The value of cystatin C in evaluating the severity and prognosis of patients with severe fever with thrombocytopenia syndrome.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is a novel emerging viral infectious disease. We explore the value of cystatin C (CysC) level in the evaluation of disease severity and prognosis in patients with SFTS. METHODS: 254 patients with SFTS were enrolled in this study. According to the classification and the outcome of the disease, the patients were divided into the general group and the severe group, the severe patients were divided into the fatal group and the non-fatal group. We compared the laboratory indexes by univariate and multivariate logistic regression analysis to explore the severity and prognostic risk factors of SFTS disease, ROC curve and Kaplan-Meier survival analysis curve were drawn to analyze the independent risk factors and the predictive value of disease severity and prognosis. RESULTS: Univariate analysis showed that the CysC level in severe group and fatal group was significantly higher than general group and non-fatal group (P < 0.05), respectively. Multivariate logistic regression showed that the CysC level was an independent risk factor for severe and death in SFTS patients, and it can effectively predict the risk of severe (AUC = 0.711, 95% CI: 0.645-0.777) and death (AUC = 0.814, 95% CI: 0.737-0.89). The risk of death in patients with cystatin C ≥ 1.23 mg/L was 5.487 times higher than that in patients with cystatin C < 1.23 mg/L. CONCLUSIONS: The CysC level have good predictive value for disease severity and prognosis in patients with SFTS. Trial registration Not applicable.

Longitudinal characteristics of lymphocyte responses and cytokine profiles in the peripheral blood of SARS-CoV-2 infected patients.

BACKGROUND: The dynamic changes of lymphocyte subsets and cytokines profiles of patients with novel coronavirus disease (COVID-19) and their correlation with the disease severity remain unclear. METHODS: Peripheral blood samples were longitudinally collected from 40 confirmed COVID-19 patients and examined for lymphocyte subsets by flow cytometry and cytokine profiles by specific immunoassays. FINDINGS: Of the 40 COVID-19 patients enrolled, 13 severe cases showed significant and sustained decreases in lymphocyte counts [0·6 (0·6-0·8)] but increases in neutrophil counts [4·7 (3·6-5·8)] than 27 mild cases [1.1 (0·8-1·4); 2·0 (1·5-2·9)]. Further analysis demonstrated significant decreases in the counts of T cells, especially CD8+ T cells, as well as increases in IL-6, IL-10, IL-2 and IFN-γ levels in the peripheral blood in the severe cases compared to those in the mild cases. T cell counts and cytokine levels in severe COVID-19 patients who survived the disease gradually recovered at later time points to levels that were comparable to those of the mild cases. Moreover, the neutrophil-to-lymphocyte ratio (NLR) (AUC=0·93) and neutrophil-to-CD8+ T cell ratio (N8R) (AUC =0·94) were identified as powerful prognostic factors affecting the prognosis for severe COVID-19. INTERPRETATION: The degree of lymphopenia and a proinflammatory cytokine storm is higher in severe COVID-19 patients than in mild cases, and is associated with the disease severity. N8R and NLR may serve as a useful prognostic factor for early identification of severe COVID-19 cases. FUNDING: The National Natural Science Foundation of China, the National Science and Technology Major Project, the Health Commission of Hubei Province, Huazhong University of Science and Technology, and the Medical Faculty of the University of Duisburg-Essen and Stiftung Universitaetsmedizin, Hospital Essen, Germany.

The proportion, origin and pro-inflammation roles of low density neutrophils in SFTS disease.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is a novel emerging viral infectious disease. We explored the percentage, origins and functional roles of low density neutrophils (LDNs), one of the neutrophils subsets, in SFTS. METHODS: The LDNs and normal density neutrophils (NDNs) from blood of SFTS and normal volunteers which were collected separately. The percentage, origins and the phagocytic capability of SFTS viral (SFTSV) of LDNs were investigated by flow cytometry and real time PCR. The capacity of LDNs to secrete cytokines and to damage endothelial cells was assessed by ELISA and flow cytometry. RESULTS: We observed that the proportion of LDNs increased dramatically compared with the healthy donors and became the dominant circulating neutrophil population in SFTS patients. Interestingly, the NDNs from the normal donors could switch to LDNs under the SFTS environment. Moreover, SFTSV load in LDNs was significantly higher than that of NDNs in the severe SFTS patients. In addition, the LDNs secreted much higher levels of pro-inflammatory cytokines than NDNs in SFTS and could induce endothelial cell injury. CONCLUSION: The NDNs can be converted to LDNs. This conversion mechanism could become the source of LDNs. The LDNs in severe SFTS patient could engulf more SFTSV and exhibit pro-inflammation functions. TRIAL REGISTRATION: The Ethics Committee of Tongji Medical College, Huazhong University of Science and Technology (IORG No: IORG0003571) gave a final APPROVAL for the study.

Inhibitory effect of miR-125b on hepatitis C virus core protein-induced TLR2/MyD88 signaling in THP-1 cells.

AIM: To investigate the role of miR-125b in regulating monocyte immune responses induced by hepatitis C virus (HCV) core protein. METHODS: Monocytic THP-1 cells were treated with various concentrations of recombinant HCV core protein, and cytokines and miR-125b expression in these cells were analyzed. The requirement of Toll-like receptor 2 (TLR2) or MyD88 gene for HCV core protein-induced immune responses was determined by the transfection of THP-1 cells with gene knockdown vectors expressing either TLR2 siRNA or MyD88 siRNA. The effect of miR-125b overexpression on TLR2/MyD88 signaling was examined by transfecting THP-1 cells with miR-125b mimic RNA oligos. RESULTS: In response to HCV core protein stimulation, cytokine production was up-regulated and miR-125b expression was down-regulated in THP-1 cells. The modulatory effect of HCV core protein on cellular events was dose-dependent and required functional TLR2 or MyD88 gene. Forced miR-125b expression abolished the HCV core protein-induced enhancement of tumor necrosis factor-α, interleukin (IL)-6, and IL-10 expression by 66%, 54%, and 66%, respectively (P < 0.001), by inhibiting MyD88-mediated signaling, including phosphorylation of NF-κBp65, ERK, and P38. CONCLUSION: The inverse correlation between miR-125b and cytokine expression after HCV core challenge suggests that miR-125b may negatively regulate HCV-induced immune responses by targeting TLR2/MyD88 signaling in monocytes.

Association of the Serum Angiotensin II Level with Disease Severity in Severe Fever with Thrombocytopenia Syndrome Patients.

OBJECTIVE: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by a novel Bunyavirus. Recent data suggest that the physiological balance of multiple proinflammatory cytokines is substantially changed in cases of severe fever with thrombocytopenia syndrome virus (SFTSV) infection, and the inflammatory response probably plays an important role in disease progression. Angiotensin II is an important active substance of the renin-angiotensin system, and studies have demonstrated that angiotensin II is involved in key events in the inflammatory process and can regulate inflammatory cell responses. METHODS: In order to elucidate the role of angiotensin II in the pathogenesis of SFTS, we collected serum samples from SFTS patients in the acute or convalescent phase and tested the angiotensin II levels using an enzyme-linked immunosorbent assay as well as SFTSV viral RNA with real-time reverse-transcriptase polymerase chain reaction. Furthermore, we explored possible correlations between the angiotensin II levels and clinical parameters in SFTS patients. RESULTS: Our data showed that the serum level of angiotensin II was significantly increased in the acute phase compared with that seen in the convalescent phase and the healthy controls, while there were no significant differences between the convalescent cases and healthy controls (p>0.05). A correlation analysis demonstrated that the level of angiotensin II positively correlated with the SFTS viral RNA load. The angiotensin II levels were also found to be correlated with clinical parameters indicating impairments in organ functions. Moreover, we also found that the angiotensin II levels were significantly increased in the severe cases versus the non-severe cases (p<0.001). CONCLUSION: The serum angiotensin II levels in SFTS patients may be used to stratify the disease severity and are possibly predictive of disease outcomes.

Decreased monocyte subsets and TLR4-mediated functions in patients with acute severe fever with thrombocytopenia syndrome (SFTS).

OBJECTIVES: The role of a newly discovered bunyavirus, the severe fever with thrombocytopenia syndrome virus (SFTSV), in the pathogenesis of severe fever with thrombocytopenia syndrome (SFTS) is poorly understood. In this study, it was hypothesized that peripheral monocytes, which are constantly exposed to viral infection in the blood, are likely targeted by the causative virus in SFTS patients. METHODS: Fifty-three patients and 25 healthy volunteers were enrolled in the study. Monocyte counts in the peripheral blood of all human subjects were monitored throughout the progress of the disease. SFTSV viral load and the expression of monocyte genes were investigated by real-time RT-PCR. Cytokine production of monocytes in SFTS patients upon lipopolysaccharide (LPS) stimulation was examined by ELISA. RESULTS: In comparison to SFTS patients in the convalescent stage and healthy controls, monocyte cell counts and percentages in patients at the acute stage were significantly lower. Decreased monocyte cell counts and subsets were positively correlated with SFTSV viral loads in the serum samples from SFTS patients. Despite their higher basal toll-like receptor 4 (TLR4) expression, monocytes from patients in the acute phase were shown to be compromised regarding the production of tumor necrosis factor alpha, but not interleukin 10, upon LPS stimulation. CONCLUSIONS: These data strongly suggest that monocytes could be a major target during SFTSV infection. The decreased population and dysfunction of monocytes in acute SFTS patients may contribute to the disease severity.

Abnormal miRNAs Targeting Chromosome Open Reading Frame Genes were Enriched in Microvesicles Derived from the Circulation of HCC.

In this study, we detected the expression profiles of microRNAs (miRNAs) packaged within microvesicles (MVs) from blood samples of HCC patients and healthy donors. Using microarray analysis, there were 83 down-regulated and 92 over-expressed miRNAs in HCC circulation-derived MVs relative to control group. Then potential functions of the dysregulated MVs miRNAs were investigated with bioinformatic tools. We found that 664 Corf genes were targeted by 72 altered MVs miRNAs and some of these target genes were reported to be associated with tumorous activities. Gene Ontology annotation demonstrated that biological roles of the target Corf genes mainly contained the regulation of growth, cell death, macromolecule metabolism, etc. As regulated by abnormal MVs miRNAs, functions of target Corf genes might be interrupted, which were much likely to contribute to HCC occurrence and progression.

Involvement of TLR2-MyD88 in abnormal expression of miR-146a in peripheral blood monocytes of patients with chronic hepatitis C.

miR-146a is an immunoregulatory microRNA closely associated with viral infection. This study investigated the expression changes of miR-146a in peripheral blood monocytes of HCV-infected patients and the mechanism by which the THP-1 cells were stimulated with HCV core protein in vitro. It was found that in the peripheral blood monocytes of HCV-infected patients, miR-146a expression was upregulated. After treated by interferon/ribavirin, miR-146a expression was decreased when HCV RNA became undetectable. HCV core could directly stimulate THP-1 cells to produce miR-146a. Silencing TLR2 and MyD88 could significantly inhibit the expression of miR-146a. It was concluded that the expression of miR-146a in peripheral blood monocytes of HCV-infected patients was abnormally increased. The TLR2-MyD88 signaling pathway may take part in the overexpression of miR-146a in monocytes stimulated with HCV core protein.

Circulating regulatory T cells in patients with severe fever with thrombocytopenia syndrome.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is a newly emerging infectious disease caused by SFTS virus (SFTSV). Immunologic factors have been proved to be related to the occurrence and development of SFTS; however, their role still remains to be further elucidated. METHODS: Samples from 30 patients with laboratory-confirmed SFTS and 15 healthy controls were subjected to flow cytometry to detect the proportion of CD4+/total lymphocytes, CD4 + CD25+/CD4 + cells and CD4 + CD25+ Foxp3+/CD4 + CD25+ cells in circulating blood and to evaluate their potential function in the development of SFTS. RESULTS: The data showed that a reduced proportion of CD4+/total lymphocytes and CD4 + CD25+/CD4 + cells was observed in patients with SFTS compared with healthy controls. In contrast, the percentage of CD4 + CD25+ Foxp3+/CD4 + CD25+ cells in the patients in the SFTS group was significantly elevated. Furthermore, we investigated the dynamic changes of the circulating regulatory T cells (Tregs) in patients with SFTS at different stages. The results showed that the proportion of CD4+/total lymphocytes and CD4 + CD25+/CD4 + cells in the non-severe group was prominently higher than that in patients with severe SFTS. Conversely, the proportion of CD4 + CD25+ Foxp3+/CD4 + CD25+ cells was lower in the non-severe group than in the severe group. Additionally, the circulating Tregs reverted to normal ranges during the convalescent phase of SFTSV infection. Moreover, the Tregs level correlated with various clinical parameters. CONCLUSION: We demonstrated that SFTSV infection resulted in a robust circulating Treg response in patients with SFTS. Our investigation suggested that the proportions of CD4+/total lymphocytes and CD4 + CD25+ Foxp3+/CD4 + CD25+ cells in circulating blood could serve as sensitive indices to evaluate the changes in Tregs in SFTS and predict the progression of SFTS.

Detection and evaluation of immunofunction of patients with severe fever with thrombocytopenia syndrome.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by SFTS virus (SFTSV) with a high fatality rate. But the immunofunction was still unclear. The objective of our study was to assess the immunofunction in SFTS patients. Immunofunction test with flow cytometry which contains CD3+, CD4+ and CD8+ T lymphocytes, B cells and NK cells would be used for detecting serum samples collected from 34 SFTS cases and 20 healthy donors. We found that CD3+ and CD4+ T lymphocytes were significantly diminished in SFTS compared to normal control. In contrast, the percentage of NK cells was elevated. Further analysis revealed that the number of CD3+ and CD4+ T lymphocytes showed that there was a more robust pattern of depression in acute phase and severe SFTS infection compared to the patients in recovery phase and mild SFTS infection. But NK cells were significantly increased in acute phase and severe SFTS. They reverted to the near normal levels in convalescent phase. Additionally, the levels of CD3+ and CD4+ T lymphocytes progressively decreased in death group when compared with the survival group, but the level of B cells was higher. The damages of immune system were obvious, and the immune dysfunction might be partly responsible for disease progression of patients with SFTSV infection.

Comparison of microRNA expression profiles in HCC-derived microvesicles and the parental cells and evaluation of their roles in HCC.

To determine whether the microRNAs (miRNAs) contained in cancer-derived microvesicles (MVs) mirror those of the parental tumor cells, we compared the miRNA expression profiles of MVs derived from their parental hepatocellular carcinoma (HCC) cells. The presence and levels of 888 miRNAs from SMMC-7721 cells and MVs were detected by Agilent miRNA microarray analysis. Four selected miRNAs were verified by real time qRT-PCR. Furthermore, the genes of the miRNAs were bioinformatically identified to explore potential roles of the miRNAs in HCC microenvironment. Our results showed that miRNAs expression profiles of MVs derived from HCC were significantly changed. Of all the miRNAs tested, 148 miRNAs were co-expressed in MVs and SMMC-7721 cells, only 121 and 15 miRNAs were detected in MVs and SMMC-7721 cells, respectively. Among the 148 co-expressing miRNAs, 48 miRNAs had the similar expression level and 6 of them were supposed to be oncogenic or suppressive miRNAs. According to the target prediction by Quantile Algorithm method, these miRNAs may regulate 3831 genes which were closely related to cell cycle, apoptosis and oncogenesis, and 78 were known tumor suppressors or oncogenes. Gene ontology (GO) analysis indicated that 3831 genes were mainly associated with nucleic acid binding, cell death, cell adhesion. MVs containing miRNAs, released into the HCC microenvironment, bear the characteristic miRNAs of the original cells and might participate in cancer progression.

Close correlation between development of MODS during the initial 72 h of hospitalization and hospital mortality in severe fever with thrombocytopenia syndrome.

An emerging infectious disease was identified as severe fever with thrombocytopenia syndrome (SFTS) in central China since late March 2009. We found the patients with SFTS had severe clinical symptoms, and progressed rapidly to multiple organ dysfunction syndrome (MODS) with high fatality rate of 25%-30%. The aim of this study was to assess the significance of risk factors predicting the development of MODS and death in SFTS patients. Consecutive SFTS admissions between May 2009 and September 2011 were analyzed for parameters of organ function during hospitalization using Marshall scoring system for MODS, and platelet counts were recorded on admission and at 24, 48, 72 h and one week after admission. We investigated the kinetics of organ failures and analyzed the association between age, platelet count and development of MODS or death. A total of 92 SFTS patients were enrolled in this study. Among them, 32 patients with dysfunction of over 4 organs were identified, 45% of them died within 72 h, 72% died within 5 days, and 76% died within 7 days after admission. We also found cumulative Marshall score was significantly higher in death patients (11.76±2.05) than in survival patients (4.22±1.98) (P<0.001). In addition, SFTS patients had older age and lower platelet counts in MODS and death groups. Furthermore, we also observed that there was a close correlation between platelet count on admission and Marshall score (P<0.001). High Marshall score, advanced age and lower platelet counts were the main risk factors for the development of MODS, and those factors could predict mortality in SFTS patients, suggesting prompt treatment and close monitoring of severe complications, especially MODS, are of great importance in saving patients' lives.

Peripheral blood microvesicles are potential biomarkers for hepatocellular carcinoma.

BACKGROUND: Microvesicles (MVs) are produced through the outward vesicles budding and fission from the cell surface. Recently, it was discovered that extracellular MVs circulate in bodily fluids of cancer patients and could serve as potential diagnostic biomarkers. However, the diagnostic and prognostic roles of peripheral circulating MVs for hepatocellular carcinoma (HCC) remain unclear. OBJECTIVE: The aim of this study was to investigate whether the peripheral blood MVs could serve as potential biomarkers for detection of HCC. METHODS: Peripheral blood samples were obtained prior to treatment from 55 patients with HCC, 40 patients with liver cirrhosis and 21 healthy controls. MVs were isolated from peripheral blood by centrifugation and measured by using bicinchoninic acid assay. RESULTS: Peripheral blood MVs levels were significantly elevated in HCC patients compared to those in liver cirrhosis (p< 0.001). Furthermore, MVs levels was correlated with the HCC tumor size, pathological classification and TNM stage (p< 0.01). Of note, MVs levels were significantly reduced in the 1 month post-operative blood samples when compared to those in the pre-operative samples in the 17 HCC cases tested. MVs levels did not relate to liver enzymes, AFP levels, alcohol drinking or smoking habits (p> 0.05). In contrast, serum MVs levels correlated with the age of patients, leukocytes, platelets and prothrombin time. The results of receiver operating characteristic (ROC) analysis indicated better performance of MVs than AFP for early detection of HCC. The areas under the ROC curve of MVs for discriminating patients with early (TNM stage I) and relatively early (TNM stage II) HCC from liver cirrhosis was 0.83 (95% CI: 0.74-0.93) and 0.94 (95% CI: 0.88-1.00), respectively. CONCLUSIONS: Peripheral blood MVs levels were increased in patients with HCC and associated with the progression of disease. Serum MVs might serve as novel biomarkers for the diagnosis of HCC at early stage.

MicroRNA expression profiles of circulating microvesicles in hepatocellular carcinoma.

BACKGROUND: Microvesicles (MVs) packaged with microribonucleic acids (miRNAs) have been shown to be released mainly from tumor cells. However, little information is known for miRNAs from MVs in hepatocellular carcinoma (HCC). Hence, we explored the MVs' miRNAs expression profiles in HCC. METHODS: MVs were collected from peripheral blood of HCC, chronic hepatitis B (CHB) and normal controls. miRNA from MVs were labeled and analyzed by Agilent miRNA microarry. Bioinformatics tools were used to analyze function of the differentially expressed MVs' miRNAs. RESULTS: A total of 242 aberrantly expressed miRNAs were identified in HCC-MVs compared with CHB-MVs and the control. Among them, 115 miRNAs were over-expressed with up to 31 fold difference (miR-671-5p) and 127 were down-expressed with up to 0.041 fold difference (miR-432) in HCC. By software miRror2.0, nucleolar protein 3 (NOL3) was found to be the core player among 300 target genes of top ten up-regulated miRNAs and serine/ arginine repetitive matrix 1(SRRM1) was central among the 219 targets of the top ten down-regulated miRNAs. We also analyzed GO categories for these predicted genes : cellular component, biological processes, and molecular function. The deregulation of MVs' miRNAs and their target genes were closely involved in the pathways of HCC. CONCLUSIONS: Our study firstly demonstrated that miRNAs were differentially expressed in HCC-MVs compared with CHB and normal controls. Aberrant HCC-MVs miRNAs may play important roles in the development of HCC.

The clinical characteristics and outcomes of patients with human granulocytic anaplasmosis in China.

OBJECTIVES: The incidence of human granulocytic anaplasmosis (HGA), a tick-borne disease caused by the obligate intracellular bacterium Anaplasma phagocytophilum, has increased across the world. However, information on HGA is lacking in China. The purpose of this study was to investigate the clinical features and outcomes of HGA patients in China. METHODS: A total of 83 patients with HGA from the provinces of Hubei and Henan in China, who were admitted to Union Hospital between March 2009 and September 2010, were included in this study. We investigated the epidemiology, clinical features, laboratory markers, and therapeutic effects in these patients. We also analyzed life-threatening complications such as systemic inflammatory response syndrome (SIRS)/multiple organ dysfunction syndrome (MODS) following HGA and assessed the risk factors for a poor clinical outcome. RESULTS: In our study, an HGA outbreak peak was observed for the months May to August. The highest age-specific incidence occurred among the group of patients aged 50-59 years. With regard to patient occupation and pathological origin, we found that 73 of the 83 patients with HGA had a peasant occupation. With respect to symptoms, 45 patients had no complications and 38 patients diagnosed with HGA met SIRS criteria, of whom 25 rapidly developed MODS. The mortality for the entire cohort was 26.5%. The factors predictive of patients developing MODS and an adverse outcome were advanced age, disturbance of consciousness, highly elevated lactate dehydrogenase, creatinine, and aspartate aminotransferase levels, and the presence of SIRS. Moreover, MODS was found to be an independent predictor of death. CONCLUSIONS: In China, HGA patients had severe clinical symptoms and high rates of complications and mortality. These findings may provide useful information so that physicians will be on the alert for severe complications after a diagnosis of HGA; they will also be useful for optimizing supportive care for HGA-related critical illness. Prompt treatment and close monitoring of severe complications such as SIRS and MODS are of great importance in saving patient lives.

Circulating LTD4 in patients with hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a type of inflammation-related cancer that usually follows chronic inflammations. Leukotriene D4 (LTD4) is a potent biologically active arachidonic acid-derived lipid mediator that is intimately involved in inflammations and cancers. Although previous researches found overexpression of LTD4 in several other cancers, the circulating LTD4 level in HCC remains unknown. The aim of this study was to examine concentrations of LTD4 and analyze its roles in HCC. The results showed that remarkably high circulating LTD4 in HCC versus healthy subjects (p < 0.001). The levels of LTD4 were neither associated with parameters expressing tumor burden, such as AFP, nor with inflammation factors AST and γ-GT. In addition, the significant increase of circulating LTD4 levels was obtained in patients with HCC accompanied by chronic hepatitis B (CHB), compared with those patients suffering HCC alone(P < 0.05). Furthermore, although the slightly lower levels of LTD4 were detected in HCC patients with non-metastasis and therapy compared with metastasis and non-therapy, no significant differences were detected. Taken together, the levels of circulating LTD4 are elevated in HCC and it may participate in the pathogenesis of HCC as an inflammatory factor from CHB disease to HCC.

Berberine inhibits angiogenic potential of Hep G2 cell line through VEGF down-regulation in vitro.

BACKGROUND AND AIM: Berberine, an herbal alkaloid, has been reported to have promotion potential of apoptosis and anticancer effect on a variety of human tumor cells. To obtain more specific understanding of those consequences of berberine on hepatocellular carcinoma (HCC) and the tumor microenvironment, we conducted in vitro experiments to investigate the inhibitory effect of berberine on tumor-induced angiogenesis using HCC cells and human umbilical vein endothelial cells (HUVECs). METHODS: Human umbilical vein endothelial cell growth was quantified with the CCK-8 cell proliferation assay; cell migration was observed with a Boyden chamber (Transwell, Corning, Lowell, MA, USA), and angiogenesis was assessed by endothelial tube formation in Matrigel in vitro. In addition, VEGF level was determined by ELISA and VEGF mRNA expression by RT-PCR. RESULTS: Berberine inhibited the capacity of HCC to stimulate HUVEC's proliferation, migration and endothelial tube formation, suggesting that berberine could influence the cross-talk between the HCC cell and vascular endothelial cells. These results demonstrate berberine's antiangiogenesis property and its clinical potential as an inhibitor of tumor angiogenesis. Subsequently analyses reveal that berberine prevents secretion of VEGF from HCC and down-regulates VEGF mRNA expression. CONCLUSION: These findings strongly suggest that berberine is a potential antiangiogenic agent and a promising antitumor drug for HCC.

The role of hERG1 K+ channels and a functional link between hERG1 K+ channels and SDF-1 in acute leukemic cell migration.

Stromal cell-derived factor-1 (SDF-1) and its unique receptor, CXCR4, regulate stem/progenitor cell migration and retention in the bone marrow and are required for hematopoiesis. Recent studies found that hERG1 K(+) channels were important regulators of tumor cell migration. In this study, we investigated whether SDF-1 induced acute leukemic cell migration associated with hERG1 K(+) channels. Our results showed that E-4031, a specific hERG1 K(+) channels inhibitor, significantly blocked SDF-1-induced migration of leukemic cell lines, primary acute leukemic cells, leukemic stem cells and HEK293T cells transfected with herg-pEGFP. The migration of phenotypically recognizable subsets gave the indication that lymphoblastic leukemic cells were inhibited more than myeloid cells while in the presence of E-4031 which maybe associated with herg expression. SDF-1 increased hERG1 K(+) current expressed in oocytes and HEK293T cells transfected with herg-pEGFP. There were no significant changes of CXCR4 expression on both HL-60 cells and primary leukemic cells regardless if untreated or treated with E-4031 for 24 h (P>0.05). The hERG1 K(+) current increased by SDF-1 might contribute to the mechanism of SDF-1-induced leukemic cell migration. The data suggested that hERG1 K(+) channels functionally linked to cell migration induced by SDF-1.

Tea polyphenols exerts anti-hepatitis B virus effects in a stably HBV-transfected cell line.

In this study, the anti-HBV effects of tea polyphenols (TP) were examined. After cells were exposed to TP for 3, 6, 9 days, amounts of HBsAg, HBeAg and HBV-DNA released into the supernatant of the cultured HepG2 2.2.15 cells were detected. TP, to some extent, inhibited the secretion of HBsAg and strongly suppressed the secretion of HBeAg in a dose-dependent (P<0.01) and time-dependent manner, with 50% maximal inhibitory concentration (IC50) value being 7.34 microg/mL on the 9th day, but the time-dependence was not significant (P=0.051). Expression of HBV-DNA in the supernatant of the cell culture also was significantly decreased in a dose-dependent fashion (P<0.01). The IC50 of TP in inhibiting HBV DNA was 2.54 microg/mL. It concluded that TP possessed potential anti-HBV effects and may be used as a treatment alternative for HBV infection.

Tea Polyphenols Exerts Anti-hepatitis B Virus Effects in a Stably HBV-transfected Cell Line / 华中科技大学学报（医学）（英德文版）

In this study,the anti-HBV effects of tea polyphenols (TP) were examined.After cells were exposed to TP for 3,6,9 days,amounts of HBsAg,HBeAg and HBV-DNA released into the supernatant of the cultured HepG2 2.2.15 cells were detected.TP,to some extent,inhibited the secre-tion of HBsAg and strongly suppressed the secretion of HBeAg in a dose-dependent (P<0.01) and time-dependent manner,with 50% maximal inhibitory concentration (IC50) value being 7.34 μg/mL on the 9th day,but the time-dependence was not significant (P=0.051).Expression of HBV-DNA in the supernatant of the cell culture also was significantly decreased in a dose-dependent fashion (P<0.01).The IC50 of TP in inhibiting HBV DNA was 2.54 μg/mL.It concluded that TP possessed potential anti-HBV effects and may be used as a treatment alternative for HBV infection.