METTL3-mediated m6A methylation of DNMT1 promotes the progression of non-small cell lung cancer by regulating the DNA methylation of FOXO3a.

Background: The aim of this study was to investigate the effect of DNA methylation of Fork Head Box O3 (FOXO3a) on the process of epithelial-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC). Methods: The expressions of FOXO3a, DNA methyltransferase 1 (DNMT1), METTL3, and EMT-related proteins (E-cadherin and N-cadherin) were measured. The influence of 5-Aza-dC and DNMT1 on the methylation level in the promoter region of FOXO3a was examined through the application of methylation-specific PCR (MSP). Chromatin immunoprecipitation (ChIP) was employed to detect binding between DNMT1 and the FOXO3a promoter. Methylated RNA immunoprecipitation (MeRIP) was utilized to evaluate the level of DNMT1 N6-methyladenosine (m6A) methylation. The assessment of cell viability and invasion abilities of A549 cells was performed using Cell Counting Kit-8 (CCK-8) and Transwell assays, respectively. NSCLC xenograft mouse models were established by subcutaneously injected treated A549 cells into nude mice. Results: The expression levels of DNMT1 and DNA methylation level FOXO3a were found to be significantly increased, whereas FOXO3a expression was considerably decreased in NSCLC cell lines and NSCLC tumor tissues. Both 5-Aza-dC treatment and DNMT1 knockdown resulted in the down-regulation of DNA methylation levels of FOXO3a while simultaneously up-regulating the expression of FOXO3a. A ChIP assay demonstrated that DNMT1 has the ability to bind to the promoter region of FOXO3a. Furthermore, the knockdown of DNMT1 promoted E-cadherin expression, but inhibited expression of N-cadherin, cell viability, and invasion ability. However, the knockdown of FOXO3a hindered the effect of DNMT1 knockdown on EMT, cell viability, and invasion ability of A549 cells. This was evidenced by decreased E-cadherin expression and increased N-cadherin expression, as well as increased cell viability and invasion ability. Increased expression of DNMT1 resulted from m6A methylation of DNMT1, which was mediated by METTL3. Overexpression of DNMT1 decreased of E-cadherin expression while increased N-cadherin expression, cell viability, and invasion ability in METTL3-shRNA treated A549 cells. In xenograft mouse models, DNMT1 knockdown significantly reduced tumor volumes and tumor weight. DNMT1 knockdown upregulated the expression of FOXO3a and E-cadherin, while downregulated N-cadherin expression in vivo. Conclusion: METTL3-mediated m6A methylation of DNMT1 up-regulates FOXO3a promoter methylation, thereby promoting the progression of NSCLC.

Non-operative treatment strategy for appendiceal abscess in children under 3 years old: a retrospective observational study.

Background: There are few studies on appendiceal abscess with appendicolith in children under 3 years old. This study aims to explore the success rate of non-surgical treatment of appendiceal abscess and assess the potential influence of an appendicolith on non-surgical treatment outcomes in children under 3 years old. Methods: The clinical data of children under 3 years old who were diagnosed with appendiceal abscess at the Wuhan Children's Hospital, China, from February 2013 to May 2020 were collected. According to the findings of ultrasonography and CT imaging, they were divided into two groups, namely, the appendicolith group and the non-appendicolith group. Results: A total of 94 children with appendiceal abscess were identified, meeting the specified study criteria, and categorized into two groups, namely, the appendicolith group (n = 51, 54.3%) and the non-appendicolith group (n = 43, 45.7%). Non-surgical treatment was unsuccessful in six out of the 94 children, yielding an overall success rate of 93.6% for non-surgical management of appendiceal abscess in children under 3 years old. The success rate for non-surgical treatment in the appendicolith subgroup was 90.2%, whereas that for the non-appendicolith subgroup was 97.7%. No statistically significant distinction was observed between the two groups (P = 0.292). Likewise, there were no significant differences in gender, age, duration of symptoms, fever, vomiting, diarrhea, rebound pain, white blood cell count, C-reactive protein level, and abscess cross-sectional area between the appendicolith group and the non-appendicolith group. However, there is a statistical difference in tenderness in the right lower abdomen. Conclusion: Non-surgical treatment of appendiceal abscess has a high success rate and can be considered an effective treatment strategy. In pediatric patients under 3 years old without evidence of complete intestinal obstruction or diffuse peritonitis, non-surgical treatment may be considered for appendiceal abscess.

Electric-Field-Driven Printed 3D Highly Ordered Microstructure with Cell Feature Size Promotes the Maturation of Engineered Cardiac Tissues.

Engineered cardiac tissues (ECTs) derived from human induced pluripotent stem cells (hiPSCs) are viable alternatives for cardiac repair, patient-specific disease modeling, and drug discovery. However, the immature state of ECTs limits their clinical utility. The microenvironment fabricated using 3D scaffolds can affect cell fate, and is crucial for the maturation of ECTs. Herein, the authors demonstrate an electric-field-driven (EFD) printed 3D highly ordered microstructure with cell feature size to promote the maturation of ECTs. The simulation and experimental results demonstrate that the EFD jet microscale 3D printing overcomes the jet repulsion without any prior requirements for both conductive and insulating substrates. Furthermore, the 3D highly ordered microstructures with a fiber diameter of 10-20 µm and spacing of 60-80 µm have been fabricated by maintaining a vertical jet, achieving the largest ratio of fiber diameter/spacing of 0.29. The hiPSCs-derived cardiomyocytes formed ordered ECTs with their sarcomere growth along the fiber and developed synchronous functional ECTs inside the 3D-printed scaffold with matured calcium handling compared to the 2D coverslip. Therefore, the EFD jet 3D microscale printing process facilitates the fabrication of scaffolds providing a suitable microenvironment to promote the maturation of ECTs, thereby showing great potential for cardiac tissue engineering.

Clinical Efficacy of Thoracoscopic Surgery with the da Vinci Surgical System versus Video-Assisted Thoracoscopic Surgery for Lung Cancer.

Objective: To assess the clinical efficacy of thoracoscopic surgery with the da Vinci surgical system versus video-assisted thoracoscopic surgery (VATS) for lung cancer. Methods: From August 2019 to December 2020, 193 patients with lung cancer assessed for eligibility scheduled for surgery in our hospital were recruited and assigned at a ratio of 1 : 1 to receive VATS (control group) or thoracoscopic surgery with the da Vinci surgical system (research group). The primary measurement is the clinical efficacy of the two surgical modalities. Results: The baseline features of the research group were comparable with those of the control group (P > 0.05). Besides, the two groups showed similar tumor types, tumor locations, and clinicopathological staging (P > 0.05). Da Vinci surgical system-assisted thoracoscopic surgery had short operative time, less intraoperative blood loss, better lymph node dissection, and lower intraoperative conversion rates compared to VATS. Compared with the control group, the research group had significantly higher postoperative forced expiratory volume in one second (FEV1), forced vital capacity (FVC), maximal voluntary ventilation (MVV), the functional assessment of cancer therapy-general module (FACT-G) of the FACT-lung (FACT-L) Chinese version V4.0, lung cancer-specific module scores, and total scores (P < 0.05). The research group showed better postoperative drainage volume, shorter intubation duration, and length of hospital stay and a lower incidence of complications versus the control group (P < 0.05). The da Vinci surgical system reduced the probability of intraoperative mistakes and better ensured a safe and satisfactory surgery. Conclusion: The thoracoscopic surgery with the da Vinci surgical system better reduces intraoperative and postoperative bleeding, shortens drainage and intubation duration, enhances the lung function and survival quality of patients, and lowers the risk of surgical mistakes to ensure surgical safety versus VATS.

A Large-Scale Screening Responding Sporadic Epidemic of COVID-19 in China by an Integrated Health-Care System.

Introduction: In the post-pandemic period of COVID-19, the majority of cities in China try to balance the normalization of epidemic prevention and social-economic development. However, the appearance of asymptomatic infected patients poses threats to public health, which might be infectious without clinical symptoms and only be detected by testing approaches. Methods: Along with the appearance of one symptomatic case, a regional large-scale screening program was carried out in Shenzhen City charged by a regionally integrated healthcare system. After describing the screening program, a retrospective cross-sectional study for the screening outcome and efficacy was conducted. Discussion: According to the screening results, the asymptomatic case was infectious and their close contacts should be quarantined cautiously as the close contacts of symptomatic cases. Besides, after integrating medical resources in Luohu district of Shenzhen, the medical capability of Luohu district improved greatly which could be demonstrated in inspection and organization abilities in this screening program. Conclusion: The large-scale screening contributed to preventing epidemic transmission. In the post-pandemic period, regular surveillance of asymptomatic cases and rapid response capability for emergent screening program are both crucial for the prevention and control of COVID-19 epidemic. The integrated healthcare system coordinating regional medical institutions and optimizing regional medical recourse has advantages to address public health emergencies.

Neoadjuvant Nivolumab and Chemotherapy in Patients with Locally Advanced Non-Small Cell Lung Cancer: A Retrospective Study.

BACKGROUND: Patients with locally advanced (stage III) non-small cell lung cancer (NSCLC) demonstrate broad anatomic heterogeneity with modest survival benefits. Immune checkpoint inhibitors (ICIs) have shown survival benefit in metastatic NSCLC. We conducted this study to evaluate the efficacy and safety of neoadjuvant nivolumab in combination with chemotherapy in the treatment of this population. METHODS: We retrospectively evaluated patients with locally advanced NSCLC receiving neoadjuvant nivolumab and chemotherapy (paclitaxel with carboplatin) followed by surgery at our institution from January 2019 to January 2020. RESULTS: A total of 46 eligible patients, 26 males, and 20 females were diagnosed with NSCLC in a stage IIIA (30 cases) and IIIB (16 cases) to receive neoadjuvant treatment. The treatment was well tolerated with just 7 (15.2%) typical immune-related adverse events (hyperthyroidism, hyperglycemia, and rash) recorded. A total of 45 patients underwent surgical resection, and 43 (95.6%) of them achieved a R0 resection. No major surgical complications were observed. There was a complete response (CR), partial response (PR), stable disease (SD), and progressive disease (PD) in 2 (4.3%), 26 (56.5%), 17 (37.0%), 6 (26.1%), and 1 (2.2%) patients. Eight patients resulted in a major pathological response (MPR) (17.4%) and 24 patients had a pathological complete response (pCR) (52.2%). At the time of data cutoff (June 1, 2021), the median follow-up period was 15.5 months (IQR 3.9-29) and 27 (60%) of 45 patients who had tumor resection were progression free. At 24 months, progression-free survival was 45.8% and overall survival was 79.9%. CONCLUSION: Nivolumab plus paclitaxel and carboplatin could be a potential neoadjuvant regimen for patients with locally advanced NSCLC, rendering a potentially lethal disease to one that is curable.

The Electric-Field-Driven Fusion Jetting 3D Printing for Fabricating High Resolution Polylactic Acid/Multi-Walled Carbon Nanotube Composite Micro-Scale Structures.

Existing 3D printing techniques are still facing the challenge of low resolution for fabricating polymer matrix composites, inhibiting the wide engineering applications for the biomedical engineering (biomimetic scaffolds), micro fuel cells, and micro-electronics. In order to achieve high resolution fabrication of polylactic acid (PLA)/multi-walled carbon nanotube (MWCNT) composites, this paper presents an electric-field-driven (EFD) fusion jetting 3D printing method by combining the mixing effect and material feeding of the micro-screw and the necking effect of Taylor cone by the EFD. The effects of main process parameters (the carbon loading, the voltage, the screw speed, and the printing speed) on the line width and the printing quality were studied and optimized. To demonstrate the printing capability of this proposed method, meshes with line width of 30 µm to 100 µm and 1 wt.% to 5 wt.% MWCNT for the application of conductive biomimetic scaffold and the anisotropic flexible meshes were prepared. The electrical properties were investigated to present the frequency dependence of the alternating current conductivity and the dielectric loss (tanδ), and the microstructures of printed structures demonstrated the uniformly dispersed MWCNT in PLA matrix. Therefore, it provides a new solution to fabricate micro-scale structures of composite materials, especially the 3D conductive biomimetic scaffolds.

Synthesis of Active Hexafluoroisopropyl Benzoates via a Multicomponent Reaction.

Herein, we describe an efficient, practical free-metal rapid access to active hexafluoroisopropyl benzoates from anthranils, hexafluoroisopropanol, and N-alkoxy α-halogenoacetamides. Notably, this process includes anthranils that underwent a distinct pattern reaction. The protocol has good functional group tolerance and a broad substrate scope. Using a simple and general method, we accomplished potential synthetic application of active ester.

Integrative transcriptomics and proteomics analysis constructs a new molecular model for ovule abortion in the female-sterile line of Pinus tabuliformis Carr.

Ovule development is critical to plant reproduction and free nuclear mitosis of megagametophyte (FNMM) is vital for ovule development. However, most results of ovule development were based on the studies in angiosperms, and its molecular regulation remained largely unknown in gymnosperms, particularly, during FNMM. In this context, we studied the genome-wide difference between sterile line (SL) and fertile line (FL) ovules using transcriptomics and proteomics approaches in Pinus tabuliformis Carr. Comparative analyses revealed that genes involved in DNA replication, DNA damage repair, Cell cycle, Apoptosis and Energy metabolism were highlighted. Further results showed the low expressions of MCM 2-7, RRM1, etc. perhaps led to abnormal DNA replication and damage repair, and the significantly different expressions of PARP2, CCs1, CCs3, etc. implied that the accumulated DNA double-stranded breaks were failed to be repaired and the cell cycle was arrested at G2/M in SL ovules, potentially resulting in the occurrence of apoptosis. Moreover, the deficiency of ETF-QO might hinder FNMM. Consequently, FNMM stopped and ovule aborted in SL ovules. Our results suggested a selective regulatory mechanism led to FNMM half-stop and ovule abortion in P. tabuliformis and these insights could be exploited to investigate the molecular regulations of ovule development in woody gymnosperms.

Ce-Based Nanoparticles Loaded with Cisplatin for Tumour Radiotherapy.

The combination of radiotherapy and chemotherapy is a common and useful treatment mode for tumours. But traditional methods inevitably lead to a variety of side effects. A drug delivery system (DDS), which has good biocompatibility and strong anti-tumour ability, is expected to solve this problem. Studies have shown that Ce-based nanoparticles (NPs) have good radiosensitization effect through the photoelectric effect. Hence, cisplatin-loaded LiLuF4 :Ce3+scintillation NPs (NP + Cis) were first constructed in this study, which was synthesized by the crystal precipitation method and characterized by transmission electron microscopy (TEM). Subsequently, its toxicity was verified, and the radiosensitization effect and basic radiosensitization mechanism on tumour cells and tumour-bearing mice were researched. Results showed that NP + Cis triggered massive DNA damage and effectively inhibited cell viability in vitro under the exposure of X-ray irradiation (IR). Moreover, the experiments in vivo showed that the NP + Cis had higher biosafety, which could absorb enough irradiation and produce a synergistic inhibitory effect on tumours through the releasing of Cis. NP + Cis can improve the performance of DDS in chemoradiotherapy.

Chemical constituents from common vetch (Vicia sativa L.) and their antioxidant and cytotoxic activities.

Vicia sativa L. (common vetch) is a potential food source for both human beings and animals because of its abundant nutritional composition. There is a lack of phytochemical study on the whole plant, and thus the objective of this study was to investigate the isolation of phytochemicals and evaluate their biological activities. A new flavanol, (2R,3S)-3,3'-dihydroxy-4',7-dimethoxyflavanol (1), together with nine known compounds, two flavones (2-3), one coumarin (4), and six oleanane triterpenoids (5-10), was obtained from Vicia sativa L.. The structure of the new compound 1 was determined via its NMR spectra, IR and CD data. Compound 3 displayed the potential of the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging effect in antioxidant test. In terms of cytotoxic activities, compound 3 showed moderate cytotoxic activities against three human tumor cells, especially HeLa cells.

An efficient synthesis of oxazolines via a cascade reaction between azaoxyallyl cations and 1,2-benzisoxazoles.

A formal [3 + 2] cycloaddition reaction between the C and O terminals of azaoxyallyl cations formed in situ and 1,2-benzisoxazoles has been realized. This one-pot cycloaddition method provided an effective and practical pathway to synthesize oxazoline in good yields under mild conditions. The title products exhibited unique fluorescence properties.

Penile pyoderma gangrenosum: Successful treatment with thalidomide.

Pyoderma gangrenosum (PG) is a rare ulcerating inflammatory neutrophilic dermatosis. Different clinical manifestations have been described, including ulcerative, pustular and bullous, and vegetative variants. Classic PG usually occurs on the lower extremities (~70% of cases) but can also involve the hands, head, neck, and scrotum. Genital involvement of PG has rarely been reported. Treatment of the genital PG is usually difficult and resistance to conventional therapeutic regimens was frequently observed. The present authors reported a 16-year-old male patient who presented with progressive genital ulceration for 3 weeks. He was treated successfully low dose thalidomide (50 mg/d) and minocycline.

Uniparental chicken offsprings derived from oogenesis of chicken primordial germ cells (ZZ).

Cloning (somatic cell nuclear transfer) in avian species has proven unachievable due to the physical structure of the avian oocyte. Here, the sexual differentiation of primordial germ cells with genetic sex ZZ (ZZ PGCs) was investigated in female germline chimeric chicken hosts with the aim to produce uniparental offspring. ZZ PGCs were expanded in culture and transplanted into the same and opposite sex chicken embryos which were partially sterilized using irradiation. All tested chimeric roosters (ZZ/ZZ) showed germline transmission with transmission rates of 3.2%-91.4%. Unexpectedly, functional oogenesis of chicken ZZ PGCs was found in three chimeric hens, resulting in a transmission rate of 2.3%-27.8%. Matings were conducted between the germline chimeras (ZZ/ZZ and ZZ/ZW) which derived from the same ZZ PGCs line. Paternal uniparental chicken offspring were obtained with a transmission rate up to 28.4% and as expected, all uniparental offspring were phenotypic male (ZZ). Genotype analysis of uniparental offsprings was performed using 13 microsatellite markers. The genotype profile showed that uniparental offspring were 100% genetically identical to the donor ZZ PGC line, shared 69.2%-88.5% identity with the donor bird. Homozygosity of the tested birds varied from 61.5% to 84.6%, which was higher than the donor bird (38.5%). These results demonstrate that male avian ZZ PGCs can differentiate into functional ova in an ovary, and uniparental avian clones are possible. This technology suggests novel approaches for generating genetically similar flocks of birds and for the conservation of avian genetic resources.

Safety and efficacy of low fluence, high repetition rate versus high fluence, low repetition rate 810-nm diode laser for axillary hair removal in Chinese women.

BACKGROUND: High-fluence diode lasers with contact cooling have emerged as the gold standard to remove unwanted hair. Lowering the energy should result in less pain and could theoretically affect the efficacy of the therapy. OBJECTIVE: To compare the safety and efficacy of a low fluence high repetition rate 810-nm diode laser to those of a high fluence, low repetition rate diode laser for permanent axillary hair removal in Chinese women. METHODS: Ninety-two Chinese women received four axillae laser hair removal treatments at 4-week intervals using the low fluence, high repetition rate 810-nm diode laser in super hair removal (SHR) mode on one side and the high fluence, low repetition rate diode laser in hair removal (HR) mode on the other side. Hair counts were done at each follow-up visit and 6-month follow-up after the final laser treatment using a "Hi Quality Hair Analysis Program System"; the immediate pain score after each treatment session was recorded by a visual analog scale. RESULTS: The overall median reduction of hair was 90.2% with the 810-nm diode laser in SHR mode and 87% with the same laser in HR mode at 6-month follow-up. The median pain scores in SHR mode and in HR mode were 2.75 and 6.75, respectively. CONCLUSION: Low fluence, high repetition rate diode laser can efficiently remove unwanted hair but also significantly improve tolerability and reduce adverse events during the course of treatment.

Several insights into the preprocessing of electrograms in atrial fibrillation for dominant frequency analysis.

BACKGROUND: Dominant frequency (DF) analysis of atrial electrograms has become an important method in characterizing atrial fibrillation (AF). As a classic method, Botteron's approach is widely used in the preprocessing of frequency analysis during AF. It includes three steps: (1) band-pass filtering at 40-250 Hz, (2) absolute value, and (3) low-pass filtering at 20 Hz. This paper aims to expound the necessity and adjustability of each step. METHODS AND RESULTS: Unipolar epicardial mapping signals were recorded during AF from eight mongrel dogs with cholinergic AF model. Episodes of these data were randomly selected to evaluate the impact of different pass bands and the necessity of low-pass filtering with 20 Hz cutoff frequency. Each episode of AF signal is 5 s long with a sampling rate of 2 kHz. Simulated electrograms were adopted to discuss the role of taking absolute value. Furthermore, direct spectral analysis method (FFT et al.) is compared with Botteron's preprocessing approach. According to our statistical analysis, the pass band of 40-250 Hz was not the best, while 20-100 Hz presented the high accuracy rate of DF. From the comparing result of direct FFT without Botteron's approach we deduced that the rectification of absolute value was meaningful for the fundamental atrial frequency. The final step, 20 Hz low-pass filter can completely be omitted in DF analysis. In consideration of the demand for real-time distribution of DF in clinical or experimental situations, down-sampling method and the impact of ventricular artifacts on DF was also discussed. CONCLUSION: In the actual application of the three preprocessing steps, the pass band selection of band-pass filter can be adjusted and the rectification of taking absolute value is important. Nevertheless, the final step of 20 Hz low-pass filter is totally unnecessary. In real-time signal processing situations, taking down-sampling method and ignoring the ventricular artifacts can also have high performance in DF analysis of atrial electrograms.

Lewis Lung Cancer Cells Promote SIGNR1(CD209b)-Mediated Macrophages Polarization Induced by IL-4 to Facilitate Immune Evasion.

Tumor-associated macrophages are a prominent component of lung cancer and contribute to tumor progression by facilitating the immune evasion of cancer cells. DC-SIGN (CD209) assists in the immune evasion of a broad spectrum of pathogens and neoplasms by inhibiting the maturation of DCs and subsequent cytokines production. However, the expression of DC-SIGN in macrophages and its role in mediating immune evasion in lung cancer and the underlying mechanism remain unclear. Our study aimed to identify the immunosuppressive role of SIGNR1 in murine macrophage differentiation and lung cancer progression. We found that SIGNR1-positive RAW264.7 macrophages were enriched in mixed cultures with Lewis lung cancer cells (LLC) (ratio of RAW 264.7 to LLC being 1:1) after stimulation with IL-4. Moreover, LLC-educated macrophages exhibited significantly higher levels of IL-10 but lower IL-12 in response to IL-4 treatment as determined by RT-PCR and ELISA. However, inhibition of SIGNR1 markedly hampered the production of IL-10, indicating that SIGNR1 was indispensable for IL-4+LLC induced macrophage polarization towards the M2 subtype. Furthermore, polarized M2 cells immersed in a tumor microenvironment promoted the migration of LLCs, as measured by transwell assays, but migration was suppressed after blockade of SIGNR1 using CD209b antibody. In addition, IL-4+LLC-educated macrophages reduced the proliferation of the activated T cells and reduced IFN-γ-mediated Th1 response in T cells, while SIGNR1 inhibition rescued Th1 cell functions. In conclusion, murine SIGNR1 expressed in LLC-educated macrophages appears to mediate IL-4-induced RAW264.7 macrophage polarization and thus facilitate lung cancer evasion.