Study on the Mass Spectrometry Fragmentation Regularity and Rapid Identification of Macrocyclic Polyamine Alkaloids from Tripterygium wilfordii / 中国药房

OBJECTIVE：To stud y the mass spec trometry fragmentation regularity and establish rapid identification method of macrocyclic polyamine alkaloids from Tripterygium wilfordii. METHODS ：The determination was performed on Hypersil GOLDTM C18 column with 0.1％ formic acid solution （A）-methanol（B）as mobile phase （gradient elution ）at the flow rate of 0.25 mL/min. The column temperature was 40 ℃，and sample size was 2 μL. The mass spectrometric condition included ESI，positive ion mode ， breakdown voltage of 100 V，collision energy of 35 eV，and scanning range of m/z 100-2 000. UPLC-Q-TOF-MS/MS combined with PeakView 1.2 software were used to analyze mass spectrum of 4 macrocyclic polyamine alkaloids control as celafurine ， celabenzine，celacarfurin，celacinnine and total extract of T. wilfordii ，and summarized the mass fragmentation regularity of alkaloids. According to retention time ，accurate molecular weight and secondary mass fragment ion information ，referring to the above mass spectrometry fragmentation regularity ，macrocyclic polyamine alkaloids in the total extract of T. wilfordii were quickly identified，and whether it was a new compound was determined by consulting Scifinder database and literature comparison. RESULTS：There were three characteristic cleavage fragments of macrocyclic polyamine alkaloids ，i.e. m/z 160.11（base peak ），m/z 188.10 and m/z 100.07 in positive ion mode. According to the precise molecular weight and mass spectrometry fragmentation regularity，10 macrocyclic polyamine alkaloids were identified rapidly ，and compound 1-6，8 were 2-phenyl-1，5，9-triazacy- clotridecan-4-one， 9-acetyl-2-phenyl-1，5，9-triazacyclotridecan-4-one， 9-nicotinoyl-2-phenyl-1， 5， 9-triazacyclotridecan-4-one， celafurine，celabenzine，celacarfurine，celacinnine，respectively. Compounds 3，7，9 and 10 might be new compounds. CONCLUSIONS：The macrocyclic polyamine alkaloids canform three common characteristic fragment ions ，m/z 160.11（base peak ），m/z 188.10 and m/z 100.07，which can be used for the rapid identification of these active components.

Establishment of prediction model for uterine scar healing after cesarean section / 中国医师杂志

Objective:To analyze the related factors of poor uterine scar healing after cesarean section, to establish and evaluate a nomogram model for predicting the risk of poor uterine scar healing after cesarean section.Methods:A total of 170 pregnant women who underwent cesarean section in Hunan Provincical Maternal and Child Care Hospital from April 2019 to May 2020 were prospectively selected as the research objects, and they were divided into poor healing group (48 cases) and good healing group (122 cases) according to the uterine scar healing situation after cesarean section. Logistic regression model was used to analyze the related factors of poor uterine scar healing after cesarean section. The nomograph model for predicting the risk of poor uterine scar healing after cesarean section was drawn by using rms package in R language (R3.6.3). The nomogram model was evaluated and verified by receiver operating characteristic curve (ROC), calibration curve and Hosmer-Lemeshow goodness of fit test.Results:Logistic regression model showed that prenatal body mass index (BMI), the number of cesarean section, premature rupture of membranes, incision position near the cervical orifice, operation time were the independent risk factors of poor uterine scar healing after cesarean section ( P<0.05). According to the above results of logistic regression analysis, a nomogram model was drawn to predict the risk of poor uterine scar healing after cesarean section. The ROC results showed that the area under curve (AUC) of this nomogram model to predict the risk of poor uterine scar healing after cesarean section was 0.902. The calibration curve was a straight line with a slope close to 1 and Hosmer-Lemeshow goodness of fit test (χ 2=5.912, P=0.657) showed that the nomogram model has good consistency and good calibration. Conclusions:The nomogram model for predicting the risks of poor uterine scar healing after cesarean section is established based on the five independent factors of maternal prenatal BMI, number of cesarean section, premature rupture of membranes, incision position near the cervical orifice and operation time, which has good accuracy and differentiation.

Separation and Identification of Chemical Components in Ethyl Acetate Fraction and Water Fraction from Tripterygium wilfordii / 中国药房

OBJECTIVE： To separate and identify chemical components in ethyl acetate fraction and water fraction from Tripterygium wilfordii， and to provider basis for further pharmacological study. METHODS： The ethyl acetate fraction and water fraction from T. wilfordii were separated and purified by MCI GEL-CHP 20P column chromatography， C18 RP silica gel column chromatography， Sephadex LH-20 gel column chromatography and HPLC. The structures of compounds were analyzed and identified by 1H-NMR， 13C-NMR and physicochemical properties. RESULTS： Two compounds were isolated from ethyl acetate fraction of T. wilfordii， namely orthosphenic acid （compound 1）， dibutylphthalate （compound 2）. Eight glucosides were isolated from water extract of T. wilfordii， namely 2，6-dimethoxy-4-hydroxymethyl-phenyl-1-O-beta-D-glucopyranoside （compound 3）， 2，6-dimethoxy-4-hydroxyphenol-1-O-β-D-glucoside（compound 4）， 4-hydroxy-1-（2-hydroxyethyl）-phenyl-3-O-β-D-glucopyranoside （compound 5）， 3，4-dimethoxy-phenyl-1-O-β-D-glucopyranoside （compound 6），β-adenosine （compound 7）， ligustrin （compound 8）， epicatechin-8-C-β-D-galactoside （compound 9） and 2-hydroxynaringenin-7-O-β-glucoside （compound 10）. CONCLUSIONS： Chemical components of ethyl acetate fraction and water fraction are separated and identified from T. wilfordii.

Study on Thermal Analysis of Tripterygium wilfordii and Its Counterfeits / 中国药房

OBJECTIVE:To provide reference for the identification,processing,processing technology and research for ther-mal decomposition products. METHODS:Thermogravimetry- derivative thermogravimetry (TG-DTG) and differential thermal-de-rivative differential thermal (DTA-DDTA) analysis were conducted to scan thermal spectrum of tripterygium from 3 places and T. hypoglaucum,T. regelii,common counterfeits Actinidia arguta and Vitis quinquangularis,determine 7 medicinal materials,its dregs and extractum,and analyze differential spectrum of T. wilfordii and dregs. RESULTS:The TG-DTG and DTA-DDTA thermal spectrum were not only basically similar to each other but also to their dregs. There was a singlet main characteristics peak at about (299±3)℃ only in the DTG curves of extracts of tripterygium from 3 places and T. hypoglaucum extractum with similar intensity, while T. regelii,counterfeits A. arguta and V. quinquangularis showed multiple peaks,and (299 ± 3)℃ was not the main peak. CONCLUSIONS:The thermal spectrum can not distinguish tripterygium and its counterfeits;extractum DTG curves can do it;suit-able processing temperature of tripterygium was 200-380℃. The study can provide reference for the identification,processing,pro-cessing technology and research for thermal decomposition products.

Effects of luteolin on CaM-CaMPK signaling pathway in hippocampus in epileptic rats / 国际中医中药杂志

Objective To explore the effects of luteolin on cognition function in pentylenetetrazol (PTZ)-induced epileptic rats and related mechanism.Methods Fifty male SD rats were randomly divided into a normal control group(n=8), a model group(n=12), and groups of 25, 50 mg/kg luteolin(both ofn=11), as well as 100 mg/kg luteolin group(n=8). Those rats were given different doses of luteolin (25, 50 and 100 mg/kg, daily, intragastric administration) for 36 consecutive days. Similarly, rats of the normal control group and the model group were given 0.5% sodium carboxymethyl cellulose suspension liquid via intragastric administration. Thirty minutes later, a model of epilepsy was induced using PTZ (40 mg/kg, daily) via intraperitoneal injection except the control group. Learning and memory of rats were evaluated by Morris water maze and novel objective recognition trials(including escape latency and recognition index). The levels of CaM and CaMPK were determined by ELISA methods, and expression of Ras proteins in the hippocampus were detected by Western Blot.Results Compared with the model group, luteolin treatment groups significantly shorten the escape latency(28.51 ± 3.84 s, 19.77 ± 5.41 s, 14.86 ± 2.76 svs. 37.08 ± 5.18 s) in the Morris water maze, and increased recognition index(18.77% ± 2.02%, 25.06% ± 4.32%, 31.92% ± 2.65%vs. 13.87% ± 2.14%) in the novel objection trial(P<0.05 orP<0.01). Meanwhile, CaM(140.33 ± 13.52 ng/L, 124.26 ± 9.97 ng/L, 113.52 ± 11.57 ng/Lvs. 158.36 ± 10.68 ng/L) and CaMPK(8.25 ± 1.37 ng/ml, 7.69 ± 0.84 ng/ml, 6.74 ± 0.93 ng/mlvs. 9.87 ± 1.02 ng/ml) were significantly decreased(P<0.05 orP<0.01). What’s more, the expression of Ras proteins(0.99 ± 0.08, 0.76 ± 0.07, 0.52 ± 0.07vs. 1.58 ± 0.12) was obviously decreased compared with the model group(P<0.05 orP<0.01).Conclusion Luteolin could effectively improve the cognition dysfunction of epileptic rats, and the mechanism might be relevant to regulate the CaM-CaMPK signaling pathway via down-regulation of CaM, CaMPK, as well as Ras protein.

Study on the Chemical Constituents of Rabdosia rubescens / 中国药房

OBJECTIVE:To study the chemical constituents of Rabdosia rubescens. METHODS:The chemical constituents of R. rubescens were isolated and purified by silica gel,MCI GEL-CHP 20P,Sephadex LH-20 column and recrystallization,and their structure was identified on the base of spectral analysis and physicochemical properties. RESULTS:8 compounds were isolated in R. rubescens and identified as 24ζ-methyl-5α-lanosta-25-ketone(1),maoyecrystal F(2),rosthorin A(3),β-sitosterol(4),daucosterol (5),heptacosanoic acid (6),oridonin (7) and ponicidin (8). CONCLUSIONS:Compound 1 is isolated from R. genera for the first time and compound 3,6 are isolated from this plant for the first time. The research lays a certain foundation for the quality evaluation of R.rubescens.

Influences on Pharmacokinetics of Hydrochloric Berberine in Rats Administrated Gangtai with 3 Routes / 中国中医药信息杂志

Objective To observe and compare the pharmacokinetics of hydrochloric berberine in rats administrated Gangtai by 3 routes.Method Rats were taken orally or administrated Gangtai by abdomen skin or rectal,and their serum concentrations of hydrochloric berberine at different time(0～30 h)were determined with HPLC method.Results The main pharmacokinetics parameters of hydrochloric berberine by abdomen skin,rectal and oral administration:Cmax was 127.4,269.4,127.9 g/L respectively,Tmax was 1,0.5,0.5 h respectively,AUC0-30 h was 161 1.44,105 5.61,765.88 g/(L?h)respectively.Conclusion The pharmacokinetics parameters of Gangtai in rats provided academic evidence for clinical application of Gangtai.