1.Optimization of induction and culture conditions for hairy roots of Salvia miltiorrhiza.
Rong-Hui TAN ; Jin-Jia ZHANG ; Shu-Juan ZHAO
China Journal of Chinese Materia Medica 2014;39(16):3048-3053
To establish induction and liquid culture system for hairy roots of Danshen (Salvia miltiorrhiza), Agrobacterium rhizogenes A4, LBA9402, 15834 as test bacterium were used to infect aseptic leaves of Danshen. The hairy roots were induced and positive transgenic hairy roots were selected with PCR using rolB and rolC as the target gene. Then hairy roots of S. miltiorrhiza were harvested and salvianolic acids were extracted with 70% methanol containing 1% formic acid. The content of salvianolic acid B (SalB) and rosmarinic acid (RA) were determined by HPLC. According to the above research results, the Danshen hairy roots induced by A. rhizogenes LBA9402 were inoculated into the following group of culture media: MSOH, MS, B5, and 6,7-V liquid media. Then the same methods of extraction and determination for the content of Danshen hairy roots were adopted. Last, the hairy roots of S. miltiorrhiza induced by A. rhizogenes LBA9402 were inoculated into the MSOH liquid media with different pH values. The content of salvianolic acid were extracted with 70% methanol containing 1% formic acid and determined by HPLC. As a result, three kinds of A. rhizogenes A4, LBA9402, 15834 could induce hairy roots and Ri plasmids were integrated into the genome of S. miltiorrhiza by PCR. Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid, which were (3.27 ± 0.37)% [including (1.04 ±0.36)% of RA and (2.22 ± 0.29)% of SalB] and (3.17 ± 0.20)% [including (0.92 ± 0.31)% of RA and (2.25 ± 0.26)% of SalB], respectively. Hairy roots induced by A. rhizogenes LBA9402 when they were cultured in MSOH liquid media produced much more salvianolic acid, which was (4.56 ± 0.36)%, including (1.12 ± 0.26)% of RA and (3.44 ± 0.23)% of SalB. Hairy roots induced by A. rhizogenes LBA9402 produced the most salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81, which was 4.85%, including 1.16% of RA and 3.69% of SalB. So Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81. The research had established the foundation on genetic engineering to improve the quality of S. miltiorrhiza.
Agrobacterium
;
physiology
;
Benzofurans
;
analysis
;
metabolism
;
Cell Culture Techniques
;
instrumentation
;
methods
;
Cinnamates
;
analysis
;
metabolism
;
Culture Media
;
chemistry
;
metabolism
;
Depsides
;
analysis
;
metabolism
;
Drugs, Chinese Herbal
;
analysis
;
metabolism
;
Plant Roots
;
chemistry
;
growth & development
;
metabolism
;
microbiology
;
Salvia miltiorrhiza
;
chemistry
;
growth & development
;
metabolism
;
microbiology
2.Photo-toxic effects of indocyanine green based on photodynamic therapy on vascular endothelial cells in ;vitro
Shaobo ZHOU ; Haifeng HONG ; Juan TAN ; Hui LI
The Journal of Practical Medicine 2015;(6):908-911
Objective To investigate the photo-toxic effects of indocyanine green (ICG) based on photodynamic therapy (PDT) on vascular endothelial cells in vitro. Methods Human umbilical vein endothelial cells were incubated with ICG with different concentrations in 96-well plate. According to the dose of ICG , they were divided into 6 groups as 0, 50, 100, 150, 200 and 250 μmol/L group. Each group consists of 8 wells, 4 of them were irradiated with a 810 nm laser of total power 24.0 J/cm2. Cell viability was measured by MTT assay 24 h after light irradiation. Morphological changes in cells were observed by light microscope. Results With the irradiation of 810 nm light , there was a significant decrease in relative cell viability in all groups except 0 μmol/L group (all P < 0). With the increase of concentration of ICG, the cell viability dropped rapidly (P < 0). Apoptotic changes can be seen in all groups except 0 μmol/L group and necrosis was the main change in cells of 200 μmol/L and 250 μmol/L group. Conclusion PDT with appropriate concentration of ICG has a strong inhibition to vascular endothelial cells in vitro and ICG-PDT may be a good new therapy for nevascularization.
3.Effect of behavior training on the expression of NR2B around the infarcted focus and in the cortex of temporal lobe in rats with bilateral hippocampal infarction
Hui-juan PAN ; Ling LI ; Hua YANG ; Shan JIANG ; Yongxia TAN
Chinese Journal of Rehabilitation Theory and Practice 2006;12(1):5-7
ObjectiveTo study the effect of behavior training on the expression of NR2B around the infarcted focus and in the cortex of temporal lobe in rats with bilateral hippocampal infarction.Methods54 SD rats were randomized into pre-training, 7 d, 14 d, 21 d after training groups, pre-immobilization, 7 d, 14 d, 21 d immobilization groups and control group. Behavior training and immobilization were performed on the 3rd day after the infarction. Immunohistochemistry was used to detect the expression of NR2B around the infarcted focus and in the cortex of temporal lobe at different points of time.ResultsThe expression of NR2B was abundant in the normal hippocampus and cortex of temporal lobe of rats. The expression of NR2B decreased after infarction and increased after behavior training. However, the expression of NR2B had little increased in immobilization groups, and showed significant difference compared with that in behavior training groups (P<0.01).ConclusionBehavior training can accelerate the expression of NR2B around the infarcted focus and in the cortex of temporal lobe in rats with bilateral hippocampal infarction.
4.SmHPPR1 from Salvia miltiorrhiza regulated the biosynthesis of salvianolic acids
Rong-hui TAN ; Wang ZHAO ; Jin-jia ZHANG ; Shu-juan ZHAO
Acta Pharmaceutica Sinica 2023;58(9):2818-2828
italic>Salvia miltiorrhiza Bunge is a traditional Chinese medicinal herb widely used to treat cardiovascular and cerebrovascular diseases at clinic. Its main water-soluble components are rosmarinic acid (RA) and salvianolic acid B (SAB), which are produced by phenylpropanoid pathway. 4-Hydroxyphenylpyruvate reductase (HPPR) is a key enzyme in phenylpropanoid metabolism pathway.
5.Effects of Lonicera Japonica flavone on immunomodulation in mice.
Jian-hui PI ; Juan TAN ; Zhao-tun HU ; De-biao XIANG
Chinese Journal of Applied Physiology 2015;31(1):89-92
OBJECTIVETo study immunomodulating activity of Lonicera Japonica flavone by investigating immune enzymatic activity of serum and antoxidized activity of lymphoid organs in mice.
METHODSFifty KM mice were randomly divided into control group, model group, low dose group, middle dose group and high dose group(n = 10), respectively. And low dose group, middle dose group and high dose group were given Lonicera Japonica flavone with 100 mg/kg, 200 mg/kg and 400 mg/kg every day, respectively, while control group and model group were administered with NS. After continuously giving drug 7 weeks, other groups were injected with Dexamethasome (Dex: 25 mg /kg) for 3 days by subcutaneous injection, but the control group were treated with NS. And after giving Lonicera Japonica flavone 1 week simultaneously, organ indexes , the activity of acid phosphatase (ACP), alkaline phosphatase (AKP) and lysozyme (LSZ) in serum , and the content of monoamine oxidase (MAO), total antioxidant capacity (T-AOC), total superoxide dismutase (SOD) and malondialdehyde (MDA) in lymphoid organs in mice were tested, respectively.
RESULTSLonicera Japonica flavone could significantly improve the organ indexes, and significantly improve the activity of ACP, AKP and LSZ in serum, and significantly improve the contents of T-AOC and SOD, but reduce that of MAO and MDA in lymphoid organs in immunosuppressed mice.
CONCLUSIONIonicera Japonica flavone can significantly improve the activity of immune enzyme in serum and the antioxidized activity of lymphoid organs in mice. It suggests that Ionicera Japonica flavone has a good immunomodulatory effects.
Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Antioxidants ; metabolism ; Flavones ; pharmacology ; Immunomodulation ; Lonicera ; chemistry ; Malondialdehyde ; metabolism ; Mice ; Monoamine Oxidase ; metabolism ; Muramidase ; blood ; Superoxide Dismutase ; metabolism
6.Treatment of Epidermal Growth Factor Receptor Inhibitors Associated Adverse Skin Reactions by Zhiyang Pingfu Liquid: a Clinical Study.
Hong-yan WANG ; Chao ZOU ; Hui-juan CUI ; Yan-ping BAI ; Yuan LI ; Huang-ying TAN ; Wei WANG ; Hai JU
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(7):820-822
OBJECTIVETo study the curative effect of Zhiyang Pingfu Liquid (ZPL) in treating epidermal growth factor receptor inhibitors (EGFRIs) associated adverse reactions of the skin.
METHODSAll 54 patients with pathologically confirmed malignant tumor had EGFRIs induced adverse reactions of the skin to various degrees. ZPL was externally applied for them all, once or twice per day, 14 days consisting of one therapeutic course. Changes of adverse skin reactions, time for symptoms relief, adverse skin reaction types suitable for ZPL were observed before and after treatment.
RESULTSEGFRIs associated skin adverse reactions were improved to various degrees after they used ZPL. The shortest symptoms relief time was 1 day while the longest was 12 days, with an average of 6.93 days and the median time 7 days. Compared with before treatment, itching, rash/scaling, acne/acneform eruptions were obviously improved (P < 0.05).
CONCLUSIONZPL could alleviate EGFRls associated adverse skin reactions, especially showed better effect on itching, rash/scaling, acne/acneform eruptions.
Antineoplastic Agents ; adverse effects ; Biomedical Research ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Exanthema ; chemically induced ; Humans ; Neoplasms ; drug therapy ; Pruritus ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; Skin ; drug effects ; Skin Diseases ; drug therapy
7.Auricular acupuncture for primary insomnia: a systematic review based on GRADE system.
Hui-Juan TAN ; Ying LAN ; Fu-Sheng WU ; Han-Dan ZHANG ; Li WU ; Xi WU ; Fan-Rong LIANG
Chinese Acupuncture & Moxibustion 2014;34(7):726-730
The clinical efficacy and safety of auricular acupuncture (AA) for treatment of primary insomnia was evaluated. After a comprehensive retrieval in domestic and foreign databases, literatures were strictly screened and Revman 5.2 software was applied to perform a Meta-analysis on eligible randomized controlled trials (RCTs). The evidence quality was assessed with GRADE profiler 3.6 software. As a result, 8 articles were included involving 894 patients. Compared among AA and sham AA, placebo AA, blank control, there was significant difference in Pittsburgh sleep quality index (PSQI) [WMD = -3.48, 95% CI (-3.96, -3.00)], sleep latency LWMD = -10.14, 95% CI (-17.16, -3.12)] and sleep awakening times [WMD = -9.98, 95% CI (-1.10,-0.48)]. Compared between AA and western medication, there was significant difference in PSQI [WMD = -3.62, 95% CI (-4.59, -2.65)]. The evidence quality was moderate in AA vs. sham AA, placebo AA or blank control, while that of the rest was extremely low. No reports of adverse events were described in all studies. In conclusion, for the treatment of primary insomnia, AA could effectively improve sleep quality, but due to the low evidence quality, cautious attitude should be taken on this conclusion, and clinical trials with large sample and high quality were needed in the further.
Acupuncture, Ear
;
Humans
;
Randomized Controlled Trials as Topic
;
Sleep
;
Sleep Initiation and Maintenance Disorders
;
physiopathology
;
therapy
8.Localization and differentiation of hair follicle stem cells.
Song-Mei GENG ; Jian-Li WANG ; Wan-Juan WANG ; Sheng-Shun TAN ; Zhen-Hui PENG
Acta Academiae Medicinae Sinicae 2006;28(3):360-363
OBJECTIVETo identify the localization of hair follicles stem cell (HFSC) in different stages of hair and explore the differentiating capacity of HFSC into epidermis in vitro.
METHODSHFSC were detected by K19 immunostaining in normal human skin. Then, the isolated HFSC through enzyme digestion were seeded on dermal equivalent (DE) and cultured between the air-liquid interfaces for 14 days. Skin-equivalents was harvested and used for evaluation.
RESULTSHFSC mainly located in outer root sheet in hair follicle and human anagen hair follicles containing two distinct reservoirs for K19-positive cells located in the bulge and bulb of the follicle. These two reservoirs fused in line of outer root sheets during the catagen-telogen transition phase and individualized again in the newly forming anagen hair follicle. Based on DE, growing HFSC built a multilayered and confined epidermis.
CONCLUSIONHFSC located in outer root sheets can promote hair cycle and differentiate into epidermis in vitro.
Cell Differentiation ; physiology ; Cells, Cultured ; Epidermis ; cytology ; Hair Follicle ; cytology ; Humans ; Stem Cells ; cytology
9.Effects of ternary complexes of copper with salicylaldehyde-amino acid Schiff base coordination compounds on the proliferation of BGC823 cells.
Ai-Juan GUO ; Xiao-Shan XU ; Ying-Hui HU ; Ming-Zhao WANG ; Xin TAN
Chinese Journal of Cancer 2010;29(3):277-282
BACKGROUND AND OBJECTIVEThe basic structure of salicylaldehyde-amino acid Schiff base compounds includes a C=N chemical bond. These compounds show significant antitumor activities in vitro when combined with a metal ion. This study investigated the effects and possible mechanisms of four salicylaldehyde-amino acid Schiff base copper ternary coordination compounds on the proliferation of human gastric cancer cell line BGC823.
METHODSThe BGC823 cells were treated with the four compounds (6B, 7B, 6P, and 7P). Cell proliferation was detected by MTT assay. Apoptosis and changes in the cell cycle were analyzed by flow cytometry. DNA damage was observed using a DNA ladder assay. The expression of p53 protein was determined by immunocytochemistry.
RESULTSThe proliferation of BGC823 cells was significantly inhibited by the four compounds and the effect was concentration-dependent. The half maximal inhibitory concentration (IC50) of 6B, 7B, 6P, and 7P for BGC823 cells were 18.10, 27.50, 3.61, and 3.45 micromol/L, respectively. Flow cytometry showed the four drugs induced apoptosis in BGC823 cells, which was confirmed by DNA ladder experiments. Flow cytometry also detected changed phases in the cell cycle from treatment with the compounds. The percent of cells in the G(0)/G(1) phase decreased and that of cells in the G1/S and G(2)/M phases increased, indicating that S-and G2-phase blockages exist. As shown by immunocytochemistry, the expression of p53 decreased in BGC823 cells treated with the four drugs, indicating the involvement of the p53 pathway to BGC823 cell apoptosis.
CONCLUSIONSThe four compounds showed significant activities on restraining proliferation of BGC823 cells in vitro, induced apoptosis, and caused changes in the cell cycle. This may be related to the downregulation of p53.
Aldehydes ; chemistry ; Amino Acids ; chemistry ; Antineoplastic Agents ; chemical synthesis ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Coordination Complexes ; chemical synthesis ; pharmacology ; Copper ; chemistry ; Humans ; Inhibitory Concentration 50 ; Schiff Bases ; chemistry ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism
10.Enhancement of tropane alkaloids biosynthesis in Atropa belladonna hariy root by overexpression of HnCYP82M3 and DsTRI genes
De-hui MU ; Yan-hong LIU ; Piao-piao CHEN ; Ai-juan TAN ; Bing-nan MA ; Hang PAN ; Ming-sheng ZHANG ; Wei QIANG
Acta Pharmaceutica Sinica 2024;59(3):775-783
Tropane alkaloids (TAs) are a class of anticholinergic drugs widely used in clinical practice and mainly extracted from plant, among which