Deep chemical identification of phytoecdysteroids in Achyranthes bidentata Blume by UHPLC coupled with linear ion trap-Orbitrap mass spectrometry and targeted isolation / 中国天然药物

Achyranthes bidentata Blume is widely used as a traditional Chinese medicine with the effects of nourishing the liver and kidneys and strengthening muscles and bones. In this work, a rapid and simple strategy was developed for characterizing phytoecdysteroids by ultra-high-performance liquid chromatography coupled with liner ion trap-Orbitrap mass spectrometry using electrospray ionization in the negative mode. As a result, 47 phytoecdysteroids were unambiguously or tentatively characterized. Among them, seven known compounds were identified according to the reference standards along with molecular formula, retention time and fragmentation patterns, while others were mostly potential new compounds. Through targeted isolation, the structures of three new compounds were determined by NMR spectra, which were consistent with LC-MS characterization. The present study provides an efficient method to deeply characterize phytoecdysteroids.

Analysis of chemical constituents in classical prescription Danggui Buxue Decoction based on RP-Q-TOF-MS and HILIC-Q-TOF-MS / 中国中药杂志

The chemical constituents of classical prescription Danggui Buxue Decoction were analyzed by reversed-phase(RP) chromatography and hydrophilic interaction chromatography(HILIC) coupled with quadrupole time-of-flight mass spectrometry. RP separation of Danggui Buxue Decoction was performed on ACQUITY UPLC HSS T3(2.1 mm×100 mm, 1.8 μm), while HILIC separation was on Waters BEH Amide(2.1 mm×100 mm, 1.7 μm). Mass spectrometry(MS) data were acquired in both negative and positive ion modes. Chemical constituents of Astragali Radix and Angelicae Sinensis Radix were searched from Reaxys and thus the in-house library was established. MS data were further analyzed by MassLynx 4.1 combined with in-house library, HMDB, Reaxys, and comparison with reference substances. In conclusion, a total of 154 compounds were identified and characterized: 16 saponins, 44 flavonoids, 10 phthalides, 7 phenylpropanoids, 15 bases and the corresponding nucleosides, 30 oligosaccharides, and 32 other compounds. Among them, 65 compounds were detected by HILIC-MS/MS. This study provides experimental evidences for the material basis research, quality control, and preparation development of Danggui Buxue Decoction and a reference method for comprehensive characterization of Chinese medicine decoctions typified by classical prescriptions.

Chemical profiling of Huashi Baidu prescription, an effective anti-COVID-19 TCM formula, by UPLC-Q-TOF/MS / 中国天然药物

Huashi Baidu prescription (HSBDF), recommended in the Guideline for the Diagnosis and Treatment of Novel Coronavirus (2019-nCoV) Pneumonia (On Trials, the Seventh Edition), was clinically used to treat severe corona virus disease 2019 (COVID-19) with cough, blood-stained sputum, inhibited defecation, red tongue etc. symptoms. This study was aimed to elucidate and profile the knowledge on its chemical constituents and the potential anti-inflammatory effect in vitro. In the study, the chemical constituents in extract of HSBDF were characterized by UPLC-Q-TOF/MS in both negative and positive modes, and the pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assays (ELISA) to determine the effects of HSBDF in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The results showed that a total of 217 chemical constituents were tentativedly characterized in HSBDF. Moreover, HSBDF could alleviate the expression levels of IL-6 and TNF-α in the cell models, indicating that the antiviral effects of HSBDF might be associated with regulation of the inflammatory cytokines production in RAW264.7 cells. We hope that the results could be served as the basic data for further study of HSBDF on anti-COVID-19 effect.

Simple and robust differentiation of Ganoderma species by high performance thin-layer chromatography coupled with single quadrupole mass spectrometry QDa / 中国天然药物

In this study, a high performance thin-layer chromatography/single quadrupole mass spectrometry QDa (HPTLC-QDa) method for robust authentication of Ganoderma lucidum, a popular and valuable herbal medicine, has been developed. This method is simple and practical, which allows direct generation of characteristic mass spectra from the HPTLC plates automatically with the application of in situ solvent desorption interface. The HPTLC silica gel plates were developed with toluene-ethyl formate-formic acid (5 : 5 : 0.2, V/V) and all bands were transferred to QDa system directly in situ using 80% methanol with 0.1% formic acid as desorption solvent. The acquired HPTLC-QDa spectra showed that luminous yellow band b3, containing ganoderic acid B/G/H and ganodeneric acid B, the major active components of Ganoderma, could be found only in G. lucidum and G. lucidum (Antler-shaped), but not in G. sinense and G. applanatum. Moreover, bands b13 and b14 with m/z 475/477 and m/z 475/491/495, respectively, could be detected in G. lucidum (Antler-shaped), but not in G. lucidum, thus allowing simple and robust authentication of G. lucidum with confused species. This method is proved to be simple, practical and reproducible, which can be extended to analyze other herbal medicines.

Relationship between the protein expression of ERCC1, BRCA, β-tubulin and K-ras and the efficacy and prognosis in advanced non-small cell lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the expression of ERCC1, BRCA1, β-tubulin and K-ras and the clinical efficacy, prognosis in advanced non-small cell lung cancer treated by platinum-based chemotherapy.</p><p><b>METHODS</b>Expression of ERCC1, BRCA1, β-tubulin and K-ras proteins were detected by immunohistochemistry in 136 patients. The relation between gene protein expression and efficacy, prognosis was analyzed.</p><p><b>RESULTS</b>(1) The efficacy of chemotherapy (ORR) in the ERCC1 negative group was better than that in the ERCC1 positive group (38.6% vs.26.4%, P < 0.017). The median survival time in the ERCC1 negative group was longer than that in the ERCC1 positive group (15 months vs. 12 months, P < 0.05). There was no significant difference in PFS. (2) The clinical stage in the BRCA1 negative group was earlier than that of positive group. There was no significant difference in ORR in either the BRCA1 negative group or BRCA1 positive group. But both MST and PFS in the BRCA1 negative group was longer than that in the BRCA1 positive group (16 months vs. 9 months, P < 0.05 and 7 months vs. 7 months, P < 0.05), respectovely. (3) There were no significant differences between the β-tubulin negative group and positive group in MST, PFS and ORR. (4) The ORR in the K-ras negative group was better than that in the K-ras positive group. There was no significant difference between the K-ras negative and positive groups in MST and PFS. (5) ERCC1 protein was an independent prognostic factor determined by multivariate analysis.</p><p><b>CONCLUSIONS</b>Multi-biomarker detection may provide important predictive value for chemotherapy efficacy and prognosis in advanced NSCLC.</p>

Investigation on levels of polybrominated diphenyl ethers in retail fish and egg products in Shenzhen / 中华预防医学杂志

<p><b>OBJECTIVE</b>To reveal the levels and distribution of polybrominated diphenyl ethers (PBDEs) in fish and egg products at retail in Shenzhen, and to evaluate the local people's exposure to PBDEs from these food.</p><p><b>METHODS</b>27 fish and egg samples were collected from supermarket and farmer's market in Shenzhen during August and October in 2008. According to the guideline of USEPA1614 method, the accelerated solvent extraction (ASE) technology was used for the extraction of PBDEs from fish and egg samples. After a series of purification processes including treatments of FMS column chromatography, acidic silica gel, silica gel and Al2O3 column, the levels of eight PBDEs congeners in the samples were determined by isotope dilution high resolution gas chromatography-high resolution mass spectrometry (HRGC-HRMS) method.</p><p><b>RESULTS</b>When BDE-209 was not taken into account, the median concentrations of ΣPBDEs in fish products was 914.7 pg/g wet weight, among which the datas for fresh water fish and sea fish were 328.2 and 1108.8 pg/g wet weight, respectively, showing a statistical significant difference (P < 0.05). BDE-47 was the predominant congener in fresh water fish and sea fish by a contribution proportion of 61% and 57%, respectively. The median concentrations of ΣPBDEs in egg products were 99.8 pg/g wet weight and the predominant congeners are BDE-47 and BDE-99, with a contribution proportion above 70%. BDE-209 was not detected in fresh water fish and the median concentration in sea fish and egg products are 243.7 and 472.6 pg/g wet weight, respectively, which caused the predominant congener changed to BDE-209 in egg products when BDE-209 was take into account. The median dietary intake of PBDEs from fish and egg products among local residents in Shenzhen was estimated as 102 ng/d.</p><p><b>CONCLUSION</b>The level of ΣPBDEs in fish and egg products in Shenzhen is relatively high. The characteristics of PBDEs pollution are quite different between fish and egg products. The level of daily dietary intake of PBDEs from fish and egg products among local residents in Shenzhen is also relatively high.</p>

The levels of dioxin in 60 human breast milk samples and the exposure risk factor analysis in Shenzhen / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the levels of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzo-p-furans (PCDD-Fs) in human breast milk of the mothers who lived in non-directly persistent organic pollutants (POPs) polluted area in Shenzhen, and the correlation of exposure risk factor was analyzed.</p><p><b>METHODS</b>From July to November in 2007, 60 primiparas by vaginal delivery after parturition 3 weeks to 2 months were sampled for breast milk who aged from 20 - 34 years old and has lived in Shenzhen non-directly POPs polluted areas over 5 years. PCDD-Fs were extracted from the frozen-dried samples with accelerated solvent extraction (ASE), cleaned up by fluid management system (FMS) and quantified by high-resolution gas chromatography-high resolution mass spectrometry (HRGC-HRMS) using isotope dilution methodology. TEQ were calculated. The correlation relationship among infant's birth weight and length, participatory's dietary, age, inhabitation period, environment and the body burden of PCDD-Fs in mother was statistically analyzed by SPSS 13.0.</p><p><b>RESULTS</b>The participants aged from 20 - 34 years old (28 years on average) and lived in Shenzhen for 5 - 29 years (10 years on average). The concentration of PCDD-Fs in 60 breast milk samples were 26.957 143 - 669.583 333 pg/g fat (mean: 7.224 817 pg/g fat, median: 84.176 062 pg/g fat), and TEQ-PCDD-Fs in samples were 2.420 793 - 29.014 277 pg/g fat (mean: 8.645 992 pg/g fat, median: 7.751 804 pg/g fat). 2, 3, 4, 7, 8-PeCDF, 1, 2, 3, 7, 8-PeCDD, 2, 3, 7, 8-TCDD were the dominant contributors to the total TEQ, which were 3.691 654 pg/g fat (42.689 378%), 2.478 315 pg/g fat (28.652 356%), and 0.980 995 pg/g fat (11.343 995%) respectively. Significant positive correlations were found among age (r = 0.26, P < 0.05), inhabitation period (r = 0.49, P < 0.05), the consumption of fish (r = 0.37, P < 0.05) and the concentrations of TEQ-PCDD-Fs in the study.</p><p><b>CONCLUSION</b>The levels of dioxin chemicals in the breast milk samples in non-directly POPs polluted areas of Shenzhen are high. Significant positive correlations were found among age, inhabitation period, the consumption of fish and the concentration of PCDD-Fs.</p>

Levels of PCDD/Fs, PCBs and PBDEs compounds in human placenta tissue / 中华预防医学杂志

<p><b>OBJECTIVE</b>To establish the methods of Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans (PCDD/Fs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) compounds determination by isotope dilution HRGC/HRMS simultaneously in human placenta tissue from mothers, and assess the human exposure risk to dioxins and PBDEs in study.</p><p><b>METHODS</b>Concentrations of 17 PCDD/Fs and 12 dioxin-like PCBs as well as 7 PBDEs were measured in human placenta tissue samples by isotope dilution HRGC/HRMS. SigmaTEQ (PCDD + PCDFs + PCBs) concentration using WHO-TEF factor and PBDEs concentration was calculated respectively. Risk assessment of mother exposure to dioxins and PBDEs was evaluated.</p><p><b>RESULTS</b>Median of SigmaTEQ (PCDD + PCDFs + PCBs) concentration for six samples was 18.15 WHO-TEQ pg/g lipid, ranging from 5.14 - 67.01 WHO-TEQ pg/g lipid. Although the median of SigmaTEQ (PCDD + PCDFs + PCBs) was lower than that of human blood of EU and Japan, and close to that of Korea and Taiwan non-exposure as reported in the literatures, the highest SigmaTEQ (PCDD + PCDFs + PCBs) concentration of placenta sample exceeded the value of high dioxins exposure area subjects in Taiwan. The dominant contributor congener for WHO-TEQ were 2, 3, 4, 7, 8-PeCDF, 1, 2, 3, 7, 8-PeCDD, PCB126, totally accounted for 65 percent of SigmaWHO-TEQ. Median and average of PBDE concentration for six samples were 2.73 ng/g lipid and 7.17 ng/g lipid, respectively, ranging from 0.95 - 25.99 ng/g lipid. BDE47 was the dominant contributor congener for the total concentration, accounted for 35 percent.</p><p><b>CONCLUSION</b>The methods of PCDD/Fs, PCBs and PBDEs compounds determined by isotope dilution HRGC/HRMS simultaneously in human placenta tissue from mothers were established successfully, and the human exposure risk to PCDD/Fs, PCBs and PBDEs should be surveyed for the donor with the highest SigmaTEQ (PCDD + PCDFs + PCBs) and PBDEs concentration of placenta sample in the future.</p>

Mechanisms of improvement of left ventricle remodeling by trans-planting two kinds of autologous bone marrow stem cells in pigs / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The necrosis of a large number of myocardial cells after acute myocardial infarction (AMI) results in a decrease of cardiac function and ventricle remodeling. Stem cell transplantation could improve cardiac function after AMI, but the involving mechanisms have not been completely understood. The present study aimed to investigate the effects of transplantation of autologous bone marrow mononuclear cells (BM-MNC) and mesenchymal stem cells (MSCs) via the coronary artery on the ventricle remodeling after AMI as well as the mechanisms of the effects of transplantation of different stem cells on ventricle remodeling.</p><p><b>METHODS</b>A total of 36 male pigs were enrolled in this study, which were divided into 4 groups: control group, simple infarct model group, BM-MNC transplantation group, and MSCs transplantation group. At 90 minutes when a miniature porcine model with AMI was established, transplantation of autologous BM-MNC ((4.7 +/- 1.7) x 10(7)) and MSCs ((6.2 +/- 1.6) x 10(5)) was performed in the coronary artery via a catheter. Ultrasound, electron microscope, immunohistochemical examination and real time reverse transcriptase-polymerase chain reaction were used respectively to observe cardiac functions, counts of blood vessels of cardiac muscle, cardiac muscle nuclear factor (NF)-kappaB, myocardial cell apoptosis, and the expression of the mRNA of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in cardiac muscles. Multivariate Logistic regression was used to analyze the correlation factors of left ventricular end-diastolic diameter (EDD).</p><p><b>RESULTS</b>The number of blood vessels in the infarct zone and around its border in the BM-MNC transplantation group was more than those in the infarct model group and MSCs group (P = 0.0001) and there was less myocardial cell apoptosis in the stem cell transplantation group than that in the infarct model group (all P < 0.01). The positive rate of NF-kappaB in the stem cell transplantation group was lower than that in the infarct model group (P = 0.001). The gene expression of VEGF in the infarct border zone of the BM-MNC group was higher than that in the MSCs group (P = 0.0001). The gene expression of bFGF in the infarct border zone in the MSCs transplantation group was higher than that in the infarct model group and the BM-MNC group (P = 0.0001). Left ventricular ejection fraction was inversely proportional to the apoptotic rate of myocardial cells and cardiac muscle NF-kappaB but positively correlated with the number of blood vessels and the expression of VEGF and bFGF in the infarct zone and infarct border zone. The Multivariate Logistic regression analysis on the factors influencing the left ventricular end-diastolic diameter after stem cell transplantation showed that the expression of VEGF mRNA in the cardiac muscles in the infarct zone, the number of apoptotic myocardial cells and the expression of NF-kappaB in the infarct border zone were independent factors for predicting the inhibitory effect on the dilation of left ventricular EDD after stem cell transplantation.</p><p><b>CONCLUSIONS</b>Transplantation of autologous BM-MNC and MSCs in pigs can improve the condition of left ventricular remodeling and recover the cardiac functions after AMI. The improvement of cardiac functions is related to the increase of blood vessels, the increased expression of VEGF and bFGF, the reduction of myocardial cell apoptosis, and the decrease of NF-kappaB level in cardiac muscle tissues after stem cell transplantation.</p>

Investigation on Levels of Polychlorinated Dibenzo-p-dioxins,Polychlorinated Dibenzo-p-furan and Polychlorinated Biphenyl in Milk in a City / 环境与健康杂志

Objective To know the levels of polychlorinated dibenzo-p-dioxins(PCDDs),polychlorinated dibenzo-p-furan (PCDFs)and polychlorinated biphenyl(PCBs)in milk and milk powder.Methods From Feb.2007 to Jun.2007,the milk and milk powder samples were collected and PCDDs,PCDFs and PCBs were extracted from the samples by Soxhlet extraction,cleaned up by FMS and quantified by HRGC-HRMS,using isotope dilution methodology.Results PCDDrFs and PCBs were detected in all samples.The mean levels of PCDD/Fs and dl-PCBs TEQ in the milk powder were 0.43 pg/g lipid(median:0.34 pg/g lipid),and in the packed milk were 3.83pg/g)lipid(median:2.04 pg/g lipid).The mean levels of PCDD/Fs and dl-PCBs TEQ in all samples were 2.13 pg/g lipid(median:0.815 pg/g lipid).The levels of PCDD/Fs and dioxin-like PCBs in the detected samples were far below the limitation of EU except of two milk samples,and the levels of PCDD/Fs and dioxin-like PCBs were higher in the packed milk than those in the milk powder.Conclusion Some of the milk and milk powder in the investigated city has been polluted by polychlorinated dibenzo-p-dioxins(PCDDs),polychlorinated dibenzo-p-furan(PCDFs)and polychlorinated biphenyl(PCBs).

Levels of polychlorinated dibenzo-p-dioxins and dibenzo-furans in sea fish samples in some sea areas in China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyse the levels of polychlorinated dibenzo-p-dioxins and dibenzo-furans (PCDD/Fs) in sea fish samples in some sea area in China surveyed by using isotope dilution HRGC/HRMS and was evaluated the local people with PCDD/Fs exposure from eating fish.</p><p><b>METHODS</b>Seven different kinds of fish and twenty fishes were sampled totally. Dibenzo-p-dioxins and dibenzo-furans were extracted from fish samples by Soxhlet extraction, concentrated and purified by FMS column chromatograph and enriched by carbon column. Confirmation and quantitative analysis at ng/kg level of PCDD/Fs was performed by HRGC/HRMS using multiple ion detection mode (MID).</p><p><b>RESULTS</b>Carp-2 was the certified reference material obtained from the NRC Institute of Canada. The concentration of 9 compounds was consistent with the certified value, and precision was evaluated in this study. The relative standard deviation was less than 15 percent for three times determination. The average concentration of 20 sea fishes was 1.48 ng/kg wet weight, the range was in 0.21-8.10 ng/kg wet weight, and the average total toxicity equivalency factor (TEQ) was 0.292 ng TEQ/kg (wet weight basis) and the range was 0.030-1.291 ng TEQ/kg for these 20 fishes. The evaluation exposure from fish for local people was 0.58 pg WHO-TEQ/kg BW.day.</p><p><b>CONCLUSION</b>The levels of PCDD/Fs was different from sample to sample, and the exposure from fish be less than WHO tolerance limit standard, however, the status for PCDD/Fs pollution should not be ignored especially when having an intake of multi-food.</p>

Effects of Self-Management Education on Quality of Life in Children with Asthma / 实用儿科临床杂志

Objective To study the effects of self-management education on quality of life in children with asthma.Methods Seventy-five out-patients and in-patients with asthma were randomlydivided into 2 groups.The educational group(38 cases)received the basic knowledge education of asthma and self-management,while the control group received only the basic knowledge education of asthma.A inquiry was conducted in all patients using a Chinese PAQLQ(pediatric asthma quality of life questionnaire)before and 6 months after the treatment.The correlation and difference between 2 groups were analyzed.Results There were no significant difference before the therapy.After 6 months's treatment,there were uptrends on scorecard in some fields in such as symptom,activity,emotion and the total score in all patients(P