Effect of miltirone on proliferation and apoptosis of leukemia THP-1 cells / 中国中药杂志

To investigate the toxicity and related mechanism of miltirone to human acute myeloid leukemia THP-1 cells. To be specific, the active components and targets of miltirone were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the target proteins were converted into standard gene names with UniProt. Acute leukemia-rela-ted target genes were screened from GeneCards and DisGeNET. Venn diagram was constructed with Venny 2.1 to yield the common targets of the disease and the drug. The protein-protein interaction(PPI) network was constructed by STRING and Cytoscape 3.8.2. THP-1 cells in the logarithmic growth phase were treated with dimethyl sulfoxide(DMSO), and 2.5, 5, 10, 15, and 20 μmol·L~(-1) miltirone for 24 h, respectively. The proliferation rate of cells was analyzed by carboxyfluorescein diacetate succinimidyl ester(CFSE), apoptosis rate by flow cytometry with Annexin V-PE/7 AAD staining, and cell morphology by acridine orange staining. Real-time quantitative PCR(qPCR) was employed to detect the mRNA levels of nuclear receptor coactivator 2(NCOA2), poly(ADP-ribose) polymerase-1(PARP1), B-cell lymphoma-2(Bcl-2)-associated X protein(Bax), Bcl-2, and cysteine aspartyl protease-3(caspase-3). The effect of miltirone on apoptosis was detected in presence of caspase inhibitor Z-VAD-FMK. A total of 26 targets of miltirone, 1 046 genes related to acute leukemia, and 6 common targets of the two were screened out. Flow cytometry result showed miltirone at 10 μmol·L~(-1) can inhibit proliferation and promote apoptosis of THP-1 cells. The typical manifestations of apoptosis, such as cell shrinkage, nuclear rupture, and chromatin agglomerate were displayed by acridine orange staining. The decreased mRNA levels of NCOA2 and PARP1 and increased Bax/Bcl-2 ratio and the activity of pro-apoptotic protein caspase-3 were observed. Z-VAD-FMK can attenuate the apoptosis-inducing effect of miltirone. This study indicates that miltirone can inhibit the proliferation and promote the apoptosis of THP-1 cells, by down-regulating NCOA2 and PARP1, raising Bax/Bcl-2 ratio, and activating caspase-3.

Long-acting nanoparticulate DNase-1 for effective suppression of SARS-CoV-2-mediated neutrophil activities and cytokine storm.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a new strain of coronavirus not previously identified in humans. Globally, the number of confirmed cases and mortality rates of coronavirus disease 2019 (COVID-19) have risen dramatically. Currently, there are no FDA-approved antiviral drugs and there is an urgency to develop treatment strategies that can effectively suppress SARS-CoV-2-mediated cytokine storms, acute respiratory distress syndrome (ARDS), and sepsis. As symptoms progress in patients with SARS-CoV-2 sepsis, elevated amounts of cell-free DNA (cfDNA) are produced, which in turn induce multiple organ failure in these patients. Furthermore, plasma levels of DNase-1 are markedly reduced in SARS-CoV-2 sepsis patients. In this study, we generated recombinant DNase-1-coated polydopamine-poly(ethylene glycol) nanoparticulates (named long-acting DNase-1), and hypothesized that exogenous administration of long-acting DNase-1 may suppress SARS-CoV-2-mediated neutrophil activities and the cytokine storm. Our findings suggest that exogenously administered long-acting nanoparticulate DNase-1 can effectively reduce cfDNA levels and neutrophil activities and may be used as a potential therapeutic intervention for life-threatening SARS-CoV-2-mediated illnesses.

Relationship of Platelet Activation Markers and Vitamin D to Clopidogrel Resistance / 中国康复理论与实践

Objective:To explore the relationship of platelet activation markers and vitamin D to antiplatelet drug resistance in ischemic stroke patients. Methods:From June, 2017 to June, 2018, 190 patients with ischemic stroke were tested their maximum platelet aggregation rate (MPAR) induced by adenosine diphosphate (ADP) and arachidonic acid (AA), activation of platelet CD62P and P-selectin vitamin D seven to ten days after dual antiplatelet treatment (aspirin 100 mg/d + clopidogrel 75 mg/d). According to the MPAR induced by ADP and AA, the patients were divided into resistance group and sensitive group. Results:The prevalence of aspirin resistance was 1.2%, while the prevalence of clopidogrel resistance was 24.7% (47 in the resistance group and 143 in the sensitive group). The activation of platelet CD62P (t = -5.232, P < 0.001) and the prevalence of hypertension (χ2 = 4.878, P < 0.05) were more in the resistance group than in the sensitive group, while the vitamin D concentration was less (t = 3.052, P < 0.01). There was no significant difference in P-selectin between the resistance and sensitive groups (t = -0.684, P = 0.253). Logistic regression analyses showed that hypertension (OR = 5.538, 95% CI: 1.204-25.470, P < 0.05), activation of platelet CD62P (OR = 1.082, 95% CI: 1.041-1.092, P < 0.05) and vitamin D (OR = 0.848, 95% CI: 0.755-0.953, P < 0.01) were the independent related factors for clopidogrel resistance. Conclusion:Inhibition of platelet activation and supplementation of vitamin D may help to overcome the resistance of clopidogrel.

Relationship of Platelet Activation Markers and Vitamin D to Clopidogrel Resistance / 中国康复理论与实践

Objective:To explore the relationship of platelet activation markers and vitamin D to antiplatelet drug resistance in ischemic stroke patients. Methods:From June, 2017 to June, 2018, 190 patients with ischemic stroke were tested their maximum platelet aggregation rate (MPAR) induced by adenosine diphosphate (ADP) and arachidonic acid (AA), activation of platelet CD62P and P-selectin vitamin D seven to ten days after dual antiplatelet treatment (aspirin 100 mg/d + clopidogrel 75 mg/d). According to the MPAR induced by ADP and AA, the patients were divided into resistance group and sensitive group. Results:The prevalence of aspirin resistance was 1.2%, while the prevalence of clopidogrel resistance was 24.7% (47 in the resistance group and 143 in the sensitive group). The activation of platelet CD62P (t = -5.232, P < 0.001) and the prevalence of hypertension (χ2 = 4.878, P < 0.05) were more in the resistance group than in the sensitive group, while the vitamin D concentration was less (t = 3.052, P < 0.01). There was no significant difference in P-selectin between the resistance and sensitive groups (t = -0.684, P = 0.253). Logistic regression analyses showed that hypertension (OR = 5.538, 95% CI: 1.204-25.470, P < 0.05), activation of platelet CD62P (OR = 1.082, 95% CI: 1.041-1.092, P < 0.05) and vitamin D (OR = 0.848, 95% CI: 0.755-0.953, P < 0.01) were the independent related factors for clopidogrel resistance. Conclusion:Inhibition of platelet activation and supplementation of vitamin D may help to overcome the resistance of clopidogrel.

US Fine-Needle Aspiration Biopsy for Thyroid Malignancy: Diagnostic Performance of Seven Society Guidelines Applied to 2000 Thyroid Nodules.

Purpose To compare the diagnostic performance of ultrasonography (US)-based fine-needle aspiration biopsy (FNAB) criteria from seven international societies in the detection of thyroid malignancy. Materials and Methods This study included a total of 2000 consecutive thyroid nodules (≥1 cm) in 1802 patients with final diagnoses from January 2010 to May 2011. US features of the thyroid nodules were retrospectively reviewed and were classified according to the categories defined by the seven international society guidelines. The diagnostic performance of US-based FNAB criteria in the detection of thyroid malignancy and unnecessary FNAB rates were calculated and compared by using a generalized estimating equation method. Results Of the 2000 thyroid nodules, 1546 (78.3%) were benign and 454 (22.7%) were malignant, with papillary carcinoma comprising 85.5% of all malignancies. The Korean Thyroid Association/Korean Society of Thyroid Radiology (KTA/KSThR) (94.5%), National Comprehensive Cancer Network (NCCN) (92.5%), and American Thyroid Association (ATA) (89.6%) guidelines were more sensitive than those of the American Association of Clinical Endocrinologists/American College of Endocrinology/Associazione Medici Endocrinologi (AACE/ACE/AME) (80.4%), American College of Radiology (ACR) (74.7%), French Society of Endocrinology (FSE) (72.7%), and Society of Radiology in Ultrasound (SRU) (70.9%) (P < .001), while the latter guidelines had higher specificity (P < .001). The rate of unnecessary FNAB was lowest with the ACR guidelines (25.3%), followed by the FSE (29.1%), AACE/ACE/AME (32.5%), SRU (45.2%), ATA (51.7%), NCCN (54.0%), and KTA/KSThR (56.9%) guidelines. Conclusion Because the diagnostic performance of US-based FNAB criteria varies according to the individual international society guidelines, clinicians should be aware of the strengths and weaknesses of US-based FNAB criteria in the management of thyroid nodules. © RSNA, 2018 Online supplemental material is available for this article.

Evaluation of metabolism-mediated herb-drug interactions.

As the use of herbal medicines increases, the public health consequences of drug-herb interactions are becoming more significant. Herbal medicines share the same drug metabolizing enzymes and drug transporters, including cytochrome P450 enzymes (CYPs), glucuronosyltransferases (UGTs), and P-glycoprotein, with several clinically important drugs. Interactions of several commonly used herbal medicines, such as Ginko biloba, milk thistle, and St. John's wort, with therapeutic drugs including warfarin, midazolam, alprazolam, indinavir, saquinavir, digoxin, nifedipine, cyclosporine, tacrolimus, irinotecan, and imatinib in humans have been reported. Many of these drugs have very narrow therapeutic indices. As the herb-drug interactions can significantly alter pharmacokinetic and pharmacodynamic properties of administered drugs, the drugs interacting with herbal medicines should be identified by appropriate in vitro and in vivo methods. A good understanding of the mechanisms of herb-drug interactions is also essential for assessing and minimizing clinical risks. In vitro methods are useful for providing mechanistic information and evaluating multiple components in herbal medicines. This review describes major factors affecting the metabolism of herbal medicines, mechanisms of herb-drug interactions mediated by CYPs and UGTs, and several in vitro methods to assess the herb-drug interactions. Finally, drug interactions of Ginkgo biloba and St. John's wort, as representative herbal medicines, are described.

Quantitative determination of major phlorotannins in Ecklonia stolonifera.

Ecklonia stolonifera is a rich source of phlorotannins, which are responsible for the potent pharmacological effects associated with this seaweed. The purpose of this study was to develop a reversed-phase high-performance liquid chromatography method for the simultaneous determination of three major phlorotannins, eckol, dieckol, and phlorofucofuroeckol-A, in the extracts of Ecklonia stolonifera. The optimal chromatographic conditions were achieved on a Thermo Hypersil Gold C-18 column (250 x 4.6 mm i.d., 5 microm) using linear gradient elution of acetonitrile and water containing 0.1% formic acid at UV 254 nm. The separated phlorotannins were identified by liquid chromatography-mass spectrometry. The high-performance liquid chromatography method showed good linearity (r2 > 0.998), precision (1.4-9.5%), and accuracy (93.9-108.7%). The limits of detection ranged from 0.06 to 0.30 microg/mL and the lower limits of quantitation ranged from 0.2 to 1.0 microg/mL. Among phlorotannins, dieckol was the most abundant in both ethanol and ethyl acetate extracts of Ecklonia stolonifera.

Inguinal hernia repair with tension-free hernioplasty under local anesthesia.

OBJECTIVE: To evaluate the use of local anesthesia in tension-free hernioplasty in a local hospital. METHODS: The study took place at Beijing Chaoyang Hospital Affiliated to Capital Medical University, Beijing, China during the period from January 2007 to May 2008. All 110 patients who had undergone inguinal hernia repair with mesh under local anesthesia were included in the study. To increase the homogeneity of the sample, we excluded umbilical hernia repairs, parastomal hernia repairs, non-elective procedures, procedures not involving mesh, and repairs performed concurrently with another surgical procedure. We performed a retrospective review of all 110 patients' data. RESULTS: The average operating time was 45 minutes (30-70 minutes), and the average hospital stay was 3-4 days. There was no postoperative mortality in this study. No surgical site infection occurred. Two patients (18%) that suffered from a moderate scrotal hematoma had recovered after extract injection therapy was applied. The duration of incisional pain was 2-3 days, and no patient required post-operative analgesia. During the follow-up, no recurrence occurred. CONCLUSION: The use of local anesthesia in inguinal hernia repair with tension-free hernioplasty is a safe and effective alternative for inpatient treatment.

Simultaneous determination of quercetin and its glycosides from the leaves of Nelumbo nucifera by reversed-phase high-performance liquid chromatography.

The purpose of this study was to develop reversed-phase HPLC method for the simultaneous determination of the flavonoids from the leaves of Nelumbo nucifera, which have been known to exhibit antioxidant, anti-HIV, antihyperlipidemic and antiobesity effects. HPLC separation was achieved on C(18) column using gradient elution with mobile phase consisting of acetonitrile-water containing 0.1% formic acid. The separated peaks were identified by electrospray ionization mass spectrometry. The HPLC method was validated and applied for the simultaneous determination of the bioactive flavonoids from the leaves of Nelumbo nucifera. In the methanol extract, six flavonoids, quercetin, rutin, quercetin 3-O-beta-D-galactopyranoside (Qc-3-Gal), quercetin 3-O-beta-D-glucopyranoside (Qc-3-Glc), quercetin 3-O-beta-D-glucuronide (Qc-3-Gln) and quercetin 3-O-alpha-arabinopyranosyl-(1-->2)-beta-galactopyranoside (Qc-3-AraGal), were identified. Among them, Qc-3-Glc and Qc-3-Gln were found to be major component in the methanol extract of Nelumbo nucifera leaves.

Extracts of Magnoliae flos inhibit inducible nitric oxide synthase via ERK in human respiratory epithelial cells.

Nitric oxide (NO) is a marker of pulmonary inflammation. In asthma, the levels of exhaled NO are elevated and the source of this increased NO is inducible nitric oxide synthase (iNOS) within airway epithelial cells. Epimagnolin and fargesin are compounds isolated from the ethanol extract of Magnoliae flos, the seed of the Magnolia plant and are used to treat nasal congestion, headache and sinusitis in Asian countries. This study investigated whether epimagnolin and fargesin inhibit extracellular signal-regulated kinase (ERK) activation and decrease iNOS expression and NO production in stimulated human respiratory epithelial cells. An immortal Type II alveolar cell line of human origin (A549) was stimulated by cytomix (CM), composed of IL-1beta, TNF-alpha and IFN-gamma, with or without concurrent exposure to M. flos extract (epimagnolin or fargesin). CM-induced levels of NO production, iNOS expression and ERK activation were evaluated. A549 cells stimulated with CM showed increases in iNOS mRNA and protein expression, and NO synthesis. However, treatment with epimagnolin or fargesin decreased levels of iNOS mRNA and protein expression, and NO synthesis. CM stimulated a rapid increase in the activity of ERK, whereas epimagnolin and fargesin inhibited ERK phosphorylation. Epimagnolin and fargesin inhibit iNOS expression and decrease production of NO via ERK pathway in cytokine-stimulated human respiratory epithelial cells.

Rapid pharmacokinetics screening of drug candidates in vitro and in vivo / 药学学报

The paper is to report the pharmacokinetic character of a series of chemical compounds in vitro and in vivo. Metabolism stability of a series of chemical compounds was screened by using rat liver microsomes. The samples of different chemical compounds were combined and then simultaneously detected by LC-MS/MS. Compounds y13, y12 and y11 were screened out by microstability assay in vitro. The pharmacokinetics of compounds y11, y12 and y13 was evaluated by using SD rat. The plasma samples were pooled at the same time. The plasma concentrations were determined by LC-MS/MS. The pharmacokinetic character of two compounds y13, y11 was good by screening in vivo, so they were developed for further research. High-throughput screening of drug candidates in vitro and in vivo was effective, to provide information for the chemical structure information and lower the drug development risk.

PEGylation of octreotide: II. Effect of N-terminal mono-PEGylation on biological activity and pharmacokinetics.

PURPOSE: : To determine the optimal polyethylene glycol (PEG)-conjugate of octreotide by evaluating the effects of PEGylation chemistry on the biological activity and pharmacokinetic properties. METHODS: : Octreotide was chemically modified by reaction with succinimidyl propionate monomethoxy PEG (SPA-mPEG, molecular weight 2000) or succinimidyl butyraldehyde-mPEG (ALD-mPEG, molecular weight 2000 and 5000). The structural conformation of PEG-octreotides was evaluated by circular dichroism (CD), the biological activity was assessed by measuring the decrease of serum insulin-like growth factor-I levels in rats, and a pharmacokinetic study was performed after subcutaneous administration in rats. The stability against acylation was investigated with poly(D,L -lactide-co-glycolide) (PLGA). RESULTS: : ALD-mPEG was site-specific in PEGylating octreotide at the N-terminus. The mono-PEG-octreotides prepared with ALD-mPEG (mono-ALDPEG-octreotide), which alkyl bond preserves the amine's positive charge, showed complete preservation of biological activity, whereas the PEG-octreotides prepared with SPA-mPEG showed lower activity. In the CD analysis, the spectra of the mono-ALDPEG-octreotides were nearly superimposable with that of native octreotide. The mono-ALDPEG-5K-octreotide showed significantly improved pharmacokinetic properties compared with mono-ALDPEG-2K-octreotide as well as native octreotide. Both mono-ALDPEG-2K- and mono-ALDPEG-5K-octreotides were stable against acylation by degrading PLGA. CONCLUSIONS: : The mono-PEGylation of octreotide at N-terminus with ALD-mPEG produced a conjugate that is biologically and structurally active and stable against acylation by PLGA, and therefore it may serve as a candidate for somatostatin microsphere formulations.

Studies on the chemical constituents of the essential oil of Artemisia dracunculus / 中国中药杂志

<p><b>OBJECTIVE</b>To analyse the chemical constituents of Artemisia dracunculus.</p><p><b>METHOD</b>The essential oil from A. dracunculus was extracted by steam distillation and the chemical constituents were analysed by capillary GC-MS method. The relative content of each component was calculated by area normalization.</p><p><b>RESULT</b>36 peaks were separated and all of them were identified. The main chemical components were 3,7-dimethyl-1,3,7-octatriene (38.43%), 1S-alpha-pinene (36.96%), 1-methoxy-4-(2-Propenyl)-benzene (8.57%), limonene (6.33%), 1R-alpha-pinene (3.40%), etc.</p><p><b>CONCLUSION</b>The study provided solid and scientific proof for the further exploitation and utilization of A. dracunculus.</p>

Monitoring of peptide acylation inside degrading PLGA microspheres by capillary electrophoresis and MALDI-TOF mass spectrometry.

The purpose of this research was to assess the acylation reactions of peptides, salmon calcitonin (sCT), human parathyroid hormone 1-34 (hPTH1-34) and leuprolide, in poly(lactic-co-glycolic acid) (PLGA) microspheres. Capillary electrophoresis (CE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used for determining and monitoring peptide acylation and quantitating acylation products in the degrading PLGA microspheres. In the degrading PLGA microspheres of sCT and hPTH1-34, the acylation products were observed and determined to be adducts with glycolic acid units from degradable PLGA polymer by MALDI-TOF MS. In the microsphere of leuprolide, however, the acylation product was not observed even after 28 days of incubation at the release medium, which represents the different stabilities among peptides according to the primary structure. As the leuprolide contains tyrosine and serine having hydroxyl group of nucleophilic amino acids, the acylation reaction of peptide is shown to be mainly due to the primary amino groups of N-terminus or lysine residue. The complementary use of CE and MALDI-TOF MS will be useful for searching the counter measures as well as determining the peptide acylation in the manufactured formulations on the market.