AIM: To study the infiltration of polymorphonuclear neutrophils ( PMNs ) after conjunctival flap covering in alkali-burned cornea.
●METHODS: Rabbit cornea alkali-burned model was made, then 50 rabbits were randomly divided into the experimental group ( n=25 ) and the control group ( n=25 ) . At the same time the surgery of conjunctival flap covering was given to rabbits of the experimental group. The condition developing of alkali-burned cornea was observed by slit lamp biomicroscopy, and took photos in two groups. The infiltration of PMNs was identified by hematoxylin eosin ( HE) staining in different periods.
●RESULTS:The quantity of PMNs increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. PMNs level of the experimental group was significantly lower than that in the control group, and the difference of 3, 14 and 21d was significant (P<0. 05).
●CONCLUSION: During the wound healing process, alkali - burned cornea has close relation with the infiltration of PMNs. The treatment of conjunctival flap covering for the severe alkali-burned cornea was found to have good effect.

Bacteremia ; diagnosis ; epidemiology ; microbiology ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Pseudomonas Infections ; diagnosis ; epidemiology ; Pseudomonas aeruginosa ; Retrospective Studies ; Rural Population ; Urban Population

Objective To investigate the action mechanism of glutamate-mediated signaling pathway in malignant melanoma. Methods WM451LU melanoma cells in log phase were classified into 6 groups, negative control group treated with PBS (100 μl), MK801 group treated with the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (100 μmol/L), CPCCOEt group treated with non-competitive metabotropic glutamate receptor 1 (mGluR1) antagonist CPCCOEt, MAP2 group transfected with adenovirus vector containing microtubule associated protein 2a (Ad-MAP2a), MK801 + MAP2 group treated with MK801 of 100 μmol/L and transfected with Ad-MAP2a, CPCCOEt + MAP2 group treated with CPCCOEt of 10 μmol/L and transfected with Ad-MAP2a. Western blot was performed to detect the expression of an ionotropic glutamate receptor, i.e., N-methyl-D-aspartate receptor type 2A (NMDAR2A) in WM451LU cells transfected with Ad-MAP2a. Scratch motility assay and cell invasion assay were conducted in vitro to detect the changes in migration and invasion ability of WM451LU cells after treated with Ad-MAP2a, MK-801, CPCCOEt alone or in combination. In vivo study was carried out to compare the inhibitory effect of the above treatments on melanoma. Results Western blot revealed a decrease in the expression of NMDAR2A in WM451LU cells after transfected with Ad-MAP2a. The scratch motility assay showed that the number of migrating cells per high power field was 117.04 ± 2.76 in MAP2 group,107.64 ± 6.50 in MK801 group,97.36 ± 4.79 in CPCCOEt group, 43.28 ± 3.02 in MK801 + MAP2 group,30.76 ± 3.97 in CPCCOEt + MAP2 group,significantly different from that in the negative control group (152.3 ± 5.75,all P ＜ 0.01 ). Cell invasion assay demonstrated that the average number of invading cells per high power field in the negative control was significantly higher than that in MAP2 group, MK801 group, CPCCOEt group, MK801+MAP2 group and CPCCOEt + MAP2 group (170.43 ±8.72 vs. 98.26 ± 3.84, 97.22 ± 5.54, 112.23 ± 7.21, 42.89 ± 5.06, 58.25 ± 6.68, P ＜ 0.05, 0.05, 0.05, 0.01and 0.01, respectively).A significant decrease was observed in the average volume of experimental melanoma in mice of MAP2 group, MK801 group, MK801 + MAP2 group, CPCCOEt group and CPCCOEt + MAP2 group compared with the negative control group (224.02 ± 46.19 mm3, 160.33 ± 33.91 mm3, 91.49 ± 21.48 mm3,202.30 ± 52.37 mm3, 111.13 ± 69.81 mm3 vs. 342.70 ± 60.92 mm3, all P ＜ 0.01 ). Conclusions To block the glutamate signaling pathway in vitro can inhibit the invasion and migration of melanoma cells, and to block the pathway in vivo can inhibit the growth of malignant melanoma and alter the morphology of melanoma cells.

Objective:To evaluate the application method and effect of standardized scenario simulation teaching based on Kirkpatrick model in vocational protection education for nursing students.Methods:A historical controlled trial study was designed. Practical nursing students enrolled in 2018-2019 and 2017-2018 were selected into the experimental group ( n=203) and control group ( n=196), respectively. The experimental group adopted standardized scenario simulation teaching in the prevention and control education of needlestick injuries, and the control group adopted traditional classroom lecture. Using the Kirkpatrick model, the teaching effect of needlestick injuries protection for nursing students were compared between the 2 groups from such 4 levels as in reaction level, learning level, behavior level and results level. Results:There was no significant difference in the baseline data between the two groups in terms of age, gender, educational background and test scores of nursing professional knowledge as compared to that before practice. In reaction level: the nursing students' satisfaction of experimental group in teaching methods ( t=25.149, P<0.001) and teaching environment ( t=12.827, P<0.001) are higher than that of the control group, with statistical significance. In learning level: the test scores of needlestick injury knowledge in experimental group were significantly higher than those in control group ( t=8.221, P<0.001). In behavior level: the level of needlestick injury protection behavior in the experimental group was significantly higher than that in the control group ( t=9.250, P<0.001), and the knowledge conversion rate in the experimental group was higher than that in the control group ( t=6.054, P<0.001). In results level: the needlestick injuries incidence of experimental group was significantly lower than that of the control group ( χ2=15.815, P<0.001), the reported rate of needlestick injuries of experimental group was significantly lower than that of the control group ( χ2=14.185, P<0.001). Conclusion:The implementation of standardized scenario simulation teaching can effectively improve the effectiveness of vocational protection learning and reduce the incidence of needlestick injuries.

Objective To establish a zebrafish IGFBP-2 gene knock-down model by morphilino modified antisense oligonucleotide injection, so as to investigate the abnormal phenotypes of heart and vessels in early stage of zebrafish development and the expression of zebrafish cardiogenesis related genes. Methods The spatiotemporal expression of IGFBP-2 gene in early stage of zebrafish development was testified by whole mount in situ hybridization with antisense RNA IGFBP-2 probe. The IGFBP-2 morpholino (IGFBP-2 MO) that especially inhibited the gene promoter and standard control morpholino (Con-MO) were designed and synthesized by Gene-tools Corporation. Four different concentration gradients (0.05, 0.10, 0.25 and 1.0 mmol/L) were set as IGFBP-MO injection groups with 0.25 mmol/L Con-MO injection group and wild type group as controls. Contribution to the incidence of heart abnormal phenotypes and mortality rate induced by 4 different IGFBP-2 concentrations injection group was recorded and compared with 2 control groups. Heart abnormal phenotypes at different developmental stages in 0.25 mmol/L IGFBP-2 injection group were observed in detail. To validate the effectiveness of IGFBP-2 MO, the expression of enhanced green fluorescence presented by wild type zebrafish embryos at 12hpf which received single injection of IGFBP-2 EGFP recombinant plasmid and those co-injected with Con-MO or IGFBP-2 MO were detected. To investigate the regulation relationship between IGFBP-2 gene and other cardiogenesis related genes, expression of atrium specific marker gene Amhc was detected in IGFBP-2 MO and wild type group by in situ hybridization. Ventricle specific green fluorescence of Vmhc-EGFP transgenic zebrafish embryos whose IGFBP-2 gene was knocked-down were compared with those untreated. Zebrafish peripheral vascular development in the IGFBP-2 MO group was also checked out by micro-angiography. Results Whole mount in situ hybridization revealed that IGFBP-2 gene expressed in turn at eyes, midbrain and then focused on liver in early stage of zebrafish development. The micro-injection of 0.25 mmol/L IGFBP-2 MO resulted in heart malformation in nearly 60% of all injected zebrafish embryos. Heart malformation phenotypes included slow heart beat, pericardial edema, weak ventricle systole contraction and heart tube looping disorder. Some of them represented atria dilation, blood regurgitation and ciculation obstruction. Wild type zebrafish embryos that received single injection of IGFBP-2 EGFP plasmid DNA or co-injected with Con-MO presented strong enhanced green fluorescence at 12hpf, meanwhile, the fluorescence was barely seen in the embryos co-injected with IGFBP-2 MO. This strongly validated the gene specific knock-down effect of IGFBP-2 MO. Amhc was down-regulated at 48hpf in IGFBP-2 MO group. Vmhc-EGFP transgenic zebrafish down-regulated by IGFBP-2 gene also resulted in attenuated expression of ventriclar-specific green fluorescence protein at 48hpf. Intersegmental blood vessels of IGFBP-2 MO group by micro-angiography at 60hpf demonstrated an sparsate and chaos image, which suggested that IGFBP-2 gene expression was involved in the regulation of normal vascular development. Conclusions Micro-injection of IGFBP-2 MO is an efficient way to knock-down IGFBP-2 gene in zebrafish embryos. IGFBP-2 gene expression down-regulation leads to heart and vessels maldevelopment and have an impact on the expression of cardiogenesis related genes of zebrafish embryos as well. In short, IGFBP-2 plays a critical role in the normal cardiovascular development of zebrafish embryos.

The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.
China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Mass Spectrometry ; Quality Control ; Rhizome ; chemistry

Objective To investigate the roles of glutamate signaling pathway in the activation of human peripheral blood lymphocytes(PBLs) and pathogenesis of vitiligo.Methods Peripheral blood lymphocytes (PBLs) isolated from 5 patients with generalized vitiligo and 5 healthy controls were cultured in vitro.Flow cytometry was performed to quantify the expression of CD25 and interferon-γ on PBLs derived from healthy controls and treated with MK801 (a non-competitive antagonist of N-methyl-D-aspartic acid receptor,NMDAR) at 100 μmol/L or phosphate buffered saline (PBS) for 48 hours,as well as the level of reactive oxygen species (ROS) in the controlderived PBLs treated with MK801 at 100 μmol/L,NMDA (an agonist of N-methyl-D-aspartic acid receptor) at 0.5 mmol/L or PBS for 48 hours.The protein and mRNA expressions of NMDAR1 and NMDAR2A were measured by flow cytometry and real-time PCR,respectively,in PBLs from the healthy controls and vitiligo patients.Immunohistochemistry was used to observe the expressions of NMDAR1 and NMDAR2A in tissue specimens from depigmented and postinflammatory hyperpigmentation lesions of the patients with vitiligo and from normal skin of the healthy controls.Results Compared with the PBS-treated PBLs from the healthy controls,the MK801-treated PBLs showed a downregulated expression of CD25 (7.28％ ± 0.18％ vs.16.02％ ± 0.42％,P ＜ 0.01),but an upregulated proportion of CD25+IFN-γ+ lymphocytes (1.79％ ± 0.09％ vs.0.78％ ± 0.06％,P ＜ 0.01),and the NMDA-treated PBLs displayed a higher ROS level (101.1 ± 3.50 vs.69.80 ± 2.08,P＜ 0.01 ).The protein expression of NMDAR1 in PBLs was significantly higher in vitiligo patients than in the healthy controls (3.85 ± 2.17 vs.0.97 ±0.55,P ＜ 0.05).Conclusion Glutamate signaling pathway may be involved in the immunopathogenesis of vitiligo via affecting the secretion of interferon-γ by,and ROS level in,activated lymphocytes.

Bacterial N-Acyl-L-homoserine lactones (AHLs)-mediated quorum sensing is involved in the regulation of diverse biological functions. As the bacterial pathogen population density increases, AHLs concentration secreted by pathogen reaches a threshold and then interacts with their intercellular receptor and triggers expression of virulence genes. It is a promising approach to biologically control bacterial disease in plants and animals by manipulating bacterial AHL-quorum sensing with AHLs-degrading enzyme and AHL analogue.

Ralstonia metallidurans CH34 was isolated from the deposit of a znic factory .The degradation of phenol by R .metallidurans CH34 was investigated. The results showed that R . metallidurans CH34 possesses high ability to degrade phenol with the biodegradation rate constant of 0.33 . The optimal pH , temperature and volume of medium for phenol degradation are pH 7.0 , 30℃ , and 20%(v/v), respectively . In addition , this strain retains its ability to degrade phenol in the presence of high concentration of heavy metal ion .The sodium citrate , sodium succinate can enhance the degradation of phenol.

Objective The aim of this study was to introduce the multi-slate Markov model for the prediction of mild cognitive impairment (MCI) to Alzheimer' s disease (AD) and to find out the related factors for AD prevention and early intervention among the elderly.Methods MCI,moderate to severe cognitive impairment,and AD were defined as state 1,2 and 3,respectively.A three-state homogeneous model with discrete states and discrete times from data on six follow-up visits was constructed to explore factors for various progressive stages from MCI to AD.Transition probability and survival curve were made after the model fit assessment.Results At the level of 0.05,data from the multivariate analysis showed that gender (HR=I.23,95％CI:1.12-1.38),age (HR=I.37,95％ CI:1.07-1.72),hypertension (HR=l.54,95％ CI:1.31-2.19) were statistically significant for the transition from state 1 to state 2,while age (HR=0.78,95％ CI:0.69-0.98),education level (HR=1.35,95％ CI:1.09-1.86) and reading (HR=1.20,95％ CI:1.01-1.41 ) were statistically significant for transition from state 2 to state 1,and gender (HR=1.59,95％ CI:1.33-1.89),age (HR=1.33,95％ CI:1.02-1.64),hypertension (HR=l.22,95％ CI:1.11-1.43),diabetes (HR=1.52,95％CI:1.12-2.00),ApoEε4 (HR=1.44,95％CI:1.09-1.68) were statistically significant for transition from state 2 to state 3.Based on the fired model,the three-year transition probabilities during each state at average covariate level were estimated.Conclusion To delay the disease progression of MCI,phase by phase prevention measures could be adopted based on the main factors of each stage.Multi-state Markov model could imitate the natural history of disease and showed great advantage in dynamically evaluating the development of chronic diseases with multi-states and multi-faetors.