0.05) and decrease to higher than normal in CH and in 4 cases of SH (P

Objective To investigate the value of preoperative total abdominal spiral CT scan in evaluating the staging of ovarian cancer.Methods Clinical data of 42 patients with ovarian cancer undergoing total abdominal spiral CT were collected.The CT images were retrospectively analyzed,and the CT staging and surgical pathologic staging were compared.Results Multi-slice spiral CT on ovarian cancer invasion and metastasis has high accuracy of diagnosis.The diagnostic accuracy of this group for direct tumor invasion,peritoneal metastasis,lymph node metastasis and ascites was 66％-100％.The correct preoperative staging were 34 cases:6 cases in stage I,6 cases in stage Ⅱ,17 cases in stage Ⅲ,5 cases in stage Ⅳ,and the staging accuracy of 80.9％.Conclusion Total abdominal multi-slice spiral CT examination is of great value in preoperative staging of ovarian cancer.

Combining with advances in optogenetics and feedback control of physiological function, we have utilized self-made PPDP (preview, presentation, demonstration, promotion) teaching method to clarify how various physiological functions are regulated by the nervous system and carried out physiological innovation experiment activities. The innovative experiments aim to cultivate students' self-study capability, broaden their vision, enhance their interest in physiology, and finally promote the effect of physiological theory teaching. We herein summarize our practice of closed-loop control of innovative experimental teaching in optogenetics from the following four facets: education concept, students and teacher resources, teaching design, and teaching experience. This summary is trying to explore new experiences of promoting students' participation in teaching activities and improving the teaching quality of physiology.

Objective To investigate the effects of different cell cycles and their regulating genes on apoptosis of mononuclear-macrophages induced by Leptospira interrogans. Methods The diversity and alteration of cell cycles of murine mononuclear-macrophage line(J774A. 1 ) and human monocyte line(THP-1 ) before and after infected with L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai were detected using Cell Cycle Stain Kit plus flow cytometer. The cell cycle synchronized J774A. 1 and THP-1 cells were generated and then identified by using different cell cycle blocking agents and flow cytometer. By using Annexin V/PI Detection Kit combined with flow cytometer, the rates of early-apoptosis and late-apoptosis/necrosis in the synchronized and non-synchronized J774A. 1 and THP-1 cells after infection with L. interrogans strain Lai were determined. Several real-time fluorescence quantitative RT-PCRs were performed to the changes of mRNAs levels of p21, p27, p53, c-myc and cycA genes that associated with cell cycle and apoptosis in J774A. 1 and THP-1 cells before and after infected with L. interrogans strain Lai. Results There were G1, S and G2/M phases in both the non-infected normal J774A. 1 and THP-1 cells. On the contrast,the majority of infected J774A. 1 and THP-1 cells were stagnated at G1 phase, but the amount of S phase THP-1 cells was elevated while that of S phase J774A. 1 cells was not(P ＜0.05). No remarkable early-apoptosis in both the infected G1 phase J774A. 1 and THP-1 cells was found, whereas the rates of early-apoptosis and late-apoptosis/necrosis in the infected M phase J774A. 1 and THP-1 cells were significantly increased (P ＜0.05 ). Additionally, late-apoptosis / necrosis rate in the infected G1 phase THP-1 cells (P ＜ 0.05 )that not found in the infected G1 phase J774A. 1 cells. Compared to the non-infected cells, the p21 mRNA levels in the infected J774A. 1 and THP-1 cells were significantly elevated(P ＜0.05), and the c-myc and p27 mRNA levels in the infected J774A. 1 cells and the cycA mRNA level in the infected THP-1 cells were also higher than those in both the non-infected cells ( P ＜ 0.05 ). Conclusion Different cell cycles and their regulating genes have a role to affect the apoptosis of human and murine mononuclear-macrophages caused by L. interrogans with a diversity of cell line origins.

Objective To construct a plasmid DNA encoding G glycoprotein of respiratory syncytial virus(RSV) and investigate the protective immune response against RSV infection. Methods Recombinant plasmid DNA of pcDNA3.1~G was constructed by standard RT-PCR based cloning procedure. The immunogenicity of recombinant G protein transiently expressed in HEK293 cells was detected by Western blot. BABL/c mice were intramuscularly immunized with pcDNA3.1~G. Samples of lung, sera, bronchoalveolar lavage fluid(BALF) were collected before and after RSV challenge; virus titer in lung was detected by viral titration; sections of paraffin embedding lung tissues were stained by haematoxylin and eosin(HE) for histological analyses; sera anti-RSV IgG levels were examined by ELISA; Th1/Th2 cytokine were detected by ELISA kit, the T lymphocyte subsets of BALF was determined by immunefluorescence staining followed by flow cytometry. Results Plasmid DNA of pcDNA3.1~G was successfully constructed. The expressed target protein possesses immunogenicity. After challenge, pcDNA3.1~G immunized mice presented relieved pathological changes in lung as well as reduced lung viral titers. The RSV specific IgG was detected in sera of immunized mice. There was significantly increased number of CD25~+CD4~+ T cells in mice BALF. Conclusion We constructed a pcDNA3.1~G plasmid DNA vaccination which can induce evident protective cellular immunity against RSV infection in mice with the increased number of CD25~+CD4~+ T cell subpopulation.

plasmid can be used to study the pathogenic mechanism of target gene products of L.interrogans.

Objective To screen and identify the predominant T-and B-cell ( T-B) combined an-tigenic epitopes on the outer membrane protein Loa22 of pathogenic Leptospira interrogans ( L.interrogans) stains and to further analyze their immunogenicity.Methods PCR analysis was used to detect loa22 gene in L.interrogans strains belonging to eight different serogroups or serovars prevalent in China.The PCR prod-ucts were sequenced after T-A cloning.A prokaryotic expression system for loa22 gene of L.interrogans sero-group Icterohaemorrhagiae serovar Lai strain Lai was constructed.The expressed recombinant protein rLoa22 was extracted by Ni-NTA affinity chromatography.The rabbit anti-rLoa22 serum samples and IgG were pre-pared.The T-B combined antigenic epitopes on Loa22 protein were predicted by using professional bioinfor-matic softwares.Phage display in combination with Western blot assay and ELISA were performed to identify the immunogenicity of the recombinant phage PⅢ protein-displayed and artificially-synthesized T-B com-bined antigenic epitopes, respectively.MTS assay and ELISA were performed to detect the activation of T cells and the expression of IL-2, IL-4 and IFN-γinduced by T-B combined antigenic epitopes, respectively. Results All of the tested pathogenic Leptospira strains were positive for loa22 gene, sharing 85.5%-99.8%homologies in nucleotide sequences and 93.9%-99.5%homologies in amino acid sequences.The construc-ted prokaryotic expression system for loa22 gene efficiently expressed the rLoa22 protein.Among four T-B combined antigenic epitopes (Loa22-77, Loa22-90, Loa22-125 and Loa22-157), only Loa22-90 epitope presented a strong positive band in Western blot analysis.The proliferation of CD4+T cells and the expres-sion of IL-2 ( Th1 ) and IL-4 ( Th2 ) were significantly enhanced by the stimulation with Loa22-90 epitope peptide (P<0.05).Conclusion Loa22 protein is a sequence-conserved genus-specific outer membrane protein of L.interrogans.The Loa22-90 epitope is the predominant T-B combined antigenic epitope of Loa22 protein, which might be used as a candidate antigenic epitope in the development of multiple antigenic pep-tide ( MAP) vaccines against Leptospira infection.

Objective To investigate the application value of DWI based on biexponential signal decay model with extended b-fac-tor range in differential diagnosis of benign and malignant breast lesions.Methods A total of 57 patients with breast tumor under-went DWI based on the biexponential model with 12 b-factors (0,10,20,50,100,200,400,600,800,1000,1 200 and 1 500 s/mm2 ), including benign lesions in 1 9 patients (24 breast tumors,defined as benign group)and malignant ones in 38 (47 tumors,defined as malignant group ).The values of slow apparent diffusion coefficient,fast apparent diffusion coefficient and fraction of fast ADC of le-sions were measured at a workstation (Advantage Windows 4.5).Differences in these parameters between the benign and malignant groups were compared.Results The ADCslow,ADCfast and ffast were(1.434±0.291)×10 -3 mm2/s,(2.744±0.050)×10 -3 mm2/s and (0.677±0.130)% in benign group,and (0.614±0.196)×10 -3 mm2/s,(2.692±0.068)×10 -3 mm2/s and (0.446±0.112)% in malig-nant one,respectively.The statistical differences in ADCslow and ffast were found between two groups (P <0.05),whereas no difference in ADCfast was found.Conclusion Biexponential signal decay model of DWI with extended b-factor range can provide helpful tissue characterization parameters for the differential diagnosis of benign and malignant breast lesions.

Objective To summarize the nursing experience of allogeneic hematopoietic stem cell transplantation patients with circumferential mixed hemorrhoid intercurrent infection, explore the nursing methods of severe circumferential mixed hemorrhoid before allogeneic hematopoietic stem cell transplantation. Methods The detailed nurse plan and carefully observation and assess of lesion were performed daily. Iodophor water at the concentration of 0.5‰ was used for anal bath and 2% iodine ointment was used to smear perianal to prevent of infection. And gauzes soaked iodophor were used to cover the lesions, and Chinese traditional herb for anal bath and ultraviolet exposure to control infection. Additionally, the necessary psychological support was given to overcome the fear. Results After the careful nursing, the patient passed through the agranulocytosis period and achieved the success of transplantation. Conclusions This case suggests that the methods in prophylaxis of patients with circumferential mixed hemorrhoids is effectious.

Objective To isolate and identify the pathogenic bacteria from peripheral blood of a patient with septicemia of unknown etiology and to analyze their drug resistance genes.Methods Two peripheral blood samples were collected from the patient after having fever.Several assays including culturing bacteria on blood agar plates by using streaking technique,Gram-staining of bacterial colonies and microscopic observation,VITEK 2-compact automatic bacterium detection and analysis system as well as a sequencing analysis of the 16s rRNA gene were performed to identify the bacterial pathogens in blood samples.Microdilution test was performed to detect the drug susceptibilities of isolated bacteria to antibiotics.Confirmatory tests were performed to detect the production of β-lactamase and extended spectrum β-lactamase by the isolated strains and the phenotypes of AmpC enzyme and carbapenemase.PCR was used to identify the β-lactamase-encoding genes in the isolated strains by using the primers of 19 common β-lactamase-,AmpC enzyme-and carbapenemase-encoding genes in Enterobacteriaceae strains and the primers of 21 annotated gene sequences encoding the β-lactamase of a Ralstonia mannitolilytica strain.The PCR products were sequenced and analyzed after T-A cloning.Results Ralstonia mannitolilytica strains were isolated from the two peripheral blood samples.The isolated strains were sensitive to ceftriaxone,cefepime,ciprofloxacin,ofloxacin,tigecycline and compound sulfamethoxazole (SMZ-TMP),but resistant to the other 11 tested antibiotics.Results of PCR amplification by using the primers of common genes encoding β-lactamase of Enterobacteriaceae strains were all negative.Fragments of genes encoding the β-lactamase of the isolated Ralstonia mannitolilytica strain were successfully amplified,which were TK49_09850,TK49_12955,TK49_14470,TK49_14495and TK49_18990.The sequences of the amplified gene fragments were not similar to those of the common β-lactamase-encoding genes in Enterobacteriaceae strains.Conclusion The patient was infected with Ralstonia mannitolilytica.The isolated Ralstonia mannitolilytica strain showed a high-level drug resistance with a noticeable diversity against different β-lactam antibiotics.The genes encoding β-lactamase of the isolated Ralstonia mannitolilytica strain were completely different to those of Enterobacteriaceae strains.