[Two-dimensional zebrafish model combined with hyphenated chromatographic techniques for evaluation anti-osteoporosis activity of epimendin A and its metabolite baohuoside I].

This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.

[Discussion about research progress and ideas on processing mechanism of traditional Chinese medicine].

Study on the processing mechanism of traditional Chinese medicine is the key to science of processing Chinese materia medica and modernization of traditional Chinese medicine. At present, chemical and pharmacology methods are mainly used to discuss the processing principle of efficiency, attenuated, delayed or cooked with different treatment. So that the processing mechanism of Chinese herbal medicine has made breakthrough progress. With the introduction of modern science and technology, biotransformation, intestinal absorption, pharmacokinetics and metabolomics methods have been gradually applied in traditional Chinese medicine processing mechanism. This article summarizes the achievements in the processing mechanism of traditional Chinese medicine in recent years, analyses and discusses some main problems, and points out to in-depth study on absorption and metabolism, strengthening excipient processing mechanism, paying attention to the integration of multiple disciplines and data statistical analysis. Combined with years of exploration and practice, the project group proposes a new idea "traditional Chinese medicine processing mechanism based on coupled effect of chemical composition transformation and intestinal absorption barrier" , which provides reference for the study of the mechanism of traditional Chinese medicine processing.

[Study on preparation of sagittatoside B with epimedin B converted from cellulase].

To prepare sagittatoside B with epimedin B Hydrolyzed from cellulase. With the conversion ratio as the index, the effects of pH value, temperature, reaction time, dosage of enzyme and concentration of substrates on the conversion ratio were detected. L9 (3(4)) orthogonal design was adopted to optimize the preparation process. Hydrolyzed products were identified by MS, 1H-NMR, and 13C-NMR. The results showed that the optimum reaction conditions for the enzymatic hydrolysis were that the temperature was 50 degrees C, the reaction medium was pH 5.6 acetic acid-sodium acetate buffer solution, the concentration of substrates was 20 g x L(-1), the mass ratio between enzyme and substrate was 3: 5, and the relative molecular mass of the reaction product was 646.23. NMR data proved that the product was sagittatoside B. The process is simple and reliable under mild reaction conditions, thus suitable for industrial production.

[Effect of suet oil on formation of self-assembled micelles of flavonoids in Epimedium].

In this paper, the action of suet oil in the preparation of self-assembled micelles of the active flavonoids in Epimedium in the simulated human environment was researched. Twelve suet oil samples were collected from different growing areas and different positions of sheep or goat to simulate the formation of micelles. Then the effects of the fatty acids in suet oil on the preparation of self-assembled micelles were studied furthermore. The results showed that the micelles had a dispersed state and spherical smooth surface. To compare the diameter, potential, encapsulation efficiency and drug loading of the 12 batches micelles, the micelles prepared by the suet oil from Qinghai were more stable and had a higher encapsulation efficiency. The fatty acids in suet oil could promote the formation of self-assembled micelles, but the whole suet oil had a better effect. Above all the study, we confirmed that the suet oil promoted the formation of self-assembled micelles of the flavonoids in Epimedium, it laid foundation for further research about increasing the efficacy of Epimedium and improved the absorption of the active flavonoids in Epimedium.

[Discussion on research ideas of synergistic mechanism of Epimedium fried with suet oil based on self-assembled micelles formation in vivo].

Accessories processing is a major feature of traditional pharmaceutical technology in China How to use modem science and technology to reveal the scientific connotation of accessories processing, is the core of traditional Chinese medicine processing research. As traditional beneficial Chinese medicine, Epimedium can enhance the effect of warming kidney yang after processing with suet oil However, the current literature research from two aspects of chemical constituents and pharmacological action cannot fully explain the processing mechanism of accessories suet oil. According to the characteristics of fatty acids in suet oil with long chain fatty and surface activity, and based on the theoretical basis of self-assembled drug delivery system, as well as the experimental basis of Epimedium flavonoids self-assembled micelles simulated formation and promoting absorption, this paper puts forward the research hypothesis on synergistic mechanism of Epimedium fried with suet oil based on self-assembled micelles formation in vivo. This paper also describes in detail the research ideas and methods of self-assembled micelles enhancing absorption and synergistic mechanism of Epimedium flavonoids, which has the important theory significance and application value to reveal the scientific connotation of Chinese medicine fried with oil.

[Processing mechanism of Epimedium fried with suet oil based on absorption and metabolism of flavonoids].

As beneficial traditional Chinese medicine, Epimedium fried with suet oil can enhance the effect of warming kidney yang. Previous literature studies about processing of Epimedium mainly focused on changes in chemical composition and pharmacological effect. From the angle of flavonoids absorption and metabolism, our group innovatively study the processing mechanism of Epimedium based on active component transformation combined with intestinal absorption barrier. The processing effect of fried Epimedium can be divided into two key aspects of " heat" during processing and processing accessories "suet oil". Through continuous three National Natural Science Foundation items, the research group has clarified the scientific connotation of "heat" during processing with ADME, and explains the synergistic mechanism of processing accessories "suet oil" based on self-assembled micelles formation in vivo for the first time. This paper summarizes the research ideas and results of Epimedium processing mechanism of the project team for many years, and discusses the future research direction and content, in order to provide new ideas and new methods for modern Chinese medicine processing mechanism.

[Construction of research system for processing mechanism of traditional Chinese medicine based on chemical composition transformation combined with intestinal absorption barrier].

Based on practice of Epimedium processing mechanism for many years and integrated multidisciplinary theory and technology, this paper initially constructs the research system for processing mechanism of traditional Chinese medicine based on chemical composition transformation combined with intestinal absorption barrier, which to form an innovative research mode of the " chemical composition changes-biological transformation-metabolism in vitro and in vivo-intestinal absorption-pharmacokinetic combined pharmacodynamic-pharmacodynamic mechanism". Combined with specific examples of Epimedium and other Chinese herbal medicine processing mechanism, this paper also discusses the academic thoughts, research methods and key technologies of this research system, which will be conducive to systematically reveal the modem scientific connotation of traditional Chinese medicine processing, and enrich the theory of Chinese herbal medicine processing.

[Simultaneous content determination of notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and dracorhodin in ZJHX rubber paste by double wavelength HPLC].

Notoginsenoside R1, ginsenoside Rg1, Re, Rb, and dracorhodin from ZJHX rubber paste were analyzed simultaneously by HPLC, with acetonitrile-water as the mobile phase for gradient elution at a flow rate of 1.0 mL x min(-1). The column temperature was 35 degrees C and the sample size was 10 microL. The detection wavelength was set at 203 nm for ginsenoside and 440 nm for dracorhodin, respectively. The results showed that all of notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and dracorhodin could be were separated well by baseline, with the linear ranges of 0.251-5.020 microg (R2 = 0.999 8), 0.520-10.400 microg (R2 = 0.999 9), 0.251-5.010 microg (R2 = 0.999 7), 0.505-10.100 microg (R2 = 0.999 8) and 0.160-3.270 microg (R2 = 0.999 9), respectively. Each component showed a good linear relationship, with the average recoveries ranging from 99.39% to 100.5%. The established method was so simple, accurate and highly reproducible that it could be used for quality control of ZJHX rubber paste.

Oxidative damage to DNA and its relationship with diabetic complications.

OBJECTIVE: To detect the oxidative DNA damage in diabetic patients and to investigate the relationship of oxidative DNA damage with diabetes and diabetic nephropathy. METHODS: Single cell gel electrophoresis (SCGE) was used to detect the DNA strand breaks in peripheral blood lymphocytes, and oxidative DNA damage product and serum 8-OHdG were determined by a competitive ELISA in 47 cases, including 25 patients without diabetic complications, 22 patients with diabetic nephropathy and 25 normal control subjects. RESULTS: Diabetic patients showed greater oxidative damage to DNA. The percentage of comet cells and the length of DNA migration (comet tail length) of peripheral blood lymphocytes were significantly increased in patients with diabetes, and significantly higher in patients with diabetic nephropathy than in diabetic patients without vascular complications (P < 0.05). There was a significant increase in serum 8-OHdG in diabetic patients compared with normal subjects (P < 0.05). Moreover, serum 8-OHdG was much higher in patients with diabetic nephropathy than in diabetic patients without vascular complications (P < 0.05). CONCLUSION: There is severe oxidative DNA damage in diabetic patients. Enhanced oxidative stress may be associated with diabetes, especially in patients with diabetic nephropathy.

[Investigation of RHD 1227A allele in five pedigrees in Zhejiang Han population].

OBJECTIVE: To analysis the genetic mode of Rh DEL phenotype and RHD 1227A allele in Zhejiang Han population through family investigations. METHODS: Rh DEL phenotypes were identified by a serologic adsorption-elution method. Two polymerase chain reaction-sequence specific prime (PCR-SSP) methods which detectED RHD 1227A allele and Rhesus hybrid box, respectively, and a nucleotide sequencing method focused on the exon 9 of RHD 1227A allele were employed to determine the zygosity of RHD allele. RESULTS: All five probands with Rh DEL phenotype harbored a RHD 1227A allele and had a RHD allele deletion, and they were RHD 1227A/RHd heterozygote. One of the parent members was found to contain a RHD 1227A allele and a normal RHD allele in pedigree 1, 2 and 3, respectively. Thus, they were RHD 1227A/RHD heterozygotes and presented normal D positive phenotype. The son of proband No 1. inherited the RHD 1227A allele and presented a normal D positive phenotype due to a RHD 1227A/RHD heterozygote; The offsprings of proband No. 2, No. 4, and No. 5 did not inherit RHD 1227A allele and presented a normal D positive phenotype. CONCLUSION: RHD 1227A allele is an important genetic marker of Rh DEL phenotype; RHD 1227A is recessive to normal RHD allele and dominant to RHd allele; RHD 1227A allele is an ancestral, but not a spontaneously mutated allele.

[Molecular basis of Rh DEL phenotype in Zhejiang Han population].

This study was purposed to investigate the molecular basis of Rh DEL phenotype. Rh DEL phenotypes were identified by a serologic adsorption-elution method, the nucleotide sequences of ten RHD exons and exon-intron boundary regions were evaluated by a RHD gene-specific PCR-SSP (PCR-SSP, polymerase chain reaction-sequence specific primer) and sequencing. The results showed that out of 122 random Rh negative donors 35 Rh DEL phenotypes were identified through serologic method, including 6 RhCCdee (17.14%), 28 RhCcdee (80.00%), and 1RhCcdEe (2.86%). Sequence analysis indicated that all DEL phenotypes harbored a RHD 1227 G > A mutation in exon 9. D zygosity test revealed that 29 DEL phenotypes (28 RhCcdee and 1 RhCcdEe) had one RHD gene deleted, and 6 DEL phenotypes (6 RhCCdee) had homogenous RHD gene. It is concluded that RHD 1227A is an important genetic marker for Rh DEL phenotype in Zhejiang Han population.