Induced neural stem cells regulate microglial activation through Akt-mediated upregulation of CXCR4 and Crry in a mouse model of closed head injury.

ABSTRACT: Microglial activation that occurs rapidly after closed head injury may play important and complex roles in neuroinflammation-associated neuronal damage and repair. We previously reported that induced neural stem cells can modulate the behavior of activated microglia via CXCL12/CXCR4 signaling, influencing their activation such that they can promote neurological recovery. However, the mechanism of CXCR4 upregulation in induced neural stem cells remains unclear. In this study, we found that nuclear factor-κB activation induced by closed head injury mouse serum in microglia promoted CXCL12 and tumor necrosis factor-α expression but suppressed insulin-like growth factor-1 expression. However, recombinant complement receptor 2-conjugated Crry (CR2-Crry) reduced the effects of closed head injury mouse serum-induced nuclear factor-κB activation in microglia and the levels of activated microglia, CXCL12, and tumor necrosis factor-α. Additionally, we observed that, in response to stimulation (including stimulation by CXCL12 secreted by activated microglia), CXCR4 and Crry levels can be upregulated in induced neural stem cells via the interplay among CXCL12/CXCR4, Crry, and Akt signaling to modulate microglial activation. In agreement with these in vitro experimental results, we found that Akt activation enhanced the immunoregulatory effects of induced neural stem cell grafts on microglial activation, leading to the promotion of neurological recovery via insulin-like growth factor-1 secretion and the neuroprotective effects of induced neural stem cell grafts through CXCR4 and Crry upregulation in the injured cortices of closed head injury mice. Notably, these beneficial effects of Akt activation in induced neural stem cells were positively correlated with the therapeutic effects of induced neural stem cells on neuronal injury, cerebral edema, and neurological disorders post-closed head injury. In conclusion, our findings reveal that Akt activation may enhance the immunoregulatory effects of induced neural stem cells on microglial activation via upregulation of CXCR4 and Crry, thereby promoting induced neural stem cell-mediated improvement of neuronal injury, cerebral edema, and neurological disorders following closed head injury.

Impact of green space exposure on blood pressure in Guangzhou, China: Mediation by air pollution, mental health, physical activity, and weight status.

Green space exposure has been inversely associated with blood pressure (BP) levels and hypertension risk. However, empirical evidence on the underlying mechanisms are lacking. This study examined the association of green space exposure with BP and hypertension, and assessed the mediating effects by air pollution, mental health, physical activity, and weight status. Survey data from 719 adults, who lived in Guangzhou (China) in 2016, were used. Three area-level green space indicators, including network distance to the nearest park, percentage of green space and Normalized Difference Vegetation Index within a 1 km Euclidean buffer around residence and workplace, were calculated and linked to individual-level BP measurements. Structural equation models were applied to estimate the direct and indirect associations of the various green space indicators on systolic BP (SBP), diastolic BP (DBP), and hypertension, respectively. After adjusting for multiple covariates, longer network distance to green space was directly associated with higher SBP. Compared to the reference group (0-500m), the differences were 0.11 mmHg (95% CI = 0.03 to 0.19, P = 0.006) for 500-1000m, 0.03 mmHg (95% CI = -0.05 to 0.12, P = 0.45) for 1000-1500m, and 0.16 mmHg (95% CI = 0.09 to 0.23, P < 0.001) for >1500m, respectively. The overall and direct associations were significant for all three indicators (distance or density) with or without considering workplace exposure. The association between network distance to green and SBP was partially (18.4%, 95% CI = 0-42.1%) mediated by mental health. There was no statistical evidence that air pollution, physical activity, or weight status mediate the association. Secondary analyses for other indicators and other outcomes showed similar results. Both distance to green space and more green space around residence and workplace were associated with lower BP and lower risk of hypertension in adults living in a Chinese metropolitan. Mental health partly mediated the association.

Linc-NSC affects cell differentiation, apoptosis and proliferation in mouse neural stem cells and embryonic stem cells in vitro and in vivo.

BACKGROUND: Stem cell therapy is a promising therapeutic strategy. In a previous study, we evaluated tumorigenicity by the stereotactic transplantation of neural stem cells (NSCs) and embryonic stem cells (ESCs) from experimental mice. Twenty-eight days later, there was no evidence of tumor formation or long-term engraftment in the NSCs transplantation group. In contrast, the transplantation of ESCs caused tumor formation; this was due to their high proliferative capacity. Based on transcriptome sequencing, we found that a long intergenic non-coding RNA (named linc-NSC) with unknown structure and function was expressed at 1100-fold higher levels in NSCs than in ESCs. This finding suggested that linc-NSC is negatively correlated with stem cell pluripotency and tumor development, but positively correlated with neurogenesis. In the present study, we investigated the specific role of linc-NSC in NSCs/ESCs in tumor formation and neurogenesis. METHODS: Whole transcriptome profiling by RNA sequencing and bioinformatics was used to predict lncRNAs that are widely associated with enhanced tumorigenicity. The expression of linc-NSC was assessed by quantitative real-time PCR. We also performed a number of in vitro methods, including cell proliferation assays, differentiation assays, immunofluorescence assays, flow cytometry, along with in vivo survival and immunofluorescence assays to investigate the impacts of linc-NSC on tumor formation and neurogenesis in NSCs and ESCs. RESULTS: Following the knockdown of linc-NSC in NSCs, NSCs cultured in vitro and those transplanted into the cortex of mice showed stronger survival ability (P < 0.0001), enhanced proliferation(P < 0.001), and reduced apoptosis (P < 0.05); the opposite results were observed when linc-NSC was overexpressed in ESCs. Furthermore, the overexpression of linc-NSC in ECSs induced enhanced apoptosis (P < 0.001) and differentiation (P < 0.01), inhibited tumorigenesis (P < 0.05) in vivo, and led to a reduction in tumor weight (P < 0.0001). CONCLUSIONS: Our analyses demonstrated that linc-NSC, a promising gene-edited target, may promote the differentiation of mouse NSCs and inhibit tumorigenesis in mouse ESCs. The knockdown of linc-NSC inhibited the apoptosis in NSCs both in vitro and in vivo, and prevented tumor formation, revealing a new dimension into the effect of lncRNA on low survival NSCs and providing a prospective gene manipulation target prior to transplantation. In parallel, the overexpression of linc-NSC induced apoptosis in ESCs both in vitro and in vivo and attenuated the tumorigenicity of ESCs in vivo, but did not completely prevent tumor formation.

A novel sub-regional radiomics model to predict immunotherapy response in non-small cell lung carcinoma.

BACKGROUND: Identifying precise biomarkers of immunotherapy response for non-small cell lung carcinoma (NSCLC) before treatment is challenging. This study aimed to construct and investigate the potential performance of a sub-regional radiomics model (SRRM) as a novel tumor biomarker in predicting the response of patients with NSCLC treated with immune checkpoint inhibitors, and test whether its predictive performance is superior to that of conventional radiomics, tumor mutational burden (TMB) score and programmed death ligand-1 (PD-L1) expression. METHODS: We categorized 264 patients from retrospective databases of two centers into training (n = 159) and validation (n = 105) cohorts. Radiomic features were extracted from three sub-regions of the tumor region of interest using the K-means method. We extracted 1,896 features from each sub-region, resulting in 5688 features per sample. The least absolute shrinkage and selection operator regression method was used to select sub-regional radiomic features. The SRRM was constructed and validated using the support vector machine algorithm. We used next-generation sequencing to classify patients from the two cohorts into high TMB (≥ 10 muts/Mb) and low TMB (< 10 muts/Mb) groups; immunohistochemistry was performed to assess PD-L1 expression in formalin-fixed, paraffin-embedded tumor sections, with high expression defined as ≥ 50% of tumor cells being positive. Associations between the SRRM and progression-free survival (PFS) and variant genes were assessed. RESULTS: Eleven sub-regional radiomic features were employed to develop the SRRM. The areas under the receiver operating characteristic curve (AUCs) of the proposed SRRM were 0.90 (95% confidence interval [CI] 0.84-0.96) and 0.86 (95% CI 0.76-0.95) in the training and validation cohorts, respectively. The SRRM (low vs. high; cutoff value = 0.936) was significantly associated with PFS in the training (hazard ratio [HR] = 0.35 [0.24-0.50], P < 0.001) and validation (HR = 0.42 [0.26-0.67], P = 0.001) cohorts. A significant correlation between the SRRM and three variant genes (H3C4, PAX5, and EGFR) was observed. In the validation cohort, the SRRM demonstrated a higher AUC (0.86, P < 0.001) than that for PD-L1 expression (0.66, P = 0.034) and TMB score (0.54, P = 0.552). CONCLUSIONS: The SRRM had better predictive performance and was superior to conventional radiomics, PD-L1 expression, and TMB score. The SRRM effectively stratified the progression-free survival (PFS) risk among patients with NSCLC receiving immunotherapy.

Single-cell sequencing highlights heterogeneity and malignant progression in actinic keratosis and cutaneous squamous cell carcinoma.

Cutaneous squamous cell carcinoma (cSCC) is the second most frequent of the keratinocyte-derived malignancies with actinic keratosis (AK) as a precancerous lesion. To comprehensively delineate the underlying mechanisms for the whole progression from normal skin to AK to invasive cSCC, we performed single-cell RNA sequencing (scRNA-seq) to acquire the transcriptomes of 138,982 cells from 13 samples of six patients including AK, squamous cell carcinoma in situ (SCCIS), cSCC, and their matched normal tissues, covering comprehensive clinical courses of cSCC. We identified diverse cell types, including important subtypes with different gene expression profiles and functions in major keratinocytes. In SCCIS, we discovered the malignant subtypes of basal cells with differential proliferative and migration potential. Differentially expressed genes (DEGs) analysis screened out multiple key driver genes including transcription factors along AK to cSCC progression. Immunohistochemistry (IHC)/immunofluorescence (IF) experiments and single-cell ATAC sequencing (scATAC-seq) data verified the expression changes of these genes. The functional experiments confirmed the important roles of these genes in regulating cell proliferation, apoptosis, migration, and invasion in cSCC tumor. Furthermore, we comprehensively described the tumor microenvironment (TME) landscape and potential keratinocyte-TME crosstalk in cSCC providing theoretical basis for immunotherapy. Together, our findings provide a valuable resource for deciphering the progression from AK to cSCC and identifying potential targets for anticancer treatment of cSCC.

Alpha-synuclein null mutation exacerbates the phenotype of a model of Menkes disease in female mice.

Genetic modifier screens provide a useful tool, in diverse organisms from Drosophila to C. elegans and mice, for recovering new genes of interest that may reduce or enhance a phenotype of interest. This study reports a modifier screen, based on N-ethyl-N-nitrosourea (ENU) mutagenesis and outcrossing, designed to increase understanding of the normal function of murine α-synuclein (Snca). Human SNCA was the first gene linked to familial Parkinson's disease. Since the discovery of the genetic link of SNCA to Parkinson's nearly three decades ago, numerous studies have investigated the normal function of SNCA protein with divergent roles associated with different cellular compartments. Understanding of the normal function of murine Snca is complicated by the fact that mice with homozygous null mutations live a normal lifespan and have only subtle synaptic deficits. Here, we report that the first genetic modifier (a sensitized mutation) that was identified in our screen was the X-linked gene, ATPase copper transporting alpha (Atp7a). In humans, mutations in Atp7a are linked to to Menkes disease, a disease with pleiotropic phenotypes that include a severe neurological component. Atp7a encodes a trans-Golgi copper transporter that supplies the copper co-factor to enzymes that pass through the ER-Golgi network. Male mice that carry a mutation in Atp7a die within 3 weeks of age regardless of Snca genotype. In contrast, here we show that Snca disruption modifies the phenotype of Atp7a in female mice. Female mice that carry the Atp7a mutation, on an Snca null background, die earlier (prior to 35 days) at a significantly higher rate than those that carry the Atp7a mutation on a wildtype Snca background ATPase copper transporting alpha. Thus, Snca null mutations sensitize female mice to mutations in Atp7a, suggesting that Snca protein may have a protective effect in females, perhaps in neurons, given the co-expression patterns. Although data has suggested diverse functions for human and mouse α-synuclein proteins in multiple cell compartments, this is the first demonstration via use of genetic screening to demonstrate that Snca protein may function in the ER-Golgi system in the mammalian brain in a sex-dependent manner.

Revealing the chemical differences and their application in the storage year prediction of Qingzhuan tea by SWATH-MS based metabolomics analysis.

It's generally believed that the longer the storage, the better the quality of dark tea, but the chemical differences of Qingzhuan tea (QZT) with different storage years is still unclear. Herein, in this work, an untargeted metabolomic approach based on SWATH-MS was established to investigate the differential compounds of QZT with 0-9 years' storage time. These QZT samples were roughly divided into two categories by principal component analysis (PCA). After orthogonal projections to latent structures discriminant analysis (OPLS-DA), 18 differential compounds were putatively identified as chemical markers for the storage year variation of QZT. Heatmap visualization showed that the contents of catechins, fatty acids, and some phenolic acids significantly reduced, flavonoid glycosides, triterpenoids, and 8-C N-ethyl-2-pyrrolidinone-substituted flavan-3-ols (EPSFs) increased with the increase of storage time. Furthermore, these chemical markers were verified by the peak areas corresponding to MS2 ions from SWATH-MS. Based on the extraction chromatographic peak areas of MS and MS2 ions, a duration time prediction model was built for QZT with correlation coefficient R2 of 0.9080 and 0.9701, and RMSEP value of 0.85 and 1.24, respectively. This study reveals the chemical differences of QZT with different storage years and provides a theoretical basis for the quality evaluation of stored dark tea.

Prognostic role of IL-8 in cancer patients treated with immune checkpoint inhibitors: a system review and meta-analysis.

Background: Immune checkpoint inhibitors (ICIs) have been proven to be an effective treatment strategy for a variety of malignant tumors. However, only a subset of patients can benefit from ICIs due to factors such as drug resistance. Therefore, it is crucial to identify biomarkers that can accurately predict the efficacy of ICIs and provide a basis for individualized immunotherapy. In this study, we conducted a systematic review and meta-analysis to explore whether the chemokine interleukin 8 (IL-8) can be used as a biomarker to evaluate the efficacy of ICIs treatment. Methods: We conducted a comprehensive search of several databases, including PubMed, Embase, Web of Science, and Cochrane, to identify relevant articles published up to June 08, 2023. Our inclusion criteria were limited to cohort studies and clinical trials that reported hazard ratios (HR) and 95% confidence intervals (CI) for overall survival (OS) and/or progression-free survival (PFS), as well as the objective response rate (ORR), in cancer patients with high and low IL-8 expression. For data analysis, we used Revman to generate forest plots, subgroup analysis, and assess publication bias. Additionally, Stata was utilized for sensitivity analysis and further examination of publication bias. Results: A total of 24 datasets, involving 3190 participants, were selected from 14 studies. The meta-analysis revealed a reduction in ORR, OS, and/or PFS in the high IL-8 group after treatment with ICIs compared to the low IL-8 group. Conclusion: IL-8 can serve as a biomarker for predicting the efficacy of ICIs. Patients with lower expression of IL-8 may benefit from ICIs treatment. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=383188, identifier CRD42022383188.

Allicin-Loaded Intelligent Hydrogel Coating Improving Vascular Implant Performance.

Coronary atherosclerosis is closely related to inflammation and oxidative stress. Owing to poor biocompatibility, lack of personalized treatment, and late toxic side effects, traditional drug-eluting stent intervention, releasing antiproliferative drugs, can delay endothelial repair and cause late thrombosis. The inflammation caused by atherosclerosis results in an acidic microenvironment and oxidative stress, which can be considered as triggers for precise and intelligent treatment. Here, we used catechol hyaluronic acid (C-HA) and cystamine (Cys) to prepare C-HA-Cys hydrogel coatings by amide reaction. The H2S-releasing donor allicin was loaded in the hydrogel to form an intelligent biomimetic coating. The disulfide bond of Cys made the cross-linked network redox-responsive to the inflammation and oxidative stress in the microenvironment by releasing the drug and H2S intelligently to combat the side effects of stent implantation. This study evaluated the hemocompatibility, anti-inflammatory capacity, vascular wall cytocompatibility, and in vivo histocompatibility of this intelligent hydrogel coating. Furthermore, the effect of H2S released from the coating on atherosclerosis-related signaling pathways such as CD31 and cystathionine γ-lyase (CSE), CD36, and ACAT-1 was investigated. Our results indicate that the C-HA-Cys-Allicin hydrogel coating could be manufactured on the surface of vascular interventional devices to achieve a precise response to the microenvironment of the lesion to release drug, which can attain the purpose of prevention of in-stent restenosis and ensure the effectiveness and safety of the application of interventional devices.

Deubiquitinase OTUD5 as a Novel Protector against 4-HNE-Triggered Ferroptosis in Myocardial Ischemia/Reperfusion Injury.

Despite the development of advanced technologies for interventional coronary reperfusion after myocardial infarction, a substantial number of patients experience high mortality due to myocardial ischemia-reperfusion (MI/R) injury. An in-depth understanding of the mechanisms underlying MI/R injury can provide crucial strategies for mitigating myocardial damage and improving patient survival. Here, it is discovered that the 4-hydroxy-2-nonenal (4-HNE) accumulates during MI/R, accompanied by high rates of myocardial ferroptosis. The loss-of-function of aldehyde dehydrogenase 2 (ALDH2), which dissipates 4-HNE, aggravates myocardial ferroptosis, whereas the activation of ALDH2 mitigates ferroptosis. Mechanistically, 4-HNE targets glutathione peroxidase 4 (GPX4) for K48-linked polyubiquitin-related degradation, which 4-HNE-GPX4 axis commits to myocyte ferroptosis and forms a positive feedback circuit. 4-HNE blocks the interaction between GPX4 and ovarian tumor (OTU) deubiquitinase 5 (OTUD5) by directly carbonylating their cysteine residues at C93 of GPX4 and C247 of OTUD5, identifying OTUD5 as the novel deubiquitinase for GPX4. Consequently, the elevation of OTUD5 deubiquitinates and stabilizes GPX4 to reverse 4-HNE-induced ferroptosis and alleviate MI/R injury. The data unravel the mechanism of 4-HNE in GPX4-dependent ferroptosis and identify OTUD5 as a novel therapeutic target for the treatment of MI/R injury.

Advances in Polysaccharide Production Based on the Co-Culture of Microbes.

Microbial polysaccharides are natural carbohydrates that can confer adhesion capacity to cells and protect them from harsh environments. Due to their various physiological activities, these macromolecules are widely used in food, medicine, environmental, cosmetic, and textile applications. Microbial co-culture is an important strategy that is used to increase the production of microbial polysaccharides or produce new polysaccharides (structural alterations). This is achieved by exploiting the symbiotic/antagonistic/chemo-sensitive interactions between microbes and stimulating the expression of relevant silent genes. In this article, we review the performance of polysaccharides produced using microbial co-culture in terms of yield, antioxidant activity, and antibacterial, antitumor, and anti-inflammatory properties, in addition to the advantages and application prospects of co-culture. Moreover, the potential for microbial polysaccharides to be used in various applications is discussed.

Seizures associated with antibiotics: a real-world disproportionality analysis of FAERS database.

BACKGROUND: Drug-induced seizures are a common occurrence in clinical practice, with research indicating that around 6% of initial seizures are due to drug toxicity. The use of antibiotics is one such cause of drug-related seizures. Previous systematic review has identified specific antibiotics that pose a risk of seizures, but a comprehensive analysis of a large patient sample is needed to determine the risk associated with various drugs. OBJECTIVE: This study aimed to evaluate the association between seizures and various antibiotics that are presently accessible. METHODS: To identify potential risk signals from the US Food and Drug Administration adverse event reporting system (FAERS) database, a disproportionality analysis was conducted. The reporting odds ratio (ROR) using the frequency approach and the information component (IC) using the Bayesian approach were used to detect signals. The median time-to-onset of seizure, as well as the Weibull distribution parameters were calculated to analyze the onset time. RESULTS: A total of 14,407,157 FAERS reports were analyzed.10 antibiotics were associated with seizures that were defined by 41 preferred terms. Onset time were aligned with the wear out failure type profile. CONCLUSION: This study identified 10 antibiotics that showed significant associations with seizures. Imipenem-cilastatin had the highest seizure ROR.

Impact of migration on oral health outcomes of children in multi-beneficial kindergartens in Nanning, Southern China: a cross-sectional study.

OBJECTIVE: To explore the effect of children's migration on their oral health outcomes in multi-beneficial kindergartens in Jiangnan District, Nanning, China, and to provide a basis for improving the oral health of migrant children. METHODS: A cross-sectional study was conducted among 470 children aged 5 years in Jiangnan District, Nanning, Guangxi. A questionnaire was used to collect information on their demographic and socioeconomic background, migration experience, eating habits, oral hygiene behaviours and utilization of dental care services. Dental caries of primary teeth was examined using the decayed, missing, and filled teeth (dmft) index recommended by the World Health Organization. Dental caries experience and oral health-related behaviours were compared between migrant and resident children. The impact of children's migration attributes on their oral health outcomes was examined by univariate and multivariate analyses. RESULTS: Among the examined children, 52.3% were migrant children. The prevalence of caries among the children in multi-beneficial kindergartens was 78.3%, and the mean number of dmft was 5.73 ± 5.00. The prevalence of caries was 81.7% for migrant children and 74.6% for resident children (p = 0.060). No significant difference was found in the mean numbers of DMFT between migrant children and resident children (5.96 ± 4.81 vs. 5.47 ± 5.20, p = 0.139). There were significant differences in the frequency of tooth brushing (p = 0.023) and parental help with tooth brushing (p = 0.008), typical use of fluoride (p = 0.012), regular dental check-ups (p = 0.003) and experience of dental fillings for caries (p < 0.001) between migrant and resident children. The multivariate logistic regression analysis showed that among the children with caries, the proportion of resident children who had regular dental check-ups was 1.720 times higher than that of migrant children (95% CI = 1.155 ~ 2.560), and resident children were more likely to have caries filled than migrant children (OR = 3.313, 95% CI = 1.585 ~ 6.927). CONCLUSION: Oral health status and oral health behaviours were poor among children in multi-beneficial kindergartens in Nanning, China, and migration might be a significant predictive indicator for the poor utilization of dental care services by children. The government departments should make special policy to promote the children's oral health in multi-beneficial kindergartens, and invest more to cover the migrant children's utilization of oral health services.

Progress, challenge and prospect of plant plastome annotation.

The plastome (plastid genome) represents an indispensable molecular data source for studying phylogeny and evolution in plants. Although the plastome size is much smaller than that of nuclear genome, and multiple plastome annotation tools have been specifically developed, accurate annotation of plastomes is still a challenging task. Different plastome annotation tools apply different principles and workflows, and annotation errors frequently occur in published plastomes and those issued in GenBank. It is therefore timely to compare available annotation tools and establish standards for plastome annotation. In this review, we review the basic characteristics of plastomes, trends in the publication of new plastomes, the annotation principles and application of major plastome annotation tools, and common errors in plastome annotation. We propose possible methods to judge pseudogenes and RNA-editing genes, jointly consider sequence similarity, customed algorithms, conserved domain or protein structure. We also propose the necessity of establishing a database of reference plastomes with standardized annotations, and put forward a set of quantitative standards for evaluating plastome annotation quality for the scientific community. In addition, we discuss how to generate standardized GenBank annotation flatfiles for submission and downstream analysis. Finally, we prospect future technologies for plastome annotation integrating plastome annotation approaches with diverse evidences and algorithms of nuclear genome annotation tools. This review will help researchers more efficiently use available tools to achieve high-quality plastome annotation, and promote the process of standardized annotation of the plastome.

Effects of micro(nano)plastics on the reproductive system: A review.

Microplastics (100nm-5 mm) and nanoplastics (1-100 nm) are collectively referred to as micro(nano)plastics (MNPs), which are refractory to degradation, easy to migration, small in size, strong in adsorption, and can widely present in human living environment. A number of studies have confirmed that MNPs can be exposed to the human body through a variety of routes, and can penetrate various barriers to enter the reproductive system, suggesting that MNPs may pose potential harm to human reproductive health. Current studies most were limited to phenotypic studies and their subjects were basically lower marine organisms and mammals. Therefore, in order to provide theoretical base for further exploring the effects of MNPs on the human reproductive system, this paper searched the relevant literature at home and abroad, mainly analyzed rodent experiments, and concluded that the main exposure routes of MNPs are dietary intake, air inhalation, skin contact and medical plastics. After entering the reproductive system, MNPs produce reproductive toxicity mainly through oxidative stress, inflammation, metabolic disorders, cytotoxicity and other mechanisms. More work is required to comprehensively identify the exposure routes, improve the detection methods to evaluate the effective exposure and deeply study the specific mechanisms of toxic effects, withing the aim of conducting relevant studies at the population level in the next step.

A theoretical base for non-invasive prenatal paternity testing.

There is an increasing demand for prenatal paternity testing in the forensic applications, which identify biological fathers before the birth of children. Currently, one of the most effective and safe Non-Invasive Prenatal Paternity Testing (NIPPT) methods is high-throughput Next-Generation Sequencing (NGS)-based SNP genotyping of cell-free DNA in maternal peripheral blood. To the best of our knowledge, nearly all methods being used in such applications are based on traditional postnatal paternity tests and/or statistical models of conventional polymorphism sites. These methods have shown unsatisfactory performance due to the uncertainty of fetal genotype. In this study, we propose a cutting-edge methodology called the Prenatal paternity Test Analysis System (PTAS) for cell-free fetal DNA-based NIPPT using NGS-based SNP genotyping. With the implementation of our proposed PTAS methodology, 63 out of 64 early-pregnancy (i.e., less than seven weeks) samples can be precisely identified to determine paternity, except for one sample that does not meet quality control requirements. Although the fetal fraction of the non-identified sample is extremely low (0.51%), its paternity can still be detected by our proposed PTAS methodology through unique molecular identifier tagging. Paternity of the total 313 samples for mid-to-late pregnancy (i.e., more than seven weeks) can be accurately identified. Extensive experiments indicate that our methodology makes a significant breakthrough in the NIPPT theory and will bring substantial benefits to forensic applications.

KISS1 Gene Variations and Susceptibility to Idiopathic Recurrent Pregnancy Loss.

It is widely accepted that kisspeptin plays an integral role in the regulation of reproduction. Genetic variations in the KISS1 gene have been frequently reported to be linked to reproductive diseases, but there is still a lack of data on the association between KISS1 variations and female reproductive disorders. The present study aimed to examine the association of three missense SNPs in the KISS1 gene including rs12998, rs35431622, and rs4889 in association with idiopathic recurrent pregnancy loss (iRPL). A total of 720 individuals were recruited in this study. The DNA from the blood sample was extracted and genotyped using the PCR method. Haplotype and linkage disequilibrium (LD) have also been analyzed. The results of this study suggested that rs12998 G > A and rs4889 C > G had a significant association with iRPL (p < 0.05); while rs35431622 A > G didn't indicate any association with iRPL. A significant association was also found for three haplotypes including C-A-A, G-G-G, and G-G-A in this population. The analysis also showed a significant LD between rs12998 and rs35431622 (P < 0.0005). The rs12998 G > A and rs4889 C > G variants of KISS1 are linked to unexplained recurrent pregnancy loss and may be risk factors for this disease.