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1.
Article in Chinese | WPRIM | ID: wpr-1020035

ABSTRACT

Objective:Induced pluripotent stem cells (iPSCs) cell lines were established using peripheral blood mononuclear cells (PBMCs) from a patient suffering from neonatal respiratory distress syndrome (NRDS) who carried Adenosine triphosphate-binding cassette transporter A3 ( ABCA3) compound heterozygous mutations. Methods:Cell experimental research.Peripheral venous blood was collected and PBMCs were isolated and cultured in vitro. PBMCs were transfected with non-integrated Sendai vector carrying reprogramming factors.The chromosome karyotypes of the established iPSCs were analyzed.Immunofluorescence and flow cytometry were used to detect pluripotency markers of stem cells and verify their differentiation potential.Sanger sequencing was performed to analyze gene mutations.In addition, short tandem repeat (STR) analysis was performed, polymerase chain reaction(PCR) and agarose gel electrophoresis were used to detect virus residual. Results:Karyotype analysis of established iPSCs cell lines showed normal diploid 46, XY karyotype.Immunofluorescence showed positive staining of stem cell pluripotency markers OCT4, SSEA4, Nanog and Sox2.Flow cytometry was used to detected stem cell pluripotency markers and showed expression of TRA-1-60, SSEA-4 and OCT4.After differentiation into all three germ layers, immunofluorescence was performed to detect ectoderm (Pax-6), mesoderm (Brachyury) and endoderm alpha-fetoprotein markers, and the results showed positive staining, which confirmed that the iPSCs had the potential to differentiate.Sanger sequencing showed c. 3997_3998del and c. 3137C>T compound heterozygous mutations.STR analysis showed they originate from PBMCs, and no Sendai virus residual was detected by PCR and agarose gel electrophoresis.Conclusions:In this study, PBMCs from patient carrying ABCA3 compound heterozygous mutations was used to establish iPSCs cell lines.The research lays a foundation for the study of pathogenesis, therapeutic drug screening and cell therapy of NRDS caused by ABCA3 gene mutations.

2.
Article in Chinese | WPRIM | ID: wpr-1030652

ABSTRACT

Objective To investigate the risk factors of infection of Pseudomonas aeruginosa combined with Elizabethkingia anophelis in patients with cerebral hemorrhage and the antimicrobial treatment plan. Methods Clinical pharmacists participated in the treatment of pulmonary infection caused by Pseudomonas aeruginosa combined with Elizabethkingia anophelis in a patient with cerebral hemorrhage. The risk factors of Elizabethkingia anophelis infection and antimicrobial treatment plan were analyzed by referring to literature and combining the patient’s condition, medical history, drug use history and related examination results. Results Based on the infection site, the characteristics of mixed bacterial infection, and the metabolic/pharmacodynamic characteristics of antimicrobial agents, clinical pharmacists made drug recommendations for clinicians in the adjustment of anti-infection protocols, and patients’ systemic infections were effectively controlled. Conclusion Elizabethkingia anophelis is a conditional pathogen with low virulence and is not easy to infect healthy people. When the patient's immunity is low, it is easy to transform into pathogenic bacteria, which should be paid attention to.

3.
Article in Chinese | WPRIM | ID: wpr-929815

ABSTRACT

ATP-binding cassette transporter A3(ABCA3) gene mutation is one of the important causes of severe respiratory distress syndrome and interstitial lung disease in children and adults.Clinical phenotypes vary dramatically among patients with ABCA3 mutations.So far, the genotype-phenotype correlation is not entirely clear.The association between the possible factors that influence the phenotypes, such as the environment, infection and diseases remains to be studied.There is no specific treatment for the diseases caused by the mutations.Present models for studying ABCA3 mutations in vitro are still to be improved.This article focuses on reviewing the structure, genetics and the research progress of treatment of ABCA3 gene mutation related pediatric diseases, in order to provide experience and ideas for further researches and treatment of the diseases caused by ABCA3 gene mutation.

4.
Article in Chinese | WPRIM | ID: wpr-708114

ABSTRACT

Objective To evaluate the geometric and dosimetric accuracy of autosegmentation software for contouring the organ-at-risk ( OAR) of esophageal cancer, and discuss its clinical feasibility. Methods A total of 10 patients were enrolled, and single and multi-template were adopted respectively to auto-delineate corresponding OARs on target CT images based on image registration. The geometric consistency including volume difference (ΔV) , dice similarity ( DSC) and position difference (Δx, Δy,Δz) between the two autosegmentation method and manual were compared using Wilcoxon signed-rank test. And the correlation between DSC and OAR volume was analyzed. In addition, to evaluate the clinical feasibility of autosegmentation, the dose distributions of all OARs were compared using Friedman test. Results The average DSC of all OARs obtained by single and multi-template were 0.82 ± 0.17 and 0.92 ± 0.54, respectively, with statistically significant difference (Z= -2.803- -2.497, P<0.05). A positive correlation between DSC of the autosegmentation and OAR volume was found by spearman analysis, and the single-template was not good enough for the spinal cord with smaller volume. The positional deviations of multi-template group were less than 0.5 cm in three directions, which were better than single-template group. The main dosimetric indexes of single-template and multiple-template were similar to manual coutours. V20 of whole lung were 23.2%, 22.4% and 22.1%, Dmeanof whole lung were (11.3 ±4.0), (11.1 ±4.5) and (11.0 ±4.3) Gy, Dmaxof spinal cord were (40.3 ±4.8), (38.2 ±6.7) and (39.4 ± 5.3) Gy, respectively, and V30 of heart were 16.0%, 15.8% and 15.5%, respectively. There was no statistical difference between the three methods (P>0.05), and all of the dosimetric indexes were in line with the requirements of clinical dose limits. Conclusions The autosegmentation software can achieve satisfactory precision for the OARs of the esophageal cancer patients, and the multi-template method is better than the single-template, which is more suitable for clinical application.

5.
Article in Chinese | WPRIM | ID: wpr-658860

ABSTRACT

Objective To simulate the possible systematic delivery errors introduced by monitor units ( MU) and multi-leaf collimator ( MLC) in radiotherapy plans for nasopharyngeal carcinoma ( NPC) , and to analyze the dosimetric sensitivity of static intensity-modulated radiotherapy ( IMRT ) and volumetric modulated arc therapy ( VMAT) with these errors. Methods Five IMRT plans were replanned using VMAT modality with the same physical parameters, and then MU errors of 125%, 250%, and 5. 00% were introduced into IMRT and VMAT plans. Meanwhile, to simulate leaf position errors during delivery, MLC position errors (025 mm, 050 mm, 100 mm, 150 mm, and 200 mm) were introduced by modifying the original plan documents. The types of MLC errors were as follows:( 1) the MLC banks moved in the same direction;( 2) the MLC banks moved in opposing directions ( expand or contract the MLC gaps ) . The differences in dosimetric sensitivity introduced by MU and MLC errors between IMRT and VMAT plans for NPC were calculated by linear regression analysis. Results With the increase in MU errors, the doses to target and organs at risk ( OARs) of IMRT and VMAT plans increased in a linear way, and met R2=0992-1000( P<005) . For MLC errors, the average dosimetric sensitivity for target and OARs of IMRT and VMAT were-026%/mm and-065%/mm in case of offset errors, 487%/mm and 868%/mm in case of expansion errors, and -604%/mm and -988%/mm in case of indentation errors. In addition, the dosimetric sensitivity with the three types of MLC errors was greater for VMAT plan than for IMRT plan. ConclusionsMU and MLC errors have a significant effect on the dose distribution of IMRT, and particularly VMAT, for NPC. It is important to execute routine quality assurance of MLC to ensure accurate radiotherapy.

6.
Article in Chinese | WPRIM | ID: wpr-661779

ABSTRACT

Objective To simulate the possible systematic delivery errors introduced by monitor units ( MU) and multi-leaf collimator ( MLC) in radiotherapy plans for nasopharyngeal carcinoma ( NPC) , and to analyze the dosimetric sensitivity of static intensity-modulated radiotherapy ( IMRT ) and volumetric modulated arc therapy ( VMAT) with these errors. Methods Five IMRT plans were replanned using VMAT modality with the same physical parameters, and then MU errors of 125%, 250%, and 5. 00% were introduced into IMRT and VMAT plans. Meanwhile, to simulate leaf position errors during delivery, MLC position errors (025 mm, 050 mm, 100 mm, 150 mm, and 200 mm) were introduced by modifying the original plan documents. The types of MLC errors were as follows:( 1) the MLC banks moved in the same direction;( 2) the MLC banks moved in opposing directions ( expand or contract the MLC gaps ) . The differences in dosimetric sensitivity introduced by MU and MLC errors between IMRT and VMAT plans for NPC were calculated by linear regression analysis. Results With the increase in MU errors, the doses to target and organs at risk ( OARs) of IMRT and VMAT plans increased in a linear way, and met R2=0992-1000( P<005) . For MLC errors, the average dosimetric sensitivity for target and OARs of IMRT and VMAT were-026%/mm and-065%/mm in case of offset errors, 487%/mm and 868%/mm in case of expansion errors, and -604%/mm and -988%/mm in case of indentation errors. In addition, the dosimetric sensitivity with the three types of MLC errors was greater for VMAT plan than for IMRT plan. ConclusionsMU and MLC errors have a significant effect on the dose distribution of IMRT, and particularly VMAT, for NPC. It is important to execute routine quality assurance of MLC to ensure accurate radiotherapy.

7.
Article in Chinese | WPRIM | ID: wpr-501869

ABSTRACT

Objective To test and evaluate the geometric accuracy of delineation of organs at risk ( OARs) in head and neck cancer using an atlas?based autosegmentation ( ABAS) software. Methods The atlases for the ABAS software was generated using images from 40 patients with head and neck cancer undergoing intensity?modulated radiotherapy. The software was tested in 40 new patients. Automatic delineation of OARs was carried out on computed tomography images by single?( one to one ) and multi?template ( ten to one) approaches. In order to evaluate the feasibility of the automatic delineation in clinical application, differences in volume (ΔV%), position (Δx,Δy, andΔz), conformability (sensitivity ( Se ), specificity ( Sp ) , and dice similarity coefficient ( DSC) ) , and delineation time were assessed between the automatic and manual delineation. The comparison between the two automatic delineation approaches was made by paried t test. Results For all OARs, the multi?template automatic delineation achieved a significantly smaller mean ΔV% value and a significantly larger mean DSC value than the single?template automatic delineation (-0.02%± 0?29% vs. -0.16%± 0?41%, P<0?05;0.74± 0?16 vs. 0.68± 0?20, P<0?05);the position differences between two automatic delineation approaches were less than 0?4 cm in all three directions except for the temporal lobe, lower jaw, and spinal cord;in the receiver operating characteristic curve defined by Se versus 1-Sp , the data points were all within the first quadrant except for the optic nerve and chiasm;automatic delineation saved 42%?72% of time compared with manual delineation. Conclusions The ABAS software achieves satisfactory results of automatic delineation for most of OARs in patients with head and neck cancer. The multi?template automatic delineation, particularly, has better outcomes than the single?template one. In addition, it greatly shortens the time the clinicians spend on delineation of OARs.

8.
Article in Chinese | WPRIM | ID: wpr-502155

ABSTRACT

Objective To investigate association between gene mutations of surfactant protein C (SP-C) exon Ⅱ and neonatal respiratory distress syndrome(NRDS).Methods From January 2013 to December 2014 in Neonatal Intensive Care Unit of Bayi Children's Hospital Affiliated to Beijing Military General Hospital,a total of 90 cases with NRDS were selected as NRDS group,and they were grouped into 3 subgroups,30 cases in each subgroup.Clinical data were collected by observation figures.A total of 90 cases without NRDS as control group were matched with NRDS group as for the gestational age (GA),gender and birth weight(BW).The study samples were from the clinically diagnosed venous sampling 1 mL,and genomic DNA was extracted with routine technique.After the polymerase chain reaction (PCR) amplification of sequencing was performed.Chromas software and Vector NTI Advanc software were used for analysis,the results were compared with the normal SP-C sequence of GenBank.Results Heterozygous missense mutations included c.115G > T and c.139G > C existed in exon Ⅱ of SP-C gene,which changed protein sequence.GT genotype frequency(0.067) of c.115G >T mutation in NRDS group was higher than that(0) in control group (x2 =7.283,P < 0.05).With c.115 G > T gene mutations in patients with NRDS,Ⅲ-Ⅳ grade chest changed and mortality was higher than that of control group.GC genotype frequency (0.067) of c.139G > C mutation in NRDS group was higher than that(0) in control group (x2 =6.207,P < 0.05).Among NRDS group,64 cases of cesarean section patients,GC genotype frequencies were lower than GG genotype frequency (x2 =9.276,P < 0.01),55 cases of patients needed 3-4 PS pulmonary surfactant treatment,GC genotype frequencies were lower than GG genotype frequency (x2 =5.343,P < 0.05).Conclusions Two heterozygous mutations were located in SP-C gene of exon Ⅱ in Chinese Han population of this study.These mutations have never been reported before.c.115G > T of SP-C gene mutation is associated with pathogenesis and severity of NRDS and increases the risk of death in NRDS;while c.139G > C of SP-C gene mutation is associated with pathogenesis of NRDS,but it does not increase the risk of severity and death in NRDS.

9.
Article in Chinese | WPRIM | ID: wpr-478821

ABSTRACT

Acute kidney injury( AKI) is a variety of causes leading to rapid impaired renal function, causing glomerular filtration rate reduced,blood urea nitrogen elevated,serum creatinine increased,urine out-put decreased,resulting in the disorder of the human body environment,so the body can not maintain homeo-stasis. AKI increases the risk of mortality and chronic kidney disease in neonate. To understand the progress of AKI on diagnostic criteria,pathogenesis,early biomarker,treatment and prognosis,it is extremely important to guide clinical diagnosis and treatment,improve the outcome and reduce the mortality of AKI in neonate.

10.
China Pharmacist ; (12): 1770-1772, 2014.
Article in Chinese | WPRIM | ID: wpr-475740

ABSTRACT

Objective:To establish a method for the determination of asperosaponinⅥin Dieda Cuyu tablets by HPLC. Methods:A Hypersil C18(250 mm ×4.6 mm,5 μm)column was used. The mobile phase was acetonitrile-water(30∶70) with a flow rate of 1.0 ml·min-1 . The detection wavelength was 212 nm, the column temperature was room temperature,and the injection volume was 10μl. Results:AsperosaponinⅥ showed a good linear relationship within the range of 0. 04-0. 32 μg(r=0. 999 6). The average recovery was 97. 84%(RSD=1. 70%, n=6). Conclusion:The method is simple,accurate and reproducible, which can be used in the deter-mination of asperosaponinⅥ in Dieda Cuyu tablets.

11.
Article in Chinese | WPRIM | ID: wpr-444308

ABSTRACT

Objective To compare the static intensity-modulated radiotherapy (IMRT) and volumetric modulated arc therapy (VMAT) for mid and upper thoracic esophageal cancer.Method The data of twenty esophageal cancer patients were retrospectively re-planned with VMAT(single arc and double arcs) modality using Pinnacle treatment plan system.Five of these patients were selected again to simulate single arc plans with different segment intervals (4°,3°,2°) and re-planned on other treatment planning systems (Monaco and MasterPlan).Differences of dose distribution and treatment parameters were compared.Results In comparison to IMRT and single-VMAT (S-VMAT),Double-VMAT (D-VMAT) significantly improves the dosimetric parameters for targets(P < 0.05),dose homogeneity(P < 0.05) and conformity(P < 0.05).Though VMAT plans were slightly better than IMRT in reducing the doses to the organs at risk (OARs),no advantage was observed in the low-dose protection of lung and E-P (P < 0.05).For the VMAT plans with different segment intervals,lower OAR doses were observed using an interval of 2°(P < 0.05),except for the mean dose of the heart.For the VMAT plans on different treatment planning systems,Monaco-based plans protected OARs better (P < 0.05).The number of monitor units (MU) and treatment time were less in VMAT cases.Conclusions VMAT plans perform better in target coverage,dose homogeneity and conformity,and can reduce the radiation dose to the spinal cord,lungs,heart and other normal tissue than IMRT plans.The VMAT plan quality could be further improved by using double arcs and smaller segment interval.Monaco-based plans provide better OAR protections under the same conditions of physical and optimization parameters.

12.
Article in Chinese | WPRIM | ID: wpr-439169

ABSTRACT

Objective This study aimed to explore the clinical characteristics and treatment of congenital central hypoventilation syndrome(CCHS),to raise awareness on the CCHS for early diagnosis and treatment.Methods The clinical data of 4 babies with CCHS were analyzed,all of the babies were repeated bruising,hypercapnia,and difficult-to-wean.After the relevant checks,we precluded the primary diseases of heart,lung and neuromuscular dysfunction that lead to low ventilation,and detected the major pathogenic genes,Phox2b of CCHS,refer to literature and diagnostic criteria of CCHS.Results Four babies all had typical clinical features of CCHS:enough ventilation when awake.But when they were asleep,their respiratory rate slowed down,hypoventilation,cyanosis and hypercapnia appeared,and they can not be awaked by hypercapnia and hypoxemia caused by hypoventilation.Genetic testing confirmed the presence of Phox2b mutations.Two infants were supported by non-invasive biphasic positive airway pressure(BiPAP),an infant was discharged home at 3 months of age with non-invasive BiPAP,and an infant was discharged home at 1 month of age.Both of them were monitored and treated at home,and showed normal development.Conclusion For babies who were repeated bruising,hypercapnia,and difficult-to-wean,but no primary diseases of heart,lung and neuromuscular dysfunction that lead to low ventilation,CCHS must be considered.Genetic testing of Phox2b can be used as an important diagnostic tool,and non-invasive BiPAP is one of the efficacious methods in the treatment of CCHS.

13.
Article in Chinese | WPRIM | ID: wpr-427033

ABSTRACT

ObjectiveTo investigate whether the T1-weighted dynamic contrast-enhanced perfusion magnetic resonance imaging (DCEPMRI) technique can help to delineate the clinical target volume of brain glioma patients.MethodsThe DCE T1-weighted images from 28 glioma patients were collected after GdDTPA was injected.After the acquired images were processed and analyzed using modified Tofts-Kermode'two compartment analysis model and de-convolution method,the value and its pseudo mapping of quantitative parameter Ktrans related to microvascular permeability were obtained.The tumor size in the largest diameter slice measured both in routine enhanced MRI and Ktrans mapping of T1-weighted DCEPMRI were compared.ResultsThe vascular permeability and tumor infiltration was lower in low grade glioma,the difference of the tumor size between T1-weighted DCEPMRI and routine enhanced MRI reached 0.2% -0.3% there was significant difference of tumor size between T1 -weighted DCEPMRI and routine enhanced MRI ( grade Ⅰ and Ⅱ grade with 2.93 cm2∶2.46 cm2(t=6.90,P=0.000) and 4.18 cm2∶3.21 cm2(t=10.22,P=0.000) ).While in high grade glioma,the vascular permeability and the tumor infiltration were higher,the difference of the tumor size between T1-weighted DCEPMRI and routine enhanced MRI reached 25% - 26%( the size of grade Ⅲ and Ⅳ were 6.46 cm2 vs 5.48 cm2 ( t =10.83,P =0.000) and 8.26 cm2 vs 6.52 cm2(t =18.53,P =0.000) ).ConclusionsThe pseudo mapping of quantitative parameter Ktrans related to microvascular permeability acquired by T1-weighted DCEPMRI reflect the infiltrating circumscription in glioma,T1-weighted DCEPMRI can provide more information in delineation the clinical target volume,and it can be used as a new method for tumor volume evaluation.

14.
Article in Chinese | WPRIM | ID: wpr-418355

ABSTRACT

Objective To investigate the situation of oxygen supplement and the incidence and clinical characteristics of long-term oxygen inhalation newborns in neonatal intensive care unit(NICU).Methods The records of oxygen supplement and the related clinical data of 12 155 neonates admitted in our NICU from Oct 2009 to May 2011 were collected and retrospectively analyzed.The results were compared with the data from a survey on 19 hospitals in China which reported by other authors.Results In 12 155 newborns,4 951 were full term,7 204 were preterm.One hundred and two patients (0.84%,102/12 155 ) accepted oxygen for more than 28 days.Among them,88 were preterm,14 were full term,with the average gestational age (31.16 ±3.70) weeks,the average birth weight (1.60 ±0.68) kg and the mean oxygen supplement period (40.60 ± 12.25) d.Finally,98 were cured or improved,4 died.The incidence of bronchopulmonary dysplasia (BPD) in 7 204 preterm infants was 1.22% ( 88/7 204) according to the standard of continuous oxygen supply more than 28 days after birth.The incidence of BPD in preterm infants less than 32 weeks was 4.92% (68/1 381 ) according to the standard of continuous oxygen supply more than 28 days after birth,while the rate was only 2.10% (29/1 381 ) according to the standard of continuous oxygen supply more than 36 weeks postmenstrual age.The rates of BPD according to the two different standards were significantly different ( x2 =16.251,P <0.001 ).There were significant differences in the rate of supply oxygen( x2 =119.99) and supply oxygen time( F =109.27 ) among different gestational age groups in overall the 5 499 neonates ( P <0.001 ),but no significant differences in the average time of oxygen supply and mechanical ventilation among different gestational age groups in infants with long-term oxygen dependence ( P > 0.05 ).There were significant differences in rates of pulmonary surfactant therapy,heart failure,retinopathy of prematurity,congenital heart disease,other congenital malformation and mortality among different gestational age groups in long-term oxygen dependence infants (x2 =8.789,13.538,23.176,7.778,8.842,8.246,P < 0.05 ).As compared with the data from 19 hospitals,the corrected rate of long-term oxygen supplement in preterm infants in our hospital was obviously lower[0.99% (71/7204) vsl.54% (190/12 351),P <0.001].Conclusion Theincidence of BPD in our NICU is low.Lower gestational age,immature lung and secondary lung injury may be the mainly cause of neonatal long-term oxygen dependence,but some factors such as congenital heart disease,congenital malformations should be considered in more mature infants.The most appropriate standard for BPD still remains to be discussed.

15.
Article in Chinese | WPRIM | ID: wpr-472037

ABSTRACT

Objective To assess the impact of different number of diffusion gradient directions (NDGD) of diffusion tensor imaging (DTI) on dispersion degree of fractional anisotropy (FA) values and signal-to-noise ratio (SNR) for adult brain tissues. Methods Eight healthy volunteers underwent DTI with 7 NDGD respectively, and the associated FA maps were obtained. Four region of interest (ROI) (genu and splenial of corpus callosum, genu and posterior limb of internal capsule) were chosen in white matter of FA maps, FA values and the corresponding SNRs of the ROIs were computed and analyzed with NDGD increasing. Results FA values within all ROIs did not change remarkably with NDGD increasing. Variance of FA value within two ROIs with high gray value (genu and splenial of corpus callosum) fluctuated randomly and SNR increased slightly, while variance of FA value within the ROIs with low gray value (genu and posterior limb of internal capsule) diminished significantly with NDGD increasing. Conclusion In clinical applications of DTI, an optimum NDGD for DTI data acquisition should be selected according to the ROIs in human brain to be inspected.

16.
Journal of Clinical Pediatrics ; (12): 220-222,250, 2010.
Article in Chinese | WPRIM | ID: wpr-583463

ABSTRACT

Objective To study the role of deferoxamine(DFO)on regulating hypoxia-inducible factor 1α(HIF-1α)expression after hypoxia-ischemia brain damage(HIBD)in neonatal rats,to explore the therapeutic strategy for HIBD. Methods Postnatal day 10 SD rats were divided into four groups: hypoxia-ischemia(HI)group,DFO-treated group,normal saline(NS)-treated group,and sham operation group. HIBD model was established by the ligation of right common carotid artery following the inhalation of nitrogen-oxygen mixtures containing 92% nitrogen and 8% oxygen. DFO-treated group and NS-treated group were treated by intraperitoneal injection of DFO or NS respectively. The brains were collected at 4 h,8 h,and 24 h after hypoxia. HIF-1α protein expression was detected by Western blot analysis,and HIF-1α mRNA expression was detected by using RT-PCR at each time point. Results The synthetic level of HIF-1α protein increased significantly at 4 h,peaked at 8 h,and decreased at 24 h after HI in HI group,as well as NS-treated group. However,in DFO-treated group HIF-1α protein was peaked at 4 h,maintained higher level at 8 h and 24 h after HI. The level of HIF-1α protein was much higher in HI and DFO-treated groups than those in sham controls(P < 0.05). The synthetic level of HIF-1α protein were higher in DFO-treated groups than those in HI groups at each time point(P < 0.05). HIF-1α mRNA expression was higher in DFO-treated groups than those in HI groups at each time point. Conclusions DFO upregulate HIF-1α protein and mRNA expression in neonatal rats with HIBD. The peak of HIF-1α protein expression are also more advanced and lasted longer after DFO-treatment.

17.
Article in Chinese | WPRIM | ID: wpr-408700

ABSTRACT

BACKGROUND: Pleiotropia and indeterminateness of the differentiation of neural stem cells (NSCs) cause great difficulties for clinical application.Therefore,it is the key to solving the problem to investigate the proliferation and differentiation condition of NSCs.OBJECTIVE:To observe the effects of epidermal growth factor(EGF)and basic fibroblast growth factor (bFGF) on the proliferation and differentiation of neural stem cells of human embryonic brain.DESIGN: A randomised and controlled experiment taking cultured human embryonic stem cells as subjects.SETTING: Department of Pediatrics of Zhujiang Hospital, the First Military Medical University of Chinese PLA.MATERIALS:This experiment was conducted at the Central Laboratory of the Department of Pediatrics of Chinese PLA from January to May 2004.Two 16-week embryos from induced labor by water bags voluntarily were chosen at random from the Department of Obstetrics in a tertiary hospital in Guangzhou(Informed consent was obtained from the parents of the fetuses).Then,cells in the subventricular zone were cultured in serum-free medium and serum medium, respectively.METHODS:Primary cells of subventricular zone of human embryonic brain were cultured with serum-free nedium containing EGF,bFGF and EGF + bFGF.The concentration of two growth factors was both 20 μg/L;experiment of differentiation was performed on the cell neurospheres cultured from the primary generation with serum culture medium;differentia tive cells were detected with immonohistochemical technique.MAIN OUTCOME MEASURES:The number of neurospheres and the change of neurons and gliocytes from neurospheres in each group.RESULTS:① There was no significant difference in the number of primary neurospheres between bFGF group and EGF+bFGF group [(150.3±14.9) /well vs (173.6±26.4)/we11, P > 0.05] , but the number in the two groups was both more than that in EGF group [(99.5±14.9)/we11, P < 0.01].② The neurons differentiated from neurospheres in bFGF group and EGF+bFGF group outnumbered those in EGF group, but gliocytes in EGF group outnumbered those in EGF + bFGF group.CONCLUSION:① bFGF can promote the proliferation of neural stem cells of subventricular zone of human embryonic brain,and the formed neurospheres can differentiate more neurons.② Combination of EGF and bFGF is not superior to single bFGF in effect,suggesting that there is no synergistic effect.

18.
Article in Chinese | WPRIM | ID: wpr-409456

ABSTRACT

BACKGROUND: Neural stem cells(NSCs) have been used to treat brain injury or some degenerating diseases of nervous system. Since in vitro culture conditions for NSCs differ from normal physiological conditions, whether the properties of the cultured cells are consistent with those of cells under physiological conditions? Therefore, inducing endogenous NSCs to proliferate and differentiate may be more promising for practise of NSCs.OBJECTIVE: This study was designed to investigate the developmental properties of NSCs in frontal cortex of embryonic human brain at various ages.DESIGN: It was a randomized experimental study.SETTING: This study was conducted at Department of Pediatrics, Zhujiang Hospital, Southern Medical University.PARTICIPANTS: Totally 90 16-to-36-week-old fetuses underwent inducing abortion by water bag were selected at the Obstetrics & Gynecology Department of Zhujiang Hospital Affiliated to Southern Medical University from October 2003 to March 2004. Brain tissue was taken from the frontal cortex of the aborted fetuses. All the mothers had normal physical examination findings. The informed consents on inducing abortion by water bag had been obtained from relatives and the mothers. The study was conducted with a prior permission from the competent department of the First Military Medical University. According to their ages, the fetuses were divided into 6 groups,16-week group, 20-week group, 24-week group, 28-week group and 36-week group, each group containing 15 cases.METHODS: After inducing abortion by water bag, under axenic conditions, the aborted fetus was dissected, with the scalp excisd, the skull opened and the membrane covering brain pull apart. Then the frontal cerebral cortex was taken out, fixed and sliced. Employing immunohistochemical staining and light microscope, distribution, morphological features, phenotypes, growth patterns and quantity of NSCs in the frontal cortex were observed. Morphological features of the cells and expressions of markers in the cells were examined under light microscope. Negative control was set according to the substitution method. Under a × 400 field of microscope, some nestin-positive cells with speckled brown cytoplasmic staining were defined as NSCs. Two slides of each sample were detected and 10 fields of each slide were observed. Based on these observations, in each group, the total number of cells and the number of positive-stained cells in 300 fields were counted. The rates of nestin-positive cells were calculated.MAIN OUTCOME MEASURES: Morphological features, quantitative assessment and developmental features of the nestin-positive NSCs in frontal cortex of embryonic human brain at various ages were main outcome measurements in this study.RESULTS: NSCs were found in frontal cortex of embryonic human brain. They mainly were distributed in the pyramidal layer and the internal granular layer. They were small round-or oval-shaped, most were small round-shaped. These cells had a relatively large vacuolar nucleus with 1 - 3 nucleoli, loose chromatin and marked cytoplasmic staining. Some of the round-shaped cells were mitral cells with short neurites. The oval-shaped cells had 2 neurites. A distinct territorial distribution of NSCs could be observed. Some colonies, consisting of a few NSCs and looked like the neurospheres in in vitro culture, could be seen. Occasionally, symmetrical division of NSCs could be observed. In all the groups, 16-week, 20-week, 24-week,28-week, 32-week and 36-week group, the rates of nestin-positive NSCs in frontal cortex of embryonic human brain decreased with the increase of age (15.59%, 13.48%, 11.62%, 10.52% ,9.87% ,6.68% ,X2 = 1 265. 152, P< 0.01).CONCLUSION: The distribution, morphological features, phenotypes, growth pattern and quantity of the NSCs in frontal cortex of embryonic human brain at various ages are different and auto-developmental features exist. The number of these cells decreases with the increase of age.

19.
Article in Chinese | WPRIM | ID: wpr-587831

ABSTRACT

Objective:To investigate the changing characteristics of neural stem cells(NSC) in the temporal lobe of cortex from human fetal brain at different age.Methods:Ninety embryos delivered by induction labor with water bag at gestational age 16-36 w were collected,distribution,Shapes,growth modes and the number of NSC in the temporal lobe of cortex were determined with immunohistochemical method under light microscope. Results:NSC existed in the temporal lobe of cortex from human fetal brain and were round,elliptic or polygonal in shape,especially small round,NSC distributed mainly in cone cell and inner granule cell layers.NSC had enations from 0 to 2,nucleoli from 1 to 2 and rarefactive chromatin,nucli were large.Most NSC distributed in clones with a few NSC existed in a single mode among other nerve cells.There were no differences between groups in the distribution,shapes,species and growth modes of NSC in the temporal lobe of cortex,but the numbers of NSC gradually decreased with increase of age.Conclusion:There were no differences on the distribution,shapes,species and growth modes of NSCs in the temporal lobe of cortex from different gestational age,the number of NSC decreased with increase of age.

20.
Article in Chinese | WPRIM | ID: wpr-555742

ABSTRACT

Objective To study the development of neural stem cells from human fetal brains at different developmental stages. Methods A total of 100 cases of embryos at 16-32 gestational weeks by induction of labor with water bag were collected for the determination of the distribution, forms, existing modes, and the number of neural stem cells in the hippocampus by SABC immunohistochemical method and light microscopy. Results Neural stem cells were found in the hippocampus at different fetal ages and located in the polymorphic layer, pyramidal, granular and molecular layers of hippocampus, mainly in polymorphic layer, pyramidal layer, and granular layer. Neural stem cells in hippocampus were round, ellipse, triangle, and stellate, particularly round and ellipse. No obvious enation was found. Neural stem cells had plenty of cytoplasm. The nuclei were round and ellipse with rare chromatin and nucleoli from 2 to 6. Most of neural stem cells were distributed among other neurons, and symmetric cleavage was found in some of them, but some neural stem cells were distributed in cluster and nest. The number of neural stem cells in hippocampus were different between groups and gradually decreased with the increasing gestational age. Conclusion Neural stem cells exist widely in the hippocampus at different gestational ages. There are differences in distribution, forms, existing modes, and number of neural stem cells in hippocampus at different gestational ages. Hippocampus may be the new originating region of neural stem cells.

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