Nirmatrelvir/ritonavir treatment of patients with COVID-19 taking tacrolimus: case series describing the results of drug-drug interactions.

Nirmatrelvir/ritonavir is a novel drug combination that is authorized by the Food and Drug Administration for the treatment of coronavirus disease 2019 (COVID-19). Ritonavir is a cytochrome P450 3A inhibitor and a P-glycoprotein inhibitor that increases the plasma concentration of tacrolimus and other medications. We describe the cases of two patients treated with nirmatrelvir/ritonavir: a patient who had undergone kidney transplantation and another with a history of hematopoietic stem cell transplantation. Toxic concentrations of tacrolimus were induced in both. This case series highlights the risk associated with the concomitant administration of tacrolimus and nirmatrelvir/ritonavir.

Construction and Application of Fluorescent Probes with Imine Protective Groups for Hypochlorite Detection.

Several fluorescent probes have been designed to detect ClO- in biological systems based on the isomerization mechanism of C = N bonds. Particularly, fluorescein has emerged as an important fluorophore for detecting ClO- because of its unique properties. Previously, we introduced the fluorescein analog F-1 with an active aldehyde group. In this study, two ClO- fluorescent sensors (F-2 and F-3) with imine groups were designed and synthesized using diaminomaleonitrile and 2-hydrazylbenzothiazole as amines. The electron cloud distribution of F-2 and F-3 in ground and excited states was explored via Gaussian calculations, reasonably explaining their photophysical properties. The fluorescence detection of ClO- in solution using the two probes (F-2 and F-3) was realized based on the mechanism of imine deprotection with ClO-. NaClO concentration titration demonstrated that the colorimetric detection of ClO- with the naked eye could be achieved using both F-2 and F-3. However, after adding ClO-, the fluorescence intensity of probe F-2 increased, whereas that of probe F-3 first decreased and then increased. Probes F-2 and F-3 exhibited good selectivity, anti-interference capability, and sensitivity, with the detection limits of 169.95 and 37.30 µM, respectively. Owing to their low cell toxicity, probes F-2 and F-3 can be applied to detect ClO- in vivo. The design approach adopted in this study will further advance the future development of ClO- chemical probes through the removal of C = N bond isomerization.

FBXW7 loss of function promotes esophageal squamous cell carcinoma progression via elevating MAP4 and ERK phosphorylation.

BACKGROUND: Increasing evidence suggests that FBXW7 has a high frequency of mutations in esophageal squamous cell carcinoma (ESCC). However, the function of FBXW7, especially the mutations, is not clear. This study was designed to investigate the functional significance of FBXW7 loss of function and underlying mechanism in ESCC. METHODS: Immunofluorescence was applied to clarify the localization and main isoform of FBXW7 in ESCC cells. Sanger sequencing were performed to explore mutations of FBXW7 in ESCC tissues. Proliferation, colony, invasion and migration assays were performed to examine the functional roles of FBXW7 in ESCC cells in vitro and in vivo. Real-time RT-PCR, immunoblotting, GST-pulldown, LC-MS/MS and co-immunoprecipitation assay were used to explore the molecular mechanism underlying the actions of FBXW7 functional inactivation in ESCC cells. Immunohistochemical staining were used to explore the expression of FBXW7 and MAP4 in ESCC tissues. RESULTS: The main FBXW7 isoform in ESCC cells was the ß transcript in the cytoplasm. Functional inactivation of FBXW7 led to activation of the MAPK signaling pathway and upregulation of the downstream MMP3 and VEGFA, which enhanced tumor proliferation cell invasion and migration. Among the five mutation forms screened, S327X (X means truncated mutation) had an effect similar to the FBXW7 deficiency and led to the inactivation of FBXW7 in ESCC cells. Three other point mutations, S382F, D400N and R425C, attenuated but did not eliminate FBXW7 function. The other truncating mutation, S598X, which was located outside of the WD40 domain, revealed a tiny attenuation of FBXW7 in ESCC cells. Notably, MAP4 was identified as a potential target of FBXW7. The threonine T521 of MAP4, which was phosphorylated by CHEK1, played a key role in the FBXW7-related degradation system. Immunohistochemical staining indicated that FBXW7 loss of function was associated with tumor stage and shorter survival of patients with ESCC. Univariate and multivariate Cox proportional hazards regression analyses showed that high FBXW7 and low MAP4 was an independent prognostic indicator and prospective longer survival. Moreover, a combination regimen that included MK-8353 to inhibit the phosphorylation of ERK and bevacizumab to inhibit VEGFA produced potent inhibitory effects on the growth of FBXW7 inactivation xenograft tumors in vivo. CONCLUSIONS: This study provided evidence that FBXW7 loss of function promoted ESCC via MAP4 overexpression and ERK phosphorylation, and this novel FBXW7/MAP4/ERK axis may be an efficient target for ESCC treatment.

ANXA2 as a novel substrate of FBXW7 promoting esophageal squamous cell carcinoma via ERK phosphorylation.

Our recent study suggests that FBXW7 loss of function plays a critical function in esophageal cancer. However, the mechanism of FBXW7 in promoting esophageal cancer is still unclear. Here, we explored the interaction protein of FBXW7 by screening of GST-pulldown and LC-MS/MS analysis in esophageal squamous cell carcinoma (ESCC) and identified ANXA2 as a potential target of FBXW7. FBXW7 loss of function could restore the expression of ANXA2 and promote the malignant biological characteristics of ESCC cells in vitro. Up-regulation of ANXA2 enhances the ERK pathway in ESCC. Furthermore, the 23rd tyrosine residue of ANXA2, phosphorylated by SRC, was regarded as playing important roles in the FBXW7-related degradation system. In clinical samples, we found that ANXA2 had high expression in ESCC tissues. High ANXA2 was associated with poor tumor staging. More importantly, we designed a combination regimen including SCH779284, a clinical ERK inhibitor against the phosphorylation of EKR and siRNA targeting ANXA2 by intratumor injection, and it produced potent inhibitory effects on the growth of xenograft tumors in vivo. In conclusion, this study provided evidence that FBXW7 loss of function could promote esophageal cancer through ANXA2 overexpression, and this novel regulation pathway may be used as an efficient target for ESCC treatment.

Multidisciplinary Guidelines for the Rational Use of Topical Non-Steroidal Anti-Inflammatory Drugs for Musculoskeletal Pain (2022).

(1) Background: Topical non-steroidal anti-inflammatory drugs (NSAIDs) are one of the primary drugs for treating musculoskeletal pain. However, there are currently no evidence-based recommendations about drug selection, drug administration, drug interactions, and use in special populations or other pharmacology-related content of such medications. To this end, the Chinese Pharmaceutical Association Hospital Pharmacy Professional Committee developed multidisciplinary guidelines on using topical NSAIDs to treat musculoskeletal pain. (2) Methods: The guidelines development process followed the World Health Organization guideline development handbook, the GRADE methodology, and the statement of Reporting Items for Practice Guidelines in Healthcare. The guideline panel used the Delphi method to identify six clinical questions to be addressed in the guidelines. An independent systematic review team conducted a systematic search and integration of evidence. (3) Results: Based on the balance between the benefits and harms of an intervention, the quality of the evidence, patient preferences and values, and resource utilization, the guideline panel developed 11 recommendations and nine expert consensuses on using topical NSAIDs to treat acute and chronic musculoskeletal pain. (4) Conclusions: Based on the effectiveness and overall safety of topical NSAIDs, we recommend patients with musculoskeletal pain use topical NSAIDs and suggest high-risk patients use topical NSAIDs, such as those with other diseases or receiving other concurrent treatments. The evidenced-based guidelines on topical NSAIDs for musculoskeletal pain incorporated a pharmacist perspective. The guidelines have the potential to facilitate the rational use of topical NSAIDs. The guideline panel will monitor the relevant evidence and update the recommendations accordingly.

Databases, Knowledgebases, and Software Tools for Virus Informatics.

Virus infection is a common social health issue. In the past decades, serious virus infectious events have caused great loss in people's life and the economics. The nature of rapid widespread and frequent variation increases the difficulty for precision viral prevention and treatment. In the era of big data and artificial intelligence (AI), advances in bioinformatics techniques bring unprecedented opportunities for virus informatics study, which contribute to the systems-level modeling of virus biology. In this chapter, data resources including virus-related databases and knowledgebases are introduced. Bioinformatics models and software tools for multiple sequence alignment, evolutionary analysis, and genome-wide research of viruses are summarized and emphasized. Translational applications of recently developed data-driven and AI-assisted methods to viral cases such as SARS-CoV-2, HBV/HCV, and influenza virus are discussed. Finally, the concept and significance of virus informatics are highlighted for both virus surveillance and health promotion.

Investigation on fluorescein derivatives with thermally activated delayed fluorescence and their applications in imaging.

Fluorescein derivatives with thermally activated delayed fluorescence (TADF) show much stronger competition ability and vaster prospects than traditional fluorescein dyes due to their prominent long lifetime. It will be of great significance to synthesize more fluorescein derivatives with TADF. In this work, compounds DCF-MPYA and FL with TADF properties were obtained by fine tuning the substituents' structure on the basis of fluorescein derivative DCF-MPYM. Their long-lived triplet excited states (21.78 µs, 32.0 µs) were proved by nanosecond time-resolved transient difference absorption spectra. The steady-state and time-resolved fluorescence spectra showed that DCF-MPYA and FL exhibited red fluorescence around 645 nm and 651 nm, respectively. The results of sensitivity to oxygen and heavy atoms further demonstrated that the time-resolved fluorescence spectra originate from the delayed fluorescence. The time correlated single-photon counting (TCSPC) data indicated that DCF-MPYA and FL showed long-lived lifetimes of 13.16 µs and 23.72 µs, respectively. The energy gap (ΔE ST) between the singlet (S1) and triplet (T1) states of DCF-MPYA and FL was calculated to be 3.32 meV and 9.98 meV from the decay rate of DF as a function of temperature. The small energy gap is conducive to the occurrence of efficient TADF at room temperature. Meanwhile, Gaussian calculation was employed to observe the electron density of DCF-MPYA and FL in the ground and excited states. The calculation results indicate that the shapes and energy levels of the highest occupied molecular orbitals (HOMOs), lowest unoccupied molecular orbitals (LUMOs), and LUMOs+1 for the monoanion and dianion forms are similar and thus DCF-MPYA and FL exhibit almost the same luminescence properties. Finally, DCF-MPYA and FL with low toxicity were used in confocal and time-resolved fluorescence imaging. Our construction strategy will be beneficial for developing more fluorescein derivatives with TADF in the future.

Nickel (ii) effects on Anammox reaction: reactor performance, dehydrogenase, sludge morphology and microbial community changes.

Nickel (ii) (Ni2+) is considered as one of the necessary trace elements in the process of Anammox culture, but it may have toxic effects at high concentration. This study explored the long-term influence of Ni2+ on the denitrification efficiency of Anammox bioreactors. The results showed that when the concentration of Ni2+ was 0.5 mg/L, the bioreactor had the highest denitrification efficiency, while the removal efficiency of ammonia nitrogen and nitrite nitrogen gradually decreased at concentrations higher than 2 mg/L, and the removal rates of ammonia nitrogen and nitrite nitrogen were 26% and 39.81% at the end of the experiment, respectively. The NRR was decreased from 7.47 kg N/m3 d to 3.28 kg N/m3 d during the whole process. The highest concentration of microbial dehydrogenase was attained in about 40 days; in the meantime, its ability to consume organic matter was also maximized. The sludge morphology was changed from granular cluster to loose flocculant with a small number of spherical and filamentous bacteria and bacilli distributed on the surface. At the end of the experiment, both species richness and community diversity were reduced, and the proportion of the dominant bacteria Candidatus Kuenenia was also decreased from 59.89% to 36.72%.

TIPE3 promotes non-small cell lung cancer progression via the protein kinase B/extracellular signal-regulated kinase 1/2-glycogen synthase kinase 3ß-ß-catenin/Snail axis.

BACKGROUND: Tumor necrosis factor-α-induced protein 8-like 3 (TNFAIP8L3, also called TIPE3) has been shown to activate PI3K-AKT and MEK-ERK pathways. However, the roles of TIPE3 in progression of lung cancer are largely unknown. METHODS: Immunohistochemistry and western blotting were carried out to analyze the expression of TIPE3 in lung cancer clinical tissues and cells. TIPE3-overexpressing and knock-down NSCLC cell lines were established by transfer of TIPE3 coding sequence and shRNA, respectively. In vitro functional assays were performed to assess the effects of TIPE3 on proliferation and metastasis of NSCLC cells. Tumor xenograft mouse model was used to examine the roles of TIPE3 in growth of NSCLC cells in vivo. Western blotting, immunofluorescence, and immunohistochemistry were conducted to evaluate the association of TIPE3 and molecules related to AKT/ERK1/2-GSK3ß-ß-catenin/Snail pathway. PI3K, MEK, or GSK3ß kinase and proteasome inhibition assays as well as ß-Trcp and STUB1 siRNA assays were employed to determine the contribution of AKT/ERK1/2-GSK3ß signaling and ubiquitin-proteasome pathway to the regulatory effects of TIPE3 on expression of ß-catenin, Snail1, and Slug. RESULTS: We demonstrated that TIPE3 was elevated in lung cancer tissues and cells. The expression level of TIPE3 was positively correlated with malignant clinicopathological characteristics of lung cancer patients, such as tumor size, pathologic stage, and lymph node metastasis. Knockdown of TIPE3 suppressed the proliferation and growth of NSCLC cells as well as their migration and invasion ability, whereas TIPE3 overexpression facilitated these biological processes. Mechanistic data showed that TIPE3 promoted AKT and ERK1/2 signaling, inactivated GSK3ß activity, and enhanced the expression and transcriptional activity of ß-catenin, Snail1, and Slug in NSCLC cells. Kinase or proteasome inhibition and ß-Trcp or STUB1 knockdown assays further revealed that TIPE3 upregulated ß-catenin, Snail1, and Slug via the AKT/ERK1/2-GSK3ß pathway, in an ubiquitin-proteasome-dependent manner. More importantly, clinical data demonstrated that the expression level of TIPE3 was positively associated with the activation of AKT/ERK1/2-GSK3ß-ß-catenin/Snail pathway in lung cancer. CONCLUSIONS: Our findings indicate that upregulation of TIPE3 promotes the progression of human NSCLC considerably by activating ß-catenin, Snail1, and Slug transcriptional signaling via the AKT/ERK1/2-GSK3ß axis. Therefore, TIPE3 may represent a potential therapeutic target for NSCLC.

Sirtuin 7 promotes non­small cell lung cancer progression by facilitating G1/S phase and epithelial­mesenchymal transition and activating AKT and ERK1/2 signaling.

Increasing evidence has indicated the roles of sirtuin 7 (SIRT7) in numerous human cancers. However, the effects and the clinical significance of SIRT7 in human lung cancer is largely unknown. The present research demonstrated that SIRT7 was increased in human lung cancer tumor tissues. SIRT7 upregulation was associated with clinicopathological characteristics of lung cancer malignancy including positive lymph node metastasis, high pathologic stage and large tumor size. SIRT7 was also upregulated in human non­small cell lung cancer (NSCLC) cell lines. Furthermore SIRT7­overexpressed A549 (A549­SIRT7) and SIRT7­knocked down H292 (H292­shSIRT7) human NSCLC cell lines were established. Using these NSCLC cells and xenograft mouse models, it was revealed that SIRT7 overexpression markedly promoted growth and G1 to S cell cycle phase transition as well as migration, invasion and distant lung metastasis in A549 NSCLC cells, whereas SIRT7 knockdown suppressed these processes in H292 NSCLC cells. Mechanistically, in A549 NSCLC cells, SIRT7 overexpression significantly activated not only protein kinase B (AKT) signaling but also extracellular signal­regulated kinase 1/2 (ERK1/2) signaling. SIRT7 overexpression also significantly downregulated cyclin­dependent kinase (CDK) inhibitors including p21 and p27 as well as upregulated cyclins including cyclin D1 and cyclin E1, and CDKs including CDK2 and CDK4. Notably, the epithelial­mesenchymal transition (EMT) process of A549 NSCLC cells was facilitated by SIRT7 overexpression, as evidenced by E­cadherin epithelial marker downregulation and mesenchymal markers (N­cadherin, vimentin, Snail and Slug) upregulation. In addition, SIRT7 knockdown in H292 NSCLC cells exhibited the opposite regulatory effects. Moreover, inhibition of AKT signaling abated the promoting effects of SIRT7 in NSCLC cell proliferation and EMT progression. The present data indicated that SIRT7 accelerated human NSCLC cell growth and metastasis possibly by promotion of G1 to S­phase transition and EMT through modulation of the expression of G1­phase checkpoint molecules and EMT markers as well as activation of AKT and ERK1/2 signaling. SIRT7 could be an innovative potential target for human NSCLC therapy.

Discovery and characterization of long intergenic non-coding RNAs (lincRNA) module biomarkers in prostate cancer: an integrative analysis of RNA-Seq data.

BACKGROUND: Prostate cancer (PCa) is a leading cause of cancer-related death of men worldwide. There is an urgent need to develop novel biomarkers for PCa prognosis and diagnosis in the post prostate-specific antigen era. Long intergenic noncoding RNAs (lincRNAs) play essential roles in many physiological processes and can serve as alternative biomarkers for prostate cancer, but there has been no systematic investigation of lincRNAs in PCa yet. RESULTS: Nine lincRNA co-expression modules were identified from PCa RNA-Seq data. The association between the principle component of each module and the PCa phenotype was examined by calculating the Pearson's correlation coefficients. Three modules (M1, M3, and M5) were found associated with PCa. Two modules (M3 and M5) were significantly enriched with lincRNAs, and one of them, M3, may be used as a lincRNA module-biomarker for PCa diagnosis. This module includes seven essential lincRNAs: TCONS_l2_00001418, TCONS_l2_00008237, TCONS_l2_00011130, TCONS_l2_00013175, TCONS_l2_00022611, TCONS_l2_00022670 and linc-PXN-1. The clustering analysis and microRNA enrichment analysis further confirmed our findings. CONCLUSION: The correlation between lincRNAs and protein-coding genes is helpful for further exploration of functional mechanisms of lincRNAs in PCa. This study provides some important insights into the roles of lincRNAs in PCa and suggests a few lincRNAs as candidate biomarkers for PCa diagnosis and prognosis.