Vaccines and antibodies are currently effective intervention strategies for preventing and treating COVID-19. Angiotensin-converting enzyme 2 (ACE2), a primary binding receptor for SARS-CoV-2, is a potential target for the prevention and treatment of COVID-19. At present, therapeutics based on the strategies that block the binding of SARS-CoV-2 Spike protein to the ACE2 receptor have entered clinical trials. On December 5, 2022, the journal Nature published the results of a pharmacological intervention to downregulate ACE2 expression for prevention of SARS-CoV-2 infections. The study found that a nuclear receptor, farnesoid X receptor for bile acids, regulated ACE2 expression. The clinical drug ursodeoxycholic acid that was used to treat liver diseases could downregulate ACE2 expression and prevent SARS-CoV-2 infections, showing a new potential pathway in managing COVID-19.
Objective To evaluate the osteogenesis of bi phasic ceramic-l ike biologic bone (BCBB) with tissue engineering in repairing segmental bone defects. Methods BMSCs isolated from the femoral and tibial marrow of 2-weekold Japanese white rabbit were cultured to passage 3. Then 20 μL of the cell suspension at a concentration of 1 × 107 cells/mLwere seeded into 15 mm × 15 mm × 5 mm BCBB block; the construction of tissue engineered BCBB was completed after 8 days of compound culture. Forty-eight adult Japanese white rabbits were randomly divided into groups A, B, C and D, then BCBBs cultured with BMSCs in vitro for 8 days (group A) and only BCBBs without BMSCs (group B) were respectively implanted into the radius segmental bone defects of rabbits, autogenous il iac bone graft (group C) and empty defect (group D) were used as controls. The specimens were examined after 4, 8, 12 and 24 weeks, the osteogenesis was evaluated through X-ray radiograph and histology examination. Results X-ray examination: the border between the material and host’s bone was clear after 4 weeks, and blurred after 8 weeks in group A and group B; the density of some part of the edge of the material was similar to that of radius and there was high density imaging in the materials of group A after 12 weeks; there was much high density imaging in the materials of group B after 12 weeks. The medullary cavity of bone was formed and l ittle high density imaging in the materials of group A after 24 weeks. Some high density imaging still existed in the materials of group B after 24 weeks. The X-ray evaluated scores showed that the scores of group A was higher than that of group B, and there was significant difference between group A and group B after 12 and 24 weeks (P lt; 0.05). Histological examination: there was new bone formation in the materials and also new bone grew adhesively on the surface of BCBB in group A. While in group B only new bone grew and attached to the surface of BCBB. BCBB degraded more with the time and more new bone formed. The histological evaluation showed that the bone forming area in group A was more than that in group B, and there was significant difference between group A and group B (P lt; 0.05). Conclusion The osteogenesis of BCBB with tissue engineering was superior to only BCBB, BCBB could be used as a scaffold of bone tissue engineering.
Human lymphocyte function-associated antigen 3 (hLFA3) has been identified as an important T cell accessory molecule. Rhesus monkeys (Macaca mulatta) have been widely used as animal models for human immune disorders. Due to the species-specificity of immune system, it is necessary to study M. mulatta LFA3 (mmLFA3). In this study, the gene encoding mmLFA3 CD2-binding domain (mmLFA3Sh) was amplified by polymerase chain reaction (PCR) and genetically fused to human IgG1 Fc fragment in pPIC9K to construct the expression plasmid pPIC9K-mmLFA3Sh-Ig. Approximately 3-4 mg mmLFA3Sh-Ig protein was recovered from 1 L of inductive media, and mmLFA3Sh-Ig produced by the P. pastoris can bind to the CD2 positive cells, and suppress the monkey and human lymphocytes proliferation induced by Con A and alloantigen in a dose-dependent manner. These results suggested that mmLFA3Sh-Ig might be used as a novel tool for pathogenesis and experimental immunotherapy of Rhesus monkey immune disorders.
Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) might be developed as a novel anti-tumor drug due to its selective cytotoxicity in tumor cells. The predicted Macaca mulatta TRAIL (mmTRAIL) is highly homologous to hTRAIL in nucleotide acid as well as amino acid sequence, suggesting that mmTRAIL might induce apoptosis of human cancer cells. However, the cytotoxicity of mmTRAIL in human cancer cells has not been investigated. In this paper, it is reported that the gene encoding mmTRAIL has been cloned by using reverse-transcriptase polymerase chain reaction (RT-PCR) from monkey peripheral blood mononuclear cells (PBMCs) in our laboratory. Subsequently, an expression plasmid was constructed by inserting mmTRAIL gene into pQE30 plasmid. After induction by addition of Isopropylβ-D-1-Thiogalactopyranoside (IPTG), mmTRAIL was expressed. MmTRAIL was recovered from supernatant of sonicated bacteria by Ni-NTA agarose affinity chromatography. SDS-PAGE and gel filtration chromatography demonstrated that mmTRAIL forms trimer in solution. In vitro assays indicated that mmTRAIL was cytotoxic to human COLO205 tumor cells but not to normal cells at low concentration of nanomole. In addition, antitumor effect of mmTRAIL was evaluated in mice bearing human COLO205 tumor xenografts. Intratumorally injected mmTRAIL significantly inhibited growth of tumor grafts. These results suggested that mmTRAIL was valuable as candidate drug for cancer-targeted therapy.
ObjectiveTo investigate the feasibility of electroencephalography (EEG) power spectrum analysis monitoring noninvasive intracranial pressure (ICP). MethodsBetween September 2008 and May 2009, the EEG signals were recorded in 62 patients (70 cases/times) with central nervous system (CNS). By using self-designed software, EEG power spectrum analysis was conducted and pressure index (PI) was calculated automatically. ICP was measured by lumbar puncture (LP). ResultsThe mean ICP was (239.74±116.25) mm H2O (70-500 mm H2O, 1 mm H2O=0.009 8 kPa), and 52.9% of patients had increased ICP. The mean PI was 0.29±0.20 (0.02-0.85). The Spearman rank test showed that there was a significant negative correlation between PI and ICP (rs=-0.849, P<0.01). The data from the patients with diffuse lesions of CNS and focal lesions were analyzed separately; the results showed there were significant negative correlations between PI and ICP in both groups (rs=-0.815, -0.912; P<0.01). ConclusionThe PI obtained from EEG analysis is correlated with ICP. Analysis of specific parameters from EEG power spectrum might reflect the ICP. Further research should be carried out.
This paper describes a simulation of microwave brain imaging for the detection of hemorrhagic stroke. Firstly, in the research process, the formula of DebyeⅡwas used to study tissues of brain and blood clot so that microwave frequency band was confirmed for imaging. Then a model with electromagnetic characteristics of brain was built on this basis. In addition, an ultra-wideband (UWB) Vivaldi antenna is designed to use for transmitting and receiving microwave signals of widths 1.7 GHz to 4 GHz. Microwave signals were transmitted and received when the antenna revolved around the brain model. Symmetric position de-noising method was used to eliminate the strong background noise signals, and finally confocal imaging method was applied to get brain imaging. Blood clot was distinguished clearly from result of imaging and position error was less than 1 cm.
ObjectiveTo understand the obstacles in the practice of pulmonary rehabilitation between doctors and patients.MethodsMedical staff and patients with chronic obstructive pulmonary disease (COPD) in public hospitals in this region were randomly sampled, and a questionnaire survey was conducted on possible obstacles to the practice of pulmonary rehabilitation.ResultsTotal of 265 medical staff and 120 COPD patients were recruited in this survey. The obstacles of pulmonary rehabilitation practice of medical staff in clinical work are poor cooperation of patients and their families (84.2%), medical staff’s insufficient awareness of pulmonary rehabilitation (82.3%), and lack of practice and guidelines (78.9%), lack of objective conditions such as site, equipment and equipment (75.1%), lack of multidisciplinary teams (74.3%), pulmonary rehabilitation has fallen by the wayside (73.6%) etc. The main obstacles for COPD patients in pulmonary rehabilitation are lack of access to relevant knowledge (52.4%), insufficient knowledge (36.5%), inconvenient transportation, economic problems and other objective conditions (33.3%). After multiple correspondence analysis, there are differences in the degree of correlation between obstacle factors and groups with different characteristics.ConclusionsThere are many factors hindering the development of pulmonary rehabilitation and there are certain differences among different populations, but the lack of understanding of pulmonary rehabilitation between doctors and patients is the primary problem. It is necessary to improve the cognition of both doctors and patients on pulmonary rehabilitation, and then to solve the obstacles in the implementation of pulmonary rehabilitation.
ObjectiveTo prepare platelet-derived growth factor receptor β (PDGFRβ)-targeted near-infrared molecular probe and evaluate its potential in optical molecular imaging of lung cancer.MethodsPDGFRβ-specific affibody Z-tri was recombinantly expressed in Escherichia coli (E. coli) and purified using affinity chromatography. In vitro cell-binding of Z-tri was analyzed by flow cytometry. Cellular distribution of Z-tri in tumor grafts was determined by protein-tracing. The molecular probe CF750-Z-tri was prepared by conjugating near-infrared fluorescent dye CF750 to Z-tri. The optical images of xenografts of lung cancer were obtained by using CF750-Z-tri combined with optical imaging system.ResultsPDGFRβ-specific affibody Z-tri was highly expressed in E. coli and purified to homogeneity. Z-tri could bind PDGFRβ-positive cells but not PDGFRβ-negative cells cultured in vitro. In the tumor xenografts of human lung cancer, intravenously injected Z-tri was predominantly distributed on cells overexpressing PDGFRβ. The near infrared fluorescent dye CF750 was efficiently conjugated to Z-tri. Optical images with high contrast of lung cancer xenografts were produced by using the near-infrared fluorescent probe CF750-Z-tri combined with optical imaging system.ConclusionThe near-infrared fluorescent probe CF750-Z-tri can be used for optical imaging of human lung cancer, which takes great potential in optical imaging-guided surgery of lung cancer.
Objective To explore the role of cyclin B1 (CCNB1), cyclin B2 (CCNB2) and cyclin dependent kinase 1 (CDK1) in lung adenocarcinoma (LUAD) using bioinformatic data. Methods First, RNA expression data were downloaded from two datasets in Gene Expression Omnibus (GEO), and DESeq2 software was used to identify deferentially expressed genes (DEGs). Subsequent analyses were conducted based on the results of these DEGs: protein-protein interaction (PPI) network was constructed with STRING database; the modules in PPI network were analyzed by Molecular Complex Detection software, and the most significant modules were selected, the genes included in these modules were the hub genes; high-throughput RNA sequencing data from other databases were used to verify the expression of these hub genes to confirm whether they were DEGs; survival curve analyses of the confirmed DEGs were conducted to select genes that had significant influence on the survival of LUAD; the expression of these hub genes in different stages of LUAD were also analyzed. Then, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis was performed for these selected hub genes using KOBAS database. MuTarget tool was used to analyze the correlations between the expression of these selected hub genes and gene mutation status in LUAD. The potential value of these hub genes in the treatment of LUAD was explored based on the drug information in GDSC database. Finally, immunohistochemical data from Human Protein Atlas (HPA) database were used to verify the expression of these hub genes in LUAD again. Results According to the expression data in GEO, 594 up-regulated genes and 651 down-regulated genes were identified (P<0.05), among which 30 hub genes were selected for subsequent analyses. The RNA high-throughput sequencing data of other databases verified that 18 genes were DEGs, among which 8 hub genes had significant impact on disease-free survival in LUAD (P<0.05). Moreover, the 8 genes were differentially expressed in different stages of LUAD, which were higher in the middle and late stage of LUAD. Among the 8 genes. CCNB1, CCNB2 and CDK1 were significantly enriched in the cell cycle pathway. The expression of CCNB1, CCNB2 and CDK1 in LUAD was closely related to the TP53 mutation status. In addition, CDK1 was associated with four drugs, revealing the potential value of CDK1 in the treatment of LUAD. Finally, immunohistochemical data from HPA database verified that CCNB1, CCNB2 and CDK1 were highly expressed in LUAD in the protein level. Conclusion Overexpression of CCNB1, CCNB2 and CDK1 are associated with poor prognosis of LUAD, indicating that the three genes may be prognostic biomarkers of LUAD and CDK1 is a potential therapeutic target for LUAD.