Objective To explore the clinical value of transvaginal color Doppler ultrasound(TV-CDS)in the detection of blood flow changes within the ovarian stromal artery in patients with ovarian endometriosis.Methods Blood flow indices within the ovarian stromal artery were measured by TV-CDS in 60 patients and60 normal controls.Results In ovarian endometriosis group,TV-CDS examination showed the color signal pattern was dot-like with high-resistance ovarian stromal arterial flow which manifested significant higher resistance index(RI),pulsatility index(PI)and systolic/diastolic(S/D)ratio than those in normal group(P<0.01).Analysis on clinicopathologic data showed that cystic history and diameter were risk factors affecting the absence of ovarian stromal blood signal,while cystic history,diameter and category were associated with the significant difference of blood flow display area(P<0.05).Conclusions TV-CDS can be used as a non-invasive,convenient and sensitive method for assessing blood flow changes within the ovarian stromal artery,indicating ovarian interstitial damage as well as pathological conditions of ovarian endometriosis that contributes to clinical diagnosis and treatment.

Objective The aim of this study was designed to investigate the changes of macrophages and activated microglias in white matter damage (WMD) in premature infants and effects of allopurinol. Methods An animal model for WMD was established by bilateral carotid artery occulation (BCAO). Forty-two newborn SD rats (1 day old) were divided randomly into 3 groups: sham surgery group (Sham), BCAO group (BCAO) and allopurinol treated group (ALLO). Pathological changes were studied 7 days and 14 days after BCAO, respectively. Macrophages and activated microglias were detected by immunohistochemistry 7 days and 14 days after BCAO, respectively. Results In BCAO group, Ten cases had mild or severe rarefaction in the corpus callosum area, especially at the cingulum. Pathological changes of white matter were found in 4 cases in internal capsule. Eight cases had subcortex white matter rarefaction. The extent of white matter rarefaction in ALLO group was reduced significantly. Enlargement of bilateral ventricles was found in 6 of 8 cases in BCAO group. Compared to BCAD group [(3.27±0.73)%] the average ventricle size was reduced significantly in ALLO group [(2.44±0.71)%] (P<0.05). ED1 positive cells were found in corpus callosum,hippocampus, and internal capsule in all groups. BCAO group had more ED1 positive cells than the other two groups, and the staining extent in BCAO group was stronger than that in the other two groups. Conclusions BCAO could be used in newborn rats (1 day old) to establish a premature WMD animal model. Macrophages and microglias may play an important role in premature WMD. ALLO may have a potential protective effect on premature SD rat with ischemic WMD.

Objective To determine the protein and mRNA levels of interleukin-17 (IL-17) in the peripheral blood of patients with systemic lupus erythematosus (SLE), and to analyze the relationship between IL-17 and disease activity. Methods Twenty-six hospitalized SLE patients and twenty-one normal controls were studied. Plasma protein and mRNA of IL-17 in peripheral blood were measured by ELISA and Real-time RT-PCR respectively. Results IL-17 level and its mRNA level increased in SLE patients compared with that in normal controls. Plasma concentration of IL-17 showed a significant positive correlation with SLEDAI score and anti-dsDNA antibody level, and a significant negative correlation with serum C3 level. Conclusions Plasma IL-17 protein and mRNA expression level in SLE patients increased significantly and had close relationship with disease activity, which might suggest that IL-17 play an important role in the pathogenesis of SLE.

Objective To determine the optimized self micro-emulsifying drug delivery system (SMEDDS) formulation of probucol. Methods According to the indexes of mean particle size, zeta-potential, solubility of probucol in blank SMEDDS and the dissolution percentage in 5 minutes of the preparations, the optimized formulation was determined by the central composite design-response surface methodology. Results When the correspondent percentage of olive oil in oil phase (W/W) was 0.33, the percentage of oil phase in formulation (W/W) was 0.5, and the ratio of surfactant to co-surfactant (W/W) was 2.0, respectively. The mean particle size, zeta-potential, solubility of probucol and dissolution percentage in 5 minutes of micro-emulsion was 92.7 nm, -17.38 mV, 65.17 mg/mL and 63.46%, respectively. Conclusions The optimized formulation of the probucol SMEDDS was obtained quickly and conveniently by the central composite design-response surface methodology. The method had a reliable predictability.

Objective To investigate the changes of gene expression in the cardiac outflow tract (OFT) in Cx43 knockout (Cx43-/-) mouse embryos, and to elucidate the genes involved in the myocardialization of proximal cardiae outflow tract septum. Methods The cDNA was retrotranscripted from RNA, which extracted from OFT tissues of both Cx43-/- and Cx43 wild type (Cx43+/+) mouse embryos on embryonic day (ED) 14.5. The biotin-labeled cRNA derived from the transcription of cDNA was fragmented as probes. The probes were hybridized with Affymetrix Mouse Genome 430 2.0 Array. Gene Array Scanner was used to screen the signals of hybridization, and the expression of genes was detected. Results Among the differentially expressed genes, there were 143 upregulated and 235 downregulated in Cx43 knockout OFT tissue compared with those in Cx43+/+ heart. Functions of proteins encoded by the altered genes encompassed all functional categories, with the largest percentage in genes involved in signaling pathways such as regulation of transcription, cell cycle, etc. Among the differentially expressed genes in the Cx43-/- heart, some were related with TGFβ/BMP signaling pathway, and Ssr1, Ptk2 and Bmp6 were related with conotruncal defects. These genes were verified by Real-time PCR, and the result was consistent with that of microarray (P<0.05).Conclusions Scaned by Gene Array and verified by Real-time PCR, genes related with TGFβ/BMP signaling pathway, and Ssr1, Ptk2 and Bmp6 were differentially expressed in ED 14.5 Cx43-/- OFT tissue, which may be involved in the myocardialization of proximal cardiac outflow tract septum.

Objective To establish a zebrafish IGFBP-2 gene knock-down model by morphilino modified antisense oligonucleotide injection, so as to investigate the abnormal phenotypes of heart and vessels in early stage of zebrafish development and the expression of zebrafish cardiogenesis related genes. Methods The spatiotemporal expression of IGFBP-2 gene in early stage of zebrafish development was testified by whole mount in situ hybridization with antisense RNA IGFBP-2 probe. The IGFBP-2 morpholino (IGFBP-2 MO) that especially inhibited the gene promoter and standard control morpholino (Con-MO) were designed and synthesized by Gene-tools Corporation. Four different concentration gradients (0.05, 0.10, 0.25 and 1.0 mmol/L) were set as IGFBP-MO injection groups with 0.25 mmol/L Con-MO injection group and wild type group as controls. Contribution to the incidence of heart abnormal phenotypes and mortality rate induced by 4 different IGFBP-2 concentrations injection group was recorded and compared with 2 control groups. Heart abnormal phenotypes at different developmental stages in 0.25 mmol/L IGFBP-2 injection group were observed in detail. To validate the effectiveness of IGFBP-2 MO, the expression of enhanced green fluorescence presented by wild type zebrafish embryos at 12hpf which received single injection of IGFBP-2 EGFP recombinant plasmid and those co-injected with Con-MO or IGFBP-2 MO were detected. To investigate the regulation relationship between IGFBP-2 gene and other cardiogenesis related genes, expression of atrium specific marker gene Amhc was detected in IGFBP-2 MO and wild type group by in situ hybridization. Ventricle specific green fluorescence of Vmhc-EGFP transgenic zebrafish embryos whose IGFBP-2 gene was knocked-down were compared with those untreated. Zebrafish peripheral vascular development in the IGFBP-2 MO group was also checked out by micro-angiography. Results Whole mount in situ hybridization revealed that IGFBP-2 gene expressed in turn at eyes, midbrain and then focused on liver in early stage of zebrafish development. The micro-injection of 0.25 mmol/L IGFBP-2 MO resulted in heart malformation in nearly 60% of all injected zebrafish embryos. Heart malformation phenotypes included slow heart beat, pericardial edema, weak ventricle systole contraction and heart tube looping disorder. Some of them represented atria dilation, blood regurgitation and ciculation obstruction. Wild type zebrafish embryos that received single injection of IGFBP-2 EGFP plasmid DNA or co-injected with Con-MO presented strong enhanced green fluorescence at 12hpf, meanwhile, the fluorescence was barely seen in the embryos co-injected with IGFBP-2 MO. This strongly validated the gene specific knock-down effect of IGFBP-2 MO. Amhc was down-regulated at 48hpf in IGFBP-2 MO group. Vmhc-EGFP transgenic zebrafish down-regulated by IGFBP-2 gene also resulted in attenuated expression of ventriclar-specific green fluorescence protein at 48hpf. Intersegmental blood vessels of IGFBP-2 MO group by micro-angiography at 60hpf demonstrated an sparsate and chaos image, which suggested that IGFBP-2 gene expression was involved in the regulation of normal vascular development. Conclusions Micro-injection of IGFBP-2 MO is an efficient way to knock-down IGFBP-2 gene in zebrafish embryos. IGFBP-2 gene expression down-regulation leads to heart and vessels maldevelopment and have an impact on the expression of cardiogenesis related genes of zebrafish embryos as well. In short, IGFBP-2 plays a critical role in the normal cardiovascular development of zebrafish embryos.

Objective To investigate the role of p38MAPK in the differentiation of murine osteoblasts, and to observe the expressions of receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG). Methods The calvarial osteoblasts of newborn BALB/c mice were cultured in MEM medium containing 10% NCS. Raloxifene (10~(-7) mol/L) and 17β-estradiol (10~(-8) mol/L) were added respectively when cells reached 70%-80% confluence combined with or without 5 μmol/L SB202190, an inhibitor of p38MAPK. The osteoblasts alkaline phosphatase activity assays were performed 72 hours later using PNPP method, and mRNA levels of alkaliphosphatase (ALP), OPG and RANKL were determined by RT-PCR. Results 17β-estradiol and raloxifene increased ALP activity and ALP mRNA level in osteoblasts in vitro which were blocked by p38MAPK inhibitor.The mRNA levels of RANKL and OPG were up-regulated by 17β-estradiol and raloxifene while the ratio of OPG/RANKL kept constant. SB202190 (5 μmol/L) inhibited the highly expressed RANKL and OPG in osteoblasts, and obviously decreased the ratio of OPG/RANKL. Conclusions p38MAPK inhibition blocked the differentiation of osteoblasts and decreased the up-regulated OPG and RANKL expressions in osteoblasts significantly (P<0.05).

Objective To detect steroidogenic acute regulatory protein (StAR) expression in apolipoprotein E-deficient mice at different ages and serum lipid levels. Methods Nighty-six C57BL/6J and apoE-/- mice were enrolled, which were divided into 16 groups with 6 mice per group according to age (1 day, 1, 3, 5 months), sex and genotype (C57BL/6J and apoE-/-). The serum lipid levels in C57BL/6J and apoE-/- mice were detected by commercial kits. StAR mRNA and protein expressions in liver were detected by Real-time PCR and Western blot respectively. Results ApoE-/- mice had higher LDL-cholesterol and lower HDL-cholesterol compared with C57BL/6J mice of the same age and sex. StAR mRNA and protein expressions were decreased following with aging in C57BL/6J mice. However, in apoE-/- mice with higher lipid levels, StAR mRNA and protein expressions were changed with the lipid levels other than ages. StAR mRNA and protein increased in the early stage, and then decreased with the increasement of lipids levels. Conclusions StAR could affect lipids levels and may be an effective regulator for atherosclerosis and other cardiovascular diseases.

Tumor necrosis factor alpha (TNF-α) is a homotrimeric protein encoded within the major histocompatibility complex (MHC). TNF-α can bind its receptors in the body to play a role in immune regulation and to participate in the various pathophysiology processes including fever, inflammation, infection, wound healing and tumors necrosis. TNF-α, an important component of inflammatory pathways, is up-regulated in the synovial tissue, synovial fluid and serum of rheumatoid arthritis (RA) patients. In RA patients, TNF-α along with many kinds of tissue factors and matrix proteins together promotes the inflammatory response, abnormal apoptosis of synovial cells, pannus formation and cartilage and bone destruction, which maintain the sustainability of the progress of RA. Thus, TNF-α may be a clinical indicator of RA activity as well as an effective target for RA.

Liquichip is a new type of biochip emerging in the 1990s, which is integrated flow cytometry, laser technology, digital signal processing and traditional chemical technology with the greatest features of high-throughput and high-flexibility. Luminex has promoted new products from Luminex 100/200 to Flexmap 3D. This is an introduction to the development, principle, and application of Luminex liquichip.