Antiplatelet and myocardial protective effect of Shexiang Tongxin Dropping Pill in patients undergoing percutaneous coronary intervention: A randomized controlled trial.

BACKGROUND: High on-clopidogrel platelet reactivity could be partially explained by loss-of-function alleles of CYP2C19, the enzyme that converts clopidogrel into its active form. Shexiang Tongxin Dropping Pill (STDP) is a traditional Chinese medicine to treat angina pectoris. STDP has been shown to improve blood flow in patients with slow coronary flow and attenuate atherosclerosis in apolipoprotein E-deficient mice. However, whether STDP can affect platelet function remains unknown. OBJECTIVE: The purpose of this study is to examine the potential effects of STDP on platelet function in patients undergoing percutaneous coronary intervention (PCI) for unstable angina. The interaction between the effects of STDP with polymorphisms of CYP2C19 was also investigated. DESIGN, PARTICIPANTS AND INTERVENTION: This was a single-center, randomized controlled trial in patients undergoing elective PCI for unstable angina. Eligible subjects were randomized to receive STDP (210 mg per day) plus dual antiplatelet therapy (DAPT) with clopidogrel and aspirin or DAPT alone. MAIN OUTCOME MEASURES: The primary outcome was platelet function, reflected by adenosine diphosphate (ADP)-induced platelet aggregation and platelet microparticles (PMPs). The secondary outcomes were major adverse cardiovascular events (MACEs) including recurrent ischemia or myocardial infarction, repeat PCI and cardiac death; blood biomarkers for myocardial injury including creatine kinase-MB isoenzyme (CK-MB) and high-sensitive troponin I (hsTnI); and biomarkers for inflammation including intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1) and galectin-3. RESULTS: A total of 118 subjects (mean age: [66.8 ± 8.9] years; male: 59.8%) were included into analysis: 58 in the control group and 60 in the STDP group. CYP2C19 genotype distribution was comparable between the 2 groups. In comparison to the control group, the STDP group had significantly lower CK-MB (P < 0.05) but similar hsTnI (P > 0.05) at 24 h after PCI, lower ICAM-1, VCAM-1, MCP-1 and galectin-3 at 3 months (all P < 0.05) but not at 7 days after PCI (P > 0.05). At 3 months, the STDP group had lower PMP number ([42.9 ± 37.3] vs. [67.8 ± 53.1] counts/µL in the control group, P = 0.05). Subgroup analysis showed that STDP increased percentage inhibition of ADP-induced platelet aggregation only in slow metabolizers (66.0% ± 20.8% in STDP group vs. 36.0% ± 28.1% in the control group, P < 0.05), but not in intermediate or fast metabolizers. The rate of MACEs during the 3-month follow-up did not differ between the two groups. CONCLUSION: STDP produced antiplatelet, anti-inflammatory and cardioprotective effects. Subgroup analysis indicated that STDP inhibited residual platelet reactivity in slow metabolizers only. TRIAL REGISTRATION: This study was registered on www.chictr.org.cn: ChiCTR-IPR-16009785.

Kellerin from Ferula sinkiangensis exerts neuroprotective effects after focal cerebral ischemia in rats by inhibiting microglia-mediated inflammatory responses.

ETHNOPHARMACOLOGICAL RELEVANCE: Ferula sinkiangensis K. M. Shen is a traditional Chinese medicine that has a variety of pharmacological properties relevant to neurological disorders and inflammations. Kellerin, a novel compound extracted from Ferula sinkiangensis, exerts a strong anti-neuroinflammatory effect by inhibiting microglial activation. Microglial activation plays a vital role in ischemia-induced brain injury. However, the potential therapeutic effect of kellerin on focal cerebral ischemia is still unknown. AIM OF THE STUDY: To explore the effect of kellerin on cerebral ischemia and clarify its possible mechanisms, we applied the middle cerebral artery occlusion (MCAO) model and the LPS-activated microglia model in our study. MATERIALS AND METHODS: Neurological outcome was examined according to a 4-tiered grading system. Brain infarct size was measured using TTC staining. Brain edema was calculated using the wet weight minus dry weight method. Neuron damage and microglial activation were observed by immunofluorescence in MCAO model in rats. In in vitro studies, microglial activation was examined by flow cytometry and the viability of neuronal cells cultured in microglia-conditioned medium was measured using MTT assay. The levels of pro-inflammatory cytokines were measured by qRT-PCR and ELISA. The proteins involved in NF-κB signaling pathway were determined by western blot. Intracellular ROS was examined using DCFH-DA method and NADPH oxidase activity was measured using the NBT assay. RESULTS: We found that kellerin improved neurological outcome, reduced brain infarct size and decreased brain edema in MCAO model in rats. Under the pathologic conditions of focal cerebral ischemia, kellerin alleviated neuron damage and inhibited microglial activation. Moreover, in in vitro studies of LPS-stimulated BV2 cells kellerin protected neuronal cells from being damaged by inhibiting microglial activation. Kellerin also reduced the levels of pro-inflammatory cytokines, suppressed the NF-κB signaling pathway, and decreased ROS generation and NADPH oxidase activity. CONCLUSIONS: Our discoveries reveal that the neuroprotective effects of kellerin may largely depend on its inhibitory effect on microglial activation. This suggests that kellerin could serve as a novel anti-inflammatory agent which may have therapeutic effects in ischemic stroke.

Kellerin alleviates cognitive impairment in mice after ischemic stroke by multiple mechanisms.

Ischemic stroke is a global disease with high disability and mortality rates. Cognitive impairment is one of the major clinical features of ischemic stroke, and microglia-mediated inflammation has been shown to be an important contributor to the pathogenesis of ischemic stroke. Kellerin, extracted from Ferula sinkiangensis, was previously shown to inhibit microglial activation and exert a strong anti-neuroinflammatory effect. However, there is no report of the potential therapeutic effect of kellerin on ischemic stroke by targeting microglial cells. In this study, we wanted to examine the effects of kellerin on ischemic stroke in the bilateral common carotid artery occlusion (BCCAO) model and the lipopolysaccharide (LPS)-activated microglia model. We found that kellerin alleviated cognitive impairment, decreased neuronal loss, suppressed microglial activation, and transformed microglia from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype in BCCAO mice. Moreover, in in vitro studies, we found that kellerin regulated microglial polarization and inhibited the NLRP3 and MAPK signaling pathways after LPS treatment. These findings provide a new understanding of the function of kellerin in ischemic stroke, and suggest that kellerin could be a potential therapeutic agent for the treatment of ischemic stroke.

[Acupuncture combined with medication improves endocrine hormone levels and ovarian reserve function in poor ovarian response patients undergoing in vitro fertilization-embryo transplantation].

OBJECTIVE: To observe the clinical effect of acupuncture plus medication in the treatment of poor ovarian response (POR) patients and to explore its mechanisms in assisting pregnancy. METHODS: A total of 100 volunteer POR women undergoing in vitro fertilization-embryo transplantation (IVF-ET) were recruited in the present study. On the 1st cycle of IVF-ET, these POR women received microstimulation of ovulation program (oral administration of Clomiphene, muscular injection of Menotrophin, Chorionic Gonadotrophin triggering, etc.). Before receiving the 2nd period of IVF-ET, these patients were equally and randomly divided into control, medication (Climen, composed of estradiol valerate and cyproterone acetate), acupuncture and acupuncture+medication (combined treatment) groups according to the random number table. Patients of the medication group were asked to orally take Climen (1 tablet/d for 21 days) beginning from the 3rd day of the menstruation, which was repeated for 3 menstrual cycles. Patients of the acupuncture group received manual acupuncture stimulation of Guanyuan (CV4), and bilateral Taixi (KI3), Sanyinjiao (SP6) of and Tai-chong (LR3) from day 8 to 15 of menstruation (follicular phase), once daily for 3 menstrual cycles. On the 2nd day of menstruation of the 1st and 2nd IVF-ET cycle, the ovarian reserve function was detected, including measurement of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) contents by using radioimmunoassay, and serum anti-mullerian hormone (AMH) level by using ELISA, and the antral follicles count (AFC) of the ovaries by using a color Doppler ultrasonic diagnosis apparatus. At the end of ovulation induction, the assisted pregnancy indexes and outcomes were detected, including administration of dosage and days of gonadotropin (Gn), the diameter of dominant oocyte, level of E2 on the trigger day, the numbers of ultrasound-guided-retrieved oocyte and the cultivated high-quality embryo (grade 1 and 2). RESULTS: After the treatment, the contents of serum FSH, LH and E2 in the medication, acupuncture and combined treatment groups were significantly reduced (P<0.05), and the content of serum AMH and ovary AFC in the medication, acupuncture and combined treatment groups were obviously increased (P<0.05) in comparison with their own pre-treatment (P<0.05). Compared with the control group, the levels of FSH, LH and E2 contents were notably decreased, and the AMH content and AFC significantly increased in the medication, acupuncture and combined treatment groups (P<0.05). Comparison among the three treatment groups showed that the levels of FSH, LH and E2 were notably lower in the combined treatment group than in the acupuncture group (P<0.05), and the AFC was markedly more in the combined treatment group than in the acupuncture group (P<0.05), and LH content was also significantly lower in the combined treatment group than in the medication group (P<0.05). Regarding the pregnancy-assistant indexes and outcomes, the dosage of Gn in the combined treatment group and the number of Gn administration days in the acupuncture and combined treatment groups were significantly fewer (P<0.05), serum E2 levels on the trigger day in the medication, acupuncture and combined treatment groups, and the high-quality embryo number in the combined treatment group were considerably higher in comparison with their own levels of the 1st IVF-ET cycle in each group (P<0.05). Comparison among the three groups showed that the dosage and number of administration days of Gn in the combined treatment group were fewer than those in the medication group(P<0.05), and the dosage of Gn in the combined treatment group were significantly lower than that in the acupuncture group (P<0.05), E2 content of the combined treatment group was markedly higher than that of the medication and acupuncture groups (P<0.05). The high-quality embryo number in the combined treatment group was obviously larger than that of the medication group (P<0.05). No significant differences were found between the 2nd and 1st IVF-ET cycles in the number of the obtained oocyte, between the acupuncture and medication groups in the E2 content at the trigger day, and the numbers of Gn administration days and the high-quality embryo (P>0.05). CONCLUSION: Acupuncture combined with medication improves the level of endocrinal hormones and ovarian reservation function in POR women undergoing IVF-ET, benefiting the ovary environment of pregnancy.

Okanin, effective constituent of the flower tea Coreopsis tinctoria, attenuates LPS-induced microglial activation through inhibition of the TLR4/NF-κB signaling pathways.

The EtOAc extract of Coreopsis tinctoria Nutt. significantly inhibited LPS-induced nitric oxide (NO) production, as judged by the Griess reaction, and attenuated the LPS-induced elevation in iNOS, COX-2, IL-1ß, IL-6 and TNF-α mRNA levels, as determined by quantitative real-time PCR, when incubated with BV-2 microglial cells. Immunohistochemical results showed that the EtOAc extract significantly decreased the number of Iba-1-positive cells in the hippocampal region of LPS-treated mouse brains. The major effective constituent of the EtOAc extract, okanin, was further investigated. Okanin significantly suppressed LPS-induced iNOS expression and also inhibited IL-6 and TNF-α production and mRNA expression in LPS-stimulated BV-2 cells. Western blot analysis indicated that okanin suppressed LPS-induced activation of the NF-κB signaling pathway by inhibiting the phosphorylation of IκBα and decreasing the level of nuclear NF-κB p65 after LPS treatment. Immunofluorescence staining results showed that okanin inhibited the translocation of the NF-κB p65 subunit from the cytosol to the nucleus. Moreover, okanin significantly inhibited LPS-induced TLR4 expression in BV-2 cells. In summary, okanin attenuates LPS-induced activation of microglia. This effect may be associated with its capacity to inhibit the TLR4/NF-κB signaling pathways. These results suggest that okanin may have potential as a nutritional preventive strategy for neurodegenerative disorders.

Natural potential neuroinflammatory inhibitors from Alhagi sparsifolia Shap.

Neuroinflammation is a key contributor to neuronal damage in neurodegenerative diseases. In our previous work on natural effective neuroinflammatory inhibitors, Alhagi sparsifolia Shap. (Leguminosae), a folk medicine widely distributed in Xinjiang, attracted our attention because of its significant anti-neuroinflammatory effect. Therefore, further investigation of the bioactive material basis was carried out. As a result, 33 major components were characterized and identified by chromatographic and spectral methods, respectively. Furthermore, the anti-neuroinflammatory effects of the extract and purified constituents were evaluated in LPS-induced N9 cells in vitro. The results displayed that compounds 1, 2, 3, 5, 6, 8, 11, 15, 16, 17, 22, 23, 25, 26, 28, 30, 33 could exhibit significant inhibitory activities without obvious cytotoxicities at their effective concentrations. Especially, isorhamnetin (1) (IC50 17.87µM), quercetin (2) (10.22µM), 3',7-dihydroxyl-4'-methoxylisoflavone (5) (17.43µM), 3',7-dihydroxyl-4',6-dimethoxylisoflavone (6) (11.21µM), syringgaresinol (16) (2.68µM), bombasinol A (17) (7.61µM), aurantiamide (23) (14.91µM) and 1,3,3,4-tetramethyl cyclopentene (33) (2.63µM) showed much stronger inhibiting effect than that of the positive control minocycline (19.89µM). Therefore, the effective compositions might be responsible for the significant neuroinflammation inhibitory activities exhibited by the herb. Moreover, compounds 16 and 33 could be good leading compounds for the development of potential therapeutic agents against neurodegenerative diseases.

Sesquiterpene Coumarins from Ferula sinkiangensis Act as Neuroinflammation Inhibitors.

Neuroinflammation mediated by microglia cells plays a critical role in the development of Alzheimer's disease. To identify novel natural neuroinflammation inhibitors, a bioactivity-guided phytochemical research was performed on the traditional Chinese medicine "Awei", that exhibited a significant inhibitory effect on nitric oxide production in over-activated microglia cells. The research identified sixteen bioactive sesquiterpene coumarins (two new and fourteen known ones) in the effective extract of Ferula sinkiangensis. Further, the anti-neuroinflammatory activities in BV-2 microglial cells were evaluated by monitoring LPS-induced nitric oxide production. In conclusion, the major constituent, (3'S, 5'S, 8'R, 9'S, 10'R)-kellerin (1.5 %, w/w), should be responsible for the anti-neuroinflammatory effect exhibited by Awei. Furthermore, it might be a potential natural therapeutic agent for Alzheimer's disease. The research indicated moreover, that its primary mechanism is the inhibition of mRNA expression of the inflammatory cytokines nitric oxide, tumor necrosis factor-α, cyclooxygenase-2, interleukin-6 and interleukin-1ß.

Repeated-dose (28 days) oral toxicity study in rats of an antiacne formula (BC-AF) derived from plants.

To evaluate the safety of a formula (BC-AF) consisting of the extracts of danshen (Salvia miltiorrhiza), loquat leaf (Glycyrrhiza uralensis), and licorice (Eriobotrya japonica), a preliminary 28-day, repeated-dose oral toxicity study was performed in Sprague-Dawley rats. Eighty animals were divided into four groups, with each group comprising 10 male and 10 female rats. BC-AF was administered once-daily by oral gavage at doses of 0 (control), 2.5 (low), 5 (middle), and 10 (high) g/kg body weight successively for each group for 28 days, respectively. Rats in all groups were sacrificed on day 29, except half of the males and females in the high-dose group that were kept for an additional 2 weeks to observe any possible toxicity after drug withdrawal. In 4 weeks, there were no toxicity reactions or abnormal deaths in any animal groups. There was no significant difference, in comparison to the control group, in clinical signs, organ weights, hematological and serological parameters, or histopathologic findings. In conclusion, the 28-day repeated-dose oral toxicity study demonstrates that BC-AF produced no effects in either male or female rats following oral administration of up to 10 g/kg.

Effective pollinators of Asian sacred lotus (Nelumbo nucifera): contemporary pollinators may not reflect the historical pollination syndrome.

BACKGROUND AND AIMS: If stabilizing selection by pollinators is a prerequisite for pollinator-mediated floral evolution, spatiotemporal variation in the pollinator assemblage may confuse the plant-pollinator interaction in a given species. Here, effective pollinators in a living fossil plant Nelumbo nucifera (Nelumbonaceae) were examined to test whether beetles are major pollinators as predicted by its pollination syndrome. METHODS: Pollinators of N. nucifera were investigated in 11 wild populations and one cultivated population, and pollination experiments were conducted to examine the pollinating role of two major pollinators (bees and beetles) in three populations. KEY RESULTS: Lotus flowers are protogynous, bowl shaped and without nectar. The fragrant flowers can be self-heating during anthesis and produce around 1 million pollen grains per flower. It was found that bees and flies were the most frequent flower visitors in wild populations, contributing on average 87.9 and 49.4 % of seed set in Mishan and Lantian, respectively. Beetles were only found in one wild population and in the cultivated population, but the pollinator exclusion experiments showed that beetles were effective pollinators of Asian sacred lotus. CONCLUSIONS: This study indicated that in their pollinating role, beetles, probable pollinators for this thermoregulating plant, had been replaced by some generalist insects in the wild. This finding implies that contemporary pollinators may not reflect the pollination syndrome.

Flower thermoregulation facilitates fertilization in Asian sacred lotus.

BACKGROUND AND AIMS: The thermoregulatory flower of the Asian sacred lotus (Nelumbo nucifera) can maintain a relatively stable temperature despite great variations in ambient temperature during anthesis. The thermoregulation has been hypothesized to offer a direct energy reward for pollinators in lotus flowers. This study aims to examine whether the stable temperature maintained in the floral chamber influences the fertilization process and seed development. METHODS: An artificial refrigeration instrument was employed to cool flowers during the fertilization process and post-fertilization period in an experimental population. The effect of temperature on post-pollination events was also examined by removing petals in two field populations. KEY RESULTS: Treatments with low floral temperature did not reduce stigma receptivity or pollen viability in undehisced anthers. Low temperature during the fertilization period significantly decreased seed set per flower but low temperature during the phase of seed development had no effect, suggesting that temperature regulation by lotus flowers facilitated fertilization success. Hand-pollination treatments in two field populations indicated that seed set of flowers with petals removed was lower than that of intact flowers in north China, where ambient temperatures are low, but not in south China, confirming that reducing the temperature of carpels did influence post-pollination events. CONCLUSIONS: The experiments suggest that floral thermoregulation in lotus could enhance female reproductive success by facilitating fertilization.

[Effect of shenqi fuzheng injection on pre/post-operational change of argyrophilic-nucleolar organizer regions in peripheral T-lymphocyte in patients with gastric carcinoma].

OBJECTIVE: To investigate the pre-post operational change of argyrophilic-nucleolar organizer regions (Ag-NORs) in peripheral T-lymphocyte of patients with gastric carcinoma (GC), and to explore the effect of shenqi fuzheng injection(SFI) on it. METHODS: Eighty five patients were divided into two groups according to the operation performed was radical or non-radical, and the two groups were subdivided into two by additional intravenous dripping of SFI was given to them or not. The content of Ag-NORs in peripheral T-lymphocyte in all patients before and after operation as well as in 12 healthy subjects was determined. RESULTS: Content of Ag-NORs in GC patients was significantly lower than that in the healthy subject (P < 0.01), which significantly increased after patients underwent radical operation (P < 0. 01 or P < 0.05), especially in those treated with SFI (P < 0.01). While in patients underwent non-radical operation but not treated with SFI, it showed insignificant change after operation, however it did significantly increase in those treated with SFI (P < 0.05). CONCLUSION: The immune function of T lymphocyte was low in patients with gastric carcinoma, post-operational adjuvant treatment of SFI can significantly improve the cellular immunity of patients.

Reducing the effects of driving fatigue with magnitopuncture stimulation.

The purpose of this study was to assess the effects of reducing driving fatigue with magnitopuncture stimuli on Dazhui (DU14) point and Neiguan (PC6) points using heart rate (HR), reaction time (RT) testing, right rate (RR), critical flicker fusion frequency (CFF) and subjective evaluation. Forty healthy subjects were randomly divided into two groups: study and control groups. All subjects were required to be well rested before the experiment. The subjects were engaged in high speed driving at a constant vehicle velocity of 80 km/h continuously for 3h on a test course simulating an expressway. During the driving magnitopunctures (Haci Five Elements Needle, 250 mT, made by Haci Company limited) were applied to the Dazhui (DU14) point and Neiguan (PC6) points for the study group when the subject performed the task for 2.5h, and for the control group magnitopunctures were applied to non-acupuncture points during the same time session. The results of this study show a significant effect of magnitopuncture stimuli on RT, RR and CFF. Subjective evaluation also exhibited significant differences (P < 0.05) between the two groups after the driving task. The findings showed that magnitopuncture stimuli on DU14 point and PC6 points could reduce the effects of driving fatigue.

[The effect of magnitopuncture stimulation on HRV during simulated driving under vibration conditions].

The purpose of this study was to assess the effect of magnitopuncture stimuli for reducing driver mental stress and fatigue using power spectral analysis of the heart rate variability (HRV) and subjective evaluation. The experiments were divided into A-group and B-group. In both groups the subjects performed the simulator for 90 minutes under a vibration conditions with an erect sitting posture in a silent environment, and magnitopuncture was put on the acupoints when performing the task for one hour in A-group. In this study HRV exhibited a significant difference between the two groups after the simulating task (P < 0.05). A conclusion that magnitopuncture stimuli can reduce the driver mental stress and fatigue effectively was drawn.

Effect of magnitopuncture on sympathetic and parasympathetic nerve activities in healthy drivers--assessment by power spectrum analysis of heart rate variability.

The object of this study was to assess the effects of magnitopuncture applied to Dazhui (DU14) point and Neiguan (PC6) points on sympathetic and parasympathetic nerve activities by power spectrum analysis of heart rate variability in healthy drivers during simulated driving. Using power spectrum analysis, the low frequency (LF) and high frequency (HF) components of heart rate variability can be calculated reflecting the sympathetic and parasympathetic activity. The 40 healthy male subjects were randomly divided into two groups: A (study group) and B (control group). All subjects were required to be well rested before the experiment. The subjects of both groups were required to perform a simulated driving task for 3 h. During the driving, magnitopunctures were applied to the DU14 and PC6 points for A while the subject performed the task for 2.5 h, and for B magnitopunctures were applied to non-acupuncture points which were 1.5 cm away from the two acupuncture points respectively over the same time. Subjective response to a questionnaire was obtained after the simulated task in the two groups. At the end of the driving task the LF component in normalized units (NU) had decreased significantly ( P<0.05) indicating a reduced sympathetic nerve activity and the HF component (NU) increased significantly ( P<0.05) indicating a increased parasympathetic nerve activity for A compared with pre-stimulation while for B no significant differences were observed. There were significant group differences in LF (NU), HF (NU) and LF:HF at the end of the driving task ( P<0.05). It was concluded that a modulating effect of magnitopuncture on sympathetic and parasympathetic nerve activities in healthy subjects was associated with the acupuncture points. The findings represent physiological evidence that magnitopuncture may reduce mental fatigue in healthy drivers.