Prevalence and risk factors of early postoperative seizures in patients with glioma: A protocol for meta-analysis and systematic review.

INTRODUCTION: Early postoperative seizures has been the most common clinical expression in gliomas; however, the incidence and risk factors for early postoperative seizures in gliomas are more controversial. This protocol describes a systematic review and meta-analysis to clarify the prevalence and risk factors of early postoperative seizures in patients with glioma. METHODS AND ANALYSIS: Searches will be conducted on CNKI, WanFang, VIP, PubMed, Embase, Cochrane Library databases and Web of Science for the period from database inception to December 31st, 2023. Case-control and cohort studies of the incidence and risk factors for early postoperative seizures in all gliomas will be included. The primary outcome will be incidence, risk factors. Newcastle-Ottawa Scale was used for quality evaluation. Review of article screening, extracting data and risk of bias assessment will be repeated by two independent reviewers. RESULT: This study will provide evidence for the risk factors and incidence of early postoperative seizures in patients with glioma. CONCLUSION: Our study will provide evidence for the prevention of early postoperative seizures in glioma patients. TRAIL REGISTRATION: This protocol was registered in PROSPERO and registration number is CRD42023415658.

Upregulation of HLA-II related to LAG-3+CD4+ T cell infiltration is associated with patient outcome in human glioblastoma.

Glioblastoma (GBM) is the most common malignant diffuse glioma of the brain. Although immunotherapy with immune checkpoint inhibitors (ICIs), such as programmed cell death protein (PD)-1/PD ligand-1 inhibitors, has revolutionized the treatment of several cancers, the clinical benefit in GBM patients has been limited. Lymphocyte-activation gene 3 (LAG-3) binding to human leukocyte antigen-II (HLA-II) plays an essential role in triggering CD4+ T cell exhaustion and could interfere with the efficiency of anti-PD-1 treatment; however, the value of LAG-3-HLA-II interactions in ICI immunotherapy for GBM patients has not yet been analyzed. Therefore, we aimed to investigate the expression and regulation of HLA-II in human GBM samples and the correlation with LAG-3+CD4+ T cell infiltration. Human leukocyte antigen-II was highly expressed in GBM and correlated with increased LAG-3+CD4+ T cell infiltration in the stroma. Additionally, HLA-IIHighLAG-3High was associated with worse patient survival. Increased interleukin-10 (IL-10) expression was observed in GBM, which was correlated with high levels of HLA-II and LAG-3+ T cell infiltration in stroma. HLA-IIHighIL-10High GBM associated with LAG-3+ T cells infiltration synergistically showed shorter overall survival in patients. Combined anti-LAG-3 and anti-IL-10 treatment inhibited tumor growth in a mouse brain GL261 tumor model. In vitro, CD68+ macrophages upregulated HLA-II expression in GBM cells through tumor necrosis factor-α (TNF-α). Blocking TNF-α-dependent inflammation inhibited tumor growth in a mouse GBM model. In summary, T cell-tumor cell interactions, such as LAG-3-HLA-II, could confer an immunosuppressive environment in human GBM, leading to poor prognosis in patients. Therefore, targeting the LAG-3-HLA-II interaction could be beneficial in ICI immunotherapy to improve the clinical outcome of GBM patients.

[Retrospective clinical study of hip replacement in the treatment of traumatic arthritis secondary to acetabular fracture].

OBJECTIVE: To investigate the clinical effect of total hip replacement (THA) in the treatment of traumatic arthritis secondary to acetabular fracture. METHODS: From October 2019 to June 2022, 15 patients with secondary traumatic arthritis of acetabulum fracture were treated with THA. There were 8 males and 7 females, aged from 40 to 76 years old with an average of (59.20±9.46) years old. Prosthesis loosening, dislocation of hip joint, range of motion of hip joint, nerve injury and other conditions were recorded before and after surgery. Harris score, visual analogue scale (VAS) and imaging were used to evaluate hip joint function and surgical effect. RESULTS: Follow-up time ranged 6 to 39 months with an average of (18.33±9.27) months. All the 15 patients successfully completed the operation, no nerve and blood vessel injury during the operation, postoperative wound healing was stageⅠ, no infection, one case of acetabular side prosthesis loosening at half a year after operation, and recovered well after revision surgery, one case of hip dislocation was cured after open reduction treatment, no adverse consequences. Harris score at the last postoperative follow-up was (88.60±4.01) points, compared with the preoperative (47.20±11.77) points, the difference was statistically significant (P<0.05), and VAS at the lateat postoperative follow-up was 1 (1) points, compared with the preoperative 8 (2) points, the difference was statistically significant (P<0.05). At the last follow-up, the pain symptoms were relieved or disappeared, and the joint function was satisfactory. The imaging data of the latest follow-up showed joint was well pseudoradiated, no abnormal ossification occurred, and the prosthesis was not loose. CONCLUSION: THA is effective in the treatment of traumatic arthritis secondary to acetabular fracture and can effectively improve the quality of life of patients. Preoperative comprehensive evaluation and bone defect evaluation of patients, and intraoperative management of acetabulum, femur, internal fixation and bone defect are key factors for the success of surgery.

Prognostic marker CXCL5 in glioblastoma polyformis and its mechanism of immune invasion.

Glioblastoma multiforme (GBM) is the most aggressive brain cancer with a poor prognosis. Therefore, the correlative molecular markers and molecular mechanisms should be explored to assess the occurrence and treatment of glioma.WB and qPCR assays were used to detect the expression of CXCL5 in human GBM tissues. The relationship between CXCL5 expression and clinicopathological features was evaluated using logistic regression analysis, Wilcoxon symbolic rank test, and Kruskal-Wallis test. Univariate, multivariate Cox regression and Kaplan-Meier methods were used to assess CXCL5 and other prognostic factors of GBM. Gene set enrichment analysis (GSEA) was used to identify pathways associated with CXCL5. The correlation between CXCL5 and tumor immunoinfiltration was investigated using single sample gene set enrichment analysis (ssGSEA) of TCGA data. Cell experiments and mouse subcutaneous transplanted tumor models were used to evaluate the role of CXCL5 in GBM. WB, qPCR, immunofluorescence, and immunohistochemical assays showed that CXCL5 expression was increased in human GBM tissues. Furthermore, high CXCL5 expression was closely related to poor disease-specific survival and overall survival of GBM patients. The ssGSEA suggested that CXCL5 is closely related to the cell cycle and immune response through PPAR signaling pathway. GSEA also showed that CXCL5 expression was positively correlated with macrophage cell infiltration level and negatively correlated with cytotoxic cell infiltration level. CXCL5 may be associated with the prognosis and immunoinfiltration of GBM.

Modeling and analyzing the action process of monoamine hormones in depression: a Petri nets-based intelligent approach.

In contemporary society, the incidence of depression is increasing significantly around the world. At present, most of the treatment methods for depression are psychological counseling and drug therapy. However, this approach does not allow patients to visualize the logic of hormones at the pathological level. In order to better apply intelligence computing methods to the medical field, and to more easily analyze the relationship between norepinephrine and dopamine in depression, it is necessary to build an interpretable graphical model to analyze this relationship which is of great significance to help discover new treatment ideas and potential drug targets. Petri net (PN) is a mathematical and graphic tool used to simulate and study complex system processes. This article utilizes PN to study the relationship between norepinephrine and dopamine in depression. We use PN to model the relationship between the norepinephrine and dopamine, and then use the invariant method of PN to verify and analyze it. The mathematical model proposed in this article can explain the complex pathogenesis of depression and visualize the process of intracellular hormone-induced state changes. Finally, the experiment result suggests that our method provides some possible research directions and approaches for the development of antidepressant drugs.

Tracing COVID-19 drugs in the environment: Are we focusing on the right environmental compartment?

The Coronavirus Disease 2019 (COVID-19) pandemic led to over 770 million confirmed cases, straining public healthcare systems and necessitating extensive and prolonged use of synthetic chemical drugs around the globe for medical treatment and symptom relief. Concerns have arisen regarding the massive release of active pharmaceutical ingredients (APIs) and their metabolites into the environment, particularly through domestic sewage. While discussions surrounding this issue have primarily centered on their discharge into aquatic environments, particularly through treated effluent from municipal wastewater treatment plants (WWTPs), one often overlooked aspect is the terrestrial environment as a significant receptor of pharmaceutical-laden waste. This occurs through the disposal of sewage sludge, for instance, by applying biosolids to land or non-compliant disposal of sewage sludge, in addition to the routine disposal of expired and unused medications in municipal solid wastes. In this article, we surveyed sixteen approved pharmaceuticals for treating COVID-19 and bacterial co-infections, along with their primary metabolites. For this, we delved into their physiochemical properties, ecological toxicities, environmental persistence, and fate within municipal WWTPs. Emphasis was given on lipophilic substances with log Kow >3.0, which are more likely to be found in sewage sludge at significant factions (25.2%-75.0%) of their inputs in raw sewage and subsequently enter the terrestrial environment through land application of biosolids, e.g., 43% in the United States and as high as 96% in Ireland or non-compliant practices of sewage sludge disposal in developing communities, such as open dumping and land application without prior anaerobic digestion. The available evidence underscores the importance of adequately treating and disposing of sewage sludge before its final disposal or land application in an epidemic or pandemic scenario, as mismanaged sewage sludge could be a significant vector for releasing pharmaceutical compounds and their metabolites into the terrestrial environment.

Monocular 3D Human Pose Markerless Systems for Gait Assessment.

Gait analysis plays an important role in the fields of healthcare and sports sciences. Conventional gait analysis relies on costly equipment such as optical motion capture cameras and wearable sensors, some of which require trained assessors for data collection and processing. With the recent developments in computer vision and deep neural networks, using monocular RGB cameras for 3D human pose estimation has shown tremendous promise as a cost-effective and efficient solution for clinical gait analysis. In this paper, a markerless human pose technique is developed using motion captured by a consumer monocular camera (800 × 600 pixels and 30 FPS) for clinical gait analysis. The experimental results have shown that the proposed post-processing algorithm significantly improved the original human pose detection model (BlazePose)'s prediction performance compared to the gold-standard gait signals by 10.7% using the MoVi dataset. In addition, the predicted T2 score has an excellent correlation with ground truth (r = 0.99 and y = 0.94x + 0.01 regression line), which supports that our approach can be a potential alternative to the conventional marker-based solution to assist the clinical gait assessment.

Mental Health Study from the Perspective of Behavior Preference under the Environment of Internet Media: Based on China's Data Analysis.

To prevent the mental health problems of college preference students effectively, this paper analyzes the influence of college students' internet media use behaviors on their mental health in the information age. We make an empirical study on the above problems by using metrology models under the condition of controlling individual factors. The result shows that the mental health of college students is significantly affected by different internet media use behaviors and shows obvious heterogeneity. Preference for games and soap operas or films has a significant negative relationship with the mental health of college students, while the preference for science, education programs, and the preference to obtain current political news comments through official media have a positive impact on the psychological security of college students. Meanwhile, the mental health of college students is also significantly influenced by individual factors. The education level, family income, and social satisfaction show a significant positive correlation with the mental health of college students.

Dynamical Behaviors of a Translating Liquid Crystal Elastomer Fiber in a Linear Temperature Field.

Liquid crystal elastomer (LCE) fiber with a fixed end in an inhomogeneous temperature field is capable of self-oscillating because of coupling between heat transfer and deformation, and the dynamics of a translating LCE fiber in an inhomogeneous temperature field are worth investigating to widen its applications. In this paper, we propose a theoretic constitutive model and the asymptotic relationship of a LCE fiber translating in a linear temperature field and investigate the dynamical behaviors of a corresponding fiber-mass system. In the three cases of the frame at rest, uniform, and accelerating translation, the fiber-mass system can still self-oscillate, which is determined by the combination of the heat-transfer characteristic time, the temperature gradient, and the thermal expansion coefficient. The self-oscillation is maintained by the energy input from the ambient linear temperature field to compensate for damping dissipation. Meanwhile, the amplitude and frequency of the self-oscillation are not affected by the translating frame for the three cases. Compared with the cases of the frame at rest, the translating frame can change the equilibrium position of the self-oscillation. The results are expected to provide some useful recommendations for the design and motion control in the fields of micro-robots, energy harvesters, and clinical surgical scenarios.

The complete chloroplast genome of Arisaema bockii Engler and its phylogenetic analysis in the family Araceae.

Arisaema bockii Engler is a perennial herbaceous medicinal plant, which is widely distributed in many provinces in China such as Anhui, Jiangsu, and Zhejiang. In this study, the complete chloroplast genome sequence of A. bockii was assembled and characterized based on high-throughput sequencing data. The total length of chloroplast genome was 175,537 bp, including large single-copy (LSC) and small single-copy (SSC) regions of 98,870 bp and 23,345 bp, respectively, which were separated by a pair of 27,161 bp inverted repeat (IR) regions. The genome contained 129 genes, including 84 protein-coding genes, 36 tRNA genes, 8 rRNA genes, and one pseudogene. The overall GC content of the genome was 33.6%. A phylogenetic tree reconstructed by 30 chloroplast genomes revealed that A. bockii was mostly related to the same genus species A. ringens, A. franchetianum and A. erubescens. The work reported the first complete chloroplast genome of A. bockii, which may provide some useful information to the evolution of the family Araceae.

Sequencing and phylogenetic analysis of the complete chloroplast genome of Arisaema heterophyllum Blume.

Arisaema heterophyllum Blume is a perennial medicinal herb widely distributed in China, Korea and Japan. In this study, the complete chloroplast genome sequence of A. heterophyllum was assembled and characterized based on high-throughput sequencing data. The whole chloroplast genome is 170,610 bp in length and contains 95,485 bp large single-copy (LSC) and 22,605 bp small single-copy (SSC) regions separated by a pair of 26,260 bp inverted repeat (IR) regions. It contained a total of 129 genes, including 84 protein-coding genes, 37 tRNA genes, and 8 rRNA genes, with an overall GC content of 34.5%. A phylogenetic tree reconstructed by 30 chloroplast genomes reveals that A. heterophyllum is mostly related to the same genus A. ringens, A. franchetianum and A. erubescens. The complete chloroplast genome of A. heterophyllum was the firstly reported and deposited at GenBank under accession number MZ424448.

The complete chloroplast genome of wild Magnolia officinalis Rehd. et Wils.

Magnolia officinalis Rehd. et Wils. is an important traditional Chinese herbal medicine and widely distributed in the south of Yangtze River in China. In this study, the complete chloroplast genome sequence of wild M. officinalis was assembled and characterized from high-throughput sequencing data. The chloroplast genome was 160,009 bp in length, consisting of large single-copy (LSC) and small single-copy (SSC) regions of 88,134 bp and 18,739 bp, respectively, which were separated by a pair of 26,568 bp inverted repeat (IR) regions. The genome is predicted to contain 132 genes, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The overall GC content of the genome is 39.3%. A phylogenetic tree reconstructed by 86 chloroplast genomes reveals that M. officinalis is mostly related to cultivated M. officinalis, M. obovata and M. tripetala.

De novo assembly and Transcriptome Analysis of the Momordica charantia Seedlings Responding to methyl jasmonate using 454 pyrosequencing.

Momordica charantia, a medicinal and edible species of the Cucurbitaceae family, has been widely used as a vegetable around the world. Hundreds of pharmacological compounds isolated from the M. charantia have been reported. However, the mechanism of action of the secondary metabolites has not been fully elucidated. In this study, 118,590 unigenes were gained by de novo assembly based on the raw data from high-throughput sequencing of mRNA (RNA-Sequencing) upon systemic analysis, among which, 51,860 (43.73%) could be annotated to the public sequence databases such as Nr, GO, Swiss-Prot, KEGG and KOG. The transcriptomic changes of M. charantia seedlings treated with or without methyl jasmonate (MeJA) were analyzed to identify key genes involved in MeJA treatment. Additionally, 554 differentially expressed genes (DEGs), including 328 up-regulated ones and 226 down-regulated genes, have been identified. Most DEGs were associated with secondary metabolism and stress responses. Meanwhile, six DEGs were further confirmed by quantitative real-time RT-PCR (qRT-PCR) analysis, resulting in similar expression patterns as compared to those of RNA-Sequencing. Nine significantly enriched pathways including 11 DEGs were identified to be possibly involved in the MeJA-responsive biosynthesis of secondary metabolites based on the transcriptome sequencing analysis. Among them, 4 DEGs, encoding two peroxidases, one cinnamyl alcohol dehydrogenase and one hypothetical protein Csa, might play important roles in the process of phenylpropanoid biosynthesis. In addition, 9 transcription factors (TFs) were also detected as DEGs from 1899 unigenes. Most of them up-regulated by MeJA treatment might be potentially involved in regulating secondary metabolites biosynthesis. This work is the first research on the large-scale assessment of M. charantia transcriptomic resources and the analysis of DEGs and TFs in secondary metabolites biosynthesis of M. charantia seedings treated with or without MeJA, which will be conducive to the further applications of M. charantia.

Study on the efficient removal of azo dyes by heterogeneous photo-Fenton process with 3D flower-like layered double hydroxide.

As organic dyes are the main pollutants in water pollution, seeking effective removal solutions is urgent for humans and the environment. A novel environmentally friendly three-dimensional CoFe-LDHs (3D CoFe-LDHs) catalyst was synthesized by one-step hydrothermal method. Scanning electron microscopy, energy dispersive spectroscopy, Fourier transform infrared spectra, X-ray diffraction, X-ray photoelectron spectroscopy, Brunauer-Emmett-Teller technique as well as UV-Vis diffuse reflectance spectra were used to characterize the prepared samples. The experimental results revealed that 3D CoFe-LDHs exhibited a rapid decolorization of methyl orange and Rhodamine B by heterogeneous photo-Fenton process after reaching the adsorption equilibrium, and the final decolorization efficiency reached 91.18% and 93.56%, respectively. On the contrary, the decolorizing effect of 3D CoFe-LDHs on neutral blue was relatively weak. The initial concentrations of azo dyes, pH and H2O2 concentration affected the decolorization of dyes and the catalyst maintained excellent reusability and stability after reuse over five cycles. The quenching experiments found that •OH, •O2 - and h+ were the main active substances and reaction mechanisms were further proposed. The study suggests that the synergistic effect of photocatalysis and Fenton oxidation process significantly improved the removal of azo dyes and the synthesized catalyst had potentially promising applications for difficult-to-biodegrade organic pollutants in wastewater.

Non-Genomic Action of Androgens is Mediated by Rapid Phosphorylation and Regulation of Androgen Receptor Trafficking.

BACKGROUND: Testosterone is critical for maintaining spermatogenesis and male fertility. The accomplishment of these processes requires the synergistic actions of the classical and non-classical signaling pathways of androgens. METHODS: A murine testicular Sertoli cell line, TM4 cell was used to examine androgen actions in Sertoli cells. Western blot analysis and immunofluorescence assay were employed to study the testosterone-induced Androgen receptor (AR) translocation. Protein phosphorylation antibody array was applied to identify the phosphorylation sites under testosterone treatment, and these findings were verified by Western blot analysis. RESULTS: We found that a physiological dose of testosterone induced fast membrane association of AR. By using a phosphorylation antibody array, several phosphorylation sites, such as MEK1/2 (Ser217/221), Akt (Ser473), and Erk1/2 (Thr202/Tyr204) were rapidly phosphorylated within 5 min of testosterone treatment. Inhibition of the MEK and Akt signaling pathways prevented AR trafficking. Blocking of AR by flutamide eliminated the stimulation effect of testosterone on kinase phosphorylation. Testosterone induced kinase Src phosphorylation, and inhibition of Src restricted AR translocation to the membrane and the nucleus. CONCLUSION: Findings suggested that the membrane association of AR was mediated by the MEK and Akt phosphorylation signaling pathways, which resulted in Src activation and was initiated by testosterone binding to the membrane-localized AR. This study provides new insights into the testosterone signaling pathway in Sertoli cells, which mediate spermatogenesis. In addition, the study can be used in the diagnosis and treatment of male infertility caused by disorders in spermatogenesis.

Naturally occurring genetic mutations in the 5'-upstream regulatory region of bovine FSHB generate a novel cis-regulatory element that affects its expression.

We previously reported that numerous naturally occurring genetic mutations in the 5'-upstream regulatory region (5'-URR) of the bovine follicle-stimulating hormone beta-subunit gene (FSHB) were associated with reduced serum follicle-stimulating hormone (FSH) levels, poor-quality semen and low fertility in bulls. In addition, two different FSHB mRNA transcripts resulting from the linked mutations of genomic DNA were discovered in mutation-bearing bull pituitaries. Here, using electrophoretic mobility shift assay, we identified c.-1539_-1538delGGinsTTAACT mutations in the 5'-URR that generated a novel cis-regulatory element in bovine FSHB. Moreover, this novel element seemed to play a role in repressing FSHB transcription based on a promoter activity analysis in LßT2 gonadotrope cells. Quantitative assays of FSHB mRNA in the bovine pituitaries suggested that the levels of FSHB wild-type transcripts in the mutation-bearing bulls were significantly lower (P < 0.05) than in those of bulls without FSHB genetic mutations and that the levels of FSHB-mutated transcripts were significantly lower (P < 0.05) than that of wild-type transcripts in the mutation-bearing bulls. Altogether, our results suggest that decreased serum FSH levels and male fertility in bulls with the c.-1539_-1538delGGinsTTAACT mutation likely result from the alteration of cis-regulatory elements and induction of FSHB transcription.

Expression of microRNA­26b and identification of its target gene EphA2 in pituitary tissues in Yanbian cattle.

microRNAs (miRNAs/miRs) are a class of single­stranded non­coding RNA molecules of 19­24 nucleotides (nt) in length. They are widely expressed in animals, plants, bacteria and viruses. Via specific mRNA complementary pairing of target genes, miRNAs are able to regulate the expression of mRNA levels or inhibit protein translation following transcription. miRNA expression has a time­ and space specificity, and it is involved in cell proliferation and differentiation, apoptosis, development, tumor metastasis occurrence and other biological processes. miR­26b is an miRNA of 22 nt and is important in the regulation of cellular processes. With the advancement of molecular biology techniques in recent years, there have been extensive investigations into miR­26b. Numerous studies have observed that miR­26b is involved in early embryonic development, cell proliferation regulation, pituitary hormone secretion and other physiological activities. miRNAs are associated with the function of propagation. The present study used reverse transcription quantitative polymerase chain reaction to detect the relative expression levels of miR­26b in the pituitary tissue of Yanbian cattle at different developmental stages. The 2­∆∆Ct method was used to calculate the relative gene expression levels. The miRNA target gene database TargetScan and RNA22 were used for prediction of the miR­26b target gene and selective recognition was also performed. The results demonstrated that miR­26b is expressed in the pituitary tissues of Yanbian cattle at 6 and 24 months of age. The relative expression levels of miR­26b in the pituitary tissues of 24­month­old Yanbian cattle were 2.41 times that of those in the six­month­old Yanbian cattle, demonstrating significant differences in the relative expression (P<0.01). The relative expression of the candidate target genes, EphA2 and miR­26b, exhibited the opposite expression pattern. The relative expression levels in the pituitary tissues of six­month­old Yanbian cattle were 3.34 times that of those in 24­month­old Yanbian cattle (P<0.01). There are miR­26b binding sites in the 3'­untranslated region (3'­UTR) of EphA2 in bovine, human, murine and other mammalian mRNAs, suggesting that the EphA2 gene may be a target gene of miR­26b. The results of a Luciferase reporter system assay revealed that miR­26b is able to suppress EphA2 expression at the transcription level. Following the site­directed mutagenesis of plasmid EphA2 3'­UTR pmirGLO­MUT­ and miR­26b mimic­transfected HeLa cells, the dual­luciferase reporter gene assay revealed that there were three consecutive nucleotide mutations in the 3'­UTR, binding with the predicted seed region. This may have caused the miR­26b inhibition of luciferase activity to decrease from 60% in the wild­type to 26%, suggesting that miR­26b achieved its function via binding with the TACTTGAA sequence of the 3'­UTR in EphA2. In conclusion, the present study successfully assessed the expression pattern of miR­26b in the pituitary tissue of Yanbian cattle, and also confirmed that EphA2 was a target gene of miR­26b in Yanbian cattle in vitro. The present study provided the theoretical basis to further investigate the role of miR­26b in early embryonic development, pituitary hormone secretion and other reproductive functions.

Enhanced gene transfection efficiency in CD13-positive vascular endothelial cells with targeted poly(lactic acid)-poly(ethylene glycol) nanoparticles through caveolae-mediated endocytosis.

The efficient delivery of therapeutic gene into cells of interest is a critical challenge to broad application of non-viral vectors. The approach of introducing ligands that lead gene vectors to target caveolae-mediated endocytosis on nanoparticle surface might serve as a promising strategy for the effective gene transfection. Recently, in an attempt to enhance the possibility of caveolae-mediated endocytosis, we fabricated a peptide-targeted gene vector for highly efficient receptor-mediated intracellular delivery. Cyclic Asn-Gly-Arg (cNGR) peptide was used to target gene loaded poly(lactic acid)-poly(ethylene glycol) nanoparticles (PLA-PEG NPs) to HUVEC over-expressing CD13. Using 6-lauroxyhexyl lysinate (LHLN) as cationic surfactant, cNGR modified PLA-PEG NPs (cNGR-PEG-PLA NPs) were capable of complexing and compacting DNA into homogeneous small-sized complexes (<200nm) with positive charge (~10mV). Fortunately, the results of in vitro cellular uptake tests and mechanism studies were consistent with our original hypothesis. The cNGR peptide presented on nanoparticles' surface could specifically mediate the fast and efficient internalization of cNGR-PEG-PLA NPs into HUVEC. Moreover, free cNGR inhibited their intracellular uptake into HUVEC revealing the mechanism of receptor-mediated endocytosis. Furthermore, the inspiring results of the mechanism studies and transfection assays demonstrated that caveolae-mediated endocytosis was indeed mainly involved in the internalization of cNGR-PEG-PLA NPs into HUVEC and led to significant gene transfection efficiency in contrast with cNGR non-modified PLA-PEG NPs. Given such encouraging and favorable properties including biocompatibility, high transfer efficiency, low cytotoxicity, and fast uptake by nondestructive endocytic pathways, cNGR-PEG-PLA NPs could be a promising carrier for the intracellular delivery of therapeutic agents.

Novel cationic 6-lauroxyhexyl lysinate modified poly(lactic acid)-poly(ethylene glycol) nanoparticles enhance gene transfection.

The leading principle of non-viral delivery systems for gene therapy is to mediate high levels of gene expression with low cytotoxicity. Nowadays, biodegradable nanoparticles formulated with poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) were wildly developed. However, the relative lower gene transfection efficiency and higher cytotoxicity still remained critical problems. To address these limitations, PLA-PEG nanoparticles have been composited with other components in their formulation. Here, a novel cationic lipid, 6-lauroxyhexyl lysinate (LHLN), was fabricated onto PLA-PEG nanoparticles as a charge modifier to improve the transfection efficiency and cytotoxicity. The obtained cationic LHLN modified PLA-PEG nanoparticles (LHLN-PLA-PEG NPs) could condense pDNA thoroughly via electrostatic force, leading to the formation of the LHLN-PLA-PEG NPs/pDNA complexes (NPs/DNA complexes). The nanoparticles obtained have been characterized in relation to their physicochemical and biological properties, and the results are extremely promising in terms of low cell toxicity and high transfection efficiency. These results indicated that the novel cationic LHLN modified PLA-PEG nanoparticles could enhance gene transfection in vitro and hold the potential to be a promising non-viral nanodevice.