Objective To evaluate the acute and delayed killing effect of high intensity focused ultrasound (HIFU) on Echinococcus granulosus(E. granulosus)protoscolices in vitro.Methods E. granulosus protoscolices were treated with different dosage of effective power(0,25,50,100,200,250 W)and time(5,10,20,30,40,50,60 s)of HIFU in vitro to obtain the dosage-effect curves.Then the survival pmtoscolices were incubated,and the mortality of each group was counted daily.The protoscolicidal effects were investigated by trypan blue exclusion assay.Results Compared with the untreated group,the Vitality of E.granulosus protoscolices significantly decreased immediately after treated by HIFU of different dosage(F=5201.59 vs 1865.65,P＜0.05),there were the interaction both different dosage and time(F=214.50,P＜0.05).The protoscolices were broken into pieces by HIFU of 250 W×40 s,whereas the growth of the surviving protoscolices after exposed to HIFU was obvious suppressed.Both the acute killing effect and the delayed inhibitory effect showed a dosage-dependant manner.The inhibitory effect increased along with the increased dosage of HIFU(P＜0.05).The inhibitory effect in 50 W×10 s group was stronger than 25 W×20 s group(P＜0.05).The mortality was increased in parallel with the increase of HIFU dosage.Conclusions HIFU show an effective immediately killing effect,as well as a growth-inhibiting effect on the E.granulosus protoscolices in vitro.

OBJECTIVETo investigate the ectopic osteogenesis potential of human natural bone derived material combined with human bone marrow mesenchymal stem cells (MSCs).

METHODSCell-scaffold complexes were implanted subcutaneously into the left back of the nude mice, and human natural bone derived material were implanted into the right back as control group. The mice were killed respectively on the postoperative 2 weeks, 4 weeks and 8 weeks. The macroscopic, histopathological, alkaline phosphatase (ALP) activity assay methods were performed to assess the ectopic osteogenesis potential.

RESULTSThe cartilaginous osteogenesis were observed in both deproteinated bone and decalcified bone, and the more new bone tissue formed gradually as the time went by after implantation. ALP activity become stronger followed with the time (P < 0.05), and compared with the decalcified bone, deproteinated bone displayed stronger ALP activity (P < 0.05).

CONCLUSIONThe MSCs and human natural bone derived material can be used as good seed cells and scaffold materials respectively to construct tissue-engineered bone, and as the scaffold material, deproteinated bone has better osteogenesis ability than decalcifed bone.

Animals ; Bone and Bones ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; Mice ; Mice, Nude ; Osteogenesis ; Tissue Engineering

OBJECTIVETo explore the feasibility of multi-dimensional and comprehensive evaluation on the quality of life among rural elderly population in Anhui province.

METHODS5652 rural elderly people aged above 65 in Anhui province were selected by cluster sampling method and were studied by cross-sectional study through a questionnaire on health information. The quality of life was evaluated by comprehensive evaluation method.

RESULTSThe total score of satisfactory quality of life in the studied rural elderly people was 0.1432 +/- 0.5170, while not satisfied was -0.2521 +/- 0.6081, with significant difference between the two groups (F = 666.221, P < 0.0001). There was positive correlation between subjective satisfaction and total score of quality of life, with r(s) = 0.345 (P < 0.0001). The results of logistic regression analysis between comprehensive index of quality of life and subjective satisfaction indicated that filial piety, income, sleeping condition, chronic disease, nutrition status, economic dominance in the family, amusement activities etc. were important factors influencing the quality of life.

CONCLUSIONIt was feasible to evaluation on the quality of life by comprehensive evaluation method.

Activities of Daily Living ; Aged ; Aged, 80 and over ; China ; Evaluation Studies as Topic ; Female ; Health Status ; Humans ; Male ; Personal Satisfaction ; Quality of Life ; Regression Analysis ; Rural Health ; standards ; statistics & numerical data ; Socioeconomic Factors ; Surveys and Questionnaires

OBJECTIVETo evaluate the role of human leukocyte antigen G (HLA-G) in the better effect of allogenetic bone marrow transplantation than that of peripheral blood stem cell transplantation.

METHODSFlow cytometry was used to detect the expression of membrane-bound HLA-G (mHLA-G) on donor peripheral blood (PBC) or bone marrow (BM) mononuclear cells. The levels of soluble HLA-G (sHLA-G) in the plasma and bone marrow fluid were determined using enzyme-linked immunosorbent assay (ELISA) before and after granulocyte colony-stimulating factor (G-CSF) mobilization.

RESULTSThe mean levels of mHLA-G after G-CSF mobilization in the PBC and BM were significantly higher than that before G-CSF mobilization (P=0.001 and 0.000), but the plasma levels of sHLA-G showed no significant changes after the mobilization (P=0.279). The mean levels of sHLA-G in the BM fluid significantly increased (P=0.002) to a level higher than that in the PBC after G-CSF mobilization (P=0.004).

CONCLUSIONHLA-G plays an important role in immune tolerance after hematopoietic stem cell transplantation with G-CSF mobilization.

Adult ; Bone Marrow Transplantation ; immunology ; Female ; Granulocyte Colony-Stimulating Factor ; pharmacology ; HLA Antigens ; immunology ; metabolism ; HLA-G Antigens ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; immunology ; metabolism ; Humans ; Immune Tolerance ; Male ; Middle Aged

OBJECTIVETo evaluate the impact of luteinizing hormone-releasing hormone (LHRH) on the protection of thymic function after allogenic hematopoietic stem cell transplantation (allo-HSCT).

METHODSMurine model of MHC mismatched allogeneic HSCT (C57BL/6-->BALB/c) was established. The severity of acute graft-versus-host-disease (GVHD) was assessed according to a clinical scoring system. The intra-cellular levels of IFN gamma, TNFalpha and IL-1 beta in thymocyte were analyzed by protein array and thymic function by quantification of signal-joint TCR rearrangement excision circles (sjTRECs).

RESULTSAll recipients in group A (allogeneic mice), B (allogeneic LHRH castrated-mice) and C (syngenic mice) achieved hematopoietic reconstitution. White blood cell (WBC) over 1.0 x 10(9)/L in groups A, B and C were on day (11.2 +/- 1.4), day (9.8 +/- 0.6) and day (9.7 +/- 0.7), respectively (P = 0.003, 0.002). The onset of acute GVHD in group B was (14.1 +/- 0.7) d and in group A was (11.4 +/- 1.2) d (P = 0.000). All mice in groups A and B developed acute GVHD. No mice occurred aGVHD in group C. The average scores of acute GVHD in groups A and B were (9.1 +/- 0.7) and (5.1 +/- 1.0), respectively (P = 0.000). The levels of IFN gamma, TNFalpha and IL-1 beta in control group were (2.3 +/- 2.5) ng/ml, (1.7 +/- 1.1) pg/ml and (1.8 +/- 1.2) pg/ml, respectively. The IFN gamma levels in groups A, B and C were (10.5 +/- 2.1) ng/ml, (6.7 +/- 2.1) ng/ml and (5.2 +/- 3.3) ng/ml, TNFalpha levels were (7.0 +/- 2.6) pg/ml, (4.3 +/- 0.8) pg/ml and (3.0 +/- 1.8) pg/ml, and IL-1 beta levels were (24.9 +/- 9.0) pg/ml, (17.4 +/- 3.9) pg/ml and (10.8 +/- 3.1) pg/ml, respectively. There were significant differences in the levels of cytokines between group A and the control group (P = 0.000, 0.000, 0.000). The levels of cytokines in group B were significantly higher than those in control group (P = 0.000, 0.003, 0.000). The levels of IFN gamma and IL-beta in group C were significantly higher than those of in control group (P = 0.015, 0.013), and so did in group A than in group B (P = 0.002, 0.002, 0.004), and in group A than in group C (P = 0.000, 0.000, 0.000). The analysis of linear regression showed that the average levels of IFN gamma and TNFalpha paralleled with aGVHD scores (r(2) = 0.359, P = 0.045; r(2) = 0.228, P = 0.019). The average sjTRECs copies/1000 PBMNCs were (39.4 +/- 44.7) in the control group and (12.3 +/- 13.0), (58.0 +/- 71.8) and (19.6 +/- 14.6) in groups A, B and C, respectively. There was no significant difference in the multiple comparisons of peripheral blood levels of sjTRECs among these four groups (P = 0.468).

CONCLUSIONIFN gamma, TNFalpha and IL-1 beta might be involved in the damage to the thymus by acute GVHD. Sex steroid inhibitor can not only reduce the severity of thymic damage after allo-HSCT, but also reduce the severity of aGVHD and the mechanism might be associated with the reduction of intra-cellular levels of IFN gamma in thymocyte.

Animals ; Castration ; methods ; Female ; Gonadotropin-Releasing Hormone ; therapeutic use ; Graft vs Host Disease ; pathology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Interferon-gamma ; metabolism ; Interleukin-1beta ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Thymus Gland ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To investigate the effects of DL-3-n-Butylphthalide(NBP)on proliferation and apoptosis of 1-methyl-4-phenyl-pyridinium (MPP +)-induced SH-SY5Y cells, and mechanisms via mixed lineage kinase 3 (MLK3) signaling pathway. Methods The SH-SY5Y cells were divided into control group,MPP+group,NBP group and URMC-099 group,that cultured normally,with 1 mmol/L MPP+for 24 hours,with 10μmol/L NBP for 3 hours and then with MPP+for 24 hours,and with 200 nmol/L MLK3 inhibitor URMC-099 for 3 hours and then with MPP+for 24 hours,respectively.The morphology of SH-SY5Y cells was observed under inverted phase contrast mi-croscope and the survival rate was measured with 3-(4,5-Cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays.The apoptosis was quantified under flow cytometry with Annexin V/PI fluorescence staining,and the nuclear morphology was observed with Hoechst 33342 staining.The expression of phosphorylated protein of MLK3(p-MLK3),c-Jun N-terminal kinase(p-JNK),extra cellular regulated protein ki-nases(p-ERK1/2)were detected with Western blotting.Results Compared with the control group,the survival rate reduced and apoptosis in-creased in MPP+group(P<0.05),with the increase of p-MLK3 and p-JNK and decrease of p-ERK1/2 d(P<0.05).Compared with MPP+group,the survival rate increased and apoptosis reduced in both NBP and URMC-099 groups(P<0.05),with the decrease of p-MLK3 and p-JNK and increase of p-ERK1/2(P<0.05).Conclusion NBP can decrease the apoptosis and promote the proliferation of SH-SY5Y cells in-duced by MPP+,which may be associated with inhibiting MLK3 signaling pathway,and regulating the downstream p-JNK and p-ERK1/2.

OBJECTIVETo detect the expression of membrane-bound HLA-G (mHLA-G) and serum HLA-G (sHLA-G) in acute leukemia patients and investigate the correlation between HLA-G expression and the occurrence and development of acute leukemia.

METHODSEnzyme-linked immunosorbent assay and flow cytometry were used to detect the expression levels of sHLA-G and mHLA-G in 40 newly diagnosed leukemia cases, 10 refractory and relapsed leukemia cases, and 30 leukemia cases receiving chemotherapy. Ten normal individuals served as the normal control group.

RESULTSThe mean serum level of sHLA-G in normal individuals was 5.87±2.07 ng/ml, as compared to 10.05±6.58 ng/ml in newly diagnosed leukemia patients and 12.32±5.85 ng/ml in refractory and relapsed cases. The mean level of mHLA-G in normal individuals, newly diagnosed cases, and refractory and relapsed cases were (0.29±0.20)%, (0.60±0.44)%, and (0.77±0.41)%, respectively. The mean levels of sHLA-G and mHLA-G were significantly higher in the newly diagnosed cases than that in the normal controls (P<0.05), and significantly higher in patients before chemotherapy than in those with complete remission after chemotherapy (P<0.05).

CONCLUSIONHLA-G expression levels might influence the treatment outcomes and can serve as a prognostic factor for acute leukemia.

Acute Disease ; Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; HLA-G Antigens ; blood ; metabolism ; Humans ; Leukemia ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Young Adult

OBJECTIVETo investigate the effect of granulocyte colony stimulating factor (G-CSF) on myeloid-derived suppressor cells (MDSCs) in the bone marrow and peripheral blood, and explore the relationship between MDSC and graft-versus-host disease (GVHD).

METHODSBone marrow, peripheral blood and peripheral blood stem cells were obtained from 12 healthy hemopoietic stem cell donors before and on day 5 after G-CSF mobilization. Flow cytometry was employed to examine the number of MDSC, and the relationship between MDSC number and the incidence of GVHD was analyzed.

RESULTSIn normal physiological conditions, MDSC could be detected in the peripheral blood and bone marrow with a cell percentages of (1.35±0.35)% and (2.44±1.11)%, respectively, showing a significantly higher cell percentage in the bone marrow (P=0.015). On the 5th day after G-CSF mobilization, the percentage of MDSCs increased to (4.01±1.82)% in the peripheral blood and to (4.38±2.19)% in the bone marrow, showing no significant difference between them (P=0.083). The mobilization caused a significant increase in the number of MDSCs in the peripheral blood (P=0.047) but not in the bone marrow (P=0.761). The number of MDSCs in the collected samples showed a significant inverse correlation to the incidence of GVHD (P=0.048).

CONCLUSIONSMDSCs are present in the peripheral blood and bone marrow of healthy donors, with a greater number in the bone marrow. G-CSF can mobilize the MDSCs from the bone marrow to the peripheral blood to increase number of MDSCs in the peripheral blood, which may contribute to a lowered incidence of GVHD in hematopoietic stem cell transplantation (HSCT).

Adolescent ; Adult ; Bone Marrow Cells ; cytology ; Female ; Graft vs Host Disease ; prevention & control ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; T-Lymphocytes ; cytology ; drug effects ; metabolism ; Young Adult

OBJECTIVETo investigate the expression of T cell receptor (TCR) Vgamma genes in patients with graft-versus-host disease (GVHD) after allogenic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe expression levels of the TCR Vgamma I-III genes in peripheral blood mononuclear cells (PBMNCs) from 18 patients with GVHD following allo-HSCT were determined using real-time fluorescence quantitative PCR, with 12 healthy individuals serving as the normal controls.

RESULTSThe expression level of TCR Vgamma II gene in the PBMNCs from patients with GVHD was significant lower than that in the normal controls. The expression patterns of TCR Vgamma I-III subfamilies also underwent alterations in patients with GVHD, and the expression level of TCR Vgamma II gene was significantly lower than that of TCR Vgamma I gene or TCR Vgamma III gene.

CONCLUSIONThe low expression of TCR Vgamma II subfamily might be related to the pathogenesis of GVHD in patients receiving allo-HSCT.

Adult ; Case-Control Studies ; Female ; Graft vs Host Disease ; genetics ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Young Adult

OBJECTIVETo explore the effect of ricin temperature response gel on breast cancer and its regulatory effect on immune function in rats.

METHODSRicin was purified by chromatography and identified by immunoblotting. The rat subcutaneously transplanted breast cancer model was established. Forty model rats with a tumor diameter of about 3.0 cm were subjected to the study. They were randomized into four groups equally: the model group and three treated groups (blank gel, ricin, ricin-gel) were administered with blank gel, ricin, and ricin temperature response gel via percutaneous intratumor injection, respectively. The tumor was isolated 10 days later for the estimation of tumor inhibition rate (TIR) by weighing, pathologic examination, and detection of tumor apoptosis-associated genes bcl-2 and bax with semiquantitative RT-PCR. Also, peripheral blood was obtained to test T-lymphocyte subsets, the killing function of lymphocytes, and the contents of tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2). The outcomes were compared between groups.

RESULTSThe TIR in the ricin-gel group was 61.8%, with the pathologic examination showing extensive tumor tissue necrosis. Compared with the model group, after ricin temperature response gel treatment, bcl-2 expression was down-regulated, bax expression was up-regulated, CD4+ lymphocytes and CD4+/CD8+ ratio in peripheral blood were increased, the killing function of lymphocytes was enhanced, and the contents of TNF-α and IL-2 were elevated (P < 0.05 or P < 0.01).

CONCLUSIONIntratumor injection of ricin temperature-responsive gel showed significant antitumor effect on breast cancer and could enhance the immune function in the tumor-bearing rat.

Animals ; Antineoplastic Agents ; administration & dosage ; Apoptosis ; drug effects ; CD4-CD8 Ratio ; Disease Models, Animal ; Female ; Gels ; therapeutic use ; Immunohistochemistry ; Immunomodulation ; drug effects ; Injections, Intralesional ; Interleukin-2 ; immunology ; metabolism ; Mammary Neoplasms, Experimental ; drug therapy ; immunology ; pathology ; Random Allocation ; Rats ; Rats, Wistar ; Ricin ; administration & dosage ; Sensitivity and Specificity ; Temperature ; Tumor Necrosis Factor-alpha ; immunology ; metabolism