Objective To observe EML4-ALK fusion gene mutation expression rate in serum and cancer tissue of patients with non-small cell lung cancer (NSCLC) in Chinese populations,and the consistency of mutation in serum and cancer tissues,and the feasibility of real-time,dynamic detection of EML4-ALK fusion gene therapy by using FQ-PCR.Methods 123 cases of serum and 98 cases of tissue of NSCLC patients were detected by fluorescence quantitative polymerase chain reaction,and 77 cases of which were matched.The clinical curative effects of ALK inhibitor (crizotinib) were analyzed.Results 13 rearrangement in 123 (10.6％) of the patients'serum samples and 11 rearrangement in 98 (11.2％) tumor tissue.EML4-ALK rearrangement were mainly discovered in adenocarcinoma (x2 =4.083,P =0.036),and non-smokers in NSCLC (x2 =5.326,P =0.019).The consistency of patients with EML4-ALK matched tumor tissue and serum reached 66.7％ (6/9,κ =0.779).The EML4-ALK fusion gene rearrangement in patients receiving ALK inhibitor (crizotinib) treatment achieved significant benefit.Conclusion The EML4-ALK rearrangement mainly exists in the serum and tumor tissue of adenocarcinoma and non-smokers in NSCLC.When tumor tissue samples are unable to be obtained,FQ-PCR can be used to detect serum EML4-ALK fusion gene mutation for selecting NSCLC patients suitable for crizotinib therapy instead of tumor tissue.

Her-2( human epidermal growth fact or receptor 2) is a type of 185 kD across the membrane glycoprotein with tyrosine kinase activity,play an important function in the cell divisions across a cell membrane proliferation signal transduction,and finally affect biological activities of tumor cells from multiple ways,such as a tumor cell proliferation and adhesion,transfer and differentiation and other related gene regulation.Recent studies show that Her-2 overexpression rate of gastric cancer patients is about 12％ ～ 35％,and Her-2 is a gastric cancer poor-prognostic factors,while Her-2 monoclonal antibody-Trastuzumab is expected to become a new standard gastric cancer treatment for the patients with Her-2 overexpressed.

Objective To examine the positive rate of c-MET gene amplification in primary and lymph node-metastatic non-small cell lung cancer( NSCLC),and to explore their relationships. Methods From November 2011 to November 2013,147 cases of primary NSCLC consisting of 71 cases of paired lymph node-metastatic tumors and 47 cases of normal lung specimens as the control group were collected in General Hospital of Beijing Military Region. The c-MET gene copy number was examined by RT-PCR and the positive rate of c-MET gene amplification among NSCLC population was figured out,thus the consistency of c-MET gene ampli-fication in advanced primary NSCLC and associated lymph node-metastases and the relationship between c-MET gene amplification and clinical data were analyzed. Results The positive rate of c-MET gene amplification on primary tumor was 8. 84% (13 / 147). For those 71 paired cases,the positive rate on primary tumor was 8. 45%(6 / 71),with that of lymph node-metastases 18. 31%(13 / 71). Among the 71 cases,there were 8 cases whose metastases were positive but primary tumors negative and 1 case whose primary tumor was positive but metastases negative. It was of statistical significance between the two groups(McNemar test,χ2 = 4. 274, P = 0. 039). The positive rate of primary tumors could be predicted by lymph node-metastases(κ = 0. 464, P ﹤ 0. 001). The sensitivity was 83. 3% and the specificity was 87. 7% . Positive rate of c-MET amplification was higher in male and smoking patients with lymph node-metastases above N2 . Conclusion c-MET amplifica-tion test should be one of the routine genetic testing projects. The amplification on primary tumors is higher than that on lymph node-metastases,implying that metastases test can pick out more patients with indication. Metas-tases test can predict the amplification on primary focus,and it is an alternative way to guide the treatment of c-MET target medicine. Moreover,the clinical characteristic can be served as an indicator of positive c-MET am-plification.

Pulmonary embolism(PE)is a relatively common cardiovascular emergency.Haemodynamic instability is the characteristics of massive pulmonary embolism,with a mortality as high as 20%.The goals of PE treatment are to remove the thromboembolic obstruction rapidly,exert beneficial effects on haemodynamic parameters and save the lives of patients.Haemodynamic and respiratory support is of vital importance in some critically ill patients with PE.The basic treatment is anticoagulation,the methods of which are different for PE during pregnancy,among cancer patients,with right heart thrombi and with heparin-induced thrombocytopenia.

0.05), but it began to decrease ( P

AIM: To study the effect of interleukin-1β (IL-1β) on the activity of vacuolar proton pump (V-ATPase) and intracellular pH value in the osteoclasts for exploring the mechanism of IL-1β to promote osteoclastic bone absorption.METHODS: The mature osteoclasts were attained by induction of bone marrow monocytes/macrophages.The osteoclasts were cultured with α-MEM and treated with different concentrations (0.1, 0.3 and 0.5 μg/L) of IL-1β.After cultured for 48 h, the mRNA and protein expression of V-ATPase was detected.The effect of IL-1β on intracellular pH value, activity of V-ATPase and the bone absorption abilities of osteoclasts were examined.RESULTS: The expression level and activity of V-ATPase were significantly increased and the intracellular pH value of the osteoclasts decreased after incubated with IL-1β for 48 h.Under the same culture condition, the bone absorption capacity of the osteoclasts was promoted.The enhanced bone absorption capability of the osteoclasts was accompanied by an increase in the concentration of IL-1β.CONCLUSION: The mechanism of IL-1β participating in pathological bone absorption in the bone and joint inflammatory diseases is that IL-1β increases the expression and activity of V-ATPase, boosts the production and/or transportation of hydrogen ion, leading to increased osteoclastic bone absorption activity, and resulting in bone destruction.

ObjectiveTo investigate the expression of urotensin Ⅱ(UⅡ) in the lung cancer tissue from surgical resection of lung cancer patients,and to detect the relationship between UⅡ expression and pathologie types and the clinical stages of lung cancer.MethodsThe expression rates of UⅡof 45 lung cancer tissues and 20 inflammatory pseudotumor were measured by immunohistochemical assay,and the relationship between UⅡ expression and the pathologic types and clinical stages of lung cancer was analyzed.ResultsUⅡwas mainly distributed in lung cancer cell cytoplasm,which was tan-yellow particles.The positive expression rate of UⅡin nonsmall cell lung cancer was 61.3％ (19/31),which was higher than that in small cell lung cancer(7.1％,1/14)and pulmonary inflammatory pseudotumor( 15.0％,3/20) (P ＜ 0.01 ).The positive expression rate of UⅡ was 100％ in adenocareinoma.The positive expression rate of UⅡin staging Ⅲ non-small cell lung cancer( 85.7％ )was higher than that of staging Ⅰ ( 16.7％ ) ( P ＜ 0.05).ConclusionUⅡ cxists in the cytoplasm of lung cancer cells,and the expression of UⅡis correlated with the pathological type and TNM staging of lung cancer.

BACKGROUND:Previous studies have applied long-bone mechanical separation method to obtain osteoclasts, which are terminal y differentiated cells and cannot further proliferate. Therefore a large number of osteoclasts can be harvested with bone marrow cells inducing culture method and RAW264.7 cells inducing culture method to meet clinical requirements.
OBJECTIVE:To investigate the optimal method of receptor activator of nuclear factor kappa-B ligand (RANKL) induced osteoclast precursors to differentiate into mature osteoclasts.
METHODS:After bone marrow cells were isolated from mouse, RANKL and macrophage colony stimulating factor were added into the medium together, or RAW264.7 cells were cultured with RANKL to induce osteoclasts. The osteoclast precursors were treated with different concentrations of RANKL to observe the number of generated osteoclasts and evaluate the dose-effect relationship between RANKL and osteoclastogenesis. Annexin V-FITC and propidium iodide staining were used for flow cytometry to analyze the mononuclear-macrophage apoptosis during osteoclastogenesis.
RESULTS AND CONCLUSION:When 10μg/L RANKL was used, the peak of osteoclastogenesis appeared at days 6-7;when the concentration of RANKL was up to 100μg/L, the peak appeared at days 4-5. The number of new osteoclasts was dose-dependent on the RANKL concentration. 50μg/L of RANKL was the turning point in the fitted curve from osteoclastogenesis and RANKL concentration. After the RANKL concentration was more than 150μg/L, the number of osteoclasts slowed down obviously. RANKL can induce monocyte-macrophage to form osteoclasts and promote monocyte-macrophage apoptosis. The highest number of osteoclasts would be obtained to each unit of RANKL when monocyte-macrophage cells were cultured with 30μg/L of RANKL in the same vaccination density (104/cm2). Experimental findings indicate that, RAW264.7 cells or bone marrow cells inducing culture methods are simple and feasible, the optimum cellseeding density was 104/cm2;the appropriate stimulation concentration of RANKL was 30-50μg/L.

Objective　To explore the effects of hypoxia on the syntheses and secretion of adrenomedullin (AM), calcitonin gene related peptide (CGRP) and c-type natriuretic peptide (CNP) and the relationship between these peptides. Methods　Rat models were established with hypoxia for 10, 20 and 30 d respectively and rats under normal altitude were served as control. Pulmonary artery pressure and the maximum increasing speed of right ventricle (RVdp/dtmax) were measured in every group. The dynamic changes of AM, CGRP and CNP concentrations in plasma were studied with radioimmunoassay. Results　During hypoxia, pulmonary artery pressure and RVdp/dtmax were enhanced. Plasma AM and CNP concentrations were increased while CGRP was decreased significantly. The plasma level of AM had positive correlation with that of CNP, but negatively correlated with that of CGRP. Conclusion　Results indicate that hypoxia may cause pulmonary artery pressure change and right ventricle has compensatory reaction to hypoxic pulmonary hypertension. Dynamic changes of plasma AM, CGRP and CNP concentrations can be regarded as indexes for condition of illness.

Objective:To evaluate the efficacy and safety of combined cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) with lobaplatin and docetaxel for treatment of synchronous peritoneal carcinomatosis (PC) from gastric cancer. Methods:Fifty patients with synchronous PC from gastric cancer were treated by 52 CRS+HIPEC procedures with 100 mg of lobaplatin and 120 mg of docetaxel in 12000 mL of normal saline at (43 ± 0.5)℃for 60 min. The primary and secondary endpoints were overall survival (OS) and perioperative safety profiles, respectively. Results:At a median follow-up of 22.5 months, the median OS rate was 14.3 (95%CI:7.6-21.0) months, and the 1-, 2-, and 3-year survival rates were 58%, 40%, and 32%, respectively. No perioperative deaths or serious adverse events occurred in 12 cases (23.1%). Multivariate analysis indicated that completeness of cytoreduction, nor-mality of perioperative tumor markers, and adjuvant chemotherapy of more than six cycles were independent predictors for improved survival. Conclusions:CRS+HIPEC with lobaplatin and docetaxel could improve the OS and ensure perioperative safety of patients with synchronous PC from gastric cancer.