Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.

Objective: To evaluate the clinical value of multislice CT (MSCT) in the diagnosis and preoperative TNM staging of gastric carcinoma. Methods: Fifty patients with advanced gastric carcinoma were examined by MSCT, gastrointestinal series (GI), fiberoptic gastroscopy (FG) and transabdominal ultrasonography (US). The results of the 4 methods were compared with postoperative pathological results. Forty patients, who were diagnosed as having advanced gastric carcinoma by both MSCT and US, had their TNM staging evaluated and the results were compared with postoperative pathological TNM evaluation. Results: The detection rates of MSCT, FG, GI and US for advanced gastric carcinoma were 98%, 100%, 88% and 80%, respectively. The detection rate of MSCT was not significantly different with that of FG, but was obviously higher than that of GI (P=0.027) and US (P=0.004). The accuracy of MSCT in preoperative TNM staging was significantly higher than that of US(92.5% vs 72.5%). Conclusion: MSCT, with two-phase thin slice incremental scanning image, multiplaner reformats (MPR) and three-dimension (3D) image, is more advantaged in detecting the gross type, size, location, invasion and metastasis of advanced gastric carcinoma, thus greatly improving the detection rate and preoperative TNM staging of advanced gastric carcinoma.

In order to develop an anti-FMDV A Type monoclonal antibo by (mAb),BABL/c mice were immunized with FMDV A type.Monoclonal antibodies (mAbs) 7B11 and 8H4 against Foot-and-mouth disease virus (FMDV) serotype A were produced by fusing SP2/O myeloma cells with splenocyte from the mouse immunized with A/AV88.The microneutralization titer of the mAbs 7B11 and 8H4 were 1024 and 512,respectively.Both mAbs contain kappa light chains,the mAbs were IgG1.In order to define the mAbs binding epitopes,the reactivity of these mAbs against A Type FMDV,were examined using indirect ELISA,the result showed that both mAbs reacted with A Type FMDV.These mAbs may be used for further vaccine studies,diagnostic methods,prophylaxis,etiological and immunological research on FMDV.Characterization of these ncindicated that prepared anti-FMDV A mAbs had no cross-reactivity with Swine Vesicular Disease (SVD) or FMDV O,Asial and C Type antigens.Their titers in abdomen liquor were 1:5×106 and 1:2×106,respectively.7B11 was found to be of subtype IgG1,8H4 was classified as IgG2b subtype.The mAbs prepared in this study,are specific for detection of FMDV serotype A,and is potentially useful for pen-side diagnosis.

Objectives To investigate the relationship between atherosclerotic acute myocardial infarction (AMI),acute cerebral infarction (ACI)and homocysteine (Hcy). Methods Three hundred and twenty consecutive patients with primary acute myocardial infarction (AMI)(group A)were admitted to the Department of Cardiology,310 patients with primary large artery atherosclerotic cerebral infarction (group B)were admitted to the Department of Neurology,and 327 healthy individuals without cardiovascular and cerebrovascular diseases (group C)at the Department of Physical Examination,Xuanwu Hospital, Capital Medical University were enrolled retrospectively from March 2010 to October 2011. The age and sex were matched in the 3 groups. All the clinical data of subjects were colleted in detail and then were compared and analyzed. Results (1)The Hcy levels (μmol/L)of group A,B,and C were 15. 10 (12. 43, 19.47),15. 80 (13. 10,20. 83),and 13. 20 (11. 00,16. 50;median [interquartile range]),respectively. There were significant differences among the 3 groups (P<0. 05). The incidences of hyperhomocysteinemia (HHcy)were 92. 8%(n=297),97. 1%(n=301),and 84. 7%(n=277)(P<0. 05). (2)Multivariate Logistic regression analysis showed that the independent risk factors for ACI were HHcy (OR 8. 97,95% CI 3. 01-26. 71),hypertension,diabetes,hyperlipidemia and blood ureanitrogen;the independent risk factors for AMI were HHcy (OR 4. 36,95% CI 1. 70-11. 21),hypertension,diabetes,hyperlipidemia,and total blood cholesterol. Conclusion HHcy is an independent factor for ACI and AMI,which have closer relationship with ACI. ACI and AMI have some common risk factors,but their degrees of action are different.

Adult ; Female ; Humans ; Lead ; toxicity ; MMPI ; Male ; Middle Aged ; Occupational Exposure ; Personality ; drug effects

Objective To analyze the clinical effects of the diverse prevention strategies on donation after citizen's death (DCD) donor fungal infection.Methods A retrospective study was performed on the clinical data of the antifungal preventive scheme to 261 DCD donors from January 2015 to August 2015 (the first period) and September 2015 to December 2016 (the second period) in Tongji Hospital.During the first period,the donors were administrated by ICU doctors and the antifungal agents were not applied as routine.The processes of organ procurement and trim were in accordance with the past experience.During the second period,the donor maintenance was reinforced,including antifungal preventive scheme,aseptic manipulation of organ procurement and rinsing and immersing allografts with broad-spectrum antibiotics or diluting povidone-iodine solutions during organ trim.Microbial culture specimens were performed in 1 574 samples including blood samples,perfusion fluids and arterial tissues and the pathogen distribution at the different periods was identified.Result In the microbial culture specimens of 1 574 samples,907 strains of pathogens were detected,including 799 strains of bacteria and 108 strains of fungi.The positive rate of fungi was 12.0％ (108/907) of all pathogens,and 17.3％ (108/626) of fungal cultures specimens.The fungi positive rate in the second period (13.6％,59/433) was significantly lower than that in the first period (25.4％,49/193,P＜ 0.05).Conclusion It is essential to reduce the incidence of donor fungal infection by increasing the microbial cultures and antifungal preventive scheme.And it is necessary that the organ procurement organization team enhances the risk awareness of donor-derived fungal infection and improves the aseptic manipulation of organ procurement.Additionally,rinsing and immersing allografts with broad-spectrum antibiotics or diluting povidone-iodine solutions may be a better option for preventing the donor-derived fungal infection during the allograft trim.

BACKGROUNDEffects of icariin on airway inflammation in asthmatic rats and the intervention of LPS induced inflammation are interfered with the machanism of icariin. Our study aimed to observe the effect of icariin on ovalbumin-induced imbalance of Th1/Th2 cytokine expression and its mechanism.

METHODSSixty male SD rats were randomly divided into control group (PBS), asthma group (ovalbumin (OVA)-induced), dexamethasone group, and OVA+icariin low, medium and high dose groups (5, 10, 20 mg/kg, respectively). Each group had ten rats. The model of OVA sensitization was a rat asthma model. Enzyme-linked immunosorbent assay (ELISA) method was used to observe the effects of icariin on interleukin-4 (IL-4) and inerferon γ (IFN-γ) in rats' lung tissue. Immunohistochemical staining was applied to detect the intervention effects of icariin on T cells (T-bet) and gatabinding protein 3 (GATA-3) in rat pulmonary tissue. Realtime RT-PCR was used to observe the intervention effects of icariin on T-bet and GATA-3 mRNA expression in rat pulmonary tissue and spleen lymphocytes. Western blotting was used to observe the icariin intervention effects on T-bet, GATA-3 and nuclear factor-Kappa B (NF-κB) p65 protein expressions in rat pulmonary tissue.

RESULTSThe ELISA results from pulmonary tissue showed that IL-4 expression was significantly reduced (P < 0.05), while the IFN-γ expression increased but not significantly when we compared OVA+icariin medium and high dose groups with the asthma group. Immunohistochemical staining of pulmonary tissue showed that the GATA-3 decreased significantly while the T-bet staining did not change in the OVA+icariin high dose group. In pulmonary tissue and spleen lymphocytes T-bet and GATA-3 mRNA expressions were significantly reduced (P < 0.05) in icariin treatment groups compared with the asthma model group. GATA-3 and T-bet mRNA in rat spleen lymphocytes in the asthma group were higher than in the control group. GATA-3 mRNA expression in pulmonary tissue significantly decreased (P < 0.05) while T-bet mRNA expression decreased but not significantly in the icariin treatment group compared with the asthma group. T-bet and GATA-3 protein expressions in pulmonary tissue increased significantly compared with the asthma group, which meant that icariin could inhibit the increase of GATA-3 protein, but not of T-bet. The bronchus, blood vessels and periphery pulmonary tissue had infiltration of inflammatory cells in the OVA+icariin high dose group while NF-κB p65 cells were reduced, and expression of NF-κB p65 in this group was less than in the asthma group. The expression of total p65 protein decreased with icariin treatment while the expression of cytoplasmic p65 protein increased.

CONCLUSIONSIcariin could regulate the imbalance of Th1/Th2 cytokines in asthmatic rat pulmonary tissue. Icariin could regulate the imbalance of Th1/Th2 associated transcription factors T-bet and GATA-3 in asthmatic rat pulmonary tissue and spleen lymphocytes. Icariin could inhibit the activation of NF-κB p65 protein in asthmatic rat pulmonary tissue.

Animals ; Asthma ; drug therapy ; immunology ; metabolism ; Blotting, Western ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Flavonoids ; therapeutic use ; GATA3 Transcription Factor ; metabolism ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lung ; metabolism ; Male ; Ovalbumin ; metabolism ; Polymerase Chain Reaction ; Rats ; Rats, Sprague-Dawley ; T-Box Domain Proteins ; metabolism ; Th1 Cells ; drug effects ; metabolism ; Th2 Cells ; drug effects ; metabolism ; Transcription Factor RelA ; metabolism

Purpose: Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear. Materials and Methods: The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2. Results: LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects. Conclusion This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.

Purpose: Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear. Materials and Methods: The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2. Results: LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects. Conclusion This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.

Purpose: Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear. Materials and Methods: The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2. Results: LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects. Conclusion This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.