BACKGROUND: How to acquire high-purity epidermal stem cells (ESCs) and maintain ESC phenotype and in vitro proliferation characteristics are urgently solved problems in the in vitro culture of ESCs.OBJECTIVE: To establish methods to in vitro isolate and culture human ESCs.DESIGN,TIME AND SETTING: Cell observation was performed in the Department of Bum,First Affiliated Hospital of Nanchang University.MATERIALS: Foreskin was obtained front 2-12-year-old patients who were subjected to circumcision.These patients did not suffer from urinary system infection.METHODS: Keratinocytes and fibroblasts were isolated from foreskin using Dispase Ⅱ and trypsin.ESCs were isolated by rapidly adhering to type Ⅳ collagen.The supematant fluid of fibroblasts of passages 2-3,which were in the exponential growth phase,was collected.After screening,the supematant fluid was mixed with serum-free medium of keratinocytes at the proportion of 1:1.Simultaneously,fetal bovine serum,epidermal growth factor,and bovine pituitary extract were added to prepare medium of ESCs,which was used for in vitro proliferation of human ESCs.Homologous keratinocytes were used as controls.MAIN OUTCOME MEASURES: After 2 passages,cloning efficiency and cloning sustaining time were calculated by plate clone formation assay.Expression levels of β 1 integrin and keratin 19 were detected by immunohistochemistry.RESULTS: The cloning efficiency of human ESCs was significantly increased,and the cloning sustaining time was significantly prolonged as compared with homologous keratinocytes cultured in the same batch (P ＜ 0.01).lmmunohistochemistry results demonstrated that both β 1 integrin and keratin 19 were positive in the human ESCs.CONCLUSION: Keratinocytes could be successfully isolated by means of rapid adherence to type Ⅳ collagen and cultured with conditioned medium in vitro.

BACKGROUND:Stem cel s have the potential to differentiate into various organs and tissues. In recent years, stem cel s have been proved to differentiate into hair fol icles under certain conditions. OBJECTIVE:To review the research progress and prospect of stem cel s differentiating to hair fol icles, thereby providing a reliable basis for clinical treatment of serious hair fol icle injury. METHODS:A computer-based search of PubMed, EMBASE, WanFang, CNKI databases was performed for related articles published between 2013 and 2015, using the keywords of“cel hair, fol icle stem, medicine regenerative, differentiation”in English and Chinese. A total of 207 articles were retrieved, and final y 34 articles were included in result analysis. RESULTS AND CONCLUSION:Stem cel s from different sources al have the ability to differentiate into hair fol icles under certain inductions. However, it is important to seek more scientific and rational methods for the differentiation of stem cel s into hair fol icles based on overcoming their own shortcomings. A great progress has been made in animal experiments and subclinical trials, and even a great breakthrough in some aspects. Further studies on combining the advantages and overcoming the shortcomings of various stem cel s during differentiation are required for the clinical treatment of serious hair fol icle injury.

Objective To investigate the mutation characteristics of atlastin gene in Chinese patients with hereditary spastic paraplegia(HSP) and establish the base of gene diagnosis of HSP.Methods Mutation analysis of atlastin gene was made by use of polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) combined with DNA direct sequencing in 30 unrelated affected HSP individuals in China in which 20 cases were from autosomal dominant families and ten cases were sporadic HSP patients.Results No abnormal SSCP bands were found in the 30 individuals and the results of DNA direct sequencing were also normal.Conclusion Mutation of atlastin gene may be rare in Chinese HSP patients.

Objective To investigate the mutation characteristics of paraplegin gene in Chinese patients with hereditary spastic paraplegia (HSP) and establish the base of the gene diagnosis of HSP.Methods Mutation analysis of paraplegin gene was carried out by use of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with DNA direct sequencing in 22 unrelated affected HSP individuals in China, in which 8 probands were from autosomal recessive families and 14 cases were sporadic.Results All of the exons could be detected by PCR. 2 probands were found to have abnormal SSCP bands in exon 15 and 2 substitutes (G2063A, G2066A in exon 15) were found by DNA direct sequencing. But there were no changes in other patients of families.!The same abnormal SSCP bands and G→A substitutes were revealed in control individuals. So these changes were two polymorphisms, in which G2066A was not reported previously.Conclusion Mutations of paraplegin gene may be rare in Chinese patients with HSP. G2063A and G2066A are two polymorphisms, in which G2066A has not been reported previously.

BACKGROUND:Ideal dural substitute materials requires its good biomechanics properties,tenacity and elasticity.OBJECTIVE:To determine freeze-dried and irradiated porker dura's resistant ability,including type Ⅰ collagenase digestion,the biggest load and tensile strength,to assess the porker's dura irradiated and freeze-dried whether or not can satisfy the biomechanics requirment in dura draft.DESIGN,TIME AND SETTING:This controlled analysis experiment was performed at the Burn Institute,Nanchang University,Association Mechanics Laboratory,Nanchang University-New Sans Company,Chinese Crude Drug Solid Preparation Nation Project Research Center of Jiangxi Traditional Chinese Medical College and Jiangxi Tianzhao Technology Development Company from July 2007 to June 2008.MATERIALS:Six healthy living pigs was used to sterilely obtain fresh dura mater.METHODS:After washing,pre-frozen,freeze-dried and radiation by Co ?-ray,we prepared fresh,freeze-dried,irradiated and 60 freeze-dried plus irradiated pig dura.MAIN OUTCOME MEASURES:5-mm wafer of four group dura were dealt with typeⅠcollagenase meanwhile,digested time was recorded.EMT color screen electronic testing machine was used to measure the biggest load and tensile strength of the porker dura in each group.RESULTS:Time of the dura resistance to collagenase digestion were respectively(8.3 ? 2.5),(7.6 ? 1.8),(23.6 ? 5.7)and(21.1 ? 5.3)minutes,and the largest load and tensile strength were(79.93 ? 4.36),(70.50 ? 5.97),(96.97 ? 4.84)N and(93.59 ? 4.61),(7.98?0.44),(7.05?0.60),(9.70?0.48)and(9.40?0.46)N/mm in the fresh,freeze-dried,irradiated,and freeze-dried plus irradiated pig dura groups.Compared with the fresh and freeze-dried pig dura groups,the time was longer in the irradiated and freeze-dried plus irradiated pig dura groups(P 0.05).CONCLUSION:Dural substitute materials irradiated by Co ?-ray enhanced the ability against enzymatic hydrolysis and 60 biomechanics.Irradiated and freeze-dried plus irradiated dural substitution can meet the requirements of the biomechanics in dura mater transplantation.

BACKGROUND: Wuhuang oil has a bacteriostatic action to treat burn wound and promote traumatic healing, but the action on inhibition of scars formation remains poorly understood.OBJECTIVE: To investigate the effects of modified Wuhuang oil at different concentrations and administration times on the growth and proliferation of human fibroblasts in vitro.DESIGN, TIME AND SETTING: Comparison observation regarding cytology in vitro was performed at the Burns Institute in the First Affiliated Hospital of Nanchang University between April 2006 and January 2007.MATERIALS: Prepuce specimens were harvested from patients who underwent circumcision in Department of urinary surgery, at First Affiliated Hospital of Nanchang University and Jiangxi Provincial Children Hospital. All patients aged 2-12 years old, and informed consents were obtained from their relatives. Wuhuang oil and modified Wuhuang oil (water-solubility) were offered by Department of Pharmaceutical Preparation in the First Affiliated Hospital of Nanchang University, China. METHODS: Human fibroblasts cultured in vitro were divided into 2 groups at random, experiment and control. Experiment group was treated with 300 g/L Wuhuang oil, while control group with 300 g/L modified Wuhuang oil. Serum-free culture fluid was used to prepare 6 concentrations of oil solution: 0 (blank control), 100, 150, 200, 250, 300 g/L.MAIN OUTCOME MEASURES: MTT assay was used to determine the growth and proliferation of human fibroblasts at 2, 3, 4, 5,6 days; inhibition rate of cell growth was observed at 2, 4, 6, 8, 10 days.RESULTS: Modified Wuhuang oil (0-300 g/L) concentration positively correlated with inhibition of human fibroblast proliferation;the inhibition was not related to culture time. Modified Wuhuang oil (300 g/L) had the greatest inhibition rate of human fibroblasts at 8-10 days, there were significant differences between experiment group and control group (P < 0.01).CONCLUSION: Modified Wuhuang oil has an effective inhibition on the proliferation of human fibroblasts in vitro, and shows a dose-dependent tendency. Compared with Wuhuang oil, 300 g/L modified Wuhuang oil is superior to suppress the growth of human fibroblasts.

OBJECTIVE:To establish a method for the dissolution determination of Dronedarone hydrochloride tablet,and eval-uate the quality consistency of its generic and original preparations. METHODS:UV spectrometry was performed on the column of 288 nm,dissolution media of Phosphate buffer solution (pH4.5),0.1 mol/L Hydrochloric acid solution [adding into 0.5% sodium dodecyl sulfate(SDS)],Phosphate buffer solution [pH6.8,adding into 0.5%SDS] and water,volume of dissolution medium was 1 000 ml,rotation speed was 75 r/min,the dissolution of generic and original preparations of Dronedarone hydrochloride tablet was detected,and the similarity of dissolution curve was evaluated by calculating the similarity factor (f2). RESULTS:The linear range of dronedarone hydrochloride was 2.147-25.764 μg/ml;RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries 4 dissolution media were 99.53%-101.05%(RSD=0.48%,n=9),98.95%-100.05%(RSD=0.39%,n=9), 99.54%-100.20%(RSD=0.24%,n=9)and 98.54%-100.06%(RSD=0.44%,n=9). In the 4 dissolution media,f2 of the dissolu-tion curve of 3 batches of generic and original preparations of Dronedarone hydrochloride tablet was 56,60,63,68,68,52,59, 67,65,68,76,62,respectively. CONCLUSIONS:The method is suitable for the dissolution determination of Dronedarone hydro-chloride tablet;meanwhile,the in vitro dissolution curves of generic and original preparations of Dronedarone hydrochloride tablet show similarity,so the quality consistency is good.

BACKGROUND:Cerebral vascular malformations are the leading cause of hemorrhagic apoplexy in young adults, and the rupture and bleeding of malformed vessels may cause severe neurological dysfunction. The mechanism of cerebral vascular malformations remains unclear. Modern molecular biology studies have shown that, angiogenic growth factors are abnormal y expressed in cerebral vascular malformations.
OBJECTIVE:To evaluate differences in the expression of angiogenic growth factors in cerebral vascular malformations, and discuss the possible relationship between cerebral vascular malformations and angiogenic growth factors.
METHODS:Fifty patients with cerebral vascular malformations and fifty patients with intracerebral hemorrhage were included in this study. The expressions of angiogenic growth factor (vascular endothelial growth factor and transforming growth factor-α) in the cerebral vascular malformation specimens and the normal superficial temporal artery specimens were detected with immunohistochemical staining.
RESULTS AND CONCLUSION:In the normal superficial temporal artery of intracerebral hemorrhage patients, no expression of vascular endothelial growth factor and transforming growth factor-αwas found;in the vascular malformations, they were highly expressed (P<0.05). Compared with normal blood vessels, vascular endothelial growth factor and transforming growth factor-αexpression was significantly increased in patients with cerebral vascular malformations.

Furthering of the health reform calls for better health care system,greater public financing,and deepened reform of public hospitals.Introduced in this paper are the actions taken and outcomes of the health reform in Changchun city,including the essential medicines system to minimize drug prices,improvement of the primary healthcare system to minimize people's financial burden,greater public spending to enhance public welfare nature of medical institutions,enhanced competencies of primary medical institutions to introduce rational classification of patient flow,and enhanced equity of public services to promote people's health.On such basis,further reform proposals were made for Changchun's health reform.

Over-expression of APP and Swedish mutation could cause some familial early onset AD. In this study, a primary screening was conducted of effective small interference RNAs (siRNAs) targeted wild type APP (APPwt) and Swedish mutant APP (APPswe). One siRNA targeting APPwt and the other siRNA targeting APPswe were designed. All these siRNAs were endogenously expressed by siRNAs expressing plasmids. COS-7 cells were transiently co-transfected with APP-GFP recombinant plasmids and siRNA expression vector. The silencing effect of each siRNA was quantitatively assessed by the level of expression of green fluorescent protein (GFP). It was found that the siRNAs silenced APPwt and APPswe to different degrees. siRNA directed against APPswe was more effective in suppressing the expression of fusion gene of APPswe than that of APPwt. The silencing effect of siRNA directed against APPswe indicating allele-specific silencing property of the siRNAs. Therefore, siRNAs directed against APP play an important role both in the therapeutic study of Alzheimer disease and functional exploration ofAPP gene.