Genome-wide identification and expression analysis of the BZR gene family in Zanthoxylum armatum DC and functional analysis of ZaBZR1 in drought tolerance.

MAIN CONCLUSION: In this study, six ZaBZRs were identified in Zanthoxylum armatum DC, and all the ZaBZRs were upregulated by abscisic acid (ABA) and drought. Overexpression of ZaBZR1 enhanced the drought tolerance of transgenic Nicotiana benthamian. Brassinosteroids (BRs) are a pivotal class of sterol hormones in plants that play a crucial role in plant growth and development. BZR (brassinazole resistant) is a crucial transcription factor in the signal transduction pathway of BRs. However, the BZR gene family members have not yet been identified in Zanthoxylum armatum DC. In this study, six members of the ZaBZR family were identified by bioinformatic methods. All six ZaBZRs exhibited multiple phosphorylation sites. Phylogenetic and collinearity analyses revealed a closest relationship between ZaBZRs and ZbBZRs located on the B subgenomes. Expression analysis revealed tissue-specific expression patterns of ZaBZRs in Z. armatum, and their promoter regions contained cis-acting elements associated with hormone response and stress induction. Additionally, all six ZaBZRs showed upregulation upon treatment after abscisic acid (ABA) and polyethylene glycol (PEG), indicating their participation in drought response. Subsequently, we conducted an extensive investigation of ZaBZR1. ZaBZR1 showed the highest expression in the root, followed by the stem and terminal bud. Subcellular localization analysis revealed that ZaBZR1 is present in the cytoplasm and nucleus. Overexpression of ZaBZR1 in transgenic Nicotiana benthamiana improved seed germination rate and root growth under drought conditions, reducing water loss rates compared to wild-type plants. Furthermore, ZaBZR1 increased proline content (PRO) and decreased malondialdehyde content (MDA), indicating improved tolerance to drought-induced oxidative stress. The transgenic plants also showed a reduced accumulation of reactive oxygen species. Importantly, ZaBZR1 up-regulated the expression of drought-related genes such as NbP5CS1, NbDREB2A, and NbWRKY44. These findings highlight the potential of ZaBZR1 as a candidate gene for enhancing drought resistance in transgenic N. benthamiana and provide insight into the function of ZaBZRs in Z. armatum.

Gut microbial signatures of patients with diarrhea-predominant irritable bowel syndrome and their healthy relatives.

AIMS: Irritable bowel syndrome (IBS) is a prevalent gastrointestinal disorder, encompassing diarrhea-predominant irritable bowel syndrome (IBS-D). Here, we utilized 16S rDNA gene sequencing to identify potential microbial drivers of IBS-D. METHODS AND RESULTS: A total of 30 healthy relatives and 27 patients with IBS-D were recruited. Clinical data and fecal samples were collected from patients and controls. 16S rDNA gene sequencing was performed to obtain fecal bacterial data. Differences in community composition were evaluated utilizing analysis of similarity (ANOSIM) using Bray-Curtis dissimilarity. The Wilcoxon rank sum test was used to compare differences in taxa and functional pathways. Finally, the key gut microbiota was identified using the random forest algorithm. Gut microbiota diversity, estimated through the Observe, Chao1, and abundance-based coverage estimator (ACE) indices, was significantly lower in the IBS-D patients than in the healthy relatives. ANOSIM analysis further confirmed significant differences in the composition of the gut microbiota between IBS-D patients and healthy relatives, with an R value of 0.106 and a P-value of 0.005. Notably, the IBS-D patients exhibited a significant enrichment of specific bacterial genera, including Fusicatenibacter, Streptococcus, and Klebsiella, which may possess potential pathogenic properties. In particular, the bacterial genus Klebsiella demonstrated a positive correlation with irritable bowel syndrome severity scoring system scores. Conversely, healthy subjects showed enrichment of bacterial genera such as Alistipes, Akkermansia, and Dialister, which may be beneficial bacteria in IBS-D. Utilizing the random forest model, we developed a discriminative model for IBS-D based on differential bacterial genera. This model exhibited impressive performance, with an area under the curve value of 0.90. Additionally, our analysis did not reveal any gender-specific differences in the microbiota community composition among IBS-D patients. CONCLUSIONS: Our findings offer preliminary insights into the potential relationship between intestinal microbiota and IBS-D. The identification model for IBS-D, grounded in gut microbiota, holds promising prospects for improving early diagnosis of IBS-D.

Characterization and Prebiotic Potential of Longan Juice Obtained by Enzymatic Conversion of Constituent Sucrose into Fructo-Oligosaccharides.

The prebiotic potential of longan juice obtained by a commercial Viscozyme L for conversion of constituent sucrose to fructo-oligosaccharide was investigated. The physicochemical properties and carbohydrate composition of the longan juice was evaluated before and after enzymatic treatment. The stimulation effects of the treated longan juice on probiotic bacteria growth were also studied in vitro. The results showed that total soluble solids, yield and clarity of longan juice were all significantly improved after enzyme treatment. The water-soluble polysaccharide content, including pectin, was significantly increased. Compared with the natural longan pulp, the enzyme treated juice showed a significant decrease in sucrose content. Substantial fructo-oligosaccharides including 1-kestose and nystose were synthesized after enzyme treatment. The molecular weight distribution and the monosaccharide composition of the water-soluble polysaccharide were significantly changed by enzyme treatment. The treated longan juice and its ethanol-soluble sugar fraction promoted the growth of Streptococus thermophiles, Lactobacillus acidophilus and Lactobacillus delbrueckii, showing a good potential of the treated longan juice for producing functional foods and nutraceuticals.

A Zinc Finger Protein Regulates Flowering Time and Abiotic Stress Tolerance in Chrysanthemum by Modulating Gibberellin Biosynthesis.

Flowering time and an ability to tolerate abiotic stresses are important for plant growth and development. We characterized BBX24, a zinc finger transcription factor gene, from Chrysanthemum morifolium and found it to be associated with both flowering time and stress tolerance. Transgenic lines with suppressed expression of Cm-BBX24 (Cm-BBX24-RNAi) flowered earlier than wild-type plants and showed decreased tolerance to freezing and drought stresses. Global expression analysis revealed that genes associated with both photoperiod and gibberellin (GA) biosynthesis pathways were upregulated in Cm-BBX24-RNAi lines, relative to the wild type. By contrast, genes that were upregulated in overexpressing lines (Cm-BBX24-OX), but downregulated in Cm-BBX24-RNAi lines (both relative to the wild type), included genes related to compatible solutes and carbohydrate metabolism, both of which are associated with abiotic stress. Cm-BBX24 expression was also influenced by daylength and GA4/7 application. Under long days, changes in endogenous GA1, GA4, GA19, and GA20 levels occurred in young leaves of transgenic lines, relative to the wild type. Regulation of flowering involves the FLOWERING TIME gene, which integrates photoperiod and GA biosynthesis pathways. We postulate that Cm-BBX24 plays a dual role, modulating both flowering time and abiotic stress tolerance in chrysanthemum, at least in part by influencing GA biosynthesis.

Effects of simulated digestion on the structural characteristics and dendritic cell activation of longan polysaccharides.

An active polysaccharide (LP) from longan was purified and characterized. LP consisted of galactose and glucose in a molar ratio of 1.5: 98.5, with a molecular weight of 4.67 × 107 g/mol. The main backbone of LP was T-α-D-Glcp-[(1 â†’ 6)-α-D-Glcp-(1 â†’ 6)-α-D-Glcp]n. After simulated gastrointestinal digestion, the molecular weight distribution, monosaccharide composition, and major glycosidic bonds of LP were not significantly changed. LP and digested LP (DLP) reduced phagocytosis and promoted IL-10 and IL-12 secretion of dendritic cells. In addition, the effects of LP and DLP on activating dendritic cells showed no significant difference. This study helps to illuminate the potential mode of immunomodulatory action of longan polysaccharides in vivo.

Acetylation at the O-6 position of t-Glc improved immunoactivity of α-1,6-glucan from longan by additionally activating Dectin-1 and CD14 receptors.

Acetylation is an important approach to improve the bioactivity of polysaccharides; however, the mechanisms have not been fully understood. As a key component of longan for exerting health promoting function, longan polysaccharide was hypothesized may achieve elevated immunoregulatory activity after acetylation. A bioactive longan polysaccharide (LP) composed of (1 â†’ 6)-α-d-glucan (84.1 %) and with an average Mw of 9.68 × 104 kDa was acetylated to different degree of substitutions (DS) in this study. Key structural changes responsible for improvement in immunoregulatory activity were identified, and underlying mechanisms were investigated. Acetylated LP (Ac-LP) with DS 0.37, 0.78 and 0.92 were obtained. Structural characterization identified the substitution of acetyl groups occurs at O-6 positions of t-Glc non-selectively, while the backbone structure was not apparently changed. This resulted in increased expression of cytokines (IL-10, IL-6 and TNF-α) and ROS production in RAW264.7 macrophages, indicating improved immune activity which is positively related to the DS of Ac-LP. This is attribute to additional cellular receptors for Ac-LP (CD14 and Dectin-1) apart from receptors for LP (TLR4 and Ca2+ receptors), as well as the relative higher protein expression of TLR4-MyD88 signaling pathways. These results would provide guidance for the utilization of acetylated polysaccharides with improved immunoactivity.

A Novel Polysaccharide Isolated From Fresh Longan (Dimocarpus longan Lour.) Activates Macrophage via TLR2/4-Mediated PI3/AKT and MyD88/TRAF6 Pathways.

A novel immunomodulatory polysaccharide (LP4) with a molecular weight 6.31 × 104 g/mol was purified from fresh longan pulp. It was composed of mannose, glucose, glucuronic acid, galactose, xylose, arabinose, galacturonic acid, fucose, and rhamnose in a molar percentage of 36:31:10:7:4:4:3:2:2, and mainly linked by (1→6)-ß-Man, (1→4)-ß-Glc and (1→6)-α-Glc. LP4 can obviously enhance the phagocytosis of macrophages and promote the proliferation of lymphocytes. After treating macrophages with LP4 (12.5-50 µg/ml), the production of IL-1ß and TNF-α was significantly increased. These increases of cytokines were suppressed when the TLR2/TLR4 receptors were inhibited by anti-TLR2 and/or anti-TLR4 antibodies. Moreover, the mRNA expression of INOS, AKT, PI3K, TRAF6 and MyD88 was significantly suppressed by TLR2/TLR4 antibodies. These results indicated that LP4 induced macrophage activation mainly via the TLR2 and TLR4-induced PI3K/AKT and MyD88/TRAF6 pathways.

In vitro immunomodulatory activity of water-soluble glucans from fresh and dried Longan (Dimocarpus longan Lour.).

Longan (Dimocarpus longan Lour.) is a seasonal tropical fruit used by Chinese medicine in both fresh and dried pulp forms. Their polysaccharides have been reported to have biological activity. However, their composition and immune activity have not yet been disclosed. To fulfil this aim, hot water-soluble polysaccharides of fresh and dried longan pulp were fractionated according to their molecular weight by ultrafiltration (10, 50, 100 kDa cut off). The main polysaccharides recovered were 1,6-linked glucans branched at O-3 (4-8%), O-2 (1%), O-2,4 (0.1%), and O-3,4 (0.1%). The drying process promotes the solubility of the polysaccharides. These glucans from fresh and dried longan pulp have immunomodulatory activity, shown by in vitro phagocytosis, NO, TNF-α, and IL-6 macrophages production assays. They showed also to inhibit the inflammatory response induced by LPS. The immunological activity of these glucans seems to have different responses dependent on their molecular weight, related to the immune regulatory pathways.

Glucose-rich polysaccharide from dried 'Shixia' longan activates macrophages through Ca2+ and CR3- mediated MAPKs and PI3K-AKT pathways.

A water-soluble glucose-rich polysaccharide from dried 'Shixia' longan pulp (LPsx) has been isolated for the first time, and its structure and immuno-regulatory mechanism were studied. LPsx is a hetero-polysaccharide with the average molecular weight 4102 g/mol. It was mainly consisted of glucose (95.9%), and small proportions of arabinose (2.1%), galactose (1.0%), mannose (0.6%), and xylose (0.4%). As analyzed by NMR, LPsx was mainly composed of (1 â†’ 6)-α-d-glucose and (1 â†’ 6)-ß-d-glucose, branched with α-d-glucose-(1→. The immunomodulatory activity study showed that LPsx significantly increased the phagocytosis of macrophages, and strongly promoted the production of NO, IL-1ß, IL-6 and TNF-α. Moreover, LPsx could inhibit the inflammatory response induced by lipopolysaccharide. The immuno-regulatory mechanism of LPsx was studied using RNA- sequencing and receptors activity analyses. It was found that LPsx induced macrophage activation via Ca2+ and CR3-mediated MAPKs and PI3K-AKT signaling pathways. The results would be helpful for revealing the health promoting mechanism of dried 'Shixia' longan in traditional Chinese medicine.

[Facial Demodex infection among college students in Tangshan].

A survey indicated that the prevalence of Demodex infection among 512 college students in Tangshan was 36.3% (186/512), that of males and females was 39.3% (81/206) and 34.3% (105/306) respectively (P>0.05). The infection of Demodex folliculorum accounted for 82.3% (153/186), followed by D. brevis (7.5%, 14/186) and mixed infection (10.2%, 19/186). The prevalence was 47.0% (93/198) in subjects with oily skin, 26.6% (37/139) in those with dry skin, and 33.9% (56/165) in mixed-type skin (P<0.05). Subjects with facial diseases (62.0%, 75/121), such as rosacea and acne, were more likely to be infected with Demodex than those with healthy skin (27.6%, 80/290) (P<0.05). Prevalence in those lived in humid environment (67.9%, 95/140) was higher than those lived in the desiccating environment (24.5%, 91/372) (P<0.05).

[Study of subtotal colectomy combined with psychological intervention in treating slow transit constipation].

OBJECTIVE: To investigate the value of psychological intervention in treating slow transit constipation (STC), and to provide the reference to clinical treatment for STC patients with psychological disorder. METHODS: A total of 94 STC patients with psychological disorder admitted to the Anorectal Hospital of Chengdu from June 2010 to August 2012 were prospectively enrolled and divided into psychological intervention group(subtotal colectomy plus postoperative psychological intervention) and control group (subtotal colectomy without postoperative psychological intervention). Scores of Hamilton depression scale (HAMD), Hamilton anxiety scale(HAMA), Wexner constipation scale (WCS) and gastrointestinal quality-of-life index(GIQLI) were recorded 1, 3, 6, 12 and 24 months after operation. SPSS 17.0 statistical software was used to analyze the data. RESULTS: There were no differences in baseline data, operative time, blood loss, time to the first flatus and time to the first defecation between two groups(all P>0.05). The scores of HAMD and HAMA were significantly reduced in psychological intervention group compared with control group 3, 6, 12 and 24 months after operation (all P<0.05). Ratios of cure, obvious progress, progress and invalidation of depression symptoms in psychological intervention group were 2.6%(1/39), 66.7%(26/39), 25.6%(10/39) and 5.1% (2/39) respectively at postoperative 24-month, which were better than those [0, 34.2%(13/38), 44.7% (17/38) and 21.1%(8/38) respectively] in control group with significant difference(P=0.013). Ratios of cure, obvious progress, progress and invalidation of anxiety symptoms in psychological intervention group were 10.3%(4/39), 53.8%(21/39), 28.2%(11/39) and 7.7%(3/39) respectively at postoperative 24-month, which were better than those [0, 28.9%(11/38), 55.3%(21/38) and 15.8%(6/38) respectively] in control group with significant difference (P=0.011). The WCS scores in psychological intervention group were lower than those in control group 6, 12, 24 months after operation (all P<0.05). The GIQLI scores in psychological intervention group were higher than those in control group 3, 6, 12 and 24 months after operation(all P<0.05). CONCLUSION: Subtotal colectomy combined with psychological intervention not only can significantly improve psychological disorder, but also increase the efficacy of surgery in the treatment of slow transit constipation patients with psychological disorder.

[Self-control study of dynamic multiple pelvic angiography and pelvic four-contrast defecography in the diagnosis of functional defecation disorder].

OBJECTIVE: To evaluate and compare the value of dynamic multiple pelvic angiography and pelvic four-contrast defecography in the diagnosis of functional defecation disorder. METHODS: From September 2014 to July 2015, a prospective controlled trial was carried out in Chengdu Anorectal Hospital. A total of 32 patients met the inclusion criteria of functional defecation disorder simultaneously underwent pelvic four-contrast defecography and dynamic multiple pelvic angiography. The diagnostic results of these two methods were compared. RESULTS: The absolute values of anorectal angle and level of perineum, peritoneum and bladder from rest to defecation were (29.6±13.6)°, (26.2±14.2) mm, (55.5±25.6) mm and (28.9±16.5) mm in dynamic multiple pelvic angiography, and (24.6±5.8)° (18.7±10.6) mm, (34.5±18.4) mm and (19.2±11.8) mm in pelvic four-contrast defecography respectively, whose differences were statistically significant (P = 0.026, 0.022, 0.000, 0.011 respectively). The diagnostic rate of pelvic peritoneal hernia was 93.8%(30/32) and 68.8%(22/32) in dynamic multiple pelvic angiography and pelvic four-contrast defecography respectively with significant difference(P=0.011). CONCLUSION: Dynamic multiple pelvic angiography has significant advantage in the diagnosis of pelvic peritoneal hernia, and can provide a more objective basis for the diagnosis of functional defecation disorder.

[Laparoscopy-assisted subtotal colectomy with transanal specimen extraction for slow transit constipation].

OBJECTIVE: To investigate the clinical application of laparoscopy-assisted subtotal colectomy with transanal specimen extraction for slow transit constipation(STC). METHODS: Retrospective analysis was performed on the clinical data of 8 cases with STC undergoing the procedure mentioned above from February to November 2013. Pre-and post-operative constipation was assessed using Wexner Constipation and Incontinence Scales, and quality of life was assessed using Gastrointestinal Quality of Life Index. RESULTS: All the operations were completely successful without postoperative complications, such as intestinal fistula, pelvic infection, anastomotic stricture, intestinal obstruction. The Operative time was (287.6 ± 21.5) min, blood loss was (109.7 ± 41.1) ml, time to first flatus was (2.5 ± 0.9) d. The proportion of postoperative constipation symptom index improvement was(77.6 ± 8.3)%. Postoperative quality of life score was 97.3 ± 15.7, significantly higher than that before operation(P<0.05). Postoperative Wexner constipation score was 8.8 ± 3.7, significantly lower than that before operation. CONCLUSION: Laparoscopy-assisted subtotal colectomy with transanal specimen extraction in the treatment of STC has good short-term efficacy with obvious improvement in quality of life.