Objective To evaluate the feasibility of obtaining good quality ultrasound pictures of sutures and anterior fontanelle with three-dimensional and two-dimensional ultrasound. Methods Eighty fetuses at 16-35 weeks of gestation were evaluated with two-dimensional and three-dimensional ultrasound. The sagittal, coronal, lambdoidal and metopic sutures, as well as anterior fontanelle, were inspected. Results The visualization of the sagittal suture of three-dimensional ultrasound was significantly superior to that of two-dimensional ultrasound, while no significant difference in visualization of the metopic, lambdoidal, coronal sutures and anterior fontanelle was found between the two modalities. The visualization of all sutures and anterior fontanelle was significantly superior with three-dimensional ultrasonography to that of two-dimensional ultrasonography before 30 weeks of gestation, whereas no significant difference was found between the two modalities after 30 weeks of gestation. Conclusion Three-dimensional ultrasound can provide more comprehensive information of fetal cranial sutures and anterior fontanelle than two-dimensional ultrasound does. Three-dimensional ultrasound can also supply high visualization of all sutures and anterior fontanelle before 30 weeks of gestation.

Objective To assess the application value of three-dimensional (3D) ultrasound in visualizing fetal cranial sutures and fontanels. Methods One hundred women at different pregnant ages (16-19~+ weeks, 20-24~+ weeks, 25-29~+ weeks, 30-34~+ weeks, respectively) underwent 3D ultrasound, in order to observe the cranial sutures and fontanels of fetuses. Results ①Most cranial sutures and fontanels (70.00%-98.00%) could be visualized with 3D ultrasound. However, the sagittal suture revealed a lower displaying rate (53.00%) than others. ②At different pregnant ages, there was no significant difference of displaying rate of the metopic, coronal, squamoal sutures and the sphenoid, mastoid fontanels between fetuses and there was significant difference of displaying rate of the anterior and post fontanels, sagittal and lambdoid sutures. Conclusion 3D ultrasound can be used as an effective method for visualizing fetal cranial sutures and fontanels.

Objective To construct the pGEX-4T-2-CblN/Grb2, express and purify the corresponding chimeric protein; To investigate whether the chimeric CblN/Grb2 possesses ubiquitin ligase activity. Methods Total RNA of SK-BR-3 cells were isolated and reversely transcribed into cDNA, which were used as templates to amplify Grb2 SH2 by PCR. The gene fragment encoding the N-terminal part of Cbl protein (named as CblN) was amplified by PCR using pEFHACbl plasmid encoding human Cbl as templates. BamH I and EcoR V restriction enzyme digestion sites were introduced into both flanks of SH2 by overlapping extension PCR and the modified gene were cloned into pcDNA3.1(+). The pcDNA3.1(+)-CblN/Grb2 was obtained by replacing SH2 of CblN with Grb2SH2 and then used as templates to amplify CblN/Grb2 by PCR. The pGEX-4T-2-CblN/Grb2 was constructed by subcloning CblN/Grb2 into the prokaryotic expressing vector pGEX-4T-2. The GST-CblN/Grb2 fusion protein was expressed in E. coli of DH5? under IPTG induction and further purified with Glutathione Sepharose 4B. In vitro ubiquitination assay was performed to investigate whether the GST-CblN/Grb2 fusion protein is able to mediate auto-ubiquitinating reaction, namely whether it possesses ubiquitin ligase activity. Results The fusion expressing vector of pGEX-4T-2-CblN/Grb2 was successfully constructed; The GST-CblN/Grb2 fusion protein was correctly expressed and purified; In vitro ubiquitination assay indicated that GST-CblN/Grb2 fusion protein is able to mediate auto-ubiquitinating reaction and therefore possesses ubiquitin ligase activity. Conclusion Expression, purification of GST-CblN/Grb2 and identification of its′ activity have laid the foundation for further study of chimeric ubiquitin ligase′ effects on the growth of HER2 positive tumor cells.

AlM: To investigate the effect of exogenous peroxisome-proliferator-activated receptor-γcoactivator-1α ( PGC-1α) on vascular endothelial growth factor ( VEGF) expression in human retinal vascular endothelial cells ( HRVEC) .METHODS:Recombinant PGC-1α protein was added to HRVEC, and no recombinant PGC-1α protein was added to HRVEC as control group. After 24h of incubation, two groups of cells were then placed into a normoxic ( 20%O2 ) or hypoxic ( 1% O2 ) environment for another 16h. The expression of VEGF mRNA and protein were detected by real - time PCR, ELlSA and immunofluorescence cytochemistry.RESULTS: VEGF mRNA and protein levels in the cells were significantly increased by recombinant PGC - 1αprotein both under normoxia and hypoxia conditions as compared with control groups (P<0. 01).CONCLUSlON: PGC-1α can upregulate the expression of VEGF in HRVEC under normoxia and hypoxia conditions.

Objective To evaluate the inhibitory effect of small interfering RNA (siRNA) targeting peroxisome-proliferator-activated receptor-γ coactivator-1α (PGC-1α) on retinal neovascularization in the mouse.Methods Eighty seven-day-old C57BL/6J mice were divided into normal group,model blank group,model control group and PGC-1α siRNA group,twenty mice in each group.Mice in the normal group were kept in normal room air.Mice in the model blank group,model control group and PGC-1α siRNA group were induced for retinal neovascularization by hypoxia.Liposome with PGC-1α siRNA (1 μl) and liposome with negative control siRNA (1 μl) were injected into the vitreous in the PGC-1α siRNA group and model control group respectively when mice were moved out to room air from the cabin (Postnatal 12).No injection were performed in the model blank group.At postnatal 17,fluorescein angiography was used to assess the vascular pattern.The proliferative neovascular response was quantified by counting the nuclei of new vessels extending from the retina into the vitreous in cross-sections.PGC-1α and vascular endothelial growth factor (VEGF) level in retina were measured by real-time polymerase chain reaction (real-time PCR) and Western blot.Inhibition efficiency of PGC-1α siRNA on PGC-1α and VEGF was calculated.Results Mice in the normal group showed reticular distribution of retinal blood vessels.Central nonperfused retina,neovascular tufts and fluorescein leakage were seen in the model blank group and model control group.Neovascular tuft and fluorescein leakage were decreased in the PGC-1α siRNA group compared to the model blank group and model control group.The neovascular nuclei were increased in the model blank group and model control group compared to the normal group (P＜0.05).The neovascular nuclei were decreased in the PGC-1α siRNA group compared to the model blank group and model control group (P＜0.05).The expression of PGC-1α mRNA and protein in retina was increased significantly in the model blank group and model control group as compared with normal group,while decreased 54％ and 53％ respectively in the PGC-1α siRNA group as compared with model blank group and model control group (P＜0.05).The expression of VEGF mRNA and protein in retina was increased significantly in the model blank group and model control group as compared with normal group,while decreased significantly in the PGC-1α siRNA group (decreased 48 ％ and 40 ％ respectively) as compared with model blank group and model control group (P＜0.05).Conclusions Intravitreal injection of PGC-1α siRNA mediated by liposome can inhibit retinal neovascularization in the mouse effectively.

OBJECTIVE To monitor the sterilization effect of hydrogen peroxide gas plasma sterilizer with process challenge derice(PCD).METHODS Hydrogen peroxide gas plasma sterilizer was used to sterilize different material and structure items in simulation field sterilization trial.The test pieces have been cultured for 7 days at 37 ℃.Make the test records in detail.RESULTS The hemostatic forceps,surface of lines and biological indicators,as well as 300 mm stainless steels tube and 2000 mm Teflon tube were sterilized successful.But 600 mm and 300 mm stainless steels of low temperature did not past the challenge tests.The results of test surface and test lumen were inconsistent.CONCLUSIONS PCD is need to be introduced in monitoring sterilization effect of hydrogen peroxide gas plasma sterilizer.

Objective To explore the effect of Jiawei Wendan decoction on whole regulating function of CaMK/MAPK signal pathway in hippocampus of depression model rats.Methods Isolated depression model rats as research object were living unpredictable chronic stress.Based on the examination of mRNA/protein expression which was the key factor of the signal pathway of CaMK/MAPK and the intersection CREB1 mRNA expression,the relationship between CaMK Ⅱ/RSK protein expression and the CREB1 mRNA expression were analyzed with relation analysis method.Results ① Bidirectional correlation analysis:the respective coefficient (r value) of CaMK Ⅱ protein expression and the CREB1mRNA expression in hippocampus of the model group,Chinese medicine treatment group and western medicine treatnent group were-0.502 (P ＜ 0.01),-0.643 (P ＜ 0.01),-0.408 (P＜ 0.05) ;the respective coefficient(r value) of RSK protein expression and the CREB1 mRNA expression in hippocampus of the model group,Chinese medicine treatment group and western medicine treatment group were 0.550 (P ＜ 0.01),0.687 (P ＜ 0.0 l),0.407 (P ＜ 0.01).②Regression analysis:the respective regression coefficient of CaMK Ⅱ protein (positive direction),RSK protein (negative direction) and CREB1 mRNA expression in the model group,Chinese nedicine treatment group and western medicine treatment group were R 2 =0.472,F =12.983(P＜0.01),R2=0.666,F=28.961(P＜0.01),R2=0.356,F=8.004(P＜0.01).Conclusion The anti-depressant effect mechanism of Jiawei Wendan decoction is to regulate the whole function of the CaMK and MAPK signal pathway and then further up-regulate CREB1 mRNA expression.

BACKGROUND:Previous study has observed that the calf acellular dermal membrane exhibits slow repair efficiency, fast degradability speed and other shortcomings in the repair of cartilage defects. OBJECTIVE:To investigate the repair effect of the col agen type Ⅱ-modified acel ular dermal membrane on cartilage defects in rabbits. METHODS:The fetal rabbit chondrocytes were seeded onto the col agen type Ⅱ-modified acel ular dermal membrane, and the composite was then observed under scanning electron microscope at 3, 7 and 14 days. Cartilage defect models were established on the bilateral femoral condyles of 24 New Zealand white rabbits, and these model rabbits were randomly allocated to three groups. The cartilage-acellular dermal membrane and cartilage-collagen type Ⅱ-modified acellular dermal membrane were implanted into the defect regions of control and experimental groups, respectively. Those received no intervention were as blank control group. Collagen type Ⅱ immunohistochemical staining and Wakitani scoring system were performed at 6 and 12 weeks postoperatively. RESULTS AND CONCLUSION:Chondrocytes grew and adhered well in the scaffold. The Wakitani scores in the experimental group were significantly lower than those in the control and blank control groups at postoperative 6 and 12 weeks (P<0.05). At 6 and 12 weeks postoperatively, collagen type Ⅱ immunohistochemical staining was the strongest in the experimental group, with yellow and brown particles in the cytoplasm;the control group was positive for collagen type Ⅱ immunohistochemical staining, while the blank control group was negative for the staining. Our findings suggest that the collagen type Ⅱ-modified acellular dermal membrane is beneficial for the repair of cartilage defects.

Aged ; Humans ; Immunoglobulin G ; Lymphatic Diseases ; Male

Antineoplastic Agents ; administration & dosage ; therapeutic use ; Cladribine ; administration & dosage ; therapeutic use ; Humans ; Leukemia, Hairy Cell ; drug therapy ; Male ; Middle Aged