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ObjectiveTo establish the specific chromatogram and thin layer chromatography（TLC） identification method of Kaixinsan（KXS） samples， in order to clarify the key quality attributes and provide reference for the quality evaluation of KXS. MethodHigh performance liquid chromatography（HPLC） specific chromatogram of KXS was developed with YMC Hydrosphere C18 column（4.6 mm×250 mm， 5 μm）， the mobile phase was acetonitrile（A）-0.2% formic acid aqueous solution（B） for gradient elution（0-15 min， 2%-20%A； 15-25 min， 20%-25%A； 25-30 min， 25%-30%A； 30-45 min， 30%-31%A； 45-50 min， 31%-44%A； 50-65 min， 44%-45%A； 65-73 min， 45%-75%A； 73-95 min， 75%-100%A； 95-105 min， 100%A； 105-105.1 min， 100%-2%A； 105.1-120 min， 2%A）， the detection wavelength was 320 nm. Ultra high performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry（UHPLC-LTQ-Orbitrap MS） was used to identify the chemical components of KXS with electrospray ionization（ESI）， negative ion mode and scanning range of m/z 50-2 000. TLC identification methods for Poria and Ginseng Radix et Rhizoma in KXS were established. ResultThere were 11 common peaks in the specific chromatogram of KXS， attributed to Polygalae Radix， Poria and Acori Tatarinowii Rhizoma. Taking peak 9（α-asarone） as the reference peak， the relative standard deviations of the retention times of 15 batches of KXS samples were<0.2%. A total of 34 compounds were identified by UHPLC-LTQ-Orbitrap MS， including terpenoids， phenylpropanoids， oligosaccharides and ketones. The established TLC had good separation and was rapid， reliable， simple， feasible， suitable for the identification of Poria and Ginseng Radix et Rhizoma in KXS. ConclusionThe specific chromatogram and TLC of KXS are stable and reproducible. The material basis of KXS is basically clarified by MS， which can provide a reference for the development and quality control of KXS.
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ObjectiveTo establish the specific chromatogram and thin layer chromatography（TLC） of Qingxin Lianziyin（QXLZY） benchmark samples， in order to clarify the key quality attributes and provide a reference for the quality evaluation of QXLZY. MethodHigh performance liquid chromatography（HPLC） specific chromatogram of QXLZY benchmark samples was developed by using a YMC Hydrosphere C18 column（4.6 mm×250 mm， 5 μm） with the mobile phase of acetonitrile（A）-0.2% formic acid aqueous solution（B） for gradient elution（0-10 min， 5%-20%A； 10-20 min， 20%A； 20-25 min， 20%-24%A； 25-40 min， 24%-30%A； 40-55 min， 30%-50%A； 55-65 min， 50%-100%A； 65-75 min， 100%A； 75-75.1 min， 100%-5%A； 75.1-90 min， 5%A）， and the detection wavelength was 360 nm. Ultra-high performance liquid chromatography-linear ion trap/orbitrap mass spectrometry（UHPLC-LTQ-Orbitrap MS） with electrospray ionization（ESI） was used to identify the components of QXLZY benchmark samples by accurate relative molecular weight and multilevel MS fragment ion information， the detection conditions were positive and negative ion modes and data dependency scanning mode. TLC identification methods for Ophiopogonis Radix， Lycii Cortex， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma in QXLZY were established. ResultA total of 15 characteristic peaks were identified from Glycyrrhizae Radix et Rhizoma， Plantaginis Semen and Scutellariae Radix， and the relative standard deviations of the retention times of 15 characteristic peaks in 15 batches of QXLZY benchmark samples were≤3% with peak 8（baicalin） as the reference peak. A total of 100 compounds， including flavonoids， organic acids， saponins， amino acids and others， were identified in the benchmark samples by UHPLC-LTQ-Orbitrap MS. The established TLC had good separation and was suitable for the identification of Ophiopogonis Radix， Lycii Cortex， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma in QXLZY. ConclusionThe material basis of QXLZY benchmark samples is basically determined by MS designation and source attribution. The established specific chromatogram and TLC of QXLZY are simple， stable and reproducible， which can provide a reference for the development and quality control of QXLZY.
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ObjectiveAlmond， which is bitter in taste， contains traces of toxic substances. For the sake of the safety of prescriptions containing this medicinal material， the processing method of "soaking in boiling water" was selected. Moreover， through literature research and network pharmacology， the characteristic index of this medicinal material was determined. On this basis， a method was established for the determination of amygdalin in Qingfei Paidu Granules （QFPD） and the transfer rate of it in the processing of this prescription was monitored， aiming at improving the quality control system of QFPD. MethodThe high performance liquid chromatography conditions are as follows： YMC Triart C18 column （4.6 mm × 150 mm， 5 µm）， mobile phase of methanol-water with flow of 1.0 mL·min-1， column temperature of 35 ℃， and detection wavelength of 210 nm. ResultThe linear curve fitted well and the average recovery of amygdalin was 97.74% with RSD of 4.3%. The transfer rates of amygdalin from the medicinal material to the extract， from extract to concentrate， and from concentrate to granules were investigated with this method. The result showed that the average transfer rate from the medicinal material to the granules was （60±3.91）%. The comparison of transfer rate between the processes suggesting that the extraction of the medicinal material might be the key part influencing the prescription preparation. ConclusionThe method is simple， sensitive， reproducible， stable， and accurate， and the index is reasonable. Thus， the method can be used for the quality control of QFPD and determination of transfer rate of components in the preparation of QFPD. This study further improves the quality control standard of almond in QFPD， which can serve as a reference for the clinical application of QFPD.
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BACKGROUND A previous genome-wide association study (GWAS) identified the kinesin family member 16B (KIF16B) as a candidate gene related to sheep wool production. In this work, DNA pool sequencing and SNPscanTM high-throughput genotyping methods were used to detect single-nucleotide polymor phisms (SNPs) in the sheep KIF16B gene. The correlations between the SNPs and wool length and greasy wool yield were systematically assessed. RESULTS Forty-five SNPs were identified and 37 of them were genotyped, including 10 exon mutations, 26 intron mutations, and 1 promoter region mutation. Most of the SNPs were of medium genetic diversity and at Hardy-Weinberg equilibrium (HWE). Among them, 10 SNPs were associated with greasy wool yield and 28 SNPs impact the wool length. Five specific SNPs were found to exert significant effects on the wool length in all body parts analyzed in this study. Furthermore, linkage disequilibrium (LD) analysis was conducted among SNP loci and they were found to be significantly associated with economically important traits. Two strongly linked SNP blocks were identified within these SNPs and they might exert significant impacts on the greasy wool yield and wool length. CONCLUSIONS The identified SNPs exert significant effects on wool production and could be considered as potential DNA markers for selecting the individuals with superior phenotypes
Subject(s)
Animals , Wool/growth & development , Sheep/genetics , Sheep/growth & development , Genome-Wide Association Study/methodsABSTRACT
Objective:To study the effect of Huanglian-Jiedu Decoction on the white matter lesion of rats with focal cerebral ischemia and to explore the regulative role of the active fraction of Huanglian-Jiedu Decoction on NogoA/NgR. Methods:Male SD rats were randomly divided into sham operation group, model group, total alkaloid group, total flavonoid group, and total iridoid group. Except for the sham operation group, the rats in the other groups were used to establish the middle cerebral artery occlusion rat model by the suture method. 2 hours after modeling, rats in the total alkaloid group were given intragastric administration with 44 mg/kg total alkaloids; rats in the total flavonoid group were given intragastric administration 50 mg/kg total flavonoids; rats in the total iridoid group were given intragastric administration 80 mg/kg total iridoids, the sham operation group and the model group were intragastrically given equal volume of normal saline, once a day, for 7 consecutive days. The pathological changes of rat white matter were observed by HE staining, the pathological changes of rat myelin sheath were observed by Luxol fast blue (LFB) staining, and the expression of Amyloid precursor protein (APP), NogoA, and NgR in the internal and external capsule areas of the brain was detected by immunohistochemical staining. Real-time fluorescence quantitative PCR was used to detect the expression of NogoA and NgR in the tissues surrounding the ischemic infarct.Results:Compared with the model group, the total alkaloid group, total flavonoid group, and total iridoid group had lower pathological damage scores in the internal and external capsule areas of rats ( P<0.05 or P<0.01), increased integral optical density value of LFB staining ( P<0.01), decreased expression of APP and NogoA; the expression of NgR in the internal and external capsules of rats in the total alkaloid group and the total iridoid group decreased ( P<0.05 or P<0.01), and the expression of NgR in the inner capsule of rats in the total flavonoid group decreased ( P<0.01); the expression of NogoA (1.20 ± 0.17, 1.55 ± 0.30, 1.19 ± 0.38 vs. 2.22 ± 0.58) and NgR (1.98 ± 0.55, 1.48 ± 0.31, 1.58 ± 0.27 vs. 3.36 ± 0.41) genes in the tissues around the infarct focus of rats in the total alkaloid group, total flavonoid group and total iridoid group decreased ( P<0.01). Conclusion:The present study investigates the therapeutic effects of Huanglian-Jiedu Decoction, promoting white matter repair by decreasing the overexpression of NogoA and NgR in an experimental animal model of stroke.
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OBJECTIVE@#To investigate the effect of the combination of atrial fibrillation (AF) ablation and left atrial appendage closure (LAAC) on cardiac function and the success rate of AF ablation.@*METHODS@#We retrospectively analyzed the data of 56 patients with AF undergoing a one-stop procedure for AF ablation and LAAC in our hospital between May, 2015 and May, 2019. Propensity score matching (PSM) at the ratio of 1:1 was used to select 56 control patients undergoing AF ablation at high risk of stroke, for matching with the hybrid procedure group. The perioperative complications, thromboembolic events, recurrence of atrial arrhythmia and cardiac function were compared between the groups.@*RESULTS@#The two groups of patients were comparable for age, gender, BMI, duration and type of AF, concomitant diseases, CHA2DS2-VASc and HAS-BLED scores (@*CONCLUSIONS@#The combination of AF ablation and LAAC is safe but does not improve the success rate of AF ablation. The one-stop procedure can improve cardiac function of the patients, but AF ablation alone can achieve better improvement of cardiac function.
Subject(s)
Humans , Atrial Appendage/surgery , Atrial Fibrillation/surgery , Catheter Ablation , Retrospective Studies , Treatment OutcomeABSTRACT
Objective To develop an method for detecting the characteristic chromatograms of Qihong decoction by high-performance liquid chromatography coupled with diode arry and evaporative light-scattering detectors (HPLC-DAD-ELSD). Methods The determination was carried out with Venusil MP-C18 (250 mm × 4.6 mm, 5 μm) column,using acetonitrile-0.2％ formic acid as mobile phase with gradient elution at a flow rate of 1.0 ml/min and detected at the wave length 254 nm, 290 nm, 365 nm. The drift tube temperature for ELSD was set at 70 ℃, and the nebulizing gas flow rate was 2.8 L/min. Results There were 34 chemical compositions in characteristic chromatograms of Qihong decoction. Among them, 16 peaks came from Polygoni Orientalis Fructus, 17 from Astagali Radix, respectively. A total of 18 chemical constituents were identified. Conclusions The method was simple, steady and reliable which could be applied to the quality control of Qihong decoction.
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Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.
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To obtain chemical constituent information of rat plasma after oral administration of Huang-Lian-Jie-Du Decoction (HLJDD), a LC-FT-ICR-MS method has been established, and both positive and negative ions scan modes were include in the analysis. By comparing their retention time, high resolution mass data of HLJDD extracts, blank plasma and dosed plasma, 38 constituents, including 22 prototype compounds and 16 metabolites, were detected in rat plasma after oral administration of HLJDD. In the 22 prototype compounds, 16 constituents were determined unambiguously by comparing with references. In the analysis of metabolites, phase II reactions like glucuronidation and sulfation were the major biotransformation pathways of HLJDD. M11 was observed as the only phase I metabolite in present experiment. The results will be beneficial for the further pharmacokinetics and pharmacological evaluations of HLJDD.
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To identify the active components in Bufo melanostictus Schneider and clarify the difference between fresh and dried Venenum Bufonis, a UPLC-Orbitrap MS method has been established. The separation was performed with gradient elution of acetonitrile and water (with 0.1% formic acid) as mobile phase. By comparing their retention time and high resolution mass data of Venenum Bufonis extracts, 39 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were bufadienolides. There were significant differences in the main composition between fresh and dried Venenum Bufonis. There are fewer bufadienolides in fresh toad venom.
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Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.
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In order to clarify the metabolism pathways of scandoside methyl ester, the analysis of metabolites profiling in four kinds of liver microsomes was performed by using an ultra-performance liquid chromatography/ electrospray-tandem mass spectrometry (UPLC-ESI-MS). The data obtained from the 0 h-incubation and the 2 h-incubation were compared and analyzed. After incubation, 5 metabolites of scandoside methyl ester were found in rat, Beagles, rhesus monkey and human liver microsome. The results showed that scandoside methyl ester's major metabolic pathway in the liver microsomes is hydrolysis, oxidation and reduction reactions, and there are certain kinds differences between species. The study provides a research base for further research about iridoid compounds in vivo metabolic pathways.