BACKGROUND:Previous studies from Shaanxi Institute of Ophthalmology have shown that ostrich cornea has the advantages to be developed into the alternatives of human corneal material.
OBJECTIVE:To determine the potential toxic effects of ostrich corneal stromal scaffold on cel s.
METHODS:Cel culture methods were used to culture L-929 cel s in the extracts of ostrich acel ular corneal
stroma which was dried and dehydrated. 3-(4,5)-Dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide assay was used to evaluate the growth and proliferation of cel s after cultured for 1, 2 and 3 days.
RESULTS AND CONCLUSION:After the cel s were cultured in the extracts of ostrich acel ular corneal stroma subjected to dryness and dehydration for 1, 3 and 5 days, and the toxicity level of cultured cel s was graded as level 1. The cytotoxicity test was conducted according to the“National Standard of the People's Republic of China GB/T16886.5-2003”. After cultured in the extracts of ostrich acel ular corneal stroma, a smal number of cel s were round in shape and loosely adherent without intracytoplasmic granules, and cel lysis could be observed
occasional y. The results of 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide assay showed that
the ostrich acel ular corneal stromal scaffold which was dried and dehydrated had level 1 of cytotoxicity and could be considered as a qualified material.

Combined with the clinical practice of Xi'an Eye Bank for over four decades,this article focuses on the ethical issues related to the practice of eye bank,such as voluntary cornea donation free of charge,the relationship between donors and recipients,the choice of operation indication and application of heterogeneous cornea,in order to promote the all-round development of eye bank system.

Background Ostrich acellular corneal stroma possesses a similar constitution to human corneal stroma,so it is expected to become one of ideal biological corneal carriers.Objective This study was to investigate the immunogenicity of acellular stroma carrier of ostrich cornea and offer the information for the development of industrialization and clinical use of acellular stroma carrier of ostrich cornea.Methods Twenty fresh ostrich eyeballs and 20 porcine eyeballs were collected.Acellular corneal stroma carriers of ostriches and pigs were prepared using low temperature freezing joint enzyme digestion method and desiccant dehydration method and sterilized by cobalt-60 irradiation.The corneal stroma carriers were preserved using drying and dehydration method.Forty-five male BALB/c mice were randomly divided into the sham operation group,ostrich acellular corneal stroma group and porcine acellular corneal stroma group.Acellular corneal stroma carriers of ostriches and pigs(wet weight after rewatering was 10 mg/piece) were subcutaneously implanted to the back of BALB/c mice,respectively.Wound healing and inflammatory response on the operative site were observed,and phenotype and activating rate of CD4+,CD8+ and CD25+in peripheral blood of mice were dynamically detected 7,14 and 28 days after ectopically implantation of heterogeneous corneal stroma by immunofluorescence labeling and flow cytometry analysis.Results No swelling and exudation were seen in the skin of operative site of the mice with a good healing of wound after surgery.There were no significant differences in the activating rates of CD4+,CD8+ and CD25+ cells in the peripheral blood of mice among the sham operation group,the porcine acellular corneal stroma group and ostrich acellular corneal stroma group in the three time points after surgery(CD4+:F=0.74,P=0.50;F=0.39,P=0.05;F=3.46,P=0.58.CD8+:F=1.75,P=0.21 ;F=1.14,P=0.35;F=0.78,P=0.48.CD25+:F=0.52,P=0.61 ;F=3.53,P=0.62;F=2.42,P=0.13).Conclusions The ostrich acellular heterogeneous corneal stroma carrier possesses low immunogenicity.It is inferred that ostrich acellular corneal stroma carrier can be used in heterogeneous corneal transplantation.

Objective To select the largest non-toxic leaching solution concentration through the experimental observation of the cytotoxicity of the ostrich acellular corneal stromal leaching solution to the Chinese hamster lung fibroblasts cells(CHL) for the further chromosome distortion experiment.Methods The leaching solution made from the ostrich acellular corneal stromal material was diluted with concentrate of 1 ∶ 2,1 ∶ 4 and the original concentration were used to culture with the CHL cells,the negative and positive control were also set up at the same time,to evaluate the impact on cell growth after 24 hour by MTT colorimetric method.Results The leaching solution diluted with 1∶4 was non-toxic,and could promote the growth of the cells.Conclusion Combined with the results of classification and cell morphological features,this cytotoxicity test can be used to screen the best benchmark non-toxic concentrations for the chromosome aberration test of the CHL cells.

Objective To investigate the difference of CCAAT/enhancer-binding protein α (C/EBP-α) gene induced apoptosis between hepatocytes and hepatic stellate cells (HSC) in mice with liver fibrosis.Methods Sixty BALB/c mice were evenly divided into normal group,model group,treatment group,blank control group and negative control group,12 mice in each group.Except the mice of normal control group,the mice of other groups were treated with intraperitoneal injection of CCl4 to establish liver fibrosis mice model.Mice of treatment group,blank control group and negative control group were administrated with C/EBP-α carried adenovirus (Ad-C/EBP-α),phosphate buffered solution and empty vector of adenovirus (Ad-EGFP) respectively through tail vein for the first week.The expression of C/EBP-α and α-smooth muscle actin (α-SMA) was detected by immunohistochemistry method.Sinusoidal endothelial structure of peri-portal regions and far from portal regions was observed by transmission electron microscope (TEM).Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was applied to detect apoptosis of cells in liver tissue.The degree of liver fibrosis in mice was determined with sirius red staining and hydroxyproline content measurement.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between two groups.Results C/EBP-α was expressed in nucleus of hepatocyte in normal control group mice.The expression decreased in model group,blank control group and negative control group.However,the expression of C/EBP-α of treatment group increased,and mainly expressed in cells located in perisinusoidal and perivascular.Hepatic sinusoids was distorted,blood vessel wall thickened.Hepatocyte degeneration and lots of lipid droplets was found in model group,blank control group and negative control group.The thicken degree of endothelial layer of blood vessel of treatment group was lower than that of model group.The percentage of sirius red positive cells of normal group,model group,treatment group,blank control group and negative control group was (0.10±0.03)％,(5.81±0.32)％,(2.32±0.45)％,(6.34± 0.81)％ and (6.10± 0.92)％,respectively; content of hydroxyproline was (0.07±0.00) μg/mg,(0.69 ± 0.10) μg/mg,(0.19±0.06) μg/mg,(0.56±0.03) μg/mg and (0.64±0.08) μg/mg,respectively; the percentage of α-SMA positive cells was (0.50 ±0.03)％,(5.30 ± 0.52)％,(2.15 ± 0.29)％,(5.53 ± 0.43) ％ and (5.42 ± 0.25) ％,respectively; the number of TUNEL positive cells was (0.25 ± 0.08),(0.15±0.02),(7.10±1.53),(0.13±0.03) and (0.18±0.07),respectively.The differences between the groups were statistically significant (F=113.74,148.29,292.43 and 140.25,all P＜0.05).The difference between normal group and model group,between model group and treatment group,between treatment group and blank control group,between treatment group and negative control group were statistically significant (tarirus positive cell =-52.54,-16.20,-10.60 and-7.99,thydroxyproline content =-168.00,11.53,11.07 and 12.54,ta SMA pusitive cells-24.77,-13.82,15.94 and 18.37,tTUNEL positive cells =3.26,-11.91,-11.95 and-11.88,all P＜ 0.05),there was no statistically significant difference between model group and blank control group,between model group and negative control group (both P＞0.05).TUNEL positive cells mainly located in perisinusoidal and perivascular of liver in mice,which was consistent with the distribution of α-SMA-positive cells.Conclusion C/EBP-α could effectively relieve CCl4 induced liver fibrosis in mice mainly through inducing HSC apoptosis,however no apoptosis effect on hepatocytes.

OBJECTIVE:To observe the analgesia effects of transdermal fentanyl in the treatment of pharynx oralis pain caused by radiotherapy.METHODS:30patients with pate squamous carcinoma and serious membrana mucosa ulcerate and pain after a whole range radiotherapy,who were treated with3to5patches2.5mg(25?g/h)of transdermal fentanyl every9～15days if their VAS scores were found to be more than6scores.The analgesia effect,life quality and the adverse reaction were evaluated respectively with VAS,Digit-scale and east American carcinoma cooperation group toxicity criteria.RESULTS:The average VAS scores has been lowered from7.86?1.18before the treatment to2.24?1.31on10th day of the treatment(P

Objeetive To study the differences of the biological characteristics and immune regulation function of adipose mesenchymal stem cells (AMSCs)from psoriasis patients and healthy people.Methods AMSCs were isolated and cultured from human psoriatic and healthy adipose tissue,the phenotypes and cell cycle of AMSCs taken from three generation were detected by flow eytometry.Alkaline phosphate enzyme staining and oil red o staining were used respectively to identify their adipogenic and osteogenic capacity.Next,the levels of inflammation antimicrobial proinflammatory factor were detected by PCR and ELISA.Then gene expression profile of AMSCs were screen by gene expression profile chip,as so to bolting the the gene array related with immunology gene.Results There was no significant change in cell morphology,and cell surface markers were expressed high for CD29,CD44,CD73,while lower for CD31,CD45 and HLA-DR.AMSCs of psoriasis patients and healthy people both had the ability of adipogenic and osteogenic differentiation.But the cell cycle showed the third generation AMSCs proliferation rates were slower than that of normal control,as compared with healthy controls,adipogenic differentiation ability was stronger.What'more,the level of inflammatory cytokines in psoriasis group was lower than that in controls such asIL-10,IDO,TGF-β,on the contrary the levels of proinflammatory factor in psoriasis group were higher than that in controls,such as TNF-α,IFN-γ.In addition,gene chip results suggested that psoriasis group AMSCs had obvious expression differences on JAK-STAT pathway with healthy controls.Conclusions Compared with the control,there are significant differences in patients AMSCs proliferation and adipogenic differentiation ability,immune inflammation suppression control ability is weaken,this phenomenon may be associated with JAK-STAT immune pathways related to downgrade.

Objective To evaluate the clinical value of ultrasonography in the diagnosis of hermaphroditism.Methods Ultrasonographic appearances of 45 cases with hermaphroditism were reviewed retrospectively,and the results were compared with clinical data.Results All cases were confirmed pathologically.5 cases showed true hermaphroditism (11.1％),14 cases showed male pseudohermaphroditism (31.1 ％),24 cases showed female pseudohermaphroditism (53.3 ％),2 cases showed gonadal dysgenesis(4.4％).Compared with pathological results in the ultrasound examination,4 cases showed true hermaphroditism,the coincidence rate was 80％.11 cases showed male pseudohermaphroditism,the coincidence rate was 78.6％.20 cases showed female pseudohermaphroditism,the coincidence rate was 83.3％.1 cases showed gonadal dysgenesis,the coincidence rate was 50.0％.Conclusions Ultrasonography can not only evaluate internal reproductive organs in the patients,but also estimate the site,size,morphology and structure,which provides important imaging evidence for the clinical diagnosis and treatment.

OBJECTIVE:To establish a new semi-automatic drug dispensing mode,with proper cost,which falls in between full-automatic drug dispensing mode and manual drug dispensing mode,good work efficiency,standard and simple operation meth-od and meets national laws and regulations. METHODS:A semi-automatic drug dispensing system was designed,in which the in-formation in the drug dispensing sheet could be automatically printed on the drug bag,and automatic bagging,packaging and deliv-ery of drugs were realized. Such drug dispensing system included hardware(mechanical structures such as drug turntable and drug funnel,transmission device,etc.)and software control systems(the program of interface with hospital information system,micro control unit software,computer software,etc.). Through commissioning,formal operation and statistics,based on 18 oral drug dis-pensing sheets with the same contents,calculated the time of drug dispensing and the number of drug dispensing errors by 3 phar-macists respectively in manual drug bag dispensing mode and semi-automatic drug dispensing mode,to evaluate the effect of the semi-automatic drug dispensing system. RESULTS:From commissioning in May 2012 to formal operation in September 2012,the system operated normally and utility model patents were obtained. In the two modes,the total time of drug dispensing was 481 and 397 min (t=6.82,P<0.001),the numbers of drug dispensing errors were 25 and 7 (χ2=9.353 8,P=0.002 2),respectively. There was statistical significance. CONCLUSIONS:The semi-automatic drug dispensing system has higher efficiency and less num-ber of drug dispensing errors compared with manual drug bag dispensing mode and lower cost compared to full-automatic drug dis-pensing system. It deserves promotion.

Objective To culture and identify corneal fibroblasts from human keratoconus patients (HKCs).Methods HKCs and corneal fibroblasts from human healthy controls (HCFs) were cultured by tissue block adherence method.Cellular morphology and ultrastructure were observed by inverted phase contrast microscope and electron microscopy respectively.Cell viability was detected by cell counting kit-8 (CCK-8) assay.α-Smooth muscle actin (αt-SMA),collagen type 1 alpha 1 (COL1A1) and collagen type 3 alpha 1 (COL3A1) protein expression levels were detected by Western blot.This study protocol was approved by Ethic Committee of Xi'an No.1 Hospital (No.1504).Results Compared with HCFs,HKCs showed several distinguishing properties.First of all,its growth speed was faster,with collagen fibers decreased and attenuated.At the same time,mitochondrion swelled and mitochondrial cristae disappeared.Additionally,Golgi apparatus presented significant expansion and endoplasmic reticulum displayed severe swelling.There were statistically significant differences in A values between the two kinds of corneal fibroblasts at different time points after culture (Fgroup =5 023.13,P＜0.01;Ftime =38 518.16,P＜0.01),the A value of HKCs was significantly higher than that of HCFs at the same time point,and the difference was statistically significant (all at P＜0.01).The relative expression of α-SMA,COL3A1 and COL1 A1 was 120.00±5.77,158.33 ±4.41 and 88.33± 1.67,respectively in HKCs,the relative expression of α-SMA,COL3A1 and COL1 A1 was 100.00±0.00,100.00±0.00 and 100.00±0.00,respectively in HCFs,the relative expressions of α-SMA and COL3A1 were significantly increased in HKCs than those in HCFs,the relative expression of COL1 A1 was significantly decreased in HKCs than that in HCFs,with significant differences between them (t =-3.46,P ＜ 0.05;t =-13.23,P ＜ 0.01;t =7.00,P＜0.05).Conclusions HKCs are cultured and identied,which is suitable for establishing in vitro cell model of keratoconus.