Characterization of the Difference between Day and Night Temperatures on the Growth, Photosynthesis, and Metabolite Accumulation of Tea Seedlings.

Currently, the effects of the differences between day and night temperatures (DIFs) on tea plant are poorly understood. In order to investigate the influence of DIFs on the growth, photosynthesis, and metabolite accumulation of tea plants, the plants were cultivated under 5 °C (25/20 °C, light/dark), 10 °C (25/15 °C, light/dark), and 15 °C (25/10 °C, light/dark). The results showed that the growth rate of the new shoots decreased with an increase in the DIFs. There was a downward trend in the photosynthesis among the treatments, as evidenced by the lowest net photosynthetic rate and total chlorophyll at a DIF of 15 °C. In addition, the DIFs significantly affected the primary and secondary metabolites. In particular, the 10 °C DIF treatment contained the lowest levels of soluble sugars, tea polyphenols, and catechins but was abundant in caffeine and amino acids, along with high expression levels of theanine synthetase (TS3) and glutamate synthase (GOGAT). Furthermore, the transcriptome data revealed that the differentially expressed genes were enriched in valine, leucine, and isoleucine degradation, flavone/flavonol biosyntheses, flavonoid biosynthesis, etc. Therefore, we concluded that a DIF of 10 °C was suitable for the protected cultivation of tea plants in terms of the growth and the quality of a favorable flavor of tea, which provided a scientific basis for the protected cultivation of tea seedlings.

Transplantation of Human Umbilical Cord Blood-Derived Cellular Fraction Improves Left Ventricular Function and Remodeling After Myocardial Ischemia/Reperfusion.

Rationale: Human umbilical cord blood (hUCB) contains diverse populations of stem/progenitor cells. Whether hUCB-derived nonhematopoietic cells would induce cardiac repair remains unknown. Objective: To examine whether intramyocardial transplantation of hUCB-derived CD45-Lin- nonhematopoietic cellular fraction after a reperfused myocardial infarction in nonimmunosuppressed rats would improve cardiac function and ameliorate ventricular remodeling. Methods and Results: Nonhematopoietic CD45-Lin- cells were isolated from hUCB. Flow cytometry and quantitative polymerase chain reaction were used to characterize this subpopulation. Age-matched male Fischer 344 rats underwent a 30-minute coronary occlusion followed by reperfusion and 48 hours later received intramyocardial injection of vehicle or hUCB CD45-Lin- cells. After 35 days, compared with vehicle-treated rats, CD45-Lin- cell-treated rats exhibited improved left ventricular function, blunted left ventricular hypertrophy, greater preservation of viable myocardium in the infarct zone, and superior left ventricular remodeling. Mechanistically, hUCB CD45-Lin- cell injection favorably modulated molecular pathways regulating myocardial fibrosis, cardiomyocyte apoptosis, angiogenesis, and inflammation in postinfarct ventricular myocardium. Rare persistent transplanted human cells could be detected at both 4 and 35 days after myocardial infarction. Conclusions: Transplantation of hUCB-derived CD45-Lin- nonhematopoietic cellular subfraction after a reperfused myocardial infarction in nonimmunosuppressed rats ameliorates left ventricular dysfunction and improves remodeling via favorable paracrine modulation of molecular pathways. These findings with human cells in a clinically relevant model of myocardial ischemia/reperfusion in immunocompetent animals may have significant translational implications.Visual Overview: An online visual overview is available for this article.

Weighted Gene Co-Expression Analysis Network-Based Analysis on the Candidate Pathways and Hub Genes in Eggplant Bacterial Wilt-Resistance: A Plant Research Study.

Solanum melongena L. (eggplant) bacterial wilt is a severe soil borne disease. Here, this study aimed to explore the regulation mechanism of eggplant bacterial wilt-resistance by transcriptomics with weighted gene co-expression analysis network (WGCNA). The different expression genes (DEGs) of roots and stems were divided into 21 modules. The module of interest (root: indianred4, stem: coral3) with the highest correlation with the target traits was selected to elucidate resistance genes and pathways. The selected module of roots and stems co-enriched the pathways of MAPK signalling pathway, plant pathogen interaction, and glutathione metabolism. Each top 30 hub genes of the roots and stems co-enriched a large number of receptor kinase genes. A total of 14 interesting resistance-related genes were selected and verified with quantitative polymerase chain reaction (qPCR). The qPCR results were consistent with those of WGCNA. The hub gene of EGP00814 (namely SmRPP13L4) was further functionally verified; SmRPP13L4 positively regulated the resistance of eggplant to bacterial wilt by qPCR and virus-induced gene silencing (VIGS). Our study provides a reference for the interaction between eggplants and bacterial wilt and the breeding of broad-spectrum and specific eggplant varieties that are bacterial wilt-resistant.

Deletion of Interleukin-6 Attenuates Pressure Overload-Induced Left Ventricular Hypertrophy and Dysfunction.

RATIONALE: The role of interleukin (IL)-6 in the pathogenesis of cardiac myocyte hypertrophy remains controversial. OBJECTIVE: To conclusively determine whether IL-6 signaling is essential for the development of pressure overload-induced left ventricular (LV) hypertrophy and to elucidate the underlying molecular pathways. METHODS AND RESULTS: Wild-type and IL-6 knockout (IL-6(-/-)) mice underwent sham surgery or transverse aortic constriction (TAC) to induce pressure overload. Serial echocardiograms and terminal hemodynamic studies revealed attenuated LV hypertrophy and superior preservation of LV function in IL-6(-/-) mice after TAC. The extents of LV remodeling, fibrosis, and apoptosis were reduced in IL-6(-/-) hearts after TAC. Transcriptional and protein assays of myocardial tissue identified Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and signal transducer and activator of transcription 3 (STAT3) activation as important underlying mechanisms during cardiac hypertrophy induced by TAC. The involvement of these pathways in myocyte hypertrophy was verified in isolated cardiac myocytes from wild-type and IL-6(-/-) mice exposed to prohypertrophy agents. Furthermore, overexpression of CaMKII in H9c2 cells increased STAT3 phosphorylation, and exposure of H9c2 cells to IL-6 resulted in STAT3 activation that was attenuated by CaMKII inhibition. Together, these results identify the importance of CaMKII-dependent activation of STAT3 during cardiac myocyte hypertrophy via IL-6 signaling. CONCLUSIONS: Genetic deletion of IL-6 attenuates TAC-induced LV hypertrophy and dysfunction, indicating a critical role played by IL-6 in the pathogenesis of LV hypertrophy in response to pressure overload. CaMKII plays an important role in IL-6-induced STAT3 activation and consequent cardiac myocyte hypertrophy. These findings may have significant therapeutic implications for LV hypertrophy and failure in patients with hypertension.

Formation of controllable hydrophilic/hydrophobic drug delivery systems by electrospinning of vesicles.

Novel multifunctional poly(ethylene oxide) (PEO) nanofibrous membrane, which contains vesicles constructed by mixed surfactant cetyltrimethylammonium bromide (CTAB)/sodium dodecylbenzenesulfonate (SDBS), has been designed as dual drug-delivery system and fabricated via the electrospinning process. 5-FU and paeonolum, which are hydrophilic and hydrophobic anticancer model drugs, can be dissolved in vesicle solution's bond water and lipid bilayer membranes, respectively. The physicochemical properties of the electrospun nanofibrous membrane were systematically studied using scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared (FTIR), and X-ray diffraction (XRD). Drug release behaviors of the electrospun nanofibrous membrane fabricated with different molar ratio of CTAB/SDBS vesicle solution were investigated. The result showed that the releasing amount of hydrophilic drug presented an ascending release manner, while the hydrophobic one showed a descending release behavior with increasing of the molar ratio of CTAB/SDBS. Moreover, the release amount of drugs from drug delivery system can be controlled by the molar ratio of CTAB/SDBS in the vesicle solution easily and conveniently. The distinct properties can be utilized to encapsulate environmental demanding and quantificational materials.

Dual stimuli-responsive self-assembly transition in zwitterionic/anionic surfactant systems.

Temperature and pH responsiveness is important for biological applications in protein reconstitution, gene delivery and controlled drug release. The temperature and pH dual responsive self-assembly transition, vesicle-to-micelle transitions (VMTs) and micelle-to-vesicle transitions (MVTs), in dodecyl sulfonatebetaine (DSB)/sodium bis(2-ethylhexyl) sulfosuccinate (AOT) aqueous solution are investigated. Various experimental techniques including cryogenic transmission electronic microscopy, UV-vis spectroscopy, fluorescence spectroscopy, conductivity, and zeta potential were employed to verify the transformation process. Encapsulation of calcein was further applied in this study. The results showed that the self-assembly transition in DSB/AOT aqueous solution is reversible and can be controlled by temperature and pH. It is anticipated that utilizing simple stimuli methods to realize unique self-assembly behaviour in dilute aqueous solution may offer new possibilities in cancer diagnosis and therapy.

Temperature-induced vesicle to micelle transition in cationic/cationic mixed surfactant systems.

Temperature-induced vesicle to micelle transition (VMT), which has rarely been reported in cationic/cationic mixed surfactant systems, was systemically studied in a didodecyldimethylammonium bromide (DDAB)/dodecyltrimethylammonium chloride (DTAC) aqueous solution. We investigated the effect of temperature on DDAB/DTAC aqueous solutions by means of turbidity, conductivity, cryo-TEM, a UV-vis spectrophotometer, and a steady-state fluorescence spectrometer. It was found that increasing temperature could induce the transformation from the vesicle to the micelle in this cationic/cationic mixed surfactant system. The degree of transformation can be easily controlled by the operation temperature. Additionally, by adjusting the proportion of the mixed cationic/cationic systems and employing cationic surfactants with different chain-lengths, we were able to conclude that the hydrophobic tail length of the surfactant affects the aggregation behavior of cationic/cationic mixed surfactant systems as a function of temperature. It is universal to induce the transformation from the vesicle to the micelle by temperature in cationic/cationic mixed surfactant systems. A possible mechanism for the temperature-induced VMT was proposed based on the experimental results.

CO2-induced micelle to vesicle transition in zwitterionic-anionic surfactant systems.

Study of micelle to vesicle transition (MVT) is of great importance from both theoretical and practical points of view. In this work, we studied the effect of compressed CO2 on the aggregation behavior of zwitterionic-anionic (DSB (dodecyl sulfonatebetaine)-AOT(sodium bis(2-ethylhexyl) sulfosuccinate)) mixed surfactants in aqueous solution by means of direct observation, turbidity, steady-state fluorescence, fluorescence quantum yield, and entrapment quantity of vesicles. Interestingly, all the methods show that compressed CO2 can induce MVT in this zwitterionic-anionic surfactant system. The CO2-induced MVT is reversible and the degree of MVT can be easily tuned by controlling the operation pressure. Further studies show that the pH decrease and dissolution of gas molecules in the surfactant film co-contribute to the MVT, and a possible mechanism for the CO2-induced MVT was proposed based on the experimental results.

Biomolecule-grafted GO enhanced the mechanical and biological properties of 3D printed PLA scaffolds with TPMS porous structure.

Graphene oxide (GO) exhibits excellent mechanical strength and modulus. However, its effectiveness in mechanically reinforcing polymer materials is limited due to issues with interfacial bonding and dispersion arising from differences in the physicochemical properties between GO and polymers. Surface modification using coupling agents is an effective method to improve the bonding problem between polymer and GO, but there may be biocompatibility issues when used in the biomedical field. In this study, the biomolecule L-lysine, was applied to improve the interfacial bonding and dispersion of GO in polylactic acid (PLA) without compromising biocompatibility. The PLA/L-lysine-modified GO (PLA/L-GO) bone scaffold with triply periodic minimal surface (TPMS) structure was prepared using fused deposition modeling (FDM). The FTIR results revealed successful grafting of L-lysine onto GO through the reaction between their -COOH and -NH2 groups. The macroscopic and microscopic morphology characterization indicated that the PLA/L-GO scaffolds exhibited an characteristics of dynamic diameter changes, with good interlayer bonding. It was noteworthy that the L-lysine modification promoted the dispersion of GO and the interfacial bonding with the PLA matrix, as characterized by SEM. As a result, the PLA/0.1L-GO scaffold exhibited higher compressive strength (13.2 MPa) and elastic modulus (226.8 MPa) than PLA/0.1GO. Moreover, PLA/L-GO composite scaffold exhibited superior biomineralization capacity and cell response compared to PLA/GO. In summary, L-lysine not only improved the dispersion and interfacial bonding of GO with PLA, enhancing the mechanical properties, but also improved the biological properties. This study suggests that biomolecules like L-lysine may replace traditional modifiers as an innovative bio-modifier to improve the performance of polymer/inorganic composite biomaterials.

Magnesium Hydroxide as a Versatile Nanofiller for 3D-Printed PLA Bone Scaffolds.

Polylactic acid (PLA) has attracted much attention in bone tissue engineering due to its good biocompatibility and processability, but it still faces problems such as a slow degradation rate, acidic degradation product, weak biomineralization ability, and poor cell response, which limits its wider application in developing bone scaffolds. In this study, Mg(OH)2 nanoparticles were employed as a versatile nanofiller for developing PLA/Mg(OH)2 composite bone scaffolds using fused deposition modeling (FDM) 3D printing technology, and its mechanical, degradation, and biological properties were evaluated. The mechanical tests revealed that a 5 wt% addition of Mg(OH)2 improved the tensile and compressive strengths of the PLA scaffold by 20.50% and 63.97%, respectively. The soaking experiment in phosphate buffered solution (PBS) revealed that the alkaline degradation products of Mg(OH)2 neutralized the acidic degradation products of PLA, thus accelerating the degradation of PLA. The weight loss rate of the PLA/20Mg(OH)2 scaffold (15.40%) was significantly higher than that of PLA (0.15%) on day 28. Meanwhile, the composite scaffolds showed long-term Mg2+ release for more than 28 days. The simulated body fluid (SBF) immersion experiment indicated that Mg(OH)2 promoted the deposition of apatite and improved the biomineralization of PLA scaffolds. The cell culture of bone marrow mesenchymal stem cells (BMSCs) indicated that adding 5 wt% Mg(OH)2 effectively improved cell responses, including adhesion, proliferation, and osteogenic differentiation, due to the release of Mg2+. This study suggests that Mg(OH)2 can simultaneously address various issues related to polymer scaffolds, including degradation, mechanical properties, and cell interaction, having promising applications in tissue engineering.

Forward genetic studies reveal LsAPRR2 as a key gene in regulating the green color of pericarp in bottle gourd (Lagenaria siceraria).

The fruit peel color is an important factor that affects its quality. However, genes involved in regulating pericarp color in bottle gourd (Lagenaria siceraria) have not been explored to date. Genetic analysis of color traits in bottle gourd peel through a genetic population of six generations demonstrated that the green color of peels is inherited as a single gene dominant trait. Combined phenotype-genotype analysis of recombinant plants using BSA-seq mapped the candidate gene to a 22.645 Kb interval at the head end of chromosome 1. We observed that the final interval contained only one gene, LsAPRR2 (HG_GLEAN_10010973). Sequence and spatiotemporal expression analyses of LsAPRR2 unraveled two nonsynonymous mutations (A→G) and (G→C) in the parental CDS sequences. Further, LsAPRR2 expression was higher in all green-skinned bottle gourds (H16) at various stages of fruit development than in white-skinned bottle gourds (H06). Cloning and sequence comparison of the two parental LsAPRR2 promoter regions indicated 11 bases insertion and 8 SNPs mutations in the region -991~-1033, upstream of the start codon in white bottle gourd. Proof of GUS reporting system, Genetic variation in this fragment significantly reduced the expression of LsAPRR2 in the pericarp of white bottle gourd. In addition, we developed a tightly linked (accuracy 93.88%) InDel marker for the promoter variant segment. Overall, the current study provides a theoretical basis for comprehensive elucidation of the regulatory mechanisms underlying the determination of bottle gourd pericarp color. This would further help in the directed molecular design breeding of bottle gourd pericarp.

Prediction of progressive pulmonary fibrosis in patients with anti-synthetase syndrome-associated interstitial lung disease.

OBJECTIVE: Interstitial lung disease (ILD) is a common extramuscular manifestation of the anti-synthetase syndrome (ASS). Patients with ASS-ILD are at risk in developing a progressive fibrosing phenotype despite appropriate treatments. This study investigated the risk factors and the predictive value of multiple risk factors for progressive pulmonary fibrosis (PPF) in patients with ASS-ILD. METHODS: Ninety patients with a diagnosis of ASS and evidence of ILD on high-resolution computed tomography (HRCT) were recruited. Among them, 72 participants completed follow-up for more than 12 months. These patients were further divided into a PPF-ASS group (n = 18) and a non-PPF-ASS group (n = 54). Logistic regression analysis was performed to investigate the risk factors for PPF. The predictive value of the combined risk factors for predicting PPF were analyzed by a ROC curve. RESULTS: The PPF-ASS group had a higher rate of positive non-Jo-1 antibodies, a significantly higher neutrophil-to-lymphocyte ratio (NLR) and serum lactate dehydrogenase (LDH), and a significantly lower PaO2/FiO2 ratio and diffusing capacity for carbon monoxide (DLCO%pred) than the non-PPF-ASS group. In addition, elevated serum Krebs von den Lungen-6 (KL-6) level and reticular opacities were significantly more common, and corticosteroid monotherapy at onset was administered more frequently in the PPF-ASS group. The median duration of follow-up was 37.4 months, survival was poorer in the PPF-ASS group, and the overall survival was 88.9%. Multivariate regression analysis further revealed that positive non-Jo-1 antibodies, NLR, and KL-6 were independent risk factors for PPF. These combined indexes had good accuracy (area under the curve = 0.874) in predicting PPF in patients with ASS-ILD. CONCLUSION: Positive non-Jo-1 antibodies, NLR, and serum KL-6 are independent risk factors for PPF in patients with ASS-ILD. Monitoring these markers can potentially predict PPF in this group of patients. Key Points • Positive non-Jo-1 antibodies, NLR, and serum KL-6 are independent risk factors associated with PPF in patients with ASS-ILD. • Monitoring non-Jo-1 antibodies, NLR, and serum KL-6 can potentially predict PPF in patients with ASS-ILD.

3D printed TPMS structural PLA/GO scaffold: Process parameter optimization, porous structure, mechanical and biological properties.

Bone scaffolds should have good biocompatibility and mechanical and biological properties, which are primarily by the material design, porous structure, and preparation process. In this study, we proposed polylactic acid (PLA) as the base material, graphene oxide (GO) as an enhancing filler, triply periodic minimal surface (TPMS) as a porous structure, and fused deposition modeling (FDM) 3D printing as a preparation technology to develop a TPMS structural PLA/GO scaffold and evaluate their porous structures, mechanical properties, and biological properties towards bone tissue engineering. Firstly, the influence of the FDM 3D printing process parameters on the forming quality and mechanical properties of PLA was studied by orthogonal experimental design, based on which the process parameters were optimized. Then, GO was composited with PLA, and PLA/GO nanocomposites were prepared by FDM. The mechanical tests showed that GO can effectively improve the tensile and compression strength of PLA; only by adding 0.1% GO the tensile and compression modulus was increased by 35.6% and 35.8%, respectively. Then, TPMS structural (Schwarz-P, Gyroid) scaffold models were designed and TPMS structural PLA/0.1%GO nanocomposite scaffolds were prepared by FDM. The compression test showed that the TPMS structural scaffolds had higher compression strength than the Grid structure; This was owing to the fact that the continuous curved structure of TMPS alleviated stress concentration and had a more uniform stress bearing. Moreover, cell culture indicated bone marrow stromal cells (BMSCs) showed better adhesion, proliferation, and osteogenic differentiation behaviors on the TPMS structural scaffolds as the continuous surface structure of TPMS had better connectivity and larger specific surface area. These results suggest that the TPMS structural PLA/GO scaffold has potential application in bone repair. This article suggests the feasibility of co-designing the material, structure, and technology for achieving the good comprehensive performance of polymer bone scaffolds.

A natural biomineral for enhancing the biomineralization and cell response of 3D printed polylactic acid bone scaffolds.

Polylactic acid (PLA) has been extensively used as a bone scaffold material, but it still faces many problems including low biomineralization ability, weak cell response, low mechanical properties, etc. In this study, we proposed to utilize the distinctive physical, chemical and biological properties of a natural biomineral with organic matrix, pearl powder, to enhance the overall performance of PLA bone scaffolds. Porous PLA/pearl composite bone scaffolds were prepared using fused deposition modeling (FDM) 3D printing technology, and their comprehensive performance was investigated. Macro- and micro- morphological observation by the optical camera and scanning electron microscopy (SEM) showed the 3D printed scaffolds have interconnected and ordered periodic porous structures. Phase analysis by X-ray diffraction (XRD) indicated pearl powder was well composited with PLA without impurity formation during the melt extrusion process. The mechanical test results indicated the tensile and compressive strength of PLA/pearl composite scaffolds with 10 % pearl powder content yielded the highest values, which were 15.5 % and 21.8% greater than pure PLA, respectively. The water contact angle and water absorption tests indicated that PLA/pearl showed better hydrophilicity than PLA due to the presence of polar groups in the organic matrix of the pearl powder. The results of the simulated body fluid (SBF) soaking revealed that the addition of pearl powder effectively enhanced the formation and deposition of apatite, which was attributed to the release of Ca2+ from the dissolution of pearl powder. The cell culture of bone marrow mesenchymal stem cells (BMSCs) indicated that PLA/pearl scaffolds showed better cell proliferation and osteogenic differentiation than PLA due to the stimulation of the biological organic matrix in pearl powder. These outcomes signify the potential of pearl powder as a natural biomineral containing bio-signal factors to improve the mechanical and biological properties of polymers for better bone tissue engineering application.

Proteomic analysis of the effects of exogenous calcium on hypoxic-responsive proteins in cucumber roots.

BACKGROUND: Hypoxia acts as a plant stress factor, particularly in cucumbers plants under hydroponic culture. Calcium is involved in stress signal transmission and in the growth of plants. To determine the effect of exogenous calcium on hypoxic-responsive proteins in cucumber (Cucumis sativus L. cv. Jinchun No.2) roots, proteomic analysis was performed using two-dimensional electrophoresis (2-DE) and mass spectrometry. RESULTS: Cucumber roots were used to analyze the influence of hypoxia on plants. The expressions of 38 protein spots corresponding to enzymes were shown to change in response to hypoxia. Of these, 30 spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF MS analysis). The proteins were categorized according to functional groups, including glycolysis, the tricarboxylic acid (TCA) cycle, fermentative metabolism, nitrogen metabolism, energy metabolism, protein synthesis and defense against stress. Exogenous calcium appeared to alleviate hypoxic stress via these metabolic and physiological systems. Western blotting was used to analyze the accumulation of alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC); calcium further increased the expression of ADH and PDC under hypoxia. In addition, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to assess the transcript levels of differentially expressed proteins. CONCLUSIONS: Exogenous calcium enhanced the expression of enzymes involved in glycolysis, the TCA cycle, fermentative metabolism, nitrogen metabolism, and reactive oxygen species (ROS) defense in plants under hypoxia. Calcium appears to induce hypoxic tolerance of cucumber seedlings. These phenomena have prompted us to further investigate the mechanisms by which cucumbers respond to exogenous calcium under hypoxia.

[The expression of thrombospondin-1 in serum and pulmonary arterioles of hypoxic pulmonary hypertension rats].

OBJECTIVE: To investigate the expression of thrombospondin-1 (TSP-1) in serum and pulmonary arterioles of rats with hypoxic pulmonary hypertension. METHODS: Twenty male Wistar rats were divided into two groups and exposed to air and isobaric hypoxia for 3 weeks respectively. The mean pulmonary artery pressure (mPAP) was measured by right cardiac catheterization. The rates of wall thickness/external diameter (WT%) and wall area/total vascular area (WA%) were calculated. The TSP-1 level in serum was measured by enzyme-linked immunosorbent assay. TSP-1 mRNA expression in lung tissue was evaluated by quantitative PCR. RESULTS: The pulmonary artery pressure increased in the hypoxia exposed rats. The chronic hypoxia also elicited the thicking of the wall and the narrowing of the lumen of pulmonary arterioles. It led to the increases of pulmonary artery pressure, the index of right ventricular hypertrophy [RV/(LV+S)], WA% and WT% compared to the controls [mPAP:(2.86 +/- 0.39) kPa vs. (1.35 +/- 40.28) kPa; RV/(LV+ S): (43.53 +/- 3.38)% vs. (23.68 +/- 3.48)%; WT%: (35.24 +/- 11.20)% vs. (23.63 +/- 9.74)%; WA%: (55.09 +/- 12.38)% vs. (41.62 +/- 12.83)% respectively, P<0.05]. In hypoxic group, the expression of TSP-1 mRNA in the lung was significantly up-regulated, the expression level of TSP-1 in serum was higher than that in control group (P<0.01). Linear correlation analysis showed that TSP-1 mRNA was positively associated with WT%, WA% and mPAP (r= 0.748, 0.686, 0.942 respectively, P<0.05). CONCLUSION: The TSP-1 may play an important role in the pathogenesis process of hypoxic pulmonary vascular remodeling and pulmonary hypertension.

[Expression of thrombospondin-1 in the lung of hypoxic pulmonary hypertension rats].

OBJECTIVE: To investigate the expression of thrombospondin-1(TSP-1) in the lung of hypoxia-induced pulmonary hypertension rats. METHODS: Thirty male Wistar rats were divided into two groups, pulmonary hypertension group and control group. The mice in experimental group were exposed to isobaric hypoxia for 3 weeks, and those in control group were exposed to air. The pulmonary artery pressure was measured by right cardiac catheterization. The expression of TSP-1 and TGF-beta1 in the lungs of rats were measured by immunohistochemical staining. The histological sections of the lungs were examined using a computerized image analyzer. RESULTS: After the induction of hypoxia for 3 weeks, the rats had pulmonary artery pressure increased with the thickening of the wall and the narrowing of the lumen of pulmonary arterioles. In the experimental group, the mean pulmonary artery pressure (mPAP) was (2.86 +/- 0.39) kPa, the index of right ventricular hypertrophy RV/(LV+S) was (43.53 +/- 3.38)%, the ratio of vascular wall thickness/vascular external diameter (WA%) was (55.09 +/- 12.38)%, and the ratio of vascular wall area/total vascular area (WT%) was (35.24 +/- 11.2)%, which all were significantly increased in comparison with those of control group [mPAP (1.35 +/- 0.28) kPa, RV/(LV+S) (23.68 +/- 3.48)%, WT% (23.63 +/- 9.74)%, WA% (41.62 +/- 12.83)%, respectively. P < 0.05). The positive staining of TSP-1 (1.32 +/- 0.04 vs. 0.96 +/- 0.03) and TGF-beta1 (1.38 +/- 0.05 vs. 1.04 +/- 0.04) in the wall of pulmonary arteriole of the rats exposed to hypoxia were significantly stronger than those of control rats (P < 0.01). CONCLUSION: The expression of TSP-1 appears to be increased in hypoxic pulmonary hypertension rats, which may contribute to the pathogenesis of hypoxic pulmonary vascular remodeling.

[Recombinant human interleukin 35 (rhIL-35) alleviates the damage of coronary artery endothelial cells in Kawasaki disease by inhibiting NF-κB pathway].

Objective To investigate the protective effect and mechanism of recombinant human interleukin 35(rhIL-35) on coronary artery injury in Kawasaki disease (KD). Methods Human coronary artery endothelial cells (HCAECs) were cultured in vitro to establish KD vascular model. Tumor necrosis factor α(TNF-α) and the serum of KD patients stimulated HCAECs were used to mimic the local inflammatory lesions of KD. The cells were divided into control group, TNF-α and KD serum stimulation group, (25, 50) ng/mL rhIL-35 treatment group. Cell proliferation was detected by CCK-8 assay; mRNA levels of IL-1ß, IL-6, IL-17A and zonula occludens-1(ZO-1) of HCAECs were detected by real-time quantitative PCR; IL-35 expression in plasma and IL-1ß, IL-6 and IL-17A content in HCAEC supernatant were tested by ELISA; Western blot was performed to detect the expression of nuclear factor κB p65 (NF-κB p65) and ZO-1. Results TNF-α and KD serum inhibited the proliferation of HCAECs, while rhIL-35 significantly reversed the above effects. RhIL-35 significantly down-regulated the expression of IL-1ß, IL-6 and IL-17A after preconditioning HCAECs. Compared with TNF-α and KD serum stimulation group, rhIL-35 pretreated cells could significantly increase ZO-1 protein expression and inhibit NF-κB p65 expression. Conclusions rhIL-35 can alleviate the damage of KD coronary artery endothelial cells by inhibiting NF-κB pathway.

Tyrosine sulfation in N-terminal domain of human C5a receptor is necessary for binding of chemotaxis inhibitory protein of Staphylococcus aureus.

AIM: Staphylococcus aureus evades host defense through releasing several virulence proteins, such as chemotaxis inhibitory protein of staphylococcus aureus (CHIPS). It has been shown that extracellular N terminus of C5a receptor (C5aR) forms the binding domain for CHIPS, and tyrosine sulfation is emerging as a key factor in determining protein-protein interaction. The aim of this study was to evaluate the role of tyrosine sulfation of N-terminal of C5aR in its binding with CHIPS. METHODS: Expression plasmids encoding C5aR and its mutants were prepared using PCR and site-directed mutagenesis and were used to transfect HEK 293T cells using calcium phosphate. Recombinant CHIPS protein was purified. Western blotting was used to examine the binding efficiency of CHIPS to C5aR or its mutants. RESULTS: CHIPS exclusively binds to C5aR, but not to C5L2 or C3aR. A nonspecific sulfation inhibitor, sodium chlorate (50 nmol/L), diminishes the binding ability of C5aR with CHIPS. Blocking sulfation by mutation of tyrosine to phenylalanine at positions 11 and 14 of C5aR N terminus, which blocked sulfation, completely abrogates CHIPS binding. When tyrosine 14 alone was mutated to phenylalanine, the binding efficiency of recombinant CHIPS was substantially decreased. CONCLUSION: The results demonstrate a structural basis of C5aR-CHIPS association, in which tyrosine sulfation of N-terminal C5aR plays an important role. Our data may have potential significance in development of novel drugs for therapeutic intervention.

Mechanochemical synthesis of Fe2O3/Zn-Al layered double hydroxide based on red mud.

In this work, using industrial waste red mud (RM) as main starting material, a simple method of mechanochemical synthesis (MCS) was introduced as a green approach to synthesize heterogeneous Fe2O3/Zn-Al layered double hydroxide (F/ZA-LDH), which could be used as a new low-cost catalyst for photo-Fenton reaction. The optimum preparation conditions of F/ZA-LDH were as follows: mass ratio of Zn(NO3)2·6H2O to RM (mZn:mRM) 2:1, dry milling time 6 h, H2O dosage 2 mL, ball-to-powder mass ratio 50:1, and milling speed 250 rpm. The effects of the synthesis conditions on the crystal structure and catalytic activity of F/ZA-LDH were analyzed. The F/ZA-LDH was characterized by XRD, TG, XPS, SEM, (HR)TEM. The characterization results showed the composite had a crystallized hydrotalcite-like structure, and the crystalline phases in the optimum F/ZA-LDH were Fe2O3 and Zn-Al LDH. A hetero-interfaces between Fe2O3 and Zn-Al LDH existed in the synthesized Fe2O3/Zn-Al LDH composite. Furthermore, the possible mechanism for F/ZA-LDH formation in the MCS process was proposed. Overall, our results provide a systematic understanding of the preparation of LDH composite through MCS using RM as main material, and our findings help to develop green technology for reusing RM.