[Regulation of ischemic stroke by circadian rhythm and intervention by traditional Chinese medicine].

Circadian rhythm is an internal regulatory mechanism formed in organisms in response to the circadian periodicity in the environment, which modulates the pathophysiological events, occurrence and development of diseases, and the response to treatment in mammals. It significantly influences the susceptibility, injury, and recovery of ischemic stroke, and the response to therapy. Accumulating evidence indicates that circadian rhythms not only regulate the important physiological factors of ischemic stroke events, such as blood pressure and coagulation-fibrinolysis system, but also participate in the immuno-inflammatory reaction mediated by glial cells and peripheral immune cells after ischemic injury and the regulation of neurovascular unit(NVU). This article aims to link molecular, cellular, and physiological pathways in circadian biology to the clinical consequences of ischemic stroke and to illustrate the impact of circadian rhythms on ischemic stroke pathogenesis, the regulation of NVU, and the immuno-inflammatory responses. The regulation of circadian rhythm by traditional Chinese medicine is reviewed, and the research progress of traditional Chinese medicine intervention in circadian rhythm is summarized to provide a reasonable and valuable reference for the follow-up traditional Chinese medicine research and molecular mechanism research of circadian rhythm.

[Influences of municipal sludge applied in slope vegetation restoration on surface water environment]. / 城市污泥应用于边坡植被恢复对地表水环境的影响.

The application of municipal sludge in ecological restoration has a good prospect for avoiding the food chain of grain crops, but its influences on surface water environmental are unclear. The municipal sludge and construction waste were mixed with 1:1 (V/V) as growth media, which were covered over simulation coal gangue slopes. Eight native woody species were sowed in the mixed media. The plant growth and coverage, as well as conductivity, pH, the concentrations of nitrogen (N), phosphorus (P), potassium (K), heavy metal and polycyclic aromatic hydrocarbon (PAHs) of surface and underground runoff of the slopes in the growing season were investigated. The results showed that plants grew well on the mixed media. The average plant coverage reached 60%. The pH of the surface and underground runoff changed little and near to neutral. The conductivity, N, P, K, heavy metal and PAHs contents of the slope runoff were high. The N and P contents in the growing season were above the National Standards of Surface Water Quality (GB 3838-2002) V. The contents of heavy metal were the highest in July. The contents of As lied at the GB IV-V, whereas other heavy metal contents up to GB II-IV. With strong rain leaching in the summer as well as the absorption, degrading and fix effect of plant-soil system on chemical substrates, the conductivity and N, P, K, heavy metal and PAHs contents of the slope runoff significantly decreased. The contents of heavy metal in late stage of growing season arrived at GB 2-3. The contents of PAHs reduced by about 50%. The direct application of municipal sludge in ecological restoration of coal gangue slope were beneficial to plant growth. The plant-soil system might gradually decrease the harmful substance concentrations in the growth media. The negative influences on surface water environment mainly came from eutrophication of N and P. Generally, the environmental safety is manageable.

Gegen Qinlian Decoction Attenuates High-Fat Diet-Induced Steatohepatitis in Rats via Gut Microbiota.

Gut microbiota play an important role in modulating energy contribution, metabolism, and inflammation, and disruption of the microbiome population is closely associated with chronic metabolic diseases, such as nonalcoholic fatty liver disease (NAFLD). Gegen Qinlian decoction (GGQLD), a well-known traditional Chinese herbal medicine (CHM), was previously found to regulate lipid metabolism and attenuate inflammation during NAFLD pathogenesis. However, the underlying mechanism of this process, as well as how the gut microbiome is involved, remains largely unknown. In this study, we investigated the effect of varying doses of GGQLD on the total amount and distribution of gut bacteria in rats fed a high-fat diet (HFD) for 8 weeks. Our analysis indicates that Oscillibacter and Ruminococcaceae_g_unclassified are the dominant families in the HFD group. Further, HFD-dependent differences at the phylum, class, and genus levels appear to lead to dysbiosis, characterized by an increase in the Firmicutes/Bacteroidetes ratio and a dramatic increase in the Oscillibacter genus compared to the control group. Treatment with GGQLD, especially the GGQLL dose, improved these HFD-induced changes in intestinal flora, leading to increased levels of Firmicutes, Clostridia, Lactobacillus, bacilli, and Erysipelotrichales that were similar to the controls. Taken together, our data highlight the efficacy of GGQLD in treating NAFLD and support its clinical use as a treatment for NAFLD/NASH patients.

Clerosterol from vinegar-baked radix bupleuri modifies drug transport.

Vinegar-baked Radix Bupleuri (VBRB) is reportedly used to treat liver cancer when combined with traditional chemotherapy and data show that this combination may modify drug transport. We isolated clerosterol from VBRB and studied its effect on drug transporters in normal or transporter-overexpressing cells. Transporter activity was assayed using cellular substrate concentration and transporter expression with Western blot and RT-qPCR. Clerosterol decreased cisplatin uptake in BRL cells mainly through increasing Mrp2 gene expression. Clerosterol also decreased the uptake of colchicine in HEK 293 cells by increasing both Pgp and Mrp1 activity; in detail, it could increase Pgp protein but had marginal effects on Mrp1 protein and gene expression. Further study showed clerosterol increased OCT2 activity in HEK293-Pgp cells by increasing OCT2 protein and mRNA. Clerosterol could suppress Pgp overexpression but not by regulating protein and gene expression. And clerosterol had marginal effects on Mrp2 and Mrp1 activity in Mrp2- and Mrp1-overexpressing HEK293 cells. Thus, Clerosterol may be an active constituent of VBRB and may work against cancer multidrug resistance by inhibiting Pgp activity.

Intervening TNF-α via PPARγ with Gegenqinlian Decoction in Experimental Nonalcoholic Fatty Liver Disease.

This paper is to explore the effect and mechanism of Gegenqinlian decoction on experimental nonalcoholic fatty liver disease (NAFLD) in vivo and in vitro. The final aim is to make clear whether Gegenqinlian decoction would impact NAFLD through improving PPARγ to suppress inflammation and regulate lipid. The data in this research suggested that Gegenqinlian decoction is a potent way to manage NAFLD through improving PPARγ to regulate lipid and suppress inflammation.

Discovery of a small-molecule inhibitor of STAT3 by ligand-based pharmacophore screening.

STAT3 modulates the transcription of a wide variety of regulatory genes involved in cell proliferation, differentiation, migration, apoptosis, and other critical cellular functions. Constitutive activation of STAT3 has been detected in a wide spectrum of human malignancies. A pharmacophore model constructed from a training set of STAT3 inhibitors binding to the SH2 domain was used to screen an in-house database of compounds, from which azepine 1 emerged as a top candidate. Compound 1 inhibited STAT3 DNA-binding activity in vitro and attenuated STAT3-directed transcription in cellulo with comparable potency to the well-known STAT3 inhibitor S3I-201. A fluorescence polarization assay revealed that compound 1 targeted the SH2 domain of STAT3. Furthermore, compound 1 inhibited STAT3 phosphorylation in cells without affecting the total expression of STAT3. This study also validates the use of pharmacophore modeling to identify inhibitors of protein-protein interactions.

Addition of optically pure H-phosphinate to ketones: selectivity, stereochemistry and mechanism.

Aromatic methyl ketones and cyclic asymmetric ketones underwent hydrophosphorylation with P-stereogenic H-P species in the presence of potassium carbonate to produce P,C-stereogenic tertiary α-hydroxyl phosphinates in excellent yields with up to 99 : 1 dr. The diastereoselectivity was induced by a reversible conversion of less stable stereomer of product to that of a more stable one via an equilibrium, which was confirmed by aldehyde/ketone exchanging reaction. Toward the exchange, aliphatic or aldehyde carbonyl were more active than aromatic or ketone carbonyls, respectively. The stability difference between the two diastereomers was controlled by the sizes of substituents linking to phosphorus or α-carbon.

Discovery of a natural product-like iNOS inhibitor by molecular docking with potential neuroprotective effects in vivo.

In this study, we applied structure-based virtual screening techniques to identify natural product or natural product-like inhibitors of iNOS. The iNOS inhibitory activity of the hit compounds was characterized using cellular assays and an in vivo zebrafish larvae model. The natural product-like compound 1 inhibited NO production in LPS-stimulated Raw264.7 macrophages, without exerting cytotoxic effects on the cells. Significantly, compound 1 was able to reverse MPTP-induced locomotion deficiency and neurotoxicity in an in vivo zebrafish larval model. Hence, compound 1 could be considered as a scaffold for the further development of iNOS inhibitors for potential anti-inflammatory or anti-neurodegenerative applications.

Effects of Nardostachys chinensis on spontaneous ventricular arrhythmias in rats with acute myocardial infarction.

OBJECTIVES: To investigate the effects and mechanisms of Nardostachys chinensis (NC) on spontaneous ventricular arrhythmias in rats with hyper-acute myocardial infarction (AMI). METHODS: Seventy-two rats were randomly divided into the control group (n = 24), metoprolol group (n = 24), and the NC group (n = 24). Premature ventricular contractions (PVCs), ventricular tachycardias (VTs), ventricular fibrillations (VFs), and blood pressure were monitored for 4 hours after coronary artery ligation. The connexin 43 (Cx43) expression in ventricular myocardium was measured by immunohistochemistry, Western blot, and real-time RT-PCR. RESULTS: Compared with the control, metoprolol and NC decreased the VF incidence (50% vs. 4.2%, P < 0.001, and 50% vs. 12.5%, P = 0.005, respectively). There was a steady decrease in the cumulative number of PVCs and VTs within 4 hours from ligating in 3 groups. Compared with the control, metoprolol and NC reduced the cumulative number of VTs and PVCs. Compared with control, metoprolol and NC decreased the infarct size of the left ventricular tissue (55.98% ± 6.20% vs. 39.13% ± 4.53%, P < 0.001, and 55.98% ± 6.20% vs. 42.39% ± 3.44%, P < 0.001, respectively). The results from immunohistochemistry, Western blot, and real-time RT-PCR showed that the protein expression of Cx43 in the control group was significantly lower than that in the metoprolol and NC groups in the infarcted zone. CONCLUSIONS: NC decreased the incidence of spontaneous ventricular arrhythmias (especially VF), reduced Cx43 degradation, and improved Cx43 redistribution in myocardial infarcted zone in rats with hyper-AMI. The data of the present study indicated that NC may be a promising drug in the future to prevent patients with AMI from lethal ventricular arrhythmias in prehospital setting.

An approach based on antioxidant fingerprint-efficacy relationship and TLC bioautography assay to quality evaluation of Rubia cordifolia from various sources.

In this work, an approach based on antioxidant fingerprint-efficacy relationships and TLC bioautography assay was developed for quality evaluation of traditional Chinese medicine (TCM). First, chemical fingerprints of 20 batches of different sources of Rubia cordifolia were established by HPLC. The antioxidant activities of these samples were evaluated by 1,1-diphenyl-2-picrylhydrazyl radical scavenging assay. Then, five active components from fingerprint peaks were determined by both multiple correlation analysis and TLC bioautography assay. Similarity analysis and hierarchical clustering analysis (HCA) based on these active peaks were chosen to evaluate the quality of R. cordifolia from different sources. The samples with similarities below 0.9 had poorer antioxidant activities and those having stronger activities fell into the same cluster in HCA. Finally, five batches of commercial samples were processed in the same way to verify the feasibility of this method. The results suggested that chemical fingerprinting combined with similarity analysis based on fingerprint-efficacy relationship and TLC bioautography could evaluate the antioxidant activity of R. cordifolia from various sources. The established technique would also provide an easy method of evaluating the quality of other TCMs.

[Correlation study of auto-immune antibodies and rheumatoid arthritis patients of Shen deficiency syndrome].

OBJECTIVE: To investigate the correlation between auto-immune antibodies and rheumatoid arthritis (RA) patients of Shen deficiency syndrome (SDS), thus providing clinical evidence for further researches on molecular biological mechanisms of RA patients of SDS. METHODS: Totally 451 RA patients were assigned to the SDS group and the non-SDS group. Their general conditions (including gender, age, duration, and age of onset), C reactive protein (CRP), erythrocyte sedimentation rate (ESR), platelet (PLT), disease activities (DAS28), auto-antibodies [rheumatoid factor (RF), anti-CCP antibodies, anti-nuclear antibody (ANA)] were compared between the two groups. RESULTS: (1) The scores for EST, PLT, and DAS28 were obviously higher in the SDS group than in the non-SDS group (P < 0.05, P <0. 01). (2) The level of average RF was (697.32 +/-1 061.38 IU/mL) in the SDS group, higher than that in the non-SDS group (439.91 +/- 672.24 IU/mL, P <0.01). There was no statistical difference in anti-CCP antibody between the two groups (P >0.05).(3) The ANA positive rate of RA patients was 29. 63% (120/405). It was 37.19% (74/199) in RA patients of SDS and 22. 33% (46/206) in RA patients of non-SDS, showing statistical difference between the two groups (P <0.01). (4) The odds ratio for high level RF positive and ANA positive was 1. 574 and 2. 059 folds in RA patients of SDS as high as that in RA patients of non-SDS. CONCLUSIONS: RA patients of SDS would have higher risk of having auto-immune antibodies, fastened development, more worsen joint damage, and more poor prognosis. Its mechanisms might be closely associated with autoimmune tolerance.

[Pharmacological mechanisms of ethyl acetate extraction of Tongbiheji for active T cell signaling molecules].

AIM: To confirm extracts of activity from Traditional Chinese Medicine TongBiHeJi, study effect on two signaling pathways of T cells and clarify the pharmacological mechanisms of TongBiHeJi. METHODS: Concanavalin(ConA) were added successively into rats lymphocytic culture with different extracts of activity from Traditional Chinese Medicine. After 24 hours, CD71 expression rate on rat T lymphocytes activated with ConA was analyzed by flow cytometry. TCR, CD28 and ICOS on T cells were detected after T lymphocytes of rat activated by ConA were cultivated with various EthylAcetate extraction of TongBiHeJi(TBHJ) and Methotrexate (MTX) for 48 hours. RESULTS: CD71 expression rate on rat T lymphocytes induced by ConA was increased to 69.7%. TBHJ inhibited the rate of CD3(+);CD71(+); expression(32.5%); ConA up-regulated TCR, CD28 and ICOS expression on T cells obviously. There was different between ConA and positive control significantly(P<0.001). TBHJ could down-regulate obviously TCR, CD28 and ICOS expression on ConA-activated T lymphocytes with Concentration-dependent, especially ICOS. MTX inhibited CD3(+);CD71(+); and CD3(+);TCR(+); expression also. CONCLUSION: TBHJ inhibited T cells activation by adjusting two signaling pathways. That implied TBHJ could block CD28-ICOS signaling molecules to induce immunological tolerance. This study provided an experimental basis for application of TongBiHeJi to treatment of rheumatoid arthritis.

Liver targeting effect of vinegar-baked Radix Bupleuri on rhein in rats.

Vinegar-baked Radix Bupleuri (VBRB) is usually used to focus other drugs effect on liver in traditional Chinese medicine (TCM). However, no sufficient scientific data are available to support this concept. In this paper, the liver targeting enhancing effect of VBRB on rhein was investigated. 432 of rats were divided into two large groups according to the dose of rhein, low dose group of rhein (LDGR) and high dose group of rhein (HDGR). In each group, the rats were further divided into four subgroups, rhein control and rhein co-administered with three different doses of VBRB peroral. Concentrations of rhein and its metabolite in different tissues were determined by HPLC. Compared to the control group, VBRB significantly increased the distribution of both rhein and its metabolite in liver and meanwhile decreased their distribution in other tissues, indicating a strong liver targeting enhancing effect. This liver targeting effect of VBRB depended on the dose of VBRB and rhein. Low and high dose of VBRB had a more strong effect than medium dose in HDGR; high dose of rhein was more sensitive than low dose of rhein (P<0.05). Rhein existed in two forms after peroral administration in vivo. It was found that the liver targeting effect of VBRB was more remarkable with the native form of rhein compared to its derivative form. The results of this paper demonstrated that co-administration with VBRB is a simple and efficiencient method for liver targeting therapy, and the meridine guide theory of TCM was credible.

Effects of Ginkgo biloba extract on the pharmacokinetics of bupropion in healthy volunteers.

AIMS: To assess the effects of Ginkgo biloba extract on the pharmacokinetics of bupropion in healthy volunteers. METHODS: Fourteen healthy male volunteers (age range 19-25 years) received orally administered bupropion (150 mg) alone and during treatment with G. biloba 240 mg day(-1) (two 60-mg capsules taken twice daily) for 14 days. Serial blood samples were obtained over 72 h after each bupropion dose, and used to derive pharmacokinetic parameters of bupropion and its CYP2B6-catalysed metabolite, hydroxybupropion. RESULTS: Ginkgo biloba extract administration resulted in no significant effects on the AUC(0-infinity) of bupropion and hydroxybupropion. Bupropion mean AUC(0-infinity) value was 1.4 microg.h ml(-1)[95% confidence interval (CI) 1.2, 1.6] prior to G. biloba treatment and 1.2 microg.h ml(-1) (95% CI 1.1, 1.4) after 14 days of treatment. Hydroxybupropion mean AUC(0-infinity) value was 8.2 microg.h ml(-1) (95% CI 6.5, 10.4) before G. biloba administration and 8.7 microg.h ml(-1) (95% CI 7.1, 10.6) after treatment. The C(max) of hydroxybupropion increased from 221.8 ng ml(-1) (95% CI 176.6, 278.6) to 272.7 ng ml(-1) (95% CI 215.0, 345.8) (P = 0.038) and the t(1/2) of hydroxybupropion fell from 25.0 h (95% CI 22.7, 27.5) to 21.9 h (95% CI 19.9, 24.1) (P = 0.000). CONCLUSIONS: Ginkgo biloba extract administration for 14 days does not significantly alter the basic pharmacokinetic parameters of bupropion in healthy volunteers. Although G. biloba extract treatment appears to reduce significantly the t(1/2) and increase the C(max) of hydroxybupropion, no bupropion dose adjustments appear warranted when the drug is administered orally with G. biloba extract, due to the lack of significant change observed in AUC for either bupropion or hydroxybupropion.

[Intervenient effect of compound traditional Chinese medicine on dendritic cells in peripheral blood in vitro].

AIM: To investigate the intervenient effect of compound traditional Chinese medicine on dendritic cells (DCs) in peripheral blood in vitro. METHODS: DCs in peripheral blood were cultured for five days with medium which had GM-CSF and IL-4. The compound traditional Chinese medicine were added into the medium and the cells were collected on the 5th day. Surface markers of CD83 and CD86 in DCs were analyzed by flow cytometry. The capability of DC to stimulate the proliferation of T lymphocytes was evaluated by allogeneic mixed lymphocyte reaction. The content of IL-12 in cell culture supernatant was detected by ELISA. RESULTS: The expression of CD83 and CD86 was increased significantly(P<0.001) after the addition of the compound traditional Chinese medicine. The capability of DC to stimulate the proliferation of T lymphocytes was increased evidently (P<0.05). However the production of IL-12 was decreased obviously(P<0.001). CONCLUSION: The compound traditional Chinese medicine has immunoregulation of DC, It can enhance the presenting capability of antigen in DCs and inhibit the production of IL-12.

[Studies on the changes of 5-hydroxymethyl furfuraldehyde content in Radix Rehmanniae steamed for different time].

OBJECTIVE: To find the changing regularity of 5-Hydroxymethyl furfuraldehyde (5-HMF) content in Radix Rehmanniae steamed for different time. METHOD: HPLC method was used, with C18 column (6 mm x 200 mm, 5 microm), a mixture of methanol-water (15: 85) as the mobile phase. The flow rate was 1.0 mL x min(-1). The UV detection was setat 284 nm and the column temperature was set at 24 degrees C. RESULT: The linear range was from 0.01 to 0.08 microg, r = 0.999 9 (n = 5). The average recovery was 100.2% and RSD was 2.7% (n = 5). CONCLUSION: The 5-HMF content increases as the processing time prolongs in a certain time rang.