Analysis of Volatile Aroma Components in Different Parts of Shiitake Mushroom (Lentinus edodes) Treated with Ultraviolet C Light-Emitting Diodes Based on Gas Chromatography-Ion Mobility Spectroscopy.

Further assessment of ultraviolet C light-emitting diode (UVC-LED) irradiation for influencing shiitake mushrooms' (Lentinus edodes) volatile and sensory properties is needed. In this study, a comparison of UVC-LED irradiation treatment on the flavor profiles in various parts of shiitake mushrooms was conducted using gas chromatography-ion mobility spectrometry (GC-IMS) and sensory analysis. Sixty-three volatile compounds were identified in shiitake mushrooms. The fresh shiitake mushrooms were characterized by the highest values of raw mushroom odors. After UVC-LED treatment, the content of C8 alcohols decreased, especially that of 1-octen-3-ol, while the content of aldehydes increased, especially the content of nonanal and decanal. The score of fatty and green odors was enhanced. For fresh samples, the mushroom odors decreased and the mushroom-like odors weakened more sharply when treated in ethanol suspension than when treated with direct irradiation. The fruit odors were enhanced using direct UVC-LED irradiation for fresh mushroom samples and the onion flavor decreased. As for shiitake mushroom powder in ethanol suspension treated with UVC-LED, the sweaty and almond odor scores decreased and the vitamin D2 content in mushroom caps and stems reached 668.79 µg/g (dw) and 399.45 µg/g (dw), respectively. The results obtained from this study demonstrate that UVC-LED treatment produced rich-flavored, quality mushroom products.

Successful Ultrasound-Guided Superficial Cervical Plexus Block Treatment for Head and Neck Pain with an Unusually Delayed Onset Following Ventriculoperitoneal Shunt: A Case Report.

Background and Objectives: Ventriculoperitoneal (VP) shunt placement is the most common treatment for cerebrospinal fluid diversion. Head and neck pain occurring after a long period following VP shunt insertion is rarely reported. Here, we present a rare case of head and neck pain occurring 2 years after surgery due to irritation of the superficial cervical plexus by the VP shunt. Case Description: A 46-year-old female patient received VP shunt placement surgery. Two years after the surgery, she experienced a left temporal headache with neck pain on the left side, which extended to the left para-auricular and fascial region. Ultrasound (US) scanning revealed that the VP shunt passed within the superficial cervical fascia and through the left sternocleidomastoid muscle (SCM). Additionally, friction of the branches of the superficial cervical plexus and of the greater auricular and lesser occipital nerves caused by the VP shunt was found underneath the lateral border of the SCM. Subsequently, the blocking and hydro-release of the left superficial cervical plexus were performed. After four series of treatments, the patient's head and neck pain vanished, and the frequency of the headaches was substantially reduced. The patient was regularly followed-up in the outpatient department of neurosurgery. Conclusions: Head and neck pain caused by the malpositioning of a VP shunt catheter with an unusually delayed onset is a rarely reported complication and could be easily neglected. Patients with head and neck pain following VP shunt insertion should be checked using US scanning to identify the potential origin of the pain and receive adequate treatments. Intraoperative US-guided tunnelling is suggested to avoid the malpositioning of the VP shunt catheter.

Associations of parecoxib and other variables with recovery and safety outcomes in total knee arthroplasty: insights from a retrospective cohort study.

Background: Early mobilization post-total knee arthroplasty (TKA) significantly affects patient outcomes. While parecoxib is known to reduce postoperative pain and morphine use with a favorable safety profile, its impact on mobilization timing post-TKA remains uncertain. This retrospective study aims to assess parecoxib's influence on postoperative mobilization timing in TKA patients without compromising safety. Methods: This study included unilateral TKA patients treated for primary knee osteoarthritis under general anesthesia. We divided the study period into two intervals, 2007-2012 and 2013-2018, to evaluate temporal differences. Both the control group and parecoxib group received standard postoperative oral analgesics and as-needed intramuscular morphine. The control group did not receive parecoxib, while the parecoxib group did. Primary outcomes compared postoperative complications and mobilization timing between groups, with secondary outcomes including length of hospital stay (LOS), Visual Analog Scale (VAS) scores for pain, as-needed morphine use, and postoperative nausea/vomiting. Results: Parecoxib did not increase postoperative complications. Unmatched comparison with patients in controlled group found that patients in parecoxib group had significantly shortened mobilization time (2.2 ± 1.1 vs. 2.7 ± 1.6 days, P < 0.001) and LOS (6.7 ± 2.5 vs. 7.2 ± 2.1 days, P = 0.01). Multivariate analysis linked parecoxib use with faster mobilization (ß = -0.365, P < 0.001) but not LOS. Males showed increased mobilization time and LOS compared to females during the period of 2007-2018, but gender had no significant association with LOS during the period of 2013-2018. The 2013-2018 period saw significant reductions in both mobilization time and LOS. Use of a tourniquet and local infiltration analgesia showed no significant impact. ASA classification 1-2 was positively associated with faster mobilization but not LOS. Longer operation times were linked to delayed mobilization and increased LOS. Conclusion: In this study, intravenous parecoxib injection, female gender, and shorter OP time had consistent positive association with shorter time to mobilization after individual multivariate analysis in 2 different period. The use of parecoxib had consistent no significant association with LOS. Only shorter OP time was consistent positive associated with shorter LOS.

The Use of the Shikani Video-Assisted Intubating Stylet Technique in Patients with Restricted Neck Mobility.

Among all the proposed predictors of difficult intubation defined by the intubation difficulty scale, head and neck movement (motility) stands out and plays as a crucial factor in determining the success rate and the degree of ease on endotracheal intubation. Aside from other airway tools (e.g., supraglottic airway devices), optical devices have been developed and applied for more than two decades and have shown their superiority to conventional direct laryngoscopes in many clinical scenarios and settings. Although awake/asleep flexible fiberoptic bronchoscopy is still the gold standard in patients with unstable cervical spines immobilized with a rigid cervical collar or a halo neck brace, videolaryngoscopy has been repeatedly demonstrated to be advantageous. In this brief report, for the first time, we present our clinical experience on the routine use of the Shikani video-assisted intubating stylet technique in patients with traumatic cervical spine injuries immobilized with a cervical stabilizer and in a patient with a stereotactic headframe for neurosurgery. Some trouble-shooting strategies for this technique are discussed. This paper demonstrates that the video-assisted intubating stylet technique is an acceptable alternative airway management method in patients with restricted or confined neck motility.

Putative MicroRNA-mRNA Networks Upon Mdfi Overexpression in C2C12 Cell Differentiation and Muscle Fiber Type Transformation.

Mdfi, an inhibitor of myogenic regulatory factors, is involved in myoblast myogenic development and muscle fiber type transformation. However, the regulatory network of Mdfi regulating myoblasts has not been revealed. In this study, we performed microRNAs (miRNAs)-seq on Mdfi overexpression (Mdfi-OE) and wild-type (WT) C2C12 cells to establish the regulatory networks. Comparative analyses of Mdfi-OE vs. WT identified 66 differentially expressed miRNAs (DEMs). Enrichment analysis of the target genes suggested that DEMs may be involved in myoblast differentiation and muscle fiber type transformation through MAPK, Wnt, PI3K-Akt, mTOR, and calcium signaling pathways. miRNA-mRNA interaction networks were suggested along with ten hub miRNAs and five hub genes. We also identified eight hub miRNAs and eleven hub genes in the networks of muscle fiber type transformation. Hub miRNAs mainly play key regulatory roles in muscle fiber type transformation through the PI3K-Akt, MAPK, cAMP, and calcium signaling pathways. Particularly, the three hub miRNAs (miR-335-3p, miR-494-3p, and miR-709) may be involved in both myogenic differentiation and muscle fiber type transformation. These hub miRNAs and genes might serve as candidate biomarkers for the treatment of muscle- and metabolic-related diseases.

Comparison of Ergosterol and Vitamin D2 in Mushrooms Agaricus bisporus and Cordyceps militaris Using Ultraviolet Irradiation Directly on Dry Powder or in Ethanol Suspension.

Vitamin D deficiency is a severe worldwide health issue. Edible mushrooms are an excellent vitamin D2 source and have gained popularity worldwide as a nutritional food. The objective of this study was to investigate the conversion efficiency of ergosterol to vitamin D2 in Agaricus bisporus and Cordyceps militaris mushrooms under ultraviolet (UV) irradiation directly through dry powder or in ethanol suspension (1:20 g/mL, solid to liquid ratio). Several parameters of UV irradiation conditions such as the material form (dry powder or dry powder in ethanol suspension), exposure time (30, 60, or 120 min), wavelength type (UV-C, UV-B, or UV-A), wavelength combination (UV-C plus UV-B, UV-C plus UV-A, UV-B plus UV-A, or UV-C plus UV-B plus UV-A), and wavelength sequence (UV-C → UV-B, UV-C → UV-A, UV-B → UV-A, or UV-C → UV-B → UV-A), were optimized. Under the optimal UV irradiation conditions (dry powder in ethanol suspension irradiated with UV-C at 40 cm for 120 min), vitamin D2 concentrations increased from not detectable to 72 µg/g (dw) in the A. bisporus dry powder and 1104 µg/g (dw) (about 15-fold increase) in the ethanol suspension. After UV irradiation, the vitamin D2 concentration increased from undetectable to 57 µg/g (dw) in the C. militaris dry powder. In contrast, UV irradiation increased the concentration to 877 µg/g (dw) (about 15-fold higher) in the ethanol suspension. Comparison of the effect of various wavelength combinations showed that UV-C irradiation is more effective than UV-A or UV-B. Furthermore, when irradiated by UV-C at a 40 cm irradiation distance in the ethanol suspension, the increase in vitamin D2 in A. bisporus and C. militaris mushrooms was time- or dose-dependent. The conversion rate of vitamin D2 was low to undetectable under dry powder irradiation, but its ergosterol loss rate was higher than in ethanol suspension irradiation. The ergosterol loss rate in dry C. militaris mushrooms was higher than in the dry A. bisporus mushroom powder. Ultraviolet irradiation in ethanol suspension could greatly increase the vitamin D2 concentration than directly on the dry powder and thus make edible mushrooms more practical as a natural vitamin D source for consumers after entirely removing the ethanol.

Rhodiola rosea Rhizome Extract-Mediated Green Synthesis of Silver Nanoparticles and Evaluation of Their Potential Antioxidant and Catalytic Reduction Activities.

The silver nanoparticles (AgNPs) using the rhizome extract of Rhodiola rosea have been reported. However, their antioxidant activity and whether the biogenic AgNPs could be used to catalyze the reduction of hazardous dye or used as fluorescence enhancers are unknown. This study focused on the facile green synthesis of silver nanoparticles using the rhizome aqueous extract of R. rosea (G-AgNPs). We then studied their antioxidant activity and catalytic degradation of hazardous dye Direct Orange 26 (DO26) and Direct Blue 15 (DB15). Their effects on fluorescein's fluorescent properties were also evaluated. The chemical AgNPs (C-AgNPs) were synthesized by reducing solid sodium borohydride (NaBH4), and its above activities were compared with those of G-AgNPs. The formation of G-AgNPs was confirmed by the appearance of brownish-gray color and the surface plasmon resonance (SPR) peak at 437 nm. The biogenic AgNPs were approximately 10 nm in size with a regular spherical shape identified from transmission electron microscopy (TEM) analysis. G-AgNPs exhibited significantly improved 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity than butylated hydroxytoluene (BHT) and C-AgNPs (p < 0.05). The biogenic G-AgNPs were also found to function as an effective green catalyst in reducing DO26 and DB15 by NaBH4, which is superior to C-AgNPs. Furthermore, G-AgNPs showed better fluorescence enhancement activity than C-AgNPs, and the concentration required was lower. When the concentration of the G-AgNP solution was 64 nmol/L, the fluorescence intensity reached the maximum of 5460, with the fluorescence enhancement efficiency of 3.39, and the fluorescence activity was stable within 48 h. This study shows the efficacy of biogenic AgNPs in catalyzing the reduction of hazardous dye DO26 and DB15. Biogenic AgNPs could also be used as fluorescence enhancers in low concentrations.

Mdfi Promotes C2C12 Cell Differentiation and Positively Modulates Fast-to-Slow-Twitch Muscle Fiber Transformation.

Muscle development requires myoblast differentiation and muscle fiber formation. Myod family inhibitor (Mdfi) inhibits myogenic regulatory factors in NIH3T3 cells, but how Mdfi regulates myoblast myogenic development is still unclear. In the present study, we constructed an Mdfi-overexpression (Mdfi-OE) C2C12 cell line by the CRISPR/Cas9 system and performed RNA-seq on Mdfi-OE and wild-type (WT) C2C12 cells. The RNA-seq results showed that the calcium signaling pathway was the most significant. We also established the regulatory networks of Mdfi-OE on C2C12 cell differentiation and muscle fiber type transformation and identified hub genes. Further, both RNA-seq and experimental verification demonstrated that Mdfi promoted C2C12 cell differentiation by upregulating the expression of Myod, Myog, and Myosin. We also found that the positive regulation of Mdfi on fast-to-slow-twitch muscle fiber transformation is mediated by Myod, Camk2b, and its downstream genes, such as Pgc1a, Pdk4, Cs, Cox4, Acadm, Acox1, Cycs, and Atp5a1. In conclusion, our results demonstrated that Mdfi promotes C2C12 cell differentiation and positively modulates fast-to-slow-twitch muscle fiber transformation. These findings further our understanding of the regulatory mechanisms of Mdfi in myogenic development and muscle fiber type transformation. Our results suggest potential therapeutic targets for muscle- and metabolic-related diseases.

Whole genome sequence analysis reveals genetic structure and X-chromosome haplotype structure in indigenous Chinese pigs.

Chinese indigenous pigs exhibit considerable phenotypic diversity, but their population structure and the genetic basis of agriculturally important traits need further exploration. Here, we sequenced the whole genomes of 24 individual pigs representing 22 breeds distributed throughout China. For comparison with European and commercial breeds (one pig per breed), we included seven published pig genomes with our new genomes for analyses. Our results showed that breeds grouped together based on morphological classifications are not necessarily more genetically similar to each other than to breeds from other groups. We found that genetic material from European pigs likely introgressed into five Chinese breeds. We have identified two new subpopulations of domestic pigs that encompass morphology-based criteria in China. The Southern Chinese subpopulation comprises the classical South Chinese Type and part of the Central China Type. In contrast, the Northern Chinese subpopulation comprises the North China Type, the Lower Yangtze River Basin Type, the Southwest Type, the Plateau Type, and the remainder of the Central China Type. Eight haplotypes and two recombination sites were identified within a conserved 40.09 Mb linkage-disequilibrium (LD) block on the X chromosome. Potential candidate genes (LEPR, FANCC, COL1A1, and PCCA) influencing body size were identified. Our findings provide insights into the phylogeny of Chinese indigenous pig breeds and benefit gene mining efforts to improve major economic traits.

Development of a GC-MS/SIM method for the determination of phytosteryl esters.

A gas chromatography-mass spectrometry (GC-MS) method with selected ion monitoring (SIM) was developed and validated for identification and quantitative analysis of three phytosteryl esters, i.e., campesteryl oletate, stigmasteryl oletate and ß-sitosteryl oletate. The method is simple and efficient and achieved good separation of the three phytosteryl esters in 10 min without saponification and liquid-liquid extraction. A calibration curve for the three phytosteryl esters had a correlation coefficient (R2) better than 0.993. Detection limits were 0.42 mg/mL for campesteryl oletate, 0.32 mg/mL for stigmasteryl oletate and 0.80 mg/mL for ß-sitosteryl oletate. The relative standard deviations (RSD) were within 5.47% for precision and stability for three edible oil samples. Recoveries were from 89.85% to 97.65% for each of the phytosteryl esters. These results suggest that the method can be used to identify and quantify the phytosteryl esters in oil samples.

MiR-199b represses porcine muscle satellite cells proliferation by targeting JAG1.

Pig is a useful medical model for humans due to its similarity in size and physiology. Skeletal muscle plays an essential role in body movement. However, the skeletal muscle injuries are common. Skeletal muscle function maintenance largely depends on preserving the regenerative capacity of muscle. Muscle satellite cells proliferation plays an essential role in postnatal muscle growth and regeneration. Therefore, understanding the mechanisms associated with muscle satellite cells proliferation is essential for devising the alternative treatments for muscle injury. Previous studies showed JAG1-Notch1 signaling pathway and miRNAs regulate the skeletal muscle development. JAG1-Notch1 signal pathway regulates the transcription of certain types of miRNAs which further affects target gene expression. However, the specific relationship between JAG1-Notch1 signal pathway and miRNAs during muscle development has not been established. We found overexpression of intracellular domain of the Notch1 protein (N1ICD) in porcine muscle satellite cells (PSCs) decreased miR-199b level. We demonstrated that miR-199b inhibits PSCs proliferation using the overexpression and inhibition of miR-199b experiment. We also found JAG1, the miR-199b target gene, promotes PSCs proliferation through activating the Notch1 signal pathway. Furthermore, we demonstrated miR-199b forms a feedback loop with the JAG1-Notch1 signal pathway to maintain the PSCs niche homeostasis. Our results of miRNAs and genes work collaboratively in regulating PSCs proliferation expand our understanding in PSCs proliferation mechanism. Furthermore, this finding indicates miR-199b is a potential therapeutic target for muscle atrophy.

Browning of Pig White Preadipocytes by Co-Overexpressing Pig PGC-1α and Mice UCP1.

BACKGROUND/AIMS: Brown adipose tissue (BAT) is critical for mammals' survival in the cold environment. BAT-dependent non-shivering thermogenesis is attributed to uncoupling protein 1 (UCP1)'s disengagement of oxidative phosphorylation from ATP synthesis and dissipates energy as heat. Thus individuals with a substantial amount of BAT are better equipped during cold stress and less likely to become obese. Recently, our laboratory has shown pig adipocytes have no UCP1 protein. The inability of newborn piglets to generate heat contributed to its high death rate. Repairing the genetic defect of UCP1 in pig adipocytes has implications in defending against cold for piglets and developing an alternative treatment for human obesity. METHODS: Q-PCR, western blotting (WB) and oxygen consumption measurement were used to enable functional UCP1 protein in preadipocytes. Immunoprecipitation (IP), chromatin immunoprecipitation (CHIP), and dual-luciferase reporter assay system were used to clarify the thermogenesis mechanism of functional UCP1. RESULTS: Only co-overexpressing mice UCP1 and pig PGC-1α increased not only the mitochondrial number but also the uncoupled respiration rate in the transfected pig adipocytes. The functional mice UCP1 increased the pig PGC-1α activity through the AMPK-SIRT1 pathway. The active form PGC-1α interacted with transcription factors Lhx8, Zic1, ERRα, and PPARα to regulate the expression of mitochondrial energy metabolism and adipocytes browning-related genes. CONCLUSION: Our data suggest a model in which pig PGC-1α and mice UCP1 work collaboratively to restore uncoupling respiration in pig preadipocytes. These results have great implications for piglet survival and developing an alternative treatment for human obesity in the future.

MiR-27b Promotes Muscle Development by Inhibiting MDFI Expression.

BACKGROUND/AIMS: Skeletal muscle plays an essential role in the body movement. However, injuries to the skeletal muscle are common. Lifelong maintenance of skeletal muscle function largely depends on preserving the regenerative capacity of muscle. Muscle satellite cells proliferation, differentiation, and myoblast fusion play an important role in muscle regeneration after injury. Therefore, understanding of the mechanisms associated with muscle development during muscle regeneration is essential for devising the alternative treatments for muscle injury in the future. METHODS: Edu staining, qRT-PCR and western blot were used to evaluate the miR-27b effects on pig muscle satellite cells (PSCs) proliferation and differentiation in vitro. Then, we used bioinformatics analysis and dual-luciferase reporter assay to predict and confirm the miR-27b target gene. Finally, we elucidate the target gene function on muscle development in vitro and in vivo through Edu staining, qRT-PCR, western blot, H&E staining and morphological observation. RESULT: miR-27b inhibits PSCs proliferation and promotes PSCs differentiation. And the miR-27b target gene, MDFI, promotes PSCs proliferation and inhibits PSCs differentiation in vitro. Furthermore, interfering MDFI expression promotes mice muscle regeneration after injury. CONCLUSION: our results conclude that miR-27b promotes PSCs myogenesis by targeting MDFI. These results expand our understanding of muscle development mechanism in which miRNAs and genes work collaboratively in regulating skeletal muscle development. Furthermore, this finding has implications for obtaining the alternative treatments for patients with the muscle injury.

Integrated Analyses Reveal Overexpressed Notch1 Promoting Porcine Satellite Cells' Proliferation through Regulating the Cell Cycle.

Notch signaling as a conserved cell fate regulator is involved in the regulation of cell quiescence, proliferation, differentiation and postnatal tissue regeneration. However, how Notch signaling regulates porcine satellite cells (PSCs) has not been elucidated. We stably transfected Notch1 intracellular domain (N1ICD) into PSCs to analyze the gene expression profile and miRNA-seq. The analysis of the gene expression profile identified 295 differentially-expressed genes (DEGs) in proliferating-N1ICD PSCs (P-N1ICD) and nine DEGs on differentiating-N1ICD PSCs (D-N1ICD), compared with that in control groups (P-Control and D-Control, respectively). Analyzing the underlying function of DEGs showed that most of the upregulated DEGs enriched in P-N1ICD PSCs are related to the cell cycle. Forty-four and 12 known differentially-expressed miRNAs (DEMs) were identified in the P-N1ICD PSCs and D-N1ICD PSCs group, respectively. Furthermore, we constructed the gene-miRNA network of the DEGs and DEMs. In P-N1ICD PSCs, miR-125a, miR-125b, miR-10a-5p, ssc-miR-214, miR-423 and miR-149 are downregulated hub miRNAs, whose corresponding hub genes are marker of proliferation Ki-67 (MKI67) and nuclear receptor binding SET domain protein 2 (WHSC1). By contrast, miR-27a, miR-146a-5p and miR-221-3p are upregulated hub miRNAs, whose hub genes are RUNX1 translocation partner 1 (RUNX1T1) and fibroblast growth factor 2 (FGF2). All the hub miRNAs and genes are associated with cell proliferation. Quantitative RT-PCR results are consistent with the gene expression profile and miRNA-seq results. The results of our study provide valuable information for understanding the molecular mechanisms underlying Notch signaling in PSCs and skeletal muscle development.

MiR-34c represses muscle development by forming a regulatory loop with Notch1.

Since pork accounts for about 40% of global meat consumption, the pig is an important economic animal for meat production. Pig is also a useful medical model for humans due to its similarity in size and physiology. Understanding the mechanism of muscle development has great implication for animal breeding and human health. Previous studies showed porcine muscle satellite cells (PSCs) are important for postnatal skeletal muscle growth, and Notch1 signaling pathway and miRNAs regulate the skeletal muscle development. Notch1 signal pathway regulates the transcription of certain types of miRNAs which further affects target gene expression. However, the specific relationship between Notch1 and miRNAs during muscle development has not been established. We found miR-34c is decreased in PSCs overexpressed N1ICD. Through the overexpression and inhibition of mi-34c, we demonstrated that miR-34c inhibits PSCs proliferation and promotes PSCs differentiation. Using dual-luciferase reporter assay and Chromatin immunoprecipitation, we demonstrate there is a reciprocal regulatory loop between Notch1 and miR-34c. Furthermore, injection of miR-34c lentivirus into mice caused repression of gastrocnemius muscle development. In summary, our data revealed that miR-34c can form a regulatory loop with Notch1 to repress muscle development, and this result expands our understanding of muscle development mechanism.

Pig has no uncoupling protein 1.

Brown adipose tissue (BAT) is critical for mammal's survival in the cold environment. Uncoupling protein 1 (UCP1) is responsible for the non-shivering thermogenesis in the BAT. Pig is important economically as a meat-producing livestock. However, whether BAT or more precisely UCP1 protein exists in pig remains a controversy. The objective of this study was to ascertain whether pig has UCP1 protein. In this study, we used rapid amplification of cDNA ends (RACE) technique to obtain the UCP1 mRNA 3' end sequence, confirmed only exons 1 and 2 of the UCP1 gene are transcribed in the pig. Then we cloned the pig UCP1 gene exons 1 and 2, and expressed the UCP1 protein from the truncated pig gene using E. coli BL21. We used the expressed pig UCP1 protein as antigen for antibody production in a rabbit. We could not detect any UCP1 protein expression in different pig adipose tissues by the specific pig UCP1 antibody, while our antibody can detect the cloned pig UCP1 as well as the mice adipose UCP1 protein. This result shows although exons 1 and 2 of the pig UCP1 gene were transcribed but not translated in the pig adipose tissue. Furthermore, we detected no uncoupled respiration in the isolated pig adipocytes. Thus, these results unequivocally demonstrate that pig has no UCP1 protein. Our results have resolved the controversy of whether pigs have the brown adipose tissue.

Evaluation of Total Flavonoids, Myricetin, and Quercetin from Hovenia dulcis Thunb. As Inhibitors of α-Amylase and α-Glucosidase.

This study was designed to investigate the inhibition effect and mechanism of total flavonoids, myricetin and quercetin extracted from Hovenia dulcis Thunb. on α-amylase and α-glucosidase in order to explore the potential use of Hovenia flavonoids in alleviating postprandial hyperglycemia. The results demonstrate that total flavonoids, myricetin, and quercetin were effective inhibitors of α-amylase with IC50 values of 32.8, 662 and 770 µg ml-1, respectively. And all three were effective inhibitors of α-glucosidase with IC50 values of 8, 3 and 32 µg ml-1, respectively. Enzyme kinetics tests and Lineweaver-Burk results showed the inhibition effects of total flavonoids, myricetin and quercrtin on α-amylase were all reversible and competitive, and the effects on α-glucosidase were all reversible but non-competitive. This study revealed that Hovenia flavonoids, especially myricetin, are effective and promising functional foods in alleviating type 2 diabetes mellitus.

In-capillary formation of polymer/surfactant complexes-assisted reversed-migration micellar electrokinetic chromatography for facile analysis of neutral steroids.

In this study we developed a novel approach, using in-capillary formation of polymer/surfactant complexes (IPSC)-assisted reversed-migration MEKC (RM-MEKC), for the analysis of neutral steroids. This process involved two sequential events: in-capillary polymer/surfactant complexes formation during sample preconcentration, followed by IPSC separation. The procedure began with a polymer-filled capillary. Initially, on-line preconcentration of the sample was performed at the sample plug. Meanwhile, free surfactants migrated to interact with polymers, forming polymer-surfactant complexes. Analytes were then kinetically partitioned between the mixed phases (micelles and polymer-SDS complexes). Sodium dodecyl sulfate (SDS) and poly(N-isopropylacrylamide) (PNIPAAm) were employed as pseudo-stationary phases (PSPs). This system allowed the successful separation of five steroids (testosterone, hydrocortisone 21-acetate, dexamethasone, prednisolone, hydrocortisone) in acetate buffer and the determination of urinary free hydrocortisone; it also exhibited excellent performance for sample on-line concentration. The limit of detection for hydrocortisone was 20.98 ng/mL (R(2)=0.9995). The polymer size, concentrations, end-group charges, and SDS concentrations were evaluated. This IPSC/RM-MEKC system, which can be adopted in commercial CE instruments, is easy to operate, suitable for combination with several sample preconcentration options, sensitive, robust, and environmentally sustainable. We suspect that such systems might have potential applications in clinical analyses and in microanalytical devices.

Anti-inflammatory effect of spilanthol from Spilanthes acmella on murine macrophage by down-regulating LPS-induced inflammatory mediators.

Spilanthes acmella (Paracress), a common spice, has been administered as a traditional folk medicine for years to cure toothaches, stammering, and stomatitis. Previous studies have demonstrated its diuretic, antibacterial, and anti-inflammatory activities. However, the active compounds contributing to the anti-inflammatory effect have seldom been addressed. This study isolates the active compound, spilanthol, by a bioactivity-guided approach and indicates significant anti-inflammatory activity on lipopolysaccharide-activated murine macrophage model, RAW 264.7. The anti-inflammatory mechanism of paracress is also investigated. Extracts of S. acmella are obtained by extraction with 85% ethanol, followed by liquid partition against hexane, chloroform, ethyl acetate, and butanol. The ethyl acetate extract exhibits a stronger free radical scavenging capacity than other fractions do, as determined by DPPH and ABTS radical scavenging assays. The chloroform extract significantly inhibits nitric oxide production ( p < 0.01) and is selected for further fractionation to yield the active compound, spilanthol. The diminished levels of LPS-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) mRNA and protein expression support the postulation that spilanthol inhibits proinflammatory mediator production at the transcriptional and translational levels. Additionally, the LPS-stimulated IL-1beta, IL-6, and TNF-alpha productions are dose-dependently reduced by spilanthol. The LPS-induced phosphorylation of cytoplasmic inhibitor-kappaB and the nuclear NF-kappaB DNA binding activity are both restrained by spilanthol. Results of this study suggest that spilanthol, isolated from S. acmella, attenuates the LPS-induced inflammatory responses in murine RAW 264.7 macrophages partly due to the inactivation of NF-kappaB, which negatively regulates the production of proinflammatory mediators.