[Study on mechanism of inhibiting proliferation of head and neck cancer cells by citral, active ingredient of lemon essential oil].

To explore the active substances exerting anti-tumour effect in lemon essential oil and the molecular mechanism inhibiting the proliferation of head and neck cancer cells SCC15 and CAL33, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay(MTT) was utilized to identify the active component inhibiting the proliferation of head and neck cancer cells, namely citral. The IC_(50) of citral inhibiting the proliferation of head and neck cancer cells and normal cells were also determined. In addition, a 5-ethynyl-2'-deoxyuridine(EdU) staining assay was used to detect the effect of citral on the proliferation rate of head and neck cancer cells, and a colony formation assay was used to detect the effect of citral on tumor sphere formation of head and neck cancer cells in vitro. The cell cycle arrest and apoptosis induction of head and neck cancer cells by citral were evaluated by flow cytometry, and Western blot was used to detect the effect of citral on the expression levels of cell cycle-and apoptosis-related proteins in head and neck cancer cells. The findings indicated that citral could effectively inhibit the proliferation and growth of head and neck cancer cells, with anti-tumor activity, and its half inhibitory concentrations for CAL33 and SCC15 were 54.78 and 25.23 µg·mL~(-1), respectively. Furthermore, citral arrested cell cycle at G_2/M phase by down-regulating cell cycle-related proteins such as S-phase kinase associated protein 2(SKP2), C-MYC, cyclin dependent kinase 1(CDK1), and cyclin B. Moreover, citral increased the cysteinyl aspartate-specific proteinase-3(caspase-3), cysteinyl aspartate-specific proteinase-9(caspase-9), and cleaved poly ADP-ribose polymerase(PARP). It up-regulated the level of autophagy-related proteins including microtubule associated protein 1 light chain 3B(LC3B), sequestosome 1(P62/SQSTM1), autophagy effector protein Beclin1(Beclin1), and lysosome-associate membrane protein 1(LAMP1), suggesting that citral could effectively trigger cell apoptosis and cell autophagy in head and neck cancer cells. Furthermore, the dual-tagged plasmid system mCherry-GFP-LC3 was used, and it was found that citral impeded the fusion of autophagosomes and lysosomes, leading to autophagic flux blockage. Collectively, our findings reveal that the main active anti-proliferation component of lemon essential oil is citral, and this component has a significant inhibitory effect on head and neck cancer cells. Its underlying molecular mechanism is that citral induces apoptosis and autophagy by cell cycle arrest and ultimately inhibits cell proliferation.

Efficacy of Chinese Medicine Treatment Based on Syndrome Differentiation for Primary Insomnia: A Randomized Placebo Controlled Triple-Blinded Trial.

OBJECTIVE: To assess efficacy of Chinese medicine (CM) on insomnia considering characteristics of treatment based on syndrome differentiation. METHODS: A total of 116 participants aged 18 to 65 years with moderate and severe primary insomnia were randomized to the placebo (n=20) or the CM group (n=96) for a 4-week treatment and a 4-week follow-up. Three CM clinicians independently prescribed treatments for each patient based on syndromes differentiation. The primary outcome was change in total sleep time (TST) from baseline. Secondary endpoints included sleep onset latency (SOL), wake time after sleep onset (WASO), sleep efficiency, Pittsburgh Sleep Quality Index (PSQI) and CM symptoms. RESULTS: The CM group had an average 0.6 h more (95% confidence interval (CI): 0.3-0.9, P<0.001) TST and 34.1% (10.3%-58.0%, P=0.005) more patients beyond 0.5 h TST increment than that of the placebo group. PSQI was changed -3.3 (-3.8 to -2.7) in the CM group, a -2.0 (-3.2 to -0.8, P<0.001) difference from the placebo group. The CM symptom score in the CM group decreased -2.0 (-3.3 to -0.7, P=0.003) more than the placebo group. SOL and WASO changes were not significantly different between groups. The analysis of prescriptions by these clinicians revealed blood deficiency and Liver stagnation as the most common syndromes. Prescriptions for these clinicians displayed relative stability, while the herbs varied. All adverse events were mild and were not related to study treatment. CONCLUSION: CM treatment based on syndrome differentiation can increase TST and improve sleep quality of primary insomnia. It is effective and safe for primary insomnia. In future studies, the long-term efficacy validation and the exploratory of eutherapeutic clinicians' fixed herb formulas should be addressed (Registration No. NCT01613183).

Inhibition of macrophage MAPK/NF-κB pathway and Th2 axis by mangiferin ameliorates MC903-induced atopic dermatitis.

Available online Atopic dermatitis (AD) is a chronic, persistent inflammatory skin disease characterized by eczema-like lesions and itching. Although topical steroids have been reported for treating AD, they are associated with adverse effects. Thus, safer medications are needed for those who cannot tolerate these agents for long periods. Mangiferin (MAN) is a flavonoid widely found in many herbs, with significant anti-inflammatory and immunomodulatory activities. However, the potential modulatory effects and mechanisms of MAN in treating Th2 inflammation in AD are unknown. In the present study, we reported that MAN could reduce inflammatory cell infiltration and scratching at the lesion site by decreasing MC903-induced levels of Th2-type cytokines, Histamine, thymic stromal lymphopoietin, Leukotriene B4, and immunoglobulin E. The mechanism may be related to reductions in MAPK and NF-κB-associated protein phosphorylation by macrophages. The results suggested that MAN may be a promising therapeutic agent for AD.

A new xanthone isolated from Garcinia bracteata and its important effect on NO levels.

A new xanthone, allanxanthone F (1), and 10 known compounds were isolated from the ethanol extract of Garcinia bracteata. The structure of compound 1 was elucidated based on spectroscopic methods (UV, IR, HR-ESI-MS, and NMR). In addition, compounds 1-9 were assessed for their anti-inflammatory activities based on the expression of nitric oxide (NO) levels on lipopolysaccharide (LPS)-induced RAW264.7 macrophages, and compounds 1-3, 4 and 6-9 suggested potential anti-inflammatory activities.

Psoriasis comorbid with atherosclerosis meets in lipid metabolism.

Psoriasis (PSO) is a common skin disease affecting approximately 1%-3% of the population, and the incidence rate is increasing yearly. PSO is associated with a dramatically increased risk of cardiovascular disease, the most common of which is atherosclerosis (AS). In the past, inflammation was considered to be the triggering factor of the two comorbidities, but in recent years, studies have found that lipid metabolism disorders increase the probability of atherosclerosis in patients with psoriasis. In this review, we discuss epidemiological studies, clinical treatment methods, risk factors, and lipid metabolism of psoriasis and atherosclerosis comorbidities.

The contrasting responses of abundant and rare microbial community structures and co-occurrence networks to secondary forest succession in the subalpine region.

Knowledge of variations in abundant and rare soil microbial communities and interactions during secondary forest succession is lacking. Soil samples were gathered from different secondary successional stages (grassland, shrubland, and secondary forest) to study the responses of abundant and rare bacterial and fungal communities, interactions and driving factors to secondary forest succession by Illumina sequencing of the 16S and ITS rRNA genes. The results showed that the α-diversities (Shannon index) of abundant bacteria and fungi revealed no significant changes during secondary forest succession, but increased significantly for rare bacteria. The abundant and rare bacterial and fungal ß-diversities changed significantly during secondary forest succession. Network analysis showed no obvious changes in the topological properties (nodes, links, and average degree) of abundant microbial networks during secondary forest succession. In contrast, these properties of the rare microbial networks in the secondary forest were higher than those in the grassland and shrubland, indicating that rare microbial networks are more responsive to secondary forest succession than abundant microorganisms. Additionally, rare microbial networks revealed more microbial interactions and greater network complexity than abundant microbial networks due to their higher numbers of nodes and links. The keystone species differed between the abundant and rare microbial networks and consisted of 1 and 48 keystone taxa in the abundant and rare microbial networks, respectively. Soil TP was the most important influencing factor of abundant and rare bacterial communities. Successional stages and plant richness had the most important influences on abundant and rare fungal communities, respectively. C:P, SM and N:P were mainly related to abundant and rare microbial network topological properties. Our study indicates that abundant and rare microbial communities, interactions and driving factors respond differently to secondary forest succession.

Myocardial injury: where inflammation and autophagy meet.

Autophagy is a highly conserved bulk degradation mechanism that degrades damaged organelles, aged proteins and intracellular contents to maintain the homeostasis of the intracellular microenvironment. Activation of autophagy can be observed during myocardial injury, during which inflammatory responses are strongly triggered. Autophagy can inhibit the inflammatory response and regulate the inflammatory microenvironment by removing invading pathogens and damaged mitochondria. In addition, autophagy may enhance the clearance of apoptotic and necrotic cells to promote the repair of damaged tissue. In this paper, we briefly review the role of autophagy in different cell types in the inflammatory microenvironment of myocardial injury and discuss the molecular mechanism of autophagy in regulating the inflammatory response in a series of myocardial injury conditions, including myocardial ischemia, ischemia/reperfusion injury and sepsis cardiomyopathy.

[Changes in the structure and function of soil prokaryotic communities in subalpine Picea asperata plantations]. / äºšé«˜å±±ç²—æžäº‘æ‰äººå·¥æž—åœŸå£¤åŽŸæ ¸å¾®ç”Ÿç‰©ç¾¤è½ç»“æž„ä¸ŽåŠŸèƒ½å˜åŒ–.

The structural and functional characteristics of soil prokaryotic community are important for maintaining ecosystem functions. In this study, we examined the diversity and compositions, the key drivers, as well as functional characteristics of prokaryotic communities in the rhizosphere and non-rhizosphere soils of Picea asperata with different stand ages using high-throughput sequencing technique and bioinformatics methods. The results showed that ß-diversity of soil prokaryotic communities in both rhizosphere and non-rhizosphere showed significant differences among different stand ages, but no significant difference between rhizosphere and non-rhizosphere in the same stand age. In terms of community composition at the phylum level, the relative abundances of Proteobacteria and Rokubacteria showed an increasing trend with the increases of stand age, while the relative abundance of Actinobacteria showed a decreasing trend, but no significant difference was observed between 75 year-old planted forests (PF75) and natural forests (NF). The relative abundances of Firmicutes and Thaumarchaeota in the soil of the 25 year-old planted forests (PF25) were significantly higher than in other planted forests and NF. At the genus level, the relative abundances of RB41, Terrimonas and Acidibacter showed an increasing trend with the increases of stand age, and RB41 and Terrimonas in rhizosphere soil of PF75 were significantly higher than those in NF. Soil properties and vegetation characteristics jointly influenced the structure of soil prokaryotic communities, with herb layer coverage, soil pH, total phosphorus, and total nitrogen as major drivers. The functional characteristics of soil prokaryotic communities were significantly different among different stand ages. The relative abundances of functions involved in carbon and nitrogen cycle, e.g., cellulolysis and nitrification, decreased with the increases of stand age, whereas that of sulfate respiration involved in the sulfur cycle increased. We proposed that the structure and functional characteristics of soil prokaryotic communities could serve as important indicators of the development stages of P. asperata forests. In the later stages of plantation forest development, soil nutrient availability could be improved by mediating phosphorus-dissolving and nitrogen-enhancing microorganisms to maintain the stability of the plantation ecosystem.

Immunity: Psoriasis comorbid with atherosclerosis.

Psoriasis is an immune-mediated, persistent inflammatory disease with a genetic predisposition, and the involvement of multiple organs in psoriasis remains indicative of systemic disease. Atherosclerosis (AS) is a common complication of patients with severe or prolonged psoriasis. The specific pathogenesis of psoriasis is still unclear. Current studies suggest that psoriasis is a polygenic genetic disease with the interaction of multiple factors such as heredity and environment. Keratinocytes are proliferated through immune-mediated inflammatory pathway, which leads to cell activation, infiltration of dermis cells and release of inflammatory factors. Activation of inflammatory cells and pro-inflammatory factors play an important role in the progression of psoriasis and atherosclerosis. Studies have found that there is a close relationship between psoriasis and atherosclerosis, and systemic inflammation may be the common feature of psoriasis and AS. This paper attempts to explore the possibility of the relationship between psoriasis and atherosclerotic comorbidities from the aspects of potential epidemiology and immune mechanism, in order to provide some reference for the subsequent scientific research.

Patterns and Drivers of nirK-Type and nirS-Type Denitrifier Community Assembly along an Elevation Gradient.

nirK-type and nirS-type denitrifier communities mediate the conversion of nitrite to nitric oxide, which is the key step in denitrification. Results of previous studies have indicated that nirK-type and nirS-type denitrifiers may occupy different niches; however, the mechanisms and drivers of their responses to environmental changes within community assembly are poorly understood. In this study, we evaluated the distribution and assembly of nirK-type and nirS-type denitrifier communities along an elevation gradient from 1,800 to 4,100 m at Mount Gongga, China. Results showed that elevational patterns of alpha diversity in nirK-type and nirS-type denitrifier communities followed hump-backed patterns along the elevation gradient. However, nirK-type denitrifier communities formed two distinct clusters that were primarily separated by elevation, whereas nirS-type denitrifier communities formed three distinct clusters that were primarily separated by forest type along the elevation gradient. Moreover, deterministic processes were dominant in governing the assemblages of nirK-type and nirS-type denitrifiers. Soil pH was a key factor influencing the alpha and beta diversity of the nirK-type denitrifier communities, whereas plant richness was a primary variable influencing nirS-type denitrifiers. Additionally, our work revealed that soil denitrification potential was mainly explained by the variation in the beta diversity of denitrifier communities rather than the alpha diversity of denitrifier communities or denitrifier abundances over a large elevation gradient, and nirK-type denitrifiers played more important roles in soil denitrification. These results may contribute to predicting the consequences of global changes on denitrifier communities and their ecological services. IMPORTANCE Mount Gongga is the highest peak in the Hengduan Mountain region and is located at the southeastern fringe of the Tibetan Plateau, Sichuan Province, southwest China. As a transitional zone between the Tibetan Plateau and Sichuan Basin, Gongga Mountain features particularly diverse topography, geology, climate, and biodiversity and is a globally significant hot spot of biodiversity. In this contribution, we comprehensively describe the diversity and assembly of denitrifier communities along an elevation gradient on Gongga Mountain. Our findings established for the first time that the distribution patterns of beta diversity and driving factors differed between nirK-type and nirS-type denitrifier communities, and deterministic processes were dominant in shaping communities of denitrifiers. Moreover, the beta diversity of denitrifier communities rather than alpha diversity or denitrifier abundance played an important role in explaining denitrification potential, and the beta diversity of nirK-type denitrifier communities was more important than nirS-type denitrifier communities in soil denitrification. This work provides crucial insights into the spatial distribution of denitrifier communities and their ecological function and increases our understanding of the mechanisms underlying spatial distribution of community assembly along large elevation gradients.

[Diversity and network features of fungal community in the soils of planted and natural Picea asperata forests.] / äººå·¥ä¸Žå¤©ç„¶äº‘æ‰æž—åœŸå£¤çœŸèŒç¾¤è½å¤šæ ·æ€§åŠèŒç¾¤ç½‘ç»œå ³ç³»ç‰¹å¾.

The diversity and interactions of soil fungal community are the key to maintain the diversity and stability of ecosystem. In this study, we examined the structure, diversity and co-occurrence networks of fungal community in rhizosphere and non-rhizosphere soils of planted and natural Picea asperata forests using high-throughput sequencing technique and bioinformatic methods. The results showed that Inocybaceae and Sebacinaceae were dominant family in soils of planted and natural forests, respectively. At the genus level, Inocybe was dominant one in soils of planted and natural forests. There were significant differences in ß-diversity of fungal communities between rhizosphere and non-rhizosphere soils in both planted and natural forests. There were no significant correlations between environmental variables and the relative abundance and α-diversity of fungal communities. Herb layer coverage, soil water content, total organic carbon concentration, and plant species richness played important roles in explaining the variations of ß-diversity of fungal communities. Results of the network analysis showed that the negative correlations were dominant among soil fungal communities in natural forest, suggesting that the competition of different groups in natural forest. Moreover, there were more negative correlations in non-rhizosphere soils than in rhizosphere soils, which indicated that fungal communities in non-rhizosphere soils comprised more competitive network structure than in the rhizosphere soils. Biomarker species were identified based on differential abundance analysis. Sebacinaceae was the single shared keystone species in the fungal network which had significant differences among rhizosphere and non-rhizosphere soils of planted and natural forests. Therefore, it is suggested that the variation of differential species in the soil fungal communities between the planted and natural forest might had limited influence on the stability of the community of planted and natural forests.

Novel inorganic aromatic mixed-valent superalkali electride CaN3Ca: an alkaline-earth-based high-sensitivity multi-state nonlinear optical molecular switch.

Focusing on innovative high-performance single-pole double-throw nonlinear optical (NLO) molecular switches, two C3v configurations (1 and 3) and one D3h configuration (2) of bipyramidal CaN3Ca have been obtained by using quantum mechanical methods. Not only are 1, 2, and 3 alkaline-earth-based aromatic superalkalis, but they are also interesting electrides. The salt-like electronic structures of e-Ca2+N33-Ca2+ (1) and Ca2+N33-Ca2+e- (3) with localized redox centres are rare inorganic Robin-Day class II-type structures, and e0.5-Ca2+N33-Ca2+e0.5- (2) with a delocalized structure is a class III-type mixed-valent superalkali electride. Under a small external electric field of ±0.0110 a.u. (0.565 V Å-1), the short-distance hopping of Ca atoms in CaN3Ca from the D3h configuration with in-plane aromaticity to each C3v configuration with out-of-plane aromaticity brings about the long-range transfer of half an electron from one Ca atom to another. And, subsequently, a large dipole moment (µ0) and remarkable static first hyperpolarizability (ß0) occur. µz and ßzzz range from 0 (D3h, off form) to -12.1 or 12.1 D (C3v, on forms) and from 0 (D3h, off form) to -19 428 or 19 428 a.u. (C3v, on forms), respectively. These extremely large differences in µz and ßzzz values between the D3h and each of the C3v configurations confirm the potential of these inorganic aromatic Robin-Day-type superalkali electrides for applications in high-sensitivity multi-state nonlinear optical switches.

Icariin Acts as a Potential Agent for Preventing Cardiac Ischemia/Reperfusion Injury.

Myocardial infarction is a leading cause of mortality and morbidity worldwide. Although essential for successful recovery, myocardium reperfusion is associated with reperfusion injury. Icariin, a major flavonoid of Epimedium koreanum Nakai, has been proven to exert efficacy for improving cardiovascular function. We investigated the molecular effect and signal pathway of icariin on cardiac ischemia/reperfusion injury. In an in vivo model of infarct in rats, icariin (10 mg/kg) significantly attenuated myocardial infarct size induced by ischemia/reperfusion (I/R). From the TUNEL assay, icariin reduced the apoptotic cell induced by I/R and decreased blood indicators of creatine kinase, ischemia-modified albumin, and lactate dehydrogenase. All this effect was antagonized by the PI3K inhibitor LY294002. Meanwhile, icariin activated the PI3K/Akt/eNOS pathway. The PI3K inhibitor LY294002 suppressed icariin-mediated protective effect. These results suggest that icariin protects against myocardial ischemia reperfusion injury in rats by activating the PI3K/Akt/eNOS-dependent signal pathways and may be a useful drug for angiogenic therapy.

Low Resistance Thought Induction Sleep-regulating Technique (TIP3-2) combined with medication for primary insomnia: a randomized controlled trial.

BACKGROUND: Cognitive behavioral therapy for insomnia (CBT-I) is well-validated in the western countries. However, it has not been widely adopted or disseminated in China. One possibility is that therapeutic approaches drawn from traditional Chinese medicine (TCM) will be more widely accepted. PURPOSE: This study aimed to evaluate the efficacy of medication in combination with a therapeutic approach drawn from TCM, Low Resistance Thought Induction Sleep-regulating Technique (TIP3-2), for acute treatment of insomnia. METHOD: A randomized controlled trial was conducted. Ninety primary insomnia patients were randomly assigned to receive TIP3-2 combined with medication (n = 45) or medication only (n = 45) for 4 weeks. Medication consisted of 1-2 mg Estazolam nightly. On the basis of taking Estazolam, TIP3-2 combined with medication group was given Low Resistance Thought Induction Sleep-regulating Technique (TIP3-2) treatment twice a week. Outcomes were assessed with the Pittsburgh sleep quality index (PSQI) and polysomnography (PSG) before and after treatment. RESULTS: Both groups demonstrated significant improvements in the PSQI and polysomnography indices. The TIP3-2+ medication group demonstrated a significant difference between the two groups in PSQI total score, sleep medication use, daytime dysfunction, subjective sleep quality, as well as polysomnography indices of sleep efficiency and awakening times (P < 0.05). CONCLUSION: Among patients with primary insomnia, the addition of TIP3-2 provided benefits above and beyond the role of medication alone.

Traditional Chinese medicine herbal preparations in restless legs syndrome (RLS) treatment: a review and probable first description of RLS in 1529.

Occidental medicine has a given definition for restless legs syndrome (RLS) and knowledge of RLS pathophysiology has led to the development of its therapeutic management. RLS has no cure. Many methods have been used for its treatment, among which traditional Chinese medicine (TCM) has been considered as a new approach. However, description and management of the disease symptoms can be found in Chinese ancient medical systems. The first mention of RLS may have been as early as the third century BC described as "leg uncomfortable". Nonetheless, the lack of a complete description encompassing all four modern cardinal features of RLS makes this uncertain. On the other hand, the first description of RLS encompassing three of the four major modern criteria occurs in the ancient book of Neike Zhaiyao (Internal summary), 1529 AD just about a century and a half prior to the description of RLS by Sir Thomas Willis in England. Here, we introduce the philosophical concepts of traditional Chinese medicine and the description, classification and understanding of RLS symptoms in traditional Chinese medicine. We have conducted an in-depth review of the literature reporting one part of TCM, Chinese herbal treatment efficacy for RLS, through both English and Chinese search engines. Eighty-five studies were included in the review and more than 40 formulas (including 176 different ingredients) were found in the literature. According to the literature, Chinese herbs have been demonstrated to be safe and hold great potential to be an effective treatment modality for RLS, but the evidence is limited by the quality of these studies. Of the eighty-five studies, only nine were clinical trials with a control group and only three of them were randomized. In cases where herbal preparations were compared to Western medications for RLS, the herbal preparations appear to be superior. However, uncertainty as to whether the diagnosis of RLS was made in accord with Western norms and the use of homemade non-validated rating scales create uncertainty as to the meaning of these results. High-quality randomized and double blinded clinical trials of Chinese herbs in treating RLS will be required in the future. This review highlights aspects of Chinese herbal treatment important to guide future research and clinical practice. To our knowledge, this is the first systematic English review of the role of Chinese herbs in the treatment of RLS.

[Brief analysis on the transmission mode of traditional Chinese medicine (TCM) in ancient China].

The transmission route of TCM knowledge in ancient China primarily included doctor's transmission and non-doctor's transmission. The doctor's transmission mode included lineage-teaching system, elementary study system, transmission between schools and the clinical effects on the society etc. Non-doctor's transmission mode included literati's interests, literati's sickness, literati's health cultivation, the interests of top society, literati's recoding etc. Different transmission modes played an important role in the transmission and conservation of TCM knowledge as well as a reference to developing the transmission of present-day TCM knowledge.