Development of hybrid-type modified chitosan derivative nanoparticles for the intracellular delivery of midkine-siR NA in hepatocellular carcinoma cells

BACKGROUND： Hepatocellular carcinoma（HCC） is one of the most common cancers worldwide. Most of the patients with HCC lose the surgical opportunity at the time of diagno sis. Some novel therapeutic modalities, like gene therapy, are promising for the treatment of HCC. However, the success of gene therapy depends on two aspects： efficient gene materials and gene delivery vectors. The present study was to develop new chitosan-based nanoparticles for a midkine-si RNA（anti HCC gene drug） delivery.METHODS： The novel gene delivery vector（MixN CH） was syn thesized by hybrid-type modification of chitosan with 2-chloro ethylamine hydrochloride and N, N-dimethyl-2-chloroethylamine hydrochloride. The chemical structure of Mix NCH was char acterized by FT-IR and 1HNMR. The cytotoxicity of Mix NCH was determined by MTS assay. The gene condensation ability and size, zeta potential and morphology of Mix NCH/MK si RNA nanoparticles were measured. The in vitro transfection and gene knockdown efficiency of midkine by Mix NCH/MK si RNA nanoparticles was detected by q RT-PCR and Western blotting. Gene knockdown effect at the molecule level on the proliferation of Hep G2 in vitro was determined by MTS assay.RESULTS： Mix NCH was successfully acquired by aminoalkyl ation modification of chitosan. The Mix NCH could condense MK-siR NA well above the weight ratio of 3. The average size of Mix NCH/MK-si RNA nanoparticles was 100-200 nm, and thesurface charge was about ＋5 m V. Morphologically, Mix NCH/MK-si RNA nanoparticles were in regular spherical shape-with no aggregation. Regarding to the in vitro transfection of nanoparticles, the MixN CH/MK-siR NA nanoparticles reduced MK m RNA level to 14.03%±4.03%, which were comparable to Biotrans（8.94%±3.77%）. Mix NCH/MK-si RNA effectively inhibited the proliferation of Hep G2 in vitro.-CONCLUSION： Mix NCH/MK-si RNA nanoparticles could be effective for the treatment of hepatocellular carcinoma.

Flavonoids from Heartwood of Dalbergia cochinchinensis

Objective To study the flavonoids from the heartwood of Dalbergia cochinchinensis. Methods The chemical constituents were isolated and purified by combination of silica gel, macroporous resin, Sephadex LH-20, and ODS column chromatography. Their structures were identified by means of spectral analysis. Results Fifteen flavonoids were isolated and identified as pinocembrin （1）, liquiritigenin （2）, galangin （3）, 7-hydroxy- 6-methoxyflavone （4）, naringenin （5）, alpinetin （6）, 2,3-dimethoxyxanthone （7）, 6,4＇-dihydroxy-7-methoxy-flavan （8）, mucronulatol （9）, 7,8-dihydroxyflavanone （10）, 5,7,3＇,5＇-tetrahydroxyflavanone （11 ）, 4,2＇,5＇-trihydroxy-4＇-methoxychalcone （12）, isoliquiritigenin （13）, butein （14）, and 3＇,5＇,5,7-tetrahydroxy-6-C-I^-D-glucopyranosyl- flavanone （15）, respectively. Conclusion Compounds 7, 8, ｜ 0, and 15 are isolated from the plants of Dalbergia L. f. for the first time, and compounds 1, 3, 5, 6, 9, 11, 12, and 14 are isolated from this plant for the first time.

Uncarialins J-M from Uncaria rhynchophylla and Their Anti-depression Mechanism in Unpredictable Chronic Mild Stress-Induced Mice via Activating 5-HT_(1A) Receptor

Uncaria rhynchophylla has been widely used to treat central nervous system diseases for a long history.After investigation of U.rhynchophylla,eleven monoterpene indole alkaloids,including four new compounds uncarialins J-M(1-4)and seven known analogues(5-11),were isolated and identified.Their structural characterization was conducted using HRESIMS,1D and 2D NMR,electronic circular dichroism(ECD)spectra,and quantum chemical computations.Compounds 1,2,7,and 9-11 displayed significant ag-onistic effects towards 5-HT_(1A) receptor,and their EC_(50) values were 7.86,732,2.24,1.18,1.52,and 3.75μmol/L,respectively.Furthermore,in vivo experimental results fully revealed that hirsuteine(7)displayed a significant antidepression effect in unpredictable chronic mild stress(UCMS)-induced depression mice mainly via regulating 5-HT_(1A) signaling pathway.Molecular docking and site-directed amino acid mutation verified that amino acid residues Aspll6 and Asn386 were the binding sites of hirsuteine(7)with 5-HT_(1A) receptor.In addition,pre-treatment of mice with WAY 100635 also blocked the anti-depression effect of hirsuteine(7),which further demonstrated that 5-HT_(1A) receptor was a potential target of hirsuteine(7)to effectively treat depression.These findings indicated the therapeutic material basis of U.rhynchophylla and the anti-depression underlying mechanism of hirsuteine(7),and further provided the useful guidance for the development of hirsuteine(7)as a potential antidepressant candidate.