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: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.

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.

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.

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.

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.

Objective To discuss the influence of intravenous anesthesia with procaine on the release of central nervous Ach. Methods 50 samples of plasma were collected from healthy people, and 30 samples of plasma from cases prior to surgical operation, during anesthesia and after operation respectively. The activity of Ach in the plasma was measured. Then observe the dropping speed and concentration of procaine at different time during anesthesia, and the subsequent changes of the activity of plasma Ach and the slowdown of its activity. Results The data showed that the activity of plasma Ach prior to operation was considerably lower than that of the control group (P

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.

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.

Objective To compare the clinical outcomes between two different femur rotating osteotomy methods for kneeling ability recovery after total knee arthroplasty (TKA).Methods From January 2012 to December 2014,88 patients underwent TKA were selected for a retrospective study and were divided into two groups based on the methods to determine femoral rotation.Forty-eight patients were in measured resection group,while 40 patients in gap balancing group.The patients in both groups underwent fixed-bearing tibia prosthesis.There were no statistical significance between the two groups in gender,age,BMI and knee varus angle (P＞0.05).The knee varus angle,ROM,Oxford knee score (OKS) and American Knee Society (KSS) knee score were collected to assess malformation correction,kneeling ability and functions at pre-operation,one and two years postoperatively.Results The operation duration and blood loss in measured resection group were 80±19 min and 348±121 ml,while these data in gap balancing group were 82±23 min and 315 ± 100 ml respectively (P＞0.05).Patients in measured resection group were followed up 24-59 months (mean 43± 11 months),while the followed-up duration in gap balancing group was 25-58 months (mean 47±10 months).No major complications such as infection loosen and instability were occurred.Varus angles in measured resection group at postoperative 1 year and 2 year postoperative were 1.2°±0.4° and 1.0°±0.2° respectively,while those in gap balancing group were 0.9°±0.2° and 0.8°±0.3° (P＞0.05).The scores of the seventh item of OKS in measured resection group at 1 year and 2year follow-ups were 2.79±1.02 and 2.75± 1.03 respectively,while those in the gap balancing group were 1.90±0.85 and 1.80±0.83 (P＜0.01).ROM in the measured resection group at 1 year and 2 year postoperative were 102.08°± 15.60° and 102.08°±15.60° respectively,while those in the gap balancing group were 112.50°±18.32° and 113.00°±18.09° (P＜0.05).KSS in measured resection group at postoperative 1 year and 2 years were 154.63±31.12 and 154.63±31.26 respectively,while those in the gap balancing group were 170.55±22.67 and 173.45±22.52 (P＜0.05).Conclusion The method of measured resection and gap balancing to confirm femoral rotation during TKA can both achieve favorable kneeling ability and clinical outcomes,while gap balancing show superiority on kneeling ability recovery,ROM and clinical outcomes at 2-year postoperative improvement.