Dengue virus (DENV) envelope [E] protein is the major surface protein of the virions that indued neutralizing antibodies. The domain III of envelope protein (EDIII) is an immunogenic region that holds potential for the development of vaccines; however, the epitopes of DENV EDIII, especially neutralizing B-cell linear epitopes, have not been comprehensively mapped. We mapped neutralizing B-cell linear epitopes on DENV-1 EDIII using 27 monoclonal antibodies against DENV-1 EDIII proteins from mice immunized with the DENV-1 EDIII. Epitope recognition analysis was performed using two set of sequential overlapping peptides (16m and 12m) that spanned the entire EDIII protein from DENV-1, respectively. This strategy identified a DENV-1 type- specific and a group-specific neutralizing epitope, which were highly conserved among isolates of DENV-1 and the four DENV serotypes and located at two regions from DENV-1 E, namely amino acid residues 309-320 and 381-392(aa 309-320 and 381-392), respectively. aa310 -319(310KEVAETQHGT319)was similar among the four DENV serotypes and contact residues on aa 309 -320 from E protein were defined and found that substitution of residues E309 , V312, A313 and V320 in DENV-2, -3, -4 isolates were antigenically silent. We also identified a DENV-1 type-specific strain-restricted neutralizing epitope, which was located at the region from DENV-1 E, namely amino acid residues 329-348 . These novel type- and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.
Amino Acid Sequence ; Animals ; Antibodies, Neutralizing ; immunology ; Dengue ; virology ; Dengue Virus ; chemistry ; genetics ; immunology ; Epitope Mapping ; Epitopes, B-Lymphocyte ; chemistry ; genetics ; immunology ; Humans ; Mice ; Molecular Sequence Data ; Viral Envelope Proteins ; chemistry ; genetics ; immunology

OBJECTIVETo study the therapeutic effects of posterolateral depression fractures of the tibial plateau through a modified anterolateral approach.

METHODSFrom February 2011 to January 2012,13 patients with posterolateral depression fractures of the tibial plateau were treated through a modified anterolateral approach. There were 8 males and 5 females, ranging in age from 28 to 59 years old (49.2 years old on average). Data from patients were collected retrospectively as follows: X-ray, time of fracture healing and the complications of fracture healing. The patients were evaluated both clinically and radiologically according to the Rasmussen score system.

RESULTSAll the patients were followed up, and the duration ranged from 6 to 18 months (mean 13.7 months). All the patients got bony union. The average radiographic bony union time was 15.1 weeks (ranged, 11 to 17 weeks). No case of secondary articular depression was found. No complications such as malunion or joint stiffness were found. But 1 patient had superficial infection and 1 patient had common peroneal nerve injury. According to the Rasmussen score system,the mean radiological score was 16.50 ± 0.67 (ranged, 13 to 18), and the mean functional score was 25.20 ± 2.21 (ranged, 13 to 30). The mean range of knee motion was (125.3 ± 9.3)° (ranged, 0° to 135°).

CONCLUSIONTreatment of depression fractures of posterolateral tibial plateau with a modified anterolateral approach is a safe method with effective exposure, due to its stable fixation and relatively good outcome with minimal soft-tissue complications. It is regarded as an ideal procedure for depression fractures of posterolateral tibial plateau.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery

Aged ; Cardiac Pacing, Artificial ; Cardiac Resynchronization Therapy ; Female ; Heart Ventricles ; Humans

Effect of p-nitrophenol shock on microbial populations and sludge activity in UASB reactor was studied by DGGE-PCR of 16S rDNA fragments and detection of COD removing and biogas yield.The results showed that p-nitrophenol seriously inhibited the sludge activity,resulting in the drop of biogas and COD removing rate.The 40mg/L p-nitrophenol had more inhibition than 20mg/L p-nitrophenol.It would take 27 and 16 days respectively for reactor to recover after 40mg/L and 20mg/L p-nitrophenol shock.The diversity of eubacteria and methanogens were also effected by the p-nitrophenol shock.The variation of eubacteria was more than that of methanogens after p-nitrophenol shock.The drop of biogas was mainly related to the variation of Methanosaeta sp.and Methanomicrobia sp.after p-nitrophenol shock.Among the eubacteria the population of Chloroflexi sp.、Bacteroide sp.and Anaerovibrio sp.decreased greatly after p-nitrophenol shock.And more,the Rheinheimera sp disappeared after 40mg/L p-NP treatment.But the Flavobacteria sp.appeared after p-nitrophenol shock,which was probably related to the degradation of p-NP.

OBJECTIVEGas chromatography (GC) was used to investigate the cellular fatty acid (CFA) composition of 141 Acinetobacter baumannii and 32 A. calcoaceticus isolates from different locations in China and to find chemical markers to differentiate these two closely related bacteria.

METHODSWhole cell fatty acid methyl esters (FAMEs) were obtained by saponification, methylation, and extraction for GC analysis, followed by a standardized Microbial Identification System (MIS) analysis.

RESULTSAll A. baumannii and A. calcoaceticus strains contained some major fatty acids, namely, 18:1 ω9c, 16:0, Sum In Feature 3, 12:0, 17:1ω8c, 3-OH-12:0, 17:0, Sum In Feature 2, 2-OH-12:0, and 18:0 compounds. Although most of the total CFAs are similar between A. baumannii and A. calcoaceticus strains, the ratios of two pairs of CFAs, i.e., Sum In Feature 3/18:1 ω9c versus 16:0/18:1 ω9c and Sum In Feature 3/18:1 ω9c versus unknown 12.484/18:1 ω9c fatty acids, could differentiate these two closely related bacteria. A. baumannii could be easily classified into two subgroups by plotting some ratios such as Sum In Feature 3/16:0 versus 17:0 and Sum In Feature 3/2-OH-12:0 versus 17:0 fatty acids.

CONCLUSIONThe ratios of some CFAs could be used as chemical markers to distinguish A. baumannii from A. calcoaceticus.

Acinetobacter baumannii ; classification ; cytology ; metabolism ; Acinetobacter calcoaceticus ; classification ; cytology ; metabolism ; Biomarkers ; metabolism ; Fatty Acids ; metabolism ; Species Specificity

ObjectiveAnalyzing the left atrium and pulmonary vein morphologicallv by 64 multislice spiral CT(MSCT)scan to guide the catheter ablation of Atrial fibrillation.MethodsTwo hundred and thirty-two patients(146 cases in atrial fibrillation group and 86 cases in control group)received 64 MSCT examination of the left atrium and pulmonary vein.The incidence of anatomical variation of pulmonary vein was compared between atrial fibrillation group and control group. For each group,the anatomical morphology ot every pulmonary vein and the auricle of left atrium was analyzed, the diameter of the orifice of each pulmonary vein and the size of left atrium were measured.ResultsSixty-four MSCT of left atrium and pulmonary vein could demonstrate detailed connecting type between left atrium and pulmonary Veins and the possible anatomieal variation. Anatomical variation of pulmonary vein in this study accounted for 16.8% (39/232)of total sample. For both groups,orifices of pulmonary veins appeared oval and their superoinferior diameters were larger than their anteroposterior diameters. There was significant difference in the inner diameter of left atrium between atrial fibrillation group and control group[atrial fibrillation group:(39.47±8.98)mm,control group:(36.94 ±5.49)mm,P=0.02],while there was no difference in the diameters of orifices ot puhnonary veins between two groups [ superoinferior diameters of pulmonary veins in atrial fibrillation group:left-up(18.15±1.35)mm,left-down(16.96 ±1.18)mm,right-up(17.50±

Background Cooking oil fumes are closely related to immune response, and adipose tissue also plays an important role in immune regulation. At present, the biological effect and mechanism of inflammation of adipose tissue induced by oil fume exposure are not clear yet. Objective To investigate the inflammatory effect of different exposure duration of cooking fumes on adipose tissue in mice and explore the role of Nod-like receptor pyrin domain 3 (NLRP3)/cysteinyl aspartate specific proteinase 1 (Caspase 1)/interleukin (IL)-1β signaling pathway. Methods Forty 8-week-old female C57BL/6J mice were randomly divided into 3-day control group (CON₃ group), 7-day control group (CON₇ group), 3-day oil fume exposure group (COF₃ group), and 7-day oil fume exposure group (COF₇ group), with 10 mice in each group. The mice were exposed to oil fumes in a cooking oil fume formation and exposure equipment (COFFEE) for 20 min, followed by a 10-min pause, 1 h a day for consecutive 3 d or 7 d. General condition of mice was observed and body weight was measured every day. After exposure, blood was sampled from the eyeball. Serum levels of IL-6, IL-27, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA). The adipose tissue of mice was collected and observed after hematoxylin-eosin (HE) staining. The percentages of CD4⁺ and CD8⁺T cells in adipose tissue were detected by flow cytometry. Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of nuclear factor-κB (NF-κB), NLRP3, Caspase 1, and IL-1β in adipose tissue. Western blot was used to detect the expression levels of NLRP3, Caspase 1, and IL-1β in adipose. Results Compared with the corresponding control group, serum IL-6, IL-27, and IL-1β contents in the COF₃ group and the COF₇ group were significantly increased (P<0.05) except IL-6 in the COF₃ group, and the levels in the COF₇ group were significantly higher than those in the COF₃ group (P<0.05). Vacuolar lipid droplets in adipocytes decreased, cytoplasm shrank, and inflammatory cells infiltrated in the COF₇ group after HE staining. The flow cytometry results showed that the proportions of CD4⁺ and CD8⁺T cells in adipocytes of the COF₃ group and the COF₇ group were increased compared to the corresponding control group, with a significant increase in the COF₇ group (P<0.05), and the CD4⁺/CD8⁺T ratio also significantly increased progressively in the two groups (P<0.05). The results of RT-qPCR showed that compared with the corresponding control group, the mRNA expression levels of NF-κB, NLRP3, Caspase 1, and IL-1β in adipose tissue of mice in the COF₃ group and the COF₇ group were significantly increased (P<0.05, P<0.01). The mRNA expression levels of mice in each exposure group gradually increased over time. The Western blot results showed that compared with the corresponding control group, the protein expressions of NLRP3 and Caspase 1 in the COF₃ group were significantly increased (P<0.01), and the expression of IL-1β protein also increased but without statistical significance. The protein expressions of NLRP3, Caspase 1, and IL-1β in the COF₇ group were significantly higher than those in the CON₇ group (P<0.05, P<0.01). Conclusion Acute exposure to cooking oil fumes can induce significant inflammatory response in adipose tissue, and the effect gradually increases with the extension of exposure time. The mechanism of action may be related to the activation of NLRP3 inflammasome signaling pathway.

Objective To evaluate serum soluble intercellular adhesion molecule-1 (sICAM-1) before and after lung transplantation for diagnosing acute rejection. Methods Biotin-streptavidin time resolved fluoroimmunoassay (BSA-TRFIA) was used to detect the concentration of serum sICAM-1 before and after lung transplantation in 26 patients. All patients were divided into stable lung transplantation group (n =16), acute rejection group (n =4) and infected group (n =6). The serum level of sICAM-1 in those groups was compared with that of the control group ( n = 30 ) by the non-parametric rank sum test ( KruskalWallis H test). Results No significant difference was found for serum sICAM-1 among the three groups and the control group before operation: (357.07 ± 220.01 ), ( 396. 18 ± 136.25 ), (468.95 ± 85.48 ) μg/L vs(348.63 ±69. 12) μg/L, H=6. 0436, P ＞0.05. However, when rejection and infection happened after operation, the serum sICAM-1 increased in the acute rejection group (455.53 ± 126.51 μg/L) and decreased in the infection group (146.43 ± 327.11 μg/L), and the level in the stable transplantation group was (274.23 ± 157.53 ) μg/L (H = 21. 8994, P ＜ 0.01 ). Conclusion Serum sICAM-1 level might be a potential marker to differentiate acute rejection from infection after lung transplantation.

CD4-Positive T-Lymphocytes ; immunology ; CD56 Antigen ; analysis ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; immunology ; Dendritic Cells ; immunology ; Humans ; Liver Neoplasms ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo study the gap junction mechanisms for synergistic effects of liuwei di-huang pill (LDP) containing serum on HSV-tk/GCV suicide gene therapy of mouse malignant melanoma B16 cells.

METHODSThe LDP containing serum (2.5% LDP serum group, 5.0% LDP serum group, and 10.0% LDP serum group) and the control serum group were set up. The effects of LDP on mRNA expressions of Cx26 and Cx43 in mouse malignant melanoma B16 cells were detected by RT-PCR assay. The effects of LDP on protein expressions of Cx26 and Cx43 in B16 cells were detected using Western blot and indirect immunofluorescence assay. The interference efficiencies of Cx26-309, Cx26-337, Cx26-367, Cx26-2098 SiRNA on Cx26 gene in B16 cells were detected using RT-RCR technique. Cx26-2098 SiRNA, due to the optimal interference efficiency, was chosen to interfere Cx26 gene, and the bystander effect of LDP + HSV-tk/GCV was observed. The effects of the gap junctional intercellular communication (GJIC) inhibitor glycyrrhetinic acid on the killing of LDP + HSV-tk/GCV system to B16 cells were detected by MTT assay. In this experiment, the control serum group, 2.5% LDP serum group, 5. 0% LDP serum group, 10.0% LDP serum group, the control serum combined GCV group, 2.5% LDP serum combined GCV group, 5.0% LDP serum combined GCV group, 10.0% LDP serum combined GCV group, 2.5% LDP serum combined GCV + glycyrrhetinic acid group, 5.0% LDP serum combined GCV + glycyrrhetinic acid group, 10.0% LDP serum combined GCV + glycyrrhetinic acid group were set up. The final concentration of GCV was 20 micromol/L. The final concentration of glycyrrhetinic acid was 50 micromol/L.

RESULTSLDP containing serum could increase the protein and mRNA expressions of Cx43 in B16 cell in a dose-dependent manner. It had bi-directional regulation on the Cx26 protein and mRNA expressions. The low dose LDP had inhibition while high dose LDP could up-regulate its expression. After SiRNA interfered Cx26 gene, there was no obvious change in the bystander effect of LDP combined suicide gene therapy between before and after interference. There was significant reduction in the inhibition rate between before (48.75%, 59.39%, and 69.28%) and after blockage (29.14%, 38.71%, and 58.13%) of glycyrrhetinic acid in the 2. 5%, 5.0%, 10.0% LDP serum combined GCV groups (P <0.01).

CONCLUSIONThe synergistic effects of LDP containing serum on HSV-tk/GCV suicide gene therapy in mouse malignant melanoma B16 cells were correlated with the gap junction mechanisms, which might be achieved through increasing the expressions of Cx26 and Cx43.

Animals ; Cell Line, Tumor ; Connexin 26 ; Connexin 43 ; metabolism ; Connexins ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Gap Junctions ; metabolism ; Genetic Therapy ; Male ; Melanoma ; therapy ; Mice ; Rats, Sprague-Dawley ; Serum