Modulation of the Tumor Metastatic Microenvironment and Multiple Signal Pathways by Prunella vulgaris in Human Hepatocellular Carcinoma.

Prunella vulgaris (PV) is a traditional Chinese medicine that has been used clinically for centuries in Asian countries to treat herpetic keratitis. In previous studies, PV was shown to suppress TPA-induced activation of MMP-9 and inhibit cell invasion and migration in hepatoma cell lines. However, the detailed molecular mechanism underlying these effects is still unclear. In this study, we investigated the mechanisms underlying PV-mediated inhibition of 12-O-Tetradecanoylphorbol-13-acetate (TPA)-induced cell invasion and inhibition of secreted and cytosolic MMP-9 production in human hepatoma cells (Huh-7 and HA22T). PV suppressed VEGF and MMP-9 transcription by inhibiting activator protein (AP)-1 and nuclear factor-[Formula: see text]B (NF-[Formula: see text]B) activity. PV suppressed TPA-induced AP-1 activity by inhibiting phosphorylation of the extracellular signal-related kinase (ERK), downregulating p38 signaling pathways, and suppressing TPA-induced inhibition of NF-[Formula: see text]B nuclear translocation through I[Formula: see text]B. PV suppressed TPA-induced activation of ERK/phosphatidylinositol-3-kinase/Akt upstream of NF-[Formula: see text]B and AP-1. These data suggest that PV modifies the metastatic microenvironment of hepatocellular carcinoma (HCC) by inhibiting multiple signal transduction pathways. PV thus may have the therapeutic potential to inhibit the migration and invasion of HCC and act as potential agent for systemic therapies.

Comparison of Newborn Hearing Screening in Well-Baby Nursery and NICU: A Study Applied to Reduce Referral Rate in NICU.

OBJECTIVES: To determine whether newborn hearing screening in a well-baby nursery (WBN) and neonatal intensive care unit (NICU) nursery: 1) meet three targeted, screening, referral, and diagnostic follow-up rates; 2) compare the average age of diagnosis for infants admitted to the WIN and NICU; and 3) determine prevalence of hearing loss in neonatal population; and 4) try to find a practical newborn hearing screening time algorithm to reduce refer rate in NICU. MATERIALS AND METHODS: It examined 15,624 newborns in the WBN (13,676) and NICU (1948) screened for congenital HL using AABR. The variables analyzed in it were the screening rate, referral rate, follow-up rate, diagnostic rate and diagnostic age, prevalence rate, degrees of congenital bilateral HL. The study was approved by the hospital's institutional review board (13MMHISO23). RESULTS: The screening rates were 99.8% and 99.6% in the WBN and NICU groups, respectively, without significant difference. The referral rates were 0.7% and 2.8% in the WBN and NICU groups, with significant difference. Furthermore, the diagnostic follow-up rates were 76.7% and 89.1% in the WBN and NICU groups, without significant difference. The average initial diagnostic ages were 1.9 months and 3.8 months in the WBN and NICU groups, with significant difference. The prevalence of congenital bilateral hearing loss were 0.27% and 1.6% in the WBN and NICU groups, with significant difference. CONCLUSION: The screening, referral and follow-up rate in the WBN and NICU groups were equivalent to the quality indicators. For NICU group, screening and diagnostic follow up were performed later than those in WBN group; however the lower referral rate in our NICU group was successfully achieved in this study and can be applied clinically. The prevalence of congenital bilateral hearing loss was higher in the NICU group than in the WBN group.

Naringenin suppresses TPA-induced tumor invasion by suppressing multiple signal transduction pathways in human hepatocellular carcinoma cells.

Naringenin, a common dietary flavonoid abundantly present in fruits and vegetables, is believed to possess strong anti-proliferative properties and the ability to induce apoptosis in hepatoma cell lines. However, there are no reports describing its effects on the invasion and metastasis of hepatoma cell lines, and the detailed molecular mechanisms of its effects are still unclear. In this study, we investigated the mechanisms underlying naringenin-mediated inhibition of 12-O-Tetradecanoylphorbol-13-acetate (TPA)-induced cell invasion and inhibition of secreted and cytosolic MMP-9 production in human hepatoma cells (HepG2, Huh-7, and HA22T) and murine embryonic liver cells (BNL CL2). Naringenin suppressed MMP-9 transcription by inhibiting activator protein (AP)-1 and nuclear factor-κB (NF-κB) activity. It suppressed TPA-induced AP-1 activity through inhibiting the phosphorylation of the extracellular signal-related kinase (ERK) and c-Jun N-terminal kinase (JNK) signaling pathways, and it suppressed TPA-induced inhibition of NF-κB nuclear translocation through IκB. Additionally, it suppressed TPA-induced activation of ERK/phosphatidylinositol 3-kinase/Akt upstream of NF-κB and AP-1. These data suggest that naringenin suppresses the invasiveness and metastatic potential of hepatocellular carcinoma (HCC) by inhibiting multiple signal transduction pathways.

Protective and immunomodulatory effect of flos Lonicerae japonicae by augmenting IL-10 expression in a murine model of acute lung inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Lonicerae japonicae has shown antibacterial, anti-inflammatory, antipyretic, antioxidant, antiviral, and protective effects in animal models of chronic obstructive pulmonary disease. AIM OF THE STUDY: To investigate the effects of Flos L. japonicae (FLJ), a traditional Chinese medicinal herb, on acute lung inflammation induced by lipopolysaccharide (LPS), in vivo, using a murine model. MATERIALS AND METHODS: Thirty 6-week-old female BALB/c mice were challenged with intratracheal LPS before treatment with FLJ or vehicle. FLJ was examined for its capacity to influence an LPS-induced increase in IL-10 and decrease in TNF-α, IL-1ß and IL-6 as measured in murine bronchoalveolar lavage (BALF). RESULTS: FLJ increased nuclear Sp1 binding activity thereby enhancing the expression of IL-10 and decreased nuclear NF-κB binding activities thereby inhibiting the expression of TNF-α, IL-1ß and IL-6 in the lung. The up-regulation of Sp1 activity by FLJ was through incremental phosphorylation of ERK. By contrast, the down-regulation of NF-κB activity by FLJ was through suppression of the phosphorylation of IκB, p38, and JNK. CONCLUSIONS: We demonstrated FLJ has protective activity against LPS-induced lung inflammatory cytokine release. Anti-inflammatory cytokine, IL-10, may prove beneficial in the treatment of endotoxin-associated lung inflammation.

Genistein inhibits tumor invasion by suppressing multiple signal transduction pathways in human hepatocellular carcinoma cells.

BACKGROUND: Genistein (Gen) exhibits anti-mutagenic and anti-metastatic activities in hepatoma cell lines. Gen has suppressive effects on tumor growth and angiogenesis in nude mice. Gen suppresses the enzymatic activity of matrix metalloproteinase (MMP)-9; however, the mechanism underlying its anti-invasive activity on hepatocellular carcinoma (HCC) cells is unclear. METHODS: In this study, the possible mechanisms underlying Gen-mediated reduction of 12-O-Tetradecanoylphorbol-13-acetate (TPA)-induced cell invasion and inhibition of secreted and cytosolic MMP-9 production in human hepatoma cells (HepG2, Huh-7, and HA22T) and murine embryonic liver cells (BNL CL2) were investigated. RESULTS: Gen suppressed MMP-9 transcription by inhibiting activator protein (AP)-1 and nuclear factor-κ B (NF-κB) activity. Gen suppressed TPA-induced AP-1 activity through inhibitory phosphorylation of extracellular signal-related kinase (ERK) and c-Jun N-terminal kinase (JNK) signaling pathways, and TPA-stimulated inhibition of NF-κB nuclear translocation through IκB inhibitory signaling pathways. Moreover, Gen suppressed TPA-induced activation of ERK/phosphatidylinositol 3-kinase/Akt upstream of NF-κB and AP-1. CONCLUSIONS: Gen and its inhibition of multiple signal transduction pathways can control the invasiveness and metastatic potential of HCC.

A new approach to sound source segregation.

We rely critically on our ability to 'hear out' (segregate) individual sound sources in a mixture. Yet, despite its importance, little is known regarding this -ability. Perturbation analysis is a psychophysical method that has been successfully applied to related problems in vision [Murray, R.F. 2011. J. of Vision 11, 1-25]. Here the approach is adapted to audition. The application proceeds in three stages: First, simple speech and environmental sounds are synthesized according to a generative model of the sound--producing source. Second, listener decision strategy in segregating target from non--target (noise) sources is determined from decision weights (regression coefficients) relating listener judgments regarding the target to lawful perturbations in acoustic parameters, as dictated by the generative model. Third, factors limiting segregation are identified by comparing the obtained weights and residuals to those of a maximum-likelihood (ML) observer that optimizes segregation based on the equations of motion of the generating source. Here, the approach is applied to test between the two major models of sound source segregation; target enhancement versus noise cancellation. The results indicate a tendency of noise segregation to preempt target enhancement when the noise source is unchanging. However, the results also show individual differences in segregation strategy that are not evident in the measures of performance accuracy alone.

Auditory discrimination of force of impact.

The auditory discrimination of force of impact was measured for three groups of listeners using sounds synthesized according to first-order equations of motion for the homogenous, isotropic bar [Morse and Ingard (1968). Theoretical Acoustics pp. 175-191]. The three groups were professional percussionists, nonmusicians, and individuals recruited from the general population without regard to musical background. In the two-interval, forced-choice procedure, listeners chose the sound corresponding to the greater force of impact as the length of the bar varied from one presentation to the next. From the equations of motion, a maximum-likelihood test for the task was determined to be of the form Δlog A + αΔ log f > 0, where A and f are the amplitude and frequency of any one partial and α = 0.5. Relative decision weights on Δ log f were obtained from the trial-by-trial responses of listeners and compared to α. Percussionists generally outperformed the other groups; however, the obtained decision weights of all listeners deviated significantly from α and showed variability within groups far in excess of the variability associated with replication. Providing correct feedback after each trial had little effect on the decision weights. The variability in these measures was comparable to that seen in studies involving the auditory discrimination of other source attributes.

Target enhancement and noise cancellation in the identification of a rudimentary sound source in noise.

Perturbation analysis was used to determine the relative contribution of target enhancement and noise cancellation in the identification of rudimentary sound source in noise. In a two-interval, forced-choice procedure, listeners identified the impact sound produced by the larger of two stretched membranes as target. The noise on each presentation was the impact sound of a variable-sized plate. For four of five listeners, the relative weights on the noise were positive indicating enhancement, and for the remaining listeners, they were negative indicating cancellation. The results underscore the difficulty with evaluating models of masking solely in terms of measures of performance accuracy.

A method for evaluating the relation between sound source segregation and masking.

Sound source segregation refers to the ability to hear as separate entities two or more sound sources comprising a mixture. Masking refers to the ability of one sound to make another sound difficult to hear. Often in studies, masking is assumed to result from a failure of segregation, but this assumption may not always be correct. Here a method is offered to identify the relation between masking and sound source segregation in studies and an example is given of its application.

The inhibitory effect of hesperidin on tumor cell invasiveness occurs via suppression of activator protein 1 and nuclear factor-kappaB in human hepatocellular carcinoma cells.

Hesperidin (HES) has been reported to exhibit anti-invasive and antimetastatic activities by suppressing the enzymatic activity of matrix metalloproteinase-9 (MMP-9). However, the underlying mechanism of anti-invasive activity remains unclear so far. First, we suggest that the expression of MMP-9 by TPA involves phosphorylation of IKK, p38, and PKC in hepG2. We also demonstrate that hesperidin reduced 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cell invasion and inhibited the secreted and cytosolic MMP-9 forms in HepG2 cells. Hesperidin significantly suppressed the TPA-induced the mRNA level of MMP-9. Hesperidin suppressed MMP-9 transcription by inhibiting nuclear factor-kappaB (NF-kappaB) and Activator protein 1 (AP-1) activity. Hesperidin suppressed TPA-stimulated NF-kappaB translocation into the nucleus through IkappaB inhibitory signaling pathways and also inhibited TPA-induced AP-1 activity by the inhibitory phosphorylation of p38 kinase and c-Jun N-terminal kinase (JNK) signaling pathways. In conclusion, Hesperidin might be a potent antiinvasive agent that suppresses the MMP-9 enzymatic activity via NF-kappaB an AP-1 signaling pathway.

Xia-bai-san inhibits lipopolysaccharide-induced activation of intercellular adhesion molecule-1 and nuclear factor-kappa B in human lung cells.

UNLABELLED: Xia-bai-san (XBS) is a traditional Chinese medicine that has been used clinically for centuries in Asian countries to treat some types of common cold and asthma-like diseases similar to infantile pneumonia and childhood bronchitis. In previous studies, XBS was found to suppress the inflammatory process induced in lungs of mice treated with lipopolysaccharide (LPS). PURPOSE: The present study was undertaken to examine the effects of XBS on LPS-inducible production of inflammatory cytokines, up-regulation of intercellular cell adhesion molecule-1 (ICAM-1), and activation of nuclear factor NF-kappaB in cultured human lung cells. PRINCIPAL RESULTS: Extracts of four raw herbs (Cortex mori, Cortex lycii, Radix glycyrrhizae, and Fructus oryzae) were used to prepare the decoction. XBS decreased the histological damage and up-regulation of ICAM-1 observed in lungs of mice treated with lipopolysaccharide (LPS). In cultured human pulmonary epithelial A549 cells, XBS and its components Morus alba and Glycyrrhiza uralensis suppressed the up-regulation of IL-8 and ICAM-1 in response to LPS. Production of TNF-alpha, IL-1beta, IL-6 and IL-8 by LPS-treated human THP-1 monomyelocytes was also suppressed by XBS. A549 cells expressed ICAM-1 in response to medium from LPS-treated THP-1 cells; expression was decreased by XBS. The adhesion of THP-1 cells to LPS-treated A549 cells were inhibited in the presence of XBS. Activation of NF-kappaB by LPS in A549 cells was suppressed by XBS, Morus alba, and Glycyrrhiza uralensis through inhibition of IkappaB phosphorylation; the concentrations at which suppression occurred were identical to those at which production of inflammatory cytokines and up-regulation of ICAM-1 were inhibited. MAJOR CONCLUSIONS: These findings support the hypothesis that XBS, Morus alba, and Glycyrrhiza uralensis inhibit the inflammatory process in lung tissue through suppression of the IkappaB signaling pathway. XBS may prove helpful in the management of asthma, various allergic disorders, sepsis, or any other condition associated with pulmonary inflammation.

Hesperidin inhibited acetaldehyde-induced matrix metalloproteinase-9 gene expression in human hepatocellular carcinoma cells.

Previous studies have revealed that acetaldehyde-induced cell invasion and matrix metalloproteinase-9 (MMP-9) activation and are directly involved in hepatic tumorigenesis and metastasis. Acetaldehyde is an important substance for tumor regression. We designed this study to aid in the development of powerful anti-cancer drugs with specific tumor regression and anti-metastatic potentials. Optimal drugs should possess both specific MMP-9 enzyme and gene transcriptional activities at the molecular level. Hesperidin, a flavonoid present in fruits and vegetables, possess anti-inflammatory and chemopreventive activities. Hesperidin suppressed acetaldehyde-induced cell invasion and inhibited the secreted and cytosolic MMP-9 forms in HepG2 cells with acetaldehyde. Hesperidin suppressed acetaldehyde-induced MMP-9 expression through the inhibition of nuclear factor-kappaB (NF-kappaB) and AP-1, and suppressed acetaldehyde-stimulated NF-kappaB translocation into the nucleus through IkappaB inhibitory signaling pathways. Hesperidin also inhibited acetaldehyde-induced AP-1 activity by the inhibitory phosphorylation of p38 kinase and c-Jun N-terminal kinase (JNK) signaling pathways. Results from our study revealed that hesperidin suppressed both acetaldehyde-activated NF-kappaB and activator protein 1 (AP-1) activity by IkappaB, JNK, and p38 signaling pathways. This resulted in the reduction of MMP-9 expression, secretion, and hepatocarcinoma cellular invasion. Our result confirmed the therapeutic potential of hesperidin an anti-metastatic and its involvement in the acetaldehyde-induced cell invasiveness of hepatocellular carcinoma in alcoholic patients.

Level dominance in sound source identification.

Impact sounds were synthesized according to standard textbook equations given for the motion of simply supported, metal plates. In a two-interval, forced-choice procedure, highly practiced listeners identified from these sounds a predefined class of target plates based on their particular material and geometric properties. The effects of two factors on identification were examined: the relative level of partials comprising the sounds and the relative amount of information (given as the difference in d(')) each partial provided for identification. In different conditions one factor was fixed while the other either increased or decreased with frequency. The effect on listener identification in each case was determined from a logistic discriminant analysis of trial-by-trial responses, yielding a vector of listener decision weights on the frequency and decay of individual partials. The weights increased proportionally with relative level, but were largely uninfluenced by relative information content--a result exactly opposite to that expected from a maximum-likelihood observer. The dominant effect of relative level was replicated for other sound sources (clamped bars and stretched membranes) and was not diminished by randomizing the relative level of partials across trials. The results are taken to underscore the importance of relative level in the identification of rudimentary sound sources.

Individual differences in source identification from synthesized impact sounds.

Impact sounds were synthesized according to standard textbook equations given for the motion of freely vibrating membranes, bars, and plates. In a two-interval, forced-choice procedure, highly practiced listeners identified from these sounds predefined target sources based on their material and size, the hardness of the striking mallet, and the presence or absence of light damping applied to the center of the source. Listener decision strategy in each case was determined from a discriminant analysis of trial-by-trial responses resulting in a vector of regression weights given to different acoustic parameters. The analysis revealed significant differences in decision strategy across listeners within identification task, but similarity in decision strategy within listeners across variations in task. Only when the acoustic information for identification was highly constrained (identification of damping) did listeners adopt similar decision strategies approaching that of an ideal observer. Despite the large individual differences in decision strategy, identification accuracy was, in most cases, similar across listeners. Where there were differences in identification accuracy the differences appeared largely related to differences in internal noise and not decision strategy. The results are generally comparable to those obtained for the discrimination of arbitrary tone patterns.

The immunomodulation of endotoxin-induced acute lung injury by hesperidin in vivo and in vitro.

To investigate the modulation of lung local immune responses of hesperidin (HES) on the acute lung inflammation induced by LPS in vivo. Mice were challenged with intratracheal lipopolysaccharide (100 microg) 30 min before with treatment hesperidin (200 mg/kg oral administration) or vehicle. After 4 and 24 h, bronchoalveolar lavage fluid was obtained to measure proinflammatory (TNF-alpha, IL-1 beta, IL-6), anti-inflammatory (IL-10, IL-4, IL-12) cytokines, chemokines (KC, MCP-1 and MIP-2), total cell counts, nitric oxide production, and proteins. Lung histology was performed in inflated-fixed lungs. Hesperidin downregulate the LPS-induced expression of TNF-alpha, IL-1 beta, IL-6, KC, MIP-2, MCP-1, and IL-12. It also enhanced the production of IL-4, IL-10. Total leukocyte counts; nitric oxide production, iNOS expression, and proteins were significantly decreased by hesperidin. In vitro, HES suppressed the expression of IL-8 on A549 cells and THP-1 cells, the expression of TNF-alpha, IL-1 beta, and IL-6 on THP-1 cells, the expression of ICAM-1 and VCAM-1 on A549 cells which effect cell adhesion function. The suppression of those molecules is controlled by NF-kappaB and AP-1, which are activated by I kappa B and MAPK pathways. HES inhibits those pathways, thereby suppressing the expression of IL-8, TNFalpha, IL-1 beta, IL-6, IL-12, ICAM-1 and VCAM-1. This study indicates that HES had a markedly immunomodulatory effect in a clinically relevant model of ARDS. Nevertheless, further investigations are required to determine the potential clinical usefulness of HES in the adjunctive therapy of ARDS.

Protective and immunomodulatory effect of Gingyo-san in a murine model of acute lung inflammation.

To investigate the effects of Gingyo-san (GGS), the traditional Chinese medicinal formula, on the acute lung inflammation induced by LPS in vivo, mice were challenged with intratracheal LPS before treatment with GGS or vehicle. In lung morphology, GGS reduced the infiltration of activated polymorphonuclear neutrophils in the airways, decreased pulmonary edema, reduced nitrosative stress, and improved lung morphology. ELISA or RT-PCR detected the expression of cytokines in BALF and lung tissue. The mechanism of these benefits by treatment with GGS including attenuating expression TNFalpha, IL-1 beta, IL-6, KC, MCP-1, MIP-2, iNOS, and activation of nuclear factor (NF-kappaB and AP-1) in BALF and lung tissue. Particularly, GGS also enhanced the anti-inflammatory cytokine (IL-10) and limited the acute lung inflammation. Therefore, its protection activity against LPS-induced lung inflammatory mediators release might be beneficial in the treatment of endotoxin-associated inflammation.

Microdermabrasion as a novel tool to enhance drug delivery via the skin: an animal study.

BACKGROUND: Microdermabrasion is a widely performed skin rejuvenation procedure. It can partly ablate and homogenize the stratum corneum (SC) layers. OBJECTIVE: The effect of microdermabrasion treatment on the skin permeation of hydrophilic and lipophilic drugs was examined in this study. METHODS: 5-Fluorouracil (5-FU) and clobetasol 17-propionate were used as the hydrophilic and lipophilic permeants, respectively. In vitro skin delivery using porcine skin and in vivo topical application employing nude mouse as the animal model were both used to examine the effect of microdermabrasion. The vacuum pressures used in this study (15-25 cmHg) were much lower than those used for therapeutic purposes. RESULTS: The 5-FU permeation across microdermabrasion-treated skin was 8- to 24-fold higher than that across intact skin and depended on differences in treatment pressure and duration. An intensity of 15 cmHg for 10 seconds showed the greatest enhancement of 5-FU delivery via the skin. In contrast to the results for 5-FU, microdermabrasion reduced the skin permeation and deposition of topically applied clobetasol. The partitioning effect of clobetasol from the vehicle to the SC may have predominated this result. Microdermabrasion also enhanced the skin delivery of the hydrophilic 5-aminolevulinic acid (ALA). Confocal laser scanning microscopy (CLSM) of microdermabrasion-treated skin revealed intense red fluorescence of ALA-transformed protoporphyrin (PpIX) within the epidermis and upper dermis. CONCLUSIONS: Microdermabrasion can improve the skin permeation of hydrophilic molecules.