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

BACKGROUND: There are many in vitro selection method of sperm, and swim-up and density gradient centrifugation are commonly used. It remains unclear which method minimizes bad stimulation to the sperm and select sperm with high fertilization potential. OBJECTIVE: To evaluate the effectiveness of swim-up and gradient centrifugation preparation techniques on intrauterine insemination (IUI).METHODS: A computer-based online search of Cochrane Library, PubMed, EMBASE databases was performed, and some related journals were manually searched for related articles published between January 1966 and February 2009. The quality of included randomized controlled trials (RCT) and q-randomized trials (Q-RCT) was evaluated and Meta-analysis was conducted by the Cochrane Collaboration's software RevMan5.0. Experts.RESULTS AND CONCLUSION: A total of 6 studies were included, involving 4 RCTs and 2 Q-RCTs. A total of 486 patients (1 099 IUI cycles) were enrolled. The Meta-analysis indicated that there was no difference between swim-up and gradient centrifugation preparation techniques for the IUI in terms of cycle pregnancy rates [OR = 1.11, 95%CI(0.8,1.55)], miscarriage rates [OR = 0.31, 95%CI(0.09,1.04)], sperm count [the weight mean difference (WMD) =-0.89, 95%CI(-14.17,12.38)], sperm motility [WMD = -2.31, 95%CI(-7.27,2.65)]. There is insufficient evidence to confirm which is the best method in the two specific preparation techniques. The quality of study methods should be improved. And more measure parameters should be included when comparing it before or after treatment, such as sperm motility, sperm count, sperm function.

Adult ; Aspergillosis ; complications ; Female ; Hepatitis B, Chronic ; complications ; Hepatitis, Viral, Human ; complications ; Humans ; Lung Diseases, Fungal ; complications ; Male ; Middle Aged

OBJECTIVETo study the changes of body composition in females patients with human immunodeficiency virus (HIV)-related lipodystrophy (LD) syndrome (HIV-LD).

METHODSTotally 25 female patients who were treated in our hospital from January 2002 to December 2009 were divided into LD group and non-LD group based on the existence of LD. All these patients were receiving highly active antiretroviral therapy (HAART). In addition, 12 healthy women were set as the controls. Total and regional body composition were measured by dual X-ray absorptiometry in all three groups.

RESULTSThe fat mass (FM) was correlated negatively with the duration of HAART (r=-0.431, P=0.029). Multiple linear regression analysis showed that FM had positive correlation with weight and negative correlation with lean mass (LM) (r = - 0. 973, P =0. 000). Total, trunk and leg FM were significantly lower in LD patients than that in controls (P <0.05).Meanwhile, total, trunk and leg bone mineral contents were statistically lower in LD patients than that in controls (P <0. 05). Lumbar bone mineral density of LD patients was lower than that of non-LD patients and controls, and there was significant difference between LD patients and controls (P = 0. 001). LM of LD patients was higher than that of non-LD patients but without statistical difference (P > 0. 05).

CONCLUSIONSThe peripheral and central FM and bone mineral contents remarkably decrease in female patients with HIV-LD. How-ever, HIV-LD patients tend to have higher LM than non-LD patients. .

Adipose Tissue ; metabolism ; Adult ; Body Composition ; physiology ; Bone Density ; physiology ; Female ; HIV-Associated Lipodystrophy Syndrome ; metabolism ; Humans ; Middle Aged ; Young Adult

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