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.

Extremities ; Humans ; Joint Prosthesis

AIMTo study the chemical constituents of Polygala aureocauda Dunn..

METHODSChemical compounds were isolated by column chromatography and their structures were determined mainly by spectroscopic means (UV, IR, MS, 1HNMR, 13CNMR, HMQC, HMBC).

RESULTSThree compounds were isolated and identified as 3-hydroxy-1,4-dimethoxyxanthone (I), 1, 7-dihydroxy-2, 3-methylendioxyxanthone (II), 7-hydroxy-1-methoxy-2, 3-methylendioxyxanthone (III).

CONCLUSIONCompounds I-III were isolated from Polygala aureocauda Dunn. for the first time, whereas compound I is a new xanthone.

Molecular Conformation ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polygala ; chemistry ; Xanthones ; chemistry ; isolation & purification

Anesthesia ; Fiber Optic Technology ; Humans ; Intubation, Intratracheal

AIMTo study the active constituents of Swertia davidi Franch..

METHODSChromatography was used to isolate and purify the chemical components, their structures were identified by spectral analysis.

RESULTSThree compounds were identified as 1,7-dihydroxy-3,8-dimethoxyxanthone (gentiacaulein) (V), 1,8-dihydroxy-3,7-dimethoxyxanthone (methylswertianin) (VI) and 1,8-dihydroxy-3,4,7-trimethoxyxanthone (VII).

CONCLUSIONCompound VII is a novel xanthone, named daviditin A, the others were isolated from Swertia davidi Franch. for the first time.

Molecular Structure ; Plants, Medicinal ; chemistry ; Swertia ; chemistry ; Xanthones ; chemistry ; isolation & purification

Objective To investigate new nurses' emotional state and its influencing factors,so as to provide references for their emotion management.Methods General information questionnaire,emotional state scale and nurse job stressors questionnaire were used to investigate emotional state and influencing factors of 120 new nurses from six Grade A hospitals in Beijing.Results Totals of 120 questionnaires were handed out and 103 were recovered,with the rate of 85.83％,among which 93 were valid,with the rate of 90.29％.The score of tension and anxiety,depression,anger and hostile,vigor and hyperactivity,fatigue and inertness,confusion and lost were respectively (17.23 ± 3.19),(33.75 ± 4.10),(27.67 ± 3.63),(17.24 ± 2.06),(14.38 ± 2.48),(13.16 ±2.20)in 93 new nurses,and (6.40 ±4.00),(7.20 ±8.20),(8.60 ±6.10),(19.80 ±4.60),(6.10 ± 3.40),(4.30 ± 2.70) in ordinary people,and the differences were statistically significant (t =32.69,62.44,50.60,-12.00,32.13,38.87,respectively; P ＜0.01).Issues concerning patients' nursing were the influencing factors for their tension and anxiety,depression,anger and hostile,as well as fatigue and inertness (t =4.94,4.33,3.29,3.77,respectively; P ＜0.01).Too heavy workload was the other reason for new nurses' fatigue and inertness (t =3.42,P ＜ 0.01).Management and interpersonal relation issues were the influencing factors for their anger and hostile,vigor and hyperactivity (t =3.89,-2.71,respectively; P ＜ 0.01).Nursing profession and work-related problems were the main influencing factors of new nurses' confusion and lost (t =5.19,P ＜0.01).Conclusions New nurses' emotional state is not good in a whole,and nursing managers should conduct specific emotion management according to different reasons.

Fibroblast growth factor 21 (FGF21) is a member of FGF family. It has been demonstrated that FGF21 is an independent, safe and effective regulator of blood glucose levels in vivo. In order to improve the activity of FGF21, we exchanged the beta10-beta12 domain of the human FGF21 with that of the mouse FGF21 to construct a novel FGF21 gene (named hmFGF21), and then subcloned hmFGF21 gene into the SUMO expression vector to create pSUMO-hmFGF21 and transformed it into E. coli Rosetta for expression of the fusion protein SUMO-hmFGF21. Both in vitro and in vivo glucose regulation activity of hmFGF21 was evaluated. The SDS-PAGE result showed that compared with wild-type hFGF21, the soluble expression of hmFGF21 increased about 2-fold. HmFGF21 was more potent in stimulation of glucose uptake in HepG2 cells in vitro. The results of anti-diabetic effect on db/db mice demonstrated that hmFGF21 had better efficacy on controlling the blood glucose of the db/db diabetic animals than wild-type hFGF21. These results suggest that the biological properties of FGF21 are significantly improved by optimization.
Amino Acid Sequence ; Animals ; Blood Glucose ; metabolism ; Cysteine Endopeptidases ; Diabetes Mellitus, Experimental ; blood ; Endopeptidases ; genetics ; Escherichia coli ; Fibroblast Growth Factors ; genetics ; metabolism ; pharmacology ; Genetic Vectors ; Glucose ; metabolism ; Hep G2 Cells ; metabolism ; Humans ; Hypoglycemic Agents ; metabolism ; pharmacology ; Male ; Mice ; Mutation ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Transformation, Genetic

Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.
Amino Acid Sequence ; Animal Diseases/*virology ; Animals ; Animals, Zoo/*virology ; Coronaviridae Infections/*veterinary/virology ; Coronavirus, Canine/genetics/*isolation & purification ; Fatal Outcome ; Female ; Male ; Molecular Sequence Data ; Sequence Alignment ; Sequence Homology, Amino Acid ; Ursidae/*virology ; Viral Proteins/chemistry

AIMTo study the active constituents of Swertia davidi Franch.

METHODSColumn chromatographies on silica gel, Sephadex LH-20 and Diaion-201 et al. were used to isolate and purify the chemical components. Their structures were identified by UV, IR, MS, NMR and 2D-NMR.

RESULTSThese compounds were elucidated as 8-O-beta-D-glucopyranosyl-1, 3, 5-trihydroxyxanthone (I), 5-O-beta-D-glucopyranosyl-1, 8-dihydroxy-3-methoxyxanthone (II), 5-O-beta-D-glucopyranosyl-1, 3, 8-trihydroxyxanthone (III) and swertamarin (IV).

CONCLUSIONCompound III is a new xanthone glucoside. The other compounds were isolated from this plant for the first time.

Glucosides ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Swertia ; chemistry ; Xanthones ; chemistry ; isolation & purification

OBJECTIVETo study the effect of highly active antiretroviral therapy (HAART) on plasma levels of MSP and MCP-1 in AIDS patients.

METHODSForty Chinese AIDS patients were treated with HAART for 3 months and 84 German AIDS patients with HAART for 3 to 6 years. The pre-treatment and post-treatment plasma levels of MSP and MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA), and their correlations with CD4+ cell counts and viral loads were analyzed.

RESULTThe mean levels of MCP-1 were significantly higher and MSP were significantly lower in HIV-infected patients compared with the HIV-negative controls (P <0.01). After HAART for three months, there were no significant changes in the levels of these cytokines. But after long-term HAART (for 3 to 6 y), the level of MCP-1 was increased and that of MSP decreased significantly (P<0.01). There was a negative correlation between MSP and MCP-1 levels, and the same for MSP level and CD4+ cell counts; while there was a positive correlation between MCP-1 levels and CD4+ cell counts.

CONCLUSIONThe changed plasma levels of MSP and MCP-1 are associated with HIV-1 infection and HAART may reverse the levels of these two cytokines.

Acquired Immunodeficiency Syndrome ; blood ; drug therapy ; Adult ; Anti-HIV Agents ; therapeutic use ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Chemokine CCL2 ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Macrophage-Activating Factors ; blood ; Male ; Middle Aged ; Time Factors ; Treatment Outcome