Astrocytoma ; genetics ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Frizzled Receptors ; genetics ; metabolism ; Glioblastoma ; genetics ; metabolism ; Glioma ; genetics ; metabolism ; Humans ; Paraffin Embedding ; RNA, Messenger ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Signal Transduction ; Wnt2 Protein ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

Objective To assese the healing of stoma after magnetic anastomosis for the reconstruction of biliary-enteric continuity under severe inflammation.
Methods Acute bile duct injury was constructed as a bile peritonitis model in mongrel dogs (n=32). Magnetic anastomosis (group A, n=16) and traditional suture anastomosis (group B, n=16) were performed to reconstruct the biliary-enteric continuity in one stage. Half of the dogs in each group were euthanized on the 30th postoperative day, and the other half on the 90th postoperative day to harvest the stoma region. The healing conditions of the stoma after the 2 anastomotic approaches were observed with naked eyes, under light microscope and scanning electron microscope.
Results The stoma leakage rate (50%versus 0%on the 30th postoperative day, 37.5%versus 12.5%on the 90th postoperative day, both P<0.05) and stenosis degree (13.9%±0.3%versus 7.1%±0.3%on the 30th postoperative day, 17.2%±0.4%versus 9.4%±0.4%on the 90th postoperative day, both P<0.01) were significantly higher in group B than in group A. Compared with traditional manual anastomoses, the histological analysis under light and electron microscope showed a more continuous stoma with more regular epithelium proliferation and collagen arrangement, less inflammation in group A.
Conclusions Magnetic anastomosis stent ensures better healing of the stoma even under the circumstance of severe inflammation.

Adult ; Female ; Humans ; Liver ; immunology ; pathology ; Liver Cirrhosis, Biliary ; immunology ; pathology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology

To compare the DAAO expression level in different Pichia pastoris host strains, the gene encoding DAAO from Trigonopsis variabilis was cloned into plasmid pPIC3.5k and then transformed into P. pastoris GS115 and KM71 respectively. The positive transformants PDK13 (MutS) and PD27 (Mut+) were obtained by PCR analysis. Their optimal and different expression conditions were investigated. To compare with PD27, PDK13 was determined to poss a slower consumption of methanol, a longer induction time, a lower oxygen request and apparently higher expression of DAAO. The highest expression levels were reached up to 2700, 2500 IU/L in shaking flask and 10140, 8463.5 IU/L in fermentor respectively. The over-expression of DAAO can meet its large demand for production of 7-ACA, alpha-keto acid and L-amino acid. In addition, the phenylpyruvate and L-phenylalanine were obtained by crude DAAO reacting with DL-phenylalanine at 37 degrees C for 3h.
D-Amino-Acid Oxidase ; genetics ; Fermentation ; Methanol ; metabolism ; Phenylalanine ; metabolism ; Pichia ; genetics ; Polymerase Chain Reaction

Esophageal Neoplasms ; diagnosis ; pathology ; Humans ; Liposarcoma ; diagnosis ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Tomography, X-Ray Computed

OBJECTIVETo investigate the possible roles of adenosine and the cytokines TNF-alpha and IL-10 in the pathogenesis of acute bacterial prostatitis (ABP) in rats.

METHODSForty-eight male Wistar rats were randomly divided into groups A (ABP), B (ABP + theophylline intervention), C (sham) and D (blank control). ABP models were established by injecting Escherichia coli 0157 into the prostate, and those in group B were treated by intraperitoneal injection of theophylline immediately after modeling. At 4 and 14 days, the prostate tissues of the rats were collected for detection of the expressions of TNF-alpha and IL-10 by immunohistochemistry and the concentration of adenosine by high-performance liquid chromatography.

RESULTSAt 4 and 14 days, the concentrations of adenosine were significantly higher in group A ([48.38 +/- 17.27] and [26.54 +/- 11.22] microg/g) than in C ([0.45 +/- 0.25] and [0.46 +/- 0.29] microg/g) and D ([0.41 +/- 0.23] and [0.43 +/- 0.27] microg/g) (P < 0.05), and so were the expressions of TNF-alpha in A (0.23 +/- 0.08 and 0.21 +/- 0.03) than in C (0.07 +/- 0.03 and 0.07 +/- 0.01) and D (0.07 +/- 0.06 and 0.07 +/- 0.06) (P < 0.05), and those of IL-10 in A (0.13 +/- 0.03 and 0.25 +/- 0.01) than in C (0.07 +/- 0.03 and 0.07 +/- 0.03) and D (0.07 +/- 0.01 and 0.07 +/- 0.02) (P < 0.05). Compared with group A, the rats in group B showed significant increases at 4 and 14 days in the severity of inflammation, concentration of adenosine ([86.64 +/- 32.87] and [51.17 +/- 22.96] microg/g, P < 0.05) and expression of TNF-alpha (0.37 +/- 0.08 and 0.32 +/- 0.06, P < 0.05), but exhibited no remarkable difference in the expression of IL-10 (0.12 +/- 0.06 and 0.15 +/- 0.06, P > 0.05).

CONCLUSIONAdenosine may affect the progression of inflammation by regulating the expressions of the cytokines TNF-alpha and IL-10 in ABP rats through the adenosine receptor signaling pathway.

Adenosine ; physiology ; Animals ; Escherichia coli O157 ; Interleukin-10 ; metabolism ; Male ; Prostate ; drug effects ; metabolism ; Prostatitis ; metabolism ; microbiology ; Random Allocation ; Rats ; Rats, Wistar ; Theophylline ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

Human transferrin receptor (TfR) was isolated from homogenates of placental tissues by affinity chromatography on transferrin-Sepharose, and then used to screen human scFv against it from a fully-synthesized phage scFv library. After verifying the specificity, gene fragment of one of the selected scFv was inserted into the plasmid pET22b(+) and transformed into E. coli BL21(DE3) . Expression of scFv in transformant was induced with 0.5mmol/L IPTG. ELISA assay on HeLa cells showed that scFv protein could recognize and bind to TfR on the surface of HeLa cells. The scFv was purified by one-step affinity chromatography with Ni+ -NTA agarose, and injected into Kunming mouse via tail veins. This scFv was detected in brain tissues 1h later by capillary depletion method, which indicates that scFv protein can permeate through the blood brain barrier by mediation of the TfR receptor. Our works lay the foundation for the treatment of tumors and central nervous system diseases.
Animals ; Antibodies, Anti-Idiotypic ; genetics ; isolation & purification ; metabolism ; Escherichia coli ; genetics ; metabolism ; HeLa Cells ; Humans ; Immunoglobulin Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; immunology ; Mice ; Receptors, Transferrin ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; Transferrin ; metabolism

BACKGROUNDCytosine deaminase (CD) converts 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU) in CD/5-FC gene therapy, 5-FU will be mostly converted into nontoxic beta-alanine without uracil phosphoribosyltransferase (UPRT). UPRT catalyzes the conversion of 5-FU to 5-fluorouridine monophosphate, which directly kills CD::UPRT-expressing cells and surrounding cells via the bystander effect. But the pharmacokinetics and the bystander effect of CD::UPRT/5-FC has not been verified in vivo and in vitro. Before the CD::UPRT/5-FC bi-gene therapy system is used in clinical trial, it is essential to monitor the transgene expression and function in vivo. Thus, we developed a preclinical tumor model to investigate the feasibility of using (19)F-magnetic resonance spectroscopy ((19)F-MRS) and optical imaging to measure non-invasive CD and UPRT expression and its bystander effect.

METHODSC6 and C6-CD::UPRT cells were cultured with 5-FC. The medium, cells and their mixture were analyzed by (19)F-MRS. Rats with intracranial xenografted encephalic C6-CD::UPRT glioma were injected intraperitoneally with 5-FC and their (19)F-MRS spectra recorded. Then the pharmacokinetics of 5-FC was proved. Mixtures of C6 and C6-CD::UPRT cells at different ratios were cultured with 5-FC and the cytotoxic efficacy and survival rate of cells recorded. To determine the mechanism of the bystander effect, the culture media from cell comprising 25% and 75% C6-CD::UPRT cells were examined by (19)F-MRS. A comparative study of mean was performed using analysis of variance (ANOVA).

RESULTS(19)F-MRS on samples from C6-CD::UPRT cells cultured with 5-FC showed three broad resonance signals corresponding to 5-FC, 5-FU and fluorinated nucleotides (F-Nuctd). For the C6 mixture, only the 5-FC peak was detected. In vivo serial (19)F-MRS spectra showed a strong 5-FC peak and a weak 5-FU peak at 20 minutes after 5-FC injection. The 5-FU concentration reached a maximum at about 50 minutes. The F-Nuctd signal appeared after about 1 hour, reached a maximum at around 160 minutes, and was detectable for several hours. At a 10% ratio of C6-CD::UPRT cells, the survival rate was (79.55 +/- 0.88)% (P < 0.01). As the C6-CD::UPRT ratio increased, the survival rate of the cells decreased. (19)F-MRS showed that the signals for 5-FU and F-Nuctd in the culture medium increased as the ratio of C6-CD::UPRT in the mixture increased.

CONCLUSIONS(19)F-MRS studies indicated that C6-CD::UPRT cells could effectively express CD and UPRT enzymes. The CD::UPRT/5-FC system showed an obvious bystander effect. This study demonstrated that CD::UPRT/5-FC gene therapy is suitable for 5-FC to F-Nuctd metabolism; and (19)F-MRS can monitor transferred CD::UPRT gene expression and catalysis of substrates noninvasively, dynamically and quantitatively.

Animals ; Antimetabolites ; pharmacokinetics ; therapeutic use ; Cell Line ; Cytosine Deaminase ; genetics ; physiology ; Flucytosine ; pharmacokinetics ; therapeutic use ; Genetic Therapy ; methods ; Glioma ; drug therapy ; therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Pentosyltransferases ; genetics ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the associated risk factors of metabolic syndrome in Jiangsu province, China.

METHODSUsing identical protocol and questionnaire, an epidemiological study was carried out in a population of 5888 adults in 12 counties in Jiangsu. Anthropometric test and blood sampling were conducted at the time of interview. IDF (2005) was used as the diagnostic criteria of metabolic syndrome . The prevalence and age-standardized prevalence of metabolic syndrome were calculated. Univariate and multivariate logistic regression model were used to identify associated risk factors.

RESULTSThe prevalence and age-standardized prevalence of metabolic syndrome in Jiangsu were 19.07% (11.10% in males and 25.72% in females) and 17.48% (11.49% in males, 22.86 % in females), respectively. Among the potential risk factors of metabolic syndrome as gender, age, education level, occupation, income, physical activity, smoking, alcohol drinking, disease family history, data from univariate and multivariate logistic regression analyses suggested that gender (OR = 1.91), age (OR = 1.15), physical inactivity (OR = 1.94), with hypertension family history (OR = 1.99) and with obesity family history (OR = 6.24) could significantly increase the risk of disease development.

CONCLUSIONMetabolic syndrome has become a significant public health problem among the adults in Jiangsu province.

Adult ; Age Factors ; Aged ; Alcohol Drinking ; adverse effects ; China ; epidemiology ; Female ; Humans ; Logistic Models ; Male ; Metabolic Syndrome ; blood ; epidemiology ; genetics ; Middle Aged ; Motor Activity ; physiology ; Risk Factors ; Sex Factors ; Smoking ; adverse effects ; Surveys and Questionnaires

OBJECTIVETo investigate the effects of topical application of small dose of insulin on the wound healing of the scalded rats, so as to explore its mechanism.

METHODSThe rats employed in the study were subjected to deep partial thickness burn and were divided into group A (with subcutaneous injection of isotonic saline into the rat wounds as control), B and C (with subcutaneous injection of 0.1 U and 1 U insulin in the rat wounds respectively) and D (with subcutaneous injection of 0.1 U insulin in the rat abdomen as control). The wound healing time and wound healing rate were assessed every other day after 3 postburn days (PBDs). The histological changes of the wounds after injection were examined, the changes in the cell cycle of epidermal cells in the wound were analyzed by flow-cytometry, and blood glucose concentration of each group was determined.

RESULTSThe wound healing time in group B (18.36 +/- 4.12 d) was significantly shorter than that in other groups (A: 24.57 +/- 5.19 d, C: 21.46 +/- 2.97 d, D: 24.50 +/- 1.05 d, P < 0.01). The wound healing rate of the rats in group B in 5, 9, 11, 13, 15, 17 and 19 PBD was obviously higher than that in group A, and was markedly higher than that in group C on 17 PBD (P < 0.05 - 0.01). The epithelial layer was thinner with less epidermal nails but much more fibroblasts in epidermal layer in group A, while the epithelial layer was thicker with abundant epidermal nails in group B and C with many fibroblasts in the dermis. The amount of cells in S phase at 4 PBD in group B was dramatically higher than that in group A, and cells in G2M phase at 4 - 5 PBD in group B was also higher than that in group A and C (P < 0.05 - 0.01). The blood level of glucose in group A and B fluctuated between 3.42 to 4.62 mmol/L at 24 PBH, while that in group C and D decreased obviously 1 hour after injection (P < 0.01), but gradually returned to normal 4 hours after injection.

CONCLUSIONLocal injection of small dose of insulin may accelerate burn wound healing due to its role in promoting the proliferation and division of the repairing cells.

Administration, Topical ; Animals ; Blood Glucose ; analysis ; Burns ; blood ; drug therapy ; physiopathology ; Cell Cycle ; drug effects ; Female ; Insulin ; administration & dosage ; Male ; Rats ; Rats, Sprague-Dawley ; Wound Healing ; drug effects