Aim To search for colchicine derivatives which have high efficacy and low toxicity. Methods Colchicine was firstly converted into thiocolchicine, and then it was hydrolyzed to get 7-(N-deacetylthiocolchicine). At last, 7-(N-deacetylthiocolchicine) was amidated to get the target compounds. The chemical structure of these new derivatives was confirmed with 1H NMR, IR, MS, and HR-MS. The cytotoxicity of the compounds was tested by MTT assay. Their in vivo antitumor activity was evaluated against mice tumor H22 and U14. Results Twelve thiocolchicine derivatives are new compounds. Conclusion In vitro antitumor activity has showed that some of these thiocolchicines possessed cytotoxic activity superior to colchicine. However, in vivo antitumor activity indicated that these derivatives have poor efficacy in mice.

Aim To investigate the effects of cyclosporin A (CsA) on growth and collagen synthesis of cardiac fibroblasts (CFs) induced by arginine vasopressin (AVP). Methods CFs of neonatal Sprague-Dawley rats were isolated by trypsinization and cultured; growth-arrested CFs were stimulated with 1×10-7 mol·L-1 AVP in the presence or absence of CsA (0.05, 0.5 and 5 μmol·L-1). MTT and flow cytometry techniques were adopted to measure cell number and analyze cell cycle respectively. Collagen synthesis was determined by measurement of hydroxyproline content in culture supernatant with colorimetry. Calcineurin activity was estimated by chemiluminescence. Trypan blue staining to test the viability of CFs. Results 0.05, 0.5 and 5 μmol·L-1 CsA inhibited the increase of CFs number induced by 1×10-7 mol·L-1 AVP in a dose-dependent manner, with the inhibitory rates by 12%, 24% and 29%, respectively (P＜0.05). Furthermore, cell cycle analysis showed 0.5 μmol·L-1 CsA decreased the S stage percentage and proliferation index of CFs stimulated by AVP (P＜0.05). In culture medium, the hydroxyproline content induced by AVP decreased by 0.5 and 5 μmol·L-1 CsA (P＜0.05), with the inhibitory rates of 29% and 33%, respectively. CsA completely inhibited the increment of calcineurin activity induced by AVP (P＜0.01), but CsA itself had no effect on the baseline of calcineurin activity and CFs viability. Conclusion CsA inhibits proliferation and collagen synthesis of CFs by virtue of blocking calcineurin signaling pathway and might provide a novel target for prevention and treatment to cardiac fibrosis.

Aim To explore the modulation of Salvia miltiorrhiza on hyperpolarization-activated current (Ih) channels in dorsal root ganglion (DRG) neurons of rats and identify the mechanism of Salvia miltiorrhiza in alleviating pain and inhibiting calcium overload. Methods The effect of Salvia miltiorrhiza injection on Ih channels in DRG neurons of rats were examined by using whole-cell patch clamp technique. Results The experimental results showed that the amplitude of Ih evoked by -150 mV was (-1.06±0.18) nA. The Ih could be fitted well into the single kinetics and the time constant of activation, τ was clearly voltage-dependent with τ=(322.14±28.81) ms at -100 mV, decreasing to τ=(62.51±9.78) ms at -150 mV. The reversal potential of Ih was (-35.03±1.12) mV measured from tail currents. But no significant differences were found between the DRG neurons in the absence and presence of Salvia miltiorrhiza injection (10%, 25%, 50%) in the current amplitude, the time constant of activation and the reversal potential. The only difference between the DRG neurons in the absence and presence of Salvia miltiorrhiza injection was the half-activation potential of Ih. In control recordings the half-activation potential was (-106.07±3.59) mV. By comparison, the half-activation potentials changed to (-111.59±3.79) mV (n=31 neurons, P＜0.05), (-119.37±4.96) mV (n=31 neurons, P＜0.05) and (-121.23±3.86) mV (n=31 neurons, P＜0.05) in the presence of 10%, 25%, 50% Salvia miltiorrhiza injection, respectively. Conclusion Only the half-activation potential of Ih in the arthritic and neuropathic rat models shifted in the depolarizing direction, which increased the electrophysiological activity of Ih and made it related to peripheral hyperalgesia. The selective inhibition of Salvia miltiorrhiza on the electrophysiological activity of Ih may be one of the mechanisms underlying its analgesic effects.

Aim To investigate the water-soluble phenolic glycosides from the whole plant of Bulbophyllum odoratissimum. Methods Column chromatography techniques were used to isolate the chemical constituents, physico-chemical constants and spectroscopic analysis were employed for structural elucidation. Results Bulbophyllinoside (1), a new phenolic glycoside and three known compounds were isolated from the whole plant of Bulbophyllum odoratissimum Lindl. Their structures were determined as 3-hydroxyphenethyl alcohol 4-O-(6'-O-β-apiofuranosyl)-β-D-glucopyranoside (1), 3-methoxyphenethyl alcohol 4-O-β-D-glucopynanoside (2), 3,5-dimethoxyphenethyl alcohol 4-O-β-D-glucopynanoside (3) and syringin (4). Conclusion Bulbophyllinoside (1) is a new compound.

Aim To study the chemical constituents of Ardisia punctata. Methods Compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and single crystal X-ray diffraction. Results Three compounds were isolated from chloroform extract of the roots of Ardisia punctata. Their structures were elucidated as 2-tridecyl-3-[ (2-tridecyl-3-acetoxy-4-methoxy-6-hydroxy) -phenyl ] -6-methoxy-1,4-benzoquinone ( 1 ), 2-tridecyl-3-[ ( 2 -tridecyl-4,6-dihydroxy ) -phenyl ] -6 -methoxy-1,4-benzoquinone ( 2 ) and 2 -tridecyl-3 - [ ( 2 -pentadecyl-4,6-dihydroxyl)-phenyl]-6-methoxy-1,4-benzoquinone (3). Conclusion The three compounds are new1,4-benzoquinone derivatives.

Aim To investigate apoptosis induced by 3,3'-diethyl-9-methylthia-carbocyanine iodide(DMTCCI) , an inhibitor of DNA primase found in our previous study, and the mechanism of DMTCCI in human myelogenous leukemia HL-60 cells. Methods HL-60 cells were cultured in RPMI-1640 medium and treated with different concentrations of DMTCCI. MTT assay was used to detect growth inhibition.Flow cytometry and DNA ladders were used to detect apoptosis. Western blotting was used to observe the expression of survivin, Bcl-xL, Bad, Bax, Bcl-2, caspase-9, caspase-3, caspase-6, PARP, DFF45 and lamin B protein. Caspase-3 activity was measured by ApoAlert Caspase-3 Assay Kit. Results DMTCCI inhibited proliferation of human leukemia HL-60 cells with IC50 value of 0. 24 μmol · L-1. The results of flow cytometry and DNA ladders showed that DMTCCI could induce apoptosis of HL-60 cells. The expression levels of protein survivin and Bcl-xL were down-regulated, Bad and Bax were up-regulated,while Bcl-2 protein had no change in response to DMTCCI treatment in HL-60 cells. Treatment of HL-60cells with DMTCCI induced the proteolytic cleavage of caspase-9, caspase-3, caspase-6, PARP, DFF45and lamin B protein. Caspase-3 activity apparently increased at 3 h and reached a peak at 12 h after exposure to 1 μmol · L-1 of DMTCCI in HL-60 cells. Conclusion DMTCCI inhibited proliferation and induced apoptosis of human leukemia HL-60 cells. Bcl-2 family proteins, survivin and caspases family proteins might playa role in the apoptosis process induced by DMTCCI.

Aim To prepare a new oral colon-specific delivery formulation and to investigate the release profile in vitro and the colon-specific delivery property in vivo in dogs. Methods Sodium 4-aminosalicylic acid was selected as the model drug. The combination of Eudragit RL30D and RS30D were used as sustained-release film, and Eudragit FS30D used as enteric film, which was expected to release drug depending on pH and time. The release profile of tablets was studied in three phosphate buffers with the pH 6.5, 7.0 or 7.4 for 12 h after a simulated gastric presoak for 2 h in 0.1 mol · L-1 HCl. The tablets were radiolabelled with 99mTc to make their release times and positions in the gastrointestinal tract be followed using a gamma camera. Results For the in vitro study, there was no drug released in 0. 1 mol ·L- 1 HCl for 2 h, and release occurred slowly when pH was above 6.5. Drug was released faster while pH was higher. For the in vivo study, the coated tablets remained intact in the upper gastrointestinal tract, and drug release began after the colonic arrival. The uncoated tablets, however, disintegrated in the stomach of the dogs rapidly. Conclusion The coating could protect the drug until the tablets reached the ascending colon, where drug was released slowly for over 10 h.

Aim To investigate the induction of endothelial cell apoptosis and the suppression of VEGF expression in cancer cells by sodium caffeate (SCA). Methods Apoptosis of transformed human umbilical vein endothelial cells (ECV304 cell line) was detected by flow cytometry, DNA electrophoresis assay and morphological assessment. Western blotting analysis was applied for determination of VEGF expression in cancer cells. Substrate degradation by type Ⅳ collagenase was measured by zymography.ELISA was used to detect the binding of type Ⅳ collagenase with relevant monoclonal antibody. Results SCA induced ECV304 cell apoptosis in a time- and dose-dependent manner. After treatment with 100 and fluorescence and distinct changes of nuclear morphology, such as pyknosis and the occurrence of apoptotic bodies. VEGF expression in hepatoma HepG-2 cells and prostate carcinoma DU145 cells was reduced after SCA treatment. The degradation activity of type Ⅳ collagenase including MMP-2 and MMP-9 secreted by giant cell pulmonary carcinoma PG cells was inhibited by SCA in a dose-dependent manner. SCA also reduced the binding of mAb 3D6, a relevant monoclonal antibody, to type Ⅳ collagenase. Conclusion SCA can induce endothelial cell apoptosis and inhibit VEGF expression as well as type Ⅳ collagenase activity in cancer cells. SCA might be active in modulating tumor angiogenesis and the microenvironment.

Aim To study the effect of ZD7288 on synaptic transmission in the pathway from perforant pathway (PP) fibers to CA3 region in rat hippocampus. Methods The extracellular recording technique in vivo was used to record the CA3 region field potentials. High-performance liquid chromatography (HPLC) with fluorescence detection was applied to measure the content of amino acids in hippocampal tissues. The effect of ZD7288 and CsCl on the amplitudes of population spike (PS) in CA3 region evoked by stimulation (0.5 Hz) of the perforant pathway (PP) fibers, and the content of amino acids in hippocampal tissue were observed. Results Microinjection of ZD7288 (20, 100 and 200 nmol)and CsCl (1,5 and 10 μmol) into CA3 region decreased the population spike (PS) amplitudes in a dosedependent manner. The inhibitory effects appeared at 5 min after microinjection and lasted at least 90 min.In those rats treated with ZD7288 (100 nmol), the contents of glutamate, aspartate, glycine and GABA decreased significantly as compared to those of saline control ( all P＜0.01, except P＜0.05 for that of glycine). A similar decrease in the contents of amino acids was observed when the rats were microinjected with CsCl (5 μmol). Conclusion ZD7288 could obviously inhibit synaptic transmission in the pathway from PP fibers to CA3 region in rat hippocampus, and this action of ZD7288 may be associated with altered contents of amino acids.

Aim To evaluate the effect of paeonol on the activity of tyrosinase and provide experimental evidence for the treatment of hyperpigmentation disorders. Methods Tyrosinase activity was estimated by measuring the oxidation rate of L-3,4-dihydroxyphenylalanine (L-Dopa). The inhibitory effects of paeonol on the activity of mushroom tyrosinase and Michaelis-Menten kinetics were deduced from the Lineweaver-Burk plots. Results The inhibitory concentration of paeonol leading to 50% enzyme Paeonol is a potential mixed inhibitor of mushroom tyrosinase. The mixed inhibition function may originate from its ability to form a Schiff base with a primary amino group and to chelate copper at the active site of tyrosinase.