Sensitive Tumor Cell Line for Annonaceous Acetogenins and High Therapeutic Efficacy at a Low Dose for Choriocarcinoma Therapy.

Annonaceous acetogenins (ACGs) have attracted much attention because of excellent antitumor activity. However, the lack of selectivity and the accompanying serious toxicity have eventually prevented ACGs from entering clinical application. To decrease the side effects of ACGs, the cytotoxicity of ACGs on 10 types of tumor cell lines was investigated by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) test to identify one that was very sensitive to ACGs. Meanwhile, ACGs nanoparticles (ACGs-NPs) were prepared using poloxamer 188 (P188) as an excipient so as to solve the problem of poor solubility and the in vivo delivery of ACGs. ACG-NPs were 163.9±2.5 nm in diameter, negatively charged, and spherical with a high drug loading content (DLC) of 44.9±1.2%. MTS assays demonstrated that ACGs had strong cytotoxicity against JEG-3, HeLa, SiHa, MCF-7, A375, A2058, A875, U-118MG, LN- 229, and A431 cells, among which JEG-3 cell line was extremely sensitive to ACGs with a 50% inhibitory concentration (IC50) value of 0.26 ng/mL, a very encouraging discovery. ACGs-NPs demonstrated very good dose-dependent antitumor efficacy in a broad range of 45?1200 µg/kg on JEG-3 tumor-bearing mice. At a very low dose (1200 µg/kg), ACGs-NPs achieved a high tumor inhibition rate (TIR) of 77.6% through oral administration, displaying a significant advantage over paclitaxel (PTX) injections that are currently used as first-line anti-choriocarcinoma drugs. In the acute toxicity study, the half lethal dose (LD50) of ACGs-NPs was 135.5 mg/kg, which was over 100 times as of the effective antitumor dose, indicating good safety of ACGs-NPs. ACGs-NPs show promise as a new type of and potent anti-choriocarcinoma drug in the future.

Combined administration on You-Gui Yin and low-dose Raloxifene partially attenuates the bone loss in ovariectomized mice through the proliferation and osteogenic differentiation of bone marrow stromal cells.

BACKGROUND: Osteoporosis is a systemic skeletal disease of fragility fractures due to the loss of mass and deterioration of the microarchitecture of bone. PURPOSE: The aim of the study was to assess the osteogenic effects and the underlying mechanisms of the combined administration of You-Gui Yin (YGY) and Raloxifene hydrochloride (RLX) in ovariectomized (OVX) mice. METHODS: First, a classic animal model was used to mimic postmenopausal osteoporosis through the removal of the ovary of mice. Second, the OVX mice were administered YGY, RLX, and YGY + RLX for 12 weeks. Next, the bone microtomographic histomorphometry and bone mineral density (BMD) were assessed by micro-CT, and the biochemical markers of procollagen type I N-terminal propeptide (P1NP) and beta-isomerized C-telopeptide (ß-CTX) in serum were assessed. Finally, primary bone marrow stromal cells (BMSCs) were isolated from the tibia and cultured to evaluate cell proliferation and osteogenic differentiation. RESULTS: The results showed that BMD on the YGY + RLX group was higher than that on the RLX group (p < 0.05) and did not have a significant difference when compared with the sham group. Notably, the YGY + RLX group had a dramatically increased trabecular number (Tb.N) compared with that of the YGY group (p < 0.05). Moreover, the BV/TV (bone volume/total volume) and Tb.N in the YGY + RLX group were higher than that in the RLX group (p < 0.05), and the Tb.Sp (trabecular separation) was lower than that in the RLX group (p < 0.05). Moreover, the serum level of P1NP from the YGY + RLX group dramatically increased when compared with that from the YGY and RLX groups (YGY group: p < 0.05; RLX groups: p < 0.01). Notably, there was no significant difference between the YGY and YGY + RLX groups. In addition, cell proliferation from the co-administration of YGY and RLX was clearly higher than a single use of YGY and RLX (p < 0.01, respectively). The ALP/BCA (alkaline phosphatase/bicinchoninic acid) in the YGY + RLX group was higher than that in the RLX group (p < 0.01). CONCLUSION: Overall, co-administered YGY and RLX could partially attenuate bone loss and were more effective than individually using either one; this outcome might be associated with the proliferation and osteogenic differentiation of BMSCs.

Blood compatibility of polyamidoamine dendrimers and erythrocyte protection.

Dendrimers are global macromolecules with promising biomedical application. In order to investigate their blood compatibility, different generations of polyamidoamine (PAMAM) dendrimers terminated with different types of surface functional groups were incubated with red blood cells (RBC) for time course of haemolysis assay and RBC morphology observation. Cationic PAMAM dendrimers demonstrated obvious generation, concentration and time dependent haemolysis while anionic and neutral PAMAM dendrimers were much less hemolytic. The surface amino groups and the polymeric structure of cationic PAMAM dendrimers played pivotal roles in haemolysis induction; meanwhile the basic pH of cationic PAMAM solutions was an inneglectable factor. Due to the presence of plasma proteins in abundance, whole blood was found tolerable to haemolysis induced by dendrimers, possibly through complex formation of plasma proteins with cationic PAMAM dendrimers. It was deduced that poly-carboxylic acids may have the similar effect as that of plasma proteins. So by means of pH adjustment to the physical 7.4, reduction of administration dose and the combined use of plasma protein (or other protective additives such as negatively charged molecules), the blood compatibility and even biocompatibility of cationic dendrimers may be enhanced to "safe" level.

Novel triterpenoid acyl esters and alkaloids from Anoectochilus roxburghii.

Two novel sorghumol acyl esters, sorghumol 3-O-Z-p-coumarate and sorghumol 3-O-E-p-coumarate, and a novel alkaloid, anoectochine, were isolated from the whole plants of Anoectochilus roxburghii along with one known triterpenoid, sorghumol. Their structures were established by their detailed spectral studies, including two-dimensional NMR ((1)H-(1)H COSY, HSQC and HMBC).

[Bioactive constituents inhibiting TNF-alpha production in fresh rhizome of Pinellia ternata].

OBJECTIVE: To study the bioactive constituents of the fresh rhizome of Pinellia ternata, and provide the scientific basis for quality control. METHOD: Various chromatographic techniques were used to separate and purify the chemical constituents, and their chemical structures were determined on the basis of physical-chemical properties and spectroscopic analysis. The inhibitory effects of the isolated compounds on tumor necrosis factor (TNF)-alpha production in the peritoneal macrophages of mice stimulated with lipopolysaccharide (LPS) were assayed in vitro by microplate colorimetric method. RESULT: Nine compounds were isolated and identified as (E)-p-coumaryl alcohol (1), 3, 4-dihydroxycinnamyl alcohol (2), ferulic acid (3), lariciresinol (4), erythro-guaiacylglycerol-beta-O-4'-sinapyl ether (5), dehydrodiconiferyl alcohol (6) , isolariciresinol (7) , sachaliside 1 (8) and coniferin (9). The inhibitory effect of compounds 1, 2, 3, 8, and 9 were 24.1% , 57.6% , 40.2% , 82.7% , and 62.0% , respectively, against the TNF-alpha production in the peritoneal macrophages of mice stimulated with LPS at a concentration of 10(-5) mol L(-1) in vitro. CONCLUSION: The compounds 1, 2 and 4-8 from genera Pinellia Ten. and compound 9 from P. ternata were isolated for the first time. The compounds 1, 2, 8 and 9 were phenylpropanoids, and 4-7 were lignanoids. The anti-inflammatory effects of the rhizome of P. ternata might relate at the least to compounds 1, 2, 3, 8 and 9.

[Studies on the chemical constituents from herba anoectochili].

OBJECTIVE: To study the chemical constituents in the whole herb of Anoectochiluts roxburghii (Wall) Lindl. METHOD: The chemical constituents were isolated by various column chromatographic methods and structurally elucidated by IR, NMR and MS evidences. RESULT: Fourteen compounds were obtained and there into ten were identified as beta-sitosterol(2), ferulic acid(3), oleanolic acid(6), lanosterol(7), p-hydroxybenzaldehyde(8), 3-methoxyl-p-hydroxybenzaldehlde(9), daucosterol(10),3',4',7-trimethoxy-3, 5-dihydroxyflavone(11), isorhamnetin-3-O-beta-D-rutinoside(12) and rutin(13). CONCLUSION: Compounds 6,7,9 and 11 from genera Anoeetochilus were isolated for the first time.

[GC-MS analysis of essential oil from anthodiums of Chrysanthemum morifolium processed by microwave-airflow and steam calefaction].

OBJECTIVE: Analysis of the constituents of the essential oil extracted from the flowers of the Chrysanthemum morifolium processed by the microwave-airflow and the calefaction after steam process from the State Chrysanthemum GAP bases in Zhejiang Province and to provide scientific basis for quality control. METHOD: The essential oil was extracted by water-steam distillation and separated by GC capillary column chromatography. The components were quantitatively determined with normalization method, and identified by GC-MS. RESULT: From the microwave-airflow dried sample and the calefactively dried sample after steam process, 119 and 175 components were detected, among them fifty and fifty-five components were identified, which were composed of 67.89% and 63.64% of the total essential oil, respectively. CONCLUSION: The yield of the essential oil extracted from the microwave-airflow dried sample was 0.40%, and that from the calefactively dried sample after steam process was 0. 19%. The former is 2.1 times higher than the latter. The components of the essential oil are similar and there are differences between the content of corresponding compounds in the two samples. The results showed that application of microwave-airflow combined drying technology remained original and essential constituents. The production benefit was improved greatly. The grade and quality of C. morifolium enhanced, and it's market selling price was increased compared to the traditional drying calefaction after steam process.