Effect of Sinomenine on Expression of COX2, α7nAChR and A2A in A549 Cells / 广州中医药大学学报

Objective To observe the effect of sinomenine (SIN) on the expression of cyclooxygenase (COX2),alpha-7 nicotinic acetylcholine receptor(α7nAChR) and adenosine receptor(A2A) in A549 cells,and to explore the relative mechanism for cell proliferation.Methods The effect of SIN and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on the proliferation of A549 cells was determined by methyl thiazolyl tetrazolium (MTT) assay.The effect of SIN and NNK on the migration of A549 cells was detected by cell wound scratch assay.The effect of SIN and NNK on COX2 expression in A549 cells was determined by Western blotting method.The effect of SIN and NNK on the expression of α7nAChR and A2A mRNA and protein was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting method.Results NNK increased the proliferation and migration of A549 cells,while SIN inhibited the proliferation and migration of A549 cells.COX2 expression level was increased in NNK group but was decreased in SIN group.The expression levels of α7nAChR and A2A were up-regulated in NNK group but were down-regulated in SIN group.Conclusion SIN plays a role in inhibiting the proliferation and migration of A549 cells by suppressing COX2 expression.SIN has an inhibitory effect on the expression of α7nAChR and A2A.

Effect of Sinomenine on Expression of Purinergic Receptors A2A and P2X7 in Mouse Model and In-vitro Macrophages Stimulated by Lipopolysaccharide / 广州中医药大学学报

Objective To investigate the effect of sinomenine on the purinergic receptors A2A and P2X7 in endotoxemia mouse model and RAW264.7 macrophage model stimulated by lipopolysaccharide(LPS). Methods BALB/c mice were randomly divided into blank control group, model group and sinomenine group. Thirty minutes after the rats of sinomenine group were pretreated with intraperitoneal injection of sinomenine (40, 80, 160 mg/kg), the mice were given intraperitoneal injection of 15 mg/kg LPS to induce endotoxemia model. The serum levels of tumor necrosis factor-alpha(TNF-α) and interleukin-6(IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). The expression levels of purinergic receptor A2A and P2X7 in the liver and spleen were detected by reverse transcription-polymerase chain reaction(RT-PCR). RAW264.7 macrophages were divided into blank control group, LPS group and sinomenine group. Sinomenine group was firstly treated with sinomenine(300μmol/L) for 2 h, and then LPS group and sinomenine group were treated with LPS (100 ng/mL) for another 8 hours. TNF-α in the cell supernatant was measured by ELISA, and the expression levels of A2A and P2X7 in RAW264.7 cells were detected by RT-PCR. Results Stimulation with LPS could induce the increase of the mouse serum levels of TNF-α and IL-6 as well as the expression of A2A and P2X7 in mouse liver and spleen, and sinomenine had a counteraction on the above indexes(P<0.05) . In-vitro stimulation with LPS could induce the increase of the content of TNF-α and the expression of A2A and P2X7 in RAW264.7 cells , and sinomenine decreased TNF-α content and P2X7 expression (P<0.05) , but had an effect on enhancing A2A expression. Conclusion Sinomenine suppresses the expression of purinergic receptor P2X7 in mouse spleen and liver as well as in RAW264.7 macrophages, but its effect on the expression of A2A in various tissues and cells varies, whose related mechanism is needed further study.

Anti-inflammatory Effect of Three Kinds of Aconine from Radix Aconiti Lateralis Preparata on Macrophages / 广州中医药大学学报

Objective To observe the anti-inflammatory effects of three kinds of aconine from Radix Aconiti Lateralis Preparata ( Fuzi) on macrophages, as well as the effects of their combination with sinomenine. Methods The effect of three monoester alkaloids from Fuzi, benzoylaconine ( BAC) , benzoylmesaconine ( BMA) and benzoylhypaconine ( BHA) , on RAW264.7 cells proliferation was detected by cell counting kit 8 (CCK8) in vitro. RAW 264.7 macrophage cells were stimulated by lipopolysaccharides (LPS) and then treated with different concentrations of BAC, BMA, and BHA. Tumor necrosis factor alpha ( TNF-α) and interleukin 6 (IL-6) secreted by RAW264.7 cells were measured by enzyme-linked immunosorbent assay (ELISA) . The anti-inflammatory effects of three monoester alkaloids combined with sinomenine were evaluated by Zhengjun Jin Q method. Results The concentrations of TNF-α and IL-6 secreted by LPS-stimulated RAW264.7 cells were increased significantly ( P<0.01) , and then were inhibited by BAC ( 20, 30 μmol/L) , BMA ( 40, 80, 160μmol/L) and BHA ( 40, 80, 160 μmol/L) significantly ( P<0.01) . Antagonistic effects were present when 30 μmol/L of BAC or 160 μmol/L of BMA or 160 μmol/L of BHA was respectively used together with 100 or 300 μmol/L of sinomenine (Q<0.85). Conclusion The three kinds of monoester alkaloids from Fuzi exert anti-inflammatory effect on macrophages, and the effective dose of BAC is lower than that of BMA and BHA. The combination of BAC, BMA or BHA at the analyzed dosage with sinomenine has antagonistic effect.

Effect of Yirui Capsules on Serum Inflammatory Cytokines and Hepatic Alpha-7 Nicotinic Acetylcholine Receptor Expression in Hyperlipidemia Rats / 广州中医药大学学报

Objective To study the relationship between the inflammatory cytokine levels and the expression of hepatic alpha-7 nicotinic acetylcholine receptor (α7 nAChR) in hyperlipidemia rats and to investigate the intervention effect of Yirui Capsules ( mainly composed of Radix Salviae Miltiorrhizae, Fructus Crataegi, Rhizoma Alismatis, etc). Methods Fifty male SD rats were randomly divided into blank control group ( fed with conventional diet, N=10) and high-fat diet group (fed with high-fat diet, N=40). After feeding for 14 d, the forty rats in high-fat diet group were further randomly divided into model control group, and low-, middle-, and high-dose Yirui Capsules groups (140, 280, 560 mg/kg, respectively), 10 rats in each group and the treatment lasting 30 days. And then, the serum levels of lipids were detected, C-reactive protein (CRP) and tumor necrosis factor (TNF) -α in the serum were detected by enzyme-linked immunosorbent assay ( ELISA) , and the expression of hepatic α7 nAChR was analyzed by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR) . Results Yirui Capsules in different dosages could improve serum lipid levels in hyperlipidemia rats to various degrees. The contents of serum inflammatory cytokines CRP and TNF-α and the mRNA expression of hepatic α7nAChR were increased in the model control group as compared with those in the blank control group ( P<0.01). The contents of inflammatory cytokines CRP, TNF-αand the mRNA expression of hepatic α7nAChR were decreased in the three Yirui Capsules groups as compared with those in the model control group ( P<0.01). The results of immunohistochemical staining for α7nAChR expression were consistent with RT-PCR results. Conclusion The expression of hepatic α7nAChR is increased in hyperlipidemia rats. Yirui capsules are effective on decreasing inflammatory cytokine levels, improving lipid metabolism, and down-regulating α7nAChR expression.

Effect of sinomenine on mouse RAW264.7 macrophage cells line polarization induced by LPS or IL-4 / 中国免疫学杂志

Objective:To investigate sinomenine (Sinomenine,SIN) effect on RAW264.7 cells polarization to M1 or M2 phenotype induced by lipopolysaccharide (LPS) or interleukin-4 (IL-4) .Methods:RAW264.7 cells were induced to polarize to M1 by LPS ,and to M2 by IL-4.Sinomenine effects on LPS or IL-4 induced macrophages:TNF-αand IL-10 secretion induced by different condition were detected by Enzyme linked immunosorbent assay (ELISA);The expression level of mRNA of Arginase1(Arg-1),Nitric oxide synthase(iNOS),suppressor of cytokine signaling protein-2(SOCS2) and suppressor of cytokine signaling protein-3(SOCS3) of M1/M2 phenotypes were detected by real time PCR respectively.Results:Sinomenine inhibited the increase of TNF-αsecretion,iNOS and SOCS3 mRNA expression level induced by LPS.Sinomenine inhibited the increase of IL-10 secretion and Arg-1 mRNA expression level induced by IL-4,but SOCS2 mRNA expression level was not affected by Sinomenine.Conclusion: Sinomenine can inhibite the macrophage polarization to M1 and M2 induced by LPS and IL-4.Sinomenine plays a regulatory role on imbalance of M1/M2,and is conducive to maintain the dynamic balance.

Transformation of antimicrobial peptide fusion gene of cecropin B and rabbit NP-1 to Houttuynia cordata / 中国中药杂志

<p><b>OBJECTIVE</b>To transform the antimicrobial peptide fusion gene of cecropin B and rabbit NP-1(CN) into Houttuynia cordata to improve its antimicrobic capability.</p><p><b>METHOD</b>The fusion gene of CN designed and synthesized artificially was recombined with expression vector pBI121. The recombined vector was transformed to Agrobacterium tumefaciens LBA4404, by which CN gene was transformed to the explants of H. cordata. The transgenic regeneration plantlets were selected by kanamycin and rapid screening PCR. The transgenic plants were identified by PCR-Southern of genomic DNA and RT-PCR. The disease resistances were detected by antibacterial zone trail of leaf extracts to E. coli K12 and infection by Rhizoctonia solani.</p><p><b>RESULT</b>Gene of interesting CN was inserted into genomic DNA and expressed in transformed H, cordata, whose resistance to E. coli K12 and Rh. solani was stronger than that of the non-transformed control.</p><p><b>CONCLUSION</b>The fusion gene CN can improve antimicrobic capability of transformed H. cordata.</p>

Effects of Berberine Combined with Magnolol and Honokiol on the Formation of THP-1 Macrophage-derived Foam Cells / 中药新药与临床药理

Objective To study the effects and mechanism of berberine in combination with magnolol and honokiol on the formation of THP-1 macrophage-derived foam cells. Methods THP-1 macrophages were induced into foam cells by ox-LDL, and intracellular cholesterol in foam cells was determined by HPLC-MS; the concentration of tumor necrosis factor-α in the supernate of THP-1 macrophages was tested by ELISA and cell-surface CD36 level was assessed byflow cytometry. Results Combined treatment of berberine, magnolol and honokiol significantly reduced cholesterol con-tent in foam cells, also inhibited expression of tumor necrosis factor-α and CD36 in THP-1 macrophage. Conclusion Co-administration of berberine, magnolol and honokiol can inhibit the formation of foam cells probably through, down-regulating the level of tumor necrosis factor-α and CD36.

Effect of total polysaccharides of Sijunzi decoction on the proliferation of intestinal epithelial cell line IEC-6 of rats / 中国组织工程研究

BACKGROUND: In a series of recent studies it was demonstrated thatpolysaccharides play important roles in many physiologic and pathologicprocessions, such as infection, inflammation, inter-cell adherence and sig nal conduction, immune identification, cell proliferation and differentiation, as well as maintenance of cell structure and function. But the protectiveeffect of plant polysaccharides on gastrointestinal mucosa needs further re search. OBJECTIVE: To observe the effect of the total polysaccharides of SijunziDecoction (SJZD) (TPSJ) in different concentrations on the proliferation ofrat intestinal epithelial cell line IEC-6. DESIGN: Observational controlled trial. SETTING: Central Laboratory, Guangdong Hospital of Traditional ChineseMedicine. MATERIALS: ①Cell line: The IEC-6 of normal rats (Catalog No. RL 1592) was purchased from American Type Culture Collection (ATCC). IEC6 cells were originated mainly from intestinal crypt cells. ②Reagents anddrugs: DMEM medium, bovine insulin, gentamicin, fetal bovine serum (FBS) and DPBS were purchased from GIBCO Ltd. Cell proliferation kit(MTT) was purchased from Roche Ltd. Indomethacin was purchased fromSigma Company. SJZD was composed of Dangshen (Codonopsis pilosula),Baizhu (Atractylodes macrocephala), Fuling (Poria cocos) and Gancao (Glycyrrhizae uralensis), and these four drugs were in same ratio as Pharmacopoeia. The four herbs were boiled in water, extracted twice for 8 hours.Extract was combined, decompressed, concentrated, centrifugated with high speed to take out insoluble substance, put in glass paper to receive reverse lotic water dialysis for 2 hours. The final decoction was concentrated by heating followed by extraction with 80% ethanol. After overnight precipitation at room temperature and combination of sedimen, the total polysaccharide was obtained by deproteinating with the Sevag method.METHODS: ①The IEC-6 cell line was maintained in T-150 flasks with DMEM culture solution, and then put in CO2 incubator at 37 ℃, at saturated humidity, cultured at 0.05 volume fraction CO2, after being taken out from dry ice and defrosted rapidly in water-bath at 37 ℃. Flasks were incubated at 37 ℃ in 5% CO2· Stock cells were subcultured at a dilution of1:7 every 5-7 days and the medium was changed once every 2 days. The cells in passage 15-20 were used for testing. ②IEC-6 cells were inoculated at a density of 1×l04 cells/well in 96-well plates. Cultured were supplemented with TPSJ in a final concentrations ranging from 50, 100 and 200 mg/L after 6 hours, which was 3 TPSJ groups. One plate would be taken out for the examination of cell proliferation using MTT assay everyday. The cells that not administrated by any intervention were used as normal control group and cell proliferation was assayed using MTF at corresponding time points. ③IEC-6 cells were inoculated at a density of1×104 cells/well in 96-well plates, and then cultured in the DMEM supplemented with no serum from the following day for 24 hours. For the examiation of mucosal restitution, indomethacin at concentration of 40 mmol/L was employed to induce IEC-6 cells injured, which was indomethacin group. The three concentration of TPSJ was 50, 100 and 200 mg/L, respectively, which was 50,100,200 mg/L TPSJ groups. After drug action for 20 hours, the proliferation of cells was measured using MTT according to the manufacturer's instructions. IEC-cells without any intervention were used in the normal control group. Cell proliferation was determined with TT method at corresponding time points.MAIN OUTCOME MEASURES: MTT assay was used to examine the effects of TPSJ on IEC-6 cell proliferation in different times. MTT assay was used to detect the effect of TPSJ on IEC-6 cell proliferation inhibited by indomethacin.RESULTS: TPSJ could accelerate IEC-6 cells growth at different doses and in different time. After the cells were treated by 40 mmol/L indomethacin for 24 hours, the absorbance (A) of IEC-6 cells apparently declined compared with that in the normal control group (0.17±0.02,0.31±0.03; P ＜ 0.01). The A of IEC-6 cells treated by TPSJ in 100 mg/L group was apparently higher compared with indomechacin group (0.25±0.04, 0.17±0.02; P ＜ 0.01). The A of IEC-6 cells treated by TPSJ did not restored to the normal level, but there was no insignificant difference compared with normal group (P ＞ 0.05).CONCLUSION: TPSJ can accelerate the proliferation of IEC-6 cells. TPSJ can exert regulatory function both in intestinal mucosa absorption and immunity by affecting intestinal epithelial cells.

Effect of angong niuhuang wan on serum lactate dehydrogenase isoenzymes in rats with cerebral infarction / 中国组织工程研究

BACKGROUND: Angong niuhuang wan consists of mercury and arsenic, which has drawn the attention on its safety. It is necessary to carry on the research on evaluation in its availability and safety.OBJECTIVE: To study the mechanic differences of angong niuhuang wan in the organic body physiologically and pathologically.DESIGN: Randomized controlled experiment based on the experimental animals.SETTING: Institute of clinical pharmacology affiliated to a university.MATERIALS: The experiment was performed in Institute of Clinical Phar macology affiliated to Guangzhou University of Traditional Chinese Medicine from March to April in 2001. Guangdong Medical Experimental Animal Center provided 24 SD male rats, weighted varied from 250 g to 300 g.METHODS: SD rats were randomized into 4 groups, named as normal group, normal & angong niuhuang wan group(normal & wan group), model of cerebral infarction group(model group) (The middle cerebral artery embolism was induced in rats by photochemistry. ) and model of cerebral in farction group & angong niuhuang wan group (model & wan group), 6 rats in each group. Medication instruction: gastric feeding was applied once daily, 0. 13 g/kg, totally for 7 days. MAIN OUTCOME MEASURES: contents of lactate dehydrogenase (LDH) isoenzymes, LDH1-5.RESULTS: Contents of serum LDH1-3 in normal & wan group were significantly increased than those in normal group( P ＜ 0.01 ), of which, the values of LDH1, LDH2, LDH3 were(17.02 ±0.46)%, (14.70 ±0. 18)%,(15.47±0.13)% successively in normal & wan group, were(11.25 ± 0. 70) %, (8.26 ± 0. 90) %, ( 12. 86 ± 0.90) % successively in normal group. Content of serum LDH3[(15.51 ±2.60)% [ in model & wan group was significantly increased compared with that in model group[(10.93±2.10)% ] (P ＜ 0.01) . The contents of LDH4 were[(22.62 ±3.00)% ] and(28. 18 ±0. 80)% respectively in two groups, indicating significantly reducing( P ＜ 0.01 ).CONCLUSION: The organic injury of angong niuhuang wan is less in pathological state of focal cerebral infarction than that in physiological state,suggesting that the mechanic differences of angong niuhuang wan in organic body present in physiological and pathological states.

Overall alkali in tongbiling for proliferation of lymphocytes and transferrin receptor of T lymphocytes / 中国组织工程研究

BACKGROUND: Researches on the pathogenesis and pathological changes of rheumatoid arthritis(RA) have achieved significant progress in recent years. But traditional Chinese medicine(TCM) has unique advantage in RA therapy.OBJECTIVE: To study effects of overall alkali in tongbiling(TBL) on the proliferation of lymphocytes and the transferrin receptor of T lymphocytes (CD71) to explore the mechanism of TBL on the modulation of cell immunity.DESIGN: A completely randomized grouping design and an explorative study by employing cells as subjects.SETTING: Sixth internal medicine department of a TCM university,center of tissue transplantation and immunology college of life science of a university PARTICIPANTS: The study was conducted in the central laboratory (tertiary laboratory of National TCM Administrator) of the first affiliated hospital of Guangzhou TCM medical university between July 2002 and August 2003. Ten clean male SD rats were selected.METHODS: Lymphocyte was separated from rat inguinal lymph node for culture. Concanavalin(ConA) was used for 72-hour stimulation. The impacts of overall alkali TBL on lymphocyte proliferation were tested by MTT.The expression of T lymphocyte CD71 was tested by flow cytometer after 48-hour stimulation of phorbol 12,13 -dibutyrate(PDB) or ConA.MAIN OUTCOME MEASURES: The impacts of overall alkali TBL on lymphocyte proliferation and T cell activation.RESULTS: Different concentration of overall alkali TBL could significantly inhibit the proliferation of lymphocytes under ConA stimulation. PDB and ConA-activated T lymphocyte CD71 + expressions were significantly higher than that of blank control group(P<0.01) . CD3+ CD71 + expressions [(62.03±1.51) %,(25.28±1.57) %,(20. 29±1.72)%] activated by ConA under different concentration of overall alkali TBL(50,100,200 mg/L)were significantly lower than(72.03±1.28)% of BPS-positive control group (P<0. 05). CD3 + CD71 + expressions activated by PDB under 100 mg/Land 200 mg/L of overall alkali TBL were significantly lower than that of phosphate buffer solution (PBS-)positive control group(P<0.05). Different concentration of overall alkali TBL had significant down-regulated effects on CD71 expression in T lymphocyte activated by PDB or ConA and there was also a significant dose-effect relationship(P<0. 05). The inhibition on ConA-activated CD71 expression was stronger than that of PDB.CONCLUSION: Overall alkali TBL can inhibit the abnormal proliferation of T lymphocyte and its mechanism might be realized through its inhibition on transferrin receptor.

Effects of emodin on IL-8 secretion and NF-?B activation of HT-29 cells / 中国药理学通报

Aim To study the effect of emodin on IL-8 secretion and NF-?B activation of HT-29 cells,and explore the molecular mechanism of emodin.Methods The cytotoxicity of emodin was assessed by WST;NF-?B activation was detected with co-focal microscopy by immunofluorescence;the production of IL-8 was investigated by ELISA.Results Emodin with the concentration of 10~80 ?mol?L-1 could decrease the mass production of IL-8 Secretion of HT-29 cells stimulated by IFN-?+LPS in a dose-dependent manner.Emodin with various concentrations could inhibit NF-?B activation dose-dependently.Conclusions Emodin inhibited IL-8 secretion and NF-?B activation of HT-29 cells stimulated by IFN-?+LPS.

Inhibition of clotrimazole on Paneth cell defensins / 中国药理学通报

Aim To investigate the Clotrimazoles (CLT) inhibitive effect on the secretion of ?-defensins of the Paneth cells.Methods The ?-defensin mRNA expression is determined by semi-quantitative RT-PCR. The secretion of defensins is determined by dot-blot and ELISA. Results CLT has no influence on the defensin mRNA expression. All of 125 nmol?L -1, 250 nmol?L -1 and 500 nmol?L -1 CLT can inhibit the secretion of ?-defensins. Conclusion CLT could inhibit the secretion of defensins, which suggests that CLT maybe influence the function of small intestine

Studies on In-Vitro Biological Activity of Albiflorin R1 in Counteracting Cell Proliferation / 广州中医药大学学报

[Objective] To study the in-vitro biological activity of albiflorin R1 (AR1), a new monoterpene glycoside from Paeonia lactiflora,in counteracting cell proliferation. [Methods] Two kinds of human cancer cell strains (HeLa cells and K562 cells) and J774 macrophage cell strain were cultured in vitro to observe the effects of albiflorin Rl on cell proliferation. [Results] Albiflorin Rl could markedly inhibit the proliferation of J774 macrophages and also inhibit the proliferation of HeLa and K562 cell stains in vitro. [Conclusion] Albiflorin Rl may have an anti-neoplastic, anti-inflammatory and immunosuppressive activity.

Regulatory Effects of Total Alkaloids of Tongbiling Prescription on Cellular Immunity in Rats / 广州中医药大学学报

[Objective]To observe the effects of total alkaloids (TA) of Tongbiling (TBL) prescription on proliferation of T lymphocytes and transferring receptor of T lymphocytes (CD71) in rats and to explore the mechanism of TBL in regulating cellular immunity. [Methods]Isolated lymphocytes from mesenteric lymph node were cultured with various concentrations of TBL TA and canavaline A (ConA) for 72 hours; the proliferation of T lymphocytes was observed by MTT method and CD71 expression rate of T lymphocyte activated by phorbol 12,13-dibutyrate (PDB) or ConA for 48 hours was detected by flow cytometry.[Results]Various concentrations of TBL TA inhibited the proliferation of T lymphocytes activated by ConA and decreased CD71 expression rate of T lymphocyte activated by ConA or PDB in a dose-effect manner. Inhibitory effect of TBL TA on CD71 expression rate of T lymphocyte activated by ConA was superior to that activated by PDB. [Conclusion]TBL TA can inhibit the abnormal proliferation of T lymphocytes and its mechanism may be related to the expression of CD71.

Selective Inhibitory Effect of Sinomenine on Activity of Cyclooxygenase 2 / 广州中医药大学学报

[Objective] To explore the anti - inflammatory mechanism of sinomenine, a monomer alkaloid from Caulis Sinomenii. [ Methods] In - vitro enzyme was applied to observe the effect of sinomenine on cyclooxygenase (COX) 1 and COX 2. [Results] Sinomenine at the concentration of 1 - 250 ?mol/L could inhibit the formation of PGE2 induced by COX 2 in a good dose - effect manner. [Conclusion] Sinomenine selectively inhibiting the activity of COX 2 may be the anti - inflammatory and anagesic mechanism of sinomenine with less gastrointestinal adverse effect.

Effect of removing stasis medicinal herbs on proliferation and proteomics of THP-1 cell line induced by endotoxin / 中国免疫学杂志

Objective:To observe the proteomics of blood-activating and stasis-removing(BASR) herbs in cellular model of endotoxin infection in vitro.Methods:Cellular model of endotoxin was induced by LPS,and a technical system by two-dimensional electrophoresis was widely used,proteins in different expression were analyzed by PDQust 7.1.1.Results:There were 176 protein spots in model group,in which 26 protein spots with different expression were sieved.Compared the protein profiles of TanshinoneⅡA group in model group,proteins were found as two-way-regulated.Among them,50% proteins were observed down-regulated and 38.5% proteins were seen up-regulated in TanshinoneⅡA group.11.5% more up-regulated.Conclusion:Cellular model of endotoxin induce was applied to sieving traditional Chinese medicines in laboratory;The Tanshinone ⅡA can inhibit the proliferation of THP-1 cell line in infection model,and can modify expression of some proteins.

Effects of Angong Niuhuang Wan on Amounts of Isoenzymes of Serum Lactate Dehydrogenase in Rats with Cerebral Infarct / 中药新药与临床药理

Objective:To investigate the different effects of Angong Niuhuang Wan on normal and pathological body. Methods:Cerebral infarct rat models were established by photochemically initiated thrombosis. The rats were randomly allocated to four groups. The control group and the model group were treated with 0.9%NaCl for seven days, and the control+drug group and the model+drug group with Angong Niuhuang Wan 0.13g/kg for seven days. Then amounts of idoenzymes of serum lactate dehydrogenase(LDH1～5) were determinated by the method of gel electrophoresis. Results:The amounts of LDH1 and LDH2 in the model group increase obviously as compared with the control group(P

Identification of the Fruit of Terminalia chebula from Its Comfusable Varieties by RAPD Analysis / 中草药

Different varieties of Terminalia chebula Retz. were identified by DNA finger prints, andat the same time, some problems encountered during the application of RAPD for the identification ofTCM were studied. The polymorphic features of DNA finger prints of different species were compared.Results of the study showed that they were differently related in heredity which can provide a basis for thedifferentiation of one species from the other. Means for the solution of problems arised during RAPD ap-plication were suggested. It was concluded that RAPD is an effective method to identify T. chebula from itsconfusable varieties.

Study on Quality Standard of Jinyou Tankeqing Tablets / 中药新药与临床药理

Objective To study the quality standard of Jinyou Tankeqing Tablets (JTT). Methods Total flavonoids in JTT were identified by TLC. Naringin content in JTT was determined by HPLC. Results The relevant spots of total flavonoids in JTT can be identified by TLC. The content of naringin in JTT can be determined by HPLC. The linearity of naringin was good in the range of 2～20 ?g (r= 0.999 9). The average recovery of naringin was 101.29 %with RSD = 2.58 %. Conclusion The established quality standard is simple,feasible and repeatable,and can be used for the quality supervisory of JTT.

Isolation of Rat Marrow Mesenchymal Stem Cells and Effect of Conditioned Medium on Its Proliferation / 广州中医药大学学报

【Objective】To establish a simple,rapid and practical method for the in-vitro isolation,purification and proliferation of rat marrow mesenchymal stem cells(MSCs).【Methods】Rat MSCs were isolated and purified by differential adhesion method.The effects of conditioned medium on the morphologic characteristics and growth of rat MSCs were observed.【Results】Rat MSCs were effectively purified by differential adhesion method,and grew well with good proliferation and normal morphology in conditioned medium.【Conclusion】Differential adhesion method and conditioned medium are effective for the purification and culture of rat MSCs.