Adult ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Infertility, Female ; drug therapy ; Ovulation ; drug effects ; Ovulation Induction ; Phytotherapy

Objective To study the ability of silica nanoparticles as gene carriers for adult human epidermal keratinocytes gene transfection.Methods The silica nanoparticles-DNA conjugated with the enhanced green fluorescence protein plasmid DNA(pEGFP-N_1) was transfected into adult human epidermal keratinocytes.The expression of green fluorescence protein was investigated in transfected keratinocytes by eletromicroscope examine and the efficiency of gene transfection was revealed.Results The silica nanoparticles-DNA complexes can be effectively transfected into adult human epidermal keratinocytes and the efficiency of gene transfection was about 20%～30%.Conclusion The silica nanoparticles can be used as DNA carriers for gene transfection,and can efficiency transfect the pEGFP-N_1 into adult human epidermal keratinocytes.

Objective To explore the changes of fibrinolytic status and coagulation function of peripheral blood at the acute stage in patients with intracerebral hemorrhage(ICH).Methods The platelet(PLT) count and mean platelet volume (MPV),the levels of plasma fibrinogen(Fib) and D-dimer(D-D) were detected at

Objective To evaluate the effect of this neuroendocrine hormone on protein expression by treating the human dermal fibroblasts with a-melanocyte stimulating hormone (α-MSH ).Methods Thehuman dermal fibroblasts was cultured, and the total protein of the fibroblasts were separated with immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). After Coomassie bright blue staining, gel images were acquired by Image-scanner and then analyzed with the PDQuest software. 2-DE maps of fibroblasts were established. Partial differently expressed protein spots were incised from gels and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and MSDB database searching by Mascot? software were used for protein identification. Results Well-resolved, reproducible 2-DE patterns of dermal fibroblasts treated with and without crMSH were obtained. 8 differently expressed protein spots were detected, among which 8 obtained peptide mass fingerprints (PMF) by MALDI-TOF-MS analysis. Among these proteins, of particular interest were five proteins annexin I, HSP27 and lamin A, etc. Conclusions Proteins expressed by human dermal fibroblasts treated with or without crMSH are different, and some of the differently expressed proteins involve apoptosis, intracellular signal transduction and framework construction and so on, which may be associated with anti-fibrosis effects of (a)-MSH on human dermal fibroblasts.

Objective T o explore the effectiveness of direct am plification for the ST R analysis of carti-lage, and to accelerate the effectiveness of disaster victim identification. Methods E ighty-eight cartilage sam ples w ere directly am plified by Pow erPlex誖21 kit, and the results of genotyping w ere com pared w ith that obtained by the m agnetic beads m ethod. Results In 88 cartilage sam ples, the ST R genotypes w ere successfully detected from 84 sam ples by direct am plification and m agnetic beads m ethod, and both the results of genotyping by tw o m ethod w ere consistent. Conclusion D irect am plification w ith Pow er-Plex誖21 kit can be used for ST R genotyping of cartilages. T his m ethod is operated easily and prom ptly, w hich has a potential application in the individual identification of m ass disasters.

Objective To evaluate the therapeutic effects of aminoguanidine(AG) on cerebral ischemia-reperfusion damage in rats. Methods The intravascular thread models with 2 h of occlusion and 22 h of reperfusion were made in the rats.The brain infarction size and the degree of blood brain barrier(BBB) disruption in the ischemic regions were evaluated by staining with 2,3,5-triphenyl tetrazolium chloride and observing with Evans blue fluorescence microscope.HE staining was utilized for observing neutrophil infiltration. Results The brain infarction(volume,) the area of BBB disruption and the degree of neutrophil infiltration were dramatically decreased in the treatment group as compared to the control group(P

Alzheimer' s disease (AD) is the most common form of dementia in the elderly. AD is an invariably fatal neurodegenerative disorder with no effective treatment. Senescence-accelerated mouse prone 8 (SAMP8) is a model for studying age-related cognitive impairments and also is a good model to study brain aging and one of mouse model of AD. The technique of cDNA microarray can monitor the expression levels of thousands of genes simultaneously and can be used to study AD with the character of multi-mechanism, multi-targets and multi-pathway. In order to disclose the mechanism of AD and find the drug targets of AD, cDNA microarray containing 3136 cDNAs amplified from the suppression subtracted cDNA library of hippocampus of SAMP8 and SAMR1 was prepared with 16 blocks and 14 x 14 pins, the housekeeping gene beta-actin and G3PDH as inner conference. The background of this microarray was low and unanimous, and dots divided evenly. The conditions of hybridization and washing were optimized during the hybridization of probe and target molecule. After the data of hybridization analysis, the differential expressed cDNAs were sequenced and analyzed by the bioinformatics, and some of genes were quantified by the real time RT-PCR and the reliability of this cDNA microarray were validated. This cDNA microarray may be the good means to select the differential expressed genes and disclose the molecular mechanism of SAMP8's brain aging and AD.
Aging ; genetics ; metabolism ; Alzheimer Disease ; genetics ; metabolism ; Animals ; Disease Models, Animal ; Gene Expression ; Gene Expression Profiling ; Hippocampus ; metabolism ; Male ; Mice ; Models, Animal ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction

Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.
Animals ; Base Sequence ; Cloning, Organism ; Electrophoresis ; Endonucleases ; genetics ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; Gene Targeting ; methods ; Goats ; Green Fluorescent Proteins ; genetics ; Molecular Sequence Data ; Mutation ; Zinc Fingers

Objective To evaluate the NLRP7 gene mutations and variants and their expression of genetic approach in hydatidiform mole patients with family history.Methods Six cases of mole patients with family members of mole history and 60 healthy women, taking blood, extracting DNA, the genetic mutation on NLRP7 screening and analysis, looking for mutations and corresponding amino acids, proteins control gene mutation found NLRP7 area.Results In 6 mole patients with family history:three patients were with sister's history of mole, and 2 of them familial recurrent hydatidiform mole(from family MoCh76 and family Ch77), there are 2 loci NLRP7 gene mutation.Screening patients from family MoCh76 for mutations in NLRP7 revealed in exon 3 and exon 5, amino acids [295G ＞ T] and [1970A ＞ T], proteins [Glu99X] and [Asp657Val], in a heterozygous.Screening patients from family Ch77 for mutations in NLRP7 revealed in exon4 and exon 7, amino acids [1294C ＞ T] and [2471 + 1G ＞ A], proteins [Arg432X] and [Leu825X], in a heterozygous.Screening patients from family 105 for mutations in NLRP7 revealed no NLRP7 gene mutation.There were mother's history of mole in three patients, and they were not familial recurrent hydatidiform mole.Screening patients from family MoCh73 for mutations in NLRP7revealed in exon 4, amino acids [1137G ＞ C], proteins [Lys379Asn], in a heterozygous.Screening patients from family 106 and family 110 for mutations in NLRP7 revealed no NLRP7 gene mutation.There were not found mutations and variations in 60 cases of ethnic matched control group.Conclusion NLRP7 mutations may be lead to familial recurrent hydatidiform mole.

Objective To investigate the significance of change of fluid-attenuated inversion recovery(FLAIR) hyperintense vessel sign(HVS) after endovascular recanalization in acute ischemic stroke. Methods The clinical and imaging data of the patients with acute middle cerebral artery(MCA) occlusion treated by mechanical thrombectomy with Solitaire AB from January 2013 to october 2014 were analyzed retrospectively. The inclusion criteria: (1) The preoperative MRI included conventional non-enhanced MR, diffusion-weighted imaging (DWI), magnetic resonance angiography(MRA) and perfusion-weighted imaging (PWI), and HVS was observed on preoperative FLAIR images; (2) acute MCA occlusion verified by conventional angiography;(3) postoperative similar MR images examination was performed within 48 hours. The relationships among postoperative changes in the HVS, DWI and Thrombolysis In Cerebral Ischemia (TICI) scale (1—3) were assessed. Results After endovascular therapy, HVS of the 11 cases were showed to be disappeared(n = 9) and decreased (n = 2). All the 9 patients with disappeared HVS achieved high grade flow (TICI 3), and minor decrease of ischemic area on DWI in 1 case, minor progression in 6, and significant progression in 2. However, of the 2 patients with decreased HVS, one achieved relatively low grade flow (TICI 2a) and the other was found to be relatively high grade flow (TICI 2b), but severe MCA stenosis. DWI demonstrated significant progression in both two cases. Conclusion Our data indicate that endovascular recanalization of acute MCA occlusion was effective for decreasing HVS. Postoperative decrease and disappear in HVS can be considered as a marker for hemodynamic improvement.