Regulatory role and translational potential of CCL11 in liver fibrosis.

BACKGROUND AND AIMS: Myofibroblasts are considered the major effector cell type of liver fibrosis and primarily derived from hepatic stellate cells (HSCs). In the present study, we investigated the contribution of C-C motif chemokine (CCL11) to HSC-myofibroblast trans -differentiation and its implication in liver fibrosis. APPROACH AND RESULTS: We report that CCL11 levels were elevated in HSCs, but not in hepatocytes or Kupffer cells, isolated from mice with liver fibrosis compared with the control mice. CCL11 levels were also up-regulated by 2 pro-fibrogenic growth factors TGF-ß and platelet derived growth factor in cultured HSCs. Mechanistically, zinc finger factor 281 bound to the CCL11 promoter and mediated CCL11 trans -activation in HSCs. Depletion of CCL11 attenuated whereas treatment with recombinant CCL11 promoted HSC activation. Further, global CCL11 deletion ( CCL11-/- ) or HSC/myofibroblast-specific CCL11 knockdown mitigated fibrogenesis in mice. RNA-sequencing revealed that CCL11 might regulate HSC activation by stimulating the transcription of Jagged 1. Reconstitution of Jagged 1 restored the fibrogenic response in CCL11-/- mice. Finally, several targeting strategies that aimed at blockading CCL11 signaling, either by administration of an antagonist to its receptor C-C motif chemokine receptor 3 or neutralizing antibodies against CCL11/C-C motif chemokine receptor 3, ameliorated liver fibrosis in mice. CONCLUSIONS: Our data unveil a previously unrecognized role for CCL11 in liver fibrosis and provide proof-of-concept evidence that targeting CCL11 can be considered as an effective therapeutic approach.

The lncRNA TRG-AS1 promotes the growth of colorectal cancer cells through the regulation of P2RY10/GNA13.

BACKGROUND: The lncRNA TRG-AS1 and its co-expressed gene P2RY10 are important for colorectal cancer (CRC) occurrence and development. The purpose of our research was to explore the roles of TRG-AS1 and P2RY10 in CRC progression. METHODS: The abundance of TRG-AS1 and P2RY10 in CRC cell lines (HT-29 and LoVo) and normal colon cells FHC was determined and difference between CRC cells and normal cells was compared. LoVo cells were transfected with si-TRG-AS1 and si-P2RY10 constructs. Subsequently, the viability, colony formation, and migration of the transfected cells were analyzed using cell counting kit-8, clonogenicity, and scratch-wound/Transwell® assays, respectively. Cells overexpressing GNA13 were used to further explore the relationship between TRG-AS1 and P2RY10 along with their downstream functions. Finally, nude mice were injected with different transfected cell types to observe tumor formation in vivo. RESULTS: TRG-AS1 and P2RY10 were significantly upregulated in HT-29 and LoVo compared to FHC cells. TRG-AS1 knockdown and P2RY10 silencing suppressed the viability, colony formation, and migration of LoVo cells. TRG-AS1 knockdown downregulated the expression of P2RY10, GNA12, and GNA13, while P2RY10 silencing downregulated the expression of TRG-AS1, GNA12, and GNA13. Additionally, GNA13 overexpression reversed the cell growth and gene expression changes in LoVo cells induced by TRG-AS1 knockdown or P2RY10 silencing. In vivo experiments revealed that CRC tumor growth was suppressed by TRG-AS1 knockdown and P2RY10 silencing. CONCLUSIONS: TRG-AS1 knockdown repressed the growth of HT-29 and LoVo by regulating P2RY10 and GNA13 expression.

Withaferin A suppressed hepatocellular carcinoma progression through inducing IGF2BP3/FOXO1/JAK2/STAT3 pathway-mediated ROS production.

OBJECTIVE: This study aimed to investigate the underlying molecular mechanisms of Withaferin A (WA) in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: The gene and protein expression were analyzed using RT-qPCR and western blot, respectively. The proliferation of HCC cells was evaluated by CCK-8 assays. The migrative ability of HCC cells was measured by transwell assays. RESULTS: We revealed that WA suppressed the proliferation and migration of HCC cells and inhibited IGF2BP3 (insulin like growth factor 2 mRNA binding protein 3) expression. IGF2BP3 abundance reversed the reactive oxygen species (ROS) accumulation and suppression of HCC cell proliferation and migration induced by WA. Besides, IGF2BP3 suppressed ROS production to promote the growth and migration of HCC cells. Furthermore, we found that IGF2BP3 exerted its tumor-promotive and ROS-suppressive effect on HCC cells by regulating the expression of FOXO1 (forkhead box O1). In addition, IGF2BP3-stimulated activation of JAK2 (Janus kinase 2)/STAT3 (signal transducer and activator of transcription 3) phosphorylation effectively decreased the transcription of FOXO1. FOXO1 abundance decreased the phosphorylation of JAK2 and STAT3 by increasing ROS level, forming a feedback loop for the inhibition of JAK2/STAT3 signaling activated by IGF2BP3. CONCLUSIONS: WA-induced ROS inhibited HCC cell growth and migration through the inhibition of IGF2BP3 to deactivate JAK2/STAT3 signaling, resulting in increased FOXO1 expression to further stimulate ROS production and inhibit JAK2/STAT3 signaling.

Comparative Analyses of Reproductive Caste Types Reveal Vitellogenin Genes Involved in Queen Fertility in Solenopsis invicta.

The red imported fire ant (Solenopsis invicta Buren) is a social pest species with a robust reproductive ability that causes extensive damage. Identification of the genes involved in queen fertility is critical in order to better understand the reproductive biology and screening for the potential molecular targets in S. invicta. Here, we used the mRNA deep sequencing (RNA-seq) approach to identify differentially expressed genes (DEGs) in the transcriptomes of three reproductive caste types of S. invicta, including queen (QA) and winged female (FA) and male (MA) ants. The genes that were specific to and highly expressed in the queens were then screened, and the Vg2 and Vg3 genes were chosen as targets to explore their functions in oogenesis and fertility. A minimum of 6.08 giga bases (Gb) of clean reads was obtained from all samples, with a mapping rate > 89.78%. There were 7524, 7133, and 977 DEGs identified in the MA vs. QA, MA vs. FA, and FA vs. QA comparisons, respectively. qRT-PCR was used to validate 10 randomly selected DEGs, including vitellogenin 2 (Vg2) and 3 (Vg3), and their expression patterns were mostly consistent with the RNA-seq data. The S. invicta Vgs included conserved domains and motifs that are commonly found in most insect Vgs. SiVg2 and SiVg3 were highly expressed in queens and winged females and were most highly expressed in the thorax, followed by the fat body, head, and epidermis. Evaluation based on a loss-of-function-based knockdown analysis showed that the downregulation of either or both of these genes resulted in smaller ovaries, less oogenesis, and less egg production. The results of transcriptional sequencing provide a foundation for clarifying the regulators of queen fertility in S. invicta. The functions of SiVg2 and SiVg3 as regulators of oogenesis highlight their importance in queen fecundity and their potential as targets of reproductive disruption in S. invicta control.

Application of endoscopic ultrasound-guided hepaticogastrostomy combined with antegrade stenting in patients with malignant biliary obstruction after failed ERCP.

BACKGROUND: This study was aimed at comparing the safety and effectiveness of endoscopic ultrasound-guided hepaticogastrostomy (EUS-HGS) and EUS-HGS combined with antegrade stenting (EUS-HGAS) in patients with malignant biliary obstruction (MBO) after failed endoscopic retrograde cholangiopancreatography (ERCP). METHODS: Patients diagnosed with MBO and receiving EUS-HGS or EUS-HGAS from September 2015 to October 2020 were enrolled in this study. Clinical success, complications, reintervention rate, post-operative hospital stay, time to stent dysfunction, and patient death were compared. RESULTS: A total of 45 patients (21 in the EUS-HGAS group and 24 in the EUS-HGS group) were enrolled in this study. In the EUS-HGAS group, 21 patients all achieved clinical success (100%); in the EUS-HGS group, 24 patients also achieved technical success (100%) (P > 0.05). The differences between pre- and post-operative TB and ALT and AST levels were greater in the single-step EUS-HGAS group (P < 0.05). The incidence of complications was 2 of 21 (9.5%) in the EUS-HGAS group and 5 of 24 (20.8%) in the EUS-HGS group (P > 0.05). The reintervention rate was 0 in the EUS-HGAS group and 1 (4.2%) in the EUS-HGS group (P > 0.05). Time to stent dysfunction or patient death was longer in the EUS-HGAS group (P < 0.05). The post-operative hospital stay was longer and the total cost was higher in the EUS-HGAS group. CONCLUSION: EUS-HGAS was superior to EUS-HGS in terms of biliary drainage effectiveness and time to stent dysfunction or patient death in patients with MBO after failed ERCP. Furthermore, two-step EUS-HGAS may be safer in some patients.

Circulating exosome-derived bona fide long non-coding RNAs predicting the occurrence and metastasis of hepatocellular carcinoma.

Although the diagnosis and therapy approach developed, techniques for the early diagnosis of HCC remain insufficient which results in poor prognosis of patients. The traditional biomarker AFP, however, has been proved with low specificity. Circulating exosomal ncRNAs revealed different profiles reflecting the characteristics of tumour. In this study, we mainly focused on circulating exosomal ncRNAs which might be the fingerprint for HCC, especially for the diagnosis or metastasis prediction. A high throughput lncRNA microarray in exosomes extracted from cell-free plasma was applied. The risk score analysis was employed to screen the potential exosome-derived lncRNAs in two independent sets based on different clinical parameters in 200 paired HCC patients. After a multi-stage validation, we finally revealed three lncRNAs, ENSG00000248932.1, ENST00000440688.1 and ENST00000457302.2, increased in HCC comparing with the both chronic hepatitis (CH) patients and cancer-free controls. ROC curve revealed a higher sensitivity and specificity in predicting the occurrence of HCC from cancer-free controls and CH patients with the area under curve (AUC) of 0.905 and 0.879 by combining AFP. The three lncRNA panel combined with AFP also indicted a fingerprint function in predicting the metastasis of HCC with the AUC of 0.870. In conclusion, ENSG00000248932.1, ENST00000440688.1 and ENST00000457302.2 might be the potential biomarker for the tumorigenesis prediction from CH patients or healthy controls and may also be applied for dynamic monitoring the metastasis of HCC.

LncRNA SNHG14 potentiates pancreatic cancer progression via modulation of annexin A2 expression by acting as a competing endogenous RNA for miR-613.

This study aimed to determine long non-coding RNA (lncRNA) small nucleolar RNA host gene 14 (SNHG14) expression in pancreatic cancer and to explore the potential molecular actions of SNHG14 in mediating pancreatic cancer progression. Gene expression was detected by quantitative real-time PCR. Cell proliferation, growth and invasion were detected by respective CCK-8, colony formation, and transwell invasion assays. Protein levels were measured by Western blotting. Cell apoptosis and caspase-3 activity were detected by flow cytometry and caspase-3 activity assay. The link between miR-613 and its targets was evaluated by luciferase reporter assay. In vivo tumour growth was evaluated using a xenograft model of nude mice. SNHG14 expression was up-regulated in cancerous tissues from pancreatic cancer patients. High expression of SNHG14 was associated with poor tumour differentiation, advanced TNM stage and nodal metastasis. SNHG14 overexpression enhanced cell proliferative, growth and invasive abilities, and suppressed apoptotic rates and caspase-3 activity in pancreatic cancer cells, while SNHG14 knockdown exerted opposite effects. Mechanistic studies revealed that miR-613 was targeted by SNHG14, and Annexin A2 (ANXA2) was targeted and inversely regulated by miR-613 in pancreatic cancer cells. In vivo studies showed that SNHG14 knockdown attenuated tumour growth. MiR-613 was down-regulated and ANXA2 was up-regulated in the pancreatic cancer tissues, and SNHG14 expression levels were inversely correlated with miR-613 expression levels and positively correlated with the ANXA2 mRNA expression levels. Collectively, our results suggest that SNHG14 potentiates pancreatic cancer progression through modulation of annexin A2 expression via acting as a competing endogenous RNA for miR-613.

Laparoscopic Common Bile Duct Exploration Is a Safe and Effective Strategy for Elderly Patients.

PURPOSE: Laparoscopic common bile duct exploration (LCBDE) has been widely used to remove common bile duct (CBD) stones. However, surgery is not considered as the first treatment choice for elderly patients with CBD stones because of the potential risk of postoperative complications. This study aims to evaluate the safety and efficiency of LCBDE for elderly patients. METHODS: From April 2011 to October 2016, 265 consecutive patients underwent LCBDE. We performed a retrospective study and divided these patients into 2 groups. The younger group was younger than 70 years old (n = 179), and the elderly group was 70 years old or older (n = 86). We compared patient demographics, clinical characteristics, intraoperative parameters, postoperative complications, and incidence of recurrent stone between the 2 groups. RESULTS: The elderly patients had higher preoperative morbidity of chronic diseases, such as pulmonary diseases, heart diseases, arterial hypertension, and abdominal operation history ( P < .05). There were no significant differences between the 2 groups in terms of operation time, intraoperative blood loss, conversion rate to open surgery, total cost, overall complications, and incidence of recurrent stone ( P > .05). CONCLUSION: LCBDE can also be carried out as a safe and effective approach to remove CBD stones in elderly patients, although they have higher risk of chronic diseases.

Identification of Empoasca onukii (Hemiptera: Cicadellidae) and Monitoring of its Populations in the Tea Plantations of South China.

Tea green leafhoppers (Empoasca spp.) are considered one of the major pests in tea plantations in Asia. They are, however, difficult to monitor due to their size and flying and jumping abilities. In this study, we clarified the identification of the leafhopper species encountered in our study plantations and examined the impacts of sampling methods in estimating population abundance and sex ratio. The natural sex ratio of eggs, nymphs, and adults of tea green leafhopper and the differences between male and female were tested. Despite previous reports that Empoasca vitis (Goethe) was the major leafhopper present in our study area, our results showed that only Empoasca onukii Matsuda was found. Variation in population size over time and bias in sex ratio depending on the sampling methods were found in our monitoring experiments. In general, adult males were more attracted to yellow sticky cards than females. We believe that because female leafhoppers should be the target in pest control, yellow sticky cards may not be the most suitable monitoring or effective control of tea green leafhopper. We demonstrate the importance of understanding the implications of sampling techniques for population estimation and sex ratio bias as well as how temporal variation may affect monitoring results. Precise monitoring should take into consideration the different life histories of male and female.

Water-inrush mechanism from the head-on working face roof in a Jurassic coal seam in the Ordos Basin.

When Chinese coal mines are mining Carboniferous Permian coal seams, the mechanism of water inrush from the roof of the working face usually conforms to the "Upper Three Zones" or "Upper Four Zones" theory. The water inrush passageway is water-conducting fracture zone, and the water inrush position is located in the goaf. However, when mining Jurassic coal seams in Chinese coal mines, the location of water inrush often appears at the head-on working face, above the coal mining machine. Due to the support of the fully mechanized mining support, the roof rock layer cannot collapse and therefore cannot form water-conducting fracture zone. Therefore, the water inrush mechanism cannot be explained by the above two theories. This paper is guided by the Practical Mine Pressure Control Theory, and based on the explanation of the motion forms of bending (pulling) failure movement and shearing (cutting) failure movement, and combined with on-site examples, it is revealed that the passageways leading to from the head-on working face roof in a Jurassic coal seam in the Ordos Basin are splitting zones type, fracture line type and structural fracture type, respectively. Taking the changes in water inflow during the mining process of the 3301 and 3302 working faces in Zhujiamao Coal Mine as examples, this paper reveals the mechanism of water inrush from the head-on working face roof caused by splitting zones type, and proves the existence of this passageway through on-site 3D high-density electrical detection and tracing experiments. Taking two catastrophic water inrush accidents that occurred head-on in the 1309 working face of Guojiahe Coal Industry Co., Ltd. as examples, the water inrush mechanism of the fracture line type and the water inrush mechanism of the structural fracture type were respectively revealed. Based on mechanism of water inrush from head-on roof of working face and the analysis of the on-site water inrush process, a method for distinguishing the type of water inrush passageway from the front roof of the working face is proposed. The results indicate that the Jurassic coal seam mining in the Ordos Basin is prone to shearing (cutting) failure movement, resulting in the frequent formation of the three types of water inrush passageways mentioned above.

Application of Laennec extrathecal blockade combined with indocyanine green fluorescence imaging in laparoscopic anatomic hepatectomy.

OBJECTIVE: To investigate the safety and application value of combining Laennec extracapsular occlusion with ICG fluorescence imaging in laparoscopic anatomic hepatectomy. METHODS: Complete laparoscopic dissection was performed outside the Laennec sheath, blocking Glisson's pedicle of the corresponding liver segment or lobe. An appropriate amount of indocyanine green (ICG) dye was intravenously injected, and the boundary line between the pre-cut liver segment and liver lobe was identified using fluorescence laparoscopy. Complete resection of the liver segment or lobe was performed based on anatomical markers. Clinical data, including operation time, intraoperative blood loss, postoperative hospital stay, and postoperative complications, were collected. RESULTS: A total of 14 cases were included in the study, including seven cases of primary liver cancer, three cases of metastatic liver cancer, three cases of intrahepatic bile duct calculi, and one case of hepatic hemangioma. All 14 patients underwent anatomic hepatectomy under fluorescent laparoscopy, with four cases involving the right liver, seven cases involving the left liver, two cases involving the right anterior lobe, and one case involving the right posterior lobe. CONCLUSION: Combining laparoscopic follow-up of the Laennec membrane with Glisson outer sheath block and intraoperative ICG fluorescence imaging provides real-time guidance for locating the resection boundaries during anatomic hepatectomy. This approach helps in controlling intraoperative bleeding, reducing operation time, and ensuring high safety. It holds significant value in clinical application.

ULK1 Mediated Autophagy-Promoting Effects of Rutin-Loaded Chitosan Nanoparticles Contribute to the Activation of NF-κB Signaling Besides Inhibiting EMT in Hep3B Hepatoma Cells.

Background: Liver cancer remains to be one of the leading causes of cancer worldwide. The treatment options face several challenges and nanomaterials have proven to improve the bioavailability of several drug candidates and their applications in nanomedicine. Specifically, chitosan nanoparticles (CNPs) are extremely biodegradable, pose enhanced biocompatibility and are considered safe for use in medicine. Methods: CNPs were synthesized by ionic gelation, loaded with rutin (rCNPs) and characterized by ultraviolet-visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and transmission electron microscopy (TEM). The rCNPs were tested for their cytotoxic effects on human hepatoma Hep3B cells, and experiments were conducted to determine the mechanism of such effects. Further, the biocompatibility of the rCNPs was tested on L929 fibroblasts, and their hemocompatibility was determined. Results: Initially, UV-vis and FTIR analyses indicated the possible loading of rutin on rCNPs. Further, the rutin load was quantitatively measured using Ultra-Performance Liquid Chromatography (UPLC) and the concentration was 88 µg/mL for 0.22 micron filtered rCNPs. The drug loading capacity (LC%) of the rCNPs was observed to be 13.29 ± 0.68%, and encapsulation efficiency (EE%) was 19.55 ± 1.01%. The drug release was pH-responsive as 88.58% of the drug was released after 24 hrs at the lysosomal pH 5.5, whereas 91.44% of the drug was released at physiological pH 7.4 after 102 hrs. The cytotoxic effects were prominent in 0.22 micron filtered samples of 5 mg/mL rutin precursor. The particle size for the rCNPs at this concentration was 144.1 nm and the polydispersity index (PDI) was 0.244, which is deemed to be ideal for tumor targeting. A zeta potential (ζ-potential) value of 16.4 mV indicated rCNPs with good stability. The IC50 value for the cytotoxic effects of rCNPs on human hepatoma Hep3B cells was 9.7 ± 0.19 µg/mL of rutin load. In addition, the increased production of reactive oxygen species (ROS) and changes in mitochondrial membrane potential (MMP) were observed. Gene expression studies indicated that the mechanism for cytotoxic effects of rCNPs on Hep3B cells was due to the activation of Unc-51-like autophagy-activating kinase (ULK1) mediated autophagy and nuclear factor kappa B (NF-κB) signaling besides inhibiting the epithelial-mesenchymal Transition (EMT). In addition, the rCNPs were less toxic on NCTC clone 929 (L929) fibroblasts in comparison to the Hep3B cells and possessed excellent hemocompatibility (less than 2% of hemolysis). Conclusion: The synthesized rCNPs were pH-responsive and possessed the physicochemical properties suitable for tumor targeting. The particles were effectively cytotoxic on Hep3B cells in comparison to normal cells and possessed excellent hemocompatibility. The very low hemolytic profile of rCNPs indicates that the drug could be administered intravenously for cancer therapy.

Impact of Red Imported Fire Ant Nest-Building on Soil Properties and Bacterial Communities in Different Habitats.

The red imported fire ant (Solenopsis invicta Buren) is a highly adaptable invasive species that can nest and reproduce in different habitat soils. We aimed to explore the adaptability of red imported fire ants in different habitats by analyzing changes in the physicochemical properties of nest soils and bacterial communities. Five habitat types (forest, tea plantation, rice field, lawn, and brassica field) were selected. The results showed that the pH of the nest soils increased significantly in all five habitats compared to the control soils of the same habitat. A significant increase in nitrogen content was detected in the nests. The Cr, Pb, Cu, and Ni levels were significantly reduced in the soils of the five habitats, due to nesting activities. Analysis of the composition and diversity of the soil microbial community showed that, although the richness and diversity of bacteria in the nest soils of red imported fire ants in the five habitats varied, the relative abundance of Actinobacteria significantly increased and it emerged as the dominant bacterial group. These results indicate that red imported fire ants modify the physicochemical properties of nest soils and bacterial communities to create a suitable habitat for survival and reproduction.

Preventing water-inrush from floor in coal working face with paste-like backfill technology.

In the view of the situation where great economic loss often occurs during mining deep coal seams in Feicheng coal field due to water inrush from the floor Ordovician limestone aquifer, the floor "lower four-zone" theory was used as a guide. 81006 working face of Caozhuang Coalmine in Feicheng coal field was taken as the research background, and paste filling technology was proposed to inhibit or reduce the damaged floor depth caused by mine ground pressure in order to prevent water inrush from the floor Ordovician limestone aquifer. Glue material, coal gangue powder, and fly ash were selected as filling material, and a ground filling system, including a material production system, storage material system, power supply and water supply system, automatic control and measurement system, monitoring and control communications system, emergency response system, and underground filling system, including pipe conveyor system and working face blocking grout loss system, were established to achieve effective filling goaf. Field stress monitoring and floor damaged depth measurement showed that when reaching a steady state after a period of time, paste filling working face not only restored to the original stress state but also significantly reduced the floor's damaged depth caused by mine ground pressure. This verifies that working face with paste-like backfill technology is a very effective measure to prevent water-inrush from the floor.

The influences of green light on locomotion, growth and reproduction in the brown planthopper Nilaparvata lugens.

BACKGROUND: Light stimulation at a specific wavelength triggers various responses in insects and can be used for pest control. To develop efficient and ecofriendly photophysical pest control methods, the effects of green light on locomotion, growth (molting and eclosion) and reproduction in Nilaparvata lugens (Stål) (BPH), a major rice pest, were studied. Transcriptomics and transmission electron microscopy (TEM) were used to investigate the mechanisms involved. RESULTS: BPH adults showed disrupted daily locomotion patterns following green light treatment at night and exhibited abnormal locomotion peaks. Total 6-day locomotion of brachypterous adults was significantly greater than in the control group. The durations of growth stages 1-4 were all shorter under green light treatment than in the control, whereas the time from fourth molting to eclosion (stage 5) was significantly longer. When BPH adults under green light treatment began laying eggs, the egg hatching ratio (36.69%) was significantly lower than in the control (47.49%). Moreover, in contrast to the control, BPH molting and eclosion events tended to happen more at night. Transcriptome analysis proved that green light significantly affected the expression of genes involved in cuticular proteins, chitin deacetylase and chitinase, which are related to cuticular development. TEM observations confirmed abnormal cuticular development in nymph and adult BPHs (endocuticle, exocuticle and pore canals) under green light treatment. CONCLUSION: Green light treatment at night notably affected locomotion, growth and reproduction in BPH, thus providing a novel idea for controlling this pest. © 2023 Society of Chemical Industry.

Shifts in the diversity of root endophytic microorganisms across the life cycle of the ratooning rice Jiafuzhan.

The diversity of root endophytic microorganisms, which is closely related to plant life activities, is known to vary with the plant growth stage. This study on the ratooning rice Jiafuzhan explored the diversity of the root endophytic bacteria and fungi and their dynamics during the plant life cycle. By sequencing the 16S ribosomal ribonucleic acid (16S rRNA) and internal transcribed spacer (ITS) genes, 12,154 operational taxonomic units (OTUs) and 497 amplicon sequence variants (ASVs) were obtained, respectively. The root endophytic microorganisms of rice in the seedling, tillering, jointing, heading, and mature stages of the first crop and at 13, 25, and 60 days after regeneration (at the heading, full heading, and mature stages of the second crop, respectively) were analyzed using diversity and correlation analyses. There were significant differences in the α-diversity and ß-diversity of root endophytic bacteria and fungi in the growth stage. Additionally, linear discriminant analysis (LDA) effect size (LEfSe) analysis revealed biomarker bacteria for each growth stage, but biomarker fungi did not exist in every stage. Moreover, the correlation analysis showed that the bacterial and fungal biomarkers interacted with each other. Furthermore, the nitrogen-fixing genus Bradyrhizobium existed in all growth stages. These findings indicate the pattern of root endophytic microorganisms of ratooning rice at different growth stages, and they provide new insights into the high yield of the second crop of ratooning rice (in light of the abundance of various bacteria and fungi).

CD8+iTregs mediate the protective effect of rapamycin against graft versus host disease in a humanized murine model.

CD8+Tregs are important immunoregulatory cells that participate in immunopathological processes in many diseases. Rapamycin (Rapa) is a macrolide immunosuppressant that inhibits the mammalian target of rapamycin (mTOR) and has been shown to improve CD4+-induced Tregs (iTregs) generation. This study aimed to evaluate the role of Rapa in the generation and function of CD8+iTregs. Human CD8 + CD25-CD45RA + T cells were divided into two groups, one with Rapa and the other without Rapa, and both groups were cultured under Treg-induced conditions. Rapa significantly improved Foxp3 expression and the suppressive function of CD8+iTregs in vitro. Further studies showed that Rapa suppressed inflammatory cytokine expression and enhanced anti-inflammatory cytokine expression. Under inflammatory conditions in vitro, Rapa-CD8 + iTregs sustained Foxp3 and anti-inflammatory cytokine expression. An in-depth study showed that Rapa regulated CpG demethylation in the Foxp3 region and STAT1 and STAT3 phosphorylation in CD8+iTregs. Finally, we compared the regulatory ability of Rapa and all-trans retinoic acid, another reagent that stimulates CD4+ iTreg generation in vitro, which showed that Rapa, but not all-trans retinoic acid, improved CD8+ iTreg induction and suppressed CD4+T cell expansion in vitro and protected against graft-versus-host disease in a humanized murine model in vivo. These results strongly suggest that CD8+iTregs initiated by Rapa may represent a new therapeutic strategy for inflammatory and autoimmune diseases.

Leaching characteristics and pollution risk assessment of potentially harmful elements from coal gangue exposed to weathering for different periods of time.

To explore the leaching behavior and potential degree of pollution that can result from the backfilling of goafs with different types of coal gangue (CG), fresh CG from the Hongqi Coal Mine goaf and surface CG (weathered for 1 year) were selected as the research objects in this study. A series of leaching experiments were carried out using the Ordovician limestone karst waters of the mining areas as the soaking solution. A comparative study on the dissolution characteristics of Fe3+, Mn2+, and SO42- and on the traditional water quality parameters of the two types of CG was conducted. The results showed that the soaked, weathered CG displayed a higher ion dissolution value than fresh CG. The ratio of each ion was as follows: Fe3+ was 1, Mn2+ was 2.86 ~ 68.18, and SO42- was 1.34 ~ 2.09. Over time, the ion concentration of water samples that initially contained high ion concentration values showed a decreasing trend after CG was soaked in these waters, but the values were still in the range of high ion release concentrations. The pH and oxidation‒reduction potential (ORP) values of the leachate of both CG types indicated that the leachates were weakly alkaline and weakly oxidizing, and the overall change in total dissolved solids (TDS) was small and consistent with the SO42- trend. SO42- in the leachate of the weathered CG showed a more significant correlation with the pH and TDS of the soaking solution, and it was the major pollutant. According to the geoaccumulation index evaluation, weathered CG had higher pollution potential than fresh CG. Fe3+ presented a slight and moderate risk for contamination.

Analysis of Related Risk Factors of Microvascular Invasion in Hepatocellular Carcinoma.

Objective: To forecast the onset of microvascular invasion (MVI) in patients with hepatoma by evaluating the preoperative aspartate aminotransferase-to-platelet ratio index (APRI), alpha-fetoprotein (AFP), neutrophil-to-lymphocyte ratio (NLR), and other clinicopathological data. Methods: In this study, we retrospectively analysed the clinical data of 62 patients who received radical surgery for hepa toma from 2019 to 2021. Patients were separated into the MVI-negative group and the MVI-positive group according to the postoperative pathological diagnosis. The relationships between MVI and NLR, APRI, AFP, tumor size, and other clinical data were assessed using the univariate analysis, receiver operating characteristic (ROC) curve, least absolute shrinkage and selection operator (LASSO) analysis, and logistic analysis. Results: The ROC curve determined that the cutoff values of NLR, platelet-to-lymphocyte ratio (PLR), and APRI were 1.520, 98, and 0.275, respectively. The univariate analysis showed that the MVI-positive result was associated with five factors: tumor size (χ 2 = 10.620, p = 0.001), AFP (χ 2 = 10.524, p = 0.001), Edmondson grade (χ 2 = 20.736, p < 0.001), NLR (χ 2 = 8.744, p = 0.003), and APRI (χ 2 = 4.849, p = 0.028). The LASSO analysis indicated that the risk factors were the number of tumors, PLR, APRI, NLR, AFP, Edmondson grade, and tumor size. The multivariate logistic regression analysis showed that NLR ≥ 1.520 (OR 11.119, p = 0.006), APRI ≥ 0.275 (OR 12.515, p = 0.009), AFP ≥ 200 µg/mL (OR 7.823, p = 0.016), and tumor size > 3 cm (OR 7.689, p = 0.022) were independent risk factors for MVI in patients with hepatoma. Conclusion: Preoperative NLR, APRI, AFP, and tumor size are reliable indicators for predicting the appearance of MVI in patients with hepatoma and are of great value in making detailed and reliable treatment protocols for these patients before surgery.

Evaluation of the Parasitism Capacity of a Thelytoky Egg Parasitoid on a Serious Rice Pest, Nilaparvata lugens (Stål).

Pseudoligosita yasumatsui and Anagrus nilaparvatae are both egg parasitoids of the brown planthopper, Nilaparvata lugens (Stål) (BPH). In this study, we obtained a stable strain of P. yasumatsui reproduced via thelytoky through indoor rearing and screening. We assessed the parasitism capacity of this strain on eggs of N. lugens by comparing the parasitism preference and circadian rhythm of this strain to that of A. nilaparvatae, which is proved as the dominant egg parasitoid species of BPH in rice fields. The findings indicated that both egg parasitoids could parasitize fertilized and unfertilized BPH eggs, however, with a significant preference for fertilized eggs. The daily parasitization volume of P. yasumatsui was slightly higher than that of A. nilaparvatae. Both egg parasitoids preferred parasitizing 1-3-day-old BPH eggs, but the parasitism amount of 5-6-day-old BPH eggs by P. yasumatsui is higher than that by A. nilaparvatae. The parasitism events of both species of egg parasitoid wasps occurred primarily from 7:00-15:00 and the parasitism amount at night accounted for less than 15% of the total amount. The results indicate that this strain of P. yasumatsui reproduced via thelytoky could be valuable for rice planthopper control.