Objective:To prepare specific anti- Fusarium yolk immunoglobulin (IgY) and investigate its tolerance to temperature and pH and verify its antifungal effect against Fusarium. Methods:Eighteen 22-week-old Leghorn laying hens were selected and randomized into negative control group and experimental group according to the random number table method, with 9 hens in each group.The 2×10 7 colony forming units (CFU)/ml suspension of inactivated hyphae of Fusarium and Freund complete adjuvant was mixed in a 1∶1 ratio and emulsified.The hens in the experimental group were injected with 1 ml of the mixture to immunize and received 1 ml of Freund incomplete adjuvant as booster injection at two weeks after the initial injection.The egg yolk was collected from the 5th to 16th week after immunization.Specific anti- Fusarium IgY protein was prepared by salting out method using ammonium sulfate.The obtained protein solution was put into a freeze dryer and made into freeze-dried powder stored at 4 ℃.The hens in the negative control group were injected with 0.9% sodium chloride to prepare the non-specific antibody as the negative control.Bradford method was used to determine the concentration of specific egg IgY protein and the indirect enzyme-linked immunosorbent assay (ELISA) was employed to measure its titer.The 1×10 5 CFU/ml and 1×10 3 CFU/ml Fusarium suspension were cultured with different concentrations of IgY and phosphate buffered saline (PBS) for 4 days, and the absorbance value at 600 nm was measured.The co-incubated PBS/negative IgY with Fusarium solution was set as blank control/negative control accordingly.The concentration-killing curve of anti- Fusarium IgY against Fusarium was obtained.The specific IgY solution was diluted to 0.02 mg/ml with PBS pH 7.4, and the diluted specific IgY solution was placed into the water bath for 30 minutes at 30, 40, 50, 60, 70, 80, 90 ℃, respectively, and was cooled down to room temperature.The specific IgY solution was diluted to 0.02 mg/ml with PBS pH 1, pH 2, pH 3, pH 4, pH 5, pH 6, pH 7, pH 8, pH 9, pH 10, pH 11, pH 12, respectively, and the diluted specific IgY solution was placed at 4 ℃ for one hour.The activity of diluted specific IgY solution by different methods was measured by indirect ELISA, and the tolerance of IgY to various temperatures and pH was evaluated.Twelve 8-week-old SPF female C57BL/6 mice were selected and randomized into the PBS control group and specific IgY treatment group according to the random table method, with 6 mice in each group.The right eyes of the 12 mice were infected with Fusarium to establish mice model of fungal keratitis.One day after modeling, 200 mg/ml of anti- Fusarium IgY was dropped to the right eyes of mice in the specific IgY treatment group, and PBS was dropped to the right eyes of mice in the PBS control group.The corneas of mice in the two groups were observed under the slit lamp microscope at 1, 3 and 5 days following modeling, and the corneal ulcer was scored according to the grading scale for inflammation score.The use and care of experimental animals followed the Association for Research in Vision and Ophthalmology statement.This study protocol was approved by an Ethics Committee of The Affiliated Hospital of Qingdao University (No.QYFYWZLL26168). Results:The IgY protein concentration from the 5th to 16th week after immunization was 1.57, 2.89, 24.98, 25.09, 23.89, 25.78, 21.57, 21.37, 18.98, 15.78, 14.67, 12.67 mg/ml, respectively.The titer of IgY was increased from the 5th week, and it reached the highest titer 1∶10 000 at the 7th week, which could be maintained until the 12th week after immunization before it dropped gradually.The concentration-killing curve showed that compared with the blank control group and negative control group, Fusarium grew slowly in the specific IgY treatment group.The specific IgY with a titer greater than 1∶10 000 had thermal stability below 60 ℃.The activity of specific IgY was highest at pH 4 to 6, which could be maintained above 70% at pH 3 to 9 and was further reduced with the decrease or increase of pH.At 1, 3 and 5 days after Fusarium infection, the inflammation scores were 3.50±0.55, 7.33±0.82, 4.00±0.63 in the PBS control group, and 3.33±0.82, 4.17±0.75, 2.50±0.55 in the specific IgY treatment group.There was a statistically significant overall difference in inflammation scores at various time points between the two groups ( Fgroup=247.35, P<0.05; Ftime=23.19, P<0.05). At 3 and 5 days after Fusarium infection, there was a smaller ulcer area and decreased inflammation scores in the specific IgY treatment group compared with the PBS control group, and the differences were statistically significant (all at P<0.05). Conclusions:The high titer specific IgY can be successfully prepared by salting out method using ammonium sulfate, which is with high stability, tolerance to temperature and pH.Moreover, it can alleviate the severity of corneal ulcers and reduce inflammation scores in the mouse model of fungal keratitis.

Ophthalmology is interrelated with a variety of disciplines, and its features determine the need for interdisciplinary joint training of postgraduate students majoring in ophthalmology. The joint training can optimize the students' knowledge structures, enhance their innovation abilities and improve their overall qualities. However, there are still some factors restricting the interdisciplinary joint training in the existing education system. We propose to establish a full-time administrative management system, build a multidisciplinary communication platform, attach great importance to tutors' interdisciplinary researches, and establish dual mentor or multi-tutor training system. Moreover, a reasonable interdisciplinary evaluation system and various incentive mechanisms are advocated to improve the quality of postgraduate education in ophthalmology.

Ophthalmic pathology plays an important role in studing the occurrence,development and pathogenesis of ophthalmic diseases.Ophthalmic pathology can guide the diagnosic and treating process of primary tumor,secondary tumor and inflammatory diseases.Ophthalmic pathology,including pathological morphology,liquid biopsy,and molecular pathology,plays an important role in the connection of basic medicine and clinical medicine and guides the research of pathogenesis and promotes the development of ophthalmology.Combination of ophthalmic pathology and clinical assistant examination can provide an objective basis for precision medicine of eye disease.

Objective To study the effect of target rehabilitation diary in elderly patients with total knee arthroplasty (TKA). Methods One hundred patients who received unilateral first TKA (no patella) were numbered according to the order of admission time, according to random digit table, 50 cases in the control group and the intervention group. Conventional treatment methods were given to both two groups, the control group received routine care measures and health education, the intervention group was given target rehabilitation diary for self- management based on the conventional health education. The differences in American Knee Society Knee Score (KSS), self- management and self- efficacy were compared between the two groups. Results The KSS was (75.50 ± 6.90) points in the intervention group and (71.90 ± 7.20) points in the control group on the first week after operation, the difference was statistically significant (t=2.553, P<0.05). On the twelfth week of intervention, KSS was (85.86 ± 2.80) points in the intervention group and (80.88 ± 2.20) points in the control group, the difference was statistically significant (t=9.880, P<0.01). Self-efficacy and self-management on the twelfth week were (47.22 ± 3.34) and (54.55 ± 2.66) points in the intervention group, which were significantly higher than those in the control group (34.24 ± 4.11) and (42.54 ± 3.76) points, the difference was statistically significant (t=17.33, 18.43, P<0.01). Conclusions The target rehabilitation diary plays a good role in the self-management of elderly patients with TKA.

Objective To explore the interaction of sleep quality and depression among patients with type 2 diabetes. Methods With multistage cluster sampling, all the participants were interviewed with self-designed questionnaire, diabetes-specific quality of life scale and self-rating depression scale, and Pittsburgh sleep quality index scale. Chi-square test was used for qualitative data. Risk factors were analyzed by means of multiple linear regression or logistic regression model. The indicator of interaction was calculated according to the delta method. Results There were 944 residents involved in the final analysis, including 365 males and 579 females. The average age was (64. 0 ± 10. 2) years. Compared with patients with type 2 diabetes mellitus( T2DM) who had good sleep quality and no depression symptoms, the risk of quality of life in those with good sleep quality but depression was 2. 75 (95% CI 1. 59-4. 77); and the risk of quality of life in those with poor sleep quality and no depression was 1. 55(95%CI 1. 03-2. 33). The risk of quality of life in those with poor sleep quality and depression was 4. 97(95% CI 2. 34-9. 63). Due to poor sleep quality and depression in patients with T2DM the combined interaction index was 2. 48 (95% CI 1. 44-4. 29), the relative excess risk was 3. 42(95% CI 2. 16-4. 67), and the attributable proportion was 0. 51(95% CI 0. 32-0. 70). Conclusion A combined interaction of poor sleep quality and depression in affecting the quality of life was found in type 2 diabetic patients. When both factors existed at the same time, the interaction effect of these 2 factors was greater than the single one.

Ophthalmic microsurgery has its own characteristics: high precision , complicated operation , and long learning curve.It is important for young ophthalmologists to master the basic operation skills through standardized and systematic training during graduate studies .The teaching method , integrating theoretical teaching , animal operation and simulative ophthalmic surgery training , have important meanings in promoting the post-graduate students to master the practical and correct ophthalmic microsurgery technique . The aim of this study was to introduce the curriculum design and practice , and to evaluate the model through the examination scores .

The network teaching platform of ophthalmology of Qingdao University , as the basis construction of the national key discipline , forms a perfect and complete set of teaching system with the aid of part of course information, part of network teaching resources and part of answer and interaction. The network teaching platform focuses on the construction of network teaching resources and answer and interaction. In the college teaching of ophthalmology, through building clinical teaching resource database and discussing on network platform, we carry out interactive and discussion-based teaching, and students can prepare before class and discuss after class. The application of network teaching platform of ophthal-mology in classroom teaching and teaching feedback can improve the teaching methods , deepen the teach-ing content, implement the sharing of teaching resources, and lay a solid foundation for ophthalmology teaching reform.

Background Fungal keratitis is a common ocular surface disease and the treatment is dissatisfactory because of short of high efficiency,low toxicity and broad-spectrum medication.Penetrating keratoplasty is an effective therapy,but its clinical applications is limited due to lack of corneal donor.To seek a low-cost and effective management is of an important significance.Objective This clinical trial was to compare the clinical effectiveness between corneal ulcer debridement combined with amniotic membrane cover+drugs therapy and corneal ulcer debridement of combined drugs therapy for fungal keratitis.Methods A prospective cases-controlled study was designed.A total 210 eyes of 210 patients with fungal keratitis were included from January 2001 to January 2013 in Affiliated Hospital of Qingdao University.The patients were randomly divided into observation group (109 eyes) and control group (101 eyes).The combination therapy of corneal ulcer debridement with amniotic membrane cover and drugs was administered in the observational group,and only lesion debridement and drugs therapy was performed on the control group.The curative effect,postoperative visual acuity and therapeutic course were compared between the two groups.Results The effective rates for fungal keratitis were 100.00％,90.41％,77.78％ in the mild,moderate and severe patients,respectively,and those of the control group were 52.38％,80.00％,25.00％ respectively,showing significant differences between them (U =2.211,2.214,2.356,all at P＜0.05).The total effective rate was significantly higher in the observational group than that in the control group (91.09％ versus 75.23％),with a significant difference between them (U=3.040,P=0.002).Postoperative visual acuity was better in the observational group than that in the control group (0.44 ± 0.04 versus 0.22 ± 0.03),with a significant difference between them (t =2.225,P ＜ 0.05).The mean therapeutic course was (22.4 ± 1.8) days in the observational group,which is shorter than (33.2±2.4) days in the control group,with a significant difference between them (t =2.097,P＜0.05).Conclusions Combination therapy of lesion debridement with amniotic membrane coverage and drugs can shorten treating duration and improve vision,and it is superior to only lesion debridement and drugs therapy.

Background Studies have determined that nucleotide binding oligomerization domain 2 (NOD2) plays a key role in innate immune response.However,whether NOD2 participates in the nature defense of fungal keratitis is unclear.Objective This study was to investigate the expression and significance of NOD2 on cornea in the initial of Aspergillus fumigatus keratitis (AFK) in rats.Methods Seventy-two adult clean Wistar rats were randomized into the normal control group,only corneal epithelial scraped group and AFK model group,and the AFK models were established by incubating Aspergillus fumigatus to cornea after corneal epithelium was scraped.All the operations were performed in the right eyes of rats.Reverse transcription PCR (RT-PCR) was carried out to detect the expression of NOD2 mRNA in corneal epithelium 4,8,16,24 hours after operation.Twenty-four hours after operation,the expression of NOD2 protein in rat corneas was examined by immunochemistry and immnunofluorescence technology.Also,the rat corneas were obtained for regular histopathological examination.The use and care of the animals complied with Institutional Animal Care and Use Committee Guidebook by NIH.Results All the models were made successfully.RT-PCR revealed that a fewer NOD2 mRNA were expressed on cornea in the normal control group,but the expressing levels of NOD2 mRNA were increased in the only corneal epithelial scraped group and AFK model group.Compared with only corneal epithelial scraped group,the elevated values of NOD2 mRNA expression in the AFK model group were statistically significant at 4,8,16 and 24 hours after operation (t =-0.409,-0.439,-0.534,-0.618,all at P=0.000).The histopathological examination displayed that the cornel tissue had intact structure in the normal control group,and partly corneal epithelial deficiency,slight corneal swelling and fewer neutrophil granulocytes were seen in the only corneal epithelial scraped group.However,corneal ulcer,severe corneal edema and a lot of neutrophil granulocytes were exhibited in the AFK model group.Immunochemistry and immnunofluorescence staining evidenced that weaker expression of NOD2 was visualized in the corneal epithelial and endothelial layers,and obviously enhanced staining was seen in the AFK model group.The expressing levels (absorbancy) were 0.045 ± 0.005,0.050 ± 0.005 and 0.092 ± 0.006 in the normal control group,only corneal epithelial scraped group and AFK model group,respectively,showing a significant increase in the AFK model group compared with the only corneal epithelial scraped group (t =0.042,P =0.000).Conclusions Expression of NOD2 is upregulated in the corneas with AFK,suggesting that NOD2 participates the natural defense in the initial of fungal keratitis.NOD2 may play an important role in the process of anti-fungal innate immune response in cornea.

Objective To construct subsystem of picture archiving and communicating system (PACS) based on video database of ophthalmology and to evaluate on effectiveness of applying PACS system in postgraduate teaching.Methods PACS subsystem were constructed by collecting audiovisual materials of ophthalmology surgery videos,retinal photography,B-ultrasonography,OCT,UBM of patients.A total of 24 postgraduates were divided into control group (n =12) and experimental group (n=12) by random number table.Students in control group was taught by traditional cases,books and literature while those in experimental by network teaching from PACS subsystem.Two teaching methods were assessed subjectively and objectively.Objective indicators including ophthalmology examination results,operational skill ratings and paper reviewing results as well as subjective indicators including efficiency to accept the knowledge,satisfaction degree of teaching,influencing degree of papers were employed to assess the effectiveness of the two teaching methods.Independent samples t-test and twosample rank sum test were used for statistical analysis,P ＜ 0.05 was considered statistically significant.Results Students in experimental group done better in operation scores and in the assessment of the efficiency of knowledge accepting,satisfaction of teaching and papers effect than control group(P =0.000、0.000、0.000、0.003).The two groups had no significant difference in professional exam achievements and paper review scores (P =0.625、0.354).Conclusions PACS subsystem based on video database of ophthalmology are benefit for postgraduate teaching.