Objective To study the chemical constituents in Crepis turczaniowii C. A. Mey. Method The constituents were isolated and purified by column silica,polystyrene resin RA and polyamide columbine chromatography,and the structures were identified by physicochemical data,MS and NMR. Result Two compounds were obtained in the Petroleum ether fractions as ?-taraxasteryl acetate (Ⅰ) and ?-sitosterol (Ⅱ),and one compound was obtained in the n-BuOH fractions as phillyrin (Ⅲ). Conclusion All above compounds are obtained from the plants of Crepis turczaniowii C. A. Mey. for the first time. Ⅲ is isolated from Crepis L. for the first time.

Object To examine if Leonurus artemisia (Lour ) S Y Hu (Labiatae) contains ferulic acid Methods The samples were extracted by different solvents and the extracts examined by TLC and HPLC Results Ferulic acid was detectable in the extracts of L artemisia Conclusion L artemisia possibly contains free ferulic acid The use of 5% Na 2CO 3 as the extractant instead of methanol, as described in Chinese Pharmacopoeia seemed to be more simple, highly stable and with good reproducibility

Objective: To establish the quality standard for Qingchunle Capsules. Methods: Paeoniflorin of Qingchunle Capsules was determined by dual wave length TLC scanning, ? S=230nm, ? R=320nm. Radix salviae Miltiorrhizae, Radix Angelicae Sinennsis and Radix Ginseng Rubra were identified by TLC.Results: The average recovery rate was up to 97.9% and RSD was 3.0%. The TLC sports developed were fairly clear, and the blank test showed no interference. Conclusion: This method is reliable. The results are stable with good reproducibility. This method can be used for quality control of the capsules.

Objective: To compare supercritical CO 2 fluid extraction(SFE CO 2) with steam distillation(SD) in the chemical constituents and content of essential oil from Forsythia Suspensa. Method: GC MS combination was adopted. Results: Both the two methods shared the same main components as ? pinene, ? pinene and sabinene. Benzene methanol,4 [1 methylethyl] and Benzene methanol,2 [4 methylethyl] were extracted exclusively by SFE CO 2, Among two methods there is a good deal of similarity all the other components extracted. Furthermore, sabinene、transpinocarveol and shorten were the first obtained from the plant. Conclusion: The SFE CO 2 was superior to SD in raising yield and reducing extractive time. It is a good method in extracting the essential oils from Forsythia Suspensa.

Objective To establish the quality standard for Sanyuan Rupixiao Gel Paste. Methods Sparganii Rhizoma, Gleditsiae Sinensis Fructus, Cyperi Rhizoma and Impatientis Semen were identified by TLC method. The content of tetrahydropalmatine was determined by HPLC. Waters symmetry column was used with the mobile phase of acetonitrile-0.1% phosphatic acid in a gradient manner (pH was adjusted to 6.4 by triethylamine) (55:45) at the detection wavelength of 280 nm. The flow rate was 1.0 mL/min at the column temperature of 30 ℃. Results The spots in TLC were clear without any interference;tetrahydropalmatine showed a good linear relation in the range of 0.092–1.84 μg;the average recovery was 100.15%with RSD of 1.58%(n=6). Conclusion The method is simple and accurate with high reproducibility, which can be used for the quality control of Sanyuan Rupixiao Gel Paste.

OBJECTIVETo optimize the extraction technology and determine the immune activity of polysaccharides from Marsdenia tenacissima.

METHODThe optimum extraction technology of polysaccharide from M. tenacissima was detected by orthogonal experiments with the extraction rate of polysacchride and the total sugar content as indicators. The immunocompromised mice model was established by intraperitoneal injection cyclophosphamide to detected the content of IL-2, IL-6 in serum, CD4+, CD8+ in the peripheral blood by ELISA and flow cytometry, respectively.

RESULTBy the extraction rate of polysaccharide, the sequence of seriousness of all affecting factors from high to low was extracting times, temperature, heating time and water ratio. By the total sugar content, the sequence was temperature, extracting times, water ratio and heating time. Compared with the model group, the pleen index, IL-2, IL-6, CD4/CD8+ were increased significantly (P < 0.05) in the 0.14 g x kg(-1) group and 0. 28 g x kg(-1) group.

CONCLUSIONT he optimum extraction condition was as follows: extraction three times/1.5 hours at 100 degrees C with 1:8 ratio of M. tenacissima to water. The polysaccharide of M. tenacissima can enhance the cellular immune and humoral immune.

Animals ; Body Weight ; drug effects ; CD4-CD8 Ratio ; Chemical Fractionation ; methods ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Marsdenia ; chemistry ; Mice ; Polysaccharides ; immunology ; isolation & purification ; pharmacology ; Spleen ; immunology ; Temperature ; Time Factors

Objective To investigate the proliferative inhibition effect and mechanism of total saponins from marsdenia tenacissima on human liver cancer HepG2 cells. Methods Human liver cancer HpeG2 cells cultured conventionally were divided into the marsdenia tenacissima saponins A control group, the experimental group and the blank control group. The experimental group was treated with different mass concentrations of total saponins from marsdenia tenacissima (0.14, 0.29, 0.58, 1.15, 2.13 mg/ml), while the marsdenia tenacissima saponins A control group was treated with marsdenia tenacissima saponins A (1.0 mg/ml), and the blank control group was established stimultaneously. The methyl thiazolyl tetrazolium (MTT) method was used to detect the effect of total saponins from marsdenia tenacissima on the activity of HepG2 cells, and the cell morphology was observed by using inverted microscopy. The cell apoptosis rate was detected by using flow cytometry from Annexin V-FITC/PI staining. The expressions of bcl-2, bax and p53 were analyzed by using Western blot after the drug effect. Results The results of cell activity test showed that total saponins from marsdenia tenacissima could inhibit the proliferation of HepG2 cells at the concentration of 0.29, 0.58, 1.15, 2.13 mg/ml, which was positively correlated with the concentration. The half maximal inhibitory concentration (IC 50) of cell growth inhibition of total saponins from marsdenia tenacissima on HepG2 cells was 0.75 mg/ml. Under inverted microscopy, the adherent cells were significantly reduced, the cells fell off into clusters and the debris was increased after the effect of total saponins from marsdenia tenacissima. Moreover, flow cytometry showed that total saponins from marsdenia tenacissima could increase the late apoptosis rate of HepG2 cells (P< 0.01). Western blot showed that the expression of bcl-2 was 0.62±0.16, 0.31±0.15, 0.84±0.09 and 1.00±0.11 respectively in the experimental group (total saponins from marsdenia tenacissima 0.58 mg/ml and 2.13 mg/ml), the marsdenia tenacissima saponins A control group and the blank control group;the expression of bax protein was 0.75±0.10, 0.83±0.12, 1.00±0.14 and 0.15±0.02, respectively in the above four groups; the expression of p53 protein was 0.63±0.08, 0.78±0.11, 1.00±0.13 and 0.18±0.02 respectively in the above four groups. The protein expression of bcl-2 was decreased and the protein expressions of bax and p53 were increased in the experiment group, and there was a statistical difference between the experiment group and the blank control group (P< 0.05). Conclusions Total saponins from marsdenia tenacissima can upregulate the expression of bax by upregulating the expression of p53 gene, and inhibit the expression of bcl-2, which would cause the cascade reaction to induce the apoptosis of HepG2 cells. Its inhibitory effect can be realized through mitochondrial pathmay to induce apoptosis.

With the appearance of the disadvantages of traditional tumor treatment, immunotherapy has entered people′s horizons as modern emerging treatment strategies, among which plant polysaccharides have received much more attention due to their antitumor activity and significant immunomodulatory effects. Tumor-associated macrophages (TAMs), as a component of tumor microenvironment, are important factors affecting tumors, and the regulation of TAMs by plant polysaccharides is one of the effective immunotherapy to treat tumor. In this review, we mainly described the regulation of TAMs by plant polysaccharides and the underlying mechanisms, and then gave an outlook on the research interests and the development of plant polysaccharides as immune adjuvants, aiming to provide reference for the study of plant polysaccharides in the immunotherapy for tumors.

Macrophages are important cells of the immune system. Tumor-associated macrophages are enriched macrophages near tumor cells or tissues. Their role is mainly to promote the construction of tumor inflammatory microenvironment and inhibit tumor immune response. Cell co-culture system is a symbiotic culture system formed by mimicking the internal environment of the body in vitro. The co-culture condition is relatively consistent with the environment in vivo, enabling better information exchange and material exchange between cells, which is a supplement to the monolayer cell culture and animal experiments. Tumor-associated macrophages and tumor cells co-exist in the tumor microenvironment. Thus, constructing a co-culture system for tumor-associated macrophages and tumor cells would be conducive to studying the antitumor effect of tumor-associated macrophages and developing new immunotherapy drugs. The co-culture system would provide a new direction for treating malignant tumors. This article mainly reviewed the co-culture patterns of macrophages and the antitumor effects of different phenotypes of macrophages, and highlighted the importance of using immunotherapy to treat malignant tumors in the tumor microenvironment.