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Preparation and in vitro evaluation of ampelopsin-loaded nanomicelles.

Ren-Jie HUANG ; Xue-Li YAN ; Hu-Biao CHEN

China Journal of Chinese Materia Medica.2016;41(6):1054-1058. doi:10.4268/cjcmm20160613

To improve the solubility and antitumor activity of ampelopsin, ampelopsin-loaded nanomicelles from the mixture of pluronic F127 and D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS1000) were prepared by film-thin hydration method, in order to optimize the process conditions and physicochemical properties. The antitumor activities against MCF-7 cells between ampelopsin and nanomicelles were compared by MTT method, respectively. The results showed that the optimal nanomicelles were round with the nanometric size of (22.6±0.5) nm, encapsulation efficiency rate of (80.42±1.13)%, and drug-loading rate of (4.41±0.26)%. The solubility of ampelopsin in mixed nanomicelles significantly increased by 16 times. In different release media, the mixed nanomicelles could release more than 90% of drug in 8 h, and showed stronger cytotoxicity and inhibition against MCF-7 cells (P<0.01). The mixed nanomicelles can be used as new drug delivery system of ampelopsin.

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Matrix formulation of chaizhi cataplasma optimized by D-optimal mixture design combined with multiple mechanical indicators and its in vitro evaluation.

Ye-Wen ZHANG ; Jing-Xin YU ; Jing-Yan WANG ; Qing-Guo RU ; Yan LIU ; Yi-Fei WANG ; Hong-Mei LIN ; Qing WU

China Journal of Chinese Materia Medica.2016;41(6):1046-1053. doi:10.4268/cjcmm20160612

To optimize the matrix formulation of Chaizhi cataplasma (CC) and investigate its release and transdermal absorption properties in vitro. The optimized matrix formulation of cataplasma containing liquid herbal extract is determined by using D-optimal mixture design, with initial bonding strength, endurance bonding strength and gel strength as the evaluating indicators. Modified Franz diffusion cells were used to study the in vitro release and transdermal absorption of geniposide in CC. The optimized matrix formulation of CC contained NP700, aluminum glycinate, tartaric acid, glycerin, PVPK90 and water (9∶0.7∶0.8∶30∶5∶30.5). Cumulative release rate of geniposide in CC was (77.02±3.73)% in 24 h. The percutaneous penetration rate of geniposide was 7.25 μg•cm⁻²•h⁻¹ and the 24 h permeated amount was (156.22±4.90) μg•cm⁻². The optimized CC prepared by the D-optimal mixture design showed a good adhesion and formability. The in vitro release of the geniposide in CC was in accordance with the first order equation, while its in vitro transdermal absorption was close to the zero order equation.

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Chinese medicine preparation supramolecular imprinting templates based on in vitro release characteristics of BYHW sustained-release tablet.

Yan-Tao YANG ; Chun-Ying WU ; Wen-Long LIU ; Ji-Lian SHI ; Jin ZHOU ; Fu-Yuan HE

China Journal of Chinese Materia Medica.2016;41(6):1040-1045. doi:10.4268/cjcmm20160611

The study focused on the in vitro release of Buyanghuanwu (BYHW) elementary osmotic pump sustained release tablets. Its band similarity was calculated by the total quantum statistical moment. Meanwhile, in vitro release characteristics were analyzed to discuss the existence of supramolecular imprinting templates. The results show that the same imprint templates may exist in different structures of traditional Chinese medicine(TCM)'s multi-components. The BYHW sustained release tablets prepared by elementary osmotic pump can meet the objectives of "overall control, synchronous release". However, the supramolecular imprinting templates in TCM compound prescriptions should be further explored, the overall and synchronous release of different components was controlled through imprinting templates, so as to seek the more suitable sustained release preparation technology for multiple components of TCM, and make it in line with the characteristics of TCM.

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Influence of specification on chemical composition of dissolution and hepatocytes toxicity of Polygonum multiflorum.

Yu-Meng LI ; Rui-Yu LI ; Ming NIU ; Chun-Yu LI ; Zhao-Fang BAI ; Wu-Wen FENG ; Cong-En ZHANG ; Peng TAN ; Zhi-Pu HUANG ; Wei-Guang MA ; Jia-Bo WANG ; Xiao-He XIAO

China Journal of Chinese Materia Medica.2016;41(6):1033-1039. doi:10.4268/cjcmm20160610

According to different toxicities of various aqueous extracts of Polygonum multiflorum on hepatocyte, the impacts of chemical composition on the safety of P. multiforum was studied. In this study, 8 main chemical compositions in aqueous extracts of P. multiflorum were determined by the established HPLC method; at the same time, the inhibition ratios of different aqueous extracts of P. multiflorum on L02 cell were determined. Afterwards, the potential compounds related to the toxicity of P. multiforum were tentatively found through a multiple correlation analysis. The results showed that P. multiforum with different chemical compositions exhibited great differences in dissolution. The hepatocyte toxicity of P. multiflorum powder was much greater than P. multiflorum lumps. In addition, three constituents closely related to toxicity of P. multiflorum were found by multiple correlation analysis. The study revealed that chemical composition of P. multiflorum is closely related to the hepatotoxicity, and the hepatotoxicity of P. multiflorum powder is greater than that of other dosage forms. This study indicates that P. multiflorum with different chemical compositions show varying toxicity, which therefore shall be given high attention.

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Which one is more important, raw materials or productive technology?--a case study for quality consistency control of Gegen Qinlian decoction.

Wen ZHONG ; Sha CHEN ; Jun ZHANG ; Yu-Sheng WANG ; An LIU

China Journal of Chinese Materia Medica.2016;41(6):1027-1032. doi:10.4268/cjcmm20160609

To investigate the effect of Chinese medicine raw materials and production technology on quality consistency of Chinese patent medicines with Gegen Qinlian decoction as an example, and establish a suitable method for the quality consistency control of Chinese patent medicines. The results showed that the effect of production technology on the quality consistency was generally not more than 5%, while the effect of raw materials was even more than 30%, indicating that the effect of raw materials was much greater than that of the production technology. In this study, blend technology was used to improve the quality consistency of raw materials. As a result, the difference between the product produced by raw materials and reference groups was less than 5%, thus increasing the quality consistence of finished products. The results showed that under the current circumstances, the main factor affecting the quality consistency of Chinese patent medicines was raw materials, so we shall pay more attention to the quality of Chinese medicine's raw materials. Finally, a blend technology can improve the quality consistency of Chinese patent medicines.

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Cloning and prokaryotic expression analysis of DNA methyltransferas 1(MET1) in Ganoderma lucidum.

Liang-Ping ZHA ; Ya-Jun WANG ; Shuang LIU ; Yu-Yang ZHAO ; Yuan YUAN ; Lu-Qi HUANG

China Journal of Chinese Materia Medica.2016;41(6):1021-1026. doi:10.4268/cjcmm20160608

DNA methyltransferase is the key enzyme in the process of DNA methylation, playing an important role in regulation of gene expression in vivo. According to the Ganoderma lucidum transcriptome data, a full-length cDNA sequence of MET1 from G. lucidum was cloned for the first time, the GenBank registration number is KU239998, and we conducted a comprehensive bioinformatics analysis of the genetic characteristics and spatial structure. The prokaryotic expression analysis showed that E.coli[pET28a(+)-GlMET1] in BL21(DE3) could induce objective protein, shaking the culture at 16 ℃ until the host bacterium(A₆₀₀) was approximately 0.8, and added IPTG to finally concentration of 0.2 mmol•L⁻¹, and then the optimal expression of GlMET1 recombinant protein was accumulated for the induction time of 20 h. The real-time PCR results showed that the expression levels of GlMET1 had obvious differences among varieties of G. lucidum. During the maturity stage, the expression levels of GlMET1 were lower than that in juvenile stage, the results showed that with the growth of G. lucidum, the expression levels of GlMET1 were on the decline. The research provided an important basis for studying the mechanism of DNA methyltransferase thoroughly.

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Mitogen-activated protein kinase genes of Artemisia annua and their expression analysis under Cadmium stress.

Liang-Yun ZHOU ; Chao-Geng LV ; Chuan-Zhi KANG ; Sheng WANG ; Jin-Fu TANG ; Li-Ping KANG ; Lan-Ping GUO

China Journal of Chinese Materia Medica.2016;41(6):1016-1020. doi:10.4268/cjcmm20160607

In order to study Artemisia annua under cadmium stress, whether there are corresponding MAPK genes involved in transduction of the cadmium signal. 17 AaMAPK genes, named AaMAPK1-AaMAPK17 repectively, were finally obtained by using Trinity method for de novo assembly of transcripts from SRA database and BLAST search against AtMAPK genes and determing conserved domain using a series of bioinformatics tools. There exist 16 MAPK genes contained T[D/E]Y conserved domains among the obtained genes. The expressions of these genes were analyzed by Real-time PCR under cadmium stress. The results showed that the expressions level of AaMAPK3 and AaMAPK10 were down-regulated and MAPK7, MAPK9 and MAPK12 were up-regulated. These indicated that there exist corresponding MAPK genes involved in transduction of the cadmium signal.

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Construction of cell factories for production of lupeol in Saccharomyces cerevisiae.

Ting-Ting LIN ; Dong WANG ; Zhu-Bo DAI ; Xue-Li ZHANG ; Lu-Qi HUANG

China Journal of Chinese Materia Medica.2016;41(6):1008-1015. doi:10.4268/cjcmm20160606

Lupane-type triterpenoids, such as betulinic acid, are derived from lupeol and have excellent properties in anti-HIV, anti-cancer activities and so on. For realizing heterogenous production of lupane-type triterpenoids, our research firstly integrated all the seven genes in the MVA pathway in Saccharomyces cerevisiae to increase the supply of squalene (triterpenoids universal precursor) in a single step using the DNA assembler method. Next, cell factories for production of lupeol was constructed by integrating Arabidopsis thaliana lupeol synthetic gene (AtLUP) into chromosome of triterpenoid chassis strain. Results showed that the MVA pathway, about 20 kb nucleotide length, could be assembled in one-pot process and the doubled MVA pathway could significantly improve squalene by 500-fold, reaching 354.00 mg•L⁻¹. NK2-LUP was obtained by introducing AtLUP gene on chromosome, and could produce 8.23 mg•L⁻¹ lupeol. This study supports the possibility of large-scale biosynthetic pathway assembly in S.cerevisiae and lays the foundation of obtaining cell factories for production of lupan-type triterpenoids at the same time.

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Effects of different stocking density and diet on development of Bufo gargarizans tadpoles.

Jun-de LI ; Qun ZHAO ; Yan KANG ; Bo GAO ; Deng-Lin LI ; Yan YANG ; Lu-Qi HUANG

China Journal of Chinese Materia Medica.2016;41(6):1001-1007. doi:10.4268/cjcmm20160605

The effects of four different stocking densities and five different diets on the growth of Bufo gargarizans tadpoles were studied to determine the optimum stocking density and diet. For stocking density experiment, the tadpoles were fed respectively at different density of 200, 500, 1 000 and 2 000 tadpoles per square meter. For diet experiment, the tadpoles were divided into five groups fed respectively with five different diets. The body weight, snout-vent length and tail length were measured every seven days, and mortality was recorded. The results showed that: the survival rates of tadpole before metamorphosis and after metamorphosis were from 68.7% to 96.3% and from 5.7% to 36.0%, respectively; the optimum stocking density is 1 000 tadpoles per square meter for the stocking density had no effect on the survival rate of tadpole before metamorphosis, and the tadpoles had the relative large body weight and survival numbers in 1000 tadpoles per square meter; the diet Ⅱ(37.9% crude protein and 5.7% crude fat), Ⅳ (25.1% crude protein and 4.0% crude fat), and Ⅴ (egg yolk) were all the optimum diets for the diet had no effect on the survival rate of tadpole before metamorphosis and the tadpoles fed with three kinds of diet above had relatively large body weight, and one of these three diets based on their availability and cost should be adopted during breeding period.

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Effects of stocking density and exchanging water frequency on growth and inner quality of Whitmania pigra.

Meng-Meng LI ; Qiao-Sheng GUO ; Hong-Zhuan SHI ; Jia WANG ; Dao-Xin DAI ; Shi-Meng YAN ; Heng-Ji QI ; Xin LU ; Li-Tao LIU

China Journal of Chinese Materia Medica.2016;41(6):995-1000. doi:10.4268/cjcmm20160604

The effects of stocking density and exchanging water frequency on growth, digestive enzyme activity, anti-oxidative enzyme and inner quality of Whitmania pigra Whitman were evaluated with corresponding measures. The results showed that the eventual biomass, specific growth rate, gained weight rate, activities of amylase, lipase, protease, SOD, CAT, and ALP correlated positively with stocking density and negatively with exchanging water frequency (P<0.05). Exchanging water frequency had negative correlation with ammonia nitrogen, nitrite, and hydrogen sulfide while revealed positive correlation with dissolved oxygen in the water. Stocking density and exchanging water frequency showed no significant effects on the contents of moisture, total ash, and acid-insoluble ash. It suggested that the optimum stocking density was 7.5 million per hectare and the appropriate exchanging water interval was 72 h.

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