Objective To investigate the extraction method of total flavonoids from the leaves of ficus lacor and the protec tive effects of extraction on the cellular damage to provide a basis for the research on the phamaceutical value of ficus lacor leaves.Methods The ethanol extraction method was adopted to extract the total flavonoids in the leaves of ficus lacor and the extraction efficiency was calculated with rutin as the standard.The rotenone induced human lung adenocarcinoma cellular damage served as the model,then the influencesof the extraction on the cellular viability,cellular morphology,production of reactive oxygen species (ROS) and apoptosis were researched.Results The extraction efficiency of total flavonoids in the leaves of ficus lacor by 60％ ethanol was 5.02％;the extraction at the concentration of 32 mg/L could significantly inhibit the decrease of cell viability,cellular shape change,ROS production and apoptosis of A549 cells induced by 100μg/L rotenone.Conclusion The ethanol extraction method can be used to extract the total flavonoids in the leaves of ficus lacor and the extraction has the protective effects on the A549 cellular dam age induced by rotenone,the leaves of ficus lacor have the potential for further researching its pharmaceutical value.

The dissolution of substrate is the key step in steroid microbial conversion.The formation of surfactant micelle with a hydrophobic center directly results in increased pseudo-water-solubilities of hydrophobic compounds such as steroids.Results showed increased water solubility of steroid with the increased surfactant concentrations.Compared to mono-surfactant PSE,less quantity of multiple surfactants PSE-MGE was required to achieve the enhancement of steroid solubility at the same level,thus the negative effects of surfactants on mycelial growth were reduced and steroid conversion was therefore improved more.It was also shown that the surfactants and ultrasonic irradiation used jointly could result in better substrate dispersion and dissolution than surfactants used alone,and the best result was obtained when the comparatively high intensity (12.4 W/cm 2) and short time (5min) of ultrasonic irradiation were performed.

Objective To study the relationship between the MRI features and pathological types of intracranial meningiomas. Methods MRI findings of intracranial meningiomas in 63 cases proved by operation and pathology were analyzed retrospectively.Results Of 63 cases,62 cases were singular and one case was multiple lesions. The lesions were located in frontal,parietal and occipital regions nearby the convexity of the brain in 30 cases,in sellar region in 10,at sphenoidal crest in 8 , at olfactory sulcus in 5 and at other regions in 10. Isointense or slightly hypointense signal on T 1WI was found in 87.27% cases of meningiomas . On T 2WI, tumors had isointense or slightly hyperintense signal in 69.84%, obviously hyperintense signal in 19.05%,heterogenous texture signal in 7.94% and slightly low intense signal in 3.17%. Pathological type was included meningiomas meningothelial(n=29), meningiomas fibrous(n=11) , meningiomas psammomatous(n=8),angioblastic meningiomas (n=5) and meningiomas angiomatous(n=10). Conclusion It is great value in diagnosis of meningiomas with MRI. Meningiomas angiomatous show effecting of blood-vessel flowed empty with MR imaging.

Objective:To study the expression of osteoprotegerin(OPG)and receptor activator nuclearfactor kappa B ligand(RANKL)at protein level in human periodontal ligament cells(HPDLCs),and theeffect of 1?,25(OH)_2 vitamin D_3[1,25(OH)_2 vitD_3] on the secretion of OPG protein in vitro.Meth-ods:HPDLCs were harvested in vitro by sequential digestion with trypsin and collagenase.The expressionof RANKL in HPDLCs at protein level was tested by immunocyto-chemistry.Enzyme-linked immuno-adsordent assay(ELISA)was used to detect the OPG protein which was secreted into the culture mediumby HPDLCs cultured with and without 10~(-8) mol/L 1?,25(OH)_2 vitD_3 on the 0,2nd,4th,and 6th days,respectively.Results:RANKL protein was detected on the membrane and plasma of HPDLCs,and OPGprotein was secreted in the culture medium.The secretion of OPG protein was down-regulated by 10~(-8)mol/L 1?,25(OH)_2 vitD_3.Conclusion:HPDLCs have the bone metabolism system of OPG/RANKL,which works during the process of 1?,25(OH)_2 vitD_3 inducing HPDLCs.The conclusion has laid thegroundwork for the study on bone remodelling mechanisms of HPDLCs.

Objective To explore the value of monitoring in vertebral and spinal cord operation by the real-time monitoring of the spinal cord in the process of operation.Methods Nineteen cases were included by the real-time monitoring of spinal cord in the process of operation.Japanese Orthopaedic Association(JOA) score was used to evaluate the clinical efficacy during following-up before and discharge after operation,6 months and 12 months after operation.Results Intraoperative ultrasound monitoring in the 19 patients had no symptom of spinal cord injury and other complications within the plant without loosening fracture occurred were bony fusion; the average JOA scores in the patients before operation,discharge after operation,6 months and 12 months after operation were 8.80±1.60,14.00±1.57,14.60±1.61 and 14.80±1.58,respectively.The improvement rate of JOA score for discharge after operation,6 months and 12 months after operation were individually 51.6%,61.3%,and 64.5%.The JOA score after operation was obviously higher than that before operation.The significant difference between the two groups was observed in the experiment (P<0.05).The cervical spinal canal sagittal diameter significantly increased after operation (mean sagittal diameter of the expansion of 6.6 mm),compared with that before the trial (P<0.05).Conclusion The real-time monitoring with ultrasonography during operation can identify the change of spinal cord and provide the image evidence of operative efficiency to avoid the injury in vertebral and spinal cord operation.

OBJECTIVETo classify NZB/W F1 lupus mice into three different constitutions, i. e. the cold, normal, and hot constitutions, and to prove the objective existence of the difference among them.

METHODSUsing the Four Diagnosis Work Station for Mice (founded by Prof. FANG Zhao-qin, the Research Faculty of Experimental Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine), the body weight, the armpit temperature, the heart rate, the 35-s activities, and the color of their tails and claws (r value) were detected. The total weight was calculated according to the formula: The total weight sum = the correction value of claws x 1 + the correction value of tail x 0.2 + the correction value of the armpit temperature x 0.7 + the correction value of heart rate x0. 05 + the number of the quadrant crossing x 0.025. The NZB/W F1 lupus mice were classified into the three different constitutions according to the higher total weight sum, the hotter the constitution, the lower total weight sum, the colder the constitution. The hydroxyproline content, connective tissue growth factor (CTGF), and transforming growth factor-beta1, (TGF-beta1) gene expression difference in the renal tissue were detected and the immunofluorescence staining observed.

RESULTSAmong the 158 NZB/W F1 lupus mice, 33 mice were classified into the hot constitution, 34 into the cold constitution, and 91 into the normal constitution. The renal hydroxyproline content in mice of normal and cold constitutions were higher than that of mice of the hot constitution (P<0.01). No statistical difference was shown between mice of the normal constitution and mice of the cold constitution. The CTGF gene expression level was significantly higher in mice of the cold constitution and mice of the hot constitution than in mice of the normal constitution (P<0.05). No significant difference was found between mice of the cold constitution and mice of the hot constitution. Lower level of TGF-beta1, expression existed in mice of the cold constitution than in mice of the normal constitution or mice of the hot constitution, showing insignificant difference. The immunofluorescence stain of the renal tissue among the three constitutions also showed some difference.

CONCLUSIONConstitution difference did exist among NZB/W F1 lupus mice.

Animals ; Body Constitution ; Disease Models, Animal ; Female ; Kidney ; metabolism ; pathology ; Lupus Erythematosus, Systemic ; metabolism ; pathology ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred NZB ; classification

OBJECTIVETo explore Akt-regulated direct p53 mitochondrial translocation in cisplatin-induced apoptosis in ovarian cancer cells and the relationship between this and chemoresistance in ovarian cancer.

METHODSChemosensitive ovarian cancer cell lines (OV2008 and A2780s) and chemoresistant cells (C13(*) and A2780cp) were treated with cisplatin and whole cell and mitochondrial p53 contents were determined by Western blot. The p53 accumulation in mitochondria was determined in purified mitochondrial fractions in cisplatin-sensitive and -resistant ovarian cancer cells. Akt1/2 siRNA were transfected into C13(*) cells. Cisplatin-induced apoptosis was measured by Hoechst staining and p53 translocation was determined by Western blot.

RESULTSCisplatin induced mitochondrial p53 accumulation and apoptosis in chemosensitive cells (P < 0.05), but not in resistant cells (P > 0.05). Over-expression of active Akt2 inhibited p53 directly translocate to mitochondria, and downregulation of Akt by Akt1/2 siRNA increased p53 mitochondrial accumulation and sensitize C13(*) cells to cisplatin treatment.

CONCLUSIONSCisplatin induces direct p53 mitochondrial accumulation in chemosensitive cells, and Akt confers resistance in ovarian cancer cells, in part, by regulating the direct action of p53 in mitochondrial death pathway.

Antineoplastic Agents ; metabolism ; Apoptosis ; drug effects ; Blotting, Western ; Cell Line, Tumor ; Cisplatin ; metabolism ; pharmacology ; Drug Resistance, Neoplasm ; drug effects ; Female ; Humans ; Mitochondria ; genetics ; Ovarian Neoplasms ; drug therapy ; Proto-Oncogene Proteins c-akt ; genetics ; physiology ; RNA, Small Interfering ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Objective To evaluate the value of serum creatinine(Scr),creatinine clearance (Ccr),MDRD equation 7(MDRD 7),~(99)Tc~m-DTPA renal dynamic imaging(gGFR)and cystatin C in screening changed glomerular filtration function in type 2 diabetic patients.Methods The ~(99)Tc~m-Diethylene Triamine Pentaacetic Acid(~(99)Tc~m-DTPA)plasma clearance(rGFR)obtained with the dual plasma sampling method was used as a reference with which Scr,Ccr,MDRD 7,gGFR and Cystatin C were compared.Results Sixty of type 2 diabetic patients were selected,including 35 male and 25 female.The average age was(62.4?11.7)years and the average diabetic history was(10.66?9.35)years.Scr,Ccr, MDRD 7,gGFR were all correlated significantly with rGFR.Correlation coefficients were 0.675 for Ccr, -0.588 for Cystatin C,-0.500 for Scr,O.428 for MDRD 7,0.367 for gGFR(P values all

Toxicity of different processed was evaluated Polygoni Multiflori Radix by determining the hepatotoxic potency for selecting processing technology. Process Polygoni Multiflori Radix using high pressure steamed, Black Bean high pressure steamed, atmospheric steamed for different time. Using normal human hepatocytes (L02) as evaluation model, hepatotoxic potency as index to evaluate hepatotoxic potency of different processed Polygoni Multiflori Radix. Analysis chemical composition of some processed products by UPLC-MS. Hepatotoxic bioassay method cloud evaluate the toxicity of different Polygoni Multiflori Radix samples. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix, high pressure steamed three hours attenuated was better. Different processing methods have different effects on chemical constituents of Polygoni Multiflori Radix. Comparing with crude sample, the contents of gallic acid, 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside, emodin-8-O-beta glucoside and emodin were decreased in processed products with 3 kinds of different methods. The change trend of 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside content was similar with hepatotoxic potency. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix. Processing methods and time attenuated obvious impact on toxicity. Recommended further research on the attehuated standard control of Polygoni Multiflori Radix concocted.
Biological Assay ; Cell Line ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Fallopia multiflora ; chemistry ; toxicity ; Hepatocytes ; drug effects ; Humans ; Plant Roots ; chemistry ; toxicity

OBJECTIVETo develop a cellular assay based on heat shock signal pathway and secreted alkaline phosphatase (SEAP) reporter gene for investigating/predicting the early toxicity of heavy metals on HeLa cells in Chinese traditional medicine (TCM).

METHODThe pHSE-SEAP plasmid was transfected into HeLa cells to build a HSE-SEAP-HeLa cell model. For validation of the model, the transfected cells were treated by either heating at 42 degrees C for 1 h or incubated with 5 mol x L(-1) CdCl2 for 4 h. Then the cells were covered in complete DMEM culture medium for 48 h and the activity of SEAP (reflecting the cellular level of heat shock protein) in cultural supernatants was measured; meanwhile, cell viability was determined by MTT assays. In addition, the cells were treated by four mercury compounds, HgCl2, merthilate sodium, HgS and cinnabar at the sub-lethal concentrations (determined by MTT assays). Then the heat shock response was detected likewise.

RESULTSignificant level of secreted alkaline phosphatase (SEAP) was found in pHSE-SEAP transfected HeLa cells treated either by heating (42 degrees C) or incubating with CdCl2. The heat shock protein was induced by CdCl2 before decrease of cell viability was observed. All four mercury compounds induced heat shock response in both time and concentration-dependant manner. However, there were big differences among the mercury compounds, suggesting potential differences for early-stage toxicity in vivo.

CONCLUSIONThe pHSE-SEAP transfected HeLa cells respond effectively to heat shock and metal stresses, and therefore provide a practical and repeatable assay for investigating/predicting the early toxicity of heavy metals and mineral-containing drugs in TCM.

Alkaline Phosphatase ; biosynthesis ; genetics ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Genes, Reporter ; HeLa Cells ; Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Mercuric Chloride ; administration & dosage ; toxicity ; Mercury Compounds ; administration & dosage ; toxicity ; Metals, Heavy ; administration & dosage ; toxicity ; Plasmids ; Response Elements ; genetics ; Time ; Transfection