OBJECTIVETo extract RNA from Pinellia ternata and lay a foundation for studying the formation mechanism of P. ternata.

METHODBy modifying the method recommended by Guanidinium for extracting total RNA from plant tissues rich in phenolic and polysaccharidic compounds, a simple and convenient method for extraction of total RNA from the tubers, stems and leaves of P. ternate containing abundant polyphenols and polysaccharides was established. High concentrated p-mercaptoethanol was added in the RNA extracted buffer to remove polyphenols, phenol and chloroform were used to eliminate proteins, and isopropanol and sodium acetate were used to precipitate polysaccharides.

RESULTThe A260/A230 value of RNA extracted with improved method were all over 2.0 and the values of A260/A280 were between 1.7 and 2.0. The electrophoresis bands were cleared on agarosegel and integrity of RNA was good.

CONCLUSIONThe results showed that RNA obtained from the tubers, stems and leaves of P. ternate with this method had high purity and quality and could be used in molecular biological research, as DDRT-PCR and reverse Northern blotting analysis directly. This method is simple, economic, stable performance, and has a good repeatability as well as is suitable for extracting total RNA of medicinal plants with high concentrations of phenolics and polysaccharides.

Blotting, Northern ; Pinellia ; genetics ; Plant Leaves ; genetics ; Plant Stems ; genetics ; Plant Tubers ; genetics ; Polymerase Chain Reaction ; RNA, Plant ; isolation & purification

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects

Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.

OBJECTIVETo observe the anti-viral therapy effect on HBV reactivation in malignant tumor patients and hepatitis B virus carriers after their cancer chemotherapy.

METHODSThirteen cancer patients but also chronic hepatitis B virus carriers were enrolled in this study. They were randomly put into two groups. Eight patients were put in the therapeutic group. They all had abnormal liver functions induced by the reactivation of HBV after their cancer chemotherapy. Then they were treated with lamivudine. The other 5 cases were treated with lamivudine before their cancer chemotherapy when their serum HBV DNA levels were less than 10(3) copies/ml (preventive therapeutic group). The two groups were followed-up with liver function tests and serum HBV DNA level measurements.

RESULTSAmong the 8 cases of the therapeutic group, 5 cases died of liver failure; cancer chemotherapy was postponed or even terminated in 3 patients due to liver function abnormality and anti-virus treatment was started. In the preventive therapy group, no HBV reactivation was observed in any of the 5 cases.

CONCLUSIONFor HBV carrier cancer patients, an anti-viral therapy before their cancer chemotherapy seems to be very important.

Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; Antiviral Agents ; therapeutic use ; Carrier State ; virology ; Female ; Hepatitis B ; virology ; Hepatitis B virus ; drug effects ; Humans ; Lamivudine ; therapeutic use ; Male ; Neoplasms ; drug therapy ; virology ; Virus Activation ; drug effects

Objective To explore the effect of nursing leadership on health education in patients after stroke.Methods Inpatients after first stroke received health education provided by nurses.When the patients went back to out-patient department after discharge, 64 of them were selected as subjects.The Stroke Knowledge Questionnaire (SKQ) and Health Promoting Lifestyle Profile Ⅱ (HPLP Ⅱ) were used.Results Except "Spiritual Growth", the scores of SKQ and other subscales of HPLP Ⅱ were higher than those before stroke.But the standard score of SKQ after stoke was only (67.14±17.18).Conclusions Our health education can effectively enhance the health knowledge and behavior in patients after stroke, but it is not satisfied, it is important to enhance nurses' leadership to improve our health education.

OBJECTIVETo study the chemical constituents from the roots of Craibiodendron henryi.

METHODVarious chromatographic techniques were used to isolate and purify the constituents. The structures were elucidated by chemical evidence and spectral methods.

RESULTTwelve compounds were isolated from the ethyl acetate soluble fraction of the 95% ethanolic extract and their struc- tures were elucidated as quercetin (1), quercetin-3-O-rhamnicoside (2), quercetin-3-O-arabinofuranoside (3), (-)-epicatechin (4), proanthocyanidin A-2 (5), procyanidin B-2 (6), (-)-isolariciresinol-2a-O-beta-D-xylopyranoside (7), lyoniside (8), sitoster-yl-3beta-glucoside-6'-O-palmitate (9), beta-sitosterol (10), daucosterol (11) and octacosanoic acid (12).

CONCLUSIONCompounds 1-12 were isolated from this plant for the first time.

Biflavonoids ; chemistry ; isolation & purification ; Catechin ; chemistry ; isolation & purification ; Ericaceae ; chemistry ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Proanthocyanidins ; chemistry ; isolation & purification ; Quercetin ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification

OBJECTIVETo study the chemical constituents of Cynanchum forrestii.

METHODChromatographic techniques were applied to isolated chemical constituents. The structures were identified on the basis of physico-chemical constants and spectroscopic data.

RESULTEight compounds were isolated from the 95% ethanol extract of the roots of C. forrestii and elucidated as ( + ) -5'-methoxyisolariciresinol 3a-O-beta-D-glucopyranoside (1), hexahydroxycholest-7-en-6-one (2), tylophorinidine (3), sucrose (4), palmitic acid (5), beta-sitosterol (6), daucosterol (7), nonanedioic acid (8).

CONCLUSIONCompounds 1-3 from this genus, and compounds 4-8 from the plant were obtained for the first time.

Alkaloids ; chemistry ; isolation & purification ; Cholestenones ; chemistry ; isolation & purification ; Cynanchum ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Isoquinolines ; chemistry ; isolation & purification ; Palmitic Acid ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVEIn this study, orthogonal design was used to optinize DDRT-PCR amplification system on Pinellia ternata microtubers in vitro in five factors four levels respectively.

METHODP. ternata stems and microtubers in vitro were selected as explants. The effects of five kinds of factors were studied by orthogonal design method including emplate cDNA, Mg2+, dNTPs, primers and Taq DNA polymerase, and in order to establish the optimum DDRT-PCR system of P. ternata microtubers in vitro.

RESULT AND CONCLUSIONA satisfactory DDRT-PCR technique system for P. ternata microtubers in vitro with desirable repeatability and polymorphic bands was established. In a total volume of 20 microL DDRT-PCR system, it contained 10 x buffer, 150 micromol L(-1) dNTPs, 2 micromol L(-1) anchor primer, 1 micromol L(-1) arbitrary primer, 2.5 mmol L(-1) Mg2+, 0.6 U Taq DNA polymerase and 2.5 microg template cDNA. The effect of the five factors was in sequence of Taq DNA polymerase > template cDNA > dNTPs > Mg2+ > Primers. The optimum DDRT-PCR system will provide scientific reference basis for studying effecting character of P. ternata microtubers associated with genes expression.

DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Electrophoresis, Polyacrylamide Gel ; Pinellia ; genetics ; Plant Tubers ; genetics ; Polymerase Chain Reaction ; methods ; Silver Staining ; Taq Polymerase ; genetics

OBJECTIVETo investigate the effects of beta1-integrin, fibronectin (FN) and laminin (LN) on the invasive behavior of human gliomas.

METHODSFunctional impacts of beta1-integrin, fibronectin and laminin on cell adhesion, migration and metastasis of U251 malignant glioblastoma cells were investigated by in vitro adhesion, migration and invasion assays. The amount and distributions of cellular microfilaments and pseudopodia were studied by fluorescent cytochemistry, confocal laser scanning microscope and scanning electron microscope. Lastly, beta1-integrin, fibronectin and laminin were investigated for their roles in cellular microfilament skeleton.

RESULTS(1) Fibronectin did not affect cell adhesion of U251MG cells, but anti-beta1 integrin antibodies inhibited cell adhesion (P < 0.01); Laminin stimulated cell adhesion of U251MG cells (P < 0.01) but anti-beta1 integrin antibodies had little effect on the laminin-mediated cell adhesion. (2) The migration of U251MG cells on dishes coated with FN was inhibited by anti-beta1 integrin antibodies (P < 0.05). (3) F-actins formed strong and dense stress fibers in U251MG cells on dishes coated with FN and LN. Anti-beta1 integrin antibodies disrupted the microfilament network and F-actin aggregation. (4) FN and LN increased the number of pseudopodia on cell surface, whereas anti-beta1 integrin antibodies reversed this function. (5) FN and anti-beta1 integrin antibodies had little effects on the invasive ability of U251MG cells in vitro. The invasion was increased by LN, but inhibited by anti-beta1 integrin antibodies.

CONCLUSIONS(1) The interaction between beta1-integrin, FN may stimulate U251MG cell migration via changing the structures of microfilament skeleton and the number of pseudopodia. (2) beta1-integrin may play a role in the LN-mediated in vitro invasion of U251MG cells.

Cell Adhesion ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Fibronectins ; pharmacology ; Glial Fibrillary Acidic Protein ; analysis ; Glioma ; metabolism ; pathology ; ultrastructure ; Humans ; Immunohistochemistry ; Integrin beta1 ; pharmacology ; Laminin ; pharmacology ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Neoplasm Invasiveness

OBJECTIVETo establish and characterize imatinib-resistant gastrointestinal stromal tumor (GIST) xenografts. Further provided an ideal experimental platform through the imatinib-resistant GIST xenografts to investigate the mechanism of resistance to imatinib.

METHODSImatinib-resistant GIST cells were injected under the skin of athymic nude mice to establish animal models of human imatinib-resistant GIST. The molecular and histopathologic features of GIST xenografts were also analysed and compared with their counterpart of cell lines.

RESULTSThe xenograft tumor models had been established by subcutaneously injection of GIST cells into nude mice. Immunohistochemistry results showed CD117 expression was positive in GIST-PR2 xenograft tumor, but negative in GIST-R. In GIST-PR1, tumor areas showing rhabdomyoblastic differentiation were presented next to areas with classic GIST morphology. The rhabdomyoblastic component demonstrated consistently positivity for desmin and myogenin, whereas CD117 was completely negative. The mutation profiles of these xenograft tumors were the same as their counterpart of cell lines.

CONCLUSIONSHuman GIST xenografts with mutation in c-kit have been established from imatinib-resistant GIST lines. Those models will enable further studies on mechanisms of resistance, combination therapies and allow testing of novel targeted therapies.

Animals ; Antineoplastic Agents ; pharmacology ; Benzamides ; Cell Differentiation ; Cell Line, Tumor ; Desmin ; metabolism ; Drug Resistance, Neoplasm ; Female ; Gastrointestinal Neoplasms ; genetics ; metabolism ; pathology ; Gastrointestinal Stromal Tumors ; genetics ; metabolism ; pathology ; Humans ; Imatinib Mesylate ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mutation ; Myogenin ; metabolism ; Piperazines ; pharmacology ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Pyrimidines ; pharmacology ; Rhabdomyosarcoma ; metabolism ; pathology ; Xenograft Model Antitumor Assays