Water-in-water emulsions stabilized by nano-chitin.

A water-in-water (W/W) emulsion consists of microdroplets was formed by the spontaneous liquid-liquid separation by mixing polyacrylic acid and chitosan oligosaccharide in water, and these microdropletes were stabilized by nano-chitin, formed water-in-water Pickering emulsions. By taking the advantage of interfacial adsorption of nano-chitin, the W/W emulsion droplets composed of polyacrylic acid/chitosan oligosaccharide (COS/PAA) polyelectrolyte coacervate were successfully stabilized. Research results indicated that composite microspheres were formed by the nano-chitin stabilized COS/PAA emulsion, and the size of these composite microspheres was related to the concentration and morphology of the nano-chitin. As the concentration of nano-chitin increases, the size of the composite microspheres first increases and then decreases, gradually becoming more uniform; whereas a decrease in the length of nano-chitin will lead to an increase in the size of the composite microspheres. The formation of composite microspheres may be due to the electrostatic interactions between nano-chitin and the emulsion droplets. In addition, the composite microspheres exhibit the best release effect for berberine hydrochloride at a pH value of 3.

Percutaneous or surgical LAAO for stroke prevention in patients with atrial fibrillation: A network meta-analysis.

Background: Stroke, which is mainly caused by thrombus formation in the left atrial appendage, represents the most prevalent complication of atrial fibrillation (AF). Both percutaneous left atrial appendage occlusion (p-LAAO) and surgical LAAO (s-LAAO) are used to treat AF and prevent stroke events. However, no head-to-head randomized controlled trials (RCTs) compared these strategies. Objective: To examine the efficacy and safety of diverse strategies for reducing stroke risk using a network meta-analysis (NMA). Methods: PubMed, EMBASE, and Cochrane repositories were explored to identify RCTs involving p-LAAO or s-LAAO, and five were included for NMA. This investigation adhered to the guidelines outlined in the Preferred Reporting Items for Systematic Reviews and Meta-analyses. The NMA was pooled using the Bayesian random effect framework. All findings were expressed as odds ratios accompanied by a 95 % confidence interval.The primary efficacy endpoint was any stroke (AS), and the secondary efficacy endpoint was combined AS and systematic embolism (AS/SE). The primary and secondary safety endpoints were major bleeding (MB) and all-cause death (ACD), respectively. Results: Our meta-analysis incorporated 6337 individuals diagnosed with AF. The NMA demonstrated a reducing trend in AS and AS/SE for s-LAAO versus p-LAAO, while p-LAAO showed a benefit in reducing MB and ACD. Conclusions: and Relevance: s-LAAO could potentially benefit individuals at elevated risk for stroke, whereas p-LAAO may be linked to a reduced likelihood of bleeding.

Anomaly Detection Method for Industrial Control System Operation Data Based on Time-Frequency Fusion Feature Attention Encoding.

Anomaly detection in industrial control system (ICS) data is one of the key technologies for ensuring the security monitoring of ICSs. ICS data are characterized as complex, multi-dimensional, and long-sequence time-series data that embody ICS business logic. Due to its complex and varying periodic characteristics, as well as the presence of long-distance and misaligned temporal associations among features, current anomaly detection methods in ICS are insufficient for feature extraction. This paper proposes an anomaly detection method named TFANet, based on time-frequency fusion feature attention encoding. Considering that periodic variations are more concentrated in the frequency domain, this method first transforms the time-domain data into the frequency domain, obtaining both amplitude and phase data. Then, these data, together with the original time-series data, are used to extract features from two perspectives: long-term temporal changes and long-distance associations. Finally, the six features learned from both the time and frequency domains are fused, and the feature weights are calculated using an attention mechanism to complete the anomaly classification. In multi-classification tasks on three ICS datasets, the proposed method outperforms three popular time-series models-iTransformer, Crossformer, and TimesNet-across five metrics: accuracy, precision, recall, F1 score, and AUC-ROC, with average improvements of approximately 19%, 37%, 31%, 35%, and 22%, respectively.

A novel macroporous carboxymethyl chitosan/sodium alginate sponge dressing capable of rapid hemostasis and drug delivery.

Carboxymethyl chitosan (CMCS) and sodium alginate (SA), which are excellent polysaccharide-based hemostatic agents, are capable of forming polyelectrolyte complexes (PEC) through electrostatic interactions. However, CMCS/SA PEC sponges prepared by the conventional sol-gel process exhibited slow liquid absorption rate and poor mechanical properties post-swelling. In this work, a novel strategy involving freeze casting followed by acetic acid vapor treatment to induce electrostatic interactions was developed to fabricate novel PEC sponges with varying CMCS/SA mass ratios. Compared to sol-gel process sponge, the novel sponge exhibited a higher density of electrostatic interactions, resulting in denser pore walls that resist re-gelation and swelling according to FTIR, XRD, and SEM analyses. Additionally, the liquid absorption kinetics, as well as compression and tension tests, demonstrated that the novel sponge had significantly improved rapid blood absorption capacity and mechanical properties. Furthermore, in vitro coagulation and drug release studies showed that the novel sponge had a lower blood clotting index and clotting time, along with a slower drug release rate after loading with berberine hydrochloride, showcasing its potential as a rapid hemostatic dressing with controlled drug release capabilities.

Cadmium Exposure in Male Rats Results in Ovarian Granulosa Cell Apoptosis in Female Offspring and Paternal Genetic Effects.

The aim of this study was to investigate whether the damage to male offspring induced by cadmium (Cd) exposure during embryonic period leads to the apoptosis of ovarian granulosa cells (OGCs) in the next generation of female offspring, and whether this apoptosis in the offspring was due to paternal genetic effects. Pregnant Sprague-Dawley (SD) rats were exposed to CdCl2 (0, 0.5, 2.0, or 8.0 mg/kg) by gavage daily for 20 days to produce the filial 1 (F1) generation. F1 males were mated with newly purchased females to produce the F2 generation, and the F3 generation was generated in the same way. No apoptotic bodies were observed in the OGCs of either the F2 or F3 generation as shown by electron microscopy, and a reduced OGC apoptosis rate (detected by flow cytometry) was observed in F2 OGCs from the Cd-exposed group. Moreover, the mRNA (qRT-PCR) levels of Bax and Bcl-2 and the protein (western blotting) level of pro-caspase-8 increased in the F2 generation (p < 0.05). The expression of apoptosis-related miRNAs (qRT-PCR) and methylation of apoptosis-related genes (determined via bisulfite-sequencing PCR) in OGCs were further determined. Compared with those of the controls, the expression patterns of microRNAs (miRNAs) in the F2 offspring were different in the Cd-exposed group. The miR-92a-2-5p expression levels were decreased in both the F2 and F3 generations (p < 0.05), while the average methylation level of apoptosis-related genes did not change significantly (except for individual loci). In summary, this study showed that the paternal genetic intergenerational effect of male Cd exposure during embryonic period induced apoptosis of OGCs in the offspring was weakened, and the transgenerational effect disappeared; nevertheless, intergenerational and transgenerational changes in apoptosis-related genes, epigenetic modifications, DNA methylation, and miRNAs were observed, and may be important for understanding the homeostatic mechanisms of the body to alleviate the intergenerational transmission of Cd-induced damage.

Acoustofluidics-enhanced biosensing with simultaneously high sensitivity and speed.

Simultaneously achieving high sensitivity and detection speed with traditional solid-state biosensors is usually limited since the target molecules must passively diffuse to the sensor surface before they can be detected. Microfluidic techniques have been applied to shorten the diffusion time by continuously moving molecules through the biosensing regions. However, the binding efficiencies of the biomolecules are still limited by the inherent laminar flow inside microscale channels. In this study, focused traveling surface acoustic waves were directed into an acoustic microfluidic chip, which could continuously enrich the target molecules into a constriction zone for immediate detection of the immune reactions, thus significantly improving the detection sensitivity and speed. To demonstrate the enhancement of biosensing, we first developed an acoustic microfluidic chip integrated with a focused interdigital transducer; this transducer had the ability to capture more than 91% of passed microbeads. Subsequently, polystyrene microbeads were pre-captured with human IgG molecules at different concentrations and loaded for detection on the chip. As representative results, ~0.63, 2.62, 11.78, and 19.75 seconds were needed to accumulate significant numbers of microbeads pre-captured with human IgG molecules at concentrations of 100, 10, 1, and 0.1 ng/mL (~0.7 pM), respectively; this process was faster than the other methods at the hour level and more sensitive than the other methods at the nanomolar level. Our results indicated that the proposed method could significantly improve both the sensitivity and speed, revealing the importance of selective enrichment strategies for rapid biosensing of rare molecules.

Precisely Regulating Asymmetric Charge Distribution by Single-Atom Central Doped Ag-Based Series Clusters for Enhanced Photoreduction of CO2 to Alcohol Fuels.

High efficiently photocatalytic CO2 reduction (CO2RR) into liquid fuels in pure water system remains challenged. Iron polyphthalocyanine (FePPc) with strong light harvesting, unique Fe-N4 structure, abundant pores, and good stability could serve as a promising catalyst for CO2 photoreduction. To further improve the catalytic efficiency, herein, symmetry-breaking Fe sites are constructed by coupling with atomically precise M1Ag24 (M=Ag, Au, Pt) series clusters. Especially, the introduction of Pt1Ag24 causes the most asymmetric charge distribution of Fe in FePPc (followed by Au1Ag24 and Ag25), leading to the favorable CO2 adsorption and activation. In addition, Pt1Ag24-FePPc exhibits the most effective photogenerated carriers transfer and separation. As a result, Pt1Ag24-FePPc shows the methanol/ethanol yield of 48.55/32.97 µmol·gcat-1·h-1 in H2O-CO2 system under visible light irradiation, ~ 1.65/1.25-fold, 1.83/1.37-fold, and 3.6/1.61-fold higher than that of Au1Ag24-FePPc, Ag25-FePPc, and FePPc, respectively. This work provides a concept for precisely construction and regulation symmetry-breaking sites of cluster-based catalysts for effective CO2 conversion.

A Manganese-Based Nanodriver Coordinates Tumor Prevention and Suppression through STING Activation in Glioblastoma.

Glioblastoma (GBM), the most prevalent and aggressive primary malignant brain tumor, exhibits profound immunosuppression and demonstrates a low response rate to current immunotherapy strategies. Manganese cations (Mn2+) directly activate the cGAS/STING pathway and induce the unique catalytic synthesis of 2'3'-cGAMP to facilitate type I IFN production, thereby enhancing innate immunity. Here, a telodendrimer and Mn2+-based nanodriver (PLHM) with a small size is developed, which effectively target lymph nodes through the blood circulation and exhibit tumor-preventive effects at low doses of Mn2+ (3.7 mg kg-1). On the other hand, the PLHM nanodriver also exhibits apparent antitumor effects in GBM-bearing mice via inducing in vivo innate immune responses. The combination of PLHM with doxorubicin nanoparticles (PLHM-DOX NPs) results in superior inhibition of tumor growth in GBM-bearing mice due to the synergistic potentiation of STING pathway functionality by Mn2+ and the presence of cytoplasmic DNA. These findings demonstrate that PLHM-DOX NPs effectively stimulate innate immunity, promote dendritic cell maturation, and orchestrate cascaded infiltration of CD8 cytotoxic T lymphocytes within glioblastomas characterized by low immunogenicity. These nanodivers chelated with Mn2+ show promising potential for tumor prevention and antitumor effects on glioblastoma by activating the STING pathway.

The study of double-network carboxymethyl chitosan/sodium alginate based cryogels for rapid hemostasis in noncompressible hemorrhage.

Developing an injectable hemostatic dressing with shape recovery and high blood absorption ratio for rapid hemostasis in noncompressible hemorrhage maintains a critical clinical challenge. Here, double-network cryogels based on carboxymethyl chitosan, sodium alginate, and methacrylated sodium alginate were prepared by covalent crosslinking and physical crosslinking, and named carboxymethyl chitosan/methacrylated sodium alginate (CM) cryogels. Covalent crosslinking was achieved by methacrylated sodium alginate in the freeze casting process, while physical crosslinking was realized by electrostatic interaction between the amino group of carboxymethyl chitosan and the carboxyl group of sodium alginate. CM cryogels exhibited large water swelling ratios (8167 ± 1062 %), fast blood absorption speed (2974 ± 669 % in 15 s), excellent compressive strength (over 160 kPa for CM100) and shape recovery performance. Compared with gauze and commercial gelatin sponge, better hemostatic capacities were demonstrated for CM cryogel with the minimum blood loss of 40.0 ± 8.9 mg and the lowest hemostasis time of 5.0 ± 2.0 s at hemostasis of rat liver. Made of natural polysaccharides with biocompatibility, hemocompatibility, and cytocompatibility, the CM cryogels exhibit shape recovery and high blood absorption rate, making them promising to be used as an injectable hemostatic dressing for rapid hemostasis in noncompressible hemorrhage.

Continuous immunosuppression is required for suppressing immune responses to xenografts in non-human primate brains.

Continuous immunosuppression has been widely used in xenografts into non-human primate brains. However, how immune responses change after transplantation in host brains under continuous immunosuppressive administration and whether immunosuppression can be withdrawn to mitigate side effects remain unclear. Human induced neural stem/progenitor cells (iNPCs) have shown long-term survival and efficient neuronal differentiation in primate brains. Here, we evaluate the immune responses in primate brains triggered by human grafts. The results show that the immune responses, including the evident activation of microglia and the strong infiltration of lymphocytes (both T- and B-cells), are caused by xenografts at 4 months post transplantation (p.t.), but significantly reduced at 8 months p.t. under continuous administration of immunosuppressant Cyclosporin A. However, early immunosuppressant withdrawal at 5 months p.t. results in severe immune responses at 10 months p.t. These results suggest that continuous long-term immunosuppression is required for suppressing immune responses to xenografts in primate brains.

In Vitro Detection of S100B and Severity Evaluation of Traumatic Brain Injury Based on Biomimetic Peptide-Modified Nanochannels.

The diagnosis and evaluation of traumatic brain injury (TBI) are crucial steps toward the treatment and prognosis of patients. A common question remains as to whether it is possible to introduce an ideal device for signal detection and evaluation that can directly connect digital signals with TBI, thereby enabling prompt response of the evaluation signal and sensitive and specific functioning of the detection process. Herein, a method is presented utilizing polymetric porous membranes with TRTK-12 peptide-modified nanochannels for the detection of S100B (a TBI biomarker) and assessment of TBI severity. The method leverages the specific bonding force between TRTK-12 peptide and S100B protein, along with the nanoconfinement effect of nanochannels, to achieve high sensitivity (LOD: 0.002 ng mL-1) and specificity (∆I/I0: 44.7%), utilizing ionic current change as an indicator. The proposed method, which is both sensitive and specific, offers a simple yet responsive approach for real-time evaluation of TBI severity. This innovative technique provides valuable scientific insights into the advancement of future diagnostic and therapeutic integration devices.

Rapid Yet Efficient Reduction of Graphene Oxide Triggered by Semi-Molten Metals.

Reduced graphene oxide (rGO) has garnered extensive attention as electrodes, sensors, and membranes, necessitating the efficient reduction of graphene oxide (GO) for optimal performance. In this work, a swift reduction of GO that involves bringing GO foam in contact with semi-molten metals like tin (Sn) and lithium (Li) is presented. These findings reveal that the electrical resistance of GO foam is significantly diminished by its interaction with these metals, even in dry air. Taking inspiration from this technique, Sn foil is employed to encase the GO foam, followed by a calcination in 15 vol% H2 /Ar environment at 235 °C to fabricate the rGO, which demonstrates a remarkably lower electrical resistivity of 0.42 Ω cm when compared to the chemically reduced GO via hydrazine hydrate (650 Ω cm). The reduction mechanism entails the migration of Sn on GO and its subsequent reaction with oxygen functional groups. SnO/Sn(OH)2 formed from the reaction can be subsequently reversed through reduction by H2 to Sn. Utilizing this rGO as the host material for a sulfur cathode, a lithium-sulfur battery is constructed that displays a specific capacity of 1146 mAh g-1 and maintains a capacity retention of 68.4% after 300 cycles at a rate of 0.2 C.

Effects of cadmium exposure during pregnancy on genome-wide DNA methylation and the CREB/CREM pathway in the testes of male offspring rats.

This experimental study explored the multigenerational and transgenerational effects of cadmium (Cd) exposure during pregnancy on the testicular tissue and spermatogenesis of male offspring rats. CdCl2 at different doses (0, 0.5, 1, 2 mg/kg/day) were dispensed to pregnant SD rats, thus producing generation F1. Adult females in F1 (PND 56) were mated with untreated fertile males so as to produce generation F2. Likewise, adult females in F2 were mated to produce generation F3. Damages to testicular tissue were observed in all the three generations, with serum testosterone (T) increased in F2 and F3. Notably, the genome-wide DNA methylation level in the testicular tissue of F1 was altered, as was the expression of F1-F3 methyltransferases. In addition, the expression of Creb/Crem pathway, a pathway critical for the metamorphosis from postmeiotic round spermatocytes to spermatozoa, was also remarkably altered in the three generations. In concludion, prenatal Cd exposure might bring multigenerational and transgenerational toxic effects to testes via genome-wide DNA methylation and the regulation of CREB/CREM pathway.

The Wnt/ß-catenin pathway is involved in 2,5-hexanedione-induced ovarian granulosa cell cycle arrest.

N-Hexane causes significant ovarian toxicity, and its main active metabolite 2,5-hexanedione (2,5-HD) can induce ovarian injury through mechanisms such as inducing apoptosis in ovarian granulosa cells (GCs); however, the specific mechanism has not been fully elucidated. In this study, we investigated the effects on the cell cycle of rat ovarian GCs exposed in vitro to different concentrations of 2,5-HD (0 mM, 20 mM, 40 mM, and 60 mM) and further explored the mechanism by mRNA and miRNA microarray analyses. The flow cytometry results sindicated that compared with control cells, in ovarian GCs, there was significant cell cycle arrest after 2,5-HD treatment. Cell cycle- and apoptosis- related gene (Cdk2, Ccnd1, Bax, Bcl-2, Caspase3, and Caspase9) expression was altered. The mRNA and miRNA microarray results suggested that 5678 mRNAs and 32 miRNAs were differentially expressed in the 2,5-HD-treated group. A total of 262 target mRNAs were obtained by miRNA and mRNA coexpression analysis, forming 368 miRNA-mRNA coexpression relationship pairs with 27 miRNAs. GO and KEGG analyses showed that differentially expressed genes were significantly enriched in the cell cycle and Wnt signaling pathways. Furthermore, significant changes in the expression of Wnt signaling pathway and cell cycle- related genes (Fzd1, Lrp6, Tcf3, Tcf4, Fzd6, Lrp5, ß-catenin, Lef1, GSK3ß, and Dvl3) after 2,5-HD treatment were confirmed by qRT-PCR and Western blotting. Ther results of dual-luciferase assays indicated decreased ß-catenin/TCF transcriptional activity after 2,5-HD treatment. In addition, Wnt pathway-related miRNAs (rno-miR-145-5p, rno-miR-143-3p, rno-miR-214-3p, rno-miR-138-5p, and rno-miR-199a-3p) were changed significantly after 2,5-HD treatment. In summary, 2,5-HD induced cell cycle arrest in ovarian GCs, and the Wnt/ß-catenin signaling pathway may play a very critical role in this process. Alterations in the expression of miRNAs such as rno-miR-145-5p may have significant implications.

Green preparation of ß-chitins from squid pens by using alkaline deep eutectic solvents.

Based on the assumption that protein could be removed by the combined mechanism of alkaline induced degradation and strong hydrogen bond interactions of deep eutectic solvents (DESs), ß-chitins were successfully prepared from squid pens by using alkaline DESs formed by potassium carbonate and glycerol. The chemical structures of the DESs were investigated by 1H nuclear magnetic resonance (1H NMR), attenuated total reflection Fourier transform infrared (ATR-FTIR) and molecular modeling, and the physicochemical property of the prepared ß-chitins were characterized. The preparation yields was about 32 %, and DESs with K2CO3/glycerol of 1/10 could be reused for three times while maintaining high preparation yields (31 %-32 %) and degree of deacetylation of 66.9 %-76.9 %. The mechanisms of deproteinization and demineralization by the alkaline DESs were proposed to follow the degradation and dissolution steps, and proteins and minerals were removed from squid pens through the synergistic actions of alkaline degradation and hydrogen bonding interactions. This alkaline DESs are promising to be used as a green and efficient approach for commercial production of ß-chitin.

Single-shot technique of cryoablation for atrial fibrillation has comparable effective and safety outcomes compared to standard technique: insights from multiple clinical studies.

Background: Although there are many freezing protocols available, the optimal freezing dose is still not determined. We aimed to evaluate the effectiveness and safety of different freeze strategies of CBA in the treatment of AF. Methods: PubMed, Cochrane Library, Web of Science, and Embase were searched up to 1st December 2022. Studies comparing the outcomes between single-shot technique and standard technique of cryoablation were included. Subgroup analysis identified potential determinants for single-shot technique procedure. Results: Our search resulted in 3407 records after deduplication. A total of 17 qualified studies met our inclusion criteria. Compared with standard technique, single-shot technique of cryoablation has a comparable rate of freedom from AF/AT(RR 1.00; P = 0.968), a trend for lower rate of procedure complications (RR 0.80; P = 0.069), a lower rate in transient phrenic paralysis (t-PNP) (RR 0.67; P = 0.038), a similar rate in persistent phrenic paralysis (per-PNP) (RR 1.15; P = 0.645), as well as a comparable procedure parameters. Importantly, potentially significant treatment covariable interactions in procedure complications were found in freeze strategy subgroup, male proportion subgroup and age subgroup, including single-shot freeze (RR 1.02; P = 0.915) and TTI-guided (RR 0.63; P = 0.007) with interaction P = 0.051, high male proportion (RR 0.54; P = 0.005) and a low male proportion (RR 0.94; P = 0.759) with interaction P = 0.074, as well as age ≥ 65 (RR0.91; P = 0.642) and age <65 (RR 0.54; P = 0.006),interaction P = 0.090. Meanwhile, only one significant treatment covariable interactions in procedure complications was found in the hypertension subgroup, including HT > 60% (RR 0.89; P = 0.549) and HT ≤ 60% (RR 0. 46; P < 0.01) with interaction P = 0.043. Conclusions: Our study suggested that single-shot technique of cryoablation has comparable effective and safety outcomes for AF ablation compared to standard technique.

Deep Eutectic Solvents Based on L-Arginine and 2-Hydroxypropyl-ß-Cyclodextrin for Drug Carrier and Penetration Enhancement.

By selecting L-arginine as the hydrogen bond acceptor (HBA) and 2-hydroxypropyl-ß-cyclodextrin (2HPßCD) as the hydrogen bond donor (HBD), deep eutectic solvents (DESs) with various water content were prepared at the 4:1 mass ratio of L-arginine to 2HPßCD with 40 to 60% of water, and were studied for its application in transdermal drug delivery system (TDDS). The hydrogen bond networks and internal chemistry structures of the DESs were measured by attenuated total reflection Fourier transform infrared (ATR-FTIR) and 1H-nuclear magnetic resonance spectroscopy (1H-NMR), which demonstrated the successful synthesis of DESs. The viscosity of DES was decreased from 10,324.9 to 3219.6 mPa s, while glass transition temperature (Tg) of the DESs was increased from - 60.8 to - 51.4 °C, as the added water was increased from 45 to 60%. The solubility of ibuprofen, norfloxacin, and nateglinide in DES with 45% of water were increased by 79.3, 44.1, and 3.2 times higher than that in water, respectively. The vitro study of transdermal absorption of lidocaine in DESs showed that the cumulative amounts of lidocaine reached 252.4 µg/cm2, 226.1 µg/cm2, and 286.1 µg/cm2 at 8 h for DESs with 45%, 50%, and 60% of water, respectively. The permeation mechanism of DES with lower content of water (45%) was mainly by changing the fluidization of lipids, while changing the secondary structure of keratin in stratum corneum (SC) at higher water content (50% and 60%). These nonirritant and viscous fluid like DESs with good drug solubility and permeation enhancing effects have broad application prospect in the field of drug solubilization and transdermal drug delivery system.

Prenatal cadmium exposure has inter-generational adverse effects on Sertoli cells through the follicle-stimulating hormone receptor pathway.

In brief: Exposure to cadmium (Cd) during pregnancy can potentially harm the reproductive system of male offspring. This article shows that pregnant woman should be protected from cadmium exposure. Abstract: Exposure to cadmium (Cd) during pregnancy can potentially harm the reproductive system of male offspring, although the full extent of its heritable effects remains partially unresolved. In this study, we examined the inter-generational impacts of Cd using a distinct male-lineage generational model. Pregnant Sprague-Dawley female rats (F0) were administered control or cadmium chloride (0.5, 1 and 2 mg/day) via intra-gastric administration from gestation day 1 to 20. Subsequently, the first filial generation (F1) male rats were mated with untreated females (not exposed to Cd) to produce the second filial generation (F2). Histopathological analysis of the F1 and F2 generations revealed abnormal testicular development, while ultrastructural examination indicated damage to Sertoli cells. Cd exposure also led to alterations in serum hormone levels (gonadotropin-releasing hormone, follicle-stimulating hormone) and reduced follicle-stimulating hormone receptor (FSHR) protein expression in Sertoli cells in the F1 generation. Furthermore, Cd affected the mRNA and protein expression of FSHR pathway factors and DNA methyltransferase, albeit with distinct patterns and inconsistencies observed between the F1 and F2 generations. Overall, our findings indicate that prenatal Cd exposure, using a male-lineage transmission model, can induce inter-generational effects on male reproduction, particularly by causing toxicity in Sertoli cells. This effect appears to be primarily mediated through disruptions in the FSHR pathway and changes in DNA methyltransferase activity in the male testes.

The Hippo signaling pathway contributes to the 2,5-Hexadion-induced apoptosis of ovarian granulosa cells.

Although n-hexane can induce ovarian damage by inducing ovarian granulosa cell (GC) apoptosis, the mechanism underlying this induction of apoptosis has not been fully elucidated. In this study, rat ovarian GCs were exposed to different concentrations of 2,5-hexanedione (2,5-HD) (the main metabolite of n-hexane) in vitro to observe apoptosis, and the mechanism was further explored via mRNA microarray analysis. Hoechst 33258 staining and flow cytometry suggested that the apoptosis rate of ovarian GC apoptosis was significantly increased in the 2,5-HD-treated group. Subsequently, microarray analysis revealed that a total of 5677 mRNAs were differentially expressed, and further GO and KEGG analyses revealed that the differentially expressed genes were significantly enriched in many signaling pathways, including the Hippo pathway. A total of 7 differentially expressed genes that function upstream of the Hippo signaling pathway (Nf2, Wwc1, Ajuba, Llgl1, Dlg3, Rassf6 and Rassf1) were selected to confirm the microarray results by qRT-PCR, and the expression of these genes did change. Subsequently, the expression of key effector genes (Yap1, Mst1 and Lats1) and target genes (Ctgf and Puma) of the Hippo signaling was measured, and the results suggested that the mRNA and protein levels of Yap1, Mst1, Lats1, and Ctgf were significantly decreased while those of Puma were significantly increased after 2,5-HD treatment. Further CO-IP analysis suggested that the interaction between YAP1 and TEAD was significantly reduced after 2,5-HD treatment, while the interaction between YAP1 and P73 was not affected. In summary, during the 2,5-HD-induced apoptosis of ovarian GCs, the Hippo signaling pathway is inhibited, and downregulation of the pro-proliferation gene Ctgf and upregulated of the pro-apoptosis gene Puma are important. Decreased Ctgf expression was associated with decreased binding of YAP1 to TEAD. However, increased PUMA expression was not associated with YAP1 binding to P73.

Maternal genetic intergenerational and transgenerational effects on hormone synthesis in ovarian granulosa cells of offspring exposed to cadmium during pregnancy.

This study aimed to investigate the maternally inherited intergenerational and transgenerational effects of cadmium (Cd) exposure on steroid hormone synthesis in the ovarian granulosa cells (GCs) of offspring rats. F1 rats were obtained by mating adult female Sprague-Dawley rats with healthy adult male rats and were exposed to 0, 0.5, 2.0, and 8.0 mg/kg CdCl2 during pregnancy. The adult female rats (PND 56) were mated with healthy adult male rats to produce F2 and F3 rats. The serum progesterone (Pg) and estradiol (E2) levels of the F2 adult female rats were decreased, while those of F3 rats were significantly increased. Moreover, hormone synthesis-related genes had different expression patterns in the F2 and F3 generations. F2 and F3 rat ovarian GCs exhibited altered miRNA expression profiles and DNA methylation patterns. Validation of miRNAs that regulate hormone synthesis-related genes in the cAMP/PKA signaling pathway suggested that miR-124-3p was downregulated in F2 and F3 rats, while miR-133a-5p and miR-150-5p were upregulated in F2 rats and downregulated in F3 rats. In summary, 1) there are maternal genetic intergenerational (GCs hormone synthesis disorder) and transgenerational (GCs hormone synthesis function repair change) effects on hormone synthesis function changes in offspring GCs induced by Cd exposure during pregnancy. 2) Changes in miRNAs and DNA methylation modifications associated with the genetic effects of altered hormone synthesis function in offspring GCs induced by Cd exposure during pregnancy are important. 3) Under the current environmental level of Cd exposure, the possible risk of maternal genetic intergenerational and transgenerational effects of offspring ovarian toxicity should be strongly considered.