Unusual N-pyridinium amino acid-type alkaloids from roots and rhizomes of Sophora tonkinensis Gagnep.

Two previously undescribed amino acid-type alkaloids with unusual N-pyridinium cation (1-2) and six known alkaloids (3-8), were isolated from the roots and rhizomes of Sophora tonkinensis Gapnea. Their structures were characterized by UV, IR, NMR, and HRESIMS spectroscopic data. The absolute configurations of compounds 1 and 2 were established through comparison of their experimental ECD spectra to the theoretical spectra of 2 calculated by TDDFT method. The plausible biosynthetic pathway of pyridinium was also proposed. Moreover, compound 4 exhibited weak XOD inhibitory activity with the inhibition rate of 65.8% at concentration of 10 µM.

Undescribed protolimonoids from the root bark of Dictamnus dasycarpus Turcz.

Nine undescribed protolimonoids, including two apotirucallane and seven tirucallane triterpenoids, as well as five known compounds, were isolated from the root bark of Dictamnus dasycarpus Turcz. Their structures were elucidated by extensive spectroscopic analysis. Compounds 4-8, with an undescribed 22,25-epoxytirucallene part, were established their absolute configuration by single-crystal X-ray diffraction of 4. Such compounds might provide evidence for the degradation of protolimonoids to limonoids, bridging an oxidative cleavage biogenetic pathway between these structurally diverse triterpenoids. None of them showed anti-inflammatory, hepatoprotective, or monoamine oxidase B inhibitory activity.

Structures and neuroprotective activities of triterpenoids from Cynomorium coccineum subsp. songaricum (Rupr.) J. Leonard.

Cynomorium coccineum subsp. songaricum (Rupr.) J. Leonard has been widely used as a Chinese herbal remedy or a functional food for treating symptoms of aging or neurodegenerative diseases. A further investigation on the finding of active constituents led to the isolation and identification of four previously undescribed triterpenoids, together with 20 known compounds. Their structures were elucidated by extensive spectroscopic analysis (IR, NMR, HRMS, and CD). Sixteen compounds showed significant neuroprotective effects against glutamate-induced or oxygen-glucose deprivation-induced SK-N-SH cell death. Our findings revealed the active constituents of C. coccineum subsp. songaricum and indicated that both oleanane-type and ursane-type triterpenes could be valuable platforms for neurodegenerative agents based on primary structure-activity relationship analysis.

Molecular evidence of herbal formula: a network-based analysis of Si-Wu decoction.

INTRODUCTION: Emerging network pharmacology (NP) combines phytochemical information with bioinformatics tools allowing herbal formulae to be illustrated holistically in the context of phytochemical basis and therapeutic mechanisms. OBJECTIVE: This study attempted to explore the holistic molecular evidence of herbal formula Si-Wu decoction (SWD) by using the method of NP. MATERIAL AND METHOD: Databases of traditional medicines combined with PubChem, SciFinder, SEA, STRING, and KEGG were employed to gather information for establishing the "compound similarity" (CS) network and the "target-(pathway)-target" (TPT) network. Gephi software was applied to visualise the networks, with further module-based and node-based network topological analysis. Moreover, the approved drugs and shortest path analysis were used to validate the TPT network. RESULTS: The CS network presented the phytochemical profile of SWD, including the major compound groups of iridoid glycosides, glycosides, phthalide lactones, phenylpropanoids, and monoterpenoids. Furthermore, the topological analysis of TPT network depicted the holistic property of SWD in interpretable neuroendocrine immunomodulation (NIM) perspective, and the node degree analysis indicated a closer connection of SWD with endocrine or metabolism system. Moreover, by combing the analysis of the CS network and TPT network, potential active ingredients could be primarily identified. CONCLUSION: The phytochemical profile and molecular target profile, which might pave the way for an understanding of SWD in modern science and provide a reference for relevant quality research and evaluation, were demonstrated by network analysis. Moreover, the methods could be further applied to discover the phytochemical or biomolecular evidence with distinct advantages in dealing with the tremendous separated information.

Yinchenhao decoction suppresses rat liver fibrosis involved in an apoptosis regulation mechanism based on network pharmacology and transcriptomic analysis.

Yinchenhao decoction (YCHD) is a classical Traditional Chinese Medicine (TCM) formula that has been widely used in the treatment of liver fibrosis caused by chronic hepatitis B and jaundice for more than 1800 years. The purpose of this study was to investigate the apoptosis regulation mechanisms of YCHD and its active components suppresses liver fibrosis. The active components and putative targets of YCHD were predicted by network pharmacology approach. Functional and pathway enrichment analysis were presented in the present study by using clusterProfiler. Further, experimental validation was done by using terminal deoxynucleotidyl transferase (TDT) dUTP nick end labelling (TUNEL) assay and western blotting in dimethylnitrosamine (DMN)-induced liver fibrosis rats, and cell proliferation assay, apoptosis assay, and western blotting in human hepatic L02 cells and LX2 cells. 45 active compounds in YCHD formula, 592 potential target proteins and 1191 liver fibrosis-related human genes were identified. Functional and pathway enrichment analysis indicated that YCHD obviously influenced TNF, PI3K-Akt signaling pathways. Further, In vivo experiment indicated that YCHD treatment not only attenuated the symptoms of liver fibrosis, but also decrease the apoptosis of hepatic parenchyma cells. Moreover, in vitro experiments showed that rhein, kaempferol and quercetin treatments remarkably decreased the protein levels of cleaved caspase-3 and increased p-ERK1/2, PI3K and Bcl-XL protein expression in TNF-α-stimulated L02 cells. On the contrary, rhein, kaempferol, aloe-emodin and quercetin inhibited the proliferation of LX2 cells and up-regulated the protein levels of Bax and cleaved caspase-8. In conclusion, 45 active components and 296 potential targets of YCHD against liver fibrosis were identified by the analysis of network pharmacology and transcriptomics combination. The mechanisms of YCHD against liver fibrosis were involved in the regulation of multiple targets, especially affecting the apoptosis-related signaling pathways.

Comparative comprehension on the anti-rheumatic Chinese herbal medicine Siegesbeckiae Herba: Combined computational predictions and experimental investigations.

ETHNOPHARMACOLOGICAL RELEVANCE: Siegesbeckiae Herba (SH) is a traditional anti-rheumatic herbal medicine in China. The SH-derived product is the first licensed traditional herbal medicinal product for the management of rheumatism-induced joint and muscle pain in United Kingdom. The authenticated plant origins listed in the official Chinese Pharmacopeia for SH include Siegesbeckia orientalis L. (SO), S. pubescens Markino (SP) and S. glabrescens Markino (SG). Although the therapeutic effects of these SH species in treating rheumatoid arthritis (RA) are similar, their difference in chemical profiles suggested their anti-rheumatisms mechanisms and effects may be different. AIM OF THE STUDY: This study was designed to comparatively comprehend the chemical and biological similarity and difference of SO, SP and SG for treating rheumatoid arthritis based on the combination of computational predictions and biological experiment investigations. MATERIALS AND METHODS: The reported compounds for SO, SP and SG were obtained from four chemical databases (SciFinder, Combined Chemical Dictionary v2009, Dictionary of Natural Products and Chinese academy of sciences Chemistry Database). The RA-relevant proteins involved in nuclear factor-kappa B (NF-κB), oxidative stress and autophagy signaling pathways were collected from the databases of Kyoto Encyclopedia of Genes and Genomes and Biocarta. The comparative comprehension of SH plants was performed using similarity analysis, molecular docking and compounds-protein network analysis. The chemical characterization of different SH extracts were qualitatively and quantitatively analyzed, and their effects on specific RA-relevant protein expressions were investigated using Western blotting analysis. RESULTS: Chemical analysis revealed that SO contains mainly sequiterpenes and pimarenoids; SP contains mainly pimarenoids, sequiterpenes, and kaurenoids; and SG contains mainly pimarenoids, flavonoids and alkaloids. Moreover, coincided with the predicted results from computational analysis, different SH species were observed to present different chemical constituents, and diverse effects on RA-relevant proteins at the biological level. CONCLUSIONS: The chemical and biological properties of SO, SP and SG were different and distinctive. The systematic comparison between these three confusing Chinese herbs provides reliable characterization profiles to clarify the pharmacological substances in SH for the precise management of rheumatism/-related diseases in clinics.

[Mechanism study on preventive and curative effects of buyang huanwu decoction in Qi deficiency and blood stasis diseases based on network analysis].

In this study, researchers adopted the network analysis method to study Buyang Huanwu decoction at three levels, namely chemical ingredients, targets and diseases, and discovered the potential effect of Buyang Huanwu decoction in cancer treatment. Besides, they analyzed the "target-target" network of Buyang Huanwu decoction based on diseases, calculated four network indexes, namely node centrality, closeness centrality, betweenness centrality and eigenvector centrality for a comprehensive evaluation on the importance and significance of each target in the network. Afterwards, key targets of Buyang Huanwu decoction were excavated to obtain two important targets--COX-2 and PPAR-gamma, which may be important targets involved in the qi deficiency and blood stasis diseases. Meanwhile, the two targets were the basis to build the core network of "chemical component-target-disease" of Buyang Huanwu decoction, which provided reference for further studies on the effect of Buyang Huanwu decoction in treating qi deficiency and blood stasis diseases. According to the study, the network analysis method was helpful to excavate potential targets Buyang Huanwu decoction in treating qi deficiency and blood stasis diseases, and could provide methodological reference for revealing the mechanism of Buyang Huanwu decoction at multiple levels, with a guiding significance for interpreting mechanisms of traditional Chinese medicinal formulae and developing new drugs.

[Influences of petroleum hydrocarbons on accumulation of cadmium and induction of metallothionein in the polychaete Perinereis aibuhitensis].

Ragworm (Polychaeta) is an ecologically important species in intertidal mudflats and estuaries and is recognized as a sentinel species for environmental monitoring. In the intertidal belt of China, the ragworm Perinereis aibuhitensis is frequently exposed to various toxicants including cadmium (Cd) and petroleum hydrocarbons (PHCs). The present study investigated the influence of PHCs on accumulation of Cd and the induction of metallothionein (MT) in P. aibuhitensis by means of chronic microcosm experiment in which the ragworms were exposed to Cd or combinations of Cd and PHCs. The accumulation of Cd in P. aibuhitensis increased significantly with the Cd exposure concentration when exposed to Cd alone. Further, the bioconcentration factors (BCFs) for Cd in P. aibuhitensis increased with the duration of exposure. The addition of PHCs in Cd exposure solutions significantly increased the accumulation of Cd in P. aibuhitensis, compared with the control. Exposure to Cd induced the expression of MT, and the expression increased with the concen- tration up to 180 mg · kg(-1) DM Cd at which point the level of induction did not increase. PHCs without Cd did not significantly induce MT in P. aibuhitensis, but the addition of PHCs with Cd did influence the induction of MT compared to Cd without PHCs. Results indicated that PHCs could modulate the expression of MT during co-exposure with Cd. The potential substances other than metals influencing the interpretation of MT expression in wild P. aibuhitensis must be recognized when MT is used as a monitoring index.

[Osteoarthritis of knee joint treated with acupuncture and moxibustion].

OBJECTIVE: To compare the clinical efficacy of osteoarthritis of knee joint treated by acupuncture and moxibustion and simple acupuncture. METHODS: Sixty-two cases were randomized into an observation group (32 cases) and a control group (30 cases). In the observation group, acupuncture and non-scarring moxibustion were applied. Acupuncture was applied on the local acupoints of knee and moxibustion was performed on Shenshu (BL 23) and Xuehai (SP 10). In the control group, only acupuncture was adopted. The clinical efficacy was observed after 2 courses of treatment. Lysholm knee joint motor function scale, visual analogue scale (VAS) and WHO quality of life (WHOQOL-BREF) were used for the assessment of scores before and after treatment and the statistical analysis of clinical efficacy. RESULTS: The total effective rate (93.8%, 30/32) in the observation group was superior to that (87.7%, 26/30) in the control group (P < 0.05). The scores in Lysholm knee joint motor function scale and VAS were improved after treatment compared with those before treatment in two groups (P < 0.05, P < 0.01). The degree of improvement in the observation group was superior to that in the control group (P < 0.05, P < 0.01). There was no statistical significance in the scores of WHOQOL-BREF before and after treatment in two groups as well as in intra-group comparison (all P > 0.05). CONCLUSION: Acupuncture and moxibustion in combination achieve the definite clinical efficacy on osteoarthritis of knee joint and this therapy is superior to simple acupuncture in the improvement of motor function of knee joint and the alleviation of pain.

[Toxic effects of petroleum hydrocarbons, copper and cadmium on polychaete Perinereis aibuhitensis Grube and on its responses in acetycholinesterase activity].

Using the indoor simulating method of pollution exposure at various concentrations, the toxic effects of various concentrations of petroleum hydrocarbons (PHCs), copper (Cu2+) and cadmium (Cd2+) on the polychaete Perinereis aibuhitensis Grube and on its acetycholinesterase (AChE) activity were examined. The results indicated that petroleum hydrocarbons, Cu2+ and Cd2+ had the high toxicity to the polychaete Perinereis aibuhitensis Grube. After 4-day and 10-day exposure to petroleum hydrocarbons, Cu2+ and Cd2+, the value of LC50 were 440 and 110 microg x L(-1) for PHCs, 1 150 and 570 microg x L(-1) for Cu2+, 5 090 and 2 500 microg x L(-1) for Cd2+. In particular, PHCs had a stronger acute toxic effect than Cu and Cd. The AChE activity was inhibited, when the animal was exposed to Cd2+ and Cu2+, but the rate of inhibition was less than 50%. In particular, the AChE activity of the animal exposed to PHCs was significantly inhibited. The maximum rate of the inhibition by PHCs reached more than 90%. Moreover, the changes in the AChE activity significantly related with the concentration of PHCs. Thus, the index in the activity of AChE can sensitively reflect the toxic effects of PHCs on Perinereis aibuhitensis Grube as an important biomarker.

[Toxic effects of petroleum hydrocarbons and copper on polychaete Nereis diversicolor and on its antioxidant enzyme systems].

Under the condition of the laboratory simulation, the toxic effects of petroleum hydrocarbons and various concentrations of copper (Cu2+) on the polychaete Nereis diversicolor and on its antioxidant enzyme defense systems were examined. The results indicate that both petroleum hydrocarbons and CU2+ have high toxicity to the polychaete. After a 3-day exposure to petroleum hydrocarbons and Cu2+, the value of LD50 was 117.5 microL x L(-1) and 864.0 microg x L(-1), respectively. The activities of peroxidase (POD) and superoxide dismutase (SOD) were influenced significantly through a 5-day exposure to single pollution of Cu2+. The activity of POD was inhibited at first and then enhanced gradually; on the contrary, the activity of SOD showed a tendency of induction firstly and then inhibition. After exposed to petroleum hydrocarbons at the concentration of the value of LD50 for 5 days, POD activity of the polychaete was not significantly induced, and the activity of SOD was lower than that of control. A 5-day exposure to the joint-pollution of petroleum hydrocarbons and Cu2+ could bring out a decrease in the activities of POD and SOD firstly and then an increase. The changes in the activity of SOD can better reflect the toxic effects of pollutants on the polychaete.

Ecological effects of crude oil residues on the functional diversity of soil microorganisms in three weed rhizospheres.

Ecological effects of crude oil residues on weed rhizospheres are still vague. The quantitative and diversity changes and metabolic responses of soil-bacterial communities in common dandelion (Taraxacum officinale), jerusalem artichoke (Silphium perfoliatum L.) and evening primrose (Acalypha australis L.) rhizospheric soils were thus examined using the method of carbon source utilization. The results indicated that there were various toxic effects of crude oil residues on the growth and reproduction of soil bacteria, but the weed rhizospheres could mitigate the toxic effects. Total heterotrophic counting colony-forming units (CFUs) in the rhizospheric soils were significantly higher than those in the non-rhizospheric soils. The culturable soil-bacterial CFUs in the jerusalem artichoke (S. perfoliatum) rhizosphere polluted with 0.50 kg/pot of crude oil residues were almost twice as much as those with 0.25 kg/pot and without the addition of crude oil residues. The addition of crude oil residues increased the difference in substrate evenness, substrate richness, and substrate diversity between non-rhizospheric and rhizospheric soils of T. officinale and A. australis, but there was no significant (p>0.05) difference in the Shannon's diversity index between non-rhizospheric and rhizospheric soils of S. perfoliatum. The rhizospheric response of weed species to crude oil residues suggested that S. perfoliatum may be a potential weed species for the effective plant-microorganism bioremediation of contaminated soils by crude oil residues.

Influences of petroleum on accumulation of copper and cadmium in the polychaete Nereis diversicolor.

Using the exposure simulation experiment, the action of petroleum affecting the accumulation of the trace metals including copper (Cu) and cadmium (Cd) in littoral polychaete Nereis diversicolor collected from the Shuangtaizi Estuary in Liaoning Province, China was examined. The results showed that there was a markedly non-linear relationship between the accumulation of Cu in worms and the experimental concentration of Cu in exposure solutions when the concentration of petroleum remained at 0, 100, and 220 microl/L, respectively. However, significantly non-linear relationship for worms exposed to Cd was observed only when the concentration of added petroleum was 0 and 220 microl/L. The accumulation of Cu in worms did not differ significantly among the three different levels of petroleum concentrations combined with various concentrations of Cu. So was the accumulation of Cd in worms (p > 0.05). However, the addition of petroleum in exposure solutions brought about an increase in the accumulation of Cu in Nereis diversicolor, in comparison with single Cu pollution. On the other hand, when the concentration of added petroleum remained at 100 microl/L, the accumulation of Cd in worms was lower than that in worms exposed to various concentrations of only cadmium. However, the worms exposed to Cd and petroleum 220 microl/L did not show obvious and identical increase in the accumulation of Cd, compared with single Cd exposure. The accumulation of both Cu and Cd in worms did not increase significantly with the increases in concentrations of Cu or Cd in exposure solutions combined with petroleum (0, 100, and 220 microl/L) under the experimental conditions. Although Nereis diversicolor is exposed to very high Cu and Cd in exposure solutions, accumulation and detoxification mechanisms are sufficient to cope with the extra metal influx in order to survive.

[Therapeutic effect of electroacupuncture combined with medicine on acute rheumatoid arthritis at active stage].

OBJECTIVE: To explore a treatment program for increasing therapeutic effect on rheumatoid arthritis at active stage. METHODS: One hundred and forty-six cases were randomly divided into treatment group (n = 74) and medicine control group (n = 72). The treatment group were treated by electroacupuncture at Quchi (LI 11), Hegu (LI 4), Yanglingquan (GB 34), etc. , combined with meloxicam, sulfasalazine and MTX. The control group treated by simple the Western medicines. Their therapeutic effects were compared. RESULTS: The effective rate was 79.73% in the treatment group and 51.39% in the control group with a significant difference between the two groups (P< 0. 05). CONCLUSION: Electroacupuncture combined with medicine has a better therapeutic effect than the simple medicine on rheumatoid arthritis at active stage.