Background Diclofenac sodium eye drops,pranoprofen eye drops and bromfenac sodium hydrate eye drops are three clinical commonly used nonsteroidal anti-inflammatory drugs(NSAIDs).The variation of cytoxicity among these drugs and whether the cytoxicity is related to the supplements are also unknown.Objective This study was to compare the cytotoxicity of three non-steroidal anti-inflammatory eye drops and their active components with cultured human corneal epithelial cells (HCECs) in vitro,and to discuss toxic origins of these drugs.Methods HCECs were cultured in different drugs with the final concentration of 0.10％,0.05％,0.02％ and 0.01％.Cell proliferation was evaluated by MTT assay.Then,0.002％ eye drops (1∶50) was added,and the migration and damage of the cells were deceted by transwell migration assay and lactate dehydrogenase (LDH) assay.Results The cytotoxicity of three nonsteroidal anti-inflammatory eye drops on HCECs was concentration-dependent (all at P=0.00).Diclofenac sodium eye drops showed the most dominant effects on the proliferation,migration and damage of HCECs among the three eye drops,while bromfenac sodium eye drops showed the least effect on the cell damage (proliferation:Fdrug =20.25,P=0.00;migration:F =103.43,P =0.00;damage:Fdrug =164.16,P =0.00).Compared with the eye drops,their active components showed less cytoxicity.Pranoprofen appeared the least effects on the proliferation,migration and damage of HCECs (proliferation:Fdrug =332.27,P =0.00;migration:F =332.27,P =0.00;damage:Fdrug=154.83,P=0.00).Conclusions The cytotoxicity ofdiclofenac sodium eye drops is more obvious than that of pranoprofen eye drops or bromfenac sodium hydrate eye drops.The cytotoxicity of the three eye drops originates from their supplements or the interaction between the supplements and active components.

Background Limbal stem cells (LSCs) play an important role on the stability of corneal epithelium and corneal transparency.Rho-associated coiled-coil containing protein kinase (ROCK) inhibitor can promote cell proliferation and reduce apoptosis,such as human embryonic stem cells and karatin epithelial cells.Objective This study was to investigate the improving effect of Y-27632,a ROCK inhibitor,on the activity of rabbit LSCs in corneal preservation medium.Methods Corneal preservation solution was prepared by adding 12.5％ chondroitin sulfate,10.0％ low molecular dextran,20.0 mg/L dexamethasone,100 mg/L tobramycin sulfate,9.5 g/L Hepes and 0.375 mg/L L-glutamine in MEM.The corneas of New Zealand white rabbits were collected and preserved in the corneal preservation solution with or without Y-27632 for 4,7,14 days,and the density and morphology of corneal endothelial cells were examined by using 0.25％ trypan blue staining and 0.2％ alizarin red staining.Isolated corneal epithelial cells were seeded on 3T3 feeder layer and cultured for 7-10 days until colonies formation.Colony shape of LSCs was observed under the light microscope,and colony-formation efficiency was analyzed after Giemsa staining by Image J software.Results The morphology and density of corneal endothelial cells were normal in the corneal preservation solution with and without Y-27632 for 4 days.In the seventh day after preservation,the cells remained the regular hexagon in shape in the preservation solution with Y-27632,however,the cellular membrane was slightly shrinking with the positive staining for alizarin red in the preservation solution without Y-27632.The density of corneal endothelial cells in the corneal preservation solution without Y-27632 was (2 262-± 75) cells /mm2,while in the preservation solution with Y-27632 was (2 425 ±95) cells/mm2(P＜0.001).The cloning spheres of LSCs were similar in preservation solution both with and without Y-27632 in the freshly isolated cornea or preserved corneas and exhibited more cells inside.But in 7 days and 14 days after preservation,the cloning spheres were much smaller in the preservation solution without Y-27632 group than those in the preservation with Y-27632 group.No significant differences were found in the cloning-formation rate and survival rate of corneal epithelial cells in corneas freshly isolated or preserved for 4 days in both groups (all at P＞0.05).In 7 days and 14 days after preservation,the active rates of corneal epitheli.al cells were (73.00±2.12)％ and (56.00±0.71)％ in the preservation solution with Y27632,which were significantly higher than (66.00 ± 4.00) ％ and (49.00 ± 0.71) ％ in the preservation solution without Y-27632,showing statistically significant differences between them (t =3.098,P =0.018;t =9.798,P =0.000).In addition,the cloning-formation rates of LSCs were (11.05±0.21)％ and (3.10±1.97)％ in the preservation solution with Y-27632 in 7 days and 14 days after preservation,revealing significantly elevation in comparison with (2.05 ± 1.20) ％ and (0.40 ±0.14) ％ in the preservation solution without Y-27632 (t =18.107,P =0.000;t=3.184,P=0.017).Conclusions Y-27632 promotes the vitality and cloning-formation ability of LSCs in corneal preservation medium,suggesting its potential use during storage of cornea.

Objective To study the over-expression and clinical implications of the oncogene MDM2 in acute leukemia (AL). Methods The expression of MDM2 gene in 100 patients with newly diagnosed and relapse or refractory AL and 20 healthy as control was measured by relative quantitative reverse transcriptase polymerase chain reaction (RT-PCR),then the results was measured by χ2-test,t-test and one-way ANOVA to compare expession positive rate and intensity of MDM2. Results Among 100 patients,fifty-eight had the high expression of MDM2 gene (58 %). The expression level of MDM2 gene in patients was higher than that of health controls(P ＜0.05). The expression positive rate of MDM2 is higher in poor outcome group (67.9 %,19/28)than that in general outcome group (33.9%,19/56) (P＜0.05). Conclusion Our results suggest that the expression of MDM2 gene plays an important role in the pathogenesis and poor outcome of AL.

Objective To evaluate the application effect of"2+2"medicine-education cooperation talents training mode. Methods The nursing undergraduates of 2013 grade were divided into the experimental and control group. The"2+2"medicine-education cooperation talents training mode was adopted in the former and traditional training model was used in the latter. The final examination scores of the fifth and sixth semester, scores of graduation examination, scores of the Nurse Competence Scale were compared between the two groups. Results There was no difference of scores of theory examination between the two group except the Nursing Professional English whose scores were higher in the control group. The practical skills exam scores of Medical Nursing, Surgical Nursing, Paediatric Nursing, Nursing of Gynecology and Obstetrics, Emergency Nursing, Rehabilitation Nursing, Community Nursing in the experimental group were 89.32 ± 6.02, 89.46 ± 5.29, 88.06 ± 6.34, 89.32 ± 5.58, 83.99 ± 6.44, 82.58 ± 5.78, 83.56±6.12, the control group were 87.72±5.90, 87.85±6.32, 85.59±5.79, 87.45±5.65, 81.82±6.06, 80.34± 5.89, 81.28±5.42, and the differences were statistically significant (t=2.052-3.261, P<0.05). The total and practical skills scores of graduation examination in the experimental group were 82.86±4.92, 85.60±4.54, which were 81.07 ± 5.52, 84.07 ± 4.59 in the control group. There were significant differences between the two groups (t=2.060, 2.011, P<0.05). The total score of Nurse Competence Scale and scores of the four dimensions of Teaching-coaching, Managing situations, Ensuring quality, Work role were 74.95 ± 3.40, 76.38 ± 3.98, 75.49 ± 3.23, 75.42 ± 2.72, 77.49 ± 2.76 in the experimental group, the control group were 73.63 ± 4.39, 72.90 ± 4.23, 74.12 ± 4.19, 74.32 ± 3.36, 76.46 ± 3.24, respectively. The differences were statistically significant (t=2.021- 2.492, P<0.05). Conclusions The"2 + 2"medicine- education cooperation talents training mode is proved to be an effective way to train nursing applied talents to meet regional demands. It could improve the hospital′s synthetic strength including health care quality and teaching ability, promote the development and construction of the nursing specialty and train the double-certificate teachers.

Vincristine, a widely used chemotherapeutic agent for treating different cancer, often induces severe peripheral neuropathic pain. A common symptom of vincristine-induced peripheral neuropathic pain is mechanical allodynia and hyperalgesia. However, mechanisms underlying vincristine-induced mechanical allodynia and hyperalgesia are not well understood. In the present study, we show with behavioral assessment in rats that vincristine induces mechanical allodynia and hyperalgesia in a PIEZO2 channel-dependent manner since gene knockdown or pharmacological inhibition of PIEZO2 channels alleviates vincristine-induced mechanical hypersensitivity. Electrophysiological results show that vincristine potentiates PIEZO2 rapidly adapting (RA) mechanically-activated (MA) currents in rat dorsal root ganglion (DRG) neurons. We have found that vincristine-induced potentiation of PIEZO2 MA currents is due to the enhancement of static plasma membrane tension (SPMT) of these cells following vincristine treatment. Reducing SPMT of DRG neurons by cytochalasin D (CD), a disruptor of the actin filament, abolishes vincristine-induced potentiation of PIEZO2 MA currents, and suppresses vincristine-induced mechanical hypersensitivity in rats. Collectively, enhancing SPMT and subsequently potentiating PIEZO2 MA currents in primary afferent neurons may be an underlying mechanism responsible for vincristine-induced mechanical allodynia and hyperalgesia in rats. Targeting to inhibit PIEZO2 channels may be an effective analgesic method to attenuate vincristine-induced mechanical hypersensitivity.