NSUN2-Mediated m5C Methylation Impairs Endometrial Receptivity.

Impaired endometrial decidualization is the primary cause of recurrent implantation failure (RIF). RNA methylation modification, especially NSUN family mediated m5C, is crucial for various physiological events, such as maternal-to-zygotic transition, gametogenesis, embryonic development, organismal lifespan, and cell cycle. However, the regulatory mechanisms between NSUN family mediated m5C modification and RIF remain unknown. We acquired NSUN2 expression data of 15 human endometrium samples at proliferative and secretory stages from reproductive cell atlas. The overall pattern of m5C sites and genes was elucidated through m5C-BS-seq, whereas the overall m5C levels in different groups were revealed by dot blot assay. BrdU and western blotting assays were carried out to evaluate the role of NSUN2 in proliferation and autophagy. The effects of NSUN2-mediated m5C modification on embryo attachment were evaluated by an in vitro model of a confluent monolayer of Ishikawa cells cocultured with BeWo spheroids, and its downstream targets were evaluated by real-time reverse-transcription PCR and western blotting in Ishikawa cells. The molecular mechanism for NSUN2 regulating its downstream targets' expression was determined by Cut&Tag and coimmunoprecipitation assays. NSUN2 was increased in SOX9+ cells and widespread in epithelial cell type at the proliferative stage by previous single-cell RNA sequencing data. NSUN2 overexpression (NSUN2OE) in the Ishikawa cell line elevated m5C levels and promoted cell proliferation and autophagy. NSUN2OE reduced attachment efficiency of BeWo cell spheres. Overexpressed NSUN2 was found to increase STAT1 and MMP14 mRNA expressions by inducing exon skipping. NSUN2 interacted with CLDN4 through m5C modification, and NSUN2OE or NSUN2 knockdown resulted in a similar variation tendency of CLDN4. Overexpression of NSUN2 increased CLDN4 H3K9ac modification by downregulating SIRT4 expression at the protein level, leading to the upregulation of CLDN4 mRNA expression. Our results uncovered a novel intricate regulatory mechanism between NSUN2-mediated m5C and RIF and suggested a potential new therapeutic strategy for RIF.

Transplantation of olfactory ensheathing cells can alleviate neuroinflammatory responses in rats with trigeminal neuralgia.

Trigeminal neuralgia (TN) is a common form of facial pain, which primarily manifests as severe pain similar to facial acupuncture and electric shock. Olfactory ensheathing cells (OECs) are glial cells with high bioactivity; these cells are essential for the periodic regeneration of the olfactory nerve and have been utilized for the repair of nerve injuries. A member of the P2X receptor family, P2X7R, is an ion channel type receptor that has been confirmed to participate in various pain response processes. In this study, we transplanted OECs into trigeminal nerve-model rats with distal infraorbital nerve ligation to observe the therapeutic effect of transplanted OECs in rats. Additionally, we utilized the P2X7R-specific inhibitor brilliant blue G (BBG) to study the therapeutic mechanisms of cell transplantation. The facial mechanical pain threshold of these rats significantly increased following cell transplantation. The immunohistochemistry, immunoblotting, and RT-qPCR results demonstrated that the levels of P2X7R, (NOD)-like receptor protein-3 (NLRP3), nuclear factor-κB (NF-κB), interleukin (IL)-1ß, and IL-18 in the trigeminal ganglion of rats treated with OEC transplantation or BBG treatment were significantly lower than those in the injured group without treatment. Overall, our results demonstrate that OEC transplantation can alleviate TN in rats, and it can reduce the expression of P2X7R related inflammatory factors in TN rats, reducing neuroinflammatory response in TG.

A low-cost, low-input method establishment for m6A MeRIP-seq.

N6-methyladenosine (m6A) is a highly prevalent modification found in mammal mRNA molecules that plays a crucial role in the regulation of cellular function. m6A RNA immunoprecipitation sequencing (MeRIP-seq) has been frequently used in transcriptomics research to identify the location of m6A. MABE572 (Millipore) is the most widely utilized and efficient anti-m6A antibody for MeRIP-seq. However, due to the high dose and price of this antibody, which has also been taken off the market, we discovered that CST's anti-m6A antibody can be used instead of MABE572 to map the m6A transcriptome. In the present study, we performed different concentrations of the CST anti-m6A antibodies with the corresponding initiation RNA of HEK293T cells, 2.5 µg antibody with 1 µg total RNA, 1.25 µg antibody with 0.5 µg total RNA, and 1.25 µg antibody with 0.1 µg total RNA. By comparing the m6A peak calling, enriched motifs, alternative splicing events, and nuclear transcripts modified by m6A between the CST and Millipore libraries, it was found that the CST library presented similar data to Millipore, even at incredibly low doses. The volume and cost of antibodies are significantly reduced by this refined MeRIP-seq using CST antibody, making it convenient to map future large-scale sample m6A methylation.

ATP ion channel P2X purinergic receptors in inflammation response.

Different studies have confirmed that P2X purinergic receptors play a key role in inflammation. Activation of P2X purinergic receptors can release inflammatory cytokines and participate in the progression of inflammatory diseases. In an inflammatory microenvironment, cells can release a large amount of ATP to activate P2X receptors, open non-selective cation channels, activate multiple intracellular signaling, release multiple inflammatory cytokines, amplify inflammatory response. While P2X4 and P2X7 receptors play an important role in the process of inflammation. P2X4 receptor can mediate the activation of microglia involved in neuroinflammation, and P2X7 receptor can mediate different inflammatory cells to mediate the progression of tissue-wide inflammation. At present, the role of P2X receptors in inflammatory response has been widely recognized and affirmed. Therefore, in this paper, we discussed the role of P2X receptors-mediated inflammation. Moreover, we also described the effects of some antagonists (such as A-438079, 5-BDBD, A-804598, A-839977, and A-740003) on inflammation relief by antagonizing the activities of P2X receptors.

[Pollution Characteristics of PM2.5 Chemical Composition in Zhejiang Province].

To investigate the seasonal and regional pollution characteristics of PM2.5 chemical composition in Zhejiang province, this study was based on manual sampling monitoring data from 11 sampling sites of four regions in Zhejiang province from October 1, 2019 to September 30, 2020. The results showed that during the observation period, the average ρ(PM2.5) of the four regions ranged from 34.3 µg·m-3 to 46.4 µg·m-3. The PM2.5 mass concentrations in the hinterland areas of western Zhejiang and northern Zhejiang were relatively high, 15.1% and 13.2% higher than the mean value, respectively. The PM2.5 mass concentrations in the coastal areas of eastern Zhejiang and southern Zhejiang were relatively low, 8.4% and 14.9% lower than the average, respectively. The seasonal characteristics showed a higher concentration in autumn and winter and lowest concentration in summer. The seasonal variation in PM2.5 mass concentration from autumn to spring was not obvious in southern Zhejiang, whereas in western Zhejiang, the PM2.5 mass concentration followed a descending sequence of autumn>winter>spring>summer. In northern Zhejiang and eastern Zhejiang, the trend was winter>autumn>spring>summer. During the observation period in the inland area, the ρ(PM2.5) of the scenic area, administrative area, residential area, and mixed area of commercial traffic and residents were (40.2±10.2), (46.3±9.6), (50.1±10.6), and (46.7±10.2) µg·m-3, respectively. The highest value of ρ(PM2.5) was in the residential area. During the sampling period in coastal areas, the ρ(PM2.5) of the cultural and entertainment area and mixed area of commercial traffic and residents were (27.4±5.8) µg·m-3and (37.2±5.6) µg·m-3, respectively. The contribution rates of organic matter (OM), NO3-, SO42-, NH4+, trace elements, and crustal matter in PM2.5were 26.4%, 15.4%, 12.4%, 9.0%, 7.1%, and 5.7%, respectively. The SNA, including SO42-, NO3-, and NH4+, contributed 36.8% in PM2.5. In terms of seasons, the contribution of OM to PM2.5 in autumn, spring, and summer was higher than that of other compositions, which accounted for 28.3%, 27.7%, and 26.3%, respectively. The contribution rate of NO3- in PM2.5 was the largest in winter, reaching 24.3%. In terms of spatial distribution, SNA contributed the most to PM2.5 in all regions, ranging from 32.8% to 39.7%, with the highest in northern Zhejiang and the lowest in southern Zhejiang. The SNA of all regions presented NO3->SO42->NH4+. Based on the backward trajectory clustering analysis, the main air sources of northern Zhejiang were the Yellow Sea-southern Jiangsu (autumn), northern Anhui (winter), East China Sea (spring), and western Jiangsu (summer) areas, with contribution rates of 38.11%, 35.28%, 37.46%, and 27.87%, respectively. The main air sources of western Zhejiang were the Yellow Sea-southern Jiangsu (autumn), southern Anhui (winter), eastern Zhejiang (spring), and northern Zhejiang (summer), with contribution rates of 38.11%, 37.50%, 46.55%, and 32.58%, respectively. The air of autumn, winter, spring, and summer in eastern Zhejiang were influenced by air masses from northern Hebei (36.07%), eastern Shandong (38.06%), East China Sea (30.17%), and southern Guangdong (34.43%), respectively. In autumn, winter, spring, and summer, southern Zhejiang was affected by air masses from the Yellow Sea (35.66%), northeast Anhui (34.44%), East China Sea (26.72%), and southern Fujian coast (35.00%), respectively. The regions in Zhejiang province showed large seasonal differences. The difference value between the maximum value of ρ(PM2.5) in the northwest and the lowest value in the southeast was 21.0 µg·m-3 and 20.5 µg·m-3 in autumn and winter, respectively; the difference values in spring and summer were 10.4 µg·m-3 and 6.1 µg·m-3. Thus, the northern air mass had a certain exogenous contribution to PM2.5 in autumn and winter in Zhejiang province. However, with the weakening of the northern air mass trajectory in spring and summer and the increasing contribution of the southern and east China Sea air mass to the air flow in Zhejiang province, PM2.5 pollution showed a trend of improvement.

Tailoring chiral optical properties by femtosecond laser direct writing in silica.

An object that possesses chirality, that is, having its mirror image not overlayed on itself by rotation and translation, can provide a different optical response to a left- or right-handed circular polarized light. Chiral nanostructures may exhibit polarization-selective optical properties that can be controlled for micro-to-nano optical element engineering. An attractive way to induce such complex nanostructures in three-dimension in glass is femtosecond laser direct writing. However, the mechanism of femtosecond laser induced chirality remains to be unveiled due to complex physical and chemical processes occurring during the ultrashort light-matter interaction. Here, a phenomenological model is proposed and is built on two-layers phase shifters to account for this laser-induced optical chirality in an initially achiral material (silica glass). This model is based on the observation that femtosecond laser induced nanogratings own two principal contributions to its aggregate birefringent response: a form and a stress-related one. By refining this formalism, a multilayer approach is developed to imprint on demand optical rotation. Values up to +/-60° at 550 nm within an optimal 80 µm thickness in silica glass are possible, corresponding to the highest value in a glass to date. These results provide new insights of circular-optical control in micro-nano optical manufacturing and open new opportunities for photonics applications.

Femtosecond laser direct writing multilayer chiral waveplates with minimal linear birefringence.

Chirality transfer from femtosecond laser direct writing in achiral transparent materials mainly originates from the interplay between anisotropic nanogratings and mechanical stress with non-parallel and non-perpendicular (oblique) neutral axes. Yet, the laser fabrication simultaneously induces non-negligible linear birefringence. For precise manipulation of circular polarization properties, as well as to unlock the full functionality, we report here a geometry-inspired multilayer method for direct writing of chiral waveplates with minimal linear birefringence. We perform a theoretical analysis of both circular and linear properties response for different multilayer configurations and achieve strong circular birefringence of up to -2.25 rad with an extinction ratio of circular birefringence to total linear birefringence of up to 5.5 dB at 550 nm. Our strategy enables the precise control of circular properties and provides a facile platform for chiral device exploration with almost no linear property existence.

Observation on the efficacy of sublingual immunotherapy with dust mite allergen for perennial allergic rhinitis and the mechanism of action on ILCs with ILC1s and ILC2s and ILC3s.

BACKGROUND: Allergic rhinitis (AR) is considered to be 1 of the most difficult diseases to treat globally. It has a serious impact on the quality of life and social economy of patients and has become an important global health problem. Several drugs have been recommended to treat AR, but their effectiveness and mechanism of action in these patients remain unclear. The purpose of this study will be to compare the efficacy and mechanism of action of 2 drugs for the treatment of AR (moderate to severe): a Dermatophagoides Farinae Drops Sublingual Immunotherapy and a Momethasone Furoate nasal spray as an adjunct to the treatment of subjects with AR. METHODS: A randomized, prospective, double-blind (patient and evaluator) clinical trial. The participants (n = 60) will be randomly distributed into 2 groups. The experimental group will receive a sublingual Immunotherapy for 3 months. The control group will receive the mometasone furoate nasal spray for 3 months. Before treatment, 1 month and 3 months after treatment, total nasal symptom score scale, Visual analogue Scale and Quality of Life questionnaire of rhinoconjunctivitis will be measured and Changes of the serums of IgE, interferon-γ, IL-4, IL-17, tumor necrosis factor-α, IL-5, IL-9, IL-13, IL-25, IL-33, vascular endothelial growth factor, TSLP and IL-22 in both groups. The measurements will be performed by the same researcher who was unaware of the participants' subgroup. DISCUSSION: We believe that the treatment of perennial AR with sublingual Immunotherapy and nasal hormones will be more effective in these patients. Furthermore, the sublingual Immunotherapy mainly acts mostly on the cellular immunity, while nasal hormones mainly act on local inflammatory responses. We expect to clarify which treatments are more effective and how they work in improving perennial AR.

RNA-methyltransferase Nsun5 controls the maternal-to-zygotic transition by regulating maternal mRNA stability.

BACKGROUND: RNA modification-induced ovarian dysgenesis appears to be necessary for ovary development. However, how m5 C (5-methylcytosine)-coordinating modificatory transcripts are dynamically regulated during oogenesis, and ovarian development is unknown. The purpose of this study was to determine whether NOP2/Sun RNA methyltransferase 5 (Nsun5) deletion leads to suppression of ovarian function and arrest of embryonic development. The regulation of mRNA decay and stability by m5 C modification is essential at multiple stages during the maternal-to-zygotic (MZT) transition. METHODS: Mouse ovaries and oocytes with Nsun5KO and the KGN cell line were subjected to m5 C identification, alternative splicing analysis and protein expression. BS-m5 C-seq, real-time polymerase chain reaction, Western blot, immunofluorescence and actinomycin D treatment assays were used. In particular, BS-m5 C-seq revealed a dynamic pattern of m5 C sites and genes in the ovaries between Nsun5KO and WT mice at the 2-month and 6-month stages. Diverse bioinformatic tools were employed to identify target genes for Nsun5. RESULTS: Here, a maternal mRNA stability study showed that deletion of the m5 C methyltransferase Nsun5 obstructs follicular development and ovarian function, which leads directly to inhibition of embryogenesis and embryo development. Dynamic analysis of m5 C revealed that the level of m5 C decreased in a time-dependent manner after Nsun5 knockout. Regarding the molecular mechanism, we found that Nsun5 deficiency caused a m5 C decline in the exon and 3'UTR regions that influenced the translation efficiency of Mitotic arrest deficient 2 like 2 (MAD2L2) and Growth differentiation factor 9 (GDF9) in the ovary. Mechanistic investigation of alternative splicing indicated that Nsun5KO triggers aberrant events in the exon region of Brd8. CONCLUSIONS: Nsun5 loss arrests follicular genesis and development in ovarian aging, indicating that Nsun5/m5 C-regulated maternal mRNA stabilization is essential for MZT transition.

Distributions and sources of traditional and emerging per- and polyfluoroalkyl substances among multiple environmental media in the Qiantang River watershed, China.

The presence of per- and polyfluoroalkyl substances (PFASs) in the food chain poses a threat to human health. Water and the atmosphere are the major transport pathways for PFASs in the environment, while water, soil and sediment are sinks. Herein, the concentrations and distributions of traditional and emerging PFASs in multi-environmental media samples in the Qiantang River watershed were comprehensively investigated. Twenty-five PFASs, including seven emerging PFASs, were identified. The concentrations in water, soil, sediment and PM2.5 ranged from 3.58 to 786 ng L-1, 0.72 to 12.3 ng g-1, 0.73 to 6.60 ng g-1, and 93.9 to 255 pg m-3, respectively, with mean concentrations of 149 ng L-1, 4.70 ng g-1, 4.31 ng g-1, and 156 pg m-3. Perfluorooctanoic acid (PFOA) was the dominant contaminant in water, soil, and sediment, and perfluoropentanoic acid (PFPeA) was the dominant contaminant in PM2.5. Hexafluoropropylene oxide-dimer acid (HFPO-DA) and 6 : 2 chlorinated polyfluorinated ether sulfonate (6 : 2 Cl-PFESA), as substitutes for PFOA and perfluorooctane sulfonate (PFOS), were detected, indicating the gradual replacement of traditional PFOA and PFOS in this area. Perfluoro-3,6-dioxaheptanoic acid (NFDHA), as a component of the aqueous film-forming foam FN-3, was first detected in this area. Short-chain PFASs were mainly distributed in water and PM2.5, while long-chain PFASs were distributed in the solid phase, such as soil, sediment, and PM2.5. Based on principal component analysis (PCA), the major PFAS sources were emulsifiers from fluorine polymerization and surface-active agents from the textile, papermaking, leather, and other industries. In addition, correlation analysis showed that water was the main source and transport pathway of short-chain perfluoroalkyl carboxylic acid (PFCA), HFPO-DA, and NFDHA in this area, while the atmosphere combined with PM2.5 was the main transport pathway for both short- and long-chain PFCAs, PFOS, and 6 : 2 Cl-PFESA.

Olfactory Ensheathing Cells Alleviate Facial Pain in Rats with Trigeminal Neuralgia by Inhibiting the Expression of P2X7 Receptor.

Trigeminal neuralgia (TN) is a common facial neuropathic pain that is mainly characterized by spontaneous or induced needling or electric shock pain in the innervation area of the trigeminal nerve. It is also referred to as "the cancer that never dies". The olfactory ensheathing cell (OEC) is a special glial cell in the nervous system that has a strong supportive function in nerve regeneration. Cell transplantation therapy is a useful treatment modality that we believe can be applied in TN management. In this study, OECs were transplanted into the ligation site of the infraorbital nerve of rats. We found that after the OEC transplantation, mechanical pain threshold in the face of the rats was significantly increased. Western blotting, immunofluorescence assay, and reverse transcription-quantitative polymerase chain reaction were performed on the trigeminal ganglia (TG) of model rats. The results revealed a decrease in the expression of P2X7 receptor (P2X7R) in the trigeminal ganglia. Our findings show that OEC transplantation has a good therapeutic effect on TN in rats, and that can reduce the expression of P2X7R in trigeminal ganglia. Therefore, we think that OEC transplantation may be a suitable treatment for TN.

The Role of m6A on Female Reproduction and Fertility: From Gonad Development to Ovarian Aging.

The growth and maturation of oocyte is accompanied by the accumulation of abundant RNAs and posttranscriptional regulation. N6-methyladenosine (m6A) is the most prevalent epigenetic modification in mRNA, and precisely regulates the RNA metabolism as well as gene expression in diverse physiological processes. Recent studies showed that m6A modification and regulators were essential for the process of ovarian development and its aberrant manifestation could result in ovarian aging. Moreover, the specific deficiency of m6A regulators caused oocyte maturation disorder and female infertility with defective meiotic initiation, subsequently the oocyte failed to undergo germinal vesicle breakdown and consequently lost the ability to resume meiosis by disrupting spindle organization as well as chromosome alignment. Accumulating evidence showed that dysregulated m6A modification contributed to ovarian diseases including polycystic ovarian syndrome (PCOS), primary ovarian insufficiency (POI), ovarian aging and other ovarian function disorders. However, the complex and subtle mechanism of m6A modification involved in female reproduction and fertility is still unknown. In this review, we have summarized the current findings of the RNA m6A modification and its regulators in ovarian life cycle and female ovarian diseases. And we also discussed the role and potential clinical application of the RNA m6A modification in promoting oocyte maturation and delaying the reproduction aging.

Integrity of cell-free DNA in maternal plasma extracellular vesicles as a potential biomarker for non-invasive prenatal testing.

OBJECTIVE: Large proportions of cell-free DNA (cfDNA) in plasma are localized in extracellular vesicles (EVs), which are secreted from placental cells. This study was conducted to reveal the integrity pattern of cfDNA in maternal plasma EVs (evcfDI) across gestation, and explore if evcfDI could be a potential biomarker in screening for aneuploid fetus in non-invasive prenatal testing (NIPT). METHODS: A total of 180 maternal plasma samples were collected during NIPT. Both evcfDNA and fetal evcfDNA (evcffDNA) were measured by quantitative PCR of LINE1 and SRY gene amplicons with different sizes. The evcfDI was calculated as the ratio of long to short fragments. RESULTS: evcfDI is not affected by gestational age; whereas evcffDI has a mild decreasing trend with increasing gestational age (P = 0.048). evcfDI is significantly and negatively correlated with maternal body mass index (BMI; calculated as weight in kilograms divided by the square of height in meters: ≤18.5, 18.5-25, and ≥25) (P < 0.01) and age (<35 and ≥35 years) (P < 0.01). Mean evcfDI decreases from 2.113 in euploid controls to 0.681 in those with an aneuploid fetus in NIPT (P = 0.003). CONCLUSION: Maternal clinical characteristics such as BMI and age could be innovative biomarkers to calibrate evcfDI, which was shown to be a potential indicator of an aneuploid fetus. Analysis of evcfDI based on quantitative PCR could serve as a novel, rapid, and low-cost NIPT strategy, which might facilitate testing at earlier gestations.

Vortex soliton oscillation in a mode-locked laser based on broadband long-period fiber grating.

Optical vortex beams (OVBs) have attracted much attention in diverse applications and spatiotemporal mode locking for optical soliton formation. In this Letter, a compact mode-locked (ML) vortex fiber laser is demonstrated based on a broadband long-period fiber grating near the dispersion turnaround point, where the group velocities between the core modes of ${{\rm LP}_{01}}$ and ${{\rm LP}_{11}}$ in a two-mode fiber are matched. The OVB pulses with first-order orbital angular momentum are oscillated through broadband mode conversion inside the cavity. The time-stretch dispersive Fourier transform method is also employed for the observation of vortex soliton buildup dynamics. The study of vortex soliton oscillation motivates the development towards controlling vortex modes in the ML fiber lasers.

Human placental mesenchymal stem cells ameliorate chemotherapy-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy.

BACKGROUND: The side effects of busulfan on male reproduction are serious, so fertility preservation in children undergoing busulfan treatment is a major worldwide concern. Human placental mesenchymal stem cells (hPMSCs) have advantages such as stable proliferation and lower immunogenicity that make them an ideal material for stimulating tissue repair, especially restoring spermatogenesis. The protective effects of hPMSCs in busulfan-induced Sertoli cells and in busulfan-treated mouse testes have not been determined. Our study aimed to elaborate the protective effect and potential mechanisms of hPMSCs in busulfan-treated testes and Sertoli cells. METHODS: First, we developed a mouse model of busulfan-induced testicular toxicity in vivo and a mouse Sertoli cell line treated with busulfan in vitro to assess the protective effect and mechanisms of hPMSC treatment on spermatogenesis. Then, the length, width, and weight of the testes were monitored using Vernier calipers. Furthermore, at 1 week and 4 weeks after the transplantation of hPMSCs, histological sections of testes were stained with hematoxylin-eosin, and the seminiferous tubules with fluid-filled cavities were counted. Through ELISA analysis, testosterone levels and MDA, SOD, LDH, and CAT activities, which are associated with ROS, were detected. Markers of ROS, proliferation (Ki67), and apoptosis (Annexin V) were evaluated by FACS. Next, the fluorescence intensity of proliferation markers (BrdU and SCP3), an antioxidant marker (SIRT1), a spermatogenesis marker (PLZF), and autophagy-related genes (P62 and LC3AB) were detected by fluorescence microscopy. The mRNA expression of γ-H2AX, BRCA1, PARP1, PCNA, Ki67, P62, and LC3 was determined by qRT-PCR. RESULTS: hPMSCs restored disrupted spermatogenesis, promoted improved semen parameters, and increased testosterone levels, testis size, and autophagy in the testis toxicity mouse model induced by busulfan. hPMSCs suppressed the apoptosis of Sertoli cells and enhanced their rate of proliferation in vitro. Additionally, hPMSCs protected against oxidative stress and decreased oxidative damage in the testis toxicity mouse model induced by busulfan. Furthermore, hPMSCs increased the expression of proliferation genes (PCNA and KI67) and decreased the mRNA levels of apoptotic genes such as γ-H2AX, BRCA1, and PARP1. CONCLUSIONS: This research showed that hPMSC injection ameliorated busulfan-induced damage in the testis by reducing apoptosis/oxidative stress and promoting autophagy. The present study offers an idea for a new method for clinical treatment of chemotherapy-induced spermatogenesis.

Dynamic vortex mode-switchable erbium-doped Brillouin laser pumped by high-order mode.

We experimentally demonstrated that the transversal vortex modes of an all-fiber erbium-doped Brillouin laser can be dynamically switched by using the high-order mode (HOM) of Brillouin pump (BP), which is used to achieve the oscillation of HOM inside the ring cavity. Core-mode conversion in a few-mode fiber (FMF) between the fundamental mode and HOM is obtained by cascading an acoustically induced fiber grating (AIFG) and a mode selection coupler (MSC) operating at the same wavelength region. Through frequency shift keying (FSK) modulation of the AIFG signal, the output transversal modes can be switched dynamically between LP01 and vortex modes, and the measured purities of output HOM are more than 82%. Moreover, the output Brillouin wavelength can also be tuned via altering the input wavelength of BP and the resonant response of AIFG. We have achieved HOM Brillouin-shifted laser output within the wavelength band from 1545-1560 nm. The output linewidth of the proposed Brillouin laser is less than 4 kHz.

Human amnion mesenchymal stem cells restore spermatogenesis in mice with busulfan-induced testis toxicity by inhibiting apoptosis and oxidative stress.

BACKGROUND: Before starting gonadotoxic therapies, cryopreservation of mature sperm has been proposed worldwide as a method for male fertility preservation and for enabling the conception of a healthy baby with assisted reproductive technology (ART); however, these technologies are not feasible for prepubertal boys and men with spermatogenic failure. Transplantation of mesenchymal stem cells has exhibited successful therapeutic benefits in restoring spermatogenesis via gonadal graft angiogenesis, transplanted cell clonogenesis, and disordered somatic compartment recovery. This study aimed to elucidate the fertility protective effects and the underlying mechanisms of human amnion mesenchymal stem cells (hAMSCs) against busulfan-induced testis toxicity. METHODS: An in vivo busulfan-induced testis toxicity mouse model and an in vitro busulfan-administered mouse Sertoli cell line were employed to evaluate the efficacy and mechanisms of hAMSC transplantation on male fertility preservation. The process of spermatogenesis was evaluated histologically, and the percentage of seminiferous tubules with vacuoles was evaluated by HE staining. Semen parameters were calculated by computer-assisted semen analysis. ELISA was employed to test the testosterone concentration and the levels of oxidative- and antioxidative-associated substances LDH, MDA, GR, SOD, GPx, and CAT. The rates of proliferation (Ki67), apoptosis (Annexin V), and ROS were measured by FACS. The fluorescence intensity of a marker of apoptosis (TUNEL) and a meiosis gene in spermatogenesis (SCP3) were detected by immunofluorescence assay. The expression of mRNA in germ cell-specific (GCS) genes (Dazl, Ddx4, and Miwi) and meiosis genes (Scp3, Cyclin A1, and Stra8) was tested by qPCR. The expression of antiapoptotic proteins (SURVIVIN and BCL2), apoptotic proteins (CASPASE3 and CASPASE9), GCS proteins (Dazl, Ddx4, and Miwi), and meiosis proteins (Scp3, Cyclin A1, and Stra8) was tested by western blotting. RESULTS: hAMSC transplantation following disruption by busulfan-induced testis toxicity restored spermatogenesis, elevating testosterone levels and enhancing testicular weight, size, and semen parameters in vivo. In addition, hAMSCs clearly ameliorated cell apoptosis, enhanced cell proliferation, repressed oxidative damage, and augmented oxidative defense in vivo and in vitro. Moreover, hAMSCs distinctly increased the expression of the GCS genes Dazl, Ddx4, and Miwi and the meiosis genes Scp3, Cyclin A1, and Stra8 in vivo. CONCLUSIONS: hAMSCs might represent a promising tool for the use in regenerative medicine, as these cells can restore spermatogenesis in a busulfan-induced testis toxicity mouse model and facilitate activity in a busulfan-administered mouse Sertoli cell line by resisting apoptosis and oxidative stress.

Exosomal miRNA-320a Is Released from hAMSCs and Regulates SIRT4 to Prevent Reactive Oxygen Species Generation in POI.

Human amniotic mesenchymal stem cells (hAMSCs) were previously shown to effectively rescue ovarian function in a premature ovarian insufficiency (POI) mouse model. The therapeutic mechanism of hAMSC-derived exosomes (hAMSC-Exos) is not fully understood. In this study, the therapeutic mechanism involved in exosomal microRNA-320a (miR-320a) and Sirtuin 4 (SIRT4) was investigated in POI mouse ovaries oocytes and human granulosa cells (hGCs) by fluorescence-activated cell sorting (FACS), hematoxylin and eosin (H&E) staining, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence experiments. hAMSC-Exos improved proliferation, inhibited apoptosis, and decreased the expression of SIRT4 and relative genes in POI hGCs and ovaries. hAMSC-Exos elevated ovarian function and prohibited SIRT4 expression in oogenesis. The therapeutic effects were attenuated when miR-320a was knocked down. hAMSC-Exos decreased the ROS levels in POI hGCs and oocytes and improved ovarian weight and litter size, except for the Exosanti-miR-320a/POI group. Finally, hAMSC-Exos reduced the SIRT4 and ROS levels in POI ovaries and hGCs. The downstream protein expression (ANT2, AMP-dependent kinase [AMPK], and L-OPA1) was downregulated in the hGCs-SIRT4KD group but disappeared in the Exosanti-miR-320a/POI group. Our study is the first to illustrate the therapeutic potential of hAMSC-Exos in POI. Exosomal miR-320 plays a key role in the hAMSC-Exos-mediated effects on ovarian function via SIRT4 signaling.

Exosomal miRNA-17-5p derived from human umbilical cord mesenchymal stem cells improves ovarian function in premature ovarian insufficiency by regulating SIRT7.

Premature ovarian insufficiency (POI) is clinically irreversible in women aged over 40 years. Although numerous studies have demonstrated satisfactory outcomes of mesenchymal stem cell therapy, the underlying therapeutic mechanism remains unclear. Exosomes were collected from the culture medium of human umbilical cord mesenchymal stem cells (hUMSCs) and assessed by electron microscopy and Western blot (WB) analysis. Then, exosomes were added to the culture medium of cyclophosphamide (CTX)-damaged human granulosa cells (hGCs), and the mixture was injected into the ovaries of CTX-induced POI model mice before detection of antiapoptotic and apoptotic gene expression. Next, the microRNA expression profiles of hUMSC-derived exosomes (hUMSC-Exos) were detected by small RNA sequencing. The ameliorative effect of exosomal microRNA-17-5P (miR-17-5P) was demonstrated by miR-17-5P knockdown before assessment of ovarian phenotype and function, reactive oxygen species (ROS) levels and SIRT7 expression. Finally, SIRT7 was inhibited or overexpressed by RNA interference or retrovirus transduction, and the protein expression of PARP1, γH2AX, and XRCC6 was analyzed. The ameliorative effect of hUMSC-Exos on POI was validated. Our results illustrated that hUMSC-Exos restored ovarian phenotype and function in a POI mouse model, promoted proliferation of CTX-damaged hGCs and ovarian cells, and alleviated ROS accumulation by delivering exosomal miR-17-5P and inhibiting SIRT7 expression. Moreover, our findings elucidated that miR-17-5P repressed PARP1, γH2AX, and XRCC6 by inhibiting SIRT7. Our findings suggest a critical role for exosomal miR-17-5P and its downstream target mRNA SIRT7 in hUMSC transplantation therapy. This study indicates the promise of exosome-based therapy for POI treatment.