A strategy for efficient gene delivery and expression was developed by encapsulating complexes of plasmid DNA and heparin-modified polyethylenimine into thermosensitive dextran–poly(ε-caprolactone)–2-hydroxylethylmethacrylate–poly(N-isopropylacrylamide) (Dex-PCL-HEMA-PNIPAAm) hydrogels. Thermosensitive hydrogels (THs) with different compositions were prepared and the TH with the highest content of PCL and Dex had the smallest pore size. Branched polyethylenimine (PEI) and heparin (Hep) could spontaneously form Hep/PEI complex particles in aqueous solution with the Hep content ranging from 20–50%. The Hep/PEI complex particles with 20% Hep (HP20), which were spherical in morphology, were well-dispersed and possessed a narrow size distribution with an average diameter of 209 nm. HP20/DNA complexes were then encapsulated in different THs. The release of HP20/DNA complexes from the hydrogels occurred mainly within the first 72 h in phosphate-buffered saline. HP20/DNA complexes used to transfect HeLa and 293T cells yielded luciferase expression levels which were 50–140-fold higher in serum-containing media, compared with PEI/DNA complexes in serum-free media. HP20/DNA complexes encapsulated in Dex-PCL-HEMA-PNIPAAm hydrogels not only exhibited appreciable transfection efficiency, but also provided a prolonged transgene expression, indicating that the combination of heparin-modified PEI and thermosensitive hydrogels has great potential for localized gene delivery.
Temperature- and pH-sensitive poly(N-isopropylacrylamide-co-acrylic acid-co-poly(ε-caprolactone)) (P(NIPAAm-co-AAc-co-PCL)) hydrogels were synthesized to immobilize plasmid DNA (pDNA) compacted with heparin-modified polyethyleneimine (Hep/PEI). The interior morphology of the prepared hydrogels observed by SEM showed macroporous structure clearly. It was found that the swollen hydrogels with low PCL content were able to immobilize efficiently plasmid DNA complexes. The de-swollen hydrogels resulted in a sustained release of the immobilized DNA complexes, and the released DNA complexes exhibited considerable gene expression activity with reduced cytotoxicity as compared with DNA complexes in the solution, indicating that the heparin-modified PEI/DNA-complex-immobilized environment-sensitive hydrogel has promising applications in substrate-mediated gene delivery.
It was controversial to operate on the primary site of breast cancer (BC) with bone metastasis only. We investigated the impact of surgery on BC patients with bone metastases via a SEER database retrospective analysis.
Presently, no study reported the function of cathepsin H (CTSH) in thyroid carcinoma (THCA). The aim of present study was to initially explore the factors affecting CTSH expression, and association between CTSH expression and survival rate in THCA.We explored mRNA expression of CTSH in normal and BRCA tissues, and evaluated prognostic impact of CTSH expression on the overall survival of THCA patients. Then, related factors influencing CTSH mRNA expression in THCA were analyzed. Functional enrichment analysis was performed to reveal the potential function of CTSH involved in THCA. We also constructed PPI network among co-expressed genes of CTSH to determine hub genes, followed by association analysis on hub genes with CTSH.(1) CTSH mRNA was highly expressed in THCA compared with normal group (P<0.001). High expression of CTSH was conducive to the overall survival of THCA patients (P=0.0027). CTSH was then determined as an independent prognostic factor in THCA (P=0.024). (2) The mRNA expression of CTSH was statistically related to patient's histological type, N stage, T stage, tumor stage and sample type (all P<0.001). CTSH copy number variation and methylation also influenced its mRNA expression (all P<0.001). (3) Pathway analysis indicated that CTSH mainly participated in cancer-related pathways, such as hedgehog signaling pathway, cytokine-cytokine receptor interaction and JAK-STAT signaling pathway (all P<0.05). (4) The top 10 co-expressed genes in whole PPI network showed significant correlation with CTSH expression (all P<0.001).CTSH higher expression was observed in THCA, which caused a good prognosis of patients. CTSH expression might be regulated by multiple factors including clinical characteristic, methylation, copy number and other genes. This study demonstrated the clinical significance of CTSH in THCA, as well as revealed the potential pathway associated with CTSH involved in thyroid cancer.
This meta-analysis aimed to evaluate the diagnostic accuracy of touch imprint cytology (TIC) for sentinel lymph node (SLN) metastases of patients with clinical node-negative early breast cancer. The PubMed, Web of Science, Embase, and the Cochrane Library databases were meticulously searched to retrieve literature published from January 2005 to September 2022 by two independent reviewers. The meta-analysis was performed using STATA16.0, Meta-Disc 1.4, and RevMan 5.4.9. According to the inclusion criteria, 4,073 patients from 13 studies were included in this meta-analysis. The pooled sensitivity and specificity of TIC for detecting SLN metastases were 0.77 (95% CI 0.66-0.85) and 0.99 (95% CI 0.97-1.00), respectively. The pooled positive likelihood ratio and negative likelihood ratio were 76.15 (95% CI 29.16-198.84) and 0.23 (95% CI 0.15-0.36), respectively. The pooled diagnostic odds ratio was 326.82 (95% CI 132.76-804.56) and the area under the sROC curve was 0.97 (95% CI 0.95-0.98), respectively. This meta-analysis revealed that TIC with high sensitivity and specificity is a feasibility and accuracy diagnosis technique for intraoperative detection of SLN metastases in breast cancer.
RNAi is regarded as a promising technology for pest control. However, not all insects are sensitive to RNAi. Studies have confirmed that insect dsRNases are one of key factors affecting RNAi efficiency. In the current study, we identified four genes coding for dsRNases from the Spodoptera frugiperda genome. Spatial and temporal expression analysis showed that those dsRNases were highly expressed in the midgut and old larvae. Then a delivery method was applied for inducing efficient RNAi based on dsRNA encapsulated by liposome. Furthermore, we assessed degradation efficiency by incubation with dsRNA with gut juice or hemocoel to characterize potential roles of different SfdsRNases after suppression of SfdsRNase. The result showed that interferenced with any sfdsRNase reduced the degradation of exogenous dsRNA in midgut, interfered with sfdsRNase1 and sfdsRNase3 slowed down the degradation of exogenous dsRNA in hemolymph. Our data suggest the evolutionary expansion and multiple high activity dsRNase genes would take part in the RNAi obstinate in S. frugiperda, besides we also provide an efficient RNAi method for better use of RNAi in S. frugiperda.
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