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.

Hepatitis B virus (HBV)-induced hepatocellular carcinoma (HCC) is one of the most frequently occurring cancers. Hepadnaviral DNA integrations are considered to be essential agents which can promote the process of the hepatocarcinogenesis. More and more researches were designed to find the relationship of the two. In this study, we investigated whether HBV DNA integration occurred at sites of DNA double-strand breaks (DSBs), one of the most detrimental DNA damage. An 18-bp I-SceI homing endonuclease recognition site was introduced into the DNA of HepG2 cell line by stable DNA transfection, then cells were incubated in patients' serum with high HBV DNA copies and at the same time, DSBs were induced by transient expression of I-SceI after transfection of an I-SceI expression vector. By using nest PCR, the viral DNA was detected at the sites of the break. It appeared that integration occurred between part of HBV x gene and the I-SceI induced breaks. The results suggested that DSBs, as the DNA damages, may serve as potential targets for hepadnaviral DNA insertion and the integrants would lead to widespread host genome changes necessarily. It provided a new site to investigate the integration.

Six 89bp primers were designed on the base of the cDNA sequence encoding the human leptin reported on the NCBI. The synthetic gene with 464bp encoding rhLep was obtained by SOE ( splicing by overlap extension) PCR. The expression vector pET22b(+)/rhLep was constructed and transformed into E. coli BL21 (DE3). The rhLep protein was expressed as inclusion bodies with the yield of more than 50% of total bacterial proteins after IPTG induction. The rhLep protein, which has a molecular weight about 16kD, was purified by Ni2+ affinity chromatography column and identified by SDS-PAGE. The MTT Assay shows that rhLep promotes EC304 cells growth at the low concentration of 10ng/mL to 30 ng/mL, and rhLep appears cytotoxic to EC304 cells with the high dose of 50ng/mL to 225ng/mL. The viability of EC304 cells decreases to 1.2% with the concentration of 225ng/mL of rhLep. The massive apoptosis of rhLep on EC304 cells is observed by AO-staining under fluorescent microscope. All these results would lay the foundation for the further study of its biological functions in vitro and in vivo.
Apoptosis ; drug effects ; Escherichia coli ; genetics ; Humans ; Leptin ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology

OBJECTIVETo compare the effect of the two methods of back propagation network (BPN) test on TCM syndrome typing of depression.

METHODSTest was carried out by two methods as following: (1) Cross train-test method: 1731 patients with depression typed to 5 syndrome types were randomly divided into 2 groups, and they were trained and tested in turn; (2) Round-Robin method: Test was conducted in an altered cycle mode, that is, in a cycle, one out of the 1731 patients were selected to be tested, while the others were trained, the next cycle started when the test on the selected patient was finished and another one for test was selected. In this way, one cycle after the other, until all patients had been tested.

RESULTThe total training sensitivity of the two methods was 97.9% and 98.2% respectively, and the total testing sensitivity was 72.7% and 74.2% respectively.

CONCLUSION(1) The five TCM syndrome types of depression could be well differentiated by BPN, which is valuable for TCM syndrome typing in certain extent; (2) The sensitivity of Round-Robin method is slightly higher than that of Cross train-test method, but in comparison between them no remarkable significance was shown.

Adolescent ; Adult ; Depressive Disorder ; diagnosis ; Diagnosis, Differential ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Neural Networks (Computer) ; Sensitivity and Specificity

OBJECTIVEImpact of the presence of bacteria associated with a marine dinoflagellate, Alexandrium tamarense CI01, on the growth and toxin production of the algae in batch culture was investigated.

METHODSPronounced changes in the activities of the algal culture were observed when the culture was treated with different doses of a mixture of penicillin and streptomycin.

RESULTSIn the presence of antibiotics at the initial concentration of 100 u/mL in culture medium, both algal growth and toxin yield increased markedly. When the concentration of antibiotics was increased to 500 u/mL, the microalgal growth was inhibited, but resumed in a few days to eventually reach the same level of growth and toxin production as at the lower dose of the antibiotics. When the antibiotics were present at a concentration of 1 000 u/mL, the algal growth was inhibited permanently.

CONCLUSIONSThe results indicate that antibiotics can enhance algal growth and toxin production not only through their inhibition of the growth and hence competition for nutrients, but also through their effects on the physiology of the algae.

Animals ; Anti-Bacterial Agents ; pharmacology ; Bacteria ; Dinoflagellida ; microbiology ; pathogenicity ; Eutrophication ; Marine Toxins ; biosynthesis ; Penicillins ; pharmacology ; Saxitoxin ; Streptomycin ; pharmacology

PURPOSE: To report the 5-year follow-up results of a randomized controlled trial comparing bipolar transurethral resection of the prostate (TURP) with standard monopolar TURP for the treatment of benign prostatic obstruction (BPO). MATERIALS AND METHODS: A total of 220 patients were randomized to bipolar plasmakinetic TURP (PK-TURP) or monopolar TURP (M-TURP). Catheterization time was the primary endpoint of this study. Secondary outcomes included operation time, hospital stay, as well as decline in postoperative serum sodium and hemoglobin levels. All patients were assessed preoperatively and followed-up at 1, 6, 12, 24, 36, 48, and 60 months postoperatively. Parameters assessed included quality of life, transrectal ultrasound, serum prostate-specific antigen level, postvoid residual urine volume, maximum urinary flow rates (Qmax), and International Prostate Symptom Score. Patient baseline characteristics, perioperative data including complications, and postoperative outcomes were compared. Complication occurrence was graded according to the modified Clavien classification system. RESULTS: PK-TURP was significantly superior to M-TURP in terms of operation time, intraoperative irrigation volume, resected tissue weight, decreases in hemoglobin and sodium, postoperative irrigation volume and time, catheterization time, and hospital stay. At 5 years postoperatively, efficacy was comparable between arms. No differences were detected in safety outcomes except that the clot retention rate was significantly greater after M-TURP. CONCLUSION: Our results indicate that PK-TURP is equally as effective in the treatment of BPO, but has a more favorable safety profile in comparison to M-TURP. The clinical efficacy of PK-TURP is long-lasting and comparable with M-TURP.
Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostate/*surgery ; Transurethral Resection of Prostate/adverse effects/*methods ; Treatment Outcome

OBJECTIVETo screen and identify differentially expressed genes in the hippocampus of the offsprings of lead exposed female rats in order to provide a theoretical basis for identifying learning and memory deficits related genes.

METHODSRNA was extracted from the hippocampus of young rats with learning and memory deficits due to maternal lead exposure. Suppression subtractive hybridization was used to identify the differentially expressed genes in the hippocampus.

RESULTSAn effective subtracted library was constructed which consisted of approximately 200 clones. Sequencing for the library identified 93 clones harboring insertion fragments which included 43 different genes and 4 unknown genes. These genes might be related to learning and memory deficits due to maternal lead exposure.

CONCLUSIONSThe up-regulated genes in the hippocampus of young rats from pregnant rats under lead exposure include some housekeeping genes and some proteins involved in cellular protein folding, signal transduction, stress response and DNA methylation. These proteins might be directly related to a significant reduction in learning and memory abilities in the young rats.

Animals ; Female ; Gene Expression Profiling ; Gene Library ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Learning ; drug effects ; Memory ; drug effects ; Nucleic Acid Hybridization ; methods ; Polymerase Chain Reaction ; Pregnancy ; Rats

BACKGROUNDGap junction channels formed by connexin43 (Cx43) protein are important in cardiac morphogenesis, and Cx43 gene is thought to be associated with congenital heart malformation (CHM). This study was undertaken to detect the mutations of Cx43 in fetuses with CHM.

METHODSCx43 extron DNA was amplified by PCR from 16 fetuses with a variety of CHM. The PCR products were analyzed by SSCP and DNA sequencing. Thirty children who had no CHM were selected as controls.

RESULTSEight homozygous mutations of Cx43 were observed in a fetus with double outlet right ventricule (DORV), five of the 8 mutations were missense mutations including Arg239Trp, Ser251Thr, Ala253Pro, Pro283Leu and Thr290Asn, and the remaining 3 were silent polymorphisms including Gly252Gly, Pro256Pro and Thr275Thr. No mutations were found in other fetuses and the control group.

CONCLUSIONSMutations of Cx43 may be associated with congenital conotruncal anomalies. PCR-SSCP is an effective method for screening the mutations of Cx43.

Connexin 43 ; genetics ; Fetus ; metabolism ; Heart Defects, Congenital ; genetics ; Humans ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA

Acute Disease ; Adolescent ; Diagnosis, Differential ; Female ; Hepatitis ; diagnosis ; Hepatitis, Autoimmune ; diagnosis ; Humans

Two plasmid constructs, pcE2 and pcE3, containing 3' fragment of open reading frame 2 (ORF2,1163 bp) of hepatitis E virus (HEV) and full-length ORF3 (369 bp), were injected into bilateral tibialis of Swiss mice respectively，for three times (0, 2nd and 4th weeks) and observed the HEV IgG by ELISA. HEV IgG was induced after the injection of pcE2 or pcE3 or both, and the percentage of seraconversion was 100% after two weeks of the third injection. Compared with injection of either construct, the antibody titers were higher in the group with combined injection of two constructs.