Objective To establish a method and study the pharmacokinetics for concentration determination of effective components in Xiakucao Xiaoliu mixture in Normal Rat Plasma By LC-MS/MS. Methods The mobile phase was methanol-water (0.1% formic acid) system under the positive ion mode of C18 chromatographic column, gradient elution was adopted, and the flow rate was 0.3 ml/min. In the negative ion mode, the mobile phase was acetonitrile-water (0.1% formic acid) system, gradient elution, with a flow rate of 0.4 ml/min. Caffeic acid, rosmarinic acid, syringic acid, rutin in positive ion mode and Atractylodes lactone II and Atractylodes lactone III in negative ion mode were respectively determined. Normal rats were intragastrically given Xiakucao Xiaoliu Mixture 7.8 ml/kg, and blood was taken from the orbit at different time points after the administration. The blood concentration was determined by the validated LC-MS/MS method and the non-standard DAS2.0 software was used. The pharmacokinetic parameters of rats after administration were calculated by the compartment model. Results The pharmacokinetic parameters belonged to non atrioventricular model. The pharmacokinetic characteristics of the four main anti-cancer active ingredients of Caffeic acid, Rosmarinic acid, Syringic acid and Atractylodes Ⅲ in rats after administration of Xiakucao Xiaoliu Mixture were significantly different from those reported in the literature after the administration of monomers. Conclusion The established method was simple, accurate and sensitive, which could be suitable for the content determination of effective components in Xiakucao Xiaoliu mixture in Normal Rat Plasma, which would be a valuable information for the study on main anticancer active substances.

BACKGROUND: Patients with atrial fibrillation (AF) and prior stroke history have a high risk of cardiovascular events despite anticoagulation therapy. It is unclear whether catheter ablation (CA) has further benefits in these patients. METHODS: AF patients with a previous history of stroke or systemic embolism (SE) from the prospective Chinese Atrial Fibrillation Registry study between August 2011 and December 2020 were included in the analysis. Patients were matched in a 1:1 ratio to CA or medical treatment (MT) based on propensity score. The primary outcome was a composite of all-cause death or ischemic stroke (IS)/SE. RESULTS: During a total of 4.1 ± 2.3 years of follow-up, the primary outcome occurred in 111 patients in the CA group (3.3 per 100 person-years) and in 229 patients in the MT group (5.7 per 100 person-years). The CA group had a lower risk of the primary outcome compared to the MT group [hazard ratio (HR) = 0.59, 95% CI: 0.47-0.74, P < 0.001]. There was a significant decreasing risk of all-cause mortality (HR = 0.43, 95% CI: 0.31-0.61, P < 0.001), IS/SE (HR = 0.73, 95% CI: 0.54-0.97, P = 0.033), cardiovascular mortality (HR = 0.32, 95% CI: 0.19-0.54, P < 0.001) and AF recurrence (HR = 0.33, 95% CI: 0.30-0.37, P < 0.001) in the CA group compared to that in the MT group. Sensitivity analysis generated consistent results when adjusting for time-dependent usage of anticoagulants. CONCLUSIONS In AF patients with a prior stroke history, CA was associated with a lower combined risk of all-cause death or IS/SE. Further clinical trials are warranted to confirm the benefits of CA in these patients.

Objective To identify the chemical constituents of Xiakucao Xiaoliu mixture by high performance liquid chromatography-high resolution time-of-flight mass spectrometry (HPLC-TOF/MS). Methods The chromatographic separation ACE (3.0mm×150 mm) column was used. The mobile phase was methanol (A) and 0.1% formic acid (B). The gradient elution was: 0-5 min, 5% A; 5-10 min, 5%-15% A; 10-30 min, 15%-45%A; 30-40 min, 45%-70%B; 40-50 min, 70%-90%B. The injection volume was 2 μl. The flow rate was 0.4 ml/min. The column temperature was 25°C. The mass spectrometry was characterized by time-of-flight mass spectrometry, using ESI ion source. The common monitoring was in positive and negative ion mode. The reference ion was m/z 121.9856, 1033.9881. The scanning range was m/z 100-1200. Results A total of 37 chemical constituents were identified in the Xiakucao Xiaoliu mixture, 8 in the positive ion mode fragment voltage of 160 V, 28 in the negative ion mode fragment voltage of 160 V, and 19 in the fragment voltage of 260 V. Both positive and negative ions had 4 responses. The negative ion mode has 16 responses under both fragment voltages. And the ingredients were medicinal. Conclusion An effective method for the identification of the chemical constituents of Prunella vulgaris L. by HPLC-TOF/MS was established, which laid a foundation for its quality control and in-depth study in vivo.

By using multivariate statistical analysis to evaluate essential quality, and provide scientific basis for their comprehensive utilization, we established an UHPLC-QTRAP-MS/MS method for the fast, precise, efficient determination of 21 kinds of amino acids and 10 kinds of nucleosides in different species of Dendrobium. The analysis was performed on a Waters XBridge Amide column(2.1 mm×100 mm,3.5 μm) with elution by mobile phase of 0.2% formic acid in water-0.2% formic acid in acetonitrile at a flow rate of 0.2 mL·min~(-1) with the column temperature at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring(MRM) mode. The comprehensive evaluation of different species of Dendrobium was carried out by PCA and TOPSIS analysis. All 21 kinds of amino acids and 10 nucleosides showed good linearity among certain concentration range(r>0.999), the RSDs of the stability, precision, and repeatability tests were less than 3.0%. The recovery rate was in the range from 93.31% to 107.5%, and RSD was in the range of 1.1%-3.7%. The comprehensive evaluation index obtained with PCA showed that D. huoshanense was significantly higher than others regarding amino acids and D. officinale has higher nucleosides than other species. The biggest C_i difference of TOPSIS was 68.7%, and comprehensive evaluation showed that D. huoshanense produced the highest comprehensive quality. The method is precise, fast and efficient and can provide reliable basis for further researches and intrinsic quality control of Dendrobium.
Amino Acids ; Chromatography, High Pressure Liquid ; Dendrobium ; Nucleosides ; Tandem Mass Spectrometry

Objective To investigate the therapeutic effect of Xiakucao Xiaoliu mixture on Lewis lung cancer mice. Methods 30 mice with C₅₇BL/6 mouse Lewis lung cancer xenograft model were randomly divided into three groups: model control group, Xiakucao Xiaoliu mixture group (M group), cisplatin group (DDP group). M group and DDP group were administered continuously for 14 days. Through the general observation of Lewis lung cancer mice, tumor size was determined, HE staining method was used to determine the histopathological changes of tumors, and the expression of CyclinD1 and P16 in tumor tissues was determined by immunohistochemistry. Results The tumor weight of the model control group was the heaviest, and the difference was statistically significant compared with other groups. (P<0.05). Survival state and quality of life of mice had been improved to some extent in M group. The results of tumor growth curve and HE staining in each group of mice showed that the growth of tumor cells had been inhibited and normal cells had been protected. The positive expression of CyclinD1 was significantly decreased in M group and DDP group (P<0.01), but the effect of M group on the improvement of P16 positive expression was not significant. Conclusion Xiakucao Xiaoliu mixture had a good effect on inhibiting lung tumor growth.

Objective:To observe the anti-inflammatory effect of Dendrobii Huoshanense Herba. Method:Totally 60 Kunming mice were randomly divided into normal group, model group, high, middle and low-dose Dendrobii Huoshanense Herba groups (DHS-H, DHS-M, DHS-L, 4, 2, 1 g·kg^-1) and dexamethasone acetate group (DXMS, 0.01 g·kg^-1). The rats were intragastrically administered for 14 days. After the last administration for 1 h, a total of 20 μL of xylene was added to both sides of the right ear center of the mice to establish the ear swelling model. All of the mice were decapitated 1 h later, and the ear swelling inhibition rate of each group were calculated. Totally 36 healthy SD rats were divided into normal model, model group, DHS-H,DHS-M,DHS-L groups (2.8, 1.4, 0.7 g·kg^-1) and DXMS acetate group. The rats were intragastrically administered for 7 days. One hour after the last intragastric administration, a foot swelling model was established through subcutaneous injection of 10%fresh egg white in the right hind limb toe of each group. The right hind paw circumference of each rat was measured at 0, 0.5, 1, 2, 3, 4, 5 h, and the paw swelling of the rats was calculated. Totally 60 SD rats were implanted subcutaneously with sterile dry cotton balls in the bilateral groin and grouped as above. All of the rats were intragastrically administered for 14 days since the next day. After the last administration, cotton balls were taken out, and the inhibition rate of granuloma was calculated. And the contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-6 (IL-6), interferon-γ (IFN-γ) in the serum of each group were detected. Result:Compared with the normal group, the ear swelling rate, the foot swelling degree were significantly increased in the model group(P<0.01). Compared with the model group, the swelling rate of mouse ears in high, middle and low-dose Dendrobii Huoshanense Herba groups reduced significantly (P<0.01). The swelling degrees of the high and middle-dose groups were significantly decreased (P<0.05, P<0.01). The dry weight of cotton buds in each group was significantly decreased (P<0.01), with no significant change in thymus and spleen index. Compared with the normal group, the serum levels of TNF-α, IL-1β, IL-2, IL-6, and IFN-γ in the model group were higher than those in the normal group (P<0.01). And compared with model group, the levels of TNF-α, IL-1β, IL-2, IL-6, and IFN-γ in Dendrobii Huoshanense Herba group and positive drug group were decreased significantly (P<0.05, P<0.01). Conclusion:Dendrobii Huoshanense Herba could effectively inhibit acute and chronic inflammatory reactions.

Books on Chinese herbal medicines have shown that Dendrobium has the effect of nourishing Yin and reinforcing Yin,usually used for constipation induced by spleen Yin deficiency in clinical application. D. huoshanense,as an independent species among many species of Dendrobium,has no experimental studies about its effects on spleen Yin deficiency-type constipation. The purpose of this experiment was to illustrate the therapeutic effect of D. huoshanense on the constipation of spleen Yin deficiency type in rats,investigate its preliminary mechanism,and compare it with the D. officinale and D. nobile contained in the Chinese Pharmacopoeia to clarify its characteristics. The spleen Yin deficiency model was replicated in 70 rats by the composite factor method,and then the model rats were randomly divided into 7 groups: model group,Liuwei Dihuang Pills group( LWDHP),D. huoshanense high( DHS-H),medium( DHS-M),low( DHS-L) dose groups,D. nobile group( DNS),and D. officinale group( DOS),and another 10 rats were used as normal group( Normal). After 7 continuous days of administration,the fecal water content and intestine propulsion rate of each group were detected. HE staining was used to observe the pathological damage of ileum and colon in each group. Immunohistochemistry and Western blot were used to detect aquaporin 3( AQP3) expressions,while the expression levels of the somatostatin( SS) and motilin( MTL) in the ileum of each group were detected by enzyme-linked immunosorbent assay. The results showed that as compared with the model group,the rats in each drug-administered group had increased number of fecal pellets,increased fecal water content,and the increased intestinal propulsion rate( P<0. 01),while the pathological damage of the ileum and colon was significantly reduced; the expression of AQP3 protein was significantly decreased( P<0. 01); the level of MTL was significantly increased and the level of SS was decreased( P<0. 01). All DHS groups showed a good dose-effect relationship,and the same dose treatment effect was equivalent to that of DOS,but it was superior to DNS. Therefore,DHS has a significant therapeutic effect on constipation of spleen Yin deficiency type,and its mechanism may be related to intestinal motility and water-liquid metabolism,with a good therapeutic effect.
Animals ; Constipation ; drug therapy ; Dendrobium ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Intestines ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Spleen ; Yin Deficiency ; drug therapy

Aim To investigate the antitumor effects of cimigenol ( KY17 ) , a novel cycloartane triterpenoid from Cimicifuga , in human colon cancer cells (HCT116).Methods MTT assay was used to deter-mine the effect of KY17 on the proliferation of mouse embryonic fibroblasts ( MEF) and human colon cancer HCT116 cell line.Flow cytometry was employed to de-tect the effect of KY17 on HCT116 cell cycle.Fluores-cence microscopy and flow cytometry were used to ana-lyze the apoptosis .Western blot was used to detect the expression of apoptotic protein (PARP).q-PCR ana-lyzed the expression of miRNA-34a.Results The IC50 of KY17 in MEF and HCT116 cells was 27.28 μmol· L-1 and 9.31μmol· L-1 , respectively.KY17 induced HCT116 cell cycle arrest in G2/M phase and the apoptotic protein PARP cleavage . In addition, KY17 up-regulated the expression of p 53 protein and miRNA-34a.Conclusions KY17 inhibits the prolifera-tion and the cell cycle is arrested in G 2/M, inducing the apoptosis of HCT116 cells. The mechanism is probably related to miRNA-34a up-regulation and p53 activation .

This study was designed to investigate triterpenoids from the roots of Rosa laevigata Michx. The silica gel column chromatography was used to separate the chemical constituents from the roots of Rosa laevigata Michx. HPLC was used to analyze its purity and chemical constitution. Spectroscopy methods were used to determine their structures. Five constituents were isolated and identified as19α-OH-3β-E-feruloyl corosolic acid (1), 23-hydroxy-tormentic acid (2), 2α, 3β, 19α, 23-tetrahydroxy-12-en-28-oleanolic acid (3), 2α, 3α, 20β-trihydroxyurs-13 (18)-en-28-oic-acid (4), 2α, 3β, 20β-trihydroxyurs-13 (18)-en-28-oic-acid (5). Compound 1 was assigned as a new compound, compounds 4, 5 were obtained from the genus Rosa for the first time.