Machine Learning Prediction Model and Risk Factor Analysis of Reoperation in Recurrent Lumbar Disc Herniation Patients After Percutaneous Endoscopic Lumbar Discectomy.

OBJECTIVE: To investigate the risk factors of reoperation after percutaneous endoscopic lumbar discectomy (PELD) due to recurrent lumbar disc herniation (rLDH) and to establish a set of individualized prediction models. METHODS: Patients who underwent PELD successfully from January 2016 to February 2022 in a single institution were enrolled in this study. Six methods of machine learning (ML) were used to establish an individualized prediction model for reoperation in rLDH patients after PELD, and these models were compared with logistics regression model to select optimal model. RESULTS: A total of 2603 patients were enrolled in this study. 57 patients had repeated operation due to rLDH and 114 patients were selected from the remaining 2546 nonrecurrent patients as matched controls. Multivariate logistic regression analysis showed that disc herniation type (P < .001), Modic changes (type II) (P = .003), sagittal range of motion (sROM) (P = .022), facet orientation (FO) (P = .028) and fat infiltration (FI) (P = .001) were independent risk factors for reoperation in rLDH patients after PELD. The XGBoost AUC was of 90.71%, accuracy was approximately 88.87%, sensitivity was 70.81%, specificity was 97.19%. The traditional logistic regression AUC was 77.4%, accuracy was about 77.73%, sensitivity was 47.15%, specificity was 92.12%. CONCLUSION: This study showed that disc herniation type (extrusion, sequestration), Modic changes (type II), a large sROM, a large FO and high FI were independent risk factors for reoperation in LDH patients after PELD. The prediction efficiency of XGBoost model was higher than traditional Logistic regression analysis model.

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.

Identification of Candidate Genes for Clubroot-Resistance in Brassica oleracea Using Quantitative Trait Loci-Sequencing.

Clubroot caused by Plasmodiophora brassicae is a devastating disease of cabbage (Brassica oleracea). To identify quantitative trait loci (QTLs) for clubroot resistance (CR) in B. oleracea, genomic resequencing was carried out in two sets of extreme pools, group I and group II, which were constructed separately from 110 and 74 F2 cloned lines derived from the cross between clubroot-resistant (R) cabbage "GZ87" (against race 4) and susceptible (S) cabbage "263." Based on the QTL-sequencing (QTL-Seq) analysis of group I and group II, three QTLs (i.e., qCRc7-2, qCRc7-3, and qCRc7-4) were determined on the C07 chromosome. RNA-Seq and qRT-PCR were conducted in the extreme pools of group II before and after inoculation, and two potential candidate genes (i.e., Bol037115 and Bol042270), which exhibiting upregulation after inoculation in the R pool but downregulation in the S pool, were identified from the three QTLs on C07. A functional marker "SWU-OA" was developed from qCRc7-4 on C07, exhibiting ∼95% accuracy in identifying CR in 56 F2 lines. Our study will provide valuable information on resistance genes against P. brassicae and may accelerate the breeding process of B. oleracea with CR.

Quantitative reconstruction of the complex-valued object based on complementary phase modulations.

In the coherent diffraction imaging (CDI) techniques, a key point is to reconstruct the complex-valued object from the far-field intensity measurements, i.e., solving the phase retrieval problem. However, due to this ill-posed problem, traditional phase retrieval algorithms often encounter some problems associated with the iteration convergence. In this work, complementary phase modulations (CPM) are introduced to generate different far-field intensity measurements. The namely CPM-based method aims to find out the global optimal solution by imposing multi-dimensional constraints, including the diverse intensity images at the Fourier plane and the CPM at the object plane. It is proved by the numerical simulations and the optical experiments that the convergence speed and the recovery accuracy could be greatly improved. Furthermore, the shifting complementary phase modulations (SCPM)-based method is proposed by introducing more CPMs. The reconstruction performance is further improved even when the phase range is larger, and the support constraints are not required. In addition, the SCPM-based method is more robust to the Poisson noise. With the outstanding reconstruction performance, the CPM-based methods may be helpful to phase imaging in the application of visible-light microscopy and X-ray imaging.

Performance of single-stage partial nitritation and anammox reactor treating low-phenol/ammonia ratio wastewater and analysis of microbial community structure.

Phenol and ammonia are common pollutants in many industrial wastewaters. The partial nitritation and anammox process is a very promising technology for treating phenol-ammonia wastewater. This study was the first time to rapidly achieve the start-up and operation of the single-stage partial nitritation /anammox reactor treating phenol-ammonia wastewater. The optimized ratio of phenol and nitrogen (phenol/NH4 + -N=0.3) was set to start-up the reactor. After 60 days of operation, the total nitrogen and COD removal efficiencies were around 73.0% and 79.5%, respectively. The activity of ammonium-oxidizing bacteria was291.1 ± 3.0 mg NH4 + -N g-1 MLVSS d-1 and the specific anammox activity was 20.9 ± 1.0 mg NH4 + -N g-1 MLVSS d-1 . The results indicated that the anammox bacteria had adapted to phenol condition and remained stable activity after the 60 days' operation in the reactor. The sequence analysis of 16SrRNA showed that the microbial community structure evolved to a balanced distribution that the removals of phenol and ammonia could be achieved simultaneously. PRACTITIONER POINTS: Phenol/N ratio of 0.3 was set to start up the single-stage partial nitritation/anammox reactor. The single-stage partial nitritation /anammox reactor was rapidly started up when treating the phenol-ammonia wastewater. Total nitrogen removal rate and COD removal efficiencies could achieve to 73.0% and 79.5%, respectively. Microbial community structure evolved to stable distribution of which AOB, anammox bacteria, denitrification bacteria and heterotrophic nitrification bacteria coexisted.

Development of Ogura CMS restorers in Brassica oleracea subspecies via direct RfoB gene transformation.

KEY MESSAGE: The Ogura CMS RfoB restorer developing via RfoB gene transformation was utilized to produce specific morphological Ogura CMS restorers and clubroot resistance lines in Brassica oleracea subspecies. Brassica oleracea vegetables including cabbage, cauliflower, kohlrabi, Brussels sprouts and Chinese kale are morphologically very different despite being members of the same species. The Ogura cytoplasmic male sterility (CMS) system is the most stable strategy for the hybrid breeding of these species. However, this limits the utilization of some excellent genes due to the lack of fertile restorer genes in the system. Herein, to efficaciously use Ogura CMS, the Ogura CMS RfoB restorer was produced by transforming the modified RfoB restorer gene into the Ogura CMS line 'CMS2016' of B. oleracea var. capitata. This gene was shown to recover fertility of natural Ogura CMS lines in B. oleracea subspecies and create transient Ogura CMS RfoB restorers such as the clubroot resistance Ogura CMS RfoB restorer. Interestingly, clubroot resistant individuals without transgenic elements were screened in the progenies of hybrids between B. oleracea inbred lines and the clubroot resistance Ogura CMS RfoB restorer. In addition, 18 different morphological Ogura CMS restorers were developed to specifically recover fertile of Ogura CMS cultivars in B. oleracea subspecies.

Influence of immersion depth of membrane on filtration performance of anaerobic membrane bioreactor.

Membrane fouling is still the main obstacle that hinders the development and implementation of anaerobic membrane bioreactor (AnMBR). In conventional upflow anaerobic reactors, sludge at different height usually presents certain differences in characteristics in terms of particle size, etc. The immersion depth of membrane modules in anaerobic reactors can also influence the fouling of membrane. Thus, it is of great interest to investigate the fouling mechanism with the membrane installed at different heights in reactors. The filtration performance and sludge properties were investigated at different heights of AnMBR. The fouling of membrane in the middle position was severer than that in the top and bottom positions. The total resistance of membrane in the top, middle, and bottom positions was 27.31 × 1011 m-1, 34.67 × 1011 m-1, and 25.29 × 1011 m-1, respectively. By comparing the characteristics and structure of bulk sludge and cake layer at three heights, the bulk sludge in the middle position presented higher content of soluble microbial products (SMP) and finer flocs, and the cake layer was also denser. The results obtained in this study indicated that small size of sludge flocs as well as adhesion of SMP might be the major factors governing membrane fouling at different height in the AnMBR.

Identification and characterization of BoPUB3: a novel interaction protein with S-locus receptor kinase in Brassica oleracea L.

Armadillo repeat containing 1 (ARC1) is phosphorylated by S-locus receptor kinase (SRK) and functions as a positive regulator in self-incompatibility response of Brassica. However, ARC1 only causes partial breakdown of the self-incompatibility response, and other SRK downstream factors may also participate in the self-incompatibility signaling pathway. In the present study, to search for SRK downstream targets, a plant U-box protein 3 (BoPUB3) was identified from the stigma of Brassica oleracea L. BoPUB3 was highly expressed in the stigma, and its expression was increased with the stigma development and reached to the highest level in the mature-stage stigma. BoPUB3, a 76.8-kDa protein with 697 amino acids, is a member of the PUB-ARM family and contains three domain characteristics of BoARC1, including a U-box N-terminal domain, a U-box motif, and a C-terminal arm repeat domain. The phylogenic tree showed that BoPUB3 was close to BoARC1. The synteny analysis revealed that B. oleracea chromosomal region containing BoPUB3 had high synteny with the Arabidopsis thaliana chromosomal region containing AtPUB3 (At3G54790). In addition, the subcellular localization analysis showed that BoPUB3 primarily localized in the plasma membrane and also in the cytoplasm. The combination of the yeast two-hybrid and in vitro binding assay showed that both BoPUB3 and BoARC1 could interact with SRK kinase domain, and SRK showed much higher level of ß-galactosidase activity in its interaction with BoPUB3 than with BoARC1. These results implied that BoPUB3 is a novel interactor with SRK, which lays a basis for further research on whether PUB3 participates in the self-incompatibility signaling pathway.

Efficacy and mechanism of acupuncture for ischemic poststroke depression: Study protocol for a multicenter single-blinded randomized sham-controlled trial.

INTRODUCTION: Poststroke depression is a serious and common complication of stroke, especially the ischemic poststroke depression. Antidepressants are used in poststroke depression, and acupuncture may be an alternative approach. However, the efficacy and mechanism of acupuncture for poststroke depression has not been confirmed. METHODS/DESIGN: This is a multicenter, central-randomized, single-blind, sham-controlled clinical trial. We will allocate 208 subjects aged between 40 and 80 years old, diagnosed with initial poststroke depression (PSD) within 6 months to 2 groups randomly in a ratio of 1:1. Patients in the experimental group will be treated with traditional acupuncture and placebo pills, whereas the others in the control group will be treated with sham-acupoints acupuncture and antidepressant (fluoxetine hydrochloride tablets). All will be given acupuncture and/or medication treatment for 12 weeks, and then received 12-week follow-up. Patients will be evaluated with the 17-item Hamilton Depression Scale and Se1f-rating Depression Scale for depression state, National Institute of Health Stroke Scale for neurological deficit, Modified Barthel Index for activities of daily living, Treatment Emergent Symptom Scale for side effects of treatments, diagnosis and evaluation criteria of traditional Chinese medicine for stroke (try out) for curative effects of stroke, and clinical global impression for synthesize effect before and the 2nd, 4th, 8th, and 12th week of treatment, 24th week of follow-up. Study on mechanisms of acupuncture will be revealed through the diversity of brain metabolites (choline-containing compounds [Cho], N-acetylaspartate [NAA], myoinositol, glutamine and glutamate complex, creatine [Cr], Cho/Cr, Cho/NAA, Cr/NAA) in bilateral dorsolateral prefrontal cortex and anterior cingulate cortex monitored by proton magnetic resonance spectroscopy, and serum monoamine neurotransmitters (5-hydroxytryptamine, norepinephrine, dopamine) and cytokines (brain-derived neurotrophic factor [BDNF], interleukin [IL]-4, IL-6, IL-10, IL-18, IL-1ß, tumor necrosis factor alpha) before and the 12th week of treatment. Baseline characteristics of patients will be summarized by groups and compared with chi-square for categorical variables, and 2-sample t tests or Wilcoxon rank-sum test for the continuous variables. Primary and secondary outcomes according to the measurement times are applicable to univariate repetitive measurement deviation analysis or 2-sample t tests, or Wilcoxon rank-sum test. CONCLUSION: The present research is designed to investigate efficacy and mechanism of traditional acupuncture therapy on ischemic PSD, also to explore the correlation between cerebra metabolic and serologic factors, and ischemic PSD. With this research, we are looking forward to find out an appropriate alternative nondrug therapy for PSD people to alleviate the adverse effects and drug dependence caused by antidepressants.

CRISPR/Cas9-mediated multiple gene editing in Brassica oleracea var. capitata using the endogenous tRNA-processing system.

Cabbage (Brassica oleracea var. capitata) is a biennial plant with strong self-incompatibility and an obligate requirement for prolonged vernalization by exposure to low temperatures to induce flowering. These characteristics significantly increase the difficulty of exploiting novel germplasm induced by physical or chemical mutagens. In this study, we report a CRISPR/Cas9 gene-editing system based on endogenous tRNA processing to induce high efficiency and inheritable mutagenesis in cabbage. Using the phytoene desaturase gene BoPDS, the S-receptor kinase gene BoSRK, and the male-sterility-associated gene BoMS1 as the target genes, multisite and multiple gene mutations were achieved using a construct with tandemly arrayed tRNA-sgRNA architecture to express multiple sgRNAs. The BoSRK3 gene mutation suppressed self-incompatibility completely, converting the self-incompatible line into a self-compatible line. In addition, the BoMS1 gene mutation produced a completely male-sterile mutant, which was highly cross compatible with its nonmutant isoline at the flowering stage as a result of a simultaneous BoSRK3 gene mutation, enabling the economic propagation of the male-sterile line through bee-mediated cross-pollination. Interestingly, higher site mutation efficiency was detected when a guide sequence was inserted into a location in the tandemly arrayed tRNA-sgRNA architecture that was distal from the upstream Pol III promoter. In addition, mutation sites were also detected in the paralogous genes of the BoPDS and BoSRK genes that had fully consistent sequences or base mismatches but beyond the "seed" region in the spacer sequence compared with the target sgRNAs. Collectively, our results demonstrate that the CRISPR/Cas9 system, coupled with an endogenous tRNA-processing system, is an efficient tool to improve cabbage traits.

Performance and recovery of a completely separated partial nitritation and anammox process treating phenol-containing wastewater.

Anammox process is considered as a promising technology for removing total nitrogen from low-strength ammonium and phenol-containing wastewater. However, it is still a challenge for the anammox process to treat high-strength ammonium and phenol-containing wastewater. A completely separated partial nitritation and anammox (CSPN/A) process was developed to remove total nitrogen from high-strength phenol-containing wastewater. About 92% of COD, 100% of phenol, and 82.4% of total nitrogen were successfully removed at a NH4+-N concentration of 200 mg L-1 with a phenol/NH4+-N mass ratio of 0.5 in the CSPN/A process. Furthermore, a shock loading of 300 mg phenol L-1 with a phenol/NH4+-N mass ratio of 1.5 led to a complete failure of partial nitritation, but the performance was rapidly recovered by the increase of NH4+-N concentration. Although the activities of ammonium-oxidizing bacteria and anammox bacteria were severely inhibited at a phenol/NH4+-N mass ratio of 1.5, the enrichment of efficient phenol degraders in the CSPN stage could strengthen the performance robustness of partial nitritation and anammox process. Therefore, this study presented a new insight on the feasibility of the anammox process for treating high-strength ammonium and phenol-containing wastewater.

Identification of Quantitative Trait Loci for Clubroot Resistance in Brassica oleracea With the Use of Brassica SNP Microarray.

Increasing clubroot resistance (CR) of Brassica oleracea by ascertaining the molecular mechanisms has been the key focus in modern B. oleracea breeding. In order to identify the quantitative trait loci (QTLs) associated with CR in B. oleracea, 94 F2 vegetative lines which were developed by tissue culture of selfed seeds from the F1 generation between a clubroot-resistant B. oleracea inbred line and a susceptible line, were identified for disease incidence and six CR-associated traits under a lab inoculation by Plasmodiophora brassicae and were genotyped with the 60K Brassica SNP array. Significant correlations were detected for numbers of fibrous roots and P. brassicae content in roots with disease incidence. Nine linkage groups were constructed from 565 bins which covered around 3,000 SNPs, spanning 1,028 cM of the B. oleracea genome with an average distance of 1.82 cM between adjacent bins. A total of 23 QTLs were identified for disease incidence and the other two correlated traits, individually explaining 6.1-17.8% of the phenotypic variation. Several overlaps were detected among traits, including one three-traits-overlapped locus on linkage group C08 and two important overlapped regions between the two CR-associated traits on C06. The QTLs were compared with known CR loci/genes and the novelty of our QTLs was discussed.

Rapid establishment of phenol- and quinoline-degrading consortia driven by the scoured cake layer in an anaerobic baffled ceramic membrane bioreactor.

Although toxic and refractory organics, such as phenol and quinoline, are decomposed by anaerobic bacteria, the establishment of specific degrading consortia is a relatively slow process. An anaerobic membrane bioreactor allows for complete biomass retention that can aid the establishment of phenol- and quinoline-degrading consortia. In this study, the anaerobic digestion of phenol (500 mg L-1) and quinoline (50 mg L-1) was investigated using an anaerobic baffled ceramic membrane bioreactor (ABCMBR). The results showed that, within 30 days, 99% of phenol, 98% of quinoline and 88% of chemical oxygen demand (COD) were removed. The substrate utilisation rates of the cake layer for phenol and quinoline, and specific methanogenic activity of the cake layer, were 7.58 mg phenol g-1 mixed liquor volatile suspended solids (MLVSS) day-1, 8.23 mg quinoline g-1 MLVSS day-1 and 0.55 g CODCH4 g-1 MLVSS day-1, respectively. The contribution of the cake layer to the removals of phenol and quinoline was extremely underestimated because the uncounted scoured cake layer was disregarded. Syntrophus was the key population for phenol and quinoline degradation, and it was more abundant in the cake layer than in the bulk sludge. The highly active scattered cake layer sped up the establishment of phenol- and quinoline-degrading consortia in the ABCMBR.

Inhibition of ammonia on anaerobic digestion of synthetic coal gasification wastewater and recovery using struvite precipitation.

Coal gasification wastewater (CGW) contains very high concentrations of phenols and ammonia. However, the potential impact of ammonia on the anaerobic digestion of phenols remained unclear. Firstly, the methanogens and phenols degraders had a good tolerance up to 1gL-1 of total ammonia nitrogen (TAN), but the substrate utilization rate for phenol, and specific methanogenic activity of sludge were decreased by 89% and 67% at 5g TAN L-1, and 94% and 100% at 10g TAN L-1, respectively. Secondly, the optimum struvite crystallization conditions (pH=8.5, 10g TAN L-1, n(Mg2+):n(TAN):n(PO43-)=1:1:1) were obtained by the orthogonal tests. Thirdly, the removal efficiencies of chemical oxygen demand (COD) and phenols were recovered to around 82% and 66%, respectively in the upflow anaerobic sludge blanket reactor using the pretreatment of struvite precipitation at 10g TAN L-1 and 1g phenols L-1. Therefore, anaerobic digestion coupled with struvite precipitation was considered as an alternative way for CGW treatment.

Dissecting Pistil Responses to Incompatible and Compatible Pollen in Self-Incompatibility Brassica oleracea Using Comparative Proteomics.

Angiosperms have developed self-incompatibility (SI) systems to reject self-pollen, thereby promoting outcrossing. The Brassicaceae belongs to typical sporophytic system, having a single S-locus controlled SI response, and was chosen as a model system to study SI-related intercellular signal transduction. In this regard, the downstream factor of EXO70A1 was unknown. Here, protein two-dimensional electrophoresis (2-DE) method and coupled with matrix-assisted laser desorption ionization/time of flight of flight mass spectrometry (MALDI-TOF -MS) and peptide mass fingerprinting (PMF) was used to further explore the mechanism of SI responses in Brassica oleracea L. var. capitata L. at protein level. To further confirm the time point of protein profile change, total proteins were collected from B. oleracea pistils at 0 min, 1 h, and 2 h after self-pollination. In total 902, 1088 and 1023 protein spots were separated in 0 min, 1 h and 2 h 2-DE maps, respectively. Our analyses of self-pollination profiles indicated that proteins mainly changed at 1 h post-pollination in B. oleracea. Moreover, 1077 protein spots were separated in cross-pollinated 1 h (CP) pistil 2-DE map. MALDI-TOF-MS and PMF successfully identified 34 differentially-expressed proteins (DEPs) in SP and CP 1 h 2-DE maps. Gene ontology and KEGG analysis revealed an array of proteins grouped in the following categories: stress and defense response (35%), protein metabolism (18%), carbohydrate and energy metabolism (12%), regulation of translation (9%), pollen tube development (12%), transport (9%) and cytoskeletal (6%). Sets of DEPs identified specifically in SP or only up-regulated expressed in CP pistils were chosen for funther investigating in floral organs and during the process of self- and cross-pollination. The function of these DEPs in terms of their potential involvement in SI in B. oleracea is discussed.

Tongguan Liqiao acupuncture therapy improves dysphagia after brainstem stroke.

Tongguan Liqiao acupuncture therapy has been shown to effectively treat dysphagia after stroke-based pseudobulbar paralysis. We presumed that this therapy would be effective for dysphagia after bulbar paralysis in patients with brainstem infarction. Sixty-four patients with dysphagia following brainstem infarction were recruited and divided into a medulla oblongata infarction group (n = 22), a midbrain and pons infarction group (n = 16), and a multiple cerebral infarction group (n = 26) according to their magnetic resonance imaging results. All patients received Tongguan Liqiao acupuncture for 28 days. The main acupoints were Neiguan (PC6), Renzhong (DU26), Sanyinjiao (SP6), Fengchi (GB20), Wangu (GB12), and Yifeng (SJ17). Furthermore, the posterior pharyngeal wall was pricked. Before and after treatment, patient swallowing functions were evaluated with the Kubota Water Test, Fujishima Ichiro Rating Scale, and the Standard Swallowing Assessment. The Barthel Index was also used to evaluate their quality of life. Results showed that after 28 days of treatment, scores on the Kubota Water Test and Standard Swallowing Assessment had decreased, but scores on the Fujishima Ichiro Rating Scale and Barthel Index had increased in each group. The total efficacy rate was 92.2% after treatment, and was most obvious in patients with medulla oblongata infarction (95.9%). These findings suggest that Tongguan Liqiao acupuncture therapy can repair the connection of upper motor neurons to the medulla oblongata motor nucleus, promote the recovery of brainstem infarction, and improve patient's swallowing ability and quality of life.