Health Policy ; Humans ; Italy ; Occupational Health Services ; organization & administration ; Safety Management

Finland ; Health Policy ; Humans ; Occupational Health Services ; organization & administration ; Safety Management

Objective The aim of this study is to investigate CD+8 natural killer T cell receptors NKG2D and NKG2A expression in peripheral blood of patients with lung cancer and discuss the relation between imbalance expression of NKG2A and NKG2D and tumor immune escape. Methods Flow cytometry was used to determine the percentage of NKG2D and NKG2A-expressing of CD+8 NKT cells in peripheral blood of 95 untreated lung cancer patients and 50 healthy controls. Results NKG2D was lower expressed on CD+8 NKT in lung cancer, the level of NKG2D in patients (77.07±5.77) % was significantly lower than that in the controls (84.13±4.49) % (t =8.14, P <0.05). In the TNM stage, the level of NKG2D in patients of Ⅰ-ⅢA, ⅢB, Ⅳ stage were (81.07±5.02) %, (76.95 ±4.70)%, (72.80±5.16) %, respectively, the level of NKG2D was significantly decreased in order (F =18.74, P <0.05). NKG2A was expressed higher on CD+8 NKT in lung cancer, the level of NKG2A in patients (33.58±8.82) % was significantly higher than that in the controls (25.31 ±8.38) % (t =-5.46, P<0.05). In the TNM stage, the level of NKG2A in patients of Ⅰ - Ⅲ A, ⅢB, Ⅳ stage were (25.10±6.93) %, (33.24±3.76) %, (43.64±6.10) %, respectively, the level of NKG2A was significantly increased in order (F =75.73,P <0.05). Conclusion Imbalance expression of NKG2A and NKG2D may restrain the function of CD+8 NKT cell of lung cancer patients in peripheral blood and that may be one of important factors in tumor immunological escape.

Objective To observe the changes of CD+8CD+28,CD+8CD-28 and CD+4CDhigh25CDlow127 regulate T (Treg)cellsin peripheral blood of lung cancer patients,and to analyze the correlation between CD+8CD-28 and CD+4CDhigh25CDlow127 Treg cells to reveal the role and clinical significance of them in lung cancer patients.Methods Flow cytometry was applied to evaluate the level of CD+8CD+28,CD+8CD-28 and CD+4CDhigh25CDlow127 Treg cells in peripheral blood of 60 untreated lung cancer patients and 60 healthy controls group.The association of each term with clinical features was analyzed.Results The percentage of CD+8CD-28 and CD+4CDhigh25CDlow127 Treg cells in lung cancer group[(58.430:15.749) ％,(7.365±2.025) ％]was significantly higher than those in healthy group [(41.057±15.436)％,(6.648±1.669)％,(t=6.102,P＜0.05;t=2.115,P＜0.05)],while the percentage of CD+8CD+28cells is lower(41.570±15.739)％ than that in healthy group[(58.700±15.298)％,(t=-6.043,P＜0.05)].No close associations were found between three index and gender,age,and biological characteristics.With the increase of TNM stage,The percentage of CD+4CDhigh25 CDlow127 Treg cells increased gradually,which was remarkably higher in patients rith stage Ⅳ than that with stage ⅢA(t=-3.898,P＜0.05).The percentage of CD+4CDhigh25 CDlow127 Tregcells was uncorrelated with CD+8CD-28 cells(r=-0.169,P＞0.05).Conclusion The higher percentage of CD+8CD-28 and CD+4CDhigh25 CDlow127 Treg cells,the lower percentage of CD+8CD+28 cells may be the important reasons of immune suppression in lung cancer patients.Though there is no correlation between CD+8CD-28 and CD+4CDhigh25 CDlow127 Treg cells,it is may be helpful to understand immunologic function and it may look for more specific therapy and provide a new reference in the prognosis of lung cancer.

Objective To correlate between the IQQA(R)-Liver system in planning liver resection and the actual operation performed by surgeons.Methods The data on 65 patients were retrospectively studied.Their preoperative enhanced CT images were analyzed by the IQQA(R)-Liver system to determine the operative plan (Group Q) including the operative technique,the major vessels which required to be transected and the virtual liver resection volume.The above results and the corresponding data collected from the actual operation (Group S) were statistically analyzed to find out whether there was any correlation between them,thus determining the clinical significance of the IQQA(R)-Liver system in preoperative evaluation.Results Group Q and Group S had good correlation in the operative techniques (O) and in the major vessels that required to be transected (A) (uO =0.835,uA =0.893) with no statistical difference between the 2 groups (PO =0.494,PA =0.331).The virtual liver resection volume was 633.96 ± 512.06 (78.30 ～2 559.38)cm3.Conclusion Preoperative evaluation by the IQQA(R)-Liver system had significance in planning partial hepatectomy.

Objective To investigation the expressions of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors in non-small cell lung cancer (NSCLC) and their clinical significances.Methods The serum expression levels of TRAIL in 79 cases of NSCLC and 80 cases of normal subjects were detected by enzyme-linked immunosorbent assay (ELISA).The expressions of TRAIL-R2 and TRAIL-R4 in 42 cases of NSCLC and matched normal tissues were detected by immunohistochemistry.The relationships among TRAIL,TRAIL-R2,TRAIL-R4 and clinicopathologic features of NSCLC were analyzed.Results The expression of TRAIL in NSCLC patients was lower than that in normal human [(994.3 ±293.0)ng/ml vs.(1 141.7 ±266.1)ng/ml,t =3.29,P =0.00].The expression of TRAIL was closely correlated with clinical stage (F =2.28,P =0.00) and differentiated degree (t =5.76,P =0.00).The positive expression rate of TRAIL-R2 in NSCLC was 73.8％ (31/42),significantly lower than that in the normal tissue 100.0％ (42/42) (x2 =3.88,P =0.05).The expression of TRAIL-R2 was closely correlated with clinical stage (x2 =27.89,P=0.00) and differentiated degree (x:=9.50,P =0.00).The positive expression rate of TRAIL-R4 in NSCLC was 81.0％ (34/42),significantly higher than that in the normal tissue 50.0％ (21/42) (x2 =7.34,P =0.01).The expression of TRAIL-R4 was also closely correlated with clinical stage (x2 =17.82,P =0.00) and differentiated degree (x2 =4.47,P =0.03).There was a negative correlation between the expression of TRAIL-R2 and TRAIL-R4 in NSCLC (r =-0.67,P=0.01).Conclusion The decrease of TRAIL and TRAIL-R2 and increase of TRAIL-R4 expression may promote the occurrence and development of NSCLC,and they may provide targets for clinical treatment of NSCLC.

Objective: To observe the role of rmh-TNF in enhancing the anti-angiogenesis effect of cisplatin on Lewis lung carcinoma in the mice.Methods: Lewis lung carcinoma model was established in C57BL/6 mice.Sixty model mice were randomly divided into 4 groups: control group,rmh-TNF group(1500000 U/kg),cisplatin group(6.15 mg/kg), and rmh-TNF plus cisplatin group.Twelve days after implantation of cancer cells,different drugs were injected intra- tumorallv for 3d.The expression of hypoxia inducible factor-1?(HIF-1?)gene in the tumor was identified by RT-PCR. Immunohistochemistry(IHC)image analysis was performed to determine the vascular endothelial growth factor(VEGF) and kinase domain region receptor(KDR)expression and the microvessel density(MVD).Expression of matrix metallo- proteinase-2(MMP-2)was detected by flow cytometry.Results: The MVD values in the control group,the rmh-TNF group,the DDP group and the combination group were(24.76?1.28),(18.95?1.22),(19.53?1.15),(10.43?1.05),respectively,with those of the rmh-TNF and DDP groups significantly lower than that of the control group and higher than that of the combination group(all P

Objective To evaluate the recognition and uptake of transglutaminase 3 (TG3) by dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) receptors on the membrane surface of DC-SIGN-transfected human embryonic kidney (HEK) 293T cells and monocytederived dendritic cells (MDDCs).Methods The eukaryotic expression vector pGCMV-enhanced green fluorescent protein (EGFP) containing DC-SIGN gene fragments was transfected into HEK293T cells to prepare DC-SIGN-EGFP-HEK293T cells by using liposome transfection method.CD14+ monocytes were isolated from peripheral blood samples by magnetic bead-based negative selection,and then were induced by granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) to prepare MDDCs.Laser confocal microscopy and flow cytometry were performed to evaluate the recognition and uptake of TG3 protein by DC-SIGN receptors on the surface of HEK293T cells and MDDCs.MDDCs treated without Alexa Fluor 647 dye-tagged TG3 served as blank control group,and those treated with Alexa Fluor 647 dye alone served as negative control group.Results After co-culture with TG3 for 3 hours,laser confocal microscopy and flow cytometry both showed that TG3 could be recognized by and uptaken through DC-SIGN receptors into HEK293T cells and MDDCs.Flow cytometry also revealed that the binding of TG3 to MDDCs could be partially blocked by DC-SIGN blocking antibodies.Neither the negative control group nor the blank control group showed the recognition and binding of TG3 to HEK293T cells and MDDCs.Conclusion TG3 can serve as a kind of autoantigen to be recognized and bound by DC-SIGN receptors,followed by uptake by dendritic cells.

Objective To investigate the expression and clinical significance of Th17 and Tc17 cells in peripheral blood of lung cancer patients.Methods The percentages of Th17 cells and Tc17 cells in 60 lung cancer patients and 40 healthy controls were evaluated by flow cytometry analysis (FCM).Results The percentages of Th17 cells [(1.795±0.623) ％] and Te17 cells [(0.865±0.357) ％] in lung cancer group were significandy higher than those in controls [(1.405±0.256) ％,(0.640 ±0.204) ％],(t =28.944,P ＜ 0.001,t =14.051,P ＜ 0.001).Furthermore,there was a positive correlation between Th17 cells and Tc17 cells in the two groups (lung cancer group r =0.770,P ＜ 0.05,control group r =0.532,P ＜ 0.05).The percentages of Th17 cells and Tc17 cells were closely associated with clinical stage (F =4.882,P =0.011,F =3.633,P =0.033),but not connected with pathological types (P ＞ 0.05,P ＞ 0.05).Conclusion The overexpression of Th17 and Tc17 may be involved in the occurrence and development of lung cancer,which can be used as new indicators for immunologic function of lung cancer patients,and provide a reference in monitoring the disease.

OBJECTIVETo investigate the effect of Gli1 gene silencing by RNA interference (RNAi) on proliferation of K562 cells and its mechanisms.

METHODSThe small interference RNA (siRNA) was synthesized in vitro. K562 cells were transfected with Gli1 siRNA by the way of lipofection (lipofectamine 2000). Non-specific siRNA transfected cells were used as control. Transfection efficiencies of different siRNA concentrations were detected by flow cytometry and the best siRNA concentration was selected. The silencing effect of siRNA was demonstrated by real time PCR and Westem blot analysis. Cell proliferation was measured by MTT method, cell cycle by PI assay, c-myc and p21 mRNA level was detected by real time PCR analysis.

RESULTSTransfection efficiency of siRNA was increased in a dose-dependent manner when siRNA concentration was below 200 pmol, and the highest transfection efficiency reached (80.11 ± 5.63)%. Both the mRNA and protein level of Gli1 was down-regulated in Gli1 specific siRNA group, the mRNA level was (52.60 ± 3.57)% of that of control group after 24 h (t = 20.33, P < 0.01) and the protein level was (79.31 ± 5.58)% of that of control group after 48 h (t = 6.54, P < 0.01). The cell proliferation rate in Gli1 siRNA group was (94.41 ± 3.58)% (t = 2.40, P = 0.05) and (90.22 ± 3.34)% (t = 4.37, P < 0.01) of that of control group after 24 h and 48 h, respectively. G(2)/M cell cycle arrest was observed, the mRNA level of c-myc was down-regulated while p21 was up-regulated in Gli1 siRNA group after 24 h and 48 h (P < 0.05).

CONCLUSIONSTargeted silencing of Gli1 gene by RNAi inhibits the proliferation of K562 cells, which acts through the down-regulation of c-myc and up-regulation of p21 expression.

Cell Proliferation ; Gene Silencing ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transcription Factors ; genetics ; Transfection ; Zinc Finger Protein GLI1