Objective To investigate the significance of abnormal gene in histologically normal mammary epithelial tissue for the early diagnosis of breast cancer. Methods Microarray technology was used to identify abnormal gene expres-sion and analyzed bioinformatics of normal mammary epithelial tissue in breast cancer patients and healthy normal control to establish a model for early diagnosis of breast cancer. The differentially expressed genes were screened by using signal path-way enrichment analysis. The accuracy (Ac), sensitivity (Sn) and specificity (Sp) were used to measure the prediction accura-cy of the different methods. Results The best prediction model was derived from the combination of differential genes en-riched from KEGG and BioCarta database. The number of differential expressed genes in three random created prediction models was reduced from 22 to 7, 14 to 3 and 18 to 4. However, the prediction accuracy was consistent with the model estab-lished from all of the differentially expressed genes, and the average accuracy of all models was 96.3%. Conclusion The prediction model can be simplified with the prediction accuracy unchanged, and thus facilitate the model apply to early diag-nosis and prevention of breast cancer.

Tissue factor pathway inhibitor-2 (TFPI-2), a member of the Kunitz-type family, is a broad-spectrum serine proteinase inhibitor. The expression of TFPI-2 is inversely related to increasing degree of malignancy, suggesting a role of TFPI-2 in the mainte-nance of tumor stability and inhibition of the growth of neoplasma. Aberrant methylation of TFPI-2 promoter cytosine-phosphorothio-ate-guanine (CpG) islands has been widely documented to be responsible for diminished expression of TFPI-2 mRNA and protein dur-ing cancer progression. TFPI-2 expression is significantly up-regulated by the ERK1/2 and JNK signaling pathways and modestly in-creased by VEGF, TNF-alpha, and fibroblast growth factor in time-and dose-dependent manners. TFPI-2 can maintain the stability of the tumor environment and inhibit invasiveness and growth of neoplasms. TFPI-2 has also been shown to regulate proliferation, apopto-sis, and vasculogenic mimicry of tumor cells, which may contribute significantly to tumor growth inhibition. Restoration of TFPI-2 ex-pression in tumor tissue inhibits tumor growth and metastasis, which creates a novel possibility of cancer patient treatment. This review focuses on the expression and the molecular regulation mechanisms of TFPI-2 in malignant tumors that control the functions of TFPI-2 in proliferation, apoptosis, and angiogenesis. Insight into these processes will improve our understanding of TFPI-2 and provide new ap-proaches for rational treatment strategies.

Objective:To study the direct effects of gradient concentration inteiferon-a(IFN-a) on lymphoma cells line Daudi and Jur-kat and a group of 15 cases refractory lymphoma. Methods: The effects of IFN-a on the growth of tumor cells were assayed by MTT, apoptosis were measured by flow cytometty, TUNEL and electron-microscope, 15 refractory lymphoma cases were treated by local injection of IFN-a combination chemotherapy.Results: Low-dose IFN-a did not have any significant effects on growth of Daudi and Jurkat cells, high-dose IFN-a (10 000U/ml) not only have significant anti-proliferative effects with dependence of time and dosage but also can induce apoptosis on both Daudi and Jurkat cells. 5 patients achieved a complete remission and 7 patients achieved a partial remission, the overall response rate was 80% .Conclusion:IFN-a( 10 000 U/ml) had significant anti-proliferative effects and inducing apoptosis on lymphoma cells.Local injection of IFN-a is one of effective therapy on refractory lymphoma.

Purpose:To analyze the immunophenotype character of malignant lymphoma with bone marrow involvement by means of Flow Cytometry (CD45/SSC gate).Methods:Bone marrow of malignant lymphoma patients was detected by Flow Cytometer(CD45/SSC gate),bone marrow smear was simultaneously performed as control.Results:(1) Bone marrow of 34 malignant lymphoma patients was examined by Flow Cytometry.23 cases were detected to have bone marrow involvement.(2) Among these 23 cases,19 cases were non-Hodgkin's lymphoma(NHL),4 cases were Hodgkin's lymphoma(HL).The highest frequency antigen marker of B cell NHL was CD19 and CD20,T cell NHL was CD7,and HL was CD9.Conclusions:(1)Flow Cytometry(CD45/SSC gate) is a feasible and effective method to detect patients with bone marrow involvement.(2) The antigen marker of B cell NHL is CD19 and CD20,T cell NHL is CD7 and HL is CD9.

Objective: To explore the effects of various concentrations IFN-? on human oral epidermic cancer cell line KB and its multidrug resistant counterpart KBv200.Methods: The toxicity of IFN-? and/or VCR was assayed by MTT method; intracellular rhodamin123 concentration and p-glycoprotein expression were measured by flow cytometry; mdr-1 mRNA was assayed by RT-PCR. Results: High-dose IFN-?(10 000 IU/ml) had significant antiproliferative effects on KB/KBv200 cell lines. Low-dose IFN-? did not have any significant effect on growth of KB/KBv200 cells, but did increase the cytotoxic effect of VCR and the accumulation of Rhodamine123 in KBv200 cell line and had no effect on parent VCR-sensitive KB cell line. IFN-? down-regulated expression of P-gp and mdr-1mRNA . Conclusions: IFN-? has significant effect on the reversal of multidrug resistance of KBv200 cell line via a mechanism of P-gp and mdr1 mRNA down-regulation.

Objective:To observe the clinicopathological features of gastrointestinal stromal tumor (GIST) cases with concurrent carcinoma. Methods:Patient data of 24 GIST cases with concurrent carcinoma were collected from the 157 GIST cases reported be-tween 2002 and 2012. The clinicopathological features of the GIST cases with concomitant carcinoma were studied. The expression of CD117, CD34, and SMA by the tumors was assayed using the immunohistochemical EliVision method. In particular, the expression of the proliferation marker Ki-67 was studied. Results:GIST cases with concurrent carcinoma accounted for 15.3%of the total GIST cas-es studied. The GIST patients with concurrent carcinoma included 14 males and 10 females. The male-female ratio of these patients was 1.4∶1. The age of the patients ranged from 41 years to 66 years, with a median age of 55 years. Lesions at the inferior segment of the esophagus were found in 7 of the 24 selected GIST cases;lesions at the gastric wall and in the intestines were observed in 15 and 2 cas-es, respectively. The diameter of the GIST cases with concurrent carcinoma ranged between 0.6 and 3.8 cm, with an average of 1.50 ± 0.85 cm. Slight dysplasia was observed in 4 of the 24 cases; no heteromorphism was present in the remaining 20 cases. The mitotic counts of GIST cases with concurrent carcinoma ranged from 0/50 HPF to 5/50 HPF, with an average of (0.79±1.83)/50 HPF. The pro-liferative index of Ki-67 in the GIST cases with concurrent carcinoma ranged between 0 and 7.72, with an average of 2.16 ± 3.26. The concurrent carcinoma cases included 5 cases with esophageal carcinoma, 2 with cardiac carcinoma, 15 with gastric cancer, and 2 with intestinal cancer. In contrast to the GIST cases with concurrent carcinoma, the GIST cases without carcinoma complications included 74 males and 59 females. The male-female ratio was 1.25∶1. The age of the patients without concurrent carcinoma ranged from 43 years to 71 years, with a median age of 54 years. Among the 133 GIST cases without cancer complications, gastric, intestinal, and esophageal lesions were found in 114, 13, and 6 cases, respectively. The diameter of GISTs without cancerous complications ranged from 2.4 cm to 15.5 cm, with an average of 6.11 ± 7.09 cm. Different degrees of dysplasia were seen in 82 of the 133 cases. The mitotic counts in the GIST cases without cancer complications ranged from 0/50 HPF to 53/50 HPF, with an average of (3.81±23.67)/50 HPF. The prolifera-tive index of Ki-67 for these cases ranged from 0 to 39.21 and averaged at 6.22 ± 16.96. The male-female ratio of the GIST cases with cancer complications was higher compared with the GIST cases without. The average diameter of GISTs with complications was small-er compared with that of GISTs without complications. The mitotic counts and the proliferative index of Ki-67 were significantly lower in the GIST cases with cancer complications than in those without (t=1.981, P<0.05 vs. t=1.993 5, P<0.05). Conclusion:Concurrent car-cinomas were found in 15.3% of the total GIST cases. No special clinical symptoms were observed in most GIST cases with cancer complications, as revealed when the carcinomas were examined. The proliferative index of Ki-67 in the GIST cases with concurrent car-cinoma is significantly lower compared with that of the GIST cases without complications.

Objective: This study aimed to examine the number of activated circulating endothelial cells (aCECs) in the peripheral blood of patients with non-small cell lung cancer (NSCLC), and investigate the relationship among aCECs, anti-angiogenic therapy, and prognosis of NSCLC patients. This study also aimed to identify novel predictive markers for anti-angiogenic therapy, and provide basic data and experimental basis for establishing an evaluation system for this therapy. Methods: A total of 142 NSCLC patients were randomly divided into the chemotherapy group (Group 1) and combined therapy group (i.e., chemotherapy plus endostatin, Group 2). The number of aCECs was measured using flow cytometry by detecting the expression status of CD105 and CD146 in the peripheral blood. The correlation between the changes in aCECs and efficacy of drug treatment was statistically analyzed using SPSS software. Results:The number of aCECs in Group 2 increased significantly at 8 and 29 d, two cycles, 50 and 71 d, and four cycles after treatment, respectively (P<0.05). In particular, aCECs amount in cases of progressive disease increased more significantly after combined therapy (P<0.05). A negative correlation was found between the treatment cycle and difference in aCECs amount before and after therapy (r=-0.970, P=0.001). A negative correlation was also observed between the difference in aCECs amount and time to tumor progression (TTP) (r=-0.351, P=0.039). Therefore, the difference in aCECs amount before and after therapy could serve as an important predictor for TTP in NSCLC patients. Conclusion:CD105 and CD146 reflected the activation status of endothelial cells, and responded to the drug treatment. Thus, CD105 and CD146 could act as ideal markers for aCECs. The number of aCECs increased during cancer progression, but significantly decreased after long-term treatment. Therefore, the change in aCECs amount may be a useful marker in predicting the efficacy of anti-angiogenic therapy.

Objective:To highlight the developmental process of 3D cell culture technology system, which is more suitable for isolating and identifying lung cancer stem-like cells than 2D cell culture technology system, and to explore the application of 3D cell cultures in the evaluation of proliferation, apoptosis, invasion, and drug resistance of lung cancer. Methods:Cells (104/well) from the human lung adenocarcinoma cell lines A549 and RPMI 1640 were cultured in complete medium containing 10%fetal bovine serum. Cell suspension was cultured in a BME basal medium. A growth curve was drawn after 7 d of culture. The stem-like cell was identified through a mammosphere culture, drug resistance and invasion assay, and flow cytometry. Data of A549 cells cultured in 3D and 2D tra-ditional cell culture technologies were compared. Results: Cells from the 3D cell culture had higher tumor formation rates [(20.75 ± 0.85) d vs. (60.25 ± 1.49) d, P<0.01)] and tumor sphere formation (28.50%± 1.17%vs. 8.67%± 0.80%, P<0.01) than those from the 2D cell culture. Moreover, cells from 3D cell culture were more invasive and resistant to therapy (58.17%± 2.19%vs. 41.70%±5.81%in 48 h, P<0.01;33.27%±5.76%vs. 27.30%±4.25%in 72 h, P<0.01). Phenotype experimental results demonstrated that the CD44 and CD326 cells were double-positive, whereas the CD24 cell was negative. Conclusion:The proportion of stem-like cells in A549 cell line after 3D cell culture significantly increased compared with 2D cell culture. The 3D cell culture can promote the proliferation of lung cancer stem cells.

OBJECTIVETo investigate the expression of apoptotic regulator c-FLIP(L) in invasive breast carcinoma tissues, and to evaluate its correlation with molecular subtyping and clinical prognosis.

METHODSImmunohistochemistry using EnVision staining for c-FLIP(L) was performed in 264 cases of invasive breast carcinomas and matched adjacent normal breast tissue samples from January 1996 to December 1999. ER, PR, HER2, Ki-67, CK5/6 and EGFR were evaluated by immunohistochemistry in order to classify the tumors into five molecular subtypes and the difference of c-FLIP(L) expression in these molecular subtypes was also analyzed. The influence of c-FLIP(L) expression on prognosis was evaluated by Kaplan-Meier curves and multi-factor Cox proportional risk model.

RESULTSHigh expression of c-FLIP(L) was observed in 84.5% (223/264) of cases of invasive breast carcinomas which were significantly higher than the 45.1% (119/264) of cases in adjacent normal epithelium of breast (χ² = 89.78, P = 0.000). The expression of c-FLIP(L) in luminal B (HER2 positive) and basal-like breast cancers was 78.1% (25/32) and 46.2% (18/39), respectively, with significant difference (P < 0.05). Moreover, the expression of c-FLIP(L) in luminal B (HER2 positive) was higher than in luminal A cancers (P < 0.05), and the expression of c-FLIP(L) in HER2 positive cancers was higher than in basal-like cancers (P < 0.01). C-FLIP(L) showed deep yellow staining in node positive breast cancer with a high-expression rate of 93.1% (134/144); whereas the expression was sporadic and light yellow in node negative breast cancer with a lower high-expressed rate of 72.5% (87/120, P < 0.01). C-FLIP(L) expression had significant influence on disease-free survival time, with c-FLIP(L)-positive patients showing poor prognosis (P < 0.01). Multi-factor Cox proportional risk model analysis showed that expression of c-FLIP(L), lymph nodes status and molecular subtypes were independent prognostic factors for invasive breast carcinomas (P < 0.05).

CONCLUSIONSC-FLIP(L) is highly expressed in invasive breast carcinomas, and its expression level is closely related to the molecular subtypes and clinical prognosis of breast cancer patients. Thus, c-FLIP(L) could be used as an important tumor marker for personalized cancer therapy and prognostic prediction.

Aged ; Biomarkers, Tumor ; metabolism ; Breast ; metabolism ; Breast Neoplasms ; classification ; metabolism ; mortality ; CASP8 and FADD-Like Apoptosis Regulating Protein ; metabolism ; Disease-Free Survival ; Female ; Humans ; Immunohistochemistry ; Prognosis ; Receptor, ErbB-2 ; metabolism

Objective: To verify the safety and efficacy of IONTRIS particle therapy system (IONTRIS) in clinical implementation. Methods: Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial: 31 males and 4 females with a median age of 69 yrs (range 39-80). Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non-metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results: Twenty-two patients received carbon ion and 13 had proton irradiation. With a median follow-up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression-free survival rate was 93.8% (15/16). Among the 19 patients with prostate cancer, biological-recurrence free survival was 100%. Particle therapy was well tolerated in all 35 patients. Twenty-five patients (71.4%) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow-up. Six (17.1%) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow-up is needed to observe the late toxicity and long term result.