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Biomedical subjects

M Zhan

Publications and source records attributed to M Zhan.

28 records · Page 2Linked to original sources
PubMed1999

Easy detection of all T cell receptor gamma (TCRG) gene rearrangements by Southern blot analysis: recommendations for optimal results.

Southern blot analysis of T cell receptor (TCR) gene rearrangements has proven to be a helpful tool to establish clonality in T cell leukemias and lymphomas. To improve the detection of clonal TCR gamma (TCRG) gene rearrangements by Southern blot analysis, we designed four new Jgamma probes and determined the most optimal restriction enzymes to be used with these probes. Based on detailed analysis of the sequences as well as on hybridization experiments with the TCRGJ21 probe, the Jgamma1.2 and Jgamma2.1 downstream areas were found to be highly homologous, suggesting that during evolution the duplication of the Jgamma region was followed by deletion of the tentative Jgamma2.2 gene segment. Southern blot analysis of 51 T cell acute lymphoblastic leukemias (T-ALL) revealed that all TCRG gene rearrangements can be detected by use of the TCRGJ13 probe in EcoRI digests and the TCRGJ21 probe in PstI digests. Additional probes and digests allow a more precise identification of the exact type of TCRG gene rearrangements in the majority of cases. Almost 90% of the TCRG gene rearrangements in T-ALL involved the Jgamma2 region (16% Jgamma2.1 and 72% Jgamma2.3), whereas Jgamma1 region rearrangements were particularly found in TCRgammadelta+ T-ALL. This information has implications for design of primer sets for PCR analysis at diagnosis and for PCR target choice in detection of minimal residual disease during follow-up of T-ALL patients.

Blotting, Southern↗
PubMed1999

Schedule-dependent reversion of cisplatin resistance by 5-fluorouracil in a cisplatin-resistant human lung adenocarcinoma cell line A549DDP.

OBJECTIVE: To study the schedule-dependent reversion of cis-diamminedichloroplatinum (CDDP) resistance by 5-fluorouracil (5-Fu) in a CDDP resistant human lung adenocarcinoma cell line A549DDP. METHODS: Dimethylthiazol dipheryltetrazolium bromide (MTT) assay and immunocytochemistry were used. RESULTS: After the A549DDP was treated with CDDP, followed immediately by exposure to 5-Fu, cytotoxicity of CDDP increased 1.8 fold. After pretreatment of A549DDP with 5-Fu, followed immediately by exposure to CDDP, the cytotoxicity of CDDP increased 3.9 fold. After pretreatment of A549DDP with 5-Fu, after a 24- or 48-hour drug-free interval, followed by exposure to CDDP, the cytotoxicity of CDDP increased 20 and 250 fold, respectively, and the A549DDP was rendered more sensitive than its parental cell line A549. In parallel with the increased cytotoxicity, the cellular GSH content was significantly reduced at 24 or 48-hour after 5-Fu pretreatment. However, depletion of GSH by buthionine sulfoximine (BSO) only resulted in partial reversion of CDDP resistance. 5-Fu could also inhibit the expression of MRP, but had no effect on the expression of GST pi. The effect of 5-Fu on the parental cell line A549 was much smaller than that in A549DDP. CONCLUSION: Scheduled administration of 5-Fu can reverse CDDP resistance completely through reduction of GSH and inhibition of MRP expression.

Adenocarcinoma↗
PubMed1999

cDNA cloning and sequence analysis of hepatitis G virus genome isolated from a Chinese blood donor.

OBJECTIVE: To obtain full-length sequence of a Chinese hepatitis G virus (HGV) strain (HGVch) and investigate the genetic characteristic of HGVch and its identity to other isolates. METHODS: Reverse transcription (RT) and nested-PCR were used to screen HGV RNA positive serum and amplify cDNA fragments. A positive serum without known hepatitis virus markers was selected for isolating HGV RNA template. The HGV genome was divided into 12 overlapping fragments and directly cloned into pGEM-T vector. Sequences were determined by dideoxy terminus-end method of DNA sequencing and then analyzed by computer. RESULTS: The twelve fragments of HGVch cover 9213 nucleotides in length, containing a large open reading frame (ORF) encoding 2873 animo acids polyprotein that began with a methonine residue and ended at termination codon. HGVch is about 86.5%-89.5% identical to other known HGV isolates at the nucleotide level and about 93.9%-96.2% at the deduced animo acid level. CONCLUSION: HGV is a non-A-E hepatitis causal agent, proved to be related with posttransfusion hepatitis in all over the world. Chinese HGV isolate has very close relationship to other isolates from Africa, Europe, Japan, without significant difference across the entire genome. It is suggested that the sequences of HGV isolates are very conservative and the evolution is very slow.

Blood Donors↗
PubMed1999

[Clinical significance of multidrug resistance-associated protein(MRP) gene expression in non-small cell lung cancer].

OBJECTIVE: To investigate expression of multidrug resistance-associated protein (MRP) in non-small cell lung cancer and its relation to histological type, TNM staging and prognosis. METHODS: In situ hybridization was used to examine mRNA expression of MRP. RESULTS: The overall positive rate of MRP expression was 74.1%, with 73.3% and 72.0% in adenocarcinoma and squamous-cell carcinoma, respectively. The expression of MRP was not related to histological subtypes, TNM staging and cell differentiation. In 47 patients who received chemotherapy, 35 patients with positive MRP expression(+(-)++) showed worse prognosis than in those with negative expression (P < 0.05). The median survival time was 8.7 months and 21 months in patients with positive and negative MRP expession, respectively. In patients with squamous-cell carcinoma, the survival rate was significantly lower in patients (n = 12) with positive MRP expression than in those (n = 7) with negative MRP expression (P < 0.05). Their median survival time was 6 months and 19.5 months, respectively (P < 0.05). CONCLUSION: The expression of MRP gene is negatively correlated with survival of patients with squamous-cell carcinoma, but not adenocarcinoma, who received chemotherapy.

Adult↗
PubMed1998

[Cloning and sequencing of cDNA in NS3 region of Chinese hepatitis G virus isolated from Henan Province].

To study the genetic characteristics of non-structural region 3 of HGV in China, a HGV-RNA positive sample was collected from Henan. HGV RNA was extracted and amplified by RT-nested-PCR. We got four overlap fragments and they were cloned into plasmid vector pcDNA II. The cDNA was sequenced by the Sanger's method. Nucleotide homology between Henan strain and other reported HGV isolates (PNF2161, R10291, GBV-C) was 85.7% to 93.8% over the 2137 nucleotides examined. Comparison of amino acid sequence deduced from Henan strain with that of other isolates showed 96.1% to 97.7% homology. Three amino acid variations were found in NS3 region of all the three Chinese isolates of HGV. In addition, a N-glycosylation site was found in the NS3 region of all the six isolates.

Amino Acid Sequence↗
PubMed1998

[The clinical and enzymatic changes in patients with viral hepatitis G infection].

For the purpose of making sure the clinical significance of hepatitis G virus, RT-nested PCR was applied to detect HGV RNA in 165 hepatitis patients, which included 24 acute hepatitis, 78 chronic hepatitis, 18 hepatitic cirrhosis, 4 hepatocellularcarcinom and 41 HBV and HCV carriers. The results showed that the infection of HGV existed in all kinds of hepatitis patients. Among the acute hepatitis 12.5% (3/24) was HGV RNA positive. 19 (24.4%) cases were HGV RNA positive in chronic hepatitis, among which 4 cases were simply HGV RNA positive (5.13%). The serum ALT level in 3 cases of simple acute HGV patients was between 488 +/- 65 U/L, the value of AST between 452 +/- 71 U/L, the TBiL at about 77.1 +/- 14.3 mumol/L. All these showed that only HGV infection could lead to acute hepatitis. The rising enzyme dropped to normal about a month later in acute hepatitis while HGV RNA would remain. The problem whether HGV infection is caused by simple acute and chronic hepatitis infection is under investigation.

Adolescent↗
PubMed1997

[Core protein of hepatitis C virus expressed in transgenic mice is associated with the expression of Fas molecule].

Hepatitis C virus (HCV) is a leading cause of morbidity and mortality worldwide but the pathogenesis of liver cell injury and the molecular mechanism of persistent infection of HCV are not well understood. The aim of this work is to establish the animal model of HCV infection for studying the biosynthesis and pathogenesis of HCV structural protein in transgenic mice. The structural genes (C+ E1 + E2) and 5'-uncoding region gene (5'UCR) of HCV isolated from China was inserted into pcDNA3 vector. The transgenic mice were produced by microinfecting this plasmid into mouse (C57BL/6 x ICR) embryos and the integrated mice were identified. The expression of core protein of HCV and mFas molecule were confirmed by immunohistochemical analysis using monoclonal antibody. The results revealed that the expression of core protein was detected in many mouses' tissues with nuclear staining, while the heart tissue was with cytoplasmic staining. The expression level in tissues was indicated as: heart>lung>kidney>liver. At meantime, Fas molecule was detected in heart, kidney and liver tissues, and the expression of Fas was parallel to that of core protein. Transgenic mouse containing HCV structural gene was established and the expression of Fas molecule may be associated with the expression of core protein of HCV. The Fas system or other signaling pathway to the programmed cell death may play important roles in HCV infection. Our transgenic mouse will provide a useful animal model to critically address issues of immune pathogenesis, direct cytopathic potential and carcinogenesis of the structural protein of HCV.

Animals↗
PubMed1997

[Dynamic changes of serum antibody titer and liver ultrastructural pathology in acute sporadic hepatitis E virus infected patients].

7 acute hepatitis E patients were tested for anti-HEV IgM and IgG antibodies by EIA. It revealed that anti-HEV IgM and IgG antibody titers were the highest at day 10-45 of the illness, and anti-HEV IgM declined gradually and totally lost within 2 months. Anti-HEV IgG had a similar condition to anti-HEV IgM, but at 7th month there still had 3 cases (40%) remained to be antibody positive. One case was examined for ultrastructural pathology. The swelling of liver cells on the fortieth day of illness was still to be found, the vacuolation of cytoplasm and condensation of mitochondria were present in liver cells.

Acute Disease↗
PubMed1997

[Diagnosis of hepatitis E virus infection in sporadic acute hepatitis].

The solid-phase enzyme immunoassay (EIA) was used for detecting anti-HEV immunoglobulin G (IgG) and M (IgM) by using a recombinant chimeric antigen (ORF3-2) from the open reading frame (ORF) 2 and 3 of hepatitis E virus gene. These sera were collected from the patients with sporadic acute viral hepatitis. The positive rate for anti-HEV IgG was 15.22% (7) among 46 patients with acute hepatitis, 5 of the 7 IgG positive sera were also positive (71.4%) for anti-HEV IgM. 91.8% (93) and 76.4% (78) were positive separately for anti-HEV IgG and IgM among 102 patients of suspected HEV infection (these patients were negative for anti-HAV, anti-HBV and anti-HCV), about 2/3 patients of HEV infection detected were simultaneously HEV-IgG and IgM positive. In control group of 98 normal persons, 4 (4.08%) were positive for anti-HEV IgG, but none were IgM positive. Paired sera of the patients in the acute phase and convalescent phase were detected, the titers of anti-HEV IgG with 4-16 fold decrease were seen in 6 cases and with 2-fold rise in 1 case.

Acute Disease↗