OBJECTIVETo observe the curative effects of biliary drainage via endoscope (ENBD) combined with Chinese drug medication on acute biliary tract infection (ABTI) and the influence of treatment on complement 3 (C3), C-reactive protein (CRP) and interleukin-6 (IL-6).

METHODSSixteen patients with ABTI were randomly assigned to two groups: the 9 patients in the combined treatment group (CTG) treated with ENBD combined with Chinese medicine and the 7 patients in routine treatment group (RTG) treated with ENBD alone. Another 18 patients with simple gallbladder stone were taken as the control group (CG). The curative effect was observed and the serum concentrations of C3, CRP and IL-6 were determined before and after treatment.

RESULTSBefore treatment, the concentrations of CRP and IL-6 were significantly higher and C3 lower in all ABTI patients than those in patients with simple gallbladder stone (P < 0.05 or P < 0.01). After treatment, the general condition of patients was improved, the recovery time of intestinal tract function was shortened and the concentrations of C3, CRP and IL-6 significantly decreased in CTG, with the effects better than those in RTG respectively (P < 0.01 or P < 0.05).

CONCLUSIONENBD combined with Chinese drug medication shows favorable curative effects on ABTI. Treatment with Chinese medicine according to syndrome differentiation could decrease blood level of pro-inflammatory cytokines and promote recovery of the injured immune function.

Adult ; Aged ; C-Reactive Protein ; metabolism ; Cholangitis ; blood ; therapy ; Combined Modality Therapy ; Complement C3 ; metabolism ; Drainage ; instrumentation ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Endoscopy, Digestive System ; Female ; Humans ; Interleukin-6 ; blood ; Male ; Middle Aged ; Phytotherapy ; Treatment Outcome

OBJECTIVE: To provide a reference about choosing the methods of isolating exosomes derived from tumor cells including laryngocarcinima Hep-2 cells by comparing advantages and defects of two methods of isolation and extraction exosomes. METHOD: Previously, laryngocarcinoma Hep-2 cells were cultivated massively, then the cells were processed with hot shock in 42 degrees C for 1 h. Sucrose density gradient centrifugation ultrafiltration (method 1): cells culture supernatant 90 ml was gathered, the supernatant was clarified through a 3/0.8 μm small filter to remove impurities and fragments which in larger diameter. Then the filtering fluid was concentrated and purified through sucrose density gradient centrifugation and ultrafiltration, the concentrated fluid was obtained. Exosome Isolation Kit (method 2): cells culture supernatant 4 ml was gathered, the solutions of the kit were added into the supernatant in proper sequence, then filtered by the special column, the concentrated fluid was obtained. Both products are observed by high resolution transmission electron microscopy. RESULT: Both methods could isolate and extract exosomes feasibly. In single high power view of transmission electron microscopy, exosomes of method 1 disperse better, but lower density, and more impurity in background, exosomes of method 2 arrange closer, higher density, and less impurity. CONCLUSION Exosome isolation Kit require less supernatant, cost less time, process procedure briefly, harvest higher yield. It may become a new option of isolating exosomes derived from Laryngocarcinoma Hep-2 cells.
Cell Line, Tumor ; Exosomes ; ultrastructure ; Humans ; Laryngeal Neoplasms ; pathology ; Microscopy, Electron, Transmission

Objective To compare the effects of UVA irradiation on cell morphology,quantity and expression of inducible nitric oxide synthase (iNOS) mRNA and protein in human fibroblasts versus a kerati- nocyte cell line HaCaT.Methods Human fibroblasts and HaCaT cells were cultured and irradiated by 5 J/cm~2 UVA.Then,at 24,48 and 72 h after the stimulation,the cell morphology was observed under an in- verted microscope,and iNOS mRNA and protein were measured by RT-PCR and immunohistochemistry method,respectively.Results After the irradiation,human fibroblasts showed shrinkage at the three time points,the quantities of the cells began to decrease significantly at 24 h (P

The metabolic transformation of the drugs containing carboxylic acid groups can lead to the formation of acyl glucuronide metabolites through catalysis by glucuronosyltransferase, and produce pro-acyl glucuronide intermediate metabolites with electronic activity. Then, protein or DNA adducts appeared after a series of non-enzyme or enzyme reactions. These adducts would change the protein activity and potentially lead to idiosyncratic and genotoxicity. In this paper, we discussed the chemical activity, drug-induced mechanisms, distribution and toxicity resulting from this metabolic activation for these drugs, and stated the status and prospects of research in this field.
Biological Transport, Active ; Biotransformation ; Carboxylic Acids ; metabolism ; toxicity ; DNA Damage ; drug effects ; Drug-Related Side Effects and Adverse Reactions ; Glucuronides ; metabolism ; toxicity ; Glucuronosyltransferase ; metabolism ; Hepatocytes ; metabolism ; Humans ; Pharmaceutical Preparations ; metabolism

OBJECTIVETo evaluate the short-term effect and experience of Nuss procedure on 120 cases of patients with pectus excavatum.

METHODSThoracoscopy assisted Nuss procedure with different ways of anesthesia were applied to 120 cases of patients with pectus excavatum, including 7 cases of recurrence after traditional surgical procedure (6 cases) and Nuss method (another one). The patients ranged in age from 2.5 to 43 (mean 14.1) years and in Haller index from 2.91 to 29. Of the 120, 73 had symmetric and 47 had asymmetric pectus excavatum. The Nuss procedure is performed with general anesthesia and a convex steel bar is inserted under the sternum with thoracoscopy through small bilateral thoracic incisions. The steel bar is inserted with the convexity facing posteriorly, and when it is in position, the bar is turned over, thereby correcting the deformity.

RESULTSThe operation was successfully accomplished without severe complications in all the 120 cases. The mean operative time was 58 minutes and the mean volume of blood loss was 30 ml. 103 patients had one bar inserted while the other 17 cases with more extremely diffuse depression required 2 or even 3 bars to get a satisfactory correction. Such methods as modifications to the fixing points and the shape of the bar, partial osteotomy, were developed to deal with asymmetric ones.

CONCLUSIONThe Nuss procedure is a minimally invasive technique for correction of pectus excavatum. It can lead to a satisfactory outcome and surgical time is less.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Funnel Chest ; surgery ; Humans ; Male ; Orthopedic Procedures ; methods ; Thoracoscopy ; Young Adult

OBJECTIVETo construct a dendritic cell vaccine transduced by murine alpha-fetoprotein (mAFP) gene, and evaluate its immunoprotective effect on C57BL/6J mice during the induction of hepatocellular carcinoma by diethylnitrosamines, carbon tetrachloride and ethanol.

METHODSDendritic cells (DCs) were induced and augmented by murine IL-4 and GM-CSF, and transfected by recombinant adenovirus engineered with mAFP gene. Major MHC class I and II, B7.1 (CD80), B7.2 (CD86), CD18a, and CD54 molecules on DC were analyzed by FACS. 80 C57BL/6J male mice were randomly divided into 4 groups (20 mice per group): Simple DC inoculated group, pAdBM5-mAFP-DC inoculated group, pAdBM5-mAFP plasmid inoculated group, and PBS control group. They were immunized once with 5 x 10(5) DCs (0.1 ml)/mouse administered s. c. in the left flank or 100 mg pAdBMS-mAFP plasmid/mouse administered i. m. in the left tibialis anterior muscle. Inoculation was conducted once a week for 4 weeks after 3 times consecutive immunization initially. At the same time of immunization, DEN/CCl4/ethanol were given to induce hepatocellular carcinoma. Tumor incidence was assessed after 20 weeks.

RESULTSA transgenic DC vaccine was successfully constructed and the mAFP transgenic DCs expressed high level molecules of major MHC class I and II , B7.1, B7.2, CD18a, and CD54. After the 20-week induction, the incidence of primary hepatocellular carcinoma (PLC) was 70.0% in simple DC inoculated group, 25.0% in pAdBMS-mAFP-DC inoculated group, 65.0% in pAdBM5-mAFP plasmid inoculated group, and 75.0% in PBS control group. There was a significant difference between group B and other groups (P < 0.05).

CONCLUSIONmAFP transgenic DC tumor vaccine inoculation may induce strong immunoprotection against liver carcinogenesis and tumor development and reduce PLC incidence induced by DEN/CCl4/ethanol.

Adenoviridae ; genetics ; Animals ; B7-1 Antigen ; metabolism ; Cancer Vaccines ; Carbon Tetrachloride ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; metabolism ; Diethylnitrosamine ; Ethanol ; Genetic Vectors ; Histocompatibility Antigens Class I ; metabolism ; Histocompatibility Antigens Class II ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Liver Neoplasms, Experimental ; chemically induced ; immunology ; prevention & control ; Male ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; alpha-Fetoproteins ; genetics ; metabolism

UNLABELLEDOBJECTIVE To explore the effects of human macrophage inflammatory protein-1 beta (hMIP-1beta) modification on the in vivo tumorigenicity and vaccine efficacy of tumor cells.

METHODSMurine colorectal adenocarcinoma CT26 cells were transfected with a recombinant adenovirus carring the hMIP-1beta gene (AdhMIP-1beta). The efficacy of gene transfection was tested by X-gal staining. The hMIP-1beta level in the supernatant of hMIP-1beta gene-modified CT26 cells was assayed by ELISA, and the chemotactic activity for CD4+ T cells, CD8+ T cells, NK cells and immature dendritic cells (imDCs) were assayed by a transwell chamber. The changes of growth characteristics and in vivo tumorigenicity of hMIP-1beta gene-modified CT26 cells were also assessed. BALB/c mice were immunized with hMIP-1beta gene-modified CT26 tumor vaccine and the antitumor effect was evaluated.

RESULTShMIP-1beta gene could be transfected into CT26 cells by AdhMIP-1beta with an efficiency over 95%. The level of hMIP-1beta in the culture supernatant of hMIP-1beta gene-modified CT26 cells was 980 pg/ml and the supernatant displayed ramarkable chemotactic activity to CD4+ T cells, CD8+ T cells, NK cells and imDCs compared with LacZ gene-modified CT26 cells and control. When the hMIP-1beta gene-modifited CT26 cells were subcutaneously inoculated in BALB/c mice, the tumorigencity was delayed and suppressed, and overt necrosis and lymphocyte infiltration were observed in the tumor tissue, but not in those inoculated with LacZ gene-modified CT26 cells or parental CT26 cells. The mice immunized with hMIP-1beta gene-modified CT26 tumor vaccine could induce tumor specific CTL activity and nonspecific NK activity, and exhibited resistance to later challenge with wild-type CT26 cells.

CONCLUSIONhMIP-1beta gene-modified CT26 cells exhibit decreased tumorigenicity, and hMIP-1beta gene-modified tumor vaccine may induce a powerful specific and nonspecific antitumor response. The data suggested that hMIP-1beta gene-modified tumor vaccine may play a potent role in prevention of metastasis and recurrence of malignant tumors.

Adenocarcinoma ; metabolism ; pathology ; Adenoviridae ; genetics ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Cancer Vaccines ; Cell Line, Tumor ; Chemokine CCL4 ; genetics ; metabolism ; Chemotaxis, Leukocyte ; Colonic Neoplasms ; metabolism ; pathology ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Female ; Genetic Vectors ; Humans ; Killer Cells, Natural ; immunology ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Random Allocation ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden

Hydrophilic low molecular drugs, peptides and proteins, which are always poor in bioavailability, are mainly absorbed through the paracellular way in which the tight junction is the elementary framework. The tight junctions are a multiple unit structure composed of multiprotein complex that affiliates with the underlying apical actomyosin ring. Tight junction proteins are identified including transmembrane proteins (occludin, claudin and JAM) , cytoplasmic plaque proteins (ZO-1, ZO-2, ZO-3 and cingulin) and cytoskeleton. Traditional absorption enhancers can usually impair mucous membranes which constraint the utilization of these enhancers. Recently, with the increasing knowledge of the structure and function of tight junctions, many new absorption enhancers have been developed such as NO donor, CPE, Zot, and so on. In vivo and in vitro studies have shown that these enhancers could be effectively used to increase the absorption of paracellular markers and low bioavailable drug across intestinal epithelium with lower side effect. In short, the transient opening of the tight junctions by these enhancers provides new ideas that could help in novel drug delivery of therapeutic agents.
Animals ; Biological Availability ; Cell Adhesion Molecules ; metabolism ; Cholera Toxin ; pharmacology ; Claudin-1 ; Cytoskeleton ; metabolism ; Decanoic Acids ; pharmacology ; Drug Delivery Systems ; Enterotoxins ; pharmacology ; Humans ; Intestinal Absorption ; drug effects ; Membrane Proteins ; metabolism ; Nitric Oxide Donors ; pharmacology ; Occludin ; Phosphoproteins ; metabolism ; Receptors, Cell Surface ; metabolism ; Tight Junctions ; metabolism ; physiology ; Zonula Occludens-1 Protein

Different factors including hybridization solution components, hybridization temperature, and the concentration and proportion of the labelled primer, which affected the sensitivity and specificity of single mutation identification, were exploited. Asymmetric PCR increased the hybridization sensitivity, and the asymmetric multi-PCR did not affect the specificity, while the sensitivity was improved a little. Among 30 lung cancer samples detected with the oligonucleotide microarray, 12 was found P53 gene mutations and 5 had K-ras gene mutations. The P53 gene mutations identified by the oligonucleotide microarray was proved 80% same as the sequencing results. The obvious statistical relations of K-ras and P53 gene mutations with tumor type, tumor stage and smoking were not obtained because of less samples and mutation sites.
Genes, ras ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; Mutation ; Oligonucleotide Array Sequence Analysis ; methods ; Oligonucleotides ; genetics ; Sensitivity and Specificity ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo investigate the role of 67 000 laminin receptor (LN-R) in the processes of invasion and metastasis of laryngeal squamous cell carcinoma.

METHODSExpression of 67 000 LN-R mRNA in 20 cases with laryngeal squamous cell carcinomas and corresponding normal tissues was determined by RT-PCR, and the relationship between its expression level and patients' clinicopathological features was analyzed. Expression of 67 000 LN-R on the surface of AMC-HN-8 laryngeal carcinoma cells was examined by flow cytometry. The effect of 67 000 LN-R monoclonal antibody (MLuC5) on the adhesive and invasive abilities was observed by adhesion test and Boyden chamber invasiveness test in vitro.

RESULTSThe expression level of 67 000 LN-R mRNA in human laryngeal squamous cell carcinoma was significantly higher than that in normal tissues (P < 0.05). The expression level of 67 000 LN-R mRNA in laryngeal squamous cell carcinomas with cervical lymph node metastases was higher than in those without cervical lymph node metastases (P < 0.05). There was a significant negative correlation between the expression level of 67 000 LN-R mRNA and the degree of tumor differentiation, the level being higher in poorly differentiated tumors (P < 0.05). Flow cytometry showed that (80.9 +/- 0.9)% of AMC-HN-8 cells expressed 67 000 LN-R. MLuC5 inhibited the adhesion of AMC-HN-8 cells on LN, and after treated with MLuC5 for 60 and 120 minutes, the adhesion inhibition rate was 57.1% and 63.2%, respectively. The invasive ability to artificial basement membrane was reduced by MLuC5.

CONCLUSIONLaryngeal carcinoma overexpressing 67 000 LN-R has stronger invasiveness, and 67 000 LN-R monoclonal antibody may contribute to prevent metastasis of laryngeal carcinoma.

Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; immunology ; Carcinoma, Squamous Cell ; metabolism ; secondary ; Cell Adhesion ; immunology ; Cell Differentiation ; Cell Line, Tumor ; Female ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Laminin ; biosynthesis ; genetics ; immunology