To explore the effect of a tyrosine-kinase inhibitor STI571 and P21(WAF) gene clone on the proliferation, cycle, apoptosis of leukemia cell line K562, P21(WAF) gene was obtained by RT-PCR, and its sequence was approved to be correct, then P21-pcDNA3.1 vector was constructed and transfected into K562 cell line. After selected with G418, P21-pcDNA3.1-K562 cell clone that stably expression P21(WAF) was isolated. P21(WAF) protein was identified by Western blot. The survival rate were tested by MTT. Cell cycle and apoptosis were tested by flow cytometry. The results showed that the expression of P21(WAF) protein could be detected by Western blot in P21-pcDNa3.1-K562 cells. A strong inhibition of cell proliferation was observed in P21-pcDNA3.1-K562 cells as compared with that of the control. The cells cycle were arrested in G(0)/G(1) phase. The percentage of apoptosis was declined slightly after P21-pcDNA3.1-K562 cells were combined with STI571, meanwhile its survival rate declined more slowly than that of K562 cell with STI571. In conclusion, P21(WAF) inhibits the proliferation of K562 cell, meanwhile slightly inhibits its apoptosis induced by STI571and decrease its sensitivity to STI571.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; genetics ; Base Sequence ; Benzamides ; Blotting, Western ; Cell Cycle ; drug effects ; genetics ; Cell Proliferation ; drug effects ; Cloning, Molecular ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Humans ; Imatinib Mesylate ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; pathology ; Molecular Sequence Data ; Piperazines ; pharmacology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; pharmacology ; Transfection

Objective To observe the status of occuh hepatitis B virus infection in chronic viral hepatitis patients with non-A to E hepatitis virus infection and explore the diagnostic value of fluorescence quantitative polymerase chain reaction (FQ-PCR) technique for occult hepatitis B virus infection. Methods The amount of HBV-DNA in serum and liver tissue from 57 patients with non-A to E hepatitis virus infection who were diagnosed as chronic viral hepatitis by Menghini method liver biopsy were detected by using FQ-PCR technique,then the relation between the viral load of HBV DNA in liver tissue and hepatic inflammatory activity were analyzed.Results Thirteen (22.81% ),22 (38.60%) patients were positive for HBV DNA in serum and liver tissue,respectively.The positive rate and the level of HBV DNA quantity in liver tissue were significantly higher than those in serum;HBV DNA was found positive in both serum and liver tissue in 13 cases,negative in both serum and liver tissue in 35,positive in liver tissue but negative in serum in 9,and in none of the cases HBV DNA was positive in serum but negative in liver tissue ( P<0.01 ).The logarithmic value of HBV DNA from 13 patients in liver tissue and in serum was respectively:(6.62±1.21 ) copies/g vs.(4.03±1.06) copies/ml,P < 0.01.The hepatic lesions of all HBV DNA positive patients were active pathologic changes,but the level of HBV DNA in liver tissue was not significantly correlated with the grade of hepatic inflammation activity ( P0.05 ).Conclusion Occult HBV infection is the etiology of part of the chronic viral hepatitis patients with non-A-E hepatitis virus infection.Missed diagnosis will occur if diagnosis of hepatitis B is only hased on detection of serum HBV markers. It is useful for improvement of the diagnostic level of HBV infection via detection of HBV DNA quantitatively in serum especially in liver tissue of chronic viral hepatitis with non-A-E hepatitis virus infection by using FQ-PCR technique The chronic viral hepatitis patients occult HBV infection should be also given effective anti-viral therapy.

OBJECTIVETo observe the effect of hepatitis C virus (HCV) superinfection on the short-term and long-term hepatic pathological changes in patients with chronic hepatitis B (CHB).

METHODSHCV-RNA of twice corresponding period serum samples was detected via reverse transcription polymerase chain reaction assay from 230 patients with CHB for whom liver biopsy was performed at an interval of 0.5-15 years, respectively. The hepatic pathological changes of the patients with CHB who were serum HCV-RNA positive at the beginning of observation and persistently positive between the starting and ending of observation were respectively compared with those of serum HCV-RNA negative and persistently negative patients.

RESULTS41 patients (17.83%) were positive for serum HCV-RNA at the beginning of observation. There were significant differences in the severity of hepatic inflammatory activity grade and fibrosis stage between serum HCV-RNA positive and negative patients with CHB (P < 0.05). Twenty-nine patients were persistently positive for serum HCV-RNA in the beginning and end of observation. Compared with persistently negative patients who were 116 patients selected from the above-mentioned 230 patients and they were comparable with HCV-RNA persistently positive patients in mean follow-up time, age and sex, the long-term progression of hepatic inflammatory activity grade and fibrosis stage in persistently positive patients were more speedy (P < 0.01).

CONCLUSIONHCV superinfection worsens the hepatic pathological changes of patients with CHB and speeds up its progression.

Adult ; Female ; Hepacivirus ; physiology ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; blood ; pathology ; virology ; Host-Pathogen Interactions ; Humans ; Liver ; pathology ; virology ; Male ; RNA, Viral ; genetics ; Retrospective Studies ; Superinfection ; virology ; Time Factors ; Viral Load

This study was aimed to investigate the role of mitochondria pathway in signal transduction of chronic myeloid leukemia (CML). After bcr3/abl2 antisense oligodeoxynucleotide (ASO) was introduced into CML cell line K562 cells by liposomal transfection, the cell viability was detected by MTT assay, the cell apoptosis was determined by flow cytometry (FCM), the mitochondrial membrane potential (DeltaPsi) was labeled by Rhodamine 123 and examined by FCM, and the expression of mitochondrial apoptosis signal transduction pathway related proteins cytochrome C was analyzed by Western blot. The results showed that after K562 cells were exposed to 2 micromol/L of bcr3/abl2 ASO for 24 hours, bcr3/abl2 ASO significantly inhibited cell viability with inhibitory rate of 65.7%, induced the apoptosis of K562 cell line with apoptotic rate of 16.9%, and decreased mitochondrial Deltapsi of K562 cells with the reducing rate of 38.33%, enhanced the expression of cytochrome C with increase of optical density value from 2.33 +/- 0.3 to 4.78 +/- 0.1 by laser photometric scanning. It is concluded that mitochondria pathway plays an important role in signal transduction of chronic myeloid leukemia by directing apoptotic signal transduction.
Apoptosis ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Membrane Potential, Mitochondrial ; Membrane Potentials ; Mitochondria ; metabolism ; Oligonucleotides, Antisense ; Signal Transduction