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.

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.

Mannan-modified solid lipid nanoparticles for targeted gene delivery to alveolar macrophages.

PURPOSE: Cationic solid lipid nanoparticles (SLN) have established themselves during the past decades. They can efficiently bind DNA directly via ionic interaction and mediate gene transfection. One major problem with SLN is the lack of cell-targeting ability. In the present study, a mannan-based PE-grafted ligand was synthesized and used for the surface modification of DNA-loaded cationic SLN to prepare Man-SLN-DNA. METHODS: For in vitro test, the cytotoxicity and transfection investigation was carried out on murine macrophage cell line RAW 264.7. For in vivo evaluation, Man-SLN-DNA was delivered into the lung of the rats, and the alveolar macrophages (AM) were isolated for the fluorescence determination of transfection efficiency. RESULTS: When compared with non-modified SLN-DNA and Lipofectamine 2000-DNA, Man-SLN-DNA produced the highest gene expressions, especially in vivo. CONCLUSION: These results demonstrated the active targeting ability of this kind of mannan-modified DNA-loaded vehicles, which may have great potential for targeted gene delivery.