Effect of reactor operation modes on mitigating antibiotic resistance genes (ARGs) and methane production from hydrothermally-pretreated pig manure.

Numerous efforts have been made to enhance the performance of anaerobic digestion (AD) for accelerating renewable energy generation, however, it remains unclear whether the intensified measures could enhance the proliferation and transmissions of antibiotic resistance genes (ARGs) in the system. This study assessed the impact of an innovative pig manure AD process, which includes hydrothermal pretreatment (HTP) and a two-stage configuration with separated acidogenic and methanogenic phases, on biomethane (CH4) production and ARGs dynamics. Results showed that HTP significantly increase CH4 production from 0.65 to 0.75 L/L/d in conventional single-stage AD to 0.82 and 0.91 L/L/d in two-stage AD. This improvement correlated with a rise in the relative abundance of Methanosarcina, a key methanogenesis microorganism. In the two-stage AD, the methanogenic stage offered an ideal environment for methanogens growth, resulting in substantially faster and higher CH4 production by about 10% compared to single-stage AD. Overall, the combined use of HTP and the two-stage AD configuration enhanced CH4 production by 40% compared to traditional single-stage AD. The abundance and diversity of ARGs were significantly reduced in the acidogenic reactors after HTP. However, the ARGs levels increased by about two times in the following methanogenesis stage and reached similar or higher levels than in single stage AD. The erm(F), erm(G), ant(6)-Ia, tet(W), mef(A) and erm(B) were the six main ARGs with significant differences in relative abundances in various treatments. The two-stage AD mode could better remove sul2, but it also had a rebound which elevated the risk of ARGs to the environment and human health. Network analysis identified pH and TVFAs as critical factors driving microbial communities and ARG proliferation in the new AD process. With the results, this study offers valuable insights into the trade-offs between AD performance enhancement and ARG-related risks, pinpointing essential areas for future research and practical improvements.

Micropollutant rejection by nanofiltration membranes: A mini review dedicated to the critical factors and modelling prediction.

Nanofiltration (NF) membranes, extensively used in advanced wastewater treatment, have broad application prospects for the removal of emerging trace organic micropollutants (MPs). The treatment performance is affected by several factors, such as the properties of NF membranes, characteristics of target MPs, and operating conditions of the NF system concerning MP rejection. However, quantitative studies on different contributors in this context are limited. To fill the knowledge gap, this study aims to assess critical impact factors controlling MP rejection and develop a feasible model for MP removal prediction. The mini-review firstly summarized membrane pore size, membrane zeta potential, and the normalized molecular size (λ = rs/rp), showeing better individual relationships with MP rejection by NF membranes. The Lindeman-Merenda-Gold model was used to quantitatively assess the relative importance of all summarized impact factors. The results showed that membrane pore size and operating pressure were the high impact factors with the highest relative contribution rates to MP rejection of 32.11% and 25.57%, respectively. Moderate impact factors included membrane zeta potential, solution pH, and molecular radius with relative contribution rates of 10.15%, 8.17%, and 7.83%, respectively. The remaining low impact factors, including MP charge, molecular weight, logKow, pKa and crossflow rate, comprised all the remaining contribution rates of 16.19% through the model calculation. Furthermore, based on the results and data availabilities from references, the machine learning-based random forest regression model was trained with a relatively low root mean squared error and mean absolute error of 12.22% and 6.92%, respectively. The developed model was then successfully applied to predict MPs' rejections by NF membranes. These findings provide valuable insights that can be applied in the future to optimize NF membrane designs, operation, and prediction in terms of removing micropollutants.

Whole genome sequencing reveals novel genetic mutations of Helicobacter pylori associating with resistance to clarithromycin and levofloxacin.

BACKGROUND: Detection of mutations in one or a couple of genes may not provide enough data or cover all the genomic DNA variance related to antibiotic resistance of Helicobacter pylori to clarithromycin (CLA) and levofloxacin (LVX). We aimed to perform whole genome sequencing to explore novel antibiotic resistance-related genes to increase predictive accuracy for future targeted sequencing tests. METHODS: Gastric mucosal biopsies were taken during upper endoscopy in 27 H. pylori-infected patients. According to culture-based antibacterial susceptibility test, H. pylori strains were divided into three groups, with nine strains in each group: CLA single-drug resistance (group C), LVX single-drug resistance (group L), and strains sensitive to all antibacterial drugs (group S). Based on whole genome sequencing with group S being the control, group C and group L group-specific single nucleotide variants and amino acid mutations were screened, and potential candidate genes related to CLA and LVX resistance were identified. RESULTS: The median age of study subjects was 35 years (IQR: 31-40), and 17 (63.0%) were male. All nine CLA-resistant strains had A2143G mutations in 23S rRNA, while none of nine sensitive strains had the mutation. Six of nine strains in group L and six of nine strains in group S had 87th or 91st mutation in gyrA. After comparing sequencing data of strains among the three groups, we identified five mutated positions belonging to four genes related to CLA resistance, and 31 mutated positions belonging to 20 genes related to LVX resistance. Novel genetic mutations were detected for CLA resistance (including fliJ and clpX) and LVX resistance (including fliJ, cheA, hemE, Val360Ile, and HP0568). Missense mutations in fliJ and cheA gene were mainly involved in chemotaxis and flagellar motility to facilitate bacterial escape of antibiotics, while the functions of other novel gene mutations underpinning antibiotic resistance remain to be investigated. CONCLUSION: Whole genome sequencing detected potential novel genetic mutations conferring resistance of H. pylori to CLA and LVX including fliJ and cheA. Further studies to correlate these findings with treatment outcome should be performed.

Vonoprazan-based triple and dual therapy versus bismuth-based quadruple therapy for Helicobacter pylori infection in China: a three-arm, randomised clinical trial protocol.

BACKGROUND: Helicobacter pylori infection and associated diseases are a growing global public health issue. H. pylori infection is the major cause of gastric cancer, over 90% of duodenal ulcers, and over 70% of gastric ulcers. The infection rate of H. pylori is approximately 50%, and approximately 50% of new cases of gastric cancer worldwide occur in China. Bismuth (BI)-based quadruple therapy is recommended as the first-line treatment for H. pylori in China. Vonoprazan (VPZ), a new potassium-competitive acid blocker that can inhibit gastric acid secretion more effectively than proton pump inhibitors (PPIs), has been combined with antibiotics to effectively eradicate H. pylori. In this study, we compared the efficacy and safety of two VPZ-based therapies with that of BI-based therapy for H. pylori treatment. METHODS: A three-armed randomised controlled trial (RCT) is being conducted in Shenzhen, with 327 participants recruited from the Gastroenterology Clinic of the University of Hong Kong-Shenzhen Hospital. Patients were diagnosed with H. pylori infection based on a positive 13C-urea breath test (UBT). Patients are kept naïve to their treatment and are randomly assigned in a 1:1:1 ratio to either VPZ-based triple, VPZ-based dual, or BI-based quadruple therapy for 14 days. All groups are subjected to follow-up evaluations of safety, adverse drug reactions, and clinical variables in the first, second, and fourth weeks after treatment. Successful eradication is confirmed by a negative 13C-UBT six weeks after treatment. If initial treatment fails, (1) those patients are turned to another regimen, or (2) a drug resistance test is conducted, after which an individualised treatment regimen shall be prescribed according to antimicrobial susceptibility testing. The resulting data will be evaluated using intention-treat and a per-protocol analysis. DISCUSSION: This study is the a RCT aims to evaluate the efficacy and safety of 14-day VPZ-based triple and dual therapies in comparison with BI-based quadruple therapy. The outcomes of this study may allow treatment recommendations and update drug instructions in China. TRIAL REGISTRATION: Chinese Clinical Trial Registry (No. ChiCTR2200056375). Registered on February 4, 2022, https://www.chictr.org.cn/showproj.aspx?proj=141314.

Assessing the efficacy and mechanisms of glycol-contaminated water treatment through floating treatment wetlands.

The growing concerns surrounding water pollution and the degradation of ecosystems worldwide have led to an increased use of nature-based solutions (NbSs). This study assessed the feasibility of using floating treatment wetlands (FTWs) as an NbS to treat propylene glycol-contaminated water and quantitatively investigated different removal pathways. With an environmentally relevant concentration of propylene glycol (1,250 mg/L), FTWs containing Acorus calamus and mixed species demonstrated the highest average glycol mass removal efficacy (99%), followed by Carex acutiformis (98%), Juncus effusus (93%), and the control group without plants (10%) after 1 week. Additional mesocosm-scale experiments with varying FTW configurations, including surface coverage to reduce evaporation and photodegradation processes, and the addition of antibiotics to inhibit microbial activity, were conducted to quantify glycol removal pathways. Mass balance analysis results revealed that microbial biodegradation (33.3-39.7%) and plant uptake (37.9-45.2%) were the primary pathways for glycol removal. Only 15.5-19.5% of the glycol removal via evaporation and photodegradation was accounted in this study, which may be attributed to the mesocosm experimental setup (static water and no wind). Aligned with the broader discussion regarding biodiversity improvements and carbon storage capacity, this study demonstrated that FTWs are an environmentally friendly and effective NbS for addressing glycol-contaminated water.

Revealing the link between evolution of electron transfer capacity of humic acid and key enzyme activities during anaerobic digestion.

Humic acid (HA) is an important active compound formed during anaerobic digestion process, with a complex structure and dynamic electron transfer capacity (ETC). However, the mechanisms by which these macromolecular organic compounds dynamically interact with the microbial anaerobic digestion process at different operating temperatures are still unclear. In this study, the link between the evolution of the ETC of HAs and the microbial community under mesophilic and thermophilic conditions was investigated. The results showed an increasing trend in the ETC of HAs in both mesophilic (671-1479 µmol gHA-1) and thermophilic (774-1506 µmol gHA-1) anaerobic digestion (AD) until day 25. The ETC was positively correlated with the bacterial community of hydrolytic and acidogenic phases, but negatively correlated with the archaeal community of the methanogenic phase. Furthermore, the relationship between ETC and key enzyme activity was explored using a co-occurrence network analysis. HAs revealed a high potential to promote key enzyme activities during hydrolysis (amylase and protease) and acidification (acetate kinase, butyrate kinase, and phosphotransacetylase) while inhibiting the key enzyme activity in the methanogenic phase during the anaerobic digestion process. Moreover, HAs formed under thermophilic conditions had a greater influence on key enzyme activities than those formed under mesophilic conditions. This study advances our understanding of the mechanisms underlying the influence of HAs on anaerobic digestion performance.

Alternative Polyadenylation in response to temperature stress contributes to gene regulation in Populus trichocarpa.

BACKGROUND: Genome-wide change of polyadenylation (polyA) sites (also known as alternative polyadenylation, APA) is emerging as an important strategy of gene regulation in response to stress in plants. But little is known in woody perennials that are persistently dealing with multiple abiotic stresses. RESULTS: Here, we performed a genome-wide profiling of polyadenylation sites under heat and cold treatments in Populus trichocarpa. Through a comprehensive analysis of polyA tail sequences, we identified 25,919 polyA-site clusters (PACs), and revealed 3429 and 3139 genes shifted polyA sites under heat and cold stresses respectively. We found that a small proportion of genes possessed APA that affected the open reading frames; and some shifts were commonly identified. Functional analysis of genes displaying shifted polyA tails suggested that pathways related to RNA metabolism were linked to regulate the APA events under both heat and cold stresses. Interestingly, we found that the heat stress induced a significantly more antisense PACs comparing to cold and control conditions. Furthermore, we showed that a unique cis-element (AAAAAA) was predominately enriched downstream of PACs in P. trichocarpa genes; and this sequence signal was only absent in shifted PACs under the heat condition, indicating a distinct APA mechanism responsive to heat tolerance. CONCLUSIONS: This work provides a comprehensive picture of global polyadenylation patterns in response to temperatures stresses in trees. We show that the frequent change of polyA tail is a potential mechanism of gene regulation responsive to stress, which are associated with distinctive sequence signatures.

Development and validation of a 14-gene signature for prognosis prediction in hepatocellular carcinoma.

Worldwide, hepatocellular carcinoma (HCC) remains a crucial medical problem. Precise and concise prognostic models are urgently needed because of the intricate gene variations among liver cancer cells. We conducted this study to identify a prognostic gene signature with biological significance. We applied two algorithms to generate differentially expressed genes (DEGs) between HCC and normal specimens in The Cancer Genome Atlas cohort (training set included) and performed enrichment analyses to expound on their biological significance. A protein-protein interactions network was established based on the STRING online tool. We then used Cytoscape to screen hub genes in crucial modules. A multigene signature was constructed by Cox regression analysis of hub genes to stratify the prognoses of HCC patients in the training set. The prognostic value of the multigene signature was externally validated in two other sets from Gene Expression Omnibus (GSE14520 and GSE76427), and its role in recurrence prediction was also investigated. A total of 2000 DEGs were obtained, including 1542 upregulated genes and 458 downregulated genes. Subsequently, we constructed a 14-gene signature on the basis of 56 hub genes, which was a good predictor of overall survival. The prognostic signature could be replicated in GSE14520 and GSE76427. Moreover, the 14-gene signature could be applied for recurrence prediction in the training set and GSE14520. In summary, the 14-gene signature extracted from hub genes was involved in some of the HCC-related signalling pathways; it not only served as a predictive signature for HCC outcome but could also be used to predict HCC recurrence.

[Portable Pulse Detection System Based on IoT].

Aiming at the current situation of high cost, huge volume, complex operation and difficulty in real application of pulse analyzer, this study designs and implements a portable pulse detection system based on IoT. The design utilizes Raspberry Pi 3B+, STM32 series MCU and cloud server to collect, store, display and recognize pulse signals at CUN, GUAN and CHI. The system is small in size and low in cost, which can be connected with cloud server through network to make full use of resources. The experimental results show that the recognition accuracy of the main feature points of the pulse signal by the portable pulse analyzer is higher than 97%, which has a broad prospect of development and application.

High prevalence and risk factors of multiple antibiotic resistance in patients who fail first-line Helicobacter pylori therapy in southern China: a municipality-wide, multicentre, prospective cohort study.

BACKGROUND: We aimed to study the prevalence of secondary antibiotic resistance of Helicobacter pylori in southern China and its risk factors, particularly geographical and socio-economic factors. METHODS: This was a municipality-wide, multicentre, prospective cohort study involving five major hospitals. Patients aged ≥18 years who failed first-line bismuth-based quadruple anti-H. pylori therapy between September 2016 and February 2018 were recruited. Participants underwent upper gastrointestinal endoscopy with biopsy from the antrum and body for H. pylori culture and antimicrobial susceptibility testing for six antibiotics (clarithromycin, levofloxacin, metronidazole, amoxicillin, tetracycline and furazolidone). Patients with failure of H. pylori culture were excluded. Participants completed a questionnaire profiling 22 potential risk factors of H. pylori infection and antibiotic resistance, including medical, social, household and birthplace factors. RESULTS: A total of 1113 patients failed first-line therapy, with successful H. pylori culture in 791 (71.1%) [male = 433 (54.7%); median age = 43 years]. Secondary resistance rates of dual antibiotics (clarithromycin + metronidazole and levofloxacin + metronidazole) and triple antibiotics (clarithromycin + levofloxacin + metronidazole) were 34.0%, 38.7% and 17.8%, respectively. Risk factors for clarithromycin + metronidazole resistance were history of ≥2 courses of H. pylori therapies [adjusted OR (aOR) = 1.71; 95% CI = 1.17-2.54], ≥3 household members (aOR = 2.00; 95% CI = 1.07-3.90) and family history of gastric cancer (aOR = 1.85; 95% CI = 1.18-2.85). Risk factors for levofloxacin + metronidazole resistance were age ≥40 years (aOR = 1.94; 95% CI = 1.37-2.75), lower gross domestic product per capita (aOR = 0.29; 95% CI = 0.10-0.80) and higher number of doctors/10 000 population (aOR = 1.59; 95% CI = 1.07-2.39). A higher human development index was of borderline significance (aOR = 2.79; 95% CI = 0.97-8.70). CONCLUSIONS: The rates of secondary resistance of H. pylori to multiple antibiotics were high in southern China. Certain population-level risk factors were associated with levofloxacin + metronidazole resistance.

Identification of alternatively spliced gene isoforms and novel noncoding RNAs by single-molecule long-read sequencing in Camellia.

Direct single-molecule sequencing of full-length transcripts allows efficient identification of gene isoforms, which is apt to alternative splicing (AS), polyadenylation, and long non-coding RNA analyses. However, the identification of gene isoforms and long non-coding RNAs with novel regulatory functions remains challenging, especially for species without a reference genome. Here, we present a comprehensive analysis of a combined long-read and short-read transcriptome sequencing in Camellia japonica. Through a novel bioinformatic pipeline of reverse-tracing the split-sites, we have uncovered 257,692 AS sites from 61,838 transcripts; and 13,068 AS isoforms have been validated by aligning the short reads. We have identified the tissue-specific AS isoforms along with 6,373 AS events that were found in all tissues. Furthermore, we have analysed the polyadenylation (polyA) patterns of transcripts, and found that the preference for polyA signals was different between the AS and non-AS transcripts. Moreover, we have predicted the phased small interfering RNA (phasiRNA) loci through integrative analyses of transcriptome and small RNA sequencing. We have shown that a newly evolved phasiRNA locus from lipoxygenases generated 12 consecutive 21 bp secondary RNAs, which were responsive to cold and heat stress in Camellia. Our studies of the isoform transcriptome provide insights into gene splicing and functions that may facilitate the mechanistic understanding of plants.

Sustainable Chromium (VI) Removal from Contaminated Groundwater Using Nano-Magnetite-Modified Biochar via Rapid Microwave Synthesis.

This study developed a nano-magnetite-modified biochar material (m-biochar) using a simple and rapid in situ synthesis method via microwave treatment, and systematically investigated the removal capability and mechanism of chromium (VI) by this m-biochar from contaminated groundwater. The m-biochar was fabricated from reed residues and magnetically modified by nano-Fe3O4. The results from scanning electron microscopy (SEM) and X-ray diffraction (XRD) characterisations confirmed the successful doping of nano-Fe3O4 on the biochar with an improved porous structure. The synthesised m-biochar exhibited significantly higher maximum adsorption capacity of 9.92 mg/g compared with that (8.03 mg/g) of the pristine biochar. The adsorption kinetics followed the pseudo-second-order model and the intraparticle diffusion model, which indicated that the overall adsorption rate of Cr(VI) was governed by the processes of chemical adsorption, liquid film diffusion and intramolecular diffusion. The increasing of the pH from 3 to 11 significantly affected the Cr(VI) adsorption, where the capabilities decreased from 9.92 mg/g to 0.435 mg/g and 8.03 mg/g to 0.095 mg/g for the m-biochar and pristine biochar, respectively. Moreover, the adsorption mechanisms of Cr(VI) by m-biochar were evaluated and confirmed to include the pathways of electrostatic adsorption, reduction and complexation. This study highlighted an effective synthesis method to prepare a superior Cr(VI) adsorbent, which could contribute to the effective remediation of heavy metal contaminations in the groundwater.

CjPLE, a PLENA-like gene, is a potential regulator of fruit development via activating the FRUITFUL homolog in Camellia.

Fruit patterning involves the cooperation of multiple processes, including metabolic change, cell differentiation, and cell expansion. The FRUITFUL (FUL) and SHATTERPROOF1/2 (SHPs) MADS-box genes are master regulators directing fruit patterning in several eudicots. However, the regulatory mechanisms of the FUL-SHP network in different fruit types remain unclear. Here, we characterized the functions of an ortholog (CjPLE) of SHPs from Camellia japonica. We showed that CjPLE was predominantly expressed in stamen and carpel tissues during the early stage of floral development and that transcripts were abundant in the pericarp tissues during fruit development. The ectopic expression of CjPLE in Arabidopsis caused enhanced development of the carpels, whereas no defects in floral identity were observed. To investigate the downstream targets of CjPLE, overexpression transformants were analysed through a callus transformation system in Camellia azalea. We examined the expression levels of potential downstream target genes and found that two previously identified APETALA1-like genes (CjAPL1/2) were significantly up-regulated. We showed that CjPLE directly bound to the CArG motifs in the promoter region of CjAPL1 (the FUL ortholog). Taken together, our results reveal a possible positive regulation of FUL by SHP in the control of fruit development in Camellia.

[C_(19)-diterpenoid alkaloids from Aconitum austroyunnanense].

Eight C_(19)-diterpenoid alkaloids( 1-8) were isolated from the ethyl acetate soluble fraction of 95% ethanol extract of the ground roots of Aconitum austroyunnanense through various column chromatographies on silica gel,ODS,Sephadex LH-20 and MCI gel.Their structures were elucidated as 14α-benzoyloxy-13ß,15α-dihydroxy-1α,6α,8ß,16ß,18-pentamethoxy-19-oxoaconitan( 1),N-deethylaconitine( 2),spicatine B( 3),leucanthumsine A( 4),acofamine B( 5),macrorhynine B( 6),aconitilearine( 7),and ambiguine( 8) based on their chemical and physicochemical properties and spectroscopic data. Compound 1 was a new compound and alkaloids 2-8 were isolated from this plant for the first time. Some isolated alkaloids were tested in vitro for cytotoxic potential by employing the MTT method. As a result,alkaloid 1 exhibited weak cytotoxic activity against three tested tumor cell lines( A-549,He La,and Hep G2) with IC_(50) values less than 20 µmol·L~(-1).

Comparative genomics analysis reveals gene family expansion and changes of expression patterns associated with natural adaptations of flowering time and secondary metabolism in yellow Camellia.

Yellow-flowering species are unique in the genus Camellia not only for their bright yellow pigments but also the health-improving substances in petals. However, little is known regarding the biosynthesis pathways of pigments and secondary metabolites. Here, we performed comparative genomics studies in two yellow-flowered species of the genus Camellia with distinctive flowering periods. We obtained 112,190 and 89,609 unigenes from Camellia nitidissima and Camellia chuongtsoensis, respectively, and identified 9547 gene family clusters shared with various plant species and 3414 single-copy gene families. Global gene expression analysis revealed six comparisons of differentially expressed gene sets in different developmental stages of floral bud. Through the identification of orthologous pairs, conserved and specific differentially expressed genes (DEGs) between species were compared. Functional enrichment analysis suggested that the gibberellin (GA) biosynthesis pathway might be related to the alteration of flowering responses. Furthermore, the expression patterns of secondary metabolism pathway genes were analyzed between yellow- and red-flowered Camellias. We showed that the key enzymes involved in glycosylation of flavonoids displayed differential expression patterns, indicating that the direct glycosylation of flavonols rather than anthocyanins was pivotal to coloration and health-improving metabolites in the yellow Camellia petals. Finally, the gene family analysis of UDP-glycosyltransferases revealed an expansion of group C members in C. nitidissima. Through comparative genomics analysis, we demonstrate that changes of gene expression and gene family members are critical to the variation of natural traits. This work provides valuable insights into the molecular regulation of trait adaptations of floral pigmentation and flowering timing.

Switching Harmful Algal Blooms to Submerged Macrophytes in Shallow Waters Using Geo-engineering Methods: Evidence from a 15N Tracing Study.

Switching the dominance from algae to macrophytes is crucial for lake management of human-induced eutrophication. Nutrients from algal sources can be utilized in the process of transition from algal blooms to macrophytes, thereby mitigating eutrophication. However, this process rarely occurs in algal bloom dominated waters. Here, we examined the hypothesis that the transition of algal blooms to macrophytes and the transfer of nutrients from algae at different temperatures (8 and 25 °C) can be facilitated by using a geo-engineering method. The results showed that the combination of flocculation and capping treatment could not only remove Microcystis aeruginosa blooms from eutrophic waters but also facilitate algal decomposition and incorporation into a submerged macrophyte ( Potamogeton crispus) biomass. The flocculation-capping treatment could trigger algal cell lysis. As compared with the control groups, the photosynthesis and respiration rate of algae were inhibited and chlorophyll-a (Chl- a) concentrations were significantly reduced in the flocculation-capping treatment groups. The 15N tracing study revealed that 3.3% and 34.8% of algae-derived nitrogen could be assimilated by Potamogeton crispus at 8 and 25 °C, respectively. The study demonstrated that the flocculation-capping method can facilitate the switchover from algae- to the macrophyte-dominated state, which is crucial for restoring the aquatic ecosystem.

Estimation of the incidence of severe fever with thrombocytopenia syndrome in high endemic areas in China: an inpatient-based retrospective study.

BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is a severe viral disease caused by SFTSV. It is important to estimate the rate of missed SFTS diagnosis and to further understand the actual incidence in high endemic areas in China. METHODS: This study was conducted in two high SFTS endemic provinces in 2015. Patients hospitalized in 2014 or within 1 year before investigation were selected after considering their clinical manifestations, specifically, fever, platelet, and white blood cell. During retrospective investigation, sera were collected to detect SFTSV antibodies to assess SFTSV infection. To further understand SFTSV infection, acute phase sera were detected; SFTSV infection rate among a healthy population was also investigated to determine the basic infection level. RESULTS: In total, 246 hospitalized cases were included, including 83 cases (33.7%) with fever, thrombocytopenia and leukopenia, 38 cases (15.4%) with fever and thrombocytopenia but without leukopenia, and 125 cases (50.8%) without fever but with thrombocytopenia and leukopenia. In total, 13 patients (5.3%) were SFTSV IgM antibody-positive, 48 (19.5%) were IgG-positive. Of the 13 IgM-positive cases, 11 (84.6%) were IgG-positive (9 with titres ≥1:400). Seropositive rates of antibodies were high (8.4% for IgM and 30.1% for IgG) in patients with fever, thrombocytopenia and leukopenia. Furthermore, among IgG-positive cases in this group, 76% (19/25) of patients' IgG antibody titres were ≥1:400. Additionally, 28 of 246 cases were initially diagnosed with suspected SFTS and were then excluded, and 218 patients were never diagnosed with SFTS; the seropositive rates of IgM and IgG in these two groups were 25% and 67.9% and 2.8% and 13.3%, respectively. These rates were 64.3% and 21.4% in 14 sera collected during acute phase of the 28 cases mentioned above. Seropositive rate of SFTSV IgG was only 1.3% in the patient-matched healthy group, and no IgM antibody was detected. A preliminary estimate of 8.3% of SFTS cases were missed in SFTS high endemic provinces. CONCLUSIONS: The actual SFTS incidence was underestimated. Effective measures such as adding a new SFTS case category - "SFTS clinical diagnosis cases" or using serological detection methods during acute phase should be considered to avoid missed diagnoses.

Effects of m-nisoldipine on the activity and mRNA expression of four CYP isozymes in rats.

1. For the first time, a systemic in vivo investigation was employed to evaluate the potential effects of m-nisoldipine on activities of rat cytochrome P450 enzymes (CYP1A2, CYP2C11, CYP2D1 and CYP3A1) by both cocktail probe drugs and the quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR). 2. m-Nisoldipine-treated and blank control groups were respectively administered m-nisoldipine at the dosage of 2.5, 5 and 12.5 mg/kg and CMC-Na solution for 15 days consecutively, then they were given the probe drugs of caffeine, diclofenac, dextromethorphan and midazolam (all probes were 5 mg/kg) by p.o. The blood samples were collected at different times for liquid chromatography coupled with mass spectrometry (LC-MS) analysis. The corresponding pharmacokinetic parameters were applied to evaluate the effects of m-nisoldipine on the four CYP isoforms in vivo. In addition, RT-qPCR was performed to determine the effects of m-nisoldipine on the mRNA expression of CYPs in rat liver. Results indicated that high dose and middle dose of m-nisoldipine showed significant effects on all four CYPs and CYP2C11, respectively. Moreover, for CYP2D1 and CYP1A2, there were no significant effects found at either low or middle dose of m-nisoldipine. 3. This study could provide not only experimental evidence for potential clinical application of m-nisoldipine but also a practical strategy for assessing CYP-mediated drug-drug interactions.

[Amaryllidaceae alkaloids from Lycoris radiata].

Three aporphine-type alkaloids (1-3), three lycorine-type alkaloids (4-6), two crinane type alkaloids (7, 8) and one phenanthridine-type alkaloid (9) were isolated from the chloroform soluble fraction of 70% ethanol extract of the bulbs of Lycoris radiata through various column chromatographies over silica gel, ODS, Sephadex LH-20 and MCI. Their structures were elucidated as (+)-N-methoxylcarbonyl-1,2-methylenedioxyl-isocorydione (1), isocorydione (2), 8-demethyl-dehydrocrebanine (3), (+)-3-hydroxy-anhydrolycorine N-oxide (4), vasconine (5), pancratinine D (6), yemenine A (7), 11-O-acetylhaemanthamine (8), and 5,6-dihydro-5-methyl-2-hydroxyphenanthridine (9) based on their chemical and physicochemical properlies and spectroscopic data. Compound 1 was a new compound and alkaloids 2-9 were isolated and identified from this plant for the first time.