Objective To investigate the therapeutic effect of transplantation of limbal epithelial cells and decellularized porcine cornea construct in rabbits with corneal alkali burn.Methods Allogeneic rabbit limbal epithelial cells were cultured and identified.Porcine cornea scaffold was fabricated by decellularization.The LECs-DPC construct was cultured and transplanted to the alkali-burned corneal surface of rabbits(4month follow-up).Results Immunocytochemistry and RT-PCR analyses revealed the presence of ABCG2 positive limbal stem cells.A general view showed the corneas were clear and vessel-free postoperatively.HE and Masson Trichrome staining results indicated the epithelial stratification,fewer cellular infiltration,DPC' integration into host corneal tissues,and more regular collagen configuration.Transmission electron microscopy exhibited formation of desmosomes and microvilli.Conclusion Allogeneic limbal epithelial cells transplantation using DPC as a carrier can restore corneal clarity after alkali burn in a rabbit model.

Objective To investigate the diagnostic value of the early slope value of enhancement in differentiating benign from malignant musculoskeletal masses.Methods Dynamic contrast-enhanced gradient-recalled acquisition in the steady state imaging(GRASS) was performed to evaluate 36 musculoskeletal masses(18 malignant,18 benign).The difference of the early slope value of enhancement was analysed and compared between benign lesions and malignant ones.Using ROC curve plotting obtained the best decisive threshold.Spearman rank correlation analysis was used to determine the relation between slope and microvessel density of tumor tissues.Results The average early slope value of enhancement was (4.33?2.70)% per second (range 0.97%～10.50% per second)for malignant lesions,and was (1.46?0.97)% per second(range 0.37%～3.86% per second)for benign ones.There was an apparent statistical significance between benign and malignant tumors(?

Objective To investigate the correlation of dynamic contrast-enhanced (DCE) MRI features with microvessel density (MVD) and vascular endothelial growth factor (VEGF) in non-small cell lung cancer(NSCLC).Methods Conventional MR imaging and dynamic contrast-enhanced scan in thirty-three patients with NSCLC confirmed by pathologyn were performed. MVD and VEGF were stained with immuno-histochemical technique in all cases. Some parameters of DCE MRI, including maximum slope(Smax) and time to peak(TTP) were put more analysis. The relationship between the results of DCE MRI (Smax and TTP) and that of immuno-histochemistry (MVD and VEGF) was analysed.Results The Smax of adeno carcinoma was higher than that of squamous cell carcinoma,but TTP was lower. The difference was obvious difference(t=3.22,P

BACKGROUND: Many scholars have paid attentions on neural stem cells (NSCs), which are regarded as the cell source to repair the injured nervous tissues. And it is becoming a key point that NSCs will be obtained in an effective way.OBJECTIVE: To observe the characteristics of isolation, proliferation and differentiation of in vitro NSCs from subventricular zone (SVZ) and hippocampus of neonatal rats.DESIGN: Single-sample trial.SETTING: Department of Neurosurgery in the Third Affiliated Hospital of Sun Yat-sen University.MATERIALS: Ten SD rats aged 3 days, of either gender, were offered from the Experimental Animal Center of Sun Yat-sen University. Nestin antibody (rabbit-anti-rat), neurofilament (NF-200) antibody (rabbit-anti-rat), and glial fibrillary acidic protein (GFAP) antibody (mice-anti-rat) were all purchased from Sigma Company.METHODS: The experiment was carried out in the Laboratory of Histology and Embryology, Basic Medical College of Sun Yat-sen University from September to December in 2006. According to the ethical requirement, the serum-free DMEM/F12 medium containing basic fibroblast growth factor and epidermal growth factor was utilized to isolate and incubate NSCs from SVZ and hippocampus of neonatal rats. Then the proliferation and differentiation of NSCs in vitro were observed.MAIN OUTCOME MEASURES: Fluorescence immunocytochemistry was applied to detect the antigen of nestin expressed from NSCs, NF expressed from neuron cells and GFAP expressed from astrocytes. The differentiation of NSCs was identified.RESULTS: The cells isolated from SVZ and hippocampus of neonatal rats possessed the ability of proliferation and self-renewal. Through inverted microscope, nestin positive cells could be found in the cell clone sphere at passage 20 of cells. Cells induced to differentiate could express antigen of NF and GFAP by the attached cell clone sphere.CONCLUSION: The NSCs successfully isolated from SVZ and hippocampi of neonatal rats possess the ability of self-renewal, proliferation, and differentiation into neuron-like cells and astrocytes.

Objective To observe the effect of MAC of the isoflurane at the general combined with epidural anesthesia with lidocaine and dicaine.Methods 78 cases of the cholecystectomy patients randomly double-blindly divided Ⅳ groups. General combined with epidural anesthesia were performed at the Ⅰ、Ⅱ、 Ⅲ groups. And general anesthesia were performed at the contronal group. The MAC of the isoflurane are checked with up-and-down method with electronic stimulation at left on C5 leveral. Results The MAC of the three observed groups of the general combined with epidural anaesthesia were 0.67?0.097%、0.68?0.084%、0.61?0.103 % differently. There is no siginificant difference.But the MAC of the general contronl group is 1.15?0.088%.There are siginificant difference with the three observed groups,The MAC of the isoflurane can be reduced with general combined with epidural anesthesia. Coinclousions The MAC the isoflurane can be reduced with general combined with epidural anaesthsia significantly compared with singal general anesthesia. And there are no significant different in the three observed groups.It is the pulsing action with lidocaine combined with dicaine at epidural anesthesia. The mechanism of anaesthesia with lidocaine or dicaine supposed to be the same.

Objective To explore the most important risk factors in metabolic syndrome(MS) components.Methods Ninety-four individuals were classified into MS and non-MS group according to the diagnostic criteria for MS proposed by Chinese Diabetes Society(CDS) revised in 2006 or International Diabetes Federal(IDF) in 2005.Age,waist circumference(WC),body mass index(BMI),fasting plasma glucose,lipid profile,blood pressure and blood cell counts in two groups were compared.Partial least squares discriminant analysis(PLSDA) was carried out to determine the most important components of MS.Results Patients with MS diagnosed by CDS or IDF criteria have significantly older age,higher BMI,WC,blood pressure,fasting plasma glucose,triglycerides,insulin levels,insulin resistance index,high sensitivity CRP and fibrinogen levels compared with non-MS group.PLSDA analysis shows WC,BMI,blood pressure and aging are most important components of MS.Conclusion Obesity,hypertension and aging are three most important components of MS with obesity is the utmost among them.

Eleven type 1 diabetic patients who received fixed regime of insulin Glargine were included in the study.The levels of fasting serum insulin were measured for each subject at 6:00 in three consecutive mornings.The variability of mean fasting serum insulin in each subject was 3.3%-41.5% (mean 15.4%).The variability did not correlate with the dose of Glargine statistically.

BACKGROUND:Inflammatory factor plays an important role in restenosis after bal oon injury. Sphingosine1-phosphate receptor 1 can enhance the expression of inflammatory factor and promote development and progression of this pathological process.
OBJECTIVE:To observe the expression of the inflammatory factors and sphingosine1-phosphate receptor 1 after bal oon injury of the rat carotid artery and effects of fingolimod hydrochloride on reducing inflammatory reaction.
METHODS:Sixty Sprague-Dawley rats were equal y and randomly divided into four groups. In the blank control group and negative control group, left common carotid artery was only isolated, and left external carotid artery was ligated. In the bal oon injury group and drug intervention group, rat models of carotid artery injury were
established by bal oon injury on the left common carotid artery. In the negative control and drug intervention groups, the rats were intraperitoneal y injected with fingolimod hydrochloride 1 mg/kg. In the blank control and bal oon injury groups, the rats were intraperitoneal y injected with an equal volume of saline. Samples were col ected at 3, 7 and 21 days.
RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that the proliferation of blood vessel was remarkable in the bal oon injury group, but attenuated in the drug intervention group. The appearance of blood vessels was normal in the blank control group and negative control group. Real-time fluorescent quantitative PCR revealed that cyclooxygenase 2 and prostaglandin E2 mRNA expression levels were significantly lower in the drug intervention group than those in the bal oon injury group at 7 days (P<0.05). Cyclooxygenase 2 and prostaglandin E2 mRNA expression levels were significantly higher in the bal oon injury group and drug intervention group than those in the blank control group and negative control group at the same time point (P<0.05). Western blot assay results revealed that sphingosine1-phosphate receptor 1 expression was high in early stage of injury, and then reduced in late stage of injury. In particular, protein expression further decreased after drug intervention. Results indicated that fingolimod hydrochloride suppressed inflammatory reaction of injured blood vessels and lessened the stenosis of injured blood vessels by regulating cyclooxygenase 2 and prostaglandin E2 mRNA expression using sphingosine1-phosphate receptor 1.

OBJECTIVE To investigate whether the KCNN4 gene and KPTN gene contribute to a Chinese non-syndromic hearing loss pedigree linked to DFNA4 with positional candidate approach. METHODS The complete coding region of the two genes were amplified with polymerase chain reaction (PCR), and bidirectional sequencing of the PCR products was subsequently applied in the 36 family members to identify the possible mutations or polymorphisms in the candidate genes. RESULTS Sequence analysis of coding regions and splice sites of the two candidate genes in 36 members including 12 hearing-impaired individuals in family Z002 failed to demonstrate any deafness-causing mutations of KCNN4 gene. There was one heterozygous mutation identified in exon10 coding sequence (942C/T) of KPTN gene, which did not result in amino acid change (P302P) as a repoerted synonymous SNP site (rs2293424). This SNP site did not cosegregate with the phenotype of family Z002. CONCLUSION Our study excluded the two candidates, KCNN4 and KPTN , as the causative genes involved in this Chinese DFNA4 pedigree.

AIM: To observe the effects of vascular endothelial growth factor(VEGF) on the proliferation and differentiation of neural stem cells(NSCs) of rats in vitro.METHODS: NSCs isolated from the hippocampal gyrus of SD rats were primary cultured and subcultured,and then divided into two groups:(1) the cells in VEGF group were treated with 150 ?g/L VEGF in the culture system,and VEGF was removed at the 7 th day;(2) control group(without VEGF treatment).The cellular morphology of two groups was observed by contrast phase microscope.Nestin and NF-200 expressing cells were detected via immunofluorescence method.The percentages of the immunostaining positive cells in each group at the 7 th day and at the 11 th day were determined.RESULTS: At the 7 th day,the percentage of nestin positive cells in VEGF group was 52.19%?7.95%,vs 29.26%?4.12% in control group(P