A novel selective ERK1/2 inhibitor, Laxiflorin B, targets EGFR mutation subtypes in non-small-cell lung cancer.

Extracellular regulated protein kinases 1/2 (ERK1/2) are key members of multiple signaling pathways, including the ErbB axis. Ectopic ERK1/2 activation contributes to various types of cancer, especially drug resistance to inhibitors of RTK, RAF and MEK, and specific ERK1/2 inhibitors are scarce. In this study, we identified a potential novel covalent ERK inhibitor, Laxiflorin B, which is a herbal compound with anticancer activity. However, Laxiflorin B is present at low levels in herbs; therefore, we adopted a semi-synthetic process for the efficient production of Laxiflorin B to improve the yield. Laxiflorin B induced mitochondria-mediated apoptosis via BAD activation in non-small-cell lung cancer (NSCLC) cells, especially in EGFR mutant subtypes. Transcriptomic analysis suggested that Laxiflorin B inhibits amphiregulin (AREG) and epiregulin (EREG) expression through ERK inhibition, and suppressed the activation of their receptors, ErbBs, via a positive feedback loop. Moreover, mass spectrometry analysis combined with computer simulation revealed that Laxiflorin B binds covalently to Cys-183 in the ATP-binding pocket of ERK1 via the D-ring, and Cys-178 of ERK1 through non-inhibitory binding of the A-ring. In a NSCLC tumor xenograft model in nude mice, Laxiflorin B also exhibited strong tumor suppressive effects with low toxicity and AREG and EREG were identified as biomarkers of Laxiflorin B efficacy. Finally, Laxiflorin B-4, a C-6 analog of Laxiflorin B, exhibited higher binding affinity for ERK1/2 and stronger tumor suppression. These findings provide a new approach to tumor inhibition using natural anticancer compounds.

Well-being Effects of Natural Disasters: Evidence from China's Wenchuan Earthquake.

This study finds that the Wenchuan earthquake in 2008, one of China's most catastrophic earthquakes, substantially decreased victims' subjective well-being even after incorporating the offsetting effects of post-disaster relief programs. This net well-being impact lasted for nearly 10 years and was on average equivalent to a loss of 67% of the average equivalized household income. Although the post-disaster measures largely restored income, health, and employment, they failed to prevent well-being losses due to family dissolution, as reflected in the higher rates of divorce and widowhood after the earthquake. We find that rural populations, older adults, the less educated, and residents without social insurance were more vulnerable to the earthquake shock. This study uses six waves of a nationally representative dataset of China and a difference-in-differences approach to identify the short- and long-term causal well-being effects of the Wenchuan earthquake. Deeper analyses on mechanisms and heterogeneity suggest that post-disaster policies should focus more on aspects beyond economic factors and on the well-being of disadvantaged populations in particular. Supplementary Information: The online version contains supplementary material available at 10.1007/s10902-022-00609-z.

[The third-generation sequencing combined with targeted capture technology for high-resolution HLA typing and MHC region haplotype identification].

The high-resolution and accurate typing of human leukocyte antigen (HLA) is of great significance for the study of tissue matching in organ transplantation and the correlation between HLA and disease. In this study, the peripheral blood of 12 patients with primary hepatocellular carcinoma was used to compare the advantages and disadvantages of the next- and third-generation sequencing technology for high-resolution HLA typing. In addition, probe capture technology was used to capture the MHC region of YH and HeLa standard cell lines, and a primary hepatocellular carcinoma patient. The captured products were sequenced using PacBio platform to assess the potential of ultra-long reads sequencing technology for analysis of the entire MHC region. Our results showed that: (1) the next- and third-generation sequencing technology can both achieve 6-8 digit high resolution in HLA typing. However, the coverage of the third-generation is significantly better than the next-generation sequencing technology. (2) The ultra-long reads of the third generation sequencing can directly span the entire amplicon region, which has obvious advantages for haplotype phasing, with 92.79% of the HLA genes having accurate phasing results, which is much higher than the 75.65% from the next-generation data. (3) The long-reads from the third generating sequencing can not only be used to assemble the MHC region but also the ability to phase the entire MHC region of 3.6 Mb, thereby helping to clarify the localization information of the mutation sites, alleles and non-coding regions on each MHC haplotype, and providing a theoretical basis for the study of immune and other related diseases.

The clinical effect of the digital guide in the early implant restoration of second molars.

BACKGROUND: In the absence of contraindications to implants, implant repair is the preferred method to treat mandibular second molar loss. OBJECTIVE: To compare the clinical effects of a traditional implant guide and digital implant guide in the early implant restoration of second molars. METHODS: The study included 35 patients with second molar loss randomly divided into two groups. Eighteen patients in the experimental group had an implant procedure using a computer-aided design/computer-assisted manufacturing (CAD/CAM) digital implant guide, and 17 patients in the control group had the procedure using a traditional film pressing implantation guide. Then, the surgical procedure was completed using the two different implant guides. At 3 months after surgery, four parameters including screw hole exit position, coronal deviation of the implant site, disease improvement rate, and clinical effects, which included marginal adaptation, anatomic form, marginal discoloration, postoperative sensitivity, surface roughness, and secondary caries of the upper prosthesis were compared between the two groups. RESULTS: The screw hole exit position in the experimental group was directed to the functional cusp of the opposite jaw, and there was a statistically significant difference between the two groups. There was no statistically significant difference in the rate of disease improvement and the clinical effect of the upper prosthesis between the two groups. There was no statistically significant difference in the bilateral coronal deviation and deviation direction of implants in the two groups. The bilateral coronal deviation of the experimental group was smaller than that of the control group. CONCLUSION: The digital implant guide can effectively reduce the deviation of the screw hole and the upper prosthesis in the restoration of the second molar. The prosthesis used in the experimental group had a good clinical outcome, which provides a theoretical basis for the restoration of the posterior molar.

Effect of perioperative cognitive behavioral interventions on pain, anxiety, and sleep quality in elderly patients after sinus floor elevation and immediate implantation: A randomized controlled trial.

BACKGROUND: Sinus floor elevation and immediate dental implantation are commonly performed to treat dentition defects in elderly patients. Targeted cognitive behavioral interventions (CBI) during the perioperative period can reduce pain and anxiety as well as improve sleep quality. This can lead to improvements in patient cooperation during follow-up treatment and enhance the overall efficacy of the surgery. OBJECTIVE: The study aimed to investigate the impact of a cognitive behavioral intervention method on perioperative pain, anxiety, and sleep quality in elderly patients undergoing sinus floor elevation and immediate dental implantation. METHODS: Forty patients who required the treatment at the Stomatology Clinic in our hospital between December 2018 and December 2022 were enrolled in this randomized controlled trial. The patients were randomly divided into two groups: a control group (n= 20), which received conventional treatment and care during the perioperative period, and an intervention group (n= 20), which received comprehensive behavioral intervention in addition to the conventional treatment and care during the perioperative period. The perioperative anxiety, pain, and sleep quality of the patients in both groups were evaluated. Anxiety was assessed using the Self-Rating Anxiety Scale (SAS), sleep quality was assessed using the Pittsburgh Sleep Quality Index (PSQI), and pain was measured using the visual analog scale (VAS). RESULTS: No statistically significant differences in SAS and PSQI were observed between the two groups at the initial visit; the values were significantly higher than those measured postoperatively. The SAS scores and PSQI of patients on days 0 and 7 post-surgery in the intervention group were significantly lower than those in the control group (P< 0.05). CONCLUSION: Perioperative cognitive behavioral intervention can effectively improve anxiety, postoperative pain and sleep quality in elderly patients who have undergone sinus floor elevation and immediate dental implantation, thereby reducing the incidence of complications.

The effect of motivational interviewing based on the transtheoretical model on oral cleaning behavior of patients with periodontitis who have undergone implant restoration.

BACKGROUND: Implant-restored patients with periodontitis have a higher risk of developing peri-implantitis, and helping them develop oral cleaning habits is significant. OBJECTIVE: To evaluate the effectiveness of motivational interviewing based on the transtheoretical model on the modification of oral cleaning behaviors of implant-restored patients with periodontitis. METHODS: Patients with periodontitis (n= 70) who would receive dental implant treatment were included. And they were randomly divided into two groups: experimental (n= 35) and control (n= 35). Control patients received routine oral hygiene education, and those in the experimental group received motivational interviewing based on the transtheoretical model. Oral cleaning behavior was compared between the two groups before and after intervention. In addition, periodontal health status was compared on the day of implant restoration and three months later. RESULTS: Compared to the control, the experimental group demonstrated significantly better oral hygiene behavior after intervention (P< 0.05). Three months after implant restoration, significantly better results were obtained by the experimental group in terms of mPLI and mSBI (P< 0.05). CONCLUSION: Motivational interviewing based on the transtheoretical model can effectively improve the oral cleaning behavior and periodontal health of implant-restored patients with periodontitis.

IL-36γ and IL-36Ra Reciprocally Regulate NSCLC Progression by Modulating GSH Homeostasis and Oxidative Stress-Induced Cell Death.

The balance between antioxidants and reactive oxygen species (ROS) critically regulates tumor initiation and progression. However, whether and how the tumor-favoring redox status is controlled by cytokine networks remain poorly defined. Here, it is shown that IL-36γ and IL-36Ra reciprocally regulate the progression of non-small cell lung cancer (NSCLC) by modulating glutathione metabolism and ROS resolution. Knockout, inhibition, or neutralization of IL-36γ significantly inhibits NSCLC progression and prolongs survival of the KrasLSL-G12D/+ Tp53fl/fl and KrasLSL-G12D/+ Lkb1fl/fl mice after tumor induction, whereas knockout of IL-36Ra exacerbates tumorigenesis in these NSCLC mouse models and accelerates death of mice. Mechanistically, IL-36γ directly upregulates an array of genes involved in glutathione homeostasis to reduce ROS and prevent oxidative stress-induced cell death, which is mitigated by IL-36Ra or IL-36γ neutralizing antibody. Consistently, IL-36γ staining is positively and negatively correlated with glutathione biosynthesis and ROS in human NSCLC tumor biopsies, respectively. These findings highlight essential roles of cytokine networks in redox for tumorigenesis and provide potential therapeutic strategy for NSCLC.

A single-cell survey of cellular hierarchy in acute myeloid leukemia.

BACKGROUND: Acute myeloid leukemia (AML) is a fatal hematopoietic malignancy and has a prognosis that varies with its genetic complexity. However, there has been no appropriate integrative analysis on the hierarchy of different AML subtypes. METHODS: Using Microwell-seq, a high-throughput single-cell mRNA sequencing platform, we analyzed the cellular hierarchy of bone marrow samples from 40 patients and 3 healthy donors. We also used single-cell single-molecule real-time (SMRT) sequencing to investigate the clonal heterogeneity of AML cells. RESULTS: From the integrative analysis of 191727 AML cells, we established a single-cell AML landscape and identified an AML progenitor cell cluster with novel AML markers. Patients with ribosomal protein high progenitor cells had a low remission rate. We deduced two types of AML with diverse clinical outcomes. We traced mitochondrial mutations in the AML landscape by combining Microwell-seq with SMRT sequencing. We propose the existence of a phenotypic "cancer attractor" that might help to define a common phenotype for AML progenitor cells. Finally, we explored the potential drug targets by making comparisons between the AML landscape and the Human Cell Landscape. CONCLUSIONS: We identified a key AML progenitor cell cluster. A high ribosomal protein gene level indicates the poor prognosis. We deduced two types of AML and explored the potential drug targets. Our results suggest the existence of a cancer attractor.

Long-read sequencing and de novo assembly of a Chinese genome.

Short-read sequencing has enabled the de novo assembly of several individual human genomes, but with inherent limitations in characterizing repeat elements. Here we sequence a Chinese individual HX1 by single-molecule real-time (SMRT) long-read sequencing, construct a physical map by NanoChannel arrays and generate a de novo assembly of 2.93 Gb (contig N50: 8.3 Mb, scaffold N50: 22.0 Mb, including 39.3 Mb N-bases), together with 206 Mb of alternative haplotypes. The assembly fully or partially fills 274 (28.4%) N-gaps in the reference genome GRCh38. Comparison to GRCh38 reveals 12.8 Mb of HX1-specific sequences, including 4.1 Mb that are not present in previously reported Asian genomes. Furthermore, long-read sequencing of the transcriptome reveals novel spliced genes that are not annotated in GENCODE and are missed by short-read RNA-Seq. Our results imply that improved characterization of genome functional variation may require the use of a range of genomic technologies on diverse human populations.

Activating mitochondrial function and haemoglobin expression with EH-201, an inducer of erythropoietin in neuronal cells, reverses memory impairment.

BACKGROUND AND PURPOSE: Memory impairment can be progressive in neurodegenerative diseases, and physiological ageing or brain injury, mitochondrial dysfunction and oxidative stress are critical components of these issues. An early clinical study has demonstrated cognitive improvement during erythropoietin treatment in patients with chronic renal failure. As erythropoietin cannot freely cross the blood-brain barrier, we tested EH-201 (2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucoside, also known as TSG), a low MW inducer of erythropoietin, for its therapeutic effects on memory impairment in models of neurodegenerative diseases, physiological ageing or brain injury. EXPERIMENTAL APPROACH: The effects of EH-201 were investigated in astrocytes and PC12 neuronal-like cells. In vivo, we used sleep-deprived (SD) mice as a stress model, amyloid-ß (Aß)-injected mice as a physiological ageing model and kainic acid (KA)-injected mice as a brain damage model to assess the therapeutic effects of EH-201. KEY RESULTS: EH-201 induced expression of erythropoietin, PPAR-γ coactivator 1α (PGC-1α) and haemoglobin in astrocytes and PC12 neuronal-like cells. In vivo, EH-201 treatment restored memory impairment, as assessed by the passive avoidance test, in SD, Aß and KA mouse models. In the hippocampus of mice given EH-201 in their diet, levels of erythropoietin, PGC-1α and haemoglobin were increased CONCLUSIONS AND IMPLICATIONS: The induction of endogenous erythropoietin in neuronal cells by inducers such as EH-201 might be a therapeutic strategy for memory impairment in neurodegenerative disease, physiological ageing or traumatic brain injury.

Genome-wide profiling of HPV integration in cervical cancer identifies clustered genomic hot spots and a potential microhomology-mediated integration mechanism.

Human papillomavirus (HPV) integration is a key genetic event in cervical carcinogenesis. By conducting whole-genome sequencing and high-throughput viral integration detection, we identified 3,667 HPV integration breakpoints in 26 cervical intraepithelial neoplasias, 104 cervical carcinomas and five cell lines. Beyond recalculating frequencies for the previously reported frequent integration sites POU5F1B (9.7%), FHIT (8.7%), KLF12 (7.8%), KLF5 (6.8%), LRP1B (5.8%) and LEPREL1 (4.9%), we discovered new hot spots HMGA2 (7.8%), DLG2 (4.9%) and SEMA3D (4.9%). Protein expression from FHIT and LRP1B was downregulated when HPV integrated in their introns. Protein expression from MYC and HMGA2 was elevated when HPV integrated into flanking regions. Moreover, microhomologous sequence between the human and HPV genomes was significantly enriched near integration breakpoints, indicating that fusion between viral and human DNA may have occurred by microhomology-mediated DNA repair pathways. Our data provide insights into HPV integration-driven cervical carcinogenesis.