Objective To investigate the application of the Doppler echocardiography on the clinical value on the left ventricular myocardial remodel and dysfunction in neonates with hypoxia-induced pulmonary artery hypertension. Methods The Doppler echocardiography (ECHO) datas of 24 neonates of pulmonary diseases with secondary pulmonary artery hypertension were identified by color Doppler echocardiography,and compared with 14 neonates with pulmonary diseases without pulmonary artery hypertension. Results Pulmonary artery systolic pressrure (PASP) (38.23?3.26) mm Hg increased ,the diameter of right ventricular(11.92?2.18)mm vs (7.34?0.93) mm and pulmonary artery(10.20?1.21)mm vs (6.89?0.22) mm and diameter of left ventricular end-diastolic(19.74?0.71)mm vs (14.42?0.32) mm in PAH groups increased significantly ( P 0.05). There was positive correlation between the increased pulmonary artery pressure in pulmonary artery hypertension groups and the AV/EV of the mitral value orifice blood stream ( r=0.4126,P

OBJECTIVE:To study the antioxidant effects of Scutellaria baicalensis(SBE)on the model mice with Alzheimer’s disease. METHODS:All mice were treated by D-gal (150 mg/kg) and NaNO2 (100 mg/kg) for 60 d to reproduce the AD model mice,ip,once a day. Totally 60 mice were randomly divided into normal control group(isovolumetric sodium chloride injection), model group (isovolumetric sodium chloride injection),estradiol (0.4 mg/kg) group,and SBE high,medium and low doses (500,250 and 125 mg/kg). The mice were intragastrically administrated while modeling except normal control group,once a day, for continuous 60 d. The superoxide dismutase(SOD),catalase(CAT)activities and malondialdehyde(MDA)content in brain tis-sue ,and glutathione peroxidase (GSH-Px) activity in serum of mice were determined. RESULTS:Compared with normal control group,the SOD,CAT activities in brain tissue,and GSH-Px activity in serum of mice in model group were decreased,MDA con-tent in brain tissue were increased,with significant difference(P<0.01 or P<0.05);compared with model group,the SOD,CAT activities in brain tissue and GSH-Px activity in serum of mice in SBE high and medium dose groups were increased,MDA content was decrease,with significant difference (P<0.01 or P<0.05). CONCLUSIONS:SBE can enhance the antioxidant capacity in AD model mice by a mechanism that may be related to the improvement of related oxidation indicators’levels.

BACKGROUND: Recently,the technique of isolation procedure of viable islets cell from large animals has been mature and used in clinical islet cell transplantation. Some study results indicate that low yields of pig islet cell isolations is probably not due to the presence of higher or more aggressive enzyme activities during the porcine isolation procedure. Therefore, the causative factors resulting in the inconsistent results should be sought for the intrinsic properties of the pancreatic islet and pancreas. The purpose of the study is to get more pancreatic islet cells for diabetic patients.OBJECTIVE: To investigate the effect of single enzymatic solution (SES)and multi-enzymatic solution(MES) on adult porcine pancreatic islet cell isolation,and to provide the theory base for diabetic patients rehabilitation.DESIGN: A completely randomized and controlled experiment based on animals.SETTING: General surgery and gerontology department in a university hospital.MATERIALS: The experiment was conducted in the Second Affiliated Hospital of Harbin Medical University from June 2003 to May 2004. Twenty consecutive random-bred adult pigs, 12 - 24 months old, weighing average 100 kg,were selected for experiment from Hada Slaughterhouse of Harbin.INTERVENTIONS: After slaughtering,the spleentic lobe of the pancreas was excised using sterile surgical gloves and surgical instruments. Each was immediately transported in 500 mL of sterile RPMI1640 solution at 4 ℃ to the laboratory for processing. After removal of fat peritoneum and superficial blood vessel,each pancreas was first immersed in 1:5 000 Liquor chlorhexidine for 3 minutes,then washed three times with cold RPMI1640, next mechanically minced into 1 mm×1 mm×1 mm fragments,weighed,and equally divided into two groups. Adult porcine islets were isolated with two different collagenase solutions(SES and MES).MAIN OUTCOME MEASURES: Islet count was performed with DTZ-staining. The viability was assessed by trypan-blue staining. Porcine islet insulin-secretory function was assessed by insulin content of cultured porcine islets and by insulin release. Islet morphological integrity was finally established by electron microscope examination.RESULTS: Overall islet cell content were statistically different between the two methods[(1 782 ±427) IE/g vs (1 293 ±451) IE/g,P<0.05]. No significant difference was found in viability,function or morphology of islet between MES and SES(P>0. 05).CONCLUSION: An average of isletisolated with MES method represents a uniquely massive yield in comparison with that by SES method. A good viability was also confirmed by static incubation and culture. The transplantation of those pancreatic islet cells can help to control the syndromes of diabetes.

OBJECTIVE To investigate the role of XIAP in cisplatin-induced apoptosis and its possible mechanism. METHODS Hep-2 cells were treated with different concentrations of cisplatin for different times. Using crystal violet assay, RT-PCR and flow cytometry (FCM), we investingated the rate of the cell apoptosis and the expression of XIAP protein and mRNA at different times after treating with different concentrations of cisplatin. RESULTS Hep-2 cells were adherent and in normal survival condition with very low apoptosis rate. After treating with cisplatin, the rate of inhibition, the expression of XIAP protein and the rate of apoptosis were remarkably different between different concentrations and times. The expression of XIAP protein was down regulated accompanied by the augmentation of the rate of Hep-2 cell apoptosis. The products of RT-PCR were analyzed, demonstrating that the XIAP mRNA was down regulated with the increased concentrations of cisplatin overtime. Correlation analysis showed the rate of inhibition and the rate of apoptosis were positively correlated with increased concentrations of cisplatin for different times, however, the expression of XIAP protein was negatively correlated. The expression of XIAP protein and the rate of apoptosis were conversely correlated. CONCLUSION The most important pathway that cisplatin induces cell death is apoptosis. The levels of expression of XIAP proteinand mRNA in Hep2 cells are time-and concentration-dependent after treating with cisplatin. Down-regulation of the XIAP protein expression to augment the rate of apoptosis is one of the mechanisms for the cisplatin tokill the carcinoma cells.

Objective To investigate the accuracy of functional magnetic resonance imaging(fMRI) and contrast-enhanced ultrasound on evaluation of the breast cancer patients after neoadjuvant chemotherapy.Methods Forty-eight patients with breast cancer who were confirmed by biopsy,undergoing 4 cycles weekly dose-concentrated programs (wPC) neoadjuvant chemotherapy,evaluated the efficacy by fMRI and contrastenhanced ultrasound.Results In pathology,complete remission was 12 cases (25.0％,12/48),non remission was 13 cases (27.1％,13/48).In contrast-enhanced ultrasound,complete remission was 20 cases (41.7％,20/48),non remission was 10 cases (20.8％,10/48).In fMRI,complete remission was 17 cases (35.4％,17/48),non remission was 11 cases (22.9％,11/48).The P values more than 0.05 showed no statistically significant among three methods,the Kappa test showed that fMRI were superior to contrastenhanced ultrasound on evaluating the efficacy of neoadjuvant chemotherapy.Conclusions fMRI and contrast-enhanced ultrasound can be used in evaluating the efficacy of neoadjuvant chemotherapy,however,the data shows fMRI is superior to contrast-enhanced ultrasound.Contrast-enhanced ultrasound must be as the supplementary method.

BACKGROUND:Bone graft for acetabular reconstruction includes morselized bone graft, structural bone graft and hybrid bone graft, and the morselized bone has been widely used because of the advantages of simple
production and short healing time.
OBJECTIVE:To explore the key technologies and clinical effect of impaction bone grafting with morselized bone in total hip revision for AAOS Ⅲ acetabular deficiency.
METHODS:Sixteen cases of AAOS Ⅲ acetabular deficiency were treated with impaction bone grafting with morselized bone combined with metal devicesor constructive bone grafting. The hip Harris scores and
radiographic data were compared before and after treatment. The effect of impaction bone grafting with morselized bone on acetabular deficiency was assessed.
RESULTS AND CONCLUSION:Al the patients were fol owed-up at 3, 6, 12 months after surgery and every half a year successively. The pain of hip joints after operation was relieved significantly and the walking function was
restored. The hip Harris score was improved from 48.00 points before surgery to 84.94 points after surgery (P<0.01). Five cases were graded as excel ent, eight cases as good, two cases as average, and one case as poor. The excel ent and good rate was 81%, and the satisfying rate of the patients was 94%. The post-operative X-ray films of al the 16 patients showed that the acetabular rotation centers were recovered (near) to normal and the acetabular cups were covered wel by bone. The grafting bone particles got radiological osseointegration and the
acetabular cup prosthesis did not displaced, and no displacement and breakage happened to the metal devices.
Impaction bone grafting with morselized bone in total hip revision for AAOS Ⅲ acetabular deficiency can effectively reconstruct the acetabular bone structure, retain and restore the acetabular bone mass, and it has good technical advantages and good clinical effects.

Objective To set up a new diagnostic platform based on microarray exon-capture and next-generation sequencing for detecting small mutations in dystrophin gene.The sensitivity and specificity of the method were assessed in clinical settings and the distribution of small mutations in Chinese Duchenne muscular dystrophy/Becker muscular dystrophy (DMD/BMD) patients were also analyzed.Methods Forty-one DMD/BMD patients diagnosed by the clinical criteria without large deletion or duplication (≥ 1exon) were recruited from Peking Union Medical College Hospital consecutively.Genomic DNA was extracted from blood samples.The libraries were prepared.Then exon and intron-exon flanking sequences of DMD gene were captured by custom microarray.Targeted next-generation sequencing and Sanger Sequencing were conducted.The patients who were not detected any disease-causing mutation were performed muscle biopsy.Results Thirty-eight subjects were detected small mutations in DMD gene.All single nucleotide variants (SNVs) and insertion & deletions (INDELs) were validated by Sanger sequencing.Twenty-one novel mutations were reported.The distribution of SNVs and INDELs was similar to other international DMD databases.Upon immunohistochemistry staining of dystrophin protein,1 of 3 mutation-undetected patients was diagnosed as DMD,2 of them were excluded.The specificity of the method was 100％,while the sensitivity was 97.4％.Conclusions Our microarray-captured next-generation sequencing assay could detect SNVs and INDELs with high sensitivity and specificity.Its advantages are economic,time-saving and stable.The platform is suitable for clinical gene diagnosis.

Objective To investigate the mechanism of tumor necrosis factor-α (TNF)-α inhibiting osteo blastdifferentiation of mesenchymal stem cells (BMMSCs) in the pathogenesis of osteoporosis in the mouse model of systemic lupus erythematosus (MRL/lpr). Methods The femurs of MRL / lpr and C3He/HeJ mice were isolated, the bone structure were examined by hematoxylin-eosin (HE) staining. The proteins of TNF-α, NF-κB P50, bone morphogenetic protein -2 (BMP-2) and PSmad1/5/8 were measured by immunohistochemical stain. Bone marrow mesenchymal stem cells (BMMSCs) were isolated. After BMMSCs grew on the cover slips, the proteins on top of it were evaluated by immunohistochemistry stain. Moreover, the alkaline phosphatase (ALP) staining was employed for the measurement of the early osteogenic differentiation. BMMSCs together with hydroxyapatite were embedded subcutaneously in the nude mice and eight weeks later, the ectopic bone formation was evaluated. The recombinant human tumor necrosis factor receptor type Ⅱantibody fusion protein (etanercept) or normal saline was subcutaneous injected to the mice with lupus. After four weeks, the expression of these proteins was observed and the ectopic bone formation was investigated. Image-Pro plus 6.0 software was employed for imagine analysis, and Studentˊs t-test was used to test the differences between 2 independent groups. Results MRL/lpr mice showed decreased volume of cortex and the percentage of cortex to the volume of bone of MRL/lpr mice was significantly lower compared to control groups and with C3He/HeJ mice (13.96±0.25 vs 23.61±0.71, n=3, P<0.01). The protein levels of both TNF-αand NF-κB P50 on the femur of MRL/lprl mice were higher than those of the control group (0.643±0.051 vs 0.405±0.022, 0.917±0.023 vs 0.650±0.032, n=3, P<0.01). The expressions of BMP-2 on the femur of MRL/lpr mice were lower than those of the C3He/HeJ mice (0.52 ±0.03 vs 0.72 ±0.03, n=3, P<0.01). There was no difference in the expression of PSmad1/5/8 on the femur between the two groups by immunohistochemistry detection (1.264 ±0.021 vs 1.301± 0.044, n=3, P>0.05). The expressions of TNF-α and NF-κB P50 in BMMSCs of MRL/lprl mice were higher than those of the C3He/HeJ (0.184±0.021 vs 0.136±0.013, 0.132±0.021 vs 0.097± 0.014, n=3, P<0.01), while BMP-2 and PSmad were lower than those of the control group (0.128±0.013 vs 0.216±0.221, 0.115±0.023 vs 0.196±0.034, n=3, P<0.01). After 7 days of BMP-2 stimulation, the activities of ALP of BMMSCs from MRL/lprl mice were reduced detected by ALP staining and the osteoblast differentiation of these cells were decreased than BMMSCs from the control mice by HE and Masson staining. The percentage of the cortex to the volume of bone of the etanercept injection MRL/lpr mice was higher than that of the control group (21.8±1.0 vs 14.3 ±0.6, n=3, P<0.01). Moreover, the proteins of TNF-α and NF-κB P50 on the femurs of such injected mice were lower than those of the control group (0.540±0.024 vs 0.682±0.031, 0.857±0.023 vs 1.098±0.044, n=3, P<0.05), while the expressions of BMP-2 were higher than the control group (0.99±0.04 vs 0.85±0.04, n=3, P<0.05). There was no difference in the PSmad1/5/8 expression on the bone of the two group of lupus mice (0.88 ±0.08 vs 0.84 ±0.04, n=3, P>0.05). The ectopic bone formation of BMMSCs of the etanercept injected MRL/lpr mice was higher than that of the normal saline injected mice, however, it was lower than that of the C3He/HeJ mice. Conclusion TNF-α inhibits osteoblast differentiation of mesenchymal stem cells by depressing Smad signaling which may contribute to the osteoporosis of the lupus mice.

Objective:To explore the effect of Naoshuning on the protein expression of Matrix metalloproteinases(MMPs) in experimental injuried brain tissue of rats. Methods:Immunohistochemistry was used to determine the changes of protein expression of MMPs. Brain tissue water content,permeability and ultramicrostructure of blood-brain barrier(BBB) were also observed. Results:Compared with the sham group,the brain tissue water and EB content of injured side and the level of MMP-2 and MMP-9 protein expression in brain tissue around contusion in model group increased obviously(all P

OBJECTIVE To investigate theradiosensitization of combined inhibition of signal transducer and activator of transcription 3(STAT3) and hypoxia-inducible factor-1α(HIF-1α)on laryngeal squamous carcinoma of xenograft mice and its underlying mechanisms.METHODS Xenograft mice were divided into 5 groups randomly: control group(A), irradiation group(B), irradiation and AG490 group(C), irradiation and PX478 group(D), irradiation combined AG490 and PX478 group(E). The size of xenograft tumor was measured and calculated. The expression of Ki67 and HIF-1α was detected by immunohistochemical method. Western blot was used to detect the expression of PARP1.RESULTS The size of xenograft tumor in group E was smaller compared with that in group C and group D. There were significantly difference between them respectively (t=12.367,11.598,P=0.000). The expression of HIF-1α in group E was lower than that in group C and group D respectively, and there were significantly difference respectively(t=5.422, 3.000,P<0.05). Ki67 index in group E was lower compared with that in group C and group D respectively and there were significantly difference respectively (t=4.479, 4.352,P<0.05). The level of cleaved PARP-1 in group E was higher than that in group C and group D respectively and there were significantly difference respectively (t=5.507, 7.102,P<0.05). CONCLUSION Combined inhibition of STAT3 and HIF-1α can increase the radiosensitivity of human laryngeal squamous carcinoma in the xenograft mice.