Cultural Practices for Propagation of Spatholobus suberectus via Cutting into Polybags

In this paper,the propagation of Spatholobus suberectus Dunn.was optimized based on the research results conducted in propagation for many years and the actual need for the planting materials.According to the requirements of Good Agricultural Practices for Chinese Medicinal Drugs,cultural practices for propagation of S.suberectus were summed up and standardized,including the environmental requirements,land preparation,bed preparation,cutting of scions,cuttings treatment,cutting,management of cuttings at the nursery,bagged cuttings ready for transplanting,packing and handling,transportation,and preparation of records for archives.

Stimulating effect of catechin, an active component of Spatholobus suberectus Dunn, on bioactivity of hematopoietic growth factor

Background Hematopoietic growth factor （HGF） is indispensable to hematopoiesis in the body. The proliferation and differentiation of hematopoietic cells must rely on the existence and stimulation of HGF. This study investigated the effect of catechin, an active component extracted from Spatholobus suberectus Dunn （SSD）, on bioactivity of granulocyte-macrophage colony-stimulating activity （GM-CSA）, burst-promoting activity （BPA） and megakaryocyte colony-stimulating activity （MK-CSA） in spleen condition medium （SPCM） of mice to clarify the hematopoietic mechanism of catechin and SSD. Methods Spleen cells of mice were separated and spleen condition medium （SPCM） was prepared from spleen cell culture. Bone marrow cells of mice were separated and cultured in a culture system including 10% （v/v） SPCM （induced by catechin in vivo or ex vivo） for 6 days. Granulocyte-macrophage colony forming units （CFU-GM）, erythrocyte burst-colony-forming units （BFU-E） and megakaryocyte colony-forming units （CFU-Meg） formation were employed to assay the effects of different treatment on the bioactivity of GM-CSA, BPA and MK-CSA in SPCM. Results SPCM induced by 100 mg/L catechin ex vivo could promote the growth of CFU-GM, BFU-E and CFU-Meg, which indicated that catechin could stimulate the production of GM-CSA, BPA and MK-CSA in SPCM. SPCM prepared at the fourth day of spleen cell culture showed the best stimulating activity. The bioactivity of GM-CSA, BPA and MK-CSA in the SPCM prepared after intraperitoneally injecting catechin into mice was also increased. The number of CFU-GM, BFU-E and CFU-Meg gradually increased as the dose of catechin increased and the time of administration prolonged. CFU-GM, BFU-E and CFU-Meg of the high-dose catechin group were significantly higher than those of the control group （P〈0.01） and reached the maximum at the seventh day after administration. Conclusions This study suggests that catechin extracted from the active acetic ether part of Spatholobus suberectus Dunn can regulate hematopoiesis by inducing bioactivity of GM-CSA, BPA and MK-CSA in SPCM of mice. This may be one of the mechanisms for the hematopoietic-supportive effect of catechin and Spatholobus suberectus Dunn.

Antiviral Effects of Aqueous Extract from Spatholobus suberectus Dunn.against Coxsackievirus B3 in Mice

Objective： To investigate the antiviral effects of the aqueous extract of Spatholobus suberectus Dunn. （A.E.）, a Chinese medicinal herb, against coxsackievirus B3 （CVB3）. Methods： The antiviral effects of A.E. against CVB3 in vitro （primarily cultured myocardial cells） and in vivo （BALB/c mice） were determined. Serum pharmacological method was also adopted by in vitro experiments. The effects of A.E. inhibiting the CVB3 mRNA expression were compared by RT-PCR in mice in vivo. Results： A.E. exhibited obvious antiviral effects in vivo, and serum samples obtained from the rats with oral administration of A.E. （10 μg/mL, 5 μg/mL） reduced the virus titers in the infected myocardial cells （3.00±0.70, 3.55±0.52, P〈0.01）. Meanwhile, the viral myocarditis induced by CVB3 was inhibited significantly by A.E., and the 15-day mortality was reduced to 40% and 45% （P〈0.01） in mice treated with A.E. at doses of 50 mg/kg and 100 mg/kg, respectively, while the 30-day mortality was decreased to 45% and 50%, respectively （P〈0.01）. Moreover, the mRNA expression of Coxsackie virus B3 was significantly inhibited by A.E. Conclusion： Aqueous extract of Spatho/obus suberectus Dunn. （A.E.） has inhibitory effect on CVB3 both in vitro and in vivo.

Combined application of extended depth of field imaging, image stitching and polarized microscopy techniques in identification of Spatholobus suberectus

Objective:As traditional techniques for microscopic identification of Chinese medicines currently lack objective and high-quality reference images,here we developed a systemic procedure to be used in microscopic identification of Chinese medicines,which would lead to more objective,effective and accurate identification process.Methods:Spatholobi Caulis(Jixueteng in Chinese)was used as the specimen in the development of such procedure.Jixueteng samples were microscopically examined in bright-and dark-field microscopy.Microscopic images were obtained by regular,EDF,and image stitching techniques.Results:The microscopic images of the characteristics in pulverized Jixueteng were captured,thanks to EDF imaging and image stitching techniques which allowed the detailed and full sighting of each characteristic to be obtained simultaneously.Different layers in anatomical transverse section,including cork,phelloderm,cortex,phloem,cambium,xylem and pith,were distinctively observed.Moreover,by comparing images of bright-and dark-field microscopy,birefringent and non-birefringent components could readily be distinguished.Conclusion:With application of the developed procedure,high-definition,panoramic and microscopic images were acquired,which could be used as the reference images for microscopic identification of Chinese medicines.

Hematopoietic-supportive effect of (2S, 3R)-ent-catechin on marrow-depressed mice

Hematopoiesis is an active process of cell proliferation, differentiation andrelease. It is the process during which hematopoietic stem cells (HSCs) proliferate anddifferentiate to mature blood cells under the effect of hemetopoietic growth factors (HGFs) incertain hematopoietic microenvironment. HSCs are sources of hematopoiesis of a body that canself-renew, differentiate to blood cells of every lineage and maintain the constancy of them. As themajor tissue of hematopoiesis bone marrow is filled with all kinds of blood cells in variousdevelopmental stages. Under the normal conditions, the ordered proliferation and differentiation ofhematopoietic stem cells/hematopoietic progenitor cells ( HSC/HPC) depend on the regulation ofcytokine network. In present, Spatholobus suberectus Dunn (SSD), a traditional Chinese herb medicinethat has been used to invigorate the blood circulation for thousands years, is widely used torebuild blood after chemotherapy and radiotherapy and to treat hematopoietic diseases. Previousreports from our laboratory indicated that (2S,3R)-ent-catechin, epi (2S, 3R )-ent-catechin, gallic( 2S, 3R )-ent-catechin, et al extracted from SSD all could stimulate the proliferation of HPC inmarrow depressed mice, among which (2S, 3R)-ent-catechin obtained from spatholobus genus for thefirst time and whose content was the maxim showed the best effect. In this study, firstly, theeffect of (2S,3R)-ent-catechin on the quality and quantity of HSCs were assayed by detectingCD_(34)^+ expression. Secondly, we investigated the variation of cytokine (GM-CSF and IL-6)sero-level in mice treated with (2S,3R)- ent-catechin by ELISA. And IL-6 mRNA and GM-CSF mRNAexpressions induced by (2S,3R)-ent-catechin were detected simultaneously by RT-PCR technique inorder to clarify its mechanism.