Simultaneous profiling of host expression and microbial abundance by spatial metatranscriptome sequencing.

We developed an analysis pipeline that can extract microbial sequences from spatial transcriptomic (ST) data and assign taxonomic labels, generating a spatial microbial abundance matrix in addition to the default host expression matrix, enabling simultaneous analysis of host expression and microbial distribution. We called the pipeline spatial metatranscriptome (SMT) and applied it on both human and murine intestinal sections and validated the spatial microbial abundance information with alternative assays. Biological insights were gained from these novel data that showed host-microbe interaction at various spatial scales. Finally, we tested experimental modification that can increase microbial capture while preserving host spatial expression quality and, by use of positive controls, quantitatively showed the capture efficiency and recall of our methods. This proof-of-concept work shows the feasibility of SMT analysis and paves the way for further experimental optimization and application.

Blue Light Regulates Cell Wall Structure and Carbohydrate Metabolism of Soybean Hypocotyl.

Soybean stem elongation and thickening are related to cell wall composition. Plant morphogenesis can be influenced by blue light, which can regulate cell wall structure and composition, and affect stem growth and development. Here, using proteomics and metabolomics, differentially expressed proteins and metabolites of hypocotyls grown in the dark and under blue light were studied to clarify the effects of blue light on the cell wall structure and carbohydrate metabolism pathway of soybean hypocotyls. Results showed that 1120 differential proteins were upregulated and 797 differential proteins were downregulated under blue light treatment, while 63 differential metabolites were upregulated and 36 differential metabolites were downregulated. Blue light promoted the establishment of cell wall structure and composition by regulating the expression of both the enzymes and metabolites related to cell wall structural composition and nonstructural carbohydrates. Thus, under blue light, the cross-sectional area of the hypocotyl and xylem were larger, the longitudinal length of pith cells was smaller, elongation of the soybean hypocotyl was inhibited, and diameter was increased.

Safety assessment and probiotic characteristics of Enterococcus lactis JDM1.

OBJECTIVE: The aims of this study were to evaluate the safety and probiotic characteristics of the newly isolated Enterococcus lactis strain JDM1. METHODS: Safety assessment of E. lactis JDM1 was accomplished by the combination of whole genome sequence information analysis and phenotypic assays, including antimicrobial susceptibility test, haemolysis assay, biogenic amine production assay, cytotoxicity assay. The bacteriostatic experiment and gastrointestinal tolerance experiment were also conducted to evaluate its applicability. RESULTS: E. lactis JDM1 possesses good gastrointestinal tolerance and can inhibit the growth of the pathogenic bacteria Clostridioides difficile and Listeria monocytogenes. The chromosome size of JDM1 was 2,570,998 bp with a GC content of 38.46%, which contained a plasmid. One intact prophage, 13 genomic islands and 19 IS elements were predicted in the JDM1 chromosome. Five resistance-related genes and seven virulence-related genes were predicted in the genome. Most resistance genes were conserved, and virulence factors were not related to functional pathogenicity. Antimicrobial susceptibility tests showed that JDM1 was sensitive to tedizolid, ciprofloxacin, levofloxacin, penicillin, ampicillin, vancomycin, linezolid, tetracycline, high-level gentamicin and high-level streptomycin. Genes encoding putative enzymes responsible for adverse metabolites were not found and JDM1 was unable to produce the six main biogenic amines. Cytotoxicity test showed that the JDM1 supernatant had no toxic effect. CONCLUSION: E. lactis JDM1 is expected to be developed as a probiotic, and its probiotic properties are worthy of further exploration.

A newly isolated E. thailandicus strain d5B with exclusively antimicrobial activity against C. difficile might be a novel therapy for controlling CDI.

Colitis induced by C. difficile is one of the most common and costly healthcare-related infections for humans. Probiotics are one of the most promising approaches for controlling CDI. Here, we presented the isolation, safety, and probiotic property evaluation of a novel E. thailandicus strain, d5B, with effective antimicrobial activity against C. difficile. Strain d5B showed strong bactericidal effects on at least 54C. difficile strains. Safety tests showed that strain d5B was sensitive to clinically important antibiotics, and had no haemolytic and cytotoxic activities. Whole genomic analysis showed strain d5B only contained one aminoglycoside resistance gene located in the chromosome. Moreover, d5B was devoid of functional virulence genes. Finally, strain d5B exhibited probiotic properties, such as tolerance to the gastrointestinal tract, and adhered well to HT-29 cells. In conclusion, the E. thailandicus strain d5B should be investigated further for useful properties as a novel candidate probiotic for controlling CDI.

[Research progress of vascular endothelial growth factor-A and its isoforms in kidney disease].

Vascular endothelial growth factor-A (VEGF-A) is a critical angiogenic factor which is mainly secreted from podocytes and epithelial cells in kidney and plays an important role in renal pathophysiology. In recent years, functions of different isoforms of VEGF-A and the new secretion approach via extracellular vesicles (EVs) have been identified. Thus, further understanding are needed for the role of VEGF-A and its isoforms in renal injury and repair. In this review, we summarized the expression, secretion and regulation of VEGF-A, its biological function, and the role of different isoforms of VEGF-A in the development of different renal diseases. Meanwhile, the research progress of VEGF-A as diagnostic marker and therapeutic target for renal diseases were discussed.

[Research progress of extracellular vesicles in the treatment of renal disease].

Extracellular vesicles (EVs) are lipid bilayer-enclosed structures containing diverse bioactive cargoes that play a major role in intercellular communication in both physiological and pathological conditions. Currently, the field of EV-based therapy has been rapidly growing, and two main therapeutic uses of EVs can be surmised: (i) exploiting stem cell-derived EVs as therapeutic agents; and (ii) employing EVs as natural therapeutic vectors for drug delivery. This review will discuss the recent advances in EV-based therapy in the treatment of renal disease.

Modifiable risk factors of acute kidney injury after liver transplantation: a systematic review and meta-analysis.

BACKGROUND: Acute kidney injury (AKI) is a common and critical complication of liver transplantation (LT), which is associated with increased morbidity, mortality and health care cost. We aimed to identify modifiable risk factors of AKI after LT. METHODS: A literature search of Pubmed, EMBASE and Cochrane Databases was performed to identify studies investigating risk factors of AKI after LT. The Newcastle-Ottawa Scale was used to rate study quality. Effect size and 95% confidence interval were pooled using a random-effect model with inverse-variance method. RESULTS: Sixty-seven articles with 28,844 patients were included in the meta-analysis. Seventeen modifiable risk factors were found, including overweight, preoperative use of diuretic, preoperative anemia, donation after cardiac death organ, donor BMI ≥ 30 kg/m2, ABO-incompatible LT, low graft to recipient body weight ratio, intraoperative hypotension, major bleeding, intraoperative use of vasopressor, large RBC transfusion, postreperfusion syndrome, postoperative use of vasopressors, overexposure to calcineurin inhibitor, calcineurin inhibitor without mycophenolate mofetil, graft dysfunction and infection. A total of 38 articles were included in the systematic review, in which 8 modifiable risk factors and 1 protective factor were additionally associated in single studies with the incidence of AKI after LT. CONCLUSIONS: Effective interventions based on identified modifiable risk factors in the perioperative management and graft allocation and preservation may be promising to reduce the incidence of AKI after LT. TRIAL REGISTRATION: The protocol for this systematic review is registered with PROSPERO (No. CRD42020166918 ).

Effects of inner muscle resistance exercise on stress urinary incontinence: a randomized clinical controlled trial.

[Purpose] This study investigated the effects of co-contraction resistance exercises of the transverse abdominal and pelvic floor muscles in middle-aged females with stress urinary incontinence. [Participants and Methods] We included 32 females with stress urinary incontinence and divided them into two groups: the inner muscle training group and the pelvic floor muscle group. The thickness of the transverse abdominal muscle was measured during four tasks: (1) rest, (2) maximum contraction of the transverse abdominal muscle, (3) maximum contraction of the pelvic floor muscle, and (4) maximum co-contraction of the transverse abdominal and pelvic floor muscles. In the latter three tasks, measurements were obtained while the participants performed resistance movements using a Thera-band®. A home program was conducted in both groups, and the intervention lasted for 8 weeks. [Results] The cure rates for SUI were 87.5% and 68.8% in the inner muscle training and pelvic floor muscle groups, respectively. After the intervention, the thickness of the transverse abdominal muscle significantly increased in the inner muscle training groups performing maximum co-contraction of the transverse abdominal and pelvic floor muscles and maximum contraction of the transverse abdominal muscle. [Conclusion] Inner muscle training exercises are more effective than pelvic floor muscle exercises in improving inner muscle function and urinary incontinence in middle-aged females.

Measurement reliability and cooperative movement of the pelvic floor and transverse abdominal muscles.

[Purpose] This study examined the measurement reliability and cooperative movement of the pelvic floor and transverse abdominal muscles. [Participants and Methods] The participants were seven healthy adult females. Transverse abdominal muscle thickness and bladder floor elevation were measured under the following conditions during active exercise and during resistance exercise: the resting state, maximum contraction of the transverse abdominal muscle, maximum contraction of the pelvic floor muscle, and maximum co-contraction of the transverse abdominal and pelvic floor muscles. Measurements were taken at rest and under each exercise condition. [Results] The intraclass correlation coefficients of transverse abdominal muscle thickness and bladder floor elevation showed high reproducibility under all conditions. The maximum contraction of the pelvic floor muscle showed a high correlation with the maximum co-contraction of the transverse abdominal muscle and pelvic floor muscle during resistance exercise. A significant regression line was found between transverse abdominal muscle thickness and bladder floor elevation under all conditions. The regression equation was as follows: transverse abdominal muscle thickness=0.113 bladder floor elevation+0.377 (r2=0.21). [Conclusion] This study demonstrated that the measurement reliability of the transverse abdominal and pelvic floor muscles is high, and that both muscles exhibit cooperative movement.

Association of overweight with postoperative acute kidney injury among patients receiving orthotopic liver transplantation: an observational cohort study.

BACKGROUND: Acute kidney injury (AKI) is a common postoperative complication of orthotopic liver transplantation (OLT). So far, little attention has been paid on the association between overweight and AKI after OLT, and animal models or clinical studies have drawn conflicting conclusions. The objective of our study was to determine whether overweight (BMI [Body Mass Index] ≥ 25 kg/m2) is associated with an increased risk of AKI after OLT. METHODS: This retrospective cohort study included 244 patients receiving OLT in the Affiliated Hospital of Qingdao University between January 1, 2017, and August 29, 2019. Preoperative, intraoperative, and postoperative data were collected retrospectively. The primary outcome was the development of AKI as defined by Kidney Disease, Improving Global Outcome (KIDGO) staging system. Logistic regression analysis was used to determine the relationship between overweight and the occurrence of postoperative AKI. Data analysis was conducted from September to October 2019, revision in April 2020. RESULTS: Among 244 patients receiving OLT (mean [standard deviation] age, 54.1 [9.6] years; 84.0% male) identified, 163 patients (66.8%) developed postoperative AKI. Overweight (BMI ≥ 25 kg/m2) was associated with a higher rate of postoperative severe AKI (stage 2/3) compared with normal weight (18.5 ≤ BMI < 25 kg/m2) (41 [47.7%] vs 39 [28.7%]; adjusted odds ratio [OR], 2.539; 95% confidence interval [CI], 1.389-4.642; P = 0.002). Furthermore, patients with obese were at even higher risk of postoperative severe AKI after controlling for confounding factors (adjusted OR: 3.705; 95% CI: 1.108-12.388; P = 0.033). CONCLUSIONS: Overweight is independently associated with an increased risk of postoperative severe AKI among patients receiving OLT. The association of BMI with severe AKI after OLT is J-shaped.

Association of intraoperative hypotension with acute kidney injury after liver resection surgery: an observational cohort study.

BACKGROUND: Acute kidney injury (AKI) is a major complication following liver resection. The aim of this study was to determine the risk factors for AKI after hepatic resection and whether intraoperative hypotension (IOH) was related to AKI. METHODS: Adult patients (≥ 18 years) undergoing liver resection between November 2017 and November 2019 at our hospital were retrospectively reviewed. AKI was defined as ≥50% increase in serum creatinine from baseline value within 48 h after surgery. IOH was defined as the lowest absolute mean arterial pressure (MAP) < 65 mmHg for more than 10 cumulative minutes during the surgery. Patients were divided into AKI group and non-AKI group, and were stratified by age ≥ 65 years. RESULTS: 796 patients who met our inclusion and exclusion criteria were analyzed. After multivariable regression analysis, the IOH (OR, 2.565; P = 0.009) and age ≥ 65 years (OR, 2.463; P = 0.008) were risk factors for AKI. The IOH (OR, 3.547; P = 0.012) and received red blood cell (OR, 3.032; P = 0.036) were risk factors of AKI in age ≥ 65 years patients. CONCLUSIONS: The IOH and age ≥ 65 years were risk factors for postoperative AKI, and IOH was associated with AKI in age ≥ 65 years patients following liver resection.

Selective export of autotaxin from the endoplasmic reticulum.

Autotaxin (ATX) or ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2) is a secretory glycoprotein and functions as the key enzyme for lysophosphatidic acid generation. The mechanism of ATX protein trafficking is largely unknown. Here, we demonstrated that p23, a member of the p24 protein family, was the protein-sorting receptor required for endoplasmic reticulum (ER) export of ATX. A di-phenylalanine (Phe-838/Phe-839) motif in the human ATX C-terminal region was identified as a transport signal essential for the ATX-p23 interaction. Knockdown of individual Sec24 isoforms by siRNA revealed that ER export of ATX was impaired only if Sec24C was down-regulated. These results suggest that ATX is selectively exported from the ER through a p23, Sec24C-dependent pathway. In addition, it was found that AKT signaling played a role in ATX secretion regulation to facilitate ATX ER export by enhancing the nuclear factor of activated T cell-mediated p23 expression. Furthermore, the di-hydrophobic amino acid motifs (FY) also existed in the C-terminal regions of human ENPP1 and ENPP3. Such a p23, Sec24C-dependent selective ER export mechanism is conserved among these ENPP family members.

Incidence, Risk Factors, and Outcomes of Hyperbilirubinemia in Adult Cardiac Patients Supported by Veno-Arterial ECMO.

The aims of this study were to evaluate the incidence, risk factors, and outcomes of hyperbilirubinemia in cardiac patients with veno-arterial (VA) ECMO. Data on 89 adult patients with cardiac diseases who received VA ECMO implantation in our hospital were retrospectively reviewed. All patients were divided into the following three groups: 24 in normal group (N, total bilirubin [TBIL] ≤3 mg/dL), 30 in high bilirubin group (HB, 6 mg/dL ≥ TBIL > 3 mg/dL), and 35 in severe high bilirubin group (SHB, TBIL > 6 mg/dL). lg(variables + 1) was performed for nonnormally distributed variables. The incidence of hyperbilirubinemia (>3 mg/dL) was 73%. In a multiple linear regression analysis, lg(peak TBIL + 1) was significantly associated with lg(peak AST + 1) (b-coefficient 0.188, P = 0.001), lg(peak pFHb + 1) (b-coefficient 0.201, P = 0.003), and basic TBIL (b-coefficient 0.006, P = 0.009). Repeated measurement analysis of variance revealed that the main effect for three groups in pFHb and lg(AST + 1) was significant at first 3 days during ECMO. The patients in SHB had low platelets during ECMO and low in-hospital survival rate. Hyperbilirubinemia remains common in patients with VA ECMO and is associated with low platelets and high in-hospital mortality. Hemolysis and liver dysfunction during ECMO and basic high bilirubin levels are risk factors of hyperbilirubinemia.

[Urinary biomarkers for chronic kidney disease: a focus on gene transcript].

Chronic kidney disease (CKD) is becoming an alarming health burden worldwide, however, there is still lack of early biomarkers and effective treatment options. Thus, in the upcoming era of precision medicine, searching for the sensitive, non-invasive biomarkers has been the cornerstone and major challenge in the management of CKD. Urine contains rich biological information which could be an ideal source for non-invasive biomarkers of CKD. This review will discuss the recent advances in biomarker study from urine sediment, urine supernatant and urinary extracellular vesicles with special interest in gene transcript (miRNA, mRNA) biomarkers. Besides, the challenges and future directions for urinary gene transcript biomarker study will be discussed.

Autotaxin Expression Is Regulated at the Post-transcriptional Level by the RNA-binding Proteins HuR and AUF1.

Autotaxin (ATX) is a key enzyme that converts lysophosphatidylcholine (LPC) into lysophosphatidic acid (LPA), a lysophospholipid mediator that regulates cellular activities through its specific G protein-coupled receptors. The ATX-LPA axis plays an important role in various physiological and pathological processes, especially in inflammation and cancer development. Although the transcriptional regulation of ATX has been widely studied, the post-transcriptional regulation of ATX is largely unknown. In this study, we identified conserved adenylate-uridylate (AU)-rich elements in the ATX mRNA 3'-untranslated region (3'UTR). The RNA-binding proteins HuR and AUF1 directly bound to the ATX mRNA 3'UTR and had antagonistic functions in ATX expression. HuR enhanced ATX expression by increasing ATX mRNA stability, whereas AUF1 suppressed ATX expression by promoting ATX mRNA decay. HuR and AUF1 were involved in ATX regulation in Colo320 human colon cancer cells and the LPS-stimulated human monocytic THP-1 cells. HuR knockdown suppressed ATX expression in B16 mouse melanoma cells, leading to inhibition of cell migration. This effect was reversed by AUF1 knockdown to recover ATX expression or by the addition of LPA. These results suggest that the post-transcriptional regulation of ATX expression by HuR and AUF1 modulates cancer cell migration. In summary, we identified HuR and AUF1 as novel post-transcriptional regulators of ATX expression, thereby elucidating a novel mechanism regulating the ATX-LPA axis.

Plasma Free Hemoglobin Is a Predictor of Acute Renal Failure During Adult Venous-Arterial Extracorporeal Membrane Oxygenation Support.

OBJECTIVE: Hemolysis is a common and severe complication during extracorporeal membrane oxygenation (ECMO). Increased plasma free hemoglobin (PFHb) is related to renal injury. The aim of this study was to investigate whether increased PFHb during adult venous-arterial ECMO was associated with acute renal failure (ARF). DESIGN: A retrospective, observational, single-center study. SETTING: Fuwai Hospital in Beijing, China. PARTICIPANTS: The study comprised 84 venous-arterial ECMO patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 84 consecutive adult patients (≥18 years) with cardiac diseases requiring venous-arterial ECMO support were studied retrospectively. Demographics of patients, clinical and ECMO characteristics, and PFHb level were collected within the first 3 days after ECMO. ARF was defined as a≥300% rise in serum creatinine from baseline or application of dialysis. Repeated measurement analysis of variance revealed that the main effect for the non-ARF group and ARF group in PFHb (p = 0.002) was significant. A significant main effect for time points (p<0.001) and time×group interaction (p = 0.014) in PFHb was obtained. In a multiple logistic regression model, peak PFHb during ECMO (odds ratio 1.052, 95% confidence interval 1.016-1.089, p = 0.005) was a risk factor for ARF during ECMO and patients who underwent heart transplantation (odds ratio 0.240, 95% confidence interval 0.060-0.964, p = 0.044) experienced less ARF. There was a linear correlation between peak serum creatinine and peak PFHb (Spearman's r = 0.223, p = 0.042). CONCLUSIONS: Increased PFHb is a predictor of ARF among adult patients on venous-arterial ECMO support.

[Effect of aquaporin 9 on proliferation, apoptosis, invasiveness and migration of HepG2 cells].

OBJECTIVE: To investigate the impact of aquaporin 9 (AQP9) on the proliferation,apoptosis,invasiveness and migration of hepatocellular carcinoma cells using the HepG2 cell line. METHODS: A lentiviral vector targeting the coding region of human AQP9 was constructed. The recombinant lentiviral vector was harvested from the 293T cell line and transfected into the HepG2 cell line; resistant cell clones were selected with puromycin. Three groups of cells were established, including the CC group (control without lentiviral vector), the PWPI group (control with empty carrier virus), and the AQP9 overexpression group (experimental with the AQP9 recombinant virus). Transfection efficiency was validated by laser confocal microscopy.Expression of AQP9 was detected in the transfected HepG2 cells by westem blotting (protein) and real-time qPCR (mRNA). AQP9 effects on proliferation, migration, invasion and apoptosis of the HepG2 cell line were assessed by plate colony formation assay, woumd healing assay, transwell assay and flow cytometry. RESULTS: The green fluorescent protein of the recombinant lentiviral vector was appropriately distributed in the cell membrane. The AQP9 overexpression group showed significantly higher AQP9 mRNA and protein levels than the PWPI group and the CC group (both P < 0.01). Cells with AQP9 overexpression showed a lower colony formation rate (16.93±3.19% vs. CC group: 23.53±2.10% and PWPI group: 23.00±2.02%; F=6.46, P=0.032) and a lower overall apoptosis rate (44.96±3.53% vs. CC group:19.7±2.49% and PWPI group: 24.37±2.38%; F=66.88, P < 0.01). The AQP9 overexpression group also showed significantly higher number of cells in the G1 stage and significantly lower number of cells in the S stage (G1: 66.58±0.99% and S:15.25±1.81%), significantly smaller cell migration distance (P=0.01 < 0.05), and significantly suppressed invasiveness (17±8 vs. CC group:109+/-9 and PWPI group: 95±11; P=0.01 < 0.05). CONCLUSION: In HepG2 cells, AQP9 significantly reduces the migrative and invasive capabilities, induces cell apoptosis, and inhibits cell proliferation via cell cycle arrest at the G1/S phases.

Identification of the biological functions and chemo-therapeutic responses of ITGB superfamily in ovarian cancer.

BACKGROUND: Patients with ovarian cancer (OC) tend to face a poor prognosis due to a lack of typical symptoms and a high rate of recurrence and chemo-resistance. Therefore, identifying representative and reliable biomarkers for early diagnosis and prediction of chemo-therapeutic responses is vital for improving the prognosis of OC. METHODS: Expression levels, IHC staining, and subcellular distribution of eight ITGBs were analyzed using The Cancer Genome Atlas (TCGA)-Ovarian Serous Cystadenocarcinoma (OV) database, GEO DataSets, and the HPA website. PrognoScan and Univariate Cox were used for prognostic analysis. TIDE database, TIMER database, and GSCA database were used to analyze the correlation between immune functions and ITGBs. Consensus clustering analysis was performed to subtype OC patients in the TCGA database. LASSO regression was used to construct the predictive model. The Cytoscape software was used for identifying hub genes. The 'pRRophetic' R package was applied to predict chemo-therapeutic responses of ITGBs. RESULTS: ITGBs were upregulated in OC tissues except ITGB1 and ITGB3. High expression of ITGBs correlated with an unfavorable prognosis of OC except ITGB2. In OC, there was a strong correlation between immune responses and ITGB2, 6, and 7. In addition, the expression matrix of eight ITGBs divided the TCGA-OV database into two subgroups. Subgroup A showed upregulation of eight ITGBs. The predictive model distinguishes OC patients from favorable prognosis to poor prognosis. Chemo-therapeutic responses showed that ITGBs were able to predict responses of common chemo-therapeutic drugs for patients with OC. CONCLUSIONS: This article provides evidence for predicting prognosis, immuno-, and chemo-therapeutic responses of ITGBs in OC and reveals related biological functions of ITGBs in OC.

WITHDRAWN: TRIM44 promotes autophagy through SQSTM1 oligomerization in the response to oxidative stress induced by Arsenic Trioxide in cancer cells.

The authors have requested that this preprint be removed from Research Square.

Nanostructured SnO2 Thin Films Based on a Convenient Chemical Deposition for Sensitive Detection of Ethanol.

A specific matrix sensor that can operate at low temperatures and has a high sensing response is crucial for monitoring flammable VOC gases. In this study, a nanostructured SnO2 thin film was successfully produced using a suitable chemical deposition method, and its sensing properties were comprehensively analyzed. The SEM images revealed that the thin film of the nanostructured SnO2 is made up of two different sizes of broccoli-like structure nanoparticles. The sensor, which is based on this unique micronano structure, demonstrated a high sensing response (44), low operating temperature (200 °C), and fast response time (6s). Additionally, the nanostructured sensor exhibited excellent resistance to humidity interference and long-term stability. Moreover, DFT is employed to evaluate the electronic properties and to systematically explain the gas sensing mechanism of the nanostructured sensor based on the SnO2 thin film.