Sleep deprivation-induced anxiety-like behaviors are associated with alterations in the gut microbiota and metabolites.

The present study aimed to characterize the gut microbiota and serum metabolome changes associated with sleep deprivation (SD) as well as to explore the potential benefits of multi-probiotic supplementation in alleviating SD-related mental health disorders. Rats were subjected to 7 days of SD, followed by 14 days of multi-probiotics or saline administration. Open-field tests were conducted at baseline, end of SD (day 7), and after 14 days of saline or multi-probiotic gavage (day 21). Metagenomic sequencing was conducted on fecal samples, and serum metabolites were measured by untargeted liquid chromatography tandem-mass spectrometry. At day 7, anxiety-like behaviors, including significant decreases in total movement distance (P = 0.0002) and staying time in the central zone (P = 0.021), were observed. In addition, increased levels of lipopolysaccharide (LPS; P = 0.028) and decreased levels of uridine (P = 0.018) and tryptophan (P = 0.01) were detected in rats after 7 days of SD. After SD, the richness of the gut bacterial community increased, and the levels of Akkermansia muciniphila, Muribaculum intestinale, and Bacteroides caecimuris decreased. The changes in the host metabolism and gut microbiota composition were strongly associated with the anxiety-like behaviors caused by SD. In addition, multi-probiotic supplementation for 14 days modestly improved the anxiety-like behaviors in SD rats but significantly reduced the serum level of LPS (P = 0.045). In conclusion, SD induces changes in the gut microbiota and serum metabolites, which may contribute to the development of chronic inflammatory responses and affect the gut-brain axis, causing anxiety-like behaviors. Probiotic supplementation significantly reduces serum LPS, which may alleviate the influence of chronic inflammation. IMPORTANCE: The disturbance in the gut microbiome and serum metabolome induced by SD may be involved in anxiety-like behaviors. Probiotic supplementation decreases serum levels of LPS, but this reduction may be insufficient for alleviating SD-induced anxiety-like behaviors.

Somatisation Disorder Detection via Speech: Introducing a Self-Supervised Learning Model.

With the depressive psychiatric disorders becoming more common, people are gradually starting to take it seriously. Somatisation disorders, as a general mental disorder, are rarely accurately identified in clinical diagnosis for its specific nature. In the previous work, speech recognition technology has been successfully applied to the task of identifying somatisation disorders on the Shenzhen Somatisation Speech Corpus. Nevertheless, there is still a scarcity of labels for somatisation disorder speech database. The current mainstream approaches in the speech recognition heavily rely on the well labelled data. Compared to supervised learning, self-supervised learning is able to achieve the same or even better recognition results while reducing the reliance on labelled samples. Moreover, self-supervised learning can generate general representations without the need for human hand-crafted features depending on the different recognition tasks. To this end, we apply self-supervised learning pre-trained models to solve few-labelled somatisation disorder speech recognition. In this study, we compare and analyse the results of three self-supervised learning models (contrastive predictive coding, wav2vec and wav2vec 2.0). The best result of wav2vec 2.0 model achieves 77.0 % unweighted average recall and is significantly better than CPC (p < .005), performing better than the benchmark of the supervised learning model.Clinical relevance- This work proposed a self-supervised learning model to resolve the few-labelled SD speech data, which can be well used for helping psychiatrists with clinical assistant to diagnosis. With this model, psychiatrists no longer need to spend a lot of time labelling SD speech data.

Chromogenic Visualization of Single RNA Molecules In Situ with Duplex Capability by Rolling Circle Amplification with Alkaline Phosphatase.

Visualizing spatial patterns of gene expression by optical microscopy at single-molecule resolution represents a long-standing challenge for imaging and molecular engineering technologies. In this study, we developed a method for visualizing mRNA with duplex capability by optical microscopy using rolling circle amplification with streptavidin-modified alkaline phosphatase (SA-ALP) to provide highly selective and sensitive RNA detection. ALP-based RNA detection provides comparable sensitivity and specificity to fluorescence-based in situ assays and similar performance to the current RNAscope technique for single-molecule RNA detection, but with improved ease of operation. This versatile and relatively user-friendly method of single-molecule RNA visualization can also overcome common problems of background interference. Our findings show that the red spots generated by the Fast Red staining in situ are readily quantified by image analysis, even in samples with heavy melanin deposition, supporting the clinical translation of this assay to improve diagnostic assays for recalcitrant tissues. This system was adaptable for duplex assays with multiple probes and multiple stains, which is ALP with horseradish peroxidase to produce red and brown signals to simultaneously visualize two different RNA targets. The duplex assay can be successfully applied to quantify mRNA expression from two genes in situ within single cells and multiple cell types. With the advantages of high sensitivity and low hardware requirements, this versatile and user-friendly method of RNA visualization may enable low-resource institutions to conduct previously inaccessible diagnostic or research questions about the in situ expression and distribution of RNAs at single-molecule resolution.

Protocatechuic Acid Ameliorates High Fat Diet-Induced Obesity and Insulin Resistance in Mice.

SCOPE: Insulin resistance is a common feature of obesity and type 2 diabetes and partly results from an imbalance between food intake and energy expenditure. Therefore, efficient and safe insulin resistance treatment therapies are warranted. This work is aim to access the impact of protocatechuic acid (PCA), a catechol-type O-diphenol phenolic acid, in high fat diet (HFD)-induced glucose, and lipid dysregulation. METHODS AND RESULTS: Five-week-old male C57BL/6 mice are fed with HFD for 4 weeks and then are randomly divided into two cohorts: one cohort feed with HFD is free access to sterile water for 4 weeks, another cohort is free access to PCA-containing water (2.7 mM) for 4 weeks with HFD. In this study, using a hyperinsulinemic-euglycemic mouse clamp, it is showed that PCA-treated mice display improved systemic insulin resistance via enhanced fatty acid mobilization and utilization, thereby reducing ectopic lipid accumulation and promoting hepatic and peripheral insulin action. CONCLUSIONS: This study provides insights on the potent pharmacological effects of PCA from food sources on improving high fat diet (HFD)-induced whole-body insulin resistance and type 2 diabetes.

LY6E protein facilitates adeno-associated virus crossing in a biomimetic chip model of the human blood-brain barrier.

The blood-brain barrier (BBB) controls chemical access to the brain and maintains fluid homeostasis, but in vitro models accurately simulating the physiological characteristics of the BBB are lacking. Here, we describe a simple and reproducible biomimetic chip-based model of the human BBB. In this bilayer co-culture, astrocytes and brain microvascular endothelial cells (BMECs) are respectively seeded in upper and lower chambers separated by a semi-permeable membrane, with fluid shear force provided by a precision tilt shaker. Evaluation of barrier crossing by fluorescein sodium, 40 kDa or 70 kDa FITC-dextran, or adeno-associated virus (AAV) particles demonstrates that this bilayer model provides similar or greater barrier function than Transwell assays. Comparison of AAV serotypes indicated that AAV-PHP.eB can cross the human BBB in vitro, and at higher efficiency than AAV9. Additionally, RNAi knockdown and virus capsid protein binding assays show that AAV-PHP.eB delivery is facilitated by receptor protein lymphocyte antigen-6E (LY6E) in humans. This in vitro model system uses a miniaturized chip to enable high-throughput investigations of AAV crossing efficiency in the BBB, and provides strong initial evidence that human LY6E mediates AAV-PHP.eB crossing the BBB.

Genome-wide identification, evolutionary and expression analyses of LEA gene family in peanut (Arachis hypogaea L.).

BACKGROUND: Late embryogenesis abundant (LEA) proteins are a group of highly hydrophilic glycine-rich proteins, which accumulate in the late stage of seed maturation and are associated with many abiotic stresses. However, few peanut LEA genes had been reported, and the research on the number, location, structure, molecular phylogeny and expression of AhLEAs was very limited. RESULTS: In this study, 126 LEA genes were identified in the peanut genome through genome-wide analysis and were further divided into eight groups. Sequence analysis showed that most of the AhLEAs (85.7%) had no or only one intron. LEA genes were randomly distributed on 20 chromosomes. Compared with tandem duplication, segmental duplication played a more critical role in AhLEAs amplication, and 93 segmental duplication AhLEAs and 5 pairs of tandem duplication genes were identified. Synteny analysis showed that some AhLEAs genes come from a common ancestor, and genome rearrangement and translocation occurred among these genomes. Almost all promoters of LEAs contain ABRE, MYB recognition sites, MYC recognition sites, and ERE cis-acting elements, suggesting that the LEA genes were involved in stress response. Gene transcription analyses revealed that most of the LEAs were expressed in the late stages of peanut embryonic development. LEA3 (AH16G06810.1, AH06G03960.1), and Dehydrin (AH07G18700.1, AH17G19710.1) were highly expressed in roots, stems, leaves and flowers. Moreover, 100 AhLEAs were involved in response to drought, low-temperature, or Al stresses. Some LEAs that were regulated by different abiotic stresses were also regulated by hormones including ABA, brassinolide, ethylene and salicylic acid. Interestingly, AhLEAs that were up-regulated by ethylene and salicylic acid showed obvious subfamily preferences. Furthermore, three AhLEA genes, AhLEA1, AhLEA3-1, and AhLEA3-3, which were up-regulated by drought, low-temperature, or Al stresses was proved to enhance cold and Al tolerance in yeast, and AhLEA3-1 enhanced the drought tolerance in yeast. CONCLUSIONS: AhLEAs are involved in abiotic stress response, and segmental duplication plays an important role in the evolution and amplification of AhLEAs. The genome-wide identification, classification, evolutionary and transcription analyses of the AhLEA gene family provide a foundation for further exploring the LEA genes' function in response to abiotic stress in peanuts.

Recombinase-Aided Amplification Combined with Lateral Flow (LF-RAA) Assay for Rapid AAV Genome Detection.

Adeno-associated virus (AAV) is a versatile gene vector that is widely used in mammalian research. In basic studies and large-scale AAV production, genetic testing is ubiquitous and routine polymerase chain reaction (PCR)-based tests limit the efficiency due to the labor-intensive and time-consuming requirements of thermal cycling. This study introduces an assay based on recombinase-aided amplification combined with lateral flow (LF-RAA), which can quickly and accurately detect the AAV genome, thus improving the efficiency of AAV research and production. This application is the first use of an RAA approach to AAV genome detection. In this point-of-care testing (POCT) detection platform, the RAA reaction and LF readout are integrated into a user-friendly microfluidic chip that can be applied without advanced technical training. The LF-RAA chip provides high sensitivity, with a limit of detection of 10 copies/µL, and generates results quickly, and it only needs to be incubated for 10 min at a constant temperature, that is, 39 °C. Results are visualized on the LF Dipstick, and detection results are reliable, validated with 100% accuracy in 47 laboratory-produced recombination adeno-associated virus (rAAV) samples carrying target genes from several different viruses. The LF-RAA assay is applicable in AAV research and production processes requiring genome identification.

Traditional Chinese medicine in the treatment of nonalcoholic steatohepatitis.

Nonalcoholic steatohepatitis (NASH) is a common chronic liver disease in clinical practice. It has been considered that NASH is one of the main causes of chronic liver disease, cirrhosis and carcinoma. The mechanism of the NASH progression is complex, including lipid metabolism dysfunction, insulin resistance, oxidative stress, inflammation, apoptosis, fibrosis and gut microbiota dysbiosis. Except for lifestyle modification and bariatric surgery, there has been no pharmacological therapy that is being officially approved in NASH treatment. Traditional Chinese medicine (TCM), as a conventional and effective therapeutic strategy, has been proved to be beneficial in treating NASH in numbers of studies. In the light of this, TCM may provide a potential therapy for treating NASH. In this review, we summarized the associated mechanisms of action TCM treating NASH in preclinical studies and systematically analysis the effectiveness of TCM treating NASH in current clinical trials.

Genome-wide identification and evolutionary analysis of RLKs involved in the response to aluminium stress in peanut.

BACKGROUND: As an important cash crop, the yield of peanut is influenced by soil acidification and pathogen infection. Receptor-like protein kinases play important roles in plant growth, development and stress responses. However, little is known about the number, location, structure, molecular phylogeny, and expression of RLKs in peanut, and no comprehensive analysis of RLKs in the Al stress response in peanuts have been reported. RESULTS: A total of 1311 AhRLKs were identified from the peanut genome. The AhLRR-RLKs and AhLecRLKs were further divided into 24 and 35 subfamilies, respectively. The AhRLKs were randomly distributed across all 20 chromosomes in the peanut. Among these AhRLKs, 9.53% and 61.78% originated from tandem duplications and segmental duplications, respectively. The ka/ks ratios of 96.97% (96/99) of tandem duplication gene pairs and 98.78% (646/654) of segmental duplication gene pairs were less than 1. Among the tested tandem duplication clusters, there were 28 gene conversion events. Moreover, all total of 90 Al-responsive AhRLKs were identified by mining transcriptome data, and they were divided into 7 groups. Most of the Al-responsive AhRLKs that clustered together had similar motifs and evolutionarily conserved structures. The gene expression patterns of these genes in different tissues were further analysed, and tissue-specifically expressed genes, including 14 root-specific Al-responsive AhRLKs were found. In addition, all 90 Al-responsive AhRLKs which were distributed unevenly in the subfamilies of AhRLKs, showed different expression patterns between the two peanut varieties (Al-sensitive and Al-tolerant) under Al stress. CONCLUSIONS: In this study, we analysed the RLK gene family in the peanut genome. Segmental duplication events were the main driving force for AhRLK evolution, and most AhRLKs subject to purifying selection. A total of 90 genes were identified as Al-responsive AhRLKs, and the classification, conserved motifs, structures, tissue expression patterns and predicted functions of Al-responsive AhRLKs were further analysed and discussed, revealing their putative roles. This study provides a better understanding of the structures and functions of AhRLKs and Al-responsive AhRLKs.

Type 1 narcolepsy without cataplexy and treatment progress: a case report.

Narcolepsy is a sleep disorder with the main manifestations of excessive daytime sleepiness, cataplexy, sleep paralysis, sleep hallucinations, and nighttime sleep disturbance. It is still not fully recognized by clinicians, and many patients are often misdiagnosed with epilepsy, syncope, or mental disorders. In the present study, we report the first case of narcolepsy diagnosed at the district, with a complete medical history, objective examinations, and cerebrospinal fluid and hematological tests, but no cataplexy. Mutiple sleep latentcy test (MSLT) showed that the average sleep latency was 3.1 min, and abnormal REM sleep episodes were detected in 4 naps. The average REM latency was 1.3 min. We review the knowledge and researches on this disease in Mainland China in the past 10 years. Data from China in 2014 showed a significant increase in the incidence of narcolepsy in 2011 after the H1N1 epidemic in China in 2009. Despite the low incidence rate, diagnosis of narcolepsy is still confusing and needs clinicians' attention. Whether the incidence of narcolepsy may increase after covid19 remains to be observed. So far, there is no clear evidence to support immunotherapy. In conclusion, further studies are needed to verify more treatments and improve the patient's life quality.

Xiaoyaosan (Tiaogan-Liqi therapy) protects peritoneal macrophages from corticosterone-induced stress by regulating the interaction between glucocorticoid receptor and ABCA1.

BACKGROUND: Previous studies have reported that Xiaoyaosan (XYS), Tiaogan-Liqi therapy, has a protective function in depressive disorder, and can regulate body weight and corticosterone (CORT) level. However, little is known about the effect of XYS in treating atherosclerosis. This study aimed to explore the influence XYS on macrophage foam cell formation and related mechanism. METHODS: Rat peritoneal macrophages (PMs) were separated and stimulated with CORT and oxidized low density lipoprotein (ox-LDL). The serum was obtained from rats treated with different doses of XYS and was added into the medium for macrophages. Then, the cell activity and lipid content of PMs were measured through Cell Counting Kit-8 (CCK-8) assay and oil red staining, respectively. The expressions of glucocorticoid receptor (GR), ATP binding cassette subfamily A member 1 (ABCA1), and heat shock protein 90 (HSP90) were detected. In addition, overexpression of GR and ABCA1 was performed and the effect on XYS treatment was subsequently assessed. RESULTS: The CCK-8 assay showed the serum increased cell activity of CORT-induced stress PMs in a XYS dose-dependent manner. Oil red staining and enzyme-linked immunosorbent assay (ELISA) showed that the serum decreased lipids of PMs. In the XYS treated groups, HSP90 protein was decreased and protein levels of ABCA1 and GR were increased in cytoplasm, while GR protein in nucleus was decreased. Co-immunoprecipitation (Co-IP) assay indicated GR might interact with HSP90 and be involved with the function of XYS. Furthermore, overexpression of GR attenuated the protective function of XYS on CORT-induced stress in PMs, while overexpression of ABCA1 enhanced that. CONCLUSIONS: This study denoted that XYS could protect PMs from CORT-induced stress by regulating the interaction of GR and ABCA1, which might contribute to the treatment of atherosclerosis.

Copy Number Variation of the PIGY Gene in Sheep and Its Association Analysis with Growth Traits.

Copy number variation (CNV) is a type of genomic variation with an important effect on animal phenotype. We found that the PIGY gene contains a 3600 bp copy number variation (CNV) region located in chromosome 6 of sheep (Oar_v4.0 36,121,601-36,125,200 bp). This region overlaps with multiple quantitative trait loci related to phenotypes like muscle density and carcass weight. Therefore, in this study, the copy number variation of the PIGY gene was screened in three Chinese sheep breeds, namely, Chaka sheep (CKS, May of 2018, Wulan County, Qinghai Province, China), Hu sheep (HS, May of 2015, Mengjin County, Henan Province, China), and small-tailed Han sheep (STHS, May of 2016, Yongjing, Gansu Province, China). Association analyses were performed on the presence of CNV and sheep body size traits. We used real-time quantitative PCR (qPCR) to detect the CNV for association analysis. According to the results, the loss-type CNV was more common than other types in the three breeds (global average: loss = 61.5%, normal = 17.5%, and gain = 21.0%). The association analysis also showed significant effects of the PIGY gene CNV on body weight, chest circumference, and circumference of the cannon bone of sheep. Sheep with gain-type CNV had better growth traits than those with other types. The results indicate a clear relationship between the PIGY gene CNV and growth traits of sheep, suggesting the use of CNV as a new molecular breeding marker.

Xiaoyaosan prevents atherosclerotic vulnerable plaque formation through heat shock protein/glucocorticoid receptor axis-mediated mechanism.

Atherosclerosis is a metabolic and chronic inflammatory disease caused by deposition of lipoproteins in arteries. However, the diagnostic drug and the mechanism for this vascular disease are less studied. In the present study, atherosclerosis model was developed using apolipoprotein E-deficient mice that was treated with long-term high-fat food and chronic stresses. Xiaoyaosan (XYS) and glucocorticoid receptor (GR) antagonist RU 38486 were orally administrated to the mice. The change of aortic root vessels was observed by histological analysis. The results indicate that high-fat food coupled with chronic stress induced atherosclerosis in mice model, with plaque formation in the entire aortas foam cells and macrophage infiltration in aortic tissues. However, XYS granules inhibited the development of atherosclerotic lesion, with down-regulation of GC, TC, TG, HDL-C, ox-LDL, LDL-C, IFN-γ, IL-6, IL-1ß, and TNF-α measured by ELISA method; XYS inhibited the expressions of GR, CD36, HSP27/60/90, and induced ABCA1 in atherosclerotic mice, which was measured by qPCR and Western blot, which showed similar effect as positive control RU 38486 did. The interaction between HSP90-GR complexes and CD36 was validated in atherosclerotic mice. Our results inferred that the HSP/GR complex-mediated CD36 axis was involved in the regulation of atherosclerosis development in mice verified by Co-IP assay, EMSA, and Chip-PCR. These findings not only provide the potential therapeutic value of Xiaoyaosan for atherosclerotic vulnerable plaque but also brought forth a novel strategy for preventing the formation and treatment of atherosclerotic vulnerable plaques through the elucidated mechanism of XYS on vulnerable plaque.

Increased Spot Urine Albumin-to-Creatinine Ratio and Stroke Incidence: A Systematic Review and Meta-Analysis.

OBJECTIVE: This study aimed to clarify the association between an increased spot urine albumin-to-creatinine ratio (UACR) and the risk of stroke. METHODS: We performed a systematic review and meta-analysis of cohort studies, case-control studies, and ancillary data randomized controlled trials (RCTs), which were treated as cohorts in this study, and estimated the association between albuminuria, as measured with the UACR, and the risk of stroke. We performed a comprehensive search of PubMed, Embase, and the Cochrane Library and conducted a systematic review and cumulative meta-analysis of cohort studies with a cross-sectional with prospective design in which stroke incidence was reported and the baseline UACR was measured. Ancillary data from RCTs were also included as part of the cohort study. We studied the characteristics of the participants, quality scores and risk ratios (RR, with confidence intervals, CI) of stroke associated with normal and high UACRs, and we synthesized the data via a meta-analysis. RESULTS: Twelve eligible studies including a total of 32,888 participants and 3,944 cases of stroke were identified. A high UACR (>30 mg/mmol) increased the risk of stroke by 1.67 times (RR: 1.67, 95% CI: 1.49-1.86, P<0.001 I2 = 26%). The results were not different between Asian and non-Asian patients (RR: 1.64, 95% CI: 1.41-1.91, P<0.001, I2 = 23% compared with RR: 1.67, 95% CI: 1.50-1.85, P<0. 00, I2 = 39%) or between subgroups classified by old age (RR: 1.61, 95% CI: 1.39-1.88, P<0.001, I2 = 34% compared with RR: 1.68, 95% CI: 1.52-1.87, P<0.001, I2 = 13%). A sensitivity analysis did not significantly change the results. CONCLUSION: The incidence of stroke increased significantly in the high UACR group compared with the normal UACR group. The UACR could be a clinical addition for the early indication of high-risk stroke patients.

Pollen Typhae Total Flavone Inhibits Endoplasmic Reticulum Stress-Induced Apoptosis in Human Aortic-Vascular Smooth Muscle Cells through Down-Regulating PERK-eIF2α-ATF4-CHOP Pathway.

OBJECTIVE: To test the hypothesis that the inhibition of endoplasmic reticulum (ER) stress-induced apoptosis in oxidized low-density lipoproteins (ox-LDL)-induced human aortic-vascular smooth muscle cells (HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase (PERK)-eukaryotic translation initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4)-CCAAT/enhancer binding protein homologous protein (CHOP) signaling pathway by Pollen Typhae total flavone (PTF). METHODS: Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group (70 µg/mL high ox-LDL), an HPTF group (70 µg/mL high ox-LDL+500 µg/mL PTF), an MPTF group (70 µg/mL high ox-LDL+250 µg/mL PTF), and a LPTF group (70 µg/mL high ox-LDL+100 µg/mL PTF) in the first part; and a normal control group, an ox-LDL group (70 µg/mL high ox-LDL), an MPTF group (70 µg/mL high ox-LDL+250 µg/mL PTF), a shRNA group (transducted with PERK shRNA lentiviral particles), a scramble shRNA group (transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group (250 µg/mL PTF+70 µg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group (70 µg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their mRNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was applied to test cell viability, and the level of apoptosis was monitored by flow cytometry. RESULTS: The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and shRNA groups. Moreover, the ox-LDL group had increased protein and mRNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK, eIF2α, ATF4 and CHOP, which were attenuated in PTF treatment groups and shRNA groups. CONCLUSIONS: The apoptosis induced by ox-LDL had a strong relation to ER stress. The protective effect of PTF on ER stressinduced apoptosis was associated with inhibition of the PERK-eIF2α-ATF4-CHOP pathway, which might be a potential therapeutic strategy for enhancing the stability of atherosclerotic plaques.

A Dual-Label Time-Resolved Fluorescence Immunoassay for the Simultaneous Determination of Cardiac Troponin T and Myoglobin.

The aim of this study was to establish a dual-label time-resolved fluorescence immunoassay (TRFIA) for the simultaneous determination of cardiac troponin T (cTnT) and myoglobin (MYO) for the early diagnosis of acute myocardial infarction. The sandwich immunoassay was used to detect the concentration of cTnT and MYO in serum. cTnT and MYO in serum were captured by anti-cTnT and anti-MYO antibodies immobilized on microtiter wells and then banded together with another anti-cTnT and anti-MYO labeled with europium(III) Sm3+ and samarium(III) Eu3+ chelates, followed by fluorescence measurement using time-resolved fluorometry. The performance of this TRFIA was evaluated using the clinical serum and compared with the commercial assays. The linear correlation coefficients ( R2) of the cTnT and MYO standard curves were 0.9993 and 0.9995, respectively. The sensitivity for cTnT detection was 2.21 pg/mL (linear dynamic range was 3.24-963.71 pg/mL), and the average recovery was 100.57%. The sensitivity for MYO detection was 3.24 ng/mL (linear dynamic range was 4.95-976.85 ng/mL), and the average recovery was 99.79%. High correlation coefficients ( R2) were obtained between the commercial assays and dual-label TRFIA ( R2 = 0.999). The present dual-label TRFIA has high sensitivity, specificity, and accuracy in clinical sample analysis. It is a good alternative to the single-label diagnostic methods.

A dual-label time-resolved fluorescence immunoassay (TRFIA) for screening of Coronary atherosclerosis based on simultaneous detection of Lp-PLA2 and HsCRP.

Atherosclerosis is the underlying cause of most coronary events. The conventional method for coronary atherosclerosis detection is morphological examination or coronary arterionyraphy. These methods are complex and time-consuming. In this study a two-step dual-label TRFIA was developed for the simultaneous detection of Lp-PLA2 and hsCRP in a single run. The performance of this assay was first evaluated using clinical serum samples, and then compared with commercialized kits. The sensitivity of this assay for Lp-PLA2 detection was 1ng/mL (dynamic range, 0-1000U/L), and the sensitivity for hsCRP detection was 1mg/L (dynamic range, 1-1000mg/L). High correlation coefficients (R) were obtained between the present dual-label TRFIA and commercially available kits(R=0.99 for LP-PLA2 and hsCRP). The present dual-label TRFIA has high sensitivity, specificity, and accuracy in clinical sample analysis. It is a good alternative to the single-label diagnostic methods.

[Effect of Sini decoction on function of hypothalamic-pituitary-adrenal axis in patients with sepsis].

OBJECTIVE: To investigate the effects of Sini decoction on function of hypothalamic-pituitary-adrenal axis in patients with sepsis. METHODS: A prospective single-blind randomized controlled trial was conducted. 60 septic patients were divided into three groups with the method of random number table, 20 cases in the control group, 20 in the Chinese herb group, and 20 in corticoid group. All of them received routine treatment. Patients in Chinese herb group were given Sini decoction in addition (decoction of monkshood 15 g, dried ginger 15 g, honey-fried licorice 10 g) 100 mL/d orally or by nasal feeding, while patients in corticoid group were given hydrocortisone 200 mg/d intravenously instead, both for 7 days. Before the treatment, 3 days and 14 days after treatment, blood was collected to determine the levels of adrenocorticotropic hormone (ACTH) and cortisol, and the result of ACTH stimulating test was observed. At the same time, acute physiology and chronic health evaluation II (APACHEII) score was recorded, and 3-day shock recovery rate and 28-day death rate were also compared among these groups. RESULTS: None of the three groups showed different result in ACTH stimulating test (χ(2)=1.101, P=0.605). ACTH in three groups was gradually decreased. Compared with that before treatment, ACTH in Chinese herb group and corticoid groups began to decrease obviously on day 3 (29.90±3.31 ng/L vs. 33.10±3.31 ng/L, 28.20±2.45 ng/L vs. 33.30±3.84 ng/L, both P<0.01), while in control group declined ACTH appeared later (on day 14) compared with before treatment (29.40±5.63 ng/L vs. 33.50±4.89 ng/L, P<0.05). No obvious difference in ACTH level was showed between the Chinese herb group and the cortical group (both P>0.05). Cortisol level in both Chinese herb and cortical groups showed a raise-fall biphase trend while there was no change in the control. The cortical levels on day 3 in Chinese herb and cortical groups were much higher than that before treatment (343.04±31.20 µg/L vs. 294.70±42.10 µg/L, 331.25±42.80 µg/L vs. 280.36±38.10 µg/L, both P<0.01) and that of control group (291.61±41.50 µg/L, both P<0.01), though no significant statistical difference was observed between two groups (both P>0.05). APACHEII score on day 14 in control, Chinese herb and cortical groups was significantly lower than that before treatment (16.8±5.1 vs. 20.1±4.3, 13.4±3.2 vs. 18.3±3.8,15.1±2.5 vs. 19.5±4.0, all P<0.01), and the score was much lower in Chinese herb group comparing with that of control group (P<0.05). No statistical difference was observed among control, Chinese herb and cortical groups in lowering 28-day death rate [35.0% (7/20), 25.0% (5/20), 20.0% (4/20)] and improving 3-day shock recovery rate [40.0% (8/20), 70.0% (14/20), 60.0% (12/20), all P>0.05]. CONCLUSIONS: Sini decoction could elevate cortisol while lower ACTH at the early stage of sepsis. Sini decoction could also effectively improve symptoms and hypothalamic-pituitary-adrenal axis function in septic patients without affecting death rate.