Research on chemical constituents from Artemisia annua Ⅰ / 中国中药杂志

Chemical constituents were isolated and purified from the water extract of Artemisia annua by column chromatography of HP-20 macroporous resin, silica gel, ODS, Sephadex LH-20, HW-40, and semi-preparative RP-HPLC. Their structures were elucidated by physicochemical properties and spectral analyses. As a result, Fifteen compounds were isolated and identified as vitexnegheteroin M(1), sibricose A5(2), securoside A(3), citrusin D(4), annphenone(5), E-melilotoside(6), esculetin(7), scopoletin-7-O-β-D-glucoside(8), eleutheroside B_1(9), chrysosplenol D(10), patuletin-3-O-β-D-glucopyranoside(11), quercetin-7-O-β-D-glucoside(12), rutin(13), apigenin 6,8-di-C-β-D-glucopyranoside(14), isoschaftoside(15), among them, compounds 1-4 were identified from Artemisia for the first time. Additionally, the isolates were evaluated for their inhibitory effects on the production of PGE_2 in LPS-simulated RAW264.7 macrophages. The results showed that compounds 1, 2, 8, and 10-15 could reduce PGE_2 levels, to a certain extent.

Effect and mechanism of ethyl pyruvate on delayed cerebral vasospasm after subarachnoid hemorrhage / 上海交通大学学报（医学版）

Objective·To observe the degree of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) at different time points, and to investigate the effects of ethyl pyruvate (EP) on CVS following SAH and its mechanism. Methods·Fifty healthy SD rats were randomly divided into five groups i.e. sham group, SAH-3 d group, SAH-5 d group, SAH-7 d group and SAH-9 d group (n=10 in each group). Sham group was built by double cisternal injection with 0.3 mL saline each time, and SAH group was built by double cisternal injection with 0.3 mL autologous blood each time. The neurological function and degree of CVS were observed. Another forty-eight healthy SD rats were randomly divided into three groups i.e. sham+saline (equal volume) group, SAH+saline (equal volume) group and SAH+EP [100 mg/(kg·d)] group (n=16 in each group). The neurological function, degree of CVS and cell apoptosis of basilar artery were observed on day 7. The expressions of p-Akt, Bax, Bcl-2 and cleaved caspase-3 were also observed on day 7. Results·CVS significantly increased on day 3, decreased on day 5, and then significantly increased to top level on day 7, and gradually decreased on day 9. Compared with sham+saline group, the neurological scores were significantly decreased in SAH+saline group on day 7. Compared with sham+saline group, CVS and cell apoptosis of basilar artery were significantly increased in SAH+saline group on day 7. Compared with sham+saline group, the expressions of Bax and cleaved caspase-3 were significantly up-regulated, while the expressions of p-Akt and Bcl-2 were significantly down-regulated in SAH+saline group on day 7. Compared with SAH+saline group, the neurological scores significantly increased in SAH+EP group on day 7. Compared with SAH+saline group, CVS and cell apoptosis of basilar artery significantly decreased in SAH+EP group on day 7. Compared with SAH+saline group, the expressions of Bax and cleaved caspase-3 were significantly down-regulated, while the expressions of p-Akt and Bcl-2 were significantly up-regulated in SAH+EP group on day 7. Conclusion·The double hemorrhage model rat has most severely CVS on day 7. EP can attenuate vasospasm after SAH, and its mechanism may be associated with inhibition of cell apoptosis. PI3K/Akt signaling pathway may be involved, which may provide a novel therapeutic target for CVS.

HTRF-based high-throughput PGE2 release prohibition model and application in discovering traditional Chinese medicine active ingredients / 中国中药杂志

Prostaglandin (PG) E2 is an active substance in pathological and physiological mechanisms, such as inflammation and pain. The in vitro high-throughput assay for screening the inhibitors of reducing PEG2 production is a useful method for finding out antiphlogistic and analgesic candidates. The assay was based on LPS-induced PGE2 production model using a homogeneous time-resolved fluorescence(HTRF) PGE2 testing kit combined with liquid handling automation and detection instruments. The critical steps, including the cell density optimization and IC50 values determination of a positive compound, were taken to verify the stability and sensibility of the assay. Low intra-plate, inter-plate and day-to-day variability were observed in this 384-well, high-throughput format assay. Totally 5 121 samples were selected from the company's traditional Chinese medicine(TCM) material base library and used to screen PGE2 inhibitors. In this model, the cell plating density was 2 000 cells for each well; the average IC₅₀ value for positive compounds was (7.3±0.1) μmol; the Z' factor for test plates was more than 0.5 and averaged at 0.7. Among the 5 121 samples, 228 components exhibited a PGE2 production prohibition rate of more than 50%, and 23 components exhibited more than 80%. This model reached the expected standards in data stability and accuracy, indicating the reliability and authenticity of the screening results. The automated screening system was introduced to make the model fast and efficient, with a average daily screening amount exceeding 14 000 data points and provide a new model for discovering new anti-inflammatory and analgesic drug and quickly screening effective constituents of TCM in the early stage.

Monitor on influence of quality standard improvement upon Guizhi Fuling capsules efficacy / 中国中药杂志

In 2012, the preparation process and quality standard for Guizhi Fuling capsule were improved. To compare the effects and differences of capsules before (2011) and after(2012-2014) the improvement, evaluation models for intrinsic dysmenorrhea, pelvic inflammation and hysteromyoma were applied in rats. Models were induced by oxytocin, liqiud bacteria mixture and estrogen loading, respectively. The capsules (12 batchs/year, 48 bathcs in all), sampled randomly in 2011-2014, the effects were assessed using the three models. In anti-dysmenorrhea models, remarked reduction of writhing frequency, ET-1 and PGF2α content in uterus could be detected, as well as extension of writhing latency. In pelvic inflammation rats, depression of TNF-α and raise of IL-2 were induced by earh batch of capsules. In hysteromyoma model, uterine weight and smooth muscle proliferation, including E2 and P level in plasma, were lowered obviously by all batchs of capsules. Secondly, Guizhi Fuling capsules produced in 2012-2014 revealed better effectiveness than the ones manufactured in 2011. Moreover, pharmacodynamics indexes of the samples made in 2011 differed significantly between groups, which could not be observed in the ones ot 2012-2014. After tne preparation process and quality standard improvement, the effectiveness and homogeneity of Guizhi Fuling capsules were enhanced.

Main progress on studies of pharmacological activities and clinical applications of Guizhi Fuling capsule / 中国中药杂志

Guizhi Fuling capsule is a traditional Chinese medicine composed of five kinds of medicinal plants, Cinnamomi Ramulus, Poria, Moutan Cortex, Persicae Semen, and Paeoniae Radix Alba. Pharmacology studies have shown that Guizhi Fuling capsule has many activities: anti-inflammatory, analgesic, anti-tumor, regulating smooth muscle, endocrine regulation and enhancing immunity. It achieved obvious effects in the treatment of uterine fibroids, pelvic inflammatory disease, dysmenorrheal, endometriosis, ovarian cysts, breast hyperplasia and other gynecological diseases. This paper reviewed the main progress on studies of pharmacological activities and clinical applications of Guizhi Fuling capsule in recent years.

Study of effective components and molecular mechanism for Guizhi Fuling formula treatment of dysmenorrhea, pelvic inflammatory disease and uterine fibroids / 中国中药杂志

In this study, the active components and potential molecular .mechanism of Guizhi Fuling formula in treatment on dysmenorrhea, pelvic inflammation, and hysteromyoma were investigated using network pharmacological methods. Sterols and pentacyclic triterpenes, with high moleculal network degree, revealed promising effects on anti-inflammatory, analgesic, anti-tumor, and immune-regulation, according to D-T network analysis. On the other hand, the targets with high degree were involved in inflammatory, coagulation, angiopoiesis, smooth muscle contraction, and cell reproduction, which showed the novel function in anti-dysmenorrhea, pelvic inflammation, and hysteromyoma. Furthermore, the formula was indicated to play a key role in smooth muscle proliferation, inhibition of new vessels, circulation improvement, reduction of hormone secretion, alleviation of smooth muscle, block of arachidonic acid metabolism, and inflammation in uterus. Thus, the main mechanism of Guizhi Fuling formula was summarized. In conclusion, Guizhi Fuling formula was proven to alleviated dysmenorrhea, pelvic inflammation, and hysteromyoma by acting on multiple targets through several bioactive compounds, regulating 21 biological pathways.

Study on anti-inflammation and immunoloregulation effect of Guizhi Fuling capsule ingredients using high content screening / 中国中药杂志

The present study sought to investigate the anti-inflammation and immunoloregulation effect of 17 Guizhi Fuling capsule ingredients. The anti-inflammatory ingredients on LPS-induced RAW264. 7 cell injury were assessed with ELISA and immunofluorescence. The release of IL-1β, TNF-α, PGE2 were detected with ELISA and the expression of COX-2 was detected with immunofluorescence. The effects of them on promoting splenic lymphocyte proliferation were assessed with MTT and Hoechst 33342 staining method. The results showed that 15 ingredients had obviously anti-inflammatory activity on LPS- induced injury and play the immunoloregulation roles. This study suggested that the 15 ingredients may be the active ingredients on pelvic infection.

Study on the extraction recovery of ginsenoside Re in plasma by different solid-phase cartridges / 中国中药杂志

<p><b>OBJECTIVE</b>To optimize the solid-phase extraction method by comparison of the extraction recovery of ginsenoside Re plasma samples.</p><p><b>METHOD</b>After extracted by different solid-phase cartridges with water, acetonitrile, and different content methanol elution, the plasma samples were analyzed on an Zorbax SB-C18 column with acetonitrile-water gradient elution. From the recovery achieved, the best solid phase cartridge was found.</p><p><b>RESULT</b>This method consists of using 40% methanol as the wash solvent, and 80% methanol for the elution. Among the three kinds of solid-phase being tested, Waters Oasis HLB cartridge was found to be the best one.</p><p><b>CONCLUSION</b>The average extraction recovery of the Waters Oasis HLB cartridges was between 103%-113%, it can be used in the analysis of ginsenoside Re in plasma samples.</p>

Study on HPLC fingerprint of the triterpene acids in Poria cocos / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a method for HPLC fingerprint determination of the triterpene acids in Poria cocos.</p><p><b>METHOD</b>RP-HPLC, linear gradient elution and LC/MS, etc. were used to optimize the fingerprint determination method, and identify the main peaks in the HPLC fingerprint.</p><p><b>RESULT</b>A preferable method for HPLC fingerprint determination of the triterpene acids in P. cocos was established, and 9 peaks in the HPLC fingerprint were identified.</p><p><b>CONCLUSION</b>A general acquaintance of the triterpene acids in P. cocos can be obtained by using the preferable HPLC fingerprint determination method, which is useful for quality evaluation of the crud drug of P. cocos.</p>