Cytological and metabolomic analysis of Citrus fruit to elucidate puffing disorder.

Puffiness, a physiological disorder commonly observed during the ripening and post-harvest processes of fruits in Citrus reticulata, significantly affects the quality and shelf-life of citrus fruits. The complex array of factors contributing to puffiness has obscured the current understanding of its mechanistic basis. This study examined the puffing index (PI) of 12 citrus varieties at full ripeness, focusing on the albedo layer as a crucial tissue, and investigated the correlation between cellular structural characteristics, key primary metabolites and PI. The findings revealed that the cell gap difference and the number of lipid droplets were closely linked to PI. Chlorogenic acid, Ferulic acid, D-Galacturonic acid, D-Glucuronic acid, (9Z,11E)-Octadecadienoic acid, and 9(10)-EpOME were identified as pivotal primary metabolites for rind puffing. Determination of lignin, protopectin, cellulose and lipoxygenase content further validated the relationship between cell wall, lipid metabolism and rind puffing. This study furnishes novel insights into the mechanisms underlying puffing disorder.

E/D-Mode GaN Inverter on a 150-mm Si Wafer Based on p-GaN Gate E-Mode HEMT Technology.

AlGaN/GaN E/D-mode GaN inverters are successfully fabricated on a 150-mm Si wafer. P-GaN gate technology is applied to be compatible with the commercial E-mode GaN power device technology platform and a systematic study of E/D-mode GaN inverters has been conducted with detail. The key electrical characters have been analyzed from room temperature (RT) to 200 °C. Small variations of the inverters are observed at different temperatures. The logic swing voltage of 2.91 V and 2.89 V are observed at RT and 200 °C at a supply voltage of 3 V. Correspondingly, low/high input noise margins of 0.78 V/1.67 V and 0.68 V/1.72 V are observed at RT and 200 °C. The inverters also demonstrate small rising edge time of the output signal. The results show great potential for GaN smart power integrated circuit (IC) application.

Analysis of the correlation between malocclusion, bad oral habits, and the caries rate in adolescents.

BACKGROUND: Malocclusion deformity and caries are common oral diseases in adolescents. In order to explore the correlation between malocclusion deformity and caries prevalence in adolescents, this study analyzed the influence of factors such as poor oral habits on caries in adolescents. METHODS: In this study, 1,093 adolescents aged 11 to 14 enrolled in 2-3 schools in Zhangjiakou area from September 2020 to June 2021 were selected as the research subjects. The basic data of these adolescents were collected by questionnaires. Malocclusion and caries were examined by on-site inspection. Their bad oral habits were also investigated. After data collection, the chi-square test, logistic regression, the Mann-Whitney U test, and the Kruskal-Wallis H test were used for multivariate analysis. RESULTS: The results showed that the incidence of malocclusion was 72.66% and the caries rate was 68.8% among adolescents aged 11-14 in this area. The 4 bad oral habits of biting, mandibular protrusion, lateral chewing, and mouth breathing were the influencing factors of malocclusion in adolescents. Bad oral habits, malocclusion, and frequent consumption of sugary beverages were independent risk factors for caries in adolescents, and significantly positively correlated with the caries rate. CONCLUSIONS: Malocclusion and the four bad oral habits were independent risk factors of caries in adolescents.

Evaluation and improvement the safety of total marrow irradiation with helical tomotherapy using repeat failure mode and effects analysis.

BACKGROUND & PURPOSE: Helical tomotherapy has been applied to total marrow irradiation (HT-TMI). Our objective was to apply failure mode and effects analysis (FMEA) two times separated by 1 year to evaluate and improve the safety of HT-TMI. MATERIALS AND METHODS: A multidisciplinary team was created. FMEA consists of 4 main steps: (1) Creation of a process map; (2) Identification of all potential failure mode (FM) in the process; (3) Evaluation of the occurrence (O), detectability (D) and severity of impact (S) of each FM according to a scoring criteria (1-10), with the subsequent calculation of the risk priority number (RPN=O*D*S) and (4) Identification of the feasible and effective quality control (QC) methods for the highest risks. A second FMEA was performed for the high-risk FMs based on the same risk analysis team in 1 year later. RESULTS: A total of 39 subprocesses and 122 FMs were derived. First time RPN ranged from 3 to 264.3. Twenty-five FMs were defined as being high-risk, with the top 5 FMs (first RPN/ second RPN): (1) treatment couch movement failure (264.3/102.8); (2) section plan dose junction error in delivery (236.7/110.4); (3) setup check by megavoltage computed tomography (MVCT) failure (216.8/94.6); (4) patient immobilization error (212.5/90.2) and (5) treatment interruption (204.8/134.2). A total of 20 staff members participated in the study. The second RPN value of the top 5 high-risk FMs were all decreased. CONCLUSION: QC interventions were implemented based on the FMEA results. HT-TMI specific treatment couch tests; the arms immobilization methods and strategy of section plan dose junction in delivery were proved to be effective in the improvement of the safety.

Comparative transcript profiling reveals the mechanism of female sterility associated with seedless Ponkan mandarin (Citrus reticulata Blanco).

Seedlessness is a highly desirable trait in citrus varieties. Sterility is the key determination for seedlessness formation. However, the molecular basis for female sterility in seedless mandarin remains unclear. Thus, a seedless Ponkan mandarin (Citrus reticulata Blanco 'Lipeng No.2'), considered the bud mutation of normal seedy Ponkan, was collected to identify candidate genes involved in seedless variation. Suppression subtractive hybridization (SSH) screened 1091 uniESTs related to seedy and seedless Ponkan (727 singlets and 364 contigs), which mainly governed catalytic activity, transferase activity, and oxygen binding. By using RNA-Seq technology, 106 differentially expressed genes (DEGs) were captured, of which 74 were up-regulated and 32 were down-regulated. Gene Ontology and pathway analysis showed that six DEGs were enriched in the biosynthesis of secondary metabolite, whereas five DEGs were enriched in the signaling of plant hormones. The combined results of SSH and RNA-Seq indicated the importance of amino acid metabolism in seedless Ponkan. Our findings revealed that the mechanism of seedless Ponkan generation may be related to gene regulation, signal cascade, and hormone levels. This study provided a solid foundation for functional gene identification in seedless Ponkan and a good reference for relevant research on molecular mechanisms of female sterility in Ponkan mandarin.

Identification of Genes Involved in the Responses of Tangor (C. reticulata × C. sinensis) to Drought Stress.

Drought is the major abiotic stress with adverse effects on citrus, decreasing the agronomical yield and influencing the fruit quality. In this study, cDNA-amplified fragment length polymorphism (cDNA-AFLP) technique was used to investigate the transcriptional profile changes and identify drought-responsive genes in "Amakusa" tangor (C. reticulata × C. sinensis), a hybrid citrus sensitive to water stress. The 255 out of 6,245 transcript-derived fragments (TDFs) displayed altered expression patterns including (A) induction, (B) repression, (C) upregulation, and (D) downregulation. With BLAST search, the gene products of differentially expressed fragments (DEFs) could be classified into several categories: cellular processes, transcription, transport, metabolism, stress/stimuli response, and developmental processes. Downregulated genes were highly represented by photosynthesis and basic metabolism, while upregulated ones were enriched in genes that were involved in transcription regulation, defense, energy, and transport. Present result also revealed some transient and up- and then downregulated genes such as aquaporin protein and photosystem enzyme. Expression patterns of 17 TDFs among 18 homologous to function-known genes were confirmed by qRT-PCR analysis. The present results revealed potential mechanism of drought tolerance in fruit crop and also provided candidate genes for future experiments in citrus.

Colored light-quality selective plastic films affect anthocyanin content, enzyme activities, and the expression of flavonoid genes in strawberry (Fragaria × ananassa) fruit.

The influence of colored light-quality selective plastic films (red, yellow, green, blue, and white) on the content of anthocyanin, the activities of the related enzymes and the transcripts of the flavonoid gene was studied in developing strawberry fruit. The results indicated that colored films had highly significant effects on the total anthocyanin content (TAC) and proportions of individual anthocyanins. Compared with the white control film, the red and yellow films led to the significant increase of TAC, while the green and blue films caused a decrease of TAC. Colored film treatments also significantly affected the related enzyme activity and the expression of structural genes and transcription factor genes, which suggested that the enhancement of TAC by the red and yellow films might have resulted from the activation of related enzymes and transcription factor genes in the flavonoid pathway. Treatment with red and yellow light-quality selective plastic films might be useful as a supplemental cultivation practice for enhancing the anthocyanin content in developing strawberry fruit.

[Renal sympathetic denervation suppresses ventricular substrate remodeling in a canine high-rate pacing model].

OBJECTIVE: To explore the role of renal sympathetic denervation (RSD) on ventricular substrate remodeling in dogs with pacing-induced heart failure (HF). METHODS: A total of 19 dogs were randomized into 3 groups of sham-operated control (n = 7), right ventricular pacing induction of HF (n = 6) and RSD (n = 6). After 8-week pacing induction of HF. Hemodynamic variables were monitored at baseline and after HF. Masson's trichrome staining, enzyme-linked immunosorbent assay (ELISA) and Western blot were used to measure ventricular interstitial fibrosis, brain natriuretic peptide (BNP), angiotensin II (Ang II), aldosterone and transforming growth factor-beta (TGF-ß). RESULTS: All dogs in HF and HF+RSD groups showed increased left and right ventricular diastolic dimensions [left ventricle: (27.0 ± 2.4) vs (37.0 ± 2.8) mm, P < 0.01 and (30.0 ± 2.5) vs (36.0 ± 2.8) mm, P < 0.05; right ventricle: (11.0 ± 1.5) vs (14.0 ± 1.7) mm, P = 0.03 and (12.0 ± 1.1) vs (14.0 ± 1.2) mm, P < 0.05]. Compared with HF + RSD dogs, HF dogs had higher left ventricular end-diastolic pressure (LVEDP) [(25.0 ± 3.7) vs (3.3 ± 1.6) mmHg, P < 0.01] and more fibrous tissue (left ventricle:24.1% ± 4.8% vs 8.5% ± 1.9%, P < 0.01; right ventricle:17.2% ± 5.2% vs 11.8% ± 3.9%, P < 0.01). The levels of BNP, Ang II, aldosterone and TGF-ß in ventricular tissue increased in HF dogs compared to sham-operated and HF+RSD dogs. CONCLUSION: RSD could suppress ventricular substrate remodeling induced by long-term rapid ventricular pacing.

Effect of renal sympathetic denervation on the progression of paroxysmal atrial fibrillation in canines with long-term intermittent atrial pacing.

AIMS: The aim of the present study was to explore the effect of renal sympathetic denervation (RSD) on the progression of paroxysmal atrial fibrillation (AF) in canines with long-term intermittent atrial pacing. METHODS AND RESULTS: Nineteen beagles were randomly divided into sham-operated group (six dogs), control group (six dogs), and RSD group (seven dogs). Sham-operated group were implanted with pacemakers without pacing; control group were implanted with pacemakers with long-term intermittent atrial pacing; and RSD group underwent catheter-based RSD bilaterally and were simultaneously implanted with pacemakers. Atrial pacing was maintained for 8 h a day and a total of 12 weeks in the control group and RSD group. Echocardiography showed that the left atrial structure and function were significantly improved in the RSD group compared with the control group (P < 0.05). Compared with the control group, the RSD group had fewer incidences of AF and a shorter duration of AF (P < 0.05) after long-term intermittent atrial pacing. In addition to increased atrial effective refractory period (AERP) and AF cycle length, AERP dispersion and P-wave duration and dispersion were significantly decreased in the RSD group compared with the control group (P < 0.05). Atrial morphological evaluation suggested that fibrosis and ultrastructural changes induced by long-term intermittent atrial pacing were markedly suppressed in the RSD dogs compared with controls (P < 0.05). Immunohistochemistry results showed that connexin 43 distribution in RSD mid-myocardial was significantly fewer heterogeneous than that in control mid-myocardial (P < 0.05). CONCLUSION: Renal denervation inhibits the progression of paroxysmal AF, which might be related to the suppression of atrial electrophysiology and structural heterogeneity.

Renal sympathetic denervation suppresses ventricular substrate remodelling in a canine high-rate pacing model.

AIMS: This study sought to assess whether renal sympathetic denervation (RSD) could suppress ventricular substrate remodelling and attenuate heart failure (HF) progression. METHODS AND RESULTS: Nineteen dogs were randomised into three groups - seven sham-operated controls, six with right ventricular pacing to induce HF, and six with RSD followed eight weeks later by pacing induction of HF. Haemodynamic variables were monitored at baseline and after HF. Levels of ventricular interstitial fibrosis, BNP, Ang II, aldosterone and TGF-ß were measured. All the dogs in the HF and HFï¹¢RSD groups showed increased left and right ventricular diastolic dimensions, but the dogs in the HFï¹¢RSD group had a higher left ventricular ejection fraction (LVEF) than the HF dogs (0.42±0.05 vs. 0.35±0.04, p<0.01). Compared with the dogs with HF alone, the HF+RSD dogs had lower left ventricular end-diastolic pressure (LVEDP) (3.3±1.6 vs. 25±3.7 mmHg, p<0.01) and less fibrous tissue. The levels of BNP, Ang II, aldosterone and TGF-ß expression in ventricular tissue were higher in the HF dogs than in the sham-operated and HFï¹¢RSD dogs. CONCLUSIONS: RSD suppressed ventricular substrate remodelling induced by long-term rapid ventricular pacing.

Effects of renal sympathetic denervation on the atrial electrophysiology in dogs with pacing-induced heart failure.

BACKGROUND: Heart failure (HF) and atrial fibrillation (AF) are associated with sympathetic activation. Renal sympathetic denervation (RSD) can suppress AF vulnerability. The impact of RSD on atrial electrophysiology in experimental HF is unclear. METHODS: Twenty-two beagles were randomized into control, HF, and HF + RSD groups. Control dogs were implanted cardiac pacemakers without pacing. Dogs in the HF group underwent right ventricular pacing for 3 weeks at 240 beats/min to induce HF. The dogs in the HF + RSD group received RSD and underwent the same HF-inducing procedure. RESULTS: The P-wave dispersion was higher in HF dogs than in the control and HF + RSD dogs (19 ± 3.1 ms vs 13 ± 2.3 ms, 15 ± 2.9 ms, P = 0.04). Conduction time within the interatrium was significantly longer in the HF dogs than that in the control and HF + RSD dogs (39 ± 4 ms vs 31 ± 3 ms, 33 ± 4 ms; P = 0.03). Window of vulnerability (WOV) of AF was widened in the HF dogs than in the HF + RSD dogs (37 ± 5 ms vs 14 ± 3 ms; P < 0.01), while AF could not be induced (WOV = 0) in the control dogs during S1 S2 stimulation. The voltage in the threshold for AF inducibility was lower during ganglionated plexi stimulation in the HF dogs than in the control and HF + RSD dogs (1.8 ± 0.6 V vs 2.5 ± 0.6 V, 2.4 ± 0.4 V; P = 0.04). CONCLUSIONS: RSD could reverse the atrial electrical remodeling and decrease AF inducibility in dogs with pacing-induced HF.

Wenxin Keli suppresses atrial substrate remodelling after epicardial ganglionic plexi ablation.

BACKGROUND: The chronic effects of ganglionic plexi (GP) ablation on atrial fibrillation (AF) inducibility have not been elucidated. OBJECTIVE: To investigate the effect of Wenxin Keli (WK) on the inducibility of AF and atrial substrate remodelling after epicardial GP ablation. METHODS: Twenty dogs were randomly divided into a sham-operated group, a GP ablation group and a WK-treated group. All animals underwent a left thoracotomy at the fourth intercostal space. AF inducibility was assessed by burst rapid pacing at the right atrium. Both the GP ablation group and the WK-treated group received four major GP ablations. In the WK-treated group, dogs were treated with oral WK once per day, and all animals were allowed to recover for eight weeks, after which AF inducibility and AF duration were measured again. RESULTS: After eight weeks of WK treatment, AF inducibility was lower than in the GP ablation group, and was similar to that of the sham-operated group. Compared with the sham-operated group, the levels of atrial natriuretic peptide (ANP), tumour necrosis factor-alpha (TNF-α) and interleukin (IL)-6 in right atrial tissues were increased in GP ablation group (143.6±33.7 pg/mg versus 206.2±41.4 pg/mg, P=0.02; 75.3±12.1 pg/mg versus 141.3±64 pg/mg, P=0.03; and 175.1±42.5 pg/mg versus 351.7±101 pg/mg, P<0.01, respectively). There were no significant differences in levels of ANP, TNF-α and IL-6 in atrial tissues between the sham-operated group and WK treated group. Expression of connexin 43 in atrial tissues was increased after eight weeks of GP ablation, while WK administration inhibited connexin 43 remodelling. CONCLUSIONS: Epicardial GP ablation can induce atrial substrate remodelling, including Cx43 upregulation and increased levels of ANP, TNF-α and IL-6. These changes may be suppressed by long-term oral WK administration.

Effect of renal sympathetic denervation on atrial substrate remodeling in ambulatory canines with prolonged atrial pacing.

We have previously demonstrated that catheter-based renal sympathetic denervation (RSD) could suppress atrial fibrillation (AF) in canines with short-time rapid right atrial pacing (RAP). However, the role of renal denervation on atrial remodeling is unclear. The aim of the present study was to explore the long-term effect of RSD on the atrial remodeling during prolonged RAP. Twenty mongrel dogs were implanted with a high-frequency cardiac pacemaker with a transvenous lead inserted into the right atrial appendage. The dogs were divided into three groups: a sham-operated group (n = 6), the chronic RAP (CRAP) group (n = 7), and the CRAP+RSD group (n = 7). In the CRAP+RSD group, a pacemaker was implanted 6 weeks after RSD was performed bilaterally for recovery. RAP was maintained for 5 weeks in CRAP group and CRAP+RSD group. The plasma levels of Angiotensin II and aldosterone were significantly increased in CRAP group compared with sham-operated group, but the increasing trend was inhibited in CRAP+RSD group compared with CRAP group (P<0.05). Similarly, RSD suppressed the increasing trend that prolonged RAP produced in the left atrial levels of ANP, TNF-α and IL-6. Compared with the sham-operated group, the CRAP group had significantly increased levels of caspase-3, bax and Cx40 whereas the level of Bcl-2 decreased (P<0.05). RSD markedly reduced the upregulation of caspase-3, bax and Cx40 and the downregulation of Bcl-2 expression compared with the CRAP group (P<0.05). Picric acid-sirius red staining study suggested that RSD could markedly alleviate the lesion degree of cardic fibrosis induced by CRAP (P<0.05). Immunohistochemistry results showed that the densities of TH- and GAP43- positive nerves were significantly elevated in the CRAP group compared with the sham-operated group, while RSD operation signicantly inhibited the these changes produced by CRAP. These findings suggest that renal denervation could suppress the atrial remodeling after prolonged RAP in ambulatory canines.

Effect of renal sympathetic denervation on the inducibility of atrial fibrillation during rapid atrial pacing.

BACKGROUND: Atrial fibrillation (AF) is associated with activity of renin-angiotensin-aldosterone system (RAAS). Reduction in renal noradrenaline spillover could be achieved after renal sympathetic denervation (RSD). The relationship between RSD and AF is unknown. OBJECTIVE: The objective of the study was to investigate the inducibility of AF during atrial rapid pacing after RSD. METHODS: Thirteen dogs were used for the study as follows: control group (seven dogs) and RSD group (six dogs). In the control group, dogs were subjected to atrial pacing at 800 beats/min for 7 h, and atrial effective refractory period (AERP) was measured every hour in the status of non-pacing. Subsequently, pacing was stopped and the burst pacing (500 bpm) was repeated to induce AF three times. In the RSD group, after each renal artery ablation, the procedure of pacing and electrophysiological measurement was exactly same as in the control group. Blood was collected before and after pacing to measure the levels of renin, angiotensin II and aldosterone. RESULTS: There was a persistent decrease in AERP in both groups. However, 7 h after cessation of pacing, the induced number of times and duration of AF were higher in the control group than that in the RSD group (1.0 ± 1.26 vs 3.14 ± 2.54, P = 0.03; 16.5 ± 25.1 vs 86.6 ± 116.4, P = 0.02). The plasma aldosterone concentration increased significantly 7 h after rapid pacing in control group (renin, 119.8 ± 31.1 vs 185.3 ± 103.5 pg/ml, P < 0.01; aldosterone, 288.2 ± 43.1 vs 369.6 ± 109.8 pg/ml, P = 0.01). The levels of renin and aldosterone showed a decreasing trend in RSD group, but this did not attain statistical significance. CONCLUSIONS: Episodes of AF could be decreased by renal sympathetic denervation during short-time rapid atrial pacing. This effect might have relationship with decreased activity of RAAS.

[Effects of renal sympathetic denervation on inducibility of atrial fibrillation during rapid atrial pacing].

OBJECTIVE: To explore the effects of renal sympathetic denervation on inducibility of atrial fibrillation (AF) during rapid atrial pacing. METHODS: Thirteen dogs were selected and divided into control group (n=7) and renal artery ablation group (RAA) (n=6). In the control group, the animals were subjected to atrial pacing at 800 beats/min for 7 hours. And atrial effective refractory period (AERP) and induced AF were measured hourly during non-pacing. In the RAA group, after each renal artery ablation, the procedures of pacing and electrophysiological measurement were nearly the same as those in the control group. Blood was collected before and after pacing to measure the levels of rennin, angiotensin AngII (AngII) and aldosterone. RESULTS: There was a persistent decrease in AERP in both groups. However, after a 7-hour pacing, induced number of times and duration of AF were higher in the control group than those in the RAA group. The plasma concentrations of rennin and aldosterone increased significantly after 7-hour rapid pacing in the control group (rennin: (120±31) to (185±104) pg/ml, P<0.01, aldosterone: (288±43) to (370±110) pg/ml, P=0.01). No significant difference existed in the levels of AngII at pre- and post-pacing in the control group ((160±48) to (189±164) pg/ml, P=0.23). The levels of rennin and aldosterone showed a decreasing trend in the RAA group. But there was no statistical significance. CONCLUSIONS: Episodes of AF during short-time rapid atrial pacing may be decreased by renal sympathetic denervation. This effect is probably related with the decreased activity of RAAS.