Immunisation Using Novel DNA Vaccine Encoding Virus Membrane Fusion Complex and Chemokine Genes Shows High Protection from HSV-2

Herpes simplex virus 1 and 2 infections cause high unmet disease burdens worldwide. Mainly HSV-2 causes persistent sexually transmitted disease, fatal neonatal disease and increased transmission of HIV/AIDS. Thus, there is an urgent requirement to develop effective vaccines. We developed nucleic acid vaccines encoding a novel virus entry complex stabilising cell membrane fusion, ‘virus-like membranes’, VLM. Two dose intramuscular immunisations using DNA expression plasmids in a guinea pig model gave 100% protection against acute disease and significantly reduced virus replication after virus intravaginal challenge. There was also reduced establishment of latency within the dorsal root ganglia and spinal cord, but recurrent disease and recurrent virus shedding remained. To increase cellular immunity and protect against recurrent disease, cDNA encoding an inhibitor of chemokine receptors on T regulatory cells was added and compared to chemokine CCL5 effects. Immunisation including this novel human chemokine gene, newly defined splice variant from an endogenous virus genome, ‘virokine immune therapeutic’, VIT, protected most guinea pigs from recurrent disease and reduced recurrent virus shedding distinct from a gD protein vaccine similar to that previously evaluated in clinical trials. All DNA vaccines induced significant neutralising antibodies and warrant evaluation for new therapeutic treatments.     

Do methylenetetrahydrofolate dehydrogenase,cyclohydrolase, and formyltetrahydrofolate synthetase 1 polymorphisms modify changes in intelligence of school-age children in areas of endemic fluorosis?

Background:Excessive exposure to fluoride can reduce intelligence. Methylenetetrahydrofolate dehydrogenase, cyclohydrolase, and formyltetrahydrofolate synthetase 1 (MTHFD1) polymorphisms have important roles in neurodevelopment. However, the association of MTHFD1 polymorphisms with children''s intelligence changes in endemic fluorosis areas has been rarely explored.Methods:A cross-sectional study was conducted in four randomly selected primary schools in Tongxu County, Henan Province, from April to May in 2017. A total of 694 children aged 8 to 12 years were included in the study with the recruitment by the cluster sampling method. Urinary fluoride (UF) and urinary creatinine were separately determined using the fluoride ion-selective electrode and creatinine assay kit. Children were classified as the high fluoride group and control group according to the median of urinary creatinine-adjusted urinary fluoride (UF_Cr) level. Four loci of MTHFD1 were genotyped, and the Combined Raven''s Test was used to evaluate children''s intelligence quotient (IQ). Generalized linear model and multinomial logistic regression model were performed to analyze the associations between children''s UF_Cr level, MTHFD1 polymorphisms, and intelligence. The general linear model was used to explore the effects of gene-environment and gene-gene interaction on intelligence.Results:In the high fluoride group, children''s IQ scores decreased by 2.502 when the UF_Cr level increased by 1.0 mg/L (β = −2.502, 95% confidence interval [CI]:−4.411, −0.593), and the possibility for having “excellent” intelligence decreased by 46.3% (odds ratio = 0.537, 95% CI: 0.290, 0.994). Children with the GG genotype showed increased IQ scores than those with the AA genotype of rs11627387 locus in the high fluoride group (P < 0.05). Interactions between fluoride exposure and MTHFD1 polymorphisms on intelligence were observed (Pinteraction < 0.05).Conclusion:Our findings suggest that excessive fluoride exposure may have adverse effects on children''s intelligence, and changes in children''s intelligence may be associated with the interaction between fluoride and MTHFD1 polymorphisms.     

Clinical and Genetic Characteristics of Korean Patients Diagnosed with Chronic Enteropathy Associated with SLCO2A1 Gene: A KASID Multicenter Study

Background/AimsChronic enteropathy associated with SLCO2A1 gene (CEAS), an inherited disease characterized by nonspecific intestinal ulcers, has emerged in the Japanese population via loss-of-function mutations in the SLCO2A1 gene. We aimed to investigate the clinical and genetic characteristics of Korean patients diagnosed with CEAS.MethodsFrom July 2018 to July 2021, we performed Sanger sequencing of the SLCO2A1 gene in 46 patients with chronic intestinal ulcers. CEAS was confirmed based on known SLCO2A1 mutations. We summarized the clinical characteristics of patients with confirmed CEAS.ResultsFourteen out of 46 patients (30.4%) had genetically confirmed CEAS, and two SLCO2A1 variants were detected (splicing site variant c.940+1G>A and nonsense mutation [p.R603X] in SLCO2A1). Twelve patients (85.7%) were females and the median age at diagnosis of CEAS was 44.5 years. All patients presented with abdominal pain, and 13 patients (92.9%) presented with anemia (median hemoglobin, 9.6 g/dL). Ten patients (71.4%) had hypoalbuminemia (median, 2.7 g/dL). The most commonly involved site was the ileum (13/14, 92.9%). Manifestations of primary hypertrophic osteoarthropathy (PHO), such as digital clubbing, pachydermia, and periostosis were observed in five patients (28.6%) and two male patients and one female patient satisfied all major PHO diagnostic criteria.ConclusionsThe clinical and genetic characteristics of Korean patients with confirmed CEAS were similar to those reported in the literature. CEAS should be considered in the differential diagnosis for patients with unexplained chronic nonspecific ulcers of the small intestine.     

Establishment of a novel prognostic prediction model through bioinformatics analysis for prostate cancer based on ferroptosis-related genes and its application in immune cell infiltration

BackgroundFerroptosis-related genes (FRGs) play vital roles in survival and prognosis of prostate cancer (PCa) patients. We establish a ferroptosis-related prediction model through bioinformatics analysis for overall survival (OS) and disease-free survival (DFS), so as to evaluate the clinical survival status through the characteristics of immune cell infiltration (ICI), which could provide information for treatment monitoring.MethodsAt first, 268 FRGs were obtained from previous studies. Differentially expressed FRGs were identified based on The Cancer Genome Atlas (TCGA) database, and FRG enrichment analysis was performed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). We then performed univariate, least absolute shrinkage and selection operator (LASSO), and multivariate Cox regression analyses to establish OS- and DFS-related prognostic prediction models. The association of the model and clinicopathological features was further analyzed. Subsequently, unique genomic signatures of immune cell subsets were obtained through the KEGG database. Based on specific genes associated with ferroptosis and their association with ICI, immune infiltration was assessed in patients in different risk groups.ResultsWe constructed an OS- and an DFS-prognostic model through bioinformatics analysis. The predicted values of OS and DFS-related models were higher in T3–4 than in T1–2 (P=0.0057, P<0.001), and the predicted value of the DFS model in N0 stage was higher than that in N1 stage (P=0.0136). Results of Single-sample gene set enrichment analysis (ssGSEA) on the basis of the KEGG dataset showed p53 signaling being the most enriched signal in the high-risk group, while endocytosis was the most enriched signal in the low-risk group. M2 macrophages (P=0.007) and neutrophils (P=0.024) were enriched in the high-risk group, and CD4-activated memory T cells were significantly accumulated in the low-risk group (P=0.017).ConclusionsThe OS- and DFS-related model based on FRGs and ICI create new insights into the disease state assessment of PCa patients., which may aid in the development of individualized and precise treatment in the future.     

TRIM27 is an adverse prognostic biomarker and associated with immune and molecular profiles in right-sided colon cancer

Right-sided colon cancer (RCC), as an independent tumor entity, shows a poor prognosis. It is imperative to detect immune microenvironment-related genes for predicting RCC patient prognosis and study their function in RCC. Tripartite motif-containing 27 (TRIM27) was identified as a risk signature from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) datasets by using weighted gene co-expression network analysis, differentially expressed analysis, and univariate Cox analysis. It predicted a poorer overall survival and increased lymph node metastasis, which were then validated in our 48 clinical samples. Using immunohistochemistry, TRIM27 was found to be highly expressed in both cancer cells and surrounding immunocytes, and its expression in tumor or immune cells both predicted a poorer prognosis. Thereafter, the functional mechanism, immune and molecular characteristics of TRIM27 were investigated using gene set enrichment analysis (GSEA), ESTIMATE, CIBERSORT, and gene set variation analysis (GSVA) at the single-cell, somatic mutation, and RNA-seq level. Patients with highly expressed TRIM27 presented lower CD4⁺ T cell infiltration and activation of the mTORC1/glycolysis pathway. In addition, patients with highly expressed TRIM27 were characterized by hypermetabolism, higher tumor purity, more BRAF mutation, and more chromosomal instability. Collectively, TRIM27 is an important immune-related prognostic biomarker in patients with RCC. It may function via activating the mTORC1/glycolysis pathway and suppressing CD4⁺ T cells. These results indicated that TRIM27 could be a promising therapeutic target in RCC.     

大鼠肝移植自然耐受模型移植术后Foxp3基因表达的变化

目的 探讨大鼠肝移植自然耐受模型移植术后Foxp3基因表达的变化及意义.方法 分别建立大鼠急性排斥(DA→LEW)、自然耐受(LEW→DA)、同基因组(DA→DA)肝移植模型,收集术后7、14、28d外周血、脾脏、肝脏标本,FCM检测标本淋巴细胞中CD4 CD25 Tr (regulatory T cell)比例变化,RT-PCR, Western blot分别检测肝脏内Foxp3基因及其scrufin蛋白的表达.结果 术后7d,各组外周血、脾脏、肝脏中CD4 CD25 Tr细胞比例无明显差异.术后14d自然耐受组脾脏、肝脏内CD4 CD25 Tr细胞比例明显高于同基因组(P<0.05),外周血中差异无统计学意义(P>0.05).术后28d自然耐受组与同基因组间无差异(P>0.05).术后7d,自然耐受组移植物Foxp3 mRNA水平明显高于排斥组和同基因组(P<0.05);术后14d,自然耐受组移植物Foxp3 mRNA水平明显高于同基因组(P<0.05);术后28d,自然耐受组与同基因组移植物Foxp3 mRNA表达无差异(P>0.05).自然耐受组与同基因术后14、28d scrufin蛋白条带明显强于7d,且自然耐受组表达强于同基因组.结论 大鼠肝移植术后1～2周内,移植肝内Foxp3基因表达可诱导机体对移植物产生免疫耐受.     

Oestrogen receptor α gene polymorphisms,insomnia, and cognitive functions in perimenopausal and postmenopausal women in non-manual employment

IntroductionA potential way to explain the relationships between sleep disorders and cognitive disorders during menopausal transition is the identification of genetic markers related to changes in cognitive functions, as well as changes in quality of sleep during menopause. The objective was an analysis of the relationship between sleep disorders and cognitive disorders, according to the possessed oestrogen receptor α gene polymorphism (ESR1) in perimenopausal and postmenopausal women.Material and methodsThe study included 300 women aged 44–66 years, employed as non-manual workers. A computerised battery of the Central Nervous System Vital Signs (CNS VS) test was used to diagnose cognitive functions. ESR1 polymorphisms were genotyped using PCR-RFLP methods. The Athens Insomnia Scale was used to diagnose sleep disorders.ResultsMore severe insomnia was related to worse complex memory, visual memory, and simple attention in the total group of examined women. More severe insomnia was related to worse simple attention in women with genotypes AG Xba I or TC Pvu II ESR1, in perimenopausal women with genotypes AG Xba I or TC Pvu II ESR1. During the postmenopausal period, the severity of insomnia negatively correlated with visual memory in carriers of Pvu II TT, and with reaction time in carriers of Xba I AA.ConclusionsThe results indicate an important role of oestrogen receptor α gene polymorphism in the modulation of the effect of insomnia on cognitive functions in peri- and postmenopausal women.     

植物微小RNA跨界调控机制及其应用研究进展

微小RNA (microRNA, miRNA)是在多种真核细胞及病毒中发现的一类参与基因表达调控的非编码单链RNA。近期研究表明,植物来源的miRNA可以稳定地存在于动物的血液与组织器官中,并参与调控多种靶蛋白的表达而发挥作用。本文拟从植物miRNA跨界调控的研究现状出发,梳理植物miRNA发挥跨界调控功能的机制,并对其在中药miRNA活性成分的挖掘、小核酸药物开发、以植物为载体的药物开发等应用前景进行综述,有利于加深对植物miRNA跨界调控的认识以及对药用植物作用机制和生物学功能的理解,为开发预防或治疗人类疾病的新方法提供思路。     

抗PD-1单抗生物学活性测定方法的联合验证

针对抗PD-1单抗的报告基因活性测定法开展实验室间的联合验证,以研究该方法多实验室间的可应用和可转移性。本文采用两种协作方案,分别对该方法的检测内、检测间和实验室间精密度以及线性和准确性进行了研究。结果显示:该方法的检测内精密度的95%可置信区间为(1.72～16.89)%,检测间精密度为(2.63～17.67)%,实验室间精密度为(9.00～14.26)%;线性的相关系数均大于0.99,对于不同效价水平准确性的95%可置信区间, 50%为(91.83～104.40)%, 75%为(90.40～101.40)%, 100%为(94.71～105.60)%, 125%为(94.00～102.00)%, 150%为(96.73～104.30)%。联合验证结果证明,该针对抗PD-1单抗的报告基因测活法,其精密度、线性和准确性均良好,可应用于不同实验室抗PD-1单抗的放行检测及稳定性分析。