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 study the isolation,culture, adipogenic and osteogenic induction Tupaia bone marrow mesenchymal stem cells(BM-MSCs).Method The BM-MSCs from tupaia were isolated and expended by combination of gradient centrifugation and adherence culture , then subcultured and observed for morphology under inverted phase contrast microscope.BM-MSCs were induced to adipocytes .and osteoblasts in vitro Result Cells were spindle or triangle-shaped, and clone proliferation .Cells were successfully induced into adipocytes .and osteoblasts Conclusions The method of isolation BM-MSCs from tupaia by combination of gradient centrifugation and adherence culture is simple and feasible , BM-MSCs have differentiation potential into adipocytes and osteoblasts .

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 .

Tree shrews get more and more concerns due to many of its physiological , biochemical and anatomical characteristics similar to those of human beings .Therefore, tree shrews models of human diseases such as viral diseases , neurological diseases and tumors attract more and more attention of researchers .In this article we will review the recent ad-vances in application of tree shrew models in research on human viral diseases .

Viral hepatitis is a major liver disease caused by virus infection .Viral hepatitis is popular in China , mainly caused by hepatitis B and hepatitis C viruses .Experimental animal model is a necessary platform for the research on mechanism of viral infection and pathogenicity , for treatment and vaccine development .Up to date, a great progress in the development of viral hepatitis animal models has been achieved in spite of the most of findings are limited to hepatitis B and C.Here, we summarize the recent findings of viral hepatitis animal models , focusing on the tree shrew animal model and its modeling strategy .

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 investigate the infection status of Toxoplasma gondii in different colonies of tree shrews and then provide the basis for parasitological monitoring .Methods Each of the forty blood samples were randomly collected from three tree shrews colonies : wild origin, domesticated and first generation, respectively.Both indirect hemagglutination test (IHA) and PCR assay were used to detect the Toxoplasma gondii.Results No positive sample of Toxoplasma gondii was detected from either IHA or PCR results .The results from IHA and PCR assays were in coincidence with each other.Conclusions According to the survey none of the tree shrews from the three groups is infected with Toxoplasma gondii.More samples or infection experiments are needed to determine whether tree shrews can be infected with Toxoplasma gondii.

Objective We established a rapid detection method of Pasteurella spp.and provided a reference for microbiological quality control of laboratory animal .Methods According to the β subunit of bacterial RNA polymerase ( rpoB) protein multiple alignments of 13 different Pasteurella spp.published in NCBI .The degenerate primers were designed by CODEHOP designer online .CODEHOP PCR method was applied to detecting Pasteurella spp.after the specificity and sensitivity of the method had been evaluated by 21 reference strains .Results Standard strain amplified fragment were about 200 bp by degenerate primers PastF6/PastR5.The primers are able to distinguish between Pasteurella spp.and the other pathognic organisms of laboratory animal respiratory tracts .Sensitivity of this method were 0.2 pg/μL~2 pg/μL to different Pasteurella.The Pasteurella positive rate was 19.1% in 609 animal ' s respiratory samples .The accuracy of positive results was 100%through verifying by sequenced and blast .Conclusions The established method has good specificity and sensitivity .It can be used to detect Pasteurella spp.in animal samples .

Objective:To study the effect of medical social workers combined with ICU diary on the prevention and treatment of post-traumatic stress disorder in ICU traffic accident patients, and to explore the application of continuing nursing services and ICU diary to ICU traffic accident patients.Methods:A total of 47 patients who were admitted to the ICU of Suzhou Science and Technology City Hospital from January to December 2019 due to traffic accidents were selected by convenient sampling method. Twenty-three patients admitted to the hospital in the first 6 months were selected as the control group (before the introduction of medical social workers and ICU diaries) for routine ICU treatment and nursing mode. Twenty-four patients admitted to the hospital in the last 6 months were selected as the experimental group (after the introduction of medical social workers and ICU diaries), and the medical social workers combined with the ICU diary were intervened. The scores of the Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS) were compared between the two groups of patients when they were admitted to the ICU, stayed in the ICU for a week, and transferred out of the ICU. The incidence of post-traumatic stress disorder was evaluated and compared by the scores of the Impact of Event Scale-Revised of the two groups of patients one month after leaving the ICU, three months after leaving the ICU, and six months after leaving the ICU.Results:The scores of SAS in the experimental group were 58.67 ± 7.45, 44.13 ± 5.47 and 39.08 ± 5.52 when they were admitted to the ICU, stayed in the ICU for a week, and transferred out of the ICU, and were 58.17 ± 6.75, 50.87 ± 6.35 and 42.74 ± 4.85, respectively in the control group. There was no statistical difference in the scores of the SAS between the two groups of patients when they were admitted to the ICU ( P>0.05), but there was a time difference between the two groups with time changes and different interventions ( Ftime = 110.98, P<0.01), there were also differences between groups ( Fbetween groups = 5.91, P<0.05). Compared with the control group, the SAS score of the experimental group decreased more significantly( Finteraction = 0.28, P<0.05). The scores of SDS in the experimental group were 57.75 ± 4.06, 45.29 ± 3.39 and 36.63 ± 3.49 when they were admitted to the ICU, stayed in the ICU for a week, and transferred out of the ICU, and were 57.48 ± 2.29, 49.04 ± 5.65 and 43.57 ± 4.07 respectively in the control group. There was no statistical difference in the scores of the SDS between the two groups of patients when they were admitted to the ICU ( P>0.05), but there was a time difference between the two groups of patients with time changes and different interventions ( Ftime = 248.24, P<0.01), there are also differences between groups ( Fbetween groups = 24.39, P<0.05). Compared with the control group, the SDS score of the experimental group decreased more significantly ( Finteraction=10.44, P<0.05). The incidences of post-traumatic stress disorder in the experimental group were 9, 8, 6 cases at one month, three months and six months after leaving the ICU, which were lower than 12, 11, 8 cases in the control group. The generalized estimation equation showed that the difference was statistically significant ( χ2=4.21, P<0.05). Conclusions:The intervention model of medical social workers combined with ICU diary is conducive to the use of ICU diary and the development of continuous nursing, which improves patients′negative emotions and reduces the incidence of post-traumatic stress disorder.