we investigated the effect of ulinastatin (UTI) on NPC metastasis and its underlying mechanism. Highly-metastatic NPC cell lines S18 and 58F were treated with UTI and the effect on cell proliferation, migration, and invasion were determined by MTS and Transwell assays. S18 cells with luciferase-expressing (S18-1C3) were injected into the left hind footpad of nude mice to establish a model of spontaneous metastasis from the footpad to popliteal lymph node (LN). The luciferase messenger RNA (mRNA) was measured by quantitative polymerase chain reaction (qPCR), and the metastasis inhibition rate was calculated. Key molecular members of the UTI-related uPA, uPAR, and JAT/STAT3 signaling pathways were detected by qPCR and immunoblotting. UTI suppressed the migration and infiltration of S18 and 5-8F cells and suppressed the metastasis of S18 cells in vivo without affecting cell proliferation. uPAR expression decreased from 24 to 48 h after UTI treatment. The antimetastatic effect of UTI is partly due to the suppression of uPA and uPAR. UTI partially suppresses NPC metastasis by downregulating the expression of uPA and uPAR.

Changes in cardiovascular function and related psychophysiological pathways in a rat model of post-traumatic stress disorder.

This study was aimed to investigate the cardiovascular function in rats with post-traumatic stress disorder (PTSD) and the potential association with the activities of the rostral ventrolateral medulla (RVLM) and the medial habenular nucleus (MHb). Multi-channel in vivo recordings were used to simultaneously acquire spontaneous neuronal firing and peripheral physiological indices, and FluoroGold (FG) retrograde tracing technique was used to observe the projections of labeled neurons in the MHb. The results showed that the discharge frequency of RVLM and MHb neurons, the systolic blood pressure (SBP), and the mean arterial pressure (MAP) in the PTSD group were all increased significantly compared with those in control group (P < 0.05). MHb neurons were retrogradely labeled by FG through microinjection (4% FG, 0.5 µL) into the RVLM. In the control group, electrical stimulation in the MHb increased heart rate (HR) at 100-300 µA (P < 0.05), elevated SBP and MAP at 200-300 µA (P < 0.05), and remarkably increased the RVLM neuronal discharge frequency at 100-500 µA (P < 0.05 or P < 0.01). In the PTSD group, however, only the discharge frequency of RVLM neurons was increased by the electrical stimulation at 100-300 µA (P < 0.05). These results suggest that cardiovascular activities of the PTSD model rat are enhanced, and this change may be related to the activity changes of RVLM and MHb and the potential connection between the two nuclei.

Synthesis, structure and magnetocaloric properties of a new two-dimensional gadolinium(III) coordination polymer based on azobenzene-2,2',3,3'-tetracarboxylic acid.

A new Gd3+ coordination polymer (CP), namely, poly[diaqua[µ4-1'-carboxy-3,3'-(diazene-1,2-diyl)dibenzene-1,2,2'-tricarboxylato]gadolinium(III)], [Gd(C16H7N2O8)(H2O)2]n, (I), has been synthesized hydrothermally from Gd(NO3)3·6H2O and azobenzene-2,2',3,3'-tetracarboxylic acid (H4abtc). The target solid has been characterized by single-crystal and powder X-ray diffraction, elemental analysis, IR spectroscopy and susceptibility measurements. CP (I) crystallizes in the monoclinic space group C2/c. The structure features a 4-connected topology in which Gd3+ ions are connected by carboxylate groups into a linear chain along the monoclinic symmetry direction. Adjacent one-dimensional aggregates are bridged by Habtc3- ligands to form a two-dimensional CP in the (10-1) plane. A very short hydrogen bond [O...O = 2.4393 (4) Å] links neighbouring layers into a three-dimensional network. A magnetic study revealed antiferromagnetic Gd...Gd coupling within the chain direction. CP (I) displays a significant magnetocaloric effect (MCE), with a maximum -ΔSm of 27.26 J kg-1 K-1 for ΔH = 7 T at 3.0 K. As the MCE in (I) exceeds that of the commercial magnetic refrigerant GGG (Gd3Ga5O12, -ΔSm = 24 J kg-1 K-1, ΔH = 30 kG), CP (I) can be regarded as a potential cryogenic material for low-temperature magnetic refrigeration.

How to Identify Patients at High Risk of Developing Nasal-Type, Extranodal Nature Killer/T-Cell Lymphoma-Associated Hemophagocytic Syndrome.

Nasal-type, extranodal nature killer (NK)/T-cell lymphoma-associated hemophagocytic syndrome (NK/T-LAHS) is a rare and life-threatening disease, requiring investigation of risk stratification. We conducted a retrospective study and proposed nomograms to predict NK/T-LAHS. The discriminative ability and calibration of the nomograms for prediction were tested using C statistics and calibration plots. We analyzed 533 patients with extranodal NK/T-cell lymphoma (ENKTL), out of which 71 were diagnosed with hemophagocytic syndrome (HPS), with a cumulative incidence of 13.3%. Significant difference for 2-year survival was found between patients with and without HPS (14.7% vs. 77.5%). Analyses showed that Eastern Cooperative Oncology Group (ECOG) performance status (PS) ≥2, B symptoms, and bone marrow (BM) invasion were significantly associated with NK/T-LAHS. We used these data as the basis to establish a nomogram of risk index for ENKTL (RINK). In 335 patients with available data for Epstein-Barr virus DNA (EBV-DNA), we found high viral copies (≥4,450 copies/ml) were correlated with NK/T-LAHS. When these data were added to RINK, we developed another nomogram that included EBV-DNA data (RINK-E). The nomograms displayed good accuracy in predicting NK/T-LAHS with a C-statistics of 0.919 for RINK and a C-statistics of 0.946 for RINK-E, respectively. The calibration chart also showed an excellent consistency between the predicted and observed probabilities. The proposed nomograms provided individualized risk estimate of HPS in patients with ENKTL.

Dihydrochalcones from the leaves of Lithocarpus litseifolius.

Three new phlorizin derivatives, 6"-O-vanilloylphlorizin (1), 6"-O-(4-hydroxybenzoyl)phlorizin (2), 6"-O-feruloylphlorizin (3), along with four known dihydrochalcones, phlorizin (4), 3-hydroxyphlorizin, trilobatin, and 6"-O-acetylphlorizin were isolated from the leaves of Lithocarpus litseifolius. Their structures were established by analysis of extensive spectroscopic data. The new compounds were shown to be non-cytotoxic when tested against A549, HeLa, HepG2, and MCF-7 cell lines.

Fe3O4-assisted laser desorption ionization mass spectrometry for typical metabolite analysis and localization: Influencing factors, mechanisms, and environmental applications.

Fe3O4 has been suggested as an efficient matrix for small-molecule analysis by laser desorption ionization mass spectrometry (LDI-MS), but thus far there has been no systematic study exploring the influencing factors of nano-Fe3O4 on the detection of typical metabolites, or the mechanism by which nano-Fe3O4 assists the desorption and ionization of analytes after receiving laser energy. In this study, Fe3O4 nanoparticles with different physicochemical properties were synthesized and characterized. The results revealed that smaller particle size and greater surface hydroxyl amount of nano-spherical Fe3O4 could improve the intensity and relative standard deviation of typical metabolites by LDI-MS. The thermally driven desorption process played a vital role in LDI performance, but the chemical interactions between nano-Fe3O4 and analytes did not. Good intra- or inter-spot repeatability and linearity of analytes were obtained by the optimum Fe3O4-assisted LDI-MS. Finally, the developed method was successfully used for the rapid analysis and localization of endogenous metabolites in biofluids and whole zebrafish tissue section samples. Our results not only elucidate the influencing factors and mechanisms of nano-Fe3O4 for the detection of typical metabolites in LDI-MS but also reveal an innovative tool for the imaging of chemicals in the regions of interest in terms of eco-toxicological research.

[Effects of different nitrogen fertilizers on annual emissions of greenhouse gas from maize field in Northeast China]. / 不同氮肥对东北春玉米农田温室气体周年排放的影响.

A field experiment was conducted to investigate the greenhouse gas emission at high latitude farmland in Northeast China. We monitored the greenhouse gas emission using the static chamber-gas chromatography method. Four nitrogen (N) fertilizers were used: conventional N fertilization (CN), slow release fertilizer (SLN), urea plus nitrification inhibitor and urease inhibitor (NIUI), and no nitrogen fertilizer (NN). The results showed that the yields under CN, SLN and NIUI treatments were 9618, 9376 and 9645 kg·hm-2, respectively. Compared with CN treatment, SLN increased soil N2O emission in the growing season and decreased N2O emission in the non-growing season. The cumulative N2O emission flux of NIUI treatment was 39.0% lower than that of CN. There were no significant differences in the annual cumulative CO2 emission flux among the treatments. The spring maize field in Northeast China was a weak CH4 sink. NIUI treatment promoted soil CH4 absorption in maize growing season compared with CN treatment. In summary, urea plus nitrification inhibitor and urease inhibitor could significantly reduce soil greenhouse gas emissions with high yield of maize.

Analysis of transcriptional response in zebrafish eleutheroembryos exposed to climbazole: Signaling pathways and potential biomarkers.

Climbazole is an antifungal active ingredient used in personal care products. After application this chemical reaches the aquatic environment and may pose a risk to fish. In the present study, we measured the transcriptional effects of essential genes related to a wide range of signaling pathways on zebrafish eleutheroembryos exposed to climbazole at environmentally relevant and predicted worst-case environmental concentrations, and explored the potential biomarkers via partial least squares discriminant analysis. Transcription analysis covering up to 73 genes revealed significant down-regulation of circadian rhythm- and steroidogenesis-related genes in zebrafish embryos and larvae after exposure to environmentally relevant concentrations of climbazole. This topical antifungal agent also modulated the transcripts of genes involved in inflammation, oxidative stress, oocyte maturation, and sexual differentiation at predicted worst-case environmental concentrations. In addition, mprα, igf3, nr1d1, nr1d2b, cyp19a1a, vtg1, il-1ß, and il-8 were chosen as potential biomarkers in embryonic zebrafish following exposure to climbazole. These findings can help us understand the remarkable transcriptional response to climbazole in the early life stage of zebrafish. Future research should elucidate whether the transcriptional modulation translates into metabolic phenotypes associated with the corresponding signaling pathways. Environ Toxicol Chem 2019;38:794-805. © 2019 SETAC.

Global expression profiling and pathway analysis of mouse mammary tumor reveals strain and stage specific dysregulated pathways in breast cancer progression.

It is believed that the alteration of tissue microenvironment would affect cancer initiation and progression. However, little is known in terms of the underlying molecular mechanisms that would affect the initiation and progression of breast cancer. In the present study, we use two murine mammary tumor models with different speeds of tumor initiation and progression for whole genome expression profiling to reveal the involved genes and signaling pathways. The pathways regulating PI3K-Akt signaling and Ras signaling were activated in Fvb mice and promoted tumor progression. Contrastingly, the pathways regulating apoptosis and cellular senescence were activated in Fvb.B6 mice and suppressed tumor progression. We identified distinct patterns of oncogenic pathways activation at different stages of breast cancer, and uncovered five oncogenic pathways that were activated in both human and mouse breast cancers. The genes and pathways discovered in our study would be useful information for other researchers and drug development.

Along with its favorable prognostic role, CLCA2 inhibits growth and metastasis of nasopharyngeal carcinoma cells via inhibition of FAK/ERK signaling.

BACKGROUND: CLCA2 was reported as a tumor suppressor and disregulated in breast cancer. However, its function in tumor growth and metastasis in NPC has rarely been reported. In this study, we investigated the functional and molecular mechanisms by which CLCA2 influences NPC. METHODS: CLCA2 expression in human NPC cell lines and tissues was examined via real-time PCR (RT-PCR), Western blot and IHC. The biological roles of CLCA2 in proliferative, migration and invasion of NPC cell lines was evaluated in 5-8F, S18, S26 and SUNE-1 cells. Cell viability, migration and invasion were assessed in vitro by MTS, colony formation and transwell assay, respectively. CLCA2 in growth and metastasis of NPC were evaluated in vivo through NPC xenograft tumor growth, lung metastatic mice model and popliteal lymph node (LN) metastasis model. RESULTS: Overexpression of CLCA2 significantly decreased proliferation, migration and invasion of NPC cells. In contrast, knockdown of CLCA2 elicited the opposite effects. CLCA2 overexpression suppressed xenograft tumor growth and lung, popliteal lymph node (LN) metastasis in vivo. CLCA2 inhibited tumor metastasis through suppressing epithelial-Mesenchymal transition (EMT) and in-activating FAK/ERK1/2 signaling pathway in NPC cells. Immunohistochemical staining of 143 NPC samples revealed that CLCA2 expression was an independent, favorable prognostic factor for overall survival and distant metastasis-free survival of patients. In addition, inhibition of FAK and ERK1/2 reversed CLCA2 silencing-induced tumor cell migration. Furthermore, inhibitors against chloride channels suppressed NPC cellular migration which could have been enhanced by the presence of CLCA2. CONCLUSION: CLCA2 suppress NPC proliferation, migration, invasion and epithelial-mesenchymal transition through inhibiting FAK/ERK signaling.

Association of choroidal thickness with early stages of diabetic retinopathy in type 2 diabetes.

AIM: To assess the correlation between choroidal thickness (CT) and the early stages of diabetic retinopathy (DR) in type 2 diabetic patients. METHODS: We divided 83 diabetic patients (51-80 years of age; 50 females) into non diabetic retinopathy group (NDR) and mild/moderate nonproliferative diabetic retinopathy (NPDR) group, and compared them with 26 non-diabetic control subjects (51-78 years of age; 16 females). Subfoveal choroidal thickness (SFCT) and parafoveal choroidal thickness (PFCT) were measured using enhanced depth imaging spectral-domain optical coherence tomography (EDI-OCT). Ocular health status, disease duration, body mass index, and hemoglobin A1c (HbA1c) were recorded. RESULTS: The mean ages of the NDR, NPDR, and control groups were 68.0±6.9y, 67.8±6.4y, and 65.1±6.3y, respectively (P=0.17). Pearson correlation of the right and left eyes for the control subjects was 0.95 and for the NDR subjects was 0.93. SFCT for the right eyes of the controls was 252.77± 41.10 µm, which was significantly thicker than that of the right eyes in NDR group (221.51±46.56 µm) and the worse eyes of the NPDR group (207.18±61.87 µm; ANOVA, P<0.01). In the diabetic patients pooled together, age was the only variable significantly associated with SFCT (multiple linear regression analysis, P=0.01). CONCLUSION: CT decreased significantly in the NDR and mild/moderate NPDR eyes compared with the control eyes. Age is significantly associated with SFCT in the diabetic patients. Diabetic choroidopathy may be present before clinical retinopathy.

Postharvest temperature influences volatile lactone production via regulation of acyl-CoA oxidases in peach fruit.

The biosynthesis of volatile compounds in plants is affected by environmental conditions. Lactones are considered to be peach-like aroma volatiles; however, no enzymes or genes associated with their biosynthesis have been characterized. White-fleshed (cv. Hujingmilu) and yellow-fleshed (cv. Jinxiu) melting peach (Prunus persica L. Batsch) fruit were used as materials in two successive seasons and responses measured to four different temperature treatments. Five major lactones accumulated during postharvest peach fruit ripening at 20 °C. Peach fruit at 5 °C, which induces chilling injury (CI), had the lowest lactone content during subsequent shelf life after removal, while 0 °C and a low-temperature conditioning (LTC) treatment alleviated development of CI and maintained significantly higher lactone contents. Expression of PpACX1 and activity of acyl-CoA oxidase (ACX) with C16-CoA tended to increase during postharvest ripening both at 20 °C and during shelf life after removal from cold storage when no CI was developed. There was a positive correlation between ACX and lactones in peach fruit postharvest. Changes in lactone production in response to temperatures are suggested to be a consequence of altered expression of PpACX1 and long-chain ACX activity.

Expression of genes associated with aroma formation derived from the fatty acid pathway during peach fruit ripening.

Changes in characteristic aroma volatiles, levels of fatty acids as aroma precursors, and expression patterns of related genes, including lipoxygenase (LOX), hydroperoxide lyase (HPL), alcohol dehydrogenase (ADH), alcohol acyltransferase (AAT), and fatty acid desaturase (FAD), were studied in peach ( Prunus persica L. Batsch., cv. Yulu) fruit during postharvest ripening at 20 degrees C. Concentrations of n-hexanal, (E)-2-hexenal, (E)-2-hexenol, and (Z)-3-hexenol decreased, whereas the production of (Z)-3-hexenyl acetate, gamma-hexalactone, gamma-octalactone, gamma-decalactone, and delta-decalactone increased with fruit ripening. Lactones showed a clear pattern concomitant with the climacteric rise in ethylene production, with gamma-decalactone being the principal volatile compound at the late ripening stage. Of the LOX family genes, PpLOX2 and PpLOX3 had relatively high transcript levels initially followed by a decline with fruit ripening, while levels of PpLOX1 and PpLOX4 transcripts were upregulated by accumulated ethylene production. Expression of PpHPL1, PpADH1, PpADH2, and PpADH3 showed similar decreasing patterns during ripening. Expression levels of PpAAT1 showed a rapid increase during the first 2 days of postharvest ripening followed by a gradual decrease. Contents of polyunsaturated linoleic and linolenic acids increased, and saturated palmitic acid levels tended to decline as the fruit ripened. The increased levels of unsaturated fatty acids closely paralleled increasing expression of PpFAD1 and PpFAD2. The significance of gene expression changes in relation to aroma volatile production is discussed.