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With matrine(MAT) as the model drug, to prepare nano graphene oxide(NGO)-based MAT in situ gel(MAT-NGO-gel), a kind of drug for tumor treatment in combination with phototheraphy, and investigate the physicochemical properties and anti-tumor effects in vivo of MAT-NGO-gel. First, HPLC method was established to measure the content of MAT in the gel. The ultrasonic method was used to load MAT onto the surface of NGO, and then poloxamer 188 and poloxamer 407 were chosen as the main materials to prepare MAT-NGO-gel. The optimum prescription was selected with the gelation temperature as the index. Finally, the drug loading rate, micromorphology, phototherrmal conversion characteristics and drug release in vitro of MAT-NGO-gel were characterized. In the optimized prescription, the concentration of poloxamer 188 and poloxamer 407 was 2% and 20% respectively, and the mass ratio of NGO and MAT was 1∶1. The gelation temperature and drug loading rate of MAT-NGO-gel prepared by the optimal prescription process was 37.5 ℃ and 16.7%. Under 808 nm laser irradiation, MAT-NGO-gel showed obvious concentration-and time-dependent photothermal conversion characteristics. In vitro release experiments showed that MAT-NGO-gel had temperature-dependent release characteristics. The pharmacodynamics of MAT solution, NGO-gel and MAT-NGO-gel were studied by using S180 tumor-bearing mice and 808 nm laser. The relative tumor volume and body weight of the tumor-bearing mice were plotted over time. After the experiment, the tumor tissues of each group were taken and the histopathological changes were observed by HE staining. The results of pharmacodynamic studies demonstrated that when compared with NS group and NGO-gel group, the body weights of mice in MAT-NGO-gel group and MAT-NGO-gel + laser group were higher, and the relative tumor volume growth was slower. The results of HE stained pathological sections showed that the tumor cells count for the mice in MAT-NGO-gel group and MAT-NGO-gel + laser group was significantly reduced, with obvious nuclear fragmentation and nucleolysis in these two groups. These results suggested that MAT-NGO-gel, especially combined with 808 nm laser, had stronger anti-tumor activity in vivo. The prescription process of MAT-NGO-gel in this experiment was stable and feasible. As compared with MAT solution, MAT-NGO-gel showed obvious sustained and temperature-dependent drug release characteristics. MAT-NGO-gel had much more obvious anti-tumor activity in vivo when combined with 808 nm laser irradiation. This study could provide certain theoretical basis for the therapy of malignant tumor with multiple mechanisms.

In this paper, we analyzed medical records of 40 patients with coronavirus disease 2019(COVID-19), in order to explore the clinical efficacy of Matrine and Sodium Chloride Injection in the treatment of COVID-19. The investigation was based on the results of a previous animal test, which was aimed to investigate and confirme the clinical efficacy of Matrine and Sodium Chloride Injection in the treatment of COVID-19. The animal test demonstrated that Matrine and Sodium Chloride Injection has a significant therapeutic effect on the human coronavirus pneumonia for the model mice. The lung inhibition index reached up to 86.86%. The evaluation was conducted on 40 confirmed cases of COVID-19 treated at Jingzhou Hospital of Infectious Disease(Chest Hospital) of Hubei Pro-vince from January 30~(th) to March 21~(th), 2020. In these cases, patients were treated with other integrated Chinese and Western medicines regimens in the recommended Matrine and Sodium Chloride Injection diagnosis and treatment regimen. The clinical manifestations, laboratory data, nucleic acid clearance time, and imaging data were compared and analyzed before and after treatment. After administration with Matrine and Sodium Chloride Injection, the clinical symptoms of 40 cases were alleviated markedly, and their blood analysis and biochemical indexes returned to normal. The lung CT showed more than 50% of lesion absorption rate, and the viral nucleic acid test showed the average clearance time of patients was 16.6 days, and the average length of hospital stay was 25.9 days. After administration with Matrine and Sodium Chloride Injection, the symptoms of cough and fatigue were alleviated significantly, and the appetite was significantly improved compared with before, especially for patients with gastrointestinal symptoms. Additionally, laboratory indicators, especially absolute value and ratio of lymphocytes and CRP were significantly alleviated. According to the chest CT for short-term review, the absorption of lung lesions was faster than before, especially for grid-like and fibrotic lesions. Compared with antiviral drugs, such as Abidol and Kriging, the nucleic acid clearance time was significantly shorter than the cases treated with Matrine and Sodium Chloride Injection. The clinical effective rate of 40 cases was 100.0%. We believed that Matrine and Sodium Chloride Injection have a good clinical effect in the treatment of COVID-19, and suggested increasing the clinical application and further conducting large-sample-size cli-nical verification.

Through investigation,it was found that the main disease of leaves was grey mold on Dendrobium officinale in Hubei province,which has a great impact on the yield and quality of D. officinale. The identification of morphological and molecular biological was used to prove that the pathogen was Botrytis cinerea. Through test the effect of 5 plant source fungicides and 4 antibiotic fungicides on mycelial growth of strain HS1,which proved 0. 3% eugenol had the best inhibitory effect,EC50 was 0. 29 mg·L

To study the effect of total matrines and extracts from Periplaneta americana on negative endometrial cancer cell JEC of progesterone receptors. After detecting the effect of total matrine, extracts from P. americana and their combination on JEC cells' growth inhibition, cell cycle, P53 and c-erbB-2 gene protein expressions through MTT, flow cytometry instrument and Western blot method, the author found that, (1) MTT: total matrines and extracts from P. americana could inhibit the growth of JEC cell, with significant increase in the inhibitory effect in the combination group. (2) Flow cytometry instrument: the cell cycle at G0/G1 increased after the treatment with total matrines, the cell cycle at G2/M increased after the treatment with extracts from periplaneta americana, and the ratio of G0/G1 cell cycle in the combination group was significantly higher than the other groups, with inhibition in cell growth and statistical difference in inter-group comparison (P0.05). (3) Western blot: the expression level of P53 increased and c-erbB-2 decreased after the treatment with total matrines, extracts from P. americana and their combination on JEC cell, with statistical difference in inter-group comparison (P0.05). The above results suggested that total matrines, extracts from P. americana and their combination could induce cell cycle arrest and inhibit the growth of JEC cell by up-regulating P53 and down-regulating the c-erbB-2 level.

To investigate the antiviral effect of thymopolypeptides combined with 4 kinds of matrine type alkaloids on HepG2.2.15 cells, oxymatrine, sophocarpidine, sophocarpine, and sophoridine (at concentration of 0.2 mmol•L⁻¹ respectively) were respectively combined with thymopolypeptides (0.025, 0.1 g•L⁻¹), and after 48 h and 72 h treatment on HepG2.2.15 cells, the cells and supernatants were collected. The cells activity in various groups was determined by CCK-8 method to evaluate the toxic effects of the drugs on HepG2.2.15 cells. Enzyme linked immunosorbent assay (ELISA) was used to determine HBeAg and HBsAg levels in cellular supernatants. HBV DNA levels in cellular supernatants andcells were quantified with fluorogenic quantitative PCR method; and the expression level of IFN-α in supernatants was detected with CBA method. The results indicated that single thymopolypeptides at 0.025-0.4 g•L⁻¹ had no toxicity to cells. Thymopolypeptides in this concentration range combined with 0.2 mmol•L⁻¹ matrine type alkaloids also had no toxicity to cells. Anti-HBV activity of drug combination was better than that of alkali or thymopolypeptides alone. Thymopolypeptides at 0.025 g•L⁻¹ had better inhibitory effect than thymopolypeptides at 0.1 g•L⁻¹ on intracellular HBV DNA expression, but the inhibitory effect on supernatant HBeAg level was on the contrary. Anti-HBV activity was similar between alkaloids combined with 0.1 g•L⁻¹ and alkaloids combined with 0.025 g•L⁻¹. There was no statistical difference in anti-HBV effect between various combined groups (P

To discuss the effect of matrine on nitric oxide (NO) and asymmetric methylarginine (ADMA) metabolism pathways in serum and tissues of mice with lipopolysaccharide (LPS) -induced intestine tissue inflammation.Kunming mice were randomly divided into five groups: the normal control group, the LPS group and matrine (80, 40, 20 mg x kg(-1) x d(-1)) groups. The mice were intragastrically administered with drugs for 3 d (distilled water of the same volume for the normal control group and the LPS group). One hour after the last intragastrical administration, normal saline or LPS (1 mg x kg(-1)) were intraperitoneally injected. Twelve hours later, serum and tissues were collected to determine NO and ADMA levels and observe the pathological changes of intestinal tissues. The Western blot method was adopted to detect the protein expressions of arginine methyltransferases 1 (PRMT1) and dimethylarginine dimethylaminohydrolase 2 (DDAH2) in intestinal tissues.Compared with the model group, matrine (80, 40, 20 mg x kg(-1) x d(-1)) groups showed lower NO content in serum and tissues, higher ADMA level in serum and increased PRMT1 expression in intestinal tissues, but without effect on DDAH2 expression.Matrine could inhibit LPS-induced intestine tissue inflammation in mice. Its action mechanism is related to the decreased NO content in serum and tissues and increased ADMA level in serum and PRMT1 expression in intestinal tissues.

To prepare matrine double-sensitive colon-specific pellets and study the factors affecting its quality and evaluateing the colon-specific effects of preparation.Matrine enzyme-sensitive pellets core were prepared by carboxymethyl konjac glucomannan as the main carrier material, and coated the core by acrylic resin II and III to prepare matrine double-sensitive colon-specific pellets. The prescription and technology of the matrine colon-specific pellets were studied by the single factor investigation, through the in vitro release test and coating rate determination.The optimized process conditions: FeCl3 concentration is 4.0 g x L(-1), chitosan concentration is 3.0 g x L(-1), carboxymethyl konjac glucomannan concentration is 20 g x L(-1), mixed gel solution pH value is 3. The release of matrine is less than 30% in the simulation of the upper gastrointestinal medium. The release of matrine is close to 100% in simulated full gastrointestinal medium, the coating weight is 7%.The prepared pellets have good colon positioning effect in vitro.

To study the effect of sophoridine against bone cancer pain in bone cancer pain model rats induced by W256 tumor cells and its mechanism.The rat model of bone cancer pain was reproduced by injecting W256 tumor cells into the rat marrow cavity. Ten days after the model establishment, 36 rats were selected and randomly divided into the model control group and the sophoridine treated group. At the same time, other 10 rats with sham-operation were selected to be the normal control group. Since the 15th day after the operation, rats in the treated group had been given sophoridine (25 mg x kg(-1)) for 10 days. The mechanical withdrawal threshold and the thermal withdrawal latency of each group were measured before and after the treatment. After the last treatment, the radiological and histopathological observation shall be conducted for sick legs of all rats. The expressions of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) in tumor tissues were detected by mmunohistochemistry.Sophoridine could significantly increase the mechanical withdrawal threshold and the thermal withdrawal latency (P0.05, P0.01), significantly relief the bone injury caused by W256 tumor cells (P0.05), and notably down-regulate the COX-2 and VEGF expressions in tumor tissues (P0.05).Sophoridine has the effect in relieving pain and inhibiting tumor progression in bone cancer pain rats induced by W256 tumor cells. Its mechanism may be related to the down-regulated expressions of COX-2 and VEGF.

Matrine is one of the main active components extracted from Sophora flavescens, S. subprostrata and S. alopecuroides. In recent years, its anti-tumor activity has attracted wide attention. According to studies, matrine shows the anti-tumor effect through multiple channels such as inducing apoptosis and autophagy of cancer cells, arresting cell cycle, inhibiting tumor cell migration, angiogenesis and NF-kappaB, as well as the synergistic effect with chemotherapeutics. Along with the further studies on matrine's anti-tumor mechanism, it has a broad prospect for development and application in tumor clinical treatment.

A Cleanert Alumina-N-SPE column (0.5 g/6 mL) chromatograpy with 5 mL of chloroform-methanol (7: 3) as eluent, instead of aluminum oxide column (100-200 mesh, 5 g, 1 cm) chromatograpy eluted successively with chloroform and the chloroform-methanol (7:3) (20 mL each), was applied to enrich matrine and oxymatrine in Sophora flavescens. Also, the optimization of the HPLC determination conditions with acetonitrile-ethanol absolute-3% phosphoric acid solution (84: 6: 10) as mobile phase, instead of acetonitrile-ethanol absolute -3% Phosphoric acid solution (80: 10: 10) recorded in Chinese Pharmacopoeia 2010 Edition, was more suitable for determination of matrine and oxymatrine in S. flavescens. This method has advantage of reducing sample handling time and solvent volume and increasing the accuracy and feasibility, which can simplify the procedure for determination of matrine and oxymatrine in S. flavescens.

To establish a new method for quality evaluation and validate its feasibilities by simultaneous quantitative assay of five alkaloids in Sophora flavescens.The new quality evaluation method, quantitative analysis of multi-components by single marker (QAMS), was established and validated with S. flavescens. Five main alkaloids, oxymatrine, sophocarpine, matrine, oxysophocarpine and sophoridine, were selected as analytes to evaluate the quality of rhizome of S. flavescens, and the relative correction factor has good repeatibility. Their contents in 21 batches of samples, collected from different areas, were determined by both external standard method and QAMS. The method was evaluated by comparison of the quantitative results between external standard method and QAMS.No significant differences were found in the quantitative results of five alkaloids in 21 batches of S. flavescens determined by external standard method and QAMS.It is feasible and suitable to evaluate the quality of rhizome of S. flavescens by QAMS.

To study the effect of sophoridine and TLR4/MD-2 blocking agent on pathway of LPS-induced RAW264. 7 macrophage TLR4-NF-kappaB-TNF-alpha in and its pharmacological mechanism of antiendotoxin.RAW264. 7 macrophages were cultured and divided into 5 groups, namely the blank control group, the LPS model group, the sophoridine + LPS group, the TLR4/MD-2 blocking agent + LPS group and the anti-TLR4/MD-2 + sophoridine + LPS group. Cells and cell culture fluids were collected at 120 min after the each group was processed. The expression of TLR4 protein was measured by western blot, the distribution and expression of NF-kappaB protein were measured by immunocytochemistry, and the expression of NF-kappaB and TNF-alpha mRNA were measured by western blot and reverse transcriptase polymerase chain reaction (RT-PCR). The content of TNF-alpha in the cell supernatant was detected by using radioimmunoassay.Compared with the LPS group, the expression of TLR4 protein, NF-kappaB mRNA, the rate of NF-kappaB entry the nucleus, TNF-alpha mRNA and TNF-alpha content in the cell supernatant were significantly decreased in the sophoridine + LPS group (P0.01). The rate of NF-kappaB entry the nucleus and TNF-alpha in the TLR4/MD-2 blocking agent + LPS group and the TLR4/MD-2 blocking agent + sophoridine + LPS group were notablly lower than that of the LPS model group (P0.01), close to that of the blank control group. However, there was no statistical significance between the two groups.TLR4/MD-2 may be one of sophoridine's targets. Sophordine's inhibitory effect on pathway activity of TLR4-NF-kappaB-TNF-alpha may be one of its antiendotoxin mechanisms.

OBJECTIVE To assess the efficacy and safety of compound matrine injection combined with cisplatin chemotherapy for advanced gastric cancer. METHOD It was searched relevant randomized Controlled trials (RCTs) from Cochrane Library, PubMed, EMBASE, CBM, and CNKI etc. The search was finished in February 11, 2010. And it was traced the related references and experts in this field, besides it was also communicated with other authors in order to obtain some certain information that had not been found. RCTs of compound matrine injection combined with cisplatin chemotherapy versus cisplatin chemotherapy for advanced gastric cancer were included. It was evaluated the quality of these included studies and analyzed data by Cochrane Collaboration's RevMan 5.0 software. RESULT Ten RCTs were included meta analysis results suggested that compared with chemotherapy alone, the combination had a statistically significant benefit in healing efficacy (OR = 1.99, 95% CI: 1.26-3.13, P < 0.05) and improving quality of life (OR = 3.83, 95% CI: 2.38-6.15, P < 0.001). Besides, the combination also had a statistically significant benefit in myelosuppression, white blood cell (OR = 0.44, 95% CI: 0.32-0.62, P < 0.001), hematoblast (OR = 0.40, 95% CI: 0.26-0.60, P < 0.001), liver function (OR = 0.33, 95% CI: 0.15-0.75, P < 0.05) and in reducing the gastroenteric reaction (OR = 0.32, 95% CI: 0.16-0.63, P = 0.001), decreasing the of CD3 ( MD = 2.96, 95% CI: 1.724. 20, P < 0.001), CD4 (MD = 9.04, 95% CI: 7.87-10.20, P < 0.001), CD4/CD8 (MD = 0.47, 95% CI: 0.41-0.54, P < 0.001) and NK cells (MD = 5.90, 95% CI: 4.53-7.26, P < 0.001). CONCLUSION Compared with cisplatin chemotherapy, compound matrine injection combined with cisplatin chemotherapy can significantly improve the efficiency, QOL and myelosuppression, and reduce adverse events.

To explore the mechanism of matrine (Mat) induced human erythroleukemia TF-1 cell apoptosis and its effect on SALL4 expression.Different concentrations of the Mat (0.5, 1.0, 1.5, 2.0 g x L(-1) ) were cultured in vitro in TF-1 cells at different time (24, 48, 72 h). Cell proliferation was assayed by MTT. Cell cycle was determined by flow cytometry (FCM). Cell apoptosis was detected by Annexin V and PI double staining method. SALL4 mRNA expression was detected by reverse transcription RT-PCR (RTT-PCR).Administrated with Mat (0.5-2.0 g x L(-1)) after 24, 48, 72 h, the proliferation of TF-1 cells were inhibited (P0.01) , and in dose- and time-dependent manner. Half inhibitory concentration (IC50 ) was 1.0 g L(-1) at 48 h. After 48 h that the Mat acted on TF-1 cells, the proportion of G0/G1 phase cells increased while compared with the control group, and S phase cells decreased (P0.01). Apoptosis were 8.6% , 11.21%, 15.26% , 17.63%, which showed statistically significant difference (P0.01) compared with the control group (5.05%). RT-PCR results showed the ratio between SALL4 mRNA expression and beta-actin (internal reference) expression significantly decreased (P0.01) with Mat dose increased.In a certain range of concentration and time, Mat can inhibit TFT-1 cells proliferation. The mechanism is to make the cells G0/G1 phase blocked, to inhibit SALL4 gene expression and induce cell apoptosis.

OBJECTIVE To study the pharmacokinetics of matrine (MT) intramuscular administration in rat. METHOD Plasma concentration of matrine was determined by HPLC under the following conditions: column (Shim-pack VP-ODS, 4. 6 mm x 150 mm, 5 m); eluent (acetonitrile-0.02 mol ammonium acetate buffer-triethylamine 30: 70: 0.04); flow rate was 1 mL x min(-1) and ultraviolet detection wavelength was set at 220 nm; column temperature 40 degrees C; aliquot injected 20 microL. All data of concentration-time of matrine were treated with pharmacokinetics program DAS 2. 1. 1. RESULT A simple, sensitive and reliable method for determining matrine in rat plasma by HPLC was established. The plasma concentration time profiles of MT fitted in with two-compartment models well, and the main pharmacokinetic parameters found for MT after i. m. infusion were as follows: C(max) = 21.113 9 mg x L(-1), t(max) = 0.75 h, t1/2alpha 1.34 h, t1/2beta = 3.509 h, AUC(0-t) = 90.984 mg x h(-1) x L(-1), AUC(0-infinity) = 100.346 mg x h(-1) x L(-1). CONCLUSION Compare with oral administration, the matrine is absorbed well and distributes fast with intramuscular administration; the absolute bioavailability of matrine is higher. According to this, the pharmacological action is also stronger and duration is longer.

To investigate the epileptic seizure-like effect of Sophora alkaloid sophoridine on electroencepholography (EEG) and its possible characteristic and the mechanism of the seizure-like effect.Chronic electron implantation was employed for the intracranial electroencepholography (IEEG) recording in rat, and the traditional anti-seizure drugs were for the mechanism study in mice.Compared with the medial perforant path (PP) area and the temporal cortex (TC), the granule cells in hippocampus dentate gyrus (DG) area is more sensitive in the kindling effect by sc sophoridine. Under-threshold hypnotic dosage of diazepam and the hypnotic dosage of pentobarbital sodium can block the sophoridine kinded seizure in mice, but the phenytoin sodium can not block the seizure, also the dosage of it can block the maximal electroconvulsive shock (MES) seizure.Sophoridine-induced synchronous oscillations in the hippocampus could elicit the generation and development of seizure. And the hippocampus might play the crucial role and be the original part of the seizure. Sophoridine kinded seizure might belong to clonic seizures, and the diazepam is the ideal agent for the treatment.

To study the anti-cancer effect of matrine (Mat) on U937 cell line and its possible molecular mechanism.The cells were cultured in medium containing either 0.1, 0.2, 0.3, 0.4, or 0.5 g x L(-1) of Mat. The morphological alteration was observed by inverted microscopy and electron microscopy. Cell proliferation was analyzed by Try pan blue staining and MTT. The method of Western Blot was used to detect phosphorylation activity of MAPK.Matrine had a significant inhibitory effect on proliferation of U937 cell line at the concentration of 0.2 g x L(-1). Treated with matrine of 0.2 g x L(-1) for 48 h, U937 cells became smaller and appeared more round than previously. The number of U937 cells showing apoptosis increased with elevation of the concentration of the matrine. Matrine had an ability of inhibiting the activity of ERK and increasing the activities of p38 and JNK to some degree in U937 cells.Matrine can inhibit the proliferation of U937 cell line in vitro and induce its apoptosis possibly through inhibiting the activity of ERK and increasing the activities of p38 and JNK in U937 cells.

To determine the concentration of matrine in rats plasma and tissue and study the pharmacokinetics and tissues distribution of matrine solution (MS), regular matrine liposome (ML) and stealth matrine liposome (LML) after the intravenous administration at a single dose of 15 mg x kg(-1) to rats.Reversed-phase HPLC was used to determine matrine concentration in rats plasma and tissues.The concentration-time curves of MS, ML and LML were fitted to a two-compartment model. The terminal half-life of LML was 2.7-fold higher than MS and 2-fold higher than ML. The area under the plasma concentration curve (AUC) of LML was 63-fold higher than MS and 2.3-fold higher than ML. Tissues distribution results proved that the area under the plasma concentration curve of LML was significantly different from ML and MS (P0.05). The area under the liver and spleen curve of LML was significantly different from ML (P0.05). The ratio between the area under the curve of plasma and the area under the curve of reticulo-endothelial system (Blood/RES) of LML was 5.4-fold higher than ML.Our present studies demonstrate that, compared to MS and ML, LML significantly alters its pharmacokinetics in plasma and tissues targeting.

To prepare colon-targetting tablets of total alkaloids of Sophora alopecuroides and evaluate the effect of pectins of different degree of esterification (DE) on sophoridine release profiles in-vitro.Wet granulation technique was employed to prepare petin-based matrix tablets, then tablets were coated the optimal formulation with Kollicoat MAE 30 DP based on the optimal formulation and analysed their release.Coated formulation E could target total alkaloids of S. alopecuroides to colon and various DE of pectin exerted different effects on sophoridine release. The release of low DE pectin-based matrix tablets coating with Kollicoat MAE 30 DP approximatedly fitted zere-order eqution, which was erosion depended.Low DE pectin-based matrix tablet coating with Kollicoat MAE 30 DP can deliver sophoridine to colon, hence improve the effectiveness of sophoridine.

To study the effect of matrine and oxymatrine on proliferation and the expression of Stat3, Stat5 mRNA in SMMC-7721 cell line.Treated with matrine and oxymatrine, the inhibitory effect on SMMC-7721 cell proliferation was detected by MTT, double fluorescence labeling was applied to measure the apotosis ratios of SMMC-7721cells, the expression of Stat3 and Stat5 mRNA in SMMC-7721 cell line were assessed with RT-PCR.Matrine and oxymatrine could inhibit the proliferation and induce the apoptosis of SMMC-7721 cells and it was time and dose dependent, the expression of Stat3 and Stat5 mRNA in SMMC-7721 cell with matrine and oxymatrine were significantly lower than those in control group (P0.05 or P0.01). Compared with the same dose of matrine and oxymatrine, matrine showed stronger effect on cell proliferation, apoptosis, and Stat3 and Stat5 mRNA (P0.05 or P0.01).Matrine and oxymatrine inhibited the proliferation and induced the of SMMC-7721 cells significantly, the mechanism of which might be related to the down-regulation of stat3 and stat5 mRNA and inhibition of the signaling transduction pathway.

To develop a method for the determination of matrine and oxymatrine in Kushensu injection by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS).The analysis was performed on a Acquity UPLC BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with the gradient elution of 10 mmol x L(-1) amine acetate (pH 8, adjusted by aqueous ammonia) and methanol. The flow rate was set as 0.30 mL x min(-1). The column temperature was maintained at 30 degrees C. The peaks were detected at 210 nm and the injection volume was 2 microL.The calibration curves showed good linearity within the test ranges of 0.084-3.36 microg for matrine and 0.086-3.44 microg for oxymatrine, respectively. The mean recoveries were 103.2%, 98.7%, and the RSD were 1.5%, 1.2%, respectively.The established method is simple, rapid and sensitive, can be used for the quality control of matrine and oxymatrine in Kushensu injection through the manufacturing process and clinical implement.

To investigate the effects of matrine on the anti-tumor efficiency of H22 murine hepatocarcinoma cell-based vaccine modified by TIM2 gene in vivo.The combinant eukaryotic expression vector pIRES2-EGFP-TIM2 was constructed and transfected into H22 cells by lipofectamin. The monoclone of the positive H22-TIM2 cells and negative control H22-EGFP cells were selected by G418 pressure and limited dilution method in turn. The H22 whole-cell-based vaccine were inoculated to establish the tumor-bearing mouse model, and its oncogenicity and immunogenicity were observed in vivo. Then the matrine was administered to the tumor-bearing mice inoculated by H22-TIM2 cells, H22-EGFP cells and H22 cells, and the inhibitory effect of matrine on tumor was studied.The co-expression of EGFP protein and TIM2 mRNA were detected in H22-TIM2 cells. The rate of tumor formation in mice injected of H22-TIM2 cells was 41%, lower than that of H22 cells and H22-EGFP cells injection (92%) in mice. The growth of tumor were significantly inhibited vaccinated with H22-TIM2 cells in mice. The inhibitory rate of tumor (IR) was 69.2% in mice of H22-TIM2 group, higher than that of mice treated with matrine and H22 cells injection, the later was 67.5%. Matrine could dramatically strengthen the anti-tumor efficiency of H22 cells modified by TIM2 gene, with the highest tumor inhibitory rate (IR) (90.6%) in all the experimental mice. The spleen index, populations of CD4-positive lymphocytes and the ratio of CD4-positive to CD8-positive lymphocytes of spleen in mice vaccinated of H22-TIM2 cells were obviously higher than those in the other groups.The oncogenicity of H22 cells is markedly impaired after modified by TIM2 gene. Matrine can strengthen the inhibitory effect of H22-TIM2 cells on tumor in mice. These data give us important clues to further study the biological role of TIM2 gene in tumor immunity and explore the molecular mechanism of matrine in suppressing tumor.

To develop a HPLC method for simultaneous determination of the contents of sophoridine, oxymatrine and oxysophocarpine in seed of Sophora alopecuroides.Chromatographic condition included a Phenomenex Gemini C18 (4.6 mm x 250 mm, 5 microm) column and thmobile phase consisting of a mixture of methanol-0.2% phosphatic acid (7:93). The detection wavelength was at 205 nm. The flow rate was 1.0 mL x min(-1) and column temperature was set at 30 degrees C.All calibration curves showed good linearity (ror = 0.9997) within test ranges. The mean recoveries were 100.0%, 97.0% and 97.0% respectively.This method is simple, rapid and accurate, and suitable for quality control of S. alopecuroides seed.

To investigate the characteristic and influential factors of the degradation of residual pesticides and alkaloids in Radix Sophorae Flavescentis by H2O2.The spiked samples were treated in H2O2 in different reaction time, concentration and pH value. The pesticide residuals were determined by GC-MS, and the contents of alkaloids were determined by HPLC.H2O2 had highly activity in degrading organophosphorus and pyrethroid, but had less activity to organochlorines. The degradation processes of organophosphorus and pyrethroid followed first-order kinetics equations, and were influenced by the pH value, the concentration of H2O2 and reaction time. The contents of alkaloids in Radix Sophorae Flavescentis changed not obviously after treatment with 3 mL x L(-1) H2O2 less than 6 hours under neutral condition.H2O2 is a useful reagent for the degradation of organophosphorus and pyrethroid in crude drug.

To establish a suitable dosage form for a traditional anti-anaphylaxis Chinese medicine of Kushen recipe, and investigate the effect of cutaneous permeation in vitro of the recipe.Techniques of extracting with ethanol and purifying with absorbent resin to obtain alkaloids from Kushen recipe were adopted, while volatile oil was extracted by steam distillation. The extraction was made to gel. The skin from SD rats' abdomen was used as permeability barriers. Then effects of permeation of the aqueous extraction, the purifying extraction and the gel were compared by Valia-Chien and Franz diffusion cell method. HPLC was utilized to quantitate the alkaloids in permeating liquid.In view of the permeation cumulation quantity, the permeation velocity and the lag time of the four kinds of alkaloids, the effect of permeation of purifying extraction was better than the aqueous extraction, and the purifying extraction gel surpassed both the aqueous extraction and the purifying extraction.It was certified that the purifying extraction gel had improved the effect of cutaneous permeation of alkaloids, and it is the befitting dosage form for Kushen recipe to treat anaphylaxis disease in skin.

Taking Chinese compound medicine Yuchangning as a research model, prepared the pH and time dependent Yuchangning tablets for colon-specific delivery (PT-YT-CSD), and evaluated the releasing property in vitro.The coating prescription is filtered by the release extent of matrine and oxymatrine in vitro and the wicking rate of the tablet, which including the category and proportion of film forming agent and porogen, the sort and dosage of fluidizing agent, the increment of weight after coating and so on. The releasing property of the preparation is evaluated by the dissolution tests in vitro through measuring the content of matrine and oxymatrine content.The preparation method of the PYTCSD: After prepared plain tablets, the 95% alcoholic solution of EC and Eudragit II are mixed in a 7:3 EC: Eudragit II ratio and then added in DEP up to 10% of the coating amount, reduced the alcohol concentration to 4% by diluting with ethonal. Tablet was coated by the alcohol solution and the weight of the plain tablet was increased by 3%. The dissolution tests in vitro indicated that matrine and oxymatrine were not dissolved in the simulated gastric juice after 2 h. The accumulative quantities of matrine and oxymatrine were less than 10% in the simulated intestinal juice after 4 h. The quantities of matrine and oxymatrine are 75.7% and 76.8% in the simulated colon juice after 1 h.The PYTCSD was prepared and the preparation could fulfil the aim of delivering in the specific colon in vitro.

To establish a high performance liquid chromatography (HPLC) method for determination of matrine, sophoridine and oxymatrine in Compound Kushen Injection.Alltima amino column was used with acetonitrile-3% phosphoric acid-ethyl alcohol (80:10:10) as the mobile phase and the detection wavelength was at 220 nm.The mean recovery of matrine, sophoridine and oxymatrine in Compound Kushen Injection was 99.51% (RSD 1. 58%), 99.24% (RSD 1.44%) and 100.22% (RSD 1.85%), respectively.The method was simple, rapid, accurate and specific and suitable for quality control of the product.

To study the chemical constituents of the alkaloid fraction of Kushen decoction.Constituents were isolated by different kinds of column chromatography and their structures were elucidated with spectral methods.Eight compounds were isolated and identified as matrine (I), sophoridine (II), sophocarpine (III), sophoramine (IV), oxymatrine (V), oxysophocarpine (VI), aloperine (VII) and sparteine (VIII).All these compounds were isolated from Kushen decoction for the first time. Aloperine was found firstly in Sophora flavescens, or Scutellaria baicalensis, or Rehmannia glutinosa which constituted Kushen decoction.

To investigate the kinetic processes of direct electrooxidation for both matrine and sophoridine in a physiological medium.Their direct electrooxidative behaviors and the parameters of electrochemical kinetics were obtained by cyclic voltammetry (CV), bulk electrolysis with coulometry, chronocoulometry (CC), chronoamperometry (CA).The electrochemical experimental results show that the two totally irreversible oxidation peaks in the region of 0.3-1.2 V vs SCE for both MT and SR were observed, and the electrode reactions processes were diffusion-controlled. Their electron transfer number n and electron transfer coefficient alpha were obtained by CV and electrolysis with coulometry. The diffusion coefficient D and rate constant kf were also calculated by CC and CA.

To observe the base of the interference in correlated biotic active matter obtained multi-drug resistance induced by chemotherapy for different alkaloid, and to supervise the use in clinic to restrain the multi-drug resistant of chemotherapy, and thereby to improve the curative effect.After bestowing subter-dosage unite chemotherapeutant to ascites S180 mouse to set up the mouse models of multi-drug resistance of S180 tumour cell, and giving the mouse matrine, terandrine, oxymatrine and berberine hydrooh loride for 4 weeks, the P170, LRP, TOPOII, Fas and apoposis were determined by flow cytometry.Matrine and terandrine could obviously reduce the express of P170, LRP and the activation of TOPOII, and increase the ratio of the express of Fas and the apoposis of drug resistant tumour cell. And at the same time it could obviously reduce the express of intercellular adhesion molecule(CD54).Matrine and terandrine can interfere in MDR which results from chemotherapeutics by the adjustment of correlated biotic active matter, besides, the different degree of alkaloid effect with different configuration.

To investigate the direct electrochemical behaviors of matrine (MT), oxymatrine (OMT), sophoridine (SR) and oxysophoridine (OSR) at a glassy carbon electrode in a physiological medium, and to determine the contents of commercial MT and SR pharmaceutical products were determined.Their electrochemical behaviors and contents in 0.15 mol x L(-1) NaCl aqueous solutions were obtained by cyclic voltammetry (CV) and square wave voltammetry (SWV) methods.The electrochemical experimental results show that the two totally irreversible oxidation peaks for both MT and SR were observed, which potential values are 0.81 V and 0.92 V, and the electrode reaction processes were diffusion-controlled. But for both OMT and OSR, they have no oxidation peaks observed in the same condition as well. The linear range of the concentration of MT and SR and the contents in their products were obtained by SWV.The research results can be applied in determination of the contents of commercial MT and SR pharmaceutical products.

To establish LC-MS method in the determination of oxymatrine and its metabolite in plasma and investigate their pharmacokinetics in beagle dogs.Lichrospher C18 column (4.6 mm x 250 mm, 5 microm) was used as the analytical column maintained at 25 degrees C. The mobile phase consisted of 10 mmol x L(-1) CH3COONH4 and CH3OH (25:75). Flow rate was 1 mL x min(-1). Electrospray ionization (ESI) was carried out. The ESI ion source was set in positive ion polasity mode. The selective ion monitoring (SIM) was set at m/z 265.1 and 249.2.The linearity ranged from 2 to 5000 ng x mL(-1) (r = 0.9991). The detection of oxymatrine and its metabolite were 0.6 and 0.3 ng x mL(-1). The RSD(%) within day and between day was less than 4.7%. The recovery of this method was more than 96.5%. The disposition was conformed to a two-compartment model. The T(1/2), Tmax, Cmax, MRT, AUC(0--24 h) of oxymatrine were (5.5+/-1.58) h, (1.0+/-0.30) h, (2418.3 +/-970.78) ng x mL(-1), (3.2+/-0.64) h, (5797.4+/-908.16) ng x mL(-1) x h accordingly. The corresponding T(1/2), Tmax, Cmax, MRT, AUC(0--24 h) of matrine were (9.8+/-2.77) h, (1.9+/-1.09) h, (1532.4+/-494.86) ng x mL(-1), (4.4+/-1.97) h, (5530.5+/-1042.65) ng x mL(-1) x h.This assay was highly sensitive, rapid, simple and specific enough for determining concentrations of oxymatrine and its metabolite matrine in plasma of beagle dog.

To study effects of matrine on JM cell strain.Morphologic changes were observed under light microscope with Wright-Giemsa staining, fluorescence microscope with Hoechst 33,258 staining and electron microscope. Alteration of cell cycle of different dose treating groups at the fourth day and 0.8 mg.mL-1 treatment group at the first, second, third, fourth day was analyzed by Flow cytometry. DNA ladder was detected with gel electrophoresis.From the third day after treatment of matrine, typical apoptosis features of cells were observed under light microscope and electron microscope in all test groups, and the features were more prominent with the time prolonging. At fourth day, flow cytometry analysis showed that there were sub-G1 peaks in all groups. From 0.1, 0.2, 0.4, 0.6 to 0.8 g.L-1 treatment groups, the rate of apoptotic cells to total cells were 3.1%, 2. 5%, 13.3%, 40.4%, 48.6%, respectively, and what in the control group was 1.4%; the rate of S phase cells to total cells was 28.9%, 26.1%, 27.7%, 0.9%, 14.2%, what in the control group was 30.4%; the rate of G1 phase cells to total cells was 63. 2%, 67.5%, 68.1%, 75.2%, 83.6%, what in the control group was 41.8%; From the first, second, third to fourth day, the rate of apoptotic cells to total cells of 0.8 mg.mL-1 treatment group were 3.0%, 3.7%, 9.1%, 48.6%, respectively; the rate of S phase cells to total cells was 28.6%, 17.5%, 19.1%, 14.2%; the rate of G1 phase cells to total cells were 45.5%, 77.3%, 77.2%, 83.6%. Gel electrophoresis displayed "DNA ladder" in 0.4, 0.6, 0.8 g.L-1 groups, while 0.1 and 0.2 g.L-1 groups didn't show such result.Matrine can repress DNA synthesis and arrest JM cell strain at G1 phase, sequentially inhibiting the proliferation of the cell. Besides, this alkaloid can induce the apoptosis of JM cells.

The progress in the research on matrine was involved to provide references for the exploitation and utilization of the matrine.Pharmacological functions and mechanism were reviewed according to related experimental studies.Matrine has various pharmacological activities.Matrine has extensive applied prospect and will be developed further.

To establish a method for HPLC fingerprint determination of the alkaloids in S. flavescens.RP-HPLC, linear gradient elution, LC/MS, etc. were used to determine the fingerprint and identify the main peaks in the HPLC fingerprint.A satisfactory method for HPLC fingerprint determination of the alkaloids in S. flavescens. was established, and 5 peaks in the HPLC fingerprint were identified.The perfect fingerprint can be obtained and the method can be used for quality control of S. flavescens.

To study the oxymatrine metabolism induced by human intestinal bacteria and its absorbed metabolites in blood.TLC and HPLC were used to examine oxymatrine and its metabolites, and UV, IR, NMR and MS were used to confirm the chemical structures of the metabolites.Oxymatrine was transformed into matrine by human intestinal bacteria metabolism in vitro. Rats were given orally oxymatrine 100 mg.kg-1 and were decapitated 3 hours after administration, and their blood was taken to examine serum metabolites by TLC and HPLC, which revealed that oxymatrine and matrine were absorbed into blood.Oxymatrine will be transformed into matrine when it is given orally and both of the alkaloids can be absorbed to blood.

To develop a simple and rapid method for the determination of alkaloids in Radix Sophorae Flavescentis (RSF).HPCE buffer: 20 mmol.L-1 phosphates, pH 7.0; V = 20 kV, leff = 50 cm, L = 64.5 cm, i.d. = 50 microns, lambda = 201 nm, T = 25 degrees C, sample size = 10 kPa.s.Matrine(I) and oxymatrine(II) in RSF were completely separated with recoveries of 97.47% and 97.67% respectively.Simple, accurate and reproducible, this method can be used for the quality control of Chinese herbal medicines.

Seven alkaloids were isolated from the seed of Sophora alopecuroides and identified to be oxymatrin, oxysophocarpine, cytisine, matrine, sophocarpine, sophoridine and nicotine respectively by comparing chromactographic and spectral characteristics with authentic known compounds. Nicotine was isolated from Sophora for the first time. The activity of extracts and alkaloids against cancer, virus, dermatophytes and bacteria was carried out in vitro.

Matrine and oxymatrine in Jieeryin and Fuyanshuan were separated and determined by Reversed Phase High Performance Liquid Chromatography, A chromatographic method for the separation and determination of matrine and oxymatrine in these preparations was established. Chromatographic coditions: ODS column with methanol-water-triethylamine(55:45:0.02) as the mobil phase, and UV detection at 215 nm. The method is simple, sensitive and accurate. This study provides the scientific basis for quality evaluation and quality control in making these preparations.

Experiments have been made on the changes of chemical constituents in complex formulas of traditional Chinese medicine. The results show that in the complex formula containing Radix Sophorae Flavescentis, as affected by various conditions of processing such as water, temperature and coexisting reductive substances, the content of oxymatrine tends to decrease gradually to disappear while that of matrine increase gradually.

Six alkaloids have been isolated from the flowers of Sophora viciifolia for the first time. Based on of physico-chemical methods and spectroscopic analysis, their structures have been identified as oxymatrine, oxysophocarpine, sophocarpine, matrine, sophoramine and sophoridine.

The extraction and purificatory crafts for Radix Sophorae Flavescentis were observed through the orthogonal experiment design using matrine as the fixed indicatrix for TLC-scanning. Quantitative analyses of matrine were made on the momentum parameters of the different craft conditions. The results of the three different extraction methods show that the best way is to use 2% gelatin to remove tannin with the least loss of matrine, and that the ethanol extract from water precitation is better than the water extract from ethanol precipitation.