CSMD3 Deficiency Leads to Motor Impairments and Autism-Like Behaviors via Dysfunction of Cerebellar Purkinje Cells in Mice.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder with highly heritable heterogeneity. Mutations of CUB and sushi multiple domains 3 (CSMD3) gene have been reported in individuals with ASD. However, the underlying mechanisms of CSMD3 for the onset of ASD remain unexplored. Here, using male CSMD3 knock-out (CSMD3 -/-) mice, we found that genetic deletion of CSMD3 produced core autistic-like symptoms (social interaction deficits, restricted interests, and repetitive and stereotyped behaviors) and motor dysfunction in mice, indicating that the CSMD3 gene can be considered as a candidate for ASD. Moreover, we discovered that the ablation of CSMD3 in mice led to abnormal cerebellar Purkinje cell (PC) morphology in Crus I/II lobules, including aberrant developmental dendritogenesis and spinogenesis of PCs. Furthermore, combining in vivo fiber photometry calcium imaging and ex vivo electrophysiological recordings, we showed that the CSMD3 -/- mice exhibited an increased neuronal activity (calcium fluorescence signals) in PCs of Crus I/II lobules in response to movement activity, as well as an enhanced intrinsic excitability of PCs and an increase of excitatory rather than inhibitory synaptic input to the PCs, and an impaired long-term depression at the parallel fiber-PC synapse. These results suggest that CSMD3 plays an important role in the development of cerebellar PCs. Loss of CSMD3 causes abnormal PC morphology and dysfunction in the cerebellum, which may underlie the pathogenesis of motor deficits and core autistic-like symptoms in CSMD3 -/- mice. Our findings provide novel insight into the pathophysiological mechanisms by which CSMD3 mutations cause impairments in cerebellar function that may contribute to ASD.SIGNIFICANCE STATEMENT Autism spectrum disorder (ASD) is a neurodevelopmental disorder with highly heritable heterogeneity. Advances in genomic analysis have contributed to numerous candidate genes for the risk of ASD. Recently, a novel giant gene CSMD3 encoding a protein with CUB and sushi multiple domains (CSMDs) has been identified as a candidate gene for ASD. However, the underlying mechanisms of CSMD3 for the onset of ASD remain largely unknown. Here, we unravel that loss of CSMD3 results in abnormal morphology, increased intrinsic excitabilities, and impaired synaptic plasticity in cerebellar PCs, subsequently leading to motor deficits and ASD-like behaviors in mice. These results provide novel insight into the pathophysiological mechanisms by which CSMD3 mutations cause impairments in cerebellar function that may contribute to ASD.

PDZK1 confers sensitivity to sunitinib in clear cell renal cell carcinoma by suppressing the PDGFR-ß pathway.

BACKGROUND: Sunitinib has emerged as the primary treatment for advanced or metastatic clear cell renal cell carcinoma (ccRCC) due to its significant improvement in patients' average survival time. However, drug resistance and adverse effects of sunitinib pose challenges to its clinical benefits. METHODS: The differentially expressed genes (DEGs) associated with sunitinib sensitivity and resistance in ccRCC were investigated. Cell counting kit-8, plate colony formation, flow cytometry and subcutaneous xenograft tumor model assays were employed to explore the effects of PDZK1 on ccRCC. Further research on the molecular mechanism was conducted through western blot, co-immunoprecipitation, immunofluorescence co-localization and immunohistochemical staining. RESULTS: We elucidated that PDZK1 is significantly downregulated in sunitinib-resistant ccRCC specimens, and PDZK1 negatively regulates the phosphorylation of PDGFR-ß and the activation of its downstream pathways through interaction with PDGFR-ß. The dysregulated low levels of PDZK1 contribute to inadequate inhibition of cell proliferation, tumor growth, and insensitivity to sunitinib treatment. Notably, our preclinical investigations showed that miR-15b antagomirs enhance sunitinib cytotoxic effects against ccRCC cells by upregulating PDZK1 levels, suggesting their potential in overcoming sunitinib resistance. CONCLUSIONS: Our findings establish the miR-15b/PDZK1/PDGFR-ß axis as a promising therapeutic target and a novel predictor for ccRCC patients' response to sunitinib treatment.

Nucleolar HEAT Repeat Containing 1 Up-regulated by the Mechanistic Target of Rapamycin Complex 1 Signaling Promotes Hepatocellular Carcinoma Growth by Dominating Ribosome Biogenesis and Proteome Homeostasis.

BACKGROUND & AIMS: Hyperactivation of ribosome biogenesis leads to hepatocyte transformation and plays pivotal roles in hepatocellular carcinoma (HCC) development. We aimed to identify critical ribosome biogenesis proteins that are overexpressed and crucial in HCC progression. METHODS: HEAT repeat containing 1 (HEATR1) expression and clinical correlations were analyzed using The Cancer Genome Atlas and Gene Expression Omnibus databases and further evaluated by immunohistochemical analysis of an HCC tissue microarray. Gene expression was knocked down by small interfering RNA. HEATR1-knockdown cells were subjected to viability, cell cycle, and apoptosis assays and used to establish subcutaneous and orthotopic tumor models. Chromatin immunoprecipitation and quantitative polymerase chain reaction were performed to detect the association of candidate proteins with specific DNA sequences. Endogenous coimmunoprecipitation combined with mass spectrometry was used to identify protein interactions. We performed immunoblot and immunofluorescence assays to detect and localize proteins in cells. The nucleolus ultrastructure was detected by transmission electron microscopy. Click-iT (Thermo Fisher Scientific) RNA imaging and puromycin incorporation assays were used to measure nascent ribosomal RNA and protein synthesis, respectively. Proteasome activity, 20S proteasome foci formation, and protein stability were evaluated in HEATR1-knockdown HCC cells. RESULTS: HEATR1 was the most up-regulated gene in a set of ribosome biogenesis mediators in HCC samples. High expression of HEATR1 was associated with poor survival and malignant clinicopathologic features in patients with HCC and contributed to HCC growth in vitro and in vivo. HEATR1 expression was regulated by the transcription factor specificity protein 1, which can be activated by insulin-like growth factor 1-mammalian target of rapamycin complex 1 signaling in HCC cells. HEATR1 localized predominantly in the nucleolus, bound to ribosomal DNA, and was associated with RNA polymerase I transcription/processing factors. Knockdown of HEATR1 disrupted ribosomal RNA biogenesis and impaired nascent protein synthesis, leading to reduced cytoplasmic proteasome activity and inhibitory-κB/nuclear factor-κB signaling. Moreover, HEATR1 knockdown induced nucleolar stress with increased nuclear proteasome activity and inactivation of the nucleophosmin 1-MYC axis. CONCLUSIONS: Our study revealed that HEATR1 is up-regulated by insulin-like growth factor 1-mammalian target of rapamycin complex 1-specificity protein 1 signaling in HCC and functions as a crucial regulator of ribosome biogenesis and proteome homeostasis to promote HCC development.

Valorization of Corn Straw for Production of Glucose by Two-step Depolymerization.

Depolymerization of the cellulose part in lignocellulose to glucose is a significant step for lignocellulose valorization. As one of the main by-products of agricultural biomass in crop-producing filed, valorization of corn straw has attracted considerable attention. In this study, a two-step depolymerizing strategy of high-pressure CO2-H2O pretreatment and oxidation-hydrolysis was applied for selective depolymerization of the cellulose component of corn straw to glucose production. Most part of the hemicellulose component could be removed through high-pressure CO2-H2O pretreatment in the presence of low concentration of acetic acid, and then as high as 32.2% yield of glucose was achieved in water at 170 oC for 6 h without additional catalyst. The active acid sites generated during the partial oxidation of hydroxymethyl groups to carboxyl groups on glucose units of cellulose was shown to be crucial for the efficient valorization of corn straw for glucose production.

Effects of positive end-expiratory pressure on regional cerebral oxygen saturation in elderly patients undergoing thoracic surgery during one-lung ventilation: a randomized crossover-controlled trial.

BACKGROUND: A significant reduction in regional cerebral oxygen saturation (rSO2) is commonly observed during one-lung ventilation (OLV), while positive end-expiratory pressure (PEEP) can improve oxygenation. We compared the effects of three different PEEP levels on rSO2, pulmonary oxygenation, and hemodynamics during OLV. METHODS: Forty-three elderly patients who underwent thoracoscopic lobectomy were randomly assigned to one of six PEEP combinations which used a crossover design of 3 levels of PEEP-0 cmH2O, 5 cmH2O, and 10 cmH2O. The primary endpoint was rSO2 in patients receiving OLV 20 min after adjusting the PEEP. The secondary outcomes included hemodynamic and respiratory variables. RESULTS: After exclusion, thirty-six patients (36.11% female; age range: 60-76 year) were assigned to six groups (n = 6 in each group). The rSO2 was highest at OLV(0) than at OLV(10) (difference, 2.889%; [95% CI, 0.573 to 5.204%]; p = 0.008). Arterial oxygen partial pressure (PaO2) was lowest at OLV(0) compared with OLV(5) (difference, -62.639 mmHg; [95% CI, -106.170 to -19.108 mmHg]; p = 0.005) or OLV(10) (difference, -73.389 mmHg; [95% CI, -117.852 to -28.925 mmHg]; p = 0.001), while peak airway pressure (Ppeak) was lower at OLV(0) (difference, -4.222 mmHg; [95% CI, -5.140 to -3.304 mmHg]; p < 0.001) and OLV(5) (difference, -3.139 mmHg; [95% CI, -4.110 to -2.167 mmHg]; p < 0.001) than at OLV(10). CONCLUSIONS: PEEP with 10 cmH2O makes rSO2 decrease compared with 0 cmH2O. Applying PEEP with 5 cmH2O during OLV in elderly patients can improve oxygenation and maintain high rSO2 levels, without significantly increasing peak airway pressure compared to not using PEEP. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2200060112 on 19 May 2022.

Chlorpyrifos degradation and its impacts on phosphorus bioavailability in microplastic-contaminated soil.

Pesticide residues and microplastics (MPs) in agricultural soils are two major concerns for soil health and food safety. The degradation of chlorpyrifos (CPF), an organophosphorus pesticide, releases phosphates. This process may be affected by the presence of MPs in the soil. The combination of CPF and MPs presence in the soil may thus produce interaction effects that alter the soil phosphorus (P) balance. This study explores the degradation pathways of CPF (6 mg kg-1, 12 mg kg-1 of CPF addition) in soils with different levels of polylactic acid MPs (PLA-MPs) (0.0 %, 0.1 %, 0.5 %, 1.0 % w/w), and analyzes soil P fractions and phosphatase enzyme activities to investigate soil P bioavailability under different treatments. Results show that the degradation of CPF fits to a first-order decay model, with half-lives (DT50) ranging from 11.0 to 14.8 d depending on PLA-MPs treatment. The concentration of its metabolite 3, 5, 6-trichloropyridine 2-phenol (TCP) reached a peak of 0.93-1.67 mg kg-1 within 7-14 days. Similarly, the degradation of CPF led to a significant transient increase in P bioavailability within 3-7 days (p < 0.05), with a peak range of 22.55-26.01 mg kg-1 for Olsen-P content and a peak range of 4.63-6.76 % for the proportions of available P fractions (H2O-P+NaHCO3-P+NaOH-P), before returning to prior levels (Olsen-P: 11.28-19.52 mg kg-1; available soil P fractions: 4.15-5.61 %). CPF degradation (6 mg kg-1) was significantly inhibited in soil with 1.0 % PLA-MPs addition. The effects of MPs and CPF on soil P fractions occur at different time frames, implying that their modes of action and interactions with soil microbes differ.

Vacuolar protein sorting 35 (VPS35) acts as a tumor promoter via facilitating cell cycle progression in pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is insidious and highly malignant with extremely poor prognosis and drug resistance to current chemotherapies. Therefore, there is a critical need to investigate the molecular mechanism underlying PDAC progression to develop promising diagnostic and therapeutic interventions. In parallel, vacuolar protein sorting (VPS) proteins, involved in the sorting, transportation, and localization of membrane proteins, have gradually attracted the attention of researchers in the development of cancers. Although VPS35 has been reported to promote carcinoma progression, the specific molecular mechanism is still unclear. Here, we determined the impact of VPS35 on the tumorigenesis of PDAC and explored the underlying molecular mechanism. We performed a pan-cancer analysis of 46 VPS genes using RNAseq data from GTEx (control) and TCGA (tumor) and predicted potential functions of VPS35 in PDAC by enrichment analysis. Furthermore, cell cloning experiments, gene knockout, cell cycle analysis, immunohistochemistry, and other molecular and biochemical experiments were used to validate the function of VPS35. Consequently, VPS35 was found overexpressed in multiple cancers and correlated with the poor prognosis of PDAC. Meanwhile, we verified that VPS35 could modulate the cell cycle and promote tumor cell growth in PDAC. Collectively, we provide solid evidence that VPS35 facilitates the cell cycle progression as a critical novel target in PDAC clinical therapy.

Effects of Microplastics on the Transport of Soil Dissolved Organic Matter in the Loess Plateau of China.

Microplastics (MPs) pollution and dissolved organic matter (DOM) affect soil quality and functions. However, the effect of MPs on DOM and underlying mechanisms have not been clarified, which poses a challenge to maintaining soil health. Under environmentally relevant conditions, we evaluated the major role of polypropylene particles at four micron-level sizes (20, 200, and 500 µm and mixed) in regulating changes in soil DOM content. We found that an increase in soil aeration by medium and high-intensity (>0.5%) MPs may reduce NH4+ leaching by accelerating soil nitrification. However, MPs have a positive effect on soil nutrient retention through the adsorption of PO43- (13.30-34.46%) and NH4+ (9.03-19.65%) and their leached dissolved organic carbon (MP-leached dissolved organic carbon, MP-DOC), thereby maintaining the dynamic balance of soil nutrients. The regulating ion (Ca2+) is also an important competitor in the MP-DOM adsorption system, and changes in its intensity are dynamically involved in the adsorption process. These findings can help predict the response of soil processes, especially nutrient cycling, to persistent anthropogenic stressors, improve risk management policies on MPs, and facilitate the protection of soil health and function, especially in future agricultural contexts.

The expression and clinical significance of STAMBP in breast cancer.

BACKGROUND: Breast cancer is the leading cause of death from cancer in women worldwide. STAMBP functions as a JAMM family deubiquitinating enzyme that modulates the stability of substrate proteins in cells by cleaving ubiquitin moieties. The expression of STAMBP and its clinical significance in breast cancer remain unclear. METHODS AND RESULTS: The level of the STAMBP protein in noncancerous and tumor tissues of breast cancer patients was examined by immunohistochemical staining. The expression of STAMBP mRNA in tissues based on healthy individual and breast cancer patient data in the TCGA database was evaluated. The association between the expression of STAMBP mRNA and clinical features and prognosis was evaluated using TCGA database. Cell growth was assessed by Cell Counting Kit-8 (CCK-8) assay, and cell migration and invasion were assessed by wound healing and Transwell assays. Activation of the ERK signaling was detected by Western blotting. The expression of STAMBP was markedly upregulated in the cytoplasm of tumor cells from breast cancer patients. The level of STAMBP was closely associated with the tumor subtype and size and the TNM stage of the breast cancer patients. Importantly, high expression of STAMBP predicted poor overall survival (OS) for breast cancer patients. Furthermore, knockdown of STAMBP expression reduced cell mobility and invasion of breast cancer cells. Notably, the phosphorylation of EGFR and ERK was markedly reduced in STAMBP-knockdown cells. CONCLUSION: STAMBP plays a critical role in the progression of breast cancer and may serve as a biomarker to monitor the progression of the disease.

The effect of capnography on the incidence of hypoxia during sedation for EGD and colonoscopy in mildly obese patients: a randomized, controlled study.

BACKGROUND: By continually monitoring end-tidal carbon dioxide concentrations, capnography can detect abnormal ventilation or apnoea early. This randomized, controlled study explored the effect of early intervention with capnography on the incidence of hypoxia in mildly obese patients undergoing sedation for esophagogastroduodenoscopy (EGD) and colonoscopy. METHODS: This is a single-center, randomized, single-blind, parallel-assignment, controlled trial. Mildly obese patients (28 kg/m2 ≤ BMI < 40 kg/m2) undergoing sedation for EGD and colonoscopy were randomly assigned to either the standard or capnography group. Standard cardiopulmonary monitoring equipment was used in both groups, and additional capnography was performed in the capnography group. In the event of inadequate alveolar ventilation during sedation, five interventions were administered in sequence (a-e) : a: increasing oxygen flow (5 L/min); b: a chin lift or jaw thrust maneuver; c: placement of the nasopharyngeal airway and chin lift; d: mask positive-pressure ventilation, and e: ventilator-assisted ventilation with tube insertion. The primary outcome was the incidence of hypoxia (SpO2 < 90%, ≥ 10 s) in each group. The secondary outcomes included the incidence of severe hypoxia (SpO2 ≤ 85%), subclinical respiratory depression (90% ≤ SpO2 < 95%), interventions, minimum SpO2 during operation, patient satisfaction, endoscopist satisfaction, and other adverse events of anesthesia sedation. RESULTS: 228 patients were included (capnography group = 112; standard group = 113; three patients were excluded) in this study. The incidence of hypoxia was significantly lower in the capnography group than in the standard group (13.4% vs. 30.1%, P = 0.002). Subclinical respiratory depression in the capnography group was higher than that of the standard group (30.4% vs. 17.7%, P = 0.026). There was only a 5.4% incidence of severe hypoxia in the capnography group compared with 14.2% in the standard group (P = 0.026). During sedation, 96 and 34 individuals in the capnography and standard groups, respectively, underwent the intervention. There was a statistically significant difference (P < 0.0001) in the number of the last intraoperative intervention between the two groups ( a:47 vs. 1, b:46 vs. 26, c:2 vs. 5, d:1 vs. 2, e:0 vs. 0 ). No significant differences were found between the two groups in terms of minimum SpO2 during operation, patient satisfaction, or endoscopist satisfaction rating. There was no statistically significant difference in adverse events of anesthesia sedation between the two groups. CONCLUSION: Capnography during sedation for EGD and colonoscopy allows for the detection of apnea and altered breathing patterns in mildly obese patients before SpO2 is reduced. Effective intervention measures are given to patients within this time frame, which reduces the incidence of hypoxia and severe hypoxia in patients. TRIAL REGISTRATION: Ethical approval was granted by the Medical Ethics Committee (Chairperson Professor Tian Hui) of Qilu Hospital, Shandong University ((Ke) Lun Audit 2021 (186)) on 15/07/2021. The study was registered ( https://www.chictr.org.cn ) on 23/10/2021(ChiCTR2100052234). Designed and reported using CONSORT statements.

Response of soybean and maize roots and soil enzyme activities to biodegradable microplastics contaminated soil.

Although biodegradable plastic film is a promising alternative product for reducing polyethylene plastic pollution in agricultural soils, the effects of its residues on plant growth and soil properties remain unclear. In this study, we conducted an experiment to investigate root properties and soil enzyme activities in Poly (butylene adipate-co-terephthalate) microplastics (PBAT-MPs) contaminated soil (0 % (CK), 0.1 %, 0.2 %, 0.5 % and 1 % of dry soil weight) with soybean (Glycine max (Linn.) Merr.) and maize (Zea mays L.). The results show that PBAT-MP accumulation in soil negatively affects root growth, and alter soil enzyme activities, which may then constrain C/N cycling and potential yields. For soybean, the total root length, total root surface area and root biomass decreased by 34 %- 58 %, 34 %- 54 % and 25 %- 40 % at the harvesting stage compared to CK, respectively. The negative effects of PBAT-MPs on maize roots were greater than on soybean roots. The total root length, root surface area and root biomass of maize decreased by 37 %- 71 %, 33 %- 71 % and 24 %- 64 % at the tasseling and harvesting stage, respectively (p < 0.05). Furthermore, a statistical analysis of the data indicates that the inhibition of soybean and maize root growth by PBAT-MP accumulation was mediated by the significantly different impacts of PBAT-MP addition on C-enzyme (ß-xylosidase, cellobiohydrolase, ß-glucosidase) and N-enzyme activities (leucine-aminopeptidase, N-acetyl-ß-glucosaminidase, alanine aminotransferase) in rhizosphere and non-rhizosphere soil, possibly due to interactions with plant-specific root exudates and microbial communities. These findings show the potential risks posed by biodegradable microplastics on the plant-soil system, and suggest that biodegradable plastic film should be applied with caution.

PEDOT Films Doped with Titanyl Oxalate as Chemiresistive and Colorimetric Dual-Mode Sensors for the Detection of Hydrogen Peroxide Vapor.

Hydrogen peroxide (H2O2) is commonly used as an oxidizing, bleaching, or antiseptic agent. It is also hazardous at increased concentrations. It is therefore crucial to monitor the presence and concentration of H2O2, particularly in the vapor phase. However, it remains a challenge for many state-of-the-art chemical sensors (e.g., metal oxides) to detect hydrogen peroxide vapor (HPV) because of the interference of moisture in the form of humidity. Moisture, in the form of humidity, is guaranteed to be present in HPV to some extent. To meet this challenge, herein, we report a novel composite material based on poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) doped with ammonium titanyl oxalate (ATO). This material can be fabricated as a thin film on electrode substrates for use in chemiresistive sensing of HPV. The adsorbed H2O2 will react with ATO, causing a colorimetric response in the material body. Combining colorimetric and chemiresistive responses resulted in a more reliable dual-function sensing method that improved the selectivity and sensitivity. Moreover, the composite film of PEDOT:PSS-ATO could be coated with a layer of pure PEDOT via in situ electrochemical synthesis. The pure PEDOT layer was hydrophobic, shielding the sensor material underneath from coming into contact with moisture. This was shown to mitigate the interference of humidity when detecting H2O2. A combination of these material properties makes the double-layer composite film, namely PEDOT:PSS-ATO/PEDOT, an ideal sensor platform for the detection of HPV. For example, upon a 9 min exposure to HPV at a concentration of 1.9 ppm, the electrical resistance of the film increased threefold, surpassing the bounds of the safety threshold. Meanwhile, the colorimetric response observed was 2.55 (defined as the color change ratio), a ratio at which the color change could be easily seen by the naked eye and quantified. We expect that this reported dual-mode sensor will find extensive practical applications in the fields of health and security with real-time, onsite monitoring of HPV.

Response of soil phosphatase activity and soil phosphorus fractions to the application of chloropicrin and azoxystrobin in ginger cultivation.

BACKGROUND: Soil fumigation can change soil nutrient cycling processes by affecting soil beneficial microorganisms, which is a key issue for soil fertility. However, the effect of combined application of fumigant and fungicide on soil phosphorus (P) availability remains largely unclear. We investigated the effects of the fumigant chloropicrin (CP) and the fungicide azoxystrobin (AZO) on soil phosphatase activity and soil P fractions in ginger production using a 28-week pot experiment with six treatments: control (CK), a single application of AZO (AZO1), double applications of AZO (AZO2), CP-fumigated soil without AZO (CP), CP combined with AZO1 (CP + AZO1) and CP combined with AZO2 (CP + AZO2). RESULTS: AZO application alone significantly increased the soil labile P fractions (Resin-P + NaHCO3 -Pi + NaOH-Pi) at 9 weeks after planting (WAP) but decreased the soil phosphatase activity at 28 WAP. CP fumigation significantly reduced the soil phosphatase activity but increased the proportions of soil labile P fractions (Resin-P + NaHCO3 -Pi + NaHCO3 -Po) to total P (TP) by 9.0-15.5% throughout the experiment. The combined application of CP and AZO had a synergistic effect on soil phosphatase activity and soil P fractions compared with a single application. CONCLUSION: Although AZO application and CP fumigation can increase soil available P in the short term, they might negatively affect soil fertility in the long run by inhibiting soil phosphatase activity. Soil microbial activities, especially microorganisms related to P cycling, may be responsible for the variations in soil P availability, but further research is needed. © 2023 Society of Chemical Industry.

Fetal Tethered Spinal Cord: Diagnostic Features and Its Association with Congenital Anomalies.

OBJECTIVE: We assessed the frequency and type of associated congenital anomalies encountered with fetal tethered spinal cord (TSC) determined prenatally. METHOD: A retrospective review was conducted based on the associated fetal abnormalities following diagnosis of low-lying fetal conus medullaris during the prenatal ultrasound. RESULTS: Of the 26 fetuses with low-lying conus medullaris, four were solitary TSC and 22 had TSC combined with associated congenital malformations, including four cases with spina bifida occulta, four cases with spina bifida aperta, one case with severe hydrocephalus, and 13 cases with multisystem congenital malformations. Among all the 13 cases with combined multisystem congenital malformations, four cases had vertebral defects, anal anomalies, cardiac defects, trachea-esophageal fistula, renal anomalies, and limb anomalies (VACTERL) syndrome, two cases had combined kidney development abnormalities, one case had cloacal exstrophy (OEIS syndrome), and six cases had chromosomal abnormalities (one case of chromosome 7q deletion, two cases of trisomy 13 syndrome, one case of trisomy 18 syndrome, one case of trisomy 9 syndrome, and one case of chromosome 4p deletion). CONCLUSIONS: Low-lying conus medullaris found during prenatal ultrasound examination were often associated with neural tube malformations or multi-systemic complex developmental malformations. The frequency of chromosomal abnormalities was 23.1%.

Contribution of AMPA Receptor-Mediated LTD in LA/BLA-CeA Pathway to Comorbid Aversive and Depressive Symptoms in Neuropathic Pain.

Comorbid anxiety and depressive symptoms in chronic pain are a common health problem, but the underlying mechanisms remain unclear. Previously, we have demonstrated that sensitization of the CeA neurons via decreased GABAergic inhibition contributes to anxiety-like behaviors in neuropathic pain rats. In this study, by using male Sprague Dawley rats, we reported that the CeA plays a key role in processing both sensory and negative emotional-affective components of neuropathic pain. Bilateral electrolytic lesions of CeA, but not lateral/basolateral nucleus of the amygdala (LA/BLA), abrogated both pain hypersensitivity and aversive and depressive symptoms of neuropathic rats induced by spinal nerve ligation (SNL). Moreover, SNL rats showed structural and functional neuroplasticity manifested as reduced dendritic spines on the CeA neurons and enhanced LTD at the LA/BLA-CeA synapse. Disruption of GluA2-containing AMPAR trafficking and endocytosis from synapses using synthetic peptides, either pep2-EVKI or Tat-GluA2(3Y), restored the enhanced LTD at the LA/BLA-CeA synapse, and alleviated the mechanical allodynia and comorbid aversive and depressive symptoms in neuropathic rats, indicating that the endocytosis of GluA2-containing AMPARs from synapses is probably involved in the LTD at the LA/BLA-CeA synapse and the comorbid aversive and depressive symptoms in neuropathic pain in SNL-operated rats. These data provide a novel mechanism for elucidating comorbid aversive and depressive symptoms in neuropathic pain and highlight that structural and functional neuroplasticity in the amygdala may be important as a promising therapeutic target for comorbid negative emotional-affective disorders in chronic pain.SIGNIFICANCE STATEMENT Several studies have demonstrated the high comorbidity of negative affective disorders in patients with chronic pain. Understanding the affective aspects related to chronic pain may facilitate the development of novel therapies for more effective management. Here, we unravel that the CeA plays a key role in processing both sensory and negative emotional-affective components of neuropathic pain, and LTD at the amygdaloid LA/BLA-CeA synapse mediated by GluA2-containing AMPAR endocytosis underlies the comorbid aversive and depressive symptoms in neuropathic pain. This study provides a novel mechanism for elucidating comorbid aversive and depressive symptoms in neuropathic pain and highlights that structural and functional neuroplasticity in the amygdala may be important as a promising therapeutic target for comorbid negative emotional-affective disorders in chronic pain.

E3 ubiquitin ligase MAGI3 degrades c-Myc and acts as a predictor for chemotherapy response in colorectal cancer.

BACKGROUND: Recurrence and chemoresistance constitute the leading cause of death in colorectal cancer (CRC). Thus, it is of great significance to clarify the underlying mechanisms and identify predictors for tailoring adjuvant chemotherapy to improve the outcome of CRC. METHODS: By screening differentially expressed genes (DEGs), constructing random forest classification and ranking the importance of DEGs, we identified membrane associated guanylate kinase, WW and PDZ domain containing 3 (MAGI3) as an important gene in CRC recurrence. Immunohistochemical and western blot assays were employed to further detect MAGI3 expression in CRC tissues and cell lines. Cell counting kit-8, plate colony formation, flow cytometry, sub-cutaneous injection and azoxymethane plus dextran sulfate sodium induced mice CRC assays were employed to explore the effects of MAGI3 on proliferation, growth, cell cycle, apoptosis, xenograft formation and chemotherapy resistance of CRC. The underlying molecular mechanisms were further investigated through gene set enrichment analysis, quantitative real-time PCR, western blot, co-immunoprecipitation, ubiquitination, GST fusion protein pull-down and immunohistochemical staining assays. RESULTS: Our results showed that dysregulated low level of MAGI3 was correlated with recurrence and poor prognosis of CRC. MAGI3 was identified as a novel substrate-binding subunit of SKP1-Cullin E3 ligase to recognize c-Myc, and process c-Myc ubiquitination and degradation. Expression of MAGI3 in CRC cells inhibited cell growth, promoted apoptosis and chemosensitivity to fluoropyrimidine-based chemotherapy by suppressing activation of c-Myc in vitro and in vivo. In clinic, the stage II/III CRC patients with MAGI3-high had a significantly good recurrence-free survival (~ 80%, 5-year), and were not necessary for further adjuvant chemotherapy. The patients with MAGI3-medium had a robustly good response rate or recurrence-free survival with fluoropyrimidine-based chemotherapy, and were recommended to undergo fluoropyrimidine-based adjuvant chemotherapy. CONCLUSIONS: MAGI3 is a novel E3 ubiquitin ligase by degradation of c-Myc to regulate CRC development and may act as a potential predictor of adjuvant chemotherapy for CRC patients.

Identification of two SPRY isoforms SPRY1 and SPRY3 by atomic force microscopy at the single-molecule level.

Growing reports indicate that Sprouty (SPRY) isoforms act as inhibitors or promoters in various types of cancers. And the occurrence of different cancers may be related to the abnormal expression of one of the SPRY isoforms. The identification of SPRY isoforms thus plays a particularly important role in determining which isoform's aberrant expression inhibits or promotes cancer. But their own properties, such as similarities in the structure and molecular weight, make their identification particularly difficult. In this article, we propose a novel method to identify SPRY isoforms using atomic force microscopy (AFM) by observing differential binding of different SPRY isoforms to bovine serum albumin (BSA), which can be used to distinguish SPRY isoforms at the single-molecule level. Specific binding of SPRY1 and BSA was observed by AFM. The reduction in the number of monomeric protein molecules caused by the partial depletion of these two proteins during binding is also consistent with the weakening of the monomeric protein bands in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). At the same time, the arrangement of the two proteins in a tightly bound complex was also observed. However, the SPRY3 isoform did not interact with BSA to cause aggregation, and the diameter and height of the two proteins did not change significantly compared to those before the reaction. In this way, with the participation of BSA, the two isoforms, SPRY1 and SPRY3, can be identified and separated using atomic force microscopy. In addition, the experimental result that the formation of the SPRY1-BSA complex can selectively reduce the concentration of SPRY1 isoforms in the environment will also contribute to future research on anticancer drugs influenced by SPRY1.

Effect of different polymers of microplastics on soil organic carbon and nitrogen - A mesocosm experiment.

Agricultural microplastic pollution has become a growing concern. Unfortunately, the impacts of microplastics (MPs) on agricultural soil carbon and nitrogen dynamics have not been sufficiently reported. In an attempt to remedy this, we conducted a 105-day out-door mesocosm experiment in a soil-plant system using sandy soils amended with two types of MPs, low-density polyethylene (LDPE-MPs) and biodegradable (Bio-MPs), at concentrations of 0.0% (control), 0.5%, 1.0%, 1.5%, 2.0% and 2.5% (w/w, weight ratio of microplastics to air-dry soil). Soil organic matter (SOM), dissolved organic carbon (DOC), permanganate oxidizable carbon (POXC), available nitrogen (AN) of N-NH4+ and N-NO3-, and dissolved organic nitrogen (DON) were measured on day 46 (D46) and 105 (D105) of the experiment. SOM was also measured after microplastics were mixed into soils (D0). For LDPE-MPs treatments, SOM on D0, D46 and D105 showed no significant differences, while for Bio-MPs treatments, SOM significantly (p < 0.05) decreased from D0 to D46. Compared to the control, soil POXC was significantly (p = 0.001) lowered by 0.5%, 1.0% and 2.5% LDPE-MPs and ≥ 1.0% Bio-MPs on D105. LDPE-MPs showed no significant effects on soil DOC and nitrogen cycling. 2.0% and 2.5% Bio-MPs showed significantly higher (p < 0.001) DOC and DON (on D46 and D105) and ≥1.5% Bio-MPs showed significantly lower (p = 0.02) AN (on D46). Overall, Bio-MPs exerted stronger effects on the dynamics of soil carbon and nitrogen cycling. In conclusion, microplastics might pose serious threats to agroecosystems and further research is needed.

Influence of Nutritional Status on Prognosis of Stroke Patients With Dysphagia.

Context: Stroke is an acute cerebrovascular disease and a neurological disorder that occurs due to a cerebral arterial embolism and rupture. Acute stroke is often accompanied by dysphagia, which reduces patients' intake of food and nutrients, decreases their nutritional status, and affects their quality of life. Objective: The study intended to identify the demographic and clinical characteristics of stroke patients with dysphagia and to explore the relationship of those characteristics to nutritional status and prognosis. Methods: The research team retrospectively collected the clinical data of patients to compare the nutritional status and prognoses of patients with different demographic and clinical characteristics. Setting: The study took place in the Department of Neurology at the First People's Hospital of Shenyang in Shenyang, China. Participants: Participants were 789 stroke patients with dysphagia who had been admitted to the general ward of the neurology departments of hospitals of Grade 3 or higher in Northeast China between January 2019 and September 2020. Based on the results of the Nutrition Risk Screening (NRS-2002) and Subjective Global Assessment (SGA) scales at baseline, participants were enrolled in this study. Outcome Measures: The outcomes were the correlations between participants' demographic and clinical characteristics and their nutritional statuses and prognoses. The Modified Rankin Scale (mRS) was used to evaluate the prognosis of the patients at seven days and three months after participants' enrollment in the study. Using the SPSS 26.0, a t test, chi-square test, and F test were performed to analyze and verify the presence of fundamental differences in baseline characteristics between participants with good nutrition and those with poor nutrition. Also, a statistical correlation analysis was performed. Results: The study showed that participants with different nutritional levels had statistically significant differences in the presence or absence of infections and body temperature and scores on the Standardized Swallowing Assessment (SSA) and National Institutes of Health Stroke Scale (NIHSS), with all P < .001. At baseline seven days after enrollment, the prognoses of participants were significantly different for different previous histories of stroke (P < .001), family history of stroke (P = .005), presence or absence of infections (P < .001), body temperature (P < .001), and SSA (P < .001) and NIHSS (P < .001) scale scores. At three months after enrollment, the prognoses of participants were significantly different for previous history of stroke (P = .003), different body temperatures (P < .001), presence or absence of infections(P < .001), and SSA (P < .001) and NIHSS (P < .001) scale scores. Age, gender, family history of stroke, smoking, alcohol consumption, previous history of stroke, education level, SSA scale score, NIHSS scale score, body mass index (BMI), body temperature, and infection were adjusted in the model. Nutritional status as classified by NRS-2002 and SGA was significantly correlated with prognosis (P < .001). The prognosis of stroke patients with dysphagia was associated with nutritional status by unconditional logistic regression. Conclusion: The prognosis of stroke patients with dysphagia is related to their nutritional status. A better nutritional status indicates the better prognosis, and vice versa. In clinical treatment, attention should be paid to use of a nutritional intervention.

Effects of chloropicrin fumigation and azoxystrobin application on ginger growth and phosphorus uptake.

Soil chloropicrin (CP) fumigation helps to increase crop yields by eliminating soil-borne diseases which inhibit plant growth. However, little is known about the effect of the CP fumigation combined with fungicide application on plant growth and nutrient uptake. In this study, we conducted a mesocosm experiment with six treatments: CK (untreated soil), AZO1 (a single application of azoxystrobin (AZO)), AZO2 (double applications of AZO), CP (CP fumigation with no AZO), CP+AZO1 (CP combined with AZO1) and CP+AZO2 (CP combined with AZO2) to investigate the effects of CP fumigation and AZO application on ginger growth and phosphorus (P) uptake. Results showed that a single application of AZO had no significant effect on ginger height, biomass and P uptake whether treated with or without CP fumigation, whereas double applications of AZO combined with CP fumigation significantly improved ginger height and the total amount of P in root (P < 0.05). Meanwhile, AZO residues were similar in all treatments with the same number of applications, with less than 50% remaining in the soil after 7 days applied, indicating that CP fumigation treatment did not influence AZO degradation in ginger cultivation. In addition, although the differences in P use efficiency observed across the different treatments were not significant, they nevertheless suggest that the P budget and soil microbial activity may contribute to those differences. Therefore, further studies should be done to link P cycling with microbial communities, and how these related to fumigation and fungicide application.