OBJECTIVETo explore the effects of hepatitis B virus X gene and p53 on hepatocellular growth.

METHODSTwo kinds of plasmids containing sense and antisense human wild p53 gene respectively were constructed. SMMU-7721 cells were transfected with HBx, sense-wtp53 antisense-wtp53 separately or cotransfected with either HBx and sense-wtp53 or HBx and antisense-wtp53. Flow cytometry was adopted to measure the apoptosis rates and the effects of HBx on cell cycle progression. The activity of p21(Waf1) promoter-luciferase construct was detected. Growth curves for SMMU-7721 stably transfected with pcDNA3 and pcDNA3HBx were analyzed.

RESULTSAfter doxorubicin administration, HBx was noticed able to initiate apoptosis of the liver cells. The apoptosis rate was 5.32% in the pcDNA3 transfected and 12.66% in the pcDNA3HBx transfected groups respectively. HBx could also abrogate p53-mediated apoptosis. The apoptosis rate in groups transfected with pcDNA3, pcDNA3wtp53 and pcDNA3HBx + pcDNA3wtp53 was 5.32%, 11.72% and 4.67% respectively. In compared with the normal group, the number of cells in transiently HBx-expressed group and HBx-transfected group decreased 4.79% and 10.25% respectively. HBx inhibited the activity of p21(Waf1) promoter-luciferase constructed (P < 0.05) and promoted cell growth. The growth rate of HBx expression cells was faster.

CONCLUSIONUnder DNA damage, HBx reduced expression of p21(Waf1) by repressing the activity of p53 protein, followed by disturbing the regulation of G(0)-G(1) cell cycle checkpoint, and promoted the growth rate of hepatoma cells.

Apoptosis ; Carcinoma, Hepatocellular ; pathology ; virology ; Cell Division ; Cell Line, Tumor ; Genes, p53 ; Hepatitis B Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; pathology ; virology ; Trans-Activators ; biosynthesis ; genetics ; Transfection ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVETo investigate the expression of five kinds of cyclins in hepatocellular carcinoma (HCC) and their association with degree of tumor differentiation, metastasis and infection of hepatitis B virus (HBV).

METHODSThe HCC tissue microarrays were composed of those from 273 cases of HCC tissues, 144 surrounding-tumor liver tissues and 10 normal liver tissues obtained from autopsy. The diameter of each specimens on tissue microarrays was 2.0 mm. Immunohistochemistry was used to detect the expression of cyclin A, cyclin B, cyclin D1, cyclin D3 and cyclin E on HCC tissue microarrays. The association of the expression of these cyclins with the infection rate of HBV was also analyzed.

RESULTSThree paraffin-embedded HCC tissue microarrays were successfully constructed, including 136, 143 and 148 tissue spots, respectively. The positive rates of cyclins in 273 cases of HCC were cyclin A 52.7%, cyclin B 45.4%, cyclin D1 35.9%, cyclin D3 44.3% and cyclin E 23.1%; while the figures in 144 surrounding-tumor tissues were 8.3%, 5.6%, 4.9%, 6.3% and 1.4%, respectively. In 10 normal liver tissues these cyclins exhibited negative staining, with the exception that cyclin D1 was positive in one case of normal liver tissue. The positive rate of cyclins in HCC were significant higher than those in surrounding-tumor liver tissues (P < 0.01), in HCC tissues with histological grade II and III, the cyclins expression were stronger than that in grade I (P < 0.05). The positive rates of cyclins, except cyclin A in HCC with portal vein invasion were higher than those without portal vein invasion (P < 0.01). Infection of HBV did not have significant relationship with the expression of cyclins (P > 0.05).

CONCLUSIONCyclins in different cell cycles overexpressed at varied levels in hepatocellular carcinoma, and the increased expression of cyclins may shorten the tumor cell cycle phase, accelerate cell proliferation, and have a close relationship with HCC aggressiveness.

Carcinoma, Hepatocellular ; chemistry ; Cyclin A ; analysis ; Cyclin B ; analysis ; Cyclin D1 ; analysis ; Cyclin D3 ; Cyclin E ; analysis ; Cyclins ; analysis ; Hepatitis B ; metabolism ; Humans ; Immunohistochemistry ; Liver Neoplasms ; chemistry

OBJECTIVE: To determine the characteristics of a novel gene Mip5 (GenBank accession number AY553870) and its expression under physiological and pathological conditions. METHODS: The characteristics of Mip5 were analyzed by bioinformatic programs including BLAST, spidey, psort, ClustalW and so on. RT-PCR was performed to detect Mip5 expression. RESULTS Bioinformatic analysis showed that Mip5 gene lied in the 13th chromosome and contained 8 exons and 7 introns, its open reading frame contained 909 bp and its protein production was 302 amino acid residues including 6 kelth domains. Under normal conditions, MIP5 expressed abundantly in the heart, brain and kidney, but its expression could not be detected in the liver and muscle. Expression of Mip5 gene was increased significantly after ischemia-reperfusion compared with the sham groups, and reached its peak at 3 h and recovered at 12 h after the reperfusion. Conclusion Mip5 gene is a novel gene containing a putative open reading frame of 302 amino acids residues and may play an important role in rat cardiomyocytes suffering ischemia processing.
Amino Acid Sequence ; Animals ; Base Sequence ; Chromosomes, Human, Pair 13 ; genetics ; DNA, Complementary ; genetics ; Humans ; Male ; Molecular Sequence Data ; Myocardial Ischemia ; genetics ; Myocardial Reperfusion Injury ; genetics ; Open Reading Frames ; genetics ; Rats

Objective To investigate the quality of life (QOL) and mental health status of patients with HIV/AIDS in Minhang District of Shanghai,and to explore the factors that affect their quality of life.Methods A cross-sectional study was conducted in Minhang District among HIV/AIDS patients.All subjects finished general situation questionnaire,Beck depression inventory (BDI),self rating anxiety scale (SAS),social support scale (SSS) and generic quality of life inventory-74 (GQOLI-74) survey.Results A total of 294 patients were recruited in this study with mean age of (39.6 ± 12.6) years old.Among the subjects,mean score of SAS was 40.5 ± 8.8,higher than national normative score (t =20.8,P＜0.001).The prevalence of anxiety was 13.9％.The mean score of BDI was 8.90 ± 8.59,and the prevalence of depression was 28.6％.The mean score of quality of life (QOL) was 66.6 ± 10.9.Multivariate logistic regression analysis showed that QOL of patients with high school or secondary school was lower than those with college and above education (OR =0.34,95 ％ CI:0.12-0.95).QOL scores of patients with moderate or severe depression were lower than those with less depressed patients,OR values were 0.14(95 ％ CI:0.06-0.34) and 0.07(95 CI:0.03-0.20)respectively.QOL of patients with anxiety was lower than the patients without anxiety (OR =0.10,95 ％CI:0.04-0.27).QOL of patients with high scores of social support was higher than the patients with low scores of social support (OR =3.95,95 ％ CI:1.82-8.59).Conclusions We should pay more attention to the psychological state of patients with HIV/AIDS.The quality of life can be improved by improving social support and reducing the occurrence of anxiety and depression.

Objective To understand the trends and epidemiological characteristics of syphilis in Minhang District of Shanghai,so as to provide scientific basis for making control strategies.Methods The data of reported cases of syphilis in Minhang District from 2005 to 2016 were analyzed with epidemiological methods.Results Overall 11 394 cases of syphilis were reported from 2005 to 2016,the incidence was 42.9 per 100 000 person-year.The incidences of Phase Ⅰ,Phase Ⅱ,Phase Ⅲ,genital and latent syphilis were 10.3,13.4,0.3,2.3 and 16.6 per 100 000 person-year,respectively.In 2009,the incidence reached 59.1 per 100 000 person-year and reach peak.The cases were constituted by local residents (62.4％) and residents from other provinces (37.6％),male patients predominated with male/female ratio of 1.1∶1.Most cases were aged from 25 to 54 years old(61.1 ％),and those people older than 54 years were more and more likely to be found infected.Local residents predominated by those who aged 25-64 years (74.2％) and other province predominated by 15-44 years (79.1％),the ratios of male/female were 1.3 ∶ 1 and 0.8 ∶ 1.Female were much younger than male,there were 50.0％ of female and 29.3％ of male who were aged between 15-34 years old.Latent syphilis was mostly reported in local male residents who were older than 55 years,female who were 25-54 years and other provinces' female aged 15-44.Phase Ⅰ and Ⅱ syphilis were mostly reported in local male residents less than 55 years old and other provinces' male less than 65 years old.There were significant difference between residence,sex,age and different kinds of syphilis with P＜0.001.Conclusions The trends of syphilis incidence was increasing before 2009 in Minhang District and then declining and slowing down in recent years.The measure of reinforcing monitor,propaganda and education,active screening syphilis in female and local old man should be taken to control and prevent the spread of syphilis.

OBJECTIVETo develop a primer-extension in combination with denaturing high-performance liquid chromatography (PE-DHPLC)-based assay for prenatal diagnosis of the five most common beta-thalassemia mutations in Chinese.

METHODSThe human beta-globin gene fragment was amplified by PCR, followed by a multiple PE reaction specific for each five mutations. Then the PE product mixtures were separated for genotyping of beta-globin gene mutations using fully-denaturing DHPLC analysis.

RESULTSIn a blind study, prenatal diagnosis was performed on thirty-six at-risk families for beta-thalassemia major. Reverse dot blot (RDB) analysis was used to validate each result, showing an accuracy rate of 100% for PE-DHPLC in a total of 108 samples tested. Overall, by PE-DHPLC analysis, the authors could identify the genotypes involving the five mutations and normal alleles corresponding to 94.4% (102/108) and actually make final decision for prenatal diagnosis covering 97.2% (35/36).

CONCLUSIONThe PE-DHPLC protocol can be a simple, rapid, and highly accurate assay in the prenatal detection of common beta-thalassemia mutations.

Base Sequence ; Chromatography, High Pressure Liquid ; methods ; DNA Mutational Analysis ; methods ; DNA Primers ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genotype ; Globins ; genetics ; Humans ; Molecular Sequence Data ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; beta-Thalassemia ; diagnosis ; genetics

OBJECTIVESTo study the interaction of hepatitis virus B (HBV) and tumor suppressor p53.

METHODSPlasmid pCMVp53 was transfected or cotransfected with pCMVHBVa (wild type HBV) or PCMVHBVb (mutant type HBV) into the hepatoma cell line SMMC-7721 by lipofectamine. Apoptosis cells were labeled by annexin V-FITC and confirmed by flow cytometry. Reporter plasmids PG13-CAT or p21-luc were cotransfected respectively in each group to indicate transactivation activity of p53 and it's effect on p21 promoter. Western blot was performed to observe p53 expression in each group.

RESULTSThe group transfected by pCMVp53 alone exhibit higher luciferase activity and higher apoptosis rate, otherwise, p53 expression, enzyme activity of PG13-CAT or p21- luc and cell apoptosis rate were much higher in the group cotransfected by pCMVp53 and pCMVHBVa, but not in the other cotransfected group; HBV replication was enhanced in p53 cotransfected group.

CONCLUSIONp53 expression and effects could be enhanced by HBV and p53 had positive regulation effect on HBV replication.

Apoptosis ; Carcinoma, Hepatocellular ; genetics ; pathology ; virology ; Cell Line, Tumor ; Hepatitis B virus ; genetics ; physiology ; Humans ; Liver Neoplasms ; genetics ; pathology ; virology ; Luciferases ; metabolism ; Promoter Regions, Genetic ; Proto-Oncogene Proteins p21(ras) ; genetics ; Trans-Activators ; genetics ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Virus Replication

OBJECTIVETo explore the biological impact of 40 amino acid deletion at the C-terminal of hepatitis B virus X on the proliferation of hepatoma cells.

METHODSCells of SMMC-7721 hepatoma cell line were transfected with HBx and its derivative HBx3'-40, harboring the 40 amino acid deletion at the distal C-terminal region. Cell growth curve, colony formation in soft agar plate and tumorigenesis assay in nude mice were used to observe the alterations induced by the transfection of HBx and HBx3'-40. The expression level of PCNA in tumor cells was also investigated.

RESULTSThe growth rates of the cells transfected with HBx and HBx3'-40 were markedly increased as compared with that of the control group. The colony formation rates were enhanced in the cells transfected with HBx(48.7 +/- 8.1) and HBx3'-40 (82.8+/-6.0), comparing with the control (26.9 +/- 3.5) %. In the tumorigenic assay, the size and weight of tumors were significantly increased in the cells transfected with HBx (0.412 +/- 0.212, 0.395 +/- 0.159) % and HBx3'-40 (1.476 +/- 0.232, 0.987 +/- 0.279) %, as compared with the control group (0.051 +/- 0.024, 0.033 +/-0.004) %. The expression level of PCNA in tumors was increased in both HBx (59.00 +/- 2.58) % and HBx3'-40 (69.25 +/- 3.77) % transfected cells, comparing with the control (37.67 +/- 2.52) %. Overall, the cells transfected with HBx3'-40 demonstrated the highest proliferative capacity.

CONCLUSIONThe deletion of 40 amino acids in the C-terminal of HBx is correlated with an enhanced proliferation of hepatoma cells and may play an important role in the malignant transformation of the liver.

Amino Acid Sequence ; Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; metabolism ; Sequence Deletion ; Trans-Activators ; genetics ; Transfection

OBJECTIVETo investigate the expression and significance of peroxisome proliferators-activated receptor gamma (PPAR gamma) in human glioma.

METHODSImmunohistochemical staining for PPAR gamma was performed using biopsy specimens of human glioma of various histological types. Expression of PPAR gamma and GFAP in glioma cell lines SWO-38, U251 and SHG-44 were analyzed using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSImmunohistochemical study showed that PPAR gamma was expressed in glioma tissues with positive rate of 37.5%. Western blotting and RT-PCR showed that PPAR gamma was expressed in both glioma cell lines SWO-38 and U251, but not in SHG-44 cells. However, high expression of GFAP was detected in SHG-44 cells.

CONCLUSIONPPAR gamma is associated with carcinogens of glioma. Actived PPAR gamma by agonist may be a novel approach to the treatment of glioma.

Blotting, Western ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Glial Fibrillary Acidic Protein ; biosynthesis ; genetics ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; PPAR gamma ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo evaluate the perinatal management and outcome of different types of fetal arrhythmia.

METHODSA retrospective analysis was conducted among the fetuses with arrhythmia identified by M-mode and pulsed Doppler echocardiography in a single institution between October 2003 and December 2010.

RESULTSA total of 130 fetuses were found to have fetal arrhythmia. The most common arrhythmia during pregnancy was extrasystole (n=59), followed by bradycardia (n=23), tachycardia (n=16), atrial flutter (AF, n=3), atrioventricular block (AVB, n=12) and other arrhythmia (n=17). The overall incidence of cardiac anomalies (commonly fetal bradycardia) was 9.2% in these cases. The prognosis of arrhythmia differed significantly between cases of different classifications. The type of fetal arrhythmia (P=0.024), presence of congenital heart defect (CHD, P=0.000) and fetal hydrops (P=0.008) were significant risk factors associated with termination of pregnancy.

CONCLUSIONFetal arrhythmias without CHD or hydrops under close monitoring often have good clinical outcome, while fetal bradycardia is associated with a high mortality rate. CHD and the presence of fetal hydrops are significant risk factors for pregnancy termination.

Adult ; Arrhythmias, Cardiac ; classification ; diagnostic imaging ; Female ; Fetal Diseases ; diagnostic imaging ; Heart Defects, Congenital ; diagnostic imaging ; Humans ; Pregnancy ; Pregnancy Trimester, Second ; Pregnancy Trimester, Third ; Retrospective Studies ; Ultrasonography, Doppler, Color ; Ultrasonography, Prenatal ; Young Adult