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Biomedical subjects

Fu-sheng Wang

Publications and source records attributed to Fu-sheng Wang.

30 records · Page 2Linked to original sources
PubMed2003

[Identification of immunological effector cells after autologous cytokine-induced killer cells treatment and its clinical implication in hepatocellular carcinoma patients].

OBJECTIVE: To investigate the alteration of the cellular profiles of T lymphocyte subsets and dendritic cell subsets in peripheral blood of primary hepatocellular carcinoma (HCC) patients after being transfused with autologous cytokine-induced killer cells (CIK) in patients, then to evaluate the clinical efficacy of the immune therapeutic strategy. METHODS: Peripheral blood mononuclear cells (PBMCs) from 13 patients with primary were collected using blood cell separator, and expanded in the fresh AIM-V medium in the presence of cytokine cocktail including interferon-gamma (IFN-gamma), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). The phenotypic patterns of CIK cells were longitudinally characterized by flow cytometry on day 0, 4, 7, 10,13 and 15 during the incubation period. PBMCs obtained from HCC patients before or after CIK cells transfusion into bodies to assay the changes of proportion of DC1 or DC2 in peripheral blood. RESULTS: After in vitro incubation for 14 or 15 days, a large of CD3(+)CD56(+) cells were produced from their progenitors and the percentages of CD3(+)CD8(+), CD3(+)CD56(+), CD25(+) cells significantly increased from 33.5% +/- 10.1%, 7.7% +/- 2.8%, and 12.3% +/- 4.5% at the beginning to 36.6% +/- 9.0% (P < 0.05), 18.9% +/- 6.9% (P < 0.01), and 16.4% +/- 5.9% (P < 0.05) at the day 15, respectively. In contrast, the percentages of CD3(+)CD4(+) and NK cells displayed no significant difference. The percentages of CD3(+), CD3(+)CD8(+) cells was held at a higher level during the whole incubation period, however those of the CD25(+), and CD3(+)CD56(+) cells began decreasing on day 7 and day 13, respectively. The proportion of type I of dendritic cells (DC1) and type II of dendritic cells (DC2) subsets increased from 0.59% +/- 0.23% and 0.26% +/- 0.12% before CIK cell transfusion to 0.85% +/- 0.27% and 0.43% +/- 0.20% (all P < 0.01) after CIK cell transfusion. The symptom of HCC patients receiving the CIK cell therapy was markedly ameliorated, and not side effect was seen in the treatment. CONCLUSION: Our results indicated that autologous CIK cells is able to boost the cellular immunological function in HCC patients, which probably provide a potent immune therapeutic strategy for HCC patients.

Adult↗
PubMed2003

[Epidemiologic features, clinical diagnosis and therapy of first cluster of patients with severe acute respiratory syndrome in Beijing area].

OBJECTIVE: Investigate the features of outbreak epidemic, clinical disease progression of the first SARS cases in Beijing and evaluate the efficacy of therapeutic regimen. METHODS: Twenty-nine patients (11 men and 18 women, 20 - 74 years old age range) were diagnosed with infectious SARS and admitted in our hospital from the March 5th to April 14th, 2003 in this study. The data of clinical presentation and disease progression of all the patients including index subject as the infectious SARS resource patient, her family infected members and 21 health care workers were abstracted. RESULTS: The first SARA outbreak in Beijing was characterized with the cluster feature of resource patient family members and health care providers. The incubation period ranged from 2 to 14 days. All the patients had a fever (temperature > 38 degrees C for over 24 hours) and other manifestations as reported before. Serial chest radiographs showed progressive pathologic air-space disease. Twenty patients showed the severe syndrome with various time ranged from 1 day to 14 days. Two patients died of progressive acute respiratory distress disease. The histologic analysis of one death patient showed diffuse alveolar damage in the two lungs. Twenty-six patients receiving the combined therapy including use of corticosteroid, antiviral ribavirin agents after the onset of symptoms and showed they had an acute self-limited disease course. The oldest patient (74 year old, male) received the healthy convalescent plasma infusion (50 ml) from recovered SARS subject and completely recovered within 21 days, having a shorter disease course. CONCLUSION: SARS is a kind of new self-limited and acute infectious disease. Early diagnosis, early isolation, early antiviral therapy for patients and efficient prevention for health care providers are urgently recommended. In particular, a combinational therapy of use of antiviral agents, preventive antibacterial antibiotics and pulsed dosage of corticosteroid can efficiently raise the clinical recovery rate and decrease mortality of SARS patients.

Adult↗
PubMed2003

[Identification of phenotype and Interferon-alpha-producing capability of circulating type II dendritic cells and its clinical implication in HBV-infected patients].

OBJECTIVE: To investigate the number, phenotype, and interferon-alpha (INF-alpha) of type II dendritic cells (DC2) in persons with hepatitis B and evaluate the role of DC2 subset in the immunopathogenesis of chronic HBV infection. METHODS: Peripheral blood was extracted from 103 hepatitis B (HB) virus-infected persons, including 11 cases of HB virus (HBV)-infected persons, 11 cases of acute HB, 81 cases of chronic HB, and 11 cases of asymptomatic HBV infection, and 25 healthy blood donors used as controls. Flow cytometry was used to calculate the number and the phenotype of circulating DC2. Ultraviolet-inactivated herpes simplex virus (HSV)-1 was added into the suspension of peripheral blood mononuclear cells (PBMCs) and then co-cultured for 24 hours to stimulate the production of INF-alpha by DC2 that was examined by ELISA assay. RESULTS: The number of DC2 in patients with chronic HB was 3.3 +/- 1.0 10(6)/L, significantly lower than that in the healthy controls (7.2 +/- 2.4 10(6)/L, P < 0.01). However, the number of DC2 was not significantly different between any other groups. The proportion of GS2 to PBMCs in the patients with chronic HB was 1.12 +/- 1.13 approximately 0.22 +/- 0.10, all significantly lower than that in the healthy controls (0.32% +/- 0.13%, P < 0.01). However, the proportion of GS2 to PBMCs was not significantly different between any other groups. The decrease of number of DC2 and that of proportion of DC2 to PBMCs in patients with chronic HB were related with the progress of disease. The INF-alpha concentration in the suspensions of PBMCs of different groups without stimulation by HSV-1 were low and there was no significant difference in INF-alpha concentration between different groups. The INF-alpha concentration in the suspension of PBMCs of healthy controls was 789 +/- 82 pg/ml, significantly higher than those of the patients with acute HB (161 +/- 36 pg/ml) and the patients with chronic HB (183 +/- 113 pg/ml, 147 +/- 39 pg/ml, and 156 +/- 39 pg/ml, all P < 0.05). However, there was no significant difference between the patient groups (all P > 0.05). CONCLUSION: The number and INF-alpha producing function of DC2, and the numbers of NK cells and CD8+ T cells in peripheral blood of patients with chronic HB decrease significantly, which results the deficiency of HBV-specific immune response.

Adolescent↗
PubMed2003

[Single nucleotide polymorphism loci of HIV-1 coreceptor CCR5 gene in Chinese Han people].

OBJECTIVE: To investigate the single nucleotide polymorphism(SNP) loci of HIV-1 coreceptor CCR5 gene in Chinese Han people. METHODS: The coding region of CCR5 was amplified using 2 pairs of primers and the PCR products of all 42 healthy subjects were sequenced by 4 different primers. The results of sequencing were analyzed by DNAstar in search of SNP loci. RESULTS: Six SNP loci were discovered in the coding region of CCR5, among them four SNPs, i.e. 184A-->G, 503G-->T, 688G-->A and 999G-->T, cause amino acids changes and two SNPs are nonsense mutations. One cytosine deletion at the 894nt results in frame shift mutation and prematured termination. 184A-->G, 503G-->T and 999G-->T were found in Chinese Han people for the first time. The allelic frequencies of mutant 184G, 503T and 999T alleles were 1.1%, 21.1% and 10.0% in healthy Hans, respectively. The population distribution of G503T markedly deviated from Hardy-Weinberg equilibrium. CONCLUSION: The SNP loci in the coding region of CCR5 in Chinese Han people has its own characteristics, which is not consistent with those of Japanese and obviously different from those of Caucasian and African.

Adult↗
PubMed2003

[Polymorphism of CXCR4 coding region of human immunodeficiency virus-1 in Chinese Han people].

OBJECTIVE: To study the polymorphism of human immunodeficiency virus (HIV)-1 coreceptor CXCR4 in Chinese Han ethnic group for AIDS prevention and treatment. METHODS: Totally 48 individuals were enrolled into the study. CXCR4 (cDNA No-AF147204) was cloned by PCR amplification using 2 pairs of primers, then sequenced using sequencing primers. The results of the same sequencing primers were analyzed by DNAstar software to find and identify single nucleotide polymorphism (SNP) sites. RESULTS: Totally 7 SNPs were found in the coding region of CXCR4, among them 3 were synonymous mutation (C-->T at loci 129, 426 and 968), 3 were missense mutation (C-->T at locus 38, A-->T at locus 90, and A-->C at locus 712) and 1 was stop mutation (C-->T at 106, which converted the codon for glutamic acid into stop codon). CONCLUSIONS: The polymorphism of CXCR4 coding region in Chinese Han is probably different from that of the other ethnic groups. Six of the 7 SNPs were discovered for the first time. Their influences on AIDS progression are worthy of studying.

Adult↗
PubMed2003

[Distribution of HIV-1 coreceptor CX3CR1 allelic polymorphisms in general population, HIV-1 high-risk group and HIV-1 carriers of Chinese indigenous Han and Uygur people].

OBJECTIVE: To investigate the V249I and T280M allelic polymorphisms of human immunodeficiency virus (HIV) coreceptor CX3CR1 in HIV-1 infected and uninfected population of Chinese indigenous Han and Uygur people and to probe the association between I249-M280 haplotype and HIV-1 susceptibility as well as AIDS progression. METHODS: Genomic DNA of 223 Uygur subjects and 316 Han subjects were purified from PBMC. I249 and M280 allelic frequencies were identified by polymerase chain reaction (PCR)/nest polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. All data were tested by chi(2) or u statistics analysis. RESULTS: Allelic frequencies of I249 and M280 were 16.1% and 13.3% in Uygur people, and 3.3% and 2.4% in Han people. No obvious difference existed between three groups of either ethnic group. However the allelic frequencies of HIV infected population were higher than those of general population, and those of general population higher than those of HIV-1 high-risk group. There was a strong linkage between I249 and M280 (P almost zero). CONCLUSIONS: I249 mutation was the sine qua non of M280 mutation, and most I249 alleles were accompanied by M280. The frequency of I249-M280 haplotype in Uygur population (13.3%) was adjacent to Caucasian people (15.8%), and that of I249-T280 haplotype (2.8%) was obviously lower than Caucasian people (12.5%); while both of them in Han people were much lower (0.9% and 2.4%). I249-M280 haplotype could accelerate AIDS progression according to Faure et al, while might be associated with HIV-1 susceptibility.

Alleles↗
PubMed2003

[Antiviral effect of human CTLs activated by HBsAg stimulated dendritic cells in vitro].

OBJECTIVE: To investigate the ability of human dendritic cells (DCs) inducing specific T lymphocyte response and inhibit the expression of HBeAg and HBsAg in 2.2.15 cell culture supernatant. METHODS: DCs were prepared from peripheral blood mononuclear cells induced with granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin 4. DCs was impulsed with pure HBsAg before DCs maturation and cocultured with self-blood T lymphocyte, while DCs without pure HBsAg stimulated group, T lymphocyte group and only T lymphocyte group were prepared as control group. The culture supernatant of 2.2.15 cell with stimulated T lymphocytes was collected on day 1, day 3, day 5 and day 7, respectively. The expressed levels of HBeAg and HBsAg were detected by ELISA method. RESULTS: DCs after antigen stimulation had a strong ability to present antigen and induce immune activation, DCs after loading with antigen in normal control and chronic hepatitis patients group had stronger stimulative ability for T lymphocytes proliferation than that of DCs without loading with antigen and only T lymphocyte group(P less than 0.01). The stimulating ability of DCs had a positive correlation to the dosage of loaded antigen; CTLs produced as a result of DCs stimulation had a specific inhibitive effect on the expression of HBeAg in 2.2.15 cell supernatant,but not on the expression of HbsAg. CONCLUSION: Human dendritic cells stimulated with HBsAg in vitro can efficiently present antigen and stimulate the production of specific CTLs to HBV, which can efficiently inhibit the expression of HBeAg in 2.2.15 cell supernatant- DC vaccine may become an antiviral therapy strategy for chronic hepatitis B virus infected patients in future.

Adolescent↗
PubMed2002

A novel apoptosis research method with imaging-combined flow cytometer and HITC or IR-125 staining.

The most commonly used methods for apoptotic research include terminal transferase-mediated dUTP nick end-labeling, annexin V testing of phosphatidylserine translocation from the inner leaflet to the outer plasma membrane by flow cytometry, DNA electrophoresis, and cell morphology. These methods provide apoptotic information from different aspects. To find a new way in apoptosis research and potential clinical application, we recently developed a novel method with an imaging-combined flow cytometer (IFC) and an innovative cell staining process by using 2-[7-(1,3-dihydro-1,3,3-trimethyl-2H-indol-2-ylidene)-1,3,5-heptatrienyl]-1,3,3-trimethyl-3H-indolium iodide (HITC) and 2-[7-[1,3-dihydro-1,1-dimethyl-3-(4-sulfobutyl)-2H-benz[e]indol-2-ylidene]-1,3,5-heptatrienyl]-1,1-dimethyl-3-(4-sulfobutyl)-1H-benz[e]indolium hydroxide, inner salt, sodium salt (IR-125). The IFC used in the research is a new generation of cytometry designed for simultaneous observations of cell populations and images. This is possible because the IFC is equipped with dual laser beams, one argon and one infrared. A promyelocytic leukemia cell line, HL-60, was used in the research. The cells were stained with our newly developed HITC or IR-125 staining method and a traditional nucleic acid dye, propidium iodide. The cells stained with HITC or IR-125 appeared completely dark in the IFC image window before washing. Phosphate buffered saline wash did not change the cell appearance. A wash with 50% methanol caused the cells to have a clear cell image with bright nuclei on the IFC. To obtain apoptotic cells, we treated the HL-60 cells with 0.15 microM of camptothecin (CAM), a topoisomerase I inhibitor and experimental apoptosis inducer, for 4 h. The control showed larger round cells with bright nuclei and one to three dark nucleoli. The CAM-induced apoptotic cells were smaller, with fragmented and condensed nuclei on the IFC. These appearances were identical to the cell morphology of with light and electron microscopy. We used other methods including FACScan and DNA electrophoresis to confirm the apoptotic changes after CAM treatment and compared them with the IFC method. In addition, we found that the novel method with the IFC and HITC or IR-125 staining can show not only cell apoptotic changes but also peripheral blood cell populations and images simultaneously. This study suggests many potential applications of the IFC and this novel staining method in other cellular biological researches and clinical assays.

Apoptosis↗
PubMed2002

Immature granulocyte count after liver transplantation.

We evaluated the significance of immature granulocyte (IG) count during the clinical course after liver transplantation. We counted IG using the flow cytometric method with CD16, CD11b, and CD45 antibodies. Samples were obtained from 31 patients in the Department of Transplantation and Immunology, and we determined (i) the distribution of IG peak value, (ii) the distribution of IG peak time-points, (iii) the clinical background of patients with high IG, and (iv) the clinical course of high IG cases. We observed the appearance of IG (100/microl or higher) in the majority of the patients (23 out of 31 patients; 74.2%). The IG peak was detected on the 19th day after transplantation. We observed serious complications, such as melena, rejection, or severe infection, in high IG (500/microl or higher) cases. We observed instances of inflammation with low C-reactive protein (CRP) value in the presence of IG. We believe that IG is a useful marker to monitor inflammation.

Biomarkers↗