Objective To compare the effecacy and safety of lower dose peginterferon alfa-2b plus ribavirin versus standard interferon alfa-2b plus ribavirin for treatment of chronic hepatitis C in China.Methods 192 patients with chronic hepatitis C were assigned peginterferon alfa-2b 0.5?g/kg each week plus ribavirin 750～1050 mg/d or standard interferon alfa-2b 3 MIU TIW plus ribavirin 750～1050 mg/d for 48-week treatment and 24-week fellow-up.Results The sustained virological response (SVR)rate in lowe-dose peginterferon alfa-2b plus ribavirin group was 53.8%,and the SVR rate in standard interferon alfa-2b plus ribavirin was 58.1%.The SVR was similar between the two treat- ment group(P=0.966 for both comparisons).The adverse event rate was higher in peginterferon al- fa-2b plus ribavirin group than in standard interferon alfa-2b plus ribavirin group(P=0.033 for both comparisons),but there was no new or unique adverse event in related to pegylation of interferon alfa-2b. Conclusion The effecacy and safety was similar between lower-dose peginterferon alfa-2b plus ribavirin and standard interferon alfa-2b plus ribavirin for treatment of chronic hepatitis C.

Objective To explore the effects of overexpression of hypoxia inducible factor 1? (HIF 1?) mRNA on vascular endothelial growth factor (VEGF) and novoendotheliasis in venous autografts Methods Wistar rats were randomly divided into two groups with 28 rats in each group A rat experimental model of autogenous vein graft was established by transplanting the right external jugular vein into between the interrupted right common carotid artery The transplanted vein in the experimental group was first immersed into a solution containing recombinant adeno associated virus (rAAV) HIF 1? for 45 minutes Vein grafts and blood simples were taken at 7 or 14 days after transplantation RT PCR, ELISA, immunohistochemistry were used to detected HIF 1? mRNA and VEGF expression Results HIF 1? mRNA and VEGF protein remarkably increased in experimental group, and serum level of E selectin significantly decraesed at day 14 The novoendotheliasis and myo endothelium junction in vein grafts were remodeled at day 14 in the experimental group Conclusion Re establishment of the structure and function of the autograft vein graft endothelium was accelerated by overexpressed HIF 1? mRNA

To study the expression of HCV non-structure 5 antigen in vitro, a human HepG2 cell line was incubated with a HCV RNA positive serum. The S ABC i mmunological techniques and gold-labeled colloid electron microscopy method wer e employed to examine for the viral proteins in those cells. The HCV non-struct ure 5 antigen was first detected in the HepG2 cells at 72 hours post incubation. The antigen was continuously observed in the cytoplasm or on the membrane as we ll on the cell wall of the HepG2 cells even after 1, 2, 3 and 4 weeks post incub ation. The observation of HCV non-structure 5 antigen continuously expressed in the HepG2 cells strongly indicates that the cells may have been infected by HCV virus and the virus may have replicated in the cells. Therefore, the HepG2 cell line may be served as a potential host for establishment of HCV infection and p ropagation in vitro.

OBJECTIVETo establish hepatitis C virus (HCV) infected cell model which is similar to the infection in vivo and can support HCV to replicate for a long time.

METHODSAfter infected with HCV-positive serum, Hep G2 cells were cultured for 60 days. Nested RT-PCR was used to detect plus and minus HCV RNA in cultured cells and supernatants.

RESULTSPlus HCV RNA was detected intermittently in Hep G2 cells during 2-30 days, minus HCV RNA was detected during 3-30 days after infection, the detection rate was similar to plus HCV RNA. Plus and minus HCV RNA can be still intermittently detected during 31-60 days after infection. However, the detection rate gradually declined. Plus HCV RNA was also found intermittently positive in the supernatant, and the detection rate was consistent to that in cells. Minus HCV RNA was not detected in the supernatant.

CONCLUSIONHep G2 cells were susceptible to HCV, and could support HCV to replicate for a relatively long time. Hep G2 is an ideal HCV infection cell model.

Cell Line, Tumor ; Hepacivirus ; genetics ; growth & development ; Humans ; RNA, Viral ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Virus Replication

Animals ; Female ; Liver Cirrhosis, Experimental ; drug therapy ; parasitology ; Mice ; Mice, Inbred Strains ; Pentoxifylline ; therapeutic use ; Schistosomiasis japonica ; drug therapy

This study was purposed to investigate the therapeutic effects of early transfusion of immunized donor lymphocytes after haploidentical transplantation by means of mouse model of nonmyeloablative haploidentical bone marrow transplantation. CB6F1 female mouse was served as recipient and C57BL/6 male mouse was served as donor. Each CB6F1 female mouse was subjected to intravenous transfusion with 1×10(6) erythroleukemia (EL9611) cells at day 4 before transplantation, followed with intraperitoneal injection of Ara-C (0.015 g) respectively at day 2 and day 1, then conditioned for BMT with TBI (450 cGy) at day 1 before transplantation. After conditioning (day 0), each of recipients was transplanted with 6×10(7) mixture of bone marrow and spleen cells from the C57BL/6 mice, and was infused with 6 × 10(7) immunized donor lymphocytes at day 15 after transplantation. All treated animals were evaluated for survival, development of leukemia and aGVHD. The donor CD3(+) cell chimerism and sex determining region Y gene (SRY)in recipients were monitored periodically after transplantation. The results showed tht all mice with only inoculation of 10(6) EL9611 cells survived for 15 ± 1 days (n = 4); all mice of other groups obtained the varying degrees of implantation. SRY could be detected at day 30 and 60 after transplantation. The chimerism of donor CD3(+) cells in mixed bone marrow transplantation (MT) group at day 14, 30 and 60 respectively reached 17.95% ± 12.03%, 37.34% ± 2.78% and 47.06% ± 6.1%. In donor lymphocyte infusion (DLI) group it reached 69.78% ± 12.62%, 75% ± 15.97%, 83.41% ± 16.07% at day 30, 45 and 60 after transplantation. The mice of MT and DLI group survived for 66.66 ± 1.47 days and 78.2 ± 7.82 days. It is concluded that the high tumor burden before transplantation can affect donor cell engraftment and prognosis.Early post-transplanted infusion of immunized lymphocytes from donor can help to improve the therapeutic efficacy and survival.
Animals ; Bone Marrow Transplantation ; methods ; Female ; Haplotypes ; Leukemia, Erythroblastic, Acute ; therapy ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors ; Transplantation Conditioning ; methods ; Transplantation, Homologous

OBJECTIVETo detect the expression profile of NK ligands in acute leukemia cell lines and investigate the differential expression pattern between acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).

METHODSUsing quantitative real-time PCR, 23 NK ligands (MICA, MICB, ULBP-1, ULBP-2, ULBP-3, ULBP-4, HLA-E, HLA-G, CD48, NBTA, HLA-F, LLT-1, PVR, Nectin2, CD72, CD80, ICAM-1, LFA-3, CRACC, Fas, DR4, DR5, TNFR1) were detected in 6 acute leukemia cell lines, including 3 ALL cell lines (CEM, Jurkat T, Reh) and 3 AML cell lines (HL-60, KG-1a, NB4), respectively. Independent-samples t test analysis was performed to determine statistical significance.

RESULTSUsing β-actin as reference gene, the relative expression results showed that the expression of 4 NK ligands between ALL and AML is significantly different. Specifically, the level of ULBP-2 is higher in ALL (CEM: 1, Jurkat T: 0.617, Reh: 0.246) than that in AML (HL-60: 0.000, KG-1a: 0.003, NB4: 0.000)(P = 0.047). However, the expressions of CD48, PVR(PVR-1, PVR-2) and DR4 is higher in AML (HL-60: 13.987, 4.403, 10.334, 8.711; KG-1a: 5.387, 2.900, 7.315, 4.512; NB4: 7.763, 3.248, 7.049, 6.127) than that in ALL (CEM: 1, 1, 1, 1; Jurkat T: 2.035, 1.553, 3.888, 0.449; Reh: 1.559, 0.000, 0.000, 1.304) (P = 0.044, 0.014, 0.014, 0.011). And there're no significant differences between the rest 19 NK ligands.

CONCLUSIONSULBP-2, CD48, PVR and DR4 might play an important role in the distinct mechanisms in leukemogenesis between ALL and AML and could be potential targets for diagnosis and treatment.

Acute Disease ; Antigens, CD ; genetics ; metabolism ; CD48 Antigen ; Cell Line, Tumor ; GPI-Linked Proteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Ligands ; Membrane Proteins ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Receptors, Virus ; genetics ; metabolism

OBJECTIVETo establish a new and efficient method(IEDA) for detection of hemorrhagic fever with renal syndrome virus (HFRSV) antigen.

METHODSAn immune enzyme dot assay (IEDA) with mixture of three sorts anti-HFRSV-IgG, which was obtained from rabbit vaccinated with EHFV R22, Chen and Hubei strain was employed to detect HFRSV antigen in serum and urine from epidemic hemorrhagic fever (EHF) patients, and compared with indirect immune fluorescence assay (I-IFA), 76 serum samples and 40 urine samples were detected in this study.

RESULTSThe results showed that the total positive rate of HFRSV antigen detected by IEDA was 73.68% in serum and 65.00% in urine, while that detected by I-IFA was 75.00% and 70.00%, respectively. The positive rate in primary phase (within 5 days) of HFRSV antigen detected by IEDA was 94.34% in serum and 83.33% in urine, while that detected by I-IFA was 64.42% and 55.56%, respectively, there was significant difference in both serum and urine detections. Correlation study showed a high correlation in the result of IEDA and I-IFA.

CONCLUSIONThe results of this study suggested that the IEDA, as compared with I-IFA, was a more specific, sensitive, rapid and simple method with higher positive rate in primary phase. IEDA could be widely used for early diagnosis of HFRS in hospital at grassroots level.

Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; analysis ; Early Diagnosis ; Female ; Fluorescent Antibody Technique, Indirect ; Hantaan virus ; immunology ; Hemorrhagic Fever with Renal Syndrome ; diagnosis ; immunology ; Humans ; Immunoblotting ; methods ; Immunoglobulin G ; immunology ; Male ; Rabbits ; Rats ; Sensitivity and Specificity

OBJECTIVECoronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome (ACS). Metalloprotease (MMPs) secreted by monocyte/macrophage was the main predisposing factor of the plaque rupture and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-gamma in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS.

METHODSPeripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony-stimulating factor (0.1 microg/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 micromol/L) for 48 hours. The concentrations of MMP-9 and TIMP-1 in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-gamma, MMP-9 by RT-PCR and nuclear factor-kappaB P65 (NF-kappaB P65) expression by immunohistochemistry.

RESULTSPPAR-gamma mRNA expression was significantly lower while NF-kappaB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group (all P < 0.05). After rosiglitazone intervention, PPAR-gamma mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulation of NF-kappaB P65 expression in both groups.

CONCLUSIONRosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-gamma expression, and by downregulating NF-kappaB expression in MDMs isolated from patients with ACS.

Acute Coronary Syndrome ; blood ; Aged ; Case-Control Studies ; Cells, Cultured ; Female ; Humans ; Macrophages ; metabolism ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; PPAR gamma ; agonists ; Thiazolidinediones ; pharmacology ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Transcription Factor RelA ; metabolism ; Vasodilator Agents ; pharmacology

BACKGROUNDH3N2 subtype influenza A viruses have been identified in humans worldwide, raising concerns about their pandemic potential and prompting the development of candidate vaccines to protect humans against this subtype of influenza A virus. The aim of this study was to establish a system for rescuing of a cold-adapted high-yielding H3N2 subtype human influenza virus by reverse genetics.

METHODSIn order to generate better and safer vaccine candidate viruses, a cold-adapted high yielding reassortant H3N2 influenza A virus was genetically constructed by reverse genetics and was designated as rgAA-H3N2. The rgAA-H3N2 virus contained HA and NA genes from an epidemic strain A/Wisconsin/67/2005 (H3N2) in a background of internal genes derived from the master donor viruses (MDV), cold-adapted (ca), temperature sensitive (ts), live attenuated influenza virus strain A/Ann Arbor/6/60 (MDV-A).

RESULTSIn this presentation, the virus HA titer of rgAA-H3N2 in the allantoic fluid from infected embryonated eggs was as high as 1:1024. A fluorescent focus assay (FFU) was performed 24-36 hours post-infection using a specific antibody and bright staining was used for determining the virus titer. The allantoic fluid containing the recovered influenza virus was analyzed in a hemagglutination inhibition (HI) test and the specific inhibition was found.

CONCLUSIONThe results mentioned above demonstrated that cold-adapted, attenuated reassortant H3N2 subtype influenza A virus was successfully generated, which laid a good foundation for the further related research.

Animals ; COS Cells ; Cercopithecus aethiops ; Dogs ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; genetics ; Plasmids ; Reassortant Viruses ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, Attenuated ; immunology ; Viral Proteins ; genetics