Exploring the Role of Epicardial Adipose Tissue in Coronary Artery Disease From the Difference of Gene Expression.

OBJECTIVES: Epicardial adipose tissue (EAT) is closely adjacent to the coronary arteries and myocardium, its role as an endocrine organ to affect the pathophysiological processes of the coronary arteries and myocardium has been increasingly recognized. However, the specific gene expression profiles of EAT in coronary artery disease (CAD) has not been well characterized. Our aim was to investigate the role of EAT in CAD at the gene level. METHODS: Here, we compared the histological and gene expression difference of EAT between CAD and non-CAD. We investigated the gene expression profiles in the EAT of patients with CAD through the high-throughput RNA sequencing. We performed bioinformatics analysis such as functional enrichment analysis and protein-protein interaction network construction to obtain and verify the hub differentially expressed genes (DEGs) in the EAT of CAD. RESULTS: Our results showed that the size of epicardial adipocytes in the CAD group was larger than in the control group. Our findings on the EAT gene expression profiles of CAD showed a total of 747 DEGs (fold change >2, p value <0.05). The enrichment analysis of DEGs showed that more pro-inflammatory and immunological genes and pathways were involved in CAD. Ten hub DEGs (GNG3, MCHR1, BDKRB1, MCHR2, CXCL8, CXCR5, CCR8, CCL4L1, TAS2R10, and TAS2R41) were identified. CONCLUSION: Epicardial adipose tissue in CAD shows unique gene expression profiles and may act as key regulators in the CAD pathological process.

Transfusion of multi-factors activated immune cells as a novel treatment for patients with chronic hepatitis B.

BACKGROUND AND OBJECTIVES: Host-virus interactions play a central role in determining the prognosis of hepatitis B virus infection. Multi-factors activated immune cells (MAICs) are autologous peripheral blood mononuclear cells activated with anti-CD3 monoclonal antibody, interleukin-2 and interferon-gamma. The present pilot clinical trial was designed to determine whether adoptively transferred MAICs inhibit the replication of hepatitis B virus and is safe for patients. STUDY DESIGN: Fourteen patients with chronic hepatitis B were enrolled in the study. A total of (1.5-4.0)x10(9) peripheral blood mononuclear cells were isolated from each patient and activated by anti-CD3 monoclonal antibody, interleukin-2 and interferon-gamma in vitro for 10 days to produce MAICs. Cell phenotypes and levels of cytokine secretion were determined during cell culture. Patients were followed up for 1 year after the transfusion of MAICs. RESULTS: After 10 days of culture, peripheral blood mononuclear cells were expanded to a mean fold of 4.68+/-1.78 and activated effectively, as demonstrated by CD25 expression and cytokine secretion. Cell activation peaked on day 4. All patients accepted the MAICs transfusion with some slight adverse events, and fulminant hepatitis and bilirubinemia were not observed. Significant HBV inhibition was observed in 8 out of 14 patients, in which 5 patients achieved complete response (defined as that serum HBV DNA levels below the detection limit, occurrence of HBeAg seroconversion and normalization of ALT were observed together after MAICs transfusion and kept for at least 6 months) and 3 patients achieved partial response (defined as that serum HBV DNA levels below the detection limit, occurrence of HBeAg disappearance and normalization of ALT were observed together in 6 months after MAICs transfusion, but kept for less than 6 months). CONCLUSION: These findings strongly suggest that MAICs transfusion, which effectively inhibits the replication of hepatitis B virus, is safe for patients.

Dendritic cells from chronic hepatitis B patients can induce HBV antigen-specific T cell responses.

AIM: To determine whether dendritic cells (DCs) from chronic hepatitis B patients could induce HBV antigen-specific T cell responses or not. METHODS: DCs were generated from peripheral blood mononuclear cells of patients with chronic hepatitis B (CHB) infection and healthy donors. We compared the phenotypes of these DCs and their ability to secrete cytokines and to participate in mixed lymphocyte reactions. In addition, autologous lymphocytes were cultured with DCs loaded with HBV core region peptide HBcAg8-27, an epitope recognized by cytotoxic T lymphocytes (CTL), and bearing human leucocyte antigen (HLA)-A2 for 10 d. Cytokine secretion and lytic activity against peptide-pulsed target cells were assessed. RESULTS: DCs with typical morphology were generated successfully by culturing peripheral blood mononuclear cells (PBMCs) from CHB patients with AIM-V containing GM-CSF and IL-4. Compared with DCs from normal donors, the level of CD80 expressed in DCs from CHB patients was lower, and DCs from patients had lower capacity of stimulate T cell proliferation. When PBMCs isolated from patients with chronic or acute hepatitis B infection and from normal donors were cocultured with HBcAg18-27 peptide, the antigen-specific memory response of PBMCs from acute hepatitis B patients was stronger than that of PBMCs from chronic hepatitis B patients or normal donors. PBMCs cocultured with DCs treated with HBcAg18-27 CTL epitope peptide induced an antigen-specific T cell reaction, in which the level of secreted cytokines and lytic activity were higher than those produced by memory T cells. CONCLUSION: DCs from patients with CHB can induce HBV antigen-specific T cell reactions, including secretion of cytokines essential for HBV clearance and for killing cells infected with HBV.

[Impaired non-viral specific immune function of dendritic cell does not interfere with clearance and cytotoxic T lymphocyte response to HBV or HCV].

OBJECTIVE: To investigate the correlation between impaired non-viral specific immune function of dendritic cell (DC) and viral clearance and cytotoxic T lymphocyte (CTL) response to HBV or HCV in patients with HBV and HCV coinfection. METHODS: Twenty-five patients with HBV and HCV coinfection were investigated in this study. In 1994 and 2002, biochemical and virological markers and quantitative serum HBV DNA and HCV RNA levels were detected in these patients. According to the virus clearance status, these patients were divided into 4 groups: 14 patients with both HBV and HCV clearance (Group A), 6 patients with HCV clearance only (Group B), 3 patients with HBV clearance only (Group C), and 2 patients with persistent infection of HBV and HCV (Group D). Phenotypes and immune functions of monocyte-derived DCs were compared between these groups. 51Cr release assay were used to measure CTL response to epitopes derived from HBV, HCV or influenza virus (as positive control) in HLA-A2+ patients. RESULTS: Impaired non-viral specific immune functions of DCs were observed in group B, C and D compared with group A and normal donors (Group N). These impaired functions included CD86 decreasing expression and lower capacity to stimulating allogenic T cells and uptaking antigen. The specific CTL response to HBV- and HCV-derived peptides could be induced in group A (12/12). The specific CTL response to HBV-derived peptides or to HCV-derived peptides could be induced in group C (3/3) or B (5/5), respectively. But the specific CTL response to both of two HBV-derived peptides or two HCV-derived peptides could not be induced in group C (0/3) or B (0/5), respectively. And no CTL response to HBV or HCV-derived peptides could be induced in groups D (0/1) and N (0/4). CONCLUSION: 1. The results suggest that specific CTL response to HBV or HCV play a vital role in the viral clearance. 2. The DCs with impaired non-viral specific immune functions exist in chronic patients with HBV and/or HCV infection, but do not interfere with clearance and CTL response to HBV or HCV. It is reasonable to speculate that impaired functions of DCs result from viral infection.

[Dendritic cells originated from the peripheral blood in chronic hepatitis B patients can induce specific T cell immune response].

OBJECTIVE: To study whether dendritic cells (DCs) derived from the peripheral blood in chronic hepatitis B patients can induce specific T cell immune response. METHODS: (1)The subjects were divided into 3 groups: chronic hepatitis B group (CHB), acute hepatitis B group (AHB), and normal donor group (ND). The peripheral blood mononuclear cells (PBMCs) isolated from those subjects were stimulated with HBcAg 18 to 27 CTL epitope peptide, and intracellular cytokine staining (ICCS) was used for detecting IFN-gamma, IL-2 and TNF-alpha produced by CD8+ T cell. (2) DCs generated from PBMCs were pulsed with HBcAg 18 to 27 CTL epitope peptide, then were cocultured with autologous lymphocytes for 10 days to induce antigen-specific T cell, which was assessed by ICCS and cytotoxic assay. RESULTS: (1) The memory effect of the PBMCs from AHB group to HBcAg 18 to 27 CTL epitope peptide was stronger than that from CHB or ND group (t=2.508-3.305, P<0.05). (2)After lymphocytes were cocultured with DC treated with HBcAg 18 to 27 CTL epitope peptide, antigen-specific T cell effect was induced. And the killing rates were (57.0+/-23.0)%, (49.5+/-20.2)%, (21.8+/-12.9)% at the effector/target of 30:1, 10:1, 3:1, which were higher than that in control group. CONCLUSIONS: The memory T cells against HBV antigen lacks in CHB patients. DCs from CHB patients pulsed with HBcAg 18 to 27 epitope peptide can induce HBV antigen-specific T cell, which can kill specific target cells and produce cytokines involved in virus clearance.

Spontaneous viral clearance after 6-21 years of hepatitis B and C viruses coinfection in high HBV endemic area.

AIM: To investigate the clinical and virological course of coinfection by hepatitis B virus (HBV) and hepatitis C virus (HCV) in China. METHODS: We enrolled 40 patients with chronic HBV and HCV coinfection (Group BC), 16 patients with chronic HBV infection (Group B) and 31 patients with chronic HCV infection (Group C). They infected HBV and/or HCV during 1982 to 1989. Sera of all the 87 patients were collected in 1994 and 2002 respectively. We detected biochemical and virologic markers and serum HBV DNA and HCV RNA levels of all the patients. B-type ultrasound detection was performed in some patients. RESULTS: In Group BC, 67.5 % of the patients cleared HBsAg, and 92.5 % of the patients cleared HBeAg. The clearance rate of HBV DNA was 87.5 %. There was no significant difference of HBV clearance between Group BC and Group B. In Group BC, 85.7 % of males and 47.4 % of females cleared HBV, and males were easier to clear HBV (chi(2)=6.686, P=0.010). Such a tendency was also found in Group B. The clearance rate of HCV RNA in Group BC was 87.5 %, significantly higher than that in Group C (chi(2)=22.963, P<0.001). Less than 40 % of the patients in all groups had elevated liver enzyme values. The highest value of alanine aminotransferase (ALT) was 218 u/L (normal range for ALT is 0-40 u/L). In most patients the ultrasonogram presentations changed mildly. CONCLUSION: The clinical manifestations of patients with HBV/HCV coinfection are mild and occult. High clearance rate of HBV and easy to clear HBV in male patients are the characteristics of HBV infection in adults in China. HBV can inhibit HCV replication, but no evidence has been found in our data that HCV suppresses HBV replication.

[Clinical and virological course of dual infection by hepatitis B and C viruses in China].

OBJECTIVE: To investigate the clinical and virological course of dual infection by hepatitis B virus (HBV) and hepatitis C virus (HCV) in China. METHODS: We enrolled 40 patients with chronic HBV and HCV dual infection (Group BC), 16 patients with chronic single HBV infection (Group B) and 31 patients with chronic single HCV infection (Group C). They infected HBV and/or HCV during 1982 to 1989. The sera of all the 87 patients were collected in 1994 and 2002 respectively. We detected biochemical and virologic markers and quantitative serum HCV DNA and HCV RNA levels of all the patients. The B-type ultrasound detecting was performed in some patients. RESULTS: In Group BC, 67.5% of the patients have cleared HBsAg, and 92.5% of the patients have cleared HBeAg. The clearance rate of HBV DNA was 87.5%. There was no significant difference of HBV clearance between Group BC and Group B. In Group BC, the clearance rate of HBV-RNA was 87.5%, significantly higher than those in Group C. Less than 40% of the patients in all groups have elevated liver enzyme values. The highest value of alanine aminotransferase (ALT) was 218 u/L (normal range for ALT 0 - 40 u/L). In most patients the ultrasonogram presentations were changed mildly. CONCLUSION: The clinical presentation of patients with HBV/HCV coinfection seemed mild and occult. High clearance of HBV, easy to clear HBV in male patient was the characteristics of infection by HBV in adult time in China. HBV could inhibit HCV replication, but no evidence could be found in our data that HCV could suppress HBV replication.

Tissue-engineered heart valve on acellular aortic valve scaffold: in-vivo study.

The feasibility of constructing a tissue-engineered heart valve on an acellular porcine aortic valve leaflet was evaluated. A detergent and enzymatic extraction process was developed to remove the cellular components from porcine aortic valves. The acellular valve leaflets were seeded for 7 days in vitro with cells from canine arterial wall and endothelial cells. The constructs were implanted into the lumens of 6 canine abdominal aortas to assess the reconstruction of the valve leaflets. It was found that all cellular components had been removed from the porcine aortic valves. The valve leaflets were completely reconstructed at the end of the 10th week in vivo. Scanning electron microscopy showed that the valve leaflets were partially covered with endothelial cells. It was concluded that porcine aortic valves can be decellularized by the detergent and enzymatic extraction process and it is feasible to construct a tissue-engineered heart valve in vivo on an acellular valve scaffold.

[Comparison of hepatitis B virus serotype and genotype among HBsAg positive hepatitis B patients in a northern and a southern city of China].

OBJECTIVE: To understand HBV serotypes and genotypes epidemiology in a northern city and a southern city in China. METHODS: Using polymerase chain reaction (PCR) and direct sequencing of HBV DNA PCR products, the serotypes and genotypes of HBV in 530 from HBsAg positive samples. The enrolled patients were from Harbin, a northern city and Lianjiang, a southern city in China. RESULTS: Comparison of the serotypes and genotypes of HBV between Harbin and Lianjiang showed that adrq+ was the most predominant hepatitis B virus serotype in both Harbin and Lianjiang (87.2% and 73.5%,respectively), adw2 was the next (12.0% and 25.7%, respectively); genotype C was the most frequent in Harbin and Lianjiang (87.8% and 73.2%, respectively), and genotype B was the next (12.2% and 26.1%, respectively) only 1 patient was infected by genotype D, and 1 patient was found to be co-infected by genotype B and C in Lianjiang. CONCLUSION: The results suggest that the percentage of HBV serotypes and genotypes between Harbin and Lianjiang was significantly different (P less than 0.001), but the main HBV serotype and genotype of the two cities were similar.