Objective To detect the regional genomic instability of B16 cells treated with 60Co γ-rays by a green fluorescence protein (GFP)-based genomic instability reporting system.Methods Three groups were employed as non-transfection group,vector control group and transfection group.The GFP-marked reporter construct pCMV-EGFP2XhoI for regional genomic instability was successfully transfected into B16 cells using liposome.B16 cells were selected by screening of G418 with a series of concentrations and limiting dilution cultures to yield a single colony.B16 cells with the genomic instability report system were then irradiated by 60Co γ-rays at doses of 0,2 and 4 Gy.The regional genomic instability of B16 cellswas quantified by counting the number of cells with GFP expression.Results B-16 cell strain steadilyexpressing the GFP-based genomic instability reporting system was established successfully.GFP-positiveB16 cells were observed at 1 d after irradiation with 60Co γ-rays at doses of 2 and 4 Gy.Positive correlations between fluorescence intensity and dose and fluorescence intensity and time were also observed.The positive expression rate of GFP followed the increased of dose (F =36.55,36.76,P ＜ 0.05) and time (t =-3.27,-3.16,-4.26,-6.11,-7.17,P ＜ 0.05),and differences between groups were significant.The positive expression rate of GFP increased significantly at 3 d,and maximum expression was observed at 5 d(2.46 ± 0.24 and 3.82 ± 0.35).The level was tending towards stability.Spontaneous GFP expression at a ratio of 1/600 000 was observed in 0 Gy group after 2 weeks of culture.Conclusions The regional genomic instability of B16 cells induced by 60Co γ-rays can be detected using a GFP-labelled genomic instability reporter system.

0.05). Conclusions The therapeatic effect of breast-conserving therapy of stage I,and IIa breast cancer located ≥3cm of the nipple is similar to that of modified radical mastectomy. Breast-conserving therapy can gradually become the operation of first choice for stage I and IIa breast cancer.

OBJECTIVETo observe the effect of Gansui Banxia Tang plus-minus Gansui and Gancao anti-drug combination on hepatic and renal functions in malignant ascites rats to explore whether the efficacy or toxicity associated with the anti-drug combination.

METHODThe male wistar rats were randomly divided into a blank group, model group, furosemide group, Gansui Banxia Tang group, Gansui Banxia Tang removed Zhigancao group, Gansui Banxia Tang removed Cugansui group, Gansui Banxia Tang removed Zhigancao and Cugansui group. In addition to normal feeding, every morning except for the blank group and model group, the rest of the group was given drugs, the control group and the model group was given distilled water, the volume is 10 mL x kg(-1). Administered five days, all rats were fasted but except water for 24 hours to collect urine. Administered nine days all rats were fasted but except water for 12 hours, we need to weigh weight of rats. When we remove the ascites, we also need to weigh weight of rats. We use the weight before removing ascites minus weight after removing ascites to indirectly measure the amount of ascites. When we remove the ascites, we need to abdominal aortic blood, centrifuge testing renin, angiotensin II, aldosterone, antidiuretic hormone and other indicators.

RESULTThe effect of Gansui Banixa Tang on increasing the net weight, lowering abdominal circumference and body weight ratio, lowering renin, angiotensin, aldosterone, antidiuretic hormone is better than the other treatment group.

CONCLUSIONIn diuresis party, the group of Gansui Banxia Tang is better than the group of Gansui Banxia Tang remove Zhigancao or Cugansui or Zhigancao and Cugansui, renin-angiotensin-aldosterone system may play a diuretic effect of its one way.

Aldosterone ; metabolism ; Angiotensin II ; metabolism ; Animals ; Ascites ; drug therapy ; metabolism ; physiopathology ; Body Weight ; drug effects ; Diuretics ; pharmacology ; therapeutic use ; Dose-Response Relationship, Drug ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Renin-Angiotensin System ; drug effects

OBJECTIVETo evaluate the inflicted by radar electromagnetic radiation to the sperm DNA of radar operators.

METHODSSperm concentration, viability, motility, sperm abnormality were determined by routine sperm analysis and sperm chromatin structure assay (SCSA) in the highly exposed group(n = 88), lowly exposed group(n = 143) and control group(n = 39).

RESULTSSperm motility, viability of the highly exposed group reduced compared with that of the lowly exposed group and control group, while sperm abnormality increased. The COMP alpha reduction of the highly exposed group indicated that the highly exposed group had a medium fertility potential. The multifactor variable analysis showed that daily working time was a dangerous factor in sperm abnormality and abstinence time was a dangerous factor in the parameter of SCSA.

CONCLUSIONSRadar radiation inflicts damage to male reproduction system and it is important to take protective measures.

Adolescent ; Adult ; Chromatin ; ultrastructure ; DNA Damage ; Dose-Response Relationship, Radiation ; Electromagnetic Phenomena ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Radar ; Spermatozoa ; radiation effects ; ultrastructure

OBJECTIVETo compare the apical microleakage of Vitapex (calcium hydroxide based paste) with that of AH-plus and zinc oxide eugenol sealer when used with laterally condensed gutta percha obturation technique.

METHODSOne hundred single rooted human anterior teeth were instrumented and randomly divided into three experimental groups (A, B, C) of 30 teeth each and two control groups (D, E) of 5 teeth each. Group A was filled with laterally condensed gutta-percha using Vitapex as sealer. Group B was filled with laterally condensed gutta-percha using AH-plus as sealer. Group C was filled with laterally condensed gutta-percha using zinc oxide eugenol as sealer. Group D was the positive control. Group E was the negative control, which were coated with nail polish to entire root surface. Teeth were then suspended in 2% methylene blue. After this, teeth were demineralized dehydrated and cleared. Linear dye penetration was determined under stereomicroscope with calibrated eye piece.

RESULTSThe mean dye penetration for group A, B, C were respectively (0.57 +/- 0.56) mm, (0.79 +/- 0.96) mm and (1.07 +/- 1.12) mm. Group D demonstrated maximum dye penetration. Group E showed no dye penetration. There was no statistically significant difference between group B and group C (P > 0.05). However, there was statistically significant difference between group A and group B, C (P < 0.01).

CONCLUSIONThis study showed that Vitapex used as endodontic sealer material are better than AH-plus sealer and zinc oxide eugenol sealer.

Calcium Hydroxide ; Dental Leakage ; Dental Pulp Cavity ; Gutta-Percha ; Humans ; Molar ; Root Canal Filling Materials ; Root Canal Obturation ; Silicones ; Zinc Oxide-Eugenol Cement

A Kinase Anchor Proteins ; metabolism ; Animals ; Cell Cycle Proteins ; metabolism ; Lipopolysaccharides ; pharmacology ; Liver ; metabolism ; pathology ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVETo investigate effect of tumor necrosis factor-alpha (TNF-alpha) on the Src-suppressed C kinase substrate (SSeCKS) in C6 glioma cells.

METHODSCultured C6 glioma cells were randomly divided into two groups. In time-dependent group, cells were cultured with TNF-alpha (2 ng/mL) for 0 h, 1 h, 3 h, 6 h, 12 or 24 h, respectively; in dose-dependent group, cells were cultured with TNF-alpha (0 ng/mL, 0.02 ng/mL, 0.2 ng/mL, or 2 ng/mL) for 6 h. The expression of SSeCKS was detected by Realtime PCR and Western blot analysis, and immunocytochemistry was used to investigate SSeCKS's subcellular localization.

RESULTSTNF-alpha induced rapid phosphorylations of protein kinase C (PKC) substrates in C6 glioma cells, and upregulated SSeCKS expression in a time and concentration dependent manner. Immunocytochemistry suggested that SSeCKS was localized in the cyroplasm and the leading end of podosomal extensions in control groups, while TNF-alpha induced translocation of SSeCKS perinuclear. This effect could be partly reversed by PKC inhibitor Ro-31-8220.

CONCLUSIONTNF-alpha activates PKC and upregulates SSeCKS expression in C6 glioma cells. These effects are associated with PKC activity, suggesting that SSeCKS plays a role in response to glia activation in PKC mediated pathway.

A Kinase Anchor Proteins ; Animals ; Astrocytes ; metabolism ; Brain Neoplasms ; metabolism ; Cell Cycle Proteins ; metabolism ; Dose-Response Relationship, Drug ; Enzyme Activation ; Gene Expression Regulation ; Glioma ; metabolism ; Immunohistochemistry ; Protein Kinase C ; metabolism ; Protein Transport ; physiology ; Random Allocation ; Rats ; Signal Transduction ; physiology ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; administration & dosage ; physiology

OBJECTIVETo assess the value of monitoring serum human chorionic gonadotropin beta-subunit (beta-HCG) level changes in early diagnosis of ectopic pregnancy.

METHODSTotalling 106 cases of ectopic pregnancy and 69 cases of threatened abortion were included in the study. Blood samples were collected to examine beta-HCG levels, which was repeated 2-3 days later.

RESULTSInitial serum beta-HCG levels of the women with ectopic pregnancy and different were significantly lower than those in women with threatened abortion (P<0.05), and the increment rate of serum beta-HCG in the former was also significantly lower (P<0.05).

CONCLUSIONCompared with single-time serum beta-HCG detection, dynamic monitoring of serum beta-HCG provides more important and reliable evidence for early diagnosis of ectopic pregnancy. Transvaginal ultrasonography and in should also be considered in the clinical analysis.

Adult ; Chorionic Gonadotropin, beta Subunit, Human ; blood ; Early Diagnosis ; Female ; Humans ; Monitoring, Physiologic ; methods ; Pregnancy ; Pregnancy, Ectopic ; blood ; diagnosis ; diagnostic imaging ; Reproducibility of Results ; Sensitivity and Specificity ; Ultrasonography

Adolescent ; Adult ; Aged ; Cord Blood Stem Cell Transplantation ; Female ; Humans ; Liver Cirrhosis ; surgery ; Male ; Middle Aged ; Young Adult

OBJECTIVETo establish a primary culture method of rat testis Leydig cell.

METHODSThe primary rat Leydig cells were treated with or without 4 U/ml human chorionic gonadotropin (hCG), and testosterone in culture medium was detected by radioimmunoassay. The morphology and biological characteristics of Leydig cell were observed.

RESULTSThe culture cells were highly homogeneous, proliferative and had a high differentiation rate. The high purified Leydig cells were verified by their dynamic morphological changes and 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD) histochemical staining. The testosterone secretion induced by hCG significantly increased (P < 0.05) 24 hours after inoculation than that induced without hCG in the control.

CONCLUSIONIt suggests that the Leydig cell cultured in vitro may secrete high concentration of testosterone, and this study laid the basis of androgen replacement therapy for partial androgen deficiency in aging male.

3-Hydroxysteroid Dehydrogenases ; Animals ; Cell Culture Techniques ; Cells, Cultured ; Chorionic Gonadotropin ; pharmacology ; Humans ; Leydig Cells ; cytology ; drug effects ; secretion ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; cytology ; drug effects ; Testosterone ; metabolism