Search

Genomic integrity is critical for sexual reproduction, ensuring correct transmission of parental genetic information to the descendant. To preserve genomic integrity, germ cells have evolved multiple DNA repair mechanisms, together termed as DNA damage response. The RNA N6-methyladenosine is the most abundant mRNA modification in eukaryotic cells, which plays important roles in DNA damage response, and YTH N6-methyladenosine RNA binding protein 2 (YTHDF2) is a well-acknowledged N6-methyladenosine reader protein regulating the mRNA decay and stress response. Despite this, the correlation between YTHDF2 and DNA damage response in germ cells, if any, remains enigmatic. Here, by employing a Ythdf2-conditional knockout mouse model as well as a Ythdf2-null GC-1 mouse spermatogonial cell line, we explored the role and the underlying mechanism for YTHDF2 in spermatogonial DNA damage response. We identified that, despite no evident testicular morphological abnormalities under the normal circumstance, conditional mutation of Ythdf2in adult male mice sensitized germ cells, including spermatogonia, to etoposide-induced DNA damage. Consistently, Ythdf2-KO GC-1 cells displayed increased sensitivity and apoptosis in response to DNA damage, accompanied by the decreased SET domain bifurcated 1 (SETDB1, a histone methyltransferase) and H3K9me3 levels. The Setdb1knockdown in GC-1 cells generated a similar phenotype, but its overexpression in Ythdf2-null GC-1 cells alleviated the sensitivity and apoptosis in response to DNA damage. Taken together, these results demonstrate that the N6-methyladenosine reader YTHDF2 promotes DNA damage repair by positively regulating the histone methyltransferase SETDB1 in spermatogonia, which provides novel insights into the mechanisms underlying spermatogonial genome integrity maintenance and therefore contributes to safe reproduction.Loss or reduction of the m6A reader YTHDF2 sensitizes spermatogonia to DNA damage, which is mediated, at least in part, by the H3K9 methyltransferase SETDB1.Graphical Abstract

[Display omitted]

Intensive selective breeding for high growth rate and body weight cause excess abdominal fat in broilers. Gut microbiota and folic acid were reported to regulate lipid metabolism. A total of 210 one-day-old broilers were divided into the control (folic acid at 1.3 mg/kg) and folic acid groups (folic acid at 13 mg/kg) to illustrate the effects of folic acid on growth performance, abdominal fat deposition, and gut microbiota, and the experiment lasted 28 d. Results revealed that dietary folic acid addition decreased abdominal fat percentage (P < 0.05) and down-regulated genes expression related to cell proliferation and differentiation in abdominal fat including IGF1, EGF, C/EBPα, PPARγ, PLIN1, FABP4and PCNA(P < 0.05). Folic acid addition decreased caecal Firmicutes-to-Bacteroidetes ratio (P < 0.01) and increased the proportions of Alistipes, Oscillospira, Ruminococcus, Clostridium, Dehalobacteriumand Parabacteroides(P < 0.05). Caecal acetic acid, and propionic acid contents were found to be higher under folic acid treatment (P < 0.05), which were negatively related to genes expression associated with adipocyte proliferation and differentiation (P < 0.05). Ruminococcuswas positively correlated with caecal acetic acid content, and the same phenomenon was detected between propionic acid and Oscillospiraand Ruminococcus(P < 0.05). Acetic acid and Oscillospirawere identified to be negatively associated with abdominal fat percentage (P < 0.05). In conclusion, our data demonstrated that dietary supplementation of folic acid reduced fat deposition in broilers by inhibiting abdominal adipocyte proliferation and differentiation, which might be mediated by changes in gut microbiota and short chain fatty acid production.

Cytochrome P450 monooxygenases (P450s) play important roles in the detoxification metabolism of xenobiotics and are involved in the resistance of insects to many insecticides. In this study, piperonyl butoxide (PBO), an inhibitor of P450 enzyme activity, significantly increased the toxicity of clothianidin in the clothianidin-resistant (CL-R) population of Bradysia odoriphaga. The enzyme activity of P450 in the CL-R population was significantly higher than that in the SS population. Furthermore, four P450 genes were found to be significantly overexpressed in the CL-R population. Tissue-specific expression analysis indicates that CYP9J57, CYP3828A1, CYP6SX1, and CYP6QE1were most highly expressed in the midgut and/or Malpighian tubules. After exposure to LC30of clothianidin, the expression levels of the four P450 genes were significantly upregulated. The RNAi-mediated knockdown of CYP9J57, CYP3828A1, and CYP6QE1significantly increased the susceptibility of B. odoriphagato clothianidin. These results suggest that P450 genes are involved in clothianidin resistance in B. odoriphaga. This provides a better understanding of P450-mediated clothianidin resistance in B. odoriphagaand will contribute to the management of insect resistance to insecticides.

MicroRNA (miRNA) detection is a critical aspect of disease diagnosis, and recent studies indicate that miRNA-622 could be a potential target for lung cancer. Herein, Cu single atoms were anchored on graphitic carbon nitride (Cu SAs@CN) as a coreaction accelerator applied in luminol-H2O2system, thereby establishing an efficient and sensitive electrochemiluminescence (ECL) biosensor for miRNA-622 detection. Cu SAs@CN was explored to possess excellent enzyme-like activities that promote the generation of abundant reactive oxygen species, which amplified ECL emission. Meanwhile, in order to improve the accuracy and sensitivity for miRNA-622 detection, the highly specific trans-cleavage ability of CRISPR/Cas12a was combined with a catalytic hairpin assembly strategy. Therefore, an ECL biosensor for miRNA-622 detection was systematically constructed as a proof of concept, achieving an ultralow limit of detection of 1.09 fM, and the feasibility was demonstrated in human serum samples. The findings of this research provide a promising strategy to enhance the ECL response using versatile single-atom catalysts, thus advancing the development of ECL biosensing applications.

Table eggs with color-uniformity shell are visually attractive for consumers. Lueyang black-boned chicken (LBC) lays colorful eggs, which is undesirable for sale of table eggs, but provides a segregating population for mapping functional variants affecting eggshell color. SLCO1B3was identified as the causative gene for blue eggs in the Dongxiang and Araucana chickens. The aim of this study is to map functional variants associated with chicken eggshell color in the SLCO1B3. Eggshell color of LBC (n = 383) was measured using the L*a*b color space. SLCO1B3was resequencing using a subset (n = 30) of 383 samples. Linkage disequilibrium among 139 SNP was analyzed. Association of 16 SNP in the SLCO1B3and 8 in CPOX, ALAS1, and ABCG2genes with L*a*b were tested by a polygenic model (LMM) and a polygenic/oligogenic mixed model (BSLMM). Chromatin state annotations were retrieved from the UCSC database. Effect of SLCO1B3variants distributed in mapping and upstream 1.6-kb regions on promoter activities were analyzed using dual-luciferase reporter assay. One hundred and thirty-nine variants maintained low linkage disequilibrium with 80% of r2less than 0.226. Fifteen SLCO1B3variants were significantly associated with a*, of which 1B3_SNP108 was showed the strongest association and the largest effect on a*. In the BSLMM, 1B3_SNP108 alone appeared in the Markov chain Monte Carlo as major variants in 100% of posterior inclusion probability. None of variants in CPOX, ALAS1, and ABCG2were significantly associated with color indexes except that 2 ALAS1variants were associated with L*. 1B3_SNP108 distributes in the Intron4 where 6 active enhancers and 1 ATAC island were enriched. However, 1B3_SNP108-containing constructs showed negligible activities in the reporter assay. No significant differences of activities between haplotypes were found for five 5′-deleted promoter constructs. The data recognizes 1B3_SNP108 as a valuable marker for breeding of eggshell color. Functional variants are localized in the region adjacent to the 1B3_SNP108 due to low linkage disequilibrium in the LBC. Our findings extend the role of SLCO1B3from a causative gene for blue eggs to a major regulator driving continuous variation of LBC eggshell color.

Di-(2-ethylhexyl) phthalate (DEHP) is a widely used commercial plasticizer. DEHP exposure has a negative impact on brain development and cognition, but the mechanisms responsible for DEHP-induced neurotoxicity are not well understood. Here we showed that DEHP exposure increased maleic dialdehyde and reactive oxygen species contents and decreased endogenous superoxide dismutase activity in a mouse neuroblastoma cell line (N2a cell line). DEHP exposure not only induced reduction of neurite outgrowth, but also led to microtubule-associated protein tau hyperphosphorylation and dissociation from microtubules. Furthermore, DEHP exposure decreased the levels of synapsin-1 and postsynaptic density protein 95 (PSD95), which play critical roles in synaptic function. Antioxidant vitamin E pretreatment prevented DEHP-induced abnormalities in the cells. These results indicate that DEHP exposure could induce abnormal action of proteins including tau, synapsin-1 and PSD95, which play critical roles in the synaptic structure and function, and that these alterations might be mediated by peroxidative damage.

The Wiener-type invariants of a connected graph Gare defined as Wf=∑u,v∈V(G)f(dG(u,v)), where f(x) is a nonnegative function on the distance dG(u,v).For integer k≥2,a graph Gis called k-leaf-connected if |V(G)|≥k+1and given any subset S⊆V(G)with |S|=k,Galways has a spanning tree Tsuch that Sis precisely the set of leaves of T. Thus, a graph is 2-leaf-connected if and only if it is Hamilton-connected. In this paper, we present best possible Wiener-type invariants conditions to guarantee a graph to be k-leaf-connected, which extend the corresponding results on Hamilton-connected graphs. As applications, sufficient conditions for a graph to be k-leaf-connected in terms of the distance (distance signless Laplacian, Harary) spectral radius of Gor its complement are also obtained.

Spermatogenesis is the process by which diploid male germ cells propagate and differentiate into haploid flagellated spermatozoa. This highly complicated process is dependent on testicular somatic cells maturation. While the role of these somatic cells in spermatogenesis is relatively well established, knowledge about their development and maturation, particularly at the molecular level is limited. In this study, we profiled the testicular single-cell transcriptomes of Guanzhong black pigs at the age of 7, 30, 60, 90, and 150 days. Five types of Sertoli cells, five types of Leydig cells, and four types of peritubular myoid cells were identified. Histological analysis revealed the changes in proliferation levels and marker gene expressions, and prion-like protein gene (PRND) was identified as a novel marker for Sertoli cells. Additionally, integrated analyses of porcine and human datasets revealed similarities between human and pig testicular somatic cells. Overall, the data obtained in this study contribute to the understanding of testicular development in pigs as a model species.

Malignant melanoma is the most aggressive form of skin cancer. Although various antimelanoma approaches have been used in the clinics to treat the disease over the last three decades, none of the drugs significantly prolonged the survival of metastatic melanoma patients; hence, effective drugs against metastatic melanoma are highly desired. In this study, we explored an antimetastatic melanoma agent derived from traditional Chinese medicinal herbs and found that jatrorrhizine hydrochloride (JH), an active component of the traditional Chinese medicinal herb Coptis chinensis, inhibited the proliferation and neovascularization of C8161 human metastatic melanoma cells. JH suppressed C8161 cell proliferation in a dose-dependent manner, with a half-maximal inhibitory concentration of 47.4±1.6 moll; however, it did not induce significant cellular apoptosis at doses up to 320 moll. Mechanistic studies showed that JH-induced C8161 cell cycle arrest at the G0G1 transition, which was accompanied by overexpression of the cell cycle-suppressive genes p21and p27at higher doses. Moreover, JH reduced C8161 cell-mediated neovascularization in vitroand in vivoand impeded the expression of the gene for VE-cadherin, a key protein in tumor vasculogenic mimicry and angiogenesis. Taken together, the effective inhibitory effects of JH on metastatic melanoma cell proliferation and neovascularization with low toxicity suggest that JH is a potential new antimelanoma drug candidate.

Background It is well documented that trans-membrane pressure drop (TMPD) of hollow-fiber membrane oxygenators (HFMO) may lead to hemolysis, damage to platelets, and systemic inflammatory response. The purpose of this study was to evaluate five commercially available adult oxygenators in terms of pressure drop during normothermic and hypothermic cardiopulmonary bypass (CPB).Materials and Methods In a clinical setting, 5 different adult HFMOs were tested in terms of TMPDs. Forty patients scheduled for cardiac surgery were enrolled in the study and were divided into 5 groups according to the type of oxygenator used during CPB: group J (Maquet Quardox n=8), group A (Affinity NT n=8), group P (Polystan Safe Maxi n=8), group T (Terumo Capiox SX 18 n=8) and group C (COBE APEX-TM n=8). Clinical parameters were collected during CPB, including mean arterial pressure (MAP), pump flow, temperature, hematocrit, duration of CPB, cross-clamp time and bypass time. TMPDs of pre-oxygenator and post-oxygenator pressures were recorded at the start of systemic cooling (T1), 10 min after stable hypothermia at 30°C (T2), and at termination of rewarming before the end of CPB (T3).Results There were no significant differences among the 5 groups in pump-flow rate, temperature, hematocrit, and mean arterial pressure during CPB. TMPDs of group J were the lowest at different time-points (T1, 30.71 ± 8.42mmHg; T2, 25.71 ± 5.41 mmHg; T3, 27.42 ± 13.01 mmHg). Compared to the other 3 groups (P, C and T groups), TMPDs of groups J and A at various time-points were lower (J group compared with the other three groups (p<0.001). Although TMPDs in groups A, P and T during hypothermia were higher than during normothermia and post-rewarming, there was no significant statistical difference (p> 0.05).Conclusions These results suggest that the HFMOs in groups J and A produced significantly lower TMPDs and pre- and post-oxygenator extracorporeal circuit pressures during normothermic and hypothermic CPB.

No relevant conflicts of interest to declare.

Abstract 3267