BACKGROUND:Both in vivo and vitro microenvironment can influence the proliferation and differentiation of neural stem cells (NSCs). In addition, cell culture solution plays variable roles in cell proliferation and differentiation.OBJECTIVE:To develop a convenient and rapid method to promote NSC primary culture by modifying traditional serum-free medium. DESIGN, TIME AND SETTING:Randomized controlled cell trial was performed at Institute of Brain Science, Qingdao University Medical School from November 2005 to September 2006.MATERIALS:Ten Wistar rat of gestation for 12-16 days; cell suspension fron brain tissue of embryonic rat. METHODS:Cell suspension were seeded into four 50-mL culture flasks with cell density of 1×106 mL-1, and divided into 2 groups with 2 flasks in each group. The control cells (flasks A and B) were cultured in serum-free medium containing DMEM/F12, B27 (2%), basic fibroblast growth factor (20 μg/L), and epidermal growth factor (20μg/L), and the experimental cells (flasks C and D) were firstly cultured with DMEM/F12 containing fetal bovine serum (FBS, 5%), following by the same serum-free medium 2 to 3 days later. MAIN OUTCOME MEASURES:The proliferation of NSCs was observed by inverted microscopy and immunocytochemistry.RESULTS:①Most of the cells in two culture conditions expressed nestin, the specific marker of NSCs, and were immunocytochemically positive. ②Neural stem cells cultured with FBS formed neurospheres 3-4 days earlier than those without FBS. ③There were no significant differences in cell number, but the neurospheres under the experimental condition were larger than those in control group. Some of the cells were neurone specific enolase-positive after serum-conditioned induction, and some were glial fibrillary acidic protein-positive, indicating NSCs had differentiated into neuron-like cells and neurogliocytes.CONCLUSION:FBS-conditioned pre-culture can accelerate the proliferation of neural stem cells.

Objective To manufacture magnetic microbubbles with dual-response to ultrasound and magnetic fields.Methods Microbubbles of ultrasound contrast agent (ST68) based on a surfactant were prepared by the acoustic cavitation method.Fe3O4 magnetic nanoparticles with negative charge were synthesized using the polyol procedure.Magnetic microbubbles were generated by depositing polyethylenimine and Fe3O4 magnetic nanoparticles alternately onto the microbubbles using the layer-by-layer self-assembly.In vitro ultrasonography was performed on a silicone tube with/without magnetic microbubbles (3 × 108/ml) by a self-made device to observe the movement of magnetic microbubbles under the effects of magnetic field.In vivo imaging was performed on the kidney of New Zealand rabbits before and after the injection of magnetic microbubbles.Results The Fe3O4 nanoparticles carried a stable negative charge of (-24.6 ± 6.7) mV and more than 98％ of the particles were less than 8 μm in diameter,meeting the size requirement of an ultrasound contrast agent for intravenous administration.There was no echoic signal in the silicone tube before injection of magnetic microbubbles,but there were strong echoic signals after injection.After applying a magnetic field,the magnetic microbubbles moved along the direction of the magnetic flux.In vivo ultrasound imaging could not visualize the kidney before injection of magnetic microbubbles,but could remarkably visualize the kidney after injection.Conclusions The magnetic microbubbles exhibit favorable magnetic targeting and ultrasound contrast enhancement characteristics.Such properties may serve as the foundation to study their potential for simultaneous diagnosis and treatment in the future.

Objective To fabricate an ultrasound/fluorescence bi-modal contrast agent by encapsulating fluorescent quantum dots into polymeric ultrasound contrast agent microbubbles.Methods Polylactic acid (PLA,500 mg),(1R)-(+)-camphor (50 mg) and CdSe/ZnS quantum dots (0.5 ml,2.3 μmol/L)were dissolved or dispersed in dichloromethane (10 ml) to form in an organic phase.Ammonium carbonate solution and poly (vinyl alcohol) solution were employed as the internal and external water phase,respectively.The fluorescent microbubbles were generated using double emulsion solvent evaporation and lyophilization methods.The morphology and illumination were characterized by scanning electron microscopy (SEM) and fluorescence spectrophotometry.Synchronized contrast-enhanced ultrasound and fluorescence imaging was acquired by injecting fluorescent microbubbles into the silicone tube coupled to a self-made ultrasound/fluorescence imaging device.Ultrasound/fluorescence bi-modal in vivo imaging was acquired on the kidney of New Zealand rabbits and suckling mice.Results The fluorescent microbubbles were hollow spheres with an averaged diameter of (1.62 ± 1.47) μm.More than 99％ of these microbubbles were less than 8 μm in diameter,which meeted the size criteria for ultrasound contrast agents.The fluorescence emission peak of the microbubbles appeared at 632 nm,indicating that good luminescence properties of quantum dots were maintained.In vitro ultrasound/fluorescence imaging showed no echoic signal when the silicone tube was filled with saline,but there was a strong echo when filled with fluorescent microbubbles.The liquid column with fluorescent microbubbles emitted red luminescence under ultraviolet irradiation.The kidney of the rabbit was remarkably enhanced after the administration of fluorescent microbubbles.Bright fluorescence could be observed at the injection site of the suckling mice via subcutaneous injection.Conclusions A bi-modal but single contrast agent based on polymeric microbubbles has been successfully fabricated for the use of ultrasound and fluorescence imaging.It retains the good characteristics of both echogenicity and fluorescence,which complement each other in case of limitations imposed by uni-modal,single agents.

BACKGROUND: Thrombus formation and neointimal hyperplasia limit its use for revascularization of small-caliber vessels (<6mm diameter) in the coronary or peripheral circulation.OBJECTIVE: To improve hemocompatibility, the luminal surface of a small diameter expanded polytetrafluoroethylene (Eptfe) vascular prosthesis was modified with alginate and recombinant hirudin.DESIGN: Observational experiment.SETTING: This study was performed in Nanomedicine and Biosensor Laboratory, Bio-X Center, Harbin Institute of Technology from Marcy 2006 to June 2007.MATERIALS: The GORE-TEX Epife vascular grafts (W. L Gore & Associates, Inc., Flagstaff, AZ) used in this study were 4mm in internal diameter. rHir was obtained from Calbiochem, Germany. Sodium alginate (also called alginic acid sodium salt; medium viscosity) was purchased from Sigma.METHODS: A p-diazonium diphenyl amine polymer (PA) was used as an interlayer between alginate and recombinant hirudin (rHir). The diazonium moieties were capable of covalently coupling with electron-rich aromatic systems such as histidine and tyrosine residues of hirudin. No need for chemical pretreatment took all advantage by preserving the bulk properties with almost no effect on stability and elasticity of the Eptfe vascular graft. rHir amount on Epife surface Was determined by the Micro BCA (Bicinchoninic acid) Protein Assay kit.MAIN OUTCOME MEASURES: ① Determination rHir amount on Eptfe surface; ② Static water contact angles; ③ Attenuated total reflectance fourier transform infrared spectroscopy (ATR-FTIR) was employed to confirm the change taking place in the Eptfe graft surface modification process; ④ Characterization of the surface morphology and platelet adhesion by SEM; ⑤ APTT and PT; ⑥ Percent hemolysis.RESULTS: ① The amount of rHir adsorbed onto the Eptfe vascular was deduced to be 16.35 μg/cm2. ② Surface analysis ATR-FTTR revealed the presence of new functional groups on the modified graft surfaces. ③ The water contact angle of the modified graft surface decreased. ④ The longer APTT and PT value lower than 5% hemolysis level and dramatically decrease of platelet adhesion assay showed that rHir modified graft had great improved blood compatibility.CONCLUS10N: Cross-linked Alg/rHir onto Eptfe can improve luminal surface and hemocompatibility.

OBJECTIVE To understand disinfection quality in newborn Spa room,to give prevention messures against nosocomial infection.METHODS The air,UV intensity,the object surface,staff′s hands,and the using of disinfectant were detected.RESULTS The total qualified rate was 98.61% and that of the object surface was 97.22%,of staff′s hands was 98.61%,of UV intensity and indoor air was 100% and of the using disinfectant was 97.22%.CONCLUSIONS Strengthening the disinfection,finding the weak timely and taking the effective measures can prevent the occurrence of hospital infection.

Objective To analyze the distribution of clinically isolated pathogenic bacteria in Xi′an area during 2014 and their drug resistant characteristics in order to provide the data of pathogenic bacterial drug resistance for medical pharmaceutical adminis ‐tration departments and clinical rational use of antibacterial drugs .Methods The pathogenic bacteria of nosocomial infections were cultured and isolated by using the routine method .The bacterial species was identified by using the semi‐automatic or full‐automatic bacterial identification and analysis systems .The drug susceptibility test was conducted according to CLSI standards .The data sta‐tistics and analysis were performed by using the WHONET 5 .6 software .Results 31 013 strains of pathogenic bacteria were isola‐ted in 2014 ,including 20 029 strains (64 .58% ) of Gram‐negative bacilli ,9 888 strains (31 .88% ) of Gram‐positive cocci and 1 096 strains (3 .54% ) of fungi ;the top bacteria was E .coli(20 .29% ) ,vancomycin resistant Staphylococcus aureus was not be found ;the resistance rates of Enterococcus faecium and faecalis against Vancomycin were 3 .00% ,1 .00% ,which against to linezolid was 1 .00% ;the generation rates of extended‐spectrum beta‐lactamase(ESBLs) in E .coli and Klebsiella pneumoniae were 65 .0% and 56 .0% respectively .Conclusion The important pathogenic bacteria ,including MRSA ,vancomycin resistant enterococcus ,carbapen‐em resistant Enterobacteriaceae bacteria ,pan‐drug resistant Pseudomonas aeruginosa and Acinetobacter baumannii ,in nosocomial infection should be performed the intensive monitoring and the communication with clinic should be strengthened in order to make the detection results serve the clinic well .

The modified early warning score (MEWS), as a rapid assessment and early warning scoring tool, has been widely used in patients in China, which can help nurses to identify potentially critical patients early, but not in all clinical fields. Through the retrieval of the relevant literature of the clinical application of the MEWS in Wanfang Medical Database from 2011 to 2018, the SWOT analysis was used in this paper [strength (S), weakness (W), opportunities (O) and threats (T)] methods, to systematically analyze the advantages, problems existence, implementation opportunities and challenges in the clinical application of the MEWS in our country. The purpose of this study was to obtain the best cut-off value of MEWS in different diseases and to formulate standard early-warning intervention measures for MEWS, which may provide reference for clinical workers to carry out relevant research.

Objective:To analyze the relationship between the developmental eye movement (DEM) test results and the vocabulary size in Chinese children with developmental dyslexia.Methods:A cross-sectional study was conducted.A total of 1 243 fifth grade students from 10 primary schools were enrolled from September to December 2019 in Tianjin, among which there were 664 males and 579 females, with the average age of (10.68+ 0.53) years old.The Chinese vocabulary test and intelligence test were carried out.Eighty-five dyslexic children with subaverage vocabulary size were selected as the experimental group and 54 normal children were selected as the control group.The DEM test was conducted in the two groups, and the vertical time, the horizontal adjustment time, the ratio of horizontal to vertical time and the total number of errors were recorded and analyzed.The differences in positive rate of dyslexia, various DEM test indicators between different genders and different groups were analyzed.The correlations between vocabulary size and vertical time, horizontal adjustment time, the ratio of horizontal to vertical time and the total number of errors were analyzed.This study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of School of Optometry, Tianjin Vocational Institute (No.ysgxyll001). Written informed consent was obtained from the guardian of each subject.Results:The total positive rate of dyslexia was 6.83%(85/1 243), and the positive rate of 9.33%(62/664) in boys was higher than 3.97%(23/579) in girls, with a significant difference between them( χ2=13.974, P<0.001). There were no statistically significant differences in age, vocabulary size, vertical time, horizontal adjustment time, and the ratio of horizontal to vertical time between different genders in the control group (all at P>0.05). The vocabulary size of girls in the experimental group was larger than that of boys, showing a statistically significant difference ( t=-2.259, P=0.027). There was no significant difference in age, vertical time, horizontal adjustment time, and the ratio of horizontal to vertical time (all at P>0.05). The vertical time and horizontal adjustment time of the experimental group were longer than those of the control group, and the differences were statistically significant ( t=-4.848, -4.297; both at P<0.001). There was no statistically significant difference in the ratio of horizontal to vertical time between the two groups ( t=0.126, P=0.900). The total number of errors was 0(0, 1) in the experimental group, which was higher than the control group 0(0, 0), with a significant difference between them ( H=1.979, P=0.001). The vocabulary size of students in the two groups was negatively correlated with the vertical time, horizontal adjustment time and the total number of errors ( r=-0.397, P<0.001; r=-0.355, P<0.001; r s=-0.180, P=0.034), and was not obviously correlated with the ratio of horizontal to vertical time ( r=0.038, P=0.656). Conclusions:The DEM test scores of Chinese children with developmental dyslexia are higher than those of normal children, and there is no difference between different genders.The lower the scores of vocabulary size test, the higher the scores of DEM test.

Objective:To assess plasma microfibrillar associated protein 5(MFAP5) level in patients with polycystic ovary syndrome(PCOS), and to explore its relationship with glucose and lipid metabolism as well as sex hormones.Methods:Fifty PCOS patients and 65 healthy female subjects were selected as PCOS group and control group, respectively. Clinical data and plasma MFAP5 levels between the two groups were compared.Results:The plasma MFAP5 level in PCOS group was significantly higher than that in control group( P<0.01), and the plasma MFAP5 level in PCOS overweight subgroup was higher than that in control subgroup( P<0.01). No difference was observed in plasma MFAP5 level between the two non-overweight subgroups( P>0.05). Correlation analysis showed that plasma MFAP5 level was positively correlated with waist circumference, low density lipoprotein-cholesterol, fasting insulin, homoeostasis model assessment of insulin resistance index(HOMA-IR), HbA 1C, testosterone, LH/FSH ratio, and leukocyte( P<0.05 or P<0.01). There was no significant correlation of MFAP5 with body weight, body mass index(BMI), hip circumference, waist hip ratio, high density lipoprotein-cholesterol(HDL-C), triglyceride, total cholesterol, and blood glucose( P>0.05). In PCOS group, plasma MFAP5 level was positively correlated with body weight, BMI, waist circumference, hip circumference, total cholesterol, and leukocyte( P<0.05 or P<0.01). There was no significant correlation of MFAP5 with waist hip ratio, HDL-C, triglyceride, blood glucose, fasting insulin, HOMA-IR, leukocyte, and sex hormones( P>0.05). Multivariate logistic regression analysis showed that MFAP5 was an independent risk factor for PCOS( P<0.05). Conclusion:Plasma MFAP5 level is increased in PCOS patients and is closely related to BMI, waist circumference, hip circumference, and total cholesterol. Plasma MFAP5 is an independent risk factor for PCOS, which may be involved in the pathogenesis of PCOS.

OBJECTIVE: To explore the genetic basis for a Chinese pedigree affected with pachyonychia congenita (PC). METHODS: With informed consent obtained, peripheral blood samples were taken from the pedigree. Genomic DNA was extracted with a phenol/chloroform method. Based on the clinical manifestation of the patients, candidate genes for PC were selected. Potential mutation was screened by PCR and Sanger sequencing. Suspected mutation was verified in other family members by PCR-high resolution melting (HRM) analysis. Haplotype analysis using microsatellite markers was also carried out to determine the founder of the mutation. RESULTS: A heterozygous c.275A>G (Asn92Ser) mutation was discovered in exon 1 of the KRT17 gene in the proband. PCR-HRM analysis showed that all affected members were heterozygous carriers of the mutation. The same mutation was found in none of the unaffected members. Haplotype analysis and sequencing indicated the mother of the proband to be the founder. CONCLUSION The c.275A>G (Asn92Ser) mutation of the KRT17 gene probably underlies the disease in this pedigree. Above finding has facilitated genetic counseling and prenatal diagnosis for this pedigree.
Asian Continental Ancestry Group ; Humans ; Keratin-17 ; genetics ; Mutation ; Pachyonychia Congenita ; genetics ; Pedigree ; Polymerase Chain Reaction