Objective To establish a tree shrew model of Fusarium solani keractitis by injecting Fusarium solani conidia into the corneal stroma.Methods Fusarium solani was inoculated into Sabouraud culture medium and incubated at 26℃ for 7 days.Fungal suspension was collected and the number of spores was adjusted to 1 × 1010 CFU /mL on the blood cell count plate.Forty healthy tree shrews were randomly divided into experimental group (n=30) and control group (n=10).In the experimental group, 50 μL of fungal spore suspension was injected into the cornea center with a 29G needle, and 50 μL saline was injected in the control group.The models were evaluated by anterior segment photography, in vivo confocal microscopy, histopathology, and corneal tissue culture.Results The fungal infiltration, the degree of edema of corneal epithelial and endothelial cells, and the number of mycelium were positively correlated with time.The number of infiltrating inflammatory cells, mainly, neutrophils, reached a peak on the 7th day after modeling.The mycelial growth was parallel to the stromal fibers.After the successful establishment of the model, the corneal tissue culture showed the growth of Fusarium solani.The successful rate of modeling was 86%.Conclusions The tree shrew model of Fusarium solani keratitis is established by injecting spores of Fusarium solani into the cornea.

Objective To analyze the imaging features of hepatic angiomyolipoma (HAML) on computed tomography (CT) and magnetic resonance imaging (MRI).Methods The imaging fingdings of 18 tumors which were pathologically diagnosed as HAML after surgery were analyzed retrospectively.Before operation,twelve and ten patients underwent CT and MRI non-contrast and dynamic enhanced scans,respectively,and 4 patients received both examinations.The imaging characteristics including the number,diameter,location,appearance of the lesions,plain and dynamic enhancement mode were analyzed.Results Eighteen HAML lesions were found in 18 patients with a diameter ranging from 1.5 cm to 17.2 cm (mean 5.3 cm).Five lesions manifested fatty content,and one showed hemorrhage and necrosis.Five HAMLs enhanced in a fast-in and fast-out mode,eleven in a fast-in and slow-out mode and two other lesions in an irregularly discrete mode.The arterial supply was found in 11 HAMLs in the hepatic arterial phase,all coming from intrahepatic arteries.Intratumoral vessels were observed in 12 HAMLs.Early draining veins to the hepatic vein (n =3) and portal vein (n =1) were detected in 4 HAMLs.One lesion demonstrated delayed enhancement in the pseudocapsule.Conclusion The detection of arterial supply and intratumoral vessels in a hypervascular hepatic tumor on contrast CT and MRI in a patient with a non-cirrhotic liver and normal AFP helps to make a diagnosis of HAML.

Objective To study the antagonistic effects of magnesium-selenium-fluorine preparation on dental fluorosis of mice and its mechanism,and to provide a foundational basis for prevention and control of dental fluorosis.Methods Eighty male SPF ICR mice,were divided into 8 groups according to body weight by random number table method:control group,magnesium group,selenium group,magnesium-selenium group,fluoride group,magnesium-fluorine group,selenium-fluorine group and magnesium-selenium-fluorine group.The control group,magnesium group,selenium group and magnesium-selenium group drank double steamed water,and the other four groups drank 50 mg/L F-double steamed water.The control group and fluoride group fed conventionally.Magnesium group and magnesium-fluorine group fed conventionally by adding MgSO4·7H2O 162.5 mg/kg.Selenium group andselenium-fluorine group fed conventionally by adding Na2SeO3 ·5H2O 2.0 mg/kg.Magnesium-selenium group and magnesium-selenium-fluorine group fed conventionally by adding MgSO4·7H2O 162.5 mg/kg and Na2SeO3·5H2O 2.0mg/kg.Then incisor specimens were obtained after the mice were put into death after treatment for 42 days.The expression of amelogenin was observed with immunohistochemical staining.And gray value is expressed as a result,the greater the gray value,the less protein expression.Results The results of light microscope showed that ameloblasts in the fluoride group showed disarrangement and even had vacuolar.The change of ameloblasts in the magnesium-selenium-fluorine group had no significant difference with control group.The expressions of amelogenin in enamel matrix of fluoride group (131.03 ± 11.14) was significantly higher than those of others (control group:143.44 ± 2.52,magnesium group:143.73 ± 12.43,selenium group:148.89 ± 2.85,magnesium-selenium group:148.38 ± 7.58,magnesium-fluorine group:145.90 ± 7.00,selenium-fluorine group:148.70 ± 4.90,and magnesiumselenium-fluorine group:151.89 4± 4.59,all P ＜ 0.05).The expression of amelogenin in ameloblast of fluoride group (165.49 ± 5.66) was significantly lower than those of control group,magnesium group,magnesium-fluorine group and selenium group (151.35 ± 2.52,149.27 ± 11.13,146.21 ± 4.84 and 150.39 ± 6.65,all P ＜ 0.05).The expression of amelogenin in ameloblast of selenium-fluorine group (165.46 ± 5.81) was significantly lower than those of magnesium group,magnesium-fluorine group and selenium group (all P ＜ 0.05).The results of factorial analysis showed that magnesium and selenium affected the expression of amelogenin in enamel matrix (F =4.195,15.009,all P ＜ 0.05),the interaction between fluoride and magnesium had an effect on the expression of amelogenin in enamel matrix (F =4.402,P ＜ 0.05).Interaction of fluoride,magnesium and selenium had no effect on the expression of amelogenin in enamel matrix (F =1.561,P ＞ 0.05).The fluoride,magnesium and selenium affected the expression of amelogenin in ameloblast (F =18.463,9.372,4.741,all P ＜ 0.05),the interactions between fluoride and magnesium,magnesium and selenium had effects on the expression of amelogenin in ameloblast (F =10.351,5.919,all P ＜ 0.05).Interaction of fluoride,magnesium and selenium had no effect on the expression of amelogenin in ameloblast (F =1.460,P ＞ 0.05).Conclusions There is an antagonistic effect of magnesium on the expression of enamel protein in fluorosis mice.However,it can not be considered that there is an effect of selenium on the expression of enamel protein in fluorosis mice.

Since 2010,the activities of high quality nursing service demonstration project were carried out in the national health system,its main purpose is to strengthen the nursing foundation,and provide satisfactory service.By conducting these activities,significant achievements have been made,such as improved nursing quality,reduced nurse-patient disputes.However,in the process of implementation,there are stillsome problems for us to think about and solve.In this paper,there is a brief commentary about the results of high quality nursing implementation and related issues.

Objective To isolate and identify viruses from fecal samples of tree shrew with diarrhea.Methods Fecal sample supernatant of tree shrew with diarrhea was inoculated to three cell lines ( Vero, LLC-MK2 and KMB17 ) , and the cytopathic effects on the cells were observed.The infectious particles in the culture supernatant were further ana-lyzed by transmission electron microscopy ( TEM) , genomic RNA-PAGE, rotavirus detection kit, amplification of S1 com-plete segment and bioinformatics analysis.Results Constant cytopathic effects were induced in Vero, LLC-MK2 and KMB17 cell lines after three passages of culture.The results from TEM, RNA-PAGE and rotavirus analysis indicated that they belong to reoviruses.Analysis of the S1 segments revealed that the S1 sequence from KMB17 cell culture had the high-est homology with that of prototype isolate T1L (85%nucleotide homology and 90%amino acid homology), therefore this isolate was named as type I reovirus.The other two S1 sequences from LLC-MK2 and Vero cell culture were identical to have 85%nucleotide homology and 92%amino acid homology with the prototype isolate T3D, named as type III reovirus. Phylogenetic analysis indicated that the isolates in this study are evolutionally adapted to tree shrews.Conclusions It is the first report here that 2 genotypes of Tupaia orthoreovirus are isolated and identified from one fecal sample via three cell lines and viral S1-specific primers, which provides useful guidelines for the isolation and identification of other reoviruses from tree shrew or other hosts.

Objective To screen out specific microsatellite markers for use in Tupaia belangeri chinensis genetic testing. Methods Firstly to screen about 700 microsatellite loci from whole genome.Secondly to choose about 100 better loci without defect factors.Lastly 46 primers were designed by 33 tree shrew’ s microsatellite loci obtained from whole genome and other references.Agarose gel electrophoresis and polyacrylamide gel electrophoresis were used for PCR products, and better loci based on electrophoresis results were chosen.Then STR scan was used to select the microsatellite loci combination for genetic testing.Results Twenty-two microsatellite loci were selected with a significant Stutter peak on STR scanning.Comparing the alternative loci and ultimately selected loci, there were two loci available in the five alternative loci of T.glis.The coincidence rate between T.glis and T.b.chinensis was 40%.There were two loci available in the five alternative loci of T.minor, and the coincidence rate between T.minor and T.b.chinensis was 40%.There were two loci available in the three alternative loci of T.belangeri, and the coincidence rate between T.belangeri and T.b. chinensis was about 70%.Conclusions The 22 microsatellite loci screened in this study are well applied for genetic testing of Tupaia belangeri chinensis, therefore, provide a scientific basis for the genetic quality monitoring of tree shrews.

Objective To establish a reverse transcription nested polymerase chain reaction ( RT-nPCR ) assay for detection of tree shrews orthoreovirus (TRV).Methods Three strains of TRV were respectively isolated from fresh feces of three tree shrews that came from the same field at different times .We designed and synthesized two pairs of MRV L1 gene nested primers and established the system of RT-nPCR.The TRV RNA was extracted and reversely transcribed to cDNA as a template for nested-PCR amplification.The developed RT-nPCR was optimized.The specificity and sensitivity were tested.Finally, the RT-nPCR was used to detect TRV in 25 tree shrew samples.Results Taking the genomic RNA of TRV as template, the RT-nPCR was able to amplify a specific fragment band targeting the L 1 gene, while there were no target bands in the normal cell control , ( Wa strain rotavirus , hepatitis A virus , and herpes simplex virus ) .The RNA of TRV was diluted by 1:10 to 1:109 .Each dilution sample was analyzed by the RT-nPCR.The minimum detectable concentration of RNA was 0.01 pg/μL.The results of RT-nPCR detection showed that 4 of the 15 tree shrews were TRV-positive in the survival group , and 10 of 10 tree shrews were TRV-positive in the death group . Conclusions The RT-nRCR assay established in this study is accurate , specific and sensitive .Therefore, it can be used for routine detection of TRV in quality assurance testing .

Objective To understand the histological characteristics of the major endocrine organs of tree shrew , and provide a normal histological atlas of endocrine organs of tree shrew .Methods Ten artificially fed healthy tree shrews were killed and dissected after anesthesia .The thyroid, parathyroid, adrenal and pituitary glands were observed by gross inspection and samples were taken for routine histological examination with HE staining .Results ( 1 ) The thyroid gland was pale yellow, located on both sides of the 2-4 tracheal rings.The thyroid gland was plate-shaped, its surface was covered with a thin fibrous capsule . The thyroid parenchyma was divided into several lobules by stretched capsule membrane .Follicular and parafollicular cells were distributed in the lobules , and red colloid was present in follicular cavity.(2) Each side had one parathyroid , located on the cranial or the outer surface of the middle part of the thyroid gland, and was slightly covered by thyroid .The gland was round or oval , and its parenchyma was made up of the principal cells and eosinophil cells , and acinar structure appeared in the parenchyma .( 3 ) The adrenal glands were oval , yellow color, located in the renal hili , and linked to the kidneys .They were surrounded by a thin capsule .The parenchyma was divided into cortex and medulla .The cortex was divided into zona glomerulosa , zona fasciculata and zona reticularis from outside to inside.The zona glomerulosa was the thickest layer and the zona fasciculata was the thinnest .The medulla cells formed clumps or mesh, with central vein in the central part .(4) The pituitary gland was located in the sella turcica , with no recessus hypophysis .The pituitary gland was composed of the adenohypophysis and neurohypophysis .Its surface was covered with a connective tissue capsule .The pituitary gland was divided into distal part , middle part and pars tuberalis . neurohypophysis was made up of neural and pars infundibularis .Conclusions The histological atlas of endocrine organs in the tree shrew is established , which is close to that of the primate animals in the morphology , and provide histological evidence for the study of tree shrew endocrine organs and disorders , as well as the animal model of human diseases .

BACKGROUND:In recent years a large number of studies have suggested that bone marrow mesenchymal stem cells can ease hyperglycemia of diabetic rats, but the related mechanism is unclear and controversial. OBJECTIVE:To investigate the relevant mechanism of bone marrow mesenchymal stem cells on pancreas microenvironment in vivo in diabetic rats. METHODS:Bone marrow mesenchymal stem cells were transfected with enhanced green fluorescent protein (EGFP) and administered to diabetic rats via the subcapsular pancreas. Blood glucose levels were monitored. The expressions of the key genes in islet development in these EGFP positive pancreatic cells were analyzed by Real-time quantitative PCR at different times. EGFP and insulin double-positive cells were detected by immunofluorescence. Flow cytometry was performed to analyze cellcycle and DNA ploidy. RESULTS AND CONCLUSION:Blood glucose levels were effectively reduced after transplantation. The expressions of the key genes in islet development reached their own peak values at different times after transplantation:Nestin at week 1, Nkx 2.2 at week 3, Pax 4 and Ngn 3 at week 4, insulin and glucagon at week 12, PDX-1 at week 8 until week 12. The cells double-positive for EGFP and insulin cells were observed. In the pancreas, EGFP positive cells at S+G 2/M phase were significantly increased, and there were no polyploid and aneuploid cells. In pancreas microenvironment, the bone marrow mesenchymal stem cells transplanted into the diabetic pancreas can differentiate into isletβ-like cells under gene control, but not through the fusion with tissue cells.

Objective To investigate the expression of glucose transporter type 1(GLUT-1)and hypoxia-inducible factor 1 (HIF-1)α in psoriatic lesions,and to explore their correlations with keratinocyte proliferation.Methods Biopsy specimens were obtained from 30 patients with psoriasis and 20 normal human controls.Immunohistochemistry and Western blotting were used to examine the protein expression of GLUT-1 and HIF-1α in these specimens.Results GLUT-1 and HIF-1α were mainly expressed in the basal layer of the control skin,but throughout the whole epidermis of psoriatic lesions.A significant increase was observed in the expression of GLUT-1 and HIF-1α in psoriatic lesions compared with that in the control skin (botb P＜0.01).In the case of psoriatic lesions,both the expression of GLUT-1 and HIF-1α was positively correlated with that of Ki-67(r=0.70,0.81 respectively,both P＜0.01),and positive correlation was also found between the expression of GLUT-1 and HIF-1α(r=0.85.P＜0.01).Conclusion Our data suggest that uprcgulation Of GLUT-1 and HIF-1α expression in psoriatic lesions might contribute to the proliferation of keratinocytes and psoriasis development.