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Cervical cancer is the most common tumor in gynecologic oncology, and the generation and progression of tumor is always associated with immunosuppression. In tumour immunity, T cell-mediated immune response plays a key role.The second signal pathway in T cell activation requires the combination of costimulatory and its receptors. Lacking of the second signal of costimulatory, antigen complexes formed in the first signal pathway can lead to inactivation or dysfunction of T cells. In recent years, B7 family has become an intense focus of research in costimulatory molecules, and can play a positive and/or negative regulatory role in T cell-mediated immune response. This paper is aimed to review the expression and significance of negative costimulatory molecule B7-H4 in the blood and tissues of patients with cervical cancer, and to discuss the problems in the study of cervical cancer, and to introduce briefly the expression and significance in ovarian cancer, as well as to prospect the prognosis, the help of auxiliary diagnosis and immunization therapy. Also it is aimed to further clarify the mechanism of immunosuppression of cervical cancer, and further explore the effective methods of immune targeted treatment. [ABSTRACT FROM AUTHOR]

Introduction. Solitary fibrous tumors are rare soft tissue tumors of submesothelial origin and variable malignant potential. The most common localization is pleural, whereas only 0.6% are of extrapleural localization. Solitary fibrous tumor of the peritoneum, especially of gastric serosa is an extremely rare form of this tumor. Case report. We presented a 65- year-old female patient with solitary fibrous tumor of the stomach. Histopathological analysis of removed tissue showed the presence of tumor tissue built of spindle cells, elongated nuclei with moderately abundant cytoplasm. Cells were in a noncohesive arrangement, in smaller areas distributed in the form of palisade. There were amounts of hipocellular connective tissue, hyalinised, with small foci of dystrophic calcification. Mitoses were rare (less than 3/10 HPF). Blood vessels surrounded the connective tissue. Reviewed material did not contain elements of the parent organ. Immunohistochemically there were positivity on CD34 and vimentin, and negativity to S100, SMA, CD117, dezmin, and Ki-67 is < 2%. The change was diagnosed as a solitary fibrous tumor. Conclusion. Considering that benign solitary fibrous tumors of extrathoracic localizations are extremely rare neoplasms with unpredictable biological behavior and the possibility of recurrence, a long-term clinical and endoscopic follow-up on yearly basis of patients with this disease is recommended. [ABSTRACT FROM AUTHOR]

The immunotoxic effects of tributyltin chloride (TBTC) were examined in the offspring of Sprague–Dawley rats exposed in utero from day 8 of gestation, through lactation and post-weaning until pups reached the age of 30 days (male and female), 60 days (female) and 90 days (male). Daily oral (gavage) doses of 0.025, 0.25 and 2.5 mg/kg body weight/day were administered in olive oil 7 days/week. Immunologic endpoints were investigated at the termination of each study. Statistically significant results (P<0.05) included the following: At 30 days, the mean percent and absolute natural killer (NK) cell numbers were increased in male and female rats treated with the high TBTC dose. At 60 days, female rats had increased mean serum IgM levels at the low and high TBTC doses, increased mean percentage CD4+8+ (immature) T lymphocytes at the middle and high doses, a non-linear dose–response increase in NK cell activity at the 50:1 and 100:1 effector:target cell ratios (pairwise comparisons significant at the low dose compared with control), and increased mean numbers of L. monocytogenes colony-forming bacteria on Day 2 post-infection (significant for trend) and Day 3 post infection (pairwise comparisons significant only in the middle dose). The 90-day male rats had decreased mean serum IgA levels at the middle dose group; increased IgM levels at the high dose group, increased IgG levels at the middle and high doses; decreased IgG2a in the high dose compared to the control; a dose-related increase in the mean percentage NK cell numbers (pairwise comparisons significant at the high dose compared with the control) and increased mean NK cell activity (pairwise comparisons significant at all dose groups compared with the control). The delayed-type hypersensitivity response to oxazolone was increased in the low and middle doses and decreased in the high dose. Thymus atrophy was observed in the high TBTC dose across all ages. Thus, in utero and post-natal treatment of F1 rats with low levels of TBTC affected some aspects of humoral and cell mediated immunity as well as the number and function of cells which are involved in the host''s immunosurveillance mechanisms against tumours and viral infections. [Copyright &y& Elsevier]

Background. Littoral-cell angioma (LCA) is a recently described benign vascular tumor of the spleen, whose imaging and pathologic characteristics have been discussed only by a few authors. The tumor is characterized by a mixture of papillary and cystic areas lined by neoplastic cells deriving from normal splenic lining -- littoral cells. The neoplastic LCA cells express both endothelial and histiocytic antigens associated with CD8 negativity, compared with the normal endothelium of the venous sinuses of the spleen red pulp that only expresses endothelial antigens and CD8 positivity. Therefore, the typical and characteristic immunohistochemical pattern of the LCA is as follows: CD31, CD68, CD163, CD21, FVIII antigen positive; CD34, CD8 negative. Case report. We reported a 60-year-old male with moderate nodular splenomegaly with one large hypoechogenic solid lesion and mild thrombocytopenia in whom the diagnosis of LCA was made after the elective splenectomy. Namely, histopathological and immunohistochemical data allowed a final diagnosis of classical LCA in spite of CD21 negativity. As far as we know this is the first reporeted CD21-negative LCA patient. Histological specimens were presented and differential diagnoses discussed. Conclusion. Littoral-cell angioma is a very rare benign splenic neoplasm that should be considered in the differential diagnosis of multinodular splenomegaly, particularly if the patient has the signs of hypersplenism. [ABSTRACT FROM AUTHOR]

Purpose: In this study, the outcomes and flow cytometry results of our pediatric acute leukemia patients and their relationship with the prognosis and other clinical conditions were investigated. Materials and Methods: A total of 93 patients, 69 acute lymphoblastic leukemia (ALL) and 24 acute myeloid leukemia (AML) diagnosed at our clinic between January 2008 and November 2013, were included. Results: 5-year overall survival (OS) was 66% in ALL patients and 80% in AML patients. When leukemia patients were classified flow cytometrically according to their cell surface antigens (CD); 5-year OS was 58% in CD2(+) patients and 77% in CD2(-) patients. Five-year OS was 82% in patients with CD10(+) and 61% in patients with CD10(-). ALL patients were divided into two groups as ALL with positive T cell markers and ALL with positive B cell markers in the foreground and evaluated accordingly, and the OS of ALL patients with positive T cell markers in the foreground was lower than ALL patients with positive B cell markers in the 5 year (59% and 75%, respectively). Conclusion: In the flow cytometric evaluation of our patients with leukemia, patients with CD10(+) had a better 5-year OS and patients with CD2(+) had a lower 5-year OS. The results of patients with positive B cell markers were better in ALL patients, and the results of patients with positive lymphoid markers were better in AML patients. [ABSTRACT FROM AUTHOR]

Monoclonal antibodies have attracted wide attention in therapeutics owing to their high efficacy, low toxicity, and specific targeting. However, antibodies cannot cross the cell membrane barrier. Therefore, their therapeutic potential is limited to surface-exposed antigens or secreted proteins. In the present investigation, we have developed a chimeric virus-like particle (VLP) of pepper vein banding virus (PVBV) and explored the possibility of using it as a delivery vehicle for antibodies against intracellular antigens as well as for future applications in immunodiagnostics. The chimeric PVBV particles were generated by genetically engineering the B domain of Staphylococcus aureus protein A (SpA) at the N-terminus of the PVBV coat protein (CP). The chimeric VLPs purified by sucrose density gradient centrifugation had ~440-fold higher affinity towards IgG antibody when compared to SpA. Interestingly, the unassembled chimeric CP with the B-domain at the N-terminus (BCP) purified by Ni-NTA chromatography was a monomer, and it had ~45-fold higher affinity towards antibodies compared to SpA. Additionally, the chimeric particles were able to bind and deliver antibodies against both intracellular (α-tubulin) and surface-exposed antigens (CD 20). However, the BCP monomer failed to enter mammalian cells. Thus, for the first time, we have demonstrated that the assembled VLPs are essential for internalization. These results demonstrate the potential of the use of chimeric PVBV VLPs in diagnostics and, more importantly, as nanocarriers for intracellular delivery of antibodies. [ABSTRACT FROM AUTHOR]

Objective To investigate the characteristics of bone marrow mesenchymal stem cell-derived exosomes(BMSC-Exos) and to verify its ability to promote osteogenic differentiation through in vitro and in vivo experiments, as well as to explore its mechanism of promoting bone regeneration. Methods ①Primary human bone marrow mesenchymal stem cells(BMSCs) were cultured and their surface antigen and multilineage differentiation potentials were identified;② Culture supernatants of P4-P6 generation cell culture of BMSC were collected and passed. BMSC-Exos was extracted by the kit method; ③The morphological structure of BMSC-Exos was observed by transmission electron microscope; the surface antigen of BMSC-Exos was detected by immunoelectrophoresis; ④ Alizarin red and ALP staining was used to confirm that BMSC-Exos promoted osteogenic differentiation in vitro; ⑤ The ability of BMSC-Exos to promote bone regeneration was verified in vivo through a rat skull defect animal model;⑥The expression of related proteins and genes in osteoblasts after adding BMSC-Exos was detected by immunoelectrophoresis and qRT-PCR. Results ①The isolated and cultured BMSCs were polygonal or spindle-shaped, and the surface antigens CD 90, CD 29 and CD 44 were positive, which was consistent with the characteristics of mesenchymal stem cells; ② BMSC-Exos was of a biconcave round or oval shape with a diameter of about 40-120 nm(81.7±19.9), and the surface antigen was consistent with BMSC; ③ After Alizarin red staining and ALP staining, the Exo group staining intensity was significantly higher than that of the control group, and the staining intensity was positively correlated with the Exo concentration; ④X-ray and histological analysis of the rat skull defect model showed that the repair of the bone defect in the exosome group was better than that of the control group, and there were more new bone formation; ⑤Immunoelectrophoresis showed that after treatment with exosomes, the content of OCN, Runx 2 and β-catenin protein in MSC cells increased, and qRT-PCR showed that the gene expression of Runx2 and β-catenin was up-regulated. Conclusion BMSC-Exos has the ability to promote bone regeneration and is associated with the up-regulation of Wnt/β-catenin pathway. [ABSTRACT FROM AUTHOR]

Purpose: Reliable biomarkers for predicting subsequent sepsis among patients with suspected acute infection are lacking. In patients presenting to emergency departments (EDs) with suspected acute infection, we aimed to evaluate the reliability and discriminant ability of 47 leukocyte biomarkers as predictors of sepsis (Sequential Organ Failure Assessment score ≥ 2 at 24 h and/or 72 h following ED presentation).Methods: In a multi-centre cohort study in four EDs and intensive care units (ICUs), we standardised flow-cytometric leukocyte biomarker measurement and compared patients with suspected acute infection (cohort-1) with two comparator cohorts: ICU patients with established sepsis (cohort-2), and ED patients without infection or systemic inflammation but requiring hospitalization (cohort-3).Results: Between January 2014 and February 2016, we recruited 272, 59 and 75 patients to cohorts 1, 2, and 3, respectively. Of 47 leukocyte biomarkers, 14 were non-reliable, and 17 did not discriminate between the three cohorts. Discriminant analyses for predicting sepsis within cohort-1 were undertaken for eight neutrophil (cluster of differentiation antigens (CD) CD15; CD24; CD35; CD64; CD312; CD11b; CD274; CD279), seven monocyte (CD35; CD64; CD312; CD11b; HLA-DR; CD274; CD279) and a CD8 T-lymphocyte biomarker (CD279). Individually, only higher neutrophil CD279 [OR 1.78 (95% CI 1.23-2.57); P = 0.002], higher monocyte CD279 [1.32 (1.03-1.70); P = 0.03], and lower monocyte HLA-DR [0.73 (0.55-0.97); P = 0.03] expression were associated with subsequent sepsis. With logistic regression the optimum biomarker combination was increased neutrophil CD24 and neutrophil CD279, and reduced monocyte HLA-DR expression, but no combination had clinically relevant predictive validity.Conclusions: From a large panel of leukocyte biomarkers, immunosuppression biomarkers were associated with subsequent sepsis in ED patients with suspected acute infection.Clinical Trial Registration: NCT02188992. [ABSTRACT FROM AUTHOR]

Flow cytometry is a ubiquitous, multiparametric method for characterizing cellular populations. However, this method can grow increasingly complex with the number of proteins that need to be screened simultaneously: spectral emission overlap of fluorophores and the subsequent need for compensation, lengthy sample preparation, and multiple control tests that need to be performed separately must all be considered. These factors lead to increased costs, and consequently, flow cytometry is performed in core facilities with a dedicated technician operating the instrument. Here, we describe a low-cost, label-free microfluidic method that can determine the phenotypic profiles of single cells. Our method employs Node-Pore Sensing to measure the transit times of cells as they interact with a series of different antibodies, each corresponding to a specific cell-surface antigen, that have been functionalized in a single microfluidic channel We demonstrate the capabilities of our method not only by screening two acute promyelocytic leukemia human cells lines (NB4 and AP-1060) for myeloid antigens, CD 13, CD14, CD15, and CD33, simultaneously, but also by distinguishing a mixture of cells of similar size--AP-1060 and NALM-1 based on surface markers CD 13 and HLA-DR. Furthermore, we show that our method can screen complex subpopulations in clinical samples: we successfully identified the blast population in primary human bone marrow samples from patients with acute myeloid leukemia and screened these cells for CD 13, CD34, and HLA-DR. We show that our label-free method is an affordable, highly sensitive, and user-friendly technology that has the potential to transform cellular screening at the benchside. [ABSTRACT FROM AUTHOR]

Purpose: Considering the important role of neutrophils' activation in the pathogenesis of acute pancreatitis (AP), the aim of our study was to evaluate the expression of leukocytes' adhesion molecules in patients with AP. Patients/methods: Thirty-five patients (16 women and 19 men; age 32-77 years, median 56 years) with AP were prospectively included into our study. The absolute number of leukocytes was estimated by haematologic analyser. Surface neutrophils antigens (CD) were assayed by the direct fluorescence method for whole blood, using a flow cytometer. Results: At the day 1, significant increase of ICAM-1 expression was found in patients with severe AP (S-AP) (7280 mm-3 vs 2850 mm-3 in healthy control; p < 0.05). In the days 2, 3 and 5 it sharply decreased and peaked again to 4860 mm-3 at the day 10. In patients with mild AP (M-AP), not significant elevation of ICAM-1 quickly returned to normal level. In both forms of AP, neutrophil CD62L (L-selectin) expression reached the highest level at the day 1 (8800 mm-3 and 9020 mm-3, respectively in M-AP and S-AP, in comparison to 3400 mm-3 in control; p < 0.05). Expression of CD69 (neutrophils' marker of early activation) significantly increased in both M-AP and S-AP. Conclusions: We have found an early and significant increase of peripheral blood neutrophil CD54/ICAM- 1 expression, specific for S-AP but not for M-AP. It may provide a good marker predicting severe course of pancreatitis. [ABSTRACT FROM AUTHOR]

Background aims. Adipose tissue is a rich and very convenient source of cells for regenerative medicine therapeutic approaches. However, a characterization of the population of adipose-derived stromal and stem cells (ASCs) with the greatest therapeutic potential remains unclear. Under the authority of International Federation of Adipose Therapeutics and International Society for Cellular Therapy, this paper sets out to establish minimal definitions of stromal cells both as uncultured stromal vascular fraction (SVF) and as an adherent stromal/stem cells population. Methods. Phenotypic and functional criteria for the identification of adipose-derived cells were drawn from the literature. Results. In the SVF, cells are identified phenotypically by the following markers: CD45-CD235a-CD31-CD34+. Added value may be provided by both a viability marker and the following surface antigens: CD 13, CD73, CD90 and CD 105. The fibroblastoid colony-forming unit assay permits the evaluation of progenitor frequency in the SVF population. In culture, ASCs retain markers in common with other mesenchymal stromal/stem cells (MSCs), including CD90, CD73, CD 105, and CD44 and remain negative for CD45 and CD31. They can be distinguished from bone-marrow-derived MSCs by their positivity for CD36 and negativity for CD 106. The CFU-F assay is recommended to calculate population doublings capacity of ASCs. The adipocytic, chondroblastic and osteoblastic differentiation assays serve to complete the cell identification and potency assessment in conjunction with a quantitative evaluation of the differentiation either biochemically or by reverse transcription polymerase chain reaction. Conclusions. The goal of this paper is to provide initial guidance for the scientific community working with adipose-derived cells and to facilitate development of international standards based on reproducible parameters. [ABSTRACT FROM AUTHOR]

Abstract: This study was conducted to assess the effect of modifying the n-6:n-3 polyunsaturated fatty acid ratio (FAR) of diet using linseed, soybean, and corn oil on growth performance, cardiac antioxidant status, T-cell and cytokine mRNA expression in the thymus, and blood T lymphocyte subset characteristics in broilers. A total of 480 one-day-old male broilers (BW=46±4g) were randomly allotted into 4 treatments consisting of dietary n-6 to n-3 FAR of 12:1, 9:1, 6:1, and 3:1. There were 6 replicates per treatment with 20 birds per replicate. After feeding for 42d, chicks were slaughtered, and the blood and tissue samples were collected. Decreasing n-6 to n-3 FAR did not affect feed intake, but increased BW gain (linear, P=0.05) and improved feed conversion ratio (quadratic, P=0.03) during the first 3 weeks. Cardiac hydroxyl radical (P=0.03) and malondialdehyde (MDA) (P=0.01) concentration reduced linearly with decreasing dietary n-6:n-3 FAR, whereas superoxide dismutase (SOD) (quadratic, P=0.04), glutathione S-transferase (GSH-ST) (quadratic, P=0.02) and glutathione peroxidase (GSH-Px) (linear, P=0.05) levels were increased. The T lymphocyte subsets were also analyzed using monoclonal antibodies and flow cytometry simultaneously. The messenger RNA (mRNA) expression of cluster of differentiation antigens (CD)4+ receptor and toll-like receptor (TLR)-3 in thymus were depressed (quadratic, P<0.05) by decreasing n-6:n-3 FAR, whereas the expression of interferon (IFN)-α was unaffected by dietary treatment. Concentration of blood CD4+, CD8+ and CD4+:CD8+ ratio linearly declined (P<0.05) as the n-6:n-3 FAR decreased. In conclusion, the decreasing n-6:n-3 FAR can enhance the levels of cardiac antioxidant enzymes, and affect thymus immune function of fast growing broilers. [Copyright &y& Elsevier]

The objective of this study was to analyze the relationships among immunohistochemical expression of p53, Ki-67, Fas antigens (CD 95) and tumor size, pathological stage, and the presence of vascular invasion and necrosis in testicular germ-cell tumors. Representative archival tissues from 36 testicular germ cell tumors were studied for p53, Ki-67 and Fas antigens by immunohistochemistry. All of the tumors' size, pathological stage, and the presence of vascular invasion and necrosis within tumors were recorded. Significant correlation with pathological phase was found for Ki-67 and Fas expressions. No correlation was determined between any antibody and maximum tumor diameter or necrosis. While a positive correlation (p<0.05) with vascular invasion in mixed germ-cell tumors was found with Ki-67 staining intensity, no significant correlation (p>0.05) was found with Fas and p53. In the previous and current studies, the percentage of embryonal carcinoma and the presence or absence of vascular invasion in NSGCT appear to be reliable prognostic factors to identify patients at high risk and low risk for occult retroperitoneal disease. Additionally, our results, suggest that Ki-67 plays a role at all stages of tumorogenesis and that it could be used as an auxiliary to other parameters in tumor prognosis. It was concluded that Ki-67 and Fas expressions may be important prognostic parameters in the staging of germ-cell tumors. [ABSTRACT FROM AUTHOR]

The leukocyte adhesion 90-kDa glycoprotein GP90 (antigen CD18) is non-covalently associated separately with cell-surface glycoproteins GP160 (antigens CD11a, TA-1, LFA-1), GP155 (antigens CD11b, OKM1, MO1) or GP130 (antigens CD 11c, Leu-M5). Large amounts of these protein complexes were purified to homogeneity from blood mononuclear leukocytes by immunoaffinity chromatography using a monoclonal antibody. GP90 was further isolated by preparative gel electrophoresis in the presence of sodium dodecyl sulfate. Rabbit antiserum towards the complex inhibited phorbol-ester-induced adhesion of leukocytes. The antiserum towards purified GP90 reacted more strongly with the denatured GP90 protein, but showed reactivity also with GP160 protein, indicating structural homologies between GP90 and GP160. The amino acid composition of GP90 was determined. Its N-terminus was found to be blocked. Treatment of GP90 with endo-β-N-acetylglucosaminidase F, but not with endo-β-N-acetylglucosaminidase H, reduced the apparent molecular mass to 75 kDa, indicating the presence of five or six N-linked complex-type oligosaccharides/molecule. [ABSTRACT FROM AUTHOR]

Abstract: Inflammation protects from dangerous stimuli, restoring normal tissue homeostasis. Inflammatory response in the nervous system (“neuroinflammation”) has distinct features, which are shared in several diseases. The retina is an immune-privileged site, and the tight balance of immune reaction can be disrupted and lead to age-related macular disease (AMD) and to its peculiar sign, the druse. Excessive activation of inflammatory and immunological cascade with subsequent induction of damage, persistent activation of resident immune cells, accumulation of byproducts that exceeds the normal capacity of clearance giving origin to a chronic local inflammation, alterations in the activation of the complement system, infiltration of macrophages, T-lymphocytes and mast-cells from the bloodstream, participate in the mechanisms which originate the drusen. In addition, aging of the retina and AMD involve also para-inflammation, by which immune cells react to persistent stressful stimuli generating low-grade inflammation, aimed at restoring function and maintaining tissue homeostasis by varying the set point in relation to the new altered conditions. This mechanism is also seen in the normal aging retina, but, in the presence of noxious stimuli as in AMD, it can become chronic and have an adverse outcome. Finally, autophagy may provide new insights to understand AMD pathology, due to its contribution in the removal of defective proteins. Therefore, the AMD retina can represent a valuable model to study neuroinflammation, its mechanisms and therapy in a restricted and controllable environment. Targeting these pathways could represent a new way to treat and prevent both exudative and dry forms of AMD. [Copyright &y& Elsevier]