Objective To analyze and study the relationship between plasma adhesion molecules,free amino acids and ovarian cancer.Methods A total of 67 patients with ovarian cancer in our hospital during the time of March 2015 to May 2016 were selected as the observation group,and 67 healthy women at the same time were selected as the control group.The plasma adhesion molecules and free amino acids levels of two groups were detected and compared,The detection levels of observation group with different stages and degree of differentiation of ovarian cancers were compared.The relationship between plasma adhesion molecules,free amino acids and ovarian cancer were analyzed by the Logistic analysis.Results The plasma adhesion molecules levels of observation group were all higher than those of control group (P ＜ 0.05),the plasma free amino acids levels were all lower than those of control group (P ＜ 0.05),and the detection levels of observation group with different stages and degree of differentiation of ovarian cancer plasma adhesion molecules and free amino acid levels had significant differences (P ＜ 0.05).The Logistic analysis showed that the plasma adhesion molecules and free amino acids had close relationship to the ovarian cancer (all P ＜ 0.05).Conclusions The plasma adhesion molecules and free amino acids of the patients with ovarian cancer show abnormal expression state,and the expression levels of patients with different stages and degree of differentiation of ovarian cancer have certain differences,so the detection value of those indexes in the patients with ovarian cancer is higher.

Objective To investigate the pathogenesis,high risk factors,clinical characteristics,methods of diagnosis and treatment,and prognosis of vaginal intraepithelial neoplasia (VAIN).Methods The clinical data of thirteen cases of VAIN treated in Zhejiang Provincial Cancer Hospital dated Mar.2002 through Dec.2008 were reviewed and analyzed retrospectively.Results Twelve of 13 VAIN cases were performed the human papillomavirus(HPV) detection with 92% (11/12) HPV positive rate.None of the cases shown specific clinical manifestation.Among the 13 cases,6 of them accompanied with cervical cancer,4 cases with cervical intraepithelial neoplasia (CIN ),and 3 cases with vulvar intraepithelial neoplasma (VIN).Five cases synchronously diagnosed with cervical lesion and 3 with vulva lesion were underwent surgery,while the other 5 cases were diagnosed metachronously.Among 8 cases underwent surgery,1 case with CIN underwent argon plasma coagulation (APC) after surgery,1 case with the positive edge of VIN underwent APC.During follow up,1 case with locally advanced cervical cancer underwent radiotherapy again,3 cases with VAIN received APC,while 1 cervical cancer cases with VAIN received no treatment.The average follow-up time was 25.6 months (range 6-87 months).Two cases died of cervical cancer metastasis.The other 11 cases were normal and still alive.None of them progressed to invasive carcinoma.Conclusions The main reason of VAIN is HPV infection.There are not specific clinical manifestations,usually diagnosed when reviewing cervical or vulva lesions and rarely progressed to invasive carcinoma.The main treatment of VAIN is surgery with the adjuvant treatment of APC.

Objective To study the sonographic features of mammary sclerosing adenosis (SA) and evaluate the diagnositic value of ultrasound. Methods Thirty-ifve patients with pathologically conifrmed SA in Ruijin Hospital from May 2009 to August 2013 were retrospectively analyzed. The Breast Imaging Reporting and Data System (BI-RADS) lexicon was introduced to describe the lesions. The diagnostic sensitivity was analyzed. Results The main sonographic ifndings of SA could be characterized into three types:(1) malignancy-looking nodule type (typeⅠ) (34%, 12/35). (2) benignity-looking nodule type (typeⅡ) (43%, 5/35). (3) architectural disorder type (typeⅢ) (23%, 8/35). Lack of blood supply has the greatest value in differential diagnosis among all sonographic features. The diagnositic sensitivity in typeⅠ, typeⅡ, typeⅢwere 0 (0/12), 93%(14/15), and 75%(6/8) respectively. The general sensitivity was only 57%(20/35). Conclusions There are no typical sonographic features in mammary sclerosing adenosis. Ultrasound doctors should improve their knowledge about this disease.

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 an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.

Objective To modify recombinant plasmid construction of BPO trimer,and establish the detec-tion of M2 type anti-mitochondrial antibody (AMA-M2) using ELISA.To modify the recombinant expression vector of the target gene triplet of AMA-M2,and to establish and evaluate an enzyme linked immunosorbent assay (ELISA) for the detection of AMA-M2.Methods The expression vector pGEX 4T 1 BPO was con-structed through the gene amplification of triplet BPO by polymerase chain reaction (PCR),and expressed in E.coli BL21,and the recombinant protein was induced by isopropyl Thioglucoside (IPTG).The target protein was purified by affinity chromatography,and ELISA was established to detect AMA-M2.Results The recom-binant protein with a purity greater than 95% was obtained.326 cases of clinical samples was examined,and in 43 cases of primary biliary cirrhosis (PBC),41 cases showed AMA-M2 positive,the sensitivity was 95.3%(41/43),the specificity was 98.2%(278/283),and the difference of AMA-M2 detection rate between PBC group and non PBC group was statistically significant.Conclusion The target protein obtained by the im-proved technology can be used for AMA-M2 detection.AMA-M2 has high sensitivity and specificity for the clinical diagnosis of PBC.

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 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 investigate the effects of recombinant adeno-associated virus-mediated myostatin propeptide (MPRO) on uptake and oxidation of glucose,and glycogen synthesis in C2C12 myotubes,as well as the associated molecular mechanism.Methods Mature C2C12 myotubes were assigned to the following 6 groups:control,insulin,green fluorescent protein (GFP),insulin + GFP,MPRO,and insulin + MPRO groups.Glucose uptake,glucose oxidation,and glycogen synthesis were detected by counting radioactivity of 14CO2 or 14C labeled glycogen derived from 2-deoxy-[1-14 C] glucose.The activity of insulin signal pathway was evaluated by Western blot.Results Compared with control group,glucose uptake and glycogen synthesis were significantly increased in insulin and insulin+GFP groups,and further increased in insulin+MPRO group as compared with insulin alone(all P＜ O.05).However,MPRO and insulin had no effect on glucose oxidation.The phosphorylations of insulin receptor (IR) β,insulin receptor substrate 1 (IRS-1),protein kinase B (Akt),glycogen synthase kinase-3 β (GSK-3β),and the expressions of phosphatidylinositol 3-kinase (PI3K) and glucose transporter 4 (Glut4) in membrane were significantly increased in insulin and insulin+GFP groups compared with control group(all P＜0.05),and were further increased after MPRO transfection (all P ＜ 0.05).Conclusion MPRO may increase insulin-stimulated glucose uptake and glycogen synthesis in C2C12 cells by activating the IRS/PI3K/Akt signal pathway.

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.