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.

Purpose To compare radiation dose and image quality of different scan modes for CT pulmonary angiography (CTPA) including high-pitch flash mode, dual energy CT (DECT) mode, 128-slice mode of dual source CT and 64-slice CT mode. Materials and Methods One hundred and sixty-seven patients with suspected pulmonary embolism were retrospectively studied. All the patients underwent CTPA. Twenty patients were selected randomly from the patients scanned by high-pitch flash mode. Patients who were scanned by the other three modes were selected with body mass index and age matched those in high-pitch flash mode, with 20 patients in each group. Two radiologists assessed signal-to-noise ratio (SNR) and image quality with 5-piont scale. Dose parameters of volume CT dose index (CTDIvol), dose length product (DLP), and effective dose (ED) were compared among the four groups. Results Mean CTDIvol, DLP and ED were (3.72±0.74) mGy, (137.5±28.7) mGy · cm, and (2.34±0.41) mSv for Flash mode;(5.31±1.21) mGy, (181.6±34.5) mGy· cm and (3.24±0.57) mSv for DECT mode;(5.66±1.47) mGy, (198.7±42.1) mGy·cm and (3.58±0.63) mSv for 128-CT mode;and (6.75±1.68) mGy, (231.5±54.2) mGy·cm and (4.21±0.89) mSv for 64-CT mode. There was no significant difference of SNR and image quality among the four modes (P>0.05). Conclusion There are no significant difference of image quality among the four groups. Flash mode allows for minimum radiation dose compared to other modes. DECT mode and 128-CT mode get higher radiation dose with no difference between them. 64-CT mode gets the highest radiation dose.

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 .

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 .

Objective To explore the risk factors for upper gastrointestinal haemorrhage (UGH) in hepatocellular carcinoma (HCC) with portal hypertension (PH). Methods We retrospectively reviewed the medical records of 231 patients with HCC-PH treated in our Department from 1st January 2005 to 1st August 2009. The clinicopathologic factors were evaluated for their possible association with UGH in univariate analysis followed by multivariate analysis using Logistic regression model. The overall survival (OS) was calculated by the Kaplan-Meier method. Receiver operating characteristics (ROC) analysis with calculation of the area under the curve (AUC), sensitivity, and specificity were carried out to assess the predictive ability of the independent risk factors. Results Among 247 patients diagnosed with HCC-PH, 231 patients met the inclusion criteria and were entered into this study. UGH occurred in 28 patients (12.12 %, 28/231). Patients suffering from UGH had a higher 30-and 60-d mortality when compared with the non UGH group (53.57% vs. 4.43%, 96.43%vs. 10.34%, P＜0. 001, 0. 001). The 1-,2-and 3-year overall survival (OS) rates in the non-UGH and the UGH groups were 3. 57% (1/28), 0% (0/28), 0% (0/28) and 21.18% (43/203), 14.29% (29/203), 4.43% (9/203), respectively. There was a trend towards a non-significantly statistical difference in long-term (≥3 yr) survival (P=0. 605). UGH had a dismal prognosis with a median OS of 0. 8 months (0. 10-2. 40 months). Multivariate analysis of the risk factors showed elevated alpha-fetoprotein (AFP) (P = 0. 026) and aspartate aminotransferase (AST) more than twice normal (2N)(P=0. 004) were predictive factors, in particular, AST≥2N. A cutoff value (PI≥7. 242) predicted UGH with an AUC of 0.828 (95%CI, 0.698-0.957), sensitivity of 81.0% and a specificity of 81.0%, as calculated from the ROC. Risk score stratification predicted UGH to show a statistically significant difference (P＜0. 001). Conclusions UGH, as one of the end-stage incidents of HCC-PH,had a dismal prognosis. Patients with elevated AFP levels and AST levels above 2N were associated with high risks for UGH and should be monitored carefully or offered prophylactic treatments. Risk score stratification was useful for prediction of UGH.

Objective To evaluate factors affecting early recurrence and overall survival after curative hepatectomy for hepatocellular carcinoma (HCC) in cirrhotic patients. Methods Sixty two HCC cases with concomitant liver cirrhosis were retrospectively reviewed after curative hepatectomy in our department during the period between Jan. 2002 and Jan. 2009. Clinicopathologic data were evaluated for their possible association with postoperative early recurrence (ER) and overall survival (OS) in univariate analysis followed by multivariate analysis using COX proportional hazard model. Receiver operating characteristics (ROC) analysis with calculation of the area under the curve (AUC), sensitivity, and specificity was applied to assess predictive ability of independent risk factors. Results During follow-up period, 47 patients developed postoperative ER. The 1-, 2-, 3-and 5-year cumulative recurrence rate was 62.9% (39/62) ,75.8% (47/62), 80.7% (50/62), 83.9% (53/62) ,respectively. The 1-, 3- and 5-year OS rates were 59. 7% (37/62), 21.0% (13/62) and 1.6% (1/62), respectively. Multivariate analysis revealed that increased BCLC staging, severity of liver cirrhosis and tumoral residue resectional edge as independent risk factors influencing ER. Cutoff point value ( PI ≥2.171 ) predicted ER with AUC of 0.874(95%CI, 0.757～0.990), sensitivity was 85. 1% and specificity was 77. 8% calculated from ROC.Difference of median recurrence time according to risk stratification reached statistical significance ( 18.7mons, 7.7 mons vs. 2.9 mons, Log-rank test,λ2 =25. 288, P =0.000. While ER, post-recurrence treatment and severity of liver cirrhosis affected OS, cutoff point value ( PI ≥ 2. 893 ) predicted OS with sensitivity 86. 8%, and specificity 88.9%, with AUC 0.894(95% CI,0.798 ～ 0.990). Median OS time according to risk stratification demonstrated significant difference (27.8 mons, 21.5 mons vs. 8.5 mons,Log-rank test, λ2 = 30. 869, P = 0. 000). Conclusion Severity of liver cirrhosis and surgical tumor margin determines postoperative ER and OS for HCC after curative hepatectomy. Effective management of ER also contributes to good prognosis. Risk stratification can be used for evaluation of ER and OS of HCC.

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 .

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 determine the effectiveness and safety of stereotactic body radiotherapy (SBRT)-CyberKnife for oligometastatic prostate cancer.Methods From May 2012 to February 2017,31 patients treated by CyberKnife were retrospectively reviewed,with a median age of 67 years(range 52 to 83 years),including 50 oligometastatic and 2 primary prostate cancer patients.The median PSA level was 8.4 ng/ml(range 0 to 300.0 ng/ml) and PSA test was performed every month.PSA progression-free survival (PSA-PFS),time to initiation of androgen deprivation therapy (ADT) and local control rate (LCR) were measured as the main outcomes.Results SBRT was well tolerated and were performed as planned in all patients.No SBRT related acute or late toxicities were observed.No bone fracture was observed in patients treated by bony targeted radiotherapy.The median follow-up after SBRT was 20.7.months (range 1.2-58.3 months).The median PSA-PFS was 5.3 months (range 0-58.3 months).1-year,2-year,and 4-year PSA-PFS was 52.0％,36.7％ and 36.7％ respe ctively.PSA level decrease was observed in 21 oligometastatic prostate cancer patients after SBRT,with median PSA-PFS of 12.3 months (range 1.2-58.3 months).PSA level increase was observed in 29 oligometastatic prostate cancer patients after SBRT.Six local recurrence were observed resulting in an actuarial 1-year,2-year and 3-year LCR of 90.4％,86.9％ and 82.6％,respectively.Twelve patients treated without ADT after SBRT,with median follow-up of 8.6 months (range 2.9-58.3 months) in this subgroup.Seven patients were added ADT after SBRT,with the median time from SBRT to initiation of ADT of 13.3 months (range 3.0-24.0 months) in this subgroup.Twelve patients were treated with ADT continuously after SBRT.Conclusions CyberKnife seems to be a safe and effective treatment with tolerated adverse events and good local control for patients with oligometastatic prostate cancer.