C1q/tumour necrosis factor-related protein-9 aggravates lipopolysaccharide-induced inflammation via promoting NLRP3 inflammasome activation.

The NLRP3 inflammasome plays a vital role in inflammation by increasing the maturation of interleukin-1ß (IL-1ß) and promoting pyroptosis. Given that C1q/tumour necrosis factor-related protein-9 (CTRP9) has been shown to be involved in diverse inflammatory diseases, we sought to assess the underlying impact of CTRP9 on NLRP3 inflammasome activation. In vitro, macrophages isolated from murine peritonea were stimulated with exogenous CTRP9, followed by lipopolysaccharide (LPS) and adenosine 5'-triphosphate (ATP). We demonstrated that CTRP9 markedly augmented the activation of the NLRP3 inflammasome, as shown by increased mature IL-1ß secretion, triggering ASC speck formation and promoting pyroptosis. Mechanistically, CTRP9 increased the levels of NADPH oxidase 2 (NOX2)-derived reactive oxygen species (ROS). Suppressing ROS with N-acetylcysteine (NAC) or interfering with NOX2 by small interfering RNA weakened the promoting effect of CTRP9 on the NLRP3 inflammasome. Furthermore, NLRP3 inflammasome activation, pyroptosis and secretion of mature IL-1ß were significantly decreased in macrophages from CTRP9-KO mice compared to those from WT mice with the same treatment. In vivo, we established a sepsis model by intraperitoneal injection of LPS into WT and CTRP9-KO mice. CTRP9 knockout improved the survival rates of the septic mice and attenuated NLRP3 inflammasome-mediated inflammation. In conclusion, our study indicates that CTRP9 aggravates LPS-induced inflammation by promoting NLRP3 inflammasome activation via the NOX2/ROS pathway. CTRP9 could be a promising target for NLRP3 inflammasome-driven inflammatory diseases.

Anti-osteoporosis effect and purification of peptides with high calcium-binding capacity from walnut protein hydrolysates.

The walnut protein hydrolysate (WPH) was prepared via simulated gastrointestinal digestion. The degree of hydrolysis (DH), amino acid composition, and relative molecular weight distribution of WPH were analyzed. The results showed that the DH of WPH was 11.6%, WPH was rich in Glu and Pro, and the relative average molecular weight of 572 Da accounted for 59.78%. The effects of WPH on osteoporosis were evaluated using a model of retinoic acid-induced osteoporosis rat. Treatment with WPH effectively increased the serum calcium and phosphorus contents, alleviated calcium loss, and reduced tartrate-resistant acid phosphate and alkaline phosphatase activities and bone gla protein content. WPH treatment significantly improved the biomechanical properties of the bone and increased the value of bone mineral density. In addition, WPH treatment improved the bone microstructure. WPH was isolated and purified by Sephadex G-25 gel filtration chromatography and semi-preparative reversed-phase high-performance liquid chromatography. A fraction with high calcium-binding activity was obtained and 15 peptides were identified.

Globular CTRP9 protects cardiomyocytes from palmitic acid-induced oxidative stress by enhancing autophagic flux.

BACKGROUND: Oxidative stress in cardiac myocytes is an important pathogenesis of cardiac lipotoxicity. Autophagy is a cellular self-digestion process that can selectively remove damaged organelles under oxidative stress, and thus presents a potential therapeutic target against cardiac lipotoxicity. Globular CTRP9 (gCTRP9) is a newly identified adiponectin paralog with established metabolic regulatory properties. The aim of this work is to investigate whether autophagy participates the protection effects of gCTRP9 in neonatal rat cardiac myocytes (NRCMs) under oxidative stress and the underlying mechanism. RESULTS: NRCMs were treated with PA of various concentrations for indicated time period. Our results showed that PA enhanced intracellular ROS accumulation, decreased mitochondrial membrane potential (Δψm) and increased activation of caspases 3. These changes suggested lipotoxicity due to excessive PA. In addition, PA was observed to impair autophagic flux in NRCMs and impaired autophagosome clearance induced by PA contributes to cardiomyocyte death. Besides, we found that gCTRP9 increased the ratio of LC3II/I and the expression of ATG5 which was vital to the formation of autophagosomes and decreased the level of P62, suggesting enhanced autophagic flux in the absence or presence of PA. The result was further confirmed by the methods of infection with LC3-mRFP-GFP lentivirus and blockage of autophagosome-lysosome fusion by BafA1. Moreover, gCTRP9 reestablished the loss of mitochondrial membrane potential, suppressed ROS generation, and reduced PA -induced myocyte death. However, the protective effect of gCTRP9 on the cardiac lipotoxicity was partly abolished by blockade of autophagy by autophagy-related 5 (ATG5) siRNA, indicating that the effect of gCTRP9 on cell survival is critically mediated through regulation of autophagy. CONCLUSION: Autophagy induction by gCTRP9 could be utilized as a potential therapeutic strategy against oxidative stress-mediated damage in cardiomyocytes.

Physical Properties of Composite Films from Tilapia Skin Collagen with Pachyrhizus Starch and Rambutan Peel Phenolics.

Different composite films composed of tilapia skin collagen (TSC) with Pachyrhizus starch (PS) or rambutan peel phenolics (RPP) were prepared, and the physical properties of these films were determined. The effects of PS and RPP on TSC films were investigated, and our results indicated that PS and RPP could improve the physical properties of TSC films. Opacity and film thickness showed an enhanced trend with increasing PS and RPP contents in TSC films, whereas solubility in water, elongation-at-break (EAB), and water vapor permeability (WVP) showed declining trends. TSC film with 10% PS and 0.5% RPP had the highest tensile strength, and the tensile strength dropped drastically when the content of PS and RPP increased. The light transmittances of the films could decrease with the incorporation of PS and RPP. Differential scanning calorimetry (DSC) demonstrated that the addition of PS and RPP improved the thermal stability of TSC films. In addition, X-ray diffraction indicated that the crystallinity of the films decreased and the amorphous structure of the films tended to become more complex with the addition of PS and RPP. As shown by fourier transform infrared spectroscopy (FTIR) analysis, PS and RPP can strongly interact with TSC, resulting in a modification of its structure. Scanning electron microscope (SEM) analysis showed that there was a good compatibility between TSC, PS, and RPP. The results indicated that TSC film incorporated with 10% PS and 0.5% RPP was an effective method for improve the physical properties of the film. TSC-PS-RPP composite films can be used not only in biomedical applications, but also as active food packaging materials.

Prospective analysis of glycemic variability in patients with severe traumatic brain injury: modified Leuven's adjustment process versus conventional adjustment process.

Objective This study was performed to evaluate the effect of two different methods of controlling glycemic variability (GV) in patients with severe traumatic brain injury (STBI) undergoing surgery. Methods Patients with STBI were randomly grouped into a conventional adjustment process (CAP) group and modified Leuven's adjustment process (mLAP) group. Each group included 50 patients. Blood glucose levels were continuously monitored and data were recorded and analyzed. Results The mean blood glucose level was stable in both groups for 5 days postoperatively with no significant difference. The standard deviation of the blood glucose level, mean amplitude of glycemic excursions, and glycemic lability index were significantly higher in the CAP than mLAP group for the first 2 days. In the final 3 days, no significant differences were observed between the two groups. The incidence of hypoglycemia was significantly higher in the CAP than mLAP group on the first day. This value gradually declined during the following 4 days, but the difference between the two groups was not significant. Conclusion The mLAP produced more favorable results than the CAP for GV control in the early stage after surgery for STBI.

The Influence of Sedation Level Guided by Bispectral Index on Therapeutic Effects for Patients with Severe Traumatic Brain Injury.

BACKGROUND: Sedation therapy is vital for treating severe traumatic brain injury (TBI). Yet, types of sedation assessment tools and sedation levels that are suitable for sedation treatment have not been investigated. OBJECTIVE: To investigate the influence of different sedation levels guided by the Bispectral Index (BIS) on the therapeutic effects for severe TBI. METHODS: According to inclusion, exclusion, and rejection criteria, 35 patients were prospectively included and divided into Richmond Agitation Sedation Scale (RASS), BIS(I), and BIS(II) groups. The RASS group was controlled the level of sedation to within -2 or -3, and the BIS(I) and (II) groups within the range of 40-50 and 50-60, respectively. In addition to clinical data, RASS, BIS, and intracranial pressure (ICP) values were collected. RASS and ICP variability were introduced to investigate the different of sedative control effect with or without BIS monitor, and the control effect of ICP between different sedation levels. Statistical analysis was performed to estimate the effectiveness of different sedation levels guided by BIS in sedation treatment within 72 hours. RESULTS: There were no significant differences in demographics among the 3 groups. RASS variability of the BIS(I) and (II) groups was significantly lower than that of the RASS group (P < 0.05), and in the BIS(I) group it was insignificantly lower than in the BIS(II) group. The ICP of the BIS(I) and (II) groups declined to <13.5 mm Hg significantly earlier than that of the RASS group (P < 0.05), and the difference between BIS(I) and (II) was insignificant. ICP variability of RASS group was higher than those of the BIS(I) and (II) groups (P < 0.05), and ICP variability of the BIS(I) group was significantly lower than that of the BIS(II) group (P < 0.05). Differences in days in the neurosurgery intensive care unit and outcomes among the 3 groups were insignificant. CONCLUSIONS: BIS is more reliable than RASS for maintaining a stable sedation status and ICP. Deeper sedation levels (BIS 40-50) cause ICP to decrease more quickly, with lower ICP variability.