Improvement of Panax notoginseng saponin accumulation triggered by methyl jasmonate under arbuscular mycorrhizal fungi.

Panax notoginseng is a highly valued perennial medicinal herb plant in Yunnan Province, China, and the taproots are the main medicinal parts that are rich in active substances of P. notoginseng saponins. The main purpose of this study is to uncover the physiological and molecular mechanism of Panax notoginseng saponin accumulation triggered by methyl jasmonate (MeJA) under arbuscular mycorrhizal fungi (AMF) by determining physiological indices, high-throughput sequencing and correlation analysis. Physiological results showed that the biomass and saponin contents of P. notoginseng, the concentrations of jasmonic acids (JAs) and the key enzyme activities involved in notoginsenoside biosynthesis significantly increased under AMF or MeJA, but the interactive treatment of AMF and MeJA weakened the effect of AMF, suggesting that a high concentration of endogenous JA have inhibitory effect. Transcriptome sequencing results indicated that differential expressed genes (DEGs) involved in notoginsenoside and JA biosynthesis were significantly enriched in response to AMF induction, e.g., upregulated genes of diphosphocytidyl-2-C-methyl-d-erythritol kinases (ISPEs), cytochrome P450 monooxygenases (CYP450s)_and glycosyltransferases (GTs), while treatments AMF-MeJA and salicylhydroxamic acid (SHAM) decreased the abundance of these DEGs. Interestingly, a high correlation presented between any two of saponin contents, key enzyme activities and expression levels of DEGs. Taken together, the inoculation of AMF can improve the growth and saponin accumulation of P. notoginseng by strengthening the activities of key enzymes and the expression levels of encoding genes, in which the JA regulatory pathway is a key link. This study provides references for implementing ecological planting of P. notoginseng, improving saponin accumulation and illustrating the biosynthesis mechanism.

Degradation kinetics of vitamins in premixes for pig: effects of choline, high concentrations of copper and zinc, and storage time.

OBJECTIVE: The present work was undertaken to evaluate the effects of storage time, choline chloride, and high concentrations of Cu and Zn on the kinetic behavior of vitamin degradation during storage in two vitamin premixes and four vitamin-trace mineral (VTM) premixes. METHODS: Two vitamin premixes (with or without 160,000 mg/kg of choline) were stored at 25°C and 60% humidity. Besides, four VTM premixes were used to evaluate the effects of choline (0 vs 40,000 mg/kg) and trace minerals (low CuSO4+ZnO vs high CuSO4+ZnO) on vitamin stability in VTM premixes stored in room, and the VTM premixes were stored in room temperature at 22°C. Subsamples from each vitamin and VTM premix were collected at 0, 1, 2, 3, 6, and 12 months. The retention of vitamin A (VA), vitamin D3 (VD3), vitamin E (VE), vitamin K3 (VK3), vitamin B1 (VB1), vitamin B2 (VB2), vitamin B3 (VB3), vitamin B5 (VB5), and vitamin B6 (VB6) in vitamin premixes and VTM premixes during storage was determined. The stability of vitamins in vitamin premixes and VTM premixes was determined and reported as the residual vitamin activity (% of initial) at each sampling point. RESULTS: The effect of choline on VK3 retention was significant in vitamin premixes (p<0.05). The negative effect of storage time was significant for the retentions of VD3, VK3, VB1, VB2, VB5, and VB6 in vitamin premix (p<0.05). For VTM premixes, negative effect of storage time was significant (p<0.05) for the losses of vitamin in VTM premixes. Choline and high concentrations of Cu and Zn significantly increased VA, VK3, VB1, and VB2 loss during storage (p<0.05). The supplementation of high concentrations of Cu and Zn significantly decreased the concentrations of VD3 and VB6 (p<0.05) in VTM premixes at extended storage time. CONCLUSION: The maximum vitamin stability was detected in vitamin and VTM premixes containing no choline or excess Cu and Zn. The results indicated that extended storage time increased degradation of vitamin in vitamin or VTM premixes. These results may provide useful information for vitamin and VTM premixes to improve the knowledge of vitamin in terms of its stability.

The strategies for the supplementation of vitamins and trace minerals in pig production: surveying major producers in China.

OBJECTIVE: Adequate vitamin and trace mineral intake for pigs are important to achieve satisfactory growth performance. There are no data available on the vitamin and trace mineral intake across pig producers in China. The purpose of this study was to investigate and describe the amount of vitamin and trace minerals used in Chinese pig diets. METHODS: A 1-year survey of supplemented vitamin and trace minerals in pig diets was organized in China. A total of 69 producers were invited for the survey, which represents approximately 90% of the pig herd in China. Data were compiled by bodyweight stages to determine descriptive statistics. Nutrients were evaluated for vitamin A, vitamin D, vitamin E, vitamin K, thiamine, riboflavin, vitamin B6, vitamin B12, pantothenic acid, niacin, folic acid, biotin, choline, copper, iron, manganese, zinc, selenium, and iodine. Data were statistically analyzed by functions in Excel. RESULTS: The results indicated variation for supplemented vitamin (vitamin A, vitamin D, vitamin E, vitamin K, vitamin B12, pantothenic acid, niacin, and choline) and trace minerals (copper, manganese, zinc, and iodine) in pig diets, but most vitamins and trace minerals were included at concentrations far above the total dietary requirement estimates reported by the National Research Council and the China's Feeding Standard of Swine. CONCLUSION: The levels of vitamin and trace mineral used in China's pig industry vary widely. Adding a high concentration for vitamin and trace mineral appears to be common practice in pig diets. This investigation provides a reference for supplementation rates of the vitamins and trace minerals in the China's pig industry.

The Role of Oxymatrine in Amelioration of Acute Lung Injury Subjected to Myocardial I/R by Inhibiting Endoplasmic Reticulum Stress in Diabetic Rats.

BACKGROUND: Oxymatrine (OMT) is the primary pharmacological component of Sophora flavescens Aiton., which has been shown to possess potent antifibrotic, antioxidant, and anti-inflammatory activities. The aim of the present study was to clarify the protective mechanism of OMT on acute lung injury (ALI) subjected to myocardial ischemia/reperfusion (I/R). METHODS: A myocardial I/R-induced ALI model was achieved in diabetic rats by occluding the left anterior descending coronary artery for 1 h, followed by reperfusion for 1 h. The levels of inflammatory factors (tumor necrosis factor-α, interleukin- (IL-) 6, and IL-17) in bronchoalveolar lavage fluid were assessed using commercially available kits. The index of myocardial injury, including the detection of cardiac troponin I (cTnI), cardiac troponin T (cTnT), lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB), was also determined using commercially available kits. Hematoxylin and eosin staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling were used to identify histological changes. The expression levels of endoplasmic reticulum chaperone BiP (GRP78), DNA damage-inducible transcript 3 protein (CHOP), eukaryotic translation initiation factor 2-alpha kinase 3 (PERK), inositol dependent enzyme 1α (IRE1α), ATF6, caspase-3, -9, and-12, Bcl-2, and Bax were determined by Western blotting. The mRNA expression levels of GRP78 and CHOP were detected by reverse transcription-quantitative PCR. RESULTS: Myocardial I/R increased the levels of cTnI, cTnT, LDH, and CK-MB in diabetic rats. Damaged and irregularly arranged myocardial cells were also observed, as well as more serious ALI with higher lung injury scores and WET/DRY ratios and lower PaO2. Moreover, the expression of key proteins of endoplasmic reticulum stress (ERS) was increased by I/R injury, including phosphorylated- (p-) PERK, p-IRE1ɑ, and ATF6, as well as decreased levels of apoptosis. These effects were all significantly reversed by OMT treatment. CONCLUSIONS: OMT protects against ALI subjected to myocardial I/R by inhibiting ERS in diabetic rats.

The Attenuation of Traumatic Brain Injury via Inhibition of Oxidative Stress and Apoptosis by Tanshinone IIA.

Traumatic brain injury (TBI) is a major source of mortality and long-term disability worldwide. The mechanisms associated with TBI development are poorly understood, and little progress has been made in the treatment of TBI. Tanshinone IIA is an effective agent to treat a variety of disorders; however, the mechanisms of Tanshinone IIA on TBI remain unclear. The aim of the present study was to investigate the therapeutic potential of Tanshinone IIA on TBI and its underlying molecular mechanisms. Changes in microvascular permeability were examined to determine the extent of TBI with Evans blue dye. Brain edema was assessed by measuring the wet weight to dry weight ratio. The expression levels of CD11, interleukin- (IL-) 1ß, and tumor necrosis factor- (TNF-) α mRNA were determined by reverse transcription-quantitative PCR. Aquaporin-4 (AQP4), glial fibrillary acidic protein (GFAP), and p47phox protein expression levels were detected by western blotting. Superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-PX) activities, and malondialdehyde (MDA) content were determined using commercial kits. Cell apoptosis was detected by western blotting and TUNEL staining. Tanshinone IIA (10 mg/kg/day, intraperitoneal administration) significantly reduced brain water content and vascular permeability at 12, 24, 48, and 72 h after TBI. Tanshinone IIA downregulated the mRNA expression levels of various factors induced by TBI, including CD11, IL-1ß, and TNF-α. Notably, CD11 mRNA downregulation suggested that Tanshinone IIA inhibited microglia activation. Further results showed that Tanshinone IIA treatment significantly downregulated AQP4 and GFAP expression. TBI-induced oxidative stress and apoptosis were markedly reversed by Tanshinone IIA, with an increase in SOD and GSH-PX activities and a decrease in the MDA content. Moreover, Tanshinone IIA decreased TBI-induced NADPH oxidase activation via the inhibition of p47phox. Tanshinone IIA attenuated TBI, and its mechanism of action may involve the inhibition of oxidative stress and apoptosis.

NIR absorbing reduced graphene oxide for photothermal radiotherapy for treatment of esophageal cancer.

Theranostic agents were drawing a huge attention in the personalized medication. In this study, we established a facile technique, plant extract-based technique for the synthesis of reduced nano-graphene oxide (RNGO) with low cytotoxicity. We formed platforms of photothermal (PT) therapy and further explained that the synthesized RNGO can be utilized as ready to use PT therapy without any additional surface adjustment. In the meantime, with the help of a constant-wave NIR laser (near-infrared), in vitro esophageal adenocarcinoma cell line (OE-19) cancer cells were effectively ablated, because of the PT impact of RNGO. The outcomes propose that the RNGO was appropriate for PT therapy and photoacoustic imaging of the tumor, which is assuring for theranostic nanomedicine.

Cardioprotective Effects of Malvidin Against Isoproterenol-Induced Myocardial Infarction in Rats: A Mechanistic Study.

BACKGROUND Malvidin (alvidin-3-glucoside) is a polyphenol that belongs to the class of natural anthocyanin, which is abundantly found in red wines, colored fruits, and the skin of red grapes. Therefore, the current investigation was intended to evaluate the effect of malvidin against myocardial infarction induced by isoproterenol in the rats. MATERIAL AND METHODS The cardioprotective effects was assessed by determining the effect of malvidin on the activities of endogenous antioxidants - catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH) - and on the levels of lipid peroxidation and serum marker enzymes. The serum levels of IL-6 and TNF-α were also determined using an enzyme-linked immunosorbent assay (ELISA) kit. RESULTS The present study demonstrated a significant cardioprotective effect of malvidin by restoring the defensive activities of endogenous antioxidants - catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH) - and by reducing the levels of lipid peroxidation and serum marker enzymes lactate dehydrogenase (LD) and creatine kinase (CK). Malvidin significantly ameliorated the histopathological changes and impaired mitochondria in the cardiac necrosis stimulated with isoproterenol. Additionally, the results also demonstrated that nuclear translocation of Nrf-2 and subsequent HO-1 expression might be associated with nuclear factor kappa B (NF-κB) pathway activation. CONCLUSIONS Our findings suggest that malvidin exerts cardioprotective effects that might be due to possible strong antioxidant and anti-inflammatory activities. Therefore, this study provides the basis for the development of malvidin as a safe and effective treatment of myocardial infarction.

[Effect of Electroacupuncture at Distal and Proximal Acupoints on Expression of Spinal p 38 Mitogen-activated Protein Kinase and Cyclic Adenosine 3',5'-monophosphate in Rats with Lumbago].

OBJECTIVE: To observe the influence of Electroacupuncture (EA) stimulation of different distal-proximal acupoints combination groups on pain reaction, spinal p 38 mitogen-activated protein kinase (p 38 MAPK) expression and cyclic adenosine 3', 5'-monophosphate (cAMP) content in experimental lumbago rats. METHODS: Seventy-eight male SD rats were randomized into normal, model, sham operation, EX-B 2-BL 25 (proximal acupoints, PA), BL 40-BL 60 (distal acupoints, DA) and DA+PA groups. The lumbago model was established by implanting the autologous nucleus pulposus to the site close to the left L 5 dorsal root ganglion and the surface of dura mate of spinal cord. EA (2 Hz/100 HZ,1-3 mA) was applied at bilateral EX-B 2-BL 25 or BL 40-BL 60 or both for 20 minutes, once daily for 7 days. The mechanical pain threshold was measured, and the expression of p 38 MAPK in the lumbar spinal cord was detected by immunohistochemistry and Western blot, separately. The content of cAMP in spinal cord (L 4-L 6) was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: After mode-ling, the mechanical pain threshold percentages were significantly decreased on day 3, 5 and 7 (P<0.01), and after EA stimulation of DA,PA and DA+PA, the decreased pain threshold was significantly increased at the 3 time-points (P<0.01), and the effects of both PA and DA+PA were markedly superior to that of DA (P<0.05, P<0.01). Following modeling, lumbar spinal p 38 MAPK immunoactivity and protein expression, and cAMP content were obviously up-regulated (P<0.01). Compared with the model group, the levels of p 38 MAPK immunoactivity and protein expression in the 3 EA groups and the sham group, and cAMP contents in both PA and DA+PA groups (not in the DA and sham groups) were considerably down-regulated (P<0.01, P<0.05). CONCLUSIONS: EA stimulation of distal+proximal acupoints and proximal acupoints near the focus can raise pain threshold in lumbago rats, which may be associated with their effects in suppressing lumbago-induced increase of expression of spinal p 38 MAPK protein and cAMP content. The effects of EA of distal+proximal acupoints and proximal acupoints are better than those of EA of simple distal acupoints.

[Development of Researches on Mechanisms of Acupoint Combination for Some Disorders in Nerve-humoral-immunological Modulation].

Proper combination of acupoints is one of the core issues of acupuncturology, and is also one of the factors affecting clinical outcomes. In the present paper, the authors make a review on the development of researches on the underlying mechanisms of acupoint combination for some clinical disorders from nerve system, humoral factors and immunological regulation. The neural mechanism involves both the peripheral nervous system (mainly the segmental innervation) and central nervous system (spinal cord, brainstem, hypothalamus, cerebral cortex and cerebellum) , while the humoral factors chiefly contain neurotransmitters and hormones of the endocrine system. The immunological mechanisms involve immunocytes and cytokines. Generally speaking, following acupuncture stimulation of different combined-acupoints or acupoint groups, some synergistic effects may be produced. In fact, the relatively specific clinical effect may be easy to be obtained at the peripheral segmental innervations level.

[Chemical Constituents from Leaves of "Chuju" Chrysanthemum morifolium and Their Antioxidant Activities in vitro].

OBJECTIVE: To study the chemical constituents from the leaves of "Chuju" Chrysanthemum morifolium. METHODS: All compounds were separated and purified by column chromatography over silica gel, Sephadex LH-20 and preparative HPLC. Their structures were identified by spectral methods including 1H-NMR and 13C-NMR. RESULTS: 21 compounds were isolated and identified as octa-cosyl alcohol (1), ß-sitosterol (2), lupeol (3), α-amyrin (4), daucosterol (5), ineupatorolide B (6), syringin (7), chlorogenic acid (8), petasiphenol (9), physcion (10), acacetin (11), eupatilin (12), quercetin (13), diosmetin (14), luteolin (15), apigenin (16), apigenin- 7-O-ß-D-glucopyranoside (17), quercetin-3-O-ß-D-glucopyranoside (18), luteolin-7-O-ß-D-gluco pyranoside (19), apigenin-7-O-ß-D- neospheroside (20), and acacetin-7-O-ß-D-glucoside (21). CONCLUSION: Compounds 1-12, 18 and 20 are isolated from this plant for the first time. Compounds 10, 13, 14, 15 and 16 have shown strong antioxidant activities by DPPH · scavenging activity better than Vit C.

Administration of Lactobacillus salivarius LI01 or Pediococcus pentosaceus LI05 improves acute liver injury induced by D-galactosamine in rats.

This work investigated the effect of the intragastric administration of five lactic acid bacteria from healthy people on acute liver failure in rats. Sprague-Dawley rats were given intragastric supplements of Lactobacillus salivarius LI01, Lactobacillus salivarius LI02, Lactobacillus paracasei LI03, Lactobacillus plantarum LI04, or Pediococcus pentosaceus LI05 for 8 days. Acute liver injury was induced on the eighth day by intraperitoneal injection of 1.1 g/kg body weight D-galactosamine (D-GalN). After 24 h, samples were collected to determine the level of liver enzymes, liver function, histology of the terminal ileum and liver, serum levels of inflammatory cytokines, bacterial translocation, and composition of the gut microbiome. The results indicated that pretreatment with L. salivarius LI01 or P. pentosaceus LI05 significantly reduced elevated alanine aminotransferase and aspartate aminotransferase levels, prevented the increase in total bilirubin, reduced the histological abnormalities of both the liver and the terminal ileum, decreased bacterial translocation, increased the serum level of interleukin 10 and/or interferon-γ, and resulted in a cecal microbiome that differed from that of the liver injury control. Pretreatment with L. plantarum LI04 or L. salivarius LI02 demonstrated no significant effects during this process, and pretreatment with L. paracasei LI03 aggravated liver injury. To the best of our knowledge, the effects of the three species-L. paracasei, L. salivarius, and P. pentosaceus-on D-GalN-induced liver injury have not been previously studied. The excellent characteristics of L. salivarius LI01 and P. pentosaceus LI05 enable them to serve as potential probiotics in the prevention or treatment of acute liver failure.

Curcumin serves as a human kv1.3 blocker to inhibit effector memory T lymphocyte activities.

Curcumin, the principal active component of turmeric, has long been used to treat various diseases in India and China. Recent studies show that curcumin can serve as a therapeutic agent for autoimmune diseases via a variety of mechanisms. Effector memory T cells (T(EM), CCR7⁻ CD45RO⁺ T lymphocyte) have been demonstrated to play a crucial role in the pathogenesis of T cell-mediated autoimmune diseases, such as multiple sclerosis (MS) or rheumatoid arthritis (RA). Kv1.3 channels are predominantly expressed in T(EM) cells and control T(EM) activities. In the present study, we examined the effect of curcumin on human Kv1.3 (hKv1.3) channels stably expressed in HEK-293 cells and its ability to inhibit proliferation and cytokine secretion of T(EM) cells isolated from patients with MS or RA. Curcumin exhibited a direct blockage of hKv1.3 channels in a time-dependent and concentration-dependent manner. Moreover, the activation curve was shifted to a more positive potential, which was consistent with an open-channel blockade. Paralleling hKv1.3 inhibition, curcumin significantly inhibited proliferation and interferon-γ secretion of T(EM) cells. Our findings demonstrate that curcumin is able to inhibit proliferation and proinflammatory cytokine secretion of T(EM) cells probably through inhibition of hKv1.3 channels, which contributes to the potency of curcumin for the treatment of autoimmune diseases. This is probably one of pharmacological mechanisms of curcumin used to treat autoimmune diseases.

[Effect of tetradrine on electrophysilogic changes caused by rising of left ventricular preload in guinea pigs].

OBJECTIVE: To investigate the changes of guinea pig heart electrophysiological properties caused by increasing left ventricular preload, and to assess the effects of tetradrine on these changes. METHOD: Working model preparation of guinea pig hearts in vitro was used, and the preload of left ventricle was increased by adjusting the prefusion pressure of left atria. The changes of heart electrophysiologic parameters including monophasic action potential duration (MAPD90), monophasic action potential amplitude (MAPA), effective refractory period (ERP) and ventricular fibrillation threshold (VFT) were observed before and after altering the preload of left ventricle, and compared in the absence and presence of tetradrine, streptomycin or verapamil. RESULT: The rising of left ventricular preload led to shortening of MAPD90, ERP, and to descent of MAPA, VFT (all P<0.01). Both Tetradrine and streptomycin inhibited these changes of heart electrophysiologic parameters caused by elevation of left ventricular afterload (all P<0.01). In contrast, verapamil had no effects on the preload-related electrophysiological changes (all P>0.05). CONCLUSION: Electrophysiologic changes caused by increasing left ventricular preload may be inhibited by tetrandrine, through inhibition of stretch-activated ion channels.

Effect of tetradrine on electrophysilogic changes caused by rising of left ventricular preload in guinea pigs / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the changes of guinea pig heart electrophysiological properties caused by increasing left ventricular preload, and to assess the effects of tetradrine on these changes.</p><p><b>METHOD</b>Working model preparation of guinea pig hearts in vitro was used, and the preload of left ventricle was increased by adjusting the prefusion pressure of left atria. The changes of heart electrophysiologic parameters including monophasic action potential duration (MAPD90), monophasic action potential amplitude (MAPA), effective refractory period (ERP) and ventricular fibrillation threshold (VFT) were observed before and after altering the preload of left ventricle, and compared in the absence and presence of tetradrine, streptomycin or verapamil.</p><p><b>RESULT</b>The rising of left ventricular preload led to shortening of MAPD90, ERP, and to descent of MAPA, VFT (all P<0.01). Both Tetradrine and streptomycin inhibited these changes of heart electrophysiologic parameters caused by elevation of left ventricular afterload (all P<0.01). In contrast, verapamil had no effects on the preload-related electrophysiological changes (all P>0.05).</p><p><b>CONCLUSION</b>Electrophysiologic changes caused by increasing left ventricular preload may be inhibited by tetrandrine, through inhibition of stretch-activated ion channels.</p>