0.05).At different time points after ischemia-reperfusion,the expression of cytochrome C and activation of caspase-3 were lower in the transgen mice than that in the wild type rats.Conclusions Under standard condition,overexpression of bcl-xl could significantly reduce the infarct area and improve neurological function in transgene mice than those in the wild type rats.The effect of overexpression of bcl-xl might be realized through inhibiting the apoptosis of neuron,and the mechanism might be that the overexpression of bcl-xl inhibit the release of cytochrome C and the activation of caspase-3.

The endophytic microorganisms found widely in many kinds of plants mediate various effects to theirs hosts. In this study, seven different dominant endophytes (SDE01 to 07) isolated from a Hy-peraccumulator-Solanum nigrum L. were resistant to Cd2+, and the strain SDE06 survived even in the medium containing 80 mg/L of Cd2+. Bacteria strain SDE06 was identified as Bacillus sp.. The removal of Cd2+ of SDE06 in different conditions were studied. Under the optimal conditions, the incubating time was 36 h, the solution pH 6.0, the temperature was 37?C and the Cd2+ concentration of medium was 20 mg/L, the highest removal rate was up to 80.2% at this condition.

In the present study, a new compound named 17-(6-cinnamamido-hexylamino-)-17-demethoxygeldanamycin (CDG) was obtained by introducing the cinnamic acid (CA) group into the 17-site of geldanamycin (GDM). The anti-cancer effects of CDG in vitro and in vivo were evaluated. MTT assay was used to examine the inhibitory effect of CDG on the proliferation of MCF-7, HepG2, H460 and SW1990 cells. Immunofluorescent staining flow cytometry combined with Annexin V-FITC/PI staining were used to detect apoptotic cells. Transwell assay was used to analyze the effect of CDG on cell invasion and migration ability. Western blotting was used to detect the expression levels of RAF-1, EGFR, AKT, CDK4 and HER-2 of MCF-7, HepG2 and H460 cells. The toxicities of CDG and GDM were evaluated in mice. Using the subcutaneously transplanted MCF-7 xenograft in nude mice, inhibitory effect was evaluated in vivo. The results showed that CDG inhibited the proliferation of cancer cells (IC50: 13.6-67.4 microg.mL-1). After exposure to CDG for 48 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation or shrunken nucleus. The rates of apoptosis of MCF-7, HepG2, H460 and SW1990 cells incubated with 10 microg.mL-1 CDG were 23.16%, 27.55%, 22.21%, 20.47%, respectively. A dose-dependent reduction of migration of four cell lines was found after exposure to CDG. The decreased levels of RAF-1, EGFR, AKT, CDK4 and HER-2 showed that CDG possessed HSP90 inhibitory effect. The result of animal toxicity test on the mice suggested that CDG had lower toxicity than GDM. Meanwhile, CDG inhibited the growth of MCF-7 xenografts of athymic mice.
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Benzoquinones ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase 4 ; metabolism ; Female ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Lactams, Macrocyclic ; chemical synthesis ; chemistry ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Proto-Oncogene Proteins A-raf ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; metabolism ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

From an aqueous extract of Lonicera japonica flower buds, sixteen compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, MCI gel, silica gel, and sephadex LH-20 and reversed-phase HPLC. Their structures were elucidated by spectroscopic data analysis as 6'-O-acetylvogeloside (1), 6'-O-acetylsecoxyloganin (2), dichlorogelignate (3), guanosinyl-(3' --> 5')-adenosine monophosphate(GpA,4) , 5'-O-methyladenosine (5), 2'-O-methyladenosine (6), adenosine (7), syringin (8), methyl 4-O-β-D-glucopyranosyl caffeate (9), (-)-dihydrophaseic acid 4'-O-β-D-glucopyranoside (10), ketologanin (11), 7α-morroniside (12), 7β-morroniside (13), kingiside (14), cryptochlorogenic acid methyl ester (15), and 6-hydroxymethyl-3-pyridinol (16). All the compounds were obtained from this plant for the first time, compounds 1 and 2 are new compounds, 3 and 5 are new natural products, and 4 is the first example of dinucleoside monophosphate isolated from a plant extract.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flowers ; chemistry ; Lonicera ; chemistry ; Mass Spectrometry ; Molecular Structure

AIMTo investigate the inhibitory activity of salvianolic acid A (SAA) on nucleoside transport in cancer cells and its antitumor effect.

METHODS[3H] thymidine and [3H] uridine transport assays were used to determine the inhibitory activity on nucleoside transport in Ehrlich carcinoma cells. The cytotoxicity to cultured cancer cells was examined with clonogenic assay. The antitumor effect in vivo was evaluated with transplantable tumor model in mice.

RESULTSSAA was shown to inhibit thymidine and uridine transport in Ehrlich carcinoma cells with IC50 values of 18.1 and 17.1 micromol x L(-1), respectively. By clonogenic assay, the IC50 of SAA for KB cells was 44.7 micromol x L(-1). SAA markedly potentiated the cytotoxicity of 5-FU and mitomycin C in KB cells as well as the cytotoxicity of MTX in human hepatoma BEL-7402 cells. For in vivo experiment, sarcoma 180 cells were transplanted sc in mice and tested drugs were administered ip. When administered separately, SAA at 200 mg x kg(-1) and 5-FU at 10 mg x kg(-1) inhibited tumor growth by 41% and 27%, respectively. Combination of the two drugs inhibited tumor growth by 63% (CDI = 0.86).

CONCLUSIONSAA is active in blocking nucleoside transport in cancer cells and potentiates the cytotoxicity of chemotherapeutic drugs. As an agent showing moderate antitumor effect in vivo, SAA might be useful in combination cancer therapy.

Animals ; Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Biological Transport ; Caffeic Acids ; isolation & purification ; pharmacology ; Drug Synergism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Fluorouracil ; pharmacology ; Humans ; KB Cells ; Lactates ; isolation & purification ; pharmacology ; Male ; Methotrexate ; pharmacology ; Mice ; Mitomycin ; pharmacology ; Neoplasm Transplantation ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry ; Sarcoma 180 ; metabolism ; pathology ; Tumor Cells, Cultured

To investigate the effect of lidamycin (LDM) on human gastric carcinoma BGC823 cells and xenograft growth in nude mice, MTT (methyl thiazolyl tetrazolium) assay was used to determine the inhibition of BGC823 cell proliferation by LDM. Induction of apoptosis was studied by flow cytometry and TUNEL assay. The expression of VEGF was detected by Western blotting analysis. Athymic nude mice were used to determine in vivo antitumor activity. Proliferation inhibition and apoptosis induction were studied in lidamycin-treated cells. The expression of VEGF in BGC823 cells decreased in a dose-dependent manner. LDM at 0.02 mg x kg(-1) and 0.04 mg x kg(-1) suppressed the growth of BGC823 xenografts in nude mice by 57% and 72%, respectively. LDM potently induces apoptosis in human gastric carcinoma BGC823 cells and inhibits xenograft growth.
Aminoglycosides ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Enediynes ; pharmacology ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Random Allocation ; Stomach Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays

Recent studies have shown that epidermal growth factor receptor (EGFR) is an important target for cancer therapy. The present study prepared single chain Fv (scFv) directed against EGFR. Balb/c mice were immunized by human carcinoma A431 cells, and total RNA of the splenic cells was extracted. VH and VL gene fragments were amplified by RT-PCR and further joined into scFv gene with a linker, then scFv gene fragments were ligated into the phagemid vector pCANTAB 5E. The phagemid containing scFv were transformed into electro-competent E. coli TG1 cells. The recombinant phage antibody library was constructed through rescuing the transformed cells with help phage M13K07. The specified recombinant phages were enriched through 5 rounds of affinity panning and the anti-EGFR phage scFv clones were screened and identified with ELISA. A total of 48 clones from the library were selected randomly and 45 clones were identified positive. After infecting E. coli HB2151 cells with one positive clone, soluble recombinant antibodies about 27 kD were produced and located in the periplasm and the supernatant. The result of sequencing showed that the scFv gene was 768 bp, which encoded 256 amino acid residues. VH and VL including 3 CDRs and 4 FRs, respectively, were all homologous to mouse Ig. The soluble scFv showed the specific binding activity to purified EGFR and EGFR located in carcinoma cell membrane. The successful preparation of anti-EGFR scFv will provide an EGFR targeted molecule for the development of antibody-based drugs and biological therapy of cancer.
Animals ; Antibodies, Monoclonal ; Cell Line, Tumor ; Humans ; Immunoglobulin Light Chains ; genetics ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Peptide Library ; Receptor, Epidermal Growth Factor ; genetics ; immunology ; Single-Chain Antibodies ; genetics ; immunology

OBJECTIVETo understand clinical application of syndrome differentiation and treatment of meridians in acupuncture and moxibustion.

METHODSCollect and collate the literature about syndrome differentiation and treatment of meridians in Chinese Acupuncture & Moxibustion, Acupuncture Research, Shanghai Journal of Acupuncture & Moxibustion (Clinic edition B in 2003) and Clinical Journal of Acupuncture and Moxibustion from 2001 to 2003.

RESULTSLess literature about syndrome differentiation and treatment of meridians are involved, only accounting for 5.69% (163/2 864); and the literature about both treatment based on meridians and treatment based on syndrome differentiation of meridians only accounted for 1.22% (35/2 864).

CONCLUSIONAt present, clinically acupuncture and moxibustion do not play attention to treatment based on syndrome differentiation of meridians.

Acupuncture Points ; Acupuncture Therapy ; China ; Humans ; Meridians ; Moxibustion

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

OBJECTIVETo observe the clinical effect of acupoint injection with small dose of fentanyl-droperido mixed liquor at different time stages on labor analgesia, and explore its function mechanism.

METHODSOne hundred and fifty cases of full-term primiparas who were intended to take vaginal delivery were randomly divided into 3 groups, 50 cases in each one. The acupoint injection with fentanyl-droperido mixed liquor at different time stages was applied in the group I, where Shenshu (BL 23) was selected in active phase and Ciliao (BL 32) was selected in the 2nd stage of labor. The subcutaneous injection with fentanyl-droperido mixed liquor was adopted in group II. The group III, which was considered as a control group, was treated with subcutaneous injection of 0.9% NaCl at the same time stage as group I and II. The blood pressure, VAS score and level of norepinephrine and adrenaline were observed at different time stages.

RESULTSCompared before the injection, the blood pressure of group III in the active phase and 2nd stage of labor was significantly increased (P < 0.05), and that in group II and III was obviously higher than that in the group I (P < 0.05). The VAS score of group I in the active phase and 2nd stage of labor was significantly lower than that in the group II and III (P < 0.01). After the injection, the level of NE and E was significantly decreased (P < 0.05), which was lower than that in the group II and III (P < 0.05).

CONCLUSIONThe acupoint injection with small dose of fentanyl-droperido mixed liquor at different time stages has positive effect on labor analgesia, and it could significantly relieve stress reaction of parturient during the labor. Its mechanism could be related to the reduction of stress hormone in parturient.

Acupuncture Points ; Adult ; Analgesia, Obstetrical ; Analgesics ; administration & dosage ; Delivery, Obstetric ; Droperidol ; administration & dosage ; Female ; Fentanyl ; administration & dosage ; Hormones ; metabolism ; Humans ; Labor Pain ; drug therapy ; Labor, Obstetric ; metabolism ; Pregnancy ; Young Adult