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Objective:To establish an HPLC-DAD-ELSD method for the simultaneous determination of eight main active components in Buyang Huanwu Decoction, including hydroxysafflor yellow A, paeoniflorin, calycosin glycoside, ferulic acid, ononin, calycosin, fermononetin and astragaloside.Methods:Agilent Eclipse XDB-C18 column (250 mm×4.6 mm, 5 μm) was used with acetonitrile-0.1% formic acid as the mobile phase. The flow rate was 1.0 ml/min; the column temperature was 30 ℃; the detection wavelengths were 230 nm (paeoniflorin), 254 nm (calycosin glycoside, ononin, calycosin, fermononetin), 322 nm (ferulic acid) and 403 nm (hydroxysafflor yellow A); the drift tube temperature of the evaporative light scattering detector was 60 ℃; the carrier gas flow rate was 1.6 L/min.Results:Under these conditions, the separation of hydroxysafflor yellow A, paeoniflorin, calycosin glycoside, ferulic acid, ononin, calycosin, fermononetin and astragaloside was good, and the linear relationship was in line with the requirements ( r=0.994 0-0.999 9). The average recovery was 97.8% - 101.4% ( RSD was 1.28% - 3.70%). Conclusion:The method is simple, stable and reproducible, and can be used for the quality control of Buyang Huanwu Decoction.
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Objective:To optimize the ethanol reflux extraction process of total saponins in total saponins of Trillium tschonoskii Rhizome. Methods:On the basis of single factor tests, making the total extraction rate of three main compounds [paris saponin Ⅵ, paris saponin Ⅶ and pennogenin-3- O-α-L-rhamnopyranosyl-(1→4)-[O-α-L-rhamnopyranosyl (1→2)]- O-β-D-glucopyranoside (PRRG)] as the indicator, the optimal extraction parameter was selected with the main influencing factors: the ethanol concentration, solid-liquid ratio, and extraction time by the central composite design-response surface method.Results:The optimal extraction parameter for the ethanol extract of total saponins of Trillium tschonoskii Rhizome was as follows: ethanol concentration 69%, extraction time 1.9 h, and solid liquid ratio 1∶9.7. The binomial fitting complex correlation coefficient r = 0.966 1, and the deviation between the extracted predicted value and the actual value is 4.68%. Conclusion:The central composite design-response surface method is simple and reliable for the optimization of extraction process of total saponins of Trillium tschonoskii Rhizome.
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Objective:Select a suitable macroporous resin for the purification technic of total saponins from Panacis Japonici Rhizoma and determine the parameter of purification technic. Methods:Made the content of total saponins as the index, used static adsorption test and combined the adsorption kinetic parameters to select the type of macroporous resin. By using dynamic adsorption experiment to investigate the technical parameters of the purified macroporous resin extracted from Panacis Japonici Rhizoma. Then the preparation technic of the total saponins of Panacis Japonici Rhizoma was determined. Results:The D101 macroporous resin could absorpt and desorpt total saponins of Panacis Japonici Rhizoma effectively. The optimal purification parameters were as follow: the loading mass concentration was 0.1 g/ml (based on crude drug), and the loading volume was 100 ml (which means the loading volume of resin per ml was equivalent to 3.3 grams of crude drug). During the elution process, distilled water (3 BV) and 20% ethanol (3 BV) were used to remove impurity, and then 70% ethanol elution (6 BV) was used to enrich the total saponins. The flow rate of loading and elution was 0.5 ml/min. The transfer rate of total saponins could reache 85.6%. Conclusion:The D101 macroporous resin can effectively enrich and purify the total saponins of Panacis Japonici Rhizoma, which provides the scientific basis for the development and utilization of Panacis Japonici Rhizoma.
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Family physicians on contract for rural families play an important role in primary medical and health services. This research raises for the first time the practical function and position of rural family physicians on contract. On this basis, a theoretical competency model of rural family physicians was constructed by referring to McClelland′s competency dictionary, relevant policy documents in China and literatures at hand. The model included six level-1 dimension indicators: achievement, management, service, cognition, influence, and self-efficacy, as well as 17 level-2 dimension indicators. At the same time, the paper explained in detail these indicators and compared them with the international indicators of family physicians, highlighting the working characteristics and practical needs of family physicians on contract in rural China.
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Objective To analyze the monosaccharide composition and the relative content of different alcohol precipitations of Panax japonicus polysaccharides.Methods The four components of Panax japonicus polysaccharides were isolated by stepwise ethanol precipitation method.The four components of Panax japonicus polysaccharides were hydrolyzed with trifluoroacetic acid (TFA) and derivated by l-phenyl-3-methyl-5-pyrazolo (PMP),respectively.The monosaccharide composition and relative content were analyzed using LC-FT-ICR-MS and HPLC-UV method.Results The four components of Panax japonicus polysaccharides consisted of glucose,rhamnose,galacturonic acid,galactose,arabinose,and glucose was the main monosaccharide.With the increase of ethanol concentration,the relative content of Ara increased gradually,as while the GalA decreased.Conclusions Precolumn derivation HPLC method was successfully applied for the determination of the monosaccharides in Panax japonicus polysaccharides,and there were differences in the four ethanol precipitations of Panax japonicus polysaccharides.The study can provide a basis for the separation of Panax japonicus polysaccharides.
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The article proposes the construction of network course of Chinese Medicine Preparation Analysis based on Black Board network teaching platform. It states the construction of teaching contents, the implementation of Case-Based Learning, the assistant of practical teaching and the formative evaluation. And it is helpful to improve the quality of classroom teaching, stimulate students self-regulated learning, expand their knowledge, improve their practical ability and innovative consciousness.
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Objective To observe the influence of Xiaoshuan enteric-coated Capsule (XSECC) on neuronal damage in cortex of cerebral hypo-perfusion rats. Methods Rats were divided into a sham group (12), a model group (17), a XSECC large dose group (15), a medium dose group (15) and a low dose group (15) by a random number table. The cerebral hypo-perfusion model was produced by permanent bilateral common carotid artery ligation (2VO). The mixed suspension of XSECC was given orally to rats in the XSECC large dose group (420 mg/kg), the medium dose group (140 mg/kg) and the low dose group (47 mg/kg) once each day for 40 days from the beginning of two hour after ischemia. The expressions of NeuN and Caspase-3 were observed by immunofluorescence staining at 40 days after ischemia. The content of GSH-PX,SOD and MDA were measured by biochemical assay. Results Compared to the model group, the expressions of NeuN (8 716.86 ± 2 539.93, 9 549.31 ± 1 663.26 vs. 7 297.05 ± 1 932.49) were significantly increased in the XSECC large dose group and the medium dose group (P<0.05 or P<0.01);The content of GSH-PX (7.37 ± 1.08 U/mg, 7.77 ± 3.26 U/mg vs. 3.67 ± 2.52 U/mg) was increased in the XSECC large dose group and the medium dose group(P<0.05); The expressions of Caspase-3 (11.65 ± 2.68, 14.05 ± 4.55, 12.60 ± 4.56 vs. 16.80 ± 5.41) were obviously decreased in the XSECC large dose group, the medium dose group and the low dose group, compared with the model group (P<0.05 or P<0.01);The content of MDA (1.44 ± 0.40 nmol/mg, 1.96 ± 1.13 nmol/mg, 2.12 ± 1.19 nmol/mg vs. 3.19 ± 0.98 nmol/mg )was decreased in the XSECC large dose group, the medium dose group and the low dose group when compared with the model group (P<0.05 or P<0.01);The content of SOD (555.61 ± 92.45 U/mg, 607.90 ± 228.45 U/mg, 515.98 ± 184.01 U/mg vs. 348.12 ± 108.84 U/mg) was increased in the XSECC large dose group, the medium dose group and the low dose group when compared with the model group (P<0.05 or P<0.01). Conclusion XSECC could protect injured neurons, which is related to the improvement of free radical metabolism.
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Objective To observe the impact of ginkgo biloba extract in rats with chronic cerebral ischemia on the expression of PSD-95 protein and the content of amino acid neurotransmitter.Methods A total of 42 SD rats were divided into the sham group (n=12), the model group (n=14) and the ginkgo biloba extract group (n=14) by random number table method. Cerebral ischemia rats were produced by permanent ligation of bilateral common carotid arteries . The rats in the ginkgo biloba group were intrgastric administrated with ginkgo biloba extract suspension 28 mg/kg daily for 40 days, since 2 hours later after the surgery. The rats in the sham and model groups were intragastric administrated with equal-Volume nomal saline daily for 40 days, since 2 hours later after the surgery. The expression of PSD-95 protein was detected by immunohistochemistry techniques with image analysis. The content of Glu and GABA in the thalamus was determinated by OPA-pre-column derivatization and HPLC fluorescence detection method.Results The expression of PSD-95 protein (cortex was 212.58 ± 45.02vs.244.20 ± 34.28, thalamus was 132.33 ± 28.32 vs.272.00 ± 62.14) were significantly lower in the cortex and thalamus of the model group than those of the sham group (P<0.01). The content of GABA (6 081.46 ± 2 388.91 mmol/Lvs.8 280.45 ± 3 388.49 mmol/L) in the thalamus of the model group rats was significantly lower than the sham group (P<0.05). Ginkgo biloba extract could significantly improve the expression of PSD-95 protein (cortex was 237.89 ± 34.41 vs.212.58 ± 45.02, thalamus was 226.18 ± 75.80 vs. 132.33 ± 28.32) in the cortex and thalamus of chronic cerebral ischemia rats (P<0.01), and significantly improve the content of Glu and GABA (Glu was 10 523.78 ± 3 639.72 mmol/L vs.6 081.46 ± 2 388.91 mmol/L, and GABA was 18 440.93 ± 7 476.88 mmol/Lvs.11 239.83 ± 4 411.79 mmol/L) in thalamus with chronic cerebral ischemic rats compared with the model group rats (P<0.01).Conclusion Ginkgo biloba extract could regulate the levels of Glu, GABA and selectly regulate the PSD-95 experssion in the cortex and thalamus of cerebral ischemia rats.
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Panax japonicas C.A. Meyd are mostly produced in southwestern China. It is widely used by Tujia and Miao nationality. It has the actions of reinforcing deficiency and being strong, reducing swelling and paln, dissolving stasis and stopping bleeding. Total saponins ofPanax japonicas (TSPJ) are principal active component ofPanax japonicas C.A. Meyd. The researchers found that it had remarkable therapeutic effects on the diseases, especially rheumatism and cardio-cerebrovascular in recent years. This article is to summarize the pharmacological actions of TSPJ and to provide the references for future studies.
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Objectives To investigate the effects of nerve nutrient factors and inhibitory factors on neuronal regen? eration and axonal reconstruction at ischemic neocortical penumbra after focal cerebral ischemia in rats. Methods The rat model of middle cerebral artery ischemia was established using suture-occluded method. The pathological morpholo? gy of brain tissue was examined by using HE staining. The expression levels of GAP-43 SYN, nutrient factor (BDNF VEGF Ang1) and inhibitory factor (NogoA NogoR RhoA) were determined by using Western blotting technique. Results The number of neurons in ischemic penumbra was significantly decreased in model group (P<0.01) than in sham-operat? ed group. The expression levels of GAP-43 were significantly decreased (P<0.05) while the expression levels of NogoA NgR and RhoA in the thalamus were significantly increased (P<0.05) in model group than in sham-operated group. The expression levels of SYN and nutrient factors (BDNF VEGF Ang1) were not different between the model group and sham-operated group. Conclusion The increase in nerve inhibitory factors may contribute to the down-regulation of neu?rogenesis at ischemic neocortical penumbra after focal cerebral ischemia in rats.
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Objective To develop an HPLC method for simultaneous content determination of ginsenoside Rg1, Re and Rb1 in Panacis Japonici Rhizoma.Methods Chromatographic separation was carried out by using an Agilent Poroshell 120 C18 column (4.6 mm × 100 mm, 2.7μm). The mobile phase consisted of acetonitrile-0.1% phosphoric acid with gradient elution at a flowrate of 1.0 mL/min, and the injection volume was 10μL. The detection wavelength and column temperature were 203 nm and 30℃ respectively.Results Ginsenoside Rg1, Re and Rb1 had the baseline separation and were in good linear range. The recovery rates were 99.5%, 103.0% and 100.5% respectively.Conclusion The approach is simple, accurate, with good repeatability and short analysis period, which can determine the contents of ginsenoside Rg1, Re and Rb1 correctly and provide references for quality control of Panacis Japonici Rhizoma.
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This paper discussed the different ways on the pharmacology of traditional Chinese medicine teaching to cultivate students' autonomous learning ability, promoting independent learning through PBL teaching, enhancing students' ability to think independently and practical operation using heuristic teaching.Survey results showed this student-centered, teacher-led, student-teacher interaction style pharmacology teaching model could improve the quality of teaching of Pharmacology.
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This study sought to explore a new compound polyvinyl alcohol-collagen as a wound dressing. To make the polymer, Polyvinyl alcohol (PVA) and collagen type I were put together in the ratio of 3:1, at the same time, polyethlene glycol as porogen was added, and the material was dried by air to be a membrane in shape. Then the ultimate tensile load, the hole diameter, porosity, and water absorption were measured. The cell biocompatibility was tested as well. The results showed the PVA-collagen blend had the average hole 100-150 microm in diameter, and the porosity about 90%. The ultimate tensile load reached 8.10 +/- 0.28 MPa, and water absorption was up to 185.42% +/- 6.93%. 3T3 cells grew well on the PVA-collagen member. Therefore, the PVA-collagen memberane is characterized not only by its ideal biomechanical ability and biocompability, but also by its ideal hole diameter, porosity and water absorption. It may have the potential for use as a wound dressing in vivo.
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Humans , Biocompatible Materials , Chemistry , Collagen , Chemistry , Occlusive Dressings , Polyvinyl Alcohol , ChemistryABSTRACT
BACKGROUND: Polyvinyl alcohol (PVA) displays limitation to cell adsorbability. Can collagen improve the adsorbability of PVA to cells?OBJECTIVE: To develop a novel type composite of PVA and collagen, and explore the feasibility to serve as soft tissue substitute.DESIGN: Single sample observation.SETTING: Guangzhou Red Cross Hospital, Jinan University Medical College, Guangzhou Institute of Traumatic Surgery.MATERIALS: Fifteen New Zealand rabbits of 2.0-3.0 kg, either male or female, were provided by Medical Experimental Animal Center of Guangdong Province. The experiment was carried out in the Laboratory of Guangzhou Institute of Traumatic Surgery, and the experimental procedure was accorded with the animal ethical standards. Bovine typeI collagen was purchased from Guangzhou Trauer Biotechnology Co., Ltd. and PVA-124 from Guangzhou Chemical Reagent and Instrumentation Co., Ltd.METHODS: The experiment was performed in Guangzhou Red Cross Hospital, Jinan University Medical College between July 2003 and December 2006. ①Preparation of PVA-collagen material: 5 g/L bovine type I collagen was mixed with 5% PVA-124 at a ratio of 1 : 1. The mixture was freeze-dried at vacuum until becoming gelatinous. The internal structure was observed under the use of scanning electron microscope. ②Cytotoxicity test: PVA-collagen composite was cut into pieces of 10 mm×5 mm× 1 mm, put into 48-well culture plate after sterilized by Y ray, cultured with 1×104 3T3 cells in each well. Cell growth was observed under scanning electron microscope and laser scanning confocal microscope. ③Embedding test in vivo: Two longitudinal incisions were cut at the two sides of spine. The subcutaneous tissue was separated bluntly to form subcutaneous lacuna. Four pieces of PVA-collagen material were implanted in the lacuna and fixed. Nine specimens and the surrounding tissues were harvested from three rabbits each at one, four, eight and sixteen weeks postoperatively for pathological observation.MAIN OUTCOME MEASURES: The internal structure of gel film under scanning electron microscope, cytotoxicity test and embedding test in vivo results.RESULTS: ①Internal structure of PVA-collagen material:PVA-collagen material showed white gel shape after freeze-drying at vacuum. Penetrating three-dimensional pores were observed in the surface and inner section under scanning electron microscope. ②Cytotoxicity test results showed that 3T3 cells grew normally on the PVA-collagen material. ? Embedding test in vivo results suggested that one week after PVA-collagen implantation, foreign body reaction occurred, and the interface between material and tissue was clear. Four weeks later, only rare lymphocyte infiltration was observed, and a great amount of fibroblast hyperplasia formed collagen fibrils and false simple cuboidal epithelium coating material. In 8 weeks, no lymphocyte infiltration, neutrophilic granulocyte infiltration or foreign body giant cell were found; dense capsule wall and capsule coating material generated from a great amount of fibroblasts were observed. In 16 weeks, extending collagen fibrils were found arranged regularly with shrank nucleus, showing long ovoid or long fusiform in shape; no new formation small vessels, lymphocyte, neutrophilic granulocyte infiltration or foreign body giant cell infiltration were observed. The capsule wall was stable and thinned. CONCLUSION: PVA-collagen composite has good cell compatibility and tissue compatibility but no toxic or adverse effect. It can serve as in vivo implant.
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BACKGROUND: Simple polyvinyl alcohol (PVA) has limited ability to cell adhesion. There are not generally accepted studies on improved effects of collagen protein modified polyvinyl alcohol on cell adhesion and proliferation.OBJECTIVE: To investigate the feasibility of PVA/type Ⅰ college (COL-Ⅰ) as anterior cruciate ligament (ACL) scaffolds in tissue engineering.DESIGN, TIME AND SETTING: The controlled observation experiment was performed at the Fourth Affiliated Hospital, Medical College. Ji'nan University, Guangzhou Red Cross Hospital, Guangzhou Institute of Trauma Surgery from August 2006 to October 2007.MATERIALS: COL-Ⅰ gel was produced by Guangzhou Institute of Trauma Surgery.METHODS: PVA filature was used to weave fascicular scaffolds. NIH-3T3 cell line and human ACL cells were in vitro incubated, amplified, and then implanted on the PVA/COL scaffolds.MAIN OUTCOME MEASURES: The growth of NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds and the secretion of extracellular matrix were observed using scanning electron microscope. Cell compatibility of PVA/COL scaffolds was assessed. Mechanics characteristic of PVA/COL scaffolds was measured by using the electric. tensile force apparatus. Mechanical property of PVA/COL scaffolds was analyzed using the SPSS 11.5 software package.RESULTS: NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds adhered, proliferated, and secreted extracellular matrix. NIH-3T3 cell line highly grew compared with human ACL cells on the PVA/COL scaffolds. The adhered number of NIH-3T3 cell line and human ACL cells was significantly increased on the PVA/COL scaffolds. NIH-3T3 cell line and human ACL cells presented well morphology on the PVA/COL scaffolds. COL-Ⅰ could promote the secretion of extracellular matrix from NIH-3T3 cells, but its effects on human ACL cells were not significant. Tensile force test showed that load-extension curve of the materials was identical to ACL of human and rabbits, and the scaffolds possessed strong flexibility. The maximal load, ultimate stress and elastic modulus were respectively 52.61 N, 14.96 MPa and 202.08 MPa.CONCLUSION: COL-Ⅰ accelerates the adhesion and proliferation of NIH-3T3 cell line and human ACL cells on the surface and in the pore of the PVA/COL scaffolds, promotes the secretion of extracellular matrix from NIH-3T3, and PVA filature material has mechanical property and good cell compatibility.
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Objective To study the mechanism of protective effects of total rhizoma panacis japonica saponins(tRPJS)on the cerebral ischemia injury.Methods The middle cerebral artery occlusion model (MCAO)in rats and cerebral ischemia-reperfusion models in mice were used to investigate the influence of tRPJS on the nitric oxide synthase(NOS)and inducible nitric oxide synthase(iNOS)activity in hippocampus region.Results tRPJS significantly decreased the contents of NOS and iNOS in hippocampus region of MCAO rat and cerebral ischemia reinfusion mouse.Conclusion tRPJS has significantly protective effects by decreasing NOS and iNOS.
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BACKGROUND: Type Ⅱ collagen has been used as the carrier for chondrocyte transplantation in animal models, but whether type Ⅱ collagen may cause arthritis or mediate cytotoxicity remains unknown.OBJECTIVE: To detect the cellular immune functions of the New Zealand rabbits immunized by porcine type Ⅱ collagen.DESIGN: An exploratory comparative study based on the observations.SETTING: An institute of trauma surgery of a municipal hospital.MATERIALS: The study was conducted in the Institute of Trauma Surgery,Guangzhou Red Cross Hospital from August 1999 to February 2000. Six New Zealand rabbits, whose body mass ranged from 2.0 kg to 3.0 kg, were chosen of either gender.METHODS: The rabbits were immunized by porcine type Ⅱ collagen for 60days, during which the plasma was regularly taken for detection of type Ⅱ collagen antibody. On the 60th day, the peripheral blood as well as the spleens and lymph nodes were taken to separate the lymphocytes, which were subjected to secondary stimulation with type Ⅱ collagen in vitro to observe the reactive cell proliferation. The lymphocytes were randomly divided into two groups, and the first group was treated with phytohemagglutinin(PHA) of different concentrations to serve as the positive control, in which non-specific immunity was examined; The second group was treated with type Ⅱ collagen of different concentrations for examining specific immunity.peripheral blood lymphocytes of normal and immunized rabbits.RESULTS: On the 21st day, the titer of the antibody presented the first peak, and 40 days after the re-injection of the antigen the second peak appeared, which maintained for 20 days and then gradually descended. The lymphocytes of the normal rabbits proliferated in response to PHA stimulation but not to the first stimulation with the type Ⅱ collagen. The lymphocytes of the immunized rabbits exhibited significant proliferation upon stimulations with both PHA and type Ⅱ collagen. At the concentration of 25 mg/L, type Ⅱ collagen stimulation was sufficient to induce lymphocyte proliferation, the peak of which occurred when the collagen concentration reached 50 mg/L.CONCLUSION: Xenogenic type Ⅱ collagen at an adequate concentration may induce the increase of the type Ⅱ collagen antibody in immunized rabbits and proliferation of lymphocytes of the spleens and peripheral blood to cause cellular immune reaction and even immunological arthritis in relation to the transplantation.
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We have prepared a wound dressing made from chitosan and collagen. Its clinical curative effect was detected. Chitosan solution was put into purified collagen solution. Then, the solution became sponge by means of freeze drying, and it was subjected to a series of toxicology tests, including acute toxicity, stimulation test, allergic and hemolysis tests, as well as the clinical test of openning trauma in orthopedics. All of the results of toxicology tests were negative. The chitosan-collagen sponge could not only accelerate the speed of curing but also restrain the extravasate. Therefore, the chitosan-collagen sponge has good biocompatibility and clinical curative effect. It is a prospective security-biomaterial for medical use.