[Objectives]To establish a thin-layer chromatography(TLC)method for the determination of rubiadin-1-methyl ether in Yao Medicine Chuanlianzhu(Damnacanthus giganteus).[Methods]A silica gel G thin-layer plate was adopte...[Objectives]To establish a thin-layer chromatography(TLC)method for the determination of rubiadin-1-methyl ether in Yao Medicine Chuanlianzhu(Damnacanthus giganteus).[Methods]A silica gel G thin-layer plate was adopted for TLC.Petroleum ether(60-90℃)-chloroform-methanol-water(7:15:3:1)was used as the developing solvent and inspected under ultraviolet lamp(365 nm).The content was determined by Inertsil ODS-3 C 18 column(4.60 mm×250 mm,5μm),mobile phase:acetonitrile-0.2%phosphoric acid gradient elution,detection wavelength 277 nm,flow rate 1.0 mL/min,column temperature 30℃,injection volume 10μL.[Results]The spots of 10 Chuanlianzhu samples from different origins showed the same color at the same position as the control,and the spots were clear and specific.The injection volume of rubiadin-1-methyl ether showed a good linear relationship in the range of 2.90-145μg(R=0.9996).The average recovery rate of rubiadin-1-methyl ether in the low,medium and high dose groups of Yao Medicine Chuanlianzhu was 98.72%,and RSD=1.78%.[Conclusions]This method can effectively identify Yao Medicine Chuanlianzhu medicinal materials and accurately determine the content of rubiadin-1-methyl ether in the medicinal materials.It provides a scientific basis for the development and utilization of Yao Medicine Chuanlianzhu medicinal resources.展开更多
4,5-Dibromo-1-methyl-2-trichloroacetylpyrrole Ⅰ, prepared by the reaction of 1-methyl-2-trichloroacetylpyrrole with Br2 in 98.6 % yield, was condensed with methyl L-2-amino-3- phenylpropanoate to afford methyl 2-(4,...4,5-Dibromo-1-methyl-2-trichloroacetylpyrrole Ⅰ, prepared by the reaction of 1-methyl-2-trichloroacetylpyrrole with Br2 in 98.6 % yield, was condensed with methyl L-2-amino-3- phenylpropanoate to afford methyl 2-(4,5-dibromo-1-methylpyrrole-2-carboxmido)-3-phenylpropanoate Ⅱ in 90.8% yield. Crystal data for Ⅰ: monoclinic system, space group P21/c with a = 8.595(3), b = 10.900(4), c = 12.321(5) ,A°,β = 92.292(7)°, V = 1153.4(8)A°^3, Dc = 2.213 g/cm^3, F(000) = 728, CTH4Br2Cl3NO, Mr = 384.28, λ = 0.71073 A°, μ(MoKa) = 7.688 mm^-1, Z = 4, R = 0.0338 and wR2 = 0.0840 for 1963 observed reflections with I 〉 2a(I). Crystal data for Ⅱ: monoclinic system, space group P21 with a = 4.886(2), b = 15.921(8), c = 11.635(6) A°,β = 101.803(9)°, V = 885.9(7) A°^3, Dc = 1.665 g/cm^3, F(000) = 440, C16H16Br2N2O3, Mr = 444.13, λ = 0.71073 A°, μ(MoKa) = 4.590 mm^-1, Z = 2, R = 0.0335 and wR2 = 0.0837 for 3191 observed reflections with I 〉 2σ(I). The crystal structure reveals that compound Ⅱ forms the one-dimensional chain structure via the intermolecular hydrogen bonds of N(2)-H…O(1).展开更多
BACKGROUND: To date, a complete protein expression profile of the midbrain substantia nigra in a mouse model of chronic Parkinson's disease, induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP), does ...BACKGROUND: To date, a complete protein expression profile of the midbrain substantia nigra in a mouse model of chronic Parkinson's disease, induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP), does not exist. In addition, there are no reports of analysis of differential protein expression. OBJECTIVE: To separate and evaluate MPTP-induced differential protein expression through the use of proteomics in the substantia nigra of a mouse model of chronic Parkinson's disease. DESIGN: Randomized controlled animal study. SETTING: Department of Neurology, the First Affiliated Hospital, Chongqing Medical University. MATERIALS: Sixteen 8-10-week old, healthy, male, C57BL mice, weighing 20-25 g, and of clean grade, were provided by the Experimental Animal Center of Chongqing Medical University. The experimental animals were disposed according to ethical criteria. MPTP was provided by Sigma Company, USA; Pdquest 2D image analysis software and gelatum/irradiance image analysis system (ChemiDoc XRS) by Bio-Rad, USA; and Voyager DE-PROMALD1-TOF-MS mass spectroscopy analyzer by AB1 Company, USA. METHODS: This study was performed in Chongqing Neurological Laboratory between November 2006 and July 2007. Mice were randomly divided into model and control groups, with 8 mice in each group. Mice in the model group were received a subcutaneous injection of MPTP (25 mg&g), twice a week, for five successive weeks, to establish a chronic Parkinson's disease model. Mice in the control group received the same volume of a subcutaneous saline injection at the same time points. Mice were sacrificed by anesthesia to rapidly obtain the midbrain for protein separation of the substantia nigra. MAIN OUTCOME MEASURES: (1) 2-ED handbook (Bio-Rad Company) was referenced for two-dimensional electrophoresis, (2) PDQUEST8,0 analytical electrophoresis pattern was adopted to evaluate differential protein expression. (3) Peptide mass finger print map and data were retrieved on http://www.prospector.ucsf.edu to compare differential substantia nigral protein expression in the two groups. RESULTS: Two-dimensional gel electrophoresis of substantia nigra tissue indicated that there were 33 differential protein expressions between the two groups. Three new proteins were evaluated, including α -enolase, which exhibited regulated expression, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B. CONCLUSION: There are three proteins that exhibit differential expression in the substantia nigra- α -enolase, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B.展开更多
1-methyl-4-phenylpyridinium ion (MPP^+) induces endoplasmic reticulum stress and activates caspase-12 in PC12 cells, leading to neuronal apoptosis. However, the underlying molecular mechanism remains unknown. The p...1-methyl-4-phenylpyridinium ion (MPP^+) induces endoplasmic reticulum stress and activates caspase-12 in PC12 cells, leading to neuronal apoptosis. However, the underlying molecular mechanism remains unknown. The present study investigated the regulatory effects of nerve growth factor (Akt activator) and lithium chloride (glycogen synthase kinase-3β inhibitor) on the endoplasmic reticulum stress signaling pathway. The results revealed that MPP+ induced expression of Bip and C/EBP homologous protein. The upregulation of Bip and C/EBP homologous protein, as well as the decreased pro-caspase-12 level induced by MPP^+ were inhibited by pretreatment of the nerve growth factor or lithium chloride. These results suggest that the phosphatidylinositol 3 kinase-Aktglycogen synthase kinase-3β pathway is involved in MPP-induced endoplasmic reticulum stress.展开更多
Objective: To explore the protective effect of minocycline on the apoptosis of cellular parkinsonism models induced by MPP^+ . Methods: Using PC12 cells as the apoptotic model of dopaminergic neurons, MC and MPP^+...Objective: To explore the protective effect of minocycline on the apoptosis of cellular parkinsonism models induced by MPP^+ . Methods: Using PC12 cells as the apoptotic model of dopaminergic neurons, MC and MPP^+ were added into the culture medium of PC12 cells, and using MTr to assay the cell viability and metabolic state; The cells apoptosis was assayed by electrophoresis method and using flow cytometry FACS to assay the apoptosis ratio. Results: Added the MPP^+ to get the concentration of 10μmol/L, the cellular parkinsonism model of apoptosis had been prepared. The pre-treatment of MC ( 100/μmol/L) could significantly increase the PC12 cell viability. The apoptosis ratio of MC+MPP^+ group was significantly lower than that of MPP^+ group, but was still significantly higher than that of control group. Conclusion: MC may protect the cell apoptosis induced by MPP^+ to some extent.展开更多
Apoptosis and viability of PC12 cells following 1-methyl-4-phenylpyridinium ion (MPP+)-induced injury were monitored by flow cytometry, following Annexin V-propidium iodide double labeling, and 3-(4,5-Dimethylthia...Apoptosis and viability of PC12 cells following 1-methyl-4-phenylpyridinium ion (MPP+)-induced injury were monitored by flow cytometry, following Annexin V-propidium iodide double labeling, and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, respectively. The release of lactate dehydrogenase, superoxide dismutase activity and levels of malondialdehyde were determined by UV spectrophotometry. The changes in mitochondrial membrane potential and the intracellular concentration of calcium were determined by flow cytometry, and the activity of caspase-3 was monitored by western blot. According to cell viability and apoptosis studies, MPP+-induced apoptosis in PC12 cells was inhibited in the presence of 10 tJg/mL of Eleutheroside B Our results indicate that the neuroprotective effect of Eleutheroside B, following MPP+-induced apoptosis in PC12 cells, involves increasing the anti-oxidative stress capacity of cells, maintaining the high-energy state of mitochondrial membrane potential, reducing intracellular calcium concentration and inhibiting caspase-3 activity.展开更多
At present, pathogenesis and mechanism of Parkinson disease (PD) are still unclear. Animal models of PD are essential tools in studies on etiology and therapy and should mimic the chronic pathological process, histo...At present, pathogenesis and mechanism of Parkinson disease (PD) are still unclear. Animal models of PD are essential tools in studies on etiology and therapy and should mimic the chronic pathological process, histological characteristics and motor behavior dysfunction. In recent years, transgenic mice have been widely utilized to study the mechanism of PD, and it has become imperative that a PD mouse model of motor behavioral dysfunction be established. OBJECTIVE: To compare the behavioral and histochemical characters of two neurotoxic mice model induced with 6-hydroxydopamine (6-OHDA) or 1-methyl-4-phenyl-1, 2, 3, 6 -tetrahydropyridine (MPTP), and a better method to mimic Parkinson disease will be found out. DESIGN: Parallel experiment. SETTING: Laboratory of Molecular Genetics, Department of Medical Genetics, Shanghai Jiao Tohg University. MATERIALS: Sixty 129Sv/C57BL6J male wild mice, SPF grade, 8 - 12 weeks old, weighing 20 - 25 g, were provided by Experimental Animal Center, Shanghai Jiao Tong University. All the surgery operation was performed according to the rules of Shanghai Jiaotong University Animal Committee. METHODS: The experiment was carried out in the Laboratory of Molecular Genetics (National Key Laboratory), Department of Medical Genetics, Shanghai Jiao Ttong University from March to August 2006. ①Thirty-two male mice were randomly divided into control group and drug treatment group with 16 mice in each group. Surgery was carried out and 6-OHDA was administrated to substantia nigra pars compacta (SNpc) and nigra-striatum pathway according to the different parameters with intoxication apparatus. Saline was injected to the other 16 mice according to the same paradigm. 1 mg/kg apomorphine was injected intraperitoneally 2 weeks later after surgery to induce the imbalanced rotation behavior for 40 minutes. ②Twenty-eight mice were randomly divided into 4 groups with 7 in each group, including low-dose, moderate-dose, high-dose groups and negative control group. Then, mice in the drug treatment group were injected intraperitoneally with 5, 10 and 15 mg/kg MPTP for 9 successive days. In addition, mice in the control group were injected with the same volume of saline for 9 days. Pole test and stride length test were utilized to detect coordinative behavioral dysfunction. Mice were sacrificed 20 days after MPTP treatment, and histochemical staining of tyrosine hydroxynase (TH) was used to observe the loss of dopaminergic neuron in SNpc. MAIN OUTCOME MEASURES: ① Success ratio of each model establishment method; ② inducible asymmetric cycle behavior test 2 weeks after 6-OHDA injection; ③behavioral dysfunction in pole test and stride length, morphological changes in brain tissue. RESULTS: Totally sixty mice were used in this experiment and 3 mice were excluded because of the hypersensitivity or the clumsy reaction in motor behavioral detection before MPTP treatment, therefore, data was analyzed with the rest 57 mice. ① Lethal ratio: Three out of 16 mice died in striatum injection group and 5 out of 16 mice died in nigro-striatal pathway group. No mouse died in MPTP treatment groups. ② Locomotor behavior: No dysfunction of locomotor was found in 6-OHDA treatment groups. However, several motor behavioral dysfunction were start to present at the 4th day of MPTP injection. ③ Asymmetric cycle behavior: No asymmetric cycle was induced successfully two weeks after 6-OHDA surgery. Mice show hypersensitive behavior 10 minutes after apomorphine injection, which lasted for about 20 minutes. ④ Pole test: From the 4^th day of MPTP treatment, mice started to display coordinate dysfunction, such as climbing down along the pole in spiral, moving slowly with hesitation. Some mice could not grab the pole and slide down along the pole at 4th day post injection. Comparing with 0 dose control group, all the threedrug treatment groups show significant different dysfunction from the 4th day to the 20th day post injection (P 〈 0.01). ⑤ stride length test: Mice's stride length decreased, when treated with MPTP, and the mice in the high dose group displayed obviously. ⑥ Dopaminergic neuron stained with TH in nigra pars compacta: The results indicated that administrated MPTP (from low dose to high dose) by intraperitoneal cause chronic lesions on the dopaminergic neuron in the SNpc. CONCLUSION: PD mice models induced with 6-OHDA show high mortality ratio and no asymmetric cycle was found after apomorphine injection. However, injection of MPTP intraperitoneally can simulate the chronic pathway of PD, typical histological changes are found and stable motor behavioral dysfunctions are displayed.展开更多
Infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) into the right common carotid artery produced hemiparkinsonian syndrome on contralateral limbs in 5 rhesus monkeys. The hemiparkinsonian syndrome produce...Infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) into the right common carotid artery produced hemiparkinsonian syndrome on contralateral limbs in 5 rhesus monkeys. The hemiparkinsonian syndrome produced responded to madopar medication and the circling motion changed from toward the MPTP-treated side to away from the MPTP-treated side. Long term use of madopar developed a peak-dose dyskinesia of the face and limbs at the contralateral side. The toxic effect of MPTP was confirmed biochemically by reduction of nigrostriatal DA and histologically by degeneration of nigral neurons on the MPTP-treated side. It is concluded that this hemiparkinsonian monkey model will be of value in the elucidation of the neural mechanism underlying L-DOPA or DA agonists induced dyskinesia in Parkinson’s disease and in the search for newer methods of treatment which would produce less dyskinesia.展开更多
~3H-2-deoxyglucose (2-DG) autoradiographic technique was used to study the ef feets of a monoamine-oxidase-B (MAO-B) inhibitor deprenyl and the neurotoxin Ⅰ-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) on 2-D...~3H-2-deoxyglucose (2-DG) autoradiographic technique was used to study the ef feets of a monoamine-oxidase-B (MAO-B) inhibitor deprenyl and the neurotoxin Ⅰ-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) on 2-DG uptake in the mouse brain. Following MPTP intoxication, 2-DG uptake was increased in the substantia nigra and lo(?)us ceruleus. At the same time, obvious abnormal behavior of the animals was induced. In the mice pretreated with deprenyl, 2-DG uptake was similar to that of control animal. Ab normal behavior. though present, was significantly milder than in mice given MPTP alone. It is concluded that MPTP interferes with the glucose metabolism in the substantia nigra and locus ceruleus and induces remarkable abnormal behavioral syndrome of mice. These deleterious effects can be blocked by pretreatment with deprenyl.展开更多
By using 1-methyl-2-formyl-5 -Y-substituted pyrrole (4-nitrophenyl)hydrazones as a model for nitrogen-containing heterocyclic aromatic compounds, the emission wavelength [lambda(max(em))] values df their fluorescence ...By using 1-methyl-2-formyl-5 -Y-substituted pyrrole (4-nitrophenyl)hydrazones as a model for nitrogen-containing heterocyclic aromatic compounds, the emission wavelength [lambda(max(em))] values df their fluorescence spectra have been measured. Correlation results show that the Delta E-em values are mainly affected by polar effects, but spin-delocalizatin effects also exist.展开更多
2-Benzyl-7-butoxyl-9-isobutyl-1-methyl-β-carboline bromide(H-2-65) was synthesized by the reaction of Harmine with 1-iodobutane via N9-alkylation, demethyl and N2-quaternarization to obtain the new compound. The re...2-Benzyl-7-butoxyl-9-isobutyl-1-methyl-β-carboline bromide(H-2-65) was synthesized by the reaction of Harmine with 1-iodobutane via N9-alkylation, demethyl and N2-quaternarization to obtain the new compound. The results demonstrate that H-2-65 has more remarkable anticancer activities in vitro. The results of 1H NMR, 13 C NMR, DEPT, g COSY, g HSQC, g HMBC, MS, single-crystal X-ray diffraction and elemental analysis showed that the title compound crystallizes in the triclinic system, space group P1 with a = 9.545(5), b = 11.724(5), c = 11.839(6) , α = 77.530(6), β = 87.169(6), γ = 72.823(5)o, Z = 2, V = 1235.8(10)3, Dc = 1.294 g·cm-3, F(000) = 504, the final R = 0.0453, wR = 0.1262 and S = 1.044.展开更多
Dopamine cell bodies in the substantia nigra of the midbrain and with their terminals projecting to the neostriatum form the nigrostriatum and these dopamine neurons degenerate in Parkinson’s disease (PD). Based on m...Dopamine cell bodies in the substantia nigra of the midbrain and with their terminals projecting to the neostriatum form the nigrostriatum and these dopamine neurons degenerate in Parkinson’s disease (PD). Based on metabolic and func- tional specialization of the cell bodies versus the axon terminals, the level and disposition of dopamine, its metabolites and enzymes are different in both regions and are likely to be affected differently in PD. We examined changes in the midbrain dopamine system following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), to test the hypothesis that a predisposing/sensitization stage and a inducing/precipitating stage underlie PD. Pregnant mice were treated with a low dose of MPTP during gestation days 8 - 12 to model the predisposing/sensitization stage, by interrupting the fetal mid- brain dopamine system during its neurogenesis. For the inducing/precipitating stage, the 12-weeks offspring were ad- ministered MPTP. The prenatal-MPTP offspring appear normal, but midbrain dopamine, 3,4-di-hydroxy-phenyl-acetic- acid, 3-methoxytyramine, tyrosine-hydroxylase and L-aromatic-amino-acid-decarboxylase, were reduced by 49.6%, 48%, 54%, 20.9% and 25%. Postnatal-MPTP of 10, 20, 30 mg/kg administered to the prenatal-PBS vs prenatal-MPTP offspring reduced midbrain dopamine by 43.6%, 47.2%, 70.3% vs 85.4%, 89.1%, 95.2%;tyrosine-hydroxylase by 30%, 63%, 81% vs 30.7%, 70.4%, 91.4%;L-aromatic-amino-acid-decarboxylase by 0%, 2%, 40% vs 32%, 40%, 58%. The prenatal-MPTP may render the DA system sensitive by causing sub-threshold reduction of DA, its metabolites and en- zymes, enabling postnatal-MPTP to reduce dopamine above the 70% - 80% PD-inducing threshold. Thus, the study may produce a prenatal predisposing/sensitization and postnatal inducing/precipitation model of PD. It also indicates that some cases of PD may have a fetal basis, in which sub-threshold nigrostriatal impairments occur early in life and PD-symptoms are induced during aging by further insults to the dopaminergic system that would not cause PD symptoms in normal indi-viduals.展开更多
Sodium 3,5-bis(hydroxyimino)-1-methyl-2,4,6-trioxocyclohexanide C7H5N2NaO5 (I) has been isolated as the only product of the reaction of nitrosation of methylphloroglucinol. The structure of the titled compound has bee...Sodium 3,5-bis(hydroxyimino)-1-methyl-2,4,6-trioxocyclohexanide C7H5N2NaO5 (I) has been isolated as the only product of the reaction of nitrosation of methylphloroglucinol. The structure of the titled compound has been determined from single crystal X-ray diffraction data. The hydrated C7H5N2NaO52.5H2O crystallizes in the monoclinic space group C2/c, with a(?) 16.408(3);b(?) 12.446(3);c(?) 13.716(3);(o) 126.34(3). The planar organic anion exists in a triketo-dihydroxyimino form with the C–O and C–N distances from 1.220(2) to 1.271(2)?? and from 1.292(2) to 1.293?? respectively. In the IR spectrum of I, the sharp absorption band occurred at 1681 cm-1 due to C=O stretching indicating the strong H-interactions. The correlations of theoretical (DFT-B3LYP/aug-cc-pVDZ) and experimental UV-vis absorption spectra in neutral and alkaline ethanolic solutions showed the existence of hydroxyimino-nitroso tautomerism while ionization of I.展开更多
The transfer kinetics of phenol between aqueous phase and N,N di(methyl heptyl) acetaminde (N503) in kerosene has been studied using Lewis cell technique. The effects of the factors including the concentrations of p...The transfer kinetics of phenol between aqueous phase and N,N di(methyl heptyl) acetaminde (N503) in kerosene has been studied using Lewis cell technique. The effects of the factors including the concentrations of phenol in aqueous phase and organic phase, the concentration of N503 in organic phase, the acidity of aqueous phase, the stirring speed and the temperature on the rates of forward and backward extraction of phenol have been examined. The regularity of extraction rate has been obtained. According to experimental results, the rates of both forward and backward extraction of phenol might be controlled by diffusion process. The diffusion step of phenol from aqueous phase to interface for forward extraction and from interface to aqueous phase for backward extraction might be the rate controlling steps.展开更多
In the present work,the experimental and the theoretical spectroscopic properties of 1-Methyl-6-Nitro-1H-Benzimidazole were investigated.The FT-IR(400~4 000cm^(-1))and FT-Raman spectra(100~4 000cm^(-1))of 1-Methyl-6-N...In the present work,the experimental and the theoretical spectroscopic properties of 1-Methyl-6-Nitro-1H-Benzimidazole were investigated.The FT-IR(400~4 000cm^(-1))and FT-Raman spectra(100~4 000cm^(-1))of 1-Methyl-6-Nitro-1H-Benzimidazole in the solid phase were recorded.Also,experimental NMR and UV spectra of titled molecule were measured.To interpret the experimental data,geometric parameters,vibrational frequencies,NMR,UV spectra and NLO analysis of the optimized molecule were calculated using ab initio Hartree–Fock(HF)method and density functional theory(B3LYP)method with the 6-31++G(d,p)and 6-311++G(d,p)basis sets.Vibrational bands were assigned based on the potential energy distribution using the VEDA 4program.The theoretical results showed good agreement with the experimental values.展开更多
基金Supported by State Administration of Traditional Chinese Medicine High-level Key Discipline Construction Project of Traditional Chinese Medicine-Ethnic Minority Pharmacy (Zhuang Pharmacy) (zyyzdxk-2023165)General Scientific Research Program of Guangxi University of Chinese Medicine in 2020 (2020MS063)+4 种基金Key R&D Project of Guangxi Science and Technology Department (Guike AB21196057)Young Talent Cultivation Program of Guangxi International Zhuang Medicine Hospital (2022001)Funding Project of High-level Talent Cultivation and Innovation Team of Guangxi University of Chinese Medicine (2022A008)Guangxi Traditional Chinese Medicine Interdisciplinary Innovation Team Project (GZKJ2309)State Administration of Traditional Chinese Medicine"Twelfth Five-Year Plan"Key Discipline of Traditional Chinese Medicine (Ethnic Pharmacy)Zhuang Pharmacy.
文摘[Objectives]To establish a thin-layer chromatography(TLC)method for the determination of rubiadin-1-methyl ether in Yao Medicine Chuanlianzhu(Damnacanthus giganteus).[Methods]A silica gel G thin-layer plate was adopted for TLC.Petroleum ether(60-90℃)-chloroform-methanol-water(7:15:3:1)was used as the developing solvent and inspected under ultraviolet lamp(365 nm).The content was determined by Inertsil ODS-3 C 18 column(4.60 mm×250 mm,5μm),mobile phase:acetonitrile-0.2%phosphoric acid gradient elution,detection wavelength 277 nm,flow rate 1.0 mL/min,column temperature 30℃,injection volume 10μL.[Results]The spots of 10 Chuanlianzhu samples from different origins showed the same color at the same position as the control,and the spots were clear and specific.The injection volume of rubiadin-1-methyl ether showed a good linear relationship in the range of 2.90-145μg(R=0.9996).The average recovery rate of rubiadin-1-methyl ether in the low,medium and high dose groups of Yao Medicine Chuanlianzhu was 98.72%,and RSD=1.78%.[Conclusions]This method can effectively identify Yao Medicine Chuanlianzhu medicinal materials and accurately determine the content of rubiadin-1-methyl ether in the medicinal materials.It provides a scientific basis for the development and utilization of Yao Medicine Chuanlianzhu medicinal resources.
基金The project was supported by the National 863 Program of China (No. 2004AA628030), and Natural Science Foundation of Guangdong Province (No. 31920)
文摘4,5-Dibromo-1-methyl-2-trichloroacetylpyrrole Ⅰ, prepared by the reaction of 1-methyl-2-trichloroacetylpyrrole with Br2 in 98.6 % yield, was condensed with methyl L-2-amino-3- phenylpropanoate to afford methyl 2-(4,5-dibromo-1-methylpyrrole-2-carboxmido)-3-phenylpropanoate Ⅱ in 90.8% yield. Crystal data for Ⅰ: monoclinic system, space group P21/c with a = 8.595(3), b = 10.900(4), c = 12.321(5) ,A°,β = 92.292(7)°, V = 1153.4(8)A°^3, Dc = 2.213 g/cm^3, F(000) = 728, CTH4Br2Cl3NO, Mr = 384.28, λ = 0.71073 A°, μ(MoKa) = 7.688 mm^-1, Z = 4, R = 0.0338 and wR2 = 0.0840 for 1963 observed reflections with I 〉 2a(I). Crystal data for Ⅱ: monoclinic system, space group P21 with a = 4.886(2), b = 15.921(8), c = 11.635(6) A°,β = 101.803(9)°, V = 885.9(7) A°^3, Dc = 1.665 g/cm^3, F(000) = 440, C16H16Br2N2O3, Mr = 444.13, λ = 0.71073 A°, μ(MoKa) = 4.590 mm^-1, Z = 2, R = 0.0335 and wR2 = 0.0837 for 3191 observed reflections with I 〉 2σ(I). The crystal structure reveals that compound Ⅱ forms the one-dimensional chain structure via the intermolecular hydrogen bonds of N(2)-H…O(1).
基金the National Natural Science Foundation of China, No. 30370499
文摘BACKGROUND: To date, a complete protein expression profile of the midbrain substantia nigra in a mouse model of chronic Parkinson's disease, induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP), does not exist. In addition, there are no reports of analysis of differential protein expression. OBJECTIVE: To separate and evaluate MPTP-induced differential protein expression through the use of proteomics in the substantia nigra of a mouse model of chronic Parkinson's disease. DESIGN: Randomized controlled animal study. SETTING: Department of Neurology, the First Affiliated Hospital, Chongqing Medical University. MATERIALS: Sixteen 8-10-week old, healthy, male, C57BL mice, weighing 20-25 g, and of clean grade, were provided by the Experimental Animal Center of Chongqing Medical University. The experimental animals were disposed according to ethical criteria. MPTP was provided by Sigma Company, USA; Pdquest 2D image analysis software and gelatum/irradiance image analysis system (ChemiDoc XRS) by Bio-Rad, USA; and Voyager DE-PROMALD1-TOF-MS mass spectroscopy analyzer by AB1 Company, USA. METHODS: This study was performed in Chongqing Neurological Laboratory between November 2006 and July 2007. Mice were randomly divided into model and control groups, with 8 mice in each group. Mice in the model group were received a subcutaneous injection of MPTP (25 mg&g), twice a week, for five successive weeks, to establish a chronic Parkinson's disease model. Mice in the control group received the same volume of a subcutaneous saline injection at the same time points. Mice were sacrificed by anesthesia to rapidly obtain the midbrain for protein separation of the substantia nigra. MAIN OUTCOME MEASURES: (1) 2-ED handbook (Bio-Rad Company) was referenced for two-dimensional electrophoresis, (2) PDQUEST8,0 analytical electrophoresis pattern was adopted to evaluate differential protein expression. (3) Peptide mass finger print map and data were retrieved on http://www.prospector.ucsf.edu to compare differential substantia nigral protein expression in the two groups. RESULTS: Two-dimensional gel electrophoresis of substantia nigra tissue indicated that there were 33 differential protein expressions between the two groups. Three new proteins were evaluated, including α -enolase, which exhibited regulated expression, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B. CONCLUSION: There are three proteins that exhibit differential expression in the substantia nigra- α -enolase, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B.
基金the National Natural Science Foundation of China, No. 30860085a grant from the Candidates of Young and Middle-Aged Academic Leaders of Yunnan Province, No. 2006PY01-07the Natural Science Foundation of Yunnan Province, No. 2007C177M
文摘1-methyl-4-phenylpyridinium ion (MPP^+) induces endoplasmic reticulum stress and activates caspase-12 in PC12 cells, leading to neuronal apoptosis. However, the underlying molecular mechanism remains unknown. The present study investigated the regulatory effects of nerve growth factor (Akt activator) and lithium chloride (glycogen synthase kinase-3β inhibitor) on the endoplasmic reticulum stress signaling pathway. The results revealed that MPP+ induced expression of Bip and C/EBP homologous protein. The upregulation of Bip and C/EBP homologous protein, as well as the decreased pro-caspase-12 level induced by MPP^+ were inhibited by pretreatment of the nerve growth factor or lithium chloride. These results suggest that the phosphatidylinositol 3 kinase-Aktglycogen synthase kinase-3β pathway is involved in MPP-induced endoplasmic reticulum stress.
文摘Objective: To explore the protective effect of minocycline on the apoptosis of cellular parkinsonism models induced by MPP^+ . Methods: Using PC12 cells as the apoptotic model of dopaminergic neurons, MC and MPP^+ were added into the culture medium of PC12 cells, and using MTr to assay the cell viability and metabolic state; The cells apoptosis was assayed by electrophoresis method and using flow cytometry FACS to assay the apoptosis ratio. Results: Added the MPP^+ to get the concentration of 10μmol/L, the cellular parkinsonism model of apoptosis had been prepared. The pre-treatment of MC ( 100/μmol/L) could significantly increase the PC12 cell viability. The apoptosis ratio of MC+MPP^+ group was significantly lower than that of MPP^+ group, but was still significantly higher than that of control group. Conclusion: MC may protect the cell apoptosis induced by MPP^+ to some extent.
基金the Major Projects of National Science and Technology, No.2009ZX09103-329the National Natural Science Foundation for Distinguished Young Scholars of China, No.30901974+1 种基金Outstanding Youth Science Fund Program of Heilongjiang Province, No.JC200705"Spring Sunshine" Plan of Ministry of Education, No.2006
文摘Apoptosis and viability of PC12 cells following 1-methyl-4-phenylpyridinium ion (MPP+)-induced injury were monitored by flow cytometry, following Annexin V-propidium iodide double labeling, and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, respectively. The release of lactate dehydrogenase, superoxide dismutase activity and levels of malondialdehyde were determined by UV spectrophotometry. The changes in mitochondrial membrane potential and the intracellular concentration of calcium were determined by flow cytometry, and the activity of caspase-3 was monitored by western blot. According to cell viability and apoptosis studies, MPP+-induced apoptosis in PC12 cells was inhibited in the presence of 10 tJg/mL of Eleutheroside B Our results indicate that the neuroprotective effect of Eleutheroside B, following MPP+-induced apoptosis in PC12 cells, involves increasing the anti-oxidative stress capacity of cells, maintaining the high-energy state of mitochondrial membrane potential, reducing intracellular calcium concentration and inhibiting caspase-3 activity.
基金the grants from Shanghai Educational Committee, No. 03BZ03Department of Education for Doctor Foundation, No. 20040266014Shanghai Jiao Tong University Medical School for Doctor Foundation, No. BXJ0304
文摘At present, pathogenesis and mechanism of Parkinson disease (PD) are still unclear. Animal models of PD are essential tools in studies on etiology and therapy and should mimic the chronic pathological process, histological characteristics and motor behavior dysfunction. In recent years, transgenic mice have been widely utilized to study the mechanism of PD, and it has become imperative that a PD mouse model of motor behavioral dysfunction be established. OBJECTIVE: To compare the behavioral and histochemical characters of two neurotoxic mice model induced with 6-hydroxydopamine (6-OHDA) or 1-methyl-4-phenyl-1, 2, 3, 6 -tetrahydropyridine (MPTP), and a better method to mimic Parkinson disease will be found out. DESIGN: Parallel experiment. SETTING: Laboratory of Molecular Genetics, Department of Medical Genetics, Shanghai Jiao Tohg University. MATERIALS: Sixty 129Sv/C57BL6J male wild mice, SPF grade, 8 - 12 weeks old, weighing 20 - 25 g, were provided by Experimental Animal Center, Shanghai Jiao Tong University. All the surgery operation was performed according to the rules of Shanghai Jiaotong University Animal Committee. METHODS: The experiment was carried out in the Laboratory of Molecular Genetics (National Key Laboratory), Department of Medical Genetics, Shanghai Jiao Ttong University from March to August 2006. ①Thirty-two male mice were randomly divided into control group and drug treatment group with 16 mice in each group. Surgery was carried out and 6-OHDA was administrated to substantia nigra pars compacta (SNpc) and nigra-striatum pathway according to the different parameters with intoxication apparatus. Saline was injected to the other 16 mice according to the same paradigm. 1 mg/kg apomorphine was injected intraperitoneally 2 weeks later after surgery to induce the imbalanced rotation behavior for 40 minutes. ②Twenty-eight mice were randomly divided into 4 groups with 7 in each group, including low-dose, moderate-dose, high-dose groups and negative control group. Then, mice in the drug treatment group were injected intraperitoneally with 5, 10 and 15 mg/kg MPTP for 9 successive days. In addition, mice in the control group were injected with the same volume of saline for 9 days. Pole test and stride length test were utilized to detect coordinative behavioral dysfunction. Mice were sacrificed 20 days after MPTP treatment, and histochemical staining of tyrosine hydroxynase (TH) was used to observe the loss of dopaminergic neuron in SNpc. MAIN OUTCOME MEASURES: ① Success ratio of each model establishment method; ② inducible asymmetric cycle behavior test 2 weeks after 6-OHDA injection; ③behavioral dysfunction in pole test and stride length, morphological changes in brain tissue. RESULTS: Totally sixty mice were used in this experiment and 3 mice were excluded because of the hypersensitivity or the clumsy reaction in motor behavioral detection before MPTP treatment, therefore, data was analyzed with the rest 57 mice. ① Lethal ratio: Three out of 16 mice died in striatum injection group and 5 out of 16 mice died in nigro-striatal pathway group. No mouse died in MPTP treatment groups. ② Locomotor behavior: No dysfunction of locomotor was found in 6-OHDA treatment groups. However, several motor behavioral dysfunction were start to present at the 4th day of MPTP injection. ③ Asymmetric cycle behavior: No asymmetric cycle was induced successfully two weeks after 6-OHDA surgery. Mice show hypersensitive behavior 10 minutes after apomorphine injection, which lasted for about 20 minutes. ④ Pole test: From the 4^th day of MPTP treatment, mice started to display coordinate dysfunction, such as climbing down along the pole in spiral, moving slowly with hesitation. Some mice could not grab the pole and slide down along the pole at 4th day post injection. Comparing with 0 dose control group, all the threedrug treatment groups show significant different dysfunction from the 4th day to the 20th day post injection (P 〈 0.01). ⑤ stride length test: Mice's stride length decreased, when treated with MPTP, and the mice in the high dose group displayed obviously. ⑥ Dopaminergic neuron stained with TH in nigra pars compacta: The results indicated that administrated MPTP (from low dose to high dose) by intraperitoneal cause chronic lesions on the dopaminergic neuron in the SNpc. CONCLUSION: PD mice models induced with 6-OHDA show high mortality ratio and no asymmetric cycle was found after apomorphine injection. However, injection of MPTP intraperitoneally can simulate the chronic pathway of PD, typical histological changes are found and stable motor behavioral dysfunctions are displayed.
文摘Infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) into the right common carotid artery produced hemiparkinsonian syndrome on contralateral limbs in 5 rhesus monkeys. The hemiparkinsonian syndrome produced responded to madopar medication and the circling motion changed from toward the MPTP-treated side to away from the MPTP-treated side. Long term use of madopar developed a peak-dose dyskinesia of the face and limbs at the contralateral side. The toxic effect of MPTP was confirmed biochemically by reduction of nigrostriatal DA and histologically by degeneration of nigral neurons on the MPTP-treated side. It is concluded that this hemiparkinsonian monkey model will be of value in the elucidation of the neural mechanism underlying L-DOPA or DA agonists induced dyskinesia in Parkinson’s disease and in the search for newer methods of treatment which would produce less dyskinesia.
文摘~3H-2-deoxyglucose (2-DG) autoradiographic technique was used to study the ef feets of a monoamine-oxidase-B (MAO-B) inhibitor deprenyl and the neurotoxin Ⅰ-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) on 2-DG uptake in the mouse brain. Following MPTP intoxication, 2-DG uptake was increased in the substantia nigra and lo(?)us ceruleus. At the same time, obvious abnormal behavior of the animals was induced. In the mice pretreated with deprenyl, 2-DG uptake was similar to that of control animal. Ab normal behavior. though present, was significantly milder than in mice given MPTP alone. It is concluded that MPTP interferes with the glucose metabolism in the substantia nigra and locus ceruleus and induces remarkable abnormal behavioral syndrome of mice. These deleterious effects can be blocked by pretreatment with deprenyl.
文摘By using 1-methyl-2-formyl-5 -Y-substituted pyrrole (4-nitrophenyl)hydrazones as a model for nitrogen-containing heterocyclic aromatic compounds, the emission wavelength [lambda(max(em))] values df their fluorescence spectra have been measured. Correlation results show that the Delta E-em values are mainly affected by polar effects, but spin-delocalizatin effects also exist.
基金Supported by the National Key Technology Research and Development Program of the Ministry of Science and Technology of China(No.2012BAI30B00)
文摘2-Benzyl-7-butoxyl-9-isobutyl-1-methyl-β-carboline bromide(H-2-65) was synthesized by the reaction of Harmine with 1-iodobutane via N9-alkylation, demethyl and N2-quaternarization to obtain the new compound. The results demonstrate that H-2-65 has more remarkable anticancer activities in vitro. The results of 1H NMR, 13 C NMR, DEPT, g COSY, g HSQC, g HMBC, MS, single-crystal X-ray diffraction and elemental analysis showed that the title compound crystallizes in the triclinic system, space group P1 with a = 9.545(5), b = 11.724(5), c = 11.839(6) , α = 77.530(6), β = 87.169(6), γ = 72.823(5)o, Z = 2, V = 1235.8(10)3, Dc = 1.294 g·cm-3, F(000) = 504, the final R = 0.0453, wR = 0.1262 and S = 1.044.
文摘Dopamine cell bodies in the substantia nigra of the midbrain and with their terminals projecting to the neostriatum form the nigrostriatum and these dopamine neurons degenerate in Parkinson’s disease (PD). Based on metabolic and func- tional specialization of the cell bodies versus the axon terminals, the level and disposition of dopamine, its metabolites and enzymes are different in both regions and are likely to be affected differently in PD. We examined changes in the midbrain dopamine system following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), to test the hypothesis that a predisposing/sensitization stage and a inducing/precipitating stage underlie PD. Pregnant mice were treated with a low dose of MPTP during gestation days 8 - 12 to model the predisposing/sensitization stage, by interrupting the fetal mid- brain dopamine system during its neurogenesis. For the inducing/precipitating stage, the 12-weeks offspring were ad- ministered MPTP. The prenatal-MPTP offspring appear normal, but midbrain dopamine, 3,4-di-hydroxy-phenyl-acetic- acid, 3-methoxytyramine, tyrosine-hydroxylase and L-aromatic-amino-acid-decarboxylase, were reduced by 49.6%, 48%, 54%, 20.9% and 25%. Postnatal-MPTP of 10, 20, 30 mg/kg administered to the prenatal-PBS vs prenatal-MPTP offspring reduced midbrain dopamine by 43.6%, 47.2%, 70.3% vs 85.4%, 89.1%, 95.2%;tyrosine-hydroxylase by 30%, 63%, 81% vs 30.7%, 70.4%, 91.4%;L-aromatic-amino-acid-decarboxylase by 0%, 2%, 40% vs 32%, 40%, 58%. The prenatal-MPTP may render the DA system sensitive by causing sub-threshold reduction of DA, its metabolites and en- zymes, enabling postnatal-MPTP to reduce dopamine above the 70% - 80% PD-inducing threshold. Thus, the study may produce a prenatal predisposing/sensitization and postnatal inducing/precipitation model of PD. It also indicates that some cases of PD may have a fetal basis, in which sub-threshold nigrostriatal impairments occur early in life and PD-symptoms are induced during aging by further insults to the dopaminergic system that would not cause PD symptoms in normal indi-viduals.
文摘Sodium 3,5-bis(hydroxyimino)-1-methyl-2,4,6-trioxocyclohexanide C7H5N2NaO5 (I) has been isolated as the only product of the reaction of nitrosation of methylphloroglucinol. The structure of the titled compound has been determined from single crystal X-ray diffraction data. The hydrated C7H5N2NaO52.5H2O crystallizes in the monoclinic space group C2/c, with a(?) 16.408(3);b(?) 12.446(3);c(?) 13.716(3);(o) 126.34(3). The planar organic anion exists in a triketo-dihydroxyimino form with the C–O and C–N distances from 1.220(2) to 1.271(2)?? and from 1.292(2) to 1.293?? respectively. In the IR spectrum of I, the sharp absorption band occurred at 1681 cm-1 due to C=O stretching indicating the strong H-interactions. The correlations of theoretical (DFT-B3LYP/aug-cc-pVDZ) and experimental UV-vis absorption spectra in neutral and alkaline ethanolic solutions showed the existence of hydroxyimino-nitroso tautomerism while ionization of I.
文摘The transfer kinetics of phenol between aqueous phase and N,N di(methyl heptyl) acetaminde (N503) in kerosene has been studied using Lewis cell technique. The effects of the factors including the concentrations of phenol in aqueous phase and organic phase, the concentration of N503 in organic phase, the acidity of aqueous phase, the stirring speed and the temperature on the rates of forward and backward extraction of phenol have been examined. The regularity of extraction rate has been obtained. According to experimental results, the rates of both forward and backward extraction of phenol might be controlled by diffusion process. The diffusion step of phenol from aqueous phase to interface for forward extraction and from interface to aqueous phase for backward extraction might be the rate controlling steps.
文摘In the present work,the experimental and the theoretical spectroscopic properties of 1-Methyl-6-Nitro-1H-Benzimidazole were investigated.The FT-IR(400~4 000cm^(-1))and FT-Raman spectra(100~4 000cm^(-1))of 1-Methyl-6-Nitro-1H-Benzimidazole in the solid phase were recorded.Also,experimental NMR and UV spectra of titled molecule were measured.To interpret the experimental data,geometric parameters,vibrational frequencies,NMR,UV spectra and NLO analysis of the optimized molecule were calculated using ab initio Hartree–Fock(HF)method and density functional theory(B3LYP)method with the 6-31++G(d,p)and 6-311++G(d,p)basis sets.Vibrational bands were assigned based on the potential energy distribution using the VEDA 4program.The theoretical results showed good agreement with the experimental values.