High diversity, close genetic relatedness, and favorable living conditions benefit species co-occurrence of gut microbiota in Brandt's vole.

Introduction: Revealing factors and mechanisms in determining species co-existence are crucial to community ecology, but studies using gut microbiota data are still lacking. Methods: Using gut microbiota data of 556 Brandt's voles from 37 treatments in eight experiments, we examined the relationship of species co-occurrence of gut microbiota in Brandt's voles (Lasiopodomys brandtii) with genetic distance (or genetic relatedness), community diversity, and several environmental variables. Results: We found that the species co-occurrence index (a larger index indicates a higher co-occurrence probability) of gut microbiota in Brandt's voles was negatively associated with the genetic distance between paired ASVs and the number of cohabitating voles in the experimental space (a larger number represents more crowding social stress), but positively with Shannon diversity index, grass diets (representing natural foods), and non-physical contact within an experimental space (representing less stress). Discussion: Our study demonstrated that high diversity, close genetic relatedness, and favorable living conditions would benefit species co-occurrence of gut microbiota in hosts. Our results provide novel insights into factors and mechanisms that shape the community structure and function of gut microbiota and highlight the significance of preserving the biodiversity of gut microbiota.

Discovery, Structure-Activity Relationship and In Vitro Anticancer Activity of Small-Molecule Inhibitors of the Protein-Protein Interactions between AF9/ENL and AF4 or DOT1L.

Chromosomal translocations involving the mixed lineage leukemia (MLL) gene cause 5-10% acute leukemias with poor clinical outcomes. Protein-protein interactions (PPI) between the most frequent MLL fusion partner proteins AF9/ENL and AF4 or histone methyltransferase DOT1L are drug targets for MLL-rearranged (MLL-r) leukemia. Several benzothiophene-carboxamide compounds were identified as novel inhibitors of these PPIs with IC50 values as low as 1.6 µM. Structure-activity relationship studies of 77 benzothiophene and related indole and benzofuran compounds show that a 4-piperidin-1-ylphenyl or 4-pyrrolidin-1-ylphenyl substituent is essential for the activity. The inhibitors suppressed expression of MLL target genes HoxA9, Meis1 and Myc, and selectively inhibited proliferation of MLL-r and other acute myeloid leukemia cells with EC50 values as low as 4.7 µM. These inhibitors are useful chemical probes for biological studies of AF9/ENL, as well as pharmacological leads for further drug development against MLL-r and other leukemias.

Livestock grazing is associated with the gut microbiota and antibiotic resistance genes in sympatric plateau pika (Ochotona curzoniae).

With the overuse of antibiotics in health care and animal husbandry, antibiotic resistance becomes a serious threat to public health. Antibiotic residues from veterinary medicine have increased the dissemination of antibiotic resistance genes (ARGs) by horizontal gene transfer globally, leading to the enrichment of ARGs in wildlife. Plateau pika (Ochotona curzoniae) is a small herbivore endemic to the Qinghai-Tibetan Plateau. Previous studies reveal that pika evolves a coprophagy behavior toward cohabitated yak, which makes the pika population a potential reservoir of ARGs. Yet, little is known about the resistome of pika under different grazing intensities. Here, we sampled the cecum content of pika from three different grazing intensity areas in the Qinghai-Tibetan Plateau to evaluate the effect of grazing on its gut microbiota and resistome. By using the 16S full-length amplicon and metagenomic sequencing, our study revealed that livestock grazing significantly altered the gut microbial community of plateau pika as compared to prohibited grazing areas. We found bacterial lineage Prevotellaceae, Lachnospirales, and RF39 increased in grazing areas. Analysis of the resistome revealed that pika from continuous grazing areas enriched a higher abundance of colistin (MCR) and streptogramin (vat) resistance genes. Moreover, we observed significant correlations between the gut microbial community, ARGs, and mobile genetic element profiles, hinting that pika gut microbiota was an important shaping force of the resistome. In future studies, the continuous monitoring of wildlife gut resistome and environmental antibiotic residues is imperative for a better understanding and for tackling the horizontal gene transfer of ARGs across the wildlife-livestock interface.

Health prevention intervention for chronic tissue fibrosis: Based on the specific expression of CCL19/21 + mast cell.

Chronic tissue fibrosis is a common pathological feature of connective tissue diseases and malignant tumors, and its prevention has been a major focus of relevant research.However, the details of the mechanism of action of tissue-colonizing immune cells in fibroblast migration are unclear. In this study, connective tissue disease tissue specimens and solid tumor specimens were selected to observe the relationship between mast cells and interstitial fibrosis and the expression characteristics of mast cells. Our findings suggest that the number of mast cells in the tissue correlates with the degree of pathological fibrosis and that mast cells specifically express the chemokines CCL19 and CCL21, especially CCL19. CCR7+ fibroblasts are highly expressed in mast cell clusters. The mast cell line HMC-1 regulates CD14+ monocyte-derived fibroblasts via CCL19. In disease tissue fibrosis, mast cell activation may increase the expression of chemokines, especially CCL19, in the tissue, thereby inducing a large number of CCR7-positive fibroblasts to migrate to specific tissues. This study lays a foundation for the mechanism of tissue fibrosis and provides evidence for the mechanism by which mast cells induce fibroblast migration.Through the experimental results of this paper, we can combine the induction factors of chronic tissue fibrosis and put forward targeted health prevention strategies.

Distal-continual colon interposition for esophageal reconstruction after esophagectomy: Two case reports.

Background: Colon interposition is a complex and time-consuming procedure requiring at least three or four digestive anastomoses. However, the long-term functional outcomes are promising, with an acceptable operative risk. Case presentation: Herein, two cases of esophageal carcinoma that received esophagus reconstruction using the distal continual colon interposition technique have been described. The transverse colon was lifted to the thoracic cavity for the end-to-side anastomosis with the esophagus, and a closure device was used to close the colon instead of severing and isolating the distal end. The duration of the operation was 140 and 150 min, respectively. The blood supply of the colon was maintained during the intervention. The tension-free anastomosis was performed without severe complications, and oral food intake was resumed on postoperative day 6. Neither anastomotic stenosis, antiacid or heartburn, dysphagia, or emptying obstacles nor complaints of diarrhea, bloating, or malodor were reported during the follow-up period. Conclusions: The modified distal-continual colon interposition technique may have the advantages of a short operation time and potential prevention of serious complications caused by the torsion of mesocolon vessels.

Effects of Group Size on Behavior, Reproduction, and mRNA Expression in Brains of Brandt's Voles.

For social animals, a moderate group size is greatly important to maintain their reproductive success. However, the underlying neurobiological mechanism of group size on behavior and reproduction has rarely been investigated. In this study, we examined the effects of group size (1, 2, 4 pairs of adult male and female voles raised per cage) on behavior and reproduction. Meanwhile, the mRNA expression of stress and reproduction response-related genes in male brains was detected. We found that Brandt's voles (Lasiopodomys brandtii) in the large-sized group fight more severely than those in the small-sized group. Meanwhile, male voles were more anxious than females. The average number of embryos and litters per female in the medium-sized group was significantly higher than that of large-sized group. In male voles, stress- or reproduction-response mRNA expressions were more related to final group size or final density due to death caused by fighting. Our results indicated that a moderate group size was beneficial to the reproductive output of Brandt's voles. Our study highlights the combined effects of stress- or reproduction-related gene expression or behavior in regulating the fitness of voles with different group sizes.

Impacts of Dietary Protein and Niacin Deficiency on Reproduction Performance, Body Growth, and Gut Microbiota of Female Hamsters (Tscherskia triton) and Their Offspring.

Food resources are vital for animals to survive, and gut microbiota play an essential role in transferring nutritional materials into functional metabolites for hosts. Although the fact that diet affects host microbiota is well known, its impacts on offspring remain unclear. In this study, we assessed the effects of low-protein and niacin-deficient diets on reproduction performance, body growth, and gut microbiota of greater long-tailed hamsters (Tscherskia triton) under laboratory conditions. We found that maternal low-protein diet (not niacin deficiency) had a significant negative effect on reproduction performance of female hamsters (longer mating latency with males and smaller litter size) and body growth (lower body weight) of both female hamsters and their offspring. Both protein- and niacin-deficient diets showed significant maternal effects on the microbial community in the offspring. A maternal low-protein diet (not niacin deficiency) significantly reduced the abundance of major bacterial taxa producing short-chain fatty acids, increased the abundance of probiotic taxa, and altered microbial function in the offspring. The negative effects of maternal nutritional deficiency on gut microbiota are more pronounced in the protein group than the niacin group and in offspring more than in female hamsters. Our results suggest that a low-protein diet could alter gut microbiota in animals, which may result in negative impacts on their fitness. It is necessary to conduct further analysis to reveal the roles of nutrition, as well as its interaction with gut microbes, in affecting fitness of greater long-tailed hamsters under field conditions. IMPORTANCE Gut microbes are known to be essential for hosts to digest food and absorb nutrients. Currently, it is still unclear how maternal nutrient deficiency affects the fitness of animals by its effect on gut microbes. Here, we evaluated the effects of protein- and niacin-deficient diets on mating behavior, reproduction, body growth, and gut microbiota of both mothers and offspring of the greater long-tailed hamster (Tscherskia triton) under laboratory conditions. We found that a low-protein diet significantly reduced maternal reproduction performance and body growth of both mothers and their offspring. Both protein and niacin deficiencies showed significant maternal effects on the microbial community of the offspring. Our results hint that nutritional deficiency may be a potential factor in causing the observed sustained population decline of the greater long-tailed hamsters due to intensified monoculture in the North China Plain, and this needs further field investigation.

The Survival and Prognosis Characteristics of Primary Esophageal Small-Cell Carcinoma.

Objective: To comprehensively explore the survival characteristics of primary esophageal small-cell carcinoma (PSCCE) and identify the main factors affecting the prognosis. Methods: The clinical and follow-up data of PSCCE patients admitted to the Fourth Hospital of Hebei Medical University from 2006 to 2010 were retrospectively analyzed. The primary endpoint was five-year survival. Survival curves were drawn using the Kaplan-Meier method, and log-rank test was used to compare the differences in survival rates among the groups. Cox regression models were used to analyze prognostic factors. Results: A total of 119 eligible patients were retrieved. Median survival was 27 months (3-100 months). Changes in overall survival (OS) in PSCCE patients were associated with TNM stage (P = 0.007), T stage (P = 0.049), and lymph node metastasis (P = 0.004). When TNM was in stage I-IIb, lymph node metastasis (P = 0.003) or combined adjuvant therapy (P = 0.004) was an independent factor affecting OS. Survival analysis showed that TNM staging had no predictive value for 5-year survival time or disease-free survival (DFS) of PSCCE (P > 0.05). Conclusion: TNM stage, T stage, and lymph node metastasis were related to the survival of patients. Negative lymph node metastasis and treatment are independent prognostic factors in PSCCE TNM stage I-IIb patients.

Abnormal TACC3 Expression is an Independent Prognostic Biomarker in Lung Carcinoma.

OBJECTIVE: Abnormal expression of transforming acidic coiled-coil protein 3 (TACC3) has been reported in many types of human malignancies. However, the expression of TACC3 and its clinical significance have not been well characterized in lung carcinoma (LUAD). The aim of this study was to investigate possible associations between tumor expression of TACC3 and the clinicopathological characteristics and prognosis of LUAD patients. METHODS: The expression of TACC3 in LUAD patients was determined using the Oncomine, Gene Expression Profiling Interactive Analysis (GEPIA), UALCAN, and Cancer Genome Atlas (TCGA) databases. The expression of TACC3 in LUAD tissues was also determined by qRT-PCR. RESULTS: TACC3 was found to be significantly overexpressed in LUAD tumors compared with non-tumor tissue in the above public databases. Receiver operating characteristic (ROC) curve analysis indicated that TACC3 could have diagnostic value in LUAD patients. Kaplan-Meier analysis further indicated that high TACC3 expression in tumors was significantly associated with worse overall survival (OS) in LUAD patients. In addition, univariate and multivariate Cox regression analyses showed that high TACC3 expression was an independent factor for worse OS in LUAD patients. Finally, based on gene set enrichment analysis (GSEA 3.0), we identified several potential pathways related to TACC3 that were enriched in the high TACC3 expression phenotype. CONCLUSIONS: The present study provides evidence that TACC3 expression is upregulated in LUAD and may be an independent risk factor for worse prognosis in these patients.

Posttreatment Recurrence and Death Patterns in Patients with Advanced Esophageal Cancer.

Objective: To investigate into the clinical factors associated with posttreatment recurrence and death patterns in patients with advanced esophageal cancer. Methods: Clinical information of patients with recurrence/metastasis and death after radical resection of esophageal cancer at our hospital between January 1, 2005, and December 31, 2015, were retrospectively collected and followed up. Postoperative recurrence-free survival time, postrelapse survival time, and overall survival time were compared among the metabolic-associated, organ failure-associated, and anastomotic recurrence-associated mortality groups. Results: Five hundred and ninety-five qualified patients were retrieved, including 456 males and 139 females, with an average age of 58 ± 7.56 years. There were 57 cases of TNM-1 stage, 131 cases of TNM-2 stage, 365 cases of TNM-3 stage, and 42 cases of TNM-4 stage. There were 547 cases of squamous cell cancer and 48 cases of nonsquamous cell cancer. There were significant differences in age (p < 0.01), tumor location (p < 0.01), and lymph node metastasis (p = 0.04), recurrence type (p < 0.01) by one-way ANOVA, and recurrence-free survival (p = 0.02) and postrecurrence survival (p < 0.01) by Kaplan-Meier survival curve analysis among the three main death causes. Conclusions: Age, tumor location, and lymph node metastasis were significantly different among metabolic-associated, organ failure-associated, and anastomotic recurrence-associated mortality of recurrent EC patients.

Aza-Quasi-Favorskii Reaction: Construction of Highly Substituted Aziridines through a Concerted Multibond Rearrangement Process.

A new molecular rearrangement, the aza-Quasi-Favorskii rearrangement, has been developed for the construction of highly substituted aziridines. Electron-deficient O-sulfonyl oximes react readily with α,α-disubstituted acetophenone-derived enolates to furnish highly substituted aziridines via this unprecedented domino process. In-depth computational studies reveal an asynchronous yet concerted nitrenoid-type rearrangement pathway.

Folate Receptor 4-Expressing T cell Is Associated with Disease-Free Survival in Patients with Esophageal Squamous Cell Carcinoma.

Background: Folic acid receptor 4 (FR4) significantly downregulates the expression of regular T cells (Treg) and improves the effect of chemotherapy and PD-1/PD-L1 inhibitors. However, the FR4 expression in squamous cell carcinoma (ESCC) remains unclear. Methods: Patients with primary ESCC who visited our hospital between 1st February 2012 and 30th September 2016 were enrolled in this study. FR4 expressions in ESCC patients were detected by immunohistochemistry staining, and the association with clinical characteristics and the overall survival (OS) or disease-free survival (DFS) was analyzed. Results: One hundred and forty-eight qualified cases of ESCC patients were retrieved, including 34 females. Ninety-four cases had lymph node metastasis (63.51%), 104 patients received adjuvant therapy (70.27%), and the rate of FR4 positive was 67.57% (100/148). Among FR4 positive patients, 75 cases received adjuvant therapy, and patients who received chemotherapy were significantly better than that of patients who did not receive chemotherapy. In patients with FR4 negative expression, 48 cases received adjuvant therapy, which was significantly worse than that of patients who did not receive chemotherapy. Conclusions: Postoperative adjuvant chemotherapy prolonged the survival in FR4 positive ESCC patients, whereas adjuvant therapy in patients with FR4 negative needs to be further improved.

Long-Term Effects of Climate Variability on Seed Rain Dynamics of Four Fagaceae Sympatric Species in Qinling Mountains, China.

Seed rain, as the beginning of species dispersal, is a key process for forest structure and regeneration. In this study, the seed rain of four Fagaceae sympatric plant species (Castaneamollissima, Quercus aliena, Quercus variabilis, and Quercus serrata) in the Qinling Mountains were monitored for ten consecutive years, and the responses of seed rain dynamics of the four species to major climatic factors (temperature and precipitation) were analyzed. We found there were significant differences in the seed rain dynamics between C. mollissima of Castanea and the other three species of Quercus in the initial period and end period and the duration of the whole seed rain process among the 10 years. This could indicate to some extent that there was no concentrated flowering and fruiting among different plants of different genera, and they could well avoid fierce competition for similar resources and coexist in the same region. This may also be a reproductive strategy for plants. Seed rain dynamics of different plant species had different sensitivities to climate factors (temperature and precipitation), which indicated that mainly because of their different responses to climate factors, they could well avoid fierce competition for similar climate resources. In addition, the differences in seed rain dropping dynamics could reduce consumption in large numbers by seed predators, thereby promoting their own dispersal and regeneration. All of the above contribute to their better coexistence in the same domain.

Mechanically evoked defensive attack is controlled by GABAergic neurons in the anterior hypothalamic nucleus.

Innate defensive behaviors triggered by environmental threats are important for animal survival. Among these behaviors, defensive attack toward threatening stimuli (for example, predators) is often the last line of defense. How the brain regulates defensive attack remains poorly understood. Here we show that noxious mechanical force in an inescapable context is a key stimulus for triggering defensive attack in laboratory mice. Mechanically evoked defensive attacks were abrogated by photoinhibition of vGAT+ neurons in the anterior hypothalamic nucleus (AHN). The vGAT+ AHN neurons encoded the intensity of mechanical force and were innervated by brain areas relevant to pain and attack. Activation of these neurons triggered biting attacks toward a predator while suppressing ongoing behaviors. The projection from vGAT+ AHN neurons to the periaqueductal gray might be one AHN pathway participating in mechanically evoked defensive attack. Together, these data reveal that vGAT+ AHN neurons encode noxious mechanical stimuli and regulate defensive attack in mice.

Apatinib Combined with Paclitaxel and Cisplatin Neoadjuvant Chemotherapy for Locally Advanced Esophageal Squamous Cell Carcinoma.

Background: We conducted a trial to evaluate the efficacy and safety of apatinib, a tyrosine inhibitor against vascular endothelial growth factor receptor 2, combined with neoadjuvant chemotherapy in patients with locally advanced esophageal squamous cell carcinoma (ESCC). Materials and Methods: One hundred twenty six patients were randomized into two cycles of paclitaxel and cisplatin (TP) (n = 61) or combined with apatinib (Apa+TP) (n = 65), followed by surgery. The primary endpoint was objective response rate (ORR). Secondary endpoints included pathological complete response (pCR), safety, R0 resection rate, and operative complication rates. Results: Compared with TP chemotherapy alone, adding apatinib to neoadjuvant treatment significantly increased ORR (Apa+TP: 80.0% vs. TP: 54.1%, respectively; p = 0.004). Apa+TP achieved higher pCR rate compared with TP alone (15.4% vs. 4.92%, respectively; p = 0.101). Similar incidences of toxic effects were found between those two groups. No grade 3 or 4 adverse events (AEs) were observed. Meanwhile, apatinib-related AEs, including hypertension, proteinuria, and hand-and-foot syndrome, were mild. The R0 resection rate was 100% in both groups. No significant differences in operation time, intraoperative bleeding, and postoperative complications were observed, and no serious complications occurred. Conclusions: Adding apatinib to TP neoadjuvant chemotherapy significantly increased ORR, suggesting an advantage of anti-angiogenesis in ESCC. Clinical Trials.gov ID: ChiCTR-TRC-1800017662.

Population variation alters aggression-associated oxytocin and vasopressin expressions in brains of Brandt's voles in field conditions.

Density-dependent change in aggressive behavior contributes to the population regulation of many small rodents, but the underlying neurological mechanisms have not been examined in field conditions. We hypothesized that crowding stress and aggression-associated oxytocin (OT) and arginine vasopressin (AVP) in specific regions of the brain may be closely related to aggressive behaviors and population changes of small rodents. We analyzed the association of OT and AVP expression, aggressive behavior, and population density of Brandt's voles in 24 large semi-natural enclosures (0.48 ha each) in Inner Mongolia grassland. We tested the effects of population density on the OT/AVP system and aggressive behavior by experimentally manipulating populations of Brandt's voles in the grassland enclosures. High density was positively and significantly associated with more aggressive behavior, and increased expression of mRNA and protein of AVP and its receptor, but decreased expression of mRNA and protein of OT and its receptor in specific brain regions of the voles. Our study suggests that changes in OT/AVP expression are likely a result of the increased psychosocial stress that these voles experience during overcrowding, and thus the OT/AVP system can be used as indicators of density-dependent stressors in Brandt's voles.

Timing outweighs magnitude of rainfall in shaping population dynamics of a small mammal species in steppe grassland.

Climate change-induced shifts in species phenology differ widely across trophic levels, which may lead to consumer-resource mismatches with cascading population and ecosystem consequences. Here, we examined the effects of different rainfall patterns (i.e., timing and amount) on the phenological asynchrony of population of a generalist herbivore and their food sources in semiarid steppe grassland in Inner Mongolia. We conducted a 10-y (2010 to 2019) rainfall manipulation experiment in 12 0.48-ha field enclosures and found that moderate rainfall increases during the early rather than late growing season advanced the timing of peak reproduction and drove marked increases in population size through increasing the biomass of preferred plant species. By contrast, greatly increased rainfall produced no further increases in vole population growth due to the potential negative effect of the flooding of burrows. The increases in vole population size were more coupled with increased reproduction of overwintered voles and increased body mass of young-of-year than with better survival. Our results provide experimental evidence for the fitness consequences of phenological mismatches at the population level and highlight the importance of rainfall timing on the population dynamics of small herbivores in the steppe grassland environment.

Synthesis, structure-activity relationship and antiviral activity of indole-containing inhibitors of Flavivirus NS2B-NS3 protease.

Zika virus belongs to the Flavivirus family of RNA viruses, which include other important human pathogens such as dengue and West Nile virus. There are no approved antiviral drugs for these viruses. The highly conserved NS2B-NS3 protease of Flavivirus is essential for the replication of these viruses and it is therefore a drug target. Compound screen followed by medicinal chemistry optimization yielded a novel series of 2,6-disubstituted indole compounds that are potent inhibitors of Zika virus protease (ZVpro) with IC50 values as low as 320 nM. The structure-activity relationships of these and related compounds are discussed. Enzyme kinetics studies show the inhibitor 66 most likely exhibited a non-competitive mode of inhibition. In addition, this series of ZVpro inhibitors also inhibit the NS2B-NS3 protease of dengue and West Nile virus with reduced potencies. The most potent compounds 66 and 67 strongly inhibited Zika virus replication in cells with EC68 values of 1-3 µM. These compounds are novel pharmacological leads for further drug development targeting Zika virus.

Small-molecule inhibitor of AF9/ENL-DOT1L/AF4/AFF4 interactions suppresses malignant gene expression and tumor growth.

Chromosome translocations involving mixed lineage leukemia (MLL) gene cause acute leukemia with a poor prognosis. MLL is frequently fused with transcription cofactors AF4 (~35%), AF9 (25%) or its paralog ENL (10%). The AHD domain of AF9/ENL binds to AF4, its paralog AFF4, or histone-H3 lysine-79 (H3K79) methyltransferase DOT1L. Formation of AF9/ENL/AF4/AFF4-containing super elongation complexes (SEC) and the catalytic activity of DOT1L are essential for MLL-rearranged leukemia. Protein-protein interactions (PPI) between AF9/ENL and DOT1L/AF4/AFF4 are therefore a potential drug target. Methods: Compound screening followed by medicinal chemistry was used to find inhibitors of such PPIs, which were examined for their biological activities against MLL-rearranged leukemia and other cancer cells. Results: Compound-1 was identified to be a novel small-molecule inhibitor of the AF9/ENL-DOT1L/AF4/AFF4 interaction with IC50s of 0.9-3.5 µM. Pharmacological inhibition of the PPIs significantly reduced SEC and DOT1L-mediated H3K79 methylation in the leukemia cells. Gene profiling shows compound-1 significantly suppressed the gene signatures related to onco-MLL, DOT1L, HoxA9 and Myc. It selectively inhibited proliferation of onco-MLL- or Myc-driven cancer cells and induced cell differentiation and apoptosis. Compound-1 exhibited strong antitumor activity in a mouse model of MLL-rearranged leukemia. Conclusions: The AF9/ENL-DOT1L/AF4/AFF4 interactions are validated to be an anticancer target and compound-1 is a useful in vivo probe for biological studies as well as a pharmacological lead for further drug development.

Poor Prognosis of Pulmonary Adenosquamous Carcinoma with NRAS and HRAS Double Mutation.

RAS mutations constitute one of the major tumorigenic mechanisms and are detected in approximately 20% of lung cancers. The most frequent mutated and well-studied RAS isoform is KRAS, which is associated with an overall poor prognosis in non-small-cell lung cancer (NSCLC). However, the clinical significances of NRAS and HRAS in NSCLC are rarely reported. Here, we present a 58-year-old male smoker who was diagnosed with stage IV lung adenosquamous carcinoma. A rare NRAS and HRAS double mutation was detected in the primary tumor and lymph node samples using next-generation sequencing (NGS). The patient showed rapid disease progression and passed away due to respiratory failure after 15 days of osimertinib in combination with cisplatin. To the best of our knowledge, this is the first report associating NRAS and HRAS double mutation in the poor prognosis of NSCLC.