Comprehensive single-cell analysis deciphered microenvironmental dynamics and immune regulator olfactomedin 4 in pathogenesis of gallbladder cancer.

OBJECTIVE: Elucidating complex ecosystems and molecular features of gallbladder cancer (GBC) and benign gallbladder diseases is pivotal to proactive cancer prevention and optimal therapeutic intervention. DESIGN: We performed single-cell transcriptome analysis on 230 737 cells from 15 GBCs, 4 cholecystitis samples, 3 gallbladder polyps, 5 gallbladder adenomas and 16 adjacent normal tissues. Findings were validated through large-scale histological assays, digital spatial profiler multiplexed immunofluorescence (GeoMx), etc. Further molecular mechanism was demonstrated with in vitro and in vivo studies. RESULTS: The cell atlas unveiled an altered immune landscape across different pathological states of gallbladder diseases. GBC featured a more suppressive immune microenvironment with distinct T-cell proliferation patterns and macrophage attributions in different GBC subtypes. Notably, mutual exclusivity between stromal and immune cells was identified and remarkable stromal ecosystem (SC) heterogeneity during GBC progression was unveiled. Specifically, SC1 demonstrated active interaction between Fibro-iCAF and Endo-Tip cells, correlating with poor prognosis. Moreover, epithelium genetic variations within adenocarcinoma (AC) indicated an evolutionary similarity between adenoma and AC. Importantly, our study identified elevated olfactomedin 4 (OLFM4) in epithelial cells as a central player in GBC progression. OLFM4 was related to T-cell malfunction and tumour-associated macrophage infiltration, leading to a worse prognosis in GBC. Further investigations revealed that OLFM4 upregulated programmed death-ligand 1 (PD-L1) expression through the MAPK-AP1 axis, facilitating tumour cell immune evasion. CONCLUSION: These findings offer a valuable resource for understanding the pathogenesis of gallbladder diseases and indicate OLFM4 as a potential biomarker and therapeutic target for GBC.

Non-muscle myosin II knockdown improves survival and therapeutic effects of implanted bone marrow-derived mesenchymal stem cells in lipopolysaccharide-induced acute lung injury.

BACKGROUND: Bone marrow-derived mesenchymal stem cells (BMSCs) have been shown to have some beneficial effects in acute lung injury (ALI), but the therapeutic effects are limited due to apoptosis or necrosis after transplantation into injured lungs. Here, we aim to explore whether Non-muscle myosin II (NM-II) knockdown could enhance BMSCs survival and improve therapeutic effects in ALI. METHODS: MSCs, isolated from rat bone marrow, were transfected with the small interfering RNA (siRNA) targeted to NM-II mRNA by a lentivirus vector. Rats were equally randomized to four groups: the control group was given normal saline via tail vein; the other three groups underwent intratracheal lipopolysaccharide (LPS) instillation followed by administration with either normal saline, BMSCs transduced with lentivirus-enhanced green fluorescent protein (eGFP) empty vector, or BMSCs transduced with lentivirus-eGFP NM-II siRNA. Hematoxylin and eosin staining was used to evaluate lung histopathologic changes and Masson trichrome staining was used to assess lung fibrosis. The myeloperoxidase activity was also tested in lung tissues. The mRNA expression of inflammatory cytokines in lung tissues was determined via quantitative reverse transcription PCR. Sex-determining region of the Y chromosome gene expression was measured by fluorescence in situ hybridization (FISH) assay. The expression of self-renewal activity and apoptosis-associated proteins were measured by Western blot. RESULTS: Transplantation of NM-II siRNA-modified BMSCs could improve histopathological morphology, decrease inflammatory infiltrates, down-regulate the expression levels of inflammatory cytokines, and reduce pulmonary interstitial edema. NM-II siRNA-modified BMSCs showed antifibrotic properties and alleviated the degrees of pulmonary fibrosis induced by endotoxin. In addition, NM-II knockdown BMSCs showed slightly better therapeutic effect on lung inflammation when compared with control BMSCs. The beneficial effects of NM-II siRNA-modified BMSCs may be attributed to enhanced self-renewal activity and decreased apoptosis. CONCLUSIONS: NM-II knockdown could inhibit the apoptosis of implanted BMSCs in lung tissues and improve its self-renewal activity. NM-II siRNA-modified BMSCs have a slightly enhanced ability to attenuate lung injury after LPS challenge.

A review on mushroom-derived bioactive peptides: Preparation and biological activities.

Mushroom bioactive peptides (MBAPs) refer to bioactive peptides extracted directly or indirectly from mushrooms or their mycelia. Owing to the presence of a large quantity of high-quality proteins, many mushrooms are promising sources of bioactive peptides. The beneficial effects, including antihypertensive, antioxidant, and antimicrobial activities, of MBAPs are being increasingly recognized with regards to health promotion and disease prevention. However, this field is relatively undeveloped and relevant reviews are scarce. Hence, the aim of this review was to present the current research status of MBAPs, focusing on their preparation and biological functions. An insight regarding the direction of future research has been also discussed.

Can systemic inflammatory response syndrome score at admission predict clinical outcome in patients with severe burns?

INTRODUCTION: The use of SIRS score as a predictor of outcomes in patients with severe burns has not been fully evaluated. Here, we aimed to test that whether admission SIRS score, combining with other predictors, could be used in predicting outcomes in patients with severe burns. Additionally, we compared the prognostic accuracy of admission SIRS score with other score systems and newly developed models. METHODS: We performed a retrospective study of adult patients with ≥40% total body surface area burns admitted to a burn center from 2005 to 2017. The primary outcome was in-hospital mortality, and the secondary outcomes were hospital and intensive care unit length of stay. SIRS score, rBaux score, ABSI and newly developed models were compared using area under the receiver operating characteristic curve analysis. RESULTS: Out of the total 144 patients, 128 (88.9%) met SIRS criteria on admission; with a predominant SIRS scores of 3. Patients with admission SIRS were more likely to have larger burns and have higher rBaux and ABSI scores when compared with non-SIRS patients. With the each increment of admission SIRS score, total and full-thickness burn areas, proportion of inhalation injury, tracheostomy and mortality increased significantly. However, SIRS score at admission was not increasingly predictive of deleterious outcomes when analyzed by multivariable regression analysis. Although the combination of SIRS score, age, and burn-specific variables showed better or equal prognostication of outcomes than that of other score systems, the contribution of the variable SIRS score was negligible. CONCLUSIONS: The model with the variables age, percentage full-thickness burns, and inhalation injury provided excellent prediction of poor outcomes in patients with severe burns, while SIRS score has limited use for prognostic determinations.

Blood transfusions in severe burn patients: Epidemiology and predictive factors.

BACKGROUND: Blood is a vital resource commonly used in burn patients; however, description of blood transfusions in severe burns is limited. The purpose of this study was to describe the epidemiology of blood transfusions and determine factors associated with increased transfusion quantity. METHODS: This is a retrospective study of total 133 patients with >40% total body surface area (TBSA) burns admitted to the burn center of Changhai hospital from January 2008 to December 2013. The study characterized blood transfusions in severe burn patients. Univariate and Multivariate regression analyses were used to evaluate the association of clinical variables with blood transfusions. RESULTS: The overall transfusion rate was 97.7% (130 of 133). The median amount of total blood (RBC and plasma), RBC and plasma transfusions was 54 units (Interquartile range (IQR), 20-84), 19 units (IQR, 4-37.8) and 28.5 units (IQR, 14.8-51.8), respectively. The number of RBC transfusion in and outside operation room was 7 (0, 14) and 11 (2, 20) units, and the number of plasma was 6 (0.5, 12) and 21 (11.5, 39.3) units. A median of one unit of blood was transfused per TBSA and an average of 4 units per operation was given in the series. The consumption of plasma is higher than that of RBC. On multivariate regression analysis, age, full-thickness TBSA and number of operations were significant independent predictors associated with the number of RBC transfusion, and coagulopathy and ICU length showed a trend toward RBC consumption. Predictors for increased plasma transfusion were female, high full-thickness TBSA burn and more operations. CONCLUSIONS: Severe burn patients received an ample volume of blood transfusions. Fully understanding of predictors of blood transfusions will allow physicians to better optimize burn patients during hospitalization in an effort to use blood appropriately.

Sequence, taste and umami-enhancing effect of the peptides separated from soy sauce.

Five tasty peptides were separated from soy sauce, by sensory-guided fractionation, using macroporous resin, medium-pressure liquid chromatography and reverse phase-high performance liquid chromatography, and identified by ultra-performance liquid chromatography tandem mass-spectrometry as ALPEEV, LPEEV, AQALQAQA, EQQQQ and EAGIQ (which originated from glycinin A1bB2-445, glycinin A1bB2-445, cobyric acid synthase, leucine-tRNA ligase and glycoprotein glucosyltransferase, respectively). LPEEV, AQALQAQA and EQQQQ tasted umami with threshold values of 0.43, 1.25 and 0.76mmol/l, respectively. ALPEEV and EAGIQ had minimal umami taste, but ALPEEV, EAGIQ and LPEEV showed umami-enhancement with a threshold estimated at 1.52, 1.94 and 3.41mmol/l, respectively. In addition, the synthetic peptides showed much better sensory taste than mixtures of their constitutive amino acids. It indicated that peptides might play an important role in the umami taste of soy sauce.

Macroporous resin purification of peptides with umami taste from soy sauce.

In this study, the performance and separation characteristics of four macroporous resins for purifying umami peptides from soy sauce were examined. Results showed that the resins could separate the peptides of soy sauce, and the particle diffusion kinetics model was suitable for describing the whole exothermic (ΔH < 43 kJ/mol) adsorption process on the SP-825 and HP-20 resins, while the pseudo-second-order kinetics model accurately described the XAD-16 and HP-2 MGL resins. Furthermore, the adsorption processes of the peptides followed the Freundlich model. The XAD-16 resin was the most effective resin for the enrichment of peptides due to its high adsorption and total desorption capacities. Interestingly, the umami peptides were enriched in the deionized water fraction. This study provides new insights into exploring performance and separation characteristics of macroporous resins on soy sauce, and indicated that peptide may be the contributor to the umami taste in Chinese soy sauce.

Identification of antioxidative peptides from defatted walnut meal hydrolysate with potential for improving learning and memory.

Defatted walnut meal (DWM), a main byproduct of walnut oil production, is rich in proteins with a high essential amino acid content. Our research was focused on the functional activities of defatted walnut meal hydrolysate (DWMH). DWMH exhibited relatively strong hydroxyl scavenging and oxygen radical absorbance capacities, protective effect on H2O2-injured PC12 cells compared with GSH and cerebrolysin. Besides, DWMH could combat d-galactose induced learning and memory impairments in the Morris water maze test and in the Dark/light avoidance test of mice. Seventy-seven peptides were identified by UPLC-ESI-MS/MS in the most potent antioxidative fraction of DWMH. WSREEQEREE and ADIYTEEAGR were the peptides that most likely accounted for the suppression of H2O2-induced apoptosis in PC12 cells. The mixture of the commercially available amino acids did not offer any protection on H2O2-injured PC12 cells, suggesting that amino acid sequence rather than amino acid composition was the dominant influencing factor. DWMH could be recommended as natural antioxidants for the development of functional foods targeting memory impairments.

Absorption and desorption behaviour of the flavonoids from Glycyrrhiza glabra L. leaf on macroporous adsorption resins.

The kinetics of adsorption and desorption behaviours of five macroporous resins for enriching flavonoids from Glycyrrhizaglabra L. leaf were investigated. All five resins showed similar and effective adsorption and desorption properties. A pseudo-second-order kinetics model was suitable for evaluating the whole adsorption process. Additionally, two representative resins (XAD-16 and SP825) were chosen for adsorption thermodynamics study. The adsorption of the representative resins was an exothermic and physical adsorption process. Further column chromatography of XAD-16 and SP825 showed that the total flavonoids (from 16.8% to 55.6% by XAD-16 and to 53.9% by SP825) and pinocembrin (from 5.49% to 15.2% by XAD-16 and to 19.8% by SP825) were enriched in 90% ethanol fractions. Meanwhile, the antioxidant capacities and nitrite-scavenging capacities were 2-3times higher than those of the crude extract. The fractions with high flavonoid and pinocembrin contents could be used as biologically active ingredients in functional food.

Effusanin E suppresses nasopharyngeal carcinoma cell growth by inhibiting NF-κB and COX-2 signaling.

Rabdosia serra is well known for its antibacterial, anti-inflammatory and antitumor activities, but no information has been available for the active compounds derived from this plant in inhibiting human nasopharyngeal carcinoma (NPC) cell growth. In this study, we isolated and purified a natural diterpenoid from Rabdosia serra and identified its chemical structure as effusanin E and elucidated its underlying mechanism of action in inhibiting NPC cell growth. Effusanin E significantly inhibited cell proliferation and induced apoptosis in NPC cells. Effusanin E also induced the cleavage of PARP, caspase-3 and -9 proteins and inhibited the nuclear translocation of p65 NF-κB proteins. Moreover, effusanin E abrogated the binding of NF-κB to the COX-2 promoter, thereby inhibiting the expression and promoter activity of COX-2. Pretreatment with a COX-2 or NF-κB-selective inhibitor (celecoxib or ammonium pyrrolidinedithiocarbamate) had an additive effect on the effusanin E-mediated inhibition of proliferation, while pretreatment with an activator of NF-κB/COX-2 (lipopolysaccharides) abrogated the effusanin E-mediated inhibition of proliferation. Effusanin E also significantly suppressed tumor growth in a xenograft mouse model without obvious toxicity, furthermore, the expression of p50 NF-κB and COX-2 were down-regulated in the tumors of nude mice. These data suggest that effusanin E suppresses p50/p65 proteins to down-regulate COX-2 expression, thereby inhibiting NPC cell growth. Our findings provide new insights into exploring effusanin E as a potential therapeutic compound for the treatment of human nasopharyngeal carcinoma.

Bioactive profiles, antioxidant activities, nitrite scavenging capacities and protective effects on H2O2-injured PC12 cells of Glycyrrhiza glabra L. leaf and root extracts.

This study compared the total flavonoid content of Glycyrrhiza glabra L. leaf and root extracts. Results suggested that the total flavonoid content in the leaf extract was obviously higher than that in the root extract. Pinocembrin, the main compound in the leaf extract after purification by column chromatography, showed good antioxidant activity and nitrite scavenging capacity, but moderate inhibitory effect on mushroom tyrosinase. Liquiritin was the main compound in root extract and possessed strong inhibitory effect on mushroom tyrosinase. Both compounds exhibited significant protection effect on H2O2-injured PC12 cells at a low concentration. These results indicate that Glycyrrhiza glabra L. leaf is potential as an important raw material for functional food.

GC/MS analysis of volatiles obtained by headspace solid-phase microextraction and simultaneous-distillation extraction from Rabdosia serra (MAXIM.) HARA leaf and stem.

Volatiles in Rabdosia serra were investigated by headspace solid phase microextraction (HS-SPME) and simultaneous-distillation extraction (SDE). The HS-SPME technique was previously evaluated to optimise sampling conditions. A total of 56 and 48 compounds including alcohols, aldehydes, hydrocarbons, ketones, carboxylic acid, ester, and aromatics were identified in leaf and stem by optimised HS-SPME method (CAR/PDMS fibre; incubation time, 10 min; extraction temperature, 50°C; extraction time, 40 min), respectively. 1-Octen-3-ol and (2E)-hexenal had significant contribution to R. serra aroma. Cluster analysis indicated that leaf and stem exhibited different volatile diversity. Air drying was favourable for the retention of the volatiles, while freeze- and sun-drying led to the loss of volatiles. SDE method preferred to the analysis of compounds with low volatility including fatty acids and esters. HS-SPME was a useful technique for the analysis of readily volatile components for the characteristics of R. serra aroma.

Structural characteristics of water-soluble polysaccharides from Rabdosia serra (MAXIM.) HARA leaf and stem and their antioxidant capacities.

Water-soluble polysaccharides of Rabdosia serra leaf and stem were fractionated by ultrafiltration and DEAE-Sepharose fast flow chromatogram to obtain water (RSLP-I and RSSP-I), 0.1M NaCl (RSLP-II and RSSP-II) and 0.2M NaCl (RSLP-III and RSSP-III) eluates. Their molecular weights were determined by high performance gel permeation chromatography. Monosaccharide composition analysis indicated that the water eluates comprised of rhamnose, arabinose, xylose, mannose, glucose and galactose. The ascending percentage of galactose and descending percentage of glucose in the eluates were observed with the increase of NaCl concentration. The branched RSLP-I and RSSP-I were composed mainly of →6)-Glcp-(1→, →6)-Galp-(1→, and →5)-Araf-(1→ residues with the ratio of 15.0:4.2:3.8 and 5.5:6.4:3.5, respectively. However, a low level of 1,6-linked glucosyl was observed in RSLP-II, RSSP-II, RSLP-III and RSSP-III. The structural characteristics were further analysed by infrared spectrophotometry. The purified leaf and stem polysaccharides possessed moderate antioxidant capacities.