Widely Untargeted Metabolomics Profiling Combined with Transcriptome Analysis Provides New Insight into Amino Acid Biosynthesis at Different Developmental Stages of Rubus Chingii Hu (Chinese Raspberry).

The composition and profile of amino acids in Rubus chingii (R. chingii) Hu serve as critical indicators of its nutritional quality. A comprehensive understanding of the amino acid metabolism within R. chingii is instrumental in the formulation and innovation of functional foods derived from this species. Utilizing advanced techniques such as wide-ranging untargeted metabolomics, transcriptome analysis, interaction network mapping, heat map analysis, and quantitative real-time PCR, we conducted a comprehensive assessment of the quality attributes across four distinct developmental stages of R. chingii. Our meticulous analysis uncovered a rich tapestry of 76 distinct amino acids and their derivatives within the developmental stages of R. chingii. The spectrum of essential amino acids was not only broad but also displayed a high degree of variety. Notably, leucine, lysine, and phenylalanine stood out as the most abundant amino acids, underscoring their significant presence throughout the growth cycle of R. chingii. The proportion of essential amino acids relative to the total amino acid content in R. chingii exhibited a notable trajectory of change throughout its developmental stages. It began with 30.92% in the immature green phase, advanced to 31.04% during the transition from green to yellow, peaked at 33.62% in the yellow to red stage, and then moderated to 30.43% in the full red phase. This pattern suggests a strategic modulation of amino acid composition, aligning with the evolving nutritional requirements and metabolic shifts as the fruit matures. Concurrent analysis of interaction networks and heat maps, alongside comprehensive profiling of amino acid metabolism and transcriptomic examination, was conducted to elucidate the intricate dynamics of cellular processes. The results showed that seven differentially expressed genes (DEGs) played important roles in amino acid metabolism, including PFK, BCAT1, TSB, ASA, ACO, TOM2AH3, and BCAT2. The expression patterns of seven DEGs conformed closely to the findings revealed by the preceding RNA-seq analysis. In this investigation, we elucidated the regulatory mechanisms underlying amino acid metabolism across the four distinct developmental stages of R. chingii through comprehensive amino acid profiling and transcriptomic analysis. These insights lay the groundwork for the development of novel functional food applications utilizing R. chingii.

Contrastive representation learning on dynamic networks.

Representation learning for dynamic networks is designed to learn the low-dimensional embeddings of nodes that can well preserve the snapshot structure, properties and temporal evolution of dynamic networks. However, current dynamic network representation learning methods tend to focus on estimating or generating observed snapshot structures, paying excessive attention to network details, and disregarding distinctions between snapshots with larger time intervals, resulting in less robustness for sparse or noisy networks. To alleviate these challenges, this paper proposes a contrastive mechanism for temporal representation learning on dynamic networks, inspired by the success of contrastive learning in visual and static network representation learning. This paper proposes a novel Dynamic Network Contrastive representation Learning (DNCL) model. Specifically, contrast objective functions are constructed using intra-snapshot and inter-snapshot contrasts to capture the network topology, node feature information, and network evolution information, respectively. Rather than estimating or generating ground-truth network features, the proposed approach maximizes mutual information between nodes from different time steps and views generated. The experimental results of link prediction, node classification, and clustering on several real-world and synthetic networks demonstrate the superiority of DNCL over state-of-the-art methods, indicating the effectiveness of the proposed approach for dynamic network representation learning.

Bioinformatic Analysis of Key Regulatory Genes in Adult Asthma and Prediction of Potential Drug Candidates.

Asthma is a common chronic disease that is characterized by respiratory symptoms including cough, wheeze, shortness of breath, and chest tightness. The underlying mechanisms of this disease are not fully elucidated, so more research is needed to identify better therapeutic compounds and biomarkers to improve disease outcomes. In this present study, we used bioinformatics to analyze the gene expression of adult asthma in publicly available microarray datasets to identify putative therapeutic molecules for this disease. We first compared gene expression in healthy volunteers and adult asthma patients to obtain differentially expressed genes (DEGs) for further analysis. A final gene expression signature of 49 genes, including 34 upregulated and 15 downregulated genes, was obtained. Protein-protein interaction and hub analyses showed that 10 genes, including POSTN, CPA3, CCL26, SERPINB2, CLCA1, TPSAB1, TPSB2, MUC5B, BPIFA1, and CST1, may be hub genes. Then, the L1000CDS2 search engine was used for drug repurposing studies. The top approved drug candidate predicted to reverse the asthma gene signature was lovastatin. Clustergram results showed that lovastatin may perturb MUC5B expression. Moreover, molecular docking, molecular dynamics simulation, and computational alanine scanning results supported the notion that lovastatin may interact with MUC5B via key residues such as Thr80, Thr91, Leu93, and Gln105. In summary, by analyzing gene expression signatures, hub genes, and therapeutic perturbation, we show that lovastatin is an approved drug candidate that may have potential for treating adult asthma.

Study on enhancement of hemoglobin antitoxic ability modified with chromium and ruthenium.

Hemoglobin is essential for carrying oxygen (O2) in the blood. However, its ability to bind excessively to carbon monoxide (CO) makes it susceptible to CO poisoning. To reduce the risk of CO poisoning, Cr-based heme and Ru-based heme were selected from among many transition metal-based hemes based on their characteristics of adsorption conformation, binding intensity, spin multiplicity, and electronic properties. The results showed that hemoglobin modified by Cr-based heme and Ru-based heme had strong anti-CO poisoning abilities. The Cr-based heme and Ru-based heme exhibited much stronger affinity for O2 (-190.67 kJ/mol and -143.18 kJ/mol, respectively) than Fe-based heme (-44.60 kJ/mol). Moreover, Cr-based heme and Ru-based heme exhibited much weaker affinity for CO (-121.50 kJ/mol and -120.88 kJ/mol, respectively) than their affinity for O2, suggesting that they were less likely to cause CO poisoning. The electronic structure analysis also supported this conclusion. Additionally, molecular dynamics analysis showed that hemoglobin modified by Cr-based heme and Ru-based heme was stable. Our findings offer a novel and effective strategy for enhancing the reconstructed hemoglobin's ability to bind O2 and reduce its potential for CO poisoning.

Photoacoustic/ultrasonic dual-mode imaging for monitoring angiogenesis and synovial erosion in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by the formation of new vessels, synovial proliferation and destruction of articular cartilage. However, characteristic early diagnostic and therapeutic monitoring methods are still lacking. We report a study using a photoacoustic/ultrasound (PA/US) dual-mode imaging for RA disease. By establishing a collagen-induced (CIA) RA mouse model to classify disease states based on a subjective grading system, PA/US imaging allows real-time assessment of synovial erosion and vascular opacification within the knee joint in different disease states at high spatial resolution. The system also quantitatively monitors subcutaneous vascular physiology and morphology in the hind paw of mice, measuring the area and photoacoustic signal intensity of vascular proliferation and showing a positive correlation with disease grading. Compared to traditional subjective scoring of arthritis severity, the PA/US imaging is more sensitive i.e., vascular signals and synovial erosion can be observed early in the course of arthritis.

Impact of dyslipidemia on the severity of symptomatic lumbar spine degeneration: A retrospective clinical study.

Background: Lumbar intervertebral disc degeneration (IVDD) is an important cause of low back pain or sciatica, and metabolic factors play an important role. However, little is known about the relationship of dyslipidemia to the risk of intervertebral disc degeneration (IVDD). This study aimed to assess the impact of serum lipid levels on the severity of lumbar disc degeneration and to investigate its association with endplate inflammation. Methods: We conducted a case retrospective study in which a total of 302 hospitalized Chinese patients were recruited, of whom 188 (112 males and 76 females; mean age: 51.66 years) were without underlying disease, while the remaining 114 patients (51 males and 63 females; mean age: 62.75 years) had underlying diseases. We examined fasting serum lipid levels for total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). Magnetic resonance imaging (MRI) was used to determine endplate inflammation. Pfirrmann grading and Weishaupt grading were used to evaluate the severity of intervertebral disc degeneration and facet joint degeneration, respectively. Results: There was no difference in age, gender, and general BMI between the two groups (P > 0.05), but there were significantly high levels in TC, LDL-C, and LDL-C/HDL-C (P = 0.04, P = 0.013, P = 0.01, respectively). TG and HDL-C showed no significant difference (P = 0.064, P = 0.336, respectively). The multivariate logistic regression model showed that age was a risk factor for the occurrence of endplate inflammation. In the group without underlying diseases, age, but not other indicators, was a risk factor for the occurrence of endplate inflammation (P < 0.01), In the group with underlying diseases, none of the patient indicators was directly related to the occurrence of endplate inflammation (P > 0.05). A nonlinear machine learning model was used to measure the contribution of each factor to the disease outcome and to analyze the effect between the top three contributing factors and the outcome variables. In patients without underlying diseases, the top three factors contributing to the severity grading of intervertebral disc degeneration were age (32.9%), high-density lipoproteins (20.7%), and triglycerides (11.8%). For the severity grading of facet joint degeneration, the top three contributing factors were age (27.7%), high-density lipoproteins (19.4%), and triglycerides (14.6%). For patients with underlying diseases, the top three factors contributing to intervertebral disc degeneration were age (25.4%), BMI (15.3%), and low-density lipoprotein/high-density lipoprotein ratio (13.9%). In terms of degree classification for facet joint degeneration, the top three contributing factors were age (17.5%), BMI (17.2%), and total cholesterol (16.7%). Conclusion: This study shows that age, high-density lipoprotein, and triglycerides affect the degree of degeneration in patients with symptomatic lumbar degeneration without underlying diseases. Age and BMI are two major factors affecting the severity of degeneration in patients with underlying diseases, and dyslipidemia is a secondary factor. However, there is no clear association between dyslipidemia and the occurrence of endplate inflammation in either group.

Natural Deep Eutectic Solvent-Based Microwave-Assisted Extraction of Total Flavonoid Compounds from Spent Sweet Potato (Ipomoea batatas L.) Leaves: Optimization and Antioxidant and Bacteriostatic Activity.

Natural deep eutectic solvents (NADESs) coupled with microwave-assisted extraction (MAE) were applied to extract total flavonoid compounds from spent sweet potato (Ipomoea batatas L.) leaves. In this study, ten different NADESs were successfully synthesized for the MAE. Based on single-factor experiments, the response surface methodology (RSM) was applied, and the microwave power, extraction temperature, extraction time, and solid−liquid ratio were further evaluated in order to optimize the yields of total flavonoid compounds. Besides, the extracts were recovered by macroporous resin for the biological activity detection of flavonoid compounds. As a result, NADES-2, synthesized by choline chloride and malic acid (molar ratio 1:2), exhibited the highest extraction yield. After that, the NADES-2-based MAE process was optimized and the optimal conditions were as follows: microwave power of 470 W, extraction temperature of 54 °C, extraction time of 21 min, and solid−liquid ratio of 70 mg/mL. The extraction yield (40.21 ± 0.23 mg rutin equivalents/g sweet potato leaves) of the model validation experiment was demonstrated to be in accordance with the predicted value (40.49 mg rutin equivalents/g sweet potato leaves). In addition, flavonoid compounds were efficiently recovered from NADES-extracts with a high recovery yield (>85%) using AB-8 macroporous resin. The bioactivity experiments in vitro confirmed that total flavonoid compounds had good DPPH and O2−· radical-scavenging activity, as well as inhibitory effects on E. coli, S. aureus, E. carotovora, and B. subtilis. In conclusion, this study provides a green and efficient method to extract flavonoid compounds from spent sweet potato leaves, providing technical support for the development and utilization of sweet potato leaves' waste.

Inhibitory Effect of Lactobacillus delbrueckii subsp. bulgaricus KSFY07 on Kappa-Carrageenan-Induced Thrombosis in Mice and the Regulation of Oxidative Damage.

A mouse thrombosis model was established by kappa-carrageenan to observe the inhibitory effect of Lactobacillus delbrueckii subsp. bulgaricus KSFY07 (LDSB-KSFY07) on thrombosis and the oxidative stress response. Mouse serum, liver tissue-related indicators, and intestinal microbial composition were measured by examining the expression of microbes in mouse faeces using a biochemical kit, slice observations, and quantitative polymerase chain reaction (qPCR) experiments. The results showed that LDSB-KSFY07 effectively reduced the degree of black tail in thrombotic mice, increased activated partial thromboplastin time (APTT), and decreased thrombin time (TT), fibrinogen (FIB), and prothrombin time (PT) in thrombotic mice. LDSB-KSFY07 was also able to reduce malondialdehyde (MDA) levels and increase superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) levels in the serum and liver tissues of thrombotic mice. Pathological observations showed that LDSB-KSFY07 reduced liver tissue lesions and tail vein thrombosis. Further, experimental results showed that LDSB-KSFY07 was able to upregulate the mRNA expression of copper/zinc-SOD (Cu/Zn-SOD), manganese-SOD, and GSH-Px in the liver tissue of thrombotic mice. Moreover, LDSB-KSFY07 was also able to downregulate the mRNA expression of NF-κB p65, intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin in tail vein vascular tissue. Meanwhile, LDSB-KSFY07 could raise plasminogen activator inhibitor-1 (PAI-1) mRNA expression and reduce tissue plasminogen activator (t-PA) expression in heart and tail vein vascular tissues of thrombotic mice. A mouse faeces examination revealed that LDSB-KSFY07 could also upregulate Bacteroides, Lactobacterium, and Bifidobacterium microbial expression and downregulate Firmicutes expression in the gut. These results indicate that LDSB-KSFY07 was able to inhibit mouse thrombosis and reduce liver oxidative stress damage in thrombus mice and show that high concentrations of LDSB-KSFY07 provided a better response similar to that of the drug heparin.

Effect of Lactobacillus fermentum HFY06 Combined with Arabinoxylan on Reducing Lipid Accumulation in Mice Fed with High-Fat Diet.

In this experiment, a high-fat diet was used to induce hyperlipidemia in mice to determine the synergistic effect of AX and L. fermentum HFY06 on the prevention of hyperlipidemia and its potential regulatory mechanism. The results of this study showed that after the AX and L. fermentum HFY06 synergistic intervention, the body weight, epididymal fat index, blood lipid level, and liver function indexes of mice were improved. In addition, the synbiotics comprising AX and L. fermentum HFY06 increased the CAT activity in the serum of mice on a high-fat diet, reduced NO and MDA levels, and improved the body's oxidative stress. From the perspective of molecular biology, on the one hand, AX and L. fermentum HFY06 synergistic intervention activated the AMPK pathway to regulate body lipid metabolism; up-regulated the mRNA expressions of CPT-1, PPAR-α, CYP7A1, and HSL; and down-regulated the mRNA expressions of ACC, C/EBPα, and LPL. On the other hand, the synergistic effect of AX and HFY06 enhanced the mRNA expressions of ZO-1, occludin, and claudin-1 in the small intestine of mice, increased the strength of the intestinal barrier, and optimized the composition of the intestinal microbiota. From the above results, it can be concluded that AX and L. fermentum HFY06 have a synergistic effect in improving hyperlipidemia. However, this study was only performed using animal models, and the lipid synthesis and metabolism mechanism are complicated; hence, further clinical studies are needed.

Quality evaluation of Tetrastigma hemsleyanum different parts based on quantitative analysis of 42 bioactive constituents combined with multivariate statistical analysis.

INTRODUCTION: The root of Tetrastigma hemsleyanum (RTH) has been widely used as a folk medicine in China. Meanwhile, its stems (STH) and leaves (LTH) are consumed as functional tea and food supplementation. Therefore, it is important to get a better understanding of the distribution of bioactive constituents in different parts of T. hemsleyanum. OBJECTIVE: To develop a method for quantitative analysis of multiple bioactive constituents and comparing their distribution in RTH, STH and LTH. METHODS: Ultra-performance liquid chromatography triple quadrupole ion trap tandem mass spectrometry (UPLC-QTRAP-MS/MS) was used for the quantitative analysis. The quantitative data were further analysed by principal component analysis (PCA), hierarchical cluster analysis (HCA) and partial least squares determinant analysis (PLS-DA). RESULTS: Forty-two constituents in RTH, STH and LTH, including 14 flavonoids, three phenolic acids, 15 amino acids and 10 nucleosides, were quantitatively determined. The contents of flavonoids and phenolic acids in LTH were significantly higher than those in RTH and STH. While the contents of amino acids and nucleosides in LTH were less than those in RTH and STH. Multivariate statistical analysis can significantly classify and distinguish RTH, STH, and LTH. CONCLUSIONS: The present method would be helpful for the quality control of T. hemsleyanum, and the results would be useful for the efficient utilisation of T. hemsleyanum in the future.

Discovery and identification of potential anti-melanogenic active constituents of Bletilla striata by zebrafish model and molecular docking.

BACKGROUND: Bletilla striata is the main medicine of many skin whitening classic formulas in traditional Chinese medicine (TCM) and is widely used in cosmetic industry recently. However, its active ingredients are still unclear and its fibrous roots are not used effectively. The aim of the present study is to discover and identify its potential anti-melanogenic active constituents by zebrafish model and molecular docking. METHODS: The antioxidant activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2'-azino-bis-(3-ethylbenthiazoline-6-sulphonic acid) (ABTS) radical scavenging activity and ferric reducing antioxidant power (FRAP) assay. The anti-melanogenic activity was assessed by tyrosinase inhibitory activity in vitro and melanin inhibitory in zebrafish. The chemical profiles were performed by ultra-high-performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Meanwhile, the potential anti-melanogenic active constituents were temporary identified by molecular docking. RESULTS: The 95% ethanol extract of B. striata fibrous roots (EFB) possessed the strongest DPPH, ABTS, FRAP and tyrosinase inhibitory activities, with IC50 5.94 mg/L, 11.69 mg/L, 6.92 mmol FeSO4/g, and 58.92 mg/L, respectively. In addition, EFB and 95% ethanol extract of B. striata tuber (ETB) significantly reduced the melanin synthesis of zebrafish embryos in a dose-dependent manner. 39 chemical compositions, including 24 stilbenoids were tentatively identified from EFB and ETB. Molecular docking indicated that there were 83 (including 60 stilbenoids) and 85 (including 70 stilbenoids) compounds exhibited stronger binding affinities toward tyrosinase and adenylate cyclase. CONCLUSION: The present findings supported the rationale for the use of EFB and ETB as natural skin-whitening agents in pharmaceutical and cosmetic industries.

Comparative pharmacokinetic study of six lignans in normal and diabetic rats after oral administration of Saururus chinensis extract by LC-MS/MS.

Saururus chinensis (SC) possesses significant anti-diabetic activity and lignans were its major bioactive compounds. In this study, a rapid and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for simultaneous quantification of six lignans, namely (-)-(7R,8R)-machilin D (1), verrucesin (2), rel-(7S,8S,7'R,8'R)-3,3',4,4',5,5'-hexamethoxy-7.O.7',8.8'-lignan (3), manassantin A (4), manassantin B (5), and saucerneol F (6) in rat's plasma. It was validated with acceptable linearity (r ≥ 0.9922), accuracy (80.42-95.17%), precision (RSD ≤ 12.08%), and extraction recovery (80.36-93.45%). The method was successfully applied to the comparative pharmacokinetic study of the six lignans in normal and diabetic rats after oral administration of SC extract. Results showed that the areas under the plasma concentration-time curve (AUC0 → t and AUC0 → ∞ ) of (-)-(7R,8R)-machilin D, rel-(7S,8S,7'R,8'R)-3,3',4,4',5,5'-hexamethoxy-7.O.7',8.8'-lignan, manassantin B, and saucerneol F in diabetic rats were significantly increased, and the plasma clearance (CL) of (-)-(7R,8R)-machilin D in diabetic rats was significantly decreased. However, the AUC0 → t and AUC0 → ∞ of verrucesin were significantly decreased, and its CL was significantly increased in diabetic rats compared with those in normal rats. These results indicated that there were remarkable differences in the pharmacokinetic parameters between the normal and diabetic rats. The pharmacokinetic studies might be beneficial for the clinical use of SC as hypoglycemic agent.

Acute and subacute toxicity evaluation of Houttuynia cordata ethanol extract and plasma metabolic profiling analysis in both male and female rats.

Houttuynia cordata has been used as a traditional medicine for more than 1500 years. It has aroused wide public concern about its safety in the past few years, for it contains various aristolactams. However, the safety of H. cordata extract remains unclear. In the present study, single dose (2000 mg/kg) and subacute (250, 500, and 1000 mg/kg/day for 28 days) oral toxicity studies of the 95% ethanol extract of H. cordata (HCE) were performed in both male and female Sprague-Dawley (SD) rats. Hematological, biochemical, histopathological parameters, and plasma metabolic profiling were assessed. The single-dose toxicity of HCE was more than 2000 mg/kg. The subacute toxicity results showed that no significant adverse effect of HCE was observed at 250 mg/kg/day. However, five rats died in 500 and 1000 mg/kg/day groups and exhibited toxicities to liver and kidney. Plasma metabolic profiling analysis suggested that a number of metabolic disturbances were induced by oral administration of HCE, focusing on energy metabolism, amino acid metabolism, and lipids metabolism. Moreover, it appeared that male rats were more susceptible to the toxic effects of HCE than female rats. Therefore, in this preliminary study, oral administration of HCE 250 mg/kg/day can be regarded as the no observed adverse effect level in rats over 28 days. However, long-term use of HCE with large doses exhibited some hepatotoxicity and nephrotoxicity in rats.

Stromal cell-derived factor-1 may play pivotal role in distraction-stimulated neovascularization of diabetic foot ulcer.

Diabetic foot ulcer (DFU) has become a major medical, social and economic concern worldwide. It is highly desirable to develop promising new solutions to effectively and appropriately treat DFU. In recent years, investigators have used an innovative technology called proximal tibial cortex transverse distraction (PTCTD) to treat DFU and have achieved satisfactory results in terms of improved wound healing and circumvention of amputation as a consequence of enhanced neovascularization and perfusion of the ulcerated feet after the operation, but the underlying mechanism has not been explored. Previous studies have suggested that in addition to stimulating osteogenesis, bone distraction also facilitates neovascularization, which may be associated with the chemokine stromal cell-derived factor-1 (SDF-1). As an important member of the chemokine family, SDF-1 is primarily responsible for the homing and migration of endothelial progenitor cells (EPCs) or bone marrow-derived mesenchymal stem cells (BMSCs), and plays a central role in the process of neovascularization. In vivo or in vitro experiments show that bone distraction can induce the expression of SDF-1 and increase its plasma concentration. Moreover, some researchers have found that an insufficient level of SDF-1 in the circulation and wounds of patients with DFU can lead to impaired neovascularization. Therefore, we believe that SDF-1 plays an important role in promoting neovascularization of DFU as a result of bone distraction. We summarize the currently relevant literature to put forward an undisclosed but meaningful mechanism of bone distraction in the treatment of DFU.

Further understanding on osteopetrotic femoral fractures: a case report and literature review.

BACKGROUND: Osteopetrosis is a genetic disease characterized by defects in osteoclast formation and function. There were a few cases of subtrochanteric femur fractures treated with dynamic hip screw (DHS) in patients with osteopetrosis, but unfortunately the healing outcome was rather poor. CASE PRESENTATION: We present our experience for treating a patient with intermediate autosomal recessive osteopetrosis (IRO) suffering from subtrochanteric femur fracture. In this case, we successfully used dynamic hip screw (DHS) internal fixation through meticulous preoperative planning and postoperative care, as well as application of surgical techniques. The patient displayed stable internal fixation with no limitation of activities during follow-up for 15 months. In addition to this case, a review of previous case reports showed an increasing number of case reports demonstrating that surgical treatment-related complications could be avoided preoperatively, intraoperatively, and postoperatively. CONCLUSION: DHS for this patient, who suffered from subtrochanteric fractures with osteopetrosis, was successfully implemented. In the light of a comprehensive literature review, preoperative planning, surgical techniques, and postoperative rehabilitation care can significantly reduce the complications.

Simultaneous determination of multiple active 2-(2-phenylethyl)chromone analogues in agarwood by HPLC, QAMS, and UPLC-MS.

INTRODUCTION: Chromones are the major constituents of agarwood and are considered to be directly related to its quality. Agarotetrol, a chromone derivative, is a Chinese Pharmacopoeia content detection index. However, comprehensive high-performance liquid chromatography (HPLC), quantitative analysis of multiple components by a single marker (QAMS), and ultra-performance liquid chromatography mass spectrometry (UPLC-MS) analyses of this pharmacopeial plant material have never been performed. Moreover, reports regarding the separation and detection of multiple active 2-(2-phenylethyl)chromone analogues from this plant material are surprisingly scarce. OBJECTIVE: To establish a simple, reliable, and effective HPLC method utilising both diode array and MS detection for the simultaneous determination of multiple active chromone analogues in agarwood. METHODS: Four 2-(2-phenylethyl)chromones were isolated from methanol extracts of agarwood. After optimising the extraction, separation, and analytical conditions, validation of the developed analytical method indicated good linearity, satisfactory precision, and good recovery. On this basis, a method for the quantitative analysis of multiple components by a single marker was established. The four 2-(2-phenylethyl)chromones were identified by nuclear magnetic resonance spectroscopic analysis and UPLC coupled to electrospray ionisation quadrupole-time-of-flight MS. CONCLUSIONS: The behaviour of the chromones characterised by MS fragmentation indicated a loss of molecular CO and the formation of m/z 121 compounds by the cleavage of CH2 -CH2 bonds between the chromone and phenyl moieties. Three detection methods were successfully used in this study for agarwood detection, and this protocol may potentially be used as a tool for the quality control of agarwood.

[The efficacy of hyperbaric oxygen in the treatment of sudden sensorineural hearing loss].

Objective:To investigate the therapeutic effect of hyperbaric oxygen on sudden sensorineural hearing loss（SSNHL）. Method:One hundred and seven patients with SSNHL were selected, and were randomly divided into 3 groups according to admission order. Therapeutic schedule in group A （38 cases） consisted of hyperbaric oxygen treatment once a day for 10 consecutive days, then 1 day off treatment, and on treatment for another 10 consecutive days （20 treatments in total）. Twenty-seven patients in group B were treated with hyperbaric oxygen twice per day for a total of 10 days. Forty-two patients in group C didn't receive hyperbaric oxygen therapy. Meanwhile, all patients were given conventional treatment for 14 days in accordance with the recommendations of the otolaryngology, head and neck branch of the Chinese medical association for the diagnosis and treatment of sudden sensorineural hearing loss. Patients in group A underwent pure tone audiometry after the end of treatment, and patients in group B and C were reexamined 20 days after the start of treatment, and the treatment efficacy of the three groups was compared. Result:The overall effective rate of group A was 78.9%, and the improvement of hearing threshold was（27.2±17.5） dB HL. In group B, the overall effective rate was 81.5%, and the improvement of threshold value was（26.9±16.7） dB HL. The overall effective rate of group C was 61.9%, and the improvement of threshold value o was（17.6±18.5） dB HL. The effective rate of group C was significantly higher than that of group A or B, while there was no significant difference in efficacy between group A and group B. Conclusion:The efficacy of hyperbaric oxygen combined with drug therapy for SSNHL is better than that of drug therapy alone. Programs of different hyperbaric oxygen combined with drug therapy can be selected on an individual basis.