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 study Chinese medicine treatment in the three-part of the proximal humerus fractures.

METHODSFrom January 2009 to February 2012, 118 cases of proximal humerus three-part fractures were used two methods of operation and manipulation treatment,that were all acute and closed. In operation group: there were 22 males and 37 females,the mean age of the patients was (65.80 +/- 10.62) years (ranged from 45 to 83 years), and the interval from injury to hospital was (22.58 +/- 22.11) hours (ranged from 1 to 96 hours), used open reduction and locking plate fixation surgery. In manipulation group: there were 21 males and 38 females, the mean age of the patients was (65.98 +/- 11.10)years (ranged from 45 to 85 years), and the interval from injury to hospital was (20.85 +/- 22.63) hours (ranged from 1 to 107 hours), used manipulative reduction and small splinting external fixation. All patients were evaluated with shoulder pain, function, activity and anatomical indicators after treatment.

RESULTSAll patients were followed up for 3 to 12 months with an average of 8.2 months. According to Neer Score, the total scores was 85.47 +/- 6.15 in operation group, 84.95 +/- 5.70 in manipulation group. The satisfaction rate of the operation group were 88.20%, and the manipulation group were 86.40%. The difference was not statistically significant between two groups (P > 0.05).

CONCLUSIONThe two treatment were able to achieve satisfactory results. The manipulative reduction and splinting treatment has the advantage of avoiding the risk of surgery, less blood damage, ensureing the efficacy, and reducing costs. It can effectively treat the proximal humerus three-part fracture.

Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Follow-Up Studies ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Shoulder Fractures ; therapy ; Splints

OBJECTIVETo determine an optimal insemination technique for patients suspected of high risk of fertilization failure and undergoing assisted reproduction treatment.

METHODSNinety-nine couples were treated by conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in one cycle (half-ICSI) by dividing the sibling oocytes in halves. The clinical and laboratory data were analyzed, and the rates of fertilization, cleavage, good embryos and clinical pregnancy were compared between different fertilization methods.

RESULTSIn the half-ICSI group, the fertilization rate of ICSI (80.5%) was significantly higher than that of IVF (42.9%) (P < 0.01), and so were the rates of complete fertilization failure (21.2%) and low fertilization (16.2%) of IVF than those of ICSI (0 and 3.0%). No significant differences were observed in the rates of cleavage and good-quality embryos between the two groups (P > 0.05).

CONCLUSIONICSI can help to avoid complete fertilization failure, achieve more high quality embryos for transfer and improve the rate of pregnancy for patients with high risk of fertilization failure.

Female ; Fertilization in Vitro ; methods ; Humans ; Male ; Oocytes ; cytology ; Pregnancy ; Pregnancy Outcome ; Sperm Injections, Intracytoplasmic ; methods

AIMTo study compounds isolated from Picria fel-terrae.

METHODSThe chemical constituents were separated and purified by column chromatography on silica gel and MCI. Their structures were identified on the basis of spectral data (IR, UV, MS, ID NMR and 2D NMR).

RESULTSA new cucurbitacin, along with a known one, were obtained from the 60% EtOH extract of the whole plant.

CONCLUSIONThe new compound was identified as 11, 24-dioxo-5, 21-diene-cucurbit-3alpha-O-beta-D-xylopyranosyl-16alpha-O-alpha-L-rhamnopyranoside (dehydrobryogenin glycoside). The known one, hexanorcucurbitacin F, was obtained for the first time from Picria fel-terrae.

Molecular Structure ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification ; Scrophulariaceae ; chemistry ; Steroids ; chemistry ; isolation & purification

OBJECTIVETo compare the effect of the extracts from Decoction for resuscitation (DRE) and its component herbs on prostacyclin (PGI2), thromboxane A2 (TXA2) and nitric oxide (NO) release from rat vascular endothelial cells under hypoxia.

METHODAfter treatment with the extracts from DRE and its component herbs, the contents of 6-keto-prostaglandin F1alpha(6-keto-PGF1alpha), thromboxane B2 (TXB2) as well as nitrite (NO), which were degradation products of PGI2, TXA2 and NO respectively, in culture medium of rat vascular endothelial cells under hypoxia were measured with radioimmunoassay and Griess Reaction.

RESULTCompared with the control group, the results indicated that DRE, prepared licorice root extract (LE), dried ginger extract (GE), aconite root extract (AE), extracts of aconite root and prepared licorice root (ALE), extracts of aconite root and dried ginger (AGE) increased significantly the content of 6-keto-PGF1alpha and the ratio of 6-keto-PGF1alpha/TXB2, but had no effect on the content of TXB2 in culture medium of rat vascular endothelial cells under hypoxia. The content of 6-keto-PGF1alpha in the DRE group was higher than that in the groups of LE, GE, AE, ALE, AGE. The ratio of 6-keto-PGF1alpha/TXB2 in the DRE group was higher than that of the groups of GE, AE, ALE. Compared with the control group, DRE, LE, GE, AE, ALE, AGE increased significantly the content of NO2- in culture medium of rat vascular endothelial cells under hypoxia. Moreover, the content of NO2- in the DRE group was higher than that of the groups of GE, AE, ALE.

CONCLUSIONThe results suggested that DRE increased significantly the content of PGI2 and the ratio of PGI2/TXA2 as well as the content of NO. The effect of DRE on the parameters in culture medium of rat vascular endothelial cells under hypoxia was better than that of the extracts from its component herbs.

6-Ketoprostaglandin F1 alpha ; metabolism ; Aconitum ; chemistry ; Animals ; Aorta, Abdominal ; cytology ; Cell Hypoxia ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; metabolism ; Ginger ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Thromboxane B2 ; metabolism

OBJECTIVETo evaluate the effect of cluster of differentiation 14 (CD-14) and Toll like receptors (TLR) on the expression of interleukin-6 (IL-6) mRNA induced by Porphyromonas endodontalis (Pe) lipopolysaccharides (LPS).

METHODSMC3T3-E1 cells were treated with 10 mg/L Pe-LPS for different hours, and the cells uninvolved by anything as the blank group. The expression of IL-6 was detected by reverse transcription polymerse chain reaction (RT-PCR) and enzyme-liked immunosorbent assay (ELISA). The expression of CD-14, TLR-2 and TLR-4 mRNA was observed at different time point (0 - 24 h) by RT-PCR. The protein of CD-14, TLR-2 and TLR-4 was analyzed with a flow cytometer. MC3T3-E1 cells were pretreated with anti-CD-14, anti-TLR-2 and anti-TLR-4 antibody for 1 h, and then cells were stimulated with 10 mg/L Pe-LPS for 6 h. The expression of IL-6 mRNA was examined by RT-PCR. Statistical analysis was performed using one-way ANOVA Dunnett-t test with SPSS 11.0 software package.

RESULTSThe IL-6 mRNA and proteins increased significantly after treatment with Pe-LPS. When MC3T3-E1 cells treated by Pe-LPS for 6 h, the expression of proteins soared from (11.696 ± 0.672) ng/L to (36.534 ± 0.574) ng/L (P < 0.01); In the control group, the CD-14 and TLR-4 mRNA are ambly-expression, and the ratios of CD-14 and TLR-4 positive cells were (39.038 ± 3.131)% and (11.438 ± 0.385)% respectively in MC3T3-E1. After treatment by Pe-LPS, the expression of CD-14 and TLR-4 mRNA increased significantly, and the ratios of CD-14 and TLR-4 positive cells markedly increased to (62.407 ± 1.800)% and (21.367 ± 2.271)%. TLR-2 expression did not change apparently after Pe-LPS treatment. The expression of IL-6 mRNA was partly inhibited by anti-CD-14 or anti-TLR-4 antibody, but not by TLR-2.

CONCLUSIONSPe-LPS can induce the expression of IL-6 in osteoblast MC3T3-E1 through CD-14 and TLR-4, but not TLR-2.

3T3 Cells ; Animals ; Antibodies ; immunology ; Interleukin-6 ; genetics ; metabolism ; Lipopolysaccharide Receptors ; genetics ; metabolism ; Lipopolysaccharides ; isolation & purification ; pharmacology ; Mice ; Porphyromonas endodontalis ; RNA, Messenger ; metabolism ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism