Construction, expression and characterisation of a human anti-CD48 monoclonal antibody

Monoclonal antibodies and antibody-based entities are a major class of therapeuticproteins. The surge in development of monoclonal antibodies is predominantly due tothe utilization of technologies for producing fully human antibodies, namely phagedisplay and transgenic mice with human humoral immune systems. Human CD48, acell-surface adhesion molecule, is a potential tumor target for the treatment of whiteblood cell malignancies, principally leukemia and lymphoma. A truncated, secretedform of CD48 was expressed in CHO cells, and was shown to bind to existing anti-CD48 murine antibodies. A human anti-CD48 scFv antibody fragment, (designatedscFv-N2A) was previously isolated using phage display technology from a synthetichuman scFv immunoglobulin gene library. The scFv-N2A was reassembled as a humanIgG1 monoclonal antibody (designated IgG1-N2A), expressed in CHO cells and thebinding of IgG1-N2A to recombinant CD48 was confirmed by enzyme-linkedimmunosorbent assay, surface plasmon resonance and fluorescence activated cellsorting. IgG1-N2A binding to CD48 on Raji cells showed that the specificity of thehuman antibody for GPI-linked CD48 was conserved. In biological studies using ahuman lymphoma cell line (Raji), it was found that the IgG1-N2A antibody was able toinduce potent growth inhibition, with a 68% reduction in viable cells. Furthermore,Raji cells treated with IgG1-N2A showed evidence of increased ethidium bromideuptake and cell shrinkage, which are characteristics associated with direct induction ofapoptosis. The data suggests the novel human anti-CD48 IgG1-N2A monoclonalantibody can block proliferation and promote apoptosis of lymphoma cells, andtherefore has potential as a lead antibody candidate for the treatment of white blood cellmalignancies

Landslide Susceptibility Mapping Using Logistic Regression Analysis along the Jinsha River and Its Tributaries Close to Derong and Deqin County, Southwestern China

The objective of this study was to identify the areas that are most susceptible to landslide occurrence, and to find the key factors associated with landslides along Jinsha River and its tributaries close to Derong and Deqin County. Thirteen influencing factors, including (a) lithology, (b) slope angle, (c) slope aspect, (d) TWI, (e) curvature, (f) SPI, (g) STI, (h) topographic relief, (i) rainfall, (j) vegetation, (k) NDVI, (l) distance-to-river, (m) and distance-to-fault, were selected as the landslide conditioning factors in landslide susceptibility mapping. These factors were mainly obtained from the field survey, digital elevation model (DEM), and Landsat 4⁻5 imagery using ArcGIS software. A total of 40 landslides were identified in the study area from field survey and aerial photos’ interpretation. First, the frequency ratio (FR) method was used to clarify the relationship between the landslide occurrence and the influencing factors. Then, the principal component analysis (PCA) was used to eliminate multiple collinearities between the 13 influencing factors and to reduce the dimension of the influencing factors. Subsequently, the factors that were reselected using the PCA were introduced into the logistic regression analysis to produce the landslide susceptibility map. Finally, the receiver operating characteristic (ROC) curve was used to evaluate the accuracy of the logistic regression analysis model. The landslide susceptibility map was divided into the following five classes: very low, low, moderate, high, and very high. The results showed that the ratios of the areas of the five susceptibility classes were 23.14%, 22.49%, 18.00%, 19.08%, and 17.28%, respectively. And the prediction accuracy of the model was 83.4%. The results were also compared with the FR method (79.9%) and the AHP method (76.9%), which meant that the susceptibility model was reasonable. Finally, the key factors of the landslide occurrence were determined based on the above results. Consequently, this study could serve as an effective guide for further land use planning and for the implementation of development

Validation of Reference Genes for Gene Expression by Quantitative Real-Time RT-PCR in Stem Segments Spanning Primary to Secondary Growth in Populus tomentosa.

The vertical segments of Populus stems are an ideal experimental system for analyzing the gene expression patterns involved in primary and secondary growth during wood formation. Suitable internal control genes are indispensable to quantitative real time PCR (qRT-PCR) assays of gene expression. In this study, the expression stability of eight candidate reference genes was evaluated in a series of vertical stem segments of Populus tomentosa. Analysis through software packages geNorm, NormFinder and BestKeeper showed that genes ribosomal protein (RP) and tubulin beta (TUBB) were the most unstable across the developmental stages of P. tomentosa stems, and the combination of the three reference genes, eukaryotic translation initiation factor 5A (eIF5A), Actin (ACT6) and elongation factor 1-beta (EF1-beta) can provide accurate and reliable normalization of qRT-PCR analysis for target gene expression in stem segments undergoing primary and secondary growth in P. tomentosa. These results provide crucial information for transcriptional analysis in the P. tomentosa stem, which may help to improve the quality of gene expression data in these vertical stem segments, which constitute an excellent plant system for the study of wood formation

Laboratory evaluation of transgenic Populus davidiana×Populus bolleana expressing Cry1Ac + SCK, Cry1Ah3, and Cry9Aa3 genes against gypsy moth and fall webworm.

Transgenic poplar lines 'Shanxin' (Populus davidiana×Populus bolleana) were generated via Agrobacterium-mediated transformation. The transgenic lines carried the expression cassettes of Cry1Ac + SCK, Cry1Ah3, and Cry9Aa3, respectively. The expression levels of the exogenous insect resistance genes in the transgenic lines were determined by Q-PCR and Western blot. Leaves of the transgenic lines were used for insect feeding bioassays on first instar larvae of the gypsy moth (Lymantria dispar) and fall webworm (Hyphantria cunea). At 5 d of feeding, the mean mortalities of larvae feeding on Cry1Ac + SCK and Cry1Ah3 transgenic poplars leaves were 97% and 91%, while mortality on Cry9Aa3 transgenic lines was about 49%. All gypsy moth and fall webworm larvae were killed in 7-9 days after feeding on leaves from Cry1Ac + SCK or Cry1Ah3 transgenic poplars, while all the fall webworm larvae were killed in 11 days and about 80% of gypsy moth larvae were dead in 14 days after feeding on those from Cry9Aa3 transgenic lines. It was concluded that the transgenic lines of Cry1Ac + SCK and Cry1Ah3 were highly toxic to larvae of both insect species while lines with Cry9Aa3 had lower toxicity,and H. cunea larvae are more sensitive to the insecticidal proteins compared to L. dispar. Transgenic poplar lines toxic to L. dispar and H. cunea could be used to provide Lepidoptera pest resistance to selected strains of poplar trees

Effects of predictive nursing intervention on cognitive impairment and neurological function in ischemic stroke patients

Abstract Background Ischemic stroke is a clinical emergency caused by insufficient intracranial blood supply, which eventually leads to brain tissue necrosis and neurological impairment. Predictive nursing intervention has achieved impressive success in the nursing of multiple surgeries. However, the role of predictive nursing intervention in the care of patients with ischemic stroke remains unclear. Methods This study was a randomized controlled trial. Based on the inclusion and exclusion criteria, 126 patients were randomly assigned into two groups, namely the control group and the predictive nursing intervention group. Both groups were treated with thrombolytic therapy with alteplase. The patients in the control group were given routine nursing intervention and the predictive nursing intervention group received additional predictive care. Neurologic functions and cognitive impairment were evaluated by National Institutes of Health Stroke Scale (NIHSS), Fugl‐Meyer assessment (FMA), Montreal cognitive assessment (MoCA), and mini‐mental state examination (MMSE) scales, respectively. Door‐to‐Needle Times, venous thromboembolism (VTE)‐related parameters, and complications were recorded. Results Predictive nursing intervention significantly shortened the Door‐to‐Needle Times and enhanced the peak/average femoral venous blood flow and femoral venous diameter. In addition, predictive nursing intervention improved the NIHSS, FMA, MMSE, and MoCA scores and remarkably reduced the recurrence of ischemic stroke, deep vein thrombosis and gingival bleeding. Conclusion Predictive nursing intervention is beneficial to improve the effects of thrombolytic therapy in patients with ischemic stroke, which improves the neurological, cognitive and motor functions of patients, and reduces the occurrence of complications, suggesting an important clinical application value