Ionizable Polymeric Micelles with Phenylalanine Moieties Enhance Intracellular Delivery of Self-Replicating RNA for Long-Lasting Protein Expression In Vivo.

mRNA-based therapeutics are revolutionizing the landscape of medical interventions. However, the short half-life of mRNA and transient protein expression often limits its therapeutic potential, demanding high treatment doses or repeated administrations. Self-replicating RNA (RepRNA)-based treatments could offer enhanced protein production and reduce the required dosage. Here, we developed polymeric micelles based on flexible poly(ethylene glycol)-poly(glycerol) (PEG-PG) block copolymers modified with phenylalanine (Phe) moieties via biodegradable ester bonds for the efficient delivery of RepRNA. These polymers successfully encapsulated RepRNA into sub-100 nm micelles assisted by the hydrophobicity of the Phe moieties and their ability to π-π stack with the bases in RepRNA. The micelles made from Phe-modified PEG-PG (PEG-PG(Phe)) effectively maintained the integrity of the loaded RepRNA in RNase-rich serum conditions. Once taken up by cells, the micelles triggered a pH-responsive membrane disruption, promoted by the strong protonation of the amino groups at endosomal pH, thereby delivering the RepRNA to the cytosol. The system induced strong protein expression in vitro and outperformed commercial transfecting reagents in vivo, where it resulted in enhanced and long-lasting protein expression.

Identification and Functional Analysis of a Defensin CcDef2 from Coridius chinensis.

Coridius chinensis belongs to Dinidoridae, Hemiptera. Previous studies have indicated that C. chinensis contains abundant polypeptides with antibacterial and anticancer activities. Antimicrobial peptides (AMPs), as endogenous peptides with immune function, play an indispensable role in the process of biological development and immunity. AMPs have become one of the most potential substitutes for antibiotics due to their small molecular weight and broad-spectrum antimicrobial activity. In this study, a defensin CcDef2 from C. chinensis was characterized based on bioinformatics and functional analyses. The mature peptide of CcDef2 is a typical cationic peptide composed of 43 amino acid residues with five cations, and contains three intramolecular disulfide bonds and a typical cysteine-stabilized αß motif in defensins. Phylogenetic analysis showed that CcDef2 belongs to the insect defensin family. Analysis of gene expression patterns showed that CcDef2 was expressed throughout developmental stages of C. chinensis with high levels at the nymphal stage and in adult tissues tested with the highest level in the fat body. In addition, the CcDef2 expression was significantly upregulated in adults infected by bacteria. After expressed in Escherichia coli BL21(DE3) and renatured, the recombinant CcDef2 showed a significant antibacterial effect on three kinds of Gram-positive bacteria. These results indicate that CcDef2 is an excellent antibacterial peptide and a highly effective immune effector in the innate immunity of C. chinensis. This study provides a foundation for further understanding the function of CcDef2 and developing new antimicrobial drugs.

[Expression of HSP70 in Kunming Mouse Embryos Stimulated by Chronic Mild Anticipatory Stress].

OBJECTIVE: To measure the expression of heat shock protein 70 (HSP70) in mice embryos with the stimulation of chronic mild anticipatory stress (CUMS) to female Kunming mice. METHODS: Three hundreds female Kunming mice were stressed by 9 chronic mild unpredictable stress factors for 28 days and then divided into three groups of mild, moderate and severe stress. PMSG/hCG was measured to assess the induction of superovulation, and ovarian response and embryo development potential were observed. The expression of HSP70 in 2-cell embryos and day 4 embryos was detected by immunofluorescence and real time polymerase chain reaction (RT-PCR). RESULTS: After 28 d CUMS stimulation, the rate of mice in mild, moderate and severe stress were 50%, 32% and 18%, respectively. In the mild stress group, ovarian response and oocyte development potential were similar to those of control, while HSP70 expression of the embryos was significantly higher (P<0.05). In the severe stress group, ovarian response and oocyte development potential were significantly decreased compared with the control group (P<0.05), while HSP70 expression was similar to that of control. CONCLUSION: HSP70 overexpression observed in the embryos may be related to its proteetive effect against chronic unpredictable stress.

bFGF and VEGF improve the quality of vitrified-thawed human ovarian tissues after xenotransplantation to SCID mice.

PURPOSE: The aim of this research is to study whether basic fibroblast growth factor (bFGF) alone or in combination with vascular endothelial growth factor (VEGF) could improve the quality of vitrified-thawed human ovarian tissue xenotransplanted to severe combined immune deficiency (SCID) mice. METHODS: After collection and cryopreservation, thawed human ovarian tissue were cultured in vitro for 2 days and then xenografted to severe combined immune deficiency (SCID) mice for 7 days. The in vitro culture medium was separated into six groups, including (A) the blank control group, (B) the human recombinant bFGF (150 ng/ml) group, (C) the bFGF (150 ng/ml)+human recombinant VEGF (25 ng/ml) group, (D) bFGF (150 ng/ml)+VEGF (50 ng/ml) group, (E) bFGF (150 ng/ml)+ VEGF (75 ng/ml) group and (F) bFGF (150 ng/ml) + VEGF (100 ng/ml) group. In addition, eight pieces of thawed ovarian tissue were transplanted without in vitro culture, which serve as the fresh control group. The effect of transplantation was assessed by histological analysis, immunohistochemical staining for CD34, Ki-67, and AC-3 expression, and microvessel density (MVD). RESULTS: There was no significant difference between the fresh and blank control group. Compared to the blank control group, the number of follicles, MVD, and rate of Ki-67-positive cells increased significantly in groups B, C, D, E, and F, while apoptosis decreased significantly. Compared to the bFGF treatment group, no significant difference appeared in group C, D, E, and F. CONCLUSIONS: The administration of bFGF alone or in combination with VEGF improved the quality of postgraft human ovarian tissue, though VEGF, regardless of different concentrations, did not influence effect of bFGF.

HSP70 overexpression may play a protective role in the mouse embryos stimulated by CUMS.

BACKGROUND: We evaluated whether heat shock protein HSP70 plays a protective role in the embryos of Kunming mice subjected to chronic unpredictable mild stress. METHODS: Female mice were stimulated for 4 weeks with nine stressors and then divided into mild, moderate and severe stress groups. Superovulation was induced with a gonadotropin preparation (PMSG/HCG) and HSP70 expression in 2-cell embryos and day 4 embryos was detected by immunofluorescence (IF) and real-time polymerase chain reaction (RT-PCR). RESULTS: In the mild stress group, ovarian response and oocyte development potential were similar to those of the control group, while the HSP70 mRNA levels of the embryos were significantly higher (P < 0.05). In the severe stress group, ovarian response and oocyte development potential decreased compared with the control group (P < 0.05), while the HSP70 mRNA levels were similar. The results of the moderate stress group were intermediate among the three groups. Furthermore, HSP70 mRNA levels of the embryos were shown to be positively associated with parameters of oocyte and embryo development potential (P < 0.05). CONCLUSIONS: HSP70 overexpression may play a protective role in the embryos of the mild or moderate stress mice stimulated by chronic unpredictable mild stress.

CYP1B1 C4326G polymorphism and susceptibility to cervical cancer in Chinese Han women.

Cytochrome P450 1B1 (CYP1B1) is a key P450 enzyme, which could catalyze the formation of 4-hydroxy estrogen metabolites and play a role in estrogen-dependent cancers. We hypothesized that genetic variant in CYP1B1 may modify individual susceptibility to cervical cancer. The aim of this study was to evaluate the association between CYP1B1 C4326G polymorphism and cervical cancer risk in Chinese women. We extracted the peripheral blood samples in 250 patients with cervical cancer and 250 female controls. The matrix-assisted laser desorption ionization time-of-flight mass spectrometry method and direct DNA sequencing were performed to detect the polymorphism. The frequencies of CC, CG, and GG genotypes of CYP1B1 C4326G in cases and controls were 66.0, 26.8, 7.2% and 75.2, 21.6, and 3.2%, respectively, and there was a significant difference between the two groups (P = 0.034). Compared with the wild-type CC genotype, the variant GG genotype was associated with a significantly increased risk of cervical cancer (adjusted OR = 2.30; 95% CI = 1.02, 5.50). Moreover, stratification analysis by age, smoking, drinking, human papillomaviruses (HPV) 16 or 18 carrier status, and family history of cervical cancer, we found that the variant genotypes containing the G allele were associated with a significantly increased risk of cervical cancer among HPV 16 or 18-positive individuals (adjusted OR = 2.85; 95% CI = 1.45, 5.62) and among women younger than 45 years old (adjusted OR = 1.87; 95% CI = 1.03, 3.37). These results suggest that CYP1B1 C4326G polymorphism may increase risk of cervical cancer in Chinese women, especially among young individuals with high-risk HPV infection.

In vitro culture thawed human ovarian tissue: NIV versus slow freezing method.

The aim of this study was to determine if the needle immersed vitrification method (NIV) can improve the growth potential of thawed ovarian tissue in vitro culture. Human ovarian cortical tissues were cryopreserved using NIV and slow freezing method. After 14 days of culture, the preservation outcomes of NIV and slow freezing groups were analyzed histologically using light microscope and apoptosis was assessed by TUNEL assay. The result showed that the percentage of morphologically abnormal primordial follicles was lower in NIV group than in slow freezing group (P < 0.05). The incidence of TUNEL-positive primordial follicles was lower in NIV group than in slow freezing group (P < 0.05). The study showed that cryopreservation of human ovarian tissue with NIV was effective in improving the growth potential of frozen-thawed ovarian tissue in vitro culture.

An update on renal fibrosis: from mechanisms to therapeutic strategies with a focus on extracellular vesicles.

The increasing prevalence of chronic kidney disease (CKD) is a major global public health concern. Despite the complicated pathogenesis of CKD, renal fibrosis represents the most common pathological condition, comprised of progressive accumulation of extracellular matrix in the diseased kidney. Over the last several decades, tremendous progress in understanding the mechanism of renal fibrosis has been achieved, and corresponding potential therapeutic strategies targeting fibrosis-related signaling pathways are emerging. Importantly, extracellular vesicles (EVs) contribute significantly to renal inflammation and fibrosis by mediating cellular communication. Increasing evidence suggests the potential of EV-based therapy in renal inflammation and fibrosis, which may represent a future direction for CKD therapy.

FOXO1 expression and regulation in endometrial tissue during the menstrual cycle and in early pregnancy decidua.

AIMS: To determine FOXO1 (forkhead box O1) expression in the human endometrium during the menstrual cycle and decidua of early pregnancy, as well as FOXO1 regulation in endometrial stromal cells (ESC). METHODS: Immunohistochemistry, RT-qPCR, and Western blot analyses evaluated cellular localization and altered FOXO1 expression in the endometrium during the menstrual cycle and decidua of early pregnancy (proliferative phase, n = 12; early-secretory phase, n = 7; mid-secretory phase, n = 10; late-secretory phase, n = 10; early pregnancy, n = 12). Using RT-qPCR and Western blot, we studied the regulation of FOXO1 by 8-bromo-cAMP, 4-pregnene-3,20-dione, 17ß-estradiol, and human chorionic gonadotrophin in ESC (n = 5). RESULTS: The expression level of FOXO1 in human endometrial tissue fluctuated with the menstrual cycle. If pregnancy occurred, the expression of FOXO1 was further increased (p < 0.05) and cAMP regulated FOXO1 expression in ESC. In addition, 4-pregnene-3,20-dione cooperatively stimulated FOXO1 expression with cAMP. We also observed FOXO1 expression during in vitro cAMP-induced decidualization. CONCLUSIONS: The pattern of FOXO1 expression suggests a potential role for FOXO1 in implantation and decidualization.

Outcome of conventional IVF and ICSI on sibling oocytes in the case of isolated teratozoospermia.

PURPOSE: To reevaluate the effect of isolated teratozoospermia on IVF and determine if there was any therapeutic benefit to isolated teratozoospermia by ICSI, since there are no widely accepted criteria for the treatment technique about isolated teratozoospermia. METHODS: A total of 441 couples with >20 million and progressive motility >30 % sperm undergoing their first IVF/ICSI cycle were included in the study between 2008 and 2010, for whom at least 8 oocytes were retrived. Isolated teratozoospermia was diagnosed in 183 of the included couples, and the rest couples (normal sperm morphology) were studied as control. Sibling oocytes were randomized to be inseminated either by ICSI or IVF. Fertilization rate, embryo quality, pregnancy rate, implantation rate and spontaneous abortion rate were assessed. RESULTS: There was no difference in the percentage of eggs fertilized, implantation rate, pregnancy rate and spontaneous abortion rate between conventional IVF and ICSI regardless of the percentage of normal morphology. The day 3 embryonic morphology and rate of development were not different despite the insemination method and percentage of normal morphology. CONCLUSION: Because isolated teratozoospermia did not influence the major indices of IVF and the unnecessary use of ICSI is time-consuming, costly and potential risks, couples with isolated teratozoospermia need not be subjected to ICSI.

Glucosamine-6-phosphate N-acetyltransferase gene silencing by parental RNA interference in rice leaf folder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae).

Parental RNAi (pRNAi) is a response of RNA interference in which treated insect pests progenies showed a gene silencing phenotypes. pRNAi of CmGNA gene has been studied in Cnaphalocrocis medinalis via injection. Our results showed significant reduction in ovulation per female that was 26% and 35.26% in G1 and G2 generations, respectively. Significant reduction of hatched eggs per female were observed 23.53% and 45.26% as compared to control in G1-G2 generations, respectively. We also observed the significant variation in the sex ratio between female (40% and 53%) in G1-G2 generations, and in male (65%) in G1 generation as compared to control. Our results also demonstrated the significant larval mortality (63% and 55%) and pupal mortality (55% and 41%), and significant reduction of mRNA expression level in G1 and G2 generations. Our findings have confirmed that effectiveness of pRNAi induced silencing on the CmGNA target gene in G1-G2 generations of C. medinalis. These results suggested the potential role of pRNAi in insect pest resistance management strategies.

Dietary Methylglyoxal Exposure Induces Alzheimer's Disease by Promoting Amyloid ß Accumulation and Disrupting Autophagy in Caenorhabditis elegans.

Methylglyoxal (MG) is a precursor of advanced glycation end products usually generated during cooking. The high level of MG in the brain is correlated to the pathogenesis of Alzheimer's disease (AD). However, it is not clear if MG consumed through the diet can cause AD-related toxicity. Herein, the Caenorhabditis elegans (C. elegans) AD model was used to investigate the neurotoxicity after long-term MG exposure at dietary levels. The results showed that C. elegans locomotive behaviors were significantly decreased after 0.1, 0.5, and 1 mM MG exposure (p < 0.001). In amyloid ß (Aß)-expressing transgenic C. elegans strains, 0.5 mM MG significantly promoted Aß accumulation by around 50% in day-8 CL2006 (p < 0.001), enhanced paralysis in CL4176 (p < 0.001) and CL2006 (p < 0.01), and made CL2355 around 17% more vulnerable to 5-HT, indicating impaired serotonin reuptake (p < 0.05). Additionally, 0.5 mM MG significantly increased the reactive oxygen species level (p < 0.001) by inhibiting the expression of stress-response genes including sod-3, gst-4, and hsp-16.2 in day-8 aged worms. Moreover, the autophagic pathway was disrupted through lgg-1, vps-34, and bec-1 expression after MG exposure and Aß accumulation. Treatment with the citrus flavonoid nobiletin reduced the MG-induced toxicity (p < 0.001). Overall, these findings imply that it is possible to exacerbate AD pathogenesis by MG exposure through the diet.

Influence of the Wenchuan earthquake on self-reported irregular menstrual cycles in surviving women.

OBJECTIVE: To explore the influence of stress induced by the Wenchuan earthquake on the menstrual cycles of surviving women. METHODS: Self-reports of the menstrual cycles of 473 women that survived the Wenchuan earthquake were analyzed. Menstrual regularity was defined as menses between 21 and 35 days long. The death of a child or the loss of property and social resources was verified for all surviving women. The severity of these losses was assessed and graded as high, little, and none. RESULTS: About 21% of the study participants reported that their menstrual cycles became irregular after the Wenchuan earthquake, and this percentage was significantly higher than before the earthquake (6%, p < 0.05). About 30% of the surviving women with a high degree of loss in the earthquake reported menstrual irregularity after the earthquake. Association analyses showed that some stressors of the Wenchuan earthquake were strongly associated with self-reports of menstrual irregularity, including the loss of children (RR: 1.58; 95% CI: 1.09, 2.28), large amounts of property (RR: 1.49; 95% CI: 1.03, 2.15), social resources (RR: 1.34; 95% CI: 1.00, 1.80) and the hormonal contraception use (RR: 1.62; 95% CI: 1.21, 1.83). CONCLUSIONS: Self-reported menstrual irregularity is common in women that survived the Wenchuan earthquake, especially in those who lost children, large amounts of property and social resources.

Baseline psychological stress and ovarian norepinephrine levels negatively affect the outcome of in vitro fertilisation.

OBJECTIVE: We determined the effect of baseline psychological stress and norepinephrine (NE) levels in the follicular fluid on the outcome of in vitro fertilisation (IVF). METHODS: One hundred seven women with tubal factor infertility were evaluated before and during their first IVF treatment. On the first day of down-regulation, their psychological state was measured using the Zung Self-rating Anxiety (SAS) and Depression Scales (SDS), and defined as baseline psychological stress. On oocyte retrieval day, NE levels in the follicular fluid and peripheral plasma were measured by high-performance liquid chromatography. RESULTS: On the first day of down-regulation, nearly 10% of the women with tubal factor infertility reported that they were experiencing both anxiety and depression. This baseline psychological stress was lower in pregnant (3.7%) than in non-pregnant women (15.7%, p<0.05). On oocyte retrieval day, the NE levels in follicular fluid were almost twice the amount seen in peripheral plasma, but did not differ in pregnant and non-pregnant women (p>0.05). NE levels in follicular fluid were negatively associated with the percentage of good quality embryos (r= -0.62, p<0.05). CONCLUSIONS: Baseline psychological stress (both anxiety and depression) may negatively influence the clinical pregnancy rate of IVF treatment in women with tubal factor infertility. Furthermore, follicular NE levels are negatively associated with the percentage of good quality embryos.

Identification and Functional Analysis of a Lysozyme Gene from Coridius chinensis (Hemiptera: Dinidoridae).

Coridius chinensis is a valuable medicinal insect resource in China. Previous studies have indicated that the antibacterial and anticancer effects of the C. chinensis extract mainly come from the active polypeptides. Lysozyme is an effective immune effector in insect innate immunity and usually has excellent bactericidal effects. There are two kinds of lysozymes in insects, c-type and i-type, which play an important role in innate immunity and intestinal digestion. Studying lysozyme in C. chinensis will be helpful to further explore the evolutionary relationship and functional differences among lysozymes of various species and to determine whether they have biological activity and medicinal value. In this study, a lysozyme CcLys2 was identified from C. chinensis. CcLys2 contains 223 amino acid residues, and possesses a typical domain of the c-type lysozyme and a putative catalytic site formed by two conserved residues Glu32 and Asp50. Phylogenetic analysis showed that CcLys2 belongs to the H-branch of the c-type lysozyme. The analysis of spatiotemporal expression patterns indicated that CcLys2 was mainly expressed in the fat body of C. chinensis adults and was highly expressed in the second- and fifth-instar nymphs. In addition, CcLys2 was significantly up-regulated after injecting and feeding bacteria. In the bacterial inhibition assay, it was found that CcLys2 had antibacterial activity against Gram-positive bacteria at a low pH. These results indicate that CcLys2 has muramidase activity, involves in the innate immunity of C. chinensis, and is also closely related to the bacterial immune defense or digestive function of the intestine.

Cloning, Characterization, and RNA Interference Effect of the UDP-N-Acetylglucosamine Pyrophosphorylase Gene in Cnaphalocrocis medinalis.

The rice leaf folder, Cnaphalocrocis medinalis is a major pest of rice and is difficult to control. UDP-N-acetylglucosamine pyrophosphorylase (UAP) is a key enzyme in the chitin synthesis pathway in insects. In this study, the UAP gene from C. medinalis (CmUAP) was cloned and characterized. The cDNA of CmUAP is 1788 bp in length, containing an open reading frame of 1464 nucleotides that encodes 487 amino acids. Homology and phylogenetic analyses of the predicted protein indicated that CmUAP shared 91.79%, 87.89%, and 82.75% identities with UAPs of Glyphodes pyloalis, Ostrinia furnacalis, and Heortia vitessoides, respectively. Expression pattern analyses by droplet digital PCR demonstrated that CmUAP was expressed at all developmental stages and in 12 tissues of C. medinalis adults. Silencing of CmUAP by injection of double-stranded RNA specific to CmUAP caused death, slow growth, reduced feeding and excretion, and weight loss in C. medinalis larvae; meanwhile, severe developmental disorders were observed. The findings suggest that CmUAP is essential for the growth and development of C. medinalis, and that targeting the CmUAP gene through RNAi technology can be used for biological control of this insect.

Cryopreservation of the human ovarian tissue induces the expression of Fas system in morphologically normal primordial follicles.

The aim of this study was to investigate whether the Fas system was involved in cryopreservation process of the human ovarian tissue. Human ovarian cortical tissues were cryopreserved using slow freezing method. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was used to investigate apoptosis of primordial follicle. Fas, Fas ligand and active caspase-3 expression were detected by means of immunohistochemistry. The results showed that immunostaining for Fas, Fas ligand and active caspase-3 were not detected in morphologically normal primordial follicles in fresh ovarian tissue. After cryopreservation, Fas, Fas ligand and active caspase-3 immunostaining were present in morphologically normal primordial follicles. The study showed that cryopreservation of the human ovarian tissue induced the expression of Fas system in morphologically normal primordial follicles.

Effectiveness of bromocriptine monotherapy or combination treatment with clomiphene for infertility in women with galactorrhea and normal prolactin: A systematic review and meta-analysis.

BACKGROUND: Among women with unexplained infertility, 28% to 55% of patients with galactorrhea are normoprolactinemic. Bromocriptine, a common treatment for infertile women with hyperprolactinemia, has been used in the treatment of unexplained subfertility in women with galactorrhea and normal prolactin; however, its effectiveness and safety profile have never been determined. OBJECTIVE: The aim of this study was to determine the relative effectiveness and safety profile of bromocriptine monotherapy or as an adjunct to clomiphene citrate in women with galactorrhea and normal prolactin levels. METHODS: We conducted a search of the Cochrane Subfertility Review Group specialized register of controlled trials (March 2010), CENTRAL (The Cochrane Library, Issue 3, 2010), MEDLINE (1950-March 2010), EMBASE (1980-March 2010), and the China Biological Medicine Database (inception to March 2010) for relevant randomized controlled trials (RCTs) using the following terms: controlled, randomized, blinded, clinical trials, humans, galactorrhea, prolactin, bromocriptine, infertility, and subfertility. Additionally, reference lists of identified articles were searched for relevant articles. RESULTS: Of the 8 studies identified, 5 were excluded after full-text review for the following reasons: lack of a placebo group (2); difference in cointerventions (1); difference in end points (1); and systematic review (1). Therefore, 3 RCTs were included in this review. Bromocriptine administered in combination with clomiphene was found to be associated with a higher accumulative pregnancy rate compared with clomiphene monotherapy (fixed odds ratio [OR], 5.33; 95% CI, 2.62-10.88), and a lower miscarriage rate (fixed OR, 0.20; 95% CI, 0.05-0.76). Only 1 trial reported live birth as an outcome, and multiple pregnancy rates were poorly reported. Patient-reported adverse effects were mentioned in the studies, but reports were often incomplete. CONCLUSIONS: This review suggests the effectiveness of bromocriptine with clomiphene for infertility in women with galactorrhea and normal prolactin levels. Further RCTs of adequate power and of high methodologic quality are required to confirm these findings.

[Effect of TGF-beta1 on embryo implantation and development in mice in vitro].

OBJECTIVE: To investigate the role of TGF-beta1 in embryo implantation and development in vitro in mice. METHODS: Mouse embryos at 2-cell stage were cultured in the media of M16 with exposure to different levels of TGF-beta1 (0, 1, 10 and 50 ng/mL). The percentage of embryos reaching fixed stages (early blastocyst, expanding blastocyst and hatched blastocyst) was monitored 68 h and 92 h after the culture. The expanding blastocys cultured for 68 h in M16 without TGF-beta1 and those with 10 ng/mL of TGF-beta1 were transferred to pseudopregnant mice. On the 6th day post transfer, the successful rates of implantation were counted. The level of IL-10/IFN-gamma in the serum and maternal-fetus interface of the mice was detected by ELISA on the 6th day post transfer. RESULTS: TGF-beta1 improved embryo growth in vitro. TGF-beta1 at a level of 10 ng/mL had the maximum impact, with 15.6%, 68.09%, 1.42% of embryos reaching early, expanding, and hatched stage, respectively, 68 h after culture, and 6.38%, 28.37%, 53.19% of embryos reaching early, expanding, and hatched stage, respectively, 92 h after culture. The promoting effect declined when TGF-beta1 reached 50 ng/mL. The successful rate of implantation of embryos cultured in M16 with TGF-beta1 was significantly higher than those cultured in M16 without TGF-beta1 (35. 2% vs. 17.19%, P < 0.05). The embryos cultured in M16 with TGF-beta1 had significantly lower level of IFN-gamma in the maternal-fetus interface than those cultured in M16 without TGF-beta1 [(30.89 +/- 11.31) pg/mL vs. (43.23 +/- 18. 09) pg/mL, P < 0.053. CONCLUSION: TGF-beta1 at an appropriate dose improves embryo implantation in mice in vitro. The mechanism may involve the improvement of the quality of embryos and their development, and decrease of IFN-gamma synthesis in maternal-fetal interface, a chemical that could cause Th2 bias.

Early-life long-term exposure to ZnO nanoparticles suppresses innate immunity regulated by SKN-1/Nrf and the p38 MAPK signaling pathway in Caenorhabditis elegans.

The widespread use of zinc oxide nanoparticles (ZnO-NPs) has led to their release into the environment, and they thus represent a potential risk for both humans and ecosystems. However, the negative impact of ZnO-NPs on the immune system, especially in relation to host defense against pathogenic infection and its underlying regulatory mechanisms, remains largely unexplored. This study investigated the effects of early-life long-term ZnO-NPs exposure (from L1 larvae to adults) on innate immunity and its underlying mechanisms using a host-pathogen Caenorhabditis elegans model, and this was compared with the effect of ionic Zn. The results showed that the ZnO-NPs taken up by C. elegans primarily accumulated in the intestine and that early-life long-term ZnO-NPs exposure at environmentally relevant concentrations (50 and 500 µg/L) decreased the survival of wild-type C. elegans when faced with pathogenic Pseudomonas aeruginosa PA14 infection. Early-life long-term ZnO-NPs (500 µg/L) exposure significantly increased (by about 3-fold) the accumulation of live P. aeruginosa PA14 colonies in the intestine of C. elegans. In addition, ZnO-NPs (500 µg/L) inhibited the intestinal nuclear translocation of SKN-1 and also downregulated gcs-1 gene expression, which is an SKN-1 target gene. Further evidence revealed that early-life long-term exposure to ZnO-NPs (500 µg/L) did not increase susceptibility to mutation among the genes (pmk-1, sek-1, and nsy-1) encoding the p38 mitogen-activated protein kinase (MAPK) cascade in response to P. aeruginosa PA14 infection, though ZnO-NPs significantly decreased the mRNA levels of pmk-1, sek-1, and nsy-1. This study provides regulatory insight based on evidence that ZnO-NPs suppress the innate immunity of C. elegans and highlights the potential health risks of certain environmental nanomaterials, including ZnO-NPs, in terms of their immunotoxicity at environmentally relevant concentrations.