Epstein-Barr virus-encoded miR-BART11 promotes tumor-associated macrophage-induced epithelial-mesenchymal transition via targeting FOXP1 in gastric cancer.

Gastric carcinoma (GC) is an Epstein-Barr virus (EBV)-associated malignancy characterized by early metastasis. Unlike that of cellular micro(mi)RNAs, the role of viral miRNAs in epithelial-mesenchymal transition (EMT) and metastasis in cancers has not been fully investigated. In this study, we elucidated the involvement of miR-BART11, an EBV-encoded viral miRNA, in the EMT and metastasis of GC cells. EBV-miR-BART11 upregulation can lead to downregulation of forkhead box protein P1 (FOXP1) in both tissues and cell lines of gastric carcinoma. Downregulation of FOXP1 might trigger the secretion of interleukin 1ß (IL-1ß), IL-6, and 1L-10 in cancer cells, resulting in poor survival of GC patients. We found that the observed EMT phenotypes resulted from the EBV-miR-BART11 overexpression-induced FOXP1 downregulation, which impacted the expression of the EMT-transcription factors E-cadherin and snail. We further demonstrated that conditioned medium-derived tumor-associated macrophages (TAMs) promoted phenotypic changes and expression of EMT-related molecules in GC cells. Additionally, EMT changes were significantly promoted in GC cells cultured in conditioned medium from TAMs infected with EBV-miR-BART11-containing lentivirus. On the contrary, GC cells cultured in conditioned medium from TAMs infected with FOXP1-carrying lentivirus showed little or no EMT change. Taken together, our results suggest that EBV-encoded viral miRNA BART11 downregulates the FOXP1 transcription factor, and promotes EMT by directly influencing gastric tumor cells or indirectly affecting the tumor microenvironment, which might, in turn, accelerate cancer invasion and metastasis, thereby affecting the survival and prognosis of patients.

The combination of Caprini risk assessment scale and thrombotic biomarkers to evaluate the risk of venous thromboembolism in critically ill patients.

To evaluate the correlation between the Caprini risk assessment scale and plasma thrombosis biomarkers and estimate the validity of this method in identifying critically ill patients at high risk of venous thromboembolism (VTE).Patients with VTE who were admitted to the intensive care unit (ICU) department of West China Hospital SiChuan University from October 2016 to October 2017 were enrolled in this case-control study. We retrieved relative clinical data and laboratory test results included in the Caprini risk assessment scale to calculate the Caprini score and compared thrombosis biomarkers between various risk stratifications (low, moderate, high, and highest).A total of 151 critically ill patients were enrolled in our research, including 47 VTE and 94 non-VTE patients. The differences in Caprini score and levels of thrombosis biomarkers between the VTE and control group were significant. Thrombomodulin (TM) was positively correlated with Caprini score (R-value was .451, P < .05). Based on the receiver operating characteristic analysis, TM, tissue plasminogen activator-inhibitor complexes, D-dimer, and fibrinogen degradation products had a certain diagnostic efficiency in distinguishing VTE from others (P < .05). Using the logistic regression model, we identified that 5 risk factors, namely drinking history, major surgery (>3 hours), swollen legs (current), TM, and D-dimer, were independent factors for the occurrence of VTE in critically ill patients admitted in the ICU.Thrombosis markers were positively correlated with Caprini risk stratification. The combination of plasma markers and Caprini risk assessment scale can further increase the predictive value in critically ill patients with VTE.

[Value of Coagulation and Fibrinolysis Biomarker in Lung Cancer Patients with Thromboembolism].

BACKGROUND: Coagulation and fibrinolysis biomarkers can effectively reflect the dysfunction of coagulation and anticoagulation system, and the changes of their levels were closely related to the hypercoagulable status. The aim of this study is to study the variation tendency of these coagulation and fibrinolysis markers and explore the diagnosis power and clinical value of these biomarker for thrombosis in postoperative lung cancer patients with deep vein catheterization. METHODS: We selected 118 postoperative lung cancer patients with deep vein catheterization including 29 patients with thromboembolism and 89 patients in control group. Coagulation and fibrinolysis parameters [thrombomodulin (TM)/thrombin-antithrombin complex (TAT)/α2-plasmin inhibitor-plasmin complexes (PIC)/tissue plasminogen activator-inhibitor complexes (t-PAIC)] and traditional coagulation time[prothrombintime (PT)/activated partial thrombo plastin time(APTT)/thrombintime (TT)/fibrinogen (FIB)/antithrombin III (ATIII)/fibrinogen degradation products (FDP)/D-Dimer (D-D)] were detected in both groups. We analyzed the variation tendency of these biomarkers and figured out the diagnosis powerfor thrombosis. RESULTS: A statistically significant difference was available on the value of TM, TAT, PIC, t-PAIC, D-D, FDP between thrombosis group and non-thrombosis group (P<0.05). TM, TAT, PIC, t-PAIC, D-D, FDP performed with an AUC of 0.770, 0.771, 0.669, 0.671, 0.819, 0.816, respectively (P<0.05). CONCLUSIONS: An enhanced coagulation and fibrinolysis activity existed in lung cancer patients with deep vein catheterization after surgery, and early detection of coagulation and fibrinolytic biomarkers could prevent thrombosis and reduce postoperative thrombosis complications in patients with lung cancer.

Enhanced IL-6/phosphorylated STAT3 signaling is related to the imbalance of circulating T follicular helper/T follicular regulatory cells in patients with rheumatoid arthritis.

BACKGROUND: Follicular helper T (Tfh) cells are specialized in helping B lymphocytes, which play a central role in autoimmune diseases that have a major B cell component, such as in rheumatoid arthritis (RA). Follicular regulatory T (Tfr) cells control the over-activation of Tfh and B cells in germinal centers. Dysregulation of Tfh cells and Tfr cells has been reported to be involved in the pathogenesis of some autoimmune diseases. However, the balance of Tfh and Tfr cells, and their roles in the development and progression of RA are still not clear. METHODS: In this study, we enrolled 44 patients with RA (20 patients with active RA and 24 patients with inactive RA) and 20 healthy controls, and analyzed the frequencies of circulating Tfh and Tfr cells, expression of programmed death-1 (PD-1), inducible co-stimulator (ICOS), intracellular IL-21, and pSTAT3 in Tfh cells, and serum levels of IL-6. The correlation among these parameters and that of Tfh or Tfr cells with disease activity were also analyzed. RESULTS: Patients with RA (especially active RA) had higher frequencies of Tfh cells, but lower percentages of Tfr cells, thereby resulting in elevated ratios of Tfh/Tfr. Expression levels of PD-1 and IL-21 in Tfh cells were higher in patients with RA than in healthy subjects, while no difference in ICOS expression was observed between patients and controls. Both pSTAT3 expression and serum IL-6 levels increased in patients with RA, and positive correlation between them was observed. Additionally, pSTAT3 expression was positively correlated with Tfh cell frequency. The Disease Activity Score in 28 joints based on C-reactive protein (DAS28-CRP) was negatively correlated with Tfr cell frequency, but was positively correlated with both Tfh/Tfr ratio and PD-1 expression. CONCLUSIONS: Results demonstrated that enhanced IL-6/pSTAT3 signaling may contribute to promotion of Tfh cells, consequently skewing the ratio of Tfh to Tfr cells, which may be crucial for disease progression in RA.

[Adjusting Platelet Counts for Platelet Aggregation Tests].

OBJECTIVE: To explore a better method to adjust platelet counts for light transmission aggregometry (LTA). METHODS: Blood samples from 36 healthy participants aged from 18 to 50 yr. were collected.Platelet-rich plasma (PRP) was diluted using platelet-poor plasma (PPP) and physiological saline (PS),respectively,in a ratio of 1.5,2,2.5 and 3 times. Platelet aggregation was induced by adenosine diphosphate (ADP),arachidonic acid (ARA),collagen (COL), epinephrine (EPI),or ristocetin (RIS). The maximal aggregation rates (MAs) of different approaches were compared. We also compared the MAs induced by RIS between PRP-obtained-PPP and whole blood-obtained-PPP (2 100×g, 5 min). RESULTS: Compared with the original PRP,the MAs induced by ADP,ARA,and EPI decreased in PPP-adjusted PRP (significant at 2-3 times dilution ratio,P<0.05),but not in PS-adjusted PRP (P>0.05). The MA induced by RIS decreased in PS-adjusted PRP (significant at all dilution ratios,P<0.05),but not in PPP-adjusted PRP (P>0.05). No changes in the MA induced by COL were found in PS-adjusted PRP and PPP-adjusted PRP (P>0.05). Whole blood-obtained-PPP (2 100×g, 5 min) had the same MA induced by ristocetin compared with PRP-obtained-PPP (P>0.05). CONCLUSION: PS is recommended for adjusting platelets counts for platelet aggregation induced by ADP,ARA,COL and EPI. Whole blood-obtained-PPP (2 100 ×g, 5 min) is recommended for RIS-induced aggregation as a matter of convenience.

[Clinical Significance of Coagulation Indicators and their Correlation with Inflammatory Factors in Critical Patients with Thromboembolism].

OBJECTIVE: To evaluate the levels of coagulation indicators [thrombomodulin(TM)/ thrombin-antithrombin complexes(TAT)/ α2-plasmin inhibitor-plasmin complexes(PIC)/ tissue plasminogen activator-inhibitor complexes(t-PAIC) /D-Dimer(D-D)/fibrin degradation products(FDP)] in the critical patients with thromboembolism, analyse their correlation with inflammatory factor (procalcitonin/C reactive protein/ interleukin-6), and explore the diagnostic significance of coagulation indicators for these patients. METHODS: The serum levels of the coagulation indicators (TM/TAT/PIC/t-PAIC/D-D /FDP) and inflammatory factors (PCT/IL-6/CRP) were detected in the patient group with critical thromboembolism (n= 38) and critical patient group without thromboembolism as control (n= 81) . The correlation of coagulation indicators with inflammatory factors was analyzed. RESULTS: The values of TM/TAT/PIC/D-D/FDP in thromboembolism group were statistically significantly higher than those in control group (P<0.05). However, the t-PAIC values were not significantly different (P>0.05), and 3 inflammatory factors (PCT/CRP/IL-6) in thromboembolism patients were significantly higher than those in control group (P<0.05). The correlation analysis suggested that the correlation coefficients of TM with PCT, CRP and IL-6 were 0.288, 0.249 and 0.270, respectively (P<0.05). CONCLUSION: The critical patients with thromboembolism show an obviously higher systemic inflammatory response, and accompany with coagulation dysfunction. There is a network relationship between inflammation and coagulation, the interaction of inflammatory factors with coagulation indicators promotes thromboembolism and inflammation.

Capillarys 2 Flex Piercing Detected a Rare Case of Hb Broomhill.

We report a case of an extremely rare hemoglobin (Hb) variant-Hb Broomhill, which has been only reported once in the literature. Hemoglobin fractions were determined by capillary electrophoresis (Sebia Capillarys 2 Flex piercing) and high performance liquid chromatography (HPLC) (Bio-Rad Variant™ II Hemoglobin Testing System), respectively. Complete blood count and DNA sequencing were also performed. The capillary electrophoregram revealed a tiny shoulder peak before the HbA peak and a subtle abnormal HbA2 peak (slightly wider and lower), even though the percentage of each hemoglobin fraction was within the reference range (HbA, 97.4%; HbA2, 2.6%). On HPLC, not only the percentage but also the peak shape of each hemoglobin fraction was normal (HbA 88.2%, HbA2 2.5%, HbF 0.6%). Eventually, sequencing analysis of α genes confirmed a missense mutation (CCC>GCC at codon 114 in alpha1 gene) which caused Hb Broomhill variant. Our report suggest that capillary electrophoresis may be an accurate tool for screening and diagnosis of Hb disorders.