Actins ; metabolism ; Adult ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Calcium-Binding Proteins ; metabolism ; Cell Differentiation ; Desmin ; metabolism ; Diagnosis, Differential ; Female ; Fibroadenoma ; metabolism ; pathology ; surgery ; Hamartoma ; pathology ; Humans ; Hyperplasia ; Leiomyoma ; pathology ; Microfilament Proteins ; metabolism ; Muscle, Smooth ; pathology ; Phyllodes Tumor ; pathology

Xuefu Zhuyu decoction (XFZYD) is a famous traditional Chinese medicine (TCM) formula, is widely used in the treatment of cardiovascular and cerebrovascular diseases in China over one hundred years. But its effect on antioxidant and drug-metabolizing enzymes are unknown. This study was to observe the effects of Xuefu Zhuyu decoction (XFZYD) on the activities of antioxidant and drug metabolism enzymes (DMEs) in liver of rats. Male SD rats, treated with XFZYD at the dosage of 3.51, 7.02 and 14.04 g x kg(-1) per day for 15 days, serum were collected, tissue fluid, cytosols and microsomes isolated from liver tissues were prepared by centrifugation according to the standard procedure, the activities of antioxidant enzymes and drug-Metabolizing Enzymes were determined by UV-V is spectrophotometer. In serum, the activities of AST was not significantly affected by the treatment with XFZYD, at the high- est dose, the levels of ALT, Cr and BUN were significantly decreased (P < 0.05). GPX were significantly increased at the dose of 7.02, 14.04 g x kg(-1) (P < 0.05), CAT were significantly increased at the highest dose (P < 0.05). T-SOD was not significantly af- fected by this treatment. In the liver tissue, GPX was significantly increased at the dose of 3.51, 7.02 g x kg(-1) (P < 0.05), GST, CAT and T-SOD were not significantly affected following this treatment. In cytosols, GST was significantly increased at the dose of 3.51 g x kg(-1) (P < 0.05), T-SOD was remarkable induced at the dose of 3.51 and 7.02 g x kg(-1) (P < 0.05). In microsomes, XFZYD had no significant effect on Cytochromeb5, NADPH-Cytochrome P450 reductase, CYP3A, CYP2E1 and UGT, XFZYD significantly in- duced GST at the dose of 3.51 and 7.02 g x kg(-1) (P < 0.05), and the level of GSH were significantly increased by XFZYD at the dose of 3.51, 7.02 and 14.04 g kg(-1) (P < 0.05). These findings suggest XFZYD can induce the activities of GPX, CAT, SOD, GST and increase GSH level in liver of rats, which indicate XFZYD may have detoxification and antioxidant functions.
Animals ; Antioxidants ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Inactivation, Metabolic ; drug effects ; Liver ; drug effects ; enzymology ; Male ; Rats ; Rats, Sprague-Dawley

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

This paper introduced an experimental study of polylacticacid (PLA) nanoparticles of lipophilic anti-cancer herb drug using a precipitation method. Cucurbitacins (Cu) and Curcuminoids (Cur) were selected to be model drugs. They had similar solubility but their incorporation effects were significantly different: the average drug entrapment ratio, the average drug loading and the average drug recovery were 38.53%, 2.21% and 27.02% respectively; while those of Cur-PLA-NP were 94.36%, 14.35% and 91.23% respectively. To analyse the reason, drug incorporation process was investigated. By measuring solvent evaporation rate, ratio of drug PLA precipitates, drug distribution in system and entrapping ratio at different time of preparation, we found the difference of precipitation velocity of drug was the main reason. We also concluded that not all lipophilic drug can be well entrapped into PLA nanoparticle by nanoprecipitation method. The drug incorporation depended on the interations among drug, PLA and organic solvents, in addition to the solubility of the drug.
Antineoplastic Agents, Phytogenic ; chemistry ; Chemical Precipitation ; Cucurbitacins ; chemistry ; Curcumin ; chemistry ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Lactic Acid ; chemistry ; Nanoparticles ; Nanotechnology ; methods ; Particle Size ; Polyesters ; Polymers ; chemistry

BackgroundHuman retinal gliocytes play an important role in proliferative diseases,which are the basis of in vitro studies.Researchers have cultured human retinal gliocytes in the past.In our study,we found that the cells we cultured presented a unique shape different from those by other researchers.ObjectiveThis study was to design to produce a new culture and purification method for retinal gliocyte in vitro.Methods Retinal tissue was isolated from human eyeballs and digested using the two-step digestion method (2％ pancreatin and 0.133％collagenase Ⅵ) to harvest the retinal glio cytes.The cells were collected and cultured in endothelial cell-targeted nutrient culture containing 10％ fetal calf serum and supplemented with β-endothelial cell growth factor (ECGF) and sodium heparin,and the culture dishes were coated with fibronectin(FN) to promote the attachment of retinal gliocyte.During the culturing process,the gliocytes were identified by the observation of morphological characteristic and regular histological examination.The identification of the cells also was performed by immunochemistry targeting glial fibrillary acidic protein (GFAP),Vimentin,neuron specific enolase ( NSE ),S-100,CD34,and Ⅷ factor.Results Retinal gliocytes were isolated successfully from the human retina by the two-step digestion method.Primary cultured cells attached after 72 hours and achieved confluency between day 9 and 10 that were aligned petaliform in shape.Regular histological examination after H&E staining showed blue cell nuclei and light red cytoplasm.The target cells presented with strong responses for GFAP and Vimentin and no response for NSE,S-100,CD34 and Ⅷ factor.ConclusionsLarge amount of purified human retinal gliocytes can be obtained by two-step digestion and cultured in endothelial cells-targeted culture medium supplemented with β-ECGF and sodium heparin in plates coated with FN.The cultured cells expressed markers for retinal gliocytes.However,specific features of these cells remain to be further elucidated.

By setting up the organic additive model of chemical fingerprints of TCM-compound, the quantified fingerprint method had been established to solve the qualitative and quantitative analyses problems for both the fingerprint attribution ratio and process recovery of medicinal effective components in TCM-compound prescription. The method firstly performs the qualitative analyses of the attribution ratios, and then the quantitative analyses, which can successfully disclose the results of attribution ratio and determine the process recovery of the medicinal effective components for TCM-compound prescription. Three optional methods were represented to assess the amount and distribution proportion of chemical compositions for single crude drug to compound prescription. In terms of components absorbed ultraviolet light, S5 (Radix Scutellariae) was assessed to be the most important crude drug containing much more effective components, and S7 (Radix Gentianae), S4 (Flos Lonicerae Japonica), S8 (Rhizome Anemarrhena) and S9 (Fructus Gardeniae) were second important crude drugs. The results showed lower process recovery of the medicinal effective components for eight batches of marketed preparations. Above all, the quantified fingerprint method can objectively and accurately reflect how high is the contribution of a single crude drug to the compound prescription, and quantitatively evaluate the process recovery of medicinal effectiveness components.
Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; analysis ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the mechanism of lemon peel essential oil (LPE) on the cariogenicity of Streptococcus sobrinus (Ss).

METHODSLPE was extracted by the authors, and the minimum inhibition concentration (MIC) was measured by disc diffusion method. The LPE was used as the experimental group with concentrations ranging from 2.250 g/L to 0.281 g/L prepared with trypticase peptone yeast (TPY) culture medium, and TPY culture medium was used as the control group. Ss at the concentration of 10(8) CFU/ml was added to each group, and cultured for 6, 18, 24, 48 hours. Neson-Somogyi method was used to measure the content of reducing sugar, and glucosyltransferase (GTF) activity. The activity of lactate dehydrogenase (LDH) was measured by lactic acid and pyruvic acid continuous monitoring method. The content of water insoluble glucan (WIG) was measured by anthrone method, and the pH value of the culture solution was detected. The value of pH before the experiment and the time difference was alculated as ΔpH.

RESULTSAt the same time point, the activity of GTF and LDH and the concentration of WIG and the value ΔpH decreased gradually with the increase of concentration of LPE. There were significant differences between each experimental group and control group (P < 0.01). The control group had the maximum value, GTF: (6.71 ± 0.61) mIU, LDH: (135.8 ± 1.7) U/L, WIG: (47.15 ± 5.12) mg/L, ΔpH: (2.67 ± 0.01). The highest drug concentration group had the minimum value: GTF: (0.39 ± 0.07) mIU, LDH: (95.0 ± 5.4) U/L, WIG: (2.44 ± 0.38) mg/L, ΔpH: (0.61 ± 0.01).

CONCLUSIONSThe LPE below the MIC could still inhibit the GTF, LDH activity and lead to the decrease of WIG and the acid production.

Dose-Response Relationship, Drug ; Glucans ; biosynthesis ; Glucosyltransferases ; antagonists & inhibitors ; metabolism ; Lactate Dehydrogenases ; antagonists & inhibitors ; metabolism ; Microbial Sensitivity Tests ; Oils, Volatile ; pharmacology ; Plant Oils ; pharmacology ; Streptococcus sobrinus ; drug effects ; metabolism

OBJECTIVETo obtain the optimized extraction technology of Paeonia suffruticosa by comparing several extraction method.

METHODExtract P. suffruticosa by ethanol circumfluence, distillation-decoction, CO2-SFE and traditional decoction, and analyse the results according to the total extraction rate, extraction rate of paeono, extraction of other ingredients and production feasibility.

RESULTTotal extraction rates of which are 12.66%, 13.51%, 7.28%, 7.56% respectively; extraction rates of paeonol are 2.45%, 2.26%, 0.31%, 1.15% in turn; Phenolic glycosides can be extracted by ethanol circumfluence, distillation-decoction, traditional decoction, but not by CO2-SFE.

CONCLUSIONDistillation-decoction is the most proper extraction technology of P. suffuticosa at present.

Acetophenones ; isolation & purification ; Drugs, Chinese Herbal ; isolation & purification ; Paeonia ; chemistry ; classification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods

This paper introduced an experimental study of the preparation of polylacticacid (PLA) nanoparticles of cucurbitacin (CuC) using a precipitation method. The residual acetone, ratio of CuC PLA precipitates, and the relationships between the ratios of two precipitates and drug incorporation rates were measured. It appeared that the nanoparticles with 60% of PLA incorporated with 5.5% of CuC were formed when acetone was injected into the aqueous phase. As the acetone gradually evaporated, drug incorporation/encapsulation continued, with most of CuC (about 70%) formed new crystalline cores and suspended in the form of microcrystals in the medium, resulting a suspension containing both nanoparticles and microcrystals. We also concluded that this system may not necessarily be suitable for all lipophilic drugs to be prepared to PLA nanoparticles with good incorporation rate. The drug incorporation depended on the interactions among drug, PLA, and organic solvents, in addition to the solubility of the drug.
Antineoplastic Agents, Phytogenic ; administration & dosage ; Chemical Precipitation ; Cucurbitaceae ; chemistry ; Cucurbitacins ; Delayed-Action Preparations ; Drug Compounding ; methods ; Drug Delivery Systems ; Lactic Acid ; Microspheres ; Nanotechnology ; Particle Size ; Plants, Medicinal ; chemistry ; Polyesters ; Polymers ; Triterpenes ; administration & dosage ; isolation & purification

Adenocarcinoma ; metabolism ; pathology ; surgery ; Cardia ; Esophageal Neoplasms ; metabolism ; pathology ; surgery ; Esophagogastric Junction ; metabolism ; pathology ; surgery ; Humans ; Neoplasm Staging ; Receptor, ErbB-2 ; metabolism ; Sirtuin 1 ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; Survival Rate