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 evaluate the roles of Toll-like receptor 2 (TLR2),TLR4 and myeloid differentiation factor 88 (MyD88) in immune recognition and immune mediation in sporotrichosis by detecting their expressions in skin lesions of sporotrichosis.Methods Biopsy specimens were obtained from the skin lesions of 19 patients with sporotrichosis and normal skin of 12 healthy human controls.Immunohistochemistry was performed to observe the expressions of TLR4 and MyD88,and real-time fluorescence-based quantitative PCR to quantify the expressions of TLR2 and MyD88 mRNAs.Data were expressed as mean ± standard error.Statistical analysis was done with the SPSS17.0 software.Independent samples t-test was conducted to compare the parameters between the lesional and control specimens.A p-value of less than 0.05 was considered to be statistically significant.Results TLR4 and MyD88 were mainly observed in the whole epidermis except the stratum corneum as well as plasma cells and lymphocytes in the dermis and subcutaneous tissue in lesional skin.However,TLR4 was nearly absent and MyD88 was weakly expressed in the dermis and subcutaneous tissue in normal skin.The expression levels of TLR4 and MyD88 were significantly higher in the lesional skin than in the control skin (TLR4,63.767 ± 3.829 vs.5.167 ± 3.246,t =4.82,P ＜ 0.05; MyD88,57.236 ± 4.744 vs.10.588 ± 1.640,t =3.30,P ＜ 0.05).Similarly,the lesional skin showed significantly stronger expressions of TLR2 and MyD88 mRNAs compared with the normal skin (TLR2,1.974 ± 1.452 vs.1.430 ± 1.073,P ＜ 0.05; MyD88,2.028 ± 2.061 vs.0.688 ± 0.422,P ＜ 0.05).Conclusion Sporothrix may induce the development of sporotrichosis by interacting with host immune system via TLR signaling pathways.

Objective To provide a practicable method for the complete display and localization of the posterolateral structures (PLS) of the normal knee through MRI study. Methods 30 tibial bone specimens were observed to establish the bony landmark for localizing the knee. In 50 cadaver knees, the angles between lateral tibial plateau and the long axis of the individual structure of PLS were measured. Then the scan methods of the oblique MR images were determined based on above results. The routine and oblique scans of T 1WI were performed in 40 normal knees. The display effect and appearance of the PLS were observed on MRI. Results The lateral tibial plateau was a stable bony landmark for measuring and localizing of the knee. In the 40 normal knees, The fibular collateral ligament could be intactly displayed on 70? posterior coronal oblique images in 34 cases (85%). The popliteus could be better seen on either 45? medial sagittal oblique in 34 cases (85%) or 60? posterior coronal oblique planes in 36 cases (90%). The popliteofibular ligament could be intactly appreciated on both 60? posterior coronal oblique in 32 cases (80%) and 70? lateral sagittal oblique images in 34 cases (85%). Although the arcuate ligament and the fabellofibular ligament could occasionally be seen on routine and oblique images, but the display rate was lower. Conclusion The oblique MR imaging can intactly display the main structures of PLS, and can be useful in diagnosing the injuries in those structures.