Antiviral Agents ; therapeutic use ; Cytokines ; metabolism ; Dendritic Cells ; immunology ; metabolism ; pathology ; Endotoxemia ; complications ; pathology ; Hepatitis B ; complications ; pathology ; Humans ; Hypoxia ; complications ; pathology ; Ischemia ; complications ; pathology ; Liver ; metabolism ; pathology ; Liver Failure ; etiology ; immunology ; pathology ; therapy ; T-Lymphocytes ; immunology ; pathology

Antiviral Agents ; therapeutic use ; Hepatitis B, Chronic ; drug therapy ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferons ; therapeutic use ; Treatment Outcome

Adult ; Hepatitis C ; etiology ; therapy ; Humans ; Kidney Transplantation ; Male ; Recurrence

OBJECTIVETo construct DNA double-strand break (DSB) repair protein hKu70 deficient cell strain and to observe its biological characters for studying the functions of hKu70 gene and the effects of occupational harmfulness factors on DSB repair.

METHODSHuman lung fibroblasts (HLF) were transfected with the eukaryotic expression plasmids of hKu70 gene antisense RNA (pEGFP-C1-K) to construct hKu70 protein deficient cells (named as "HLFK"). The protein expression levels of hKu70 gene in HLFC and HLFK were detected by the Western blotting to estimate the effects of antisense inhibition. Morphology, growth character and growth status in soft agar of transfected HLFK were observed.

RESULTSpEGFP-C1-K vector was successfully expressed in HLF. The protein expression level of hKu70 gene in HLFK was decreased by 42% as compared with that in HLFC. No obvious changes of the biologic characters were observed in HLFK.

CONCLUSIONThe hKu70 protein deficient cell strain was successfully constructed. The hKu70 protein deficiency alone didn't induce obvious changes of the biological characters in HLFK.

Antigens, Nuclear ; analysis ; Cell Division ; DNA Damage ; DNA Helicases ; DNA Repair ; DNA-Binding Proteins ; analysis ; deficiency ; Humans ; Ku Autoantigen ; RNA, Antisense ; Transfection

OBJECTIVETo construct pEGFP-C1-T vector, an eukaryotic expression plasmid of hMTH1 gene antisense RNA.

METHODSThe conservative region of hMTH1 gene was amplified by RT-PCR after total RNA being extracted from human embryo lung fibroblast (HLF) and then cloned into pGEM-T vector. After the recombinant plasmid was certified by DNA sequencing, the conservative region of hMTH1 gene was inserted into pEGFP-C1 vector reversedly and pEGFP-C1-T vector was constructed. The efficiency of antisense inhibition was verified by Western blotting after cell transfection.

RESULTS423 bp fragment including conservative region of hMTH1 gene was obtained by RT-PCR. After cloned by pGEM-T vector and certified by DNA sequencing, pEGFP-C1-T vector was successfully constructed by means of recombinant DNA technology. Additionally pEGFP-C1-T vector could efficiently decrease hMTH1 protein level by 46%.

CONCLUSIONThe efficient expression vector of hMTH1 gene antisense RNA, pEGFP-C1-T has been constructed successfully.

DNA Repair Enzymes ; Genetic Vectors ; genetics ; Humans ; Phosphoric Monoester Hydrolases ; genetics ; Plasmids ; RNA, Antisense ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the status of vaccination against hepatitis B among postgraduate students of medical institutions of higher education in Guangzhou.

METHODSHBsAg and anti-HBs in the serum samples from 1139 postgraduate students were detected by ELISA. Data on hepatitis B vaccine inoculation were investigated by using a questionnaire. Statistical analyses were performed by using SAS software.

RESULTSThe HBsAg positive rate among the 1139 postgraduate students was 2.90 percent. The HBsAg positive rates in hepatitis B vaccine inoculated (1.15 percent) and non- inoculated (21.69 percent) postgraduate students were significantly different (x2=119.11, P<0.0001). The positive rates of HBsAb between the two groups were also significantly different (x2=62.05, P<0.0001). Among the hepatitis B vaccine inoculated students, 17.31 percent were negative for HBsAb. The positive rate of HBsAb among those inoculated the vaccine within the past 3 years was higher than that among those inoculated the vaccine earlier (0-3 years vs. 4-6 year, P=0.0089) (0-3 years vs. 7-9 years, P=0.0172) (0-3 years vs. >9 years, P=0.0474). The positive rate of HBsAb among the students who received hepatitis B vaccine booster dose was higher than that of the students who did not receive any booster dose (P=0.0093).

CONCLUSIONWith the increase of ages, the effect of vaccination for hepatitis B decreased. Male populations may be more susceptible to hepatitis B virus than female. It is necessary to monitor HBsAb levels for those who were inoculated with HBV vaccine more than 3 years ago to give booster dose in time to prevent HBV infection.

Adult ; China ; Female ; Hepatitis B ; immunology ; prevention & control ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Hepatitis B virus ; immunology ; Humans ; Male ; Middle Aged ; Students, Medical ; Surveys and Questionnaires ; Vaccination ; Young Adult

Hepatitis C ; etiology ; virology ; Hepatitis Viruses ; Humans ; Liver Transplantation ; Recurrence

OBJECTIVETo optimize cultivation methods of bone marrow mesenchymal stem cells (MSCs) from hepatitis B patients and to investigate their biological characteristics.

METHODSGrowth curves of hepatitis B patients MSCs cultivated with five culture media and two inoculation methods were compared; the shapes, appearances, surface markers and bionomics of the cultivated MSCs were studied.

RESULTSInoculating the cells obtained directly from bone marrow aspirations was not as successful as using the marrow cells after their density gradient centrifugations (76% vs 88%), but the differences in the results were not statistically significant (P more than 0.05). The successful cultivation rates using five culture media were different and the differences were statistically significant (P less than 0.01). The autoserum medium was most successful, fatal bovine serum (FBS) medium was next successful and the non-patient serum medium was the least successful. The growth curves of the cultivations using the different media were parallel to this. Changing the whole culture media every 2 or 3 days was better than changing half of the media. The shapes, appearances, surface markers and the growth characteristics of MSCs from the hepatitis B patients were almost the same as MSCs from the normal adult.

CONCLUSIONThe best cultivation method of MSCs from hepatitis B patients is: separating marrow cells using density gradient centrifugal separation, cultivating them using an autoserum culture medium, and completely changing the medium every 2-3 days. The biological characteristics of MSCs from the hepatitis B patients using the above methods are almost the same as those from normal adults.

Adult ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cells, Cultured ; Culture Media ; Hepatitis B ; Humans ; Mesenchymal Stromal Cells ; cytology ; Middle Aged

OBJECTIVETo investigate the immune effects of three different programs for revaccination among adults of non- and hypo-responders to recombinant Hepatitis B vaccine.

METHODSThose who were once immunized with recombinant Hepatitis B vaccine more than one standard schedule (0, 1, and 6 months) in two years and negative for Hepatitis B markers were randomly given three-different projects for revaccination. 34 adults of A group were given GM-CSF 300 microg by subcutaneous injection for the first day, then 10 microg each time by intramuscular route for routine immune method. 33 adults of B group were given Hepatitis B vaccine 20 microg each time. 33 adults of C group were given Hepatitis B vaccine 10 microg each time. The blood samples were collected before the first injection and in 1, 2 and 8 months following the first injection to test Anti-HBs.

RESULTSAt T1, the anti-HBs positive conversion rate of group A, B and C was 26.47%, 48.48% and 18.18% respectively (chi-2 = 7.20, P = 0.027). At T8, the anti-HBs positive conversion rate of group A (64.71%) and group B (75.76%) were higher than group C (39.39%), and there was significant difference (chi-2 = 9.07, P = 0.011). At T1, the anti-HBs level of group B (417.00 +/- 69.36) was higher than that of group A (203.74 +/- 79.56). At T2, the anti-HBs level of group B (458.17 +/- 64.09) was higher than that of group C (257.86 +/- 76.60). At T8, the anti-HBs level of group A (501.48 +/- 70.00) and group B (532.73 +/- 68.82) were higher than those of group C (256.12 +/- 75.39) (t =4.27, P = 0.0173).

CONCLUSIONSchemes of augmentation doses of Hepatitis B vaccine and being combined with GM-CSF should be in effect for non- and hypo-responders to Hepatitis B vaccine.

Adolescent ; Adult ; Antibody Formation ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Humans ; Male ; Middle Aged ; Young Adult

Humans ; Intestines ; immunology ; microbiology ; Liver Diseases ; immunology ; microbiology