[The clinical significance of CD(4)(+)CD(69)(+) T lymphocytes in patients with autoimmune hemolytic anemia/Evans syndrome].

Objective: To investigate the significant of peripheral CD(4)(+) CD(69)(+) T lymphocytes in patients with autoimmune hemolytic anemia (AIHA)/Evans syndrome (ES). Methods: In this study peripheral blood samples from 32 patients with AIHA/ES (15 hemolytic episode patients, 17 remission patients) and 13 healthy controls were collected. Patients with AIHA/ES were recruited in Tianjin Medical University General Hospital from October 2015 to May 2016. The percentages of CD(69)(+) T lymphocytes were analyzed by flow cytometry. The expression of CD(69) mRNA in CD(4)(+) T lymphocytes which was sorted from peripheral blood by magnetic activated cell sorting (MACS) was detected using real-time PCR. Soluable CD(69) was measured by ELISA. Results: In hemolytic episode patients, the ratio of CD(3)(+)CD(69)(+)/CD(3)(+)T lymphocytes [(3.08±1.48)%] was significantly higher than that in healthy controls [(1.28±0.83)%, P<0.01] and in remission group[(1.96±1.33)%, P<0.05]. The absolute count of CD(3)(+)CD(69)(+)T lymphocytes in hemolytic episode group [(2.94±1.81)×10(7)/L] was higher than that in healthy controls [(1.48±1.42)×10(7)/L, P<0.05]. The ratio of CD(3)(+)CD(4)(+)CD(69)(+)/CD(3)(+)CD(4)(+)T cells in hemolytic episode group [(2.16±1.56)%] was significantly higher than that in remission group [(1.16±0.62)%, P<0.05] and healthy controls[(0.94±0.78)%, P<0.05]. The quantity of CD(3)(+)CD(4)(+)CD(69)(+)T lymphocytes in hemolytic episode group[(1.04±0.98)×10(7)/L] was higher than in healthy controls [(0.44±0.38)×10(7)/L, P<0.05]. The ratio of CD(3)(+)CD(8)(+)CD(69)(+)/CD(3)(+)CD(8)(+)T lymphocyte in hemolytic episode group [(4.87±2.56)%] was significantly higher than that in healthy controls[(1.83±1.27)%, P<0.01]. The quantity of CD(3)(+)CD(8)(+)CD(69)(+)T lymphocytes in three groups did not show significant difference. The ratio of CD(3)(+)CD(4)(+)CD(69)(+)/CD(3)(+)CD(4)(+) T lymphocytes in hemolytic episode group was negatively correlated with hemoglobin (Hb) (P<0.01) , positively correlated with the percentage of reticulocytes (Ret%) (P=0.01) total bilirubin(TBil), indirect bilirubin(IBil) (P<0.01) and not correlated with absolute reticulocytes count, lactic dehydrogenase (LDH), complement 3(C3), complement 4 (C4). The ratio of CD(3)(+)CD(4)(+)CD(69)(+)/CD(3)(+)CD(4)(+)T lymphocytes in remission group was negatively correlated with Hb (P<0.05). In hemolytic episode patients CD(69) mRNA (32.26±35.11) was significantly higher than that in remission group(6.05±5.87) (P<0.05) and healthy controls (1.76±1.85)(P<0.01). CD(69) mRNA in remission group was significantly higher than healthy controls (P<0.05). Serum CD(69) in hemolytic episode patients [(494.21±16.06) ng/L] was significantly higher than that in healthy controls [(441.39±104.6) ng/L, P<0.05]. Conclusion: Our findings suggest that the proportion of CD(4)(+)CD(69)(+) T lymphocytes increase in AIHA/ES patients, which is correlated with the severity of disease.

[Which animal models of nonalcoholic steatohepatitis-hepatocellular carcinoma (NASH-HCC) have more practical value in experimental investigations?].

In the past, we used to think that hepatocellular carcinoma (HCC) only occurred when nonalcoholic fatty liver disease (NAFLD) progressed to liver cirrhosis. However, recent clinical studies have shown that HCC may occur during the stage of nonalcoholic steatohepatitis (NASH). The prevalence of NAFLD and the increasing incidence of NASH-associated HCC require the elucidation of the pathogenesis of NASH-HCC and the assessment of the efficacy of novel therapies as early as possible, while these evaluations need reliable animal models. This article reviews the characteristics of NASH-HCC models with a practical value to uncover the mysteries of NASH-HCC.

Aberrant hedgehog signaling is responsible for the highly invasive behavior of a subpopulation of hepatoma cells.

Hepatoma exhibits a series of heterogeneous subpopulations in its cell surface markers, tumorigenicity, invasion and metastatic capability. We previously demonstrated that the CD133(-)/EpCAM(-) hepatoma subpopulation was more metastatic than its counterpart; however, the controlling mechanisms are unexplored. The present study aimed to delineate the significance of aberrant hedgehog (Hh) signaling in the mediation of metastases. Fluorescence-activated cell sorting-enriched CD133(-)/EpCAM(-) (double negative, DN), Huh-7 cells underwent a transwell selection for metastatic cells (transwell-selected, TS). The TS cells displayed much greater metastatic activity as evidenced by an increased invasion rate, extremely upregulated expression of matrix metalloproteinase (MMP)-1/2/9 genes compared with DN and double-positive (DP) subpopulations. In contrast to DP cells, TS cells lost E-cadherin and were all vimentin-positive as shown by immunocytochemistry. There was a transitional increase in Gli-1/2 gene expression levels from DP, DN to TS subpopulations, which was consistent with elevated Gli-1/2 or Twist-1 protein levels in the nuclear fraction. Furthermore, truncated Gli-1 (tGli-1), which transactivates molecules involved in metastasis, was detected in the highly invasive Huh-7 cell subpopulation, but not in less metastatic hepatoma cells or normal hepatocytes. The enhanced metastatic features with increased expression of MMPs as well as the presence of twist and snail genes in TS Huh-7 cells were reversed by LDE225, a potent Smoothened antagonist. In conclusion, the highly metastatic capability of a unique TS subpopulation was highly attributed to significant epithelial-mesenchymal transition, enhanced Hh activity and aberrant occurrence of a tGli-1 variant, which appears to be responsible for the highly invasive behavior.