[Investigation and application of Dendrobium resources in Qinba Mountains with a discussion on industrial poverty alleviation model].

The Dendrobium species are rare and endangered medicinal plants, and it is difficult to investigate their wild resources with conventional methods because of typical epiphytic herbaceous. We explored Dendrobium resources(include culture resource) of Qinba Mountains and the boundary Mountain area in Hubei, Chongqing using the methods of literatures and field investigation, and found that the cultural base of Dendrobium were profound in Qinba Mountains region. Furthermore, its germplasm resources of Dendrobium were established for the first time in Wanzhou Luotian town. In case the advantages of local rock resources and poverty alleviation demand, we have actively carried out the cultivating mode of Dendrobium which grow on rock, and the poverty alleviation model of local characteristic Dendrobium industry were established preliminarily. Our application case can provide reference for the mining and transformation of traditional Chinese medicine resources census results.

[Analysis on ecological factors and active components content of wild Dipsacus asper in Chongqing Wulong district].

An HPLC method was developed for the determination of iridoid glycosides (loganin acid, loganin, sweroside) and saponins (asperosaponin Ⅵ) in the wild Dipsacus asper. A total of 108 samples consecutive growing 12 month were collected in 9 plots in Wulong district of Chongqing. Subsequent analysis of the content of loganin acid, loganin, sweroside and asperosaponin Ⅵ was performed by HPLC to evaluate the quality. In addition, 20 climate data provided by the world climate database (http://www.worldclim.org/) was analyzed to deduce the correlation between the growing environment factors and the active ingredient content accumulation of D. asperoides and choose the apposite growing environment for D. asper. The range of active ingredient content in wild D. asper were 0.01%-3.80%(loganin acid), 0.08%-0.62%(loganin), 0.12%-0.78%(sweroside), 0.64%-5.26%(asperosaponin Ⅵ). The highest content of these active ingredients was concentrated from February to April, with 2.64% of loganin acid, 0.36% of loganin), 0.57% of sweroside, and 3.09% of asperosaponin Ⅵ. The method used for determination of the active ingredient content in D. asper was simple and convenient with accurate result. The selection of the quadrats is scientific and reasonable and can be used for the analysis of the contents of the wild D. asper, thus provide a reference for quality evaluation of D. asper and protection of D. asper resources.

[Textual research on origin and genuine of Lilii Bulbus].

Lilii Bulbus, which comes from many medicinal plants，is a frequently-used traditional Chinese medicine，the records in previous herbal literatures of it's origin and quality were inconstant. To trace back it's sources, we conducted a systematical study on it's origin and quality by textual research and investigation in this paper,The result showed that the origins of Lilii Bulbus are mainly source from white-flowers, red-yellow-flowers and red-flowers of Lilium， L. brownii var. viridulum, which were believed authentic or good quality in all previous herbal literatures,and L. pumilum and L. concolor which belong to white-flowers,and L. lancifolium which belong to red-yellow-flowers were believed low-quality and unfit for medicinal uses, or they were listed below and often have different effect with L. brownii var. viridulum. Among them, only L. concolor does not belong to Lilii Bulbus according to Chinese Pharmacopeia (2015 edition), The mainstream varieties of Lilii Bulbus became L. lancifolium now according to our practical investigation, which were very different from previous herbal literatures. Although chemical and pharmacological studies provided a reference for L. lancifolium，we should respect the actual records of the previous herbal literatures，the research of material foundation of efficacy should be more sufficient，and provide science evidence for clinical application of different species of Lilii Bulbus.

Different expression of connexin 43 between culprit arteries and non-culprit arteries and role of angiotensin II on expression of connexin 43 in non-culprit arteries.

Introduction and objectives: The purpose of the study was to observe different expression of connexin 43 between culprit arteries and nonculprit arteries in ischemia-reperfusion model and investigate on the mechanism of nonculprit arteries lesions progression. Methods: Rabbit hyperlipidemia ischemia-reperfusion model was established, vascular smooth muscles of culprit arteries and nonculprit arteries were divided into 4 groups: ① culprit arteries control group, ② nonculprit arteries control group, ③ culprit arteries ischemia-reperfusion group, ④ nonculprit arteries ischemia-reperfusion group. Immunohistochemistry analysis of connexin 43 was performed in each group. Smooth muscle cells of nonculprit arteries were divided into 4 groups: ① normal control group. ② hyperlipidemia control group. ③ angiotensin II intervention group. ④ mitogen-activated protein kinase pathway inhibitor pretreatment plus angiotensin II intervention group. Expression of connexin 43 was analysed in each group. Results: Fluorescence immunohistochemistry analysis of connexin 43 showed there was significant difference between culprit arteries ischemia-reperfusion group and nonculprit arteries ischemia-reperfusion group (1723.52±138.64 vs 2136.15±237.82, P<0.001). Expression of connexin 43 in angiotensin II intervention group was higher than that in hyperlipidemia control group (1.79±0.31 vs 1.25±0.21, P<0.05), expression of connexin 43 in mitogen-activated protein kinase pathway inhibitor pretreatment plus angiotensin II intervention group was lower than that in angiotensin II intervention group [(0.85±0.19 vs 1.79±0.31, P<0.05), (0.99±0.13 vs 1.79±0.31, P<0.05), (0.81±0.15 vs 1.79±0.31, P<0.05) respectively]. Conclusions: Expression of connexin 43 in nonculprit arteries was higher than that in culprit arteries, it may be involved in angiotensin II--mitogen-activated protein kinase pathway.

Autophagy mediates free fatty acid effects on MDA-MB-231 cell proliferation, migration and invasion.

Epidemiological and animal studies indicate an association between high levels of dietary fat intake and an increased risk of breast cancer. The multifaceted role of autophagy in cancer has been revealed in previous years. However, the mechanism of this role remains unknown. In the present study, the two most common free fatty acids, palmitate acid (PA) and oleic acid (OA), were used to determine the effect on human breast cancer MDA-MB-231 cells, and the possible role of autophagy was investigated by detecting light chain 3 (LC3)-II/I. Bafliomycin A1 was used to detect autophagy flux. High palmitate acid condition-induced MDA-MB-231 cell death and invasion were mitigated by 3-methyladenine pretreatment or transfection with shRNA against autophagy protein 5. By contrast, high oleic acid condition induced MDA-MB-231 cell proliferation, migration and invasion were mitigated using rapamycin. The present results suggest that autophagy has an important role in the effects of PA and OA on breast cancer growth and metastasis in vitro.

[Resources and usage status of Tibetan medicinal plant "Bangga"].

The study is aimed to clarify the actual original plant, find out the usage status and the resource distribution of the Tibetan medicinal plant "Bangga". By using the way of the literatures survey, interview and investigation, it found out that the actual original plant of the Tibetan medicinal plant "Bangga" were the whole dried plant or the aerial part of Aconitum tanguticum or A. naviculare of Ranunculaceae, among which A. tanguticummainly distributed in Sichuan, Gansu, Qinghai, Tibet (Qamdo), and A. naviculare mainly distributed in Tibet. Sichuan, Gansu, Qinghai and other Tibetan areas mainly used the resources of A. tanguticum, Tibet (except the Qamdo area) mainly uses the A. naviculare, which resource was imminent in danger. Other species described in the literature were not used. It showed that the use of herbs related to their resources, it is recommended to strengthen the protection and guide the market.

[Dynamics of seed rain of Tripterygium hypoglaucum and soil seed bank].

Tripterygium hypoglaucum is an endangered species in arid areas of Xiannvshan Chongqing, China. The dynamic characteristics of seed rain and soil seed bank of T. hypoglaucum were studied in this paper．Results showed that T. hypoglaucum years of mature seeds distribution number up to October; the seed rain occurred from the last ten-day of September to in the first ten-day of November and the peak of scattered seed rain concentrated in the October．The numbers of soil seed bank at 2-5 cm soil layer,mainly concentrated in the 1.5-3.5 m range. T. hypoglaucum seeds to the wind as a force for transmission, the transmission ability is strong, but in the process of natural reproduction, full mature seed rate is low, the soil seed bank seeds seed short-lived factors these were unfavorable for the natural reproduction of T. hypoglaucum population.

[Analysis of varieties and standards of Primulaceae medicinal plants used in Tibetan medicine].

In this paper, the varieties and origin of Primulaceae plants that used in Tibetan medicine were analyzed. The results showed that there were 3 genera and 44 species (including the varieties) of Primulaceae plants were recorded in the relevant literatures. Among them, 17 varieties were recorded in Tibetan names, 24 varieties were recorded in Chinese names and 1 variety was used in both of them. In current quality criteria of standards at all levels in China country, 6 varieties were recorded in Tibetan names and 6 original plants were involved, which were 35% and 14% of them respectively. Seventeen varieties were recorded in Chinese name and 7 original plants were involved, which were 30% and 16% of them respectively. In Tibetan medicine standards and literatures, there were big differences between Tibetan names and Chinese names which were translated from Tibetan names and its original plants. There were only regulations of morphological identification and microscopic authentication, so the standards were very inadequate. Therefore, through literatures research, resources and current situation investigations, combining the research and specification of the name and original of Tibetan medicine, the level of normalization and standardization could be enhanced, the stable and controllable safety and utility in clinical medication could be ensured to promote advancement of industry technology Tibetan medicine.

[Effect of topographical factors on podophyllotoxin content in Sinopodophyllum hexandrum and study on ecological suitability].

In order to find the optimal topographical factor for regionslization, the content of cimetidine in 116 Sinopodophyllum hexandrum sample collected from Sichuan, Qinghai, Gansu, Tibet, Yunnan and Shaanxi provinces, was determined. Using mathematical statistics and geographical spatial analysis of GIS analysis, the relationship between content of podophyllotoxin and influencing factors including altitude gradient and gradient position was analyzed. It is found that the optimal altitude was 2 800 m to 3 600 m, the aspect of slope north or northeast and northwest and the slope 12 degrees to 65 degrees with a high suitability degree. Considering the artificial planting, the suitable planting area for S. hexandrum is comfirmed. The topographical factor is important for S. hexandrum regionalization, but has hardly effect on podophyllotoxin content. The results of the study provide an important scientific basis for S. hexandrum production development. But there are many factors which affect suitability index and podophyllotoxin content of S. hexandrum, it is necessary to consider other factors like climate and soil while exploitation and protection of S. hexandrum.

Wogonin suppresses osteopontin expression in adipocytes by activating PPARα.

AIM: Wogonin (5,7-dihydroxy-8-methoxyflavone), a major bioactive compound of the flavonoid family, is commonly extracted from the traditional Chinese medicine Scutellaria baicalensis and possesses antioxidant and anti-inflammatory activities and is assumed to have anti-diabetes function. Indeed, a current study has shown that it can possibly treat metabolic disorders such as those found in db/db mice. However, the underlying molecular mechanism remains largely unclear. The aim of this study was to investigate the impact of wogonin on osteopontin (OPN) expression in adipose tissue from type 1 diabetic mice and in 3T3-L1 adipocytes. METHODS: Type 1 diabetes was induced by streptozotocin (STZ) injection. 3T3-L1 preadipocytes were converted to 3T3-L1 adipocytes through treatment with insulin, dexamethasone, and 3-isobutyl-1-methylxanthine (IBMX). Western blot analysis and RT-PCR were performed to detect protein expression and mRNA levels, respectively. RESULTS: Wogonin treatment suppressed the increase in serum OPN levels and reduced OPN expression in adipose tissue from STZ-induced type 1 diabetic mice. Administration of wogonin enhanced PPARα expression and activity. Silencing of PPARα diminished the inhibitory effects of wogonin on OPN expression in 3T3-L1 adipocytes. Furthermore, the levels of c-Fos and phosphorylated c-Jun were reduced in wogonin-treated adipose tissue and 3T3-L1 adipocytes. In addition, wogonin treatment dramatically mitigated p38 MAPK phosphorylation. Pharmacological inhibition of p38 MAPK by its specific inhibitor SB203580 increased PPARα activity and decreased OPN expression. CONCLUSION: Our results suggest that wogonin downregulated OPN expression in adipocytes through the inhibition of p38 MAPK and the sequential activation of the PPARα pathway. Given the adverse effects of high OPN levels on metabolism, our results provide evidence for the potential administration of wogonin as a treatment for diabetes.

A risk score for no reflow in patients with ST-segment elevation myocardial infarction after primary percutaneous coronary intervention.

BACKGROUND: Many factors are associated with no-reflow (NRF) phenomenon in ST-segment elevation myocardial infarction (STEMI), including plasma glucose, age, and pre-percutaneous coronary intervention (PCI) thrombus score. Initial clinical assessment would benefit from accurate NRF prediction. This study aimed to develop a simple scoring system to predict the risk of NRF in patients undergoing primary PCI with STEMI. METHODS: Baseline clinical and procedural variables were used for risk score development (the training dataset, n = 912) and validation (the test dataset, n = 864). Independent predictors of NRF from the multivariable model were assigned integer weights based on their coefficients and incorporated into a risk score. The discriminant ability of the score was tested by receiver operating characteristic analysis using the test dataset. RESULTS: The final model included 7 significant variables, which were age, pain-to-PCI time, neutrophil count, admission plasma glucose level, pre-PCI thrombus score, collateral circulation, and Killip class. All these variables were then used to build a risk score in terms of the prediction of NRF. Receiver operating characteristic analysis demonstrated good risk prediction with a c statistic of 0.800 (95% confidence interval: 0.772-0.826) in the test dataset. CONCLUSIONS: In patients with STEMI treated by primary PCI, incidence of NRF phenomenon may be predicted with an acceptable accuracy based on a 7-item simplified risk score.

[Studies on main interspecific association of rare and endangered medicinal plant Sinopodophyllum hexandrum community in Kangding Zheduo mountain of Sichuan province].

Based on the 2 x 2 contingency table, by using multi-species relevance (variance ratio, VR), chi2-test, Ochiai index, Dice index, Jaccard index, t-test of v/x and F-test of Morisita, s index, the interspecific relationships and the spatial distribution pattern between 20 dominants in Kangding Zheduo Mountain of Sichuan province were studied. The results indicated that the interspecific association between dominants of Sinopodophyllum hexandrum community in this area did not show significant association, which suggested that the S. hexandrum community was in mature stage, and showed stronger independency, among total 190 pairs in 20 dominant species, 2 species pairs exhibited extremely significantly positive association, 12 species pairs showed significantly positive association, 6 species pairs exhibited significantly negative association and there were no pairs showed extremely significantly negative association. S. hexandrum in community did not show significant association, which indicates they are independent in community, the spatial distribution pattern of S. hexandrum is characterized by random distribution.

ER stress in adipocytes and insulin resistance: mechanisms and significance (Review).

Adipose tissue (AT) has numerous important functions within the body. Of particular importance is its role as an endocrine organ in the control of whole­body glucose and lipid metabolism, which is achieved by the secretion of various proteins. Among these proteins are adipokines, such as adiponectin, leptin, resistin, interleukin­6 and tumor necrosis factor­α. An imbalance in the expression of these adipokines occurs in a variety of conditions, such as obesity, and can lead to various metabolic abnormalities, including hyperglycemia and hyperlipidemia. In turn, this can contribute to insulin resistance (IR) and heart diseases. Adipose endoplasmic reticulum (ER) stress is increasingly recognized as the primary factor governing these conditions, which ultimately result in the initiation of IR or the aggravation of pre­existing IR. Studies have suggested that a number of conditions, including obesity, nutrient overload and metabolic syndromes, can initiate or enhance this process in a multi­dimensional manner. This review focuses on the mechanism by which ER stress in AT can contribute to IR.

PKC?-dependent activation of the ubiquitin proteasome system is responsible for high glucose-induced human breast cancer MCF-7 cell proliferation, migration and invasion.

Type 2 diabetes mellitus (T2DM) has contributed to advanced breast cancer development over the past decades. However, the mechanism underlying this contribution is poorly understood. In this study, we determined that high glucose enhanced proteasome activity was accompanied by enhanced proliferation, migration and invasion, as well as suppressed apoptosis, in human breast cancer MCF-7 cells. Proteasome inhibitor bortezomib (BZM) pretreatment mitigated high glucose-induced MCF-7 cell growth and invasion. Furthermore, high glucose increased protein kinase C delta (PKC?)-phosphorylation. Administration of the specific PKC? inhibitor rottlerin attenuated high glucose-stimulated cancer cell growth and invasion. In addition, PKC? inhibition by both rottlerin and PKC? shRNA significantly suppressed high glucose-induced proteasome activity. Our results suggest that PKC?-dependent ubiquitin proteasome system activation plays an important role in high glucose- induced breast cancer cell growth and metastasis.

[Genetic diversity and genetic structure of endangered wild Sinopodophyllum emodi by start codon targeted polymorphism].

OBJECTIVE: Revealed the genetic diversity level and genetic structure characteristics in Sinopodophyllum emodi, a rare and endangered species in China. METHOD: We detected the genetic polymorphism within and among six wild populations (45 individuals) by the approach of Start Codon Targeted (SCoT) Polymorphism. The associated genetic parameters were calculated by POP-GENE1.31 and the relationship was constructed based on UPGMA method. RESULT: A total of 350 bands were scored by 27 primers and 284 bands of them were polymorphic. The average polymorphic bands of each primer were 10.52. At species level, there was a high level of genetic diversity among six populations (PPB = 79.27%, N(e) = 1.332 7, H = 0.210 9 and H(sp) = 0.328 6). At population level, the genetic diversity level was low (PPB = 10.48% (4.00% -23.71%), N(e) = 1.048 7 (1.020 7-1.103 7), H = 0.029 7 (0.012 9-0.063 1), H(pop) = 0.046 2 (0.019 9-0.098 6). The Nei's coefficient of genetic differentiation was 0.841 1, which was consistent with the Shannon's coefficient of genetic differentiation (0.849 4). Two calculated methods all showed that most of the genetic variation existed among populations. The gene flow (N(m) = 0.094 4) was less among populations, indicating that the degree of genetic differentiation was higher. Genetic similarity coefficient were changed from 0.570 8 to 0.978 7. By clustering analysis, the tested populations were divided into two classes and had a tendency that the same geographical origin or material of similar habitats clustered into one group. CONCLUSION: The genetic diversity of samples of S. emodi is high,which laid a certain foundation for effective protection and improvement of germplasm resources.

Development and validation of a clinical risk score predicting the no-reflow phenomenon in patients treated with primary percutaneous coronary intervention for ST-segment elevation myocardial infarction.

OBJECTIVE: The 'no-reflow' phenomenon after a primary percutaneous coronary intervention (pPCI) in patients with acute ST-segment elevation myocardial infarction (STEMI) is a strong predictor of both short- and long-term mortality. We therefore developed and prospectively validated a risk score system in order to identify STEMI patients at high risk in terms of no-reflow after primary PCI. METHODS: The first part of our study used data from 1,615 STEMI patients who underwent primary PCI within 12 h from symptom onset. Using logistic regression, we derived a risk score to predict angiographic no-reflow using baseline clinical variables. From this score, we developed a simplified fast-track screen that can be used before reperfusion. In the second part of our study, we prospectively validated the score system using receiver-operating characteristic (ROC) curves with data from 692 STEMI patients. RESULTS: The model included six clinical items: age, neutrophil count, admission plasma glucose, ß-blocker treatment, time-to-hospital admission and Killip classes. The risk score system demonstrated a good risk prediction with a c-statistic of 0.757 (95% CI 0.732-0.781) based on ROC analysis. CONCLUSION: A simple risk score system based on clinical variables is useful to predict the risk of developing no-reflow after pPCI in patients with STEMI.

Effects of probucol on angiotensin II-induced BMP-2 expression in human umbilical vein endothelial cells.

Bone morphogenetic protein-2 (BMP-2) participates significantly in vascular development and pathophysiological processes. Angiotensin II (AngII) has been demonstrated to be critical in the initiation and progression of atherosclerosis. However, the effects of AngII on BMP-2 expression and of probucol on the AngII-induced BMP-2 expression in human umbilical vein endothelial cells (HUVECs) are unknown. The aim of our study was to investigate these effects. HUVECs were cultured and stimulated with various agents. The total superoxide dismutase (SOD) activity and the concentrations of malondialdehyde (MDA) and BMP-2 were measured by standard methods. Northern blotting was used to detect the expression of BMP-2 mRNA. The activation of NF-κB in the HUVECs was also determined. The AngII treatment significantly increased BMP-2 expression levels and activated NF-κB. These effects were suppressed by treatment with pyrrolidine dithiocarbamate (PDTC) or probucol. Furthermore, the increased levels of MDA in the conditioned medium and the decrease in the total SOD activity caused by the AngII treatment were reversed by treatment with probucol or PDTC. Probucol downregulated the AngII­induced BMP-2 expression. These effects of probucol may be mediated by the inhibition of NF-κB activation.

[Evaluation of medicinal reasonableness and resource utilization of Dida (Zang Yin Cheng) commonly used in Tibetan medicine].

OBJECTIVE: To evaluate the medicinal reasonableness and resource utilization of Dida from different species. METHOD: With common characteristic absorption peaks of HPLC fingerprints and SPSS cluster, the composition similarity of Dida from different species was evaluated. RESULT: The composition similarity of HPLC fingerprints of 33 Dida samples from 15 species and 1 variety originated from Swertia, Halenia, Gentianopsis, Lomatogonium was difference. The original species can be clustered into four groups by the relative area of 10 common characteristic peaks of HPLC fingerprints. The compositions of four different genera are quite different. CONCLUSION: Because of containing iridoids, xanthones, and triterpenes which have liver protection and cholagogue functions, all of species from Swertia, Halenia, Gentianopsis and Lomatogonium in Gentianaceae are classified as Dida in Tibetan medicine. According to the composition difference among different species, the HPLC fingerprints established for Dida from different source are an effective means to identify nd control the quality of Dida.

[Determination and quality assessment of 10 ingredients gentiopicroside and sweroside and so on in Tibetan medicine Jia Di (Swertia chirayita)].

OBJECTIVE: To establish a method for determination of 10 ingredients such as gentiopicroside, sweroside, and mangiferin in India swertia, and settle the index components and their limits. METHOD: By Welch materials AQ-C18 column, determination was conducted by the gradient elution with methanol and 0.4% formic acid as mobile phase, with column temperature 30 degrees C, flow rate at 1.0 mL x min(-1), and 254 nm as the detection wavelength. RESULT: The linear relatives of 10 ingredients were good. The method showed the high precision and good reproducibility, and recovery rates were between 97% and 103%. The ingredients of market com-modities varied greatly. CONCLUSION: This method is simple, sensitive, reproducible, and applicable to the determination of the main ingredients in India Swertia. Sweroside and mango glycosides were suggested as the index components for determination in Jia Di (Swertia chirayita), and their content limits are not less than 0.1%, 0.3%, respectively.