Objective To investigate the application of multi-slice spiral computed tomography(MSCT) in the diagnosis of solid pseudopapollary tumor of the pancreas (SPTP). Methods Clinical data and CT films of 12 patients with SPTP were retrospectively analyzed from January 2003 to December 2008. Results SPTP presented a typical cystic lesion with well-limited silhouette and no intensification in the cyst on enhanced CT scan. However, a slight to moderate enhancement in the solid components and a markedly enhanced envelope could be seen. Three dimensional images of MSCT can reveal clearly an anatomic relationship of the lesions with surrounding organs and blood vessels. Of 12 cases, there was one case showed that the envelope was incomplete, 3 with duodenal invasion, 2 with superior mesenteric vein involvement, and 1 with closed adhesion with spleen. All 12 patients underwent surgery and had only one tumor, tumor diameter ranged from 4 cm to 18 cm. The location of tumor in pancreas, the relation with surrounding tissue and the pathological presentation were helpful to make peroperative diagnosis. Three dimensional imaging technology of MSCT can offer important referrence for the preroperative evaluation and increase the diagnosis accuracy.

Objective To investigate the morphological changes of bone marrow megakaryocytes in patients with bacterial and fungal infection.Methods Totally 76 patients with microorganism infection from the Second Affiliated Hospital,Zhejiang University School of Medicine from January 2008 to August 2009 were enrolled,including 56 bacteria infected patients and 20 fungal infected patients.All patients received bone marrow examinations,and were positive in microorganism culture.Thirty subjects without infection,hematological disease and other severe diseases were randomly selected as controls.The number and function of megakaryocytes were examined retrospectively, and the size, nuclear lobulation, and vacuolar degeneration of megakaryocytes were quantitative analyzed and compared among the groups.Results The size,nuclear lobulation,vacuolar degeneration,and Yat nuclear of megakaryocytes in bacterial infected group were 2.20 ±0.21,2.11 ±0.23,0.51 ±0.11 and 0.74 ±0.11 respectively,those in fungal infected group were 2.21 ±0.16,2.10 ±0.19,0.52 ±0.10 and 0.79 ±0.10 respectively;while those in control group were 1.40 ±0.10,1.36 ±0.12,0.28 ±0.06 and 0.54 ±0.09 respectively.The differences between bacterial infected group and control were of statistical significance(t values were 14.52,12.19,9.33 and 6.61 respectively,P ＜ 0.05),and the differences between fungal infected group and control were of statistical significance(t values were 16.27,12.34,7.85 and 6.49 respectively,P ＜ 0.05).The size,nuclear lobulation,and vacuoles of megakaryocytes in gram-negative(G-)bacteria group were 2.29 ±0.20,2.22 ±0.26 and 0.57 ±0.10,while those in the gram-positive(G+)bacteria group were 2.13 ±0.20,2.04 ±0.18 and 0.46 ±0.09,and the differences were also significant(t values were 2.07,3.03and 3.56 respectively,P ＜ 0.05).The production of platelet by megakaryocytes in bacterial infected group,in fungal infected and the control were 31.4 ±7.6,32.4 ±6.4 and 41.3 ±5.5,and the differences between bacterial infected group and control,fungal infected group and control were significant(t values were 4.78and 3.98 respectively,P ＜ 0.05).The production of platelet in G-bacteria group was 28.0 ± 6.7,while that in G + bacteria group was 34.4 ± 7.2,and the difference was also of statistical significance(t = 2.41,P ＜0.05). Conclusion Bacterial infected patients have increased megakaryocytes cell body,nuclear lobulation,obvious vacuolar degeneration,Yat nuclear and decreased platelet production function,which are more significant in G- bacteria infected group.

BACKGROUND:As current studies on isolation, culture andcryopreservation of human amniotic epithelial cells (hAECs) are relatively scattered, it is difficult to form a comprehensive and effective solution to meet the clinical needs of stem cells for transplantation in future.OBJECTIVE:To establish the technology of isolation, culture and cryopreservation of hAECs, and to study the biological characteristics of hAECs.METHODS:Orthogonal method was used to study the effects of different factors on the separation, culture and cryopreservation, and range method was adopted to analyze the data to optimize the separation, culture and cryopreservation. We performed cell primary and passage cultures, morphology observed by microscope, drawn cell growth curve and flow cytometry assay, immunofluorescence staining, hepatocyte like cell differentiation to study the biological characteristics of hAECs.RESULTS AND CONCLUSION:(1) The optimal hAECs separation conditions were as follows:trypsin digestions were conducted at a concentration of 0.25%, four times, once for 20 minutes digestion; optimal conditions of culture were 4×108/L cell seeding density, 10 μg/L epidermal growth factor, 5% serum; optimal conditions of cryopreservation were 1×1010/L cell cryopreservation density, 10% dimethyl sulfoxide, 80% serum. (2) The primary cells were adhered to the wall in 2-3 days, exhibiting irregular polygon, paving stone-like growth. Cell adherence and growth rate were accelerated after subculture, and the growth and proliferation ability of passage 2 cells were not significantly decreased after cryopreservation and resuscitation. (3) Immunofluorescence staining showed that the primary cells strongly expressed SSEA-4 and CK19, but did not express Vimentin, CD45 and HLA-DR. The immunophenotype statistics of the primary and passage 4 cells showed the epithelial mesenchymal transition of hAECs in culture process. (4) Immunofluorescence staining showed that the liver cell marker expression of ALB, CK18 was significantly increased after hAECs were induced to differentiate into hepatocyte-like cells. Glycogen staining revealed glycogen synthesis in hAECs after 3 weeks of induction. To conclude, hAECs are easy to obtain and have strong proliferation ability in vitro, and express surface markers for undifferentiated embryonic stem cells.

Objective To investigate the role of inflammatory cytokines in the pathogenesis of chronic non-bacterial prostatitis/chronic pelvic pain syndrome (CAP/CPPS) patients. Methods The 38 cases with CAP/CPPS patients (18 cases of CAP and 20 cases of CPPS) and 20 cases of healthy controls were selected. The differential expressions of 40 kinds of inflammatory cytokines were detec-ted by antibody arrays in prostate fluid. Results The inflammatory cytokines which increased more than 1.5 times expression have been found. There were seven kinds in CAP including monocyte che-moattractant protein (MCP)-1, solution tumor necrosis factor receptor Ⅱ(s TNF R Ⅱ), platelet-de-rived growth faetor-BB (PDGF-BB), interleukin (IL)-β, IL-11、IL-6、MCP-2 and five kinds in CPPS groups including MCP-1、PDGF-BB、MCP-2、s TNF R Ⅱ、It-11 respectively, compared with healthy control group. The cluster analysis results showed that protein expression of Monocyte chemoattrac-tant protein 1 (MCP-1)and platelet-derived growth factor BB (PDGF-BB) were significantly increased in CAP (3.47 and 2.07 times) and CPPS (2.25 and 2.19 times) compared with healthy control group and were the final polymerization of inflammatory cytokines. The protein expression of interleukin 1 β (IL-1 β), MCP-1 and soluble tumor necrosis factor Ⅱ (s TNF R Ⅱ) in CAP group was increased more than 1.85,1.55,1.67 times compared with CPPS group. Conclusions Elevated expression of inflammatory cytokines may play an important role in the course of CAP/CPPS disease. The extent of the inflammatory response of CAP was higher than CPPS. The inflammatory factors of MCP-1 and PDGF-BB could serve as a novel diagnostic marker.

Objective To compare the dosimetric difference between RapidArc and fixed gantry angle dynamic IMRT (dIMRT) for central-type lung cancer radiotherapy. Methods Therapy for 10 patients previously treated with dIMRT was replanned with RapidArc. Dose prescription was 66 Gy/33 fraction. Comparative endpoints were planning target volume (PTV) dose, doses to surrounding structures,number of monitor units, and treatment delivery time. Results There was no significant dosimetric difference between RapidArc and dIMRT. Compared with dIMRT, RapidArc slightly elevated target volume dose, lung V5, V10. The average values of lung V20, V30 and heart V30 were larger in dIMRT than those in RapidArc. The number of monitor units was reduced by 32% and the treatment time by 66% in RapidArc.Conclusions Both RapidArc and dIMRT plans could meet the clinical therapy needs. RapidArc could achieve similar target coverage and sparing of organs at risk, with fewer monitor units and shorter delivery time than dIMRT.

Objective To evaluate the effectiveness of single balloon enteroscopy (SBE) in diagno-sing of suspected lesions in small intestine. Methods Data of 23 patients with suspected small intestinal disease, who underwent SBE (Olympus) between February 2009 and August 2009, were retrospectively studied. A total of 34 procedures were performed in 23 patients. The indications for the examination were suspected obscure gastrointestinal bleeding (n = 9), abdominal pain (n = 7), suspected intestinal tumor re-vealed by capsule endoscopy (n = 4), and Crohn disease (n = 3). Results The average preparation time of SBE was less than 5 minutes. The mean procedure time was 61±25 minutes and 67±28 minutes for the oral and anal routes, respectively. Examination of whole length of small intestine was achieved in 6 patients. The diagnostic rate of small-intestinal lesions was 60. 9%, and no severe complications including perforation occurred. Conclusion SBE is safe and easy to prepare and perform, which can be a useful diagnostic and therapeutic tool for suspected small bowel disease.

Objective To express the hemagglutinin of H7N9 in Pichia pastoris and analyze its immunogenicity. Methods The HA [1 -525 amino acids(aa)] of H7N9 [A/Hangzhou/1/2013(H7N9)] lacking the C-terminal transmembrane anchor-coding sequence was amplified by PCR and cloned into the expression vector pPICZαA.The plasmid pPICZ-HA7-S-was transformed into P.pastoris and the HA1-525 was detected by ELISA.Then the HA1-525 was precipitated by PEG20000.After immunization with the HA1-525 , the anti-HA7 antibody in mouse serum was detected by ELISA and hemagglutinin inhibition ( HI) test.Results The HA1-525 was expressed in P.pastoris after being induced with methanol. Western blotting confirmed that there were specific dispersion bands and the molecular weight of HA1-525 decreased to 58 × 103 after being digested by endo H.Anti-HA7 antibody was found in serum of HA1-525 immunized mice and the hemaggluti-nation-inhibition titers reached 1∶700 after the third doses.Conclusion This study shows the HA1-525 expressed in P.pastoris can induce the neutralizing antibody in mice.

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

Objective:Immunoregulation study of umbilical mesenchymal stem cell (UCMSCs) on allogeneic umbilical cord blood(UCB) CD4+T lymphocytes,which proliferation,apoptosis and the differentiation to CD4+CD25+ regulatory T cell (Treg) in vitro. Methods:Establishing on direct contact or transwell co-culture system,adopt in different proportion of UCMCs with phytohaemag-glutinin (PHA)-activated UCB CD4+T lymphocytes were co-cultured. The proliferation of lymphocyte,percent of CD4+CD25+/CD4+and Foxp3 expression, regulatory T cell marker gene were measured. Apoptosis of CD4+T lymphocytes were observed in the direct contact or transwell coculture system of UCMSCs with desamethason( DXM)-stimulated UCB CD4+T lymphocytes. Results: The UCB CD4+T lymphocytes cocultured with UCMSCs with PHA-activating for 3 days,compared with the UCMSCs free control group,the amount of cells was reduced noticeably(P<0. 05) and the percent of CD4+CD25+in CD4+T lymphocytes and Foxp3 expression significantly in-creased(P<0. 01) in a dose dependent way(P<0. 05). The UCB CD4+T lymphocytes cocultured with UCMSCs with DXM-inducing for 7 days,the apoptosis rate was significantly lower than that of the control group without UCMSCs (P<0. 01). These effects were partially attenuated in transwell coculture but could not be eliminated. Conclusion: UCMSCs are negative effect on UCB CD4+T lymphocytes-mediated immunity effects,and mainly manifested in the regulation on cell proliferate ability and differentiation rather than promoting apoptosis.