Study on Formation Mechanism of National Drug Standards Based on Standard Investigation of Traditional Chinese Medicine Dispensing Granules / 中国实验方剂学杂志

This paper, taking the formulation of national drug standards for traditional Chinese medicine (TCM) dispensing granules as a case study, explores the improvement of the formation mechanism of national drug standards, and promotes the reform of streamline administration, delegate powers, and improve regulation and services of national standards management, so as to release the vitality of the research and development of standards of drug manufacturers. After nearly two decades of pilot production of TCM dispensing granules, a large number of researches and discussions have been conducted on the formulation of unified standards of TCM dispensing granules from manufacturing enterprises to national standard administration departments, it was found that this work was difficult on the basis of the original drug standard formation mechanism. The authors tried to improve and innovate the formation mechanism of national drug standards, to provide methods and ideas for the formulation and unification of national standards for TCM dispensing granules, and to provide references for the formulation of other national drug standards.

Protective Effect of Tongxinluo Capsule on Cerebral Ischemia-reperfusion Injury and Its Mechanism of Action / 中国实验方剂学杂志

Objective:To study the protective effect of Tongxinluo capsule on cerebral ischemia-reperfusion injury, and explore the mechanism of Tongxinluo capsule in treating cerebral ischemia through network pharmacology. Method:The C57BL/6 mouse middle cerebral artery occlusion(MCAO) model was established by improved suture method and divided into sham operation group, model group, low, medium and high-dose Tongxinluo groups (crude drug 1,2,4 g·kg-1, intragastric administration), Aspirin group (2.055 g·L-1, intraperitoneal injection). Then, neurological function score and 2,3,5-triphenyltetrazole chloride(TTC) staining method were used to determine the infarct size of mice at 24, 48, 72 h by cerebral ischemia-reperfusion. First, chemical constituents of Tongxinluo capsule were screened from the BATMAN-TCM database, and the targets were analyzed. Then, gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were performed, and traditional Chinese medicine(TCM)-active ingredient-target network was constructed. Finally, the multi-dimensional pharmacological mechanism of Tongxinluo capsule in the treatment of cerebral ischemia was predicted. Result:Longa score, HE staining and TTC staining all suggested that Tongxinluo capsule could alleviate brain injury in mice after cerebral ischemia and reperfusion, and the improvement degree of Tongxinluo capsule on brain injury was gradually enhanced with the increase of Tongxinluo capsule dose. A total of 132 active components and 240 intersection targets, including cyclic adenosine monophosphate(cAMP)-dependent protein kinase catalytic subunit alpha (PRKACA), adenylate cyclase 1(ADCY1), serine/threonine kinase 1 (Akt1), dopamine receptor D2 (DRD2) and discs large homolog 4 (DLG4) were screened from 12 TCM in Tongxinluo capsule. GO was enriched in cationic channel activity, ion gated channel activity, gate channel activity, neurotransmitter receptor activity, ion channel activity, etc. KEGG was enriched in cAMP signaling pathway, calcium signaling pathway, neuroactive ligand-receptor interaction, cyclic guanosine monophosphate(cGMP)/protein kinase G(PKG) signaling pathway and dopaminergic synaptic signaling pathway. Conclusion:Tongxinluo capsule can alleviate brain damage in mice with cerebral ischemia-reperfusion, and achieve brain protection through multiple targets and multiple links. Network pharmacology reveals effective components，targets and pathway of Tongxinluo capsule in the treatment of cerebral ischemia, which provides theoretical support for the mechanism of Tongxinluo capsule in the treatment of cerebral ischemia.

Research Progress and Application of Mouse Tumor Lung Metastasis Models / 中国实验方剂学杂志

Early diagnoses and treatment methods are being constantly improved, but cancer metastasis remains a main cause of mortality in malignant tumor patients. Lung is thought to be the organ most prone to distal metastasis among malignant tumors due to its unique physiological and pathological character. Tumor lung metastasis is unpredictable and may result in irreversible damages. Presently, no exact mechanism or specific targeting therapies are found. Depending on the unique theory system-treatment based on symptom differentiation, traditional Chinese medicine has made significant progress on controlling tumor lung metastasis, but its application methods and mechanism still need further study and exploration. More appropriate and idealized animal models are required as a studying medium. Therefore, the establishment of animal models to simulate lung metastasis of cancer patients has become the key to the study of tumor lung metastasis. In order to produce a better platform for investigating the pathogenesis, underlying mechanism, early diagnosis and therapeutics for tumor lung metastasis, and to provide reference for the selection and establishment of mouse lung metastasis model, this article would introduce the implementation, application and estimation of several common methods (tail vein injection, mammary fat pad orthotopic injection, tibia injection, tissue orthotopic implantation, transgenic mice and so on). Meanwhile, the development of mouse lung metastasis model still needs expanding of thoughts, rational and flexible utilization of existing models, and interdisciplinary cooperation to establish preferable animal models and make results more reliable.

Research Progress of Different Subtypes of Myeloid-derived Suppressor Cells and Traditional Chinese Medicine Regulation Effect in Tumor Microenvironment / 中国实验方剂学杂志

The diagnosis and treatment methods for cancer are being improved continually, but the mortality of cancer still remains high. At present, the academic circle has realized deficiency of existing treatment ideas, and the concept of cancer cells has been gradually changed from "extremely extinct" to "peaceful coexistence". The concept of "survival with tumors" is universally accepted in the cancer academia. The tumor microenvironment is the place where tumor cells survive and develop. Therefore, regulation of the tumor microenvironment has become an important new strategy for tumor treatment. Myeloid-derived suppressor cells (MDSCs) are a group of heterogeneous cells that have immunosuppressive properties on T cells in the tumor microenvironment and play an important role in tumor immune escape. Now, therapy with MDSCs in the tumor microenvironment as the treatment targets also provides new ideas for the tumor treatment. As MDSCs subpopulations are similar with neutrophils and monocytes, they can be divided into two major subtypes:granulocyte-like myeloid-derived suppressor cells (G-MDSCs) and monocyte-myeloid-derived suppressor cells(M-MDSCs). But how to differ these two subtypes from neutrophils and monocytes. What are the differences in the functional characteristics of different subtypes of MDSCs. How do they accumulate, differentiate, and exert immunosuppressive effects through different pathways. Traditional Chinese medicine(TCM) has always been good at modulating the body's microenvironment. More and more researches have shown that, the recruitment, amplification and activation of MDSCs can be effectively inhibited by TCM compound and its active ingredients, providing scientific basis for Chinese medicine targeting MDSCs in the tumor microenvironment. However, which specific pathways could regulate G-MDSCs or M-MDSCs is still in need of further studies. Most previous literature focus on the overall level of MDSCs, while the this paper would be based on the specific subpopulations of MDSCs to clarify the biological characteristics of these two subtypes of MDSCs, so as to achieve more precise targeted therapy in the tumor microenvironment.

Treatment of hepatic cancer in mice by beta-elemene combined DC/Dribble vaccine: an immune mechanism research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effects of beta-elemene combined DC/Dribble vaccine in treating mice with hepatic cancer, thus exploring their anti-tumor mechanisms.</p><p><b>METHODS</b>Dentritic cells were derived from Balb/c mice's spleen and their phenotypes were identified. Using hepatic cancer cell line BNL1MEA.7R.1 (abbreviated as BNL) originated from Balb/c mice as target cell, DC/Dribble vaccine was prepared via raising the antigen representing carrier autophagosomes (DRips in Blebs, DRibbles), which were rich in tumor antigen information. The mice previously immunized were divided into 4 groups, i.e., the control group, the beta-elemene group, the vaccine group, and the combined group. The PBS was subcutaneously and intraperitoneally injected to mice in the control group. The beta-elemene was intraperitoneally injected at the daily dose of 50 mg/kg to mice in the beta-elemene group and the combined group for 7 successive days. DC/Dribble vaccine was injected into the lymph node of mice in the vaccine group and the combined group on the 1st day, and DC/Dribble vaccine was subcutaneously injected on the 3rd day and the 5th day. All the mice were sacrificed on the 10th day. Their spleens were obtained sterilely, and the suspension was incubated with or without Dribble. The cells were inoculated for 72 h. The contents of IFN-gamma in the supernatant were measured by ELISA. In addition, the spleen cells obtained from the combined group were incubated with different stimulations for 72 h, which were then divided into the control group, the DRibble group, the DC group, and the DC/Dribble vaccine group. The supernatant of cultured cells were collected and the contents of IFN-gamma were measured by ELISA. The liver tumor-bearing mouse model was established, and then the BNL bearing mice were randomly divided into 4 groups, i.e., the control group, the beta-elemene group, the vaccine group, and the combined group. The treatment ways were the same as the immune ways. The tumor size and the survival period were observed in each group. On the 23rd day the mice were sacrificed. The tumor tissue was stripped and stained by HE staining. The pathomorphological manifestations of the tumor tissue were observed by light microscope.</p><p><b>RESULTS</b>In vitro detection of mice immunized previously by different ways showed that the secretion of IFN-gamma was significantly higher in the combined group than in the rest groups (P < 0.01). The secretion of IFN-gamma was significantly higher in the beta-elemene group and the vaccine group than in the control group (P < 0.01). The spleen cells could be stimulated to secrete a large amount of IFN-gamma in the vaccine group and the Dribble group (P < 0.01). When the beta-elemene was 10 microg/mL as the stimulating dose, the secretion of IFN-gamma obviously increased (P < 0.01). In vivo observation showed that the growth velocity of tumors in mice of the combined group was slowed down. There was statistical difference in the tumor area or the survival period of mice in the combined group, when compared with the other groups (P < 0.01). In HE staining, the surrounding connective tissues of the tumor were wrapped tightly and compactedly, with infiltration of a large amount of inflammatory cells.</p><p><b>CONCLUSIONS</b>beta-elemene combined DC/Dribble vaccine could induce specific immune cells to secrete secretory cells, thus exerting its anti-tumor effect. Its immunological effects might be associated with enhancing the DC antigen presenting function.</p>

Effect of integrative medical therapy on the prognosis of patients suffering from yin syndrome type acute ischemic stroke / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of integrative medical therapy on the prognosis of patients suffering from yin syndrome type acute ischemic stroke (YS-AIS).</p><p><b>METHODS</b>A multi-center, prospective, random and parallel controlled clinical trial was carried on 606 patients of YS-AIS. Excepting the 15 patients being excluded and dropped out in the trial period, all patients were assigned to the treatment group (274 patients) treated with integrated medical protocol (i. e. Chinese medical therapy plus conventional Western medical treatment), and the control group (263 patients) treated with conventional Western medical treatment plus placebo. Patients' quality of life (QOL) and their disability level were assessed by scoring based on SS-QOL and modified Rankin scale (mRS) respectively at the terminal of the 21-day treatment (T1) and at the ends of 60-day (T2) and 90-day (T3) following-ups.</p><p><b>RESULTS</b>Patients' disability level, showed by mRS scores, was not significantly different between the two groups at T1 and T2 (P > 0.05), but with significant difference at T3 (P < 0.05). The recovery in the treatment group were more significant. Although the SS-QOL scores showed no statistical significant difference between groups (P > 0.05), improvement of QOL could be seen in the treatment group and the difference between groups was more significant as the time went by.</p><p><b>CONCLUSION</b>Integrative medical therapy could reduce the severe disability rate in patients after YS-AIS, and showed an improving trend on patients' QOL at T3.</p>

Effect of Tongxinluo on nestin and vascular endothehal growth factor mRNA expression in rat brain tissue after cerebral ischemia-reperfusion injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expressions of nestin and vascular endothehal growth factor (VEGF) mRNAs in rat brain tissue after cerebral ischemia-reperfusion injury and their changes in response to Tongxinluo (a traditional Chinese herbal preparation) treatment.</p><p><b>METHODS</b>Cerebral ischemia was induced in rats by temporary middle cerebral artery occlusion (MCAO) followed by treatment with Tongxinluo at high and low doses. On days 3, 7, 14 and 21 after MCAO, nestin and VEGF mRNA expressions in the ependyma, subventricular zone (SVZ), and hippocampal subdentate gyrus zone (HDG) in the ischemic hemisphere were quantitatively analyzed using immunohistochemistry and RT-PCR.</p><p><b>RESULTS</b>Compared with the sham-operated group, the rats with cerebral ischemia-reperfusion injury showed significantly increased nestin-positive neurons and VEGF mRNA expression in the SVZ and HDG 7, 14 and 21 days after MCAO (P<0.05). Treatment with Tongxinluo, especially at high doses, significantly increased the number of nestin-positive neurons and VEGF mRNA expression in the rats 7, 14 and 21 days after MCAO (P<0.05).</p><p><b>CONCLUSION</b>Focal cerebral ischemia in rats results in rapid response and proliferation of neural stem cells in the SVZ and HDG in the ischemic hemisphere possibly by increasing VEGF mRNA expression in the adjacent tissues around the ischemic focus. Tongxinluo may enhance the differentiation and proliferation capacity of the neural stem cells after MCAO by inducing the expression of VEGF mRNA.</p>

Effect of Tongxinluo on the proliferation and differentiation of rat embryonic neural stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Tongxinluo in on the proliferation and differentiation of rat embryonic neural stem cells (NSCs).</p><p><b>METHODS</b>NSCs were isolated from 12- to 14-day SD rat embryo and treated with Tongxinluo at different doses, and the proliferation and differentiation of the cells were observed by immunofluorescence staining at different time points.</p><p><b>RESULTS</b>The ratio of embryonic NSCs labeled with nestin decreased soon after Tongxinluo treatment, but increased afterwards. Significant difference was noted in the number of cells labeled with beta-tubulin between Tongxinluo group and the control group 3 and 7 days after the treatment, and also between high-dose and low-dose Tongxinluo groups at 7 days.</p><p><b>CONCLUSION</b>Tongxinluo can induce the proliferation and neuronal differentiation of rat embryonic NSCs, and the effect is related to the dose of Tongxinluo administered.</p>

Serum hyaluronic acid, tumor necrosis factor -alpha, vascular endothelial growth factor, NO, and Se levels in adult patients with Kashin-Beck disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association of serum levels of hyaluronic acid (HA), tumor necrosis factor-alpha (TNF-alpha), vascular endothelial growth factor (VEGF), NO, and Se with the clinical manifestations in adult patients with Kashin-Beck disease (KBD).</p><p><b>METHODS</b>Total 216 adults were selected for KBD screening from the KBD-prevalent areas in Yongshou county and the non-KBD areas of Chang'an county, Xi'an city, ShaanXi Province. According to the National Diagnostic Criteria of Kashin-Beck Disease in China, the diagnoses of KBD was established in 25 adult patients (11 men and 14 women, average age of 47.88+/-11.16 years), and 20 healthy control subjects from the KBD areas (8 men and 12 women, average age of 47.85+/-12.05 years) and 20 from the non-KBD areas (8 men and 12 women, average age of 47.45+/-11.24 years) were also selected to serve as controls. There was no significant difference in the average age and gender distribution between the 3 groups. The serum levels of HA, TNF-alpha, VEGF, NO and Se were measured by enzyme-linked immunosorbent assay, nitrate reductase method and griphite furnace atomic absorption spectrometry.</p><p><b>RESULTS</b>Serum NO level was significantly higher in KBD group (41.7+/-21.89 micromol/L) than in the health controls from KBD areas (17.1+/-13.01 micromol/L) and non-KBD areas (17.58+/-11.48 micromol/l, F=13.11, df=2, P<0.001). Serum TNF-alpha level in KBD group (32.7+/-3.55 pg/ml) was significantly higher than that in the control subjects from the non-KBD areas (30.95+/-2.22 pg/ml, F=3.672, df=2, P=0.031), but similar with the control subjects from the KBD areas (32.7+/-3.55 pg/ml). Serum TNF-alpha and NO levels were identified as the indices that differed between adult KBD patients and the controls from both KBD and non-KBD areas by differential analysis (the function of differentiation was 0.062xNO+0.173xTNF -7.218).</p><p><b>CONCLUSION</b>Serum TNF-alpha and NO levels are significantly increased in adult KBD patients and are associated with the clinical manifestations of KBD.</p>

Clinical observation on effects of acupuncture at Dazhui (GV 14) for abating fever of common cold / 中国针灸

<p><b>OBJECTIVE</b>To explore the therapeutic effect of acupuncture at Dazhui (GV 14) for abating fever of common cold.</p><p><b>METHODS</b>Two hundred and sixty-one cases were randomly assigned to a treatment group of 133 cases and a control group of 128 cases. The treatment group were treated with electroacupuncture at Dazhui (GV 14) and the control group with antondine injection. The transient effect of abating fever within 24 h was observed.</p><p><b>RESULTS</b>After treatment, the body temperature at all observation time points in the treatment group were lower than those in the control group (P < 0.01). The effect-appearing time (1.42 +/- 1.79) h in the treatment group was shorter than that in the control group (3.44 +/- 5.10) h (P < 0.01). The cured rate and the abating fever rate were 27.8% and 75.9% in the treatment group, and 10.9% and 55.5% in the control group, with significant differences between the two groups, the treatment group being better than the control group (P < 0.01). The abating fever rate for the wind-heat type common cold was 75.3% in the treatment group and 50.0% in the control group, with significant difference between the two groups, the treatment group being better than the control group (P < 0.01).</p><p><b>CONCLUSION</b>The method of acupuncture at Dazhui (GV 14) has a definite therapeutic effect on high fever of common cold, and for wind-heat type common cold, Dazhui (GV 14) first may be chosen to abate high-fever and the treatment should be taken as early as possible.</p>

Nestin activation after rat cerebral ischemia-reperfusion injury and its changes in response to Tongxinluo treatment / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate nestin activation in rat brain subjected to ischemia-reperfusion injury and its changes in response to Tongxinluo treatment.</p><p><b>METHODS</b>Cerebral ischemia was induced by temporary middle cerebral artery occlusion (MCAO) in rats. At 3, 7, 14 and 21 days after MCAO, nestin expression in the ependyma, subventricular zone (SVZ), hippocampal subdentate gyrus zone (HDG) of the rats treated with Tongxinluo were guantified by immunohistochemistry.</p><p><b>RESULTS</b>Compared with the sham operation group, nestin was significantly increased 7, 14 and 21 days after MCAO (P<0.05), and immunofluorescence of BrdU+nestin-positive neurons significantly increased in the SVZ. After treatment with Tongxinluo, the number of BrdU-positive neurons and BrdU+nestin-positive neurons significantly increased as compared with MCAO group (P<0.05).</p><p><b>CONCLUSION</b>Focal cerebral ischemia in the rat results in rapid response and proliferation of neural stem cells in the SVZ and HDG in the ischemic hemisphere, and Tongxinluo may enhance the differentiation and proliferation capacity of the neural stem cells after MCAO.</p>

A new bibenzyl derivative from Bletilla striata / 药学学报

<p><b>AIM</b>To study the chemical constituents of Bletilla striata.</p><p><b>METHODS</b>Various column chromatographies with silica gel and Sephadex LH-20 were employed for the isolation and purification. The structures of the compounds were elucidated on the basis of spectral analyses and chemical methods.</p><p><b>RESULTS</b>Three compounds were isolated from the roots of Bletilla striata (Thunb.) Reichb. f. and identified as 5-hydroxy-4-(p-hydroxybenzyl)-3'-3-dimethoxybibenzyl (I), schizandrin (II), 4,4'-dimethoxy-(1,1'-biphenanthrene)-2,2',7,7'-tetrol (III).</p><p><b>CONCLUSION</b>Compound I is a new bibenzyl derivative and II was isolated from this plant for the first time.</p>