Time - sensitive effects of Zhuang medicated thread moxibustion on estrogen level in female perimenopausal model rats / Efectos sensibles al tiempo de la moxibustión con hilo medicado de Zhuang sobre el nivel de estrógeno en ratas modelo perimenopáusicas hembras

The present study was conducted to ascertain the estrogenic effect of Zhuang Medicated Thread Moxibustion (ZMTM) and explore its time - sensitive impact on estradiol (E2) in female perimenopausal rats. 40 female rats were randomized into four gr oups: the control, model, ZMTM, and acupuncture groups. The perimenopausal syndrome was induced in the last three groups with a daily subcutaneous dose of 80 mg/kg of 4 - vinylcyclohexene diepoxide for 15 days. Afterward, rats in the model and control group s were fed routinely, while rats in the ZMTM and acupuncture groups were treated with six ZMTM and acupuncture courses, respectively. Results of the study suggested that following the six courses of treatment, the E2 level in the model group was significan tly the lowest, while the regular group was the highest (P < 0.05). There was also a gradual increase in the E2 level of the ZMTM group compared to the model and acupuncture groups, e.g. after the 5th and 6th courses of treatment, their E2 level was signif icantly higher than the model and acupuncture groups. The ZMTM group was better than the model and acupuncture groups. In summary, ZMTM can improve perimenopausal induced rats' estrogen level.

El presen te estudio se llevó a cabo para determinar el efecto estrogénico de la moxibustión con hilo medicado Zhuang (ZMTM) y explorar su impacto sensible al tiempo en el estradiol (E2) en ratas hembras perimenopáusicas. Se dividió al azar una muestra de 40 ratas h embras en cuatro grupos: control, modelo, ZMTM y acupuntura. El síndrome perimenopáusico se indujo en los últimos tres grupos con una dosis subcutánea diaria de 80 mg/kg de diepóxido de 4 - vinilciclohexeno durante 15 días. Después, las ratas en los grupos m odelo y control fueron alimentadas rutinariamente, mientras que las ratas en los grupos ZMTM y acupuntura recibieron seis cursos de ZMTM y acupuntura, respectivamente. Los resultados del estudio sugieren que después de los seis cursos de tratamiento, el ni vel de E2 en el grupo modelo fue significativamente más bajo, mientras que el grupo regular fue más alto ( p < 0,05). También hubo un aumento gradual en el nivel de E2 del grupo ZMTM en comparación con los grupos modelo y acupuntura, por ejemplo, desp ués del quinto y sexto cursos de tratamiento, su nivel de E2 fue significativamente más alto que los grupos modelo y acupuntura. El grupo ZMTM fue mejor que los grupos modelo y acupuntura. En resumen, el ZMTM puede mejorar el nivel de estrógeno de las rata s inducidas por la perimenopausia.

[Effects of acupuncture on neurologic function and serum inflammatory factors after thrombolysis in acute ischemic stroke].

OBJECTIVE: To observe the effects of acupuncture on neurologic function and serum inflammatory factors in patients after thrombolysis in acute ischemic stroke (AIS). METHODS: A total of 102 AIS patients with onset to treatment time (OTT) ≤3 h were randomly divided into an observation group and a control group, 51 cases each group. In the control group, thrombolysis and conventional medical treatment were applied. On the basis of the treatment as the control group, acupuncture at Shuigou (GV 26), Zhongwan (CV 12), Qihai (CV 6), Neiguan (PC 6), etc. was applied in the observation group, 30 min each time, once a day. Both groups were treated for 2 weeks. Before and after treatment, the scores of National Institutes of Health stroke scale (NIHSS), modified Rankin scale (mRS), modified Barthel index (MBI) and serum level of homocysteine (Hcy), hypersensitive C-reactive protein (hs-CRP) were compared, and the clinical efficacy was evaluated in the two groups. RESULTS: After treatment, the scores of NIHSS, mRS and serum level of Hcy, hs-CRP were decreased compared with those before treatment (P<0.05), while the MBI scores were increased (P<0.05) in the two groups. The scores of NIHSS, mRS and serum level of Hcy, hs-CRP in the observation group were lower than those in the control group (P<0.05, P<0.01), the MBI score in the observation group was higher than that in the control group (P<0.01). The total effective rate was 88.2% (45/51) in the observation group, which was superior to 70.6% (36/51) in the control group (P<0.05). CONCLUSION: Acupuncture could promote the recovery of neurologic function in patients after thrombolysis in AIS, improve the ability of daily living, which may be related to reducing the level of inflammatory factors, thus inhibiting inflammatory response and improving cerebral ischemia reperfusion injury.

Research progress on chemical constituents and pharmacological effects of Ajania plants / 中国中药杂志

Ajania belonging to the subtribe Artemisiinae of Anthemideae(Asteraceae) is a genus of semi-shrubs closely related to Chrysanthemum. There are 24 species of Ajania in northwestern China, most of which are folk herbal medicines with strong stress tolerance. Modern medical studies have demonstrated that the chemical constituents of Ajania mainly include terpenoids, flavonoids, phenylpropanoids, alkynes, and essential oils. These compounds endow the plants with antimicrobial, anti-inflammatory, antitumor, antimalarial, antioxidant, and insecticide effects. In this study, we reviewed the research progress in the chemical constituents and pharmacological activities of Ajania, aiming to provide reference for the further research and development of Ajania.

Heat stress affects potato's volatile emissions that mediate agronomically important trophic interactions.

Potato, a cool-weather crop, emits volatile organic compounds (VOCs) which attract the specialist herbivore, Phthorimaea operculella, but also this herbivore's parasitic wasp, Trichogramma chilonis, an important biocontrol agent. What happens to this trophic system when heat stress challenges this agro-ecosystem? We studied how high temperature (HT) pre-treatments influence potato's VOC emissions and their subsequent effects on the preferences of insects, as evaluated in oviposition assays and Y-tube olfactometers. HT pre-stressed plants were less attractive to P. operculella adult moths, which were repelled by HT VOCs, but increased the recruitment of the parasitoid, T. chilonis, which were attracted. VOC emissions, including the most abundant constituent, ß-caryophyllene, were enhanced by HT treatments; some constituents elicited stronger behavioural responses than others. Transcripts of many genes in the biosynthetic pathways of these VOCs were significantly enhanced by HT treatment, suggesting increases in de novo biosynthesis. HT increased the plant's stomatal apertures, and exogenous applications of the hormone, ABA, known to suppress stomatal apertures, reduced leaf volatile emissions and affected the HT-altered plant attractions to both insects. From these results, we infer that HT stress affects this plant-insect interaction through its influence on VOC emissions, potentially decreasing herbivore ovipositions while increasing ovipositions of the parasitoid.

Kinsenoside alleviates inflammation and fibrosis in experimental NASH mice by suppressing the NF-κB/NLRP3 signaling pathway.

BACKGROUND: Non-alcoholic steatohepatitis (NASH) has replaced viral hepatitis as the main driver of the rising morbidity and mortality associated with cirrhosis and liver cancer worldwide, while no FDA-approved therapies are currently known. Kinsenoside (KD), naturally isolated from Anoectochilus roxburghii, possesses multiple biological activities, including lipolysis, anti-inflammation, and hepatoprotection. However, the effects of KD on NASH remain unclear. PURPOSE: This study aimed to explore the roles of KD in NASH and its engaged mechanisms. METHODS: Two typical animal models of NASH, mice fed a methionine-choline-deficient (MCD) diet (representing non-obese NASH) and mice fed a high-fat and -fructose diet (HFFD) (representing obese NASH), were used to investigate the effect of KD on NASH in vivo. Transcriptome sequencing was performed to elucidate the underlying mechanisms of KD. Lipopolysaccharide (LPS)-stimulated THP-1 cells and transforming growth factor ß1 (TGF-ß1)-activated LX-2 cells were applied to further explore the effects and mechanisms of KD in vitro. RESULTS: The intragastric administration of KD remarkably alleviated MCD/HFFD-induced murine NASH almost in a dose-dependent manner. Specifically, KD reduced lipid accumulation, inflammation, and fibrosis in the liver of NASH mice. KD ameliorated alanine aminotransferase (ALT), aspartate aminotransferase (AST), superoxide dismutase (SOD), and malondialdehyde (MDA) abnormalities. In addition, it decreased the level of serum proinflammatory factors (IL-12p70, IL-6, TNF-α, MCP-1, IFN-γ) and the hepatic expression of typical fibrosis-related molecules (α-SMA, Col-I, TIMP-1). Mechanically, KD attenuated the MCD/HFFD-induced NASH through the inhibition of the NF-κB/NLRP3 signaling pathway. Consistently, KD reduced inflammation stimulated by LPS in THP-1 cells via suppressing the NF-κB/NLRP3 pathway. Furthermore, it prevented the activation of LX-2 cells directly, by inhibiting the proliferation stimulated by TGF-ß1, and indirectly, by inactivating the NLRP3 inflammasome in macrophages. CONCLUSION: For the first time, the practical improvement of NASH by KD was revealed. Our study found that KD exerted its alleviative effects on NASH through the inhibition of the NF-κB/NLRP3 signaling pathway. Given its hepatoprotective and nontoxic properties, KD has the potential to be a novel and effective drug to treat NASH.

[Phosphorus forms in marsh soils with different years of Spartina alterniflora invasion in the Minjiang River estuary, China]. / é—½æ±Ÿæ²³å£äº’èŠ±ç±³è‰ä¸åŒå ¥ä¾µå¹´é™æ¹¿åœ°åœŸå£¤ç£·èµ‹å­˜å½¢æ€.

We examined the effects of Spartina alterniflora invasion on phosphorus forms of marsh soils, based on the method of space-for-time substitution by selecting S. alterniflora marshes with different invasion years (SA1, 5-6 years; SA2, 8-10 years; and SA3, 12-14 years) in Shanyutan of the Minjiang River estuary. The results showed that in marsh soils of different invasion years, the proportion of hardly decomposable phosphorus (HCl-Pi and Residual-P) was the highest (46.4%-46.7%), followed by moderately decomposable phosphorus (NaOH-Pi, NaOH-Po and Sonic-Pi) (40.0%-44.0%), and the easily decomposable phosphorus (Resin-Pi, NaHCO3-Pi and NaHCO3-Po) was the lowest (9.5%-13.3%). With increasing invasion years of S. alterniflora, soil phosphorus forms and their spatial distributions were greatly altered. The contents of moderately decomposable phosphorus, hardly decomposable phosphorus, and total phosphorus (TP) generally increased, while easily decomposable phosphorus content generally decreased. Compared with SA1, the contents of moderately decomposable phosphorus, hardly decomposable phosphorus and TP in SA2 increased by 11.5%, 9.7% and 10.5%, while those in SA3 increased by 24.8%, 13.2% and 13.5%, respectively. The distribution of phosphorus forms was greatly altered with increasing invasion years, which was dependent on the variations of key factors such as EC, pH value and grain composition. The implementation of regular mowing activities for S. alterniflora in the Minjiang River estuary in recent years, to some extent, reduced the return of phosphorus from residues to soils and decreased the availability of the easily decomposable phosphorus in soils.

Community structure of bacterioplankton and its relationship with environmental factors in the upper reaches of the Heihe River in Qinghai Plateau.

Planktonic microorganisms play a key role in the biogeochemical processes of the aquatic system, and they may be affected by many factors. High-throughput sequencing technology was used in this study to investigate and study the bacterioplankton community of water bodies in the upper reaches of the Heihe River Basin in Qinghai Plateau. Results showed that Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria are the predominant phyla in this river section, while the main genera are Thiomonas, Acidibacillus, Acidocella, Rhodanobacter, Acidithiobacter and Gallionella, which are autochthonous in the acid-mine drainage. Additionally, total nitrogen, total phosphorus, permanganate index and pH are significantly correlated with the bacterioplankton abundance and are the main limiting factors for the spatial distribution of the bacterioplankton. PICRUSt inferred that the mainstream microbial assemblages had a higher abundance of KOs belong to metabolism of terpenoids and polyketides, while the tributary had higher abundance of KOs belong to the immune system. The relationship between bacterioplankton community composition and environmental factors in the Heihe River basin was discussed for the first time in this study, which provides a theoretical basis for the healthy, orderly development of the water environment in the Heihe River Basin.

Mechanism of synergistic DNA damage induced by caffeic acid phenethyl ester (CAPE) and Cu(II): Competitive binding between CAPE and DNA with Cu(II)/Cu(I).

Caffeic acid phenethyl ester (CAPE) is an active polyphenol of propolis from honeybee hives, and exhibits antioxidant and interesting pharmacological activities. However, in this study, we found that in the presence of Cu(II), CAPE exhibited pro-oxidative rather than antioxidant effect: synergistic DNA damage was induced by the combination of CAPE and Cu(II) together as measured by strand breakage in plasmid DNA and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) formation, which is dependent on the molar ratio of CAPE:Cu(II). Production of Cu(I) and H2O2 from the redox reaction between CAPE and Cu(II), and subsequent OH formation was found to be responsible for the synergistic DNA damage. DNA sequencing investigations provided more direct evidence that CAPE/Cu(II) caused preferential cleavage at guanine, thymine and cytosine residues. Interestingly, we found there are competitive binding between CAPE and DNA with Cu(II)/Cu(I), which changed the redox activity of Cu(II)/Cu(I), via complementary applications of different analytical methods. The observed DNA damage was mainly attributed to the formation of DNA-Cu(II)/Cu(I) complexes, which is still redox active and initiated the redox reaction near the binding site between copper and DNA. Based on these data, we proposed that the synergistic DNA damage induced by CAPE/Cu(II) might be due to the competitive binding between CAPE and DNA with Cu, and site-specific production of OH near the binding site of copper with DNA. Our findings may have broad biological implications for future research on the pro-oxidative effects of phenolic compounds in the presence of transition metals.

Keap1-Nrf2 signaling activation by Bardoxolone-methyl ameliorates high glucose-induced oxidative injury in human umbilical vein endothelial cells.

In cultured human umbilical vein endothelial cells (HUVECs) high glucose (HG) stimulation will lead to significant cell death. Bardoxolone-methyl (BARD) is a NF-E2 p45-related factor 2 (Nrf2) agonist. In this study we show that BARD, at only nM concentrations, activated Nrf2 signaling in HUVECs. BARD induced Keap1-Nrf2 disassociation, Nrf2 protein stabilization and nuclear translocation, increasing expression of antioxidant response element (ARE) genes. BARD pretreatment in HUVECs inhibited HG-induced reactive oxygen species production, oxidative injury and cell apoptosis. Nrf2 shRNA or knockout (using a CRISPR/Cas9 construct) reversed BARD-induced cytoprotection in HG-stimulated HUVECs. Conversely, forced activation of Nrf2 cascade by Keap1 shRNA mimicked BARD's activity and protected HUVECs from HG. Importantly, BARD failed to offer further cytoprotection against HG in the Keap1-silened HUVECs. Taken together, Keap1-Nrf2 cascade activation by BARD protects HUVECs from HG-induced oxidative injury.

[Effects of spatial expansion of Phragmites australis and Cyperus malaccensis on distributions and stocks of phosphorus in marshes in the Minjiang River estuary, China.] / 闽江河口芦苇与短叶茳芏空间扩展对湿地系统磷赋存特征的影响.

To investigate the distributions and stocks of total phosphorus (TP) in plant-soil systems of marshes in Shanyutan of the Minjiang River estuary and explore its influencing factors, Phragmites australis (PA) marsh and Cyperus malaccensis (CM) marsh before spatial expansion and ecotonal marsh (EM, P. australis and C. malaccensis in EM were denoted by PA' and CM', respectively) during spatial expansion were studied. Results showed that, as affected by spatial expansion, the contents of TP in both plant and soil in different marshes showed great variations. Compared with PA and CM marshes, soil TP in EM increased by 20.0% and 7.1%, respectively. The variation of soil TP in EM might be attributed to the alteration of soil particle composition, plant bio-mass and root/shoot ratio during spatial expansion. Except for leaves, TP in organs of P. australis in P. australis-C. malaccensis community (PA') was lower than that in C. malaccensis in P. australis-C. malaccensis community (CM'), due to the differences in absorption, utilization and translocation way of P among organs between the two species. The competition effect caused by spatial expansion greatly changed plant P allocation ratio of the two species. Compared with PA and CM, the allocations of P in roots and leaves of PA' increased, while only that in roots of CM' rose. During spatial expansion, the two species might adopt different adaptation strategies for P absorption and utilization to maintain their competitiveness. The PA might compete primarily by strengthening the P accumulation capacities of its roots and promoting leaf photosynthesis, whereas the CM might resist the spatial expansion of PA by increasing its underground biomass to enhance P absorption.

Molecular mechanism for the activation of the anti-tuberculosis drug isoniazid by Mn(III): First detection and unequivocal identification of the critical N-centered isoniazidyl radical and its exact location.

Isoniazid (INH), the most-widely used anti-tuberculosis drug, has been shown to be activated by Mn(III) to produce the reactive carbon-centered isonicotinic acyl radical, which was considered to be responsible for its anti-tuberculosis activity. However, it is still not clear whether the previously-proposed N-centered isoniazidyl radical intermediate can be initially produced or not; and if so, what is its exact location on the hydrazine group, distal- or proximal-nitrogen? Through complementary applications of ESR spin-trapping and HPLC/MS methods, here we show that the characteristic and transient N-centered isoniazidyl radical intermediate can be detected and identified from INH activation uniquely by Mn(III)Acetate not by Mn(III) pyrophosphate. The exact location of the radical was found to be at the distal-nitrogen of the hydrazine group by 15N-isotope-labeling techniques via using 15N-labeled INH. Diisonicotinyl hydrazine was identified as a new reaction product from INH/Mn(III). Analogous results were observed with other hydrazides. This study represents the first detection and unequivocal identification of the initial N-centered isoniazidyl radical and its exact location. These findings should provide a new perspective on the molecular mechanism of INH activation, which may have broad biomedical and toxicological significance for future research for more efficient hydrazide anti-tuberculosis drugs.

1,25-Dihydroxyvitamin D exerts an antiaging role by activation of Nrf2-antioxidant signaling and inactivation of p16/p53-senescence signaling.

We tested the hypothesis that 1,25-dihydroxyvitamin D3 [1α,25(OH)2 D3 ] has antiaging effects via upregulating nuclear factor (erythroid-derived 2)-like 2 (Nrf2), reducing reactive oxygen species (ROS), decreasing DNA damage, reducing p16/Rb and p53/p21 signaling, increasing cell proliferation, and reducing cellular senescence and the senescence-associated secretory phenotype (SASP). We demonstrated that 1,25(OH)2 D3 -deficient [1α(OH)ase-/- ] mice survived on average for only 3 months. Increased tissue oxidative stress and DNA damage, downregulated Bmi1 and upregulated p16, p53 and p21 expression levels, reduced cell proliferation, and induced cell senescence and the senescence-associated secretory phenotype (SASP) were observed. Supplementation of 1α(OH)ase-/- mice with dietary calcium and phosphate, which normalized serum calcium and phosphorus, prolonged their average lifespan to more than 8 months with reduced oxidative stress and cellular senescence and SASP. However, supplementation with exogenous 1,25(OH)2 D3 or with combined calcium/phosphate and the antioxidant N-acetyl-l-cysteine prolonged their average lifespan to more than 16 months and nearly 14 months, respectively, largely rescuing the aging phenotypes. We demonstrated that 1,25(OH)2 D3 exerted an antioxidant role by transcriptional regulation of Nrf2 via the vitamin D receptor (VDR). Homozygous ablation of p16 or heterozygous ablation of p53 prolonged the average lifespan of 1α(OH)ase-/- mice on the normal diet from 3 to 6 months by enhancing cell proliferative ability and reducing cell senescence or apoptosis. This study suggests that 1,25(OH)2 D3 plays a role in delaying aging by upregulating Nrf2, inhibiting oxidative stress and DNA damage，inactivating p53-p21 and p16-Rb signaling pathways, and inhibiting cell senescence and SASP.

The efficacy and safety of Jiedu Tongluo granules for treating post-stroke depression with qi deficiency and blood stasis syndrome: study protocol for a randomized controlled trial.

BACKGROUND: Post-stroke depression (PSD) is the most common psychiatric complication after a stroke. The most frequently used antidepressants are selective serotonin receptor inhibitors (SSRIs) and serotonin and norepinephrine reuptake inhibitors (SNRIs), however, these exhibit a series of side effects. Traditional Chinese medicine has been used to treat PSD with few side effects. The aim of this study is to evaluate the efficacy and safety of Jiedu Tongluo granules for treating PSD with qi deficiency and blood stasis syndrome. METHODS: The planned study is a double-blind, randomized, placebo-controlled pilot trial. Eighty participants will be randomly assigned to receive either treatment or placebo. The treatment group will receive Jiedu Tongluo granules (JDTLG) with conventional treatment, and the placebo group will receive placebo with conventional treatment for 8 weeks. The primary outcome is the effectiveness of JDTLG on depression after 8 weeks treatment, which is defined as a decrease of 50% or more in 17-item Hamilton Depression Scale (HAMD-17) score or clinical recovery (score < 7). Secondary outcomes are improvement in neurological function, degree of independence, activities of daily living, and TCM syndrome at each visit, which will be measured with National Institute of Health Stroke Scale (NIHSS), modified Rankin Scale (mRS), Barthel Index (BI) and TCM scale, respectively. Interleukin (IL)-6, IL-8, and small-molecule metabolites will be monitored to explore the mechanism of action of JDTLG on PSD. Safety measures include vital signs, results of electrocardiography, laboratory index (full blood count, kidney and liver function tests) and adverse events. DISCUSSION: The purpose of this trial is to evaluate the therapeutic effects and safety of JDTLG in individuals with PSD with concomitant qi deficiency and blood stasis syndrome. If successful, the outcome of this trial will provide a viable treatment option for PSD patients. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT03147053 . Registered on 27 April 2017.

An efficient antimicrobial depot for infectious site-targeted chemo-photothermal therapy.

BACKGROUND: Silver and photothermal therapy (PTT) have been widely used for eradicating the drug-resistant bacteria. However, the risks of excess of silver for humans and the low efficiency of PTT still limit their in vivo therapeutic application. Integration of two distinctive bactericides into one entity is a promising platform to improve the efficiency of antimicrobial agents. RESULTS: In this study, a chemo-photothermal therapeutic platform based on polydopamine (PDA)-coated gold nanorods (GNRs) was developed. The PDA coating acquired high Ag+ ions loading efficiency and Cy5-SE fluorescent agent labeled glycol chitosan (GCS) conjugation (Ag+-GCS-PDA@GNRs). This platform became positively charged in the low pH environment of the abscess, allowing their accumulation in local infection site as revealed by thermal/florescence imaging. The loaded Ag+ ions was released in a pH-sensitive manner, resulting in selective Ag+ ions delivery to the abscess environment (pH ~ 6.3). More importantly, the ultralow dose of Ag+ ions could effectively damage the bacterial membrane, causing the permeability increase and the heat resistance reduction of the cell membrane, leading to the large improvement on bactericidal efficiency of PTT. On the other hand, the hyperthermia could trigger more Ag+ ions release, resulting in further improvement on bactericidal efficiency of chemotherapy. Combinational chemo-hyperthermia therapy of Ag+-GCS-PDA@GNRs could thoroughly ablate abscess and accelerate wound healing via a synergistic antibacterial effect. CONCLUSIONS: Our studies demonstrate that Ag+-GCS-PDA@GNRs is a robust and practical platform for use in chemo-thermal focal infection therapy with outstanding synergistic bacteria ablating.

Insulin resistance in vitamin D-deficient mice is alleviated by n-acetylcysteine.

Vitamin D deficiency will lead to insulin resistance. In the current study, vitamin D3 1α-Hydroxylase ["1α(OH)ase"] knockout mice were generated to mimic vitamin D deficiency in vivo. As compared to the wild-type mice, the liver tissues of the knockout mice showed impaired insulin signaling, decreased glucose transporter 4 expression and increased reactive oxygen species production. Meanwhile, p53-p21 activation, apoptosis intensity and pro-inflammatory cytokines (IL-6, IL-1 and MIP-1α) level were significantly increased in the knockout mice livers. Significantly, such effects in the knockout mice were largely attenuated by supplement with anti-oxidant n-acetylcysteine (NAC). Remarkably, insulin resistance and metabolic abnormalities in the knockout mice were largely alleviated after treatment of NAC. Therefore, inhibition of oxidative stress by NAC alleviated insulin resistance in vitamin D-deficient mice. Oxidative stress could be the primary cause of insulin resistance by vitamin D deficiency.

Mechanism of synergistic DNA damage induced by the hydroquinone metabolite of brominated phenolic environmental pollutants and Cu(II): Formation of DNA-Cu complex and site-specific production of hydroxyl radicals.

2,6-Dibromohydroquinone (2,6-DBrHQ) has been identified as an reactive metabolite of many brominated phenolic environmental pollutants such as tetrabromobisphenol-A (TBBPA), bromoxynil and 2,4,6-tribromophenol, and was also found as one of disinfection byproducts in drinking water. In this study, we found that the combination of 2,6-DBrHQ and Cu(II) together could induce synergistic DNA damage as measured by double strand breakage in plasmid DNA and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) formation, while either of them alone has no effect. 2,6-DBrHQ/Cu(II)-induced DNA damage could be inhibited by the Cu(I)-specific chelating agent bathocuproine disulfonate and catalase, but not by superoxide dismutase, nor by the typical hydroxyl radical (•OH) scavengers such as DMSO and mannitol. Interestingly, we found that Cu(II)/Cu(I) could be combined with DNA to form DNA-Cu(II)/Cu(I) complex by complementary application of low temperature direct ESR, circular dichroism, cyclic voltammetry and oxygen consumption methods; and the highly reactive •OH were produced synergistically by DNA-bound-Cu(I) with H2O2 produced by the redox reactions between 2,6-DBrHQ and Cu(II), which then immediately attack DNA in a site-specific manner as demonstrated by both fluorescent method and by ESR spin-trapping studies. Further DNA sequencing investigations provided more direct evidence that 2,6-DBrHQ/Cu(II) caused preferential cleavage at guanine, thymine and cytosine residues. Based on these data, we proposed that the synergistic DNA damage induced by 2,6-DBrHQ/Cu(II) might be due to the synergistic and site-specific production of •OH near the binding site of copper and DNA. Our findings may have broad biological and environmental implications for future research on the carcinogenic polyhalogenated phenolic compounds.

Effects of electroacupuncture intervention on expression of cyclooxygenase 2 and microglia in spinal cord in rat model of neuropathic pain / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of electroacupuncture (EA) treatment on the expression of cyclooxygenase (COX) 2 and microglia in spinal cord by using rat model of neuropathic pain, and to probe into the relationship between COX 2 and microglia.</p><p><b>METHODS</b>The rats were randomly divided into 6 groups, including normal control group, model group, sham group, EA 1 group (distant acupoints + local acupoints), EA 2 group (local acupoints), and EA 3 group (distant acupoints). Thermal withdrawal latencies were evaluated at 1 day preoperatively and 3, 5 and 7 days postoperatively. At 7 days postoperatively, the spinal COX 2 mRNA was detected by reverse-transcription polymerase chain reaction. Double immunofluorescent staining technology was applied to screen and verify the relationship between altered COX 2 and microglia.</p><p><b>RESULTS</b>Compared with the model group, thermal withdrawal latencies increased after EA treatment (P<0.01). The expressions of COX 2 mRNA were up-regulated in spinal cord of rat on day 7 after surgery (P<0.05). Compared with the model group, EA stimulation (EA 1 and EA 2 groups) reversed the up-regulation of COX 2 mRNA expression (P<0.05). EA 1 and EA 2 groups might have better treatment effect compared with the EA 3 group. Fluorescent images displayed COX 2 and microglia expressed at common areas.</p><p><b>CONCLUSIONS</b>EA was effective in analgesic and anti-inflammatory. EA has decreased the expression of spinal COX 2 mRNA in the trend of the therapeutic effect of "distant acupoints + local acupoints", and "local acupoints" intervention may be superior to that of "distant acupoints" intervention. Microglia may be related to the formation of COX 2.</p>

Therapeutic effect of acupuncture combining standard swallowing training for post-stroke dysphagia: A prospective cohort study.

OBJECTIVE: To assess the therapeutic effect of acupuncture combining standard swallowing training for patients with dysphagia after stroke. METHODS: A total of 105 consecutively admitted patients with post-stroke dysphagia in the Affiliated Hospital of Gansu University of Chinese Medicine were included: 50 patients from the Department of Neurology and Rehabilitation received standard swallowing training and acupuncture treatment (acupuncture group); 55 patients from the Department of Neurology received standard swallowing training only (control group). Participants in both groups received 5-day therapy per week for a 4-week period. The primary outcome measures included the scores of Videofluoroscopic Swallow Study (VFSS) and the Standardized Swallowing Assessment (SSA); the secondary outcome measure was the Royal Brisbane Hospital Outcome Measure for Swallowing (RBHOMS), all of which were assessed before and after the 4-week treatment. RESULTS: A total of 98 subjects completed the study (45 in the acupuncture group and 53 in the control group). Significant differences were seen in VFSS, SSA and RBHOMS scores in each group after 4-week treatment as compared with before treatment (P<0.01). Comparison between the groups after 4-week treatment showed that the VFSS P=0.007) and SSA scores (P=0.000) were more significantly improved in the acupuncture group than the control group. However, there was no statistical difference (P=0.710) between the acupuncture and the control groups in RBHOMS scores. CONCLUSIONS: Acupuncture combined with the standard swallowing training was an effective therapy for post-stroke dysphagia, and acupuncture therapy is worth further investigation in the treatment of post-stroke dysphagia.

Determination of endogenous brassinosteroids using sequential magnetic solid phase extraction followed by in situ derivatization/desorption method coupled with liquid chromatography-tandem mass spectrometry.

In this study, a sequential magnetic solid phase extraction followed by in situ derivatization/desorption method was proposed for the fast, selective and sensitive determination of brassinosteroids (BRs) in plant tissues. Magnetic sorbent for quick, easy, cheap, effective, rugged and safe method (mQuEChERS) and polymer(4-vinylphenylboronic acid-co-ethylene glycol dimethacrylate) coated Fe3O4@SiO2 (p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2) were prepared and characterized. Using them as sorbents, pigments and hydrophilic interferents were firstly removed from plant extract by mQuEChERS, and then endogenous BRs were selectively enriched by p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2 through boronate affinity interaction. After loading BRs on p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2, instead of directly eluting free BRs, the adsorbed BRs were released by adding 4-(N,N-dimethyamino)phenylboronic acid (4-DMAPBA) solution for in situ derivatizaiton/desorption of BRs based on a transesterification reaction between the boronate moieties of p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2 and 4-DMAPBA, finally the resultant solution was submitted to LC-MS/MS for quantification. The whole procedure of the sequential MSPE could be accomplished within 1h, and the matrix effect to MS signal after the sample pretreatment was estimated to be in the range of 93.0-97.4%. The established method provided broad linear dynamics ranges (1.0-100.0pg/mL) with correlation coefficients (R) >0.9978, substantial sensitivity (limits of detection ranged from 0.27 to 1.29pg/mL), high reproducibility (intra-day and inter-day relative standard deviations (RSDs) less than 14.8%) and satisfactory accuracy (recoveries ranged from 74.0%-116.6%). Furthermore, endogenous BRs were successfully detected in one flower of Brassica napus L. (22.5-542.7pg/g fresh weight) and other plant tissues (13.7-289.8pg/g fresh weight).

Antinociceptive effects of vitexin in a mouse model of postoperative pain.

Vitexin, a C-glycosylated flavone present in several medicinal herbs, has showed various pharmacological activities including antinociception. The present study investigated the antinociceptive effects of vitexin in a mouse model of postoperative pain. This model was prepared by making a surgical incision on the right hindpaw and von Frey filament test was used to assess mechanical hyperalgesia. Isobolographical analysis method was used to examine the interaction between vitexin and acetaminophen. A reliable mechanical hyperalgesia was observed at 2 h post-surgery and lasted for 4 days. Acute vitexin administration (3-10 mg/kg, i.p.) dose-dependently relieved this hyperalgesia, which was also observed from 1 to 3 days post-surgery during repeated daily treatment. However, repeated vitexin administration prior to surgery had no preventive value. The 10 mg/kg vitexin-induced antinociception was blocked by the opioid receptor antagonist naltrexone or the GABAA receptor antagonist bicuculline. The doses of vitexin used did not significantly suppress the locomotor activity. In addition, the combination of vitexin and acetaminophen produced an infra-additive effect in postoperative pain. Together, though vitexin-acetaminophen combination may not be useful for treating postoperative pain, vitexin exerts behaviorally-specific antinociception against postoperative pain mediated through opioid receptors and GABAA receptors, suggesting that vitexin may be useful for the control of postoperative pain.