Case report: EGFR-TKI rechallenge after osimertinib-induced interstitial lung disease: a case report and literature review.

Background: Osimertinib, a third-generation tyrosine kinase inhibitor (TKI), has demonstrated significant efficacy in treating non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations. However, EGFR-TKI-induced interstitial lung disease (ILD), a well-known adverse effect, can seriously affect the treatment outcome. There is currently no international consensus on the efficacy and safety of re-administration of EGFR-TKI after EGFR-TKI-induced ILD. Case summary: We report a case of a 62-year-old male with stage IV lung adenocarcinoma and EGFR L858R mutation who was treated with osimertinib at a dose of 80 mg/day as first-line therapy. On the sixth day of treatment, the patient developed grade 4 ILD, chest tightness, shortness of breath, and paroxysmal dry cough. Arterial blood gas analysis indicated the presence of type I respiratory failure, while the chest CT scan revealed newly developed ground-glass opacities in both lungs and a considerable amount of pleural effusion on the left side. Subsequently, the patient was administered methylprednisolone for anti-inflammatory therapy, in conjunction with oxygen therapy, anti-infection treatment, and closed thoracic drainage, which resulted in a favourable recovery and discharge after 18 days. During this period, the patient adhered to third-generation EGFR-TKI oral targeted therapy. Nevertheless, within a week of discharge, the patient was readmitted due to the recurrence of chest tightness and shortness of breath. A chest CT scan indicated a recurrent ILD. Despite the administration of high-dose methylprednisolone for 9 days, the patient's condition continued to deteriorate, ultimately resulting in death. Conclusion: It is of the utmost importance to conduct a meticulous evaluation of the severity of osimertinib-induced ILD in order to ascertain the potential risks and benefits of EGFR-TKI rechallenge. Particularly, for patients with grade 4 ILD, firm drug discontinuation should be considered.

Local-field engineering in slot waveguide for fabricating on-chip Bragg grating filters with high reflectivity across a flat broadband.

On-chip Bragg gratings with high reflectivities have been found to have widespread applications in filters, resonators, and semiconductor lasers. However, achieving strong Bragg reflections with flat response across a broad bandwidth on the popular 220 nm silicon-on-insulator (SOI) platform still remains a challenge. In this paper, such a high performance device is proposed and fabricated, which is based on a slot waveguide with gratings etched on the inner sidewalls of the slot. By manipulating the local field in the slot region using a chirped and tapered grating-based mode transition, the device achieves a flat response with ultra-high reflection and low transmission for the TE mode across a broad operating bandwidth. Leveraging the ultra-high birefringence of the SOI waveguide, the device functions both as a TE slot waveguide reflector and a TM pass polarizer. Simulation results demonstrate that the device exhibits an ultra-high rejection of more than 50 dB and a reflectivity exceeding 0.99 for the TE mode across a 91 nm wavelength range, while maintaining a high transmittance of larger than 0.98 for the TM mode. Experimental results validate that the device performance is consistent with the simulation results. A fabricated device based on such a gratings exhibits a low insertion loss (<0.8 dB) and high polarization extinction ratio (>30 dB) over 100 nm bandwidth (1484 nm-1584 nm), demonstrating that the performance of the present design is competitive with that of the state-of-the-art SOI Bragg gratings.

All-silicon TM polarizer covering the 1260-1675†nm bandwidth using a band engineered subwavelength grating waveguide.

A TM polarizer working for whole optical communication bands with high performance is proposed on a 220-nm-thick silicon-on-insulator (SOI) platform. The device is based on polarization-dependent band engineering in a subwavelength grating waveguide (SWGW). By utilizing an SWGW with a relatively larger lateral width, an ultra-broad bandgap of ∼476 nm (1238 nm-1714nm) is obtained for the TE mode, while the TM mode is well supported in this range. Then, a novel tapered and chirped grating design is adopted for efficient mode conversion, which results in a polarizer with a compact footprint (3.0 µm × 18 µm), low insertion loss (IL < 1.15 dB) and high polarization extinction ratio (PER > 21 dB) covering O-U bands (1260 nm-1675 nm). Experimental results show that the fabricated device has an IL < 1.0 dB and PER > 22 dB over a 300- nm bandwidth, which is limited by our measurement setup. To the best of our knowledge, no TM polarizer on the 220-nm SOI platform with comparable performance covering O-U bands has ever been reported.

Glycyrrhetinic acid ameliorates diosbulbin B-induced hepatotoxicity in mice by modulating metabolic activation of diosbulbin B.

Exposure to diosbulbin B (DBB), the primary component of the herbal medicine Dioscorea bulbifera L. (DB), can cause liver injury in humans and experimental animals. A previous study found DBB-induced hepatotoxicity was initiated by CYP3A4-mediated metabolic activation and subsequent formation of adducts with cellular proteins. The herbal medicine licorice (Glycyrrhiza glabra L.) is frequently combined with DB used in numerous Chinese medicinal formulas in an effort to protect against DB-elicited hepatotoxicity. Importantly, glycyrrhetinic acid (GA), the major bioactive ingredient in licorice, inhibits CYP3A4 activity. The study aimed to investigate the protection of GA against DBB-induced hepatotoxicity and the underlying mechanism. Biochemical and histopathological analysis showed GA alleviated DBB-induced liver injury in a dose-dependent manner. In vitro metabolism assay with mouse liver microsomes (MLMs) indicated that GA decreased the generation of metabolic activation-derived pyrrole-glutathione (GSH) conjugates from DBB. Toxicokinetic studies demonstrated that GA increased maximal serum concentration (Cmax ) and area under the serum-time curve (AUC) of DBB in mice. In addition, GA attenuated hepatic GSH depletion caused by DBB. Further mechanistic studies showed that GA reduced the production of DBB-derived pyrroline-protein adducts in a dose-dependent manner. In conclusion, our findings demonstrated that GA exerted protective effect against DBB-induced hepatotoxicity, mainly correlated with suppressing the metabolic activation of DBB. Therefore, the development of a standardized combination of DBB with GA may protect patients from DBB-induced hepatotoxicity.

Highly scalable and flexible on-chip all-silicon mode filter using backward mode conversion gratings.

Mode filters are fundamental elements in a mode-division multiplexing (MDM) system for reducing modal cross-talk or realizing modal routing. However, the previously reported silicon mode filters can only filter one specific mode at a time and multiple modes filtering usually needs a cascade of several filters, which is adverse to highly integrated MDM systems. Here, we propose a unique concept to realize compact, scalable and flexible mode filters based on backward mode conversion gratings elaborately embedded in a multimode waveguide. Our proposed method is highly scalable for realizing a higher-order-mode-pass or band-mode-pass filter of any order and capable of flexibly filtering one or multiple modes simultaneously. We have demonstrated the concept through the design of four filters for different order of mode(s) and one mode demultiplexer based on such a filter, and the measurement of two fabricated 11µm length filters (TE1-pass/TE2-pass) show that an excellent performance of insertion loss <1.0dB/1.5dB and extinction ratio >29dB/28.5dB is achieved over a bandwidth of 51.2nm/48.3nm, which are competitive with the state-of-the-art.

CT imaging features regarding ground-glass nodules and solid lesions reflect prognostication of synchronous multiple lung adenocarcinoma.

The prognosis of synchronous multiple lung adenocarcinoma (SMLA) dramatically differs due to its nature of multiple primaries or intrapulmonary metastases. This study aimed to assess computed tomography (CT)-reflected SMLA features regarding ground-glass nodules (GGNs) and solid lesions and their correlation with prognostication. One seventy eight SMLA patients who underwent surgical resection were reviewed. According to preoperative CT features, patients were categorized as: multiple GGN (MG) group: MGs without solid lesions; solid plus GGN (SPG) group: one solid lesion and at least one GGN; multiple solid (MS) group: MS lesions, with or without GGNs. Clinical characteristics, disease-free survival (DFS), and overall survival (OS) were retrieved. Largest tumor size (P < .001) and lymph-node metastasis prevalence (P < .001) were different among three groups, which were highest in the MS group, followed by the SPG group, and lowest in the MG group. Besides, the dominant tumor subtype also varied among the three groups (P < .001), while no difference in other clinical characteristics was discovered. DFS was more deteriorative in the MS group compared to the SPG group (P = .017) and MG group (P < .001), while of no difference between the SPG group and MG group (P = .128). Meanwhile, OS exhibited similar treads among the three groups. Besides, after multivariate Cox analyses adjustment, MS versus MG independently correlated with DFS (P = .030) and OS (P = .027), but SPG versus MG did not. In conclusion, preoperative CT-imaging MS lesions reflect advanced disease features and poor prognosis compared to MG and solid lesion plus GGN in SMLA patients who underwent surgical resection.

YTHDF2 Regulates Macrophage Polarization through NF-κB and MAPK Signaling Pathway Inhibition or p53 Degradation.

Macrophages are heterogeneous cells that can be polarized into M1 or M2 phenotype. m6A "reader" YTH domain family protein 2 (YTHDF2) has been the m6A binding protein with the highest activity, which can recognize and disturb m6A-containing mRNA in processing bodies to reduce mRNA stability. YTHDF2 is recently identified as an effective RNA binding protein that modulates inflammatory gene levels within inflammatory responses. However, the role of YTHDF2 in M1/M2 macrophage polarization has not been reported. We established a M1/M2 macrophage polarization model using bone-marrow-derived macrophages and found that the expression levels of YTHDF2 in M1/M2 macrophages were both elevated. YTHDF2-knockdown macrophage polarization model was then established, and through qPCR, ELISA, and FACS, we discovered that suppressing YTHDF2 encouraged M1 polarization but restrained M2 polarization. In M1 macrophages, YTHDF2 silencing had no significant effect on p53 expression; however, in YTHDF2 knockdown, M2 macrophage p53 expression was remarkably upregulated. p53 inhibitor PFT-α was then applied and revealed that suppressing p53 simultaneously promoted YTHDF2-silenced M1 polarization and facilitated M2 macrophage polarization. Actinomycin D assays were further utilized to examine the mRNA degradation level of different cytokines, and p53 mRNA degradation in YTHDF2-depleted M2 cells was discovered impeded. Western Blot analysis also implied that a deficit in YTHDF2 expression may activate MAPK and NF-κB pathways. In this study, YTHDF2 induces M2 macrophage polarization by promoting the degradation of p53 mRNA. YTHDF2 suppresses M1 macrophage polarization by inhibiting NF-κB, p38, and JNK signaling pathways, yet p53 remains unaffected in YTHDF2-silenced M1 macrophages.

The Efficacy of Fosbretabulin Disodium Combined with Radiofrequency Ablation in Lung Cancer.

Radiofrequency ablation (RFA) is a technology that uses radiofrequency thermal effect to induce coagulation necrosis of tumor tissue under the guidance of imaging. However, distant metastasis of tumor cells caused by tumor angiogenesis can lead to incomplete tumor clearing. In this study, LLC1 cell line was used for the construction of subcutaneous xenografts. Either 10 mg/kg or 20 mg/kg Fosbretabulin disodium (FBTD) was intragastrically administered every 2 days for a week. RFA was performed at the end of medication. The proportion of T cells was examined by flow cytometry. Serum IgG and IgA levels of mice were examined by ELISA. Expression of certain genes was estimated by qRT-PCR assay. In this study, we demonstrated that FBTD was able to significantly enhance RFA-induced immune function in tumor-bearing mice by upregulating RFA-induced CD8+ killer T cells. Consistently, 10 mg/kg or 20 mg/kg FBTD therapy upregulated the percentage of IFNγ+ and TNFα+ CD8+ tumor infiltrating lymphocytes in tumor-bearing mice compared to the RFA alone or FBTD alone group. Mechanistically, we reported that FBTD inhibited the RFA-induced PD-1 and PD-L1 upregulation in vivo. In conclusion, we demonstrated that FBTD promoted the antitumor effects of RFA in lung tumor-bearing mice in this study.

Correlation Analysis of Computed Tomography Features and Pathological Types of Multifocal Ground-Glass Nodular Lung Adenocarcinoma.

To investigate the correlation between computed tomography (CT) image characteristics of multiple lung ground-glass nodules (GGNs) and pathological classification, the CT image data of multiple lung GGN patients confirmed by pathology (n = 132) in our hospital were collected. The imaging features of GGNs were analyzed by qualified physicians, including lesion size (diameter, volume, and mass), location, CT values (mean and relative CT values), lesion morphology (round and irregular), marginal structure (pagination and burr), internal structure (bronchial inflation sign), and adjacent structure (pleural depression). CT imaging analysis was performed for the subtype of infiltrating adenocarcinoma (IAC). In CT findings, GGNs were greatly different from adenomatous hyperplasia (AAH), pure GGN adenocarcinoma in situ (AIS), and microinvasive adenocarcinoma (MIA) in terms of marginal structure, lesion morphology, internal structure, adjacent structure, and size (P < 0.05). The mean and relative CT values of mural adenocarcinoma, acinar adenocarcinoma, and papillary adenocarcinoma of IAC subtypes were greatly different from those of AAH/AIS/MIA (P < 0.05). In summary, the CT images of GGNs can be used as the basis for the differentiation of AAH, AIS, and MIA early noninvasive types and IAC invasive types, and the CT value of the IAC subtype can be used as the basis for the classification and differentiation of IAC pathological subtypes.

The prevalence and clinical features of pulmonary embolism in patients with AE-COPD: A meta-analysis and systematic review.

BACKGROUND: The prevalence of pulmonary embolism (PE) in the acute exacerbation of chronic obstructive pulmonary disease (AE-COPD) is highly controversial. We conducted a systematic review and meta-analysis to summarize the epidemiology and characteristics of PE with AE-COPD for current studies. METHODS: We searched the PubMed, EMBASE, Cochrane Library and Web of Science databases for studies published prior to October 21, 2020. Pooled proportions with 95% confidence intervals (95% CIs) were calculated using a random effects model. Odds ratios (ORs) and mean differences (MDs) with 95% confidence intervals were used as effect measures for dichotomous and continuous variables, respectively. RESULTS: A total of 17 studies involving 3170 patients were included. The prevalence of PE and deep vein thrombosis (DVT) in AE-COPD patients was 17.2% (95% CI: 13.4%-21.3%) and 7.1% (95% CI: 3.7%-11.4%%), respectively. Dyspnea (OR = 6.77, 95% CI: 1.97-23.22), pleuritic chest pain (OR = 3.25, 95% CI: 2.06-5.12), lower limb asymmetry or edema (OR = 2.46, 95% CI:1.51-4.00), higher heart rates (MD = 20.51, 95% CI: 4.95-36.08), longer hospital stays (MD = 3.66, 95% CI: 3.01-4.31) were associated with the PE in the AE-COPD patients. Levels of D-dimer (MD = 1.51, 95% CI: 0.80-2.23), WBC counts (MD = 1.42, 95% CI: 0.14-2.70) were significantly higher and levels of PaO2 was lower (MD = -17.20, 95% CI: -33.94- -0.45, P<0.05) in the AE-COPD with PE group. The AE-COPD with PE group had increased risk of fatal outcome than the AE-COPD group (OR = 2.23, 95% CI: 1.43-3.50). CONCLUSIONS: The prevalence of PE during AE-COPD varies considerably among the studies. AE-COPD patients with PE experienced an increased risk of death, especially among the ICU patients. Understanding the potential risk factors for PE may help clinicians identify AE-COPD patients at increased risk of PE. PROSPERO REGISTRATION NUMBER: CRD42021226568.

Stretchable MXene/Thermoplastic Polyurethanes based Strain Sensor Fabricated Using a Combined Electrospinning and Electrostatic Spray Deposition Technique.

In this work, a novel flexible electrically resistive-type MXene/Thermoplastic polyurethanes(TPU) based strain sensors was developed by a composite process of electrospinning (ES) and electrostatic spray deposition (ESD). Compared with other deposition processes, the sensing layer prepared by ESD has better adhesion to the ES TPU nanofiber membrane and is not easy to crack during the stretching process, thereby greatly improving the working range of the strain sensor. Furthermore, we obtained the sandwich structure easily by ES on the surface of the sensing layer again. This will help make the stress distribution more uniform during the stretching process and further increase the strain sensing range. The ESD-ES strain sensors were attached on skin to monitor various human motions. The results demonstrate that our ESD-ES strain sensors have wide application prospects in smart wearable device.

Roxithromycin attenuates inflammation via modulation of RAGE-influenced calprotectin expression in a neutrophilic asthma model.

BACKGROUND: Roxithromycin (RXM), a macrolide antibiotic, exhibits anti-asthmatic effects, but its specific mechanism of action remains elusive. We evaluated the effects of RXM on airway inflammation, the expression of calprotectin, and the activity of the receptor of advanced glycation end products (RAGE) to determine whether RXM alleviates inflammation by regulating RAGE activation, and thereby calprotectin expression, in neutrophilic asthma. METHODS: Male Brown Norway rats were sensitized with ovalbumin (OVA) and Freund's complete adjuvant (FCA) mixture, followed by OVA challenge to induce neutrophilic asthma. RXM (30 mg/kg) or FPS-ZM1 (RAGE inhibitor, 1.5 mg/kg) was administered 30 min prior to each challenge. The infiltration of airway inflammatory cells and cytokines, as well as the expression of calprotectin and RAGE, was assessed. RESULTS: The expression of airway inflammatory cells and cytokines was found to be significantly elevated in OVA + FCA-induced rats. Increased expression of both calprotectin and RAGE was also detected in OVA + FCA-induced asthma [bronchoalveolar lavage fluid (BALF) calprotectin: 15.07±1.79 vs. 3.86±0.69 ng/mL; serum calprotectin: 20.47±1.64 vs. 9.29±1.31 ng/mL; lung tissue homogenates calprotectin: 28.82±1.01 vs. 12.02±1.38 ng/mg; BALF RAGE: 762.93±36.47 vs. 294.25±45.92 ng/mL; serum RAGE: 906.43±58.95 vs. 505.60±30.16 ng/mL; lung tissue homogenates RAGE: 1,585.24±177.59 vs. 461.53±63.40 ng/mg; all P<0.001]. However, all of these changes were interrupted by RXM and FPS-ZM1. CONCLUSIONS: RXM exerted similar effects as the RAGE inhibitor FPS-ZM1 in terms of reducing airway inflammation and downregulating the expression of calprotectin and RAGE in a neutrophilic asthma model. Our findings provide novel insights into the mechanisms underlying the effect of RXM pretreatment on neutrophilic asthma. Furthermore, FPS-ZM1 may be useful as an intervention specific to the neutrophilic asthma phenotype.

Fabricating cholyglycine-glucose-6-phosphate dehydrogenase conjugates for cholyglycine detection.

To establish cholyglycine (CG) detection via enzyme multiplied immunoassay technique (EMIT), glucose-6-phosphate dehydrogenase (G6PD) was used as a reporter enzyme to prepare hapten-enzyme conjugate. Gel electrophoresis and UV scanning demonstrated that G6PD was successfully labeled with cholyglycine and CG-G6PD conjugate was obtained. Furthermore, the effects of various parameters on the preparation of CG-G6PD conjugates were investigated. Consequently, CG amount, NADH, D-glucose-6-phosphate (G6P), phosphate buffer and the pH, and ionic strength of solution had important effects on the residual activity of CG-G6PD. Moreover, CG amount, the pH, and G6P played important roles in changing CG labeling location on G6PD. Using the CG-G6PD conjugate as test kit, the cholyglycine-EMIT calibration curve was established, which could be employed in clinical detection of cholyglycine. This study provides some valuable information for preparing hapten-G6PD conjugates. This article is protected by copyright. All rights reserved.

Carotid web and ischemic stroke: a CT angiography study.

OBJECTIVES: We sought to explore the prevalence, demographics, clinical and imaging features of the Carotid web (CaW) on CT angiography (CTA) in patients with cryptogenic and non-cryptogenic stroke through a large-scale retrospective study. MATERIALS AND METHODS: A total of 1662 patients with ischemic stroke and had a neck CTA were retrospectively reviewed. An extensive clinical workup was performed to identify patients with cryptogenic stroke. All neck CTA studies were reconstructed and independently evaluated by two experienced radiologists for presence or absence of CaW on the ipsilateral and contralateral to the stroke side. RESULTS: Thirty-three cases of CaW were eventually diagnosed in patients with ischemic stroke, with a prevalence of 2.2% (33/1489) in a hospital-based series. Twenty-six (26/33, 78.8%) cases of CaW were ipsilateral to the stroke side. There are 18 ipsilateral CaWs (18/285, 6.3%) in cryptogenic stroke patients, and eight ipsilateral CaWs (8/1204, 0.7%) in non-cryptogenic stroke, yielding an odds ratio of 10.1. Cryptogenic stroke patients with ipsilateral CaW were relatively young with a higher prevalence of women. The interrater and intrarater agreement on the CTA based diagnosis of CaW were substantial. CONCLUSIONS: Our results demonstrate a strong correlation between the CaW and cryptogenic stroke in large Asian study population. CTA is the imaging modality of choice for detecting CaW.

Draft genome sequence data of Bacillus subtilis strain 9407, isolated from healthy apples in China.

Bacillus subtilis strain 9407 is an endophyte which was isolated from healthy apples from an infested orchard that exhibits strong inhibitory activity against Botryosphaeria dothidea. Whole-genome sequencing of B. subtilis 9407 was performed using the Illumina Hiseq platforms. Here, we report the draft genome sequence of B. subtilis strain 9407 containing 16 scaffolds (4,062,615 bp), 4033 coding sequences, and an average 43.66% G + C content. The genome contains genes responsible for the production of several bioactive secondary metabolites, including the lipopeptides fengycin and surfactin. The genome information will provide fundamental knowledge of the organism. This whole-genome shotgun data has been deposited at DDBJ/EMBL/GenBank under the accession numbers PISO00000000.

PEI-crosslinked lipase on the surface of magnetic microspheres and its characteristics.

Here, PEI@PMMA microspheres were prepared by grafting polyethyleneimine (PEI) on poly(methyl methacrylate) (PMMA) magnetic microspheres and successfully used to immobilize lipase. The results showed that PEI@PMMA microspheres had strongly adsorbed lipase (49.1 mg/g microsphere) via electrostatic attraction. To prevent lipase shedding, the adsorbed lipase was further crosslinked with PEI on microspheres using glutaraldehyde as crosslinker. Consequently, PEI-crosslinked lipase (2.14 U/mg) exhibited 2.6 times and 1.4 times higher activity respectively than the directly covalent lipase (0.82 U/mg) and the crosslinked lipase aggregates (1.57 U/mg), which was close to the activity of adsorbed lipase (2.20 U/mg). Conformational analysis from FTIR spectroscopy showed that PEI-crosslinked lipase retained its natural structure well. And the α-helix structure seemed to play a key role in enhancing lipase activity. Furthermore, the effects of various parameters on crosslinking reaction were investigated. Also, PEI-crosslinked lipase revealed higher pH and thermal stability. The Michaelis constant (Km) was increased and the optimum temperature of lipase was widened observably after crosslinking with PEI on PEI@PMMA magnetic microspheres.

N6-methyladenosine demethylase FTO promotes M1 and M2 macrophage activation.

Macrophage polarization is the driving force of various inflammatory diseases, especially those involved in M1/M2 imbalance. N6-methyladenosine (m6A) is the most prevalent internal mRNA modification in eukaryotes that affects multiple biological processes, including those involved developmental arrest and immune response. However, the role of m6A in macrophage polarization remains unclear. This study found that FTO silencing significantly suppressed both M1 and M2 polarization. FTO depletion decreased the phosphorylation levels of IKKα/ß, IκBα and p65 in the NF-κB signaling pathway. The expression of STAT1 was downregulated in M1-polarized macrophages while the expression of STAT6 and PPAR-γ decreased in M2 polarization after FTO knockdown. The actinomycin D experiments showed that FTO knockdown accelerated mRNA decay of STAT1 and PPAR-γ. Furthermore, the stability and expression of STAT1 and PPAR-γ mRNAs increased when the m6A reader YTHDF2 was silenced. In conclusion, our results suggest that FTO knockdown inhibits the NF-κB signaling pathway and reduces the mRNA stability of STAT1 and PPAR-γ via YTHDF2 involvement, thereby impeding macrophage activation. These findings indicated a previously unrecognized link between FTO and macrophage polarization and might open new avenues for research into the molecular mechanisms of macrophage polarization-related diseases.

Fabricating cholyglycine-glucose-6-phosphate dehydrogenase conjugates for cholyglycine detection.

To establish cholyglycine (CG) detection via enzyme-multiplied immunoassay technique (EMIT), glucose-6-phosphate dehydrogenase (G6PD) was used as a reporter enzyme to prepare hapten-enzyme conjugate. Gel electrophoresis and UV scanning demonstrated that G6PD was successfully labeled with cholyglycine, and CG-G6PD conjugate was obtained. Furthermore, the effects of various parameters on the preparation of CG-G6PD conjugates were investigated. Consequently, CG amount, nicotinamide adenine dinucleotide, d-glucose-6-phosphate (G6P), phosphate buffer and the pH, and ionic strength of solution had important effects on the residual activity of CG-G6PD. Moreover, CG amount, the pH, and G6P played important roles in changing CG labeling location on G6PD. Using the CG-G6PD conjugate as test kit, the cholyglycine-EMIT calibration curve was established, which could be employed in clinical detection of cholyglycine. This study provides some valuable information for preparing hapten-G6PD conjugates.

Absolute configuration determination of asarinin by synchrotron radiation with crystalline sponge method.

The determination of the absolute configuration of natural products still faces many challenges, especially the active pharmaceutical ingredient which is trace, oily and novel structures. Currently, NMR requires chiral reagents in determining the absolute configuration; ECD involves theoretical calculations and requires chromophores. In this study, the absolute configuration of asarinin had successfully identified by using synchrotron radiation with crystalline sponge method and combining MS with NMR. This method could identifying the crystal structure of trace amorphous substances, resolving the problem of absolute configuration of multi-chiral central compounds, and hopefully providing a new idea and approach for structural elucidation of natural products.

METTL3 Regulates Osteoblast Differentiation and Inflammatory Response via Smad Signaling and MAPK Signaling.

Osteoblasts are crucial bone-building cells that maintain bone homeostasis, whereas inflammatory stimuli can inhibit osteogenesis and activate inflammatory response. N6-methyladenosine (m6A) is the most abundant mRNA modification in eukaryotes and plays important roles in multiple biological processes. However, whether m6A modification affects osteoblast differentiation and inflammatory response remains unknown. To address this issue, we investigated the expression of the N6-adenosine methyltransferase METTL3 and found that it was upregulated during osteoblast differentiation and downregulated after LPS stimulation. We then knocked down METTL3 and observed decreased levels of osteogenic markers, ALP activity, and mineralized nodules, as well as Smad1/5/9 phosphorylation, in LPS-induced inflammation. METTL3 knockdown promoted the mRNA expression and stability of negative regulators of Smad signaling, Smad7 and Smurf1, the same regulatory pattern identified when the m6A-binding protein YTHDF2 was silenced. Moreover, METTL3 depletion enhanced proinflammatory cytokine expression and increased the phosphorylation of ERK, p38, JNK, and p65 in MAPK and NF-κB signaling pathways. The increase in cytokine expression was inhibited after MAPK signaling inhibitor treatment. All data suggest that METTL3 knockdown inhibits osteoblast differentiation and Smad-dependent signaling by stabilizing Smad7 and Smurf1 mRNA transcripts via YTHDF2 involvement and activates the inflammatory response by regulating MAPK signaling in LPS-induced inflammation.