BACKGROUNDNeuroblastoma, one of the common tumors in children, possesses the feature of natural regression that might be related to apoptosis caspase-3 and survivin are believed to respectively induce and inhibit apoptosis. We investigated the expression of caspase-3 and survivin in pediatric neuroblastoma and the role that these genes played in apoptosis.

METHODSThe expression of caspase-3 and survivin in pediatric neuroblastoma tissue samples was detected using in situ hybridization, ter mintuesal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and immunohistochemical staining. The role that these genes played in apoptosis was then evaluated.

RESULTSA converse correlation was observed between the expression of survivin and caspase-3. When survivin was expressed at high levels in neuroblastoma samples, caspase-3 expression was downregulated, and the apoptotic index decreased simultaneously.

CONCLUSIONThere is a converse correlation between the expression of caspase-3 and the expression of survivin in neuroblastoma cells, indicating that caspase-3 might induce apoptosis, and survivin may inhibit this process.

Apoptosis ; Caspase 3 ; Caspases ; analysis ; physiology ; Female ; Humans ; Immunohistochemistry ; Infant ; Infant, Newborn ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; analysis ; physiology ; Neoplasm Proteins ; Neuroblastoma ; chemistry ; pathology

OBJECTIVETo study the enhancing effect of isoflavonoid genistein in irradiation (IR) on prostate DU145 cancer cells.

METHODSProstate cancer cell line DU145 was used in this experiment. Clonogenic assay was applied to compare the survival fractions of DU145 cells after treatments with genistein alone and/or graded IR. DNA electrophoresis and TUNEL method were applied to detect cell apoptosis. Cell cycle was observed using flow cytometry and related protein expressions by immunoblotting.

RESULTClonogenic assay demonstrated that genistein, even at low to medium concentrations, enhanced the radiosensitivity of DU145 cells. After treatments with IR and/or genistein for 24 h, apoptosis was mainly seen with genistein at high concentration and was minimally dependent on IR. Apoptosis also occurred after treatments for 72 h with lower concentrations of genistein, especially when combined with IR. While IR or genistein led to a G2/M cell cycle arrest, combination of them could further increase DU145 cells at G2/M phase. This G2/M arrest was largely maintained at 72 h, and accompanied by increasing apoptosis and hyperdiploid cell populations. Cell-cycle related protein analysis disclosed biphasic changes in cyclin B1, less markedly increased cdc-2 and stably elevated p21(cip1) levels with increasing genistein concentrations.

CONCLUSIONGenistein could enhance the radiosensitivity of DU145 prostate cancer cells. The mechanisms might be involved in the increased apoptosis, prolonged cell cycle arrest and impaired damage repair induced by the combined treatment.

Apoptosis ; drug effects ; radiation effects ; CDC2 Protein Kinase ; analysis ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Cyclin B ; analysis ; Cyclin B1 ; G2 Phase ; drug effects ; radiation effects ; Genistein ; pharmacology ; Humans ; Male ; Prostatic Neoplasms ; pathology ; radiotherapy ; Radiation-Sensitizing Agents ; pharmacology ; S Phase ; drug effects ; radiation effects

In this study we present our experiences with the reverse sural fasciocutaneous flap to reconstruct the distal lower limb soft tissue defects caused by traumatic injuries. These flap graftings were carried out from Oct. 2010 to Dec. 2012 in our department. The series consisted of 36 patients, including 21 men and 15 women with an average age of 46.2 years (14-83 years) and with a medium follow-up period of 18 months (12-24 months). Of all the cases of acute trauma, there were 10 cases of trauma of distal tibia, 9 cases of trauma of perimalleolus, and 17 cases of trauma of midfoot and forefoot. Related risk factors in the patients were diabetes (2 cases), advanced age (>65 years, 3 cases) and cigarette smoking (6 cases). The reverse flow sural island flap irrigation depended on lower perforators of the peroneal artery. The fasciocutaneous pedicle was 3-4 cm in width and the anatomical structures consisted of the superficial and deep fascia, the sural nerve, short saphenous vein, superficial sural artery together with an islet of subcutaneous cellular tissue and skin. The most proximal border of the flap was only 1.5 cm away from the popliteal skin crease and the pivot point was 5-7 cm above the tip of the lateral malleolus. All the flaps survived. No arterial crisis occurred in any case. The venous congestion occurred in 2 cases and got better after raising the limbs and bloodletting. Only in an old man, 1.5 cm necrosis of distal margin of his flap occurred and finally healed after continuous dressing change. One-stage skin grafting was performed, and all the donor sites were sutured and successfully healed. It was concluded that the reverse sural fasciocutaneous flap is safe and reliable to extend to the proximal third even near the popliteal skin crease. We also concluded this flap can be safely and efficiently used to treat patients with large and far soft-tissue defects from the distal leg to the forefoot with more versatility and it is easier to reach the recipient sites.

The gene coding for beta-glycosidase (EC3.2.1.21) from Thermus nonproteolyticus HG102 has been cloned and expressed in E. coli. The gene open reading frame was 1311 bp and it codes for 436 amino acids. The deduced amino acid sequence of the enzyme showed identity with members of glycosyl hydrolase family I. The enzyme had high content of hydrophobic amino acid (Ala 12.8%, Leu 10.9%), Arg(9.6%), Glu(9.4%) and Pro(8.0%), but low content Cys(0.45%) and Met (0.9%). From the alignment of enzyme amino acid sequence with other glycosyl hydrolase family I members, Glu164 and Glu338 were predicated as the proton donor and nucleophile group. The DNASTAR program was used to predict the secondary structure. According to the Chou-Fasman model, the enzyme has 41.4% of alpha-helics, 16.2%, beta-strands, 14.4%, beta-turns. 14 of the 35 Pro were located at the second sites of beta-turns. Hydrophobic interaction, ion bond, alpha-helics and Pro had important contribution to Tn-gly thermostability.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; Glycoside Hydrolases ; biosynthesis ; classification ; genetics ; Hot Temperature ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Phylogeny ; Protein Structure, Secondary ; physiology ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA ; methods ; Sequence Homology ; Thermus ; enzymology ; beta-Glucosidase

OBJECTIVETo investigate the initial colonization of Mutans streptococci (MS) on teeth of children.

METHODSDental plaque was collected from 123 children aged from 7 to 42 months. MS was isolated by culturing on selective medium and identified by biochemical tests.

RESULTSMS was detected in 51 of 123 children. The rate of colonization was 41.5%, and the mean age of colonization was 29 month. The incidence of MS infection increased with age and tooth eruption, and reached peak at 25 - 31 months, which was statistically significant. There was a correlation between infection of MS and dental caries.

CONCLUSIONThe results indicate that the susceptive period of MS infection is from 25 to 31 months when the second primary molars erupt. MS infection is associated with dental caries.

Age Factors ; Child, Preschool ; Dental Caries ; microbiology ; Dental Plaque ; microbiology ; Female ; Humans ; Infant ; Male ; Streptococcus mutans ; growth & development ; Tooth Eruption

Objective　To study the role of TNFα in the plasm a and skin and IL-1 in the serum in the formation of secondary thrombosis after skin avulsion. Methods　After avulsive flap at size of 12 cm×4 cm was inflicted on the hind legs of pigs, skin specimens and venous blood sam ples were taken at various time points. The contents of TNFα in plasma and skin were determined with radio-immunoassay, and the activity of serum IL-1 wi th 3［H］-TdR. Results　The TNFα contents in the plasma and skin were increased significantly after avulsion（P<0.01），which were (41 5±24) ng/L and (298±18.5) ng/L respectively on the 3rd day after the injury. T he activity of IL-1 in the serum was increased (P<0.05) and was (2.59± 0.85 ) ng/L on day 3. Conclusion　The changes of TNFα contents and I L-1 activity in blood and skin play important roles in the inducetion and aggra vation of secondary tissue necrosis and early thrombosis after skin avulsion.

Objective　To investigate the molecular regulation of G1 arrest of mouse thymocytes induced by ionizing radiation. Methods　Cell cycle was analyzed by flow cytometry (FCM) following staining of cells with proidium iodide. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression. Results　It was demonstrated that G1 phase of mouse thymocytes increased significantly at 12h after whole body irradiation (WBI) with the doses of 0.5, 1.0 and 2.0 Gy, and at 24h following 2.0Gy exposure, measured by FCM. In the time course experiment, it was found that G1 phase of thymocytes increased significantly at 4h, reached a peak level at 24h and came down toward 48h after WBI with 2.0Gy X-rays. The results also showed that after 2.0Gy exposure, the expression of proteins in mouse thymocytes increased significantlty from 1h to 8h for p53, for p21 from 4h to 48h, and for MDM2 at 4h and 8h, measured by FCM. But no change was found for GADD45 protein expression. Conclusion　These results suggest that G1 arrest could be induced by a single dose of 0.5 Gy, 1.0Gy or 2.0Gy, and its molecular control might be established through the p53-p21 pathway.

Biomarkers ; analysis ; Child ; Humans ; Mass Spectrometry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Wilms Tumor ; metabolism

AIMTo assess the antioxidative properties and the mechanism of action of dihydromyricetin (DMY) from Ampelopsis grossedentata.

METHODSThe antioxidative properties of DMY were measured by scavenging 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and inhibiting lipid peroxidation induced by FeSO4-edetic acid in linoleic acid. The mechanism of antioxidative properties of DMY was tested by measuring the chelating activities of DMY for Fe2+ with ultraviolet spectrum (UV) method.

RESULTSThe specific absorption of DPPH radical solution at 517 nm was reduced 73.3%-91.5% when DMY was added into the reaction solution in the concentration range from 0.01% to 0.04%. DMY was shown to greatly inhibit the increase of lipid peroxidation (LPO) values in linolei acid system catalyzed by FeSO4-edetic acid. The reaction rates (A532.min-1) of lipid peroxidation were 0.0021-0.0004 in the concentration range from 0.01% to 0.04% and the inhibition activities of DMY was found to be in a concentration-dependent manner. The mechanism of antioxidative properties of DMY was chelating Fe2+ in the Fe(2+)-dependent lipid peroxidation system.

CONCLUSIONDMY showed great antioxidative effect and would be a good natural antioxidant.

Ampelopsis ; chemistry ; Antioxidants ; isolation & purification ; pharmacology ; Chelating Agents ; pharmacology ; Flavonols ; chemistry ; isolation & purification ; pharmacology ; Free Radical Scavengers ; isolation & purification ; pharmacology ; Lipid Peroxidation ; drug effects ; Molecular Structure ; Plants, Medicinal ; chemistry

AIMTo study the effect of the combined use of genistein and ionizing radiation (IR) on prostate DU145 cancer cells.

METHODSDU145, an androgen-independent human prostate cancer cell line, was used in the experiment. Clonogenic assay was used to compare the survival of DU145 cells after treatments with genistein alone and in combination with graded IR. Apoptosis was assayed by DNA ladder and TUNEL stain. Cell cycle alterations were observed by flow cytometry and related protein expressions by immunoblotting.

RESULTSClonogenic assay demonstrated that genistein, even at low to medium concentrations, enhanced the radiosensitivity of DU145 cells. Twenty-four hours after treatment with IR and/or genistein, apoptosis was mainly seen with genistein at high concentrations and was minimally related to IR. At 72 h, apoptosis also occurred in treatment with lower concentration of genistein, especially when combined with IR. While both IR and genistein led to G2/M cell cycle arrest, combination of them further increased the DU145 cells at G2/M phase. This G2/M arrest was largely maintained at 72 h, accompanied by increasing apoptosis and hyperdiploid cell population. Cell-cycle related protein analysis disclosed biphasic changes in cyclin B1 and less dramatically cdc-2, but stably elevated p21 cip1 levels with increasing genistein concentrations.

CONCLUSIONGenistein enhanced the radiosensitivity of DU145 prostate cancer cells. The mechanisms might be involved in the increased apoptosis, prolonged cell cycle arrest and impaired damage repair.

Androgens ; physiology ; Anticarcinogenic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; DNA, Neoplasm ; analysis ; biosynthesis ; genetics ; Flow Cytometry ; Genistein ; pharmacology ; Humans ; Immunoblotting ; In Situ Nick-End Labeling ; Male ; Prostatic Neoplasms ; drug therapy ; radiotherapy ; Tumor Stem Cell Assay