[Establishment of a prediction model for colorectal cancer immune cell infiltration based on the cancer genome atlas (TCGA) database].

OBJECTIVE: To study the correlation between immune cell infiltration in colorectal cancer tissue and clinical prognosis and to explore the levels of some immune cell genes for predicting the prognosis of patients with glioma colorectal cancer. METHODS: In this study, we extracted colorectal cancer data from the cancer genome atlas (TCGA). Based on a deconvolution algorithm (called CIBERSORT) and clinically annotated expression profiles, the analysis assessed the infiltration patterns of 22 immune cells in colorectal cancer tissue to determine the association between each cell type and survival. Differences in five-year survival rate effectively illustrate the clinical prognostic value of each immune cell proportion in colorectal cancer, using a bar graph, correlation-based heatmap to represent the proportion of immune cells in each colorectal cancer sample. RESULTS: A total of 473 colorectal cancer tissues and 41 normal control tissues were extracted from the TCGA database, and the comparative analysis showed that there were differences in the proportion of various TIICs in colorectal cancer tissues, which could characterize individual differences and have prognostic value. Among the cell subsets studied, the proportions of memory B cells, plasma cells, CD4+ T cells, natural killer (NK) cells, M0 macrophages, M2 macrophages, and activated mast cells were significantly different between normal and cancer tissues. Resting NK cells, CD8+ T cells, and plasma cells were associated with T phase, activated dendritic cells were associated with N phase, and eosinophils, M1 macrophages, and activated mast cells were associated with M phase. Survival analysis showed that activated dendritic cells were positively associated with five-year survival rate in colorectal cancer patients. Naive CD4+ T cells were inversely associated with five-year survival rate. CONCLUSION: There are different degrees of immune cell infiltration in colorectal cancer tissues, and these differences may be important determinants of prognosis and treatment response. We conducted a new gene expression-based study of immune cell subtype levels and prognosis in colorectal cancer, which has potential clinical prognostic value in colorectal cancer patients.

Prognostic role of MUC-2 expression in patients with gastric carcinoma: a systematic review and meta-analysis.

OBJECTIVE: This meta-analysis aimed to assess the association of MUC-2 expression with clinicopathological parameters in gastric carcinoma (GC) patients. MATERIALS AND METHODS: Clinical databases based on the study aim were searched in detail. The relative risk ratios (RRs) and associated 95% confidence intervals (95% CIs) were computed after eligible trials were included in the study. RESULTS: Nineteen trials involving 2,363 GC patients were included in this meta-analysis. The expression of MUC-2 showed correlation with clinical stage (I/II vs. III/IV) (RR = 1.09, 95% CI: 1.00-1.18, I2 = 24%, p = 0.194), and lymphatic invasion (present vs. absent) (RR = 0.83, 95% CI: 0.72-0.95, I2 = 22.3%, p = 0.252). However, no significant association was identified between the MUC-2 expression and other clinicopathological parameters, including gender (male vs. female), tumor size (>5 vs. ≤5 cm), Lauren's classification (intestinal vs. diffuse), tumor differentiation (poorly vs. well and moderately), lymph node metastasis (present vs. absent), vascular invasion (present vs. absent), and 5-year survival (yes vs. no) of GC patients. CONCLUSIONS: Our meta-analysis findings suggested that MUC-2 positive cases were correlated with lower tumor stage and lower rate of lymphatic invasion. Further clinical studies are warranted to confirm the role of MUC-2 in clinical practice.

Carfilzomib combined with ex vivo-expanded patient autologous natural killer cells for myeloma immunotherapy.

Natural killer (NK) cell-based immunotherapy is promising, as NK cells are in the first line of defense against cancer and capital of lysing tumor cells without pre-stimulation. However, NK cells from multiple myeloma (MM) patients are always deficient in numbers and the expression of certain activating receptors, disabling them in cytotoxicity against the cancer. Therefore, effective strategies to expand NK cells and increase NK cell-mediated cytotoxicity against MM are significant. Here, NK cells were efficiently expanded from peripheral blood mononuclear cells (PBMCs) of newly diagnosed MM patients after co-culture with irradiated K562 cells transfected with 41BBL and membrane-bound interleukin (IL)-15 (K562-mb15-41BBL) in the presence of 200 IU/ml human IL-2. The ex vivo-expanded NK cells were demonstrated to vigorously kill both MM cells and autologous primary MM cells without significant lysis of patient normal cells. Further exploration revealed a significant increase in cell surface expression of most activating receptors of NK cells and indicated that expanded NK (exp-NK) cell killing of MM cells was mediated by perforin/granzyme. NK cells are capital of lysing human leukocyte antigen (HLA) I-deficient tumor cells and carfizomib, a selective proteasome inhibitor approved for the treatment of relapsed/refractory MM patient, down-regulates the expression of HLA class I, thus enhancing NK cell-mediated lysis in MM. Here, we found for the first time that carfizomib dramatically augmented ex vivo exp-NK cell cytotoxicity against patient autologous MM cells, suggesting the use of exp-NK alone or in combination with the drug to treat MM patient.

[Expression of hypoxia-inducible factor 1α in gastric cancer and its clinical signficance].

OBJECTIVE: To explore the expression level and location of hypoxia-inducible factor 1α (HIF-1α) in gastric cancer (GC) tissues and their relationship with clinicopathological features and clinical outcomes. METHODS: From July to September 2015, 27 pairs of fresh paired GC tissues and adjacent normal tissues were gathered from the Eighth Department of General Surgery of the First Affiliated Hospital of Anhui Medical University. Quantitative real-time PCR (qRT-PCR) and Western blot were performed to detect the expression of HIF-1α mRNA and protein in these tissues. A total of 191 GC tissues and 46 randomly selected adjacent normal gastric tissues were consecutively collected between December 2006 and September 2008 from Department of General Surgery of the same hospital. Immunohistochemistry were performed on them to detect the expression of HIF-1α and CD34[described in terms of microvessel density (MVD)], and correlation of different locations of HIF-1α (in cytoplasm or nucleus) with MVD, clinicopathological features, and clinical prognosis was analyzed. RESULTS: The average relative expression level of HIF-1α mRNA in GC tissues (0.625±0.170) was significantly higher than in normal adjacent tissues (0.218±0.036, t=2.336, P=0.023) by qRT-PCR. From the results of Western blot, the expression level of HIF-1α protein increased in GC tissues compared with its corresponding normal tissues. Immunohistochemistry results revealed that positive HIF-1α staining was observed in 67.54% GC tissues and 45.65% normal tissues, with significant difference (P=0.006). And 35.08% in GC and 45.65% in normal tissues were cytoplasmic positive (P=0.138); while 37.17% in GC and only 2.17% in normal tissues were nuclear positive, with significant difference (P<0.001). High differentiation group and TNM clinical early stage (Ⅰ+ Ⅱ) group had significantly higher cytoplasmic HIF-1α expression positive rate compared with low differentiation group (P=0.008) and TNM clinical intermediate-advanced stage (Ⅲ+ Ⅳ) group (P=0.019); whereas low differentiation group had significantly higher nuclear HIF-1α expression positive rate compared with high differentiation group (P=0.043). The mean MVD in the nuclear HIF-1α positive GC group (115.6 ± 7.8) was higher than that in the cytoplasmic HIF-1α positive GC group (93.1±7.5, t=2.077, P=0.040). The median follow-up time was 56(3-81)months. Kaplan-Meier survival analysis and Log-Rank test results showed that nuclear HIF-1α positive patients had a shorter survival time (median 45 months) than cytoplasmic HIF-1α positive patients (median 64 months, P<0.001). Multivariate Cox regression analysis revealed that differentiation (HR=1.713; 95% CI: 1.019-2.882), depth of invasion (tumor stage, HR=6.137; 95% CI: 1.832-20.556) and lymph node metastasis (HR=2.788; 95% CI: 1.313-5.920) were independent prognostic factors for GC (all P<0.05). CONCLUSION: Different location of HIF-1α protein may be realted to the tumorigenesis and progression of GC, and may become a potential prognostic indicator of GC.

Effects of energy controllable steep pulses on intracellular calcium concentration and cell membrane potential.

BACKGROUND AND OBJECTIVES: Our previous experiments showed that steep pulses could kill tumor cells, but the mechanism is unclear yet. This study was to probe the effects of different dosages of energy controllable steep pulses on intracellular concentration of dissociative calcium ion ([Ca2+]i) and cell membrane potential. MATERIALS AND METHODS: The mammary carcinoma cells MDA-MB-231 were divided into control group and 5 different dosages of Energy Controllable Steep Pulses (ECSP) treatment groups. The calcium ion in each group was labeled by Fluo-3/AM individually and the cell membrane potential was labeled by DiBAC4 (3). The mean fluorescence intensity of fluorescent probe in mammary carcinoma cells was observed in quiet state by laser confocal microscopy after ECSP treatment The changes of calcium concentration and cell membrane potential in cells after ECSP treatment were analyzed. The changes of intracellular [Ca2+]i after ECSP treatment were also observed with and without calcium ion outside of the cells. RESULTS: The calcium ion outside of cells influx with lower dosage of pulse in quiet state. With the dosage increase, the intracellular calcium ion outflow. In real time kinetic detection, the mean fluorescence intensity of intracellular calcium ion was increased with the pulse electric field intensity raised in the lower ECSP. When the voltage was 285V, frequency was 100Hz, the [Ca2+]i decreased. The increase of intracellular calcium ion concentration was decreased without calcium ion than with calcium ion outside of cells, but still raised gradually. The lower dosage of ECSP could induce the fluorescence intensity of DiBAC4 (3) in cells increase, which showed that the lower dosage of ECSP could induce the depolarization of cells. With the dosage raised, the fluorescence intensity of DiBAC4 (3) in cells attenuated. This dosage of ECSP could induce the superpolarization of cell membrane. CONCLUSIONS: The lower dosage of ECSP can induce the depolarization of cell membrane and induce the inter flow of calcium ion outside of cell membrane. The higher dosage of ECSP can directly destroy the cell membrane and induce the superpolarization of cell membrane, then induce the outflow of intracellular calcium ion which causes the necrosis of tumor cells.

Intense nanosecond pulsed electric fields promote cancer cell apoptosis through centrosome-dependent pathway involving reduced level of PLK1.

BACKGROUND: Intense nanosecond pulsed electric fields (nsPEFs) have been known to promote apoptosis without physically changing membrane structure or damaging morphology of tumor cells. To determine the contribution of centrosome to the progression of apoptosis by nsPEFs, HeLa cells were exposed to high intensity (6 kV/cm) nsPEFs (8-32 ns) in normal culture condition and cell biology and molecular parameters of cells were investigated. MATERIALS AND METHODS: Apoptotic cell death was identified by TUNEL assay after being exposed to the nsPEFs with various pulse durations, while immunofluorescent staining was performed to detect the number and distribution of centrosomes. To clarify whether nsPEFs-induced centrosome over-duplication is the consequence of DNA damage, we used comet assay to detect simultaneous DNA damage. And additionally Western Blot was used to detect PLK1 protein level to explore the correlation between apoptotic cell death and nsPEFs-induced centrosome over-duplication. Correlation between nsPEFs and molecular parameters was statistically analyzed. RESULTS: NsPEFs induced a clear apoptosis reaching a maximum at 24ns, 24h after exposure (p < 0.05), where DNA fragmentation and over-duplicated centrosomes were observed. This apoptosis may be promoted in a time- and pulse duration-dependent manner. Polo-like kinase (PLK1) protein levels were significantly decreased by such nsPEFs (p < 0.05). Control treatment without the nsPEFs did not cause any damage to the cultured HeLa cells. CONCLUSIONS: Intense nsPEFs promote cell apoptosis through a centrosome-mediated pathway involving a reduction in the level of PLK1, which may provide new therapeutic targets for human cancer treatment.

High-intensity electric pulses induce mitochondria-dependent apoptosis in ovarian cancer xenograft mice.

Human ovarian cancer models were established in nude mice by transplanting SKOV(3) cells, and then tumors were exposed to high-intensity electric pulses with a voltage 1000 V, frequency of 1000 Hz, and duration of 250 ns for 1 min. Mitochondria permeability transition pore (PTP) was inspected by cofocal microscope; cytochrome C (Cyt C) and apoptosis-induced factor (AIF) were determined by immunohistochemistry; and voltage-dependent anion channel (VDAC) was measured by immunofluorescence. High-intensity electric pulses exposure led to increases of PTP, Cyt C, and AIF and a decrease of VDAC. These findings revealed that high-intensity electric pulses activated mitochondria electroporation, apoptosis was realized via mitochondria pathway.

Retinopathy in a Chinese population with type 2 diabetes: factors affecting the presence of this complication at diagnosis of diabetes.

This study examined the prevalence of retinopathy in 2131 patients with type 2 diabetes attending a Beijing hospital for the first time. The median age of patients was 58 years (IQR 50-65). The overall prevalence of retinopathy was 27.3% (95% CI: 25.4-29.2) and for proliferative retinopathy 7.8% (95% CI: 6.7-8.9). When all patients were considered together, duration of diabetes (OR=1.8; P=0.001) and albumin excretion rate (OR=1.5; P=0.019) were independent risk factors for retinopathy. Blue-collar occupation (OR=1.5; P=0.047) and blood pressure (OR=1.2; P=0.021) were additional risk factors for non-proliferative and proliferative retinopathy respectively. Amongst the 773 newly diagnosed patients, 21% (95% CI: 17.8-23.6) already had retinopathy. The median age of those patients with retinopathy at diagnosis of diabetes was 3 years higher that those without retinopathy, and blue-collar workers (OR=2.2; P=0.012) as well as female gender were particularly at risk (OR=2.0; P=0.033). There was a strong correlation between duration of diabetes with the presence of retinopathy (r=0.95; P=0.01). By extrapolation, it could be estimated that some degree of hyperglycaemia might have been present for more than 20 years before diabetes was diagnosed. These findings emphasise the importance of earlier diagnosis of diabetes and its complications, especially in socially disadvantaged groups.

[Tumor necrosis factor in peritoneal fluid of infertile women with endometriosis and its relation to sperm motility].

Tumor necrosis factor (TNF) of 16 infertile women with endometriosis was measured in their peritoneal fluid (PF) with double monoclonal antibody ELISA method, and its effect on sperm motility and membrane integrity was evaluated by semen analysis and hypoosmotic swelling test (HOS). Comparing with that of infertile women without endometriosis (n = 11) and normal fertile women (n = 7), the level of PF-TNF was significantly higher in women with endometriosis (P < 0.01), in accord with the stage of the disease. In the PF of these patients after incubation with the donor sperms, the percentages of forward movement, total motility and hypoosmotic swelling of the donor sperms were significantly decreased (P < 0.01), showing a negative correlation to TNF levels. These results indicate that the elevation of PF-TNF in endometriosis has detrimental effect on sperm function.