Objective To investigate the effects of staphylococcal enterotoxin A SEA gene on target cells mediated by replicative-deficient recombinant adenovirus vector. Methods Lymphocytes of C57BL/6 mice were infected with various titers of recombinant adenoviruses. Supernatants were collected after 12 h, 24 h, 48 h, 72 h, 96 h, 120 h and 144 h of incubation and analyzed for proliferation of lymphocytes by MTT assay. IL-2 level in the culture supernatants was measured with ELISA. The killing effect of lymphocytes was also observed by MTT assay. Results Proliferation response and elevated levels of IL-2 were observed in experimental group. The killing effect on B16 cells was stronger in experimental group, which seemed to be dose-dependent with the increase of ratio of lymphocytes/target cells. Conclusions SEA gene can be expressed in lymphocytes of C57BL/6 mice mediated by replicative-deficient recombinant adenovirus vector. The expressing products can activate lymphocytes of C57BL/6 mice, which kill B16 cells in vitro.

Objective To probe into the application of left atrial volume tracking technique(LAVT)on the evaluation of right atrial function in patients with pulmonary hypertension.Methods Forty-one patients with pulmonary hypertension (pulmonary hypertension group) and 37 control subjects (control group) were involved.Right atrial maximal volume (RAVmax),right atrial presystolic volume(RAVpre),right atrial minimal volume (RAVmax),systolic right atrial filling rate (dv/dtS),early diastolic right atrial emptying rate(dv/dtE) and late diastolic right atrial emptying rate(dv/dtA) was derived by LAVT.Right atrial passive emptying volume (RAVp),right atrial passive emptying fraction (RAVpEF),right atrial active emptying volume (RAVa),right atrial active emptying fraction (RAVaEF),right atrial total emptying volume (RAVt)and right atrial total emptying fraction (RAVtEF) was calculated.All the right atrial volume parameter was corrected by body surface area to obtain right atrial volume index (RAVI).Results RAVImax,RAVImin,RAVIpre,RAVIt,RAVIa,dv/dtS and dv/dtA in pulmonary hypertension group was higher than that in control group [(78.39 ± 49.35) ml/m2 vs.(24.80 ± 11.91) ml/m2,(62.59 ± 46.56) ml/m2vs.(17.46 ± 8.40)ml/m2,(70.12 ± 48.03) ml/m2 vs.(20.02 ± 9.46) ml/m2,(18.77 ± 11.47) ml/m2 vs.(9.35 ± 6.74) ml/m2,(8.53 ± 9.81) ml/m2 vs.(3.25 ± 3.00) ml/m2,(145.85 ± 80.56) ml/s vs.(86.44 ± 48.46) ml/s,(155.63 ±126.47) ml/s vs.(67.74 ± 33.27) ml/s],and RAVIp in pulmonary hypertension group was lower than that in control group [(6.09 ± 5.16) ml/m2 vs.(10.23 ± 11.12) ml/m2],and there were significant differences (P ＜0.05).But there were no significant differences in RAVItEF,RAVIpEF,RAVIaEF and dv/dtE between two groups (P＞0.05).Conclusions In patients with pulmonary hypertension,right atrial booster pump function and reservoir function increases,while right atrial conduit function decreases.LAVT has a potential ability to evaluate right atrial function.

Recepteur d′origine Nantais(RON),a tyrosine kinase receptor ,is a growth factor receptor belonging to the proto-oncogene met family and has been proved to display abnormal expres?sion in many types of tumors. The RON receptor is activated by binding to the ligand macrophage stim?ulating protein,overexpression of the receptor,variants of the proto-oncogene and by point mutations of the kinase region. The downstream transduction of RON by mitogen-activated protein kinases and phosphoinositide3-kinase signaling pathways can help regulate the proliferation,apoptosis,migration and invasion of tumor cells. A better understanding of the mechanisms and related signaling pathways of RON activation in tumor progress and development will provide more information for the RON-based target therapy.

T mutation was detected in the 4th propositus at the 9th intron,but any COL1A1 or COL1A2 gene mutation was detected in the third propositus and the other members in the former families.Conclusions The genetic mutation of COL1A1 may result in OI in China,but other mutations may also exist.Moreover,the phenotype was influenced not only by OI genotype,but also by the genetic background,environment and other factors.

Objective　To analyze the effects of flexible ureteroscopic Holmium laser lithotripsy on renal function in the treatmen of renal calculi.　Methods　From October 2010 to March 2012,41 cases of solitary renal calculi (24 males and 17 females) were treated with flexible ureteroscopic Holmium laser lithotripsy ( FURL).Patient's mean age was 51.9 ± 15.9 years ( range from 31 to 88 years).Locations of renal calculi detected by image study were 22 cases in middle and upper calyx,9 cases in lower calyx,10 case in renal pelvis.The mean size of calculi was 16.9 ±6.0 mm (range from 10 to 28 mm).Blood samples (2 h pre-operatively,and then 2 h,12 h,24 h,48 h,72 h post-operatively) were collected and serum　NGAL,serum Cys-C were tested　Results　The measured levels of pre-operative NGAL and Cys-C were 3.5 ± 0.6μg/L,501.7±121.3 μg/L,and levels of post-operative NGAL at 2 h,12 h,24 h,48 h and 72 h were 4.2±0.8 μg/L,5.0±1.0 μg/L,4.9±1.4 μg/L,4.3± 1.1 μg/L and 3.8 ±0.1 μg/L,while the according levels of Cys-C were ( 516.4 ± 126.2 ) μg/L,( 723.8 ± 134.8 ) μg/L,( 770.4 ± 162.8 ) μg/L,(671.7 ± 138.3 ) μg/L and 574.0 ± 116.7 μg/L.Serum NGAL began to increase 2 hours after operation (P ＜0.05,and reached the peak in 12 hours,it began to decline 12 hours after surgery,but it was still higher at 72 hours than pre-operative level (P ＜ 0.05.Serum Cys-C showed no obvious ascent 2 hours after operation in FURL group ( P ＞ 0.05 ),but increased obviously 12 hours after operation and lowered down after the peak that occurred 24 hours after surgery.Serum Cys-C still remain above the baseline 72 hours after operation (P ＜0.05).　Conclusions　Flexible ureteroscope lithotripsy can cause reversible damage to renal function after surgery.

Objective:To study the inhibitory effect of siRNA on Heparanase (HPA) expression in SKOV3 cells. Methods:Two pairs of 21 bp reverse repeated sequence targeting HPA RNA (spaced by 9 bp nueleotide) were synthe- sized and were cloned into plasmid PGenesil-1 to construct recombinant plasmid PGenesil-1(+)-HPA expressing 2 hair- pin siRNAs.The inhibition of HPA gene was detected by RT-PCR,real-time PCR and immunohistochemical staining after PGenesil-1(+)-HPA was stably transfected into SKOV3 cells.Results:The recombinant plasmid PGenesil-1(+)-HPA (expressing 2 hairpin siRNAs) was successfully constructed.RT-PCR,real-time PCR and immunohistochemical staining showed that there was a significant decrease in HPA mRNA and protein level in experimental group compared with those in control group.Conclusion:siRNA targeting HPA mRNA can specically suppress the expression of HPA gene in SKOV3 cells;RNA interference method provides a new way for studying the role of HPA and gene therapy of cancer.

ObjectiveTo explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group(irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group(treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.ResultsThe mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P ＜ 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P ＜ 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P ＞ 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P ＞ 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P ＜ 0.05).ConclusionsHeat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.

ObjectiveTo investigate the effect of heat treatment combined with narrow band ultraviolet B(NB-UVB) on cultured normal human melanocytes in vitro.MethodsMelanocytes were isolated from the foreskin of normal human,cullured in vitro,and irradiated with NB-UVB of different doses(20,30,50,70,90,120 and 180 mJ/cm2).Then,MTT assay was performed to evaluate the proliferation and activity of melanocytes to determine the optimal dose of UVB for the next experiment.Melanocytes were classified into 3 groups to be treated with heat at 42 ℃ for 1 hour (heat group),irradiated with UVB at 50 mJ/cm2 (UVB group),or irradiated with UVB at 50 mJ/cm2 followed by heat treatment at 42 ℃ for 1 hour (combination group),daily for 3 successive days; those receiving no treatment served as the control.After 24-hour culture following the last treatment,tyrosinase activity was evaluated with L-dopa as the substrate,melanin content was detected by NaOH assay,and cell cycle stages were determined by flow cytometry.ResultsNB-UVB irradiation decreased the viability of melanocytes in a dose-dependent manner,and the optimum dose of UVB was 50 mJ/cm2.The tyrosinase activity of melanocytes was 0.244 ± 0.018 and 0.310 ± 0.015 respectively in the UVB group and combination group,and increased by 3.8％ (P ＜ 0.05) and 31.9％ (P ＜ 0.05) respectively compared with the control group (0.235 ± 0.018); the melanin content was 0.201 ± 0.016 and 0.286 ± 0.019,respectively in the UVB group and combination group,and increased by 17.5％ (P ＜ 0.05 ) and 67.3％ (P ＜ 0.05) compared with the control group (0.171 ± 0.016).In comparison with the control group,the percentage of melanocytes in G1 phase was decreased by 23.94％ in the UVB group(P＜ 0.05) and 33.51％ in the combination group(P ＜ 0.05),while that in S phase and G2 phase increased by 15.35％ (P ＜ 0.05 ) and 11.93％ (P ＜ 0.05),respectively in the UVB group,and 17.76％ (P ＞ 0.05) and 16.08％ (P ＞ 0.05),respectively in the heat group.ConclusionHeat treatment and NB-UVB can synergistically enhance the tyrosinase activity and accelerate melanogenesis,proliferation and differentiation,of melanocytes.

Objective To explore the expression and function of miR-125b, miR-30b and miR-424 in endometrial cells. Methods Human endometrial samples were obtained in natural cycles and stimulating cycles. Endometrial epithelial cells (EECs) and endometrial stromal cells (ESCs) were isolated and confirmed by immunofluorescence. The expressions of miR-125b, miR-30b and miR-424 were detected by real-time PCR. Results The expression levels of miR-125b, miR-30b and miR-424 were higher in proliferative phase in ESCs than those in EECs. And in EECs, the expression levels of miR-125b, miR-30b and miR-424 were significantly up-regulated in secretory phase than in proliferative phase, while it was stable in ESCs. In addition, the expressions of miR-125b in EECs and miR-30b were increased in ESCs in women with elevated progesterone on the day of HCG administration than those of the control. The target genes of miR-125b, miR-30b and miR-424 mainly participated in cell migration and motion, cell-cell adherens junction and Wnt signaling pathway. Conclusion miR-125b, miR-30b and miR-424 were differently expressed in endometrial cells in different phases, and may participate in regulation of endometrial receptivity.

Objective To evaluate the effect of heparin binding epidermal growth factor-like growth factor (HB-EGF)on liver regeneration after partial orthotopic liver transplantation.Methods Fourty SD rats were used to establish the model of partial orthotopic liver transplantation with ameliorated two-cuff technique.Then all the rats were divided into 2 groups:experiment group and control group.Twenty rats of experiment group were adminis- tered 500?g/kg HB-EGF via vena caudalis immediately after operation twice a day,while the same volume of saline was administered to the rats in control group.Five rats in each group were selected randomly and killed at the 6th hour,day 2,4 and 7 after operation,respectively.The serum levels of albumin(Alb)and alanine aminotransferase (ALT)in the blood sample were detected.Every liver was removed and weighed.The expression of Ki-67 was de- tected by using immunohistochemistry assay.The regeneration activity of hepatocytes was evaluated by flow cytom- etry.Results The wet weights of liver in experiment group were all significantly higher than that in control group at the 6th hour,day 2 and 4 after transplantation(P