[Fruit variation and geographical distribution of citron].

The ripe dried fruit of citron(Citrus medica) is one of the important sources of Chinese herb Citri Fructus. At the same time, it is also grown for edible and ornamental uses. There are many species and abundant genetic variation. To clarify the intraspecific variation and resource distribution of citron, this study investigated the variation in 11 citron fruits, basically covering the main species in China, including Xiaoguo citron(C. medica var. ethrog), Goucheng(C. medica var. yunnanensis), Muli citron(C.medica var. muliensis), Dehong citron(C.medica×Citrus spp.), Fuzhou citron(C.medica×C.grandis?), Mawu(C.medica×C.grandis?), Cangyuan citron, Binchuan citron, Sweet citron, Big citron, and Small citron. The natural communities of citron were proved to be mainly distributed in the southwestern and western Yunnan and southeastern Tibet of China, with Yunnan, Sichuan, Guangxi, Chongqing, Hubei, and Zhejiang identified as the main production areas. Citron has also been widely grown in India, the Mediterranean region, and the Caribbean coast countries. The field investigation revealed the large-scale intraspecific variation of citron fruits. Most of the fruits are oval-like or sphere-like in shape. The fruits are green when raw and yellow when ripe, with oil cell dots on the skin, stripe-likes running from top to bottom, and bulge at the top. Usually, in the smaller citron fruits, the pulp and juice vesicles are better developed and the central columella is tighter. By contrast, the juice vesicles and central columella in larger fruits became more vacant, with carpels visible, and the apex segregation and development of the carpels is one of the reasons for variation. These variations should be given top priority in the future variety selection and breeding, and the quality differences of different citron species and their mechanisms should be further studied. In particular, variety selection and classification management according to their medicinal or edible purposes will provide scientific and technological supports for the orderly, safe, and effective production of citron products consumed as food and medicine.

Integrated Strategy of UHPLC-Q-TOF-MS and Molecular Networking for Identification of Diterpenoids from Euphorbia fischeriana Steud. and Prediction of the Anti-Breast-Cancer Mechanism by the Network Pharmacological Method.

Breast cancer is one of the most common malignancies in women worldwide. Traditional Chinese medicine has been used as adjunctive or complementary therapy for breast cancer. Diterpenoids from Euphorbia fischeriana Steud. have been demonstrated to possess anti-breast-cancer activity. This research was aimed to systematically explore the diterpenoids from E. fischeriana and study the multiple mechanisms on breast cancer. The structures of diterpenoids were identified by the integrated strategy of UHPLC-Q-TOF-MS and molecular networking. A total of 177 diterpenoids belonging to 13 types were collected. In silico ADME analysis was performed on these compounds. It indicated that 130 of 177 diterpenoids completely adjusted to Lipinski's rule. The targets of compounds were obtained from PharmMapper. The targets of breast cancer were collected from GeneCards. Then, 197 compounds-related targets and 544 breast cancer-related targets were identified. After the intersection process, 58 overlapping targets between compounds-related targets and breast cancer-related targets were acquired. The STRING database was applied to predict the protein-protein interactions. The GO and KEGG pathway enrichment analysis were performed by using the KOBAS database. It indicated that these predicted pathways were closely related to breast cancer. The treatment effect of E. fischeriana on breast cancer might be performed through signaling pathways, such as IL-17 signaling pathway, MAPK signaling pathway, and PI3K-Akt signaling pathway. The predicted top genes such as EGFR, ESR, MAPK, SRC, CASP3, CDK2, and KDR were involved in cell proliferation, gene transcription, apoptosis, signal transduction, DNA damage and repair, tumor differentiation, metastasis, and cell cycle, which indicated that E. fischeriana might treat breast cancer comprehensively. A compounds-KEGG pathways-related targets network was built by using cytoHubba to analyze the hub compounds and targets. It concluded that E. fischeriana treated breast cancer not only by the main components but also by the microconstituents, which reflected the overall regulatory role of multicomponents treating breast cancer. To estimate the binding affinities, binding sites, and binding postures, molecular docking simulations between 177 diterpenoids and top 19 targets were carried out. The results are basically in line with expectations. In conclusion, these results can serve as references for researchers studying potential targets of diterpenoids from E. fischeriana on breast cancer in the future.

Stress Recovery of Campus Street Trees as Visual Stimuli on Graduate Students in Autumn.

Human stress recovery response to landscapes is under discussion in Chinese settings. The present study aimed to clarify the stress recovery effects of campus street trees on graduate students in autumn. A total of 150 participants (23.75 ± 1.01 years old) completed the Trier Social Stress Test (TSST) and were then randomly assigned to view one of five virtual environments, including the street trees Sophora japonica, Ginkgo biloba, Platanus acerifolia, Koelreuteria paniculata, and the indoor environment (control). Physiological responses were measured by R-R interval and electroencephalography (EEG). Psychological responses were examined through the state version of the State-Trait Anxiety Inventory (STAI-S) and the Perceived Restorativeness Scale (PRS). Results showed that R-R intervals significantly increased while viewing all street trees. Both alpha and beta brainwave activities while viewing S. japonica and G. biloba were remarkably higher than those while viewing P. acerifolia and K. paniculata. The STAI-S scores significantly decreased, and the positive PRS scores were registered after viewing street trees. We concluded that a brief virtual visual experience of campus street trees in autumn has stress recovery effects on graduate students, and the different levels of stress recovery are associated with different types of street trees.

[Serum metabolic profile involving protective effect of "Neiguan"(PC6)-electroacupuncture preconditioning in rats with myocardial ischemia reperfusion injury].

OBJECTIVE: To observe the effect of "Neiguan" (PC6)-electroacupuncture (EA) preconditioning on serum metabolites in myocardial ischemia-reperfusion injury (MIRI) rats, so as to reveal its mechanism underlying improvement of ischemic myocardium from metabonomics. METHODS: A total of 48 male SD rats were randomly divided into control, model, EA "Neiguan"(PC6) and EA "Hegu"(LI4) groups (n＝12 rats/ group). Rats of the control group were just banded on animal boards for 30 min, once daily for 7 days. The MIRI model was established by occlusion of the left anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 1 h, and rats of the model group were also banded as those in the control group. Before modeling, EA (10 Hz/50 Hz, 1 mA) was applied to bilateral "Neiguan"(PC6) and "Hegu"(LI4) for 30 min, once daily for 7 successive days. After the treatment, serum samples were collected to be analyzed by proton nuclear magnetic resonance (1H NMR) spectroscopy. The orthogonal partial least squares discriminate analysis (PLS-DA) was employed to distinguish the serum differential metabolic profile of rats in different groups and identify potential biomarkers. RESULTS: After modeling, the ECG of model group and electroacupuncture groups showed T wave towering, and there was no obvious ST segment between R wave and T wave. The T wave decreased more than 0.2 mV after reperfusion, and there was no obvious ST segment. Compared with the control group, MIRI induced significant changes of metabolites in the serum including increase of acetoacetate acid, lectic acid, creatine, glycerol and glucose, and decrease of alanine, glutamine, glycerophosphoryl choline and phosphorylcholine. In comparison with the model group, PC6-EA preconditioning induced significant changes, including an increase of glucose, and a decrease of leucine，isoleucine, valine，3-hydroxybutyric acid，lactate，acetate，acetone，acetoacetate acid，pyruvic acid，glutamine，creatine and glycerol. There is no significant difference in metabolic patterns between "Hegu" group and model group. Metabolic pathway enrichment analysis indicated that the protective effect of PC6-EA pretreatment was realized mainly by regulating pathways of glycolysis, gluconeogenesis, citric acid metabolism, pyruvate metabolism, ketone body metabolism, etc. CONCLUSION: PC6-EA pretreatment has a role in regulating gluconeogenesis, pyruvate metabolism, amino metabolism, ketone body metabolism and energy metabolism in rats with MIRI, which maybe contribute to its protective effect on ischemic myocardium, but the specific metabolic pathways and mechanisms need being studied further.

Psycho-Physiological Effects of a Peony-Viewing Program on Middle-Aged and Elderly Individuals at Different Phenological Stages.

To address the question of whether the behavior of humans to view different phenological peony flowers has various effects on their physical and mental parameters, we investigated psycho-physiological responses of 74 participants (61.3 ± 10.78 years old) to environments of pre- and post-viewing tree peonies at four stages, including the exhibition leaf stage (ELS), initial bloom stage (IBS), full bloom stage (FBS), and terminal bloom stage (TBS). Physiological factors were examined using systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), fingertip pulse (FP), blood oxygen saturation (SpO2), and psychological evaluation, which was carried out using the Profile of Mood States (POMS) and State⁻Trait Anxiety Inventory (STAI). The results indicated that the SBP, DBP, HR, and FP levels of participants were significantly reduced after viewing tree peonies, whereas no remarkable alterations in SpO2 were found. The POMS scores of anger⁻hostility (A⁻H), fatigue⁻inertia (F⁻I), tension⁻anxiety (T⁻A), confusion⁻bewilderment (C⁻B), and depression⁻dejection (D⁻D) were significantly lower, but of vigor⁻activity (V⁻A) was higher post-viewing than pre-viewing. Furthermore, participants exhibited markedly decreased anxiety levels according to the STAI. Notably, the changes in measurement indices were more pronounced at the FBS. Our studies demonstrated that a short peony-viewing program, especially at the FBS with completely opened and large tree peony flowers, would be a promising therapeutic method for improving physiological functions as well as an effective psychological relaxation strategy for middle-aged and elderly individuals.

Anti-inflammatory effect of the compounds from the flowers of Trollius chinensis.

In order to investigate the anti-inflammatory activity of flavonoids, phenolic acids, and alkaloids from the flowers of Trollius chinensis, some representative compounds, namely, orientin, 2"-O-ß-L-galactopyranosylorientin, vitexin, quercetin, isoquercetin, luteolin, veratric acid, proglobeflowery acid, trollioside, and trolline were selected to study their inhibitory effects against LPS-induced NO, IL-6, and TNF-ß release in RAW264.7 cells. At the higher concentration, both phenolic acids and flavonoids inhibited the production of NO, whereas only phenolic acids showed this effect at the lower concentration. Although trolline had stronger cytotoxicity, it exhibited a potential effect of decreasing NO production induced by LPS in the non-toxic concentration range. In addition, all tested compounds decreased the production of IL-6 and TNF-a by almost 50% at both the higher and lower concentrations. It is concluded that the anti-inflammatory activity of the phenolic acids is stronger than that of the flavonoids.

[Lignan glycosides and sucrose esters from roots of Securidaca inappendiculata].

Nine compounds, including five lignan glycosides (1-5), three sucrose esters (6-8), and one organic acid ester (9), were isolated from the ethanol extract of the roots of Securidaca inappendiculata by various chromatographic methods including silica gel, MPLC and preparative HPLC. Their structures were elucidated as acernikol-4″-O-ß-D-glucopyranoside (1), (7R, 8S)-dihydrodehydrodiconiferyl alcohol 9-O-ß-D-glucopyranoside (2), (7R, 8S)-dihydrodehydrodiconiferyl alcohol 4-O-ß-D-glucopyranoside (3), (7R, 8S)-dihydrodehydrodiconiferyl alcohol 9'-O-ß-D-glucopyranoside (4), (7R, 8S)-5-methoxydihydrodehy-drodiconiferyl alcohol 4-O-ß-D-glucopyranoside (5), 3, 6'-O-diferuloylsucrose (6), 3-O-feruloyl-6'-O-sinapoylsucrose (7), sibricose A5 (8), and mehyl ferulate (9) on the basis of 1H-, 13C-NMR and MS experiments. Compounds 1-5, 8, and 9 were isolated from the Securidaca genus for the first time. Compounds 2, 3, and 7 exhibited weak cytotoxic activities against Hela and MCF-7 cell lines.

Identification of antioxidant activity of two new aromatic ring butyrolactone derivatives from Dictamnus dasycarpus Turcz.

The present study carried out a phytochemical investigation on the root barks of Dictamnus dasycarpus Turcz, leading to the isolation and characterization of two new aromatic ring butyrolactone derivatives, dasycarpusphenol acid A (1) and dasycarpusphenol acid B (2). Their structures were elucidated by using spectroscopic techniques and HR-FAB-MS. Compounds 1 and 2 exhibited antioxidant activity, with their IC50 values being 28.95 and 41.76 mg·mL-1, respectively.

[Serum pharmacochemistry of honey-fried Stemona tuberosa Lour. by UHPLC-QTOF/MS and MetaboLynx analysis].

A high throughput UHPLC-QTOF/MS method was established to analyze and identify the constituents in rat plasma after oral administration of honey-fried Stemona tuberosa Lour. extract. Automated data analysis software MetaboLynx was developed in analysis and identification of the bioactive components and their metabolites in rat plasma following oral administration of honey-fried S. tuberosa Lour. extract. As a result, 40 compounds including 12 prototype components and 28 metabolites were characterized. The constituents absorbed into blood and the possible metabolites which were demonstrated to originate from the active fraction of honey-fried S. tuberosa Lour. were responsible for the observed efficacy. Its serum pharmacochemistry should be subjected to complete investigation to illuminate the pharmacology and active mechanism of honey-fried S. tuberosa Lour.

[Screening of active fractions with antithrombotic effect from Caragana jubata].

The antithrombotic effect of Caragana jubata (Pall.)Poir.ethanolic extract (TE)was evaluated by inferior vena cava thrombosis in rats and acute pulmonary thrombosis in mice. To search for the bioactive fractions of TE, comparison on acute pulmonary thrombosis was made between the two main fractions of TE (TE-1 and TE-2). Besides, pharmacological effects of TE, TE-1 and TE-2 on bleeding time and clotting time were also studied. Reference substances combined with UPLC/DAD-q-TOF-MS were applied to identify the main six compounds and other chemical constituents of the TE. The results showed that TE could significantly reduce the rat thrombosis weight in all doses (P<0.01) and improve the protective rate to mice in medium and high doses (P<0.05). TE-2 showed a stronger effect on protecting the mice from paralysis or death and prolonging the bleeding time and clotting time than TE-1. Chemical constituents in TE mainly include isoflavones, pterocarpans and stilbenoids. Constituents in TE-2 were mainly isoflavones and pterocarpans, while those in TE-1 were mainly stilbenoids, which could be inferred that all of these three kinds of constituents may be responsible for the antithrombotic effects of Caragana jubata.

Effect of vitamin D on clinical and biochemical parameters in polycystic ovary syndrome women: A meta-analysis.

AIM: Using a meta-analysis framework, we investigated the association between the serum level of vitamin D and the risk of polycystic ovary syndrome (PCOS) and further examined the therapeutic effect of vitamin D on the clinical features of PCOS. MATERIAL AND METHODS: Multiple databases were searched to retrieve studies. We chose clinical studies that investigated the relation between the serum level of vitamin D and the risk of PCOS or the therapeutic effect of vitamin D on PCOS. The search results were screened according to strict inclusion and exclusion criteria to select high-quality studies for inclusion. Statistical analyses were carried out using stata 12.0. RESULTS: Seventeen studies were eligible in this meta-analysis. The levels of 25-hydroxyvitamin D and the quantitative insulin-sensitivity check index in the PCOS group were remarkably lower than in the controls, whereas the homeostasis model assessment of insulin resistance in the PCOS group was markedly higher than in the controls. No statistically significant difference was observed in serum parathyroid hormone levels between the two groups. The 25-hydroxyvitamin D levels were significantly elevated after PCOS patients received vitamin D3 treatment, but serum parathyroid hormone concentration, homeostasis model assessment of insulin resistance and quantitative insulin-sensitivity check index did not show any significant changes, indicating a lack of therapeutic response. CONCLUSION: Our results suggest that the serum level of vitamin D is associated with the risk of PCOS, but the therapeutic effect of vitamin D on PCOS remains to be further explored.

Identfication of new naphthalenones from Juglans mandshurica and evalution of their anticancer activities.

Two new naphthalenone compounds were isolated from green walnut husks of Juglans mandshurica and their structures were identified as 4-butoxybutoxy-5,8-dihydroxy-3,4-dihydro-2H-naphthalen-1-one (1), 4-ethoxyethoxy-5,8-dihydroxy-3,4-dihydro-2H-naphthalen-1-one (2). Compounds 1 and 2 were named as Juglanstetralone A (1) and Juglanstetralone B (2). Compound 1 showed more significant anti-tumor activity than 2 against gastric cancer BGC-823 cells, wih the IC50 of 125.89 (g(mL(-1).

[Effect of electroacupuncture and moxibustion preconditioning on blood endothelin and creatine kinase contents and myocardial HSP 70 expression in rabbits with myocardial ischemia-reperfusion injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) and moxibustion (Moxi) preconditioning of bi- lateral "Neiguan" (PC 6) on plasma endothelin (ET) and serum creatine kinase (CK) contents and myocardial hot shock protein 70 (HSP 70) expression in myocardial ischemia-reperfusion injury (MIRI) rabbits, so as to revel their mechanisms underlying prevention of myocardial ischemia. METHODS: A total of 72 New Zealand rabbits were randomly divided into sham operation, MIRI model, EA preconditioning and Moxi preconditioning groups (n = 18/group). Each group was further divided into 0 h, 24 h and 48 h (time-point) subgroups (n=6 in each subgroup). The MIRI model was established by occlusion of the anterior descending branch of the left coronary artery for 40 min and reperfusion for 60 min. The contents of plasma ET and serum CK were detected by ELISA, and myocardial HSP 70 expression was detected by immunohistochemistry. EA and Moxi preconditioning were respectively applied to bilateral PC 6 for 20 min, once daily for 5 days. RESULTS: Following MIRI, contents of plasma ET and serum CK contents were significantly increased at 0 h, 24 h and 48 h in comparison with the sham group (P<0.01, P<0.05), while myo- cardial HSP 70 expression at the 3 time-points was moderately increased (P>0.05). Compared with the model groups, plasma ET contents at both 24 h and 48 h in the EA preconditioning group and at 48 h in the Moxi preconditioning group, CK contents at both 24 h and 48 h only in the EA preconditioning group were significantly down-regulated (P<0.01, P<0.05). Myocardial HSP 70 expression levels in the EA and Moxi preconditioning groups were considerably up-regulated at the three time-points in comparison with the model group(P<0.05, P<0.01). CONCLUSION: Acupuncture and moxibustion pretreatment may suppress MIRI-induced increase of plasma ET and serum CK and up-regulate myocardial HSP 70 protein expression in MIRI rabbits, suggesting a preventive protection action on ischemic myocardium.

[Effect of electroacupuncture intervention on spinal AMPA-receptor expression in rats with neuropathic pain].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on changes of spinal a-amino-3- ydroxy-5-methylisoxazole-4-propionate acid (AMPA) receptor (GluR 1) expression in rats with chronic constrictive injury (CCI) pain, so as to explore its mechanisms underlying improvement of neuropathic pain. METHODS: Sixty male SD rats were randomly divided into sham-operation, CCI model and EA groups (n=20). Neuropathic pain model was established by ligature of the right sciatic nerve. EA stimulation (2 Hz, 1-3 mA) was applied to "Weizhong" (BL 40) and "Huantiao" (GB 30) on the injured limb for 30 min, once a day for 7 days beginning from the 11th day on after CCI. The mechanical and thermal pain thresholds were measured before and after the CCI procedure (baseline) and after EA intervention. The AMPA receptor subunit GluR 1 protein and gene expression in L5-L 6 segments of the spinal cord was detected using Western blot (WB), immunohistochemistry and re- verse transcription (RT)-polymerase chain reaction (PCR), separately. RESULTS: As the results of mechanical and thermal pain thresholds in our past study, EA intervention could markedly raise CCl-reduced decreased pain threshold. Compared with the sham-operation group, the expression levels of spinal GluR 1 protein and mRNA in the model group were significantly increased (P<0.05, P<0.01). Following EA intervention, the expression levels of GluR 1 protein and mRNA were remarkably down-regulated in the EA group in comparison with those of the model group (all P<005). CONCLUSION: EA intervention can down-regulate CCI-induced increase of AMPA receptor GIuR 1 expression in the lumbar spinal cord in CCI rats, which may contribute to its effect in alleviating neuropathic pain.

[Effect of manual acupuncture stimulation of "Baihui" (GV 20), etc. on serum IFN-gamma and IL-4 contents in rats with chronic fatigue syndrome].

OBJECTIVE: To observe the effect of manual acupuncture stimulation of "Baihui" (GV 20), etc. on serum IFN- gamma and IL-4 contents in rats with chronic fatigue syndrome (CFS). METHODS: A total of 24 male SD rats were equally randomized into control group, model group and acupuncture group. CFS model was established by bounding and forced swimming in cold water once daily for 14 days. Rats in the acupuncture group were treated by manual acupuncture stimulation of bilateral "Zusanli" (ST 36), "Baihui" (GV 20), and "Guanyuan" (CV 4), once daily for 14 days. Serum IFN-gamma and IL-4 contents were detected by ELISA. RESULTS: Compared with the control group, the contention of serum IFN-gamma and ratio of IFN-gamma/IL-4 were significant decreased in the model group (P<0.01). While in comparison with the model group, the contention of IFN-gamma and ratio of IFN-gamma/IL-4 were obviously increased in the acupuncture group (P<0.05). No significant differences were found among the three groups in serum IL-4 levels (P>0.05). CONCLUSION: Manual acupuncture can inhibit CFS induced reduction of serum IFN-gamma level and the ratio of IFN-gamma/IL-4 in CFS rats, suggesting a favorable adjustment of acupuncture intervention for CFS by balancing the ratio of IFN-gamma/IL-4.

Two new compounds from Dictamnus dasycarpus.

Two new compounds, one new pyrrolidine alkaloid and one new eudesmane-type sesquiterpene derivative, named, respectively, as dictamnaindiol and dictamnadiol, were isolated from the EtOAc soluble fraction of the ethanolic extract from the root barks of Dictamnus dasycarpus. Their structures were elucidated as (3α,4ß)-3-(6-ethoxy-6-(4-hydroxyphenyl)methyl)-4-(4-hydroxyphenyl)-1-methyl-pyrrolidin-2-one (1) and (1R, 4S, 7S, 10R)-1,11-dihydroxyl-4,14-epoxy-5(6)-eudesmene (2) on the basis of their spectral and chemical analysis.

Three new compounds from Dictamnus dasycarpus.

Phytochemical investigation of the ethyl acetate extracts from root barks of Dictamnus dasycarpus led to the isolation of three new compounds, named as dasycarpusenester A (1), dasycarpusester B (2), dasycarpusacid (3). Their structures were elucidated as (2S)-4-(2,2-dimethyl-5-oxotetrahydrofuran-3-yl)-2-hydroxypent-3-enoic acid methyl ester (1), (2R)-4-(2,2-dimethyl-5-oxotetrahydrofuran-3-yl)-2-hydroxypent-3-anoic acid methyl ester (2), and (2S)-4-(2,2-dimethyl-5-oxotetrahydrofuran-3-yl)-2-hydroxypent-3-anoic acid (3), respectively, on the basis of modern spectroscopic methods and chemical analysis.