BACKGROUND:Previous experiments reported that basic fibroblast growth factor(bFGF) can change the shape of corneal endothelial cells,accelerate cell division and chemotaxis,and repair corneal endothelia cells injury in vivo and in vitro. OBJECTIVE:To investigate the effect of bFGF on cat corneal endothelial cells proliferation. DESIGN,TIME AND SETTING:The cell morphological observation was performed at the Central Laboratory of the Affiliated Hospital of Medical College of Qingdao University between January 2005 and December 2006. MATERIALS:Domestic cats were purchased from Experimental Animal Center of the Affiliated Hospital of Medical College of Qingdao University. METHODS:The corneal endothelial cells of cats were primarily cultured,and neurone specific enolase(NSE) immunocytochemistry staining was performed. After cell passage,bFGF(1,10,100 ?g/L) was added,continuously hatched for 1-5 days. MAIN OUTCOME MEASURES:MTT method was used to determine the proliferation of corneal endothelial cells;meantime,inverted phase contrast microscope and transmission electron microscope were used to observe the ultrastructure of cells. RESULTS:At days 1,3 and 5 after bFGF was added,the absorbance value in 490 nm was significantly higher than that of control group,especially in 10 ?g/L group. CONCLUSION:The bFGF can promote proliferation of cat corneal endothelial cells with effective concentration of 10 ?g/L .

BACKGROUND:Taurine is an important non-enzymatic system antioxidant in the lens.The mechanism of its anti-oxidative effect is mainly to protect lens from oxidative injury by anti-lipid peroxidation.OBJECTIVE:This study was to observe the effect of exogenous taurine on lens epithefial cell apoptosis-induced by H2O2 in vitro.DESIGN:A randomized controlled animal experiment.SETTING:Department of Ophthalmology,Affiliated Hospital of Qingdao University Medical College.MATERIALS:Seventy-five adult New Zealand standard tabbits,of either gender,weighing 1.5-2.5 kg,were provided by Qingdao Laboratory Animal Center.Reagent kit for in situ detecting cell apoptosis(Sigma Company,USA),taurine and H2O2(Shanghai Guangda Chemical Reagent Factory,China)were included in this study.METHODS:This study was performed in the Department of Ophthalmology and Central Laboratory.Affiliated Hospital of Qingdao University Medical College from Mav 2005 to June 2007.Rabbit lenses were harvested and randomly divided into 3 groups:control group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium which was renewed every 24 hours,H2O2 group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 with addition of 62 μL H202(30 g/L)every 6 hours,and H2O2+taurine group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 and 10 g/L taurine.which was renewed every 6 hours.The protocol was conducted in accordance with ethical guidelines for the use and care of animals.MAIN OUTCOME MEASURES:Observation of lens opacity 6.12,24.48 and 72 hours after culture;Lens epithelial cell apoptosis determined by DNA in situ end labeling and DNA fragment analysis.RESULTS:Lens opacity:The lens opacity in the H2O2 group was aggregrated gradually along with the time of oxidative injury.The lens opacity in the H2O2 group was severer than that in the H2O2+taurine group.Lens epithelial cell apoptosis:There were no apoptotic cells in the control group within 72 hours.The number of apoptotic cells in the H2O2 group was increased gradually with the prolonged time of oxidative injury.Till the 72nd hour,the cells were all tamed into apoptotic cells.A few aopototic celIs were found in the H2O2+taurine group since hour 24,and then were more and more,and the number of apoptotic cells accounted for about 30%at hour 72.The apoptotic rate in the H202+taurine group was significantly lower than that in the H2O group at each time point(q=8.6845,P＜0.01).There was no significant difference in apoptotic rate between the H202+taurine group and the control group (P＞0.05). Findings of DNA fragmentation assay:The DNA"ladder"was found in the H2O2 group at hours 24,36,48 and 72,while no DNA"ladder"but only normal electrophoresis straps were present in the other two groups 24 hours after culture.CONCLUSION:Taurine can inhibit the oxidative injury-induced apoptosis of rabbit lens epithelial cells,and alleviate lens opacity.

Objective:To observe influence of enhanced external counterpulsation (EECP) on plasma levels of lipo‐protein-associated phospholipase A2 (Lp‐PLA2) and hsCRP in patients with coronary heart disease (CHD) ,and preliminarily explore anti -inflammatory mechanism of EECP in prevention and treatment of CHD .Methods :A to‐tal of 85 CHD patients were selected from our hospital ,randomly divided into routine treatment group (n=42) and EECP group [n=43 ,received EECP based on routine treatment ,once/d ,60min each time ,continuous five weeks were regarded as one course (35 times)] .Plasma concentrations of Lp‐PLA2 and hsCRP were measured in all sub‐jects at the enrollment day and after five -week treatment .Results:Compared with before treatment ,after five-week treatment ,there were significant reductions in plasma levels of Lp‐PLA2 [(58.46 ± 40.04)μg/L vs .(33.94 ± 23.22)μg/L] and hsCRP [ (3.54 ± 2.22)μg/ml vs .(2.19 ± 1.16)μg/ml] in EECP group (P<0.01 both) ,and there were no significant difference in routine treatment group between before and after treatment .Compared with routine treatment group ,after five-week treatment ,there were significant reductions in plasma levels of Lp‐PLA2 [ (56.87 ± 33.69)μg/L vs .(33.94 ± 23.22)μg/L] and hsCRP [ (3.63 ± 1.60)μg/ml vs .(2.19 ± 1.16)μg/ml] in EECP group , P<0.01 both .Conclusion:1. EECP of one course can significantly reduce plasma concentrations of Lp‐PLA2 and hsCRP in CHD patients , indicating that anti - inflammatory may be one of its mechanism in preven‐tion and treatment of CHD .

Objective:To explore the relationship among plasma level of lipoprotein‐associated phospholipase A2 (Lp‐PLA2) ,ankle brachial index (ABI) ,brachial‐ankle pulse wave velocity (baPWV) and coronary heart disease (CHD) .Methods :According to results of coronary angiography ,a total of 120 patients with suspected or diagnosed CHD were divided into CHD group (n=90) and non‐CHD group (n=30) .CHD group was further divided into sin‐gle vessel coronary disease group (single vessel group ,n=30) ,double‐vessel coronary disease group (double‐vessel group ,n=30) and multi‐vessel coronary disease group (multi‐vessel group ,n= 30) .Plasma Lp‐PLA2 level ,ABI and baPWV were measured and compared among all groups .Results:Compared with non‐CHD group ,there were significant rise in plasma Lp‐PLA2 level [ (23.60 ± 13.33)μg/L vs .(36.65 ± 17.24)μg/L] and baPWV [ (1244.27 ± 127.85) cm/s vs .(1753.08 ± 284.32) cm/s] in CHD group ,P<0.01 both .In CHD group ,compared with single vessel group ,plasma Lp‐PLA2 level significantly rose [ (25.81 ± 8.97)μg/L vs .(35.03 ± 9.80)μg/L vs .(49.13 ± 21.22)μg/L] in double‐vessel group and multi‐vessel group ,and that of multi‐vessel group was significantly higher than that of double‐vessel group ( P<0.05 or <0.01 );baPWV significantly rose [ (1579.77 ± 178.05 ) cm/s vs . (1808.07 ± 272.11) cm/s ,(1871.40 ± 306.03) cm/s] in double‐vessel group and multi‐vessel group , P<0.01 both . ABI of single vessel group and double‐vessel group were significantly higher than that of multi‐vessle group [ (1.19 ± 0.08) ,(1.17 ± 0.07) vs .(1.11 ± 0.15)] ,P<0.01 or <0.05 .Conclusion:Plasma Lp‐PLA2 level ,ABI and baP‐WV are related to CHD and its lesion degree ,combined measurement of these three indexes can predict CHD and its severity more accurately ,then preventing and treating CHD should be more effective .

PurposeTo explore the value of transesophageal echocardiography (TEE) monitoring mini-incision transthoracic occlusion of ventricular septal defect (VSD) with asymmetric ventricular defects occlude in preoperative selection of patients, intraoperative guidance monitoring and postoperative evaluation.Materials and MethodsForty-five cases of VSD undertook mini-incision transthoracic occlusion with asymmetric ventricular defects occlude were enrolled, their clinical data was analyzed retrospectively, preoperative transthoracic echocardiography (TTE) was applied for choosing appropriate cases. During surgery, TEE was employed for evaluating the VSD and in which perimembranous VSD was found in 21 cases, intracristal VSD in 15 cases and subpulmonic VSD in 9 cases, suitable eccentric type occluders were chosen, guide occluder was placed, and the occlusion effect was evaluated right after operation.ResultsThe procedures were completed successfully in 42 cases, with a successful rate of 93.3%, of which 21 cases had perimembranous VSD, 15 cases had intracristal VSD, and 6 cases had subpulmonic VSD. The diameter of the VSD ranged from 3 to 7 mm, averaging (4.5±0.7) mm, the diameter of occluders ranged from 4 to 8 mm, averaging (5.7±1.2) mm, there was a good positive correlation between size of VSD and occlude (r=0.87,P<0.05). All patients received follow-ups from 3 months to 24 months after operation, all the occluders located normally, with no more than mildly residual shunt, valve regurgitation or severe arrhythmia discovered.ConclusionMini-incision transthoracic occlusion of ventricular septal defect (VSD) with asymmetric ventricular defects occlude has high success rates, minimal injury, and lower complication rate. TEE can play a vital role by improving the success rate and safety of surgery.

Inflammation plays an important role in occurrence and progression of atherosclerosis.Among numerous studies on inflammatory markers, the relationship between high sensitive C-reactive protein (hsCRP) and coronary heart disease (CHD) has been widely studied.The present article made a review about the relationship between hsCRP and CHD.

BACKGROUND: Oxidation injury can lead to the apoptosis of lens epithelial cells. Some researches have found that body supplement of vitamin C is beneficial to inhibit oxidation injury by increasing the level of vitamin C in aqueous humor.OBJECTIVE: To observe the inhibitive effect of vitamin C on H2O2 induced apoptosis of lens epithelial cell.DESIGN: Randomized controlled trial taking animal's lens as subject.SETTING: Department of Ophthalmology, Medical College of Qingdao University.MATERIALS: 120 adult New Zealand rabbits of both genders, weighing 3-5 kg, were selected. Vitamin C and 300 g/L H2O2 were purchased from Shanghai Guangda Chemical Reagent Factory.METHODS: The experiment was carried out in the Central Laboratory, the Affiliated Hospital of Medical College of Qingdao University from January 2002 to January 2004. ①Culture of lens: 240 lenses were isolat ed from 120 rabbits after being killed. Among them, 192 clear lenses were selected and divided randomly to three groups with 64 lenses in each group: control group, H2O2 group and H2O2+vitamin C group. Lenses in the control group were incubated in non-serum and non- phenolsulfonphthalein MEM medium, those in the H2O2 group incubated in non serum and non- phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 (62 μL of 30 g/L H2O2 was added into the medium every 6 hours to keep H2O2 level maintain 1 mmol/L in the medium), and those in the H2O2+vitamin C group incubated in non-serum and non- phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 and 1 mmol/L vitamin C. Under the same culture condition, the lenses in each group were detected at hours 6, 12, 18, 24, 36, 48, 72 and 108 after culture, respectively. ②Measurement of the opacity of lens: Under the white background, two black lines 0.5 mm in width, which were mutually vertical with an interval of 10 mm. The lenses were placed above the crossing to measure the opacity. ③Detecting the apoptosis of lens epithelial cells:The apoptosis of lens epithelial cells was examined using TUNEL method and DNA fragmentation assay under light microscope to calculate the apoptotic rate of epithelial cells.MAIN OUTCOME MEASURES: ①Opacity of the lenses in each group;②apoptotic rate of lens epithelial cells in each group.RESULTS: ①After 108 hours of culture, the opacity of the lenses in the H2O2+vitamin C group was obviously lower than that in the H2O2 group. ②The apoptotic rates in the control group were lower that those in the H2O2group at different time point and those in the H2O2+vitamin C groupat hours 24, 48 and 72 after culture(P＜0.05-0.01). The apoptotic rates in the H2O2+vitamin C group were significantly lower than those in the H2O2group at different time points after culture (P＜0.01). ③The findings of DNA fragment analysis showed that the DNA "ladder" was found in the H2O2 group during 24-72 hours, while no DNA "ladder" but only normal electrophoresis strap were present in the other two groups 24 hours after culture.CONCLUSION: Lens can take refuge from the oxidative injury of H2O2with the addition of vitamin C. As a result, the apoptosis rate of lens epithelial cells and further, cataract formation can be restrained.

Objective To investigate the mechanisms underlying intedeukin-22 (IL-22)-induced expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) in HaCaT cells.Methods Some HaCaT cells were divided into several inverention groups treated with IL-22 at concentrations of 12.5,25,50,100 μg/L,respectively and a control group treated with phosphate buffer saline (PBS).After 24-hour culture,total proteins were extracted from the HaCaT cells,and Western blot was performed to measure the expression of phosphorylated extracellular signalregulated kinase 1/2 (P-ERK1/2) in the mitogen-activated protein kinase (MAPK)-ERK1/2 pathway,as well as phosphorylated-JAK2 (P-JAK2) and phosphorylated-signal transducer and activator of transcription 3 (P-STAT3) in the JAK2/STAT3 pathway.In a blocking experiment,some HaCaT cells were divided into 4 groups to be treated with PBS,IL-22,PD98059 (an inhibitor of MAPK-ERK1/2) combined with IL-22 (PD98059 group),AG490 (an inhibitor of JAK2/STAT3) combined with IL-22 (AG490 group),respectively.After 24-hour treatment,total proteins and mRNAs were extracted from the HaCaT cells followed by Western blot and real-time quantitative reverse transcription-PCR for the measurement of protein and mRNA expressions of HB-EGF respectively.Statistical analysis was carried out with the software SPSS 16.0 by one-way analysis of variance (ANOVA) for intergroup comparisons and by Bonferroni's test for multiple comparisons.Results After treatment with IL-22 at the above 4 concentrations,the expressions of P-ERK1/2,P-JAK2 and P-STAT3 in HaCaT cells were all increased compared with the control group (all P ＜ 0.05).The protein and mRNA expression levels (expressed as the HB-EGF/β-actin ratio and 2-△△Cr respectively) of HB-EGF were both significantly decreased in the PD98059 group and AG490 group than in the IL-22 group (protein:0.183 ± 0.020 and 0.199 ± 0.011 vs.0.924 ± 0.032,F =37.700,36.400,respectively,both P ＜ 0.05; mRNA:1.034 ± 0.072 and 0.989 ± 0.038 vs.1.844 ± 0.135,F =11.271,13.429,respectively,both P ＜ 0.05).Conclusions IL-22 can activate the MAPK-ERK1/2 and JAK2/STAT3 signaling pathways in HaCaT cells,which may contribute to IL-22-induced expression of HB-EGF in HaCaT cells.

Objective To investigate the role of individual dental stent on taste protection in the patients with primary nasopharyngeal carcinoma.Methods Firstly,dosimetric evaluation of the tongue under the circumstance of 10 pairs of patient wearing dental stent or not was done.Secondly,a total of 61 patients were randomized into two arms:29 were in the trial arm and 32 in the control arm.The patients wore dental stent during radiotherapy in trial group,but not in control group.The endpoint was taste dysfunction before radiotherapy,every week during radiotherapy till the completion of radiotherapy,and followed up six months after radiotherapy.Results The mean dose of tip of tongue,body of tongue,base of tongue for patient with dental stent or not were (394.43±24.01) cGy,(986.35±77.15) cGy,(4 401.16±179.01) cGy and (677.03± 110.05) cGy,(2 418.19±414.18) cGy,(4 878.67±387.45) cGy (P value were 0.031,0.007 and 0.284,respectively) separately.By the completion of the radiotherapy,there were 9 (31.03 ％) patients suffering from a taste dysfunction in the trial group and 25 (78.13 ％) in the control group (P ＜ 0.001).Conclusions Individual dental stent has a potential tendency to relieve the taste impairment by reducing the irradiation dose of tongue.

Objective To investigate the specific expression of endoplasmic reticulum stress-related GRP78 and CHOP in podocyte of MKR mice with diabetic nephropathy (DN);To discuss the intervention effect of Zuogui Jiangtang Yishen Decoction (ZGJTYS). Methods 8-week old MKR mice were randomly divided into five groups:blank group, model group, ZGJTYS group, 4-Phenylbutyric acid group, gliquidone+benazepril group, and normal control group consisting of wild type C57BC/6 mice, 10 mice per group. All mice from model group and each treatment group received high-fat diet feed and unilateral nephrectomy to make the DN model. Mice from treatment groups received medicine intervention for four weeks. The levels of FBG were detected by electrochemical detection method, and the UmAlb was detected by ELISA. The expressions of GRP78, CHOP mRNA, and protein were detected by RT-PCR and Western blot. The morphological structure changes of the podocytes were observed by transmission electron microscopes. Results Compared with blank group, FBG and UmAlb in the model group increased significantly (P<0.01);the expressions of GRP78 mRNA and protein markedly decreased (P<0.01);the expressions of CHOP mRNA and protein increased (P<0.01). Compared with the model group, FBG and UmAlb in each treatment group significantly decreased (P<0.01);the expressions of GRP78 mRNA and protein in the ZGJTYS group and 4-Phenylbutyric acid group increased (P<0.01), the expressions of CHOP mRNA and protein decreased (P<0.01). The renal pathological lesions of each treatment group were significantly improved compared with model group. Conclusion ZGJTYS Decoction can effectively improve the damaged podocyte induced by endoplasmic reticulum stress, delay DN progress.