Objective To investigate the therapeutic effects of microwave ablation combined with dendritic cell (DC) tumor vaccine on a murine breast cancer model and to explore its immunological mechanism.Methods Forty-eight Balb/c mice were used to establish models of mouse mammary carcinoma,and then randomly divided into a sham-operation group,a microwave ablation treated group,a DC tumor vaccinated group and a microwave ablation plus DC tumor vaccinated group.Microwave ablation was conducted with the microwave ablation group on day 14 after establishing the model.DC tumor vaccination was performed on day 7 in the vaccinated group.In the microwave ablation plus DC tumor vaccinated group,DC tumor vaccination and microwave ablation were consecutively performed on days 7 and 14,respectively.Tumor growth,lung metastasis and survival were observed.The proportion of CD3+ and CD4+ T cells in splenocytes were analyzed by fluorescence Activated Cell Sorter (FACS) on the 14th day after microwave ablation.Serum levels of interferon-γ (IFN-γ) and interleukin-4 (IL-4) were also detected using ELISA.Results The survival time (56.4 ± 4.3 days),tumor size (67.3 ± 6.1 mm2) and lung metastasis index (0.6 ± 0.4) in the microwave ablation plus DC tumor vaccinated group were all significantly lower than the corresponding averages in the microwave ablation and DC tumor vaccinated group.The proportions of splenic CD3 + T cells (27.8 ±10.3 ％) and CD4+ T cells (16.7 ± 8.3 ％) in the microwave ablation plus DC tumor vaccinated group were significantly larger than in the control group.Average serum levels of IFN-γ (41.8 ± 22.9 pg/ml) in the microwave ablation plus DC tumor vaccinated group were also significantly larger than in the control group,while the average serum level of IL-4 (15.4± 6.4 pg/ml) was significantly lower.Conclusions Microwave ablation and DC tumor vaccination together could have synergistic effects in treating experimental breast cancer,which might be closely related to the enhanced immune response.

Traditional medicine (herb medicine) began to prevail again over last two decades, and it is about 70% of the world population taking herb medicine as supplement or alternative medicine according to a recent survey. The consumption of herb medicine increased exponentially in Canada, Australia and Europe during last 10 years. Since concomitant administration of herbal and western medicine has become a trend, it requires paying close attention to the problem. Herb-drug interactions have been extensively investigated worldwide, and there is an increasing concern about the clinical herb-drug interaction. In this review we introduced the current progress in the herb-drug interactions including evidence-based clinical studies and establishment of levels of evidence for herb-drug interaction; and in the related mechanisms including the induction and inhibition of metabolic enzymes, inhibition and induction of transport and efflux proteins, alteration of gastrointestinal functions, and alteration in renal elimination. We also analyzed both the achievements and the challenges faced in the concomitant administration of traditional Chinese medicine and western medicine.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Biological Transport ; drug effects ; Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; pharmacology ; Evidence-Based Medicine ; methods ; Gastrointestinal Tract ; drug effects ; Herb-Drug Interactions ; Humans ; Kidney ; drug effects ; Medicine, Chinese Traditional ; Pharmacokinetics ; Phytotherapy ; Plants, Medicinal ; chemistry

Objective To clne huma PAIl gene and prepare its monoclonal antibodies(McAbs) for determination of its expression in breast cancer cells.Methods Human PAI1 gene eDNA was amplified by RT-PCR from human breast cancer cell line MDA231 and inserted into the prokaryotic expression vector, which expressed fusion protein of MS2-PAI1 in E.coli.Fusion protein of MS2-PAI1 was purified and used for immunizing BALB/C mouse.Traditional hybridoma technology was used to produce hybridoma cells for preparation of monoclonal antibodies.Western blot and immunohistochemistry were used to detect PAI1 expression in breast cancer.Results The 1209 bp full PAI1 gene was cloned.The two hybridoma cell lines that secreted specific monoclonal antibodies against human PAI1 were identified by ELISA.The immunoglobulin subclasses of the McAbs were IgG1.The McAbs can specifically recognize PAI1 but not other proteins.Western blot showed that the McAbs against PAI1 can specifically react with MS2-PAI1 fusion protein and endogenous proteins in cells.The positive reaction was found in breast cancer cell line MDA231 and breast cancer tissues by immunochemical staining.Conclusions The McAbs against human PAI1 are successfully prepared by hybridoma technology with MS2-PAI1 fusion protein expressed in E.coll.It has been shown that PAI1 can be expressed in MDA231 and breast cancer tissues.The McAbs against PAI1 could be a useful tool for the further study of the human PAI1 functions and detection of clinical tumor samples.

To study the anti-tumor metastatic constituents in Rhodiola wallichiana (HK) S H Fu var Cholaensis (Praeg) S H Fu, chemical constituents were isolated and purified by repeated column chromatography (silica gel, Toyopearl HW-40C and preparative HPLC). Their structures were elucidated on the basis of spectral data analysis. The anti-tumor metastasis assay was applied to evaluate the activities of the isolated compounds. Ten compounds (1-10) were isolated and their structures were identified by comparison of their spectral data with literature as follows: syringic acid (1), salidroside (2), tyrosol (3), scaphopetalone (4), berchemol (5), 2,6-dimethoxyacetophenone (6), rhobupcyanoside A (7), miyaginin (8), chavicol-4-O-β-D-apiofuranosyl-(1 --> 6)-O-β-D-glucopyranoside (9), eugenyol-O-β-D-apiofuranosyl-(1 --> 6)-O-β-D-glucopyranoside (10). Compounds 4-6 and 8-10, were isolated from this genus for the first time, while compound 7 was isolated from this plant for the first time. Compounds 2, 6-8 showed positive anti-tumor metastatic activities, and compounds 2 and 8 showed significant anti-tumor metastatic activities.
Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Humans ; Neoplasm Metastasis ; prevention & control ; Rhodiola ; chemistry

In order to investigate whether cytokine-induced killer (CIK) cells can induce apoptosis of bcr-abl(+) K562 cells, apoptosis of K562 cells and CEM cells induced by CIK cells, etoposide or camptothecin was detected with flow cytometry DNA assay. RT-PCR showed that K562 cells expressed the bcr-abl fusion gene, K 562 cells, K562 cells/etoposide or K562 cells/camptothecin groups showed no sub-G(1) peak. K562 cells/CIK cells group showed sub-G(1) peak (38.1%). CEM cells showed no sub-G(1) peak. CEM cells/camptothecin or CEM cells/etoposide groups showed sub-G(1) peak (23.5% or 32.3% respectively). CEM cells/CIK cells group showed sub-G(1) peak (45.4%). Etoposide or camptothecin did not induce apoptosis of K562 cells. CIK cells induce apoptosis of K562 cells. Bcr-abl fusion gene prevented apoptosis induced by etoposide or camptothecin, but did not prevent apoptosis induced by CIK cells. This property may support the observed adoptive immunologic effect of allogeneic bone marrow transplantation and donor lymphocyte transfusions of CML case relapsing after allogeneic bone marrow transplantation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; immunology ; Camptothecin ; pharmacology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Etoposide ; pharmacology ; Flow Cytometry ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; drug effects ; immunology ; metabolism ; Killer Cells, Lymphokine-Activated ; cytology ; immunology

OBJECTIVETo explore the clinical effects of the manipulation reduction combined with small splint fixation for the treatment of fresh closed fracture of radius for shorten hospital stays and reduce medical cost.

METHODSFrom July 2007 to December 2009, 200 patients (ranged the age from 40 to 80 years) with distal radius comminute fracture were treated and divided into CP group (including 21 males and 79 females, with a mean age of (62.98 +/- 0.85) years), and control group (including 20 males and 80 females, with a mean age of (63.19 +/- 0.88) years). All patients were treated manipulation reduction combined with small-splint fixation, control group removed small-splint 30 days after treatment, CP group removed 25 days after treatment. Two groups were checked by X-ray and took traditional chinese medicine (taking Yuanhu tablets, Chuangshangning tablets on the early stage; Guixiangzhenggu pill was taken on the middle stage; Shuanglongjie gu pill on the late stage), functional exercise was guided after removing of small splint. The condition of reduction and position of bone were evaluated and Gartland-Werlley scale was used to evaluate the function of wrist joint.

RESULTSTreatment time in CP group was decreased from (30.08 +/- 3.06) to (25.06 +/- 1.07) days; treatment cost in CP group was decreased from (2 100.00 +/- 332.12) to (1 644.00 +/- 125.20) Yuan. There was no significant difference in reduction and function recover of wrist joint between two groups. The results showed the effects of TCM clinic can be promised.

CONCLUSIONClinical pathway for outpatient can promote standardization of outpatient, short treatment time less medical economic burden, and worth widely used.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Fractures, Closed ; surgery ; Humans ; Male ; Middle Aged ; Radius ; surgery ; Radius Fractures ; surgery ; Splints

DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen (Puhuang) and pine pollen (Songhuafen) samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer (ITS2) from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.
China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Pinus ; classification ; genetics ; Pollen ; classification ; genetics ; Quality Control ; Typhaceae ; classification ; genetics

The study is to establish an HPLC method using fluorescence detector for the determination of doxazosin enantiomers and investigate their chiral inversion in vitro and in vivo. Ultron ES-OVM was taken as the chiral chromatographic column, and the column temperature was 30 degrees C. Isocratic elution using a mobile phase of phosphate buffer-acetonitrile (85 : 15, v/v) at a flow rate of 0.8 mL x min(-1) was done. The fluorescence detection was set at lambda(Ex) = 255 nm and lambda(Em) = 385 nm. Prazosin was used as the internal standard. (-) Doxazosin or (+) doxazosin added into rat plasma in vitro was determined after incubating in 37 degrees C water bath for 2, 5 and 10 days. (-) Doxazosin or (+) doxazosin was administered orally to the rats for one months. Plasma samples were taken at 8 h after the last administration. A good linear relationship was achieved when the concentration of doxazosin enantiomers was within the range of 4 - 2 000 ng x mL(-1). The average recovery for (-) doxazosin was 99.5% with RSD 3.6%, and for (+) doxazosin was 99.3% with RSD 4.3%. Chiral inversion was observed neither in vitro nor in vivo studies. The method is selective, accurate and reproducible, which is suitable for the detection of doxazosin enantiomers in rat plasma. The in vitro and in vivo studies indicate that chiral inversion occurs uneasily between (-) doxazosin and (+) doxazosin in the rat.
Animals ; Blood Chemical Analysis ; methods ; Doxazosin ; blood ; chemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Sensitivity and Specificity ; Stereoisomerism

OBJECTIVETo explore the role of estrogen in the regulation of the expression of claudin-6 and biological behavior in MCF-7 cells.

METHODSRT-PCR and immunocytochemistry were conducted to analyze the expression and localization of claudin-6 in MCF-7 cells treated with 17β-estradiol. CCK-8 kit assay and Scratch Test were conducted to analyze the capability of proliferation and migration of 17β-estradiol treated MCF-7 cells.

RESULTSRT-PCR analysis and immunocytochemistry showed that 17β-estradiol induced a concentration-and time-dependent effect on claudin-6. At 5 nmol/L and at 24 h, 17β-estradiol treatment led to an increased level of claudin-6, which was located in the membrane of MCF-7 cells. CCK-8 analysis showed a significant decrease in the capability of proliferation of MCF-7 cells compared with the control group (P < 0.05). Cells Scratch Test showed decreased migration capability of MCF-7 cells compared with the control group (P > 0.05).

CONCLUSIONS17β-E2 might regulate the expression of claudin-6 and inhibit the proliferation and migration of MCF-7 cells. The inhibitory effects of 17β-E2 on growth and migration of MCF-7 cells may be mediated by claudin-6 expression regulation.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Claudins ; Dose-Response Relationship, Drug ; Estradiol ; administration & dosage ; pharmacology ; Female ; Humans ; Membrane Proteins ; metabolism

The aim of this study was to examine the priming effect of sphingosine 1-phosphate (S1P) on fMLP-activated neutrophils, mainly to detect the neutrophil respiratory burst products, and to investigate the signaling pathway involved in S1P activity. Flow cytometry was used to evaluate the new isolated neutrophil; the superoxide anion output was detected indirectly by cytochrome C reduction in respiratory burst; the dihydro-rhodamine 123 was used to detect the intensity of respiratory burst; the signal transduction pathways of neutrophil respiratory burst were explored by Western blot. The results showed that after pretreated with S1P, the level of superoxide anion released by fMLP-activated neutrophils significantly increased; the Rhodamine 123 mean fluorescence intensity in S1P primed fMLP-activated neutrophils group was significantly higher than that in fMLP treatment group; PI3K and Akt proteins involved in the signal pathway of neutrophil respiratory burst. It is concluded that S1P is a new priming reagent, which primes respiratory burst of fMLP-activated neutrophils; this signal pathway may be that S1P interacts with its receptor, activates PI3K, then activates Akt-transmitting signals through NADPH oxidase, finally results in the respiratory burst.
Cells, Cultured ; Humans ; Lysophospholipids ; metabolism ; NADPH Oxidases ; metabolism ; Neutrophils ; metabolism ; physiology ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptors, Lysosphingolipid ; metabolism ; Respiratory Burst ; Signal Transduction ; Sphingosine ; analogs & derivatives ; metabolism ; Superoxides ; metabolism