Transfer of CD11c+ lamina propria mononuclear phagocytes from post-infectious irritable bowel syndrome causes mucosal barrier dysfunction and visceral hypersensitivity in recipient mice.

The role of low-grade inflammation in the development of post­infectious irritable bowel syndrome (PI­IBS) has attracted increasing attention. Abnormal CD11c+ mononuclear phagocytes, such as dendritic cells (DCs), macrophages, and monocytes, are involved in the disruption of immune tolerance in organisms, which can lead to the development of chronic inflammatory diseases. The present study tested the hypothesis that CD11c+ lamina propria mononuclear phagocytes (CD11c+ LPMPs) contribute to increased mucosal permeability and visceral hypersensitivity in a PI­IBS mouse model. CD11c+ LPMPs were isolated and purified via the digestion of intestinal tissues and magnetic­activated cell sorting. We detected increased mucosal permeability, visceral hypersensitivity and intestinal inflammation during both the acute and chronic stages of Trichinella infection. Following the transfer of CD11c+ LPMPs from PI­IBS mice into normal mice, low­grade inflammation was detected, as demonstrated by increased IL­4 expression in the ileum, as well as enhanced mucosal permeability, as indicated by decreased transepithelial electrical resistance and the pre-sence of ultrastructural alterations. More importantly, the mice that underwent adoptive transfer of CD11c+ LPMPs from the PI­IBS mice also exhibited increased abdominal withdrawal reflex scores and a decreased threshold. Our data demonstrated that the CD11c+ LPMPs from this PI­IBS mouse model were not only able to transfer enteric inflammation to the normal mice but also caused abnormal intestinal function, characterized by epithelial barrier disruption and visceral hyperalgesia.

Expression of ß-catenin protein in hepatocellular carcinoma and its relationship with alpha-fetoprotein.

This study aimed to investigate the expression of ß-catenin in hepatocellular carcinoma (HCC) tissues and its relationship with α-fetoprotein (AFP) in HCC. Immunohistochemistry was used to determine the expression of ß-catenin in normal liver tissues (n=10), liver cirrhosis tissues (n=20), and primary HCC tissues (n=60). The relationship between ß-catenin expression and clinical parameters of HCC was investigated. Real-time PCR and Western blotting were used to detect the mRNA and protein expression levels of ß-catenin in the liver cancer cell line SMMC-7721 transfected with a plasmid encoding AFP, and also the mRNA and protein expression levels of ß-catenin were measured in the liver cancer cell line Huh7 before and after the transfection with AFP shRNA plasmids. The results showed that ß-catenin was only expressed on the cell membrane in normal liver tissues. Its localization to the cytoplasm and nucleus of cells was observed in a small proportion of cirrhotic tissues or adjacent HCC tissues, and such ectopic expression of ß-catenin was predominant in HCC tissues. The abnormal expression of ß-catenin was correlated with serum AFP levels, cancer cell differentiation and vascular invasion (P<0.05). Additionally, the increased expression of AFP resulted in the upregulation of ß-catenin mRNA and protein levels, while knockdown of AFP with AFP shRNA led to significantly decreased ß-catenin mRNA and protein levels (P<0.05). It was suggested that the abnormal expression of ß-catenin is implicated in hepatic carcinogenesis and development. AFP can lead to increased expression of ß-catenin, which may account for the poor prognosis of AFP-associated HCC patients.

Overexpression of eIF4E gene in acute myeloid leukemia and its relation with disease progression / 中国实验血液学杂志

The aim of this study was to detect the expression level of eIF4E gene in patients with non-treated, remission and non-remission/relapse acute myeloid leukemia (AML), and other non-malignant haematologic diseases so as to analyze and reveal the relationship of eIF4E gene expression with AML progression. SYBR Green I RT-PCR was used to assay the expression level of eIF4E mRNA extracted from bone marrow mononuclear cells in 30 patients with AML (6 in M2, 5 in M3, 8 in M4, 10 in M5, 1 in M6) and 20 patients with non-malignant hematologic diseases. The β2-microglubin(β2M) was used as internal reference and the formula 2(-ΔCt)×100% was applied to calculate the expression level of eIF4E gene. The results showed that the eIF4E expression level (7.098 ± 5.544)% in patients with non-treated and non-remitted/relapsed AML was significantly higher than that in patients with remission (0.964 ± 0.312)% (P < 0.01) and non-malignant hematologic diseases (0.248 ± 0.163)% (P < 0.01). There was no difference between latter two group patients, even though the expression level of eIF4E gene in patients with M4 and M5 was higher. As compared with non-malignant hematologic diseases, the expression level of eIF4E gene of patients with remission patients showed no significant difference. It is concluded that the over-expression of eIF4E gene has been found in patients with AML, and its level obviously decreases along with remission of disease, thus the eIF4E gene may be a surveillance parameter for disease progression.

Change of expression pattern of CD3 genes in peripheral blood T-cells from CML patients / 中国实验血液学杂志

Our previous finding showed that down-regulation of CD3ζ gene was detected in patients with chronic myeloid leukemia (CML). In order to further elucidate the feature of T cell immune status in the signal transduction in CML patients, the expression patterns of all 4 CD3 genes were characterized in peripheral blood of patients, the expression levels of CD3γ, δ, ε and ζ chain genes were detected by real time qPCR with SYBR Green I staining in peripheral blood mononuclear cells (PBMNCs) from 17 cases of de novo CML patients in chronic phase and 17 cases of healthy individuals, the ß₂-microglobulin gene was used as an internal reference, and the mRNA expression level of each CD3 gene was evaluated by the 2(-ΔCt) x 100% method. The results showed that the median expression levels of CD3γ, δ and ε genes (2.344%, 0.515% and 3.516%) in CML patients were not significantly different from healthy individuals (p = 0.072, p = 0.190, p = 0.615, respectively), while the expression level of CD3ζ gene in PBMNCs from CML patients (0.395%) was lower than that from healthy individuals (1.538%) (p < 0.001). The expression patterns of 4 CD3 genes in proper order were CD3ε > CD3γ > CD3δ > CD3ζ in CML group, in contrast, the expression patterns were presented as CD3γ > CD3ε > CD3ζ > CD3δ in healthy group. It is concluded that the present study characterized the expression pattern of CD3γ, δ, ε and ζ chain genes in CML patients, lower expression of CD3ζ is the feature of TCR signal transduction immunodeficiency and the expression patterns of 4 CD3 genes are changed in CML patients.

Mechanism of multidrug resistance of human small cell lung cancer cell line H446/VP / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Small cell lung cancer (SCLC) is the most aggressive form of lung cancer. This study aimed to investigate the mechanism of human small cell lung cancer cell line resistance to etoposide (VP-16), H446/VP.</p><p><b>METHODS</b>The cell viability was measured by MTT assay. Immunocytochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting methods were used to detect the multidrug resistance gene (MDR1), bcl-2, bax and the topoisomerase II (Topo II) expressions in H446 and H446/VP cells after treated with or without VP-16.</p><p><b>RESULTS</b>The 50% inhibition concentration (IC50) of VP-16 on H446 cells was 49 mg/L, and 836 mg/L was for H446/VP cells. The expressions of MDR1 and bcl-2 were up-regulated, while the amounts of bax and Topo II were reduced in H446/VP cells. After treated with 49 mg/L of VP-16, it showed that the drug could significantly inhibit bcl-2 and Topo II expressions, and increase bax expression in H446 cells compared with that of H446/VP cells.</p><p><b>CONCLUSIONS</b>The H446/VP cell was stably resistant to VP-16. The decreased expression of Topo II was correlated with the H446/VP multidrug resistance. The elevated expressions of MDR1, and the altered apoptotic pathways also played an important role in VP-16 induced multidrug resistance of SCLC.</p>

Analysis of selected usage and clonal expansion of TCR Vbeta repertoire of peripheral blood T cells in patients with non-Hodgkin's lymphoma / 中国实验血液学杂志

To investigate the distribution and clonal expansion of TCR Vbeta subfamily T cells in patients with B-NHL and T-NHL, the CDR3 of TCR Vbeta 24 subfamily genes was amplified in peripheral blood mononuclear cells from 4 cases with B-NHL and 2 cases with T-NHL using RT-PCR, and to observe the usage of TCR Vbeta repertoire, the PCR products were further labeled with fluorescence and analyzed by genescan technique for the CDR3 size, to evaluating clonality of the detectable TCR Vbeta T cells. The results indicated that only selected expression of 6-12 Vbeta subfamily T cells could be identified in the 6 cases with NHL, and Vbeta1, Vbeta8, Vbeta13 and Vbeta19 were expressed in all samples, Vbeta2 and Vbeta16 could be found in 5 samples, whereas Vbeta4-6, Vbeta10-12, Vbeta15, Vbeta17-18, Vbeta20 and Vbeta22-23 were absent in all samples. Genescan analysis showed that clonal expansion of T cells could be found in 1-3 Vbeta subfamilies from 2 cases with B-NHL and 1 case with T-NHL. In conclusions, the similar selected usage of TCR Vbeta subfamily T cells could be found in peripheral blood from patients with B and T NHL, clonal expansion of T cells which were considered to be related to lymphoma cell antigen could be detected in a part of patients with B or T NHL.

The feature of TCR V alpha40 with Jdelta1, Ddelta3 or psiJalpha gene rearrrangements in T cells / 中国实验血液学杂志

The rearrangement segments of TCR Valpha40 gene with Jdelta1, Ddelta3 or psi Jalpha were amplified in genomic DNA from peripheral blood mononuclear cells of 10 normal subjects, sorted CD3(+) cells from peripheral blood of 4 cases and thymocytes from 7 cases, by using nested PCR. Different amounts of DNA from all samples were amplified to estimate the frequency of Valpha40 gene rearrangements. The results indicated that the rearrangements of TCR Valpha40 gene with Jdelta1, Ddelta3 or psi Jalpha could be found respectively in the most samples of peripheral blood T cells or thymocytes. The frequencies of Valpha40 rearrangements were different in peripheral blood T cells and thymocytes by analysis of PCR with different amounts DNA. It is concluded that the TCR V alpha40-psi Jalpha was the most frequent rearrangement in mature and immature T cells, whereas TCR Valpha40-Ddelta3 was more frequently rearranged in immature T cells