Objective To detect the mutations in RECQL4 gene in a Chinese patient with Rothmund- Thomson syndrome (RTS). Methods Blood samples were collected from a sporadic patient with RTS, his unaffected parents and 30 unrelated population-matched controls. DNA was extracted, and all the coding sequences of RECQL4 gene were amplified by PCR. Direct sequencing was performed with the amplicons to detect the possible mutations in these subjects. Results Two mutations, i.e., IVS11-1G > A and 3401 A >T, which resulted in a premature termination codon at amino acid 560, were found in the RECQL4 gene of the patient. His father was heterozygous for IVS11-1G > A, and his mother for 3401 A>T. Meanwhile, neither of the two mutations were observed in 30 unrelated normal control individuals. Conclusion Two mutations, including IVS11-1G>A and 3401 A>T are present in the RECQL4 gene of the sporadic patient with RTS.

Objective To establish a cell line with overexpression of rat serotonin1A receptor (5-HT1AR).Methods Human neuroblastoma cells-SH-SY5Y were donated by cancer institute attached to the 4 th Affiliated Hospital, Hebei Medical University. Total RNA was extracted from brain tissues of male SD rats and rat 5-HT1A R was obtained by RT-PCR. Plasmid pc-DNA3. 1/hisC containing the rat 5-HT1AR (pc-DNA3.1/hisC-Rat-5-HT1AR)was constructed and transfected into SH-SY5Y cells. The transfected cells were isolated by G418 selection and SH-SY5Y-Rat-5-HT1A R cells were obtained. Expression of 5-HT1A R was detected by Western blot analysis. Cell viability was evaluated by MTT assay. SH-SY5Y-Rat-5-HT1AR cells were further observed for 5-HT1AR by immuno-fluorescence staining. Results Plasmid pc-DNA3. 1/hisC-Rat-5-HT1AR was successfully constructed by linking Rat-5-HT1A R with pc-DNA3.1/hisC and transfected into SH-SY5Y. The SH-SY5Y-Rat-5-HT1A R cells were more slender than SH-SY5Y cells with less and longer processes. MTT showed that the viability of SH-SY5Y-Rat-5-HT1A R cells was much lower than SH-SY5Y. Rat 5-HT1A R was expressed efficiently on the membrane of SH-SY5Y-Rat-5-HT1A R cells. Conclusion A cell line with overexpress of rat 5-HT1A R is successfully established.

Objective To Compare different methods of quantitative breast density measurement.Methods The study included sixty patients who underwent both mammography and breast MRI. The breast density was computed automatically on digital mammograms with R2 workstation. Two experienced radiologists read the mammograms and assessed the breast density with Wolfe and ACR classification respectively. Fuzzy C-means clustering algorithm (FCM) was used to assess breast density on MRI. Each assessment method was repeated after 2 weeks. Spearman and Pearson correlations of inter- and intrareader and intermodality were computed for density estimates. Results Inter- and intrareader correlation of Wolfe classification were 0. 74 and 0. 65, and they were 0. 74 and 0. 82 for ACR classification respectively.Correlation between Wolfe and ACR classification was 0. 77. High interreader correlation of 0. 98 and intrareader correlation of 0. 96 was observed with MR FCM measurement. And the correlation between digital mammograms and MRI was high in the assessment of breast density (r = 0. 81, P ＜ 0. 01). Conclusion High correlation of breast density estimates on digital mammograms and MRI FCM suggested the former could be used as a simple and accurate method.

est that childhood wheezing is closely related to genetic factors, and children who carry predisposing genes may suffer from frequent wheezing and subsequent asthma.

AAIM:To investigate the effect of atorvastatin ( ATO) on electrical remodeling, atrial ion channel protein expression and cardiac function in atrial tachypacing rabbits, and to explore the potential electrical mechanism of ATO in the prevention of atrial fibrillation.METHODS:The rabbits were subjected to atrial tachypacing at 600 min-1 in the absence or presence of treatment with atorvastatin (ATP and ATO groups) for 48 h, and the other 10 as sham group without pacing ( NP group) .The tachypacing model was performed by attaching pacing and testing electrodes to left atrial and connecting with custom animal cardiac pacemaker in the open-chest situation.The animals in ATO group were pretrea-ted with ATO for 7 d and continued during tachypacing.Serial atrial effective refractory period ( AERP) was measured in each rabbit at baseline, 8 h, 16 h, 24 h, 32 h, 40 h and 48 h with different cycle lengths.The changes of cardiac func-tions and cardiac structure were observed by cardiac ultrasonic cardiogram before and after atrial tachypacing.The expres-sion of atrial ion channel proteins CaLα1 and Kv4.3 was detected by Western blotting.RESULTS:Compared with NP group, AERP at cycle lengths of 150 and 200 ms, the adaption of AERP, and the levels of CaLα1 and Kv4.3 expression were all decreased in ATP and ATO group, especially in ATP group.Left atrial dimension ( LAD) was increased in pacing groups as compared with NP group (P<0.05) after pacing delivery for 48 h, while no difference between the formers was observed.No significant change of the left ventricular dimension ( LVD) and ejection fraction ( LVEF) among groups be-fore and after pacing was found.CONCLUSION:Atrial tachypacing significantly shorten AERP, resulting in poor adap-tion of AERP, while ATO pretreatment significantly attenuates the atrial electrical remodeling in rabbits, but had no effect on cardiac structure.ATO suppresses the down-regulation of atrial ion channel proteins CaLα1 and Kv4.3 expression after 48 h, which may be the potential ionic mechanism of atrial electrical remodeling for ATO.

Objective To investigate the changes in the concentration of myocardin in children with lupus nephritis (LN) under different degree of vessel damage.Methods Forty-nine children diagnosed with LN by routine tissue immunolfuorescence, light microscope, and electron microscope were included, and 30 healthy children were included as control group. The pathological classiifcations were performed according to the ISN/RPS 2003 LN pathological classiifcation criterion. According to the Katafuchi evaluation method, the semi quantitative assessment of glomerular and kidney tubule damage was carried out, and the degree of vascular damage was evaluated at the same time. Double antibody sandwich method was used to detect the concentration of serum myocardin.Results The glomerular and kidney tubules damage in children with LN were signiifcantly aggravated with higher pathological classification (P<0.05). Glomerular damage was positively correlated with renal interstitial damage (r=0.96, P<0.01). The degree of vascular damage was related to the degree of glomerular injury and renal interstitial injury, while it was no related with the results of clinical tests. There were different concentrations of myocardin among mild-, moderate-, severe-vessel damage and control groups (F=378.61,P<0.001), and the concentration of myocardin in moderate- and severe-vessel damage groups were obviously lower than those in control group and mild-vessel damage group (P<0.01) while there was no difference between control group and mild-vessel damage group (P>0.05). According to pathological type, there were signiifcant differences in the concentration of myocardial between control group and different pathological types (F=626.793,P<0.01). FromⅡ,Ⅲ,Ⅲ+Ⅴ,Ⅳ toⅣ+Ⅴ, the concentrations of myocardial were decreased systematically, and there were statistic differences between groups (P all<0.05).Conclusion The concentration of myocardin in children with LN can relfect the renal vascular damage to a certain extent. Elevation of myocardin concentration may be helpful for the repair of vascular damage.

Objective To investigate the roles of mitochondrial DNA (mtDNA) in the respiratory function of cerebral mitochondria in rats exposed to acute hypoxia by observing the changes of mitochondrial respiratory function and cytochrome C oxidase (COX) activity. Methods The rat cerebral cortex mitochondria were isolated by centrifugation. Mitochondrial respiratory function and COX activity were measured by Clark oxygen electrode. Results ① Compared with the control group (C), hypoxia group (H) showed significantly elevated state 4 respiration (ST4), decreased state 3 respiration (ST3), and respiratory control rate (RCR). ② ST3 in group of treatment with chloramphenicol (CAP) plus hypoxia (MH) was significantly lower than that in Group C, while ST4 in Group MH was significantly lower than that in groups C and H. RCR in Group MH was lower than that in Group C, but higher than that in Group H. ③ COX activity in Group H was significantly lower than that in Group C. In Group MH, COX activity was higher than that in Group H, but was still lower than that in Group C. Conclusion The complete expression of mtDNA may play an important role in mitochondrial respiratory function. CAP treatment might be beneficial to the recovery of rat respiratory function.

Objective To observe the adipocytic differentiation potential of bone marrow stromal cells (BMS), and the effect of simvastatin on adipocytic differentiation of bone marrow stromal cells in vitro, and to elucidate the mechanisms of anabolic effect of simvastatin on bone formation. Methods BMS from femur and tibia of adult female BALB C mice were cultured in vitro. Changes of alkaline phosphatase (ALP) activity were determined after treatment with adipogenetic agonist (hydrocortisone 0 5 ?mol/L and indomethacin 60 ?mol/L, HI) for 6 days. Thenexpression of lipoprotein lipase (LPL) mRNA was detected by RT PCR after treatment with HI and different concentration of simvastatin for 72 h. Adipogenetic differentiation were also observed with Oil Red O staining and fluorescence activated cell sorting (FACS) after treatment with HI and different concentration of simvastatin or 100 ?g/L rhBMP 2 for 12 days. Results After BMS were treated with HI for 6 days, ALP activity was significantly decreased ( P

Objective: To observe the effect of simvastatin on osteoblastic cell differentiation of bone marrow stromal cells in vitro, and to elucidate the mechanisms of anabolic effect of simvastatin on bone formation. Methods: Bone marrow stromal cells from femur and tibia of adult female BALB C mice were cultured in vitro , after being treated with different concentrations of simvastatin for 72 h, changes of mRNA level of osteocalcin (OCN) were detected by RT PCR, change of OCN, and osteopontin (OPN) expression were examined by Western blot, and the changes of cellular alkaline phosphatase activity (ALP) were examined by histochemistry and enzymologic measurement. Results: After bone marrow stromal cells were treated with different concentration of simvastatin for 72 h, level of OCN mRNA increased, and expression of OCN and OPN also increased in a concentration dependent manner, and cellular ALP activity significantly increased in a concentration dependent manner. Conclusion: Simvastatin can stimulate osteoblastic differentiation,and improve cellular ALPase activity with high expression of osteocalcin and osteopontin in vitro. These may be parts of the mechanism of anabolic effect of simvastatin on bone formation.

Objective To assess whether the plaque volume of carotid artery evaluated by three-dimensional ultrasound can predict the risk of cardiovascular events earlier in patients with coronary heart disease, in comparison with plaque thickness.Methods A total of 99 patients were enrolled, including 66 patients with asymptomatic carotid plaque and 33 patients with coronary heart disease (CHD) and carotid plaque.Three-dimensional ultrasound was performed in all the patients.Coronary angiography was performed in all the patients in CHD group.Coronary artery Gensini score was calculated.Plaque thickness, plaque area and plaque volume were compared between the two groups.We analyzed the relationship of plaque volume with Gensini score and other cardiovascular risk factors.Results Plaque volume was significantly higher in CHD group [(1312.38±513.70)mm3] than in asymptomatic group [(947.54±321.41)mm3] (P=0.023).However, plaque thickness and plaque area were not significantly different between the two groups (P>0.05).Plaque volume was positively correlated with Gensini score (r=0.519, P=0.002), homocysteine (r=0.569, P=0.002), and cardiovascular risk factors such as diabetes, blood glucose and glycosylated hemoglobin.There was a negative correlation between plaque volume and high-density lipoprotein (HDL), a factor protecting against cardiovascular events (r=-0.387, P=0.038).Conclusion Compared with plaque thickness, the volume of carotid plaques assessed by three-dimensional imaging technique may be a better predictor of the risk of cardiovascular events in patients with coronary heart disease.