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Objective To investigate the role of chemokine ligand 19 (CCL19) and protein kinase-B (AKT) signaling pathway in lung cancer development. Methods The human lung adenocarcinoma cell line, A549 cells, in logarithmic growth phase were randomly divided into five groups： blank control group, solvent control group, CCL19 treatment group, AKT inhibition group, and antibody neutralization group. The blank control group received no treatment. The other four groups were treated with dimethyl sulfoxide, CCL19, MK-2206 (AKT inhibitor), and a combination of CCL19 and MK-2206, respectively. Cell viability was assessed using the CCK-8 assay, while cell migration and invasion capabilities were evaluated using the cell scratch and transwell assays. The relative expression levels of Pan-AKT, p-AKT (Ser473), p-AKT (Thr308), E-cadherin (E-cad), N-cadherin (N-cad), and Snail proteins in A549 cells were detected using Western blotting. Lung cancer tissue samples from 60 patients with non-small cell lung cancer (NSCLC) were collected, and the expression of CCL19 and matrix metalloproteinase 9 (MMP9) proteins in the specimens was examined using immunohistochemistry. Results The survival rate of A549 cells in the AKT inhibition group and antibody neutralization group was lower than that in blank control group, solvent control group, and CCL19 treatment group (all P<0.05). The cell scratch assay result showed that the cell migration rate of the CCL19 treatment group was higher at 36.0 and 48.0 hours than those of the blank control group, solvent control group, AKT inhibition group, and neutralizing antibody group (all P<0.05). The Transwell assay result showed that the invasion amount of A549 cells in the AKT inhibition group was less than that in the CCL19 treatment group (P<0.05). Compared with the blank control group, the relative expression of E-cad protein in the CCL19 treatment group decreased, while the relative expression of p-AKT (Ser473), p-AKT (Thr308), N-cad and Snail proteins increased (all P<0.05). The relative expression of p-AKT (Ser473), p-AKT (Thr308), N-cad, and Snail proteins in A549 cells decreased (all P<0.05), and relative expression of E-cad protein increased (all P<0.05) in the AKT inhibition group and antibody neutralization group compared with the blank control group, solvent control group, and CCL19 treatment group. There was no significant difference in the expression of CCL19 and MMP9 in lung cancer tissues of NSCLC patients in Xuanwei City, Gejiu City, and other regions (all P>0.05). The expression of CCL19 and MMP9 in NSCLC patients with lymph node metastasis was higher than in patients without lymph node metastasis (all P<0.01). Conclusion CCL19 can promote the invasion and metastasis of lung cancer cells and induce epithelial-mesenchymal transition. Its expression level is related to lymph node metastasis in NSCLC patients. The AKT signaling pathway may be an important mechanism underlying lung cancer development.
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Objective To determine the optimal cut-off value of critical-care pain observation tool (CPOT) in assessing degree of pain in patients undergone craniotomy, and to determine the sensitivity and specificity of CPOT with this cut-off value. Methods A prospective observational study was conducted in Beijing Tiantan Hospital. A total of 118 patients admitted to intensive care unit (ICU) after craniotomy was consecutively enrolled during August 2014 to August 2015. CPOT and visual analogue scale (VAS) were used to assess the pain before, during and 20 minutes after the removal of central venous catheters, and the difference was compared between two scores at three time points. Receiver operating characteristic (ROC) curve was used to determine the optimal cut-off values for evaluation of the sensitivity and specificity of CPOT. Patients' complaint of pain was considered the gold-standard. Results CPOT values (inter-quartile range) before, during and after the procedure were 0 (0-3), 0 (0-6) and 0 (0-2), respectively; while VAS values were 4 (1, 6), 3 (1, 6) and 4 (1, 6), respectively. CPOT value during the procedure was significantly higher than CPOT values before and after the procedure (both P < 0.01). When the optimal cut-off value of CPOT was 1, CPOT showed the highest Youden index before, during and after the procedure (1.183, 1.515, and 1.438, respectively), and showed high specificity (all 100%) and low sensitivity (18.3% and 43.8%, respectively) when assessing the pain before and after the removal of the catheter. The sensitivity and the specificity were high when assessing the pain during the procedure, the sensitivity was 69.4%, and the specificity was 82.1%. When the optimal cut-off value of VAS was 2 before and during the procedure, and was 4 after the procedure, VAS showed the highest Youden index, 1.568, 1.452, and 1.509, respectively. VAS demonstrated high sensitivity and specificity before, during and after the procedure (sensitivity was 97.2%, 95.2% and 75.0%, respectively; specificity was 59.6%, 50.0% and 75.9%, respectively). The area under ROC curve (AUC) of CPOT before, during and after the procedure were 0.592 [95% confidence interval (95%CI) = 0.490-0.693], 0.778 (95%CI= 0.693-0.863) and 0.719 (95%CI = 0.627-0.811), respectively; the AUC of VAS before, during and after the procedure were 0.846 (95%CI = 0.771-0.920), 0.767 (95%CI = 0.681-0.854) and 0.838 (95%CI = 0.767-0.909), respectively. The AUC of VAS before and after the procedure was significantly higher than the AUC of CPOT (P < 0.001 and P = 0.006), while there was no significant difference between the AUC of VAS and CPOT during the procedure (P = 0.826). Conclusion CPOT can be used to assess the pain during painful procedure, and it shows high accuracy, but with poor evaluation effect on pain in rest.
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Purpose To investigate the sensitivity of BIOMED-2 primer system in T lymphoblastic lymphoma ( T-LBL) and acute lym-phoblastic leukemia ( ALL) patients immunoglobulin ( Ig) and T-cell receptor ( TCR) gene rearrangement, and to analyze the co-rear-rangement pattern. Methods Amplification of rearranged Ig and TCR gene was performed in standard PCR in 35 T-LBL/ALL pa-tients. PCR products were analyzed by heteroduplex and polyacrylamide gel electrophoresis. Results 16 cases (45. 7%) of 35 sam-ples were detected to have TCR gene rearrangements, including 6 cases (37. 5%) of TCRβgene monoclonal rearrangements, 4 cases (25. 0%) of TCRγ gene monoclonal rearrangements, 3 cases (18. 8%) of TCRβ and TCRγ gene double rearrangements, 2 cases (12. 5%) of TCRδ gene monoclonal rearrangements and 1 case (6. 3%) of TCRγand TCRδgene double rearrangements were detec-ted. 4 cases (11. 4%) of 35 samples detected to have clonal immunoglobulin and TCR gene rearrangements. 11 cases (39. 3%) of 28 T-LBL patients were detected to have TCR gene rearrangements, 6 cases (85. 7%) of 7 T-ALL have TCR gene rearrangements. Con-clusions BIOMED-2 multiplex PCR analysis strategy is a useful technique in the T-LBL patients.
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Purpose To discuss the TCR gene rearrangements in the diagnosis of T-cell lymphomas. Methods Formalin-fixed and paraffin-embedded samples including 30 cases of T-cell lymphomas and 30 cases of reactive lymphoid hyperplasia were chosen for ex-tracting genomic DNA and PCR amplification using 56 BIOMED-2 primers. PCR products were analyzed by heteroduplex and polyacryl-amide gel electrophoresis. Results In all 30 cases of T-cell lymphomas, 25 cases (83. 3%) showed TCRβ gene monoclonal rear-rangements, 28 cases (93. 3%) of TCRγ gene monoclonal rearrangements, 4 cases (13. 3%) of TCRδ gene monoclonal rearrange-ments. 29 cases (96. 7%) with TCRβ+TCRγ+TCRδ gene monoclonal rearrangements were detected. but no clonal TCR gene rear-rangements were found in 30 cases of reactive lymphoid hyperplasia. Conclusions The detection of TCR gene rearrangements using BIOMED-2 primers is a useful assistant method for the diagnosis of T-cell lymphomas.
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Objective To investigate the human resources of neurosurgery intensive care unit(NICU)using modified nursing activity score(NAS).Methods NAS was used to investigate the nursing work load of all shifts and compute the needed personnel for every shift,followed by adaption of nursing staff based on the results.Results Based on the scores by NAS,day shift was the heaviest in work load,followed by early night shift and then by late night shift.The differences between day shit and early night shift or late night one were both statistically different(P<0.05 for both).Conclusions The adjustment based on the results by NAS will reduce nursing risk and improve quality of nursing.
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Purpose To study the relationship between Epstein-Barr virus ( EBV) and breast cancer. Methods 246 cases of breast lesions at different development stages were selected and EBV DNA, RNA and protein was used by polymerase chain reaction ( PCR) , in situ hybridization ( ISH) , laser capture microdissection ( LCM) , immunohistochemistry ( IHC) EnVision technology. Results No expression of EBV latent membrane protein LMP1 was detected in all 246 cases of benign and malignant breast lesions. In 12 cases of breast cancer of EBV DNA, carcinoma in situ and breast lesions not EBV DNA was detected by PCR. However, using digoxigenin la-beled EBV DNA probe for the 48 cases ( including 12 cases of breast cancer specimens of positive PCR amplification) of benign and malignant breast lesions, no positive hybridization signal was detected in cancer cells, mammary epithelial cells and stromal lympho-cytes. Using laser capture microdissection and PCR amplification, cancer cells and stromal cells were captured respectively from 12 ca-ses of PCR positive and 12 cases of PCR negative of breast cancer specimens, we found EBV DNA was only amplified in mesenchymal cells. In the detection of the EBER expression with EBV RNA probe and in situ hybridization, the results of 75 cases of benign and malignant breast lesions ( including 12 cases of breast cancer by positive PCR amplification) were all negative. Conclusions The re-sults indicate that the tumorigenesis and development of breast cancer have nothing to do with EBV infection in all cases were chosen.
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Objective To evaluate the effectivness of photodynamic therapy (PDT) with chloroaluminum sulfonated phthalocyanine (CASPc) in B16F10 melanomas in a rabbit model. Methods B16F10 murine melanoma tumor fragments were implanted transclerally into the subchoroidal space of 38 immunosuppressed New Zealand albino rabbits and examined with indirect ophthalmoscopy and ultrasonography. When the tumors ranged from 2.0~3.8 mm in height, 30 rabbits were treated by PDT, with intravenous injection of CASPc 5 mg/kg, and irradiation at the wavelength of 675 nm of an argon-pumped dye laser after 24 hours. Light dose ranged from 20 J/cm 2 to 70 J/cm 2. The other 8 animals were treated with light only or photosensitizer only. The animals were followed up for 6~8 weeks. Results The 30 tumor-bearing rabbits were by PDT (laser and CASPc) regressed in 21 animals after treatment. At light doses under 40 J/cm 2, tumor regrowth was observed in 9 animals after two weeks of treatment. In all of the 8 control animals, the tumor-bearing eyes were filled with tumor at the third week after implantation with laser doses of 70 J/cm 2. Conclusion The study suggest that CASPc PDT may be effective in the treatment of B16F10 choroidal melanomas.