The DNA extraction method of animal medicine material is difficult and un-unified,which limits the application of molecular identification to identify animal medicines.In this study,based on the DNA extraction theory of SDS,we assessed the effects of three elements including different EDTA concentrations (0.025 mol·L-1,0.25 mol· L-1,and 0.5 mol· L-1) and whether containing NaCl and Triton X-100 in the lysis buffer on the quality of DNA extracted from different kinds of animal medicine.The optimized lysis buffer was used to extract DNA from 121 commercial animal medicines for original and species identification.The results showed that the lysis buffer of 1％ SDS,0.03 mol· L-1 Tris-HCl,0.25 mol· L-1 EDTA and 0.2 mol· L-1 NaCl had the optimum effect on DNA extraction.This lysis buffer can obtain DNA from animal medicine which is difficult to extract,such as Cicadae periostracum.The DNA extractions of 121 commercial animal medicines by optimized lysis buffer can satisfy the experimental requirements for molecular identification.All samples of commercial animal medicines can be accurately identified to the level of species.It was concluded that optimized lysis buffer can be used in the DNA extraction of different kinds of animal medicines except shells,secretions and processed products.This method provides technique support for the molecular identification of animal medicines.

BACKGROUND:Natural colagen is considered to have low immunogenicity and good biocompatibility relatively. OBJECTIVE:To evaluate the immunogenicity of animal-based colagenin vitro. METHODS:Type I colagen was extracted from bovine tendon after immunogenicity removal. The colagen purity was detected by high performance liquid chromatography and residual DNA was measured quantitatively by fluorescence staining. Fifty BALB/c mice were randomly divided into five groups: subcutaneous injection of normal saline solution (negative control), bovine colagen (positive control), 33.4, 66.8, 133.4 mg/kg of bovine tendon colagen, respectively, once a day. After 12 days of continuously subcutaneous injection, lymphocyte proliferation, and cel classification and NK cel kiling function of mice were detected; after 3 weeks of continuous injection, the spleen, liver, spleen and lung tissue of mice were taken for histological examination. RESULTS AND CONCLUSION:Compared with the standard type I colagen, the purity of purified type I bovine colagen reached more than 99%, but the residual DNA was below 1 mg/L which was far less than the residue level of conventional cel-free DNA in the matrix (dry weight: 50-100 μg/g). After 12 days of continuous injection, there were no changes in lymphocyte proliferation, NK cel kiling function and the proportion of lymphocyte subsets. After 3 weeks of injection, the spleen and lymph sheath of mice around the smal artery became thickened in the 66.8 and 133.6 mg/kg bovine tendon colagen groups, which could cause accidental liver injury and lung injury, but the splenic corpuscle germinal center area had no change. These findings indicate that continuously subcutaneous injection of animal-based colagen can cause the lower lymphocyte immune response to the spleen of BALB/c mice, which may cause accidental liver and lung injuries.

In recent years, with the rapid development of Chinese materia medica (CMM) industry, its clinical applications have become more and more widespread. While, adverse reactions of CMM have also become increasingly prominent. However, for adverse reactions of some CMM, the applications of conventional toxicology studies cannot draw definitive conclusions. These CMM, which were not defined as toxic drugs in traditional Chinese medicine (TCM) theories, have unknown potential toxicities and affect the safety in their clinical use. This paper reviewed recent advances in studies on potential toxicity of non-toxic CMM. It analyzed and summarized potential toxic compounds among them, and introduced application for metabolomics researches on potential toxicities in non-toxic CMM.

Objective To study the potential protective and toxic effects of Acanthopanax senticosus harms(AS)in rats. Methods Twenty male SD rats were divided into the AS-treated and control groups by random number table method. Each group comprised 10 rats. The rats in the AS-treated group were gavage-fed with AS extracts 31. 6 mg/ kg(equivalent to crude drug 0. 386 g/ kg in human)once daily for 20 days and the volume of the drug was 10 ml/ kg. The rats in the control group received an equal volume of saline once daily for 20 days. The urinary samples of 24 h from rats in the 2 groups which were gavage-fed for 1,5,10,15,and 20 days were collected respectively. Ultra-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry was used to detect the ion intensities of metabolites from urinary samples. The ion intensities of urinary metabolites at different time points in the 2 groups were compared. The ion intensity of each metabolite was normalized with respect to the total ion intensities to generate a data matrix. The data matrix at different time points were processed by partial least-squares-discrimination analysis(PLS-DA)of EZinfo software. Characteristic metabolites were screened according to the PLS-DA score plot. Two sets of data matrix with relatively large distance were further processed by orthogonal partial least-squares-discrimination analysis(OPLS-DA)and the variable importance of project(VIP)scores were calculated. Characteristic metabolites induced by AS intervention were screened from variables with VIP > 1 and P < 0. 05 and their mass-charge ratios were detected and compared to the information in Human Metabolome Database(HMDB),and the corresponding metabolites were identified at last. The potential protective or toxic effects of AS in rats represented by these metabolites were analyzed through literature review. Results PLS-DA score plot showed that the urinary metabolic profiles at different time points in the 2 groups had good similarity within groups or time period and presented clustering phenomenon. The urinary metabolic profiles in rats in the 2 groups which were gavage-fed for 1,5,10,15,and 20 days partly overlapped. The urinary metabolic profiles in rats in the AS-treated group with 20 days of AS treatment were far from those in the control group. Two sets of data matrix with relatively large displacement were further processed and 6 potential intervention targets of AS with VIP > 1 and P < 0. 05 were screened and identified as 3-methylguanine,3-methylglutarylcarnitine,3-hydroxydodecanedioic acid, tiglylcarnitine, kynurenic acid,and 13-cis-retinoic acid by comparing with the information of HMDB. At the 20 d of AS-treated group, the expression of 3-methylguanine,kynurenic acid,and 3-hydroxydodecanedioic acid were up-regulated and the expression of tiglylcarnitine,3-hydroxydodecanedioic acid,and 13-cis-retinoic acid were down regulated. By reviewing the related literature,kynurenic acid,13-cis-retinoic acid,and 3-methylguanine may have potential protective effects in rats which were treated with AS and the other 3 metabolites may have potential toxicity. Conclusion The potential intervention effects of AS in rats had two sides,which may have protective effects,and may also have toxic effects.

Objective To study the potential protective and toxic effects of Acanthopanax senticosus harms(AS)in rats. Methods Twenty male SD rats were divided into the AS-treated and control groups by random number table method. Each group comprised 10 rats. The rats in the AS-treated group were gavage-fed with AS extracts 31. 6 mg/ kg(equivalent to crude drug 0. 386 g/ kg in human)once daily for 20 days and the volume of the drug was 10 ml/ kg. The rats in the control group received an equal volume of saline once daily for 20 days. The urinary samples of 24 h from rats in the 2 groups which were gavage-fed for 1,5,10,15,and 20 days were collected respectively. Ultra-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry was used to detect the ion intensities of metabolites from urinary samples. The ion intensities of urinary metabolites at different time points in the 2 groups were compared. The ion intensity of each metabolite was normalized with respect to the total ion intensities to generate a data matrix. The data matrix at different time points were processed by partial least-squares-discrimination analysis(PLS-DA)of EZinfo software. Characteristic metabolites were screened according to the PLS-DA score plot. Two sets of data matrix with relatively large distance were further processed by orthogonal partial least-squares-discrimination analysis(OPLS-DA)and the variable importance of project(VIP)scores were calculated. Characteristic metabolites induced by AS intervention were screened from variables with VIP > 1 and P < 0. 05 and their mass-charge ratios were detected and compared to the information in Human Metabolome Database(HMDB),and the corresponding metabolites were identified at last. The potential protective or toxic effects of AS in rats represented by these metabolites were analyzed through literature review. Results PLS-DA score plot showed that the urinary metabolic profiles at different time points in the 2 groups had good similarity within groups or time period and presented clustering phenomenon. The urinary metabolic profiles in rats in the 2 groups which were gavage-fed for 1,5,10,15,and 20 days partly overlapped. The urinary metabolic profiles in rats in the AS-treated group with 20 days of AS treatment were far from those in the control group. Two sets of data matrix with relatively large displacement were further processed and 6 potential intervention targets of AS with VIP > 1 and P < 0. 05 were screened and identified as 3-methylguanine,3-methylglutarylcarnitine,3-hydroxydodecanedioic acid, tiglylcarnitine, kynurenic acid,and 13-cis-retinoic acid by comparing with the information of HMDB. At the 20 d of AS-treated group, the expression of 3-methylguanine,kynurenic acid,and 3-hydroxydodecanedioic acid were up-regulated and the expression of tiglylcarnitine,3-hydroxydodecanedioic acid,and 13-cis-retinoic acid were down regulated. By reviewing the related literature,kynurenic acid,13-cis-retinoic acid,and 3-methylguanine may have potential protective effects in rats which were treated with AS and the other 3 metabolites may have potential toxicity. Conclusion The potential intervention effects of AS in rats had two sides,which may have protective effects,and may also have toxic effects.

BACKGROUND:Colagen sponges are applied for hemostatic use, wound healing, and residual cavity filing, which have great values in clinical application and scientific research.
OBJECTIVE:To investigate the biological properties, biocompatibility and biodegradability of colagen spongesin vivo.
METHODS: The spatial structure, pore diameter and porosity of colagen sponges were characterized by scanning electron microscopy. Transmission electron microscopy was used to observe the conformation of colagen sponges. The secondary structure and thermal denaturation temperature of colagen sponges were
analyzed by circular dichroism spectrum. Colagen sponges were implanted intramuscularly into the spinal cord of New Zealand rabbits to observe the degradation and absorption and histological changesin vivo.
RESULTS AND CONCLUSION: Colagen sponges had porous structure with varying pore sizes ranging
40-150 μm, the mean pore size of 100 μm, the thickness wal of 1 μm, and a porosity of approximately 95.8%. Colagen sponges had a typical porous structure and periodic light and dark zones. The solution of colagen
sponges had a weak positive band near 220 nm and an intense negative band near 206 nm, which indicated a classic triple helix. And the secondary structure and thermal stability of colagen sponges were similar to that of
liquid colagen. Colagen sponges began to degrade at 4 weeks, and remained 20% at 12 weeks. These sponges had been associated with foreign body response and inflammation within 2 weeks after implantation. With wound healing, inflammatory reactions gradualy reduced and disappeared. During the implantation and degradation of sponges, no significant fibrous capsule formed and no tissue necrosis occurred at implantation site, indicating that colagen sponges have good performance in bioactivity, biocompatibility and degradation.