Objective To explore the plasticity of transdifferentiation of epidermal stem cells (ESCs) into corneal epithelial cells. Methods Epidermal stem cells enriched by adhering to type Ⅳ collage were induced in vitro by coculture with corneal stromal fibroblasts. The method of HCC were used to show the expression of K12 which is expressed specially in corneal epithelial cells.Results ESCs with greater proliferation potential strongly expressed K19 and ?_ 1 - integrin. After two-week induction, the K12 positive cells could be detected. Conclusion The data suggested that the epidermal stem cells have the potential to transdifferentiate into corneal epithelial cells in vitro.

Objective To isolate and culture the hair follicle Bulge cells of rats and study their morphological, immunological characteristics. Methods Hair follicle Bulge obtained by micromanipulation and enzyme digestion was cultured in intro. The morphological features and numbers of Bulge cells were identified and counted with light microscopy. Immunocytochemical staining was used to detect expressing of K19 and ?_ 1integrin in cultured cells. Results Bulge cells after enzyme digestion pasted rapidly. Cells were small and round, paving stone-shape, and had a large ratio of nuclear to cytoplasm. The characteristics suggested a primitive morphologic feature. Cells growth well and could maintain low differentiation and higher reproductive activity, they co-expressing K19 and ?_ 1integrin, immunofluorescence staining showed the cells expressed ?6-integrin strongly, weakly or no expressing CD71. Conclusion This culture system can amplify a lot of pure hair follicle Bulge cells in short time. We successfully isolate and culture the hair follicle Bulge cells of rats in intro and can keeping its property for a long time.

Objective To explore the roles of urokinase type PA (uPA) and uPA receptor (uPA R) in the course of repair after human embryo corneal alkali burn in vitro . Methods After the alkali burn model in vitro was established successfully, some techniques such as the observation of cell culture and morphology, ICC and image analysis were used. Results No significant difference was found between the cells from corneal limbus and central cornea and almost all of them expressed AE1/AE3. The expressions of uPA and uPA R increased to the maximum at about 24 h after alkali burn. uPA expression distributed mainly in the cytoplasm but uPA R expression distributed mainly in the cell membrane. Conclusion The corneal epithelial cells of comparatively high purity can be acquired by tissue culture. There probably exists difference in development and vitality between embryo and adult cornea. There might be commonness of the roles of uPA and uPA R in epithelial tissue.

OBJECTIVE:To provide reference for rational utilization of antibiotics for special use. METHODS:Medical orders of antibiotics for special use in clinical departments of our hospital during 2013 to 2015 were selected from hospital information sys-tem. The consumption sum and its ratio,DDDs,DDC,DUI and the utilization of antibiotics for special use in clinical departments were calculated and analyzed. RESULTS:The consumption sum and its ratio of antibiotics for special use in our hospital during 2013 to 2015 both reduced year by year,decreasing from 21 872 200 yuan(48.00%)to 20 877 700 yuan(39.41%). The consump-tion sum ratio of carbapenems,cephalosporins and anti-deep fungal drugs showed descending tendency. The consumption sum ratio of anti-MRSA antibiotics changed slightly. DDDs of Meropenem for injection always took up the first place in recent 3 years,but the values were decreasing. DUI of Imipenem and cilastain sodium for injection was far less than 1,while those of Panipenem and betamipron for injection,Cefepime sodium for injection and Caspofungin acetate for injection(70 mg/injection)were far more than 1. The types of drug with DUI ranged 0.9-1.1 increased year by year,increasing from 3 types in 2013 (18.75%) to 10 types in 2015 (62.50%). Within 3 years, the utilization ratio of antibiotics for special use always took up the first place in ICU (86.64%-87.78%). CONCLUSIONS:The consumption sum and its ratio of antibiotics for special use in our hospital during 2013 to 2015 decreased year by year,and the utilization of antibiotics for special use become increasingly rational. But,there still are some problems,such as inadequate dose of Imipenem and cilastain sodium for injection,overdose of Panipenem and betamipron for injection,Cefepime sodium for injection and Caspofungin acetate for injection(70 mg/injection).

Objective To investigate the feasibility of the transdifferentiation of hair follicle bulge cells into corneal epithelial cells in vitro. Methods The hair follicle bulge cells from 7- to 8-day SD rats were isolated and cultured in vitro, then co-cultured with rabbit corneal limbal stroma in transwell-cultured system. The differentiation and development of hair follicle bulge cells were observed, and immunocytochemical staining were used to detect expression of K19 and K12 in hair follicle bulge cells. Results The cultured bulge cells possesed high proliferation and low differentiation, co-expressed K19 and ?_ 1 integrin, but part of them expressed K12 after 2-week co-culture with rabbit corneal limbal stroma in transwell-cultured system. Conclusion Rat hair follicle bulge cells could transdifferentiate into corneal epithelial cells induced by corneal limbal stroma in vitro.

Objective To investigate the diagnostic value of acridine orange fluorescene(AO-F) in bladder cancers. Methods One thousand and sixteen bladder cancer patients were reviewed retro-spectively. The positive-rates of AO-F in different stages, grades, size, quantity, position of tumors, hematuria and treatment ways were evaluated. Results The total positive rate of AO-F was 78.05 % (793/1016). The positive-rate was 74.69% (611/818) in superficial stage and 91.92% (182/198) in invasive bladder cancer, 67.24% (351/522) in grade Ⅰ and Ⅱ , 90. 37% (413/457) in grade Ⅲ. The percentage of positive AO-F was 80.30% (750/934) in patients with hematuria, 52.44% (43/82) in patients without hematuria. The percentage was 79.87% (710/889) when the tumor size was more than 2 cm, 65.35% (83/127) when size less than 2 cm. 83.07% (363/437) sample was positive in multiple tumors, 74.27% (430/579) in single tumor. The percentage was 77.21% (105/136) in tumors involving trigone or neck of bladder, 78.07% (687/880) in tumors without involving these re-gions. There was 69.68% (393/564) in treatment with TURBt, 87.87% (268/305) in partial resec-tion, 91.74% (100/109) in total resection. A good association was observed between stage, grade, hematuria appearance, tumor size, quantity of carcinoma, treatment way and AO-F positive-rate, and a linear correlation was present between grade, stage and positive cytology. There was no significant association between position of the tumor and AO-F positive-rate. Conclusions The function of AO-F is significant in diagnosis of bladder cancer.

Objective To observe the growth characteristic of bulge-originated cells from human hair follicles in vitro. Methods The bulges which were isolated from human hair follicles by dispase and microdissection were cultured. The morphological and biological characteristic of the cultured bulge cells were observed by light microscopy and immunocytochemistry (ICC). Results Proliferated cells could be observed in the second day after being inoculated. The number of these cells, with greater proliferation potential, reached the peak at the sixth day and maintain several days. In addition, the mitotic figures appeared and the cells behave the similar morphologic features, meanwhile the cells strongly expressed K19 and showed a descensive tendency in long-time growth.Conclusion The cultured bulge cells kept the primitive characteristic, which suggested that the putative follicle stem cells resided in the bulge area.;

Objective To study the function of cyclooxygenase 2(COX 2) and its specific inhibitor NS 398 on the cell growth and invasion ability of urothelial carcinoma cell line EJ. Methods The cox 2 cDNA was transfected into the urothelial carcinoma cell line EJ and a cell line EJ COX 2 which highly expressed cox 2 gene permanently was gained. The cell growth rate before and after transfection was observed. Then at various concentrations of NS 398, the invasion ability was detected by Boyden Chamber and expression levels of uPA by RT PCR and Western blot. Results The EJ COX 2 cell line grew more rapidly and had a stronger invasion ability than EJ and its uPA expression increased significantly. NS 398 could dose dependently inhibit the expressions of COX 2 and uPA and the invasiveness of EJ COX 2 cell. Conclusion COX 2 can stimulate the growth of urothelial cell line EJ and promote its invasion ability by stimulating the expression of uPA.

Objective To isolate and culture human adipose-derived stem cells (hADSCs),investigate the influence of hADSCs on the cellular morphology,survival rate,and function of human islet cells under the in vitro non-contact co-culture conditions,and explore its mechanism.Method hADSCs were isolated by collagenase digestion method,then cultured,and identified by morphology,immunofluorescence and multi-directional differentiation.Adult islet cells were separated and purified by Liberase enzyme and Ficoll 400,then divided into co-culture group and individual group.The cellular growth morphology of islet cells was observed by inverted phase contrast microscope.The survival rate of islet cells,insulin secretory volume,insulin stimulation index and concentration of growth factor in the supernatant were compared between the two groups.Result hADSCs of the third generation showed uniform long spindle fibrocyte-like morphology,and had multi-directional differentiational potentials of osteogenesis and adipogenesis.Immunofluorescence test of surface antigens on hADSCs revealed CD44 + and CD49d +,CD31-,CD34-and CD106-.After 14-day culture,the islet cellular morphology in co-culture group was more intact than that in individual group.The survival rate of islet cells in co-culture group was (82.83 + 2.32) ％,and that in individual group was (53.00 + 2.82) ％ (P＜0.01).Insulin secretory volumes were (23.66 + 2.11) and (7.82 +1.09) mU/L respectively in co-culture group and individual group under high glucose concentration,and 13.22 + 0.77 and 6.40 + 0.44 mU/L respectively under low glucose concentration (P＜0.01 for all).Insulin stimulation index was decreased from 1.67 + 0.10 (at 3rd day) to 1.77 + 0.13 (at 14th day) in co-culture group,and from (1.67 + 0.10) (at 3rd day) to (1.77 + 0.13) (at 14th day) in individual group (P＜0.01).After 14-day culture,the concentrations of HGF,TGF-β,VEGF and bFGF in the supernatant were higher in co-culture group than in individual group (P＜0.01).Conclusion hADSCs were isolated and cultured successfully from adult adipose tissue.They could increase the survival rate and improve the function of islet cells when co-culture with the adult islet cells in vitro through secreting HGF,TGF-β,VEGF and b-FGF.

ABSTRACT:Objective For preparation of vascularized islets , to isolate and culture human adipose derived stem cells , investigate the role of adipose derived stem cells (ADSCs ) in promoting the proliferation and vascularization of human umbilical vein endothelial cells (HUVECs ) co‐cultured in vitro , and explore its mechanism .Methods ADSCs and HUVECs were isolated by collagenase digestion method ,then cultured ,and identified by morphology ,immunofluorescence or multi‐directional differentiation .The co‐culture system of ADSCs and HUVECs was established , HUVECs cultured alone were set up for control group . The proliferation , vascularization and concentration of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b‐FGF)in the supernatant were compared between the two groups .Results The third generational ADSCs had uniform long spindle fiberous morphology and multi‐directional differentiational function . Immunofluorescence test of surface antigens on ADSCs revealed CD44/CD49d (+ ) ,CD31/CD34 (-) ,on HUVECs CD31/vWF (+ ) . High vascular density was found when co‐cultured in Matrigel of ADSCs and HUVECs than alone of HUVECs .Growth curve shown at days 3 , 4 and 5 of the logarithmic phase , HUVECs count in co‐culture group of ADSCs and HUVECs was (4 .52 ± 0 .31) × 104 ,(7 .18 ± 0 .45) × 104 ,and (8 .23 ± 0 .36) × 104 under indirect co‐culture condition , while that in individual HUVECs group was (2 .71 ± 0 .25) × 104 ,(4 .87 ± 0 .26) × 104 ,and (6 .86 ± 0 .33) × 104 ( P<0 .01) .Population doubling time of HUVECs was shorter in co‐culture group than in individual group .Also ,the OD value of HUVECs was higher in co‐culture group than in individual group when cultured at days 1 ,3 ,5 and 7 ( P<0 .01) .When cultured at days 3 ,7 and 13 ,the concentration of VEGF and b‐FGF in the supernatant was higher in co‐culture group than in individual group ( P< 0 .01 ) . Conclusion ADSCs can promote the proliferation and vascularization of HUVECs in vitro co‐culture conditions by secreting or increasing the HUVECs secretion of VEGF and b‐FGF .