New strategy for intraoperative phonosurgical management of recurrent laryngeal nerve infiltrated by thyroid carcinoma.

PURPOSE: Treating an infiltration of the recurrent laryngeal nerve (RLN) by thyroid carcinoma remains a subject of ongoing debate. Therefore, this study aims to provide a novel strategy for intraoperative phenosurgical management of RLN infiltrated by thyroid carcinoma. METHODS: Forty-two patients with thyroid carcinoma infiltrating the RLN were recruited for this study and divided into three groups. Group A comprised six individuals with medullary thyroid cancer who underwent RLN resection and arytenoid adduction. Group B consisted of 29 differentiated thyroid cancer (DTC)patients who underwent RLN resection and ansa cervicalis (ACN)-to-RLN anastomosis. Group C included seven patients whose RLN was preserved. RESULTS: The videostroboscopic analysis and voice assessment collectively indicated substantial improvements in voice quality for patients in Groups A and B one year post-surgery. Additionally, the shaving technique maintained a normal or near-normal voice in Group C one year post-surgery. CONCLUSION: The new intraoperative phonosurgical strategy is as follows: Resection of the affected RLN and arytenoid adduction is required in cases of medullary or anaplastic carcinoma, regardless of preoperative RLN function. Suppose RLN is found infiltrated by well-differentiated thyroid cancer (WDTC) during surgery, and the RLN is preoperatively paralyzed, we recommend performing resection the involved RLN and ACN-to-RLN anastomosis immediately during surgery. If vocal folds exhibit normal mobility preoperatively, the MACIS scoring system is used to assess patient risk stratification. When the MACIS score > 6.99, resection of the involved RLN and immediate ACN-to-RLN anastomosis were performed. RLN preservation was limited to patients with MACIS scores ≤ 6.99.

Co-production of surfactin and fengycin by Bacillus subtilis BBW1542 isolated from marine sediment: a promising biocontrol agent against foodborne pathogens.

Pathogenic bacteria contaminations and related diseases in food industries is an urgent issue to solve. The present study aimed to explore natural food biopreservatives from microorganisms. Using dilution-plate method, a strain BBW1542 with antimicrobial activities against various foodborne pathogenic bacteria was isolated from the seabed silt of Beibu Gulf, which was identified as Bacillus subtilis by the morphological observation and 16S rDNA sequences. The antimicrobial substances of B. subtilis BBW1542 exhibited an excellent stability under cool/heat treatment, UV irradiation, acid/alkali treatment, and protease hydrolysis. The genome sequencing analysis and antiSMASH prediction indicated that B. subtilis BBW1542 contained the gene cluster encoding lipopeptides and bacteriocin subtilosin A. MALDI-TOF-MS analysis showed that the lipopeptides from B. subtilis BBW1542 contained C14 and C15 surfactin homologues, together with fengycin homologues of C18 fengycin A/C16 fengycin B and C19 fengycin A/C17 fengycin B. In silico analysis showed that an eight-gene (sboA-albABCDEFG) operon was involved in the biosynthesis of subtilosin A in B. subtilis BBW1542, and the encoded subtilosin A presented an evident closed-loop structure containing 35 amino acids with a molecular weight of 3425.94 Da. Overall, the antagonistic B. subtilis BBW1542 displayed significant resource value and offered a promising alternative in development of food biopreservation. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05864-3.

Oral opening training increases oral opening in patients with oral submucous fibrosis. / å¼ å£è®­ç»ƒå¢žåŠ å£è ”é»è†œä¸‹çº¤ç»´æ€§å˜æ‚£è€ çš„å¼ å£åº¦.

OBJECTIVES: The mouth restriction of patients with oral submucous fibrosis (OSF) seriously affects their eating food and the quality of life. There are few reports about improving the oral opening degree in patients with OSF. This study aims to explore the effect of oral opening training on the improvement of mouth opening limitation in patients with OSF treated with local injection. METHODS: A total of 220 outpatients with limited mouth opening of OSF were collected from the Center of Stomatology, Xiangya Hospital, Central South University, and randomly divided into a control group and an experiment group (n=110). The control group were treated with local injection of Salvia miltiorrhiza and triamcinolone acetonide, once a week, and 8 times a course. The experimental group were treated with local injection combined with mouth opening training for 2 years. The degree of mouth opening was compared between the 2 groups at the end of local injection treatment, 1 year and 2 years after the treatment. The curative effect was evaluated according to the size of the opening, the lamellar structure of the mucosa, and the condition of the cords. RESULTS: A total of 197 patients completed the whole course of treatment, with 107 in the experimental group and 90 in the control group. At the end of treatment, 1 year and 2 years after the treatment, the degree of mouth opening in the experimental group was (36.14±2.62), (39.67±2.67), and (39.80±2.57) mm, respectively, which was significantly higher than that in the control group (24.71±1.97), (22.82±2.13), and (22.02±2.09) mm, respectively. The difference was significant (P<0.05). The increase of mouth opening in the experimental group was significantly better than that in the control group. Two years after local injection treatment, the effective rate of the experimental group was 97.1%, which was significantly higher than that of the control group (47.8%, P<0.05). CONCLUSIONS: Mouth opening training can significantly increase the degree of mouth opening in patients with OSF treated with local injection.

[Clinical analysis for oral mucosal disease in 21 972 cases].

OBJECTIVE: To analyze the incidence and distribution of oral mucosal diseases in Hunan Province and provide reference for prevention and treatment.
 Methods: The clinical data for all patients, who were treated in Xiangya Hospital of Central South University from April 2013 to March 2017, were collected. After screening, weighing and classifying, sex and age distribution for the disease was analyzed.
 Results: The female with the age between 40 to 49 were in the majority among 21 972 patients. The ratio between men to women was 1:1.05. According to the classification of diseases, the most common diseases were as follows: recurrent aphthous ulcer (27.17%), burning mouth syndrome (15.72%), oral submucous fibrosis (14.75%), oral lichen planus (10.38%), oral leukoplakia (4.21%), traumatic ulceration (4.14%), chronic cheilitis (3.47%), oral fungal infection (3.26%), and atrophic glossitis (2.74%). Recurrent oral ulcer (28.65%), burning mouth syndrome (23.70%) and oral lichen planus (13.31%) were the most common 3 kinds of oral mucosal diseases during females in Hunan. Oral submucous fibrosis was the most common oral mucosal disease among males in Hunan (28.56%).
 Conclusion: Recurrent oral ulcer, burning mouth syndrome and oral lichen planus are very popular in women in Hunan Province, and oral submucous fibrosis is the most common disease in male in this region. It shows a high trend of incidence in the surrounding provinces.

Selective TBK1/IKKi dual inhibitors with anticancer potency.

Increasing evidence suggests that the noncanonical IKKs play critical roles in tumor genesis and development, leading to the notion that noncanonical IKKs may be good targets for cancer therapy. Here, we demonstrate that although TBK1 is not overexpressed or constitutively activated in some tumor cells, targeting IKKi induces the activation of TBK1. Therefore, simultaneously targeting both kinases is necessary to efficiently suppress tumor cell proliferation. We show that three TBK1/IKKi dual inhibitors, which are based on a structurally rigid 2-amino-4-(3'-cyano-4'-pyrrolidine)phenyl-pyrimidine scaffold, potently inhibit cell viability in human breast, prostate and oral cancer cell lines. Treatment with these TBK1/IKKi dual inhibitors significantly impairs tumor development in xenograft and allograft mouse models. The anticancer function of these inhibitors may be partially due to their suppression of TBK1/IKKi-mediated AKT phosphorylation and VEGF expression. Most importantly, these TBK1/IKKi dual inhibitors have drug-like properties including low molecular weight, low cytochrome P450 inhibition and high metabolic stability. Therefore, our studies provide proof of concept for further drug discovery efforts that may lead to novel strategies and new therapeutics for the treatment of human cancer.

Morphological change and genome-wide transcript analysis of Haloxylon ammodendron leaf development reveals morphological characteristics and genes associated with the different C3 and C4 photosynthetic metabolic pathways.

C4 photosynthesis outperforms C3 photosynthesis in natural ecosystems by maintaining a high photosynthetic rate and affording higher water-use and nitrogen-use efficiencies. C4 plants can survive in environments with poor living conditions, such as high temperatures and arid regions, and will be crucial to ecological and agricultural security in the face of global climate change in the future. However, the genetic architecture of C4 photosynthesis remains largely unclear, especially the genetic regulation of C4 Kranz anatomy. Haloxylon ammodendron is an important afforestation tree species and a valuable C4 wood plant in the desert region. The unique characteristic of H. ammodendron is that, during the seedling stage, it utilizes C3 photosynthesis, while in mature assimilating shoots (maAS), it switches to the C4 pathway. This makes an exceptional opportunity for studying the development of the C4 Kranz anatomy and metabolic pathways within individual plants (identical genome). To provide broader insight into the regulation of Kranz anatomy and non-Kranz leaves of the C4 plant H. ammodendron, carbon isotope values, anatomical sections and transcriptome analyses were used to better understand the molecular and cellular processes related to the development of C4 Kranz anatomy. This study revealed that H. ammodendron conducts C3 in the cotyledon before it switches to C4 in AS. However, the switching requires a developmental process. Stable carbon isotope discrimination measurements on three different developmental stages showed that young AS have a C3-like δ13C even though C4 Kranz anatomy is found, which is inconsistent with the anatomical findings. A C4-like δ13C can be measured in AS until they are mature. The expression analysis of C4 key genes also showed that the maAS exhibited higher expression than the young AS. In addition, many genes that may be related to the development of Kranz anatomy were screened. Comparison of gene expression patterns with respect to anatomy during leaf ontogeny provided insight into the genetic features of Kranz anatomy. This study helps with our understanding of the development of Kranz anatomy and provides future directions for studies on key C4 regulatory genes.

An efficient preparation process of sisal fibers via the specialized retting microorganisms: Based on the ideal combination of degumming-related enzymes for the effective removal of non-cellulosic macromolecules.

Bioaugmentation retting with the specialized pectinolytic and xylanolytic microorganisms can accelerate the removal of non-cellulosic macromolecules around plant fibers, thus shortening retting time and facilitating fiber quality. Currently, few specialized microorganisms have been explored for the retting of sisal fibers. The present study excavated the retting fungi including Aspergillus micronesiensis HD 3-6, Penicillium citrinum HD 3-12-3, and Cladosporium sp. HD 4-13 from the region-specific soil samples of planting sisal, and investigated their bioaugmentation retting effects on raw sisal leaves. Results showed that combination of the three fungi achieved the most excellent degumming efficiency (13.69 % of residual gum in sisal fibers) and the highest fiber yield (4.47 %). Furthermore, this fungi combination had the ideal enzymatic hydrolysis features with high activities of pectinase, xylanase and mannanase whereas a low activity of cellulase during the whole retting process, thus endowing the prepared sisal fibers with the lowest mass percentage of non-cellulosic macromolecules (9.76 wt%) and the highest cellulose content (89.23 wt%). SEM and FT-IR analysis further verified that the non-cellulosic substances around sisal fibers were efficiently removed. In summary, the consortia of the three fungi achieved ideal degumming-related enzymes for the removal of non-cellulosic macromolecules, thus acquiring the efficient preparation of sisal fibers.

Chitin Deacetylase from Bacillus aryabhattai TCI-16: Heterologous Expression, Characterization, and Deacetylation Performance.

Chitin deacetylase (CDA) removes the acetyl group from the chitin molecule to generate chitosan in a uniform, high-quality deacetylation pattern. Herein, BaCDA was a novel CDA discovered from our previously isolated Bacillus aryabhattai strain TCI-16, which was excavated from mangrove soil. The gene BaCDA was cloned and overexpressed in Escherichia coli BL21 (DE3) to facilitate its subsequent purification. The purified recombinant protein BaCDA was obtained at a concentration of about 1.2 mg/mL after Ni2+ affinity chromatography. The molecular weight of BaCDA was around 28 kDa according to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. In addition, BaCDA exhibited a significant deacetylation effect on colloidal chitin, and the deacetylation degree was measured from the initial 25.69 to 69.23% by Fourier transform infrared (FT-IR) spectroscopy. Scanning electron microscopy (SEM) observation showed that the surface of colloidal chitin after enzymatic digestion was rough, the crystal fibers disappeared, and the chitin structure was loose and porous with grooves. The results of electrospray ionization mass spectrometry (ESI-MS) showed that BaCDA had full-deacetylation activity against (GlcNAc)4. Molecular docking revealed that BaCDA had an open active pocket capable of binding to the GlcNAc unit. This study not only provides a novel enzymatic resource for the green and efficient application of chitin but also helps to deepen the understanding of the catalytic mechanism of CDA.

[Evaluation of the effect of endoscopic CO2laser arytenoidectomy on re-operation for failed posterior cordotomy].

Objective:To analyze the efficacy of endoscopic CO2 laser arytenoidectomy in treating bilateral vocal cord paralysis. Methods:Ninety-five patients who underwent endoscopic CO2 laser arytenoidectomy for bilateral vocal cord paralysis at the First Affiliated Hospital of Naval Medical University（Changhai Hospital） of Shanghai from January 2009 to December 2022 were included in this study. Among them, 59 patients underwent endoscopic CO2 laser arytenoidectomy as their first glottic enlargement surgery（Group A）, while 36 patients, who had previously undergone two unsuccessful CO2 laser-assisted posterior cordotomies, underwent endoscopic CO2 laser arytenoidectomy as a subsequent surgery（Group B）. Swallowing function, electronic laryngoscopy, perceptual voice evaluation, and objective voice analysis indicators were statistically analyzed before and after surgery to evaluate clinical efficacy. Results:The extubation rate after the first surgery was 84.75% in Group A and 86.11% in Group B, with total extubation rates of 94.92% and 94.44%, respectively. There were no significant differences between the two groups in preoperative and postoperative swallowing function, glottic size, or various voice evaluation indicators（P>0.05）. Within-group comparisons showed that postoperative swallowing function, glottic closure during phonation, perceptual evaluations of G（grade of hoarseness）, A（asthenia）, and B（breathiness） significantly worsened, with increased grades. The maximum transverse diameter of the posterior glottis during inspiration significantly increased, and the VHI-10 score was significantly higher postoperatively. Jitter, shimmer, and the harmonics-to-noise ratio significantly deteriorated, and maximum phonation time significantly shortened（P<0.05）. No significant differences were observed in postoperative R（roughness） and S（strain） compared to preoperative values（P>0.05）. Conclusion:Endoscopic CO2 laser arytenoidectomy can impair voice quality to some extent but effectively alleviates breathing difficulties in patients with bilateral vocal cord paralysis. For patients who did not achieve successful extubation with CO2 laser-assisted posterior cordotomy, endoscopic CO2laser arytenoidectomy is an effective reoperative method, ensuring a high extubation rate while preserving certain voice functions.

[Efficacy analysis of ansa cervicalis anterior root-recurrent laryngeal nerve anastomosis in the treatment of unilateral vocal cord paralysis].

Objective:Investigates the application and clinical efficacy of ansa cervicalis anterior root-recurrent laryngeal nerve（RLN） anastomosis in the treatment of unilateral vocal fold paralysis（UVFP）. Methods:A prospective study was conducted with 92 UVFP patients admitted to our department from January 2018 to January 2022 who received ansa cervicalis anterior root-RLN anastomosis. The course of nerve injury ranged from 6 to 24 months. Videostroboscopy, voice subjective auditory perceptual assessment（GRBAS）, Voice Handicap Index（VHI-10）, voice objective acoustic analysis and laryngeal electromyography（EMG） were used to evaluate the efficacy of the operation. Results:Videostroboscopy showed that although the movement of vocal cords did not return to normal 12 months after operation, their volume and muscle tension were significantly improved and their positions were adducted to the median or near-median. Also the glottic closure, vocal cord position, vocal cord edge, symmetry and regularity of vocal cord vibration were significantly improved than pre-operation（P<0.01）. The five indexes of GRBAS（Grade, Roughness, Breathiness, Asthenia, Strain） and VHI-10, as well as voice acoustic parameters（Jitter, Shimmer, NHR） post-operation were significantly reduced, while the maximum phonation time（MPT） was significantly longer（P<0.01）. The results of laryngeal EMG indicated that the maximum voluntary motor unit recruitment（VMUR） post-operation was significantly recovered（P<0.01）, which confirmed that the affected laryngeal muscle obtained effective nerve reinnervation. Conclusion:Ansa cervicalis anterior root-RLN anastomosis can effectively improve the voice function of patients which is safe and satisfactory. It is an ideal method for the treatment of unilateral RLN injury.

Long noncoding RNA MALAT1 as a ceRNA drives mouse fibroblast activation via the miR-335-3p/P2ry2 axis.

Fibrosis is a complex pathological process that can lead to the permanent loss of biological function, with P2ry2 playing a crucial role in this process. Long non-coding RNAs (lncRNAs) have been reported to play an critically important role in the fibrotic process. However, it remains unclear whether lncRNAs can regulate fibrosis through P2ry2. In this study, we detected the expression of the long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (lnc-MALAT1). We investigated the expression patterns of lnc-MALAT1 and P2ry2 in denervated skeletal muscle, a classical model of fibrosis. Additionally, we utilized a TGF-ß-mediated fibrosis model in NIH/3T3 cells to examine the effects of lnc-MALAT1 and P2ry2 on fibroblast activation and the underlying regulatory mechanisms in vitro. Our results demonstrated that the expression levels of lnc-MALAT1 and P2ry2 were consistently elevated in denervated skeletal muscle, correlating with the degree of fibrosis. In vitro experiments confirmed the regulatory effect of lnc-MALAT1 on P2ry2. Furthermore, we identified miR-335-3p as a potential key molecule in the regulatory relationship of lnc-MALAT1/P2ry2. Dual luciferase reporter assays and AGO2-RIP verified the molecular sponging effect of lnc-MALAT1 on miR-335-3p. Additionally, we validated the regulation of the lnc-MALAT1/miR-335-3p/P2ry2 axis through experimental approaches. In conclusion, our study identified a crucial role of lnc-MALAT1/miR-335-3p/P2ry2 axis in fibroblast activation, providing a promising treatment option against the fibrosis.

Role of tympanocentesis in the prevention of middle ear barotrauma induced by fast buoyant ascent escape from 200 m underwater.

Introduction: We aimed to study middle ear barotrauma caused by fast compression followed by buoyant ascent escape from 200 m underwater and its effect on the auditory system, and to validate the preventive effect of tympanocentesis on middle ear barotrauma. Methods: Twenty Sprague Dawley rats were divided into two groups: rats in group A underwent a simulated fast buoyant ascent escape from a depth of 200 m, while those in group B underwent tympanocentesis before the procedure described for group A. Ear endoscopy, acoustic conductance, and auditory brainstem response (ABR) tests were conducted before and after the procedure to evaluate the severity of middle ear barotrauma and auditory function in both groups. Additionally, histopathological examination of the middle ear in both groups was conducted to evaluate the severity of middle ear barotrauma by observing submucosal haemorrhage. Results: None of the ears in either group showed any abnormalities before the experiment. In group A, middle ear barotrauma was universally observed after the simulation procedure. The tympanograms of all ears were initially type A and became type B after the procedure. Further, after the simulation, the hearing thresholds at different frequencies (4, 8, 16, 24, and 32 kHz) assessed by ABR significantly increased compared to those before the procedure. In group B, no middle ear barotrauma was observed, and the hearing threshold at each frequency did not change significantly compared with post-puncturing. After dissecting the middle ear, gross pathological observations were consistent with the above results. Microscopically, blood accumulation and submucosal haemorrhage in the middle ear cavity were observed in group A but not in group B. Conclusions: Fast buoyant ascent from 200 m underwater can cause middle ear barotrauma, resulting in hearing loss. Tympanic membrane puncture can effectively prevent middle ear barotrauma caused by the rapid buoyant ascent escape procedure.

Effect of nano-hydroxyapatite suspension on cell proliferation and cycle in human periodontal ligament cells.

The study was aimed to provide insights into the effect of nano-hydroxyapatite (nHA) suspension on cell proliferation and cycle of human periodontal ligament cells, offering the evidence for nHA being used in periodontal therapy. Human periodontal ligament cells (HPDLCs) were cultured in different concentrations of nano-hydroxyapatite/sodium carboxymethyl cellulose (nHA-CMCNa) suspension in vitro. After that, cell proliferation ability was examined by MTT [3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and flow cytometry. MTT assay demonstrated that the Relative Proliferation Rate (RPR) of 0.5% nHA-CMCNa group was significantly higher than other groups (p <0.05), which means that nHA-CMCNa might increase cell proliferation ability. Flow cytometry showed that cells in G1 phase decreased, whilst cells in S phase increased after cultured in nHA-CMCNa suspension for 48 h. The result suggested that part of cells finished G1 phase in advance and get into S phase earlier, which speed up the cell proliferation, nHA-CMCNa suspension had great effect on cell proliferation. The high concentration of nHA-CMCNa could shorten the time in G1 phase, impel part of cells into S phase, and accelerate proliferation rate of HPDLCs.

Pathogenic mechanisms and potential therapeutic targets for Parkinson disease revealed by bioinformatic analysis of necroptosis and immune cell infiltration.

Parkinson disease (PD) is an age-dependent neurodegenerative disease with very high prevalence by age 80 years. Necroptosis is a newly identified form of programmed cell death implicated in neurodegenerative diseases, but has not yet been conclusively associated with PD. This study examined the contributions of necroptosis to PD using bioinformatics analysis. Datasets GSE26927, GSE49036, and GSE54536 from the gene expression omnibus database were analyzed for differentially expressed genes (DEGs). These DEGs were then subjected to gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis to identify associated functions and signaling mechanisms. Necroptosis-related differentially expressed genes (NRDEGs) were then identified by the overlap of DEGs and the necroptosis gene set hsa04217. The STRING database and Cytoscape software were then used to build and visualize a protein-protein interaction network and identify hubs and key functional modules among NRDEGs. In addition, immune cell type abundance was analyzed based on DEGs using ImmuCellAI. The identified DEGs, KEGG pathway enrichment terms, and protein-protein interaction network structures of NRDEGs were validated using an independent dataset (GSE54536). The necroptosis pathway was significantly enriched and activated in PD samples. Thirteen NRDEGs were identified in the GSE26927 and GSE49036 datasets, including receptor interacting serine/threonine kinase 1, CASP8 and FADD like apoptosis regulator, TNFRSF1A associated via death domain, and interleukin 1 beta, of which 6 were validated in the GSE54536 dataset. According to gene ontology and KEGG analyses, these NRDEGs are involved in necroptosis-related processes, apoptosis, B cell receptor signaling pathways, and NOD-like receptor signaling pathways. Analysis of DEGs also revealed significant increases in CD8 + T cell and Tex cell infiltration and significant decreases in B cell and T gamma delta cell infiltration within the PD brain. Necroptosis pathways are active in PD and associated with immune cell infiltration. The factors controlling necroptotic signaling and immune infiltration identified in this study may be valuable diagnostic markers and therapeutic targets for PD.

5-aminolevulinic acid photodynamic therapy for extensive oral leukoplakia with concomitant oral submucous fibrosis: A case report.

BACKGROUND: Oral leukoplakia and oral submucous fibrosis are potentially malignant disorders of the oral cavity, with high rates of recurrence and malignant transformation. Notably, the malignant transformation rate of oral leukoplakia with concomitant oral submucous fibrosis is significantly higher than that of oral submucous fibrosis or oral leukoplakia alone. However, the management of these conditions is not well defined. Photodynamic therapy is a minimally invasive treatment modality that effectively targets oral potentially malignant disorders, such as oral leukoplakia, erythroleucoplakia, and verrucous hyperplasia, with the advantages of being repeatable and leaving no scarring. CASE PRESENTATION: We report the case of a 42-year-old man with concomitant oral leukoplakia and oral submucous fibrosis almost involving the entire right buccal mucosa, who underwent six sessions of topical 5-aminolevulinic acid-mediated photodynamic therapy. RESULTS: Photodynamic therapy successfully eradicated whitish plaques and improved mouth opening without any adverse effects. Although photodynamic therapy failed to completely reverse the pathological changes, grading of epithelial dysplasia did not progress and clinical recurrence was not found during the 10-month follow-up. CONCLUSION: In conclusion, topical 5-aminolevulinic acid-mediated photodynamic therapy appears safe and has excellent clinical efficacy against oral leukoplakia-concomitant oral submucous fibrosis, but long-term follow-up is necessary.

Growth and nitrogen metabolism in Sophora japonica (L.) as affected by salinity under different nitrogen forms.

Sophora japonica is a leguminous tree species native to China. To explore the nitrogen (N) source preference and its impact on stress tolerance, a hydroponic experiment was designed in which S. japonica seedlings were supplied with sole ammonium (NH4+) or sole nitrate (NO3-) nutrition under 75 mM NaCl-induced salt stress. The growth and N metabolism performance were investigated. In the absence of NaCl, plants fed NH4+ showed better root growth than those fed NO3-, but there was no difference in aerial part growth. Salinity inhibited the root growth of NH4+-fed plants and the shoot growth of NO3--fed plants, while the total N accumulation was suppressed under either N form. Specifically, in NH4+-fed plants, salinity significantly increased the net photosynthetic rate, root NH4+ content and root antioxidant enzyme activities. Higher nitrate reductase (NR) activities but lower glutamate synthetase (GS) activities were observed in both leaves and roots. Leaf AMT1.1 and AMT2.1a in NH4+-fed plants positively reacted to salt stress, whereas the expression of four AMTs was reduced or remained unchanged in roots. In contrast, salinity suppressed the net photosynthetic rate, antioxidant enzyme activities, and GS activity in the leaves of NO3--fed plants. Upregulation of NPF1.2, NPF2.11, NPF4.6 and NPF7.3, as well as unaltered NR activity, caused higher NO3- content in the leaves. Moreover, NR and glutamate synthase (GOGAT) activities together with the transcription of most NRTs were promoted by salinity in the roots of NO3--fed plants. Additionally, compared to those treated with NH4+, in response to salinity, NO3--treated seedlings showed more intensive repression of the net photosynthetic rate, chlorophyll content, and both shoot and root growth. Overall, these results suggest that S. japonica plants grew better in NH4+ medium than in NO3- medium, and the different N metabolism responses improved S. japonica tolerance to salinity with NH4+ application. This study provides new insights for understanding the mechanism of salt tolerance, breeding resistant varieties of S. japonica, and developing scientific fertilization management strategies during the seedling cultivation period.

Spatial Patterns and Composition Traits of Soil Microbial Nitrogen-Metabolism Genes in the Robinia pseudoacacia Forests at a Regional Scale.

Microbial-driven processes related to the nitrogen-metabolism (N-metabolism) in soil are critical for ecosystem functioning and stability. There are spatial patterns of microbial-mediated nitrogen processes, but we still lack an overview of the soil N-metabolism genes of single nitrogen-fixing tree species pure forests at a regional scale. Here, we investigated the spatial variation and drivers of microbial N-metabolism genes in the rhizosphere soil of Robinia pseudoacacia on the Loess Plateau by metagenomic technology. We found that the distance-decay of soil N functional gene similarities in Robinia pseudoacacia forests on the Loess Plateau spanning a geographic distance of 230 km was significant (p < 0.001). The gene composition and co-occurrence patterns in the process of soil microbial N-metabolism were very different, and they were mainly driven by soil pH and MAP (mean annual precipitation). The proportion of positive links and edges co-occurrence networks between N functional genes increased with increasing pH, suggesting that increasing pH promoted connections between functional genes. The relative frequencies of N-metabolism pathways were consistent on the Loess Plateau, the abundance of ammonia assimilation pathway was highest, and the abundance of the nitrogen fixation pathway was the lowest; only the abundance of the nitrogen fixation pathway was not significantly different. The bacterial and archaeal communities involved in soil nitrogen metabolism were significantly different. Structural equation modeling showed that decreases in soil pH and MAP mainly affected the increase in nitrogen functional gene abundance through an increase in the diversity of N-metabolism microorganisms. In conclusion, this study provides a baseline for biogeographic studies of soil microbe functional genes.

Isolation, characterization, and genome sequencing of a novel chitin deacetylase producing Bacillus aryabhattai TCI-16.

Chitin deacetylase (CDA) is a chitin degradation enzyme that catalyzes the conversion of chitin to chitosan by the deacetylation of N-acetyl-D-glucosamine residues, playing an important role in the high-value utilization of waste chitin. The shells of shrimp and crab are rich in chitin, and mangroves are usually recognized as an active habitat to shrimp and crab. In the present study, a CDA-producing bacterium, strain TCI-16, was isolated and screened from the mangrove soil. Strain TCI-16 was identified and named as Bacillus aryabhattai TCI-16, and the maximum CDA activity in fermentation broth reached 120.35 ± 2.40 U/mL at 36 h of cultivation. Furthermore, the complete genome analysis of B. aryabhattai TCI-16 revealed the chitin-degrading enzyme system at genetic level, in which a total of 13 putative genes were associated with carbohydrate esterase 4 (CE4) family enzymes, including one gene coding CDA, seven genes encoding polysaccharide deacetylases, and five genes encoding peptidoglycan-N-acetyl glucosamine deacetylases. Amino acid sequence analysis showed that the predicted CDA of B. aryabhattai TCI-16 was composed of 236 amino acid residues with a molecular weight of 27.3 kDa, which possessed a conserved CDA active like the known CDAs. However, the CDA of B. aryabhattai TCI-16 showed low homology (approximately 30%) with other microbial CDAs, and its phylogenetic tree belonged to a separate clade in bacteria, suggesting a high probability in structural novelty. In conclusion, the present study indicated that the novel CDA produced by B. aryabhattai TCI-16 might be a promising option for bioconversion of chitin to the value-added chitosan.

[Relationship between the chronic periodontitis and the depression anxiety psychological factor].

OBJECTIVE: To explore the relationship between the chronic periodontitis (CP) and the depression-anxiety psychological factors. METHODS: Thirty-one patients and 29 age, gender-matched volunteers were enrolled for this study. In order to assess the depression-anxiety psychological index, the subjects filled the questionnaire regarding the demographic and socioeconomic information, the oral hygiene habit, the Center for Epidemiologic Studies Depression Scale (CES-D) and the Self Rating Anxiety Scale(ASA). Calculus index (CI), bleeding on probing (BOP), probing depth (PD), clinical attachment loss (CAL), furcation involvement (FI) and tooth mobility were assessed at 6 sites per tooth of all erupted teeth by a manual periodontal probe. The data were analyzed by the analysis of variance, χ(2) test, and multivariable logistic step wise analysis via the software of SPSS 15.0. RESULTS: The mean CAL of the control group was 0.46 ± 0.16,the mean CAL of the moderate, high, and severe CP group was 2.84 ± 0.12, 3.51 ± 0.34, and 4.71 ± 0.51, respectively, which is significant difference between each other (P<0.01). The depression index of the volunteers, the moderate CP, the high CP, and the severe CP was 30.52 ± 3.73, 35.83 ± 7.76, 37.25 ± 6.16, 37.82 ± 5.94, respectively. The anxiety index among the 4 groups was 26.69 ± 3.55, 37.67 ± 6.31, 32.87 ± 5.54, and 35.94 ± 6.30, respectively. The depression and anxiety indexes of the periodontitis groups were higher than those of the control (P<0.01) while there was no significant difference among the 3 CP groups (P>0.05). The multivariate logistic analysis of the relationship between CP and the depression-anxiety psychological factors showed that the depression psychological factor was B=2.301,OR=9.988 while the optimistic coping style was B=-5.174,OR=0.006 in the equation of the regression. CONCLUSION: The depression psychological factor was related to the progression of CP. In addition, the optimistic coping style could prevent the progression of the CP.

[Therapeutic effect of the combination of traditional Chinese medicine and western medicine on patients with oral submucous fibrosis].

OBJECTIVE: To observe the therapeutic effect of salvia miltiorrhiza and prednisolone on patients with oral submucous fibrosis (OSF). METHODS: A total of 60 medium-term OSF patients and 60 advanced stage OSF patients were randomly divided into the first group (treated with both salvia miltiorrhiza and prednisolone) and the second group (treated with prednisolone alone). The clinical effect was compared between each group after 3-month treatment. RESULTS: Difference was found in the lesion area of the medium-term cases and the advanced stage cases of the first group before the treatment [(10.37+/-3.40) cm2, (19.60+/-3.27) cm2] and after the treatment [(5.90+/-4.10) cm2, (16.33+/-4.02) cm2] (P<0.05). Significant difference was found in the mouth opening before the treatment [(3.41+/-0.77) cm, (1.98+/-0.39) cm] and after the treatment [(3.87+/-0.67) cm, (2.26+/-0.46) cm] (P<0.05) in the first group. There was significant difference in the lesion area and mouth opening of the medium-term cases of the second group before the treatment [(10.87+/-3.18) cm2, (3.57+/-0.75) cm] and after the treatment [(6.70+/-3.75) cm2, (3.97+/-0.69) cm] (P<0.05). No difference in the lesion area and mouth opening of the advanced stage cases of the second group was found (P>0.05). There was difference in the therapeutic efficacy between the first group (70%) and the second group (16.67%) of the advanced stage cases (P<0.05), but not in the clinical effect between the 2 groups of the medium-term cases (P>0.05). The side effect of prednisolone could be reduced while used together with salvia miltiorrhiza. CONCLUSION: There is obvious advantage in treating OSF by the combination of salvia miltiorrhiza and prednisolone.