Ocular surface is a special interface between the inner ocular tissue and the external environment.It provides effective physical and biological barriers to prevent environmentally harmful substances and pathogenic microbes invading into the eye.This protection is afforded by unique local anatomy and cellular components, especially the resident immune cells such as innate lymphoid cells (ILCs), macrophages, mast cells, Langerhans cells(LCs) and γδ T cells.Recent studies reinforce the importance of LCs and mast cells as inducers of immune tolerance.Most adult tissue macrophage populations are seeded before birth and maintained in the steady state independently of monocytes from blood.However, under inflammatory condition, some transient monocytes enter into tissue and become macrophage compartments.Monocytes are subdivided into two main phenotypically and functionally distinct subsets.The first main subset is dedicated to the surveillance of endothelial integrity.The second main subset compasses classical monocyte functions such as rapid migration to the sites of injury and the replenishment of peripheral dendritic cells and macrophage compartments.In addition, newly identified ILCs are founded in the ocular surface tissues.These cell groups defens the different stimulus from external environment by producing different cytokines and growth factors.Accumulating data highlight some key roles of tissue-resident immune cells in ocular surface homeostasis and pathology.

Objective To observe the dynamic changes of adrenomedulin(ADM) and calcitonin-gene-relat-ed peptide(CGRP) at different time before,during and after laparoscopic operation for treating CO2 pneumoperitone-urn. Methods 47 cases undergoing laparoscopic operation in our hospital were selected. Plasma ADM and CGRP levels were determined by RIA before anesthesia(T0),10 min before (T1) and 10 min (T2) ,30 min (T3),60 min (T4),and 30 min (TS) after CO2 pneumoperitoneum was treated. Results There was statistical significance in ADM and CGRP level at the different time points before and after CO2 pneumoperitoneum (F=9.686 and 49.804, P<0.01). There was no significance in ADM and CGRP level at TO and T1 (P>0.05);ADM level would decrease at T2 (P<0.05) but CGRP level decrease slowly with no remarkable change as compared with that before CO2 pneumoperitoneum(P>0.05). With time going on, ADM and CGRP level at T3 and T4 was remarkably lower than that at T1 (P<0.01);ADM and CGRP level at T5 would come to that at T1. Conclusions CO2 pneumoperitoneum during laparoscopic operation is the major factor causing decreased ADM and CGRP level;The decrease of ADM and CGRP level will influence the cardiac endocrine function,so CO2 pneumoperitoneum operation should be taken seri-ously when the patients have heart disease and cryptorrhea.

AIM To study the antiperoxide damage of injectio tanshin during dissolving thrombus to cure the acute myocardial infarction(AMI). METHODS AMI model formatted by electricity to irritate the left ventricular branch of coronary artery of open-chest rabbit,and observation aspects including electrocardiogram(ECG),cardiac output and contractility,serumal enzymatic activities and biochemistry and pathology of lipid peroxide extent of myocardial infarction zone. RESULTS By urinate kinase(UK) combining with injection tanshin,abnormal changes of ECG, descending of cardiac contractiles and cardiac output,going up of serum cardiac damage enzyme(CPK-MB and LDH),and the malondialdehyde(MDA)of serum and myocardial infarction zone could be remarkably reduced or avoided during UK dissolving the thrombus to cure AMI rabbits. CONCLUSION Injection tanshin can clearly reduce or avoid lipid peroxidation damage during UK dissolving the thrombus to cure the acute myocardial infarction.

Objective To screen and identify proteins that interact with receptor for advanced glycation end products (RAGE) with T7 select phage display system. Method GST-tagged fusion protein of RAGE was coated on glutathione suporflow resin and binding screens were performed with human lung T7 phage library. Results After 6 rounds of biopanning, 64 independent plaques were scraped and processed with EDTA. The inserted fragments in these plaques were amplified by PCR and the PCR products were purified by a gel recovery method. The sequences of the insertions were identified and analyzed with BLAST program in GeneBank. Twenty-five clones of encoding proteins were found. Conclusions T7 select phage display system is a convenient, rapid and effective method for screening binding proteins. This study provides an approach to explore complicated functions and novel mechanisms of RAGE-related signal transduction pathways.

Objective To compare the clinical effect of platelet rich plasma and sodium hyaluronate in the treatment of knee osteoarthritis .Methods According to the different treatment methods ,100 patients with knee osteo-arthritis were divided into the control group ( for treatment of platelet rich plasma ) and study group ( for treatment of sodium hyaluronate),each group 50 cases.The treatment effect would be compared between two groups ,and the patients were scored knee joint active function using Lysholms score standard ,using visual analogue score method on patients with knee joint pain .Results The excellent and good rate had no significant difference between the study group(98.0%) and control group(94.0%)(P>0.05);The knee joint activity of study group (76.81 ±9.24) was significantly better than that of the control group (60.46 ±7.41),the difference was statistically significant (t =-9.76,P<0.05);the knee joint pain degree of the study group (1.91 ±0.88) was lower than that of the control group (3.58 ±0.85),the difference was statistically significant (t=9.65,P<0.05).Conclusion Platelet rich plasma and the knee joint injection of sodium hyaluronate in the treatment of osteoarthritis were able to effectively alle -viate knee joint pain ,activity limitation and other clinical symptoms ,improve the curative rate ,but the treatment effect of sodium hyaluronate is more remarkable ,which is worthy of promotion .

NOK is a newly identified receptor protein-tyrosine kinase (PRTK) molecule that can promote tumorigenesis and metastasis. Previous data showed that NOK could activate the phosphatidylinositol 3'-kinase (PI3K) pathway in stable BaF3 cells. But how does NOK activate PI3K in cells remains unknown. It was showed that NOK physically interacted with the PI3K downstream effector Akt and enhanced its activation in human embryo kidney 293T (HEK293T) cells. Deletion mapping indicated that protein kinase B (Akt) was able to directly contact with the kinase domain of NOK. Inactivating the Akt kinase domain significantly reduced the intermolecular interaction between NOK and Akt, while constitutively active mutant of Akt apparently had a stronger interaction with NOK. NOK did not have an additive effect on insulin-mediated Akt activation. Overall, the results indicate that NOK might complex with Akt and directly activate PI3K/Akt signaling pathway.

Objective To assess the reliability of strength of shoulder rotator cuff using a hand-held dynamometer among subjects with spinal cord injury. Methods 20 spinal cord injury subjects without shoulder pain participated in this study. A hand-held dynamometer was used to assess the strength of shoulder rotator cuff. Two therapists (A/B) obtained the strength of shoulder rotator cuff. One of the therapists assessed again 5days later. The intra- and inter-reliability (ICC) of strength of shoulder rotator was calculated. Results The strength of rotator cuff was assessed by the hand-held dynamometer. The intra-reliability of external and internal muscle strength was 0.94 and 0.95, respectively. The inter-reliability was 0.91 and 0.93, respectively. Conclusion A hand-held dynamometer is a feasible and reliable tool to assess the strength of rotator cuff among subjects with spinal cord injury.

Objective:To provide a standard protocol for the rapid quantitative analysis of neutrophils in inflamed corneas with flow cytometry.Methods:The corneal epithelium layer of 15 C57BL/6 mice (6-8 weeks old) was mechanically scraped off using a golf-like knife to generate a 2 mm wound region.The mouse corneas with intact limbus were cut out at 18 hours after abrasion.After mechanical shredding, the single cell suspension was obtained by collagenase I and DNase digestion.Then, the number of neutrophils in the corneal cells was sorted under the FACSCanto flow cytometer using the gate technique.Another 6 mice were taken and randomized into wounded group and normal group according to a random number table method, with 3 mice in each group.Corneal cell staining was performed using fluorescent-conjugated anti-mouse CD45, Ly6G, and CD11b antibodies.The number of neutrophils in the corneas of the two groups were enumerated and compared.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO). The study protocol was approved by the Animal Ethics Committee of Medical College of Jinan University (No.JN-A-2002-01).Results:A standard procedure for detecting neutrophils in the cornea by flow cytometry was established.The ratio of CD45 + cells in the total corneal tissue cell population was (20.93±1.72)%.The Ly6G + and CD11b + double positive neutrophil population was sorted in the wounded corneal cell population.The ratios of Ly6G + and CD116 + cells in the CD45 + cells were (75.50±3.25)% and (93.40±4.53)%, respectively, and the ratio of the Ly6G + and CD11b + double positive neutrophils in the total number of CD45 + cells was (67.33±2.80)%.In addition, the number of neutrophils recruited to the cornea at 18 hours after corneal abrasion was (151.47±10.82)%, which was higher than (15.36±1.02)% in the normal cornea ( t=21.689, P<0.01). Conclusions:Flow cytometry can quickly and accurately quantitatively analyze the neutrophil population in the wounded cornea.It provides a rapid quantitative analysis method to further evaluate the changes of neutrophils in corneal inflammation caused by different reasons.

Objective To analyze the pathogen distribution and drug resistance of candida isolated from children with blood infections in our hospital,and to provide reference for clinical effective prevention and treatment.Methods The blood specimens of pediatric patients were collected between January 2009 and December 2015,and were cultured using BacT/ALERT 3D and BD9140 instruments.The candida were separated with Sobaurandps agar culture medium,and identified with chromogenic medium,API 20CAUX test strips or VITEK-2 compact YST card.The minimal inhibitory concentration of 5 drugs were determined by ATB FUNGUS 3 system.Results In 176 cases,92 strains (52.3％) were from neonatal ward,and 46 strains (26.1％) were from PICU.In newborn group,85 strains were isolated from premature,which contained the low and very low birth weight infants (37 strains),pneumonia(20 strains),neonatal respiratory distress syndrome(9 strains).In PICU,the strains were commonly isolated from children with severe infection.Among 176 strains of candida,71 strains (40.3％) were C.albicans,62 strains (35.2％) were C.parapsilosis,16 strains(9.1％) were C.glabrata,9 strains(5.1％) were C.tropicalis,and 18 strains(10.2％) belonged to other candida.Conclusion Candida blood infections can happen at all age of chlidren.The most common strains detected from blood were C.albicans,followed by C.parapsilosis.Most of these strains are susceptible to antifungal drugs,such as fluconazole,except C.glabrata.The sensitive rates to commonly used antifungal drug are more than 93％.The selection of antifungal drugs should be based on the species of strains.

Objective To study the clinical characteristic and antifungal drug sensitivity of neonatal Candida parapsilosis infections in blood and to provide reference for clinical prevention and therapy.Methods The drug resistance of Candida parapsilosis which were isolated from 8 neonatal blood specimen on the laboratory were analyzed and clinical characteristic of infections in neonates were investigated retrospectively.The blood cultured with BD9120.The fungi were isolated with Sabourand Dextrose Agar and CHROMagar colored medium and identified with API20C.The susceptibility test was then performed with FUNGUS3 micro dilution plate.Results Candida parapsilosis susceptibility results to antifungal drugs showed that 5-flucytosine≤ 4 μg/ml,amphotericin B ≤ 0.5 μg/ml,fluconazole ≤ 1 ～ 2.0 μg/ml,itraconazole ≤ 0.125 ～0.125 μg/ml,voriconazole ≤0.06 ～0.06 μg/ml.Seven cases were preterm infants(low birth weight infants or extremely low birth weight infants),one case was term infant after operation of congenital pyloric atresia.Before blood culture,all the 8 cases of Candida parapsilosis sepsis had received broad-spectrum antibiotics and intravenous nutrition.All the 8 cases had received peripherally inserted central catheter,and 3 cases had received mechanical ventilation.Four cases with fluconazole,3 cases with fluconazole and amphotericin B,1 case with fluconazole at the onset but changed VFend,all 8 cases were cured.Conclusion Candida parapsilosis has become the one of the main pathogens of neonatal infection fungal in blood of premature or low birth weight infants,which is sensitive to 5 kinds of antifungal drugs in vitro susceptibility test.Early detection and antifungal therapy can improve the prognosis.