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Benzodiazepines(BDZs)are used in clinics for anxiolysis,anticonvulsants,sedative hypnosis,and muscle relaxation.They have high consumptions worldwide because of their easy availability and potential addiction.They are often used for suicide or criminal practices such as abduction and drug-facilitated sexual assault.The pharmacological effects of using small doses of BDZs and their detections from complex biological matrices are challenging.Efficient pretreatment methods followed by accurate and sensitive detections are necessary.Herein,pretreatment methods for the extraction,enrichment,and preconcentration of BDZs as well as the strategies for their screening,identification,and quantitation developed in the past five years have been reviewed.Moreover,recent advances in various methods are summarized.Characteristics and advantages of each method are encompassed.Future directions of the pretreatment and detection methods for BDZs are also reviewed.
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OBJECTIVE To establish HPLC finger print of Leonurus japonicus granules，and to determine the contents of 4 index components such as leonurine hydrochloride ，ferulic acid ，rutin，hyperoside. METHODS The determination was performed on Inertsil TM ODS-3 column with mobile phase consisted of acetonitrile （A）-0.1％ formic acid solution （B）in the form of gradient elution；the flow rate was 1.0 mL/min，the detection wavelength was 280 nm，the column temperature was 25 ℃，and the sample size was 5 µL. Similarity Evaluation System of Chromatogram Fingerprint of TCM （2012 edition）was used for establishing the HPLC fingerprints of 10 batches of L. japonicus granules and analyzing their similarities. By comparing with HPLC fingerprints of reference substance ，the common peaks were identified. SPSS 25.0 and SIMCA 13.0 software were used for cluster analysis and principal component analysis ；the above HPLC method was used for the content determination of 4 index components in L. japonicus granules such as leonurine hydrochloride ，ferulic acid ，rutin，hyperoside. RESULTS HPLC fingerprints of 10 batches of L. japonicus granules were established ，and 16 common peaks were matched ，and 4 peaks identified were leonurine hydrochloride （peak 6），ferulic acid （peak 13），rutin（peak 14），hyperoside（peak 16）；the similarities of 10 batches of samples were all higher than 0.970. The 10 batches of samples could be divided into four categories by cluster analysis and principal component analysis；the classification results were consistent. The contents of leonurine hydrochloride ，ferulic acid ，rutin and hyperoside were 122.10-138.82 μ g/g，9.33-10.45 μ g/g，14.12-18.95 μ g/g，5.87-8.06 μ g/g，respectively. CONCLUSIONS Established HPLC fingerprint of L. japonicus granules and the method for the content determination of 4 index components are simple and easy to operate，and have high precision and good repeatability ，which provide reference for the quality evaluation of L. japonicus granules.
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Objective:To investigate the clinicopathological characteristics, diagnosis and differential diagnosis of liposarcoma of the extremities.Methods:Fourteen patients with atypical lipomatous tumor/well-differentiated liposarcoma (ALT/WDL), dedifferentiated liposarcoma (DDL) and myxoid liposarcoma (ML) in Beijing Tiantan Hospital of Capital Medical University from March 2002 to December 2019 were collected. Fluorescence in situ hybridization (FISH) and immunohistochemical staining were used to detect the MDM2 gene amplification and expressions of S-100, CDK4 and MDM2 proteins in tumor tissues. The characteristics of different types of liposarcoma were comprehensively analyzed combined with clinical data and histopathological characteristics.Results:The age of 14 patients was (55.6±16.7) years old, with 10 males and 4 females. There were 2 cases of ALT/WDL, 6 cases of DDL, and 6 cases of ML. Immunohistochemistry showed that S-100 was positive in all patients and the patterns were scattered, partial and extensive. CDK4 and MDM2 were positive in both ALT/WDL and DDL, and weakly expressed in ML (4 cases). Vimentin was positive in all patients. MDM2 gene amplification was detected in 2 cases of ALT/WDL and 6 cases of DDL.Conclusion:Liposarcoma has a high incidence and obvious morphological diversity, accurate diagnosis is critical to the prognosis and treatment of the tumor.
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OBJECTIVE： To study the anti-inflammatory and detumescent pharmacodynamic material basis of Jingyaokang capsule， and to provide reference for secondary development， the establishment of quality control method and technological upgrading of the preparations. METHODS： The constituents of Jingyaokang capsule were extracted and separated with different solvents and macroporous adsorption resin to obtain constituent A （overall enrichment part）， constituent B （chloroform extraction part）， constituent C （water-course part） and constituent D （elution part of 60％ ethanol）. Using dexamethasone acetate as positive control， the anti-inflammatory and detumescent effects of Jingyaokang capsule and different extraction parts （constituents A， B， C， D） were investigated by mice ear edema and rat paw edema tests to screen the active fraction. UPLC-Q-TOF-MS method was used to analyze active constituent， identify compounds and attribute medicinal material. RESULTS： Anti-inflammatory and detumescent effects of constituent B （chloroform extraction part）+constituent D （elution part of 60％ ethanol） were similar to those of Jingyaokang capsule in rats or mice， indicating both had synergistic anti-inflammatory and detumescent effects and were active constituents of Jingyaokang capsule. UPLC-Q-TOF-MS detection and identification showed that constituent B contained 13 compounds as strychnine， phellodendrine， periplogenin， tetraketone alcohol， 11-carbonyl-β-mastic acid， attributing to Strychnos nux-vomica， Stephania tetrandra， Periploca sepium， Lycopodium japonicum， Boswellia carterii， etc. Constituent D contained 7 compounds as adenine， hydroxysafflower yellow A， phellodendrine， neoeriocitrin， zingibroside R1， attributing to rainworm， Carthamus tinctorius， Stephania tetrandra， Davallia mariesii， Achyranthes bidentata， etc. CONCLUSIONS： Jingyaokang capsule shows the significant anti-inflammation and detumescent effects. The chloroform extraction part is synergistic with 60％ ethanol elution part， which are the active constituents of anti-inflammation and detumescence，mainly including alkaloids， flavonoids and boswellic acids.
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OBJECTIVE： To establish the method for the rapidly non-destructive quality control of Liuwei dihuang capsule. METHODS： AOTF-NIR spectrometry was adopted. Taking 80 batches of Liuwei dihuang capsule produced by a manufacturer in recent three years as samples， HPLC chromatogram was adopted to determine the contents of loganin， morroniside， paeonol， paeoniflorin and ursolic acid； the content of water was determined according to general principles stated in 2015 edition of Chinese Pharmacopeia （part Ⅰ）. Taking 70 batches of samples as correction set， the partial least square method and the cross-validation algorithm were used to establish the NIR quantitative model of 6 indexes in Liuwei dihuang capsules with the Unscrambler quantitative analysis software. Taking residual 10 batches of samples as validation set， external validation was conducted for the model. RESULTS： The correlation coefficients （R2） of internal and external validation of loganin， morroniside， paeonol， paeoniflorin， the content of water quantitative model were all greater than 0.9； the correction of standand deviation （RMSEC） were 0.372 8， 0.025 4， 0.263 3， 0.288 5， 0.186 7 and 0.037 7； the prediction of standard deviation （RMSEP） were 0.462 2， 0.077 5， 0.472 1， 0.634 9， 0.293 4 and 0.206 9； the external verification showed that mean deviations of preclicted value to actual value were 6.04％， 6.05％， 5.87％， 6.97％， 5.62％ and 4.83％， with the mean deviation less than 10％.CONCLUSIONS：The established method can achieve rapidly non-destructive analysis Liuwei dihuang capsule.
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Objective To investigate the characteristics of histogram analysis and differences of apparent diffusion coefficient ( ADC) for normal pancreas with different diffusion gradients of diffusion weighted imaging (DWI) at 3. 0T. Methods Three unidirectional diffusion gradients (X, Y or Z direction) and three orthogonal diffusion gradient DWI data of 21 volunteers with normal pancreas were retrospectively analyzed. The histogram parameters of mean ADC, percentile, skewness, peak and the like were measured based on home-made DWI post-processing software, and the differences on histogram parameters obtained from 4 different diffusion gradients were compared by using Kruskal Wallis test. Results There was no significant difference on the mean ADC, skewness, percentile and the like for normal pancreas among 3 unidirectional diffusion gradients and tri-orthogonal diffusion gradient DWIs, but there was significant difference on the mean kurtosis value ( X:3. 16 ± 1. 44, Y:4. 72 ± 2. 26, Z:4. 47 ± 2. 1, tri-orthogonal:3. 69 ± 1. 82; χ2 = -9. 84, P=0. 02). Further analyses results showed that the ADC kurtosis of normal pancreas obtained from X-directional diffusion gradient DWI was significantly smaller than that of the direction of Y or Z, and the difference was statistically significant (P<0.05). Conclusions The ADC kurtosis of normal pancreas is related to the direction of diffusion gradient in DWI, and the ADC kurtosis of normal pancreas obtained by X-direction diffusion gradient is the smallest.
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Objective@#To investigate the characteristics of histogram analysis and differences of apparent diffusion coefficient (ADC) for normal pancreas with different diffusion gradients of diffusion weighted imaging (DWI) at 3.0T.@*Methods@#Three unidirectional diffusion gradients (X, Y or Z direction) and three orthogonal diffusion gradient DWI data of 21 volunteers with normal pancreas were retrospectively analyzed. The histogram parameters of mean ADC, percentile, skewness, peak and the like were measured based on home-made DWI post-processing software, and the differences on histogram parameters obtained from 4 different diffusion gradients were compared by using Kruskal Wallis test.@*Results@#There was no significant difference on the mean ADC, skewness, percentile and the like for normal pancreas among 3 unidirectional diffusion gradients and tri-orthogonal diffusion gradient DWIs, but there was significant difference on the mean kurtosis value (X: 3.16±1.44, Y: 4.72±2.26, Z: 4.47±2.1, tri-orthogonal: 3.69±1.82; χ2 =-9.84, P=0.02). Further analyses results showed that the ADC kurtosis of normal pancreas obtained from X-directional diffusion gradient DWI was significantly smaller than that of the direction of Y or Z, and the difference was statistically significant (P<0.05).@*Conclusions@#The ADC kurtosis of normal pancreas is related to the direction of diffusion gradient in DWI, and the ADC kurtosis of normal pancreas obtained by X-direction diffusion gradient is the smallest.
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Polyporus umbellatus,a traditional Chinese precious medicine as long been used for eliminating dampness,diuresis and have effect on cancer,getting more and more popularly in China recently. And the developmental metabolic process of the medicinal fungus,P. umbellatus,has been gotten more attention. This study is for the first time to explore the three sclerotial growth stages in P. umbellatus,named " white Polyporus"( initial phase), " grey Polyporus"( developmental phase) and " black Polyporus"( mature phase),by utilizing the de novo transcriptome assembly analysis technology. Finally,we obtained 88. 12 Gb sequence containing85 235 unigenes( ≥200 bp) assembled and 100% were annotated. We identified genes differentially expressed among the three stages of the sclerotia and screened out MFSgst,ERG4/ERG24,WD40,Rho A,CYP450,PKS,GSase and CHS1,which may contribute to the production of medicinal secondary metabolites and the defense mechanism against the environmental stress and biological invasion. We did the qRT-PCR trial to verify our results,which is in line with expectations. Our results are purposed to unearth the molecular mechanism of the accumulation of active constituents in different stages of Polyporus sclerotia which can be applied in the production and protection of Polyporus effectively.
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China , Gene Expression Profiling , Genes, Fungal , Medicine, Chinese Traditional , Polyporus , Genetics , TranscriptomeABSTRACT
Objective@#To investigate the clinicopathologic characteristics and BRAF V600E mutation of brain tumors associated with epilepsy.@*Methods@#Totally 250 patients with brain tumors associated with epilepsy were included from March 2008 to August 2017 retrospectively at Sanbo Brain Hospital, Capital Medical University.The clinical manifestations, histological features and BRAF V600E mutation results were collected and analyzed.@*Results@#There were 132 males and 118 females, and the male to female ratio was 1.1∶1.0. The age of patients ranged from 2 to 67 years(mean 22 years). The tumors had obvious local space occupying effect on MRI. The temporal lobe was the most common site (44.4%, 111/250). There were 58.4% (146/250) of ganglioglioma (GG), 24.0% (60/250) of dysembryoplastic neuroepithelial tumor (DNT), 12.8% (32/250) of pleomorphic xanthoastrocytoma(PXA), 4.0% (10/250) of angiocentric glioma (AG) and 0.8% (2/250) of papillary glioneuronal tumor (PGNT). Mixed GG, PXA and DNT morphological structures were found in 9 of patients. Among 250 cases, 35 cases were accompanied by focal cortical dysplasia(FCD). BRAF V600E was seen in 43 of 74 (58.1%) GG and 13 of 28 (46.4%) PXA. The most common pathologic grade of GG, DNT, AG and PGNT was WHO I. Some of the tumor cells from GG (34 cases) showed higher proliferative activity (WHO Ⅱ/Ⅲ). Most cases of PXA were WHOⅡand high proliferative activity was seen in nine cases.@*Conclusions@#The association of low-grade glioneuronal tumors with intractable epilepsy was well-recognized. The most common low-grade glioneuronal tumors were GG.GG may occur in any part of the central nervous system, with a predilection for temporal lobe. Each type of low-grade glioneuronal tumors has its own unique histological morphology, but some may show complex features with 2 or 3 mixed components. The occurrence of BRAF V600E mutations in GG is common, and their detection may be valuable for the diagnosis and treatment in GG.
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Objective@#To investigate the clinicopathologic characteristics of intractable epilepsy.@*Methods@#Based on the classification criteria proposed by the International League Against Epilepsy (ILAE), a retrospective analysis of the pathological characteristics was done in 822 patients who underwent epilepsy surgery in Sanbo Brain Hospital, Capital Medical University, from June 2008 to December 2012.@*Results@#The mean age of epilepsy onset was 9.9 years, mean duration of epilepsy was 11.9 years. Complex partial seizures were the main presenting features. Histopathological study showed 33 cases (4.01%) with mild forms of cortical malformations, 690 cases (83.94%) with focal cortical dysplasia (FCD) and 99 cases with others (including 39 pure hippocampal sclerosis, 20 cystosclerosis, 19 Sturge-Weber syndrome, 8 tuberous sclerosis complex, 6 without significant pathological changes, 5 gyral malformations and 2 hamartoma). Among the 690 FCD cases, 106 were FCD typeⅠ, 91 were FCD typeⅡ and 493 were FCDⅢ(Ⅲa: 160, Ⅲb: 106, Ⅲc: 26 and Ⅲd: 201).@*Conclusions@#FCDⅢd is the most common histopathological subtype causing intractable epilepsy, mainly due to focal hypoxia/ischemia in the perinatal period, which results in scarring of local brain tissue; this is followed by other isolated forms of FCD (FCDⅠand FCDⅡ), and then FCD Ⅲa and FCD Ⅲb. The reason to distinguish isolated forms of FCD (types Ⅰ and Ⅱ) from FCD Ⅲ and to subclassify FCD Ⅲ is to allow better definition of cortical dyslamination. Therefore, the pathogenic factors of intractable epilepsy can be grouped in greater details, and facilitate the diagnosis and potential curative treatment of intractable epilepsy.
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OBJECTIVE:To establish HPLC fingerprint of Zhenqi fuzheng granules. METHODS:HPLC-ELSD method was ad-opted. The determination was performed on Agilent Zorbax Eclipse XDB-C18 column with mobile phase consisted of acetonitrile-wa-ter(gradient elution)at the flow rate of 1.0 mL/min with column temperature of 40 ℃. The detector evaporation temperature was 50℃,and sample size was 10μL. Using specnuezhenide as reference,HPLC chromatograms of 7 batches of Zhenqi fuzheng gran-ules were determined. The identification and similarity evaluation of common peaks were conducted by using the TCM Chromato-graphic Fingerprint Similarity Evaluation System(2004 A edition). RESULTS:There were 13 common peaks in HPLC chromato-grams of 7 batches of samples. After validated,the similarity of 3 batches of samples in HPLC chromatograms of 7 batches of sam-ples were higher than 0.9,which were in good agreement with control fingerprints. The similarity of 4 batches of samples were <0.9,which were poorly same to control fingerprint. CONCLUSIONS:Established fingerprint can provide reference for quality evalu-ation of Zhenqi fuzheng granules.
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<p><b>OBJECTIVE</b>To correlate the presence of chromosome 1p/19q deletion with the expression of R132H mutant IDH1 status in oligodendroglial tumors, and to explore molecular markers for predicting chemosensitivity of oligodendroglial tumors.</p><p><b>METHODS</b>The study included 75 oligodendroglial tumors (38 oligodendrogliomas and 37 oligoastrocytomas). Immunohistochemistry was used to detect the expression of R132H mutant IDH1 protein, and fluorescence in situ hybridization (FISH) was employed to detect 1p/19q deletion.</p><p><b>RESULTS</b>Deletion of chromosome 1p and/or 19q was detected in 37 cases (37/75, 49.3%), among which co-deletion of 1p and 19q was seen in 34 cases (closely correlated, P < 0.01). Oligodendrogliomas WHOIIhad a slightly higher deletion rate than oligodendrogliomas WHO III, although without statistical significance. Oligodendrogliomas WHO IIand WHO III had a significantly higher deletion rate of chromosome 1p/19q than oligoastrocytomas WHO II and WHO III (P < 0.05). While combined loss of 1p/19q was always detected in oligodendrogliomas when FISH was positive, isolated 1p or 19q deletion was only found in oligoastrocytomas. The expression of R132H mutant IDH1 was detected in 51 of 75 cases (68.0%), in which oligodendrogliomas had a higher positive rate than oligoastrocytomas. Statistical analysis demonstrated a significant correlation between the expression of R132H mutant IDH1 protein and the presence of combined 1p/19q deletion in oligodendrogliomas (P < 0.05).</p><p><b>CONCLUSIONS</b>A significant correlation was observed between the expression of R132H mutant protein and 1p/19q LOH.Expression of 132H mutant IDH1 protein is the potential biomarker for predicating the presence of 1p/19q deletion in oligodendrogliomas.</p>
Subject(s)
Aged , Humans , Middle Aged , Brain Neoplasms , Genetics , Metabolism , Chromosome Deletion , Chromosomes, Human, 19-20 , Genetics , Chromosomes, Human, Pair 1 , Immunohistochemistry , In Situ Hybridization, Fluorescence , Isocitrate Dehydrogenase , Genetics , Metabolism , Mutant Proteins , Metabolism , Neoplasm Proteins , Genetics , Metabolism , Oligodendroglioma , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the feasibility of using enriched bone marrow (BM) compound with fibrin glue (FG) in repairing old radial bone defect.</p><p><b>METHODS</b>Totally 36 New Zealand rabbits were equally randomized into three groups: simple FG group, BM+FG group, and enriched BM+FG group. A 1.5-cm segmental bone defect was made at the left radial in each animal. After one month, the defect was implanted with the engineered bone. Before implantation, a compound of enriched BM with FG underwent electron microscopy, long-term culture, and bacteriological culture. Four, 8, and 12 weeks after operations, the osteogenetic effect was evaluated using X-ray observation, HE staining, or Van Gieson staining, and a semi-quantitative analysis was performed.</p><p><b>RESULTS</b>Electron microscopy showed enriched BM were compatible well with FG. No bacterial contamination or oncogenicity was observed after long-term culture. X-ray showed the repair effectiveness was significantly higher in BM+FG group and enriched BM+FG group than in simple FG group. Eight and 12 weeks after surgery, the Yang scores were significantly higher in enriched BM+FG group than in BM+FG group [(9.348±0.364évs.(7.984±0.229éìF=40.167ìP=0.001; (12.664±0.388)vs. (10.584±0.836é, F=20.3647ìP=0.004]. In addition, the Yang's scores at bone defects in BM+FG group and enriched BM+FG group were higher at the 12(th) week than in the 8(th) week. (F=36.004ìP=0.001; F=155.141ìP=0.000; respectively)The bone defects were repaired at varied degrees were histologically observed in BM+FG group and enriched BM+FG group during the observations.</p><p><b>CONCLUSION</b>Implantation of BM+FG or enriched BM+FG are effective in repairing old radial bone defects, while simple FG shows not such effect.</p>