Advances in high-resolution mass spectrometric-based data-mining technologies for detecting and characterizing the components and metabolites of Chinese materia medica / 药学学报

Research on the material basis of Chinese materia medica (CMM) is the basis for modernization of CMM. High-resolution mass spectrometry (HRMS) has become a powerful tool for the qualitative analysis of the components of CMM. Some data-mining strategies based on high-resolution mass spectrometry, such as full-information tandem mass spectrometry scanning acquisition strategy, sequential windowed acquisition of all theoretical fragment ions, mass detect filter, characteristic ion filter, mass spectral tree similarity filter, etc. have greatly promoted the elucidation of the qualitative basis of CMM. In order to provide methods for the rapid discovery and structural characterization of components and metabolites of CMM, this review summarized the advances in HRMS-based data-mining technologies for detecting and characterizing the compounds and metabolites of CMM, which includes class compounds, all compounds and metabolites.

Monitoring and quantitative analysis of trace aristolochic acid I in a Qing-Fei-Pai-Du decoction using liquid chromatography-mass spectrometry / 药学学报

The dosage of asarum in Qing-Fei-Pai-Du decoction (QFPD) is twice the dosage prescribed by the Chinese Pharmacopoeia. Due to the potential toxicity of aristolochic acid I (AAI), a limited component in asarum, the possibility that its dosage also exceeds the dosage prescribed by the Chinese Pharmacopoeia had aroused wide concern. In this study, the UHPLC-Q TOF method was used to determine the presence of AAI in QFPD. A UHPLC-QQQ method was then established to determine the content of AAI in QFPD, a reflux extract of asarum and an ultrasonic extract of 70% methanol of asarum. The results showed that the amount of AAI in the three samples was approximate 1.5, 3.2 and 9.0 μg respectively with equivalent dosages of asarum (6 g). All were obviously lower than the maximum daily limit stipulated in the Chinese Pharmacopoeia (30 μg). Therefore, we concluded that the content of AAI could be effectively reduced by using a Chinese herbal compound decoction and optimization of asarum. This assay is not only convenient, rapid, sensitive and reproducible for the trace detection of AAI in Chinese herbal compound decoction, but also useful for the rational application of asarum in QFPD.

Identification of chemical constituents in Aster tataricus by UHPLC-Q-TOF-MS / 药学学报

The root of Aster tataricus L. f. (RA) has been widely used in the clinic for moistening lung, dispelling phlegm and relieving cough because of its significant therapeutic effects on respiratory diseases. In this study, a systematic data acquisition and mining strategy was established aimed at solving the complexity of the traditional Chinese medicine using ultra high performance liquid chromatography coupled with quadruple time of flight mass spectrometry (UHPLC-Q-TOF-MS). A total of 132 chemical constituents, including 43 terpenes, 31 flavonoids, 22 organic acids, 18 peptides, 9 coumarins, 3 steroids, 3 anthraquinones and 3 aldehydes were identified or tentatively characterized, among which 59 components were confirmed by comparison with the standard references. Meanwhile, the accurate mass measurements of the identified components were all with ±5 ppm error. Therefore, this work provided not only reliable data supports for the comprehensive analysis of the chemical constituents in RA, but also provided an efficient data acquisition and mining strategy to profile the chemical constituents for other traditional Chinese medicine complex system.

Simultaneous quantification and consistency analysis of 19 representative components in Longshengzhi Capsules by HPLC-QQQ/MS / 中国中药杂志

Longshengzhi capsule consisting of 12 herbs is widely used in clinically treating cerebral ischemia during recovery period.In this study,in order to investigate the consistency of different batches of Longshengzhi capsules,a high performance liquid chromatography coupled to triple quadrupole mass spectrometry method(HPLC-QQQ/MS) was developed for the determination of 19 representative components in Longshengzhi Capsules within 9 min. Methodology validation indicated this method was simple,rapid,accurate,highly sensitive and reproducible,and it could be used for the content determination of components in Longshengzhi Capsules. The consistency analysis results showed that paeoniflorin and calycosin-7-glucoside in Longshengzhi Capsules had the highest content; RSD value of total content of 19 compounds was 5. 2% and the RSD value of main compounds such as astragaloside and calycosin-7-glucoside was all less than 15%,reflecting good consistency among different batches. This study has provided a scientific method and basis for the quality control and consistency evaluation of Longshengzhi Capsules.

Hydroxysafflor yellow A repairing the metabolic disturbances of early atherosclerosis based on fatty acid profiling / 药学学报

Atherosclerosis (AS) is a complex metabolic syndrome that seriously harms human health, and its occurrence and development are directly related to the metabolic disturbances of free fatty acids (FFA). In this study, macrophage-derived foam cells were established as the model of early AS. Therefore, the metabolic disturbances of FFA in ox-LDL induced foamy macrophages were analyzed using target metabolomics. Then the effect of hydroxysafflor yellow A (HSYA) on regulating FFA was also explored. The quantitative analysis of 27 fatty acids was obtained within 20 min based on dynamic MRM mode. Thirteen metabolic biomarkers of macrophage-derived foam cells were identified using multivariate statistical analysis. It was found that upregulation of total SFA and downregulation of C12:0, C14:0, C18:1, total MUFA were the typical metabolic features in macrophage-derived foam cells. Furthermore, HSYA displayed obvious repairing effect on C12:0, C14:0 and C18:1, which were involved in de novo fatty acid biosynthesis pathway. Oleoyl-(acyl-carrier-protein) hydrolase (OLAH), as the key enzyme in de novo fatty acid biosynthesis pathway, may be a drug target of HSYA.

Optimization of critical medium components for enhancing antibacterial thiopeptide nocathiacin I production with significantly improved quality / 中国天然药物

Nocathiacin I, a glycosylated thiopeptide antibiotic, displays excellent antibacterial activities against multidrug resistant bacterial pathogens. Previously, a novel nocathiacin I formulation for intravenous administration has been successfully developed and its aqueous solubility is greatly enhanced for clinical application. The purpose of the present study was to increase the fermentation titer of nocathiacin I and reduce or eliminate analogous impurities by screening the medium ingredients using response surface methodology. After a sysmatic optimization, a water-soluble medium containing quality-controllable components was developed and validated, resulting in an increase in the production of nocathiacin I from 150 to 405.8 mg·L at 150-L scale. Meanwhile, the analogous impurities existed in reported processes were greatly reduced or eliminated. Using optimized medium for fermentation, nocathiacin I with pharmaceutically acceptable quality was easily obtained with a recovery of 67%. In conclusion, the results from the present study offer a practical and efficient fermentation process for the production of nocathiacin I as a therapeutic agent.

Bioinformatics analysis of geranylgeranyl pyrophosphate synthase coding gene and amino acid sequence in Lamiaceae / 中国中药杂志

Geranylgeranyl pyrophosphate synthase enzyme is one of the key enzymes in the synthesis pathway of diterpenoid. Nine Lamiaceae genus GGPS synthase in Genebank was analyzed in this article. GGPS synthase the nucleic acid sequences and amino acid sequences, physicochemical properties, the signal peptide, leader peptides, transmembrane topological structure, hydrophobic, hydrophilic, subcellular localization, secondary structure, function domain, tertiary structure and evolutional relationship were predicted by using bioinformatics methods.Phylogenetic tree was constructed for the geranylgeranyl pyrophosphate synthase enzyme protein family. The results showed that GGPS amino acid sequence of the physical and chemical properties were basically identical, mainly hydrophilic protein, there existed chloroplast transit peptide, and no signal peptide and membrane structure domain, which mainly located in the chloroplast, the minor part located in mitochondria. The main secondary structures of the proteins are alpha helix and random coil. All these proteins have catalytic residues, aspartate-rich region, active site lid residues, substrate-Mg2+ binding site. The results provide theoretical reference for study on both the enzymatic characteristics of GGPS and the biosynthesis pathway of diterpenoid.

Contrasted study on pharmacokinetics of Tibetan medicine Renqing Mangjue compatible with Zuota / 中国中药杂志

To provide insights into the mechanism for the attenuate-synergistic effect of Zuota to Tibetan medicine Renqing Mangjue, a contrasted study was carried out on the pharmacokinetics of brucine and strychnine in mice plasm, which are active and toxicant ingredient in the Tibetan medicine Renqing Mangjue. LC-MS/MS was used to detect simultaneously the concentrations of brucine and strychnine in mice plasm at-different time intervals after administration parallelly and randomly, and the pharmacokinetic software Kinetica 5. 0 was selected to non-compartmental analysis (NCA) for data, and statistical analysis software SPSS 19. 0 was used for significance test on the pharmacokinetic parameters. A reliable LC-MS/MS method was established for the determination of brucine and strychnine in blood plasma, which are consistent with the requirements of the preclinical pharmacokinetic study confirmed by the methodology. The linear concentration ranges of brucine and strychnine were 0.301-104.4 µg · L(-1) (r = 0.999 5) and 0.305-106 µg · L(-1) (r = 0.999 7), respectively; The intra-day and inter-day variable coefficients were both less than 10.0% with good precision; The average extraction recoveries of brucine and strychnine were 116.23% and 112.82%, and RSD were 3.2% and 2.3% separately;The average matrix effects of brucine and strychnine were 122.48% and 116.36%, and RSD were 7.7% and 4.4%, respectively. The pharmacokinetic results showed that AUCtot of brucine and strychnine in Zuota group were both increased remarkably (P < 0.05), and the Cmax of brucine in Zuota group was about 5.25-fold higher than that of brucine in non-Zuota group (P < 0.05). The Tmax of brucine and strychnine reduced to one-eighth and one-quarter respectively compared with those in Non-Zuota group. In addition, the eliminations of brucine and strychnine in vivo were accelerated after the compatibility of Zuota. A significant difference (P < 0.05) occurred at the MRT0-t, of brucine, while the MRT0-∞ and Lz of strychnine were statistically significant upon the inspection level α = 0.1. It was found that the absorption degree of brucine and strychnine in Zuota group increased in the range of the safe dose (or concentration), while their elimination rates were accelerated, which may be one of the mechanisms for attenuate-synergistic effect of Zuota to Tibetan medicine Renqing Mangjue.

The importance of start codon of nosM in nosiheptide production / 中国天然药物

The present study was designed to investigate the effects of start codon of nosM on the biosynthesis of nosiheptide. Target genes were amplified by overlap PCR. After homologous recombination to construct engineered strains, nosiheptide production was analyzed by HPLC. Three mutants with different start codon of nosM were constructed, and nosiheptide production of each mutant was analyzed and compared. Replacement of the start codon of nosM significantly decreased the production of nosiheptide. In conclusion, start codon usage could greatly affect the biosynthetic efficiency in the biosynthetic gene cluster of nosiheptide.

Effect of ligustrazine hydrochloride on coagulation reaction and inflammation reaction in single valve replacement patients with rheumatic heart disease undergoing cardiopulmonary bypass / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the protection effect of Ligustrazine Hydrochloride (LH) on coagulation reaction and inflammation reaction in single valve replacement patients with rheumatic heart disease undergoing cardiopulmonary bypass (CPB).</p><p><b>METHODS</b>Totally 40 patients undergoing single valve replacement were recruited in the study and randomly assigned to the two groups, the treatment group and the control group, 20 in each group. In treatment group LH (3 mg/kg) was intravenously infused from the jugular vein. LH (3 mg/kg) was also added in the CPB priming. In the control group LH was replaced by equal amount of normal saline. Endothelial micro-particles (EMP) count was detected before CPB, 30 min after CPB, 1 h and 24 h after CPB finished. The coagulation reaction time (R), coagulation time (K), clotting formation velocity (alpha angle), maximum amplitude (MA), coagulation index (CI), platelet (PLT), hypersensitive C reactive protein (hs-CRP), IL-6, and IL-10 were detected before CPB, 1 h and 24 h after CPB finished.</p><p><b>RESULTS</b>There was no statistical difference in aorta arresting time, period of CPB, post-operative drainage volume, plasma transfusion volume, post-operative respirator assistant time, and hospitalization time between the two groups (P >0.05). Compared with pre-CPB in the same group, the count of EMP was much higher at 30 min after CPB and 1 h after CPB finished (P < 0.01). R and K, hs-CRP, IL-6, and IL-10 increased at 1 h and 24 h after CPB finished (P <0.01,P < 0.05). The alpha angle,.MA, CI, and PLT decreased 1 h after CPB finished (P <0.01). The a angle increased, while CI and PLT decreased 24 h after CPB finished (P <0.05). Compared with the control group in the same period, the count of EMP was lower in the treatment group 30 min after CPB and 1 h after CPB finished (P <0. 05, P <0. 01). R and K values obviously decreased in treatment group 1 hour after CPB finished (P <0. 05), while a angle, MA, CI, and PLT increased (P <0. 05, P <0. 01). hs-CRP and IL-6 decreased in the treatment group 1 h and 24 h after CPB finished (P <0.05), while IL-10 increased (P <0.05). The count of PLT increased 24 h after CPB finished in the treatment group (P <0. 05).</p><p><b>CONCLUSION</b>LH had certain protection effect on the vascular endothelium undergoing CPB, and lower excessive activation of coagulation reaction and inflammation reaction in patients undergoing CPB.</p>

A study on the mechanism of traumatic brain injury affecting the speed of bone fracture healing / 中华急诊医学杂志

Objective To evaluate the possible mechanism of traumatic brain injury (TB1) affecting the speed of bone fracture healing.Method TBI combined with unilateral tibial fracture (group A) was used to build multiple injury model and simple unilateral tibial fracture (group B),and the FOS,JUN,bFGF,and VEGF protein expression in different time points between the two groups were compared,and roentgenogram was used for the evaluation of bone healing.Results The expression of FOS,JUN,bFGF,and VEGF protein of the cerebral tissue was low in the normal rats,but was slightly enhanced in group B.There was consistence of development for FOS and JUN expression in the brain tissue in group A,reaching peak at post-TBI 3 hours,and then reducing to control level after 12 hours.The bFGF and VEGF reached peak at post-TBI 12 hours and 24 hours and reduced to control level after 72 hours,respectively.In group A and group B,an increase in the FOS,JUN protein expression around the fracture site was observed at 3 hours after injury,which reached the peak at 6 hours,and reduced to the control level after 24 hours;the comparison between group A,group B and the control group at 3 hours,6 hours and 12 hours had significant difference (P

Emergence agitation after total intravenous anesthesia for maintenance with remifentanil in children with OSAS / 上海第二医科大学学报

Objective To compare the effects of total intravenous anesthesia(TITA) by using remifentanil combined with propofol and intravenous-inhaled anesthesia by using propofol combined with sevoflurane on emergence agitation(EA) in children undergoing surgery of obstruction sleep apnea syndrome(OSAS). Methods Forty children of 3 to 7 years old scheduled for elective surgery of OSAS were randomly divided into TITA group(group T,n=20) and combined anesthesia group(group C,n=20).Patients in group T were induced with remifentanil 1 ?g/kg,midazolam 0.2 mg/kg,propofol 2.5 mg/kg and rocurolnium 0.6 mg/kg and maintained with remifentanil(0.4-0.5 ?g?kg-1?min-1) and propofol(4-6 mg?kg-1?h-1) until the end of the operation.Patients in group C were induced with midazolam 0.2 mg/kg,propofol 2.5 mg/kg and rocurolnium 0.6 mg/kg and maintained with propofol 4-6 mg?kg-1?h-1 and sevoflurane inhalation(1.2-1.4 MAC) until the end of the operation.EA was evaluated by 5-point scoring scale. Results The score of EA was significantly lower in group T than that in group C(P