Phenolic Compounds from the Rhizomes of Smilax china L. and Their Anti-Inflammatory Activity.

A new triflavanoid, kandelin B-5 (1), was isolated from the rhizomes of Smilax china L., together with six known phenylpropanoid substituted flavan-3-ols (2-7), nine flavonoids (8-16), two stilbenoids (17, 18), and two other compounds (19, 20). The structure of compound 1 was determined on the basis of 1D, 2D NMR and HR-ESI-MS data, as well as chemical method. Compounds 2-5, 8-12, 15, 17, and 19 were evaluated for anti-inflammatory activity. Only compounds 10, 15 and 17 showed slightly IL-1ß expression inhibitory activities on LPS induced THP-1 cells, with inhibition rate of 15.8%, 37.3%, and 35.8%, respectively, at concentration of 50 µg/mL.

[Preparation and in vitro drug release study of long-circulating hydroxycamptothecin nanoparticles].

OBJECTIVE: To prepare long-circulating hydroxycamptothecin nanoparticles and study its in vitro drug release characteristics. METHODS: The HCPT-PEG-PCL-NPs were prepared by solvent-diffusion method using PEG-PCL block copolymer synthesized as a matrix and HCPT as an antitumor agent. Then the obtained NPs were evaluated and its in vitro drug release characteristics were investigated. RESULTS: When using PEG4000-PCL2000, PEG4000-PCL1250, PEG2000-PCL2000, PEG2000-PCL1250 as the carrier material to prepare NPs, the average particle size of NPs in turn were 116.1, 110.0, 119.9, 99.1 nm; the zeta potential were -22.4, - 16. 9, -33.5, - 28.8 mV; the entrapment efficiency were 88.29%, 83.10%, 80.67%, 77.46%; and the drug loading were 2.96%, 2.56%, 2.31%, 2.14%, respectively. HCPT-PEG-PCL-NPs all showed a certain degree of sustained-release characteristics and their release mechanisms were fitted to Weibull modle, which showed that the drug release process included passive diffusion and matrix-eroded procedure. CONCLUSION: The HCPT-PEG-PCL-NPs has high entrapment efficiency, drug loading, uniform particle size, and can retard drug release in vitro, so it provides an extensive prospect for clinical application of HCPT.

[Suppressive effect of matrine on cell growth and decreases beta-catenin-dependent transcriptional activity in hepatoma cell line Hep3B].

OBJECTIVE: To investigate the effects of matrine on proliferation, cell cycle, apoptosis rate and beta-catenin-dependent transcriptional activity in cultured hepatoma cell line Hep3B. METHODS: Cell viability was analyzed by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis rate wene determined by flow cytometry analysis, beta-catenin-dependent transcriptional activity was assayed with Dual-Luciferase Reporter System. RESULTS: 72 h of matrine (50 - 800 mg/L) exposure significantly suppresses Hep3B cells growth in a concentration-dependent manner (P<0.01), the 50 percent inhibitory concentration (IC50) was 312.53 mg/L. A higher proportion of apoptotic cells in matrine-treated group than in control groups (21.73 +/- 1.66% vs. 4.39 +/- 1.93%) are shown. Cell proportions in G0/G1 phase were respectively 74.48% and 57.39% in matrine-treated group and control group. Cell proportions in S phase wene respectively 12.94% and 27.67%. Beta-catenin reporter activity was also decreased by matrine treatment in a concentration-dependent manner in Hep3B cells (P<0.05 or P<0.01). CONCLUSION: Through inhibition of beta-catenin-dependent transcription, matrine induces cell cycle arrest and apoptosis, and subsequently suppresses proliferation of hepatoma cells.

[Effects of compound Centella asiatica enema on kidneys coefficient, electrolytes and blood in chronic renal failure rats].

OBJECTIVE: To study the pharmacological effects of compound Centella asiatica enema on chronic renal failure (CRF) rats. METHODS: Rats were divided into control group, CRF model group, Niaoduqing positive control group, compound Centella asiatica enema high, middle and low three groups kidney coefficient, electrolyte levels, hematocrit (HCT), red blood cell counts (RBC), hemoglobin (HGB) content were observed after 30 days's treatment. RESULTS: Compared with the model group, the level of the electrolyte, HCT, RBC, HGB of rats in compound Centella asiatica enema high-doses group and the Niaoduqing group were significantly improved. CONCLUSION: High-doses of compound Centella asiatica enema has significant therapeutic effect on CRF rats.

[Preparation and characterization of hydroxycamptothecin nanoparticles of amphiphilic block copolymer].

OBJECTIVE: To prepare hydroxycamptothecin nanoparticles of amphiphilic block copolymer and study its characterization initially. METHODS: Polyethyleneglycol-polycaprolactone (PEG-PCL) was synthesized and its structure was characterized by 1H-NMR. The HCPT-PEG-PCL-NPs were prepared by solvent-diffusion method using PEG-PCL block copolymer as a matrix and HCPT as an antitumor agent. Then the obtained NPs were evaluated and the physical stabilities of both suspl and freeze drying powder were investigated. RESULTS: The mean particle size of the prepared NPs was 164.5 nm, polydispersity index (PI) was 0.14, drug loading (DL) was 5.49%, entrapment efficiency (EE) was 83.2%, zeta potential was -26.1 mV. The physical stability of freeze drying powder was better and hot environment seemed to be bad for the stability. CONCLUSION: The HCPT-PEG-PCL-NPs increase the solubility of HCPT in water and are valuable for the development of the novel dosage form of HCPT.

[Quantitative determination of 1-deoxynojirimycin in mulberry leaves by high-performance liquid chromatographic-tandem mass/mass spectrometry].

OBJECTIVE: To develop a high-performance liquid chromatographic-tandem mass/mass spectrometric method to determine the concentration of 1-deoxynojirimycin (1-DNJ) in mulberry leaves. METHODS: 1-deoxynojirimycin was separated on an SHIMADZU HRC-NH2 column with the mobile phase consisting of acetonitrile and 0.1% formic acid. The mass spectrometric system equipped with a atmospheric pressure chemical ionization (APCI) interface was operated in Multiple Reaction Monitoring (MRM) mode. RESULTS: The retention time of 1-deoxynojirimycin was 2.87 min, and the calibration curve was linear over a concentration range from 482 microg/L to 2410 microg/L, the average recovery was 95.8%. The detection limit was 53.6 microg/L. CONCLUSION: The method is selective and sensitive for determining 1-deoxynojirimycin in mulberry leaves.

[Thyroid-induced hyperthyroidism Yin-deficiency model of rats].

OBJECTIVE: To establish a rat model of hyperthyroidism Yin-deficiency. METHODS: SD rats separate male and female were randomly divided into four groups, a total of eight groups, each with six. The first group for the control group, 2 to 4 for the experimental drug-treated group, including thyroid-low, medium and high-dose group. In addition to the first group lavage volume of saline, the other groups were given the concentration of 2. 5, 5, 10 mg/kg-saline suspension of the thyroid. Once a day, 30 days for delivery. RESULTS: Compared with the control group, with the exception of Group 4 female rats serum T3, T4, FT3, FT4 values were significantly higher (P < 0.01), serum TSH was significantly lower (P < 0.05), the other treatment groups had different values no significant difference (P > 0.05). Group 4 that female rats modeling was success. CONCLUSION: Administration of thyroid-Suspension 10 mg/ kg for 30 d, female rats hyperthyroidism Yin-deficiency rat model is success.

[Effect of shenshuaining dispersible tablets on the levels of NO, NOS, SOD and MDA in kidney of chronic renal failure rats].

OBJECTIVE: To observe the effect of Shenshuaining dispersible tablets on the levels of NO, NOS, SOD and MDA in kidney of chronic renal failure (CRF) rats induced by adenine. METHODS: Seventy-two Sprague-Dawley rats were randomly divided into 6 groups: normal control group, model control group, Niaoduqing group, Shenshuaining dispersible tablets group(high, middle and low dose). From the second to the sixth group, the rats were prepared for model of CRF by being fed 0.5% adenine. After giving corresponding drugs for 7 weeks, we detected the levels of nitric oxide (NO), total nitric oxide synthase (T-NOS), inducing nitric oxide synthase (iNOS), configuration nitric oxide synthase (cNOS), total superoxide dismutase (SOD), Cu/Zn-SOD and malonaldehyde (MDA) in every rat's kidney tissue. RESULTS: Compared with normal rats, CRF rats' renal tissue's levels of NO, T-NOS, cNOS, T-SOD and Cu/Zn-SOD markedly decreased. Both the iNOS and MDA contents significantly increased. Shenshuaining dispersible tablets could markedly ameliorate above indexes. CONCLUSION: Shenshuaining dispersible tablets can effectively improve CRF rats' antioxidation effect and reduce damnification of free radical and protect the kidney.

A new dimeric stilbene from the lianas of Gnetum parvifolium.

A new dimeric stilbene, gnetifolin P (1), was isolated from the lianas of Gnetum parvifolium, together with seven known compounds 2-8. The structure of compound 1 was determined by extensive NMR and HRESIMS data analyses, and quantum chemical calculations. All the compounds were evaluated for their anti-inflammatory activity. Compounds 1 and 6 inhibited the expression of IL-1ß on LPS induced THP-1 cells with the inhibition rate of 35.78 and 64.67%, respectively, at concentration of 25 µg/mL.

[Preparation of 10-hydroxycamptothecin semisolid lipid nanoparticles and investigation of its stability].

OBJECTIVE: To prepare 10-hydroxycamptothecin-semisolid lipid nanoparticles (HCPT-SSLN) and investigate its stability. METHODS: HCPT-SSLN was prepared by the method of "emulsion evaporation at a high temperature and solidification at a low temperature"; The morphology was examined by transmission electron microscope; The particle size and xi potential were determined by laser granularity equipment; The physical stability of both suspl and freeze drying powder of HCPT-SSLN were investigated. RESULTS: The mean particle size of the prepared HCPT-SSLN was 130.5 nm, drug loading was 2.51%, entrapment efficiency was 79.19%, xi potential was -33.1 mV; Placed at room temperature and 4 degrees C for 6 months, the appearance, particle size and entrapment efficiency of HCPT-SSLN were all stable. Moreover, the freeze drying powder was more stable than the suspl. CONCLUSION: The HCPT-SSLN has high entrapment efficiency and drug loading, uniform particle size, good stability, which initially indicates that HCPT is fit for being incorporated into SSLN.

[Sodium tanshinone IIA sulfonate prevents postoperative peritoneal adhesions in rats by enhancing the activity of the peritoneal fibrinolytic system].

OBJECTIVE: To evaluate the effect of sodium tanshinone IIA sulfonate (STS) in preventing postoperative peritoneal adhesions in rats and explore the mechanisms. METHODS: Sixty SD rats were randomized into 4 equal groups, including a blank control group, adhesion model group, and high-, moderate-, and low-dose STS-treated groups, and were subjected to injuries of the parietal peritoneum and cecum to induce peritoneal adhesions, followed by intraperitoneal administration of saline and STS at the doses of 20, 10, and 5 mg/kg for 7 consecutive days, respectively. Another 15 untreated rats served as the blank control group. The adhesion scores in each group were recorded after the treatments; the activity of tissue-type plasminogen activator (tPA) in peritoneal lavage fluid was measured, tPA/PAI-1 protein ratio in the peritoneal tissue was determined by ELISA, and the expressions of TGF-ß1 and collagen I were detected by immunohistochemistry. The anastomotic healing model was used to assess the impact of STS on wound healing. RESULTS: Intraperitoneal administration of STS effectively prevented peritoneal adhesion without affecting anastomotic healing in the rats. Compared with the adhesion model group, the STS-treated groups showed increased peritoneal lavage fluid tPA activity and tPA/PAI-1 ratio in the ischemic tissues with lowered TGF-ß1 and collagen I expressions in the ischemic tissues. CONCLUSIONS: Intraperitoneal administration of STS can prevent peritoneal adhesion and enhance local fibrinolysis in rats, and these effects may be mediated by TGF-ß signaling pathway.

Chinese medicine compound Changtong oral liquid on postoperative intestinal adhesions.

AIM: The aim of this study was to observe the effect of a Chinese medicine compound Changtong oral liquid (CT) on tissue plasminogen activity (t-PA), plasminogen activator inhibitor (PAI), TGF-beta1 and hydroxyproline (OHP). METHODS: Two sets of animal experiments were performed in the present study. Forty New Zealand rabbits and 48 Sprague-Dawley (SD) rats were assigned randomly to one of the five groups: sham adhesion, adhesion with saline, adhesion with low dosage of the CT, adhesion with middle dosage of the CT and adhesion with high dosage of the CT. t-PA and PAI activity in plasma, OHP and TGF-beta1 expression in adhesion were investigated. Analysis of variance was used to test differences among groups. RESULTS: CT treatment increased plasma t-PA activity in rabbits but decreased TGF-beta1 activity in rats. The data were expressed from low to high dose respectively as follows: t-PA, 46.1+/-8.6 microkat/L, 59.6+/-10.1 microkat/L, 64.0+/-11.5 microkat/L; TGF-beta1 28+/-7.23%, 31+/-3.05%, 30+/-4.04%. There were significant differences compared with saline-treated animals (t-PA 26.4+/-5.1 microkat/L, TGF-beta1 54+/-5.51%). OHP content in cecum of rabbits from middle and high but not low dose of CT lowered significantly as compared with saline-treated rabbits, 0.3641+/-0.1373, 0.3348+/-0.0321, 0.2757+/-0.0497 mg/g vs 0.4183+/-0.0883 mg/g of protein, P>0.05, P<0.05, P<0.05 respectively. The rabbit plasma PAI activity and OHP content in abdominal wall had no difference in all groups. CONCLUSION: CT treatment significantly enhanced t-PA activity in rabbits, but decreased TGF-beta1 content in rats, OHP content in cecum of rabbits, and failed to affect the activity of PAI and OHP content in abdominal wall in rabbits, compared with saline group. The result suggests that CT could effectively prevent adhesions without interfering wound healing.

[Changtong oral liquid for prevention of postoperative intestinal adhesion: an experimental study].

OBJECTIVE: To observe the effect Changtong oral liquid (CTOL), a traditional Chinese drug preparation, on the prevention of postoperative intestinal adhesion. METHODS: Fifty-four male SD rats were randomly divided into 6 equal groups, namely the normal control, model control, Simo decoction (SMD, another traditional Chinese drug preparation), and CTOL groups, and CTOL group included three subgroups treated at low, moderate and high doses. Except for those in the normal control group, all the other rats were subjected to operation with Ellis' method for establishing intestinal adhesion models. After the operation, the rats in the normal control and model groups received intragastric administration of distilled water (10 ml/kg), SMD group received SMD administration in similar manner at the dose of 10 ml/kg, and the three CTOL groups had CTOL at the doses of 4.3, 8.6 and 17.2 g/kg, respectively. On day 7 after surgery, blood sample were taken from all the rats to determine the white blood cell (WBC) count and fibrinogen (FIB) concentrations, and the intestinal adhesion was graded. RESULTS AND CONCLUSION: CTOL evidently reduced the severity of postoperative adhesion and decreased WBC count and plasma FIB level, suggesting the efficacy of CTOL in inhibiting the formation of postoperative intestinal adhesion.

Effect of Changtong oral liquid on serum cytokine concentrations in rats with postoperative intestinal adhesion.

OBJECTIVE: To observe the effects of Changtong oral liquid (CTOL) on the serum levels of tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta(1), interleukin (IL)-1beta, IL-4, IL-6 and IL-10 in rats with postoperative intestinal adhesions. METHODS: Fifty-four male Sprague-Dawley rats were randomized into 6 equal groups, namely the normal control, model, Simo decoction (SMD) groups and three CTOL groups of low, moderate, and high doses, respectively. Intestinal adhesion was induced in the rats of the groups other than the normal control group. The rats in the normal control and model groups received intragastric administration of distilled water (10 ml/kg), and those in the 4 treatment groups had SMD (10 ml/kg) and CTOL (at 4.3, 8.6 and 17.2 g/kg for low, moderate, and high dose groups, respectively). On day 7 after surgery, blood samples were obtained from the rats for measurement of serum cytokine levels with enzyme-linked immunosorbent assay followed by adhesion grading according to a 5-grade scale. RESULTS: CTOL evidently reduced the severity of postoperative adhesions and decreased the serum levels of the proinflammatory cytokines such as TNF-alpha, IL-1beta, TGF-beta(1) and IL-6. However, it had no significant impact on serum levels of the anti-inflammatory cytokines IL-4 and IL-10 in rats with postoperative adhesion. CONCLUSION: Significant indices for postoperative adhesion assessment are established, which provides the experimental basis for evaluating clinical therapeutic effects of postoperative adhesions as well as for developing new therapeutic drugs.

[Determination of dexamethasone sodium phosphate content in Fuyankang cream by high-performance capillary electrophoresis].

OBJECTIVE: To determine the content of dexamethasone sodium phosphate in Fuyankang cream, a preparation for treatment of various skin conditions. METHODS: Dexamethasone sodium phosphate content in Fuyankang cream was determined using high-performance capillary electrophoresis (HPCE) at the detection wavelength of 254 nm, under the optimal condition achieved with a fused-silica capillary tube (60 cm x 75 microm) and 100 mmol/L sodium tetraborate buffer (pH 9.2) at a constant voltage of 25 kV with a sampling time of 10 s at 25 degrees Celsius;. RESULTS: The calibration curve displayed good linear relationship within dexamethasone sodium phosphate concentration range of 20 to 100 microg/ ml r=0.999 8 . The average recovery of dexamethasone sodium phosphate was 99.75% n=5, RSD=1.03% . CONCLUSION: HPCE is simple, quick and sensitive in determination of dexamethasone sodium phosphate content in Fuyankang cream.

[Prostaglandin E2 promotes hepatocellular carcinoma cell proliferation through EP2 prostanoid receptor].

OBJECTIVE: To investigate the effect of prostaglandin E2 (PGE(2)) on the proliferation of cultured hepatocellular carcinoma cells and explore which subtypes of EP prostanoid receptor mediate the action. METHODS: RT-PCR was used to determine COX-2 and EP receptor mRNA expression levels in human hepatocellular carcinoma cell line Hep3B and human normal hepatocyte line QSG7701. Cell counting kit-8 (CCK-8) assay was employed to investigate the effect of PGE(2), selective EP2 receptor agonist butaprost and EP3/EP4 receptor agonist PGE1 alcohol on the proliferation of the cells. RESULTS: COX-2 mRNA was highly expressed in Hep3B cells but scarcely in QSG7701 cells. Hep3B cells expressed the mRNAs for all the EP receptor subtypes, but EP2 and EP4 receptors were much more strongly expressed than EP1 and EP3 receptors. PGE(2) significantly promoted Hep3B cell proliferation in a time- and dose-dependent manner, and 10 µmol/L PGE(2) increased the cell proliferation by 22.57% (P<0.001) after a 48-h incubation; treatment with 0.1, 1.0, and 10 µmol/L PGE(2) for 72 h resulted in significantly increased cell proliferation by 12.13% (P<0.01), 17.58% (P<0.01) and 33.07% (P<0.001), respectively. EP2 receptor agonist butaprost (20 µmol/L) increased Hep3B cell proliferation by 21.96% (P<0.001), but the EP3/EP4 receptor agonist PGE(1) alcohol (2-20 µmol/L) exhibited no significant mitogenic effect in Hep3B cells, and 200 µmol/L PGE(1) alcohol decreased the cell viability. CONCLUSION: Selective activation of EP2 receptor promotes Hep3B cell proliferation, indicating the predominant role of EP2 receptor in mediating the mitogenic effect of PGE2.

[Effect of Changtong oral liquid on fibroblast proliferation in normal and adhesive rat peritoneal tissues].

OBJECTIVE: To investigate the effect of Changtong oral liquid (CTOL) on the proliferation of cultured fibroblasts derived from normal peritoneum (NFs) and adhesive peritoneum (AFs) of rats. METHODS: Twenty male SD rats were randomized into 4 groups, including a normal serum group and 3 CTOL groups with CTOL treatment at low, medium or high doses. Serum samples were obtained from the abdominal arteries of the rats after oral treatment with CTOL for 7 days. The fibroblasts were isolated from the peritoneum by means of tissue culture, and the passage 3-8 cells were cultured with the sera of the normal control and CTOL groups for 24, 48, 72 and 96 h. MTT assay was used to observe the proliferation of the fibroblasts. RESULTS: The dose of CTOL was inversely correlated to the absorbance but positively to the growth inhibition rates. Compared with the NFs cultured in normal control rat serum, the NFs in serum from CTOL groups showed no obviously changes in the absorbance at 24 and 48 h, but displayed significant reduction at 72 and 96 h (P<0.01). Compared with the AFs in normal rat serum, the AFs in the 3 CTOL groups all showed significantly decreased absorbance at 24, 48, 72 and 96 h (P<0.05). At the same time point, the inhibition rate of AFs in low-dose CTOL group showed no significant difference from that in the normal control group, but CTOL at a medium dose resulted in a significantly higher inhibition rate of AFs at 72 h (P<0.05). High-dose CTOL produced significant differences in the inhibition rates of AFs and NFs (P<0.05). CONCLUSION: CTOL can inhibit the proliferation of AFs and NFs in vitro. AFs appear to be more sensitive to CTOL, which has a dose-dependent inhibitory effect of AF proliferation.

Antitumor effects of polysorbate-80 coated gemcitabine polybutylcyanoacrylate nanoparticles in vitro and its pharmacodynamics in vivo on C6 glioma cells of a brain tumor model.

Polybutylcyanoacrylate (PBCA) nanoparticles coated with polysorbate-80 have been extensively studied for delivery of drugs into the animal models; however, 1% polysorbate-80 coated gemcitabine PBCA nanoparticles (GCTB-PBCA-NPs) remain unknown. In this study, we investigated the inhibitory effects of brain targeted 1% polysorbate-80 coated GCTB-PBCA-NPs on C6 glioma cells in vitro and in vivo. GCTB-PBCA-NPs were prepared by emulsion polymerization and freeze drying. C6 glioma cells treated with 1% polysorbate-80 coated GCTB-PBCA-NPs showed poor growth with less cell density and increased detachment. Cell morphology was also greatly altered with nuclear vacuoles, ruptured cells and dead cells. Meanwhile, by flow cytometry, the numbers of cells treated with 1% polysorbate-80 coated GCTB-PBCA-NPs showed increase in G0/G1 phase and decreased in the S phase (P<0.01) compared with the blank control. CCK-8 assay also showed that GCTB could significantly inhibited cell proliferation in a concentration dependent manner. Finally, various preparations were injected (90 mg preparation per kg body weight) into the brain tumor model, which was produced after inoculating C6 glioma cells into Sprague Dawley (SD) rats for 14 days, it was shown that 1% polysorbate-80 coated GCTB-PBCA-NPs could significantly extend the survival time compared with the saline control (P<0.05). Taken together, 1% polysorbate-80 coated GCTB-PBCA-NPs can effectively inhibit the growth of C6 glioma cells in vitro and enhance antitumor activity on brain tumor in vivo.

[Determination of arsenic content in Changtong oral liquid by inductively coupled plasma- mass spectrometry].

OBJECTIVE: To determine arsenic content in Changtong oral liquid, inductively coupled plasma-mass spectrometry (ICP-MS) was employed, which generated linear calibration curves in the range of 5-25 ng/ml for As (r=0.9998). The average recovery of As was 98.94% (n=5, RSD=2.58%).