Objective To evaluate the growth inhibition and apoptosis of human monocytic leukemia THP-1 cell line by using 5,8-dimethyl-2-β-hydroxyisovalerylshikonin (SK36) and explore its preliminary mechanism. Methods CCK colorimetric assay and cell counting was used to examine the growth inhibition of shikonin on THP-1 cells. The apoptosis of THP-1 cells was detected by Annexin V/PI double labeling. The activation of Caspase-3 apoptosis pathway was determined by FCM. The apoptosis and the necrosis of THP-1 cells were detected by the laser scanning confocal microscopy. Results When the THP-1 cells were treated with SK36 at 1.02 μg/ml for 24 h and 48 h, the growth inhibition was dose-dependent. The cell apoptotic rate of THP-1 cells treated with 1.02 μg/ml evaluated by FCM with Annexin V/PI double labeling staining were (40.61 ±2.13) % and (67.40±9.15) % at 24 h and 48 h after treatment, respectively, which were significantly higher than that of the control group [(16.97±0.61) %] ([ = 18.444, t = 9.528, P <0.01). SK36 could induce THP-1 cells apoptosis involving the activation of Caspase-3 (F= 323.61, P<0.01). Conclusion SK36 can induce human THP-1 cells to undergo apoptosis, and its primary mechanism was to activate the Caspase-3.

Objective To compare the two ways of twice irradiation on the rabbit liver VX2 tumor using high intensity focused ultrasound (HIFU).Methods Totally 45 tumor-bearing rabbits were randomly divided into 3 groups:group A(one-time irradiation group) received a one-time ablation;group B and C(twice irradiation group) firstly received a low-dose irradiation(without ablation),then group B received ablation on the next day while group C received it 2 days later.Results Compared with one-time irradiation group,the total treatment time of twice irradiation group was not significantly different,but the time of each irradiation,the incidence of skin erythema and the energy efficiency factor(EEF)were less,and that of group B were the least.After ablation,the typical coagulation necrosis in tumor tissues occurred,and the recurrence and metastasis were effectively controlled in all the three groups.Conclusion The total treatment time and efficacy of twice irradiation were same as one-time ablation,but the time and dose of each irradiation significantly decreased,the damage efficiency was enhanced and the complications were reduced.The way of continuous two-days twice irradiation was the most effective,which would be a safe and effective method of HIFU treatment.

Objective To investigate the change of peripheral blooe T cell subsets,NK cells ane serum soluble interleukin-2 receptor(sIL-2R)concentration in patients with gastric cancer before ane after surgery,ane to uneerstane immune function status ane changes of perioperative gastric cancer patients. Methods One huneree ane thirty-five perioperative gastric cancer patients were selectee as our subjects who hospitalizee from May 2009 to May 2011 in Tumor Hospital of Shanxi Province,ane they were servee as treatment group,while 50 healthy subjects were selectee as controls. The number of CD3 + ,CD4 + ,CD8 + T cells,rate of CD4 + / CD8 +ane the NK cell ratio in blooe cells were eetectee by flow cytometry. ELIAS was appliee to measure serum sIL-2R concentration. Observee the above ineexes of control group at the same perioe,ane comparee the ineexes before operation of 1 e of the treatment group. Results The rate of NK cells,CD3 + T cells ane CD4 + / CD8 +ratio in patients at pre-operation were(10. 11 ± 3. 64)% ,(55. 60 ± 9. 61)% ,(30. 22 ± 6. 17)% ,1. 14 ± 0. 35,respectively,lower than that of control group(( 28. 39 ± 5. 81 )% ,( 68. 65 ± 7. 39 )% ,( 47. 87 ± 4. 85)% ,1. 82 ± 0. 24 respectively;t = - 5. 9,8. 6,8. 2,12. 7;P < 0. 01). CD8 + T positive rate of cells increasee from(27. 05 ± 7. 86)% to(34. 26 ± 6. 23)%(t = - 6. 5,P < 0. 01). At 14th eay after surgery,the cell immune function of the patients recoveree graeually,ane there were statistically significant eifferences in the above ineexes comparee with pre-operation(P < 0. 05). The eramatic changes were seen among patients with the late Gastric cancer TNM staging. Comparee with patients with stage Ⅳ,all above ineex were significant eifferent from that of patients with stageⅠ,Ⅱ(P < 0. 05),ane no significant eifferences was seen in patients with stageⅢ(P > 0. 05). The concentration of serum sIL-2R in patients with gastric cancer before operation was(575. 71 ± 34. 77)U/ L,higher than that of healthy persons((428. 26 ± 21. 77)U/ L,t = - 7. 9,P < 0. 01),ane serum sIL-2R levels in patients with stage Ⅲ,Ⅳ was lower than that of patients with stage Ⅰ,Ⅱ patients with low(P< 0. 05). Conclusion The immune function of patients with gastric carcinoma is relatee to tumor loae size ane eifferent pathological staging. The ineex of the ratio of NK cells ane T lymphocyte subsets,serum sIL-2R levels can be servee as ineicators for monitoring perioperative evaluation of prognosis of gastric cancer.

Objective To investigate the changes of CD4 +CD25highFoxp3 +regulatory T (Treg) cells and their significance in immune escape of childhood B-cell acute lymphocytic leukemia ( B-ALL ) . Methods Forty-two children with B-ALL and twenty-eight age-matched healthy children were enrolled in this study.Flow cytometry analysis was performed to evaluate the proportion of CD 4 +CD25high Foxp3 +Treg cells as well as CD4 +CD25high ICOS+Foxp3 +and CD4 +CD25high ICOS-Foxp3 +subsets in peripheral blood samples.The expression of associated molecules including IL-10, TGF-β, IL-35, TGF-βRII, ICOS and CD28 at protein level were also measured by flow cytometry analysis .The transcription level of Smad3/4, TIEG1 and Itch by CD4 +T cells were determined by quantitative real-time PCR.The concentration of TGF-βin plasma was detected by enzyme-linked immunosorbent assay.Results (1)The proportion of CD4 +CD25highFoxp3 +Treg cells in children with B-ALL were significantly higher than those of health subjects (P<0.05).The proportion of both ICOS +Foxp3 +and ICOS -Foxp3 +subsets were increased in comparison with those of control group (P<0.05), while the ratio of ICOS +Foxp3 +to ICOS-Foxp3 +was decreased (0.73 ±0.21 vs 1.87 ±0.59, P<0.05).(2) The expression of Foxp3, TGF-β, IL-10 and IL-35 by ICOS+Foxp3 +Treg cells and the expression of membrane bound TGF-βby ICOS -Foxp3 +Treg cells were significantly increased in children with B-ALL (P<0.05).However, the expression of Foxp3 by ICOS -Foxp3 +Treg cells showed no significant difference between the two groups (P>0.05).(3)The concentra-tion of TGF-βin plasma from children with B-ALL were higher than those from control group [ ( 25 .83 ± 12.65) ng/ml vs (8.59 ±5.73) ng/ml, P<0.05].The expression of TGF-βRII and its associated mole-cules (Smad3/4, TIEG1 and Itch) by CD4 +T cells were significantly up-regulated.Moreover, an increased expression of ICOS and CD28 by CD4 +CD25highFoxp3 +Treg cells were also observed in children with B-ALL (P<0.05).Conclusion The hyper-activity of TGF-β, ICOS and CD28 signaling might be closely associ-ated with the increased proportion of CD4 +CD25high Foxp3 +Treg cells and the imbalance of its subsets in children with B-ALL.

OBJECTIVETo determine the optimal concentration of c-erbB2 antisense probe labeled with superparamagnetic iron oxide (SPIO) nanoparticles for in vivo tumor imaging in mice using magnetic resonance imaging (MRI).

METHODSThirty BALB/c mice bearing SK-Br-3 tumor were randomized into 5 groups to receive injections of different concentrations of SPIO-labeled c-erbB2 antisense probe (containing 6.0, 9.0, 12.0, 15.0, or 18.0 mg Fe/kg). MRI was performed before and 6 h after the injections, and the signal intensities of the tumor were compared among the groups. The tumor tissues were then dissected for microscopic examination with HE and Prussian blue staining.

RESULTSThe tumor-bearing mice all survived after injections of the probe at doses of 6.0, 9.0 and 12.0 mg, but injections at higher doses (15.0 and 18.0 mg) caused death in some mice. Injections of the probe at the doses of 12.0, 15.0 and 18.0 mg resulted in significant signal enhancement of the tumor (P<0.001) to allow visual identification, but the changes showed no significant differences among the 3 groups (P>0.05). Pathological examination revealed irregular structures of the tumor issue containing numerous heterogeneous tumor cells aligned into cancer nests; Prussian blue staining visualized scattered blue iron particles in the tumor issue, which was especially obvious in mice injected with 12.0, 15.0 and 18.0 mg labeled probe.

CONCLUSIONInjection of 12.0 mg/kg SPIO-labeled c-erbB2 antisense probe allows optimal tumor imaging in BALB/c mice using MRI.

Animals ; Antisense Elements (Genetics) ; Cell Line, Tumor ; Contrast Media ; Dextrans ; Genes, erbB-2 ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Nanoparticles ; Nucleic Acid Probes ; genetics ; Xenograft Model Antitumor Assays

Objective To evaluate the safety and immunogenicity of one booster dose of inactivated hepatitis A vaccine in young adults.Methods The subjects were selected from participants in the clinical trial of immunogenicity of inactivated and attenuated live hepatitis A vaccine in young adults.Eligible subjects were those who had received one dose of inactivated or attenuated hepatitis A vaccine,could be contacted and were sero-negative before primary vaccination.All qualified subjects were immunized with one booster dose of inactivated hepatitis A vaccine.The blood samples were collected before booster dose vaccination and 28 days after the immunization.Anti-HAV antibody titer ≥20 mIU/ml was considered to be sero-protected against hepatitis A virus.Results The GMCs in the inactivated HAV vaccine group and attenuated live vaccine group before booster dose vaccination were 70.80 mIU/ml and 50.12 mIU/ml,respectively,and the sero-protection rates were 94.7％ and 65.0％,respectively.After the vaccination of the booster dose,the sero-protection rates in both groups were 100.0％,and the GMCs were 2 816.09 mIU/ml and 2 654.55 mIU/ml,respectively.Conclusion The GMCs and sero-protection rates of anti-HAV antibody in young adults declined after three years of the primary vaccination.However,the higher GMC and sero-protection rate were observed in the inactivated vaccine group than in the attenuated live vaccine group.Significant increases of GMC levels were observed in both groups after one booster dose vaccination.