Pain Intensity and Satisfaction of Pain Relief in Discharged Cancer Patients: A Large Sample Study in China.

BACKGROUND: Many studies have focused on the quality of pain management in hospitalized patients with cancer pain, while what happens after discharge remains unclear. AIM: The purpose of this study was to investigate the pain intensity and satisfaction of pain relief among a large sample of Chinese patients with cancer pain after discharge. DESIGN: Cross-sectional, descriptive, correlational research. SETTINGS AND SAMPLE: ABOUT: 1,013 patients were recruited in a tertiary cancer hospital, and their residence addresses were distributed in 6 geographical regions, including 26 provinces, municipalities, and autonomous regions. METHODS: The 1,013 patients with cancer pain were discharged from the wards of a national cancer hospital in China from July 2020 to October 2021. A nurse in the pain clinic followed the patients based on a whole-process information system and collected the data after the cancer pain patients were discharged. The study methods followed the STROBE guidelines. RESULTS: The average age of 1,013 discharged patients was 61.30 (±12.56) years. Moderate and severe background pain (BGP) was reported in 749 patients (73.94%), and more than 3 instances of breakthrough pain (BTP) in the past 24 hours were reported in 541 patients (53.41%). More severe BGP was associated with more frequent BTP (p < .01). In addition, there were 572 patients (56.47%) whose satisfaction with pain relief was lower than 70%. More severe BGP was associated with a lower satisfaction degree (r = -0.796, p < .01). CONCLUSIONS: Pain among discharged Chinese patients with cancer is poorly managed, and there is a low degree of satisfaction with pain relief. Nurses can do more work to assist cancer patients in managing pain more effectively by ensuring they have a plan to report and manage pain after discharge.

p21-activated kinase 4 inhibition protects against liver ischemia/reperfusion injury: Role of nuclear factor erythroid 2-related factor 2 phosphorylation.

BACKGROUND AND AIMS: p21-activated kinase 4 (PAK4), an oncogenic protein, has emerged as a promising target for anticancer drug development. Its role in oxidative stress conditions, however, remains elusive. We investigated the effects of PAK4 signaling on hepatic ischemia/reperfusion (I/R) injury. APPROACH AND RESULTS: Hepatocyte- and myeloid-specific Pak4 knockout (KO) mice and their littermate controls were subjected to a partial hepatic I/R (HIR) injury. We manipulated the catalytic activity of PAK4, either through genetic engineering (gene knockout, overexpression of wild-type [WT] or dominant-negative kinase) or pharmacological inhibitor, coupled with a readout of nuclear factor erythroid 2-related factor 2 (Nrf2) activity, to test the potential function of PAK4 on HIR injury. PAK4 expression was markedly up-regulated in liver during HIR injury in mice and humans. Deletion of PAK4 in hepatocytes, but not in myeloid cells, ameliorated liver damages, as demonstrated in the decrease in hepatocellular necrosis and inflammatory responses. Conversely, the forced expression of WT PAK4 aggravated the pathological changes. PAK4 directly phosphorylated Nrf2 at T369, and it led to its nuclear export and proteasomal degradation, all of which impaired antioxidant responses in hepatocytes. Nrf2 silencing in liver abolished the protective effects of PAK4 deficiency. A PAK4 inhibitor protected mice from HIR injury. CONCLUSIONS: PAK4 phosphorylates Nrf2 and suppresses its transcriptional activity. Genetic or pharmacological suppression of PAK4 alleviates HIR injury. Thus, PAK4 inhibition may represent a promising intervention against I/R-induced liver injury.

Research on Bathymetric Inversion Capability of Different Multispectral Remote Sensing Images in Seaports.

In recent years, remote sensing has become an indispensable supplementary method for determining water depth in the seaports. At present, many scholars use multi-spectral satellite data to invert the water depth of the seaports, but how to select the appropriate satellite data in the seaports area is worth exploring. In this article, the differences in the retrieving ability between domestic and foreign multispectral images are compared, through building the random forest model and the band ratio model, which use different multispectral images to conduct retrieving water depth in Nanshan Port in conjunction with the WBMS multi-beam sounding system. The band ratio model and random forest model are chosen for water depth exploration, remote sensing images use GF-6, GF-2, Sentinel-2B, and Landsat 8 OLI data, which are all popular and easily accessible. The final experiment results from the constant adjustment of the model parameter show that the domestic series of GF-6 images performed the best in this experiment. The Root Mean Square Error (RMSE) and Mean Relative Error (MRE) of the random forest model are only 1.202 and 0.187, respectively. Simultaneously, it is discovered that the 'Red Edge' band of GF-6 is also very helpful in improving the accuracy of water depth inversion, which is rarely mentioned in previous studies. To some extent, the preceding studies demonstrate that it is possible to investigate water depth using common multispectral remote sensing images. In the case of some bathymetry inversion models or in some waters, the aforementioned study demonstrates that it is possible to examine the water depth using domestic remote sensing images that are superior to foreign multispectral images in terms of bathymetry inversion ability.

[A single-center retrospective analysis of 46 children with aerophagia].

OBJECTIVE: To study the clinical features of aerophagia in children. MEYJODS: A retrospective analysis was performed on the medical data of 46 children with aerophagia who were diagnosed and treated in Children's Hospital Affiliated to Nanjing Medical University from October 2011 to September 2019. RESULTS: Among these 46 children, 15 (33%) had Tourette syndrome. Abdominal distension was the most common symptom and was observed in 45 children (98%). The 24-hour esophageal multichannel intraluminal impedance monitoring showed a mean number of 341 times of air swallowing and a mean number of 212 times of gas reflux, and 95% of gas refluxes occurred in the upright body position. Compared with those without Tourette syndrome, the children with Tourette syndrome had a significantly higher incidence rate of air swallowing symptoms (67% vs 6%, P<0.001), but there were no significant differences in other symptoms and the results of 24-hour esophageal impedance. Dietary adjustment, psycho-behavioral therapy, and drug intervention significantly improved the scores of clinical symptoms and quality of life, among which psycho-behavioral therapy was an important intervention measure. CONCLUSIONS: Some children with aerophagia may have Tourette syndrome, and such children are more likely to have air swallowing symptoms. Psycho-behavioral therapy is one of the most important treatment methods, and children with aerophagia tend to have a good prognosis after treatment.

Elevated CO2 concentration induces photosynthetic down-regulation with changes in leaf structure, non-structural carbohydrates and nitrogen content of soybean.

BACKGROUND: Understanding the mechanisms of crops in response to elevated CO2 concentrations is pivotal to estimating the impacts of climate change on the global agricultural production. Based on earlier results of the "doubling-CO2 concentration" experiments, many current climate models may overestimate the CO2 fertilization effect on crops, and meanwhile, underestimate the potential impacts of future climate change on global agriculture ecosystem when the atmospheric CO2 concentration goes beyond the optimal levels for crop growth. RESULTS: This study examined the photosynthetic response of soybean (Glycine max (L.) Merr.) to elevated CO2 concentration associated with changes in leaf structure, non-structural carbohydrates and nitrogen content with environmental growth chambers where the CO2 concentration was controlled at 400, 600, 800, 1000, 1200, 1400, 1600 ppm. We found CO2-induced down-regulation of leaf photosynthesis as evidenced by the consistently declined leaf net photosynthetic rate (An) with elevated CO2 concentrations. This down-regulation of leaf photosynthesis was evident in biochemical and photochemical processes since the maximum carboxylation rate (Vcmax) and the maximum electron transport rate (Jmax) were dramatically decreased at higher CO2 concentrations exceeding their optimal values of about 600 ppm and 400 ppm, respectively. Moreover, the down-regulation of leaf photosynthesis at high CO2 concentration was partially attributed to the reduced stomatal conductance (Gs) as demonstrated by the declines in stomatal density and stomatal area as well as the changes in the spatial distribution pattern of stomata. In addition, the smaller total mesophyll size (palisade and spongy tissues) and the lower nitrogen availability may also contribute to the down-regulation of leaf photosynthesis when soybean subjected to high CO2 concentration environment. CONCLUSIONS: Down-regulation of leaf photosynthesis associated with the changes in stomatal traits, mesophyll tissue size, non-structural carbohydrates, and nitrogen availability of soybean in response to future high atmospheric CO2 concentration and climate change.

The optimal CO2 concentrations for the growth of three perennial grass species.

BACKGROUND: Grasslands are one of the most representative vegetation types accounting for about 20% of the global land area and thus the response of grasslands to climate change plays a pivotal role in terrestrial carbon balance. However, many current climate change models, based on earlier results of the doubling-CO2 experiments, may overestimate the CO2 fertilization effect, and as a result underestimate the potentially effects of future climate change on global grasslands when the atmospheric CO2 concentration goes beyond the optimal level. Here, we examined the optimal atmospheric CO2 concentration effect on CO2 fertilization and further on the growth of three perennial grasses in growth chambers with the CO2 concentration at 400, 600, 800, 1000, and 1200 ppm, respectively. RESULTS: All three perennial grasses featured an apparent optimal CO2 concentration for growth. Initial increases in atmospheric CO2 concentration substantially enhanced the plant biomass of the three perennial grasses through the CO2 fertilization effect, but this CO2 fertilization effect was dramatically compromised with further rising atmospheric CO2 concentration beyond the optimum. The optimal CO2 concentration for the growth of tall fescue was lower than those of perennial ryegrass and Kentucky bluegrass, and thus the CO2 fertilization effect on tall fescue disappeared earlier than the other two species. By contrast, the weaker CO2 fertilization effect on the growth of perennial ryegrass and Kentucky bluegrass was sustained for a longer period due to their higher optimal CO2 concentrations than tall fescue. The limiting effects of excessively high CO2 concentrations may not only associate with changes in the biochemical and photochemical processes of photosynthesis, but also attribute to the declines in stomatal conductance and nitrogen availability. CONCLUSIONS: In this study, we found apparent differences in the optimal CO2 concentrations for the growth of three grasses. These results suggest that the growth of different types of grasses may respond differently to future elevated CO2 concentrations through the CO2 fertilization effect, and thus potentially alter the community composition and structure of grasslands. Meanwhile, our results may also be helpful for improving current process-based ecological models to more accurately predict the structure and function of grassland ecosystems under future rising atmospheric CO2 concentration and climate change scenarios.

An aptamer assay using rolling circle amplification coupled with thrombin catalysis for protein detection.

We describe a sensitive aptamer-based sandwich assay for protein detection on microplate by using rolling circle amplification (RCA) coupled with thrombin catalysis. This assay takes advantage of RCA generating long DNA oligonucleotides with repeat thrombin-binding aptamer sequence, specific aptamer affinity binding to achieve multiple thrombin labeling, and enzyme activity of thrombin for signal generation. Protein target is specifically captured by antibody-coated microplate. Then, an oligonucleotide containing an aptamer for protein and a primer sequence is added to form a typical sandwich structure. Following a template encoded with complementary sequence of aptamer for thrombin, RCA reaction extends the primer sequence into a long oligonucleotide. Many thrombin molecules bind with the RCA product. Thrombin catalyzes the conversion of its chromogenic or fluorogenic peptide substrates into detectable products for final quantification of protein targets. We applied this strategy to the detection of a model protein target, platelet-derived growth factor-BB (PDGF-BB). Due to double signal amplifications from RCA and thrombin catalysis, this assay enabled the detection of PDGF-BB as low as 3.1 pM when a fluorogenic peptide substrate was used. This assay provides a new way for signal generation in RCA-involved assay through direct thrombin labeling, circumventing time-consuming preparation of enzyme-conjugate and affinity probes. This method has promise for a variety of analytical applications.

Leaf photosynthetic pigment as a predictor of leaf maximum carboxylation rate in a farmland ecosystem.

The leaf maximum rate of carboxylation (Vcmax) is a key parameter of plant photosynthetic capacity. The accurate estimation of Vcmax is crucial for correctly predicting the carbon flux in the terrestrial carbon cycle. Vcmax is correlated with plant traits including leaf nitrogen (Narea) and leaf photosynthetic pigments. Proxies for leaf chlorophyll (Chlarea) and carotenoid contents (Cararea) need to be explored in different ecosystems. In this study, we evaluated the relationship between leaf maximum rate of carboxylation (scaled to 25°C; Vcmax25) and both leaf Narea and photosynthetic pigments (Chlarea and Cararea) in winter wheat in a farmland ecosystem. Our results showed that Vcmax25 followed the same trends as leaf Chlarea. However, leaf Narea showed smaller dynamic changes before the flowering stage, and there were smaller seasonal variations in leaf Cararea. The correlation between leaf Vcmax25 and leaf Chlarea was the strongest, followed by leaf Cararea and leaf Narea (R2 = 0.69, R2 = 0.47 and R2 = 0.36, respectively). The random forest regression analysis also showed that leaf Chlarea and leaf Cararea were more important than leaf Narea for Vcmax25. The correlation between leaf Vcmax25 and Narea can be weaker since nitrogen allocation is dynamic. The estimation accuracy of the Vcmax25 model based on Narea, Chlarea, and Cararea (R2 = 0.75) was only 0.05 higher than that of the Vcmax25 model based on Chlarea and Cararea (R2 = 0.70). However, the estimation accuracy of the Vcmax25 model based on Chlarea and Cararea (R2 = 0.70) was 0.34 higher than that of the Vcmax25 model based on Narea (R2 = 0.36). These results highlight that leaf photosynthetic pigments can be a predictor for estimating Vcmax25, expanding a new way to estimate spatially continuous Vcmax25 on a regional scale, and to improve model simulation accuracy.

Effects of elevated carbon dioxide on plant growth and leaf photosynthesis of annual ryegrass along a phosphorus deficiency gradient.

Introduction: Soil phosphorus (P) deficiency limits plant growth and productivity in grassland ecosystems and may moderate the growth-promoting effects of "carbon dioxide (CO2) fertilization effect". Methods: To evaluate the interactive effects of these two factors on the growth and physiology for annual ryegrass (Lolium multiflorum Lam.), plants were grown in controlled growth chambers with a range of P supply (0.004, 0.012, 0.02, 0.06, 0.1 and 0.5 mM) under two levels of CO2 (400 and 800 µmol mol-1, respectively). Results: Elevated [CO2] dramatically increased the aboveground biomass and net photosynthetic rates of annual ryegrass by 14.5% and 25.3% under sufficient P supply (0.5 mM), respectively, whereas decreased the belowground biomass and net photosynthetic rates under lower P supply of P0.004, P0.02, and P0.06. Two-way ANOVA results showed that CO2 × P (p < 0.001) significantly affected stomatal traits, leaf photosynthesis and biomass. The stimulation of growth and photosynthesis by elevated CO2 concentration (e[CO2]) was reduced or highly suppressed, indicating that the sensitivity of annual ryegrass to P deficiency was enhanced under e[CO2]. Discussion: These results indicated that P limitation may offset the positive effects of e[CO2] on plant growth by altering stomatal traits, leaf photochemical processes and biochemical composition in annual ryegrass.

Effects of elevated CO2 concentration and experimental warming on morphological, physiological, and biochemical responses of winter wheat under soil water deficiency.

Global climate change and freshwater scarcity have become two major environmental issues that constrain the sustainable development of the world economy. Climate warming caused by increasing atmospheric CO2 concentration can change global/regional rainfall patterns, leading to uneven global seasonal precipitation distribution and frequent regional extreme drought events, resulting in a drastic reduction of available water resources during the critical crop reproduction period, thus causing many important food-producing regions to face severe water deficiency problems. Understanding the potential processes and mechanisms of crops in response to elevated CO2 concentration and temperature under soil water deficiency may further shed lights on the potential risks of climate change on the primary productivity and grain yield of agriculture. We examined the effects of elevated CO2 concentration (e[CO2]) and temperature (experimental warming) on plant biomass and leaf area, stomatal morphology and distribution, leaf gas exchange and mesophyll anatomy, rubisco activity and gene expression level of winter wheat grown at soil water deficiency with environmental growth chambers. We found that e[CO2] × water × warming sharply reduced plant biomass by 57% and leaf photosynthesis (P n) 50%, although elevated [CO2] could alleviated the stress from water × warming at the amount of gene expression in RbcL3 (128%) and RbcS2 (215%). At ambient [CO2], the combined stress of warming and water deficiency resulted in a significant decrease in biomass (52%), leaf area (50%), P n (71%), and G s (90%) of winter wheat. Furthermore, the total nonstructural carbohydrates were accumulated 10% and 27% and increased R d by 127% and 99% when subjected to water × warming and e[CO2] × water × warming. These results suggest that water × warming may cause irreversible damage in winter wheat and thus the effect of "CO2 fertilization effect" may be overestimated by the current process-based ecological model.

SIRT1 downregulation provokes immune-inflammatory responses in hair follicle outer root sheath cells and may contribute to development of alopecia areata.

BACKGROUND: Silent information regulator 1 (SIRT1), a type III histone deacetylase, is involved in various cutaneous and systemic autoimmune diseases including systemic lupus erythematosus, rheumatoid arthritis, and psoriasis. However, little is known about the role of SIRT1 in the development of alopecia areata (AA). OBJECTIVES: This study investigated whether SIRT1 regulates the hair follicle immune system and is involved in AA pathogenesis. METHODS: SIRT1 expression in human scalp tissue was analyzed using immunohistochemical staining, qPCR, and western blotting. The regulatory effect of SIRT1 was evaluated after stimulation with the double-stranded RNA mimic polyinosinic:polycytidylic acid (poly I:C) in hair follicle outer root sheath (ORS) cells and C3H/HeJ mice. RESULTS: SIRT1 expression was significantly reduced in the AA scalp compared to the normal scalp. SIRT1 inhibition upregulated MHC class I polypeptide-related sequence A and UL16 binding protein 3 in hair follicle ORS cells. SIRT1 inhibition also promoted the production of Th1 cytokines (IFN-γ and TNF-α), IFN-inducible chemokines (CXCL9 and CXCL10), and T cell migration in ORS cells. Conversely, SIRT1 activation suppressed the autoreactive inflammatory responses. The counteractive effect of the immune response by SIRT1 was mediated through the deacetylation of NF-κB and phosphorylation of STAT3. CONCLUSION: SIRT1 downregulation induces immune-inflammatory responses in hair follicle ORS cells and may contribute to AA development.

The CO2 fertilization effect on leaf photosynthesis of maize (Zea mays L.) depends on growth temperatures with changes in leaf anatomy and soluble sugars.

Understanding the potential mechanisms and processes of leaf photosynthesis in response to elevated CO2 concentration ([CO2]) and temperature is critical for estimating the impacts of climatic change on the growth and yield in crops such as maize (Zea mays L.), which is a widely cultivated C4 crop all over the world. We examined the combined effect of elevated [CO2] and temperature on plant growth, leaf photosynthesis, stomatal traits, and biochemical compositions of maize with six environmental growth chambers controlling two CO2 levels (400 and 800 µmol mol-1) and three temperature regimes (25/19°C, 31/25°C, and 37/31°C). We found that leaf photosynthesis was significantly enhanced by increasing growth temperature from 25/19°C to 31/25°C independent of [CO2]. However, leaf photosynthesis drastically declined when the growth temperature was continually increased to 37/31°C at both ambient CO2 concentration (400 µmol mol-1, a[CO2]) and elevated CO2 concentration (800 µmol mol-1, e[CO2]). Meanwhile, we also found strong CO2 fertilization effect on maize plants grown at the highest temperature (37/31°C), as evidenced by the higher leaf photosynthesis at e[CO2] than that at a[CO2], although leaf photosynthesis was similar between a[CO2] and e[CO2] under the other two temperature regimes of 25/19°C and 31/25°C. Furthermore, we also found that e[CO2] resulted in an increase in leaf soluble sugar, which was positively related with leaf photosynthesis under the high temperature regime of 37/31°C (R 2 = 0.77). In addition, our results showed that e[CO2] substantially decreased leaf transpiration rates of maize plants, which might be partially attributed to the reduced stomatal openness as demonstrated by the declined stomatal width and stomatal area. These results suggest that the CO2 fertilization effect on plant growth and leaf photosynthesis of maize depends on growth temperatures through changing stomatal traits, leaf anatomy, and soluble sugar contents.

A specific reverse complement sequence for distinguishing Brucella canis from other Brucella species.

Canine brucellosis is primarily caused by Brucella canis, but other Brucella species can also cause the disease. Identifying sequences specific to B. canis and establishing PCR assays that can distinguish between B. canis and other Brucella species is essential to determine the etiology of canine brucellosis and the source of infection and to achieve effective control. We analyzed the gaps and SNPs of genomes I and II from B. canis strain RM6/66 and B. melitensis strain 16M using the Mauve genome alignment software, and the specificity of each of these differential regions was analyzed by BLAST. A 132 bp specific sequence was found between the DK60_915 (glycosyl hydrolase 108 family protein) and DK60_917 (aldose 1-epimerase) loci in B. canis chromosome 1. Further comparative analysis revealed that this is a reverse complement sequence between B. canis and other Brucella species. Then, three primers were designed based on the sequence that could detect B. canis with a 310 bp amplification product or other Brucella species with a 413 bp product. The PCR based on these primers had reasonable specificity and a sensitivity of 100 copies of Brucella DNA. The detection results for the blood samples of the aborted dogs showed a favorable accordance with the Bruce-ladder multiplex PCR assay. In conclusion, we found a specific reverse complement sequence between B. canis and other Brucella and developed a PCR method that allows a more comprehensive identification of the pathogen involved in canine brucellosis. These findings provide an effective means for preventing and controlling brucellosis.

Repurposing the anthelmintic praziquantel to treat psoriasis.

BACKGROUND AND PURPOSE: The anthelmintic drug praziquantel has been used as a standard treatment for schistosomiasis for over 40 years. This study aimed to repurpose praziquantel to treat psoriasis. EXPERIMENTAL APPROACH: Psoriasis-like skin inflammation was induced in mice (C57 and Balb/C) by topical application of imiquimod or intradermal injection of recombinant IL-23. Praziquantel was either orally or topically administered during the psoriasis induction period. KEY RESULTS: Mice treated with either oral or topical praziquantel exhibited markedly improved psoriasiform skin symptoms when compared with control mice, as judged by disease severity score, epidermal thickening, inflammatory cell infiltration and spleen size. Flow cytometric analysis of infiltrating immune cells from mouse skin displayed reduced infiltration of Th17 cells. In vitro experiments revealed that praziquantel inhibited STAT3 phosphorylation and RORγt expression in splenic CD4+ T-cells. Praziquantel also decreased STAT3 phosphorylation in HEK-A/F cells. Down-regulation of STAT3 phosphorylation in these cells accounts for the decreased number of Th17 cells and keratinocytes. CONCLUSION AND IMPLICATIONS: These results provide the first preclinical evidence that praziquantel may effectively treat psoriasis, and suggest that praziquantel alleviates symptoms in mice by inhibiting STAT3 phosphorylation, thereby suppressing Th17 immune responses.

2'-Hydroxycinnamaldehyde ameliorates imiquimod-induced psoriasiform inflammation by targeting PKM2-STAT3 signaling in mice.

2'-Hydroxycinnamaldehyde (HCA), the active component isolated from the stem bark of Cinnamomum cassia, exerts anticancer effects through multiple mechanisms. We recently determined that HCA inhibits signal transducer and activator of transcription 3 (STAT3) signaling in prostate cancer cells. Because STAT3 overactivation has been closely associated with the development of psoriasis, a chronic autoimmune skin disease, we examined whether HCA ameliorates skin lesions in an imiquimod-induced psoriasis-like mouse model. The results showed that intraperitoneal administration of HCA alleviated imiquimod-induced psoriasis-like dermatitis, epidermal thickening, dermal infiltration of inflammatory cells, and proinflammatory cytokine production. Mechanistically, HCA inhibited pyruvate kinase isozyme M2 and STAT3 signaling, leading to the suppression of T cell activation, Th17 cell differentiation, and keratinocyte hyperproliferation. These results suggest that HCA may be a new treatment for psoriasis and other STAT3-mediated skin disorders, such as infection, inflammation and carcinogenesis.

Inhibiting Protein Kinase Activity of Pyruvate Kinase M2 by SIRT2 Deacetylase Attenuates Psoriasis.

Signal transducer and activator of transcription 3 (STAT3) is crucial for the pathogenesis of psoriasis. Studies describe pleiotropic roles for a glycolytic enzyme pyruvate kinase M2 (PKM2) as a nuclear kinase of STAT3. However, little is known about the function of PKM2 in T helper type 17 cells in association with STAT3. In this study, we investigated whether and how SIRT2 deacetylase regulated the protein kinase function of PKM2 in T helper type 17 cell‒mediated inflammatory responses in psoriasis. Sirt2 knockout mice and wild-type littermates had psoriatic dermatitis induced by topical treatment of imiquimod or intradermal injection of recombinant IL-23. An initial downregulation of SIRT2 and an increase in PKM2 acetylation and STAT3 phosphorylation were observed in psoriasiform lesions of mice. SIRT2 directly interacted with and deacetylated PKM2 to suppress STAT3 phosphorylation. Consequently, psoriasiform skin inflammation was aggravated in Sirt2 knockout mice. Conversely, genetic re-expression of Sirt2 or pharmacological blockade of PKM2 decreased the disease severity. Flow cytometric analysis of skin tissues of Sirt2 knockout mice showed enhanced infiltration of T helper type 17 cells. Ex vivo experiments showed that SIRT2 deficiency accelerated T helper type 17 cell differentiation with the concomitant production of IL-17A and IL-22. The results suggest SIRT2-mediated PKM2 deacetylation as an effective option for psoriasis therapy.

p21-activated kinase 4 phosphorylates peroxisome proliferator-activated receptor Υ and suppresses skeletal muscle regeneration.

BACKGROUND: Skeletal muscle regeneration is an adaptive response to injury that is crucial to the maintenance of muscle mass and function. A p21-activated kinase 4 (PAK4) serine/threonine kinase is critical to the regulation of cytoskeletal changes, cell proliferation, and growth. However, PAK4's role in myoblast differentiation and regenerative myogenesis remains to be determined. METHODS: We used a mouse model of myotoxin (notexin)-induced muscle regeneration. In vitro myogenesis was performed in the C2C12 myoblast cell line, primary myoblasts, and primary satellite cells. In vivo overexpression of PAK4 or kinase-inactive mutant PAK4S474A was conducted in skeletal muscle to examine PAK4's kinase-dependent effect on muscle regeneration. The regeneration process was evaluated by determining the number and size of multinucleated myofibres and expression patterns of myogenin and eMyHC. To explore whether PAK4 inhibition improves muscle regeneration, mice were injected intramuscularly with siRNA that targeted PAK4 or orally administered with a chemical inhibitor of PAK4. RESULTS: p21-activated kinase 4 was highly expressed during the myoblast stage, but expression gradually and substantially decreased as myoblasts differentiated into myotubes. PAK4 overexpression, but not kinase-inactive mutant PAK4S474A overexpression, significantly impeded myoblast fusion and MyHC-positive myotube formation in C2C12 cells, primary myoblasts, and satellite cells (P < 0.01). Conversely, PAK4 silencing led to an 8.7% and a 20.3% increase in the number of multinucleated larger myotubes in C2C12 cells and primary myoblasts. Further, in vivo overexpression of PAK4 by adenovirus injection to mice prior to and after myotoxin-induced injury led to a 52.6% decrease in the number of eMyHC-positive myofibres on Day 5 in tibialis anterior muscles as compared with those injected with control adenoviruses (P < 0.01), while Ad-PAK4S474A showed comparable muscle regeneration parameters. PAK4-induced repression of muscle regeneration coincided with an increase in phosphatase and tensin homologue (PTEN) expression and a decrease in phosphoinositide 3-kinase-Akt signalling. In contrast, PAK4 silencing reduced PTEN expression in mice. Consistent with these findings, prodrug of PAK4 inhibitor CZh-226 (30 mg/kg) orally administered to mice repressed PTEN expression and accelerated myotube formation. Subsequent mechanistic studies revealed that PAK4 directly phosphorylates PPARγ at S273 to increase its transcription activity, thereby up-regulating PTEN expression. Importantly, an analysis of the Genotype-Tissue Expression database showed a positive correlation between PAK4 and PTEN in human skeletal muscle tissues (P < 0.01). CONCLUSIONS: p1-activated kinase 4 is a new member of PPARγ kinase, and PAK4 inhibition may have a therapeutic role as an accelerant of muscle regeneration.

Histone deacetylase 8 inhibition alleviates cholestatic liver injury and fibrosis.

Cholestasis is a pathological condition involving blockage of bile flow that results in hepatotoxicity, inflammation, and fibrosis. Although recent studies have shown that histone deacetylases (HDACs) are involved in the progression of fibrosis in various organs, the role of HDAC8 on liver fibrosis has until now remained unexplored. This study presents a newly-synthesized, selective HDAC8 inhibitor SPA3014 composed of a vinyl disulfide-sulfoxide core, and evaluates its therapeutic efficacy against cholestatic liver injury and fibrosis in bile duct-ligated (BDL) mice. We first observed the increase in HDAC8 protein levels in mice with BDL and patients with cholestatic liver disease. Mice with BDL that were pretreated with SPA3014 had lower liver damage and fibrosis, based on gross examination, histopathologic findings, and biochemical analyses, than did vehicle-treated mice. Studies with LX-2 human hepatic stellate cells showed that SPA3014 exerted protective effects by inhibiting TGF-ß-mediated activation of MAPK-Smad2/3 and JAK2-STAT3 pathways and by upregulating PPARγ expression. Overall, these results strongly suggest that HDAC8 inhibition constitutes a new therapeutic strategy for treatment of cholestatic liver injury.

The Mechanism of Extraction of Peanut Protein and Oil Bodies by Enzymatic Hydrolysis of the Cell Wall.

Degradation of the peanut cell wall is a critical step in the aqueous enzymatic extraction process to extract proteins and oil bodies. Viscozyme® L, a compound cell wall degrading enzyme, has been applied as an alternative to protease in the process of aqueous enzymatic extraction, but the mechanism of cell wall enzymolysis remains unclear. The present study aims to investigate the changes in cellulose, hemicellulose, and pectin content of the peanut cell wall hydrolyzed by Viscozyme® L. The degree to which the main components of the peanut cell wall, such as trans-1, 2-cyclohexanediamine-N,N,N',N'-acetic acid-soluble pectin (CDTA-soluble pectin), Na2CO3-soluble pectin, cellulose, and hemicellulose, are degraded is closely related to the extraction of oil bodies and peanut protein at different solid-liquid ratio of powered peanut seed in distilled water, enzyme concentration, enzyme hydrolysis temperature, and enzyme hydrolysis time. The key sites of Viscozyme® L activity on cell wall polysaccharides were explored by comparing the changes in chemical bonds under different extraction conditions using Fourier-transform infrared spectroscopy (FT-IR) absorption bands and principal component analysis (PCA). Viscozyme® L acted on the C-O stretching, C-C stretching, and CH2 symmetrical bending of cellulose, the C-O stretching and O-C-O asymmetrical bending of hemicellulose, and the C-O stretching and C-C stretching of pectin.

ERRγ suppression by Sirt6 alleviates cholestatic liver injury and fibrosis.

Orphan nuclear receptor estrogen-related receptor γ (ERRγ) stimulates bile acid production; however, the role and the regulatory mechanism of ERRγ in cholestatic liver disease are largely unknown. This study identifies that Sirt6 is a deacetylase of ERRγ and suggests a potentially novel mechanism by which Sirt6 activation alleviates cholestatic liver damage and fibrosis through regulating ERRγ. We observed that hepatic expression of Sirt6 is repressed, whereas hepatic expression of ERRγ is upregulated in murine cholestasis models. Hepatocyte-specific Sirt6-KO mice were more severely injured after a bile duct ligation (BDL) than WT mice, and adenoviral reexpression of Sirt6 reversed liver damage and fibrosis as demonstrated by biochemical and histological analyses. Mechanistically, Sirt6 deacetylated ERRγ, thereby destabilizing ERRγ and inhibiting its transcriptional activity. Elimination of hepatic ERRγ using Ad-shERRγ abolished the deleterious effects of Sirt6 deficiency, whereas ERRγ overexpression aggravated cholestatic liver injury. Administration of a Sirt6 deacetylase activator prevented BDL-induced liver damage and fibrosis. In patients with cholestasis, Sirt6 expression was decreased, whereas total ERRγ and acetylated ERRγ levels were increased, confirming negative regulation of ERRγ by Sirt6. Thus, Sirt6 activation represents a potentially novel therapeutic strategy for treating cholestatic liver injury.