Nano-components derived from Descurainiae Semen Carbonisatum and its hemostatic mechanism / 药学学报

A novel sort of nano-component was extricated and isolated from Descurainiae Semen Carbonisatum (DSC), and its hemostatic component was considered through pharmacological experiments. A muffle furnace was used to prepare DSC at 250 ℃, 300 ℃ and 350 ℃, and the DSC dialysate at each temperature was obtained by the extraction and separation method. Low-resolution transmission electron microscopy (TEM) and high-resolution transmission electron microscopy (HR-TEM) were utilized to characterize the nano-components. Ultraviolet spectroscopy (UV-Vis), fluorescence spectroscopy (FL) and infrared spectroscopy (FTIR) were utilized to measure its optical characteristics and functional group information. The anti-hemorrhagic effects were evaluated by liver bleeding tests and the related hemostatic mechanisms of the obtained nano-components were further assessed by detecting blood coagulation and PLT quantity to discuss the hemostasis mechanism. The experiments complied with the Animal Ethics Committee of Beijing University of Chinese Medicine. TEM results showed that there was a novel type of nano-component in the DSC dialysate bag, which was named DSC nano-components (DSC-NCs). The experimental results of liver bleeding in mice showed that DSC-NCs prepared at 250 ℃, 300 ℃, and 350 ℃ could reduce the bleeding time of mice liver. Among them, DSC-NCs prepared at 350 °C had the best effect. In addition, DSC-NCs prepared at various temperatures can also reduce the prothrombin time (PT) value, increase the fibrinogen (FIB) value and the platelet (PLT) value to varying degrees. DSC-NCs have a certain hemostatic effect, which may be related to the activation of the exogenous coagulation system, the increase of FIB value and the increase of platelet content. This provides a new research direction for exploring the treatment of bleeding diseases, and provides a new perspective for the potential application of DSC-NCs in the medical field.

The effect of Moutan cortex carbonisata nano-components on the blood-cooling and hemostatic / 药学学报

At present, the research of Moutan cortex carbonisata (MCC) mainly focuses on the changes of chemical composition before and after charcoal production, and there is a lack of material basic research directly related to the efficacy at home and abroad. In this study, Moutan cortex, as a precursor, and was calcined to MCC at high temperature. The Moutan cortex carbonisata nano-components (MCC-NCs) were extracted and separated from MCC to explore its cooling-blood and hemostatic effects. In the experiment, the MCC was calcined at a high temperature in a muffle furnace (350 ℃, 1 h), and then MCC-NCs were extracted for MCC, and characterized by transmission electron microscopy and UV-vis absorption spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. In addition, the study evaluated the blood-cooling and hemostatic effects of MCC-NCs. The results showed that MCC-NCs have a size distribution of 0.80-2.8 nm, a lattice spacing of 0.26 nm. MCC-NCs are mainly composed of C, O and N elements and have abundant surface functional groups such as OH, C=O, C-N and C=C. The fluorescence yield of MCC-NCs was 7.18%. The experiments complied with the Animal Ethics Committee of Beijing University of Chinese Medicine. The result indicated that pretreatment MCC-NCs can significantly (P < 0.05) reduce the high, medium, and low viscosity of whole blood and plasma viscosity, and reduce hematocrit, red blood cell distribution width, hemoglobin and red blood cell level. In addition, MCC-NCs significantly reduced the levels of activated partial thromboplastin time, thrombin time and fibrinogen (P < 0.05). The pathological examination results showed that MCC-NCs can significantly reduce lung tissue damage, reduce bleeding and inflammatory cell infiltration. At the same time, it can also significantly reduce the symptoms of gastric mucosal bleeding. In conclusion, the results indicated that MCC-NCs has significantly the effect of blood cooling and hemostasis, and its hemostatic effect is mainly related to the activation of endogenous coagulation pathway or fibrinogen system, which provided a novel strategy for exploring the material basis of traditional Chinese medicine for hemostasis.

Establishment of Material Benchmark Characteristic Spectrum of Houpu Qiwutang / 中国实验方剂学杂志

Objective:To establish a characteristic spectrum to reflect the efficacy of Houpo Qiwutang. Method:Based on the correlation between the efficacy and the pharmacological action of each herb in prescription，the target substances of characteristic map were screened. The extraction solvent，detection wavelength and gradient of the active ingredients were optimized. Peak assignment was made by comparing individual drugs. Q-TOF was used to infer the molecular formula of each peak in the characteristic atlas，and the reference substance was identified by the reference substance. The reference substance was screened out according to the correlation of main efficacy and medicine. Result:The characteristic spectrum of material standard of Houpu Qiwutang was established. Five of the seven herbal medicines were attributed. Nine characteristic peaks were selected and identified by Q-TOF as glycyrrhizin，including naringin，neohesperidin，ammonium glycyrrhizinate，rhein，honokiol，magnolol. According to the main efficacy of Houpo Qiwutang，neohesperidin was selected as reference substances. According to the separation of characteristic peaks and the retention time，the mark peak of the characteristic spectrum was determined. Conclusion:The characteristic spectrum of the material basis of Houpo Qiwutang was established by selecting the characteristic peaks and controlling the key components. This method not only reflects the situation of all the effective chemical components，but also focuses on the control of the key efficacy，so as to provide a theoretical basis for the subsequent development and quality control of Houpu Qiwu Tang.

Effect of Puerarin on the Pharmacokinetics of Baicalin in Gegen Qinlian Decoction () in Mice / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the pharmacokinetics of puerarin (PUE) in Gegen Qinlian Decoction (, GQD), and the effects of PUE dosage variations on the pharmacokinetics of baicalin (BAL) in mice.</p><p><b>METHODS</b>GQD is composed of the concentrated granules of four Chinese herbs. Three dosages with different levels of PUE, including GQD, GQD co-administered with PUE, and GQD co-administration with two times the amount of PUE, were used to research the pharmacokinetics of PUE and BAL in mice. The indirect competitive enzyme-linked immunosorbent assay (icELISA) methods based on an anti PUE-monoclonal antibody (MAb)and BAL-MAb were employed to determine the concentration of PUE and BAL in mice blood.</p><p><b>RESULTS</b>After the co-administration of GQD with PUE, the area under the curves (AUC) of PUE increased 2.8 times compared with GQD. At the dose of GQD co-administration at two times that of PUE, the (AUC) of PUE was almost equal to that of GQD co-administration of PUE, showing non-linear pharmacokinetics. The (AUC) of BAL showed a good dose-related increase of PUE (r=0.993) in the range from 100 to 300 mg/kg, indicating that PUE dramatically affects the absorption of BAL in mice. There was no significant difference in the other pharmacokinetic parameters, such as the first time of maximum concentration (T), the second T, or the mean residence time.</p><p><b>CONCLUSIONS</b>The icELISA methods were successfully applied to pharmacokinetic studies of PUE and BAL in GQD in mice. The dosage variability of PUE of the main ingredient in GQD affects its own pharmacokinetic characteristics and the absorption characteristics of BAL.</p>

Preparation and identification of Jujuboside A artificial antigen / 中国中药杂志

Immunogenic antigen (jujuboside A-BSA) and coating antigen (jujuboside A-OVA) of jujuboside A were synthesized by sodium periodate oxidation method for the first time. Jujuboside A artificial antigen was confirmed by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). The titer and specificity of the antibody in serum of immunized mice were detected by enzyme-linked immunosorbent assay (ELISA). The corrected relation curve of inhibition rate showed that the antibody against Jujuboside A obtained from immunized mice could bind to jujuboside A and the titer was up to 1∶4 000. The jujuboside A artificial antigen was synthesized, which can be used further to preparation of monoclonal antibody and the pharmacokinetics study of jujuboside A in laboratory animals.

Synthesis, identification of artificial antigen of catalpol and preliminary study of immunogenicity / 中国中药杂志

The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine (TCM) and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF-MS) and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1:8000. The artificial antigen of catalpol was successfully synthesized.

Synthesis of artificial chlorogenic acid antigen and preparation of polyclonal antibodies / 中草药

Objective: To synthesize artificial chlorogenic acid (CHA) antigen, to prepare CHA polyclonal antibody, and to lay the foundation for the preparation of immune chip for allergic component analysis. Methods: CHA-bovine serum albumin (CHA-BSA) and CHA-ovalbumin (CHA-OVA) were synthesized by carbodiimide method. The characterization of the synthesis was examined by UV spectrometry and TLC method. The titer and specificity of the antibody in serum of immunised mice were detected by indirect enzyme-linked immunosorbent assay (I-ELISA) and indirect competitive enzyme-linked immunosorbent assay (I-CELISA), respectively. Results: According to the UV and TLC determination, the CHA was successfully conjugated with BSA. The serum antibody titer was 1:128 000 with the linear range of 15.6-250 ng/mL, R2 = 0.995, and Y = -0.10 lnX+1.158. No cross-reaction was detected with glycyrrhizic acid, parietic acid and so on, which were commonly used in the combination with CHA. Conclusion: The preparation of multi-clonal antibody with good sensitivity and specificity against CHA is successful, which provides a foundation for the trace detection and allergy research of CHA.

Establishment of small molecular monoclonal antibody technology platform in Chinese materia medica / 中草药

Antibody is a tool of vital importance in modern bioscience research, small molecular antibody technology has a broad application prospect in the field of receptor binding analysis, enzyme assays, and quantitative and/or qualitative analytical techniques of Chinese materia medica (CMM) research. In this paper, combining with the research work carried out by our innovation team, we introduced the establishment background of the small molecular monoclonal antibody technology platform in CMM and technical difficulties in antibody preparation. In view of the technology products based on small molecular monoclonal antibody of CMM, such as ELISA test kit, immune affinity chromatography column, colloidal gold test paper, fluorescently labeled antibodies, and antibody microarrays, we explored and practised the various applications based on the small molecular monoclonal antibody technology looking forward to its scientific significances and application values in the field of CMM research.

Synthesis and identification of artificial antigens of paneoniflorin / 中国中药杂志

Oxidation method with sodium iodide was used to synthesize immunogenic antigen (PF-BSA) and coating antigen (PF-OVA) of paeoniflorin. UV spectroscopy showed that paeoniflorin was successfully conjugated with BSA and OVA. After immunized by PF-BSA, the mice can produce anti-paeoniflorin antibodies specifically. The ELISA test results showed the high titer (1:12 800) and specificity (IC50 = 0.791 mg x L(-1)) of the antiserum from mice injected with PF-BSA. Also, the antiserum showed low cross activities against nine traditional Chinese medicine (TCM) of small molecules. These artificial antigens were successfully synthesized and the anti-paeoniflorin antibody well prepared, which provides the experimental basis for the further study of ELISA and its kit.

Synthesis and identification of artificial antigen of forsythin / 中国中药杂志

The establishment of high specificity and sensitivity method of small molecule monoclonal antibody-based immunoassay has a great importance in the study of small molecule compounds in Chinese medicine, wherein synthesis of small molecule artificial antigen is a critical step in the preparation of small molecule antibodies. Oxidation method using sodium iodide was used to synthesize immunogenic antigen (FRn-BSA) and coating antigen (FRn-OVA) of forsythin. UV spectroscopy and thin layer chromatography showed that forsythin was successfully conjugated with BSA and OVA. After immuned FRn-BSA, the mice could specifically produce anti-forsythin antibodies with titer up to 1:8 000, and the linear range was from 1 mg x L(-1) to 100 mg x L(-1). In this paper, the artificial antigen of forsythin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.

New method for analyzing pharmacodynamic material basis of traditional Chinese medicines by using specific knockout technology with monoclonal antibodies / 中国中药杂志

Study on pharmacodynamic material basis of traditional Chinese medicines is one of the key issues for the modernization of traditional Chinese medicine. Having introduced the monoclonal antibody technology into the study on pharmacodynamic material basis of traditional Chinese medicines, the author prepared the immunoaffinity chromatography column by using monoclonal antibodies in active components of traditional Chinese medicines, so as to selectively knock out the component from herbs or traditional Chinese medicine compounds, while preserving all of the other components and keeping their amount and ratio unchanged. A comparative study on pharmacokinetics and pharmacodynamics was made to explicitly reveal the correlation between the component and the main purpose of traditional Chinese medicines and compounds. The analysis on pharmacodynamic material basis of traditional Chinese medicines by using specific knockout technology with monoclonal antibodies is a new method for study pharmacodynamic material basis in line with the characteristics of traditional Chinese medicines. Its results can not only help study material basis from a new perspective, but also help find the modern scientific significance in single herb or among compounds of traditional Chinese medicines.

A new thought of compound compatibility mechanism based on active small molecules monoclonal antibodies in herbs / 中国中西医结合杂志

Compatibility mechanism study of Chinese herbal compound (CHC) has been one of key contents in Chinese medical research, but the present research methods are not suitable for its own features due to its complexity, which has restricted the process of modernization and intemationalization of Chinese medicine. In this paper, we addressed that the compatibility is closely correlated to their in vivo metabolic processes. The preparation of active small molecules monoclonal antibodies in herbs and testing a variety of effective compositions simultaneously using immunoassay can clarify the in vivo metabolism and mutual interactions of Chinese herbs, which is a new thought of studying the compound compatibility mechanisms.

Study on Antihypertensive time-effect and dose-effect of Sancao Jiangya decoction on SHRs / 中华中医药杂志

Objective: To investigate the antihypertensive time-effect and dose-effect features of Sancao jiangya decoction(SCD).Methods: The blood pressure of spontaneously hypertensive rats at different time points were measured after treatment with Sancao jiangya decoction of low,middle,high concentration by tailartery blood pressure measurement for conscious rats.Results: The blood pressure was decreased at 2 hours after drug taken,there were significant dose-effect relationship between the antihypertensive effect and the low,middle,high dose.At 4h after drug taken,the high,middle dose had dose-effect correlation,but the low-dose had no antihypertensive effect.Further research on the middledose shows that the blood pressure reduced at 1h after drug taken,and the stable antihypertensive effect was keeping during 1-4h,the blood pressure began to rise at 6h,and got back to the level before drug taken at 8h.Conclusion: To choose the Middle-dose(10.4g crude drug/kg body weight) and 2h after drug taken is appropriate for SCD's use.This result laid a substantial foundation for further research on effects evaluation and mechanism of antihypertensive medicine.

Study on the Property of TCM Syndrome of Spontaneous Hypertension Rats / 中医杂志

Objective To establish the disease-syndrome integrated animal models suitable for the studies of TCM through differen- tiating the property of TCM syndromes of spontaneous diseased animal models.Methods With the observation on general behaviors, irritable degree,turning endurance time,pain threshold,urine and stools,luster of hair,growing speed of hair,body weight,tongue condition,degree of eyeball protruding,conjunctiva chroma,blood pressure,heart rate,etc.of spontaneous hypertension rats(SHR) and the comparison with normal rats,the study was carried out on the macroscopic description of property of TCM syndrome of SHR (14～18 weeks of age)and their ethology.Results The blood pressure of SHR at the early stage tended to raise with age growing. Compared with the normal group,the heart rate of SHR rats was obviously quicker(P