Genomic mutation characteristics and prognosis of biliary tract cancer.

BACKGROUND: The incidence of biliary system cancer is higher in the Chinese population than in the West. The overall prognosis of gallbladder cancer and cholangiocarcinoma is poor, and the current treatment is limited. In order to explore the pathogenesis of biliary tract cancers and potential targeted therapies, we mapped the mutation landscape of biliary tract cancer in the Chinese population and analyzed the molecular mechanism related to prognosis. METHODS: A total of 59 formalin fixed paraffin-embedded (FFPE) tissue samples were obtained from patients with operable biliary tract cancer. We conducted targeted capture sequencing of 620 genes through high-throughput sequencing technology and analyzed the fusion information of 13 genes. RESULTS: Mutations were detected in 88% samples, and the most frequent mutation base was C>T. Genes with higher single nucleotide variations (SNV) and copy number variations (CNV) frequency are TP53, KRAS, ARID1A, VEGFA, cyclin family related genes and cyclin-dependent kinase genes. Actionable mutations were detected in 59.3% samples, and germline mutations were detected in 22% samples. Patients with KRAS mutations, VEGFA pathway mutations and higher tumor mutation burden (TMB) may have poor prognosis. CONCLUSIONS: We explored the mutation characteristics and prognostic mechanism of biliary tract cancers in the Chinese population. This study provides potential evidence for targeted therapy and immunotherapy of biliary tract cancers.

Changes and Clinical Significance of Detailed Peripheral Lymphocyte Subsets in Evaluating the Immunity for Cancer Patients.

OBJECTIVE: The evaluation of lymphocyte subsets is widely regarded as an important factor for monitoring tumor progression and response to therapy. This study was designed to establish a comprehensive and detailed assessment of peripheral lymphocyte subsets with a multi-parametric flow cytometry assay for response prediction and prognosis evaluation of cancer patients. METHODS: Peripheral blood samples collected from 40 cancer patients and 23 age- and sex-matched healthy volunteers were tested for 29 lymphocyte subsets by flow cytometry. The univariate analysis was applied to establish the reference interval of healthy samples, and the ratio and proportion of 29 lymphocyte subsets between patient samples and healthy controls were compared to evaluate their clinical significance by Mann-Whitney U-test model. RESULTS: The reference ranges of 29 lymphocyte subsets were established with a normal distribution and no significant differences were observed between genders. Compared with healthy control group, lower proportion and ratio of specific parameters, such as Naïve Th cells (p<0.01), Naïve Tc cells (p<0.01), CM (central memory) Tc cells (p<0.01), Naïve T cells/Memory T cells (p<0.001), Naïve T cells/EM (effector memory) T cells (p<0.001) and Naive Th cells/Memory Th cells (p< 0.001), and higher proportion and ratio of EM Th cells (p<0.001), EM Tc cells (p<0.01), effector Tc cells (p<0.05), EM Th cells/CM Th cells (p<0.01) and EM Tc cells/CM Tc cells (p<0.01), as well as Breg (p<0.001), B cells (p<0.05) and CD16-NK cells (p<0.001) were found in cancer cohorts. CONCLUSION: This study suggests that the changes in certain lymphocyte subsets might be helpful to evaluate the immunity of cancer patients, and holds great potential for clinical application.

Activation or suppression of the immune response mediators in biliary tract cancer (BTC) patients: a systematic review and meta-analysis.

Background: Infiltration of immune cells and immune microenvironment determine the proliferative activity of the tumor and metastasis. The aim of this study was to analyze the influence of activation or suppression of the immune response mediators on the prognosis of biliary tract cancer (BTC). Methods: We searched Pubmed, Web of Science, Embase and The Cochrane Library for relevant literatures until June 2016. The quality of studies was assessed by QUADAS-2 and NOS tools. Forest and funnel plots and all statistical analyses were generated by using Review Manager 5.3. The bias of included studies was estimated by Egger's test using Meta R package. Results: A total of 2339 patients from 12 studies were finally enrolled in this meta-analysis. Patients with high expression of immune active factors, intraepithelial tumor-infiltrating CD4+ , CD8+, and Foxp3+ T lymphocytes, MHC I, NKG2D, showed a better overall survival (OS) than those with low expression (HR=0.52, 95% CI=0.41-0.67, P<0.00001). On the contrary, the high expression of immune suppressive factors (CD66b+ neutrophils, Neutrophil-lymphocyte ratio, Intratumoral IL-17+ cells and PD-1+/CD8+ TILs) was significantly associated with poor OS (HR=1.79, 95% CI=1.44-2.22, P<0.00001). A further analysis of therapies targeting tumor microenvironment modulation showed that the median progression free survival (PFS) for BTC patients who received adjuvant immunotherapy was longer than those who received surgery or chemotherapy alone, and the estimated pooled mean difference demonstrated a highly significant improvement (MD =2.33; 95% CI: 0.63-4.02, P=0.007). The total effect of PFS and OS was statistically longer in experimental group, compared to patients in control groups, respectively (PFS: RR=1.25; 95% CI: 1.08-1.46, P=0.004; OS: RR=1.16; 95% CI: 1.07-1.27, P=0.0006). In subgroup meta-analysis of studies on 6-, 12- and 18-month PFS and OS, it showed that adjuvant immunotherapy could improve the 6-month PFS (RR=1.23; 95% CI: 1.05-1.44, P=0.009), and 6-month OS (RR=1.17; 95% CI: 1.06-1.30, P=0.002). Conclusions: So given the above issue, our meta-analysis confirmed that the level of immune mediators could be a predicative factor for prognosis of BTC patients, and immunotherapy regimens by modulating the tumor microenvironment was superior for enhancing median PFS, 6-month PFS and OS.

The combination of dendritic cells-cytotoxic T lymphocytes/cytokine-induced killer (DC-CTL/CIK) therapy exerts immune and clinical responses in patients with malignant tumors.

BACKGROUND: The clinical trials using immunotherapy have been performed for the treatment of variety of malignant tumors. However, large-scale meta-analysis of combined DC-CTL/CIK therapy on immune and clinical response in patients has not been well studied yet. The purpose of this study is to investigate the role of DC-CTL/CIK therapy and evaluate the changes of immune indicators and tumor serological markers both at an individual level and at a system level, which is an important basis for immunotherapy as well as prognosis estimation. METHODS: Three cohorts were designed to estimate therapeutic effects on patients with malignant tumors. Tumor serological markers were detected pre- and post-treatment by immunoradiometric methods using commercially available diagnostic kits. Lymphocyte subsets were identified by flow cytometry. The quality of life was assessed by EORTC QLQ-C30 questionnaire. RESULTS: In this study, we found out that Tregs was significantly reduced after transfusion of DC-CTL/CIK cells companied by decreasing serological tumor markers including AFP, CA199 and CA242 in primary liver cancer and CA724 in gastric cancer. A system-level analysis showed that lower percentages of Tregs were detected in patients with long-lasting courses of immunotherapy. Strikingly, a tumor progression indicator, myeloid-derived suppressor cells (MDSC), was dramatically decreased in patients after DC-CTL/CIK treatment. These results suggested that DC-CTL/CIK therapy improves immune functions and the quality of life post-treatment versus pre-therapy, indicating that DC-CTL/CIK therapy might block the deterioration of invasive cancers in these patients. CONCLUSIONS: This study demonstrated that DC-CTL/CIK therapy could reduce Tregs, MDSCs, and several crucial serological tumor markers in particular tumors, and improve the function of T cells immune systems and the quality of life in patients with malignant tumor.

The combination of an oxygen-dependent degradation domain-regulated adenovirus expressing the chemokine RANTES/CCL5 and NK-92 cells exerts enhanced antitumor activity in hepatocellular carcinoma.

Oncolytic adenoviruses are modified based on adenovirus serotype 5 (Ad5), which belongs to subgroup C and depends on Coxsackie-adenovirus receptor (CAR) to recognize target cells. However, expression of CAR is generally low or lost in certain tumors including hepatocellular carcinoma (HCC). By contrast, CD46 is highly expressed in various types of malignant tumor cells. Therefore, we constructed an adenovirus vector expressing the human RANTES/CCL5 gene regulated by oxygen-dependent degradation domain (ODD) and analyzed its antitumor effects in vitro and in vivo. The human RANTES/CCL5 gene was fused with ODD by PCR and the recombinant oncolytic adenovirus containing RANTES-ODD, SG511-CCL5-ODD, was constructed by the Gateway system, which infected cells by binding CD46. Viral replication experiments were performed to evaluate the selective replication ability of SG511-CCL5-ODD. RANTES expression was determined by ELISA. The chemotactic test was used to analyze the ability of the expressed RANTES to recruit NK92 cells. The antitumor effects of SG511-CCL5-ODD were examined in HCC xenografts in nude mice. A chimeric oncolytic adenovirus, SG511-CCL5-ODD, was constructed successfully. Cells infected with the recombinant virus were able to express RANTES selectively in different environments controlled by ODD and the expressed RANTES was able to recruit NK92 cells by its chemotactic effect in vitro and improve the anticancer immune response in HCC xenografts in nude mice. The chimeric adenovirus SG511-CCL5-ODD highly expressed the RANTES-ODD fusion gene in the hypoxia of HCC under the control of the ODD and effectively attracted NK92 cells and a high number of immunocytes. These factors had complementary advantages and, in combination, exerted enhanced antitumor efficacy.

Effect of moisture transport on microclimate under T-shirts.

Water transport through garments has influence on the microclimate between the garments and the body beneath; thus the thermal comfort feeling for the wearer. Soybean protein fiber (SPF), a new type environmental fiber, which has been reported to be superior in water transfer, is often blended with cotton to improve the water transport property. In this paper, T-shirts made of this SPF/cotton blended fabric were focused in comparison with T-shirts made of cotton fabric. Wicking and immersion tests were carried out on the two types of fabrics to investigate the water transport and absorption properties, respectively; wear trials of T-shirts made of the fabrics were also conducted. Comparing with the cotton fabric which had better water absorptive property, it was found that the blended fabric with superior wicking ability could not only delay the increase of the vapor pressure under the T-shirt at the beginning of the exercise, but also help to keep it lower through the exercise significantly, and also kept the skin temperature under the T-shirt lower. It was made clear that it is the water transfer property rather than the water absorption property helps to take away sweat quickly and prevents the increase of the humidity and temperature at skin surface, thus maintaining a comfort microclimate under garments.

Slow drag in polydisperse granular mixtures under high pressure.

The behavior of polydisperse granular materials, composed of mixtures of particles of different sizes, is studied under conditions of high pressure and confinement. Two types of experiments are performed. In the first type, granular mixtures are compressed, with the resulting force-displacement curve used to calculate density and volume modulus. In the second set of experiments, the drag force is measured by pulling a cylinder, horizontally, through a compressed granular mixture. The density, volume modulus, and drag forces for the mixtures are quantified in terms of the mixture composition. The results show that the behavior of these mixtures depends strongly on the mass fractions of the different sized particles, with density, volume modulus, and drag force all reaching values significantly higher than observed in the monodisperse granular materials. Furthermore, the trends for density and drag force show strong correlation, suggesting that drag resistance of confined granular media could be directly related to packing effects. These results should prove useful in understanding the physics of drag in granular materials under high pressure, such as ballistic penetration of soils or ceramic armors.

Slow drag in granular materials under high pressure.

The resistance offered by a cylindrical rod to creeping cross flow of granular materials under pressure is investigated. The experimental system consists of a confined bed of granular particles, which are compacted under high pressure to consolidate the granular medium. A cylindrical rod is pulled at a constant and slow rate through the granular medium, and the measured pulling resistance is characterized as a drag force. The influence of various parameters such as the velocity of the cylindrical rod, the rod diameter and length, the granular particle size, and the compaction pressure on the required drag force is investigated experimentally. Nondimensional analysis is performed to simplify the relationships between these variables. The results show that the drag force is independent of the drag velocity, is linearly proportional to compaction pressure and rod diameter, and increases with rod length and particle size. Additional compaction experiments show that the effective density of the granular bed increases linearly with pressure, and similar behavior is noted for all particle sizes. These results should prove useful in the development of constitutive equations that can describe the motion of solid objects through compacted granular media under high pressure, such as during ballistic penetration of soils or ceramic armors.