Objective To evaluate the effect of collagen shield delivering fluconazole eye drops made by centrifugal method on treating fungal keratitis of rabbit eyes.Methods Using centrifugal method to make corneal collagen shield,after soaked with fluconazole eye drops,the collagen shields were used to treat fungal keratitis of rabbit eyes.Results The healing focus time of collagen shield delivering fluconazole eye drops group was average 11 days,the healing focus time of only delivering fluconazole eye drops group was average 18 days.The cure rate and effective rate of 2 groups were 78.94%,63.15% and 92.10%,78.94%,there were significance between 2 groups(P<0.05).Conclusion It was innovation for corneal collagen shield delivering fluconazole eye drops made by centrifugal method to cure fungal keratitis,which had broad development foreground.

Objective To evaluate the role of spinal sigma-1 receptors in the maintenance of bone cancer pain (BCP) in rats and the relationship with extracellular signal-regulated kinase (ERK).Methods Part Ⅰ Twenty-four female Sprague-Dawley rats,weighing 180-220 g,were randomized into 2 groups using a random number table:sham operation group (S group,n =4) and BCP group (n =20).BCP was induced by inoculating Walker 256 mammary gland carcinoma cells into the medullary cavity of the right tibia.Four rats were sacrificed on day 10 after inoculation in S group or on day 3,5,7,10 and 14 after inoculation in BCP group,and the L4-6 segments of the spinal cord were removed to measure the expression of sigma-1 receptors by Western blot.Part Ⅱ Forty female Sprague-Dawley rats,weighing 180-220 g,were randomized into 4 groups (n =10 each) using a random number table:sham operation group (S group),sigma-1 receptor inhibitor BD1047 group (BD group),BCP group,and BCP + BD1047 group (BCP + BD group).On day 10 to 14 after inoculation,normal saline 20 μl was injected intrathecally once a day in S and BCP groups,or BD1047 120 nmol/20μl was injected intrathecally once a day in BD and BCP + BD groups.Mechanical paw withdrawal threshold (MWT) to yon Frey filament stimulation was measured one day before inoculation,on day 3,5 and 7 after inoculation,and on day 10,12 and 14 after administration.After measurement of MWT on day 14 after inoculation,the rats were sacrificed and the L4-6 segments of the spinal cord were removed to determine the expression of phosphorylated ERK (p-ERK) by Western blot.Results Part Ⅰ Compared with group S,the expression of sigma-1 was significantly up-regulated and peaked on day 10 after operation in group BCP.Part Ⅱ Compared with S group,no significant changes were found in MWT and p-ERK expression at each time point in BD group,and MWT was decreased and p-ERK expression was up-regulated in BCP and BCP + BD groups.Compared with group BCP,after intrathecal injection of BD1047,MWT was significantly increased and the expression of p-ERK was down-regulated in BCP + BD group.Conclusion Spinal sigma-1 receptors are involved in the maintenance of BCP in rats possibly through promoting phosphorylation of ERK.

Objective To investigate the significance and effect of ultrasonic diagnosis on the autologous bone marrow mesenchymal stem cells (MSC) in angiogenesis. Methods Twenty-four New Zealand rabbits were divided into experiment group (12) and the control group (12). Then rabbit bone marrow MSCs from experiment group were isolated, caltured and marked with Brdu. After ischemic hind limb animal model on all rabbits was set up, autologous bone marrow MSCs were directly injected into the ischemic hind limb muscles in experiment group while same volume normal saline was used in the control group. Two weeks after the implantation of autologous bone marrow MSCs, 2D and color Doppler flow imaging (CDFI) detection were used in rabbit femoral artery of the two groups to observe the inner diameter of the blood vessel, the peak velocity and the acceleration time. The disposition of transplaned cells and the state of angiogenesis in ischemic muscles were assessed using immunofluorescence staining. Results The results of 2D and Doppler ultrasound detection showed the inner diameter of the blood vessel and the peak velocity of the blood current in experiment group obviously higher than that of the control group , and the acceleration time was obviously smaller than that of the control group P<0.01. The immunofluorescence staining showed there were transplanted cells existed in transplanted portion and state of angiogenesis was supurior obviously than that of the control. Conclusions Bone marrow MSCs had the effect to promote angiogenesis. Implantation of autologous bone marrow MSCs was a simple and efficient therapeutic method for the ischemia hind limb. Using high-frequency ultrasound to detect femoral artery may provide a practical and useful method to evaluate the effect on implanted autologous bone marrow mesenchymal stem cells.

By using enrichment media NPC and CVP, 792 strains of Pseudomonas and 515 strains of Erwinia were isolated from the rhizosphere of Digitaria adscendens (H. B. K.) Hem and Setaria vindis (L.) Beanv. Following which, experiments of antimetabolic test with E. coli, seed emergence controlling of S. viridis, herbicidal activity and security with green grass were carried out to select the desired bacteria. As the result, the selected strain, S7, could wholly control the seed emergence of S. viridis without any harm to the two tested green grass. And more, S7 promoted the seed emergence of Festuca arundinacea slightly. In spite of the comparatively low corrected mortality (56. 7%) of S7 after emergence of S. viridis, Selecting of microbial herbicides from weed DRB is thought to be more prospective.

Objective To explore the alteration and effect of autophagy in pancreas tissue of rat injected by exenatide.Methods Diabetes model rats were induced by two-month high-sugar and high-fat diet and streptozotocin injection (35 mg/kg) in normal rats.50 SD male rats were divided into four groups according to the principle of complete random design,namely normal control group (n=10),normal exenatide-injected group (n=10),diabetes-model control group (n=l5) and diabetes-model exenatide-injected group (n=15).Rats in exenatide-injected groups were subcutaneously injected with exenatide respectively in 5 μg/kg dose each time,twice a day,at 8 a.m.and 6 p.m.Animal weights were weighted weekly and the dose of exenatide was adjusted according to current weight.Rats in the two control groups were injected with the corresponding amount of saline.Mter 10 weeks of treatment,all rats were killed and pancreatic tissues were disposed.Immunohistochemistry was used to measure the expression of GLP-1R in pancreatic tissues.Western blot was used to test the expressions of LC3-Ⅰ,LC3-Ⅱ and p62 in pancreatic tissues,and LC3-Ⅱ/Ⅰ ratio and p62 were compared between any two groups.All specimens were stained with hematoxylin-eosin (HE).The data were expressed as means ± standard deviation and were analyzed by unpaired Student t test using SPSS 18.0 statistics software.P value ＜0.05 was considered to be statistically significant for all tests.Results The pancreatic tissues from 13 rats (6 from the normal exenatide-injected group and 7 from the diabetes-model exenatide-injected group) appeared pathological changes such as gland structure damage,pancreatic cells atrophy and cells compartment broadening.The expressions of GLP-1R,LC3-Ⅱ and p62,and LC3B-Ⅱ/Ⅰ ratio in the two exenatide-injected groups were higher than those in the respective control group,and the differences had statistical significance (P＜0.05).Conclusions Long-term subcutaneous injection of exenatide can upregulate the expression of GLP-1R in rat pancreatic acinar cells and may induce the impaiment of autophagy flux in rat pancreatic cell.

A 23-year-old man in remission from acute myeloblastic leukemia after allogeneic peripheral blood stem cell transplantation developed peripheral neuropathy presenting as sciatic and peroneal nerve deficits. Electrophysiological tests localized the lesions to the left sciatic and common peroneal nerve. Magnetic resonance imaging revealed nerve thickening and enhancement, while a positron emission tomography-computed tomography scan demonstrated increased fluorodeoxyglucose uptake tracking along the nerve, suggesting peripheral nerve infiltration. This report demonstrates an unusual presentation of acute leukemia relapse presenting as focal neuropathy

To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells. Methods:U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to de-tect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expres-sion. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regula-tion. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohisto-chemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections. Results: In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection. Conclusion:VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.

ObjectiveTo investigate the application of rapid immunohistochemical staining technique in intraoperative frozen section diagnosis of thyroid neoplasm.Methods MaxVision one-step rapid immunohistochemical staining technique was used to detect the expression of CK19,HBME-1,and Gal-3 in frozen section of papillary thyroid carcinoma(PTC)andthyroid benign lesions.MaxVision conventional immunohistochemistry of frozen remaining tissue was served as control.ResultsMaxVision one-step rapid immunohistochemical staining technique could be completed in 20 minutes.The positive localizations of three markers detected by rapid immunohistochemistry were similar to conventional immunohistochemistry, in general.The expression of CK19 was located in cytoplasm and cellular membrane.Gal-3 and HBME-1 were mainly detected in follicular luminal border and/or surface of papilla. The staining intensity in rapid immunohistochemistry was stronger than that in conventional immunohistochemistry. The positive rates of CK19,HBME-1,and Gal-3 by rapid immunohistochemistry in frozen sections were: 0 (0/28),10.7 ％ (3/28),0 (0/28),respectively,for benign lesions (nodular goiter,Hashimoto thyroiditis,thyroid adenoma); and 94.9 ％(37/39),92.3 ％ (36/39),92.3 ％ (36/39),respectively,for PTC.The expression of three markers between thyroid benign lesions and PTC had a significant difference (x2 =59.326,55.861,44.605,all P ＜ 0.001).In benign lesions,the rate of same case with two and more positive markers was 0,while in PTC it was 100 ％ and significantly different (x2 =67.000,P ＜ 0.05).ConclusionMaxVision one-step rapid immunohistochemical staining technique could be applied in intraoperative frozen section diagnosis.Detecting CK19,HBME-1,and Gal-3 expression in intraoperative frozen section has an auxiliary value for diagnosis of PTC.

Objective To study the correlation between time of fever onset in the course of patients'illness and etiologies of fever of unknown origin (FUO).Methods A total of 1 570 patients with FUO admitted from January 2013 to December 2014 were retrospectively analyzed, and clinical data ( sex, age, time of fever onset) of 348 patients meeting FUO diagnosis criteria with definite etiology diagnosis and time of fever onset were collected for multivariate logistic regression analysis after bias check.Results No statistically significant bias was found between 348 selected cases and 1 570 overall cases in gender (χ2 =0.029, P=0.903) and age (t=-1.040, P=0.299), and multivariate logistic regression analysis showed positive correlation between fever onset during 13: 00-18: 00 and infection (P=0.044, B=1.275), 18:00-24: 00 and connective tissue diseases ( P =0.029, B =0.838 ) , and showed negative correlation between age and miscellaneous (P =0.010, B =-0.042).Conclusions Characteristics of fever onset time may have significant value in preliminary diagnosis and guiding the correct direction of final definite diagnosis by means of targeted examinations or diagnostic treatments.It is worth to be further studied and discussed.

Objective To examine the expression and analyze the significance of autophagy-related gene microtubule-associated protein 1 light chain 3 (MAP1-LC3,LC3),p62 and lysosorne-associated membrane protein 2 (LAMP-2) in pancreatic tissues of mice with acute pancreatitis (AP).Methods Twenty mice were randomized into AP group and control group,and the number of mice was equal between two groups.AP group was intra-peritoneally injected by 20％ L-arginine solution (two injections of 4 g/kg body weight,every 1 h) in the dosage of 4 g/l kg twice every 1 hour to establish AP model,while control group was administered with equal volume of normal saline by intra-peritoneal injection.All the mice were euthanized at 24 hour after the last injection.Pancreatic histopathological changes were measured.In addition,the protein expressions of LC3,p62 and LAMP-2 were detected by Western blot.Results No obvious pathological changes were observed in control group.Pancreatic acinar edema,structure destruction,missing,the obvious widening of interlobular septum,small interlobular septum and acinar septum,and the necrosis of acinar cells at different degrees were observed in AP group.The pathological score for tissue edema,hemorrhage,necrosis and inflammation in AP group was 3.13 ± 0.50,2.83 ± 0.32,3.25 ± 0.46 and 3.16 ± 0.47,respectively,which was all 0 in control group.The differences were statistically significant between AP group and control group (P ＜ 0.01).In AP group,the ratio of LC3-Ⅱ/LC3-Ⅰ,p62 and LAMP-2 protein in pancreatic tissue were 1.16 ± 0.08,0.94 ± 0.04 and 0.35 ± 0.04,respectively,which were 0.24 ± 0.02,0.34 ± 0.03 and 0.95 ± 0.03 in control group.The ratio of LC3-Ⅱ/LC3-Ⅰ and p62 protein in pancreatic tissue in AP group were much higher than those in control group,while LAMP-2 in AP group was lower than that in control group,and there was statistically significant difference between two groups (all P ＜0.01).Conclusions Intraperitoneal injection of L-arginine could induce acute pancreatitis,and autophagy is impaired,which was associated with decreased LAMP-2 protein expression.