Objective To investigate 60Co γ-ray induced damage in lymphocytes and the relationship between doses of 60Co γ-ray irradiation and the levels of phosphorylated H2AX and ATM.Methods Cells were irradiated with 60Co γ-rays in the range of 0-8 Gy.The levels of phosphorylated H2AX and ATM were detected by Western blot and FACScan,respectively.The micronucleus(MN)was analyzed by CB method to evaluate DNA damage.Results FACScan results showed the dose-effect relationship of γ-H2AX expression were linear.square at 0.5 h post-irradiation to different doses,and the fitting curve was shown as Y=3.96+11.29D-0.45D2.The level of phosphorylated ATM(p-ATM)was not changed significantly by using the same method.Western blot showed that p-ATM protein expression was significandy increased after irradiation compared with sham.irradiated group.The MN assay which represented DNA damage was sensitive to different doses.Conclusions γ-ray irradiation could induce the phosphorylation of H2AX and ATM,which may play an important role in indicating DNA damage.Both of H2AX and ATM have the potential as sensitive biomarker and biodosimeter for radiation damage.

Objective To evaluate the immune response triggered by an in-house constructed hu-man metapneumovirus multi-epitope antigen ( MEA) in a mouse model .Methods Female SPF BALB/c mice at age 4-6 weeks were used in the study and divided into 7 groups.Mice in the five groups including MEA+oligodeoxynucleotides containing CpG motifs ( CpG ODN) intraperitoneal injection ( i.p.) treatment group, MEA+Alum i.p.treatment group, MEA+Alum+CpG ODN i.p.treatment group, MEA+CpG ODN intranasal (i.n.) treatment group and MEA+Alum+CpG ODN i.n.treatment group were immunized three times on days 0, 14 and 21, and those in the other experimental group were immunized intramuscularly with MEA+Quickantibody5W on days 0 and 21.A control group without treatment was set up accordingly .All mice were sacrificed two weeks after the last immunization .Antibodies including IgG , IgG1, IgG2a and IgA in serum samples were detected by ELISA .MTS assay was performed to analyze the proliferation of lympho-cytes.The cytotoxicity of cytotoxic T lymphocytes (CTL) was measured by LDH assay.Flow cytometry was used to detect T lymphocyte subsets .The cytokines secreted by T helper cells ( Th1 and Th2) were analyzed with Bio-Rad Liquid Chips.Results High titers of IgG, IgG1 and IgG2a antibodies were produced in MEA treated mice except for those in intranasal treatment groups .Serum samples from three groups including the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups were positive for IgA antibody .The highest titer of IgA antibody was detected in mice from the MEA+Alum+CpG ODN i.p.treatment group, which was 2.15×103.Compared with the control group, significantly enhanced proliferation of lymphocytes was observed in the MEA+Alum i.p., MEA+Alum+CpG ODN i.p.and MEA+Quickantibody5W i.m.treatment groups (P<0.05).Enhanced cytotoxic activities of CTL were observed in mice with ip.and i.m.treatments as compared with those in control group (P<0.05).The levels of CD4+/CD8+T cells were slightly increased in mice from the MEA+CpG ODN i.p., MEA+Alum+CpG ODN i.p. and MEA+Quickantibody5W i.m.treatment groups as compared with those in control group (P<0.05).In-creased secretion of IL-2, IFN-γand Th2-type cytokines including IL-4, IL-5 and IL-10 were detected in mice from the MEA+CpG ODN i.p.treatment group.The MEA+Alum i.p.treated mice showed a slightly increased secretion of IFN-γand significantly increased secretions of IL-4, IL-5 and IL-10.Significantly in-creased secretions of IFN-γ, IL-4, IL-5 and IL-10 were detected in mice from the MEA+Alum+CpG ODN i.p.treatment group.Significantly increased secretions of IFN-γ, IL-5, IL-10 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were detected in mice from the MEA+Quickantibody5W i.m.treatment group.Conclusion MEA together with different adjuvants could stimulate high titers of specific antibodies , increase the proliferation of lymphocytes and enhance the cytotoxic activities of CTL .CpG ODN could bal-ance the Th1/Th2-mediated immune responses , and the balance could be enhanced when using CpG ODN in combination with Alum .A similar effect could be achieved by using the commercial adjuvant Quickanti -body5w.This study has paved the way for further investigation on the development of hMPV epitope vaccines and diagnostic reagents for hMPV as well as the epidemiological study of hMPV .

Objective To identify the differentially expressed microRNAs in the early transformed cells,the late transformed cells and their parental BEP2D cells.Methods The differentially expressed microRNAs in the above cells were identified by microRNAs microarray assay.Results There were 38differentially expressed microRNAs in R15H20 cells versus BEP2D cells,with 18 upregulated and 20downregulated microRNAs.R15H20 and RHT35 cells shared 25 differentially expressed microRNAs compared with BEP2D cells,with 15 down-regulated and 10 up-regulated microRNAs.There were 87differentially expressed microRNAs in RHT35 cells versus BEP2D cells,with 47 upregulated and 40 downregulated microRNAs.There were 38 differentially expressed microRNAs in RHT35 cells versus R15H20 cells with 20 upregulated and 18 downregulated microRNAs.Conclusions microRNAs are differentially expressed in the different stages of carcinogenesis of BEP2D cells induced by α particles,which suggests that microRNAs may play an important role in α particle-induced malignant transformation of BEP2D cells.

Objective To investigate the effects of 60Co γ-ray partial radiation on chromosome aberration in human peripheral blood in vitro.Methods The samples of heparinized peripheral whole blood from 3 healthy persons were exposed to 60Co γ-rays at the doses between 0 and 8 Gy with the dose rate of 0.35 Gy/min at the temperature of 37 ℃ ,and then mixed with the unirradiated blood samples of the Microscopy was used to observe the chromosome aberration double ( centromere + centromere) and the biological dose was estimated thereby.ResultsThe amounts of double centromere + centromere were increased along with the dose of irradiation in all groups.The estimated biological dose was higher than the 1/3 of the irradiation dose when the dose was between 0.5 to 2 Gy,and was close to the 1/3 of the irradiation dose when the dose was between 4 to 8 Gy.Conclusion Chromosome aberration can be used as a biomarker in estimation of uneven irradiation.

Objective To explore the effect of radon and its progeny on the expression and mutations of p53 in lung tissue of mouse model. Methods Apoptosis was detected by terminal deoxynucleotidy transferase-mediated dUTP-biotin nick end labeling. The expression of p53 gene was analyzed by immunohistochemistry, Western blot and realtime-PCR. PCR-SSCP was used to detect the mutation of p53 in lung tissues. Results Compared with those in the control group, the apoptotic index were increased significantly in 30 WLM and 60 WLM groups( t = 18.11, -10.30,P ＜ 0.05 ). The p53 protein was increased significantly ( t = -11.08, P ＜ 0.05; t = - 7.00, P ＜ 0.0 ) ) in 30 WLM and 60 WLM groups. The mutation of p53 gene was not detected in lungs of radon-exposure mice. Conclusions Lung and bronchus might be the targets of radon and its progeny, and p53 gene plays an important role in the progression of radon-induced lung injury.

Objective To analyze the chromosome aberrations induced by partial radiation in human peripheral blood lymphocytes in vitro.Methods Heparinized whole blood samples were exposed to 2 Gy ~(60)Co 7-rays at 37℃ ,and then mixed with non-irradiated blood by different ratio.The slides were prepared after culturing and the unstable aberrations were analyzed.Results The chromosome aberrations had a good relationship with the ratio of irradiated blood.The chromosome aberrations in partial irradiated group were higher than that in the irradiated group.The estimated dose was 1.27 Gy when the ratio was 1 : 1 ,greater than the dose of 1 Gy.The estimated dose was 0.93 Gy when the ratio was 0.5＝1,also greater than 0.5 Gy.But when the ratio was 1:0,the radiation dose was accordant with the estimated dose.Conclusions Chromosome aberrations could be a biomarker for estimating the uneven irradiation.

The patient,a 11-year-old boy,presented with a 4-year history of erythema and vesicles on the face and arms as well as a 4-month history of tumor and ulcer on the extremities,accompanied by progressive fatigue and intermittent fever.The patient had a body temperature of 37.7℃.No lymph node involvement was observed.Cutaneous examination revealed minimally indurated pink-red patches on the face and nose and dusky red firm nodules and tumors of varying sizes on the extremities.The nodules ranged from 2.0 cm to 18 cm in diameter,some had necrosis and black crusts on the surface.Ulcers were observed in some of the larger nodules;many of the ulcers extended into the muscle layer.White purulent discharge was seen on the surface of many of the nodules.The lesions were sharply demarcated,firm,tender, and surrounded by small satelite nodules.Histologically,there were large quantities of irregularly shaped, middle-sized tumor cells with clear cytoplasm,large twisted nuclei and prominent chromatin,infiltrating from the epidermis to subcutaneous tissue.The tumor cells infiltrating the follicles and eccrine sweat glands were either distributed perivascularly in a nest shape,or dispersed.There were broken nuclei and reactive histio- cytic infiltration in the dermis and subcutaneous tissue.Immunohistologically,the tumor cells were positive for cytoplasmic CD3 around the nuclei,for CD56,CD45RO and T cell intracellular antigen-l,and partly for CD30,CD8 and Ki67.Epstein-Barr virus-encoded nuclear RNA was positive with in situ hybridization. TCR?-2 gene rearrangement was positive in these tumor cells.A diagnosis of hydroa vacciniforme-like primary cutaneous NK/T-cell lymphoma was made.Therefore,this is a case report of hydroa vaccini- forme-like primary cutaneous NK/T-cell lymphoma with primary involvement in the skin;the condition was slowly progressive over 51 months.

AIM: To observe the expression of β-catenin in patients with chronic myeloid leukemia (CML) at different disease phases, and to analyze the relationship between BCR-ABL and cytogenetic response to imatinib mesylate. METHODS: RT-PCR and Western blotting were used to detect β-catenin mRNA and protein expression in bone marrow mononuclear cells (BMMNCs) from 99 patients with CML. The association with BCR-ABL and BCR-ABL fusion was determined by FISH in 94 patients after one year treatment with imatinib mesylate, and the relationship between β-catenin and cytogenetic response to imatinib mesylate was analyzed. RESULTS: The expression of β-catenin was increased significantly in patients with blast crisis and accelerated phase (P<0.01), while the expression of β-catenin between normal person and chronic phase of CML patients was not statistically different (P>0.05). No significant relation between β-catenin and BCR-ABL expression (r=0.314, P>0.05) was observed. The expression of β-catenin was increased significantly in the patients who did not reach main cytogenetic remission (P<0.01). CONCLUSION: The patients in progression phases of CML over-express β-catenin. The expression of β-catenin is not significantly related to BCR-ABL expression, but related to the therapeutic response of imatinib. Beta-catenin may be involved in the mechanism of CML progression and could be used as a new therapeutic target.

Effect of strophanthidin (Str) on intracellular calcium concentration ([Ca2+]i) was investigated on isolated ventricular myocytes of guinea pig. Single ventricular myocytes were obtained by enzymatic dissociation technique. Fluorescent signal of [Ca2+]i was detected with confocal microscopy after incubation of cardiomycytes in Tyrode's solution with Fluo3-AM. The result showed that Str increased [Ca2+]i in a concentration-dependent manner. The ventricular myocytes began to round-up into a contracture state once the peak level of [Ca2+]i was achieved in the presence of Str (10 μmol·L-1), but remained no change in the presence of Str (1 and 100 nmol·L-1). Tetrodotoxin (TTX), nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str (1 and 100 nmol·L-1), but had no obvious effects on the action of Str (10 μmol·L-1). The elevation of [Ca2+]i caused by Str at all of the detected concentrations was partially antagonized by rynodine (10 μmol·L-1) or the removal of Ca2+ from Tyrode's solution. In Na+, K+-free Tyrode's solution, the response of cardiomycytes in [Ca2+]i elevation to Str (10 μmol·L-1) was attenuated, while remained no change to Str (1 and 100 nmol·L-1). TTX, nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str at all of the detected concentrations in Na+, K+-free Tyrode's solution. The study suggests that the elevation of [Ca2+]i by Str at the low (nomomolar) concentrations is partially mediated by the extracellular calcium influx through Ca2+ channel or a "slip mode conductance" of TTX sensitive Na+ channel. While the effect of Str at high (micromolar) concentrations was mainly due to the inhibition of Na+, K+-ATPase. Directly triggering the release of intracellular Ca2+ from sarcoplasmic reticulum (SR) by Str may be also involved in the mechanism of [Ca2+]i elevation.

Objective To investigate the indications and results of long channel technique with pedicle inner preputial flap urethroplasty for middle hypospadias repair. Methods During September 2007 to April 2010,28 cases of middle hypospadias were include in this study.The average age was 2.0 years (1.5 -6.0 years).The orifice of urethra was opened in the shaft of penis,and the distance between the urethral meatus and the center of glans was 25 mm ( 16 - 37 mm).A pars-meatus skin incision was made,which was dissected deep to the urethral plate.The urethral plate was transected,and a long channel between the urethral plate and the corpora cavernosa was created.The neourethra was made from the inner prepuce,and transposed to the ventral tunnel through a tunnel between buck's fascia and albuginea of ventrolateral corpora cavernosa.The neourethra was anastomosed with the proximal urethra.The buck's and dartos fascia along the skin incision were brought together and sutured individually,covering the proximal neo-urethra and the anastomosis.Induced penile erection confirmed that 13 cases were with mild penile curvature,and the other 15 case were without penile curvature.The average length of the defected urethra and tunnel was 38 mm (30 -42 mm) and 33 mm (26 -38 mm). ResultsSuccess was achieved in all cases without fistula or urethral stricture formation with the average follow-up of 20 months (6 -31 months),and penile curvature was completely corrected.The urethra was opened in the apex of the glans with normal-looking circumcised penis. ConclusionsLong channel technique with pedicle inner preputial flap urethroplasty can provide another option for repairing middle hypospadias without penile curvature or with mild penile curvature,especially for young children and those with small penis.This technique is simple,and the result is satisfied.