Objective The study was to investigate the feasibility of using an intravascular Loopless Monopole Antenna (ILMA) for 3.0 T MR imaging of water bath and deep-seated arterial walls of experimental animal.Methods A novel intravascular loopless monopole antenna (ILMA) was developed,including a non-shield loach guide-wire and a matching circuit.The non-shield loach guide-wire is used as a receive antenna,with the diameter of 0.019 in( 1 in =2.54 cm) and length of 23.11 in.During the MR scanning,the ILMA was used as a receive-only probe,while body coil was used to transmit the RF pulses.Utilizing the coil in water bath and in-vivo animal experiment,we measured signal-to-noise ratio (SNR) and contrast-to-noise ratio(CNR) of artery wall using the same scanning parameter compared with phased-array coil.Results In the study,the developed novel ILMA conduced to improved SNR of imaging and much higher space resolution( 313 μm).First,the feasibility of acquiring the wall images was demonstrated on phantoms.The SNRs map generated by the matlab software showed that in comparison with the phased-array coil,ILMA generated higher SNR of the phantom wall when using the same sequences,parameters,and slices (86.8 ±0.8 vs.9.9 ±0.1,P ＜0.01 ).When imaging the aorta wall with the ILMA and phased-array coil,the SNRs of the arterial wall with the ILMA is 60.4 ±20.9,61.3 ±22.5,59.8 ±20.4,32.3 ±22.6 (T1WI),51.2 ±21.6,49.8 ± 15.5,50.4 ± 17.2,22.4 ± 18.3 (T2WI),the CNRs of the aorta wall with theILMA is 19.8±8.1,18.9±9.2,19.6±11.8,20.7 ± 13.3(T1WI),17.7±6.4,18.6±6.9,17.2 ± 6.4,17.2 ± 6.4 ( T2 WI),compared with phased-array coil,t values SNR:6.36,3.84,3.51,6.92(T1 WI),3.47,4.89,6.35,4.21 (T2WI),CNR:3.56,3.97,-0.71,4.74 (T1WI),3.99,3.01,4.27,5.03(T2 WI,P ＜ 0.05 ),respectively.Conclusion The study demonstrates the capability of using an MR ILMA to generate 3.0 T MR in-vivo experiments,the developed novel ILMA conduces to increased SNR compared with the conventional phased-array coil.

Objective Explore the change of IL-1β and IL-18 expression in pulmonary hypertension induced by monocrotaline.Methods Divide the mouses into two groups, control group and experimental group (n=10).Establish rats pulmonary hypertension model induced by monocrotaline.Detect the model by ultrasound, myocardial cells HE dyeing and tunnel test;ELISA was used to detect the serum biological markers NF-κB, COX2, IL-6, IL-1β, TNF-α and NO;Immunohistochemical was used to detect the expression level of IL-1β and IL-18 in the lung tissue;the protein change of NLRP3 in the lung tissue was detected by Western blot.Results Serum biological markers of NF-κB, COX2, IL-6, IL-1β, TNF-α and NO are significantly increased in PAH rats(P<0.05);The expression of IL-1β, IL-18 in the lung tissue increased obviously(P<0.05);The NLRP3 protein expression was significantly higher in experimental group.Conclusions Changes of NLRP3 effect increase expression of IL-1β and IL-18and which may play an important role in pulmonary hypertension induced by monocrotaline.

Pulldown assay is an in vitro method for studies of protein-protein interactions, in which tagged proteins are usually expressed as the bait to enrich other proteins that could bind to them. In this technology, the GST tag is broadest used for its modest size and hydrophilic property. In most cases, the GST tag could increase the hydrophility of the fusion protein and help to avoid the formation of inclusion bodies. However, in the other few cases, the target protein may be strongly hydrophobic or have complicated structures that were hard to fold and assemble in correct conformations without champerons, and even the existence of GST tag could not make them soluble. These proteins were always expressed as inclusion bodies and had no functions. LMO2 was a small molecular weight and insoluble protein, in this study, GST system and MBP system were used to express GST-LMO2 and MBP-LMO2 fusion proteins, respectively. We found that GST-LMO2 fusion protein was expressed as inclusion bodies whereas MBP-LMO2 fusion protein was expressed in soluble form. Moreover, the production rate of MBP-LMO2 was also much higher than GST-LMO2. Then MBP-LMO2 fusion proteins and renatured GST-LMO2 fusion proteins were used as bait in pulldown assay to study the interaction between LMO2 and endogenous GATA1 in K562 cells. Western blot analyses showed that both of these proteins could bind to endogenous GATA1 in K562 cells, but recovered GATA1 protein by MBP-LMO2 fusion protein was much more than GST-LMO2 fusion protein. These results suggest that using of MBP system is a helpful attempt in the case of studying small molecular weight, strong hydrophobic proteins.
Adaptor Proteins, Signal Transducing ; Carrier Proteins ; chemistry ; Chemical Precipitation ; DNA-Binding Proteins ; chemistry ; GATA1 Transcription Factor ; chemistry ; Genetic Vectors ; Glutathione Transferase ; chemistry ; Humans ; K562 Cells ; LIM Domain Proteins ; Maltose-Binding Proteins ; Metalloproteins ; chemistry ; Protein Binding ; Protein Interaction Domains and Motifs ; Protein Renaturation ; Proto-Oncogene Proteins ; chemistry ; Recombinant Fusion Proteins ; genetics ; metabolism

Bone morphogenetic protein 2 (BMP2), which belongs to the transforming growth factor-beta (TGF-beta) superfamily, is a multifunctional molecule with distinct abilities to induce bone formation. BMP2 has been identified to have eminent pharmaceutical importance for clinical application. We previously constructed stable cell line in Chinese hamster ovary cells (CHO) that highly expressed recombinant human BMP2 (rhBMP2). For large-scale production of the recombinant protein used in clinical application, it is critical to have both high expression and stability of the protein. In the present study, the stability of the cell line (rCHO(hBMP2)-C8) with the highest expression, as well as the stability of rhBMP2 protein were investigated systematically. We cultured the rCHO (hBMP2)-C8 cell line in the presence or absence of MTX for two months, the cell growth and rhBMP2 production characteristics were examined during the culture; we found the duration that the rCHO(hBMP2)-C8 cell line could secret rhBMP2 continually into the serum-free medium. Moreover, we detected the temperature sensitivity of rhBMP2 in culture medium. This study will contribute to our understanding for further producing rhBMP2 by large-scale culture technology.
Animals ; Bone Morphogenetic Protein 2 ; biosynthesis ; genetics ; CHO Cells ; Cell Culture Techniques ; methods ; Cricetinae ; Cricetulus ; Culture Media ; Genetic Vectors ; genetics ; Humans ; Recombinant Proteins ; biosynthesis ; genetics

Objective: To elaborate the characteristics and advantages of Whole-Mount immune fluorescence staining by observing the lymphatic vessels of mice. Methods: The ear skin tissue, the hindlimb lymphatic vessels and the mesenteric lymphatic vessels were harvested from normal C57 mice. The tissue samples were subjected to whole-tissue immunofluorescence staining.These tissue samples were fixed by paraformaldehyde, blocked by bovine serum and incubated in primary and secondary antibodies. Then, the lymphatic vessels were observed and analyzed in these samples with a confocal laser-scanning microscope. Results: The capillary lymphatic vessels and lymphatic endothelial cells can be clearly showed in the ear skin. The valves and smooth muscles can be clearly showed in the hindlimb and mesenteric lymphatic vessels by Whole-Mount immunofluorescence staining. Conclusions The whole-tissue immunofluorescence staining technique can observe the external morphology of lymphatic vessels clearly and stereoscopically, and can deeply observe the internal structure of lymphatic vessels. This technique can provide more accurate study on physiology and pathology of lymphatic vessels.

In May 2019, the International Diabetic Foot Working Group (IWGDF) updated and issued guidelines for the prevention and management of diabetic foot disease. This guide puts forward some suggestions for the diagnosis, treatment and effective prevention of diabetic foot: the prevention of diabetic foot should start with high-risk foot, early screening and treatment of diabetic foot infection, foot ulcer and peripheral vascular disease and early comprehensive treatment. Effective prevention and early treatment can reduce the incidence of cardiovascular and cerebrovascular events in patients with diabetes, reduce the amputation rate and mortality, and improve the prognosis and quality of life of patients.