Two-Dimensional Biodegradable Black Phosphorus Nanosheets Promote Large Full-Thickness Wound Healing through In Situ Regeneration Therapy.

Large full-thickness skin lesions have been one of the most challenging clinical problems in plastic surgery repair and reconstruction. To achieve in situ skin regeneration and perfect clinical outcomes, we must address two significant obstacles: angiogenesis deficiency and inflammatory dysfunction. Recently, black phosphorus has shown great promise in wound healing. However, few studies have explored the bio-effects of BP to promote in situ skin regeneration based on its nanoproperties. Here, to investigate whether black phosphorus nanosheets have positive bio-effects on in situ skin repair, we verified black phosphorus nanosheets' positive effects on angiogenic and anti-inflammatory abilities in vitro. Next, the in vivo evaluation performed on the rat large full-thickness excisional wound splinting model more comprehensively showed that the positive bio-effects of black phosphorus nanosheets are multilevel in wound healing, which can effectively enhance anti-inflammatory ability, angiogenesis, collagen deposition, and skin re-epithelialization. Then, multiomics analysis was performed to explore further the mechanism of black phosphorus nanosheets' regulation of endothelial cells in depth. Molecular mechanistically, black phosphorus nanosheets activated the JAK-STAT-OAS signaling pathway to promote cellular function and mitochondrial energy metabolism in endothelial cells. This study can provide a theoretical basis for applying two-dimensional black phosphorus nanosheets as nanomedicine to achieve in situ tissue regeneration in complex human pathological microenvironments, guiding the subsequent optimization of black phosphorus.

The Impact of Esketamine Combined with Dexmedetomidine on Laparoscopic Gallbladder Surgery: A Randomized Controlled Trial.

Objective: To assess the efficacy of combining esketamine with dexmedetomidine in laparoscopic gallbladder surgery. Methods: We investigated 110 laparoscopic cholecystectomy patients at Jinan Central Hospital, affiliated with Shandong First Medical University, from April 2019 to March 2020. Patients were randomly assigned to the control group (n = 55) or observation group (n = 55). The control group received dexmedetomidine intravenously at 1 µg/kg and a continuous infusion at 0.5 µg•kg-1•h-1. The observation group received esketamine and dexmedetomidine, with intravenous esketamine at 0.4 mg/kg and a continuous infusion at 0.1 mg/(kg•h). We measured heart rate (HR), systolic blood pressure (SBP), and diastolic blood pressure (DBP) at four-time points: before anesthesia (T0), 30 minutes after anesthesia (T1), extubation (T2), and awakening (T3). We also assessed wake time, post-anesthesia care unit (PACU) stay, and Ramasy and visual analogue scale (VAS) scores at 2, 6, 12, and 24 hours post-surgery. Results: At T0, no significant changes occurred in HR, SBP, and DBP in both groups (P > .05). However, at T1 and T2, HR, SBP, and DBP gradually decreased, with the control group exhibiting lower levels than the observation group (P < .05). These levels returned to baseline at T3. PACU residence and wake times showed no significant differences (P > .05). At 2 hours post-operation, Ramasy scores significantly dropped in the observation group versus the control group (P < .05). At 6, 12, and 24 hours post-operation, Ramasy scores exhibited no significant differences (P > .05). Moreover, at 2, 6, and 12 hours post-operation, VAS scores in the observation group were notably lower than in the control group (P < .05). At 24 hours post-operation, VAS scores revealed no significant differences (P > .05). Adverse reactions within 3 days post-operation did not differ significantly between the groups (P > .05). Conclusions: Combining esketamine with dexmedetomidine enhances the quality of laparoscopic cholecystectomy, alleviates postoperative agitation, accelerates cognitive function recovery, reduces cognitive function impairment, and merits clinical consideration.

Deep learning and targeted metabolomics-based monitoring of chewing insects in tea plants and screening defense compounds.

Tea is an important cash crop that is often consumed by chewing pests, resulting in reduced yields and economic losses. It is important to establish a method to quickly identify the degree of damage to tea plants caused by leaf-eating insects and screen green control compounds. This study was performed through the combination of deep learning and targeted metabolomics, in vitro feeding experiment, enzymic analysis and transient genetic transformation. A small target damage detection model based on YOLOv5 with Transformer Prediction Head (TPH-YOLOv5) algorithm for the tea canopy level was established. Orthogonal partial least squares (OPLS) was used to analyze the correlation between the degree of damage and the phenolic metabolites. A potential defensive compound, (-)-epicatechin-3-O-caffeoate (EC-CA), was screened. In vitro feeding experiments showed that compared with EC and epicatechin gallate, Ectropis grisescens exhibited more significant antifeeding against EC-CA. In vitro enzymatic experiments showed that the hydroxycinnamoyl transferase (CsHCTs) recombinant protein has substrate promiscuity and can catalyze the synthesis of EC-CA. Transient overexpression of CsHCTs in tea leaves effectively reduced the degree of damage to tea leaves. This study provides important reference values and application prospects for the effective monitoring of pests in tea gardens and screening of green chemical control substances.

Effect of dietary supplementation with multinutrient soy flour on body composition and cognitive function in elderly individuals at the risk of low protein: a randomized, double-blind, placebo-controlled study.

Insufficient protein intake and cognitive decline are common in older adults; however, there have been few studies on low protein risk screening and complex nutrient interventions for elderly individuals in rural communities. This study aimed to evaluate the effect of dietary multinutrient soy flour (MNSF) on body composition and cognitive function in elderly individuals who are at risk of protein deficiency in a randomized, double-blind, placebo-controlled clinical trial. Nutritional interventions were given to those found to have low protein levels using bioelectrical impedance analysis (BIA). Among 733 older adults screened, 62 participants were included and randomly assigned into two groups, one taking soy flour and the other taking MNSF for 12 weeks. A previous cross-sectional survey found that 35.1% of the elderly people with an average age of 71.61 ± 5.94 years had an inadequate body protein mass proportion. After the intervention, the MNSF group demonstrated a significant improvement in protein mass, muscle mass, mineral levels, skeletal muscle mass, and fat-free mass compared with baseline (all P < 0.05), as well as a better upward trend compared with the soy flour group (P = 0.08; P = 0.07; P = 0.05; P = 0.08; P = 0.07). Regarding the mini-mental state examination (MMSE) scores, the MNSF group showed a significant decrease after 12 weeks (P < 0.05), which were significantly different compared with the soy flour group (P < 0.05). In the future, the application of MNSF as a food-based supplement to improve nutrition and delay cognitive decline in older adults at the risk of protein deficiency may be considered.

Flavan-3-ol Galloylation-Related Functional Gene Cluster and the Functional Diversification of SCPL Paralogs in Camellia sp.

The high accumulation of galloylated flavan-3-ols in Camellia sp. is a noteworthy phenomenon. We identified a flavan-3-ol galloylation-related functional gene cluster in tannin-rich plant Camellia sp., which included UGT84A22 and SCPL-AT gene clusters. We investigated the possible correlation between the accumulation of metabolites and the expression of SCPL-ATs and UGT84A22. The results revealed that C. sinensis, C. ptilophylla, and C. oleifera accumulated galloylated cis-flavan-3-ols (EGCG), galloylated trans-flavan-3-ols (GCG), and hydrolyzed tannins, respectively; however, C. nitidissima did not accumulate any galloylated compounds. C. nitidissima exhibited no expression of SCPL-AT or UGT84A22, whereas the other three species of Camellia exhibited various expression patterns. This indicated that the functions of the paralogs of SCPL-AT vary. Enzymatic analysis revealed that SCPL5 was neofunctionalized as a noncatalytic chaperone paralog, a type of chaerone-like protein, associating with flavan-3-ol galloylation; moreover, CsSCPL4 was subfunctionalized in association with the galloylation of cis- and trans-flavan-3-ols. In C. nitidissima, an SCPL4 homolog was noted with mutations in two cysteine residues forming a disulfide bond, which suggested that this homolog was defunctionalized. The findings of this study improve our understanding of the functional diversification of SCPL paralogs in Camellia sp.

New insights into the function of plant tannase with promiscuous acyltransferase activity.

Plant tannases (TAs) or tannin acyl hydrolases, a class of recently reported carboxylesterases in tannin-rich plants, are involved in the degalloylation of two important groups of secondary metabolites: flavan-3-ol gallates and hydrolyzable tannins. In this paper, we have made new progress in studying the function of tea (Camellia sinensis) (Cs) TA-it is a hydrolase with promiscuous acyltransferase activity in vitro and in vivo and promotes the synthesis of simple galloyl glucoses and flavan-3-ol gallates in plants. We studied the functions of CsTA through enzyme analysis, protein mass spectrometry, and metabolic analysis of genetically modified plants. Firstly, CsTA was found to be not only a hydrolase but also an acyltransferase. In the two-step catalytic reaction where CsTA hydrolyzes the galloylated compounds epigallocatechin-3-gallate or 1,2,3,4,6-penta-O-galloyl-ß-d-glucose into their degalloylated forms, a long-lived covalently bound Ser159-linked galloyl-enzyme intermediate is also formed. Under nucleophilic attack, the galloyl group on the intermediate is transferred to the nucleophilic acyl acceptor (such as water, methanol, flavan-3-ols, and simple galloyl glucoses). Then, metabolic analysis suggested that transient overexpression of TAs in young strawberry (Fragaria × ananassa) fruits, young leaves of tea plants, and young leaves of Chinese bayberry (Myrica rubra) actually increased the total contents of simple galloyl glucoses and flavan-3-ol gallates. Overall, these findings provide new insights into the promiscuous acyltransferase activity of plant TA.

Flavonol-Aluminum Complex Formation: Enhancing Aluminum Accumulation in Tea Plants.

Polyphenol-rich tea plants are aluminum (Al) accumulators. Whether an association exists between polyphenols and Al accumulation in tea plants remains unclear. This study revealed that the accumulation of the total Al and bound Al contents were both higher in tea samples with high flavonol content than in low, and Al accumulation in tea plants was significantly and positively correlated with their flavonol content. Furthermore, the capability of flavonols combined with Al was higher than that of epigallocatechin gallate (EGCG) and root proanthocyanidins (PAs) under identical conditions. Flavonol-Al complexes signals (94 ppm) were detected in the tender roots and old leaves of tea plants through solid-state 27Al nuclear magnetic resonance (NMR) imaging, and the strength of the signals in the high flavonol content tea samples was considerably stronger than that in the low flavonol content tea samples. This study provides a new perspective for studying Al accumulation in different tea varieties.

Gegen Qinlian decoction ameliorates murine colitis by inhibiting the expansion of Enterobacteriaceae through activating PPAR-γ signaling.

Ulcerative colitis (UC) is a chronic and relapsing inflammatory disease of the intestine. Dysbiosis, especially the expansion of facultative anaerobic Enterobacteriaceae, maybe the main pathogenesis of UC. Gegen Qinlian decoction (GD), a traditional Chinese medicinal formula chronicled in the Shang Han Lun, is commonly used to treat UC and has shown an excellent effect on inducing disease remission. However, the role of GD in regulating gut microbiota has not been fully clarified. Herein, we investigated the potential effect of GD on inhibiting the expansion of Enterobacteriaceae and further explored the potential mechanism of this action. Our study demonstrated that GD remarkably reduced body weight loss of colitis mice, shortening of colon length, and inflammation of the colon. Peroxisome proliferator-activated receptor-γ (PPAR-γ) signaling was inactivated in colitis colon tissue, and the abundance of Escherichia coli (E. coli, family of Enterobacteriaceae) in colonic contents and the concentration of lipopolysaccharide (LPS) in colonic tissue were significantly upregulated after DSS-treatment. Notably, GD administration can result in the activation of PPAR-γ and inactivation of iNOS, which lead to the reduction of nitrate, the inhibition of E. coli, and less production of LPS. Combined GD with PPAR-γ antagonist, the effect of GD on the treatment of UC was weakened, and effectless in inhibiting the expansion of Enterobacteriaceae. Therefore, GD ameliorates UC by preventing a dysbiotic expansion of potentially pathogenic E. coli by reducing nitrate levels in the lumen through activating PPAR-γ signaling.

Curcumae longae Rhizoma (Jianghuang) extract reverses the 5-Fluoruracil resistance in colorectal cancer cells via TLR4/PI3K/Akt/mTOR pathway.

OBJECTIVES: Intensive efforts have been made in the area of identifying drug resistance modulators from traditional Chinese medicine. Various natural plant extracts have been reported for their reversal effect of drug resistance in cancers. This study was to assess the reversal potential of Curcumae longae Rhizoma extract (CLRE) in 5-Fluorouracil (5-Fu) resistance to colon cancer and explore its underly mechanism. METHODS: Increased concentrations of 5-Fu were used to culture SW480. A series of concentrations of CLRE were used to treat the 5-Fu resistant SW480 cells. WST-8 assay was used to detect the cell viability. Cell apoptosis was assessed by SuperView™ 488 Caspase-3 Assay Kit. The quantification of mentioned factors was archived by RT-qPCR. Network pharmacology analysis was used to explore the target of CLRE. RESULTS: 5-Fu resistant cell line (SW480/5-FuR) was established. The IC50 value of CLRE against SW480/5-FuR was 181.0 ± 14.12 µg/ml. CLRE can resensitize the SW480/5-FuR to 5-Fu by inhibiting cell growth. The combination treatment (CLRE+5-Fu) induced cell apoptosis via inhibition of bcl-2 and activation of caspase-3 and bax. Three active ingredients from CLRE were identified. TLR4 was targeted by these three ingredients and linked these ingredients to PI3K/Akt/mTOR pathway. The levels of TLR4, PI3K, AKT1, and mTORC1 mRNA were decreased when 5-Fu was combined with CLRE. CONCLUSIONS: CLRE could reverse 5-Fu resistance in colon cancer by inactivating TLR4/PI3K/AKT/mTORC1 pathway. This finding might provide a molecular basis and a valuable direction for further clinical applications and research for treating 5-Fu resistant colon cancer.

Molecular and biochemical characterization of two 4-coumarate: CoA ligase genes in tea plant (Camellia sinensis).

KEY MESSAGE: Two 4-coumarate: CoA ligase genes in tea plant involved in phenylpropanoids biosynthesis and response to environmental stresses. Tea plant is rich in flavonoids benefiting human health. Lignin is essential for tea plant growth. Both flavonoids and lignin defend plants from stresses. The biosynthesis of lignin and flavonoids shares a key intermediate, 4-coumaroyl-CoA, which is formed from 4-coumaric acid catalyzed by 4-coumaric acid: CoA ligase (4CL). Herein, we report two 4CL paralogs from tea plant, Cs4CL1 and Cs4CL2, which are a member of class I and II of this gene family, respectively. Cs4CL1 was mainly expressed in roots and stems, while Cs4CL2 was mainly expressed in leaves. The promoter of Cs4CL1 had AC, nine types of light sensitive (LSE), four types of stress-inducible (SIE), and two types of meristem-specific elements (MSE). The promoter of Cs4CL2 also had AC and nine types of LSEs, but only had two types of SIEs and did not have MSEs. In addition, the LSEs varied in the two promoters. Based on the different features of regulatory elements, three stress treatments were tested to understand their expression responses to different conditions. The resulting data indicated that the expression of Cs4CL1 was sensitive to mechanical wounding, while the expression of Cs4CL2 was UV-B-inducible. Enzymatic assays showed that both recombinant Cs4CL1 and Cs4CL2 transformed 4-coumaric acid (CM), ferulic acid (FR), and caffeic acid (CF) to their corresponding CoA ethers. Kinetic analysis indicated that the recombinant Cs4CL1 preferred to catalyze CF, while the recombinant Cs4CL2 favored to catalyze CM. The overexpression of both Cs4CL1 and Cs4CL2 increased the levels of chlorogenic acid and total lignin in transgenic tobacco seedlings. In addition, the overexpression of Cs4CL2 consistently increased the levels of three flavonoid compounds. These findings indicate the differences of Cs4CL1 and Cs4CL2 in the phenylpropanoid metabolism.

Two UDP-Glycosyltransferases Catalyze the Biosynthesis of Bitter Flavonoid 7-O-Neohesperidoside through Sequential Glycosylation in Tea Plants.

Flavonoid glycosides are typical bitter and astringent tasting compounds that contribute to the taste of tea beverages. However, the genes that contribute to the biosynthesis of bitter compounds (e.g., flavanone 7-O-neohesperidoside) in tea plants have yet to be identified. In this study, we identified 194 UDP-glycosyltransferases (UGTs) from the tea transcriptome database. Among them, two genes, CsUGT75L12 and CsUGT79B28, encoding flavonoid 7-O-glycosyltransferase and 7-O-glucoside(1→2)rhamnosyltransferase, respectively, were identified from Camellia sinensis. In vitro, the purified recombinant enzyme rCsUGT75L12 specifically transports the glucose unit from UDP-glucose to the 7-OH position of the flavonoid to produce the respective 7-O-glucoside. rCsUGT79B28 regiospecifically transfers a rhamnose unit from UDP-rhamnose to the 2″-OH position of flavonoid 7-O-glucosides to produce flavonoid 7-O-di-glycosides. Additionally, the expression profiles of the two CsUGTs were correlated with the accumulation patterns of 7-O-glucoside and 7-O-neohesperidoside, respectively, in tea plants. These results indicated that the two CsUGTs are involved in the biosynthesis of bitter flavonoid 7-O-neohesperidoside through the sequential glucosylation and rhamnosylation of flavonoids in C. sinensis. Taken together, our findings provided not only molecular insights into flavonoid di-glycoside metabolism in tea plants but also crucial molecular markers for controlling the bitterness and astringent taste of tea.

Theoretical Study on Storage and Release of Firefly Luciferin.

Among numerous bioluminescent organisms, firefly is the most studied one. Recent experiment proposed that sulfoluciferin (SLH2 ) may serve as a storage form of luciferin (LH2 ). In the present article, we employed density functional theory calculation to uncover the mechanism and detailed process of the storage and release reactions. Due to lack of available crystallographic structure of the related enzyme, the calculation was performed on a model system. For the storage reaction, possible amino acid residues were used for imitating the protein environment. For the release reaction, the dielectric constant of 3.0 was employed to simulate the polarity of the protein cavity. The computational results indicated that the reactions from LH2 to SLH2 and from SLH2 to LH2 are both exergonic, which favor the storage and release processes and coincide with the experimental observation. Basing on experimental and current theoretical study, we supplemented the stages of LH2 storage and release in the entire bioluminescent cycle of firefly. The current theoretical calculation could inspire the study on LH2 storage and release of other bioluminescent organisms.

Exploring the Molecular Mechanism of Astragali Radix-Curcumae Rhizoma against Gastric Intraepithelial Neoplasia by Network Pharmacology and Molecular Docking.

BACKGROUND: Astragali Radix-Curcumae Rhizoma (ARCR), a classic drug pair, has been widely used for the treatment of gastric intraepithelial neoplasia (GIN) in China. However, the underlying mechanisms of this drug pair are still unknown. Thus, elucidating the molecular mechanism of ARCR for treating GIN is imperative. METHODS: The active components and targets of ARCR were determined from the TCMSP database, and the differentially expressed genes related to GIN were identified from the GSE130823 dataset. The protein-protein interaction (PPI) network and ARCR-active component-target-pathway network were constructed by STRING 11.0 and Cytoscape 3.7.2, respectively. In addition, a receiver operating characteristic curve (ROC) was conducted to verify the key targets, and enrichment analyses were performed using R software. Molecular docking was carried out to test the binding capacity between core active components and key targets. RESULTS: 31 active components were obtained from ARCR, among which 22 were hit by the 51 targets associated with GIN. Gene Ontology (GO) functional enrichment analysis showed that biological process (BP), molecular function (MF), and cellular component (CC) were most significantly enriched in response to a drug, catecholamine binding, and apical part of the cell, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated ARCR against GIN through regulation of neuroactive ligand-receptor interaction, nitrogen metabolism, calcium signaling pathway, chemical carcinogenesis-receptor activation, drug metabolism, gap junction, and cancers. In the PPI network, 15 potential targets were identified, of which nine key targets were proven to have higher diagnostic values in ROC. Molecular docking revealed a good binding affinity of active components (quercetin, bisdemethoxycurcumin, and kaempferol) with the corresponding targets (CYP3A4, CYP1A1, HMOX1, DRD2, DPP4, ADRA2A, ADRA2C, NR1I2, and LGALS4). CONCLUSION: This study revealed the active components and molecular mechanism by which ARCR treatment is effective against GIN through regulating multipathway, such as neuroactive ligand-receptor interaction, nitrogen metabolism, and calcium signaling pathway.

CsHCT-Mediated Lignin Synthesis Pathway Involved in the Response of Tea Plants to Biotic and Abiotic Stresses.

Many studies have shown that phenolic compounds such as lignin and flavonoids enhance plant resistance. Tea plants are rich in flavonoid compounds. Whether these compounds are related to tea plant resistance is unclear. In this study, an interesting conclusion was drawn on the basis of experimental results: in response to abiotic stress (except for sucrose treatment), gene expression was increased in the phenylpropanoid and lignin pathways and was reduced in the flavonoid pathway in tea plants. CsHCTs, the genes located at the branch point of the lignin and flavonoid pathways, are most suitable for regulating the ratio of carbon flow in the lignin pathway and flavonoid synthesis. Enzymatic and genetic modification experiments proved that CsHCTs encode hydroxycinnamoyl-coenzyme A:shikimate/quinate hydroxycinnamoyl transferase in vitro and in vivo. Furthermore, the genetic modification results showed that the contents of phenolic acids and lignin were increased in tobacco and Arabidopsis plants overexpressing CsHCTs, whereas the content of flavonol glycosides was decreased. Both types of transgenic plants showed resistance to many abiotic stresses and bacterial infections. We speculate that CsHCTs participate in regulation of the metabolic flow of carbon from the flavonoid pathway to the chlorogenic acid, caffeoylshikimic acid, and lignin pathways to increase resistance to biotic and abiotic stresses.

Curcumin Alleviates Cerebral Ischemia-reperfusion Injury by Inhibiting NLRP1-dependent Neuronal Pyroptosis.

BACKGROUND: Cerebral ischemia-reperfusion injury is caused by a blood reperfusion injury in the ischemic brain and usually occurs in the treatment stage of ischemic disease, which can aggravate brain tissue injury. OBJECTIVE: Curcumin was reported to exert a good therapeutic effect on neural cells against ischemia- reperfusion injury, However, the mechanism is not clear. METHODS: In this study, Oxygen-Glucose Deprivation (OGD) model of fetal rat cerebral cortical neurons and the Middle Cerebral Artery Occlusion (MCAO) model of rats were employed to mimic cerebral ischemia-reperfusion injury in vitro and in vivo, respectively. RESULTS: We confirmed that curcumin has a promotive effect on neuronal proliferation and an inhibitory effect on neuronal pyroptosis. Furthermore, we found that curcumin could improve cerebral infarction. The results of western blotting showed that curcumin down-regulated the expression of nucleotide-binding oligomerization domain-containing protein-, leucine-rich repeats-, and pyrin domain-containing protein 1 (NLRP1), cysteinyl aspartate-specific protease 1 (caspase-1), gasdermin D (GSDMD), IL-1ß, IL-6, TNF-α, and iNOS proteins in OGD and MCAO models. NLRP1- dependent neuronal pyroptosis played an important role in cerebral ischemia-reperfusion injury. CONCLUSION: Curcumin could effectively inhibit NLRP1-dependent neuronal pyroptosis by suppressing the p38 MAPK pathway and therefore exerted neuroprotective effects against cerebral ischemia- reperfusion injury.

The herbal agent plantamajoside, exerts a potential inhibitory effect on the development of hepatocellular carcinoma.

Plantamajoside (PMS), a major component of Plantago asiatica L, has several pharmacological properties, including anti-proliferative, anti-inflammatory and anti-tumor effects. However, the effects of PMS on hepatocellular carcinoma (HCC) have yet to be determined. The aim of the present study was to investigate the effects of PMS on HCC and elucidate the underlying mechanism. All assays were conducted using 5 groups, namely control, sorafenib, and PMS 100, 50, and 25 µg/ml groups. Cell proliferation was determined by the MTT assay. Cell migration was evaluated with the wound healing and Transwell assays, respectively. Cell apoptosis and cell cycle distribution were evaluated via flow cytometry. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis and western blotting were used to further investigate the mechanism of action of PMS. Sorafenib and PMS both significantly attenuated the proliferation and migration of HCC cells, and markedly promoted cell apoptosis. PMS induced cell cycle arrest in the G0/G1 phase. The efficacy of PMS increased in a dose-dependent manner. Further study evaluated the expression of peroxisome proliferator-activated receptor (PPARγ), nuclear factor (NF)-κB and cyclooxygenase (Cox-2) using RT-qPCR analysis and western blotting. The results demonstrated that PMS promoted the expression of PPARγ and suppressed the expression of NF-κB and Cox-2. In conclusion, PMS was shown to affect cell proliferation, migration, apoptosis and cell cycle distribution. Furthermore, PMS promoted the expression of PPARγ and inhibited the expression of NF-κB and Cox-2, which may be the mechanism underlying its biological effects. Based on the results of the present study, PMS appears to be a promising agent for HCC therapy.

Randomised clinical trial: Efficacy and safety of Qing-Chang-Hua-Shi granules in a multicenter, randomized, and double-blind clinical trial of patients with moderately active ulcerative colitis.

Qing-Chang-Hua-Shi (QCHS) is a Chinese herbal formula, which is composed of 11 herbs. Studies have also shown that QCHS granules can alleviate colitis in animal models by preventing inflammatory responses and suppressing apoptosis through the MEK/ERK signaling pathway. To determine the efficacy and safety of QCHS granules in patients with moderately active UC. We performed a multicenter, randomized, placebo-controlled, double-blind study of patients with moderately active UC who did not respond to 4 weeks of mesalazine therapy at the maximum dose. Patients were randomly assigned to groups and administered QCHS granules (125 g/day, n = 59) or an identical placebo, which was similar to the QCHS granules in color and taste (125 g/day, n = 60), with continued 5-ASA 4 g/d therapy for 12 weeks. The primary outcome was the rate of clinical response and clinical remission at week 12. The secondary outcomes were health-related quality of life, endoscopic response rate, and mucosal healing rate. Any changes in mucus/bloody stool and diarrhea were recorded. Out of the 119 enrolled patients at 10 different centers in China, 102 patients completed the trial. Clinical remission and clinical response were seen in 31.48% and 92.59% of QCHS-treated patients, and 12.50% and 72.92% of placebo-treated patients, respectively. There was a significant difference between the two treatment groups. More patients receiving QCHS granules vs. placebo achieved remission of mucus/bloody stool (70.37% vs. 47.92%, P = 0.0361). Adverse event rates were similar (QCHS granules 38.33%; placebo 25.42%). In conclusion, QCHS granules were superior to the placebo in introducing clinical remission and mucosal healing, as well as in relieving mucus/blood stool in patients with moderately active and 5-ASA-refractory UC.

Functional characterization of three flavonol synthase genes from Camellia sinensis: Roles in flavonol accumulation.

Flavonol derivatives are a group of flavonoids benefiting human health. Their abundant presence in tea is associated with astringent taste. To date, mechanism pertaining to the biosynthesis of flavonols in tea plants remains unknown. In this study, we used bioinformatic analysis mining the tea genome and obtained three cDNAs that were annotated to encode flavonol synthases (FLS). Three cDNAs, namely CsFLSa, b, and c, were heterogenously expressed in E. coli to induce recombinant proteins, which were further used to incubate with three substrates, dihydrokampferol (DHK), dihydroquercetin (DHQ), and dihydromyricetin (DHM). The resulting data showed that three rCsFLSs preferred to catalyze (DHK). Overexpression of each cDNA in tobacco led to the increase of kampferol and the reduction of anthocyanins in flowers. Further metabolic profiling of flavan-3-ols in young tea shoots characterized that kaempferol derivatives were the most abundant, followed by quercetin and then myricetin derivatives. Taken together, these data characterized the key step committed to the biosynthesis of flavonols in tea leaves. Moreover, these data enhance understanding the metabolic accumulation relevance between flavonols and other main flavonoids such as flavan-3-ols in tea leaves.

Proanthocyanidin-Aluminum Complexes Improve Aluminum Resistance and Detoxification of Camellia sinensis.

Aluminum (Al) influences crop yield in acidic soil. The tea plant (Camellia sinensis) has high Al tolerance with abundant monomeric catechins in its leaves, especially epigallocatechin gallate (EGCG), and polymeric proanthocyanidins in its roots (rPA). The role of these polyphenols in the Al resistance of tea plants is unclear. In this study, we observed that these polyphenols could form complexes with Al in vitro, and complexation capacity was positively influenced by high solution pH (pH 5.8), polyphenol type (rPA and EGCG), and high Al concentration. In the 27Al nuclear magnetic resonance (NMR) experiment, rPA-Al and EGCG-Al complex signals could be detected both in vitro and in vivo. The rPA-Al and EGCG-Al complexes were detected in roots and old leaves, respectively, of both greenhouse seedlings and tea garden plants. Furthermore, in seedlings, Al accumulated in roots and old leaves and mostly existed in the apoplast in binding form. These results indicate that the formation of complexes with tea polyphenols in vivo plays a vital role in Al resistance in the tea plant.

Comparative analysis of phenolic compound metabolism among tea plants in the section Thea of the genus Camellia.

Wild tea plants, which are classified into different species in the section Thea of the genus Camellia, are widely distributed in southern China. Tea produced from these plants has a unique flavor, which is different from that of tea produced from tea cultivars. In this study, we performed a comparative analysis of morphology, phylogenetic relationships, and phenolic compound metabolism between two wild tea plants (Gujing and Siqiu) and a tea cultivar (Shuchazao). Siqiu and Gujing tea plants had similar morphological traits and could be phylogenetically classified into a same cluster, which was entirely separate from the cluster containing widely cultivated cultivars such as Camellia sinensis cv. Shuchazao. Combined metabolomic and transcriptome analyses revealed that UGT84a22 was highly expressed in Gujing leaves compared with Shuchazao and Siqiu leaves, which may lead to the high accumulation of galloylquinic acid in Gujing leaves. A 14-bp deletion spanning the -765-(-7 5 1) range in the F3'5'H promoter potentially led to low F3'5'H expression levels in Siqiu and Gujing tea plants, which severely disrupted the accumulation of trihydroxy flavonoids in Gujing and Siqiu tea leaves. The high astringency intensity in Gujing tea could be due to the high accumulation of proanthocyanidins and galloylquinic acid. The results of the present study may improve our understanding of the metabolic characteristics of each evolutionary group of species or varieties in the section Thea of the genus Camellia.