Soil fertility and nutrient-related plant functional traits are in general only moderately related, hindering the progress in trait-based prediction models of vegetation patterns. Although the relationships may have been obscured by suboptimal choices in how soil fertility is expressed, there has never been a systematic investigation into the suitability of fertility measures. This study, therefore, examined the effect of different soil fertility measures on the strength of fertility–trait relationships in 134 natural plant communities. In particular, for eight plot-mean traits we examined (1) whether different elements (N or P) have contrasting or shared influences, (2) which timescale of fertility measures  has better predictive power, and (3) if integrated fertility measures explain trait variation better than individual fertility measures. Soil N and P had large mutual effects on leaf nutrient concentrations, whereas they had element-specific effects on traits related to species composition (e.g. Grime's CSR strategy). The timescale of fertility measures only had a minor impact on fertility–trait relationships. Two integrated fertility measures  were related significantly to most plant traits, but were not better in explaining trait variation than individual fertility measures. Using all fertility measures together, between-site variations of plant traits were explained only moderately for some traits (e.g. 33% for leaf N concentrations) but largely for others (e.g. 66% for whole-canopy P concentration). The moderate relationships were probably due to complex regulation mechanisms of fertility on traits, rather than to a wrong choice of fertility measures. We identified both mutual (i.e. shared) and divergent (i.e. element-specific and stoichiometric) effects of soil N and P on traits, implying the importance of explicitly considering the roles of different elements to properly interpret fertility–trait relationships. In the short term, soil nutrients induce plastic responses of individual plants . In the longer run, soil nutrients affect the species composition of a plant community, because each species has evolved to adapt to certain environments and therefore has contrasting requirements for nutrients. How and why plant community composition changes from fertile to unfertile soil has been a central concern of ecologists for many decades. A general trend derived from qualitative studies is that species with a rapid growth strategy dominate in fertile soils, whereas species with a conservative strategy dominate in infertile soils  or explicitly  reflected in the way the measure is expressed. Measures from chemical soil bulk analysis at a certain moment, such as pool size and extractable amount of nutrients, provide only partial information about the nutrient availability for plants over a long time span Moreover, since plants use soil nutrients in a multidimensional manner , certain combinations of soil fertility measures may better reflect plant trait variability. Hence, an aggregated measure derived from multiple fertility measures may better explain trait variability than a single fertility measure.This study aims to investigate whether the relationships between soil fertility and nutrient-related plant traits can be improved by analyzing the mutual and divergent impacts of different soil fertility measures. To this aim, we tested (1) whether nutrient elements in soil (N or P) have specific or shared impacts on plot-mean plant traits , (2) whether the timescale of a soil fertility measure is important in explaining plot-mean plant traits, and (3) whether use of an integrated soil fertility measure improves the fertility vs. trait relationships. Subsequently, we infer from our findings which fertility measure or combinations of fertility measures are the best suited to describe among-plots trait variations. Throughout the paper, we considered vascular plants only, since non-vascular species (i.e. mosses and lichens) do not directly take up nutrients from soils and are thus less relevant in the context of soil fertility–trait relationships.We obtained permissions for soil and vegetation sampling in Hoge Veluwe national park , Zuid-Kennemerland national park , and nature reserves owned by Staatsbosbeheer.2 for herbaceous, 4, 9, or 25 m2 for shrub (depending on the size of the woody species), and 100 m2 for forest stands. Soil cores of 10 cm depth (datasets 1 and 3) or 15 cm depth (dataset 2) were taken within or next to the plot. Large roots were removed from the soil cores. For datasets 1 and 3, several soil cores were mixed to make a composite soil to eliminate the effects of local soil heterogeneity. In the peak growing season (July or August), above-ground standing biomass of vascular plants (datasets 1 and 2) or leaves of dominant vascular species (dataset 3) were harvested.We selected 36 sites in the Hoge Veluwe and Zuid-Kennemerland national parks and nature reserves owned by Staatsbosbeheer (dataset 1). Additionally, we used 47 sites of Olde Venterink et al. 3 + N–NH4, mg N kg−1 soil) and dissolved P  were measured to indicate the short-term availability of N and P, respectively. Two different extraction methods were used to measure dissolved P: ammonium lactate-acetic acid (ALA) extraction and the Olsen extraction. Therefore, we converted the values estimated with the ALA extraction to be comparable to those with Olsen extraction, using an empirically derived relationship Dissolved N (N–NO−1 soil week−1). Because denitrification in wet soil cores could have caused a considerable loss of N from the incubation tubes As a longer-term indication of N availability, we measured in-situ N mineralization rates in the mid growing season (‘summer Nmin’) for 6 weeks (datasets 1 and 3) or 8 weeks (dataset 2)  and for the preceding five years (from September five years before the sampling year to August of the sampling year) . Transformation processes of mineralized N and P  were not considered.Additionally, to estimate nutrient availability for much longer terms, we used a modified version of a SOM model, CENTURY Finally, as very rough measures of soil fertility in the long time span, we used soil total N and P , soil N:C ratio, and soil P:C ratio.−1) were determined for dataset 3 only  for datasets 1 and 2  was sampled to determine whole-canopy N and P concentrations in each plot  to 3 (prevailing at nutrient rich sites) nut values were computed as arithmetic means of IVnut for each site, rather than weighted means of IVnut based on species abundance, as the former was shown to be sufficient for this trait with ordinal-scale values nut value was not available, were excluded from the calculation of plot-mean IVnut values. The trait coverage  ranged from 76% to 100% (median 94%).IVThe CSR scheme represents the adaptive strategy of plant species along gradients of resource availability, stress, and disturbance, expressed with three axes of primary components: C (‘Competitors’), S (‘Stress tolerators’), and R . Each species can be classified into one out of 19 classes with different combinations of strategy components, e.g. C, SR/R, or CSR. We retrieved CSR strategies from Hunt et al. t  into unique and shared effects of the two groups of predictor variables; i.e. to N-related fertility measures  and P-related fertility measures  Tt regressed by X, where X is either N, P, or N&P. In order to correct for the different number of fertility measures within each group, we used adjusted R2, R2Tt|X) adj adj( − R2Tt|P) adj adj( − R2Tt|N) adj adj( + R2Tt|P) adj( − R2Tt|N&P) adj adj-boot,i( − R2Tt|P) adj-boot,i or above zero (when the median of Di was negative).Additionally, we tested if N-related fertility measures explain significantly more variance in plot-mean traits than P-related fertility measures, and vice versa, following the bootstrapping method described in k fertility measures with different timescales (predictor variables), to quantify the independent contribution of a fertility measure S to the explained variance of a trait without being confounded by the other k-1 fertility measures. The hierarchical partitioning method computes the increase in goodness-of-fit when S is added to the model (in our case: a linear multivariate regression model of a trait regressed by fertility measures) compared to the model without S, and averages the increase over all possible models that include S as a predictor variable (i.e. 2k models). In this way, the variation of a trait explained by S is divided into an independent effect of S and joint effect of S with other fertility measures. Negative values of a joint effect mean that the interactive effects of S and the other fertility measures on the trait are suppressive, rather than enhancing. An advantage of using the hierarchical partitioning over a one-model approach is that the averaging eliminates the problem of multicollinearity among predictor variables k = pN or k = pP), so that interactions of N and P do not obscure the effects of timescale. We used R2 as the goodness-of-fit measure of the models. The analysis was performed in R In order to examine the most relevant timescale of soil fertility for explaining the variation of plant traits, we used the hierarchical partitioning method Foe each fertility measures, the statistical significance of their independent contribution to a plant trait was tested by randomizing the pairs of trait and fertility values for 1000 times p<0.05) different magnitudes of independent effects.In addition, independent contributions were compared among fertility measures with different timescales by means of bootstrapping in R Since the soil fertility measures were strongly correlated, we extracted the main axes of variation by a Principal Component Analysis (PCA) in R p = 0.42 for LNC, p = 0.30 for LPC). P concentrations at the whole-canopy level were more strongly related to soil fertility measures than those on a leaf level .Bivariate correlations between plant traits and soil fertility measures were often significant . All soinut was explained by soil fertility measures (p = 0.33).55.5% of the variance in IVmeasures , of whicp = 0.057) larger than that of soil P measures, whereas for the C and S components, the contribution of soil P measures was considerably  larger than that of soil N measures.Of the three components of the CSR strategy, soil fertility measures explained the variance in the S component to a largest extent 50.7%, , followep>0.05) different among fertility measures with different timescales  except between 5yr Pmin and annual Pmin  different among fertility measures with different timescales except between 5yr Pmin and annual Pmin  independent effects than shorter-term measures   .PCA analysis extracted major axes of variation in soil fertility measures. The first axis, explaining 59.8% of the variance, was related to overall nutrient availability of a site (i.e. high in both soil N and P), with the negative axis values representing fertile conditions . The secnut, the S component, and the R component (p<0.05) effects for WPC, IVnut, and the S component ; this indicates that the two axes had complementary rather than redundant effects on these traits.Most plant traits, except WNC and the R component, were significantly related to PCA axis 1 . PCA axiomponent . MultiplIn line with previous studies There are several possible mechanisms that explain why LNC is related to both soil N and soil P (rather than only to soil N) and why LPC is related to both soil N and soil P (rather than only to soil P). Although leaf-level nutrient concentrations change plastically with changing concentrations of that element in the soil In contrast, the nutrient concentrations on a whole-canopy level (WNC and WPC) reflect the availability of single elements more strongly than on a leaf level, because plants store excess elements in non-leaf organs. This is reflected in a much larger variation in N:P ratio in non-leaf organs than in leaves nut, the unique effects of single elements contributed to most of the explained variance, although the shared effect of soil N and P also accounted for a large part of the variance. For the CSR strategy, most of the explained variance was related to the unique effects of single elements, rather than to the shared effects of soil N and P. Here, the effects of N and P were highly different, indicating a stoichiometric control of soil N and P on the CSR strategy of vegetation. The unique effects of soil P were larger than those of soil N for the C and S components, in accordance with the findings of Ceulemans et al. For IVp = 0.094], small shared effects on CSR strategy) remained. When only P-limited plots were considered, the contributions of soil P remained mostly unchanged or even decreased for some traits . It is particularly notable that the stronger determinants for stress tolerators were N-related fertility measures in N-limited plots (p<0.05) and P-related fertility measures in P-limited plots (p = 0.079) . Furthermore, signs of correlations between soil N measures and integrative plant traits  reversed in P-limited plots compared to N-limited plots for most cases  whether adding extra measurements of soil P availability improved the fertility–trait relationships . Neither2 values in nut, S component, R component). Similarly, including the type of nutrient limitation of plants (i.e. N- or P-limited) improved the relationship between overall fertility and several plant traits  , was related to almost all plant traits. However, the explanatory power of the PCA axis 1  is a prerequisite for improving fertility–trait relationships. Note that the N:P stoichiometry effect cannot be assessed by individual fertility measures alone, but it can be explicitly tested by integrated fertility measures (i.e. PCA axis 2 in our case). This suggests the appropriateness of using integrated fertility measures as a starting point to explore which aspect of soil fertility has a relevant effect on the specific plant trait.nut, respectively. Reasonably good relationships between IVnut or equivalent  and a single soil fertility measure have also been observed before  is primarily associated with disturbance rather than soil fertility In conclusion, our study showed that among-site variations in nutrient-related plant traits are consistently, although moderately for some traits, related to soil fertility measures. Whether a trait has only a moderate relationship depends on the mechanism through which soil fertility and other factors regulate the trait variation. The timescale of the fertility measure has only negligible or minor effects on fertility–trait relationships, whereas the mutual and/or stoichiometric effects of N and P should be considered to improve the relationships. Since the relative importance of soil fertility measures is different among plant traits, a scan of integrated fertility measures will facilitate identification of influential fertility measures (or groups of fertility measures) for each specific plant trait separately.Table S1Correlations among soil fertility measures.(DOCX)Click here for additional data file.Table S2Correlations between plot-mean plant traits and soil fertility measures.(DOCX)Click here for additional data file.Table S3Comparison of 1000x bootstrapped independent effects among soil fertility measures.(DOCX)Click here for additional data file.Appendix S1Testing the effects of using two extraction methods for dissolved P.(DOCX)Click here for additional data file.Appendix S2Testing the effects of using Olsen-P extraction for acid soils.(DOCX)Click here for additional data file.Appendix S3Fertility–trait relationships in N- and P-limited ecosystems.(DOCX)Click here for additional data file.Appendix S4Testing additional soil P measures for a subset of dataset.(DOCX)Click here for additional data file.


HTLV-1 is endemic in parts of Africa and the highest prevalence (5%) in West Africa has been reported in Caio, a rural area in Guinea-Bissau. It is unknown which HTLV-1 subtypes are present in this community.To compare local HTLV-1 sequence variation and to describe the phylogeny of the virus in the Caio population.The complete Long Terminal Repeat (LTR) region of HTLV-1 samples from children and adults was sequenced. Socio-demographic data were obtained from routine census data and from interviews. Phylogenetic analyses were performed to characterize the viruses.The complete 800-bp LTR sequences from 38 HLTV-1 infected adults and 8 children were obtained. Strikingly, sequences from many unrelated individuals showed 100% nucleotide identity . Phylogenetic analysis demonstrated that 45 of the sequences belonged to the cosmopolitan subtype A, subgroup D. One sequence, Caio4046, was divergent and formed a significant cluster with the STLV-1 sequences from the Central African Republic and Senegal and with a recently identified HTLV-1 strain from a Cameroonian hunter (tentatively designated subtype G) [Markedly conserved HTLV-1 strains of the cosmopolitan subtype A, subgroup D, predominate in this rural community. However, HTLV-1 subtype G is also present. This subtype has not been described before in a non-hunter or in West Africa and suggests this subtype may be more widespread than previously thought. Additional sequencing can give more information about the frequency of this subtype in Caio.


Background. Baduanjin is a traditional Chinese qigong that has been practiced for a long time in China as a mind-body exercise in community elderly populations. The objective of this study was to qualitatively evaluate the perceived benefit of regular Baduanjin qigong in community elders. Methods. A total of 20 participants who had completed the 12-week Baduanjin qigong training were interviewed regarding their perceived effect on physical and psychological health and whether Baduanjin qigong was suitable for the elderly. Results. Almost all participants agreed that Baduanjin qigong could promote their multisystem or organ functions , increase their immunity, make their bodies relax, and improve their mood and confidence. Most of the participants also agreed that Baduanjin qigong was appropriate for elderly individuals. Few individuals felt bored because of an hour Baduanjin training each day. Conclusions. The findings suggest that regular Baduanjin qigong may be potentially helpful to promote the overall physical and psychological health of elderly community populations and may be useful and feasible as a body-mind exercise in the health promotion in the elderly community populations. Baduanjin qigong has a long history in China as a comprehensive mind-body practice that involves physical postures and movement, breathing exercise, and relaxation, and it has been recommended for general use in the community by the Chinese Health Qigong Association , 2. Its Moreover, the majority of previous studies have only considered the variation between Baduanjin qigong and physical or psychological outcomes by quantitative measurement. Little information is available on the relationship between Baduanjin qigong and the practitioner's subjective experiences, which might present new outcomes. As a complex holistic “mind-body exercise” with the obvious characteristics of traditional Chinese medicine –15, BaduParticipants in the current qualitative study were recruited from those who had completed the Baduanjin qigong training program that took part in a RCT . In the Because of personal and time constraints, purposive samples of twenty participants  with median age of 60 years and range from 50 to 69 years from Baduanjin qigong group were selected to participate in the semistructured interviews by using convenient sampling method. Seventeen of them had low levels of education , and three participants had a college or higher level of education. Most of them had hypertension (15/20) or dyslipidemia (17/20); 13 participants were overweight. The participant characteristics were shown in Data were collected from August 1 to September 30, 2014, after the 12-week Baduanjin qigong training program. Two of the authors (Qianying Fang and Bai Chen) created an interview agenda mainly designed to explore subjective experiences of Baduanjin qigong for body-mind health, including experiences with Baduanjin qigong training, body and mind changes associated with Baduanjin qigong, and suitability of Baduanjin qigong for elders in community (Appendix). Each individual interview was conducted face to face by the authors (Qianying Fang and Bai Chen) at a quiet room of the participant's home. The interview scope gradually extended to focus on two main topics: (1) body and mind changes of the individual after the Baduanjin qigong training and (2) whether Baduanjin qigong is suitable for the elderly and why. The participants were encouraged to speak freely and to express their own views. Each interview was audio-recorded with participant permission and took at least 40 minutes. Additional data were also collected after each interview in the form of interviews' written notes and personal observations. The tape-recorded interviews were transcribed verbatim into Chinese, and the transcribed text was randomly double-checked.According to the qualitative content analysis, as described by Graneheim and Lundman 2004) [04 [20], The study was approved by the Medical Ethics Committee at the Affiliated People's Hospital, Fujian University of Traditional Chinese Medicine . The participants received both verbal and written information about the study and provided informed consent prior to study participation.The participants expressed their experiences on Baduanjin qigong in terms of “being sceptical,” “being suited for the elderly,” “harmonizing body and soul,” and “feeling bored.” The themes and subthemes are summarized in At the beginning of training, seven participants were sceptical about the efficacy or suitability of Baduanjin qigong in terms of health promotion. These participants talked about “doubting the efficacy” and “doubting the suitability” of Baduanjin qigong. However, with progression of the Baduanjin training program, they gradually experienced its benefit, which dispelled their doubts.Is it really effective for my health? It is composed of eight movements and appears to be simpler than other exercises. I know that Baduanjin qigong is good for my health, I learned it from TV, but I did not experience any benefit. Three participants doubted whether the Baduanjin qigong could truly benefit their physical and mental health because they thought that the exercise intensity of Baduanjin might be lower than that of other physical exercises, such as jogging, brisk walking, and square dancing. The effects of Baduanjin qigong on health promotion in elderly adults were slight; there was no effect in some cases. One participant had previously learned Baduanjin qigong by watching a TV program and practiced it intermittently for a longer period, but this participant did not experience any obvious health benefit.Maybe I cannot grasp it. I don't think it's suitable for me even though Baduanjin has only eight postures. I can never remember things because of my bad memory. Four participants did not have the confidence to learn Baduanjin qigong because of their bad memories and learning abilities. Most of them felt that it would take a long time to grasp the connotation of Baduanjin qigong, although it only consisted of eight postures.Most of the participants found that Baduanjin qigong was suitable for elderly people after the Baduanjin qigong training protocol was completed. They said that Baduanjin qigong was a “moderate amount of exercise,” was “simple and easy to learn,” and had “fewer environmental constraints.”In the past, I square-danced, I found that it was difficult for me to continue, maybe it was a problem because of my weight. I can't catch the rhythm of dance music. I felt uncomfortable and tired. I hated my fat body…However, now, I am so happy to exercise. The most mysterious part of Baduanjin qigong is that you soon get warm, but you may never get tired. Five years ago, swimming was my favourite sport. After I got stomach cancer, I often felt weak, and swimming no longer fit me. The level of Baduanjin qigong is lower than swimming, it provides a moderate amount of exercise, and I can practice Baduanjin qigong every day. There was no way to exercise with my waist pain, but I can perform Baduanjin qigong. It eased my pain. It is a gentle movement. Seventeen participants said that the level of usual exercises, such as square dancing, swimming, and softball, was too high for elderly people to adhere to practicing for 45–60 minutes each day; in addition, the movements of Baduanjin qigong were gentle and slow and provided low-moderate intensity exercise. Most of participants were able to practice Baduanjin qigong for one hour each day.It only has eight movements! It is easy for us to accept and learn. You know, our memory is poor, it's difficult for us to learn a new exercise, but Baduanjin qigong is simple, slow, and relaxing. It only took me one week to master it. All participants were surprised that it was easy for elderly adults to learn Baduanjin qigong training. They said that Baduanjin qigong was only composed of eight postures involving movement of the arms, legs, and waist and that it was simple and did not require many skills.I like to exercise, but it is always trouble for me. A sunny day and an exercise space area are necessary to practice, but it is difficult to get both at the same time. Now all problems are solved, I can perform Baduanjin qigong on my balcony without any equipment once I am free. Today is a busy day for my kids, I will perform Baduanjin qigong at home. I do it at home on rainy days. I think Baduanjin qigong is convenient, and only two things are needed: a place to stand and background music for the Baduanjin qigong. At first, I practice this exercise following the background music and a coach. Now I can do it at home without the music; the background music seems to be played in my mind. Twelve participants reported that they could practice Baduanjin qigong at home at any time because a large training space or other auxiliary apparatuses were unnecessary. They said that they could not find, thus far, another exercise that was more convenient than Baduanjin qigong.All participants described the experiential process of Baduanjin qigong harmonizing their body and mind. They talked about how Baduanjin qigong “improved physiological health” and “improved psychoemotional state” after the training.With the progression of Baduanjin qigong training, all participants became more aware of their bodies and how their bodies felt. They perceived improvements in their bodies and changed in various ways, including “relieving pain,” “enhancing digestion,” “relieving polyuria,” “strengthening immunity,” “improving the function of circulatory system,” and “improving sleep quality.”Relieving Pain. Five participants found that Baduanjin qigong could relieve pain. After they had practiced Baduanjin qigong for a few days, they found their muscular soreness, headache, or low back pain relieved gradually and even vanished later. Four participants described that they could feel the effect of qi  after Baduanjin qigong training for several weeks.The coach told us to adjust breathing to make the process of breathing smooth, then felt qi went up from the Dantian . At first, I couldn't understand, but after a few days, I really felt it. It was unbelievable; I felt that the qi went up to my chest, my nose and my head, then my stuffy nose gradually improved, and my headache was disappeared. Baduanjin qigong is very good at helping me to exercise. Because of taking care of my grandchildren, I had suffered from low back pain for quite a long time. My low back pain has been alleviated more and more after practicing Baduanjin qigong. It has not occurred again. It's really benefit to me. I had a headache and back pain before, but now I do not. When my headache or back pain attacked previously, I always used massager, but now the massager is fired. When I came here to participate in the Baduanjin qigong training, I could not raise and stretch my arms because of shoulder periarthritis. However, now look me, I can raise my arm easily without any pain. It is marvellous. Enhancing Digestion. Eight participants reported that their digestion gradually improved. They described having better appetites after participating in the Baduanjin qigong training and that these changes gradually developed, even if they were not aware of the changes taking place. Six participants also reported that their flatus and distension issues disappeared after they practiced Baduanjin qigong. Three participants had suffered from intractable constipation or flatulence for several years. Now, after practicing Baduanjin qigong for several weeks, they have regular defecation without constipation or flatulence.I benefit from Baduanjin qigong; otherwise I would not have participated for so long time. Recently my daughter told me that I eat more than I have in the past. At that time, I realized that my good appetite had returned. I used to not eat dinner because had no appetite. Now I have a good appetite. I had intractable constipation for several years. Currently, I have a bowel movement every morning after breakfast. I don't need to sit on the night stool in agony for hours. I no longer endure constipation for several days; now I have more regular bowel movements than before. I used to have uncomfortable flatulence after meal. The foods always made my stomach bloated, so I only ate a little. However, it's much better now. When I do the first or third posture of Baduanjin qigong, in which two hands reach up to the heavens to regulate San Jiao, and raise my hands on each side to adjust the spleen and stomach, I have the hiccups or fart. I feel very comfortable. Relieving Polyuria. Two participants with polyuria reported that the frequency of their polyuria or nocturia was obviously decreased after they practiced Baduanjin qigong for several weeks. Both participants were surprised at the efficacy of Baduanjin qigong on their polyuria.I went to the toilet approximately once an hour, but the volume of urine was small every time. However, I could not tolerate it if I didn't go to the toilet. After I practiced Baduanjin qigong each day for one month, the frequency of urinating decreased. Now I have control of the situation, and I have few nocturnal urination. Strengthening Immunity. Two participants reported their experiences related to immunity. One female participant said that her friends were surprised at her good complexion, but the best result was that she had more physical strength than ever, and now she could perform some housework. A participant who smoked felt that his ability to resist disease was much better after performing Baduanjin qigong for several weeks.One day, my friend stared at me in amazement. She told me that I look so good after the Baduanjin qigong. My feeling is that I am not easily fatigued, it seems that I have more physical strength. …I can travel with my family and go hiking with my friends. I have smoked for a long time, and I used to be susceptible to catching colds, but now I feel much better. Now I practice Baduanjin qigong every day, and I seem to get more strength. Improving the Circulatory System Function. Three participants reported that their blood pressure had gradually decreased after Baduanjin qigong. One participant with hypertension described the changes to his blood pressure. He has taken hypotensive drugs for 10 years. After training in Baduanjin qigong for two months, he found that his blood pressure had returned to normal.I have hypertension. I previously took hypotensive drugs every day. However, my blood pressure was gradually lowered when I performed Baduanjin qigong for two months. Now my blood pressure is almost normal, and I have stopped taking the hypotensive drugs. …I measure my blood pressure every day. In the past, my blood pressure had large fluctuations within one day. The systolic blood pressure was approximately 150 mmHg, 130 mmHg and 150 mmHg in the morning, noon and night. Recently, my systolic blood pressure stabilized to 130 mmHg, 125 mmHg and 130 mmHg in the morning, noon and night. It's more normal and stable than before. Improving Sleep Quality. Seven participants described that the quality of their sleep became better during practicing Baduanjin qigong. It was difficult for them to fall asleep before; furthermore, they experienced insomnia and dreamful sleep and were easily awoken. After they began practicing Baduanjin qigong, they noticed that their sleep had improved, their sleep time was extended, and they fell asleep quickly.In the past, although I lay down on my bed with eyes closed, the messy thinking in my brain was still working. You know, the more I want to sleep quickly, the more I cannot sleep. I was scared to sleep. …After a few weeks of training Baduanjin qigong, I can fall asleep as soon as I lied down. It's unbelievable! I never had such a good sleep quality. When I raise one hand while pressing the other hand downward (the second posture of Baduanjin qigong), my abdominal cavity was extended along with this movement. I felt comfortable. The coach told us try to melt our body into the nature. When I stood here to practice Baduanjin qigong, I image myself as a tree. I breathed slowly along with the movement spreading slowly. …It makes me relax quickly. I think it would help me in a relaxed nature sense. My body needs it. I'm in a good mood after Baduanjin qigong. My daughter always complained she had a dad with bad temper. When I met someone I didn't like, I would angry with him or her. Now I become more clam and patient, and my family relationship is more harmonious. That's all because of Baduanjin qigong. It's a positive energy! All is developing toward a good direction. I'm a post-operative gastric cancer patient, but now I feel hopeful about my life. My blood pressure is stable, and my hands feel more powerful. All participants reported that the changes in their mind-body connectedness resulted in a sense of wholeness. Sixteen participants felt comfortable and relaxed after the Baduanjin qigong. One participant shared her artistic conception of Baduanjin qigong training: she imaged herself as a tree in the nature when she was practicing Baduanjin qigong; then she could soon feel relaxed and understand the key purpose of the movements. Eight participants reported that Baduanjin qigong made them calm, pleasurable, and anxiety-free. They said that they enjoyed their exercise and seemed to obtain more vigour from it. Four participants also said that they obtained positive psychological energy and felt full hopeful about their lives.It seems boring and monotonous if I perform Baduanjin qigong for 40 minutes, I cannot concentrate my attention on one thing. One participant described that she felt that Baduanjin qigong was boring and monotonous because of its eight simple movements. She said it was difficult to maintain Baduanjin qigong training for 40 minutes every day for 12 weeks.Qi” in TCM refers to vital energy within the body. Free flow of “qi” within the meridian system is essential for good health, whereas illness would occur if qi was blocked. “Gong” means training and practice. Qigong can promote the healthy life through strengthening or balancing free flow of internal qi during consistent practice [This qualitative investigation first explored the experience and subjective feelings of elderly community adults for 12 weeks of Baduanjin qigong training. Four likely interconnected themes emerged from data analysis. The first theme: the effect of Baduanjin qigong is “being sceptical at first.” In the regimen of traditional Chinese medicine (TCM), qigong is a form of “mind-body” exercises, which simultaneously harmonize the “mind” and the “body.” “practice . Currentpractice , 23. Badpractice . However Liangshou tuo tian li sanjiao, Zuoyou kai gong si she diao, Tiaoli piwei xu dan ju, Wulao qishang xiang houqiao, Yaotou baiwei qu xinhuo, Shangshou pan zu gu shen yao, Zan quan numu zeng liqi, and Beihou qidian baibing xiao), each of them focuses on a different physical area (particular organ or system) and meridian which is a traditional Chinese medicine belief about a path through qi flows [ yin and yang by regulating qi, strengthening tendons, and improving bone strength. For instance, the third posture of Baduanjin qigong, which is called “Tiaoli piwei xu dan ju,” helps regulate the spleen and stomach through moving hands up and down over the head. The spleen (pi) and stomach (wei) are vital in the digestion and absorption processes, which are a main energy source of the human body, according to traditional Chinese medicine (TCM) [Shangshou pan zu gu shen yao” in which the practitioner uses both hands to grasp the toes along the legs. This motion can strengthen the kidneys and the waist and benefit to increase function of urinary and immune systems [ qi in the body, which is called vital energy to maintain the healthy body [ qi is closely related to psychological and mental health. The negative psychology such as angry, depressive, irritable, and restless mood may cause the stagnation of qi. On the other hand, the normal circulation of qi is also helpful to regulate the passive emotions [ qi. In this interview, participants felt their body relax, mood calm, even better their bad temper after regular Baduanjin training for several weeks. The findings indicate Baduanjin qigong's potential to improve practitioners' physical function and its benefit to promote their psychological and mental health. Consistent with the current study's findings, many quantitative studies found that regular practicing of Baduanjin qigong could promote the physical and mental health in the general population and for those with specific health challenges. For example, practicing Baduanjin could increase the capacity for physical activity and physical fitness in adults [The second theme: Baduanjin qigong can harmonize practitioner's body and soul (“harmonizing body and soul”). Baduanjin improved physical function and capacity, being reasonable in terms of the physiological effect of Baduanjin qigong. Though Baduanjin is only composed of eight separate postures  . The six systems . Therefo systems , 29. Wit systems . With rethy body . From themotions . Baduanjn adults , 33, enhn adults , relieven adults , 36, andn adults .The third theme: Baduanjin qigong is suitable for elderly populations (“being suited for the elderly”). Baduanjin is one of the more common forms of qigong, which is a relatively simple set of exercises (only eight movements) that can be completed in 20 min or less . Previou Trials [n = 20) of the total RCT participants (n = 170). A potential selective bias may have resulted from the selection of samples for this qualitative study. However, all participants were encouraged to talk in depth about their perspectives of the research topic. Furthermore, when we found parts of interviews that required modification, we visited the same participants for a second time.This study has several strengths and potential limitations. A strength of the current study was that this qualitative investigation was based on a randomized controlled trial (RCT) evaluating the complex efficacy of Baduanjin qigong intervention for which the protocol of this RCT has been published in Trials . Current Trials . A possiThe results of present study suggest that the practice of regular Baduanjin qigong training provides a number of benefits that promote the body and mind health among community elderly population. Participants reported a wide range of benefits, including promoted digestive function, circulatory system function, immunity and sleep quality, relaxed body and moods, and enhanced self-tranquility, pleasure, and symptom relief. Meanwhile they also thought that Baduanjin qigong was a suitable exercise for elderly community adults. These results indicate that Baduanjin qigong is a potentially helpful and feasible mind-body exercise that promotes the body-mind health of elderly community populations.


P < 0.001) and 30-day mortalityincreased with increasing baseline serum UA level (P < 0.001). Ina prediction analysis for 30-day mortality, the serum UA level had a cut-offconcentration of 284 µmol/L in female patients and 352 µmol/Lin male patients. Multivariate Cox proportional hazards regression analysesshowed that white blood cell counts and UA were independent prognostic factors.In conclusion, we showed that serum UA may be an independent predictor of30-day mortality in patients with paraquat poisoning.Hyperuricemia is a strong and independent predictor of all-cause mortalityin cardiovascular disease and has been found to play a role in diseases exacerbatedby oxidative stress and inflammation. This study aimed to evaluate whetherserum uric acid (UA) level is an indicator of outcome in patients with acuteparaquat poisoning. A total of 205 subjects who had attempted suicide by oralingestion of paraquat were admitted to the emergency room between January2009 and June 2014. Initial serum UA level and other laboratory parameterswere measured. A total of 66 patients died during the 30 days after admission,corresponding to a 32.2% cumulative incidence of mortality. UA levels werehigher in non-survivors than survivors ( Paraquat (PQ) is a bipyridyl, rapidly acting, nonselective herbicide widelyused in the developing world135PQ is largely secreted unchanged in urine within the first 24 hoursof ingestion. To date, the most widely accepted mechanism underlying PQ intoxicationis oxidative stress. Biotransformation of PQ in cells results in the generationof superoxide anions and subsequently other free radicals, resulting in cellularinjury such as lipid peroxidation and mitochondrial dysfunction, triggeringan inflammatory response891113Paraquat poisoning is characterized by multiple organ function failure,mainly involving the lung, kidney, heart, liver, and nervous systemThis study was a retrospective observational cohort study of patients presentingto the emergency room (ER) of The First Affiliated Hospital, College of Medicine,Zhejiang University between January 2009 and June 2014. We enrolled 221 patientswith an oral intake of paraquat from 2 to 30 mL. Patients who met thefollowing criteria were excluded: those with a history of gout (n = 8), diabetesmellitus (n = 3), hypertension (n = 2), renal failure (n = 2), and malignancy(n = 1). The remaining 205 patients  were included in thepresent study. Because UA concentrations differ significantly by gender, patientswere categorized into gender-specific tertiles based on their UA level: tertile1, UA < 329 µmol/L for men and UA < 237 µmol/Lfor women; tertile 2, 329–431 µmol/L for men and UA 237–334 µmol/Lfor women; tertile 3, UA > 431 µmol/L for men and UA >334 µmol/L for women. Informed consent was obtained from allthe subjects.2, and PaO2.All biochemical analyses were conducted using a Hitachi 7600 Clinical Analyzer, Sysmex CA-7000 System , andSysmex XE-2100 Automated Analyzer (Sysmex) using standard methods.A peripheral venous blood sample (6 mL) was collected from eachpatient within the first 24 hours after admission to the ER. Bloodsamples were used to analyze the hematological index and biochemical values.Laboratory parameters measured included: white blood cells (WBC), platelets,hemoglobin, red blood cell distribution width (RDW), neutrophil-lymphocyteratio (NLR), prothrombin time, total protein, albumin, alanine aminotransferase(ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatinekinase (CK), creatinine, potassium, pH, PaCOP value <0.05 in the univariate analysis were included in the multivariate analysis.All statistical tests were two-tailed. P < 0.05 was considered significantlydifferent.Statistical analyses were performed using SPSS, version 16 . Data are presented as the mean ± standard deviation whendata were found to be normally distributed or as the median if the distributionwas skewed. The differences among multiple groups or between two groups wereassessed using a one-way analysis of variance (ANOVA) and the Kruskal–WallisH test or Mann–Whitney U test, if appropriate. Differences by genderand in 30-day mortality among the groups were compared using a Chi-squaredtest. The area under the ROC curve was used to discriminate UA levels withrespect to 30-day mortality. Univariate and multivariate Cox regression analysesto determine predictors of 30-day mortality were presented as hazard ratioswith a 95% confidence interval. Variables that showed a This study was approved by the ethics committee of The First AffiliatedHospital, College of Medicine, Zhejiang University and was conducted in accordancewith the Declaration of Helsinki.2 graduallydecreased.We divided patients with PQ poisoning into three groups  according to the tertileof their serum UA levels. As shown in P < 0.05), LDH , and NLR ] and markers of multi-organdamage . Serum UA levels were also negatively correlated with parametersof arterial blood gases [pH  andPaCO2 ] and potassium.Serum UA levels were significantly and positively correlated with inflammatoryindexes [WBC .As shown in Sixty-six patients died during the first 30 days after admission to theER, corresponding to a 32.2% cumulative incidence of mortality. To get a deeperunderstanding of the relationship between UA level and PQ poisoning, the cumulative30-day mortality of patients was calculated by dividing the number of fatalitiesby the number of subjects in each UA tertile .The 30-P <0.001) (P <0.001). The sensitivity was 79.2% and the specificity was 60.5%.The cut-off point for UA to predict 30-day mortality in female patientswas 284 µmol/L and the area under the receiver operating characteristic(ROC) curve was 0.752  . When thAs shown in Various studies have shown that PQ primarily exerts its toxic effects throughthe redox cycle, which produces oxygen free radicals, leading to oxidativedamage and eventual cell death2122262729131432These results provide novel evidence for a strong association between UAand the mechanism of PQ toxicity. Several mechanisms could explain the significantrelationships between serum UA and mortality as a result of PQ poisoning.The association between hyperuricemia and increased risk of metabolic syndromeand cardiovascular disease has been evaluated, and hyperuricemia has beenshown to be a strong and independent predictor of all-cause mortality in community-basedstudies34363940This study has some limitations. First, we can only propose a role forUA in the etiology of PQ poisoning based on a snapshot of the circulatingUA state. This observational study was unable to definitively comment on causalityor the temporal association between high serum UA and PQ poisoning. The secondlimitation is that blood PQ levels were not assessed and PQ poisoning caseswere included on the basis of a history of oral PQ ingestion. Although, theprognostic value of plasma PQ has been previously documented in subjects withacute PQ poisoning42In summary, our results demonstrated that measurement of serum UA may bea simple and practical index for assessing the outcome of PQ poisoning. Basedon this finding, interventions that aim to decrease UA levels may have beneficialeffects in treating patients with acute PQ intoxication. Nevertheless, furtherstudies verifying the precise role of UA are needed to eventually guide developmentof clinical intervention strategies for PQ poisoning.Y.C. designed the experiments. Y.J.B., G.C.L. and J.P.W. performed theexperiments. J.W.Z. and Y.Z. wrote the majority of the manuscript text. Allauthors reviewed the final manuscript.


RANKL expression from osteoblasts, causing an increase in osteoclast mediated bone resorption. While the mechanisms involved varies slightly between tumor types, many tumors display an increase in Hedgehog signaling components that lead to increased tumor growth, therapy failure, and metastasis. The work of multiple laboratories has detailed Hh signaling in several tumor types and revealed that tumor establishment in bone can be controlled by both canonical and non-canonical Hh signaling in a cell type specific manner. This review will explore the role of Hh signaling in the modulation of tumor induced bone disease, and will shed insight into possible therapeutic interventions for blocking Hh signaling in these tumors.Despite significant progress in cancer treatments, tumor induced bone disease continues to cause significant morbidities. While tumors show distinct mutations and clinical characteristics, they behave similarly once they establish in bone. Tumors can metastasize to bone from distant sites , directly invade into bone (head and neck) or originate from the bone  where they cause pain, fractures, hypercalcemia, and ultimately, poor prognoses and outcomes. Tumors in bone secrete factors (interleukins and parathyroid hormone-related protein) that induce  Hedgehog (Hh) signaling is a complex signaling pathway that was initially identified as a developmental signaling pathway. Since its original discovery in 1980, its role in disease has been better defined. Aberrant Hh signaling has been demonstrated by numerous groups to contribute to multiple cancer types both through contributing to changes in tumor growth directly or altering the tumor microenvironment. Because of its role in multiple tumor types, Hh signaling is an attractive pathway for both basic research and drug development to identify new targets and new therapeutic approaches for inhibiting tumor growth and metastasis.Drosophila melanogaster, is an evolutionary conserved signaling pathway that is now known to have additional important roles in tissue homeostasis as well as tumorigenesis [drosophila embryos gave rise to the complex body structure of adult fruit flies. In the 1970s, Christiane Nüsslein-Volhard and Eric Wieschaus made ground breaking progress in understanding drosophila tissue patterning and polarity, which they were later awarded for with the Nobel Bell Prize in physiology or medicine. Utilizing embryonic lethal screening techniques, the pair introduced random mutations into the genome of fruit flies using ethyl methanosulfonate (EMS) and identified 15 loci important for segment number and polarity in drosophila larvae [Hh signaling, first identified by its developmental role in igenesis ,2,3,4. Ia larvae ,7,8,9,10Additionally, in context specific settings, Hh signaling has been found to function in tandem with other important developmental signaling pathways such as Wnt, BMP and TGFβ signaling ,11,12. CAs an evolutionarily conserved signaling pathway, Hh signaling works similarly in vertebrates, as outlined in PTHrP) in periarticular chondrocytes [PTHrP is secreted and diffuses along the growth plate, which increases chondrocyte proliferation in the growth plate region, and induces them to deposits large amounts of collagenous extra-cellular matrix (ECM), extending the length of the developing bone. Beyond this region, levels of PTHrP drop and chondrocytes no longer proliferate but instead undergo hypertrophy and then apoptosis [In vertebrates, both growth and ossification of the skeletal system is achieved via Hh signaling. Early in development, cellular patterning of the limb bud is controlled predominantly by Shh. Acting as a classical morphogen; secretion of Shh forms a spatial and temporal gradient which controls cellular differentiation, polarization, and proliferation . In contdrocytes ,29,30. Ppoptosis . FollowiLoss of Ihh during bone development has been shown to have detrimental effects on bone elongation and ossification . One groRANKL) on their cell surface which binds to RANK on pre-osteoclasts and facilitates their differentiation. As these cells differentiate, they fuse into large multinucleated cells. Differentiated osteoclasts secrete metalloprotein enzymes that are tartate resistant, and are known as tartate resistant acid phosphatase (TRAP) positive cells [RANKL, known as a decoy receptor because it binds free RANKL and therefore blocks RANK/RANKL binding, thus preventing further osteoclastogenesis. This coupling of osteoblast activity to osteoclast activity allows for balanced levels of bone remodeling, where bone is resorbed at about the same rate that it is made [Post-pubescent skeletons undergo constant, albeit low levels of remodeling. The two main cell types that maintain bone homeostasis are osteoblasts and osteoclasts. Osteoblasts are differentiated bone forming cells derived from mesenchymal stem cells, while osteoclasts are differentiated bone resorbing cells derived from monocytes. These cell populations have directly opposing functions that regulate one another in a tightly controlled feedback loop . Osteoblve cells . Other hve cells . Importa is made . PTHrP expression which upregulates RANKL, increasing osteoclastogenesis [PTHrP, which induces expression and secretion of RANKL. RANKL is required for osteoclastogenesis. Mature osteoclasts cause an increase in osteoclast mediated resorption of old and/or damaged bone, which is required to facilitate healing and normal bone turnover. Defects in Hh signaling have been shown to play an important role in bone remodeling. Evidence for this includes the finding that Ptch1 deficient patients show increased bone mass due to increased osteoblast differentiation . Conversogenesis . This fiogenesis ,48,49. APTCH1 gene [PTCH1, the murine skeletal abnormalities mirror those of humans [PTCH1 [PTCH1 allele non-functional. This supports the finding that sequenced BCC tumors often show PTCH1 deletions, as well as the clinical observations of BCC being most commonly diagnosed on the face, arms, back and chest; areas commonly exposed to sunlight [PTCH1 have an increased risk for other tumors as well [PTCH1, it only facilitates tumor development if the function of the other allele is disrupted. However, sporadic BCC and medulloblastoma (MB) frequently occur in patients with both PTCH1 alleles because discrete cellular alterations cause Hh signaling to become constitutively active. This often occurs genetically through mutations that directly inactivate Ptch or activate Smo, but can also occur indirectly if mutations affecting downstream regulatory proteins cause an upregulation of Hh target genes [SUFU, GLI1, and GLI2 in tumors, but not in normal surrounding stroma [Hedgehog signaling has been implicated in several stages of tumorigenesis . The firCH1 gene ,57. Thisf humans . In addis [PTCH1 . It is tsunlight ,61. Pati as well . Keratocet genes ,64,65,66g stroma ,68.GLI1 is named glioma-associated oncogene homolog1 for its association with glioblastoma multiforme (GBM) [There are several other tumors types that show dysregulation in Hh signaling. For example, me (GBM) . Althougme (GBM) ,71,72. Sme (GBM) ,74,75. Ame (GBM) ,75,76. TGLI1 expression, which is important for invasion and cell renewal [In breast and prostate cancer, activated Hh signaling is correlated with recurrence, metastasis, and ultimately lower overall survival ,80,81,82 renewal . In brea renewal . Additio renewal . The sti renewal . Thus inHedgehog pathway activation is also observed in prostate cancer, where activation can be detected in both the tumor and the stroma ,97,98. HPTHrP), which increases RANKL production in osteoblasts, stimulating osteoclastogenesis. Additional signaling factors such as interleukins 6/8/11, and TNF-α, also increase osteoclastogenesis in a similar manner, but have not been shown to be directly dependent on Hh signaling [Advanced stage tumors, as well as aggressive tumors have an increased propensity to metastasize to distant sites of the body. Along with the lung and the liver, the bone is a common site of metastasis, where tumors in the bone disrupt normal bone remodeling and cause TIBD . In patiignaling ,111,112.PTHrP, which controls bone destruction [PTHrP expression and bone destruction are significantly reduced [In breast cancer metastases to bone, this process is mediated by Hh signaling. Using bone metastatic MDA-MB-231 breast cancer cells, we have shown that TGFβ stimulation increases expression of Gli2 and truction . In an e reduced . In this reduced . A simil reduced .PTHrP [PTCH1/2, activation of Smo or a combination of both. Additionally, using the Hh inhibitor, triparanol, in a xenograph mouse model, the authors observed decreased proliferation, cellularity, and tumor size [Not surprisingly, primary bone tumors such as chondrosarcoma and osteosarcoma also dysregulate bone remodeling. Chondrosarcoma tumors originate from cartilage and are thought to arise due to activating mutations in Hh signaling . StudiesPTHrP ,118,119.mor size . in vivo model of bone destruction [Aberrant Hh signaling inducing TIBD is also observed in head and neck tumors. Oral squamous cell carcinoma (OSCC), which accounts for almost 90% of all head and neck tumors show activated Hh signaling as measured by immunohistochemical staining of Hh proteins in patient biopsies . Levels truction . Here tuin vivo revealed that the drug had low oral bioavailability, a short elimination half-life, and serious adverse effects due to toxicity [Due to its role in tumorigenesis, Hh signaling inhibitors are a promising potential therapy to inhibit tumor growth and disease, and, thus, have been tested as anti-tumor agents extensively ,126,127.toxicity . Howevertoxicity ,147,148.toxicity ,151,152.toxicity . Even sotoxicity ,157,158.toxicity . Importatoxicity .Thus, the field has now transitioned to developing Hh inhibitors that target Hh signaling downstream of Smo. Gli inhibition has been proposed as the ultimate target, as it is the effector molecule for Hh signaling. Inhibitors of Hh signaling that indirectly affect Gli are also being investigated. Gli inhibition would enable downregulation of Hh signaling in non-canonical mechanisms of Hh signaling activation, such as PI3K activation. Inhibitors that directly and indirectly inhibit Gli have been identified, and are described in in vitro and in pre-clinical studies, it remains unclear if any are potential candidates for clinical trials. The Gli antagonists, GANT58 and GANT61, were the first small molecule inhibitors determined to inhibit Hh signaling at the level of Gli protein . RecentlFour Hh pathway inhibitors HPI-1-4) have been identified by Hyman and colleagues . These c have beeCurrently, there have been several murine studies exploring the efficacy of a variety of Gli inhibitors in different tumor models ,178,179.Due to its role in tumor progression, metastasis, and treatment failure in many different tumor types, Hh signaling remains an appealing target for the development of therapeutics. Despite its promise in pre-clinical and early clinical studies, Smo inhibition alone is no longer considered an effective treatment for the inhibition of cancer growth, due to patient relapse and resistance. Because of this, combination therapies are currently being tested to evaluate efficacy in several different tumor types ,186,187.


The Tax oncoprotein promotes T-cell transformation, in part via constitutive activation of the NF-κB transcription factor; however, the underlying mechanisms remain unknown. Ubiquitination is a type of post-translational modification that occurs in a three-step enzymatic cascade mediated by E1, E2 and E3 enzymes and regulates protein stability as well as signal transduction, protein trafficking and the DNA damage response. Emerging studies indicate that Tax hijacks the ubiquitin machinery to activate ubiquitin-dependent kinases and downstream NF-κB signaling. Tax interacts with the E2 conjugating enzyme Ubc13 and is conjugated on C-terminal lysine residues with lysine 63-linked polyubiquitin chains. Tax K63-linked polyubiquitination may serve as a platform for signaling complexes since this modification is critical for interactions with NEMO and IKK. In addition to NF-κB signaling, mono- and polyubiquitination of Tax also regulate its subcellular trafficking and stability. Here, we review recent advances in the diverse roles of ubiquitin in Tax function and how Tax usurps the ubiquitin-proteasome pathway to promote oncogenesis.Human T-cell leukemia virus type 1 (HTLV-1) is a complex retrovirus that infects CD4+ T cells and causes adult T-cell leukemia/lymphoma (ATLL) in 3%–5% of infected individuals after a long latent period. HTLV-1 Tax is a  Human T-cell leukemia virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATLL) and the neurodegenerative disorder HTLV-1 associated myelopathy/tropical spastic paraparesis. It is estimated that 20 million people worldwide are infected with HTLV-1 with 3%–5% of infected individuals developing ATLL after a prolonged latent period (40–60 years) . Th. Th92]. is-Golgi . Interesis-Golgi . In addiis-Golgi . Taken tis-Golgi . Althougis-Golgi . WhetherTax also undergoes SUMOylation at sites that overlap the known ubiquitin sites although its role in NF-κB activation remains controversial . SUMOylaTax ubiquitination can be counteracted by host DUBs that cleave polyubiquitin chains to regulate Tax stability and localization. A recent study showed that CYLD, a DUB and tumor suppressor involved in NF-κB signaling pathways, interacts with Tax and removes ubiquitin chains that results in impaired Tax interaction with NEMO . In HTLVAnother important DUB that plays a prominent role in the negative regulation of key NF-κB pathways is A20  ,106. A20Tax contains NLS and NES sequences which regulate the nucleo-cytoplasmic shuttling of Tax. A fine regulation of Tax cellular distribution is controlled by specific PTMs, leading to conformational changes and inducible protein-protein interactions. The balance between Tax ubiquitination and SUMOylation may potentially regulate Tax shuttling between different cellular compartments and activation of NF-κB . The E3 CDC20 E3 ligase complex ahead of schedule leading to proteasomal degradation of cyclin B1 and securin . B. BC-termo system , we haveo system . It remaIL-17RB gene may potentially compensate for the loss of Tax-induced NF-κB in ATLL. It is also plausible that distinct receptors may be amplified or overexpressed by other mechanisms to drive NF-κB signaling in IL-17RB-negative ATLL cases.To gain more insight into the mechanisms of HTLV-1 T-cell transformation, we recently conducted a next-generation RNA sequencing study to identify genes aberrantly expressed in T cells immortalized by HTLV-1. This effort identified the IL-25 receptor subunit IL-17RB as an aberrantly overexpressed gene in HTLV-1 immortalized T cells . Tax indcis-Golgi where Tax activates IKK [In this review, we have highlighted some of the mechanisms used by the multifunctional HTLV-1 Tax oncoprotein to hijack the cellular ubiquitin-proteasome machinery to promote aberrant signaling linked to cell survival and proliferation. Ubiquitination serves as a versatile tool that can modulate Tax function including nuclear export (mono-ubiquitination), degradation in the nuclear matrix (K48-linked polyubiquitination) or NEMO binding and IKK/NF-κB activation (K63-linked polyubiquitination). Tax hijacks the host ubiquitin machinery to promote its K63-linked polyubiquitination, likely to enhance NEMO binding and other protein-protein interactions in the ates IKK ,93. Inteates IKK . These rTax also activates host E3 ligases (TRAF6) to stabilize MCL-1 and mitigate cell death triggered by genotoxic stress agents and chemotherapy drugs such as etoposide . Convers


The human T-cell leukemia/lymphoma virus, type I (HTLV-I)-encoded Tax protein is a potent transcription factor and oncoprotein. Therefore, studies on Tax function have provided critical insights into fundamental mechanisms of transcriptional activation and clues to the process of malignant transformation. Biochemical analysis of recombinant, purified Tax has yielded significant insights into many of its diverse functions, despite the absence of potentially important post-translational modifications. We will present evidence demonstrating the potent transcriptional activation function of Tax in vitro, including dissection of the interactions between Tax and specific cellular transcription factors that led to the discovery of Tax-mediated disruptions in chromatin architecture. This presentation will also describe several important findings regarding the molecular defect in Tax M47, which carries point mutations in the activation domain that render the Tax mutant defective for HTLV transcription. Comparison of the physical and functional properties of wild-type and mutant Tax reveals similar profiles in size-exclusion chromatography (SEC) and nearly identical affinity profiles in activator/co activator complex formation; paradoxical observations given the pronounced defect in the ability of Tax M47 to activate HTLV-1 transcription. Importantly, proteolysis assays revealed that TaxM47 produces a distinct digestion pattern relative to wild-type Tax, suggesting that M47 folds into a distinct tertiary structure that is not detected in SEC analysis. Together, these findings further underscore the outstanding potential of native and recombinant Tax to probe the molecular basis of transcriptional activation in higher eukaryotes, as well as the significant potential of Tax in uncovering diverse, yet fundamental, cellular processes.


With a forward cone emitting from the strong pump laser in a thermal rubidium atomic vapor, we investigate the non-degenerate parametrically amplified four-wave mixing (PA-FWM) process with dressing effects in a three-level “double-Λ” configuration both theoretically and experimentally. By seeding a weak probe field into the Stokes or anti-Stokes channel of the FWM, the gain processes are generated in the bright twin beams which are called conjugate and probe beams, respectively. However, the strong dressing effect of the pump beam will dramatically affect the gain factors both in the probe and conjugate channels, and can inevitably impose an influence on the quantum effects such as entangled degree and the quantum noise reduction between the two channels. We systematically investigate the intensity evolution of the dressed gain processes by manipulating the atomic density, the Rabi frequency and the frequency detuning. Such dressing effects are also visually evidenced by the observation of Autler-Townes splitting of the gain peaks. The investigation can contribute to the development of quantum information processing and quantum communications. S1/2, 5P1/2, 5P3/2, and 6S1/2) in a rubidium vapor, the “double-Λ” system can be more efficient due to the low requirement on phase matching condition (PMC). For the FWM process characterized as large nonlinearity at near resonance and reduced the excess noise2015The parametric four-wave mixing (FWM) process based on atomic coherence1245681012141517182022232426In this article, we demonstrate the parametrically amplified FWM (PA-FWM) process with dressing effects in an off-resonant “double-Λ” arrangement by establishing a forward cone85Rb atomic system, consisting of two hyperfine states F = 3 (|0〉) and F = 2 (|3〉) of the ground state 5S1/2 and an excited state 5P3/2 (|1〉), is used to generate the PA-FWM process. Two laser beams derived from a Ti:sapphire laser and an external cavity diode laser (ECDL) are coupled into the corresponding transitions as shown in D2 line transition of rubidium atoms, the strong pump beam E1  up to 500 mW is coupled into the cell by a polarizing beam splitter (PBS). The weak probe beam E2  with approximately 100 μW propagates in the same direction of E1 with an angle of 0.26° and is detected by a branch of a balanced homodyne photodiode detector. The generated conjugate signal that can establish the coherence between the two ground states |0〉 and |1〉 co-propagates withE2 symmetrically with respect to E1 and is received by the other branch of the balanced detector.A three-level “double-Λ” type E1 tuned far away from the resonance, the FWM process will occur in the “double-Λ” configuration, which can generate the Stokes field ESt and anti-Stokes field EASt  on the forward cone shown in i = Ωi − ωi is defined as the difference between the resonant transition frequency Ωi and the laser frequency ωi of Ei. The detected ESt and EASt signals  is the Rabi frequency between levels |i〉 ↔ |j〉, and μij is the dipole momentum; d20 = γ20 + iΔ1, d21 = γ21 + iΔ1, d10 = γ10 + i(Δ1 − ΔSt), ij is the nature decay rate between levels |i〉 and |j〉 and γij = (Γi + Γj)/2 is the decoherence rate between levels |i〉 and |j〉.where E2 can be viewed as being injected into the Stokes or anti-Stokes port of the FWM process, and the injection will serve as an optical parametric amplification (OPA) process (with PMC kASt = 2k1 − k2 or kSt = 2k1 − k2) assisted by the cascaded nonlinear process. With the perturbation chains rewritten as The presence of the weak probe beam P2 is amplified to produce an output probe with a intensity of gP2 and an output conjugate with a intensity of (g–1)P2. See Methods for the theoretical derivations of the gain factor g. Taking the dressing effects of E1 and E2 into consideration, the expression for the Stokes and conjugate anti-Stokes fields should be written asAs a result, the input probe beam with a intensity of E2 and E1 viewed as probe and coupling fields, respectively, the first-order density matrix Also, with Ip of the probe signal with gain and dressing effect can be expressed by F = 3 → F′ and F = 2 → F′, respectively, where I0 is the intensity of the probe field without Doppler absorption.Consequently, the intensity D2 line, we obtain the featured transmission signal and corresponding conjugate signal as shown in 2 = −3.5 GHz has a linewidth of 278.5 MHz and is caused by Raman absorption on 87Rb, F = 2 → F′. The dip at Δ2 = 4.5 GHz indicates the detuning of the pump field. The most prominent points are the Stokes and anti-Stokes gain peaks at the 85Rb ground-state hyperfine states . According to the experimental condition, there exists 6 GHz frequency gap between the Stokes and anti-Stokes signals in the probe channel. The seeding process can also generate the anti-Stokes and Stokes peaks in the conjugate channel corresponding to the Stokes and anti-Stokes in the probe channel, respectively. The Stokes (F = 3 → F′) in the probe channel and the corresponding anti-Stokes in the conjugate channel are strongly correlated, so do the anti-Stoke (F = 2 → F′) of the probe and Stokes of the conjugate.By experimentally scanning the probe detuning over 16 GHz across the 85Rb, F = 3 → F′ transition) and anti-Stokes  signal evolutions in the probe channel by varying the atomic intensity, respectively. The single Stokes signal profile in E1 into the Stokes port of the FWM process, and the gain intensity is related to the generated photon number described by Eq. Methods part. The EIA dip reveals the dressing effects when E1 and E2 beams are viewed as coupling and probe fields, respectively. The single anti-Stokes peak with a FWHM of 50.8 MHz in E2 beam, which can be attributed to the amplification process in Eq. F = 2 → F′ transition.G2 and G1 and the subsequent weakening is similarly due to the strengthening of the Doppler effect.The measured intensity of the Stokes gain by increasing the atomic density in G1. At small E1 power (P1), the gain can increase with G1, which is observed in P1 changing from 30 mW to 90 mW. However, due to the strong dressing effects caused by denominator terms P1 is getting larger. Also, the EIA dip can be deepened due to the increase of G1 as described by Eq. 1 being far away from the resonance. Consequently, the anti-Stokes gain described by Eq. G1 and the gain can also be much stronger than that in G1, which are depicted in F = 3 → F′ and F = 2 → F′ transitions with different G1, respectively. The anti-Stokes and Stokes signals that are conjugate to the two gain peaks in the probe channel are shown in 1, respectively. Since the horizontal positions of the two gain peaks on the probe signal can move with the pump detuning changing, the probe detuning values Δ2 in 1, which can also be consistent with the coordinate probe detuning shown in 2 = 0 and Δ2 = 6 GHz, respectively, due to the improved nonlinearity caused by the strong coherence in the FWM process at resonance. Similarly, the EIA and dressing effects can also be strengthened, which in turn can impose stronger suppression on the gain peaks. As a result of the interaction, the gain peak can have the almost unchanged intensity with Δ1 varying from Δ2 = −200 MHz to 200 MHz. The optimum choice of pump detuning Δ1 should be a balance between the increasing of gain and reduction of the dressing effect as well as the probe loss with Δ1 tuned close to the atomic line. By blocking the probe detection branch, the corresponding anti-Stokes and Stokes are received from the non-injected channel and demonstrated in 1>0 decreases with the increase of Δ1 with G1 ≫ G2 and G1 ≫ Gc considered, which can also interpret the height evolutions of the observed gain peaks well. The intensity difference changes between the probe and conjugate signals at different Δ1 are shown in F = 2 → F′ transition at different Δ1. The right image and left image in each picture are the probe signal and conjugate signal and visually demonstrate the signal intensity evolutions corresponding to 85Rb atomic system both theoretically and experimentally. With the parametrically FWM process established, we can obtain bright twin beams with gain by seeding a weak probe beam into the Stokes or anti-Stokes channel of the FWM process. Also, the dressing effects of the strong pump, the weak probe and the generated conjugate fields can impose influences on the probe gain and the conjugate gain. As an extension to our discussion, we present another powerful evidence for the dressing effects on the gain processes. To be specific, the observed Autler-Townes (AT) splittingSo far, we have presented the PA-FWM process with dressing effects assisted by the cascaded-nonlinearity process in a “double-Λ” 85Rb, F = 3 → F′ and 85Rb, F = 2 → F′, respectively, by varying the detuning of pump field. Similarly to 2 in 1. When the single-peak gain in 2 = 0.05 GHz and then the intensities of left and right peaks can change with Δ1, which means that the strong pump laser starts to dress the energy levels |1〉 and |2〉 to create corresponding primary dressing states1 tuned close to the dress state |2+〉, the left peak is much higher than the right one. Similarly, the right peak can be enhanced with Δ1 tuned close to |2−〉. Considering the parallel generation process for the anti-Stokes, the dressing states |1+〉 and |1−〉 can also explain the evolution from single peak to double peaks in With the interaction between the dressing state effects and gain processes systematically investigated by the intensity dependences of the dressed gain on the atomic density, the Rabi frequency and the frequency detuning, we can control the amplification factor in the FWM process by easily manipulating the corresponding parameters. Furthermore, the current study on controlling the dressed gain of the bright twin beams can have contribution to the development of entangled images and the generation of correlated photons source as well as squeezing states.E1 and the weak probe beam E2 have diameters of 0.7 mm and 0.4 mm respectively. The pump beam with vertical polarization is first coupled into the cell by use of a polarizing beam splitter (PBS) and then separated from the detection path via another PBS cube after the cell. The probe beam propagates in the same direction of E1 with an angle of 0.26° and intersects with the pump beam at the center inside the cell with natural abundance. The 10 mm long rubidium cell with Brewster’s angle is wrapped by μ-metal and heated by the heater tape. The weak probe and the generated conjugate beam are received by a balanced phtotodetector with 105 V/A transimpedance gain and ≈82% quantum efficiency.We use two light beams from a continuous-wave Ti:sapphire laser and an ECDL to couple the three-level “double-Λ” rubidium atomic system. The strong pump laser beam For the case of the FWM process without seeding, the generated twin-photon numbers of the Stokes and anti-Stokes channels that are proportional to the signal intensity can be shown asESt (EASt), and A and B (phase angles φ1 and φ2) defined in ESt and EASt, respectively. Here the amplification factors are the same for the two output ports of the amplifier without seeding. When the weak probe field is injected into the Stokes port of the FWM process, it can amplify the seeded signal in an appropriate condition. The photon numbers of the output Stokes and anti-Stokes fields in the amplification process with injection are described aswhere How to cite this article: Zhang, Z. et al. Dressed Gain from the Parametrically Amplified Four-Wave Mixing Process in an Atomic Vapor. Sci. Rep.5, 15058; doi: 10.1038/srep15058 (2015).


Allergic diseases are characterized by tissue eosinophilic and basophilic inflammation. There is substantial evidence that this particular inflammatory profile results from the migration to tissues of a common eosinophil-basophil (Eo/B) progenitor that undergoes a differentiative process regulated by local cytokines, termed in situ hemopoiesis. We therefore investigated the role and mechanisms of TSLP involvement in human Eo/B in situ hemopoiesis in relation to atopy. Also, since candidate gene and genome-wide association studies have identified ‘protective’ associations between the single nucleotide polymorphism (SNP) rs1837253 in the TSLP gene and risk for allergy, asthma and airway hyper-responsiveness, we evaluated the secretion of TSLP protein from primary nasal epithelial cells (NEC) in relation to rs1837253 genotype.Peripheral blood CD34+ cells derived from atopic and nonatopic individuals were stimulated with and/or IL-3, IL-5, or GM-CSF and assessed for Eo/B colony forming units (CFU) in both methylcellulose and nasal epithelial/CD34+ cell air-liquid interface (ALI) co-cultures, and cytokine surface receptor expression by flow cytometry. Genotyping was performed using a commercially available TaqMan® genotyping assay.∘ TSLP preferentially enhanced IL-3/TNFα-dependent Eo/B CFU from CD34+ cells;∘ Eo/B CFU and TSLPR expression were significantly increased in atopic-derived CD34+ cells post TSLP- (p<0.05) and IL-3/TSLP-stimulation (p<0.001), compared to non-atopic-derived CD34+ cells;∘ In progenitor/NEC ALI co-cultures, secreted TSLP was biologically active, and sufficient to induce the differentiation of CD34+ cells into Eo/B CFU;∘ There was decreased TSLP secretion by nasal epithelial cells obtained from heterozygous  and homozygous rs1837253 TSLP minor allele  individuals, compared to homozygous ‘wild-type’ individuals .These studies further support the concept of in situ hemopoiesis and point out a previously unrecognized, critical role for TSLP in the development of allergic upper airway inflammation and its role in Eo/B differentiation, a link between adaptive and innate immunity. Novel functional genomics of TSLP show that the rs1837253 polymorphism may be directly involved in the regulation of TSLP secretion, providing explanations for both genetic association studies and recently reported positive clinical trials targeting TSLP in allergic asthma.


Withania somnifera and Melia toosendan and the natural chemicals Withaferin A and Astaxanthin are new candidates for potent anti-pigmenting substances that avoid the risk of hypopigmentation.Few anti-pigmenting agents have been designed and developed according to their known hyperpigmentation mechanisms and corresponding intracellular signaling cascades. Most anti-pigmenting agents developed so far are mechanistically involved in the interruption of constitutional melanogenic mechanisms by which skin color is maintained at a normal and unstimulated level. Thus, owing to the difficulty of confining topical application to a specific hyperpigmented skin area, potent anti-pigmenting agents capable of attenuating the natural unstimulated pigmentation process have the risk of leading to hypopigmentation. Since intracellular signaling pathways within melanocytes do not function substantially in maintaining normal skin color and are activated only by environmental stimuli such as UV radiation, specifically down-regulating the activation of melanogenesis to the constitutive level would be an appropriate strategy to develop new potent anti-pigmenting agents with a low risk of hypopigmentation. In this article, we review the hyperpigmentation mechanisms and intracellular signaling pathways that lead to the stimulation of melanogenesis. We also discuss a screening and evaluation system to select candidates for new anti-melanogenic substances by focusing on inhibitors of endothelin-1 or stem cell factor-triggered intracellular signaling cascades. From this viewpoint, we show that extracts of the herbs  Also, the corresponding intracellular signaling cascades that lead to the stimulation of melanogenesis in human melanocytes have been well implicated. However, few anti-pigmenting agents have been designed and developed according to their known hyperpigmentation mechanisms and corresponding intracellular signaling cascades. Most anti-melanogenic agents developed so far inhibit tyrosinase activity , stimula2.c-Kit (c-KIT) respectively. In melasma [Hyperpigmentary disorders that appear especially on the face and are targeted by anti-pigmenting agents, include UVB-melanosis, solar lentigo and melasma. The hyperpigmentation mechanisms of those conditions have already been unraveled in detail, as depicted in  melasma , the sec melasma  is up-re melasma . This lein vivo, either by antibodies that block the corresponding receptor(s) or by receptor antagonists, is the most essential requirement to determine if a cytokine is an intrinsic factor involved in UVB hyperpigmentation. In this respect, EDN1 and SCF have been identified as intrinsic melanogenic cytokines on the basis of evidence that UVB-induced hyperpigmentation or solar lentigo are ameliorated or diminished under in vivo conditions by substances (M. chamomilla extract) capable of interrupting the EDN1-signaling cascade [c-KIT [ye/EE  mice which have wild-type functional MC1R [6 cells  [On the other hand, alpha-melanocyte stimulating hormone (αMSH) and adrenocorticotropic hormone (ACTH) have also been reported to be up-regulated both at the protein and mRNA levels upon treatment with UVB in human keratinocytes ,17. αMSH cascade ,20 or bye [c-KIT , respecte [c-KIT . The patnal MC1R . Those fnal MC1R . In contnal MC1R . Such a 8.62 pM)  at whichBased on the evidence that EDN1 and SCF are only the intrinsic melanogenic cytokines in hyperpigmentary disorders on the face being targeted by anti-pigmenting agents, it is strongly anticipated that substances able to interrupt the EDN1- or SCF-specific intracellular signaling pathways but do not directly inhibit tyrosinase activity, would be effective new anti-pigmenting agents. They would have the advantage that they do not affect normally pigmented skin where the intracellular signaling cascade is not activated, but would have distinct inhibitory effects on hyperpigmented areas of the skin with UVB-melanosis, solar lentigo and melasma. For such anti-pigmenting agents, there seems to be a low risk of eliciting hypopigmentation because they have no direct inhibitory effect on tyrosinase activity and because there is no activated intracellular signaling cascade in normally pigmented skin.3.2+ and activates PKC, respectively [In NHMs, the intracellular signaling mechanisms that lead to melanogenic activation by ligands such as EDN1 and SCF have been well elucidated, as depicted in ectively ,7. The aectively –26. Thatectively ,28. The c-KIT receptor, mediates the activation of its intrinsic tyrosine kinase activity via dimerization and subsequent autophosphorylation [c-KIT receptor then phosphorylates various substrates and associates with a number of different signaling molecules, including the SH2, Src homology domain (Shc) and growth factor receptor binding protein 2 (Grb2) adaptor proteins, and the guanine nucleotide exchange factor, SOS all of which lead to the conversion from Rat sarcoma-guanosine diphosphate (Ras-GDP) to Rat sarcoma-guanosine triphosphate (Ras-GTP) [c-KIT [On the other hand, SCF, which binds to the rylation . The actRas-GTP) –32. The Ras-GTP) . While bRas-GTP) ,34, to uRas-GTP) , tyrosinRas-GTP) , PMEL17 Ras-GTP) , EDNRB [Ras-GTP)  and c-KI) [c-KIT , all of c-KIT auto-phosphorylation (activation). That initiates synergistic c-KIT activation and leads to synergistic activation through the c-KIT/Shc/Raf-1/MEK/ERK/CREB/MITF linkage in NHMs [We have already found that there is a mutual interaction in intracellular signaling between EDN1 and SCF, which synergistically stimulates DNA synthesis and melanogenesis via cross-talk between EDN1-induced PKC activation and SCF-induced  in NHMs . Among s in NHMs . Since t4.Before screening for inhibitors of melanogenic intracellular signaling, we asked if specific signaling inhibitors can interrupt the EDN1- or SCF-induced activation of signaling molecules in acral lentigo malignant (ALM) melanoma cells, which respond to melanogenic ligands in a fashion similar to NHMs ,40. AmonWortmannin), the MEK inhibitor, PD98059, significantly suppressed the SCF-stimulated phosphorylation of ERK and of MITF at 5–30 min post-treatment in SCF-treated ALM melanoma cells, whereas H89 and Wortmannin did not affect the SCF-stimulated phosphorylation of ERK, MITF or CREB during that time  tested were gradually up-regulated over 10 days of EDN1 treatment, with a peak at days 7–10 (c-KIT) tested were gradually up-regulated over 10 days of SCF treatment with a peak at days 5 and 7 for MITF or at days 7–10 and 10–12 for the others [To develop a model for epidermal hyperpigmentary disorders that are mechanistically associated with EDN1 or SCF as intrinsic melanogenic cytokines, we used HEEs to examine the stimulatory effects of EDN1 or SCF on epidermal pigmentation ,40–42 F. The addays 7–10 . Thus, w6.Before associating the depigmenting effects of selected signaling inhibitors with the interruption of the EDN1 or SCF signaling cascades, we asked if a PKC, Phosphoinositide (PI) 3-kinase or MEK inhibitor can affect the EDN1 or SCF-stimulated pigmentation of HEEs ,42. DuriWortmannin affected the SCF-stimulated pigmentation of HEEs [Wortmannin did not. While there was no degeneration of the epidermal tissue in the MEK inhibitor-treated HEEs at day 14, increased melanin deposition throughout the epidermis was markedly abrogated in the MEK inhibitor (PD98059)-treated HEEs compared with the untreated control. In contrast, the PI3 inhibitor-treated HEEs had levels of melanin deposition similar to the untreated control. HPLC analysis of HEEs revealed that the MEK inhibitor significantly abrogated the increased level of eumelanin content (PTCA ng/mg tissue) compared with the untreated control. In contrast, the PI3 inhibitor Wortmannin did not affect the eumelanin content.We next determined whether a MEK inhibitor or the PI3 kinase inhibitor  of HEEs . During 7.Withania somnifera extract (WSE) as a candidate for an inhibitor of the EDN1-triggered intracellular signaling cascade [To search for inhibitors of EDN1 or SCF signaling, we used EDN1- or SCF-treated NHMs to test if the increased tyrosinase activity after 72 h is significantly abrogated by 3 h pre-incubation with a herb extract or natural chemical compared with the unstimulated basal level. When active substances were identified, lysates of NHMs cultured in the absence of the active substance(s) 72 h after SCF or EDN1 stimulation were directly incubated with the active substance(s) after which tyrosinase activity was measured to test if the substance(s) served as a direct inhibitor of tyrosinase activity. Substances with no direct inhibitory effect on tyrosinase activity were subjected to further evaluation. As a result of the screening of many kinds of herb extracts, we identified the  cascade . The res cascade .8.Withania somnifera, commonly known as Ashwagandha or Indian winter cherry, is a medicinal plant that contains Withaferin A (WFA) as a bioactive steroidal lactone. The known pharmacologic effects of WSE are associated with the modulation of immune function [function –43, cardfunction , protectfunction , antibacfunction  and antifunction . As for function , and it function ,50. The function . Furtherfunction .9.We next asked if the WSE interrupts the EDN1-induced phosphorylation of ERK, CREB and/or MITF. When the WSE was added 3 h prior to EDN1 stimulation, the EDN1-stimulated phosphorylation of ERK, CREB and MITF was significantly suppressed at 15 min post-treatment . To dete10.It has become evident that the WSE has a distinct potential to interrupt the EDN1-activated intracellular signaling cascade during which ERK/CREB phosphorylation (activation) leading to the up-regulation of MITF is distinctly attenuated by pre-incubation with the extract. Since the inhibition by a MEK or PKC inhibitor of ERK/CREB phosphorylation elicited by EDN1 stimulation attenuates the EDN1-stimulated pigmentation of HEEs, we asked if the WSE elicits an abrogating effect on the EDN1-stimulated pigmentation in HEEs. During the EDN1-induced stimulation of pigmentation, the addition of the WSE at concentrations of 5 or 10 μg/mL distinctly reduced the increase in visible pigmentation in a dose-dependent manner over those 14 days with the most marked suppression attained at 10 μg/mL . Those s11.WSE was accompanied by the similar down-regulated expression of MITF  involved in the anti-pigmenting effect of the WSE, we used RT-PCR analysis to examine its effects on the expression of melanocyte-specific genes during the EDN1-stimulated pigmentation of HEEs. A time course study demonstrated that the expression of the melanocyte-specific genes tested  was gradually up-regulated over 14 days with a peak at day 7. RT-PCR analysis of the effects of the WSE at day 7 revealed that the addition of the WSE at a concentration of 10 μg/mL significantly down-regulated the increased expression of those melanogenic genes compared with the untreated controls . To furt of MITF , which s12.WSE as an inhibitor of EDN1-triggerred intracellular signaling, as shown in Melia toosendan extract (MTE) has a distinct potential to interrupt the EDN1-triggered intracellular signaling cascade and also to abrogate the EDN-stimulated pigmentation of HEEs. The MTE fails to interrupt the SCF-triggered intracellular signaling cascade and consistently does not abrogate the SCF-stimulated pigmentation of HEEs [Withaferin A, one of the active chemicals in the WSE, was found to exhibit a distinct potential to interrupt both the EDN1- and SCF-triggered intracellular signaling cascades and consistently abrogates both the EDN1- and SCF-stimulated pigmentation of HEEs [In addition to the  of HEEs . Using t of HEEs . Further of HEEs ,52.13.Among the 4 types of herb extracts or natural chemicals discussed above, there is a consistency between the abilities to abrogate the up-regulated activity of tyrosinase and to interrupt the EDN1- or SCF-triggered intracellular signaling cascades and to abolish the EDN1- or SCF-stimulated pigmentation of HEEs. The sum of these findings suggests that inhibitors of EDN1- or SCF-triggered intracellular signaling are appropriate candidates for anti-pigmenting agents with a low risk of hypopigmentation and with excellent efficacy. Of course, when applied on the skin, anti-pigmenting agents are required to penetrate at effective concentrations into the epidermis where melanocytes are located in the bottom layers in order for them to exert their anti-melanogenic effects on melanocytes. In general, active chemicals included in herb extracts and natural chemicals tested have lower molecular weights and have lipophilic properties which facilitate their cutaneous permeability. Therefore, it is anticipated that inhibitors of SCF- or EDN1-triggered intracellular signaling cascades are appropriate anti-pigmenting agents capable of down-regulating the hyperpigmentation observed in UVB-melanosis, solar lentigo and melasma. It is desirable that substances capable of interrupting both the EDN1- and SCF-triggered intracellular signaling cascades are selected as candidates for new anti-pigmenting agents. However, in light of the synergistic stimulation in intracellular signaling between EDN1 and SCF, the interruption of either or both of those intracellular signaling cascades may abolish the synergistic cross-talk signaling, resulting in exerting a distinct anti-pigmenting effect on these hyperpigmentary disorders.