Background Animal model of fungal keratitis is an available tool to the experimental study of the pathogenesis mechanism of fungal keratitis. Current modeling methods of fungal keratitis include corneal scratching, corneal stroma injection and corneal surface lens methods. But these methods still have their own shortages. Objective This experiment was to create a fungal keratitis animal model by modifying corneal surface lens method. Methods Modified animal models of fungal keratitis were created by modified corneal surface lens method in 12 general adult New Zealand white rabbits. The filter papers soaked 108 spores / ml or A106spores / ml of spergillus fumigatus suspension were attached on the de-epithelial cornea surface and fixed with contact lens and tarsorrhaphy for 2 days, and the filter paper with physiological saline was used as control group. The symptoms of anterior segment were examined under the slit lamp in 3 ,7 and 14 days after surgery and scored based on the criteria of Dong. Corneal scraping was stained with 10% potassium hydroxide and calcofluor white stain to observed mycelium under the fluorescence microscope. Corneal tissue sections were examined by hematoxylin-eosin staining and periodic acid Schiff staining under the light microscope. The use of animal followed the Standard of Association for Research in Vision and Ophthalmology. Results Fungal keratitis models were successfully established in 6 eyes and 4 eyes in 108 spores/ml group (6/6) and 106 spores/ml group respectively. The symptom was more severer and score was higher in the eyes of 108 spores/ml group than that in 106 spores/ml group. At 3 and 7 days after surgery,the symptom scores of fungal keratitis models were higher than those of control group from 3 through 7 days with the statistically significant difference (P<0. 01) and the symptom scores of 108 spores/ml group were significantly higher than those of 106 spores/ml group (P<0. 01). At 14 days after surgery, the symptom scores of 108 spores/ml group were still higher than those of control group (P<0. 05). Fungal hyphae was seen in the corneal scrapes in 108 spores/ml group and 106 spores/ml group respectively from 3 through 7 days after surgery. Inflammatory cell infiltration, stroma cells necrosis and fungal hyphae were presented in 108 spores/ml group, and the corneal neovascularization could be observed in 108 spores / ml group 14 days later. Fungal culture revealed the positive outcome in both 3 and 7 days after surgery in 108 spores/ml group,but in 106 spores/ml group,the positive result was only in the 3rd day. Conclusion Modified corneal surface lens method is more feasible and sample in the model of Aspergillus keratitis. This animal model of Aspergillus keratitis is practical for the further study of fungal keratitis.

To investigate the invasive ability of the residual tumor cells after immunotherapy and explore the feasible approach suppressing the invasion, mice were inoculated with B16 cells, and then treated by gene therapy with p4-1BBL/psPD-1 or IFN-gamma. The production and activities of MMP-9 and MMP-2 in residual tumor tissues were analyzed with gelatin zymography 1 day and 7 days after the termination of the immunotherapy. The production of MMP-9 and MMP-2 by B16 cells treated with IFN-gamma was also analyzed. IFN-gamma-treated B16 cells were inoculated to mice via subcutaneous injection. The invasion of tumor to muscular tissue was analyzed. Gene therapy with CH50 was used to suppress the invasive growth of tumor. The results showed that the expression and the activities of MMP-9 and MMP-2 were significantly increased 7 days after the end of immunotherapy. The response of tumor cells to ECM molecules was intensified after the removal of IFN-gamma, resulting in significant increase of both the production and activities of MMP-9 and MMP-2, and the increased invasion of tumor. Gene therapy with CH50 effectively suppressed the invasive growth of tumor. It is concluded that the termination of immunotherapy may result in a higher metastatic potential of residual tumor cells. Suppressing tumor invasion by suitable treatment will improve the efficacy of immunotherapy.

Objective To investigate network user's attention rate of the keyword“cardiovascular and cerebrovascular diseases”(CCD). Methods The keyword was analyzed by using Baidu index. Results The overall trend of keyword search rose slightly. The mobile search grew significantly. During the Spring Festival overall search decreased significantly. According to clustering analysis the word“symptom”was most often used with“CCD”;30 to 39 years old age group accounted for about 50%of total number of online search. Men accounted for 70% of all keyword search, women accounted for 30%. The rankings of CCD search number were more on top in the economically developed area. Conclusion It is necessary to strengthen publicity and education to prevent the happening of CCD during Spring Festival. Men need to raise awareness about CCD. The construction of rural informatization should be speeded up. Network education could be used as a part of method for the primary and secondary prevention of CCD. Keyword analysis method could effectively analyze the network user search behavior and find information they need.

Objective To develop a BP26 recombinant BCG (rBCG-BP26) vaccine,and to observe the effects of rBCG-BP26 on CD4+,CD8+ T cells in immunized mice.Methods The recombinant shuttle vector pMV261-Ag85B-BP26 was constructed by using traditional molecular biological technology.The recombinant strains were obtained by kanamycin resistance screening and PCR identification after electroporation.Western blotting was used to detect the expression of recombinant BP26 vaccine in immunized mice.Safety experiment was carried out in three different groups:the target experiment(rBCG-BP26) group,the positive control(BCG) group and the negative control(PBS) group,15 BALB/c mice in each group.Intradermal inoculations of 100 μl rBCG-BP26 [containing 106 colony forming units(CFU)],BCG,and PBS were carried out,respectively.Signs of mice in each group were observed.After immunization for 10,20,30,and 40 days,body weight was weighed,and tail blood was collected to observe the change of peripheral blood CD4+ and CD8+ T cells by flow cytometry.Results The rBCG-BP26 was successfully constructed.The expression of BP26 protein was detected in the liquid medium and the bacteria cells.The results of safety test analysis showed that there were no significant differences in signs and body weights(F=2.468,0.331,1.520,0.739,all P＞ 0.05),between PBS group[ (19.24 ± 0.54),(21.37 ± 0.66),(22.83 ± 0.62),(25.06 ± 0.37)g],BCG group[ (19.90 ± 0.02),(21.53 ± 1.57),(21.95 ± 0.55),(24.70 ± 0.39)g]and rBCG-BP26 group[ (19.16 ± 0.55 ),(20.89 ± 0.20),(22.15 ± 0.76),(24.60 ± 0.64)g].The results of flow cytometry showed that the percentages of CD4+ T cell level were lower in BCG group(26.70％,33.07％) and rBCG-BP26 group( 13.40％,26.70％) than that of the PBS group(33.85％,29.33％) and the values of CD4+/CD8+ T cells increased in rBCG-BP26 group (0.69％,1.27％,1.57％,1.70％ ) 10,20 and 30 days after immunization.Conclusions Recombinant BCG-BP26 vaccine strain can express brucella BP26 protein efficiently.Furthermore,its virulence is mild,and it can activate CD4+,CD8+ T cells in the body.It can be used as one of candidate vaccine strain against brucellosis.

BACKGROUND:This systematic review aims to investigate the prediction value of diffusion tensor imaging for motor function recovery of ischemic stroke patients. METHODS:Cochrane Central Register of Controlled Trials (CENTRAL) (the Cochrane Library 2016, Issue 9), PubMed, Embase, Clarivate Analytics, Scopus, CINAHL, Chinese Biomedical Literature Database, China National Knowledge Infrastructure and Google Scholar were searched for either motor recovery or corticospinal tract integrity by diffusion tensor imaging in different stroke phase from January 1, 1970, to October 31, 2016. The study design and participants were subjected to metrological analysis. Correlation coefficient (r) was used for evaluating the relationship between fractional anisotropy (FA) and motor function outcome. Correlation coefficient values were extracted from each study, and 95％ confidence intervals (CIs) were calculated by Fisher's z transformation. Meta-analysis was conducted by STATA software. RESULTS:Fifteen studies with a total of 414 patients were included. Meta-analysis showed that FA in the subacute phase had the significant correlation with motor function outcome (ES=0.75, 95％CI 0.62-0.87), which showed moderate quality based on GRADE system. The weight correlation coefficient revealed that an effect size (ES) of FA in acute phase and chronic phase was 0.51 (95％CI 0.33-0.68) and 0.62 (95％CI 0.47-0.77) respectively. CONCLUSION:This meta-analysis reveals that FA in the subacute phase after ischemic stroke is a good predictor for functional motor recovery, which shows moderate quality based on the GRADE system.

AIM:To study whether inhibition of forkhead box protein M1(FoxM1) sensitizes leukemia K562 cells to homoharringtonine ( HHT ) .METHODS: K562 cells were incubated with HHT at different concentrations ( 0μmol/L, 0.015 μmol/L, 0.030μmol/L and 0.045μmol/L) for different time (0 h, 24 h, 48 h and 72 h).The mRNA and protein levels of FoxM1 were detected by real-time PCR and Western blot.FoxM1 siRNA was transfected into K562 cells with 0.015μmol/L HHT after 6 h.After 72 h incubation, the cell proliferation was detected by cell counting and soft agar assay, and the proportion of apoptotic K562 cells was determined by flow cytometry.The expression of c-Myc and Sp1 were detected by real-time PCR and Western blot.RESULTS:FoxM1 expression was reduced time-dependently and dose-dependently, suggesting that HHT mediated the downregulation of FoxM1 in K562 cells.In K562 cells, treatment with FoxM1 siRNA and HHT inhibited the cell proliferation and promoted the apoptosis significantly.Therefore, inhibition of FoxM1 sensitized leukemia K562 cells to HHT.The expression of c-Myc and Sp1 was positively regulated by FoxM1. CONCLUSION:HHT inhibits Forkhead box protein M1 expression in K562 cells.Inhibition of FoxM1 sensitizes K562 cells to HHT.

Objective To explore the clinical significance of sCD14-ST in diagnosing children sepsis and monitoring the treatment effects. Methods Case-control study. Patients were recruited by Longyan First Hospital from August 2013 to March 2015.The sCD14-ST, WBC, CRP, PCT and APACHE-Ⅱlevels were measured in 237 septic children ( infectious SIRS, including 120 bacterial and 117 nonbacterial SIRS) , 89 non-infectious SIRS and 156 healthy children.The changes and the correlation of the five indicators in septic children before and after treatment was compared.The diagnosis value of sCD14-ST in sepsis by the receiver operationg characteristic curves ( ROC) was analyzed.Results The sCD14-ST level in septic children [643.47(596.47 -690.46) ng/L] was higher than that in non-infectious SIRS [246.94(234.85-259.03)ng/L]and healthy control [151.00(142.79-159.22)ng/L](χ2 =121.850, 325.663, P<0.01).The whole blood level of sCD14-ST in bacterial sepsis [606.17(542.71-669.63) ng/L]and non-bacterial sepsis [679.83(610.37-747.29)ng/L]were no significance (χ2 =0.854, P=0.335), while compared with the control group, they had significant differences (χ2 =326.228, P<0.01) .Totally 80 blood culture samples were positive in 117 bacterial septic children, and the sCD14-ST level was not significant between Gram positive bacteria infection and [641.07 (553.82 -728.31) ng/L] Gram negative bacteria infection[750.00(597.10 -902.89) ng/L] (χ2 =1.12,P=0.29), but the sCD14-ST level in blood culture positive children was significantly higher than healthy controls ( chi-square =117. 46, 155.846, P<0.01).sCD14-ST, WBC, CRP, PCT, APACHE-Ⅱscore before and after treatment were all significantly decreased ( χ2 =44.569, 113.337, 63.986, 100.055, 51.015, P<0.01).The sCD14-ST was significantly positively related withCRP and APACHE-Ⅱ score before treatment, but there were no correlation with WBC and PCT.The ROC-AUC of sCD14-ST in septic children was 0.901.Compared with the ROC-AUCs of WBC ( 0.875 ) and CRP ( 0.836 ) , there were statistically significant different ( P<0.01).The sensitivity and specificity of sCD14-ST were 85%, 90%, respectively.Conclusions The study suggested that sCD14-ST superior value to the diagnosis of sepsis in children than other parameters.It should be applied as a valuable biomarker for early diagnosis and the evaluation of severity of sepsis.

OBJECTIVETo observe the dose-effect relationship of electroacupuncture of different current intensities combined with Morphine of different dosage on alleviating the rats' tibial cancer pain, and explore the possible mechanism, which could provide the experiment basis for alleviating the tibial cancer pain by electroacupuncture combined with Morphine.

METHODSOne hundred female Wistar rats were randomly divided into a normal group, a model group and eight treatment groups, 10 cases in each group. The rats in the treatment groups were treated by combined therapies of electroacupuncture of different intensities with 2 Hz /100 Hz dense-disperse wave on "Jiaji"(EX-B 2)and different dosage Morphine in 2 factor 3 level conditions, once a day for 6 days. The pain thresholds were observed before the treatment and 0 min, 1 h, 2 h and 5 h after the first treatment as well as after 3 and 6 times of treatments. The glial fibrillary acidic protein (GFAP) expression was determined by immunohistochemical method.

RESULTSThe rats' pain thresholds were significantly increased with electroacupuncture of 2 mA and 1 mA (all P < 0.01) on the 0 min, 1 h and 2 h of the first treatment, between which there were no significant differences (all P > 0.05). The pain threshold was still increased by electroacupuncture of 2 mA on the 5 h of the treatment (P < 0.01), while that of 1 mA failed to take effect (P > 0.05). After 3 and 6 times of treatments, both electroacupuncture of 2 mA and 1 mA had the effect of increasing the pain threshold (all P < 0.01), and the effect of 2 mA was superior to that of 1 mA (P < 0.05), had the synergistic effect with 5 mg/(kg x d) Morphine (P < 0.05). After 6 times of treatments, both electroacupuncture of 2 mA and 1 mA could inhibit the expression of GFAP (both P < 0.01), and there was no significant difference between them (P > 0.05). Both of 5 mg/(kg x d) and 2.5 mg/(kg x d) of Morphine, however, didn't bring about inhibition effect (P > 0.05).

CONCLUSIONThere is a does-effect relationship on electroacupuncture of different current intensity for alleviating the tibial cancer pain in rats. The electroacupuncture with 2 mA, which is better than that with 1 mA, has the synergistic effect with 5 mg/(kg x d) of Morphine. The electroacupuncture can inhibit the expression of GFAP to cooperate with Morphine for the purpose of alleviating the rats' tibial cancer pain.

Animals ; Bone Neoplasms ; complications ; genetics ; metabolism ; Electroacupuncture ; instrumentation ; methods ; Female ; Gene Expression Regulation, Neoplastic ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Humans ; Pain ; etiology ; genetics ; metabolism ; Pain Management ; instrumentation ; methods ; Rats ; Rats, Wistar ; Spine ; metabolism ; Tibia ; metabolism

Objective To investigate the absorption mechanism of Bio-chanin A ( BCA) and PEGylated BCA loaded nanostructured lipid carri-ers ( PEG -BCA -NLC ) across Caco -2 cell model.Methods Caco-2 cell monolayer model was used to explore the uptake , transport and impact factors of BCA and PEG -BCA-NLC.Drug concentration was measured by HPLC and the apparent permeability coefficients ( Papp ) were calculated.Results The main mechanism of BCA and PEG-BCA-NLC across Caco -2 monolayer model was passive diffu-sion.But under the same time , lower uptake of drug in Caco -2 cells was observed at PEG -BCA -NLC group comparing to that at BCA group.The absorption and the apparent permeability coefficient of BCA in Caco-2 cells monolayer were increased when combined with verapam-il.The Papp (AP→BL) increased and Papp (BL→PL) reduced, but the uptake content was much lower than BCA solution.Conclusion Ab-sorption process of PEG -BCA-NLC in Caco-2 cells monolayer met the characteristics of passive diffusion.However , the PEG-BCA-NLC may not significantly improve the absorption of BCA in intestinal epithelial.

Objective To investigate the expression of osteopontin and matrix metalloproteinase-9 (MMP9),and the relationship of osteopontin and MMP9 in malignant melanoma.Methods Expression of osteopontin and MMP9 was measured by immunohistochemical SP method in 23 patients with primary cuta- neous malignant melanoma,17 patients with metastatic melanoma and 20 patients with pigmented nevus. Results Osteopontin and MMP9 were expressed respectively in 87.5% and 75.0% of 40 malignant melanoma specimens,15.0% and 10.0% of 20 pigmented nevus specimens.The expression of both osteo- pontin and MMP9 was significantly higher (both P＜0.05) in malignant melanoma than in pigmented ne- vus.There was no correlation between the expression of osteopontin and MMP9,with age,sex,lymph node metastasis or location of lesions (P＞0.05).Twenty-nine cases were positive for both osteopontin and MMP9,4 negative for either osteopontin or MMP9.Conclusion Both osteopontin and MMP9 were over- expressed in malignant melanoma,but neither was related to lymph node metastasis.