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.

Objective To discuss the therapeutic options for treatment of subaxial cervical dislocation combined with facet locking. Methods There were 49 patients with cervical dislocations including 7 patients with dislocation at C3,4, 15 at C4,5, 14 at C5,6 and 13 at C6,7. Eleven patients were with old dislocation, with duration of dislocation ranging from 2 hours to 61 days. Neurologic status of the patients according to Frankel scale was graded A in 14 patients, grade B in nine, grade C in 10 and grade D in nine. All patients were treated surgically after closed reduction with skull traction. Results The successful reduction rate was 63% for fresh dislocation, with average improvement of 0.65 grade for spinal cord function. All bone grafts got fusion at four months after operation. Conclusion Therapeutic options are based on fresh or old dislocations, paraplegia or not, intervertebral disk injury severity, and reduction or not through traction for patients with lower cervical dislocations.

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.

BACKGROUND:Studies have demonstrated that tetrandrine has protection on acute spinal cord injury,but the specific mechanism remains poorly understood.OBJECTIVE:To study the protection of tetrandrine on rat acute spinal cord injury and to study its mechanism from apoptosis pathway.METHODS:A total of 100 rats were randomly divided into 4 groups.All rats were prepared for spinal cord injury models using modified Allen method except that in the sham-surgery group.Methylprednisolone and tetrandrine was injected into rats in the methylprednisolone and tetrandrine groups by tail intravenous injection prior to and at 24,48 hours after model preparation.The same volume of physiological saline was injected in the sham-surgery and model groups.Basso-BeatUe-Bresnahan(BBB score)was recorded at 8 hours,1,3,7 and 14 days after model preparation.The morphological changes of spinal cord injury sites were observed by hematoxylin-eosin staining and the expressions of bcl-2 and bax were determined by immunohistochemistry.RESULTS AND CONCLUSION:The BBB score of methylpradnisolone and tetrandrine groups were significantly higher than that model group at 7 and 14 days(P<0.05),but there were no significant difference between the methylprednisolone group and tetrandrine group(P>0.05).Hematoxylin-eosin staining showed that the spinal cord injured severely at 3-7 days,the injury degree in the methylpradnisolone group and tetrandrine group was slighter than that of the model group,with smaller bax expression and greater bcl-2 expression(P<0.01).The findings demonstrated that,tetrandrine is able to protect neurons from apoptosis and promote the nerve function recovery by inhibiting the expression of Bax and promoting the expression of Bcl-2.Its effect is not inferior to methylprednisolone.

ObjectiveTo explore the clinical outcome of one stage posteroanterior decompression and bone implant in the treatment of severe lower cervical spinal bony canal stenosis. Methods The study involved 29 patients with severe lower cervical spinal bony canal stenosis treated with one stage posteroanterior decompression and bone implant from April 2006 to March 2009. There were 11 patients with old fractures, seven with posterior longitudinal ligament ossification and 11 with cervical disc calcification. The course of disease ranged from 2 months to 3.2 years, average 1.4 years. The nerve function was rated as grade B in two patients, grade C in 19 and grade D in eight according to Frankel scale. The average Japanese Orthopaedic Association (JOA) score was 9.8. ResultsAll patients were followed up for 7-28 months (average 15.2 months), which showed bony fusion five months after operation, with fusion rate of 100％. The Frankel grade was increased for average 1.2 grades and the nervous symptoms alleviated remarkably. Mean postoperative JOA score was 13.8 and increased for mean 4.0, with mean amehoration rate of 55.6％. ConclusionsOne stage posteroanterior decompression and bone implant is a safe and effective method for treatment of lower cervical spinal bony canal stenosis, when the intraoperative electrophysiological monitoring can assure the operative safety.

This study aims to assess the expression of Klotho and insulin-like growth factor-1 (IGF-1) and the association between Klotho and IGF-1 in rats with dementia model. Thirty rats were randomly divided into three groups. Morris water maze was used to investigate the learning and memory functions, and enzyme linked immunosorbent assay was used to analyze the levels of Klotho and IGF-1. Klotho and IGF-1 levels in the model group were lower than those in other 2 groups. Morris water maze test showed that the model group had longer escape latency times and shorter step platform times compared to other groups. Line correlation model demonstrated that Klotho level was positively correlated with IGF-1 level in rats with dementia (P= 0. 029). The levels of Klotho and IGF-1 both reduced at hippocampus in rats with dementia model, suggesting that it may be a close relationship between Klotho and IGF-1 in the pathogenesis of dementia.
Animals ; Dementia ; metabolism ; Female ; Glucuronidase ; genetics ; metabolism ; Hippocampus ; metabolism ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Male ; Maze Learning ; Memory ; physiology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effectiveness and the safety of clinical use of rhGH in the management of severe burn patients.

METHODSTwo hundred burn patients aged 20 - 25 yrs with TBSA from 20% to 50% and admitted within 5 postburn days (PBD) were enrolled in this study. The patients were divided into operation and non-operation groups. Furthermore, each group was randomly divided into control (N), treatment with rhGH in dosage 0.2 IU/Kg/d (A) and treatment with rhGH in dosage 0.4 IU/kg/d (B) groups. The general data, the metabolism of protein and glucose, the immune function, the urine biochemistry, the wound healing rate and hospital stay days recorded and compared among all groups.

RESULTSPlasma protein was increased, the immune function recovered early, the wound healing time was shortened obviously, the wound healing rate was increased evidently and the hospital stay days were decreased significantly in the treatment groups (A and B groups) compared with the N group. The body weight of burn patients remained unchanged in N group. Nevertheless, the plasma levels of glucose and insulin were increased and the urine output of potassium, sodium and chloride decreased in N group compared with those in A and B groups. Moreover, the levels of all these indices in B group were higher than those in A group, whereas side effects were more common in B group than in A group. In addition, the incidence of adverse reaction was higher in operation group than that in non-operation group.

CONCLUSIONrhGH could effectively promote protein synthesis, enhance systemic immune function, accelerate wound healing and shorten hospital stay days in severely burned patients. But attention should be paid to appropriate dosage and duration of use.

Adult ; Burns ; drug therapy ; immunology ; CD4-CD8 Ratio ; Dose-Response Relationship, Drug ; Growth Hormone ; therapeutic use ; Humans ; Middle Aged ; Wound Healing

The aim of this study was to investigate the expression of wt1 gene and the changes of gene expression in minimal residual disease (MRD) models (K562, HL-60 cell lines) and acute leukemia (AL) patients through inhibiting the expression of wt1 gene by antisense oligonucleotides (ASO). The bone marrow (BM) of 56 AL patients with complete remission (CR) was collected, then the BM samples with positive expression of wt1 gene were screened by RT-PCR. The cells of MRD model and screened wt1 gene positive samples were cultured and treated by ASO, then the changes of wt1 gene expression were detected. The results indicated that the sensitivity of wt1 gene was 10(-3)-10(-4), and the positive rate of BM wt1 gene expression in 56 AL patients with CR was 16%. After BM of 9 AL CR patients with MRD and MRD model (K562, HL-60 cells) expressing wt1 gene were treated by ASO, it was found that the wt1 expression in ASO group was blocked, while wt1 gene could be still detected in both sense oligonucleotides (SO) and control groups. It is concluded that ASO can obstruct the expression of wt1 gene on the residual leukemia cells in vitro.
Gene Expression ; HL-60 Cells ; Humans ; K562 Cells ; Neoplasm, Residual ; genetics ; Oligonucleotides, Antisense ; genetics ; WT1 Proteins ; genetics

OBJECTIVETo investigate the effect of ischemic preconditioning on chaperone hsp70 expression and protein aggregation in the CA1 neurons of rats, and to further explore its potential neuroprotective mechanism.

METHODSTwo-vesseloccluded transient global ischemia rat model was used. The rats were divided into sublethal 3-min ischemia group, lethal 10-min ischemia group and ischemic preconditioning group. Neuronal death in the CA1 region was observed by hematoxylineosin staining, and number of live neurons was assessed by cell counting under a light microscope. Immunochemistry and laser scanning confocal microscopy were used to observe the distribution of chaperone hsp70 in the CA1 neurons. Differential centrifuge was used to isolate cytosol, nucleus and protein aggregates fractions. Western blot was used to analyze the quantitative alterations of protein aggregates and inducible chaperone hsp70 in cellular fractions and in protein aggregates under different ischemic conditions.

RESULTSHistological examination showed that ischemic preconditioning significantly reduced delayed neuronal death in the hippocampus CA1 region (P < 0.01 vs 10-min ischemia group). Sublethal ischemic preconditioning induced chaperone hsp70 expression in the CA1 neurons after 24 h reperfusion following 10-min ischemia. Induced-hsp70 combined with the abnormal proteins produced during the secondary lethal 10-min ischemia and inhibited the formation of cytotoxic protein aggregates (P < 0.01 vs 10-min ischemia group).

CONCLUSIONIschemic preconditioning induced chaperone hsp70 expression and inhibited protein aggregates formation in the CA1 neurons when suffered secondary lethal ischemia, which may protect neurons from death.

Animals ; Brain Ischemia ; pathology ; Cell Count ; methods ; Cell Death ; Disease Models, Animal ; Gene Expression Regulation ; physiology ; HSP70 Heat-Shock Proteins ; metabolism ; Hippocampus ; blood supply ; metabolism ; pathology ; Ischemic Preconditioning ; Male ; Neurons ; metabolism ; Proteins ; metabolism ; Rats ; Rats, Wistar ; Time Factors

OBJECTIVETo investigate the effect of bufalin on nucleus-mitochondria localization of human telomerase reverse transcriptase(hTERT) by exploring its effect on proliferation and apoptosis in human esophageal squamous carcinoma EC9706 cells.

METHODSEC9706 cells were treated with bufalin at various concentrations, and then the cell growth inhibition of EC9706 cells was examined by CCK-8 assay and the 50% inhibitory concentration (IC(50)) was calculated.Cell cycle analysis was performed by flow cytometry with PI staining, and nucleus morphology of apoptosis were observed by fluorescence microscopy with Hoechst 33342 staining. The apoptotic index was measured by flow cytometry with Annexin V-FITC/PI double staining. hTERT subcellular localization and protein expression were determined by Western blotting and multiple immunofluorescence labling combined with laser confocal scanning microscopy.

RESULTSThe proliferation of EC 9706 cells was significantly inhibited by bufalin along with the increase of processing time and concentrations (p<0.01). After the EC9706 cells were exposed to 100 nmol/L bufalin,the number of cells gradually decreased in G(1) phase and increased in S and G(2)/M phases(p<0.05). The typical nucleus morphological changes of apoptosis were observed and the apoptotic index was increased(p<0.01). The expression of hTERT decreased in nucleus but increased in mitochondria(p<0.05).

CONCLUSIONSBufalin can inhibit the proliferation of human esophageal squamous carcinoma EC9706 cells in a time- and dose-dependent manner. It can arrest cell cycle in S and G(2)/M phases and induce the apoptosis of EC 9706 cells. hTERT is localized in both nucleus and mitochondria,and can be partially translocated from nucleus to mitochondria during the bufalin-induced apoptosis.

Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Telomerase ; metabolism