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Objective:To rapidly identify the chemical constituents in Xiao Chengqitang by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap-MS）. Method:The method was established by the Waters CORTECS T3 column （2.1 mm×150 mm， 1.6 μm）， mobile phase was methanol （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-5 min， 3%-21%A； 5-20 min， 21%-36%A； 20-32 min， 36%-50%A； 32-42 min， 50%-62%A； 42-50 min， 62%-85%A； 50-60 min， 85%-95%A）， the flow rate was 0.2 mL·min<sup>-1</sup>， and the column temperature was 30 ℃. UPLC-Q-Orbitrap-MS was operated in positive and negative ion modes， the scanning range was 100-1 200 with mode of Full MS/dd-MS<sup>2</sup>， and the collision energies were 20， 40 eV. The compounds were identified by comparing with reference substances and combining with literature reports and MS database information. Result:A total of 123 components were identified in Xiao Chengqitang， including 33 flavonoids， 25 anthraquinones and anthrones， 23 phenylpropanoids， 15 tannins， 10 nitrogen-containing components and 17 other components. Among them， 32 components were determined by reference substances. Conclusion:The material basis of Xiao Chengqitang is flavonoids， anthraquinones and anthrones， phenylpropanoids， which is derived from Aurantii Fructus Immaturus，<italic> </italic>Rhei Radix et Rhizoma and Magnoliae Officinalis Cortex， respectively.
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When this paper was first published the following ethical statement was omitted in error: All animal experimental protocols were approved by the Animal Ethics Committee of Guangdong Provincial Engineering Technology Institute of Traditional Chinese Medicine (Guangzhou, Guangdong, China, Approval NO: 048483). Further, all methods were performed in accordance with the relevant guidelines and regulations. NIH mice were purchased from the Guangdong Medical Laboratory Animal Center (Guangzhou, Guangdong, China, Certificate NO.44007200031795). The authors would like to apologise for any inconvenience caused.
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Objective: To explore the effect of Sirt1 on the function of endothelial progenitor cells (EPCs) in rats with chronic obstructive pulmonary disease (COPD). Methods: A rat COPD model was established via smoking and endotoxin administration for three months. The peripheral circulating EPCs were isolated by gradient centrifugation, and their functions, cell cycle distribution, apoptosis, and Sirt1 expression were examined. The function changes of EPCs in the presence or absence of Sirt1 agonist and inhibitor were estimated; meanwhile, the expressions of Sirt1, FOXO3a, NF-κB, and p53 were also evaluated. Results: The proliferation, adhesion, and migration of EPCs decreased while the apoptosis rate was increased in the COPD rats. The expression of Sirt1 protein in EPCs of the COPD group was significantly lower than that in the control group (P<0.01). The overexpression of the Sirt1 gene using a gene transfection technique or Sirt1 agonists (SRT1720) improved the proliferation, migration, and adhesion, and decreased the apoptosis of EPC. However, Sirt1 inhibitor (EX527) decreased EPC functions in the COPD group. The effect of Sirt1 expression on EPC function may be related to reduction of FOXO3a and increase of NF-κB and p53 activity. Conclusions: Increased expression of Sirt1 can improve the proliferation and migration of EPCs and reduce their apoptosis in COPD rats. This change may be related to FOXO3a, NF-κB, and p53 signaling pathways.
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Objective: To explore the effect of Sirt1 on the function of endothelial progenitor cells (EPCs) in rats with chronic obstructive pulmonary disease (COPD). Methods: A rat COPD model was established via smoking and endotoxin administration for three months. The peripheral circulating EPCs were isolated by gradient centrifugation, and their functions, cell cycle distribution, apoptosis, and Sirt1 expression were examined. The function changes of EPCs in the presence or absence of Sirt1 agonist and inhibitor were estimated; meanwhile, the expressions of Sirt1, FOXO3a, NF-κB, and p53 were also evaluated. Results: The proliferation, adhesion, and migration of EPCs decreased while the apoptosis rate was increased in the COPD rats. The expression of Sirt1 protein in EPCs of the COPD group was significantly lower than that in the control group (P<0.01). The overexpression of the Sirt1 gene using a gene transfection technique or Sirt1 agonists (SRT1720) improved the proliferation, migration, and adhesion, and decreased the apoptosis of EPC. However, Sirt1 inhibitor (EX527) decreased EPC functions in the COPD group. The effect of Sirt1 expression on EPC function may be related to reduction of FOXO3a and increase of NF-κB and p53 activity. Conclusions: Increased expression of Sirt1 can improve the proliferation and migration of EPCs and reduce their apoptosis in COPD rats. This change may be related to FOXO3a, NF-κB, and p53 signaling pathways.
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The present study was performed to establish the UPLC fingerprints of Bolbostemmatis Rhizoma and determine the contents of three saponins by quantitative analysis of multi-components by single marker(QAMS), and provide basis for quality evaluation of Bolbostemmatis Rhizoma. The analysis was carried out on an analytical column of Waters Cortecs T3(2.1 mm×100 mm,1.6 μm)with gradient elution by acetonitrile-0.1% phosphoric acid solution, at a flow rate of 0.3 mL·min~(-1). The detection wavelength was 203 nm, the column temperature was 30 ℃ and the injection volume was 1 μL. The UPLC fingerprints of Bolbostemmatis Rhizoma were established and evaluated by similarity calculation, cluster analysis and principal component analysis. The relative calibration factors of toberoside B and toberoside C were determined with toberoside A as internal reference. The content was calculated by relative calibration factors to develop a method of QAMS. Comparing the results of QAMS with those of ESM, the accuracy and feasibility of one-eva-luation and multi-evaluation can be determined. RESULTS:: showed that the fingerprints of 19 batches of Bolbostemmatis Rhizoma have four common peaks with similarities ranging from 0.754 to 1.000. Cluster analysis and principal component analysis classified 19 batches of Bolbostemmatis Rhizoma into three categories, which was consistent with the similarity evaluation results. The relative deviation between the content of tubeicosides B and C in 19 batches of Bolbostemmatis Rhizoma determined by QAMS and ESM is less than 5.0%, indicating that there was no significant difference between the two methods. Therefore, the UPLC fingerprints combined with QAMS and similarity evaluation can be effectively used to evaluate the quality of Bolbostemmatis Rhizoma.
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Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Principal Component Analysis , Quality Control , RhizomeABSTRACT
An HPLC-ELSD method for quantitation of GCANa, CA, HDCA, CDCA and DCA in artificial cow-bezoar was developed in this study. The chromatographic separation was performed using a Wondasil CC₁₈(4.6 mmχ250 mm，5 μm) column at a column temperature of 25 °C and liquid flow-rate of 1.0 mL·min⁻¹. Acetonitrile and 0.2% formic acid solution were used as mobile phase with a linear gradient and injection volume was 10 μL. An ELSD was used with a nitrogen flow-rate of 2.3 L·h⁻¹at a drift tube temperature of 110 °C. Some chemometric methods were applied in data procession and analysis for exploration of the markers of quality difference. The accurate and simple method is suitable for the quality evaluation and the quality control of artificial cow-bezoar.
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Animals , Cattle , Bezoars , Chromatography, High Pressure Liquid , Medicine, Chinese Traditional , Quality ControlABSTRACT
A method of thin-layer fingerprinting chromatogram of artificial cow-bezoar was established with the developing solvent consisting of cyclohexane, ethyl acetate, acetic acid and methanol (2∶7∶1∶2), and 10% sulfuric acid ethanol solution sprayed as colour-developing agent. After heated at 105 ℃, TLC was recorded as an image in ultraviolet light at 366 nm which was converted into grayscale. By the gray value extracted from the grayscale, the multivariate data obtained from TLC of samples could be analyzed by chemometric method. The results indicated that samples from different manufacturers could be distinguished by this method and some specific bands were found out. All in one, this simple and practical method was suitable for the evaluation of quality difference.
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<p><b>BACKGROUND</b>The accurate measurement of the femoral anteversion (FA) angle is always a topic of much debate in the orthopedic surgery and radiology research. We aimed to explore a new FA measurement method to acquire accurate results without radiation damage using piglet model.</p><p><b>METHODS</b>A total of thirty piglets were assigned to two groups based on the age. Bilateral femora were imaged with 3.0-T magnetic resonance (MR) and 64-slice computed tomography (CT) examinations on all piglets. FA was measured on MR-three-dimensional (3D) postprocessing software with a four-step method: initial validation of the femoral condylar axis, validation of the condylar plane, validation of the femoral neck axis, and line-plane angle measurement of FA. After MR and CT examinations, all piglets were sacrificed and their degree of FA was measured using their excised, dried femora. MR, CT, and dried-femur measurement results were analyzed statistically; MR and CT measurements were compared for accuracy against each other and against the gold standard dried femur measurement.</p><p><b>RESULTS</b>In both groups, the mean FA value measured by MR was lower than that measured by CT. A statistically significant difference was observed between CT- and dried-femur measurements but not between MR- and dried-femur measurements. A higher correlation (0.783 vs. 0.408) and a higher consistency (0.863 vs. 0.578) with dried-femur measurement results were seen for MR measurements than CT measurements in the 1-week age group. However, in the 8-week age group, similar correlations (0.707 vs. 0.669) and consistencies (0.864 vs. 0.821) were observed.</p><p><b>CONCLUSIONS</b>Noninvasive MR-3D-Cube reconstruction was able to accurately measure FA in piglets. Particularly in the 1-week age group with a larger proportion of cartilaginous structures, the correlation and consistency between MR- and dried-femur measurement results were higher than those between CT- and dried-femur measurements, suggesting that MR may be a new useful examination tool for FA-related diseases in children.</p>
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Animals , Bone Anteversion , Diagnosis , Femur Neck , Pathology , Imaging, Three-Dimensional , Magnetic Resonance Imaging , Methods , Swine , Tomography, X-Ray ComputedABSTRACT
PURPOSE: The aim of this study was to identify influencing factors for sleep quality among shift-working nurses based on a three-factor scoring model that included sleep efficacy, sleep quality and daily dysfunction. METHODS: A cross-sectional survey of 513 nurses in a hospital in Shanghai, China, was conducted using a self-reported questionnaire. Sleep quality was measured by the Pittsburgh Sleep Quality Index (PSQI). Data were analyzed based on the three-factor PSQI model: Factor 1, sleep efficacy; Factor 2, sleep quality; Factor 3, daily disturbances. RESULTS: After adjusting for age, marital status, and having children, multivariate logistic regression analysis showed that participants who had previous shift work experience which was at least 6 months ago, or were currently performing shift work were significantly more likely to have poor sleep quality (PSQI > 5) than those who had never done shift work (adjusted odds ratios of 3.943 and 3.975, respectively, both p < .001). Mean scores of the three individual factors increased significantly among nurses currently performing shift work compared with those who had never done shift work (Factor 1, β = 0.61, p < .001; Factor 2, β = 1.86, p < .001; Factor 3, β = 0.45, p = .002). Mean scores of Factor 2 and Factor 3 increased significantly among nurses with previous shift work experience compared with those who had never done shift work (Factor 2, β = 1.15, p = .003; Factor 3, β = 0.52, p = .005). CONCLUSIONS: Performing current shift work and performing shift work previously were significantly associated with poor sleep quality. An appropriate arrangement and intervention strategies are needed in Chinese hospitals in order to improve sleep quality among shift-working nurses.
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Adult , Female , Humans , Male , China/epidemiology , Cross-Sectional Studies , Nurses/statistics & numerical data , Risk Factors , Sleep/physiology , Sleep Deprivation/epidemiology , Work Schedule Tolerance/physiologyABSTRACT
Objective To learn the situation of deafness gene among deaf children and to provide suggestions for intervention.Methods Twenty hot spot mutations of the common deafness genes of GJB2,GJB3,MT -RNR1,SLC26A4 for 93 deafness patients were detected by MALDI -TOF -MS,and Sanger sequencing method was used to detect the whole exon of the gene for the heterozygous mutant.Results A total of 48 cases were detected with mutation among the 93 patients using MALDI -TOF -MS,and the detection rate was 51.61%.Thirty five cases were GJB2 mutation,and the detection rate was 37.63%,in which 24 cases were homozygous mutation or compound heterozygous mutations and 11 cases were heterozygous mutation.Thirteen cases were SLC26A4 mutation,and the detection rate was 13.98%,in which 6 cases were homozygous mutation or compound heterozygous mutations and 7 cases were single heterozygous mutation.Mutation in MT -RNR1 and GJB3 gene were not detected.Among the 18 mutation cases,17 cases were detected the whole exon of the gene with mutation using Sanger sequencing,and 12 cases were detected other loci heterozygous mutation (70.59%).And a total of 42 cases were found out the cause of the deafness,and the detection rate was 45.16%.Conclusion The mutation of the common deafness gene in patients with deafness in the region has a high detection rate.The whole exon of the gene with mutation was detected,which can improve the detection rate of the cause of deafness.
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Objective: To establish a UPLC fingerprint of Compound Qima Capsule (CQC), to determine the eight contents in CQC (gastrodin, 5-hydroxymethylfurfura(5-HF), calycosin-7-O-β-D-glycoside(CG), rhoifolin, naringin, formononetin-7-O-β-D-glucoside (FG), calycosin, and formononetin), and to provide the basis for the evaluation of CQC. Methods: The Agilent Eclipse C18 (100 mm × 4.6 mm, 3.5 μm) column was used with a mobile phase of methyl acetonitrile-0.05% phosphoric acid gradient elution, the flow rate was 0.3 mL/min, the column temperature was 25℃, and the detection wavelength was 265 nm. Results: The fingerprint chromatography with good resolution and reproducibility included 22 mutual peaks, and the similarity was more than 0.98. Gastrodin, 5-HF, CG, rhoifolin, naringin, FG, calycosin, and formononetin were baseline seperated with good linearity relationships between concentration and peak areas over the linear ranges, within 0.976-29.880, 10.596-52.980, 2.697-13.485, 2.262-11.309, 40.768-203.840, 5.825-29.126, 0.372-1.858, and 1.888-9.440 μg/mL (r > 0.999 9), whose average recoveries were 1.04%, 1.30%, 1.81%, 1.41%, 1.29%, 1.01%, 1.48%, and 1.29%. The contents of 10 batches of gastrodin, 5-HF, CG, rhoifolin, naringin, FG, calycosin, and formononetin were 2.883-3.491, 1.710-3.791, 0.107-0.286, 0.157-0.346, 8.853-10.726, 0.282-0.692, 0.097-0.135, and 0.041-0.063 mg/g, respectively. Conclusion: The method is rapid, simple, and accurate, and can be used for the quality control of CQC.
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The aim of this study is to investigate the protective effects of Panax notoginseng saponins (PNS) against 6-hydroxydopamine (6-OHDA)-induced apoptosis in SH-SY5Y cells and the possible underlying mechanisms. Cell viability was examined by MTT assay. The levels of lactate dehydrogenase (LDH), reactive oxygen species (ROS), malondialdehyde (MDA) and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) were measured using the respective assay kits. Apoptosis was measured by TUNEL kit, JC-1 and ROS was measured by staining with fluorescent dyes. The activation of caspase-3 was measured with the caspase-3 assay kit. The expression of nuclear protein Nrf2 and HO-1 were determined by Western blot. PNS had significant protective effects against 6-OHDA-induced apoptosis in SH-SY5Y cells in a time- and dose-dependent manner. PNS could attenuate 6-OHDA-induced suppression of SOD, GAT, GSH-Px (PPP<0.01). Moreover, PNS pretreatment increased the expression of the nuclear Nrf2 and up-regulate HO-1. The protective effects of PNS could be inhibited by HO-1 inhibitor SnPP. In conclusion, PNS has significant protective effects against 6-OHDAinduced apoptosis in SH-SY5Y cells. The possible mechanisms of PNS are due to PNS-mediated activation of Nrf2, up-regulation of HO-1 and inhibition of oxidative stress.
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Objective To learn the detection rate of deafness predisposing genes among newborns in order to provide suggestions for the prevention of hereditary hearing loss.Methods By means of MALDI -TOF,a total of 4 025 newborns were accepted for newborn hearing and deafness predisposing genetic screening.Four common deafness predisposing genes including GJB2,GJB3,12SrRNA and SLC26A4 were detected,which included 20 hot mutation sites.Results Of the 4 025 subjects,231 were detected with deafness predisposing genes and the positive rate was 5.71%.The total rate of pathogenic mutation was 1.74‰(7 /4 025),including 1 with GJB2 235delC homozygous mutation,1 with GJB2 235delC heterozygous mutation plus 299_300 del AT heterozygous mutation and 5 with 12SrRNA homozygous mutation.The positive rate of single heterozygous mutation was 5.54% (223 /4 025).Fourteen hot mutation sites were detected.GJB2 235delC was the most common one,followed by IVS7 -2A→G.There were 109 cases with GJB2 235delC and 50 cases with IVS7-2A→G.The positive rate was 2.71% and 1.24% respectively,which was 47.19% and 21.65% of the total detection rate respectively.Conclusion The detection rate of deafness predisposing genes is high among newborns.Expanding screening sites could facilitate the detection for the carrier of deafness gene.
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Objective: To identify the flavonoids in Gegen Zhiju Soft Capsule (a soft capsule prepared with Puerariae Radix and Hoveniae Dulcis Fructus seu Semen and so on) using ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Methods: The analysis was performed on Agilent Poroshell 120 SB-C18 column (100 mm ×2.1 mm, 2.7 μm) gradient elution with a mobile phase of 0.1% formic acid aqueous solution and acetonitrile. The flow rate was at 0.4 mL/min. TOFMS was applied for qualitative analysis, and the data were collected by the negative ion mode using ESI ion source. The parameters of ion source were as follows: drying gas temperature, 300 ℃; drying gas flow rate, 10 L/min; nebulizer gas pressure, 276 kPa, sheath gas temperature, 350 ℃; sheath gas flow rate, 11 L/min; capillary voltage, 3 500 V; fragmentor voltage, 175 V. Results: Twenty flavonoid compounds were identified by TOF-MS and literature data. Relative molecular weight was concentrated within 250-650.Conclusion: The rapid separation with UPLC and Q-TOF-MS determination of the precise molecular weight information could effectively analyze and identify the flavonoid compounds in Gegen Zhiju Soft Capsule. The method is accurate, rapid, and sensitive, and could provide a reliable and effective technique for the quality control.
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<p><b>OBJECTIVE</b>To analyze the differences in the immediate effect of acupuncture on cervical spondylosis of vertebral artery type among three factors: needling technique, acupoint selection and time of needle retaining so as to option the best therapeutic program.</p><p><b>METHODS</b>Thirty-two cases were observed and randomly divided into 8 groups, 4 cases in each one. The orthogonal design of three factors and two levels was adopted. There were needling technique (electroacupuncture, reinforcing and reducing method), acupoint selection [C4-C7 Jiaji (EX-B 2) and three needles of neck: Tianzhu (BL 10), Jingbailao (EX-LHN 15), Dazhu (BL 11)] and time of needle retaining (5 min, 15 min). L8 (2(7)) orthogonal design table was arranged in the trial. The changes in mean velocity (Vm) of vertebral artery (VA) and basilar artery (BA) were observed before and after acupuncture.</p><p><b>RESULTS</b>The immediate effect of VA-BA blood flow was the most significant after electroacupuncture at C4-C7 Jiaji (EX-B 2), with continuous wave for 5 min. This method and acupoint selection greatly influenced the therapeutic results (both P<0.05).</p><p><b>CONCLUSION</b>The optimized therapeutic program of the immediate effect of acupuncture for cervical spondylosis of vertebral artery type is electroacupuncture + C4-C7 Jiaji (EX-B 2) + 5 min. The importance of different factors for the immediate effect in acupuncture treatment of cervical spondylosis of vertebral artery type is: acupoint selection > needling technique > time of needle retaining.</p>
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Acupuncture Points , Acupuncture Therapy , Regional Blood Flow , Spondylosis , Therapeutics , Vertebral ArteryABSTRACT
Based on improving experimental operating skills of students and considering the features of microbiology experimental technology and methods,we developed a new examination ways of microbiology experiment — "multimedia papers of microbiology experiments" to use among 2005 and 2006 students of sophomore(entering campus in 2005 and 2006) of biotechnology and bio-science for microbiology experiment examination.Analysis of the results of experiment examination and theoretical examination for the 2005 and 2006 students showed that the disharmony and miscorrelation between experiment examination results and theoretical examination results were overcome,and students got more interested in experimental classes,while their experimental operation is more conscious and accurate.
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Producing cellulase conditions such as different temperature,inoculum size,liquid level and pH level by Trichoderma aureoviride in the shaking bottle were studied by orthogonal design method. The results showed that the main factor affecting the producing cellulase was temperature among the orthogonal conditions.The optimum conditions were as follows:cultivating solution initial pH was 6, cultivating temperature was 28℃,inoculation size was 8%,liquid level was 40 mL in 150 mL triangle bottle,rotation speed was 170 r/min.