A distinct isoform of Msp300/Nesprin organizes the perinuclear microtubule organizing center in adipose cells.

In many cell types, disparate non-centrosomal microtubule-organizing centers (ncMTOCs) replace functional centrosomes and serve the unique needs of the cell types in which they are formed. In Drosophila fat body cells (adipocytes), an ncMTOC is organized on the nuclear surface. This perinuclear ncMTOC is anchored by Msp300, encoded by one of two Nesprin-encoding genes in Drosophila. Msp300 and the spectraplakin short stop (shot) are co-dependent for localization to the nuclear envelope to generate the ncMTOC where they recruit the microtubule minus-end stabilizer Patronin (CAMSAP). The fat body perinuclear ncMTOC requires Patronin, Ninein, and Msps (ortholog of ch-TOG), but does not require γ-tubulin for MT assembly. The Msp300 gene is complex, encoding at least eleven isoforms. Here we show that two Msp300 isoforms, Msp300-PE and -PG, are required and only one, Msp300-PE, appears sufficient for generation of the ncMTOC. Loss of Msp300-PE,-PG retains the presence of the other isoforms at the nuclear surface, indicating that they are not sufficient to generate the ncMTOC. Loss of Msp300-PE,-PG results in severe loss of localization of shot and Patronin, and disruption of the MT array. This results in nuclear mispositioning and loss of endosomal trafficking. Msp300-PE has an unusual domain structure including a lack of a KASH domain and very few spectrin repeats and appears therefore to have a highly derived function suited to generating an ncMTOC on the nuclear surface.

Ninein domains required for its localization, association with partners dynein and ensconsin, and microtubule organization.

Ninein (Nin) is a microtubule (MT) anchor at the subdistal appendages of mother centrioles and the pericentriolar material (PCM) of centrosomes that also functions to organize microtubules at non-centrosomal microtubule-organizing centers (ncMTOCs). In humans, the NIN gene is mutated in Seckel syndrome, an inherited developmental disorder. Here we dissect the protein domains involved in Nin's localization and interactions with dynein and ensconsin (ens/MAP7) and show that the association with ens cooperatively regulates microtubule assembly in Drosophila fat body cells. We define domains of Nin responsible for its localization to the ncMTOC on the fat body cell nuclear surface, localization within the nucleus, and association with Dynein light intermediate chain (Dlic) and ens, respectively. We show that Nin's association with ens synergistically regulates MT assembly. Together, these findings reveal novel features of Nin function and its regulation of a ncMTOC.

Influence Factors of the Therapeutic Effect of Budesonide Combined with N-Acetylcysteine in Children with Mycoplasma Pneumoniae Infection Analyzed by Lasso-Logistic and Construction of a Nomogram Prediction Model.

Objective: This study aims to analyze the factors influencing the efficacy of budesonide (BUD) combined with N-acetylcysteine (NAC) treatment in children with Mycoplasma pneumoniae (MP) infection through Lasso-Logistic analysis, construct a Nomogram predictive model, and provide personalized treatment plans for clinicians. Additionally, it aims to fill the knowledge gap regarding the treatment of MP-infected children with BUD combined with NAC. Methods: We retrospectively analyzed clinical data from 96 children treated with BUD and NAC for MP infection at our hospital from January 2022 to May 2023. Treatment outcomes were categorized as good or poor. Logistic regression and Lasso-Logistic analysis were used to identify independent factors influencing outcomes and construct a predictive Nomogram model, which was validated through ROC curve analysis. Results: Logistic regression identified prolonged fever (≥7 days), high fever, and elevated levels of TNF-α, IL-6, and CRP as independent risk factors for poor outcomes. The Nomogram model, based on these factors, demonstrated excellent predictive accuracy with a C-index of 0.992 and AUC values of 0.987 and 0.948 in the modeling and validation cohorts, respectively. Conclusion: The developed Nomogram model provides clinicians with a reliable tool to predict poor treatment outcomes in children with MP infection treated with BUD and NAC, supporting more personalized and effective treatment plans.

Clinical efficacy of budesonide combined with acetylcysteine in the treatment of mycoplasma pneumonia infection.

OBJECTIVE: Mycoplasma pneumoniae pneumonia (MPP) is a common respiratory tract infectious disease in children. The study aimed to elucidate the therapeutic efficacy of aerosolized budesonide and N-acetylcysteine combination therapy for MP infection in children. METHODS: One hundred and twenty children with MP infection were included and divided into the control group (received aerosol inhalation of budesonide) and the experimental group (aerosolized budesonide and N-acetylcysteine). After treatment, the disappearance time of clinical symptoms and efficacy were contrasted between the two groups. RESULTS: With the passage of treatment time, the children's cough score of the two groups were gradually reduced. The children in the experimental group got well from the cough faster than the control group, and the difference reached a significant level on the 5th and 7th days. The time required for fever, rale, and cough to disappear in the experimental group was shorter than those in the control group. As the treatment progressed, a gradual decrease in serum interleukin-6, tumor necrosis factor-α, and C-reactive protein values was detected in both groups, and the decrease was more significant in the experimental group. The total effective rate of the experimental group was 98.33%, which surpassed the control group (93.33%). CONCLUSION: Budesonide and N-acetylcysteine combination therapy in the treatment of MP infection in children has a significant effect, and can quickly relieve the clinical symptoms of children with good safety. It is worthy of widespread clinical use.

Analyzing the epidemic situation and influencing factors of childhood pneumonia during the COVID-19 epidemic period in Laiwu, China.

This study aimed to analyze the trends of childhood pneumonia before and after the corona virus disease 2019 (COVID-19) pandemic in Laiwu, China, and explore the associated risk factors to provide a reference for the prevention, control, and treatment of childhood pneumonia. In this cross-sectional study, data were collected from 11,716 children aged 3 to 12 years who underwent chest radiography/computed tomography from January 2018 to December 2021 in Laiwu, China. The generalized estimating equation model was used to analyze the time trend of the pneumonia detection rate. Multivariate logistic regression analysis was used to analyze the risk factors for pneumonia in children. The overall pneumonia detection rate was 40.3% (4721/11,716). The pneumonia detection rate was 41.5% and 39.5% in 2018 and 2019, respectively, before the COVID-19 pandemic, which decreased to 38.1% in 2020 during the pandemic and increased to 40.7% in 2021 after the pandemic. Multivariate logistic regression analysis showed that preterm birth (odds ratio [OR] = 1.68), frequent exposure to secondhand smoke (OR = 1.79), hospitalization ≥ 1 time in half a year (OR = 2.13), and history of allergic rhinitis (OR = 2.14) were risk factors for pneumonia in children. Wearing masks when outdoors (OR = 0.89) and engaging in regular physical activity (OR = 0.65) were protective factors. The pneumonia detection rate in children showed a decreasing trend during the COVID-19 pandemic in 2020 but an increasing trend after the pandemic in 2021. Premature birth, exposure to secondhand smoke, frequent hospitalization, and allergic rhinitis were risk factors for pneumonia in children. Wearing masks when outdoors and exercising may reduce the risk of pneumonia in children.

[Effects of Chinese Milk Vetch Returning Incorporated with Chemical Fertilizer Reduction on the Composition and Function of Soil Bacterial Communities in Paddy Fields].

Chinese milk vetch (Astragalus sinicus L.) is an important organic nutrient resource in the southern Henan rice-growing area. Thus, the effects of Chinese milk vetch (MV) returning incorporated with reduced chemical fertilizer on the physicochemical properties and bacterial community characteristics in paddy soil were studied. These results can provide a certain theoretical basis for the improvement of soil fertility and reduction of chemical fertilizer in this area. A field experiment was conducted for 12 consecutive years, involving six fertilization treatments (blank control, CK; 100% chemical fertilizer, F100; 80% chemical fertilizer+22.5 t·hm-2 MV, MV1F80; 80% chemical fertilizer+45 t·hm-2 MV, MV2F80; 60% chemical fertilizer+22.5 t·hm-2 MV, MV1F60; and 60% chemical fertilizer+45 t·hm-2 MV, MV2F60). The high-throughput sequencing method was used to compare the effects of different fertilization treatments on soil bacterial community diversity, composition, and structural characteristics. The FAPROTAX function prediction method was used to analyze the abundance differences of functional groups between different fertilization treatments. Additionally, combined with soil physicochemical properties and bacterial community characteristics, we explored the key soil environmental factors that changed the structure and functional characteristics of the soil bacterial community. Compared with that under CK, the soil bulk density (BD) under the MV returning incorporated with reduced chemical fertilizer treatment was decreased, whereas soil organic carbon (SOC), total nitrogen (TN), total phosphorus (TP), and total potassium (TK) were increased by 12.7%-35.5%, 38.2%-65.7%, 66.7%-95.2%, and 20.3%-31.6%, respectively. Compared with that under the F100 treatment, the Sobs index and Shannon diversity index of the bacterial community under the MV returning incorporated with reduced chemical fertilizer were decreased, and the Sobs index and Shannon diversity index were significantly positively correlated with BD (P<0.05) but significantly negatively correlated with SOC and TN (P<0.05). Compared with that under the F100 treatment, the relative abundances of Firmicutes under the MV1F80 and MV2F60 treatments were significantly increased by 82.2% and 67.4% (P<0.05), but the relative abundances of Acidobacteria were significantly reduced by 32.6% and 40.5% (P<0.05), respectively. The relative abundance of Actinobacteria under the MV2F60 treatment was significantly increased by 30.0% (P<0.05) compared with that under the F100 treatment. According to RDA analysis, soil SOC, TN, and TK were the main soil environmental factors that significantly affected bacterial community (P<0.05). Compared with that under CK and the F100 treatment, the abundance of functional groups of chemoheterotrophy, nitrogen fixation, fermentation, and ureolysis under the MV returning incorporated with reduced chemical fertilizer treatment were improved, whereas the abundance of functional groups of animal parasites or symbionts, all human pathogens, and human pathogen pneumonia were reduced, particularly under MV1F80 and MV2F60. To summarize, the long-term MV returning to the field incorporated with reduced chemical fertilizer improved the soil physical and chemical properties, thus changing the structure and functional characteristics of the soil bacterial communities, contributing to the improvement in the soil fertility, stability, and health of micro-ecosystems in paddy fields, thus ensuring the green and sustainable development of regional agriculture.

Study on the relationships between the accumulation and translocation of dry matter and nitrogen and flower/pod development into seeds and seed yields in Chinese milk vetch.

Further improvements to the yield potential of Chinese milk vetch seed are essential for the planting demand of green manure. Flower and pod development directly determines the number of seeds and the seed yield of Chinese milk vetch. However, the accumulation and translocation of dry matter and nitrogen between plant organs directly affects flower and pod development and morphological formation. There are few studies that analyse the relationship between the accumulation and transport of dry matter and nitrogen and the number of flowers, pods, grains and seed yield during Chinese milk vetch's critical development period. This study aimed to determine the seed yield response to dry matter and nitrogen accumulation and translocation during the Chinese milk vetch growth period and to quantify the relationship between these factors to predict Chinese milk vetch seed yield. Experiments were performed during the 2017-2018 and 2018-2019 growing seasons at the Dayuzhuang experimental field. The first experiment involved five foliar application stages (late wintering stage, returning green stage, squaring stage, pre-flowering stage, and 5 days after flowering) and six foliar application concentrations of borate solution (0, 500, 1000, 2000, 4000, and 6000 mg L-1). Experiment 2 included five foliar application stages (late wintering stage, returning green stage, squaring stage, pre-flowering stage, and 5 days after flowering) and six foliar application concentrations of paclobutrazol (0, 200, 300, 400, 500, and 600 mg L-1). When the dry matter mass in the full flowering stage was 3500-4500 kg hm-2, the seed yield reached more than 800 kg hm-2. When the translocated assimilates were stored in the vegetative organs before flowering, the assimilate translocation rate and their contributions to seed yield were 1500-1800 kg hm-2, 30-35%, and 28-38%, respectively, and the Chinese milk vetch seed yield was predicted to reach 800-1000 kg hm-2 at maturity. When the nitrogen translocation amount in the vegetative organs before flowering, the nitrogen translocation rate, and the contribution rate to the seed yield were 68-78 kg hm-2, 65-75%, and 75-85%, respectively, the Chinese milk vetch seed yield was predicted to reach 800-1000 kg hm-2 at maturity. If the accumulation and translocation index values of dry matter and nitrogen were lower or higher than the above ranges, the seed yield was lower than 800 kg hm-2. The results of this study revealed the mechanism by which dry matter and nitrogen accumulation and translocation affect the Chinese milk vetch seed yield. These findings enrich the seed yield formation theory of Chinese milk vetch. They provide an early determination and quantitative regulation of high and stable seed yield for Chinese milk vetch in the field and aid researchers to integrate multiple production technologies for the sustainable production of Chinese milk vetch.

Intraspinal extradural gout tophus in the lumbar vertebral canal: Case reports.

RATIONALE: Intraspinal gout tophus in the lumbar vertebral canal associated with gouty arthritis is rare. We present 2 cases with the first manifestations of a sequestrated intervertebral disc and an extradural tumor, and histopathologically proven to be gouty deposits in the lumbar vertebral canal. PATIENT CONCERNS: The 2 patients presented with typical radiculopathy symptoms and a positive straight leg raise. In 1 case, there was weakness of the left toe extensors, with a positive left femoral nerve traction test. Additionally, the left patellar tendon reflex was weak. In the other patient who was unable to walk, there was a sensory deficit in the saddle distribution. DIAGNOSIS: Histopathological examination of the specimens taken from the operation confirmed the presence of gouty deposits. INTERVENTIONS: Posterior decompression was performed in these 2 cases, and chalky-white materials were identified in the lumbar vertebral canal. OUTCOMES: No evidence of neoplasm, infection, or synovial cyst was found. LESSONS: Definitive diagnosis of intraspinal extradural gout tophus, mimicking a sequestrated intervertebral disc or an extradural tumor, may be difficult. The initial suspicion of intraspinal gouty deposits, based on the diagnostic/management algorithm, may effectively avoid incorrect diagnosis via a less invasive procedure than explorative laminectomy.

Coordination of Zika Virus Infection and Viroplasm Organization by Microtubules and Microtubule-Organizing Centers.

Zika virus (ZIKV) became a global health concern in 2016 due to its links to congenital microcephaly and other birth defects. Flaviviruses, including ZIKV, reorganize the endoplasmic reticulum (ER) to form a viroplasm, a compartment where virus particles are assembled. Microtubules (MTs) and microtubule-organizing centers (MTOCs) coordinate structural and trafficking functions in the cell, and MTs also support replication of flaviviruses. Here we investigated the roles of MTs and the cell's MTOCs on ZIKV viroplasm organization and virus production. We show that a toroidal-shaped viroplasm forms upon ZIKV infection, and MTs are organized at the viroplasm core and surrounding the viroplasm. We show that MTs are necessary for viroplasm organization and impact infectious virus production. In addition, the centrosome and the Golgi MTOC are closely associated with the viroplasm, and the centrosome coordinates the organization of the ZIKV viroplasm toroidal structure. Surprisingly, viroplasm formation and virus production are not significantly impaired when infected cells have no centrosomes and impaired Golgi MTOC, and we show that MTs are anchored to the viroplasm surface in these cells. We propose that the viroplasm is a site of MT organization, and the MTs organized at the viroplasm are sufficient for efficient virus production.

Flower and pod development, grain-setting characteristics and grain yield in Chinese milk vetch (Astragalus sinicus L.) in response to pre-anthesis foliar application of paclobutrazol.

The number of grains per unit land area is the most important grain yield component in Chinese milk vetch. Flower and pod survival seem to be critical determinants of grain number, which is related to the number of fertile flowers and pods during the anthesis period. Flower and pod growth are frequently considered the key determinants to establish grain number. The objective of this study was to explore the influences of paclobutrazol on flower and pod development, grain-setting characteristics and grain yield in Chinese milk vetch under different concentrations of foliar spray and try to explore the physiological regulatory mechanisms. Field experiments were carried out during the 2017-2018 and 2018-2019 growing seasons at the Dayuzhuang experimental field. The experiment involved the Chinese milk vetch cultivar "Xinzi No. 1" and six levels of foliar application of paclobutrazol, 0, 200, 300, 400, 500, and 600 mg L-1, in treatments CK, T1, T2, T3, T4, and T5, respectively. Foliar spray was applied once, at the squaring stage. In comparison with the CK treatment, all of the paclobutrazol treatments yielded, to various degrees, increased values of the number of inflorescences per unit area, number of pods per unit area, grain-setting rate of pods, and number of grains per pod in all six inflorescence layers, with the largest increases observed in the T3 treatment. In the T3 treatment compared with the CK treatment, from the first to sixth inflorescence layers, the number of inflorescences per unit area was increased by 34.07-58.97%, the number of pods per unit area was increased by 39.69-68.35%, the grain number per pod was increased by 44.31-53.69%, and the grain-setting rate of pods was increased by 1.84-4.89%. An analysis of yield composition revealed that the paclobutrazol spray treatment had little impact on the grain weight of Chinese milk vetch. The correlations between the concentration of paclobutrazol spray and the grain yield of Chinese milk vetch reached a significant level. Grain yield was highest at the paclobutrazol concentration of 373.10 mg/L. The inflorescence contents of gibberellic acid 3 (GA3), indole-3-acetic acid (IAA), and abscisic acid (ABA) were reduced, whereas that of cytokinin (CTK) was increased, by foliar application of paclobutrazol (400 mg L-1, T3 treatment) relative to CK treatment during the stages of flowers and pods developing into grains.

The Enigma of Centriole Loss in the 1182-4 Cell Line.

The Drosophilamelanogaster cell line 1182-4, which constitutively lacks centrioles, was established many years ago from haploid embryos laid by females homozygous for the maternal haploid (mh) mutation. This was the first clear example of animal cells regularly dividing in the absence of this organelle. However, the cause of the acentriolar nature of the 1182-4 cell line remained unclear and could not be clearly assigned to a particular genetic event. Here, we detail historically the longstanding mystery of the lack of centrioles in this Drosophila cell line. Recent advances, such as the characterization of the mh gene and the genomic analysis of 1182-4 cells, allow now a better understanding of the physiology of these cells. By combining these new data, we propose three reasonable hypotheses of the genesis of this remarkable phenotype.

Osteopontin exacerbates the progression of experimental autoimmune myasthenia gravis by affecting the differentiation of T cell subsets.

Osteopontin (OPN) is a multifunctional extracellular matrix phosphoprotein that has a specific and complicated structure, and contributes to numerous physiological and pathological activities. The mechanism of OPN in many diseases has been confirmed; however, the role of OPN in myasthenia gravis (MG) remains unclear. In this study, we recombined rat OPN protein in vitro, and assessed how OPN affects the development of autoimmunity using an experimental autoimmune myasthenia gravis (EAMG) rat model. The results showed that the concentration of OPN in serum was up-regulated. Both mRNA and protein levels in splenocytes increased in the EAMG model. OPN treatment in vitro strongly promoted the differentiation of Th1 cells, and inhibited the differentiation of Treg cells. Intraperitoneal injection of OPN revealed the early incidence of EAMG, and more serious disease. This effect was accompanied by an increased percentage of Th1 cells. In conclusion, OPN likely exacerbates the pathogenesis of EAMG by promoting the differentiation of Th1 cells and inhibiting the differentiation of Treg cells.

A perinuclear microtubule-organizing centre controls nuclear positioning and basement membrane secretion.

Non-centrosomal microtubule-organizing centres (ncMTOCs) have a variety of roles that are presumed to serve the diverse functions of the range of cell types in which they are found. ncMTOCs are diverse in their composition, subcellular localization and function. Here we report a perinuclear MTOC in Drosophila fat body cells that is anchored by the Nesprin homologue Msp300 at the cytoplasmic surface of the nucleus. Msp300 recruits the microtubule minus-end protein Patronin, a calmodulin-regulated spectrin-associated protein (CAMSAP) homologue, which functions redundantly with Ninein to further recruit the microtubule polymerase Msps-a member of the XMAP215 family-to assemble non-centrosomal microtubules and does so independently of the widespread microtubule nucleation factor γ-Tubulin. Functionally, the fat body ncMTOC and the radial microtubule arrays that it organizes are essential for nuclear positioning and for secretion of basement membrane components via retrograde dynein-dependent endosomal trafficking that restricts plasma membrane growth. Together, this study identifies a perinuclear ncMTOC with unique architecture that regulates microtubules, serving vital functions.

Wheat Grain Yield Increase in Response to Pre-Anthesis Foliar Application of 6-Benzylaminopurine Is Dependent on Floret Development.

Wheat yield is largely determined during the period prior to flowering, when the final numbers of fertile florets and grains per spike are established. The aim of this study was to assess the dynamics of floret primordia development in winter wheat in response to pre-anthesis application of a synthetic cytokinin, 6-benzylaminopurine (6-BA). We conducted an experiment in which two foliar spray treatments were applied (water or 6-BA) to Chinese winter wheat at 25 days after jointing during two growing seasons (2012-2013 and 2013-2014). Both the final grain number per spike and grain yield at maturity exhibited remarkable increases in response to the 6-BA treatment. Application of 6-BA increased the number of fertile florets in basal spikelets and, to a greater extent, in central spikelets. The mechanism by which 6-BA application affected the final number of fertile florets primarily involved suppression of the floret abortion rates. Application of 6-BA considerably reduced the abortion rates of basal, central and apical spikelet florets (by as much as 77% compared with the control), as well as the degeneration rates of basal and central spikelet florets, albeit to a lesser degree. The effect of 6-BA application on the likelihood of proximal florets being set was limited to the distal florets in the whole spike, whereas obvious increases in the likelihood of grain set under 6-BA treatment were observed in distal florets, primarily in central spikelet positions. The results of this study provide important evidence that 6-BA application to florets (final fertile floret production) results in an increased grain yield.

Lycorine: a potential broad-spectrum agent against crop pathogenic fungi.

A screening test showed that lycorine exhibited significant antifungal activity against 24 pathogenic crop fungi at concentrations of 500 µg/ml and 100 µg/ml, respectively. Fusarium graminearum was selected for antifungal mechanism studies by observing its mycelial morphology and investigating the variations in its conductivity. In addition, the substance absorption and metabolism of F. graminearum were explored. The mechanism was revealed as being one by which lycorine destroyed the cellular membrane and further influenced substance absorption and cell metabolism.