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1.
Article in English | WPRIM | ID: wpr-921355

ABSTRACT

Objective@#The coronavirus disease 2019 (COVID-19) pandemic continues to present a major challenge to public health. Vaccine development requires an understanding of the kinetics of neutralizing antibody (NAb) responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).@*Methods@#In total, 605 serum samples from 125 COVID-19 patients (from January 1 to March 14, 2020) varying in age, sex, severity of symptoms, and presence of underlying diseases were collected, and antibody titers were measured using a micro-neutralization assay with wild-type SARS-CoV-2.@*Results@#NAbs were detectable approximately 10 days post-onset (dpo) of symptoms and peaked at approximately 20 dpo. The NAb levels were slightly higher in young males and severe cases, while no significant difference was observed for the other classifications. In follow-up cases, the NAb titer had increased or stabilized in 18 cases, whereas it had decreased in 26 cases, and in one case NAbs were undetectable at the end of our observation. Although a decreasing trend in NAb titer was observed in many cases, the NAb level was generally still protective.@*Conclusion@#We demonstrated that NAb levels vary among all categories of COVID-19 patients. Long-term studies are needed to determine the longevity and protective efficiency of NAbs induced by SARS-CoV-2.


Subject(s)
Adult , Aged , Aged, 80 and over , Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , COVID-19/immunology , Female , Humans , Kinetics , Male , Middle Aged , Neutralization Tests , SARS-CoV-2
2.
Article in Chinese | WPRIM | ID: wpr-698337

ABSTRACT

BACKGROUND: Retrorsine (RS) is a chemical agent for the long-term inhibition of mature liver cell division and proliferation. OBJECTIVE: To establish a rat model of liver injury by combined use of RS and 1/3 partial hepatectomy, to observe the proliferation of liver cells and oval cells in rats after liver injury, and to discuss the relationship between liver regeneration and mature liver cells and oval cells after liver injury. METHODS: Thirty male Sprague-Dawely rats were randomized into two groups (n=15 per group): RS group and control group. Rats in the RS group were subjected to intraperitoneal injection of RS, 30 mg/kg, twice in total, with 2 weeks in between; and rats in the control group were injected physiological saline instead of RS. Four weeks after the last injection, the 1/3 partial hepatectomy was performed in two groups of rats. Liver pathological and morphological changes as well as cell proliferation were observed, and CK19 and C-kit immunohistochemical detections of the rat liver in the two groups were conducted at different time points after operation. RESULTS AND CONCLUSION: At 20 days after operation, the body mass of the RS group rats was still lower than the baseline, and the liver increase was obviously less than that in the control group; there was cell body swelling shown by hematoxylin-eosin staining, loose cytoplasm, extensive vacuoles degeneration of liver cells, and clustered or scattered oval cells around the portal area of small bile duct and in the hepatic lobule. However, in the control group, the body mass was close to the baseline, liver damage was lighter than that in the RS group, a large number of mature liver cells proliferated under BrdU Immunofluorescence at 20 days after operation; liver oval cells proliferated and distributed in the liver cell line at 14 days after operation, with morphology and immunohistochemical markers consistent with oval cells in the RS group. These findings indicate that the rat model of acute liver injury is successfully established by combining retrorsine with 1/3 partial hepatectomy, liver poisoning and the proliferation of liver stem cells and mature liver cells after poisoning can be seen in the experiment, which firmly confirm that liver cell renewal and regeneration after injury is accredited to the combined action of liver stem cells in liver basin and mature liver cells.

3.
Journal of Preventive Medicine ; (12): 757-761, 2017.
Article in Chinese | WPRIM | ID: wpr-792641

ABSTRACT

Objective To investigate the genetic toxicity of methyl tert-butyl ether (MTBE) to lymphocytes and to provide reference for establishing the occupational population exposure limit of MTBE. Methods The human B lymphocytes in the logarithmic growth phase were respectively exposed to MTBE at concentrations of 0, 1.25, 2.5, 5, 10 and 12.5 μmol/L. The percentage of tail DNA and Olive tail moment of human B lymphocytes were evaluated after 24 h exposure by using comet assay. Apoptosis was tested by Annexin V-FITC/PI double-staining flow cytometry. The contents of MDA and 8-OHdG, as well as GSH-Px activity were measured by ELISA kit. Sixty workers from a petrochemical factory in Zhejiang were selected as the occupational exposure population, and 55 non-occupational exposure workers were selected as the control population. And 5 mL heparin anticoagulant peripheral blood was collected, and the number of micronucleus of peripheral blood lymphocytes was measured by micronucleus test, and the percentages of tail DNA as well as Olive tail moment of peripheral blood lymphocytes were measured by comet assay. The contents of MDA and 8-OhdG and GSH-Px activity in peripheral blood plasma were measured by ELISA kit. Results After 24 h exposure, the percentage of tail DNA and Olive tail moment in human B lymphocytes at concentrations of 10-12.5 μmol/L were significantly higher than those in the control group (P<0.05), and the percentage of early apoptosis cells was significantly increased at concentrations of 10-12.5 μmol/L (P <0.01) . The results of population-based study indicated no statistically significant difference in micronucleus positive rate, and the contents of MDA and 8-OhdG, and GSH-Px activity between the exposure group and the control group, but the Olive tail moment was significantly higher in the exposure group compared with the control group (P=0.000) . Conclusion The results of vitro study showed that exposure to 10-12.5 μmol/L MTBE could cause genetic toxicity to human B lymphocytes. Olive tail moment of peripheral blood lymphocytes of occupational exposure workers was significantly higher than that of non-occupational exposure group.

4.
Article in Chinese | WPRIM | ID: wpr-318104

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the death causes of 345 cases with HIV/AIDS in Guangdong area.</p><p><b>METHODS</b>The situations of 345 hospitalized death cases with HIV/AIDS were conducted by retrospective analysis.</p><p><b>RESULTS</b>(1)There were total 3406 hospitalized cases with HIV/AIDS in a hospital from January 2001 to December 2011 and 345 cases died, the fatality rate was 10. 13%. Since 2005 the introduction of free anti-viral treatment, the fatality rate of HIV/AIDS declined. The fatality rate of the patients whose CD4+ T lymphocyte counts <200 cells/microl was 14.61% (299/2046) and it was significantly higher than that of patients whose CD4 T lymphocyte counts >or=200 cells/microl (P <0.01). (2) 99.42% of the death cases had more than one kind of opportunistic infections (OI) and there were 924 cases of OI totally. 84. 64% of OI related to the death directly. Fungal infection was the most common in OI, followed by bacterial infection. Most OI occurred in the lungs, mouth, other systemic disseminated diseases, gastrointestine, central nerver system, septicemia, skin. The AIDS defining opportunistic infections such as several pneumonia, disseminated penicilliosis marneffei and CNS infections accounted for 29.65%. Other factors that caused HIV/AIDS death included opportunistic tumors, HIV related disease and non AIDS-related disease accounted for 15.36%. No accepted effective highly active antiretroviral therapy (HARRT) also constituted factors of death. Among cases which accepted HARRT treatment, only 6.96% had the period of treatment over three months.</p><p><b>CONCLUSION</b>The fatality rate of end-stage AIDS patients was high and the opportunistic infections was the most important cause of death. Early diagnosis and treatment for opportunistic infections, timely effective HARRT were the key to improve the quality of life of AIDS patients.</p>


Subject(s)
Acquired Immunodeficiency Syndrome , Drug Therapy , Allergy and Immunology , Microbiology , Mortality , Adolescent , Adult , CD4 Lymphocyte Count , Methods , Cause of Death , Child , Child, Preschool , China , Epidemiology , Female , HIV Infections , Drug Therapy , Allergy and Immunology , Microbiology , Mortality , Humans , Infant , Infant, Newborn , Male , Middle Aged , Retrospective Studies , Young Adult
5.
Chinese Journal of Hepatology ; (12): 829-833, 2013.
Article in Chinese | WPRIM | ID: wpr-277988

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the potential differences in response to pegylated interferon (Peg-IFN) plus ribavirin (RBV) combination therapy in patients with hepatitis C virus (HCV) mono-infection and human immunodeficiency virus (HIV)/HCV co-infection.</p><p><b>METHODS</b>Seventy HIV/HCV patients and sixty HCV patients, were administered a 48-week course of Peg-IFN + RBV. The HCV load was tested by the COBAS automatic viral load analysis system (lower limit of quantification = 15 IU/ml) at treatment weeks 0 (baseline), 4, 12, 24, and 48 and at week 24 after drug withdrawal. The patients were also genotyped by sequencing for the host-encoded interleukin (IL)-28B single nucleotide polymorphisms (SNPs) related to HCV Peg-IFN + RBV therapy outcome: rs8099917, rs12979860 and rs12980275. In addition, the HCV-encoded NS5B gene region was genotyped by nested-PCR and sequencing followed by BLAST searching of the Los Alamos National Laboratory HCV database. The significance of between-group differences in response to therapy and roles of SNPs were evaluated by statistical analyses.</p><p><b>RESULTS</b>The ratio of sustained virological response (SVR) was significantly lower in the HIV/HCV co-infected patients than the HCV mono-infected patients (32.9% vs. 71.7%; P less than 0.001). While the HIV/HCV co-infected patients did not show a significant difference in SVR ratio achieved between individuals infected with the HCV-1 genotype and the non-HCV-1 genotype (30.8% vs. 33.3%; P = 1.000), the HCV mono-infected patients did (86.1% vs. non 50.0%, P = 0.002). Moreover, the SVR ratio was higher in the HCV-1 genotype HCV mono-infected patients than in the HIV/HCV-1 genotype co-infected patents (30.8% vs. 86.1%; P less than 0.001). The different IL-28B genotypes were not significantly correlated to the PEG-IFN+RBV therapy response of either HCV mono-infected patients or HIV/HCV co-infected patients (P more than 0.05).</p><p><b>CONCLUSION</b>HCV mono-infected patients respond better to Peg-IFN + RBV therapy than HIV/HCV co-infected patients. The HCV-1 genotype may promote this therapy response in HCV mono-infected patients, but the IL-28B genotypes appear to play no significant role.</p>


Subject(s)
Adult , Antiviral Agents , Therapeutic Uses , Coinfection , Drug Therapy , Drug Therapy, Combination , Female , Genotype , HIV Infections , Drug Therapy , Virology , Hepacivirus , Hepatitis C, Chronic , Drug Therapy , Virology , Humans , Interferon-alpha , Therapeutic Uses , Interleukins , Genetics , Male , Polyethylene Glycols , Ribavirin , Therapeutic Uses , Treatment Outcome
6.
Chinese Medical Journal ; (24): 823-827, 2012.
Article in English | WPRIM | ID: wpr-262519

ABSTRACT

<p><b>BACKGROUND</b>Penicillium marneffei (P. marneffei) is an emerging pathogenic fungus that can cause invasive mycosis in patients with AIDS. The epidemiological features of P. marneffei infection in AIDS patients in Guangdong province remain unclear so far. This study aimed to investigate the genetic diversity within a population of 163 P. marneffei isolates obtained from AIDS patients and search for the dominant clinical strains in Guangdong province.</p><p><b>METHODS</b>One hundred and sixty-three P. marneffei isolates obtained from AIDS patients in Guangdong province during January 2004 and December 2009 were studied by randomly amplified polymorphic DNA (RAPD) using two random primers (H2 and H22). The degree of similarity between samples was calculated through similarity coefficients from RAPD fragment data and the dendrogram was assessed using the unweighted pair group method with arithmetic mean (UPGMA).</p><p><b>RESULTS</b>Two primers showed a high degree of discrimination and good stability. Primer H2 yielded eight different patterns (H2-1 to H2-8) among 163 isolates with the discriminatory power being 0.413. Primer H22 identified seven types (H22-1 to H22-7) among 163 isolates with the discriminatory power being 0.467. Genetic similarity coefficients based on RAPD data among 163 P. marneffei isolates ranged from 0.681 to 0.957, 61.96% of which were no less than 0.83. The discriminatory power of the two primers was 0.524. One hundred and sixty-three P. marneffei isolates were clustered into nine distinct groups (groups I to IX) at the similarity coefficient value of 0.83 and group I was the most common, including 101 strains (61.96%).</p><p><b>CONCLUSION</b>The RAPD analyses could provide important information as to the degree of genetic diversity and the relationship among clinical P. marneffei isolates, revealing genetic polymorphism and dominant strains.</p>


Subject(s)
Acquired Immunodeficiency Syndrome , Microbiology , Genetic Variation , Genetics , Humans , Penicillium , Classification , Genetics , Random Amplified Polymorphic DNA Technique , Methods
7.
Chinese Journal of Hepatology ; (12): 729-933, 2011.
Article in Chinese | WPRIM | ID: wpr-239339

ABSTRACT

<p><b>OBJECTIVE</b>To explore the transmission routes, genotypes/subtypes distribution and genetic character of HCV in HIV/HCV co-infected and HCV mono-infected individuals in Guangdong Province.</p><p><b>METHODS</b>Reverse transcription (RT) nested PCR was performed to amplify the HCV NS5B gene region from 95 HIV/HCV co-infected and 99 HCV mono-infected individuals lived in Guangdong province. The PCR products were then sequenced for HCV subtyping. Genetic analysis was done by MEGA4 software.</p><p><b>RESULTS</b>(1) HIV/HCV co-infected individuals infected HCV mostly through injection drug use (IDU, 78.9%), the HCV subtypes were identified as 6a (53.7%), 3a (17.9%), 1b (15.8%), 3b (11.6%) and 1a (1.0%) respectively, the genetic distance within subtype 1b was longer than those within other subtypes, the predominant HCV subtype in HIV/HCV co-infected individuals infected through IDU was 6a (60.0%). (2) HCV mono-infected individuals infected HCV mostly through blood or blood products transfusions (80.8%), the HCV subtypes were identified as 1b (67.7%), 6a (17.2%), 3a (6.1%), 2a (5.0%), 3b (2.0%), 4a (1.0%) and 5a (1.0%) respectively, the genetic distance within subtype 1b was also longer than those within other subtypes, the predominant HCV subtype in HCV mono-infected individuals infected through blood or blood products transfusions was 1b (76.2%).</p><p><b>CONCLUSION</b>The diversity of HCV subtypes in HIV/HCV co-infected and HCV mono-infected individuals in Guangdong Province was high, both the major transmission route and HCV subtype between HIV/HCV co-infected individuals and HCV mono-infected individuals were different.</p>


Subject(s)
Adolescent , Adult , Aged , Asians , China , Epidemiology , Coinfection , Virology , Female , Genotype , HIV , HIV Infections , Epidemiology , Virology , Hepacivirus , Classification , Genetics , Hepatitis C , Epidemiology , Virology , Humans , Male , Middle Aged , Phylogeny , Young Adult
8.
Article in Chinese | WPRIM | ID: wpr-313465

ABSTRACT

<p><b>OBJECTIVE</b>To discuss the origin of occupational stress among petrochemical industry workers and to access the main occupational stressors that impact job satisfaction and mental health of petrochemical industry workers.</p><p><b>METHODS</b>A survey on occupational stressor was carried out by Occupational Stress Indicator (OSI) in 532 petrochemical industry workers (345 chemical and 187 logistic workers).</p><p><b>RESULTS</b>The environment in workplace of chemical group was worse than that of contrast. The chemical workers had less control over job and they experienced more hazards, monotonous as well as role stressors than the logistic group. The scores of job satisfaction and mental health of chemical group (36.867 +/- 0.656, 43.734 +/- 0.542, respectively) were higher than that of contrast (40.321 +/- 0.901, 46.714 +/- 0.745, respectively) (P < 0.05).</p><p><b>CONCLUSION</b>The occupational stressors exist in chemical workers which affect chemical workers' job satisfaction and mental health with different levels.</p>


Subject(s)
Adult , Analysis of Variance , Burnout, Professional , Chemical Industry , Female , Humans , Job Satisfaction , Male , Middle Aged , Petroleum , Regression Analysis , Young Adult
9.
Article in Chinese | WPRIM | ID: wpr-332441

ABSTRACT

<p><b>OBJECTIVE</b>To explore the relationship between psychological distress and T lymphocyte counts in HIV/AIDS patients.</p><p><b>METHODS</b>A total of 102 HIV/AIDS patients were measured by symptom check list (SCL-90), self-rating depressive scale (SDS) and self-rating anxiety scale (SAS). Patients were divided into 2 groups based on CD4+ T lymphocyte counts < 0.2 x 10(9)/L (group A) and > or = 0.2 x 10(9)/L(group B).</p><p><b>RESULTS</b>77 cases (75.49%) had psychological problems, including depression, relationship problems, psychosis, force etc. The prevalence of depression and anxiety were 67.65% (69/102) and 43.13% (44/102) respectively. The symptom of depression and anxiety of patients in group A were severer than those in group B (P < 0.05). The CD4+ T lymphocyte counts were significantly negatively correlated with the total score, depression score, paranoid score and psychosis score of SCL-90 (all P <0.05).</p><p><b>CONCLUSION</b>Most of the HIV/AIDS patients were in an obviously abnormal psychological status. The psychological distress symptom of HIV/AIDS patients might had negative effects on the number of CD4+ T lymphocyte.</p>


Subject(s)
Acquired Immunodeficiency Syndrome , Allergy and Immunology , Psychology , Anxiety , CD4 Lymphocyte Count , Depression , Female , HIV Infections , Allergy and Immunology , Psychology , Humans , Male , T-Lymphocytes , Allergy and Immunology
10.
Article in Chinese | WPRIM | ID: wpr-332439

ABSTRACT

<p><b>OBJECTIVE</b>To test the infeciton efficiency of recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and to abserve its biological characteristics.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMC) were separated from healthy blood donor and they were infected by Ad-HSP70-HBsAg on the first day of isolation. DCs were induced in medium with cytokines IL-4, GM-CSF and TNF-alpha in vitro. The biological characteristics of DC induced were analyzed by inverted fluorecent microscope, RT-PCR, flow cytometer (FACS), and mixed lymphocyte reaction (MLR).</p><p><b>RESULTS</b>The traced gene-GFP were abserved in DCs by inverted fluorecent microscope and HSP70-HBsAg gene mRNA expression was detected by RT-PCR after the Ad-HSP70-HBsAg infection. FACS analysis shown that the expression of CD1a, CD80, CD86 and HLA-DR on surfece of two groups of DCs were similar. MLR showed that there are not a statitic difference of stimulated index (SI) between two groups.</p><p><b>CONCLUSION</b>Results indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics.</p>


Subject(s)
Adenoviridae , Genetics , Physiology , Cells, Cultured , Cytokines , Genetics , Allergy and Immunology , Dendritic Cells , Allergy and Immunology , Virology , Genetic Vectors , Genetics , HSP70 Heat-Shock Proteins , Genetics , Allergy and Immunology , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Humans , Lymphocyte Culture Test, Mixed , Recombinant Fusion Proteins , Genetics , Allergy and Immunology
11.
Article in Chinese | WPRIM | ID: wpr-325125

ABSTRACT

<p><b>OBJECTIVE</b>To survey the dose of glucocorticosteroids administered in patients with severe acute respiratory syndrome (SARS) and assess the effect of glucocorticosteroid doses in improving the patients' lung function.</p><p><b>METHODS</b>A retrospective analysis was conducted among 225 SARS patients treated in our in 2003. Oxygenation index was used as the effectness index, and the criteria for effectiveness was defiend as increase of the value of OI by 20% or above.</p><p><b>RESULTS</b>Glococoticostecoids were used in 59.56% of the SARS cases. The average value of OI before intravenous use of glucocorticosteroids was 237.08 mmHg, and that after the administration was 335.08 mmHg. The glucocorticosteroid doses that produce better effects were 1-3 mg/kg and 160-240 mg daily, with the total accumulative dose of 1000-2000 mg. The optimal duration of glucocorticosteroid use was 8-14 days.</p><p><b>CONCLUSIONS</b>For SARS treatment, Glucocorticosteroids can effectively ameliorate the SARS patients' lung symptoms and improve the lung function. The appropriate daily dose of glucocorticosteroids is 1-3 mg/kg or 160-240 mg/d for a duration of 8-14 d; the accumulative dose should be controlled around 1500 mg.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Glucocorticoids , Humans , Male , Methylprednisolone , Middle Aged , Respiratory Function Tests , Respiratory Insufficiency , Retrospective Studies , Severe Acute Respiratory Syndrome , Drug Therapy , Young Adult
12.
Article in Chinese | WPRIM | ID: wpr-254119

ABSTRACT

<p><b>OBJECTIVE</b>To construct a recombinant adenoviral vector carrying HBcAg-HSP70 chimeric gene by homologous recombination in bacteria and to detect its expression in vitro.</p><p><b>METHODS</b>Heat shock protein 70 gene from Mycobacterium tuberculosis were amplified by PCR and were cloned to adenoviral shuttle plasmid pAdTrack-CMV-HBsAg. Then the resultant pAdTrack-CMV-HBsAg-HSP70 was cotransfected into BJ5183 bacteria with the plasmid pAdeasy-1. The adenoviral plasmid carrying HBsAg-HSP70 gene (pAd-HBsAg-HSP70) was generated with homologous recombination in bacteria and the adenoviruses were produced in 293 cells. Several kinds of mammal cells (293 cells and Vero cells) were infected with adenoviruses and the expression of HBsAg-HSP70 was detected by RT-PCR and ELISA in vitro.</p><p><b>RESULTS</b>The adenoviral plasmids pAd-HBsAg-HSP70 were obtained by selection for kanamycin resistance and confirmed by restriction endonuclease Pac analyses. The recombinant adenoviruses Ad-HBsAg-HSP70 were packaged successfully in 293 cells. The titer of Ad-HBsAg-HSP70 was up to 2 x 10(12) pfu/L after the second passage of proliferation in 293 cells. HBsAg and HSP70 were expressed efficiently in mammal cells after infection.</p><p><b>CONCLUSION</b>The recombinant adenoviruses expressing HBsAg and HSP70 were constructed successfully which can be used further in study of gene therapy for HBV.</p>


Subject(s)
Adenoviridae , Genetics , Animals , Cell Line , Chlorocebus aethiops , Defective Viruses , Genetics , Enzyme-Linked Immunosorbent Assay , Green Fluorescent Proteins , Genetics , Metabolism , HSP70 Heat-Shock Proteins , Genetics , Metabolism , Hepatitis B Surface Antigens , Genetics , Metabolism , Humans , Microscopy, Fluorescence , Recombinant Fusion Proteins , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transfection , Vero Cells , Virus Replication , Genetics
13.
Article in Chinese | WPRIM | ID: wpr-680045

ABSTRACT

Objective To investigate human immunodeficiency virus type 1 (HIV 1)specific CD8'T cell responses and related human leukocyte antigenⅠ(HLA-I)restriction sites.Methods Four Epstein-Barr virus (EBV)strains-transformed B-lymphoblastoid cell lines (BLCL)from periph eral blood monocytes (PBMC)of 4 healthy individuals were established.Using synthesized peptides in HIV-1 Gag,Vif and Nef regions as epitopes and using BLCL as antigen presenting cells (APC), the frequency of interferon-7(IFN-?)secreting cells in CD8 T cells from a HIV-infected long term nonprogressor by enzyme-linked immunospot (ELISPOT)assay in vitro were assessed.Moreover, matching analysis of HI.A-Ⅰrestriction sites were performed.Results HIV-1 epitopes were presen ted restrictively by HLA-Ⅰmolecules in specific CD8 T cell responses,and this antigen presenting had high specificity.HLA-Ⅰrestriction sitesA03,A26,C04andC08 contributed to presentepitopes in HIV-1 C-ag region,A03 and C08 to HIV-1 Nef as well.Conclusion HLA-Ⅰrestriction sites may include A03,A26,C04 and C08 in HIV-1 Gag region,and A03,C08 in Nef region.

14.
Article in Chinese | WPRIM | ID: wpr-248833

ABSTRACT

<p><b>OBJECTIVE</b>To explore whether hepatitis B virus (HBV) S gene-modified dendritic cells (DCs) might induce a specific cytotoxic T lymphocyte (CTL) response.</p><p><b>METHODS</b>The recombinant adenoviruses carrying HBsAg genes were prepared and used to transfect DCs generated from cord blood. The efficacy of transfection was observed through the expression of enhanced green fluorescent protein (EGFP) in DCs and the expression of HBsAg was detected by ELISA. HBV S gene-modified DCs were co-cultured with T cells from cord blood and T cells stimulating activities were detected using mixed lymphocyte reaction (MLR). The CTL assay was carried out to assess the ability of CTL lines to lyse target cells of HepG(2)22.1.5 by measuring lactate dehydrogenase (LDH) release.</p><p><b>RESULTS</b>The results showed that HBV S genes were expressed in DCs with high efficacy by recombinant adenoviral vector. DCs had a normal shape after transfection. The result of MLR showed that HBV S gene-modified DCs could effectively stimulate naive T cells to proliferate. The induced specific CTL lines could lyse target cells of HepG(2)22.1.5.</p><p><b>CONCLUSIONS</b>HBV S gene-modified DCs enhanced the function to induce a specific CTL effect, showing its promise for developing anti-viral vaccine in future.</p>


Subject(s)
Cell Line , Cells, Cultured , Cytotoxicity, Immunologic , Dendritic Cells , Allergy and Immunology , Virology , Hepatitis B , Allergy and Immunology , Virology , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Hepatitis B virus , Genetics , Allergy and Immunology , Humans , Lymphocyte Culture Test, Mixed , T-Lymphocytes, Cytotoxic , Allergy and Immunology
15.
Article in Chinese | WPRIM | ID: wpr-248828

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinical characteristics of the patients with dengue fever (DF) seen from 2002 to 2006 in Guangzhou in order to prevent and treat dengue fever better.</p><p><b>METHODS</b>Clinical data from 1342 inpatients with DF seen from 2002 to 2006 were retrospectively analyzed. The dengue virus was isolated by C6/36 cell culture and genotyped by reverse transcriptase-polymerase chain reaction and gene sequence analysis.</p><p><b>RESULTS</b>The average age of the patients was 34.4 years, without sex difference in distribution. Most of the patients had obvious toxemic symptoms including fever (100 percent), headache (85.9 percent), myalgia (64.5 percent), bone soreness (46.6 percent) and skin rash (65.9 percent). Leukopenia, thrombocytopenia, elevated alanine aminotransferase, elevated aspartate aminotransferase and hypokalemia were found in 66.0 percent, 61.3 percent, 69.0 percent , 85.7 percent and 28.4 percent of patients, respectively. DF-IgM could be detected in 90 percent of patients. The virus was identified as dengue virus type-I.</p><p><b>CONCLUSIONS</b>The epidemic of DF was caused by dengue virus- I from 2002 to 2006 in Guangzhou. Most of the patients had classic DF clinical manifestation with high percentage of hepatic injury. Few patients progressed to dengue hemorrhagic fever.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Antibodies, Viral , Blood , Child , Child, Preschool , China , Epidemiology , Dengue , Diagnosis , Epidemiology , Allergy and Immunology , Dengue Virus , Genetics , Female , Humans , Infant , Male , Middle Aged , Retrospective Studies , Young Adult
16.
Article in Chinese | WPRIM | ID: wpr-248799

ABSTRACT

<p><b>OBJECTIVE</b>To investigate immunodominance in CD8+ T cell responses to human immunodeficiency virus type 1 (HIV-1) epitopes.</p><p><b>METHODS</b>Frequency of Interferon-gamma (IFN-gamma) secreting cells and the proliferation percentage of CD8+ T cells in PBMC from an HIV-1-infected long term nonprogressor (LTNP) were assessed after stimulation with either the 34 pools of 701 overlapping peptides covering the regions of HIV-1 Env, Pol, Gag, Vif, Nef, Tat or some single peptides, by using various assays including enzyme-linked immunospot (ELISPOT) and CFSE Carboxy-fluorescein diacetate, succinimidyl ester (CFSE) labeling and flow cytometry.</p><p><b>RESULTS</b>HIV-1 Gag peptides induced the highest frequency of IFN-gamma secreting cells, followed by Nef, Tat, and Vif. Meanwhile, Env and Pol failed to induce significant responses. In the IFN-gamma ELISPOT assay, stimulation with single peptide and the corresponsive peptide pool generated analogous results. In addition, the frequencies of IFN-gamma secreting cells and the proliferation percentage of CD8+ T cells detected-ELISPOT and CFSE labeling and flow cytometry were proportional, when single peptides were used for stimulation.</p><p><b>CONCLUSION</b>CD8+ T cells can respond to some specific HIV-1 epitopes and induce immunodominant responses. As a complimentary approach to the standard of ELISPOT assay, We recommend a novel CFSE labeling and flow cytometry assay for the examination of immunodominance in studies of HIV-1 specific proliferation percentage of CD8+ T cell responses.</p>


Subject(s)
Adult , CD8-Positive T-Lymphocytes , Allergy and Immunology , Metabolism , Enzyme-Linked Immunosorbent Assay , Methods , Female , HIV Infections , Allergy and Immunology , Virology , HIV-1 , Allergy and Immunology , Humans , Immunodominant Epitopes , Allergy and Immunology , Pharmacology , Interferon-gamma , Metabolism
17.
Article in Chinese | WPRIM | ID: wpr-248781

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effect of diammonium glycyrrhizinate on the antiviral therapy with adefovir dipivoxil (ADV) in patients with HBeAg-positive chronic hepatitis B.</p><p><b>METHODS</b>Patients with HBeAg-positive chronic hepatitis B enrolled in this study were randomized to receive either ADV 10 mg/d fir 48 weeks or placebo for 24 weeks followed by ADV 10 mg/d for 24 weeks. Antiviral activities of diammonium glycyrrhizinate administered during the trial were studied with respect to virological and serologic response, and ALT normalization.</p><p><b>RESULTS</b>Twenty-one of 142 patients in ADV group vs. 11 of 68 patients in placebo group were treated with diammonium glycyrrhizinate. There was no significant difference in virological, serological and biochemical responses between patients with or without diammonium glycyrrhizinate in both therapy groups. During double-blind period, virological response was significantly worse in patients only receiving diammonium glycyrrhizinate than those combined with ADV.</p><p><b>CONCLUSION</b>Diammonium glycyrrhizinate had no antiviral activity and exerts no influence on the efficacy of ADV treatment in patients with HBeAg-positive chronic hepatitis B.</p>


Subject(s)
Adenine , Therapeutic Uses , Adult , Antiviral Agents , Therapeutic Uses , Double-Blind Method , Glycyrrhizic Acid , Therapeutic Uses , Hepatitis B e Antigens , Blood , Hepatitis B virus , Allergy and Immunology , Hepatitis B, Chronic , Blood , Drug Therapy , Virology , Humans , Male , Organophosphonates , Therapeutic Uses , Treatment Outcome
18.
Article in Chinese | WPRIM | ID: wpr-248777

ABSTRACT

<p><b>OBJECTIVE</b>To determine the relationship between the response to adefovir dipivoxil (ADV) treatment in patients with HBV genotypes B and C of HBeAg positive chronic hepatitis B.</p><p><b>METHODS</b>This clinical trial was a randomized, double-blind, placebo-controlled, multicenter study. A total of 226 eligible patients with HBeAg positive chronic hepatitis B were randomized (in a ratio of 2:1) receiving ADV 10 mg/d for 48 weeks (ADV+ADV group) or placebo for 24 weeks followed by ADV 10 mg/d for 24 weeks (PLB+ADV group). The primary efficacy was virologic response. The genotypes of HBV were determined by PCR-restricted fragment length polymorphism (RFLP) method using serum samples before therapy. rtN236T and rtA181V mutations were confirmed by sequencing.</p><p><b>RESULTS</b>In this study, HBV genotype C was 66.7%, genotype B was 25.2%. Genotype B was more common in Guangzhou. Patients with genotype B were much younger than those with the genotype C. Patients with genotype B previously received less anti-HBV therapy. There were no significant difference in virological response (including mean reduction in HBV DNA level from baseline, serum HBV DNA load after treatment and HBV DNA undetectable rate) and serological response (the rate of HBeAg loss and HBeAg seroconversion) between patients infected with genotypes B and C in both treatment arms.</p><p><b>CONCLUSION</b>There were no significant difference in virological and serological response to ADV therapy between patients infected with HBV genotype B and C.</p>


Subject(s)
Adenine , Therapeutic Uses , Adolescent , Adult , Antiviral Agents , Therapeutic Uses , DNA, Viral , Blood , Genetics , Double-Blind Method , Female , Genotype , Hepatitis B Antibodies , Blood , Hepatitis B e Antigens , Allergy and Immunology , Hepatitis B virus , Genetics , Allergy and Immunology , Hepatitis B, Chronic , Blood , Drug Therapy , Virology , Humans , Male , Middle Aged , Organophosphonates , Therapeutic Uses , Treatment Outcome , Young Adult
19.
Article in Chinese | WPRIM | ID: wpr-268118

ABSTRACT

<p><b>OBJECTIVE</b>To detect CCL20 and CXCR4 expressions in epidermis infected with condyloma acuminatum (CA) and normal epidermis and investigate the effect of their expressions on Langerhans cells in CA epidermis.</p><p><b>METHODS</b>Gene expression of CCL20 and CXCR4 in 3 epidermal CA lesions and in 3 normal epidermis specimens were detected using Affymetrix oligonucleotide microarrays HG-U 133A 2.0, and the protein levels of CCL20 and CXCR4 in these specimens were measured by Western blotting.</p><p><b>RESULTS</b>Microarray analysis revealed markedly down-regulated mRNA expressions of CCL20 and CXCR4 in the 3 epidermal CA lesions as compared with those in the normal specimens. Western blot analysis showed that the protein expressions of CCL20 and CXCR4 in the CA lesions were significantly lower than those in normal epidermis.</p><p><b>CONCLUSION</b>The protein and mRNA expressions of CCL20 and CXCR4 are markedly down-regulated in epidermal CA lesions, which may contribute to decreased number and backflow disturbance of Langerhans cells in these lesions.</p>


Subject(s)
Adult , Blotting, Western , Chemokine CCL20 , Genetics , Metabolism , Condylomata Acuminata , Genetics , Metabolism , Down-Regulation , Epidermis , Metabolism , Pathology , Gene Expression Regulation , Humans , Oligonucleotide Array Sequence Analysis , RNA, Messenger , Genetics , Metabolism , Receptors, CXCR4 , Genetics , Metabolism , Young Adult
20.
Article in Chinese | WPRIM | ID: wpr-333050

ABSTRACT

<p><b>OBJECTIVE</b>To study HBV preS2/S gene expression effects in mammalian cells transferred with recombinant adenoviral vector.</p><p><b>METHODS</b>The replication-deficient recombinant adenoviral vector (Ad-HBs) carrying HBV preS2/S gene were constructed by homologous recombination in bacteria. The 293 cells, Vero cells, HepG2 cells and mesenchymal stem cells (MSCs) were infected with adenoviruses. The expressions of enhanced green fluorescent protein (EGFP) were observed with fluorescence microscope and the expressions of HBsAg were detected by RT-PCR and ELISA in vitro.</p><p><b>RESULTS</b>More than 90% of 293 cells, Vero cells, HepG2 cells or MSCs expressed EGFP after transfection at the MOI of 20 and the titers of HBsAg were more than 3.229 (A value) in culture supernatant.</p><p><b>CONCLUSION</b>The HBV preS2/S gene was not only expressed efficiently in immortalized cells, but also expressed efficiently in stem cells with the recombinant adenoviruses vector.</p>


Subject(s)
Adenoviridae , Genetics , Animals , Cell Line , Cell Line, Tumor , Chlorocebus aethiops , Enzyme-Linked Immunosorbent Assay , Gene Expression , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , Hepatitis B Surface Antigens , Genetics , Metabolism , Hepatitis B virus , Genetics , Allergy and Immunology , Humans , Microscopy, Fluorescence , Reverse Transcriptase Polymerase Chain Reaction , Transfection , Vero Cells
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