TMT and PRM Based Quantitative Proteomics to Explore the Protective Role and Mechanism of Iristectorin B in Stroke.

Stroke is a serious disease caused by the rupture or blockage of the cerebrovascular system. Its pathogenesis is complex and involves multiple mechanisms. Iristectorin B is a natural isoflavone that has certain anti stroke effects. In this study, an in vitro stroke injury model of glyoxylate deprivation was established using PC12 cells, which was used to evaluate the anti-stroke activity of Iristectorin B in ejecta stem. The results showed that Iristectorin B, a natural isoflavone derived from Dried Shoot, significantly reduced the damage to PC12 cells caused by oxygen glucose deprivation/reoxygenation, decreased apoptosis, enhanced cell survival and reduced Ca2+, LDH and ROS levels. The results showed that Iristectorin B had a significant protective effect on Na2S2O4-injured PC12 cells, and the mechanism may be related to the protective effect of neurons in the brain. After protein extraction and various analyses were performed, a series of cutting-edge technologies were organically combined to study the quantitative proteome of each group. Differential proteins were then analyzed. According to the protein screening principle, ferroptosis-related proteins were most closely associated with stroke. The differential proteins associated with ferroptosis screened were SLC3A2, TFR1 and HMOX1, with HMOX1 being the most significantly elevated and reduced via dosing. Iristectorin B may act as a protective agent against stroke by regulating ferroptosis, and SLC3A2, TFR1 and HMOX1 may serve as potential diagnostic biomarkers for stroke, providing additional evidence to support the importance of ferroptosis in stroke.

The protection impact of tectoridin on PC12 cell preventing OGD/R-caused damage through PI3K/AKT signaling channel.

The present work examined the effect exerted by tectoridin preventing oxygen glucose deprivation/reoxygenation (OGD/R) damage within PC12 cell. We incubated PC12 cells with Na2S2O4 (10 mM) for 2 h, and tectoridin at different concentrations was then added; based on methyl-thiazolyl-tetrazolium (MTT) and lactate dehydrogenase (LDH) tests, the protection impact was tested. 2',7'-dicholorofluorescein diacetate (DCFH-DA), Fluo-3AM, and 5, 5', 6, 6' -tetrachloro-1, 1', 3, 3' -tetraethyl-imidacarbocyanine iodide (JC-1) staining, and Western blotting were used for determining reactive oxygen species (ROS) level, intracellular Ca2+ content, mitochondrial membrane potential (MMP) and the related proteins contents. As a result, tectoridin could improve the cell viability and inhibit the release of LDH. In-depth studies demonstrated that tectoridin limited the overproduction of ROS and intracellular Ca2+ content and increased MMP, which showed a close association with ROS-mediated mitochondrial function. Moreover, tectoridin hindered apoptosis based on the up-regulation of the expressions of p-AKT, Bcl-2/Bax and p-mTOR. Furthermore, the level of Nrf2 was also improved by treatment of tectoridin. In addition, the expression of Bcl-2/Bax, p-Akt, p-mTOR, Nrf2, HO-1, NQO1 and GCLM were reduced by LY294002 and the protective role of tectoridin was limited by LY294002. The results unambiguously suggested that tectoridin reduced OGD/R-caused damage to PC12 cells and might ensure neuroprotection by stimulating the PI3K/AKT signaling channel.

Efficacy of Epigallocatechin-3-Gallate in Preventing Dermatitis in Patients With Breast Cancer Receiving Postoperative Radiotherapy: A Double-Blind, Placebo-Controlled, Phase 2 Randomized Clinical Trial.

Importance: Safe and effective prophylactic therapies for radiation-induced dermatitis (RID) remain an unmet need. Objective: To determine if epigallocatechin-3-gallate (EGCG) solution reduces the incidence of RID in patients undergoing radiotherapy after breast cancer surgery. Design, Setting, and Participants: This phase 2 double-blind, placebo-controlled randomized clinical trial enrolled 180 patients with breast cancer receiving postoperative radiotherapy at Shandong Cancer Hospital and Institute in Shandong, China, between November 2014 and June 2019. Data analysis was performed from September 2019 to January 2020. Interventions: Participants were randomly assigned (2:1) to receive either EGCG solution (660 µmol/L) or placebo (0.9% NaCl saline) sprayed to the whole radiation field from day 1 of the radiation until 2 weeks after radiation completion. Main Outcomes and Measures: The primary end point was incidence of grade 2 or worse RID, defined by the Radiation Therapy Oncology Group scale. The secondary end points included RID index (RIDI), symptom index, changes in the skin temperature measured by infrared thermal images, and safety. Results: A total of 180 eligible patients were enrolled, of whom 165 (EGCG, n = 111; placebo, n = 54) were evaluable for efficacy (median [range] age, 46 [26-67] years). The occurrence of grade 2 or worse RID was significantly lower (50.5%; 95% CI, 41.2%-59.8%) in the EGCG group than in the placebo group (72.2%; 95% CI, 60.3%-84.1%) (P = .008). The mean RIDI in the EGCG group was significantly lower than that in the placebo group. Furthermore, symptom indexes were significantly lower in patients receiving EGCG. Four patients (3.6%) had adverse events related to the EGCG treatment, including grade 1 pricking skin sensation (3 [2.7%]) and pruritus (1 [0.9%]). Conclusions and Relevance: In this randomized clinical trial, prophylactic use of EGCG solution significantly reduced the incidence and severity of RID in patients receiving adjuvant radiotherapy for breast cancer. It has the potential to become a new choice of skin care for patients receiving radiotherapy. Trial Registration: ClinicalTrials.gov Identifier: NCT02580279.

Screening anti-gout compounds from Phellinus igniarius by ultrafiltration liquid chromatography mass spectrometry based on evaluation of an in vitro method combined with enzymatic reaction.

In the present study, the anti-inflammation effect of Phellinus igniarius extract was detected on an in vitro model of RAW 264.7 cells stimulated using sodium urate. In this cell model, the content changes of inflammatory cytokines, intercellular adhesion molecule-1, and interleukin-1 beta, in cell culture supernatants were detected using an enzyme-linked immunosorbent assay. The xanthine oxidase inhibitory activity of P. igniarius extracts was determined using a microplate reader. Furthermore, in order to identify the active compounds of P. igniarius, ultrafiltration liquid chromatography mass spectrometry was utilized to screen xanthine oxidase inhibitors from the extract. Our results showed that in the presence of P. igniarius extract, the expressions of interleukin-1 beta and intercellular adhesion molecule-1 decreased (p < 0.01 and p < 0.05, respectively) compared to that in the control group. The extract effective inhibited the xanthine oxidase activity. Finally, seven compounds were screened and identified as potential xanthine oxidase inhibitors from P. igniarius. Taken together, these results demonstrate a potential anti-inflammation bioactivity of P. igniarius in vitro, providing a basis for further in vivo research for the prevention and treatment of gout.

Dual-energy computed tomography could reliably differentiate metastatic from non-metastatic lymph nodes of less than 0.5 cm in patients with papillary thyroid carcinoma.

BACKGROUND: Dual-energy computed tomography (DECT) has been widely applied to detect lymph node (LN) and lymph node metastasis (LNM) in various cancers, including papillary thyroid carcinoma (PTC). This study aimed to quantitatively evaluate metastatic cervical lymph nodes (LNs) <0.5 cm in patients with PTC using DECT, which has not been done in previous studies. METHODS: Preoperative DECT data of patients with pathologically confirmed PTC were retrospectively collected and analyzed between May 2016 and June 2018. A total of 359 LNs from 52 patients were included. Diameter, iodine concentration (IC), normalized iodine concentration (NIC), and the slope of the energy spectrum curve (λHU) of LNs in the arterial and the venous phases were compared between metastatic and non-metastatic LNs. The optimal parameters were obtained from the receiver operating characteristic (ROC) curves. The generalized estimation equation (GEE) model was used to evaluate independent diagnostic factors for LNM. RESULTS: A total of 139 metastatic and 220 non-metastatic LNs were analyzed. There were statistical differences of quantitative parameters between the two groups (P value 0.000-0.007). The optimal parameter for diagnosing LNM was IC in the arterial phase, and its area under the curve (AUC), sensitivity, and specificity were 0.775, 71.9%, and 73.6%, respectively. When the three parameters of diameter, IC in the arterial phase, and NIC in the venous phase were combined, the prediction efficiency was better, and the AUC was 0.819. The GEE results showed that LNs located in level VIa [odds ratio (OR) 2.030, 95% confidence interval (CI): 1.134-3.634, P=0.017], VIb (OR 2.836, 95% CI: 1.597-5.038, P=0.000), diameter (OR 2.023, 95% CI: 1.158-3.532, P=0.013), IC in the arterial phase (OR 4.444, 95% CI: 2.808-7.035, P=0.000), and IC in the venous phase (OR 5.387, 95% CI: 3.449-8.413, P=0.000) were independent risk factors for LNM in patients with PTC. CONCLUSIONS: DECT had good diagnostic performance in the differentiation of cervical metastatic LNs <0.5 cm in patients with PTC.

CircHIF1A regulated by FUS accelerates triple-negative breast cancer progression by modulating NFIB expression and translocation.

Emerging evidence has demonstrated that circular RNAs (circRNAs) play critical roles in the development and progression of human cancer. However, the biological functions and underlying mechanisms of circRNAs in triple-negative breast cancer (TNBC) remain to be investigated. In our present study, we found that the novel circRNA circHIF1A was significantly overexpressed in breast cancer tissues and that it was associated with metastasis, poor prognosis, and the TNBC subtype. Gain- and loss-of-function experiments were conducted to investigate the biological roles of circHIF1A in TNBC. Overexpression of circHIF1A significantly promoted TNBC growth and metastasis in vitro and in vivo, while knockdown of circHIF1A exerted the opposite effects. Mechanistically, circHIF1A modulated the expression and translocation of NFIB through posttranscriptional and posttranslational modifications, resulting in the activation of the AKT/STAT3 signaling pathway and inhibition of P21. The RNA binding protein FUS could regulate the biogenesis of circHIF1A by interacting with the flanking intron, and FUS was transcriptionally regulated by NFIB, thus forming the circHIF1A/NFIB/FUS positive feedback loop. Moreover, circHIF1A could be packaged into exosomes and was upregulated in the plasma of breast cancer patients. Our findings indicated that circHIF1A played a critical role in the growth and metastasis of TNBC via a positive feedback loop and that circHIF1A could be a promising biomarker for breast cancer diagnosis and a potential therapeutic target for TNBC treatment.

Evaluation of bone marrow infiltration in multiple myeloma using whole-body diffusion-weighted imaging and T1-weighted water-fat separation Dixon.

BACKGROUND: Multiple myeloma (MM) is a blood cancer caused by the unlimited proliferation of intramedullary plasma cells. The presence of focal lesions (FLs) is presumed to be a more relevant factor for patient outcomes and risk distribution than diffuse bone marrow signal abnormalities. Signal changes in these FLs also have a good correlation with prognosis. As the cell density increased, a lower apparent diffusion coefficient (ADC) value was found with the diffusion-weighted imaging (DWI) sequence. Therefore, whole-body magnetic resonance imaging (MRI) with DWI sequences is sensitive to cell density and viability and may be vital for disease detection and therapy response assessments. However, the correlation between the DWI signal and the degree of bone destruction and the proportion of bone marrow plasma cells (BMPC) was still unclear in patients with MM. Water-fat separation MRI is used mainly for evaluating liver and bone marrow fat quantification, and fat quantification in other diseases. Meanwhile, it is also possible to assess the extent of bone marrow invasion in medullary lesions. This study aimed to investigate the correlation between ADC values from whole-body DWI and water/fat MRI signals from T1-weighted water-fat separation in evaluating bone marrow infiltration in patients with MM. METHODS: The study included 35 patients with MM who underwent whole-body DWI and T1-weighted water-fat separation Dixon examinations before therapy. The ADC values, normalized fat signal intensity (nMfat), normalized water molecular signal intensity (nMwater), and normalized fat fraction (nFF) of the thoracolumbar spine was measured in FLs and the normal-appearing bone marrow (NABM). The differences in values were compared using the independent-samples t-test. The correlation between ADC values and water-fat MRI signals was estimated using the Pearson or Spearman correlation test. The correlation between the MRI above parameters and proportions of BMPC was also explored. RESULTS: Statistically significant differences were found between the mean ADC values in FLs and NABM (0.72 vs. 0.33 mm2/s, P<0.0001). Significantly elevated nMwater values and decreased nMfat and nFF values were observed in FLs; no correlations were found in NABM (P>0.05). The ADC value highly correlated with nMfat and nFF values and moderately with the nMwater value in FLs (r=-0.899, -0.834, 0.642, respectively, P<0.0001). Correlations were also observed between the proportion of BMPC and MRI parameters in MM (r=0.984, 0.716, -0.938, and -0.905, respectively, P<0.05). CONCLUSIONS: The ADC value combined with water-fat separation parameters could be used for evaluating thoracolumbar bone marrow infiltration in MM. All parameters correlated with the proportion of BMPC, which helped assess the early response in MM therapy.

Quantitative susceptibility-weighted imaging may be an accurate method for determining stroke hypoperfusion and hypoxia of penumbra.

OBJECTIVES: To quantitatively evaluate the volume of the ischemic penumbra using susceptibility-weighted imaging and mapping (SWIM) of asymmetrical prominent cortical veins (APCVs) in patients with acute ischemic stroke. METHODS: Eighty-five eligible patients with acute ischemic stroke on admission within 12 h from symptom onset were studied. The APCVs on SWIM were quantitatively (SWI-volume) and semi-quantitatively (SWI-Alberta Stroke Program Early CT Score, SWI-ASPECTS) evaluated to calculate mismatch. To assess the diagnostic efficacy of APCVs on SWIM, comparative analyses were performed between SWIvolume-DWI mismatch and SWIASPECTS-DWI mismatch, using PWI-DWI mismatch as a reference. Correlations were calculated between the mismatches, as well as between SWI-volume and time-to-maximum (Tmax) > 6 s volume. Additionally, each of these mismatches was correlated with the National Institute of Health Stroke Scale (NIHSS). RESULTS: The sensitivity, negative predictive value, and accuracy of SWIvolume-DWI mismatch were demonstrably higher than SWIASPECTS-DWI mismatch (100% vs. 53.7%, 100% vs. 9.5%, 97.7% vs. 54.5%, respectively). A significant positive correlation was found between SWIvolume-DWI and PWI-DWI mismatch (r = 0.691, p < 0.01), as well as between SWI-volume and Tmax > 6 s volume (r = 0.786, p < 0.001). A significant negative correlation was found between SWIvolume-DWI mismatch and NIHSS (r = - 0.360, p = 0.022), as well as between SWIASPECTS-DWI mismatch and NIHSS (r = - 0.499, p = 0.001). CONCLUSIONS: SWIvolume-DWI mismatch had higher diagnostic efficacy than SWIASPECTS-DWI mismatch in defining the ischemic penumbra and showed good consistency with PWI-DWI mismatch in acute ischemic stroke. Quantitation of APCVs using SWIM provided an accurate method for determining hypoperfusion and provided a reliable method to reflect the hypoxia of penumbra. KEY POINTS: • SWIvolume-DWI mismatch has higher diagnostic efficacy than SWIASPECTS-DWI mismatch in defining the ischemic penumbra. • SWIvolume-DWI mismatch shows good consistency with PWI-DWI mismatch in managing penumbra in acute ischemic stroke. • Quantitation of APCV volume using SWIM provided an accurate method for determining the hypoperfusion area and provided a reliable method to reflect the hypoxia of penumbra.

Neuroprotective Effect of Daidzein Extracted From Pueraria lobate Radix in a Stroke Model Via the Akt/mTOR/BDNF Channel.

Daidzein is a plant isoflavonoid primarily isolated from Pueraria lobate Radix as the dry root of P. lobata (Wild.) Ohwi, have long been used as nutraceutical and medicinal herb in China. Despite the report that daidzein can prevent neuronal damage and improve outcome in experimental stroke, the mechanisms of this neuroprotective action have been not fully elucidated. The aim of this study was to determine whether the daidzein elicits beneficial actions in a stroke model, namely, cerebral ischemia/reperfusion (I/R) injury, and to reveal the underlying neuroprotective mechanisms associated with the regulation of Akt/mTOR/BDNF signal pathway. The results showed that I/R, daidzein treatment significantly improved neurological deficits, infarct volume, and brain edema at 20 and 30 mg/kg, respectively. Meanwhile, it was found out that the pretreatment with daidzein at 20 and 30 mg/kg evidently improved striatal dopamine and its metabolite levels. In addition, daidzein treatment reduced the cleaved Caspase-3 level but enhanced the phosphorylation of Akt, BAD and mTOR. Moreover, daidzein at 30 mg/kg treatment enhanced the expression of BDNF and CREB significantly. This protective effect of daidzein was ameliorated by inhibiting the PI3K/Akt/mTOR signaling pathway using LY294002. To sum up, our results demonstrated that daidzein could protect animals against ischemic damage through the regulation of the Akt/mTOR/BDNF channel, and the present study may facilitate the therapeutic research of stroke.

Alkaloids extracted from Uncaria rhynchophylla demonstrate neuroprotective effects in MPTP-induced experimental parkinsonism by regulating the PI3K/Akt/mTOR signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Alkaloids isolated from Uncaria rhynchophylla (Miq.) Miq. ex Havil. (Rubiaceae), alkaloids (URA) have been used to treat diseases related to the central nervous system, such as Parkinson's disease. Nevertheless, the potential mechanisms underlying their neuroprotective effects are not well-understood. AIM OF THE STUDY: We investigated the neuroprotective effects of URAs in a mouse model of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) and the possible involvement of a molecular signaling pathway. MATERIALS AND METHODS: Two typical experiments for animal behavior despair, the spontaneous motor activity and the rotarod experiments, were employed to evaluate the efficacy of URAs in mice with PD symptoms. Dopamine (DA) neurons and their metabolism were evaluated using high-performance liquid chromatography-tandem mass spectrometry. The mechanism of action of the alkaloids was investigated by analyzing their effects on the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway using western blotting. RESULTS: URA treatment effectively improved the behaviors of the mice during the "spontaneous motor activity and latency to fall off the rotarod test". Moreover, URAs demonstrated a protective role in dopaminergic neurons by increasing the expression of the dopamine transporter and tyrosine hydroxylase, which were supposed to be reduced by MPTP, inhibiting dopamine turnover, and changing dopamine and relevant metabolites. In addition to its association with the increase in the Bcl-2/Bad ratio, URA treatment also attenuated the cleaved caspase-3 level and enhanced the phosphorylation of Akt and mTOR. CONCLUSION: These findings provide evidence that URA can effectively protect neurons from the neurotoxicity caused by MPTP in mouse models of PD by up-regulating the PI3K/Akt/mTOR signaling pathway.

Plaque characteristics and hemodynamics contribute to neurological impairment in patients with ischemic stroke and transient ischemic attack.

OBJECTIVES: We aimed to investigate differential characteristics of plaque in the middle cerebral artery (MCA) and hemodynamics in patients with ischemic stroke and transient ischemic attack (TIA), and to develop a predictive model for the presence of ischemic stroke and neurological impairment. METHODS: Sixty-seven patients with acute ischemic events in MCA territory who underwent high-resolution vessel wall imaging between September 2016 and August 2018 were reviewed retrospectively. Patients were assigned to either the stroke group or TIA group, according to diffusion-weighted imaging and neurological examination. Plaque characteristics and anterograde score (AnS) were calculated. Tmax > 6.0-s volume was acquired by RApid Processing of perfusIon and Diffusion software. Multivariate logistic regression analysis and multiple linear regression analysis were performed to establish a predictive model for irreversible infarction occurrence and clinical severity. RESULTS: Forty-five patients were assigned to the stroke group, and 22 were assigned to the TIA group. Plaque length, intraplaque hemorrhage (IPH), enhancement, AnS, and Tmax > 6.0-s volumes were significantly different between the two groups (p < 0.05). IPH and AnS were independent predictors for patients with stroke (p = 0.020 and 0.034, respectively). Tmax > 6.0-s volume, IPH, hypertension, and AnS were associated with high National Institutes of Health Stroke Scale (NIHSS) scores (all p < 0.05, R = 0.725, and adjusted R2 = 0.494). CONCLUSIONS: IPH and AnS are useful in predicting stroke occurrence. Tmax > 6.0-s volume, IPH, hypertension, and AnS are associated with neurological impairment of the patients. KEY POINTS: • Ischemic stroke and TIA patients have different plaque characteristics and hemodynamics. • Intraplaque hemorrhage and anterograde score have high diagnostic efficiency for ischemic stroke. • The combination of Tmax > 6.0-s volume, intraplaque hemorrhage, hypertension, and anterograde score can predict the National Institutes of Health Stroke Scale scores of patients.

Protection by rhynchophylline against MPTP/MPP+-induced neurotoxicity via regulating PI3K/Akt pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Isolated from Uncaria rhynchophylla (U. rhynchophylla), rhynchophylline (Rhy) has been applied for treating diseases related to central nervous system such as Parkinson's disease. Nevertheless, the molecular mechanism of the neuroprotective effect has not been well interpreted. AIM OF THE STUDY: To investigate the effects of Rhy on MPTP/MPP + -induced neurotoxicity in C57BL/6 mice or PC12 cells and study the mechanisms involved. MATERIALS AND METHODS: The neuroprotective effect of Rhy on MPTP-induced neurotoxicity was evaluated by spontaneous motor activity test, as well as a test of rota-rod on a rat model of Parkinson's disease. The numbers of TH-positive neurons in the substantia nigra pars compacta (SNpc) was assessed by immunohistological. CCK-8, lactate dehydrogenase (LDH), reactive oxygen species (ROS), the concentration of intracellular calcium ([Ca2+]i) and flow cytometry analysis were performed to evaluate the pharmacological property of Rhy on 1-methyl-4-phenylpyridinium (MPP+) induced neurotoxicity in PC12 cells. Besides, LY294002, a PI3K inhibitor was employed to determine the underlying molecular signaling pathway revealing the effect of Rhy by western-blot analysis. RESULTS: The results showed that Rhy exhibited a protective effect against the MPTP-induced decrease in tyrosine hydroxylase (TH)-positive fibers in the substantia nigra at 30 mg/kg, demonstrated by the immunohistological and behavioral outcomes. Furthermore, it has been indicated that cell viability was improved and the MPP+-induced apoptosis was inhibited after the treatment of Rhy at 20 µM, which were severally analyzed by the CCK-8 and the Annexin V/propidium iodide staining method. In addition, Rhy treatment attenuated MPP+-induced up-regulation of LDH, ([Ca2+]i), and the levels of ROS. Besides, it can be revealed from the Western blot assay that LY294002, as a selective Phosphatidylinositol 3-Kinase (PI3K) inhibitor, effectively inhibited the Akt phosphorylation caused by Rhy, which suggested that Rhy showed its protective property through the activated the PI3K/Akt signaling pathway. Moreover, the Rhy-induced decreases of Bax and caspase-3 as the proapoptotic markers and the increase of Bcl-2 as the antiapoptotic marker, were blocked by LY294002 in the MPP+-treated PC12 cells. CONCLUSIONS: Rhy exerts a neuroprotective effect is partly mediated by activating the PI3K/Akt signaling pathway.

Determination and Pharmacokinetics of Okanin in Rat Plasma by UltraHigh Performance Liquid Chromatography Coupled with Triple-Quadrupole Tandem Mass Spectrometry.

Okanin is a major flavonoid found in Coreopsis tinctoria Nutt., arousing huge interest recently for its considerable biological characteristics including antioxidant, antineurotoxic, and antidiabetic activities. An ultrahigh performance liquid chromatography triple-quadrupole tandem mass spectrometry (UPLC-MS) was successfully used to determine okanin in rat plasma after oral administration of okanin. Bavachalcone acted as an internal standard (IS). By gradient elution, IS and analyte were separated on a C18 column for 7 min at a flow rate of 0.25 mL/min with acetonitrile-0.1% acetic acid mobile phase. The stability, matrix effect, extraction recovery, accuracy, precision, linearity, and selectivity of the method were firstly demonstrated. The major pharmacokinetic parameters of okanin in rat plasma were then measured using the developed UPLC-MS method. An UPLC-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was finally established to obtain the specific and accurate mass of okanin in rat plasma after oral administration, and its proposed fragmentation was further elaborated.

Rapid screening and evaluation of XOD inhibitors and O2 •- scavenger from total flavonoids of Ginkgo biloba leaves by LC-MS and multimode microplate reader.

Superoxide anion radical scavenger and xanthine oxidase inhibitor play an important role in the treatment of several relevant human diseases. In the present study, ultrafiltration liquid chromatography-mass spectrometry coupled to microplate reader was applied to screen and identify superoxide anion radical scavengers and xanthine oxidase inhibitors from total flavonoids of Ginkgo biloba leaves. As a result, four compounds (quercetin, apigenin, kaempferol and isorhamnetin) were screened as xanthine oxidase inhibitors by ultrafiltration LC-MS, and the 50% scavenging concentration values of the screened flavonoids were lower than those for allopurinol. Lineweaver-Burk plot results indicated that kaempferol was a competitive xanthine oxidase inhibitor; the other flavonoids were all anticompetitive inhibitors. Four flavonoids-rutin, quercetin, kaempferol and isorhamnetin-were screened as superoxide anion radical scavengers by LC-MS. The results demonstrate that the method for screening and evaluation of superoxide anion radical scavenger and xanthine oxidase inhibitor from a complex mixture system is feasible and efficient.

Sec-O-glucosylhamaudol suppressed inflammatory reaction induced by LPS in RAW264.7 cells through inhibition of NF-κB and MAPKs signaling.

As a major bioactive compound from the Saposhnikovia divaricata (Turcz.) Schischk, sec-O-glucosylhamaudol (SOG), has been reported to have anti-nociceptive activity and high 5-lipoxygenase (5-LOX) activity. Nevertheless, the mechanism of the potential anti-inflammatory effects of SOG is unclear. The anti-inflammatory impacts of SOG in RAW 264.7 cell lines stimulated by LPS were explored in the present study. It was found that SOG dose-dependently reduced the emergence of inflammation cytokines, such as IL-6 and TNF-α in Raw264.7 murine macrophages stimulated by LPS. Real-time PCR assay demonstrated the SOG dose-dependently inhibited transcription of these cytokines as well. In addition, it was also found that NF-κB activation and MAPKs phosphorylation including p38, JNK and ERK1/2 induced by LPS were suppressed by SOG. Due to its anti-inflammatory activity, our results suggest that SOG might have therapeutic effects on inflammatory disease, such as acute lung injury or rheumatoid arthritis.

Middle cerebral artery thrombus susceptibility-weighted imaging mapping predicts prognosis.

BACKGROUND: Susceptibility weighted imaging and mapping (SWIM) of magnetic resonance imaging (MRI) is used to evaluate cerebral arterial thrombosis. The aim of this research was to assess susceptibility, length, and clot burden score (CBS) of thrombus in the middle cerebral artery (MCA) and their relationship with cerebral infarction and early clinical prognosis in patients with acute or subacute cerebral infarction. METHODS: In total, 56 patients with acute or subacute cerebral infarction (with the time from onset to admission less than 72 h) and only unilateral MCA occlusion were included in the current study. All the patients had the corresponding susceptibility vessel sign (SVS) on susceptibility-weighted imaging (SWI). Parameters including susceptibility, length, and CBS of thrombus were obtained from SWI and SWIM. The differences in susceptibility of different portions of the thrombus were compared with each other by one-way ANOVA test. The relationship between susceptibility and stroke onset time was further analyzed by Spearman correlation analysis, in addition to the relationships between susceptibility, length, CBS, diffusion-weighted imaging-Alberta stroke program early CT score (DWI-ASPECTS), and admission and discharge National Institutes of Health Stroke Scale (NIHSS). RESULTS: The susceptibility among different portions and different segments of thrombus showed no statistical difference. The susceptibility and length were weakly yet negatively correlated with DWI-ASPECTS (rs=-0.382, -0.457; P=0.004, 0.000). The susceptibility was weakly yet positively correlated with admission NIHSS and discharged NIHSS (rs=0.403, 0.430; P=0.002, 0.001). CBS was weakly yet positively correlated with DWI-ASPECTS (rs=0.349; P=0.008) and weakly yet negatively correlated with admission and discharged NIHSS (rs=-0.375, -0.335; P=0.004, 0.012). CONCLUSIONS: The susceptibility remained consistent regardless of location, length, and onset time, which indicates that the thrombus composition was similar when detected on SWI less than 72 h from the onset. Susceptibility and CBS may help to predict clinical severity and short-term clinical prognosis to some extent.

Total glucosides of paeony (TGP) extracted from Radix Paeoniae Alba exerts neuroprotective effects in MPTP-induced experimental parkinsonism by regulating the cAMP/PKA/CREB signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The totally-amounted glucosides of paeony (TGP), which are made up of paeoniflorin, albiflorin, oxypaeoniflorin as well as benzoylpaeoniflorin, constitute the Baishao' actively-working component extracted from Radix Paeonia alba employed in conventional oriental medicine aiming to treat cerebrovascular disorders, such as Parkinson's disease. However, its pharmacologic mechanism is not clear. AIM OF THE STUDY: The initial investigation was made on TGP's neuroprotective effects on PD of the mouse model based on 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) as well as the identification of potential involvement of a molecular signaling pathway. MATERIALS AND METHODS: The evaluation of the behavioral damage as well as neurotoxicity in mice was made through MPTP. Spontaneous motor activity test, as well as a test of Rota-rod on mice was employed for the measurement of bradykinesia symptom. Additionally, liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) works as the determiner of the main monoamine neurotransmitters dopamine (DA) along with its metabolites 3, 4-dihydroxyphenylacetic acid (DOPAC) as well as homovanillic acid (HVA) based on mouse hippocampus connected with the anti-Parkinson's disease like effect of TGP. Besides, the measurement of the effects of TGP treatment on the expressions level of TH, DAT, a-synuclein, p-CREBS133 as well as apoptosis influence was made with the help of western-blot assay with apoptosis-related markers such as Bax and Bcl-2. RESULTS: The results showed that TGP treatment lessened the behavior-based loss shown "in the spontaneous motor activity as well as the potential of falling to rotarod test". In addition, we found that pretreatment with TGP markedly improved motor coordination, striatal dopamine and its metabolite levels. Furthermore, pretreatment of TGP conducted the protection for dopaminergic neurons with the prevented MPTP-induced reductions within the tyrosine hydroxylase (TH), substantia nigra dopaminergic transporter (DAT), as well as increasing α-synuclein protein levels with transformed dopamine catabolism as well as inhibited dopamine turnover. Besides, TGP treatment helped reversed apoptosis signaling molecules Bcl-2/Bax' reduction; meanwhile improving p-CREBS133 the factor of growth signaling in the substantia nigra' decrease. CONCLUSION: These results suggested that TGP can enhance dopaminergic neuron's cell survival in the SNpc in virtue of the activated cAMP/PKA/CREB factor of growth on inhibiting the pathway of second messenger apoptosis as well. In conclusion, the current findings indicate TGP is expected to be a new cure for PD.

Simultaneous ultra-performance liquid chromatography-tandem mass spectrometry determination of six components in rat plasma after oral administration of Smilacis glabrae Roxb. extract.

In this study, an accurate and reliable method of ultra-performance liquid chromatography coupled with a triple-quadrupole tandem mass spectrometry was firstly developed and fully validated for the simultaneous determination of epicatechin, neoastilbin, astilbin, isoastilbin, engeletin and resveratrol in rat plasma after administration of Smilacis glabrae Roxb. extract. Naringenin was used as an internal standard (IS). The analyte and IS were separated on a C18 column by gradient elution with a mobile phase of acetonitrile-0.3% acetic acid at a flow rate of 0.25 mL/min for a total run time of 8 min. The method was validated in terms of selectivity, linearity, precision, accuracy, extraction recovery, matrix effect and stability. The developed method was successfully applied to determine the main pharmacokinetic parameters of six components in rat plasma.

Improving breast cancer therapy using doxorubicin loaded solid lipid nanoparticles: Synthesis of a novel arginine-glycine-aspartic tripeptide conjugated, pH sensitive lipid and evaluation of the nanomedicine in vitro and in vivo.

Breast cancer is the leading cause of cancer mortality in women worldwide. To overcome the toxic side effects and multidrug resistance (MDR) during doxorubicin (DOX) chemotherapy, an arginine-glycine-aspartic (RGD) tripeptide modified, pH-sensitive solid lipid nanoparticles (SLNs) is employed in this study. In this study, a RGD conjugated, pH sensitive lipid was synthesized using glycerin monostearate (GMS) and adipic acid dihydrazide (HZ) as lipid materials and named RGD-HZ-GMS. RGD-HZ-GMS was applied to encapsulate DOX to construct a RGD modified, DOX loaded SLNs (RGD-DOX-SLNs). To evaluate the anticancer effect of RGD-DOX-SLNs, breast cancer cell line (MCF-7 cells) and DOX resistant cell line (MCF-7/ADR cells) were used. in vivo tumor suspension and toxicity effects were evaluated on mice bearing MCF-7/ADR cells breast cancer model. RGD-DOX-SLNs had a uniformly spherical shape. The mean particle size and zeta potential of the RGD-DOX-SLNs was 96.3 nm and 35.6 mV, respectively. RGD-DOX-SLNs showed 5.58 fold higher area under the plasma concentration - time curve (AUC) compared with DOX solution. Terminal half life (T1/2) and peak concentration (Cmax) of RGD-DOX-SLNs was 10.85 h and 39.12 ± 2.71 L/kg/h. in vitro and in vivo antitumor results indicate that RGD-DOX-SLNs might be a promising novel lipid carrier which could improve breast cancer therapy.

Comparison and Correlation of Magnetic Resonance Imaging and Clinical Severity in Nonhuman Immunodeficiency Virus Patients with Cryptococcal Infection of Central Nervous System.

BACKGROUND: The incidence of cryptococcal meningitis among immunocompetent patients increases, especially in China and imaging plays an important role. The current study was to find the correlation between magnetic resonance imaging (MRI) manifestation and clinical severity in nonhuman immunodeficiency virus patients with cryptococcal infection of central nervous system (CNS). METHODS: A total of 65 patients with CNS cryptococcal infection from August 2014 to October 2016 were retrospectively included in this study. All the patients had MRI data and clinical data. The patients were divided into two groups according to whether the patients were confirmed with identifiable underlying disease. Comparison and correlation of MRI and clinical data in both groups were investigated using independent sample t- test, Chi-square test, Mann-Whitney test and Spearman rank correlation analysis. RESULTS: In all 65 patients, 41 cases (41/65, 63.1%; Group 1) had normal immunity and 24 cases (24/65, 36.9%; Group 2) had at least one identifiable underlying disease. Fever, higher percentage of neutrophil (NEUT) in white blood cell (WBC), and increased cell number of cerebral spinal fluid (CSF) were much common in patients with underlying disease (Group 1 vs. Group 2: Fever: 21/41 vs. 21/24, χ2 = 8.715, P = 0.003; NEUT in WBC: 73.15% vs. 79.60%, Z = -2.370, P = 0.018; cell number of CSF: 19 vs. 200, Z = -4.298, P < 0.001; respectively). Compared to the patients with normal immunity, the lesions are more common in the basal ganglia among patients with identifiable underlying disease (Group 1 vs. Group 2: 20/41 vs. 20/24, χ2 = 7.636, P = 0.006). The number of the involved brain areas in patients with identifiable underlying disease were well correlated with the number of cells and pressure of CSF (r = -0.472, P = 0.031; r = 0.779, P = 0.039; respectively). CONCLUSIONS: With the increased number of the involved brain areas in patients with identifiable underlying disease, the body has lower immunity against the organism which might result in higher intracranial pressure and more severe clinical status.