In order to determine the relationship between pulmonary leukosequestration and oxygen free radical generation, We prospectively studied 20 patients undergoing cardiopulmonary bypass for open-heart procedure of congenital heart discases. Serum levels of SOD and MDA were measured at the begining and the end of extracorporeal circulation cell counts and differential counts were made from blood in the RV and LA after CPB. The results showed that CPB caused significant increase in WBC, Polymorphonuclear cells were the main species of leukocytes sequestered in the lungs. (Median cell difference 2.018?10~9/L, P

ObjectiveTo study the epidemiological characteristics of leprosy in China,2001-2010.MethodsData were collected from the database of the national system of leprosy surveillance.A descriptive and comparative analysis was performed.ResultsTotally,15 507 new leprosy cases were detected from 2001 to 2010 with an average case detection rate of 0.118 per 100 000 population.Among these cases,2.7％ were children under 15 years,86.5％ multibacillary,and 22.5％ suffered from grade 2 disability.From 2001 to 2010,a total of 1506 relapse cases were detected,and relapse occurred after multi-drug therapy(MDT) in 464 of these cases.There were significant differences between low and high endemic areas in the proportion of children under 15 years,females,immigrant patients among newly detected patients as well as the proportion of cases of relapse after MDT among all the relapse cases.By the end of 2010,the registered leprosy cases were 6032 with a prevalence rate of 0.450 per 100 000 population,among whom 2886 were under MDT.ConclusionsThe leprosy case detection rate continued to decrease in China from 2001 to 2010 with an unequal distribution.The pocket areas were in Yunnan province,Guizhou province,Sichuan province,Guangdong province,Hunan province and Tibet Autonomous Region.It is warranted to enhance the control of leprosy and reduce the prevalence of disability due to leprosy.

Objective To select microRNAs (miRNA) associated with human cutaneous malignant melanoma (MM). Methods Total RNA was extracted from 6 tissue samples of MM and 9 human control samples of pigmented nevi, and small RNAs of less than 200 bp were enriched, miRNA microarray was used to select differentially expressed miRNAs between tissue samples of MM and pigmented nevi from 468 candi-dates. The expression of differentially expressed miRNAs was confirmed by fluorescence based real-time quan-titative PCR (qPCR) in all of these samples. Those miRNAs that were identified as differentially expressed with both miRNA microarray and qPCR were considered as significant miRNAs. Results Between the tissue samples of MM and pigmented nevi, 12.18% to 86.33% of miRNAs differentially expressed by more than 2 folds, 1.28% to 19.02% by more than 5 folds, and 0.43% to 5.34% by more than 10 folds. The expression of miRNA-21 was obviously up-regulated, while that of miRNA-320 and miRNA-494 was down-regulated in the MM samples. Conclusion There is an increase in the expression of homo sapiens miRNA-21 but a decrease in that of miRNA-320 and miRNA-494 in MM tissues.

Objective To analyze the epidemiological features of leprosy in China from 2011 to 2015,and to provide scientific evidences for prevention and treatment strategies.Methods An epidemiological analysis and a trend analysis were conducted based on the national leprosy surveillance data from 2011 to 2015.Results The leprosy detection rate in China decreased from 0.085 per 100 000 in 2011 to 0.049 per 100 000 in 2015,with an average annual decline rate being 12.9％.A total of 4 775 leprosy cases were newly detected during 2011-2015,including 106 (2.2％) children,1 499 (31.4％) females,518 (10.8％) floating people,4 041 (84.6％) multibacillary cases and 1 134 (23.7％) cases with grade 2 disabilities.From 2011 to 2015,328 relapsed cases were reported,including 153 (46.6％) cases recurring after combined chemotherapy.The prevalence rate of leprosy in China decreased from 0.407 per 100 000 in 2011 to 0.235 per 100 000 in 2015,with an average annual decline rate being 12.9％.By the end of 2015,there had been 3 230 registered leprosy cases and 124 counties with a prevalence rate above 1 per 100 000.Conclusions The detection rate and prevalence rate of leprosy in China were both decreasing continuously from 2011 to 2015.The high-epidemic provinces were Yunnan,Guizhou,Sichuan and Guangdong.However,leprosy control in middle-and low-epidemic provinces can not be ignored.

Objective To study the changes of plasma ET-1,CGRP,TXA2 and their receptor expression in lung tissue of children with congenital heart disease-assicated pulmonary artery hypertension.Methods Plasma was collected from 60 children patients.ET-1,CGRP and TXB2 levels were measured by EIA,RIA and ELISA,respectively.Expression of their receptors in the lung tissue was detected by RT-PCR.Results The plasma ET-1 and TXB2 levels were significantly higher in CHD patients than in control children.The plasma CGRP level was significantly lower in CHD patients than in control children.Fourteen days after treatment with PGE1,the plasma ET-1 and TXB2 levels were significantly decreased with no significant changes in CGRP.The expression level of ET-1 and TXA2 receptor mRNA was significantly higher in patients with pulmonary artery hypertension than in those without pulmonary artery hypertension.However,the expression of CGRP receptor mRNA was lower in patients with pulmonary artery hypertension than in those without pulmonary artery hypertension.Conclusion Plasma ET-1,CGRP and TXA2 levels are closely related with congenital heart disease-associated pulmonary artery hypertension,and can be used as an index for its prognosis.

Objective: To investigate the killing activity of cytokine-induced killer (CIK) cells after incubation with autologous tumor cell lysate-pulsed dendritic cells (Ag-DC) and to evaluate the immune functions of patients, the clinical efficacy and side effect of Ag-DC in combination with CIK on hepatocellular carcinoma. Methods: Peripheral blood mononuclear cells were isolated from 24 patients with hepatocellular carcinoma, and cultured with granulocyte-macrophage colony-stimulating factor and interleukin-4 to produce dendritic cells (DCs). The DCs were pulsed with autologous tumor cell lysate. T lymphocytes from PBMC were cultured with interferon-Y (IFN-γ), IL-2, CD3-moAb, and IL-1α to prepare CIK. The killing effect of CIK on SMMC-7721 was investigated after CIK was incubated with Ag-DC. After immunotherapy with Ag-DC and CIK, immunolog-ic and clinical responses of the 24 patients were evaluated. Results: The killing effect of CIK was remarkably improved after CIK was incubated with Ag-DC. The immunotherapy with DC and CIK alleviated symptoms and improved the immune functions of the patients. Except for transient fever and chill, no remarkable adverse events were observed. Conclusion: Ag-DC in combination with CIK shows short-term efficacy on hepatocellular carcinoma through inducing specific anti-tumor immunity and can be an effective adjuvant therapy for hepatocellular carcinoma.

Objective To optimize the concentration of a microRNA-21 (miR-21) inhibitor and a miR-494 mimic for the transfection of A375 human melanoma cells,and to estimate the effect of the miR-21 inbihitor and miR-494 mimic on the proliferation of A375 cells.Methods A miR-21 inbihitor and a miR-494 mimic were designed and constructed.To optimize the concentration of the miR-21 inbihitor and miR-494 mimic for transfection,six concentrations (70-250 nmol/L) of the inbihitor and mimic were transfected into A375 cells separately by using LipofectamineTM2000.Then,quantitative fluorescence-based PCR was performed to determine the expression of miR-21 and miR-494 in A375 cells.Some A375 cells were classified into five groups:Mock blank control group remaining untransfected,miR-21 inhibitor group transfected with the miR-21 inhibitor,miR-21 control group transfected with the miR-21 inhibitor negative control,miR-494 mimic group transfected with the miR-494 mimic,and miR-494 control group transfected with the miR-494 mimic negative control.Mter another 48-hour culture,the cells were collected for the analysis of cell apoptosis and cycle by using flow cytometry.Meanwhile,Cy5-labelled miR-494 mimic negative control was transfected into A375 cells for the evaluation of the transfection efficiency by using an inverted fluorescence microscope.Results miRNAs were successfully extracted from A375 cells.As quantitative PCR revealed,the A375 cells transfected with the miR-21 inhibitor at 120 nmol/L showed the lowest expression level (2-△△Ct) of miR-21 (average:0.80; range:0.65-0.92),and those transfected with the miR494 mimic at 250 nmol/L displayed the highest expression level of miR-494 (average:126.82; range:111.52-144.22).The transfection efficiency in A375 cells was higher than 90％.Compared with the corresponding negative control groups,the miR-21 inhibitor group and miR-494 mimic group showed increased apoptosis rate ((27.74 ± 1.39)％ vs.(12.93 ± 0.65)％,(34.30 ± 2.35)％ vs.(15.54 ± 1.02)％,both P ＜ 0.01),percentage of G1-phase cells ((61.61 ± 3.25)％ vs.(50.34 ± 5.62)％,(61.05 ± 3.17)％ vs.(49.95 ± 2.58)％,both P＜ 0.05),but decreased proliferation index ((38.39 ± 3.25)％ vs.(49.66 ± 5.62) ％,(38.95 ± 3.17)％ vs.(50.05 ± 2.58)％,both P ＜ 0.05).Conclusions Both the miR-21 inhibitor and miR-494 mimic can promote the G1-phase arrest and apoptosis in A375 cells,and miR-21 may act as a protooncogene accelerating the proliferation of A375 cells,while miR-494 may founction as a tumor suppressor inhibiting the proliferation of A375 cells.

Objective To elucidate the immunologic mechanism and clinical effect of high intensity focused ultrasound (HIFU) combined with dendritic cell and cytokine induced killer cell (DC-CIK) immunotherapy on patients with pancreatic cancer.Methods Seventy-two pancreatic cancer patients were divided randomly into 2 equal groups,one treated with HIFU only the other treated with HIFU and DC-CIK immunotherapy.Ultrasound imaging and a variety of immunological indexes were recorded before and after treatment and the clinical effects in the two groups were compared.Moreover,autogenous tumor cells were isolated from the combination therapy group and the killing activity of DC-CIK which loaded tumor antigen processed by HIFU on autogenous tumor cells was observed.Results Tumor antigen processed by HIFU can improve the killing activity of DC-CIK on autogenous tumor cells.After treatment,the immunological indexes,of all patients were better than before treatment.(58.26 ± 17.97 versus 52.15 ± 14.22 pg/ml with IL-12 22.14 ± 6.39 versus 17.36 ± 5.73 ng/ml with HSP70 and 0.94 ± O.34 versus 1.32 ± O.61 ng/ml with TGF-β,P ＜ 0.05 ) ; The combination group was significantly better than the HIFU group with regard to the average scores of quality of life (75.89 ± 19.65 versus 67.22 ± 16.34,P＜0.05),pain (3.15 ±0.82 versus 3.59 ± 1.04,P ＜0.05),tumor markers (107.55 ±27.58 versus 123.63 ±34.12 U/ml) and survival time (18.92±6.47 versus 13.36 ±5.78 mos).Conclusion HIFU can improve the immunologic status and anti-tumor response in patients with pancreatic cancer.HIFU combined with DC-CIK has good synergistic therapeutic effect for treating pancreatic cancer.

Objective To investigate the expression of microRNA-21(miRNA-21)in two human malignant melanoma cell lines,A375 and M14,and its effect on the viability of these cells.Methods The expression of human miRNA-21 was assessed by quantitative fluorescent PCR in A375 and M14 cells.Then,three concentrations(90,180,270 nmol/L)of miRNA-21 inhibitor and were transfected both cells and a negative control were transfected a mixture of LipofectamineTM 2000 reagent,respectively.After another 3-day culture,the proliferation of cells was detected by cell counting kit-8,and R value was calculated to denote the relative activity of cells.Statistical analysis was carried out by SPSS13.0.Results The expression of miRNA-21 was higher on A375 cells than that on M14 cells with the average value of 2-deltaCT being 1.2928±0.1509 vs 0.1894±0.1803.With miRNA-21 inhibitor at the concentration of 90,180,270 nmol/L,the activity of A375 cells was significantly lowered in comparison with that in the control group,with the R value being 0.7362±0.1662.0.7248±0.3204 and 0.6767±0.2998 respectively(all P＜0.01).However,in the case of M14 cells,cell activity was only suppressed by miRNA-21 inhibitor at 90 nmol/L with the R value being 0.7295±0.1478.and no significant inhibition was observed with the inhibitor at 180 or 270 nmol/L (both P＞0.05).Conclusions miRNA-21 is expressed on human melanoma cell lines,A375 and M14,at different levels,with a promoting effect on the proliferation of both cells.Moreover,miRNA-21 may act as an oncogene-like gene via down-regulating the expression of some tumor-inhibiting factors.

Objective To explore the immune effects of high intensity focused ultrasound (HIFU) in the treatment of pancreatic carcinoma and to investigate the imaging methods to evaluate HIFU's efficacy.Methods A total of 32 patients with pancreatic carcinoma treated by HIFU were enrolled.The freeze-thaw antigen was prepared by freezing and thawing the cancer cells.HIFU antigen was prepared by cancer cells sonicated by HIFU.The killing effects of no antigen activated dendritic cells (DC) induced T lymphocyte (DC-T),freeze-thaw antigen activated DC induced T lymphocyte (freeze-thaw antigen-DC-T) and HIFU activated DC induced T lymphocyte (HIFUantigen-DC-T) in autologous pancreatic cancer cells were detected by lactic dehydrogenase kit.The changes of immune indexes [heat shock protein 70 (HSP70),T helper lymphocyte Thl/Th2 and transforming growth factor-β (TGF-β)] before and after H IFU treatment were determined by enzymelinked immunosorbnent assay (ELISA) method.The changes of clinical efficacy indexes [visual analogue scale (VAS),performance status (PS) and carbohydrate antigen (CA) 19-9] before and after HIFU treatment were compared.The instant and recent (two months) efficacy of HIFU treatment were evaluated by contrast enhanced ultrasonograph (CEUS) and computed tomography (CT).The line q test was performed for comparision between groups.t-test was applied for comparision before and after treatment.Results Compared with freeze-thaw antigen,the killing effect of HIFU antigen-DC-T in autologous pancreatic cancer cells was higher (40.24％ ± 10.56％ vs 46.93％±13.26％,q=3.44,P＜0.05).HSP70 [(17.31±4.75) ng/mlvs (22.84±5.56) ng/ml],Th1/Th2 (1.24±0.36 vs 1.47±0.31),TGF-β [(1.39±0.41) ng/ml vs (1.04±0.38) ng/ml],VAS (3.97±1.32 vs 3.26±1.18),PS (2.76± 1.02 vs 2.21±0.86) and CA19-9 level[(135.39±37.45) U/ml vs (114.82±30.51) U/ml] improved after HIFU treatment compared with those before treatment (t=4.278,2.739,3.542,2.268,2.332 and 2.409,allP＜0.05).CEUS and CT showed that blood supply and the volume of the tumors reduced after HIFU treatment.Conclusions HIFU is effective in treating pancreatic carcinoma,improving immune status of patients and enhancing antitumor response.CEUS can real-time evaluate the efficacy of HIFU treatment.