Complete De Novo Assembly of Wolbachia Endosymbiont of Frankliniella intonsa.

As an endosymbiont, Wolbachia exerts significant effects on the host, including on reproduction, immunity, and metabolism. However, the study of Wolbachia in Thysanopteran insects, such as flower thrips Frankliniella intonsa, remains limited. Here, we assembled a gap-free looped genome assembly of Wolbachia strain wFI in a length of 1,463,884 bp (GC content 33.80%), using Nanopore long reads and Illumina short reads. The annotation of wFI identified a total of 1838 protein-coding genes (including 85 pseudogenes), 3 ribosomal RNAs (rRNAs), 35 transfer RNAs (tRNAs), and 1 transfer-messenger RNA (tmRNA). Beyond this basic description, we identified mobile genetic elements, such as prophage and insertion sequences (ISs), which make up 17% of the entire wFI genome, as well as genes involved in riboflavin and biotin synthesis and metabolism. This research lays the foundation for understanding the nutritional mutualism between Wolbachia and flower thrips. It also serves as a valuable resource for future studies delving into the intricate interactions between Wolbachia and its host.

The Chromosome-Level Genome Assembly of Bean Blossom Thrips (Megalurothrips usitatus) Reveals an Expansion of Protein Digestion-Related Genes in Adaption to High-Protein Host Plants.

Megalurothrips usitatus (Bagnall) is a destructive pest of legumes, such as cowpea. The biology, population dynamics and control strategies of this pest have been well studied. However, the lack of a high-quality reference genome for M. usitatus has hindered the understanding of key biological questions, such as the mechanism of adaptation to feed preferentially on high-protein host plants and the resistance to proteinase inhibitors (PIs). In this study, we generated a high-resolution chromosome-level reference genome assembly (247.82 Mb, 16 chromosomes) of M. usitatus by combining Oxford Nanopore Technologies (ONT) and Hi-C sequencing. The genome assembly showed higher proportions of GC and repeat content compared to other Thripinae species. Genome annotation revealed 18,624 protein-coding genes, including 4613 paralogs that were preferentially located in TE-rich regions. GO and KEGG enrichment analyses of the paralogs revealed significant enrichment in digestion-related genes. Genome-wide identification uncovered 506 putative digestion-related enzymes; of those, proteases, especially their subgroup serine proteases (SPs), are significantly enriched in paralogs. We hypothesized that the diversity and expansion of the digestion-related genes, especially SPs, could be driven by mobile elements (TEs), which promote the adaptive evolution of M. usitatus to high-protein host plants with high serine protease inhibitors (SPIs). The current study provides a valuable genomic resource for understanding the genetic variation among different pest species adapting to different plant hosts.

MicroRNA-145 influences the balance of Th1/Th2 via regulating RUNX3 in asthma patients.

AIM OF THE STUDY: To delineate the underlying mechanism of microRNA-145 modulate the balance of Th1/Th2 via targeting RUNX3 in asthma patients. MATERIALS AND METHODS: Peripheral blood samples were collected from asthma patients and healthy controls. CD4+ T cells were isolated and cultured. Using quantitative PCR detect, the level of microRNA-145 and RUNX3 mRNA level in the CD4+ T cells from asthma patients and healthy controls, meanwhile, western blot was used to detect the RUNX3 protein level. Th1 or Th2 related cytokines were measured by enzyme-linked immunosorbent assay. Dual-Luciferase Reporter Assay was performed to confirm the correlation between microRNA-145 and RUNX3. MicroRNA-145 mimic or inhibitor was transfected in the CD4+ T cells and the changes of RUNX3 level, Th1 or Th2 related cytokines and the percentage of Th1 and Th2 were observed after transfection. RESULTS: MicroRNA-145 level of CD4+ T cells was higher with a lower RUNX3 expression in asthma patients. There is negative correlation between microRNA-145 and RUNX3. Th2 hyperactivity and Th1 deficiency was detected in the CD4+ T cells of asthma patients. Dual-Luciferase Reporter Assay has shown that RUNX3 is a target of microRNA. Up-regulation or down-regulation of miR-145 level caused RUNX3 expression changes in CD4+ T cells and influence the related cytokines. Inhibition of microRNA-145 may reverse the imbalance of Th1/Th2 in asthma patients. CONCLUSION: MicroRNA-145 could regulate the balance of Th1/Th2 through targeting the RUNX3 in asthma patients. MicroRNA-145 and RUNX3 may be used as biomarkers or targets in the diagnosis or therapy of asthma.

[Effect of dust aerosol exposure on lung function and lung histopathology in rats].

OBJECTIVE: To investigate the effect of dust aerosol exposure on lung function and lung histopathology in rats. METHODS: According to random number table method, 120 Wistar male rats were divided into untreated control group, treated control group and experimental group, with 40 rats in each group. Experimental group were exposed to the wind tunnel simulation of sandstorm for 5 hours in every day; the untreated control group were put in the standard living environment next to the wind tunnel; the treated control group were exposed to the same wind tunnel simulation of sandstorm for 5 hours in every day, and the speed of wind was the same as the experimental group, but excluding dust. At different time points, the lung function and electron microscopy were performed in all rats. RESULTS: The level of Dynamic Compliance (Cdyn) ((0.227 ± 0.023), (0.198 ± 0.022) ml/cmH2O, 1 cmH2O=0.098 kPa) and forced vital capacity (FVC) ((6.24 ± 0.29), (5.59 ± 0.19) ml) were lower in the experimental group at 90 and 120 days, as compared to the untreated control group (Cdyn: (0.266 ± 0.014), (0.265 ± 0.018) ml/cmH2O; FVC: (7.15 ± 0.23), (7.17 ± 0.20) ml) and treated control group (Cdyn: (0.269 ± 0.015), (0.264 ± 0.019) ml/cmH2O; FVC: (7.14 ± 0.19), (7.15 ± 0.21) ml) (all P<0.05). At 120 days, The level of the forced expiratory flow after 50% of the FVC ((12.3 ± 2.2) ml/s) and peak expiratory flow ((25.79 ± 0.42) ml/s) were lower in the experimental group, as compared to the untreated control group ((15.9 ± 2.5), (27.99 ± 0.36) ml/s) and treated control group ((15.8 ± 2.1), (27.90 ± 0.38) ml/s) (all P<0.01). The FVC rate of 0.2 second in the experimental group was higher than that in the untreated control group and treated control group ( (85 ± 5)%, (73 ± 4)%, (73 ± 4)%, all P<0.05). The electron microscopy showed that the lung tissues had no obvious abnormalities at 30, 60, 90 and 120 days in untreated control group and treated control group. But in the experimental group, at 30 days, the endothelial cells of alveolar type I cells were swelled and the number of alveolar type II cells were increased; at 60 days, alveolar type II cells hyperplastic, basement membrane thinned and destructed; at 90 days, the number of alveolar type II cells decreased, Lamellar body evacuation; at 120 days, a lot of collagen fiber was formed in the alveolar septa. CONCLUSIONS: The strong sandstorm environmental exposure to a certain period of time can cause the decline of lung function and the damage of lung histopathology in rats. Exposure time was positively correlated with the damage of lung tissue.

Chromosome-level genome assembly of the flower thrips Frankliniella intonsa.

As an economically important insect pest, the flower thrips Frankliniella intonsa (Trybom) causes great damage to host plants by directly feeding and indirectly transmitting various pathogenic viruses. The lack of a well-assembled genomic resource has hindered our understanding of the genetic basis and evolution of F. intonsa. In this study, we used Oxford Nanopore Technology (ONT) long reads and High-through chromosome conformation capture (Hi-C) linked reads to construct a high-quality reference genome assembly of F. intonsa, with a total size of 225.5 Mb and a contig N50 of 3.37 Mb. By performing the Hi-C analysis, we anchored 91.68% of the contigs into 15 pseudochromosomes. Genomic annotation uncovered 17,581 protein-coding genes and identified 20.09% of the sequences as repeat elements. BUSCO analysis estimated over 98% of genome completeness. Our study is at the first time to report the chromosome-scale genome for the species of the genus Frankliniella. It provides a valuable genomic resource for further biological research and pest management of the thrips.

[Analysis of characteristics of the location of lower extremity deep vein thrombosis and the site of pulmonary embolism in hospitalized patients].

OBJECTIVE: To investigate the characteristics and relationship between the location of lower extremity deep vein thrombosis (DVT) and the site of pulmonary embolism in hospitalized patients. METHODS: The data of patients with lower extremity DVT diagnosed by ultrasound examination and pulmonary embolism diagnosed by CT pulmonary angiography from December 2017 to December 2021 were analyzed retrospectively. According to the location of lower extremity DVT, the patients were divided into mixed DVT, proximal DVT, and distal DVT which was further divided into anterior/posterior tibial vein or peroneal vein thrombosis and calf muscular venous thrombosis. Mixed DVT was referred to the presence of both proximal and distal DVT. According to the involved site of pulmonary artery, pulmonary embolism was divided into three types: main pulmonary artery, left or right pulmonary artery trunk embolism, lobar pulmonary artery embolism and segmental pulmonary artery embolism. The location of lower extremity DVT, the site of pulmonary embolism, the clinical manifestation (shortness of breath, chest tightness, chest pain, hemoptysis, cough, lower limb swelling, lower limb pain, syncope, fever) and risk factors (fracture/trauma, tumor, diabetes, hypertension, atrial fibrillation, infection, surgery, autoimmune diseases, paralysis, pregnancy) of venous thromboembolism (VTE), and the level of D-dimer were analyzed. RESULTS: A total of 209 patients were enrolled finally, including 127 patients with left lower extremity DVT (60.8%) and 82 with right lower extremity DVT (39.2%). Mixed DVT accounted for 39.2%, proximal DVT accounted for 17.3%, and distal DVT accounted for 43.5% (anterior/posterior tibial vein and peroneal vein thrombosis accounted for 14.8%, calf muscular venous thrombosis accounted for 28.7%). The incidences of main pulmonary artery embolism, left or right pulmonary artery trunk embolism in the mixed DVT and proximal DVT were significantly higher than those in the anterior/posterior tibial vein or peroneal vein thrombosis and calf muscular venous thrombosis [41.5% (34/82), 38.8% (14/36) vs. 16.2% (5/31), 10.0% (6/60)], with statistically significant differences (all P < 0.05). The incidences of pulmonary segmental artery embolism in the anterior/posterior tibial vein or peroneal vein thrombosis were higher than those in the mixed DVT and proximal DVT [41.9% (13/31) vs. 26.8% (22/82), 30.6% (11/36)], but the difference was not statistically significant (both P > 0.05). The incidences of pulmonary segmental artery embolism in the calf muscular venous thrombosis were significantly higher than those in the mixed DVT and the proximal DVT [66.7% (40/60) vs. 26.8% (22/82), 30.6% (11/36)], and the difference was statistically significant (both P < 0.05). The levels of D-dimer in patients with calf muscular venous thrombosis combined with main pulmonary artery embolism, left or right pulmonary artery trunk embolism were significantly higher than those in patients with calf muscular venous thrombosis combined pulmonary segmental artery embolism (mg/L: 6.08±3.12 vs. 3.66±2.66, P < 0.05). There were no significant differences in D-dimer levels in other patients with DVT combined with pulmonary embolism in different sites. In terms of the clinical manifestations of VTE, the incidences of lower limb swelling in the mixed DVT and proximal DVT were significantly higher than those in the anterior/posterior tibial vein or peroneal vein thrombosis and calf muscular venous thrombosis [54.9% (45/82), vs. 29.0% (9/31), 15.0% (9/60), both P < 0.05], the incidences of lower limb swelling in the proximal DVT were significantly higher than those in the calf muscular venous thrombosis [41.7% (15/63) vs. 15.0% (9/60), P < 0.05], there were no significant difference in the other clinical manifestations among the DVT groups. There was no significant difference in the incidence of VTE risk factors among the groups. CONCLUSIONS: The DVT of inpatients mostly occurred in the left lower limb, and the incidence of distal DVT was higher than that of proximal DVT. Mixed DVT and proximal DVT combined with pulmonary embolism mostly occurred in the main pulmonary artery, left or right pulmonary artery trunk, while distal DVT combined with pulmonary embolism mostly occurred in the pulmonary segmental artery. The levels of D-dimer in patients with lower extremity DVT combined with main pulmonary artery or left and right pulmonary artery trunk embolism were higher than those in patients with pulmonary lobe and segmental artery embolism. The incidence of lower extremity swelling in patients with mixed DVT and proximal DVT was higher than that in patients with distal DVT.

METTL3-Mediated N6-Methyladenosine Methylation Modifies Foxp3 mRNA Levels and Affects the Treg Cells Proportion in Peripheral Blood of Patients with Asthma.

OBJECTIVE: To investigate the regulatory effect and mechanism of methyltransferase-like protein 3 (METTL3)-mediated N6-methyladenosine methylation (m6A) on forkhead box protein 3 (Foxp3) levels and the proportion of regulatory T (Treg) cells in the peripheral blood of patients with asthma. METHODS: Flow cytometry and ELISA were used to detect the differences in the proportions of Treg cells and serum interleukins (ILs) 4 and 7, respectively, in the peripheral blood between healthy individuals and patients with different asthma conditions. Reverse transcription-quantitative PCR (RT-qPCR) and Western blotting were used to detect the mRNA and protein expression levels, respectively, of METTL3 and Foxp3 in CD4+ T cells in the peripheral blood samples of different groups. M6A blot and m6A coimmunoprecipitation-PCR were used to detect the global and Foxp3 mRNA m6A levels, respectively, in the peripheral blood CD4+ T cells. CD4+ T cells collected from the peripheral blood of patients with asthma were subjected to in vitro transfection to knockdown the METTL3 levels and observe changes in the Foxp3 mRNA, protein, m6A levels, and RNA stability. Flow cytometry and ELISA were used to detect the changes in the Treg cell proportion and IL-4 and IL-17 levels in the cell culture supernatant. RESULTS: Compared with the healthy individuals, the ratio of Treg cells to peripheral blood CD4+ T cells was significantly decreased and the Foxp3 mRNA and protein expression was downregulated in patients with asthma with disease progression. The Foxp3 mRNA and protein expression levels were positively correlated with the Treg cell proportion and negatively correlated with IL-17 expression. The global and Foxp3 mRNA m6A levels were increased in the peripheral blood CD4+ T cells of patients with asthma. METTL3 expression was significantly higher in the peripheral blood CD4+ T cells of patients with asthma compared with healthy individuals. After METTL3 knockdown, the Foxp3 mRNA m6A level was reduced, and the stability of Foxp3 mRNA and protein expression was increased. YTHDF2 could bind to the m6A site in 3'UTR of Foxp3 mRNA. Knockdown of YTHDF2 regulated the level and stability of Foxp3 mRNA. METTL3 knockdown reduced the ratio of Treg cells to CD4+ T cells and the IL-4 and IL-17 secretion levels from CD4+ T cells in the peripheral blood of patients with asthma. CONCLUSIONS: High METTL3 expression in the peripheral blood CD4+ T cells of patients with asthma increased the m6A level and reduced the stability of Foxp3 mRNA in a YTHDF2-dependent way, thereby reducing the expression of Foxp3 and the proportion of Treg cells.

Comparison of efficacy of stereotactic body radiotherapy and thoracoscopic surgery in the treatment of early-stage non-small cell lung cancer.

PURPOSE: To observe and compare the efficacy and safety between stereotactic body radiotherapy and thoracoscopic surgery in the treatment of early-stage non-small cell lung cancer (NSCLC). METHODS: The clinical data of 106 early-stage NSCLC patients admitted to the Thoracic Surgery Department of the hospital from February 2014 to February 2016 were retrospectively analyzed. Among these patients, 53 received stereotactic body radiotherapy (SBRT group), and 53 underwent video-assisted thoracoscopic surgery (VATS group). The clinical data of all patients were collected. The short-term response rate, Karnofsky performance status (KPS) score, changes in serum tumor marker levels before and after surgery and adverse reactions were compared between the two groups. Besides, all patients were followed up, and the overall survival (OS) and progression-free survival (PFS) were recorded. RESULTS: The levels of serum carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21-1) and neurone specific enolase (NSE) were decreased in both groups after treatment, and the differences were not statistically significant between the two groups. The patients tolerated well with SBRT, without evident myelosuppression or adverse hematological effects. In SBRT group, there were 7 cases of radioactive skin reaction, 2 cases of grade I radiation esophagitis and 4 cases of radiation pneumonitis (including 3 cases of grade I radiation pneumonitis and 1 case of grade II radiation pneumonitis). In VATS group, there were 3 cases of incision infection, 2 cases of pulmonary infection, 5 cases of pulmonary atelectasis, 1 case of pulmonary leakage and 1 case of deep vein thrombosis of lower extremity. The 3- year OS and PFS were 79.2% (42/53) and 67.9% (36/53) and 83.0% (44/53) and 77.4% (41/53) in SBRT group and VATS group, respectively. Kaplan-Meier survival showed no statistically significant differences in the OS and PFS between the two groups (log-rank). CONCLUSION: SBRT achieves better RR and DCR, similar OS and PFS to those of typical thoracoscopic surgery, and good patient tolerance in the treatment of early- stage NSCLC, which is a safe and effective treatment means.

Rare imaging findings of hypersensitivity pneumonitis: A case report.

BACKGROUND: Hypersensitivity pneumonitis (HP) is an immune-mediated syndrome caused by allergen inhalation. High-resolution computed tomography (HRCT) of HP may show diffuse ground-glass opacity, centrilobular ground-glass nodules, areas of air-trapping, thin-walled cysts, or fibrotic changes. CASE SUMMARY: A 47-year-old male patient went to the hospital complaining of cough and gradual aggravation of shortness of breath. HRCT of the lung showed that multiple nodules and ground-glass high-density shadows were present in both lungs. In addition, circular high-density shadows of various sizes were widely distributed in both lungs with relatively normal lung markings inside them. But other tests did not have a positive finding that can clarify the cause. Therefore, the patient underwent a lung biopsy. The pathological results showed that the lesions tended to be HP. After 4 mo of follow-up, the lesions in the patient's lungs were absorbed spontaneously, and the symptoms of cough and shortness of breath have disappeared. The review results suggested that the patient's disease was self-healing, which was consistent with the characteristics of HP. CONCLUSION: For some patients with HP, abnormal HRCT findings, such as the lesions in the lungs, can be absorbed on their own, which is an important clue in the diagnosis of the disease. Early diagnosis by lung biopsy is necessary when antigen exposure is unknown.