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.

Laser ultrasonic improvement and its application in defect detection based on the composite coating method.

Herein, we studied the increasing tendency of photoacoustic (PA) conversion efficiency of the Au/polydimethylsiloxane (PDMS) composite. The thickness of the Au layer was optimized by modeling the PA process based on the Drude-Lorentz model and finite element analysis method, and corresponding results were verified. The results showed that the optimal Au thickness of the Au/PDMS composite was 35 nm. Finally, the Au/PDMS composites were coated onto the surface of aluminum alloys, which improved the thermoelastic laser ultrasonic (LU) signals to near 100 times. Besides, the defect mapping was performed by thermoelastic LU signals with Au/PDMS coating and ablation LU signals without coating; the Pearson correlation coefficient was higher than 0.95. The application in the defect detection in metal could provide guides for nondestructive detection on metals by laser ultrasound.

Long-term transplantation human menstrual blood mesenchymal stem cell loaded collagen scaffolds repair endometrium histological injury.

Menstrual blood mesenchymal stem cell (MBMSC) is a potential cell source for effective therapy for intrauterine adhesion (IUA). Collagen scaffold (CS) loaded with mesenchymal stem cells promotes endometrial regeneration in IUA model animals. However, role of combination of MBMSCs and CS in IUA therapy remains elusive. In particular, transplantation of MBMSCs over a long period of time requires more in-depth research. Here in this study, transplantation of human MBMSCs loaded on CS was applied for therapy for a long term rat IUA model. A rat IUA model characterized by lower number of endometrial glands and increased fibrosis was established. At 90 days after transplantation of the human MBMSC-loaded CS, expression of HuNu, a human protein, was identified in the uteri of the transplanted IUA model rats. The transplantation increased the number of endometrial glands and decreased the fibrotic areas significantly. Moreover, transplantation of the human MBMSC-loaded CS decreased the Collagen I and increased the CK 18 significantly. Immunoblotting assay results further proved the downregulation of Collagen I and the upregulation of CK 18. Together, endometrium regeneration promoted by human MBMSC-loaded CS was demonstrated in a long term rat model of IUA, shedding a new light on the role of human MBMSCs in the therapy for IUA.

Stimulation of both inspiratory and expiratory muscles versus diaphragm-only paradigm for rehabilitation in severe chronic obstructive pulmonary disease patients: a randomized controlled pilot study.

BACKGROUND: Chronic obstructive pulmonary disease (COPD), a progressive lung disease, might improve with neuromuscular electrical stimulation. No trials on inspiratory plus expiratory neuromuscular electrical stimulation have been conducted yet. AIM: The aim of this study was to evaluate the safety and effectiveness of inspiratory plus expiratory neuromuscular electrical stimulation in subjects with severe COPD. DESIGN: This was a multicenter, prospective, randomized controlled trial. SETTING: The subjects were outpatients enrolled from Beijing Chao-Yang Hospital affiliated with Capital Medical University, Tianjin Chest Hospital, and the First Hospital of Hebei Medical University. POPULATION: Subjects had stable COPD with severe respiratory impairment. METHODS: Using a computer statistical software, 120 stable subjects were randomly allocated (1:1) to receive inspiratory plus expiratory neuromuscular electrical stimulation (study group) and diaphragm pacing (control group). Demographic and clinical data were collected before, and after 2, and 4 weeks of the trial. The intention-to-treat analysis was conducted. The primary outcome was to analyze the changes in functional exercise capacity, estimated as six-minute walk distance (6MWD), following electrical stimulation for 4 weeks. The secondary outcomes were changes in modified Medical Research Council score, forced expiratory volume in 1 second (FEV1), FEV1% predicted, and FEV1 ratio forced vital capacity (FEV1/FVC) following electrical stimulation for 4 weeks. RESULTS: The change in 6MWD was greater in the study group (65.53±39.45 m) than in the control group (26.66±32.65 m). The mean between-group difference at the fourth week was 29.07 m (95% confidence interval, 16.098-42.035; P<0.001). There were no significant between-group differences in the secondary outcomes after 4 weeks of electrical stimulation. For GOLD-4 COPD subjects, FEV1 and FEV1/FVC improved in the study group (P<0.05). No electrical stimulation-related serious adverse events were observed in either group. CONCLUSIONS: 6MWD were increased significantly, without adverse events, after four weeks of treatment of inspiratory plus expiratory neuromuscular electrical stimulation in stable patients with severe COPD, suggesting that this protocol benefits COPD rehabilitation. CLINICAL REHABILITATION IMPACT: The results of this study suggest that the simultaneous use of inspiratory plus expiratory neuromuscular electrical stimulation as an adjunct therapy may improve the functional exercise capacity of severe stable COPD subjects.

Age-related changes in expression of lysine48 and lysine63 ubiquitin linkages in dopamine neurons of midbrain in mice.

Ubiquitination of target proteins is mediated via different ubiquitin lysine (K) linkages and determines the protein fates. In particular, K48 ubiquitin linkage targets proteins for degradation, whereas K63 ubiquitin linkage plays a nondegradative role. Parkinson's disease is an age-onset neurodegenerative disorder, which shows selective loss of dopamine neurons in substantia nigra pars compacta (SNC) and ubiquitinated protein aggregates. However, age-related expression of K48 and K63 ubiquitin linkages in SNC dopamine neurons remains elusive. We thus sought to explore the expression of K48 and K63 ubiquitin linkages in dopamine neurons in SNCs of mice at different ages with morphological and biochemical assays. Here our results indicated that in 5-week-old mice, dopamine neurons presented higher levels of K48 and K63 ubiquitin linkages than nondopamine neural cells. Aging promoted the formation of protein aggregates that are positive for both K48 and K63 ubiquitin linkages, together with tyrosine hydroxylase, a dopamine neuron marker. Moreover, 21-month-old mice showed fewer neural cells and tyrosine hydroxylase positive neurons in the SNCs than younger mice. Through biochemical analysis, the 21-month-old mice were shown to express more K48 ubiquitin linkages and less tyrosine hydroxylase and NeuN than the 5-week-old mice. These results suggest the first time that expression of K48 and K63 ubiquitin lysine linkages in midbrain dopamine neurons is age-related and may be involved in the loss of dopamine neurons.

Lockdown effects on total suspended solids concentrations in the Lower Min River (China) during COVID-19 using time-series remote sensing images.

The COVID-19 pandemic in China in the winter-spring of 2019-2020 has decreased and even stopped many human activities. This study investigates whether there were any changes in the water quality of the Lower Min River (China) during the lockdown period. The time-series remote sensing images from November 2019 to April 2020 was used to examine the dynamics of the river's total suspended solids (TSS) concentrations in the period. A new remote sensing-based prototype was developed to recalibrate an existing algorithm for retrieving TSS concentrations in the river. The Nechad and the Novoa algorithms were used to validate the recalibrated algorithm. The results show that the recalibrated algorithm is highly consistent with the two algorithms. All of the three algorithms indicate significant fluctuation in TSS concentrations in the Lower Min River during the study period. February (COVID-19 lockdown period) has witnessed a 48% fall in TSS concentration. The TSS in March-April showed a progressive and recovery back to normal levels of pre-COVID-19. The spatiotemporal change of TSS has worked as a good indicator of human activities, which revealed that the decline of TSS in the lockdown period was due largely to the substantially-reduced discharges from industrial estates, densely-populated city center, and river's shipping. Remote sensing monitoring of the spatiotemporal changes of TSS helps understand important contributors to the water-quality changes in the river and the impacts of anthropogenic activities on river systems.

Investigation of the mechanism and influence of laser wavelength and energy on laser opto-ultrasonic dual detection.

Laser opto-ultrasonic dual (LOUD) detection, which uses laser irradiation of samples to generate spectral and ultrasonic signals simultaneously, can perform multimodal detection of element composition and structural property. As such, it has been applied to the detection of additive manufacturing (AM) components. Further, optimized parameters lead to better detection results. To the best of our knowledge, however, there is no study on the effect of laser properties on LOUD detection. Therefore, we studied the mechanism and influence of laser wavelength and energy on LOUD detection. In this work, the intensity, signal-to-noise ratio (SNR), and stability evolution of the laser excitation spectrum and ultrasonic signals at different wavelengths and energies were analyzed. It was found in the plasma evolution that high electron number density means a large amount of ablated mass generated, which was favorable for laser ultrasonic excitation and can produce higher SNR and a more stable signal. However, it also led to more atoms of the ground-state, which resulted in the self-absorption effect and reduced spectrum intensity in the spectrum analysis. Therefore, with self-absorption correction, better stability, and higher signal intensity, an SNR of spectral and ultrasonic signals can be obtained using 355 nm laser excitation at optimal energy. As a result, in the quantitative analysis of Cu and Si elements by LOUD detection, the determination coefficients (R2) were higher than 0.995, and the average relative errors were less than 2.5%, the limit of detection could reach the order of 100 ppm. Further, the defect size of 0.55 mm in the wire +arc additive manufacturing sample was detected by LOUD detection, and the average relative error was 5.59% compared with the digital radiography results, which indicate that laser wavelength and laser energy affect the intensity and stability of spectral and ultrasonic signals in LOUD detection, which means selecting appropriate laser parameters is important to obtain a high precision detection.

Simultaneous Compositional and Grain Size Measurements Using Laser Opto-Ultrasonic Dual Detection for Additive Manufacturing.

Metal-based additive manufacturing (AM) is a disruptive technique with great potential across multiple industries; however, its manufacturing quality is unstable, leading to an urgent requirement for component properties detection. The distribution of grain size has an important effect on many mechanical properties in AM, while the distribution of added elements, such as titanium (Ti), has a measurable effect on the grain size of an aluminum (Al) alloy. Therefore, the detection of the distributions of grain size and elements is of great significance for AM. In this study, we investigated the distribution of grain size and elements simultaneously for wire + arc additive manufacturing (WAAM) with an Al alloy using laser opto-ultrasonic dual (LOUD) detection. The average grain size obtained from the acoustic attenuation of ultrasonic signals was consistent with the results of electron backscatter diffraction (EBSD), with a coefficient of determination (R2) of 0.981 for linear fitting. The Ti element distribution obtained from optical spectra showed that the enrichment of Ti corresponded to the grain refinement area in the detected area. The X-ray diffraction (XRD) spectra showed that the spectral peaks were moved from Al to AlTi and Al2Ti forms in the Ti-rich areas, which confirmed the LOUD results. The results indicated that LOUD detection holds promise for becoming an effective method of analyzing the mechanical and chemical properties of components simultaneously, which could help explain the complex physical and chemical changes in AM and ultimately improve the manufacturing quality.

A fluorescent sensor for folic acid based on crown ether-bridged bis-tetraphenylethylene.

Aggregation-induced emission (AIE) provides a new strategy for preparing fluorescent sensors in aggregated state. In this paper, a series of crown ether-bridged bis-tetraphenylethylene compounds were synthesized in 78-84% yield by a simple procedure. The molecules exhibited excellent AIE properties in THF/H2O solutions and solid films. The investigation on sensing abilities for various biomolecules and metal ions suggested that Bis-TPE-1 possessed obvious response to folic acid, with fluorescence enhancement and blue shift of maximum emission wavelength from 380 nm to 365 nm. The detection limit for folic acid was 6.36 × 10-7 M, and the sensor's selectivity for folic acid was little interfered by the other species. The sensor mechanism was studied by FT-IR, 1H NMR, MS spectra and fluorescence Jobs' plot. The selective sensor for folic acid was applied in test paper and the analyses of real samples of mung bean and spinach. The superior bioimaging performance of Bis-TPE-1 for sensing folic acid was confirmed by the live cell imaging experiments, which indicated its good practical application potential for detecting folic acid.

[A case of the nasal septum pyogenic granuloma and literature].

A 56-year-old male had presented with left nasal bleeding repeatedly for 4 days. The pathological examination after resection showed pyogenic granuloma. Its etiology and pathogenesis, clinical features, pathological features and treatments were reviewed.

[The aspergillus laryngotracheobronchitis. A case report and literature review].

The patient presented with dry cough, lending, fever and progressive dyspnea for two weeks. The patient had a prior respiratory infection history and the symptoms were not obvious, Early X-ray showed lung infection. Under the fibrolaryngoscope, the lingual surfaces of the epiglottis, epiglottic vallecula, and bilateral vocal cords were covered by yellow pseudomembrane. The motion of vocal cords was normal with poor glottic closure, and no ulcer was noted. Endotracheal mucosa was swelling and congested with an uneven surface, and purulent discharge and pseudomembrane was formed. Pathological examination revealed Aspergillus. The disease was diagnosed as Aspergillus laryngotracheobronchitis.

Upregulation of GRP78 and GRP94 and its function in chemotherapy resistance to VP-16 in human lung cancer cell line SK-MES-1.

The upregulation of GRP78 and GRP94 under the induction of A23187 and its function in drug resistance to etoposide (VP-16) was investigated in human lung cancer cell line SK-MES-1. The expression of GRP78 and GRP94 induced by A23187 at different concentrations was analyzed by RT-PCR and Western blotting. Cell survival to VP-16 was determined using a colony-formation assay. The expression of GRP78 and GRP94 in the cells was found to correspond well with the cell survival to VP-16. The results showed that upregulation of GRP78 and GRP94 can significantly confer the chemoresistance to VP-16 in human lung cancer cell line SK-MES-1.

VP-16 resistance in the NCI-H460 human lung cancer cell line is significantly associated with glucose-regulated protein78 (GRP78) induction.

AIM: To investigate the relationship between the expression of glucose-regulated protein (GRP78) and resistance to VP-16 in the NCI-H460 cell line. METHODS: RT-PCR, real-time RT-PCR and Western blotting were used in analyzing the expression of GRP78 at mRNA and protein levels in the NCI-H460 cell line induced by A23187 at different concentrations. Cell survival with VP-16 was determined using a colony-formation assay with the account of IC50. RESULTS: The expression of GRP78 at both the mRNA and protein levels was higher in the NCI-H460 cell line induced by A23187. A23187 treatment resulted in up to 4.8-fold elevation of GRP78 mRNA and up to 3.2-fold elevation of GRP78 protein in the experimental cells compared to the controls, all in a dose-dependent manner. The IC50s for VP-16 in the cells pretreated with different concentrations of A23187 (0, 1, 2, 4 and 6 microM) were: 12.11 +/- 0.83, 12.68 +/- 1.04, 25.82 +/- 1.83, 37.46 +/- 1.89 and 45.19 +/- 2.34 microM, respectively. Compared to the control, there was a significant elevation of IC50 for VP-16 in the cells pretreated with A23187. Survival curve analysis also showed that the induction of A23187 caused a significantly longer survival for the cells subjected to VP-16 treatment (p < 0.05). CONCLUSION: A23187 treatment is highly effective for the induction of GRP78 and subsequent development of resistance to VP-16 in the human lung cancer NCI-H460 cell line. Based on the trend of the change in IC50 and the expression of GRP78 in differently exposed cells, we conclude that the induction of GRP78 by A23187 is significantly associated with the resistance to VP-16.