To improve prokaryotic expression level of Lagurus lagurus zona pellucida 3(LZP3) gene.The rare codons of LZP3 gene fragment were turned into the most frequently used codons in E. coli by overlap PCR. LZP3 gene and mutated LZP3 (LZP3m) gene were cloned into expression vector pGEX4T-1 respectively, and transformed into E. coli BL21(DE3).The expression level of LZP3m was notably higher than that of LZP3.The expression of LZP3 gene in E. coli could be successfully improved by using codon optimization.
ABSTRACT Cells employ multiple systems to maintain cellular integrity, including mechanosensitive (MS) ion channels and the cell wall integrity (CWI) pathway. Here, we use pollen as a model system to ask how these different mechanisms are interconnected at the cellular level. MscS-Like (MSL)8 is an MS channel required to protect Arabidopsis thaliana pollen from osmotic challenges during in vitro rehydration, germination and tube growth. New CRISPR/Cas9 and artificial microRNA-generated msl8 alleles produced unexpected pollen phenotypes, including the ability to germinate a tube after bursting, dramatic defects in cell wall structure and disorganized callose deposition at the germination site. We document complex genetic interactions between MSL8 and two previously established components of the CWI pathway, MARIS , and ANXUR1/2 . Overexpression of MARIS R240C -FP suppressed the bursting, germination, and callose deposition phenotypes of msl8 mutant pollen. Null msl8 alleles suppressed the internalized callose structures observed in MARIS R240C -FP lines. Similarly, MSL8-YFP overexpression suppressed bursting in the anxur1/2 mutant background, while anxur1/2 alleles reduced the strong rings of callose around ungerminated pollen grains in MSL8-YFP over-expressors. These data show that MS ion channels modulate callose deposition in pollen and provides evidence that cell wall and membrane surveillance systems coordinate in a complex manner to maintain cell integrity.
The large incidence of bone defects in clinical practice increases not only the demand for advanced bone transplantation techniques but also the development of bone substitute materials. A variety of emerging bone tissue engineering materials with osteogenic induction ability are promising strategies for the design of bone substitutes. MicroRNAs (miRNAs) are a class of non-coding RNAs that regulate intracellular protein expression by targeting the non-coding region of mRNA3′-UTR to play an important role in osteogenic differentiation. Several miRNA preparations have been used to promote the osteogenic differentiation of stem cells. Therefore, multiple functional bone tissue engineering materials using miRNA as an osteogenic factor have been developed and confirmed to have critical efficacy in promoting bone repair. In this review, osteogenic intracellular signaling pathways mediated by miRNAs are introduced in detail to provide a clear understanding for future clinical treatment. We summarized the biomaterials loaded with exogenous cells engineered by miRNAs and biomaterials directly carrying miRNAs acting on endogenous stem cells and discussed their advantages and disadvantages, providing a feasible method for promoting bone regeneration. Finally, we summarized the current research deficiencies and future research directions of the miRNA-functionalized scaffold. This review provides a summary of a variety of advanced miRNA delivery system design strategies that enhance bone regeneration.
Objective: To evaluate and comprehensively analyze the clinical efficacy of recombinant human endostatin combined with Iressa targeted therapy in patients with pleural metastasis of lung adenocarcinoma. Methods: The interval of the selected study period span was from January 2017 to April 2021. The sample source of the study was 42 patients with lung adenocarcinoma admitted to hospital. The random number table method was used for study grouping, and they were further divided into study groups (n = 21, 14 cases with pleural metastasis) and control group (n=21, 13 cases with pleural metastasis), all patients received systemic chemotherapy with pemetrexed and cisplatin. Patients with pleural metastases in the control group were injected with 60 mg cisplatin into the thoracic cavity. Patients in the study group were treated with Iressa (gefitinib) targeted therapy if genetic testing showed epidermal growth factor receptor (EGRF) mutations, and patients with pleural metastases were treated with pleural metastasis with Endo (recombinant human endostatin YH-16) to control pleural effusion. Two sets of related indicators were compared and analyzed. Results: Comparing the short-term disease control rate, treatment effectiveness and long-term survival rate between the two groups shows that the study group has more advantages (P<0.05). In the comparison between the two groups of serum markers and related indicators, the study group has more advantages (P<0.05), whereas in the comparison between the two groups in the incidence of adverse reactions, there is no significant difference (P>0.05). Based on statistics of the recurrence rate of pleural fluid in the two groups, the study group is significantly lower than the control group (P<0.05). Conclusion: Recombinant human endostatin combined with Iressa targeted therapy for patients with lung adenocarcinoma with pleural metastasis has significant short-term and long-term effects without serious adverse reactions. It can be fully promoted in medical institutions at all levels.
Background: Alzheimer’s disease (AD) occurs in the elderly and pre-elderly, characterized by decline of memory, cognitive dysfunction, impairment of learning capacity, and motor dysfunction. Recently a competitive endogenous RNA (ceRNA) network has been found to be related to AD progression, but there is still little understanding of the ceRNA regulatory network in AD. This study aims to explore the important regulatory mechanisms of ceRNA regulatory networks containing long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs) in AD. Methods: Data from the gene expression omnibus (GEO) database were used for the analysis. To study enrichment function for the upregulated and downregulated mRNAs, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using the Metascape database, respectively. Based on the STRING database and Cytoscape software 3.9.1, a protein-protein interaction (PPI) network was constructed. The hub genes in this network were identified utilizing the CytoHubba plugin in Cytoscape. The TargetScan, miRWalk, and miRDB were selected to calculate the regulatory interaction between miRNAs and the hub genes. LncRNAs were predicted using RNA22. Additionally, circRNA prediction was executed using the circBank database. Results: 711 downregulated and 670 upregulated overlapping mRNAs were identified between AD and control samples. 32 downregulated and 340 upregulated miRNAs were obtained from AD samples compared with control samples. 78 upregulated and 205 downregulated circRNAs were screened. 275 upregulated lncRNAs and 209 downregulated lncRNAs were found between AD samples and control samples. The PPI network constructed consists of 1016 nodes and 13,946 edges. Ten hub genes were selected to identify target miRNAs and ceRNAs. On the basis of the ceRNA hypothesis, a circRNA/lncRNA-miRNA-mRNA network was established. It included five lncRNAs (TRHDE-AS1, SNHG10, OIP5-AS, LINC00926 and LINC00662), 26 circRNAs, five miRNAs (hsa-miR-3158-3p, hsa-miR-4435, hsa-let-7d-3p, hsa-miR-330-5p and hsa-miR-3605-3p), and ten mRNAs (RPL11, RPL34, RPL21, RPL22, RPL6, RPL32, RPL24, RPL35, RPL31, and RPL35A). RPL35 and RPL35A were found to be significantly associated with AD pathology in tau and Aβ line AD models by the AlzData database. The study discovered the significance of several lncRNA-miRNA-mRNA axes and circRNA-miRNA-mRNA axes that included RPL35A and RPL35. Conclusions: ceRNAs were found to be important regulators in the development of AD and provide potential biological therapy targets for AD management.
Panax ginseng C.A.Meyer is one of the most valuable traditional Chinese medicines. In this study, the essential oil of ginseng leaves (EOGL), collected using hydrodistillation and analyzed by GC/MS, contained a complex mixture of aliphatic (69.0%), terpenoid (21.5%) and aromatic compounds (2.4%). Among 54 components identified, the major ones were palmitic acid (36.1%), β-farnesene (15.4%), linoleic acid (9.8%) and phytol (5.6%). In the cytotoxicity study, EOGL exhibited obvious cytotoxic activities against different cancer cell lines, including Hela, A549, ZR-75-1, HT-29, SGC7901 and B16 cells. Furthermore, Annexin V-FITC/PI staining assay indicated that EOGL can induce late apoptosis of ZR-75-1 cells, and the percentage of apoptotic cells increased in a concentration-dependent manner (0.9% to 5.6% and 67.4%). In addition to this, we also found that EOGL exhibited weak DPPH radical scavenging (12.0 ± 0.4 mg/mL) and ABTS radical scavenging activities (1.6 ± 0.1 mg/mL), and showed antibacterial activity against the Gram-positive bacteria, Staphylococcus aureus and Bacillus subtilis, and the Gram-negative bacterium, Escherichia coli. The data suggest that EOGL, which possesses important biological activities, especially significant anticancer activity, could be a potential medicinal resource.
An efficient NCC pincer Ni (II)‐catalyzed hydrophosphination of nitroalkenes with diphenylphosphine has been developed. Under the optimized conditions, both (hetero)aromatic and aliphatic nitroalkenes were well tolerated, irrespective of electronic effect, to provide the corresponding products in up to 99% yield.
Abstract: Background: Asthma is one of the most prevalent, complex, and severe respiratory disorders, with rising rates of occurrence and death in both children and adults. An alkaloid derived from plants, lycorine possesses several pharmacological effects, primarily anti-inflammatory and antioxidative properties. Materials and Methods: In the current study, the anti-asthmatic potential of lycorine against ovalbumin (OVA)-induced asthma mice model has been examined. The animals were categorized into five groups. The group I mice received saline while the group II mice received OVA. Groups III mice were provided with OVA + Lycorine in low dose whereas group IV mice were given OVA + Lycorine in high dose. Group V animals received OVA + Dexamethasone, which was used as the positive control. The bronchoalveolar lavage fluid isolated from the lungs was utilized to estimate various parameters such as total and differential immune cells (eosinophils, neutrophils, lymphocytes, macrophages), secreted inflammatory cytokines levels, and oxidative stress markers. The lungs were subjected to histological examination and respiratory mechanics examination. Results: Lycorine treatment significantly suppressed the total and differential immune cells, IgE, IL-4, IL-5, IL-13, and TNF-α cytokines, MDA, NO2 and NO3 levels and enhanced the IFN-γ cytokine level along with the SOD, CAT, and GSH enzyme activities. Further, the histopathological study revealed that lycorine was able to decrease the thickness of the columnar epithelial cells in the airways. Thus, lycorine suppressed the ROS generation that in turn reduced the inflammation in the airways of the lungs by reducing Th2 cytokines and augmenting the levels of Th1 cells. Conclusion: These findings indicate that lycorine was able to regulate the Th1/Th2 balance OVA-challenged asthmatic mice, signifying its protective impact that may be valuable in the management of allergic asthma. Keywords: Asthma, Ovalbumin, Lycorine, Inflammatory cytokines, Oxidative stress.
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