Molecular characterization, expression analysis and function identification of Pf_IL-12p35, Pf_IL-23p19 and Pf_IL-12p40 genes in yellow catfish (Pelteobagrus fulvidraco).

The interleukin-12 (IL-12) family is a class of heterodimeric cytokines that play crucial roles in pro-inflammatory and pro-stimulatory responses. Although some IL-12 and IL-23 paralogues have been found in fish, their functional activity in fish remains poorly understood. In this study, Pf_IL-12p35a/b, Pf_IL-23p19 and Pf_IL-12p40a/b/c genes were cloned from yellow catfish (Pelteobagrus fulvidraco), four α-helices were found in Pf_IL-12p35a/b and Pf_IL-23p19. The transcripts of these six genes were relatively high in mucus and immune tissues of healthy individuals, and in gill leukocytes. Following Edwardsiella ictaluri infection, Pf_IL-12p35a/b and Pf_IL-23p19 mRNAs were induced in brain and kidney (or head kidney), Pf_IL-12p40a mRNA was induced in gill, and Pf_IL-12p40b/c mRNAs were induced in brain and liver (or skin). The mRNA expression of these genes in PBLs was induced by phytohaemagglutinin (PHA) and polyinosinic-polycytidylic acid (poly I:C), while lipopolysaccharides (LPS) induced the mRNA expression of Pf_IL-12p35a and Pf_IL-12p40b/c in PBLs. After stimulation with recombinant (r) Pf_IL-12 and rPf_IL-23 subunit proteins, either alone or in combination, mRNA expression patterns of genes related to T helper cell development exhibited distinct differences. The results suggest that Pf_IL-12 and Pf_IL-23 subunits may play important roles in regulating immune responses to pathogens and T helper cell development.

Inhibition of TAF1B impairs ribosome biosynthesis and suppresses cell proliferation in stomach adenocarcinoma through promoting c-MYC mRNA degradation.

Hyperactivation of ribosome biosynthesis (RiBi) is a hallmark of cancer, and targeting ribosome biogenesis has emerged as a potential therapeutic strategy. The depletion of TAF1B, a major component of selectivity factor 1 (SL1), disrupts the pre-initiation complex, preventing RNA polymerase I from binding ribosomal DNA and inhibiting the hyperactivation of RiBi. Here, we investigate the role of TAF1B, in regulating RiBi and proliferation in stomach adenocarcinoma (STAD). We disclosed that the overexpression of TAF1B correlates with poor prognosis in STAD, and found that knocking down TAF1B effectively inhibits STAD cell proliferation and survival in vitro and in vivo. TAF1B knockdown may also induce nucleolar stress, and promote c-MYC degradation in STAD cells. Furthermore, we demonstrate that TAF1B depletion impairs rRNA gene transcription and processing, leading to reduced ribosome biogenesis. Collectively, our findings suggest that TAF1B may serve as a potential therapeutic target for STAD and highlight the importance of RiBi in cancer progression.

Recent progress, perspectives, and issues of engineered PD-L1 regulation nano-system to better cure tumor: A review.

Currently, immune checkpoint blockade (ICB) therapies that target the programmed cell death ligand-1 (PD-L1) have been used as revolutionary cancer treatments in the clinic. Apart from restoring the antitumor response of cytotoxic T cells by blocking the interaction between PD-L1 on tumor cells and programmed cell death-1 (PD-1) on T cells, PD-L1 proteins were also newly revealed to possess the capacity to accelerate DNA damage repair (DDR) and enhance tumor growth through multiple mechanisms, leading to the impaired efficacy of tumor therapies. Nevertheless, current free anti-PD-1/PD-L1 therapy still suffered from poor therapeutic outcomes in most solid tumors due to the non-selective tumor accumulation, ineludible severe cytotoxic effects, as well as the common occurrence of immune resistance. Recently, nanoparticles with efficient tumor-targeting capacity, tumor-responsive prosperity, and versatility for combination therapy were identified as new avenues for PD-L1 targeting cancer immunotherapies. In this review, we first summarized the multiple functions of PD-L1 protein in promoting tumor growth, accelerating DDR, as well as depressing immunotherapy efficacy. Following this, the effects and mechanisms of current clinically widespread tumor therapies on tumor PD-L1 expression were discussed. Then, we reviewed the recent advances in nanoparticles for anti-PD-L1 therapy via using PD-L1 antibodies, small interfering RNA (siRNA), microRNA (miRNA), clustered, regularly interspaced, short palindromic repeats (CRISPR), peptide, and small molecular drugs. At last, we discussed the challenges and perspectives to promote the clinical application of nanoparticles-based PD-L1-targeting therapy.

Conformable electrochemical devices for closed-loop wound management.

Chronic wounds arising from accidents, surgeries, or diseases impose a significant clinical and economic burden, underscoring the need for effective solutions to prevent severe complications. Recent advancements in materials science and electrochemical technology have facilitated the development of conformable electrochemical platforms for detection and management, incorporating monitoring, diagnosis, and treatment. Nevertheless, current wound detection and therapy systems face challenges related to the stability and specificity of sensor monitoring, as well as the need for on-site and comprehensive evaluation criteria to offer timely treatment guidance and follow-up care. This review provides a comprehensive overview of the closed-loop management system, emphasizing wound biomarker detection, wound assessment, and on-demand treatment, ultimately culminating in an integrated wound management approach by conformable electrochemical devices. Additionally, we explore the challenges, opportunities, and future prospects of soft and stretchable electrochemical biosensors, with the aim of enhancing the efficiency and timeliness of wound management.

Long-term effects of the COVID-19 pandemic on five mental and psychological disorders: in terms of the number of disease visits, drug consumption, and scale scores.

BACKGROUND: COVID-19 caused mild to severe infections in humans. The long-term epidemic environment harms people's mental health. To explore the impact of the epidemic on people's mental and psychological conditions, we surveyed in Wenzhou. METHODS: We collected the data of people who visited the First Affiliated Hospital of Wenzhou Medical University for five types of mental and psychological diseases from January 2018 to December 2021. Then, taking December 2019 as the cut-off point, the 48-month data were divided into the pre-epidemic group and the dur-epidemic group. Based on the above data, statistical analysis was done. RESULTS: From 2018 to 2021, the number of initial diagnoses, the number of disease visits, and drug consumption for these five types of mental and psychological diseases were all on the rise. Compared with the number of disease visits for all disorders in both psychiatry and neurology departments, it was found that the growth rate of these five diseases was higher than the growth rate of all disorders. We found that the number of disease visits, drug consumption, and scale scores after the COVID-19 outbreak were significantly different from those before the outbreak (P < 0.05). And the number of disease visits positively correlated with drug consumption (P < 0.0001, r = 0.9503), which verified the stability of the data. CONCLUSION: The epidemic environment has had a long-term and negative impact on people's mental and psychological conditions. Therefore, whether or not the epidemic is receding, we still need to be concerned about the impact of COVID-19 on mental and psychological health.

Calf thymus ds-DNA intercalation with pendimethalin herbicide at the surface of ZIF-8/Co/rGO/C3N4/ds-DNA/SPCE; A bio-sensing approach for pendimethalin quantification confirmed by molecular docking study.

Pendimethalin (PND) is a herbicide that is regarded to be possibly carcinogenic to humans and toxic to the environment. Herein, we fabricated a highly sensitive DNA biosensor based on ZIF-8/Co/rGO/C3N4 nanohybrid modification of a screen-printed carbon electrode (SPCE) to monitor PND in real samples. The layer-by-layer fabrication pathway was conducted to construct ZIF-8/Co/rGO/C3N4/ds-DNA/SPCE biosensor. The physicochemical characterization techniques confirmed the successful synthesis of ZIF-8/Co/rGO/C3N4 hybrid nanocomposite, as well as the appropriate modification of the SPCE surface. The utilization of ZIF-8/Co/rGO/C3N4 nanohybrid as a modifier was analyzed using. The electrochemical impedance spectroscopy results showed that the modified SPCE exhibited significantly lowered charge transfer resistance due to the enhancement of its electrical conductivity and facilitation of the transfer of charged particles. The proposed biosensor successfully quantified PND in a wide concentration range of 0.01-35 µM, with a limit of detection (LOD) value of 8.0 nM. The PND monitoring capability of the fabricated biosensor in real samples including rice, wheat, tap, and river water samples was verified with a recovery range of 98.2-105.6%. Moreover, to predict the interaction sites of PND herbicide with DNA, the molecular docking study was performed between the PND molecule and two sequence DNA fragments and confirmed the experimental findings. This research sets the stage for developing highly sensitive DNA biosensors that will be used to monitor and quantify toxic herbicides in real samples by fusing the advantages of nanohybrid structures with crucial knowledge from a molecular docking investigation.

Structural variation of GL1 gene determines the trichome formation in Brassica juncea.

KEY MESSAGE: A 448 kb region on chromosome B02 was delimited to be associated with trichome trait in Brassica juncea, in which the BjuVB02G54610 gene with a structural variation of 3 kb structure variation (SV) encoding a MYB transcription factor was predicted as the possible candidate gene. Mustards (Brassica juncea) are allopolyploid crops in the worldwide, and trichomes are essential quality attributes that significantly influence its taste and palpability in vegetable-use cultivars. As important accessory tissues from specialized epidermal cells, trichomes also play an important role in mitigating biotic and abiotic stresses. In this study, we constructed a F2 segregating population using YJ27 with intensive trichome leaves and 03B0307 with glabrous leaves as parents. By bulked segregant analysis (BSA-seq), we obtained a 2.1 Mb candidate region on B02 chromosome associated with the trichome or glabrous trait formation. Then, we used 13 Kompetitive Allele Specific PCR (KASP) markers for fine mapping and finally narrowed down the candidate region to about 448 kb in length. Interestingly, among the region, there was a 3 kb sequence deletion that located on the BjuVB02G54610 gene in the F2 individuals with trichome leaves. Genotyping results of F2 populations confirmed this deletion (R2 = 81.44%) as a major QTL. Natural population re-sequencing analysis and genotyping results further validated the key role of the 3 kb structure variation (SV) of insertion/deletion type in trichome development in B. juncea. Our findings provide important information on the formation of trichomes and potential target gene for breeding vegetable mustards.

A case report of gadopentetate dimeglumine-induced cardiac arrest: Resuscitation using extracorporeal membrane oxygenation.

Gadopentetate dimeglumine (Gd-DTPA) is commonly used for enhancement in magnetic resonance imaging, but rarely causes serious adverse reactions. The patient presented in this report had a cardiac arrest and multiple organ dysfunction syndrome within a short time after administration of Gd-DTPA. Immediately after receiving an intravenous injection of Gd-DTPA, the patient felt nausea and chest tightness, and developed systemic erythema. He was successfully treated using veno-arterial extracorporeal membrane oxygenation (ECMO) combined with continuous renal replacement therapy without any serious complications or neurological deficits. We report a patient who was successfully treated for Gd-DTPA-induced cardiac arrest with ECMO. Thus, ECMO may be an effective treatment for cardiac arrest secondary to anaphylaxis.

A case of multiple thrombosis and septic shock in a critically ill patient with Omicron infection.

The emergence of the acute respiratory syndrome coronavirus 2 variant named Omicron has become a global concern. A 74-year-old unvaccinated patient was critically ill infected with the Omicron variant characterised by septic shock, large-scale cerebral embolism, deep vein thrombosis, and multiple organ dysfunction with respiratory failure, acute renal failure, coagulation dysfunction. The clinical symptoms were successfully controlled by active rescue treatment such as anti-infection, anti-shock, implantation of a vena cava filter as well as multi-organ function support. Although there are many complications in critically ill patients with Omicron variant infections, especially coagulation disorders and thrombosis, they can be resolved with a combination of Chinese and Western medicine positive rescue.

The roles of primary care doctors in the COVID-19 pandemic: consistency and influencing factors of doctor's perception and actions and nominal definitions.

BACKGROUND: At the end of 2019, the Coronavirus Disease 2019 (COVID-19) pandemic broke out. As front-line health professionals, primary care doctors play a significant role in screening SARS-CoV-2 infection and transferring suspected cases. However, the performance of primary care doctors is influenced by their knowledge and role perception. A web-based cross-sectional survey was conducted to assess the consistency and influencing factors of primary care doctor's role perception and expert advice in the guidelines (regulatory definition). METHODS: We designed the questionnaire using "Wenjuanxing" platform, distributed and collected the questionnaire through WeChat social platform, and surveyed 1758 primary care doctors from 11 community health service stations, community health service centers and primary hospitals in Zhejiang Province, China. After the questionnaire was collected, descriptive statistics were made on the characteristics of participants, and univariate analysis and multivariate analysis were used to determine the relevant factors affecting their role cognition. RESULTS: In the reporting and referral suspected cases and patients receiving treatment, most participants' cognition of their roles were consistent with the requirements of guidelines. However, 49.54% and 61.43% of participant doctors were not in line with the government guidelines for diagnosing and classifying COVID-19 and treating suspected cases, respectively. Having a middle or senior professional title and participating in front-line COVID-19 prevention and control work is beneficial to the accurate role perception of diagnosis and classification of COVID-19, the reporting and transfer of suspected cases, and the treatment of suspected cases. CONCLUSIONS: Primary care doctors' role perceptions in the COVID-19 pandemic are not always consistent with government guidelines in some aspects, such as transferring and diagnosing suspected cases. Therefore, it is essential to guide primary care doctors in performing their duties, especially those with lower professional titles.

A natural mutation of the NST1 gene arrests secondary cell wall biosynthesis in the seed coat of a hull-less pumpkin accession.

Hull-less pumpkins (Cucurbita pepo L.) are naturally occurring novel variants known as oilseed or naked-seeded pumpkins, and are characterized by the absence of a normal lignified seed coat. Due to a specialized seed coat structure, these variants serve as a good model for studying seed coat formation and simplify the processing of pumpkin seeds. However, causal genes for this hull-less trait still remain unknown. Here, by bulked segregant analysis and fine mapping, we found that mutation of a single gene, NAC SECONDARY WALL THICKENING PROMOTING FACTOR 1 (NST1), accounts for the hull-less trait. A 14-bp sequence insertion in the CpNST1 gene causes premature termination of CpNST1 translation, leading to lack of secondary cell wall (SCW) biosynthesis in hull-less seed coats. In situ hybridization analysis provided further evidence for the role of CpNST1 in pumpkin seed coat SCW biosynthesis. Interestingly, through secondary cell wall compositional analysis, we found that the main SCW components differed among cell layers in the seed coat. RNA-seq analysis indicated an upstream role of CpNST1 in the SCW biosynthesis network. Collectively, our findings provide mechanistic insight into seed coat SCW biosynthesis, and a target gene for breeders to introduce this hull-less trait for commercial exploitation.

A 215-bp indel at intron I of BoFLC2 affects flowering time in Brassica oleracea var. capitata during vernalization.

KEY MESSAGE: In response to cold, a 215-bp deletion at intron I of BoFLC2 slows its silencing activity by feedback to the core genes of the PHD-PRC2 complex, resulting in late flowering in cabbage. Cabbage is a plant-vernalization-responsive flowering type. In response to cold, BoFLC2 is an important transcription factor, which allows cabbage plants to remain in the vegetative phase. However, there have been few reports on the detailed and functional effects of genetic variation in BoFLC2 on flowering time in cabbage. Herein, BoFLC2E and BoFLC2L, cloned from extremely early and extremely late flowering cabbages, respectively, exhibited a 215-bp indel at intron I, three non-synonymous SNPs and a 3-bp indel at exon II. BoFLC2L was found to be related to late flowering, as verified in 40 extremely early/late flowering accessions, a diverse set of cabbage inbred lines and two F2 generations by using indel-FLC2 marker. Among the genetic variation of BoFLC2, the 215-bp deletion at intron I was the main reason for the delayed flowering time, as verified in the transgenic progenies of seed-vernalization-responsive Arabidopsis thaliana (Col) and rapid cycler B. oleracea (TO1000, boflc2). This is the first report to show that the intron I indel of BoFLC2 affects the flowering time of cabbage. Although the intron I 215-bp indel between BoFLC2E and BoFLC2L did not cause alternative splicing, it slowed BoFLC2L silencing during vernalization and feedback to the core genes of the PHD-PRC2 complex, resulting in their lower transcription levels. Our study not only provides an effective molecular marker-assisted selective strategy for identifying bolting-resistant resources and breeding improved varieties in cabbage, but also provides an entry point for exploring the mechanisms of flowering time in plant-vernalization-responsive plants.

The ratio of superior mesenteric artery diameter to superior mesenteric vein diameter based on non-enhanced computed tomography in the early diagnosis of spontaneous isolated superior mesenteric artery dissection.

BACKGROUND: Spontaneous isolated superior mesenteric artery (SMA) dissection (SISMAD) is a rare cause of abdominal pain. The aim of the study is to investigate the role of a new parameter, the ratio of the SMA diameter to the superior mesenteric vein (SMV) diameter (SMA/SMV) based on non-enhanced computed tomography (CT), in the early diagnosis of SISMAD. METHODS: In a registry study from December 2013 to June 2021, 97 abdominal pain SISMAD patients (SISMAD group) admitted to our hospital were enrolled. Meanwhile, the matched sex and age abdominal pain non-SISMAD patients at 1:2 were collected in reverse chronological order as the control group. Student's t-test, Wilcoxon rank-sum test, and Chi-square test were used to compare differences between the SISMAD and control groups. MedCalc was used to generate receiver operating characteristic (ROC) curve. RESULTS: A total of 291 abdominal pain patients, including 97 SISMAD patients and 194 non-SISMAD patients, were included in the current study. The maximum SMA diameter, perivascular exudation, and SMA/SMV based on non-enhanced CT were significant between the two groups (all P<0.05). ROC curves showed that for the maximum SMA diameter, the area under the curve (AUC), cut-off, sensitivity, and specificity were 0.926, 9.80, 93.8%, and 79.4%, respectively. For SMA/SMV, its AUC, cut-off, sensitivity, and specificity were 0.956, 0.83, 88.7%, and 92.3%, respectively. The diagnostic efficiency of SMA/SMV was better than that of the maximum SMA diameter (P<0.05). The combined parameters of SMA/SMV and maximum SMA diameter had the best diagnostic efficiency (AUC=0.970). CONCLUSION: SMA/SMV may be a potential marker for SISMAD.

Clinical characteristics and misdiagnosis of spontaneous isolated superior mesenteric artery dissection.

BACKGROUND: Spontaneous isolated superior mesenteric artery (SMA) dissection (SISMAD) is a rare disease with a potentially fatal pathology. Due to the lack of specificity of clinical characteristics and laboratory tests, misdiagnosis and missed diagnosis are often reported. Therefore, the aim of this study was to investigate the clinical characteristics and misdiagnosis of SISMAD. METHODS: In a registry study from January 2013 to December 2020, 110 patients with SISMAD admitted to the First Affiliated Hospital of Wenzhou Medical University were enrolled. Descriptive methods were used to analyse clinical characteristics, laboratory data, diagnostic method or proof, misdiagnosed cases, plain computed tomography (CT) findings and dissection features. To study the relationship between dissection features and treatment modality, the selected patients were classified into the conservative group (n = 71) and the non-conservative group (n = 39). The Chi-square test and Student's t-test were used to compare the conservative and non-conservative groups. RESULTS: One hundred ten patients with SISMAD, including 100 (90.9%) males and 10 (9.1%) females, with a mean age of 52.4 ± 7.6 years, were enrolled in the study. Relevant associated comorbidities included a history of hypertension in 43 cases (39.1%), smoking in 46 cases (41.8%), and alcohol consumption in 34 cases (30.9%). One hundred four patients (94.5%) presented with abdominal pain. Abnormalities in the C-reactive protein lever, white blood cells count and D-dimer lever were the 3 most common abnormal findings. There were 32 misdiagnosis or missed diagnosis. Fourteen cases were misdiagnosed because of insufficient awareness. Twelve cases were misdiagnosed because of disease features. Twenty cases were misdiagnosed as SMA embolism. Among them, There were 15 cases of Yun type IIb SISMAD. Sixty-six patients underwent plain CT. The maximum SMA diameter was 12.1 (11.3-13.1) mm, and the maximum SMA diameter was located on the left renal vein (LRV) plane in 68.2% of cases. Dissection features observed on contrast-enhanced CT (CECT), CT angiography (CTA), or digital subtraction angiography (DSA) showed that there were 70 cases (63.6%) of Yun type IIb SISMAD, the maximum SMA diameter was 13.0 ± 2.4 mm, the location of the maximum SMA diameter was on the LRV plane in 64.5% of cases, and 7.3% of cases were complicated with intestinal obstruction, including bowel necrosis in 3.6% of cases. There were differences between the conservative group and non-conservative groups in the residual true lumen diameter or degree of true lumen stenosis and the presence of intestinal obstruction or bowel necrosis (all P < 0.05). CONCLUSION: For SISMAD, misdiagnosis and missed diagnosis were usually caused by insufficient awareness and disease features. SISMAD should be considered in the differential diagnosis of patients presenting with unexplained abdominal pain, especially males, patients in the 5th decade of life, patients with hypertension, and patients with an enlarged SMA diameter or a maximum SMA diameter located on the LRV plane on plain CT. Mesenteric CTA or CECT should be recommended for the investigation of these conditions.

Architecting polyelectrolyte hydrogels with Cu-assisted polydopamine nanoparticles for photothermal antibacterial therapy.

Polydopamine nanoparticles (PDA NPs) are an appealing biomimetic photothermal agent for photothermal antibacterial treatment because of their long-term safety, excellent photostability, accessible manufacturing, and good biodegradability. However, the low photothermal conversion efficiency (PCE) of PDA NPs requires high-power and long-term near-infrared light irradiation, which severely restricts their practical application. In this work, PDA@Cu NPs were fabricated by growing Cu NPs in situ on the surface of PDA and then introduced into a polyelectrolyte hydrogel precursor (cationic polyethyleneimine/anionic pectin, named as CPAP). The formulated photothermal platform possessed a high PCE (55.4%), almost twice as much as pure PDA NPs (30.8%). Moreover, the designed CPAP/PDA@Cu captured and killed some bacteria by electrostatic adsorption, which helped enhance the antibacterial performance. As expected, the formed CPAP/PDA@Cu that combined the advantageous features of PDA@Cu NPs (high PCE) and CPAP matrix (inherent antibacterial activity and preventing NPs aggregation) can efficiently kill bacteria both in vitro and in vivo under the help of near-infrared laser irradiation. Taken together, this study offers a promising strategy for constructing a facile and safe PDA-based photothermal agent for photothermal antibacterial therapy.

Neurological outcomes in adult drowning patients in China.

BACKGROUND: Drowning is the third leading cause of unintentional death worldwide. The epidemiological characteristics of adult drownings are rarely reported. OBJECTIVE: Investigate factors associated with neurological prognosis in adult drowning inpatients. DESIGN: Multicenter medical record review. SETTING: Tertiary health care institutions. PATIENTS AND METHODS: We collected demographic and clinical data on patients who drowned but survived between September 2006 and January 2020. Neurological prognosis was compared in patients with and without cardiac arrest. MAIN OUTCOME MEASURES: Neurological outcomes. SAMPLE SIZE AND CHARACTERISTICS: 142 patients with mean age of 50.6 (19.8) years, male/female ratio of 1.54:1. RESULT: Forty-five patients (31.7%) received CPR, 90 patients (63.4%) experienced unconsciousness, and 59 patients (41.5%) received endotracheal intubation and mechanical ventilation. Multivariate logistic regression analysis showed that the initial blood lactic acid level (OR: 7.67, 95%CI: 1.23-47.82, P=.029) was associated with a poor neurological prognosis in patients without cardiac arrest. The incidence of ICU admission (OR: 16.604, 95%CI: 1.15-239.49, P=.039) was associated with a poor neurologic prognosis in patients with cardiac arrest. CONCLUSIONS: For the drowning patients with cardiac arrest, ICU admission was associated with neurological function prognosis in these patients. Among the patients without cardiac arrest, the initial lactate value was associated with neurological function prognosis of these patients. LIMITATIONS: Retrospective. CONFLICT OF INTEREST: None.

General regulatory factor 3 regulates the expression of alternative oxidase 1a and the biosynthesis of glucosinolates in cytoplasmic male sterile Brassica juncea.

Mitochondrial retrograde signaling (MRS) plays an essential role in sensing and responding to internal and external stimuli to optimize growth to adapt to the prevailing environmental conditions. Previously studies showed alterations on MRS in cytoplasmic male sterile (CMS) plant. However, the regulators involved in MRS in CMS plants remain largely unknown. In this study, we used alternative oxidase 1a (AOX1a) as an indicator of MRS and found that the expression of AOX1a was significantly downregulated in a CMS line comparing to its revertant line, thus indicating an alteration in MRS in the CMS line. By performing a BLAST search of known regulatory components involved in MRS in yeast, we identified general regulatory factor 3 (GRF3), an orthologue of Bmh1/2 in yeast, and demonstrated an association between this gene and MRS in plants, as evidenced by change in AOX1a expression. GRF3 protein was found to be located in the nucleus and the plasma membrane. Further studies showed that GRF3 interacted with MYB29, and regulated the biosynthesis of glucosinolates in Brassica juncea. These findings revealed that GRF3, a negative regulator of AOX1a, is involved in MRS, and also plays a vital role in the accumulation of glucosinolates in CMS crops.

[Mechanism of microRNA-1 regulating H9c2 cardiomyocyte apoptosis after hypoxia/reoxygenation].

OBJECTIVE: To investigate the changes of cardiomyocyte apoptosis after hypoxia/reoxygenation (H/R) regulated by microRNA-1 (miR-1). METHODS: Cardiomyocyte strain H9c2 derived from rat embryonic heart tissue were cultured in vitro. The cells in logarithmic growth phase were divided into blank control group, H/R group, miR-1 mimics+H/R group, miR-1 inhibitor antisense oligonucleotide (ASO)+H/R group and microRNA negative control fragment (miRNA NC)+H/R group. The low sugar DMEM medium containing low concentration of fetal bovine serum (FBS) was used as the medium under anoxic condition. After being cultured in a closed anaerobic incubator at 37 centigrade (95% N2 and 5% CO2) for 12 hours, the cells were cultured with the fresh high sugar DMEM medium containing 5% FBS in a closed incubator at 37 centigrade for reproducing cardiomyocyte H/R model. The blank control group was cultured in high glucose DMEM medium containing 10% FBS in 37 centigrade and 5% CO2 incubator. In miR-1 mimics+H/R group, miR-1 ASO+H/R group and miRNA NC+H/R group, the corresponding transfectants were mixed in high glucose DMEM medium and transfected into cells before H/R model was established, and the final concentration was 50 nmol/L. The blank control group and H/R group were added with DMEM medium at the same time. After the establishment of the model, the expression level of miR-1 was detected by real-time fluorescence quantitative polymerase chain reaction (qPCR). The expression levels of apoptosis-related proteins caspase-9, Bcl-2 and Bax were detected by Western blotting, and cardiomyocyte apoptosis was detected by flow cytometry. RESULTS: Compared with the blank control group, the expression levels of miR-1, caspase-9 and Bax protein and the apoptosis rate of cardiomyocytes were significantly increased, while the expression level of Bcl-2 was significantly decreased, which indicated that the expression of miR-1 and the level of apoptosis were increased in H/R group. Compared with H/R group, the expressions of miR-1, caspase-9 and Bax and the apoptosis rate of cardiomyocytes in miR-1 mimics+H/R group were further increased [miR-1 (2-ΔΔCt): 11.59±1.48 vs. 2.57±0.38, caspase-9 protein (caspase-9/ß-actin): 2.59±0.12 vs. 1.56±0.20, Bax protein (Bax/ß-actin): 4.09±0.38 vs. 1.97±0.13, apoptosis rate: (25.23±0.87)% vs. (17.86±0.73)%, all P < 0.01], while the expression of Bcl-2 was decreased (Bcl-2/ß-actin: 0.37±0.02 vs. 0.49±0.03, P < 0.01). The expressions of miR-1, caspase-9 and Bax and the apoptosis rate were significantly decreased in miR-1 ASO+H/R group [miR-1 (2-ΔΔCt): 1.16±0.06 vs. 2.57±0.38, caspase-9 protein (caspase-9/ß-actin): 1.05±0.24 vs. 1.56±0.20, Bax protein (Bax/ß-actin): 0.93±0.11 vs. 1.97±0.13, apoptosis rate: (11.19±0.85)% vs. (17.86±0.73)%, all P < 0.05], while the expression of Bcl-2 was increased (Bcl-2/ß-actin: 0.84±0.17 vs. 0.49±0.03, P < 0.05). There was no significant difference in miR-1 expression, caspase-9, Bax and Bcl-2 protein expressions, and apoptosis rate between H/R+miRNA NC group and H/R group. CONCLUSIONS: The expression of miR-1 and level of apoptosis were increased in H/R cells, and miR-1 could aggravate cardiomyocyte apoptosis.

Genomic signatures of vegetable and oilseed allopolyploid Brassica juncea and genetic loci controlling the accumulation of glucosinolates.

Allopolyploid Brassica juncea crops in Brassicaceae are becoming increasingly revitalized as vegetables and oilseeds owing to wide adaptability and significant economic values. However, the genomic differentiation of diversified vegetables and oilseed B. juncea and the genetic basis underlying glucosinolates accumulation have yet to be elucidated. To address this knowledge gap, we report the sequencing of pairwise genomes of vegetable and oilseed B. juncea at chromosome scale. Comparative genomics analysis unveils panoramic structural variation footprints, particularly the genetic loci of HSP20 and TGA1 associated with abiotic and biotic stresses responses between oilseed and vegetable subgroups. We anchored two major loci of MYB28 (HAG1) orthologues caused by copy number variations on A02 and A09 chromosomes using scored genomic SNPs-based GWAS that are responsible for seed oil quality-determining glucosinolates biosynthesis. These findings will provide valuable repertories of polyploidy genomic information enabling polyploidy genome evolution studies and precise genomic selections for crucial traits like functional components of glucosinolates in B. juncea crops and beyond.