Background Corneal transplantation is an effective treatment to severe corneal diseases,but the shortage of cornea donor limits its application.Tissue-engineered cornea is being a new approach to corneal diseases.Objective This study was to investigate the possibility of construction of tissue-engineered corneal epithelium by culturing human amniotic mesenchymal stem cells (hAMSCs) in vitro.Methods Fresh human amniotic membranes were obtained under the approval of Ethic Committee of Affiliated First Hospital of Jinan University and informed consent of maternal women.The 6 cm×6 cm amniotic membrane tissue explant was digested using trypsin+ EDTA,and then the amniotic epithelial cells (AECs) were scraped before putting into collagenase Ⅱ digestion medium to isolate hAMSCs.hAMSCs of passage 3 were cultured to achive 80％-90％ confluence,and then the ceils were incubated on rabbit deepithelial corneal stroma at a 1 ×105/ml density.The corneal stroma was co-cuhured with hAMSCs at an air-liquid interface till 14 days.Rabbit deepithelial corneal stroma with and without hAMSCs (experimental group and control group) were fixed in 4％ para formaldehyde, and sections were prepared for histopathological examination.Immunochemistry and immunofluorescence were empoyed to detect the expressions of cytokeratin3 (CK3) and CK12 in hAMSCs.Results hAMSCs grew well and formed a stratified epidermal structure resembling native corneal epithelium on rabbit corneal stroma in cultured 14 days in the experimental group,with the oval nucleus at basement and fusiform nucleus on the surface of corneal stroma.There was no cell structure in the control group.Immunochemistry revealed brown staining for CK3, CK12 in cytoplasm of hAMSCs on the rabbits corneal stroma,and the green fluorescence for CK3 and CK12 was also seen in the hAMSCs.However,the response for CK3 and CK12 was absent in the control sections either immunochemistry or immunofluorescence test.Conclusions hAMSCs can be induced to differentiate into corneal epithelioid cells at an air-liquid interface on the rabbit corneal stroma.

Adult ; Embolism, Amniotic Fluid ; blood ; diagnosis ; surgery ; Female ; Humans ; Hysterectomy ; Pregnancy ; Uterus ; pathology ; surgery

Humans ; Immunoglobulin D ; Male ; Middle Aged ; Multiple Myeloma ; classification ; complications ; Testicular Diseases ; etiology

Child ; Child, Preschool ; Epilepsy, Generalized ; complications ; epidemiology ; genetics ; Female ; Humans ; Infant ; Male ; Pedigree ; Phenotype ; Seizures, Febrile ; complications ; epidemiology ; genetics

Objective To investigate the toxicity of levodopa and dopamine on PC12 cells and neuroprotection of several anti-Parkinson drugs i.e. amantadine,pergolide and selegiline. Methods The possible cytotoxicity of levodopa and dopamine at different dosage on rat pheochromocytoma PC12 cells and the effects of some anti-Parkinson drugs (amantadine,pergolide and selegiline) on levodopa- or dopamine-induced cytotoxicity were determined by MTT assay and flow cytometry. Results There was a concentration- and time-dependent decrease in cell viability and a concentration-dependent increase in apoptotic cells induced by levodopa and dopamine ( P

Objective To analyze the clinical curative effect of Hennipen method to treat elderly patients with anterior dislocation of shoulder joint.Methods Between February 2013 and June 2015,57 elderly patients with acute anterior dislocation of shoulder joint were colleted in our hospital.The patients aged from 65 to 81,with average age of 72.3 years.Hennipen method was used to replace the dislocation of shoulder joint.Visual analogue scale (VAS)scores and joint range of motion before and after treatment were recorded to evaluate its clinical effect.Results All cases were checked with X -ray to comfirm had been reducted,and had no complications,such as proximal humeral fractures,axillary nerve injured.1 case failed maneuver reduction,so was tracted and reseted again after anesthesia.Before treatment,the VAS scores was (5.4 ±0.9)points,which after treatment was (1.9 ±0.7)points, and the difference was significant(t =12.43,P <0.01).The range of motion of the shoulder joint was significantly improved.Conclusion Hennipen method is simple,to threat anterior dislocation of shoulder joint of elderly patients, it can effectively relieve pain,reduce complications,which is worthy of clinical promotion.

Objective To investigate the effect of chronic atrial fibrillation on free calcium concentration and expression of Ca 2+ /calmodulin dependent protein kinase Ⅱ (CaMKⅡ) in human atrial myocytes. Methods The intracellular free calcium concentration in rapidly isolated atrial myocytes and the expression of CaMKⅡ in atrial tissue of rheumatic heart disease patients with atrial fibrillation (AF) or with normal sinus rhythm were measured by laser scanning confocal microscopy and Western blotting respectively. Results The intracellular Ca 2+ concentration of patients with atrial fibrillation was significantly higher than that of patients with normal sinus rhythm [(276.38?38.12) vs (122.28?45.63) nmol/L, (P

AIM: To investigate the effect of chronic atrial fibrillation (AF) on free calcium concentration and expression of Ca 2+ /calmodulin dependent protein kinase Ⅱ (CaMKⅡ) in human atrial myocytes. METHODS: The intracellular free calcium concentration in acute isolated atrial myocytes and the expression of CaMKⅡ in atrial tissue of rheumatic heart disease patients with atrial fibrillation (AF) and with normal sinus rhythm were measured by laser scanning cofocal microscopy technique and Western blotting, respectively. RESULTS: The intracellular Ca 2+ concentration in patients with atrial fibrillation was significantly higher than that in patients with normal sinus rhythm [(276.38?38.12) nmol/L vs (122.28?45.63) nmol/L, P

Adolescent ; Branchial Region ; Fistula ; congenital ; Humans ; Male ; Neck

ObjectiveTo investigate the effect of high volume hemofiltration (HVHF) combined with mechanical ventilation (MV) on seawater respiratory distress syndrome (SW-RDS) canine models.MethodsTen nomal hybrid dogs were randomly assigned into two groups:MV group(MV group,n=5),all the animals only received MV after establishing model successfully; HVHF combined with MV group (HVHF+MV group,n=5),all were received HVHF plus MV after establishing model successfully.Both groups were observed for 4 hours.Mean arterial pressure (MAP),heart rate (HR),central venous pressure(CVP),arterial blood gas and venous plasma osmotic pressure were detected at baseline,0 min(model establishment),60 min,120 min,180 min,240min after treatment.Venous blood was collected to detect inflammatory mediators (IL-8,IL-6,TNF-α)at baseline,0 min,120 min,240 min after treatment.The lung pathology was examined at the end of the experiment.Results(1)All the animals were suvival after four hous of treatment in both groups. (2)Pattial pressure ofoxygen(PaO2) and O2 saturation(SaO2) rised after four hours of treatment in both groups(P＜0.05), and HVHF+MV group was better than MV group.After 4 hours of treatment,pH,actual bicarbonate (AB),bases excess(BE) in HVHF+MV group were significantly better than those in MV group(P＜0.05),recovering to the baseline values.(3)MAP,HR,CVP were stable during the four hours of treatment,and compared with 0 min,there was no significant differences after 4 hours of treatment in bothgroups. There were no significant differemces at the same time of treatment in both groups. (4)Plasma osmotic pressure were stable during the four hours of treatment,and compared with 0 min,there was no significant difference in MV group.But in HVHF+MV group,osmotic pressure was significantly higher after 4 hours of treatment than that at the same time in MV group(P＜0.05),and compared with 0 min and 180 min,those were higher too(P＜0.01). (5)Compared with those at the same time in MV group,plasma inflammatory mediators (IL-8,IL-6,TNF-α) were significantly decreased after 4 hours of treatment in HVHF+MV group(P＜0.01).After 4 hours of treatment IL-8,TNF-α in MV group were higher than those at the same time in 0 min (P＜0.05).(6)Compared with those in MV group,there were less infiltration of neutrophils,edema and injury of alveolar epithelium from pulmonary pathology.ConclusionsHVHF combined with MV can significantly improve hypoxemia and correct acidosis of SW-RDS model in canines.HVHF can effectively clear plasma inflammatory mediators and redundant water,improve pulmonary pathology changes.HVHF has no impact on MAP,HR and CVP of SW-RDS.