Cell Line, Tumor ; Enzyme Inhibitors ; pharmacology ; Heterocyclic Compounds, 1-Ring ; pharmacology ; Humans ; Proteasome Endopeptidase Complex ; drug effects

OBJECTIVETo explore the significance of HBV large protein (LHBs) in diagnosis of hepatitis B, we detected the LHBs, HBV DNA, PreS1 and other hepatitis B viral markers (HBV M) in the serum of patients infected with HBV.

METHODSHBV DNA was quantitatively detected using RT-PCR, LHBs, PreS1 and HBV M were analyzed by ELISA in totally 385 serum samples.

RESULTSThere was a significant difference between the positive rate of LHBs (86.97%) and PreS1 (49.5%) in the 307 serum positive for HBV DNA (P less than 0.05). There was a correlation between the levels of LHBs and the logarithm of HBV DNA (r=0.935). In the serum specimens of patients negative for HBeAg, there was no significant difference between the positive rate of LHBs (76.92%) and the HBV DNA (67.95%), but the positive rate of PreS1 (45.73%) was lower than that of LHBs or HBV DNA.

CONCLUSIONThere was a close correlation between the copies of HBV DNA and the levels of LHBs, both the positive rate and the coincidence rate of LBHs and HBV DNA were higher than those of PreS1. LHBs can reflect the replication status of HBV.

Adolescent ; Adult ; Aged ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B ; blood ; diagnosis ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Humans ; Male ; Middle Aged ; Protein Precursors ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Viral Matrix Proteins ; blood ; Young Adult

OBJECTIVEEpstein-Barr virus (EBV) is a common causative agent of infectious mononucleosis (IM) and capable of efficiently immortalizing primary B cells into continuously growing lymphoblastoid cells in vitro. As B cell activation antigen, CD23 expression is induced by EBV infection of B cells and remains constitutively expressed at high levels in virtually all EBV-immortalized cells, which have been strongly linked to the development of B-cell lymphoproliferative disease and lymphoma. Whereas previous studies were performed in vivo in animals or ex vivo cultures, the present study aimed to explore the role of EBV-immortalized cells (CD23(+)/CD19(+)) in vivo analysis of children with EBV-IM.

METHODSIn a prospective trial, a group of 30 patients with IM (18 boys and 12 girls) with mean age of 3.9 +/- 1.3 years (range 6 months to 8 years) were enrolled. Clinical diagnosis of IM was confirmed based on fever, lymphadenopathy, splenomegaly, lymphocytosis (> 50%), atypical lymphocytes (> 10%) in blood smears and the elevated levels of IgM antibody against EBV capsid antigen. The day of onset of fever was recognized as day 1 of illness. Blood samples taken during acute (3 - 5 days), early convalescent (about 11 - 15 days) and convalescent phase (about 30 - 45 days) were analyzed for expressions of CD19(+)/CD23(+), CD23, CD19 on peripheral blood mononuclear cells by flow cytometry (FCM) and was compared with those of control group.

RESULTS(1) The levels of CD23(+)/CD19(+) and CD23 expressions were markedly decreased in acute stage [CD23(+)/CD19(+) (2.22 +/- 1.47)%, (132 +/- 91)/mm(3); CD23 (3.12 +/- 1.88)%, (195 +/- 102)/mm(3)] and in early convalescent stage [CD23(+)/CD19(+) (4.51 +/- 2.25)%, (166 +/- 85)/mm(3); CD23 (5.55 +/- 2.76)%, (231 +/- 130)/mm(3)] in patients with IM as compared with those of the healthy controls [CD23(+)/CD19(+) (6.71 +/- 2.25)%, (215 +/- 68)/mm(3); CD23 (7.85 +/- 3.09)%, (249 +/- 86)/mm(3), respectively]. The earlier the history was, the lower the expressive levels were. The levels of CD23(+)/CD19(+) expressions returned to, but those of CD23 expressions exceeded, normal level in convalescent stage [CD23(+)/CD19(+) (6.72 +/- 2.16)%, (213 +/- 108)/mm(3); CD23 (9.46 +/- 2.73)%, (366 +/- 200)/mm(3)]. (2) There was a positive correlation in the expressions of CD23(+)/CD19(+) and CD23 among the three stages (P < 0.01). The positive correlation between the expressions of CD23(+)/CD19(+) and CD19 only occurred during acute stage (P < 0.01). There was no correlation between the expressions of CD23 and CD19 (P > 0.05).

CONCLUSIONEBV-immortalized cells and CD23(+) cells were inhibited effectively during the acute and early convalescent stage of IM. With the recovery of the disease, they gradually recovered and the levels of CD23 expressions exceeded normal level in convalescent stage.

B-Lymphocytes ; metabolism ; Cell Culture Techniques ; Cell Survival ; Child ; Child, Preschool ; Female ; Herpesvirus 4, Human ; pathogenicity ; Humans ; Infant ; Infectious Mononucleosis ; metabolism ; Male ; Receptors, IgE ; metabolism

OBJECTIVEInfectious mononucleosis (IM) is a lymphoproliferative disease caused primarily by the Epstein-Barr virus (EBV) infection. The initial viral infection by EBV occurs in B lymphocytes and is followed by an extensive proliferation of T lymphocytes. Previous studies on immunity to EBV (including IM) have mainly focused on activation of peripheral blood T cells, which are responsible for the lymphocytosis in blood during acute IM. B cells, regarding CD23 as their activation marker, are the target cells of EBV infection. There are few reports on their effect in patients with IM. The role of them during acute IM is not known yet. The present study aimed to explore the action of B cells in patients with IM.

METHODSIn a prospective trial, a group of subjects comprised 22 patients with IM (14 boys and 8 girls) with mean age of 3.48 +/- 0.81 years (range 7 months to 8 years). Clinical diagnosis of IM was confirmed based on fever, lymphadenopathy, splenomegaly, lymphocytosis (> 50%), atypical lymphocytes (> 10%) in blood smears and the elevated levels of IgM antibody against EBV capsid antigen. The day of onset of fever was recognized as day 1 of illness. Blood samples taken during acute (3 - 5 days) and convalescent phase (about 15 days) were analyzed for expressions of CD19, CD19(+)/CD23(+) on PBMC by flow cytometry (FCM) and was compared with those of control group. The number of the days with fever was recorded.

RESULTS(1) The levels of CD19 and CD19(+)/CD23(+) expressions were markedly decreased in acute stage [CD19 (5.63 +/- 2.91)%, (387 +/- 178)/mm(3), CD19(+)/CD23(+) (2.45 +/- 1.87)%, (160 +/- 99)/mm(3)] and in convalescent stage [CD19 (12.49 +/- 5.70)%, (428 +/- 156)/mm(3), CD19(+)/CD23(+) (5.05 +/- 2.79)%, (172 +/- 78)/mm(3)] in patients with IM as compared with those of the healthy controls [CD19 (16.20 +/- 2.80)%, (545 +/- 150)/mm(3); CD19(+)/CD23(+) (7.08 +/- 2.78)%, (249 +/- 136)/mm(3)]. The earlier the specimens were taken after onset, the lower the expressed levels were. (2) There was a positive correlation of the expressions of CD19 and CD19(+)/CD23(+) between acute and convalescent stage (P < 0.01);there was also a positive correlation between the expressions of CD19 and CD19(+)/CD23(+) during acute and convalescent stage (P < 0.01). (3) A negative correlation was found between the duration of fever and the level of CD19 and CD19(+)/CD23(+) in acute stage (P < 0.01).

CONCLUSIONThe results indicate that B cells and CD23(+) B cells were significantly inhibited during the onset of IM in the patients, that with the recovery of the disease, the condition was gradually improved, and that the more evidently the CD19 and CD19(+)/CD23(+) decreased, the more serious the clinical symptoms were and the longer time the recovery needed. The levels of CD19 and CD19(+)/CD23(+) expressions may be useful in diagnosis and predicting the severity.

Antigens, CD19 ; immunology ; B-Lymphocytes ; immunology ; Child ; Child, Preschool ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Infectious Mononucleosis ; diagnosis ; immunology ; virology ; Male ; Prospective Studies ; Receptors, IgE ; immunology ; T-Lymphocytes ; immunology

OBJECTIVETo explore the inhibition pathway of the EBV-immortalized cells (CD23(+)) in children with infectious mononucleosis (IM) caused by Epstein-Barr virus.

METHODSThe expressions of CD23, CD19, CD95, Bcl-2 and the co-expressions of CD23CD95, CD19CD23 on peripheral blood mononuclear cell (PBMC) were analyzed by flow cytometry (FCM) during acute phase, early convalescent phase and convalescent phase of 34 EBV-IM children and compared with that of 24 healthy donors.

RESULTS(1) The levels of CD23(+) and CD23(+)CD19(+) cells decreased and CD95(+), CD95(+)CD23(+), Bcl-2(+) cells increased markedly in IM patients in acute phase [CD95(+) cells (19.43 +/- 8.46)%; CD95(+)CD23(+) cells (1.81 +/- 1.71)%; Bcl-2(+) cells (23.41 +/- 26.47)%] and early convalescent phase [CD95(+) cells (12.94 +/- 5.05)%; CD95(+)CD23(+) (1.05 +/- 1.20)%; Bcl-2(+) cells (10.54 +/- 9.68)%], as compared with those of healthy controls [CD95(+) cells (10.39 +/- 2.90)%; CD95(+)CD23(+) cells (0.50 +/- 0.46)%; Bcl-2(+) cells (7.25 +/- 2.88)%]. The earlier the course of IM, the more abnormal the expressive levels. All the abnormal results returned to normal in convalescent phase. (2) Positive relationships were observed between the expressions of CD95(+)CD23(+) cells and that of CD23(+) cells, CD23(+)CD19(+) cells during acute and early convalescent phase, the expressions of Bcl-2(+), CD3(+) cells and CD23(+), CD23(+)CD19(+) cells during acute phase, the expressions of CD95(+)CD23(+) cells and Bcl-2(+) cells during acute phase, and the expressions of CD95(+)CD23(+) cells and CD95(+) cells during convalescent phase.

CONCLUSIONThe results indicate that CD95L-CD95 mediated apoptosis plays an important role in eliminating EBV-immortalized cells, which is counteracted partly by Bcl-2.

Antigens, CD19 ; blood ; Cell Transformation, Viral ; Cells, Cultured ; Child ; Child, Preschool ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Infectious Mononucleosis ; blood ; pathology ; virology ; Male ; Receptors, IgE ; blood ; bcl-Associated Death Protein ; blood ; fas Receptor ; blood

Objective To detect EIA factor expression in human rectal cancer and normal tissue and to determine whether it is correlated with invasion and metastasis of human rectal cancer. Methods Real- time RT-PCR was used to detect E1AF expression in matched rectal cancers and normal tissues from g6 in- patients.Results Among the 86 rectal cancer samples tested,55 cases E1AF mRNA overexpression was ob- served. The mRNA expression of E1AF in the sample group was remarkably different from that in the control group.In carcinomas,E1AF mRNA expression correlated significantly with histological type,depth of inva- sion,lymph node and distant metastasis and advanced Duke stage.Conclusion E1AF is correlated signifi- cantly with invasion and metastasis of human rectal cancer and may be an important factor in the invasion and metastasis.

0.05) and the two methods had good correlation(r=0.822).CONCLUSIONS The method of FCST established by as in this study is simple,repeatable,with high accuracy and easy to determine MIC and has good application prospects in clinical antifungal susceptibility testing.

Objective:To establish a polyethylene glycol ( PEG6000 ) precipitation method for screening macroprolactinemia in patients with high serum prolactin ( PRL) . Methods:PEG6000 precipitation method was used to remove macroprolactin ( MPRL) molecules in serum of PRL-elevated patients.The effect of PEG6000 precipitating serum MPRL was determined by Sephacryl S-100 HR chromatography plus chemiluminescent immunoassay and SDS-PAGE plus Western Blot assay .The PEG6000 precipitation plus chemiluminescent immunoassay was applied to screen serum samples of PRL-elevated patients for macroprolactinemia .The clinical manifestations of patients with true-hyperprolactinemia , hyperprolactinemia/macroprolactinemia or true-macroprolactinemia were analyzed and compared .Results: After precipitation with PEG6000 , MPRL peak or hybridization signal in the serum samples was markedly decreased, while the big or small prolactin (BPRL or SPRL) levels were not affected. In 1538 PRL-elevated patients , 16 .1% ( 247/1538 ) were detectable for macroprolactinemia , while the 83 .9% ( 1291/1538 ) were identified as true-hyperprolactinemia .In 247 samples of macroprolactinemia , 93 .5% ( 231/247 ) were determined as true-macroprolactinemia, while 6.5% ( 16/247 ) were identified as hyperprolactinemia plus macroprolactinemia .In 508 true-hyperprolactinemia patients , menoxenia , menolipsis/menostasia , dysgenesia or hypophysoma were manifested in 438 (86.2%), which were also manifested in 85.7% ( 6/7 ) of hyperprolactinemia/macroprolactinemia patients .However , only 11 cases in 71 true-macroprolactinemia patients ( 15 .5%) presented above clinical diseases .Conclusion: There is a certain proportion of true-macroprolactinemia ( pseudo-hyperprolactinemia ) in serum PRL-elevated patients.The PEG6000 precipitation method established in this study can efficiently distinguish true-hyperprolactinemia from pseudo-hyperprolactinemia in patients .

Objective To evaluate the value of plasma ProGRP,CYFRA 21-1 and CEA in patiens with lung cancer.Methods The levels of plasma ProGRP,CYFRA 21-1 and CEA were detected in 85 healthy control,49 benign lung diseases and 143 lung neoplasms.The levels of ProGRP in the patients with SCLC was monitored.Results The level of plasma ProGRP in SCLC ( M 179.1 ng/ml) was significantly higher than adenocarcinoma ( M 35.3 ng/ml),squamous-cell carcinoma ( M 33.3 ng/ml),healthy control (M 35.6 ng/m)and benign lung diseases (M 33.3ng/m ),P ＜ 0.001.The sensitivity and specificity for diagnosing SCLC by ProGRP were 60.6％ and 95.0％ respectively.In the effective treatment group,ProGRP reduced 45.9％,in the progression group,ProGRP increased 103.1％,P ＜ 0.05.The level of CEA in the metastatic adenocarcinoma (M 10.22 ng/ml ) was significantly higher than non-metastatic adenocarcinoma ( M 3.85ng/ml) and squamous cell carcinoma ( M 2.56 ng/ml) ( P ＜ 0.01 ).Conclusion The plasma ProGRP is a good indicator for diagnosing and evaluating cure effect in SCLC; the high expression of CEA is related to the metastatic adenocarcinoma.

This study was aimed to investigate the changes of reactive oxygen species and antioxidative capacity on nitric oxide induced apoptosis in HL-60 cells. By means of in vitro incubation of HL-60 cells with sodium nitroprusside (SNP), the growth inhibition was detected by MTT assay. Cell morphology was observed by transmission electronmicroscopy and light microscopy. The apoptosis was analyzed by DNA agarose gel electrophoresis, DNA content and Annexin-V/PI labeling method. Reactive oxygen species (ROS) labeled with dihydrorhodamin 123 in cells was determinated by flow cytometry. The SNP-treated cells were examined for glutathione (GSH) level and activity of catalase (CAT), glutathione S-transferase (GST) and glutathione peroxidase (GPX). The results indicated that SNP could inhibit HL-60 cell growth. Cell apoposis was confirmed by typical cell morphology, DNA fragment, sub-G(1) phase and Annexin-V/PI labeling method. HL-60 cell apoptosis was induced by SNP in a dosage- and time-dependent manner. After exposing to SNP at the concentration of 0.5 - 3.0 mmol/L for 48 hours, the mean fluorescence intensity of ROS in cells was significantly higher than those in groups control and potassium ferricyanide (PFC). During the apoptosis process, level of ROS in cells increased, levels of GSH, CAT, GPTand GPX decreased. The significant dose-effect relationship existed between the levels of ROS, CAT, GST, GPX and SNP dose. It is concluded that change of intracellular reactive oxygen species and antioxidative capacity are an important factors during the process of SNP-induced apoptosis in HL-60 cell.
Antioxidants ; metabolism ; Apoptosis ; drug effects ; Cell Nucleus ; drug effects ; ultrastructure ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; HL-60 Cells ; Humans ; Microscopy, Electron ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Reactive Oxygen Species ; metabolism