Value of needle confocal laser microendoscopy combined with endobronchial ultrasound bronchoscopy in the diagnosis of hilar and mediastinal lymph node lesions.

Endobronchial ultrasound bronchoscopy (EBUS) and needle confocal laser endomicroscopy (nCLE) are techniques for screening benign and malignant lesions of the hilar and mediastinal lymph node (HMLN). This study investigated the diagnostic potential of EBUS, nCLE, and combined EBUS and nCLE in HMLN lesions. We recruited 107 patients with HMLN lesions who were examined by EBUS and nCLE. A pathological examination was performed, and the diagnostic potential of EBUS, nCLE, and combined EBUS-nCLE approach was analyzed according to the results. Among the 107 cases of HMLN lesions, 43 cases were benign and 64 cases were malignant on pathological examination, 41 cases were benign and 66 cases were malignant on EBUS examination; 42 cases were benign and 65 cases were malignant on nCLE examination; 43 cases were benign and 64 cases were malignant on combined EBUS-nCLE examination. The combination approach had 93.8% sensitivity, 90.7% specificity, and 0.922 area under the curve, which was higher than those of EBUS (84.4%, 72.1%, and 0.782, respectively) and nCLE diagnosis (90.6%, 83.7%, and 0.872, respectively). The combination approach had a higher positive predictive value (0.908), negative predictive value (0.881), and positive likelihood ratio (10.09) than that of EBUS (0.813, 0.721, and 3.03, respectively) and nCLE (0.892, 0.857, and 5.56, respectively), whereas, the negative likelihood ratio was lower than that for EBUS (0.22) and nCLE (0.11). No serious complications occurred in patients with HMLN lesions. To summarize, the diagnostic efficacy of nCLE was better than EBUS. The EBUS-nCLE combination is a suitable approach for diagnosing HMLN lesions.

[Role of MG132, a proteasome inhibitor, in inducing expression of costimulatory molecules CD86 in leukemic cells and its effect on allogeneic mixed lymphocyte reaction].

OBJECTIVE: To investigate the role of proteasome inhibitors MG132 in the inducing the expression of the costimulatory molecules CD80 and CD86 in leukemia cells and its effect on allogeneic mixed lymphocyte reaction. METHODS: Acute myelocytic leukemia cells of the line HL-60 and chronic myelocytic leukemia cells of the line K562 were cultured. 7-AAD staining and flow cytometry (FC) were used to examine the viability of the cells. MG132, a proteasome inhibitor, of the concentrations of 2 or 3 micromol/L was added into the culture fluid of HL-60 cells for 24 h and 48 h respectively and then annexin V/7-AAD staining and FC were used to detect the apoptosis of the cells. HL-60 and K562 cells treated with 1 micromol/L MG132 for 24 h and 48 h respectively, anti-CD80 and anti-CD86 antibodies were added, then FC was used to detect the expression of CD80 and CD86. The mRNA expression of CD86 in the HL-60 cells treated with 1 micromol/L MG132 was examined by RT-PCR. HL-60 and K562 cells were treated by 1 micromol/L MG132 for 48 h and then underwent irradiation of 75 Gy Co-60 to kill the cells with their antigenicity preserved. Peripheral blood mononuclear cells (PBMNC) of healthy volunteers, as reactive cells, were isolated and inoculated into the Co-60 treated HL-60 and K532 cells of different concentrations, as stimulating cells, for 5 d, CCK-8, a new agent to detect the cell viability, was added for 4 h, and then the A value of absorbance was measured at the wave length of 450 nm of enzyme labeling instrument. Control groups were set up for all tests. RESULTS: The cell viability rates of the HL-60 cell treated with 1 micromol/L MG132 for 24 h and 48 h were 92.95% and 85.87% respectively. The apoptotic rats of the HL-60 cells treated with MG132 were increased dose- and time-dependently. Before MG132 treatment K562 cells did not express CD86, and the CD86 expression of the HL-60 cells was up-regulated time-dependently (all P < 0.01). The mRNA expression of CD86 in the HL-60 treated with MG132 was up-regulated time-dependently (P < 0.01). CKK8 test showed that the proliferation level of PBMNC gradually increased along with the concentration of HL-60 cells treated with MG132 and reached its peak when the concentration of the HL-60 cells was 1 x 10(5) (P < 0.01). No remarkable proliferation of PBMNC was seen in the K562 groups no matter if the HL-60 cells had been treated with MG132. CONCLUSION: MG132 induces the expression of costimulatory molecule CD86 in the HL-60 cells, thus improving the proliferation of PBMNC.

Effects of targeting magnetic drug nanoparticles on human cholangiocarcinoma xenografts in nude mice.

BACKGROUND: Targeting is a new therapeutic tool for malignant tumor as a result of combining nanotechnology with chemotherapeutics. The aim of our study was to investigate the effects of magnetic nanoparticles enveloping a chemotherapeutic drug on human cholangiocarcinoma xenografts in nude mice. METHODS: The human cholangiocarcinoma xenograft model was established in nude mice with the QBC939 cell line. The nude mice were randomly assigned to 7 groups. 0.9% saline or magnetic nanoparticles, including high (group 2), medium (group 4) and low (group 5) dosages, were given to nude mice through the tail vein 20 days after the QBC939 cell line was implanted. Calculations were made 35 days after treatment in order to compare the volumes, inhibition ratios and growth curves of the tumors in each group. Mice in each group were sacrificed randomly to collect tumor tissues and other organs for electron microscopy and pathological examination. RESULTS: The high and medium dosage groups were significantly different from the control group (P<0.05). The tumor inhibition ratios for the high, medium and low dosage groups were 39.6%, 14.6% and 7.9%, respectively. The tumor growth curve of groups 5, 4, and 2 changed slowly in turn. The high and medium groups showed cell apoptosis under an electron microscope. CONCLUSION: Magnetic nanoparticles can inhibit the growth of human cholangiocarcinoma xenografts in nude mice.

[Effects of triptolide and TNF-alpha on the expression of VEGF in Raji cells and on angiogenesis in ECV304 cells].

In order to study the relation of antitumour mechanisms of triptolide with neovascularization, the effect of triptolide and tumour necrosis factor (TNF)-alpha on the expression of vascular endothelial growth factor (VEGF) in Raji cell lines and their effect on angiogenesis in human umbilical vein endothelial cells (HUVECs)-derived cell line ECV304 were investigated. The inhibitory rate of cells treated by triptolide detected by MTT; the ELISA was employed to study the VEGF content secreted by Raji cell lines; angiogenesis was tested by network formation of endothelial cells on Matrigel, and the mRNA level of VEGF was measured by RT-PCR. The results showed that treatment of Raji cells with triptolide resulted in significantly enhanced antiproliferative effects in dose- and time-dependent manner. The content of VEGF secreted by Raji cells was increased by TNF-alpha and was suppressed by triptolide (P < 0.01). The mRNA expressions of VEGF(165) and VEGF(121) (containing 165 and 121 amino acid residues, respectively) could be detected in all fractions. TNF-alpha augmented the expression of VEGF(165) and VEGF(121) mRNA when triptolide reduced the expression (P < 0.01). No network and cord were formed in control and triptolide group. There was tube formation on matrigel in the supernatants of Raji culture group and the supernatants groups treated by VEGF and TNF-alpha in Raji cell. It is concluded that the expressions of VEGF in Raji cells are increased by TNF-alpha and suppressed by triptolide. VEGF and TNF-alpha induce angiogenesis and triptolide inhibits angiogenesis in ECV304 cells.