Female ; Hepatitis C ; complications ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Interferons ; therapeutic use ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; complications ; drug therapy ; virology ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Treatment Outcome

Objective In order to further study the influence of a mutant on viral replication and transfection, a eukaryotic vector with mutation of 20/21 bp deletion (1748/ 1747 to nt 1767) in core promoter region and precore stop mutation (nt.1896) was constructed. Methods A linearized genome containing the entire HBV 3.5kb mRNA transcriptional units (P3.8Ⅰ vector) and initiating from the basic core promoter upstream sequences was used as a tool, the objective eukaryotic vectors were constructed by the molecular cloning and PCR based site directed mutagenesis in vitro. The capability of progeny virus production and transcription were examined with Southern blot and Northern blot analysis respectively, after transfection of the recombinant HBV plasmids into HepG2 cells by using liposome. Results The eukaryotic vectors were constructed successfully and their sequences were confirmed by clone sequencing. Both Southern and Northern blotting of DNA and RNA extracted from the transfected cells showed markedly reduced mutant activity to produce progeny virus, to transcript both 3.5kb precore/pregenome mRNA and 2.1kb preS/S mRNA. Conclusions The levels of replication and transcription are markedly reduced in the mutant compared with those in wild type HBV.

To elucidate whether the mutations in X region of hepatitis B virus (HBV) might be responsible for the different clinical profiles in cases positive for antibody to hepatitis B e antigen. The nucleotide sequences of X gene regions in serum HBV were examined in 14 asymptomatic carriers (AsC) and 14 chronic active hepatitis (CAH) patients with antibody to hepatitis e antigen. The results showed that 12 of 14 AsCs (85.7%) had insertions, deletions or point mutations in nucleotide sequence of X region resulting in truncation of the X protein by creating frame shift mutation or a new stop codon, whereas no patient with CAH had those X gene mutations( P

Antiviral Agents ; therapeutic use ; Guanine ; analogs & derivatives ; therapeutic use ; Hepatitis B ; immunology ; Hepatitis B Antibodies ; immunology ; Hepatitis B Surface Antigens ; immunology ; Humans ; Reproducibility of Results ; Stem Cell Transplantation ; Stem Cells ; Tissue Donors

OBJECTIVE:To study the spermicidal and antibacterial effects of tannic acid from Chinese gall.METHODS:Human sperm samples of 20 subjects were taken for spermicidal tests in vitro in accordance with the standard method recommended by WTO.The minimum inhibitory concentration and minimum bactericidal concentration(MIC and MBC)of tannic acid from Chinese gall on bacterium coli and staphylococcus aureus were detected.RESULTS:The lowest effective spermicidal concentration of tannic acid at 20s was 20mg? mL-1.The MIC and MBC of tannic acid against bacterium coli(3 strains)were 0.195～ 0.390mg? mL-1 and 0.390～ 0.780 mg? mL-1 respectively,against staphylococcus aureus(2 strains)were 0.049～ 0.098mg? mL-1 and 0.195～ 0.390mg? mL-1 respectively.CONCLUSION:Tannic acid from Chinese gall showed potent coagulating effect on spermatines and remarkable antibacterial effect,and which is expected to be an effective and safe vaginal spermicidal and antibacterial agent,further study on which remains to be carried out.

Animal models of pulmonary artery hypertension (PAH),aiming to simulate human characteristics of the disease,have contributed extensively to understanding the pathophysiology of PAH and the investigation of experimental treatments.The classical models include monocrotaline models,chronic hypoxia model and so on,more new models were investigated in recent years.These animal models were not able to perfectly mimic human pathological characteristics of PAH because of the defect in different aspects.In this review,both typical and novel methods of PAH modeling were summarized and evaluated to provide a suitable guidance for the settlement of animal models which can meet human characteristics comprehensively.

Objective To investigate the effects of H3K9ac hyperacetylation mediated by histone acetylases on the overexpression of MEF2C in the hearts of fetal mice exposed to alcohol during pregnancy,and provide new interven-tion targets for prevention and treatment of cardiac dysplasia caused by alcohol exposure.Methods C57BL/6 mice were divided into 5 groups randomly,blank control group,dimethylsulfoxide (DMSO)group,alcohol group,alcohol +anacardic acid group and anacardic acid group,and then the hearts of fetal mice were collected to be analyzed.Chroma-tin immunoprecipitation and Western blot were used in assaying the binding of histone acetylases and the level of H3K9ac to the promoter of MEF2C in the hearts of fetal mice.The mRNA expression of MEF2C was tested by adopting real -time PCR.Results The level of H3K9ac in the promoter of MEF2C in the hearts of fetal mice exposed to alcohol was higher than that in the hearts of fetal mice exposed to saline (1 .30 ±0.1 9 vs 0.45 ±0.01 ),there was statistically significant difference (P <0.05),while the binding of E1 A -binding protein (p300),p300 /cyclic adenosine mono-phosphate response element binding protein -associated factor (PCAF)and steroid receptor coactivator -1 (SRC1 )to the promoter of MEF2C were abnormally elevated in the hearts of fetal mice treated by alcohol (1 .68 ±0.08 vs 0.82 ± 0.08,1 .08 ±0.05 vs 0.42 ±0.02,1 .1 8 ±0.05 vs 0.39 ±0.08),and there were statistically significant differences (all P <0.05).The expression of MEF2C mRNA in alcohol group was higher than that in blank control group (1 .36 ± 0.1 2 vs 0.29 ±0.03),there was statistically significant difference(P <0.05).However,a pan -acetylases inhibitor, anacardic acid,could decrease significantly the binding of p300 and PCAF to the promoter of MEF2C (1 .52 ±0.05 vs 0.63 ±0.09,1 .1 3 ±0.04 vs 0.45 ±0.04),and correct abnormal hyperacetylation of H3K9ac induced by alcohol (1 .58 ±0.08 vs 0.67 ±0.05),and down -regulate the over -expression of MEF2C in the hearts of fetal mice exposed to alcohol (1 .36 ±0.1 2 vs 0.41 ±0.05 ),and there were statistically significant differences (all P <0.05 ). Conclusions Hyperacetylation of H3K9ac mediated by p300 and PCAF may be a key regulatory factor in the over -expression of cardiac nuclear transcription factor MEF2C in the hearts of fetal mice exposed to alcohol during pregnan-cy.Anacardic acid can significantly attenuate the level of H3K9ac through inhibiting the binding of p300 and PCAF to the promoter of MEF2C,and down -regulate the over -expression of cardiac nuclear transcription factor MEF2C in the hearts of fetal mice.

Anthocyanins are a ubiquitous group of water-soluble plant pigments of the flavonoid family, with anticancer property through HER-2 signaling pathway. Nowadays, molecular docking plays an important role in exposing the active sites and obtaining the bioactive conformation involving protein-ligand interactions. According to the crystal structure of HER-2 kinase domain and 12 main antitumor compounds of anthocyanins as well as ATP, a molecular docking study was performed by MVD program. All 12 compounds could bind to the same cavity of HER-2 kinase domain by high affinity (MolDock Score < -105 kJ/mol for anthocyanidins, < -130 kJ/mol for anthocyanidins-glc), where hydrophobic force and hydrogen bond played key roles. Additionally, this cavity overlapped with ATP binding (MolDock Score = -161 kJ/mol) domain; the binding of anthocyanins presumably interfered the H bond formation between ATP and HER-2. These results indicate that anthocyanins may competitively bind to ATP binding site in HER-2 kinase domain by suppressing HER-2 activation and downstream signaling cascade. This may provide useful theoretical instruction for the molecular mechanism of HER-2 kinase activity inhibition by anthocyanins in cancer prevention and treatment.
Anthocyanins ; chemistry ; Catalytic Domain ; Hydrogen Bonding ; Hydrophobic and Hydrophilic Interactions ; Molecular Docking Simulation ; Phosphorylation ; Protein Interaction Domains and Motifs ; Receptor, ErbB-2 ; chemistry

Objective: To explore the dynamic regulation of histone acetylases p300 and p300/CBP associated factor (PCAF) on cardiac development gene NKX2.5 during cardio-genesis and to provide the new theoretical basis to clarify the regulatory mechanism for cardio-genesis in fetal mice.
Methods: Our research included 4 groups of cardiac tissues: Embryo (EB) 14.5 days group,n=10, EB 16.5 days group, n=10 and Neonatal 0.5 day group,n=5, Neonatal 7 days group,n=3. Immunoprecipitation was performed in myocardial tissues using anti-p300, anti-PCAF and anti-H3K9ac antibodies to retrieve p300, PCAF and H3K9ac binding DNA, the speciifc DNA sequences were ampliifed by real-time PCR to detect and the binding levels of p300, PCAF and the acetylation level of H3K9ac in NKX2.5 promoter sequence. In addition, the mRNA expression of NKX2.5 was examined by RT-PCR.
Results: The binding levels of p300 and PCAF had the timing consequence at different stage of cardio-genesis. The binding level of p300 in EB 16.5 days group (0.063 ± 0.021), Neonatal 0.5 day group (0.019 ± 0.008), Neonatal 7 days group (0.011 ± 0.003) were all lower than that in EB 14.5 days group (0.231 ± 0.033), and in Neonatal 0.5 day group and Neonatal 7 days group were lower than EB 16.5 days group, allP<0.05. The binding level of PCAF in EB 16.5 days group (0.063 ± 0.021), Neonatal 0.5 day group (0.019 ± 0.008), Neonatal 7 days group (0.011 ± 0.003) were all lower than that in EB 14.5 days group (0.185 ± 0.023), allP<0.05. The H3K9ac acetylation level and NKX2.5 mRNA expression level had the timing consequence at different stage of cardio-genesis. H3K9ac acetylation level in EB 16.5 days group (0.098 ± 0.014), Neonatal 0.5 day group (0.074 ± 0.010), Neonatal 7 days group (0.045 ± 0.014) were all lower than that in EB 14.5 days group (0.119 ± 0.020), and in Neonatal 7 days group was lower than EB 16.5 days group, allP<0.05. The NKX2.5 mRNA expression level in EB 16.5 days group (0.701 ± 0.181), Neonatal 0.5 day group (0.502 ± 0.159), Neonatal 7 days group (0.529 ± 0.13) were all lower than that in EB 14.5 days group (1.000 ± 0.130), allP<0.05.
Conclusion: Histone acetylases p300 and PCAF may dynamically regulate H3K9ac acetylation in NKX2.5 promoter sequence, and the mRNA of NKX2.5 was dynamically expressed during cardio-genesis in experimental fetal mice.

ObjectiveTo analyze the role of postmastectomy radiotherapy in different molecular subtypes of breast cancer patients with Stage T1 -T2 and one to three positive axillary nodes. MethodsA total of 436 breast cancer patients with T1 -T2 and one to three positive axillary lymph nodes treated with mastectomy and axillary dissection were retrospectively analyzed. Patients were grouped as the following four subtypes:Luminal A, Luminal B, Her2+ and triple-negative. The local recurrence (LR), distant metastasis ( DM ), disease free survival (DFS) and overall survival (OS) rates were compared between paitents with or without radiotherapy in univariate analyses. Multivariate analyses for LR were performed. Results The follow-up rate was 86. 0％. In patients with Luminal A subtype, radiotherapy decreased the 5-year LR rate (4.6％ vs 15.8％ ,x2 =5.74,P=0.017) but had no influences on DM, DFS or OS rates (17.2％ vs 19.7％,x2 =0. 17,P=0.682;77.0％ vs 67. 1％ ,x2 =1.99,P=0. 158 or87.4％:85. 5％ ,x2 =0. 12,P=0. 733 ). In patients with Luminal B subtype, radiotherapy decreased the 5-year LR rate (3.7％ vs 12. 1％,x2 =4. 13, P =0. 042), increased DFS and OS ( 84. 0％ vs 57.6％ ( x2 =14.61, P =0. 000) and 91.4％ vs 70. 7％ ( x2 =11.87, P =0. 001 ), but had no influence on DM ( 12. 3％ vs 22. 2％, x2 =2. 97, P =0. 085).In patients with Her2+ subtype, radiotherapy decreased the 5-year LR rate (5. 6％ vs 31.0％ ,x2 =4. 31,P=0. 035) , increased DFS (61. 1％ vs 13. 8％ ,x2 =11.44,P=0.001 ) ,but had no influence on DM and OS (27.8％ vs 41.4％, x2 =0. 89, P =0. 345 and 66. 7％ vs 48. 3％, x2 =1.52,P =0. 218 ). In patients with triple-negative subtype, radiotherapy had no influence in LR, DM, DFS or OS (8. 7％ vs 26. 1％ ,x2 =2.42,P=0.120;39.1％ vs47.8％,x2=0.35,P=0.552;52.2％ vs 26.1％ , x2 =3. 29, P =0. 070 or 65.2％ vs 56. 5％ ,x2 =0. 37 ,P =0. 546). Tumor size and radiotherapy were independent prognostic factors for LR rate in multivariate analyses ( x2 =4. 76, P =0. 029 and x2 =8.06, P =0. 005 ). ConclusionsFor patients with stage T1 -T2 and one to three positive axillary nodes, patients with all molecular subtypes except triple-negative can benefit from postmasteetomy radiotherapy.