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Yueyan Li, Jialei Zhu, Jing Tang Department of Pharmacy, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, 200090, People’s Republic of ChinaCorrespondence: Jing Tang, Department of Pharmacy, Obstetrics and Gynecology hospital of Fudan University, 128 Shenyang Road, Yangpu District, Shanghai, 200090, People’s Republic of China, Tel/Fax +86 21 33180593, Email 1817@fckyy.org.cnBackground: Traditional medicine is a common treatment option for endometrioid-related symptoms. In the past few decades, Guixiong Xiaoyi formula has been widely used as a traditional medicine for the treatment of endometriosis.Purpose: This study aimed to prepare compound Angelica Ligusticum wallichii granule (CALG) by modern technological methods and to study its pharmacodynamics and mechanisms of treating endometriosis.Methods: The ingredients of CALG were determined by UPLC-Q-TOF/MS. Target prediction of compounds and diseases was performed using databases, and the mechanisms of CALG were predicted by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes and verified by molecular docking. Furthermore, a rat model of endometriosis was established to study the effects of CALG on endometriosis in vivo.Results: CALG with good specificity, durability, and stability was obtained following a detailed preparation process and quality control standard. Using network systems pharmacology, 109 chemical compositions and 104 core targets were identified for the treatment of endometriosis. The composition-target-channel-disease network topology analysis of the top 15 chemical compositions of CALG showed that the beneficial effect of CALG on endometriosis was attributed to phenolic compounds. In addition, CALG treatment reduced the volume of ectopic uterine lesions, promoted apoptosis, inhibited the secretion of inflammatory cytokines, and increased HIF-1 expression in rats with endometriosis.Conclusion: CALG induces apoptosis and inhibits inflammation and is a promising drug for the treatment of endometriosis.Graphical Abstract: Keywords: endometriosis, Angelica Ligusticum wallichii granule, network systems pharmacology, molecular docking, traditional Chinese medicine

Shear Alfvén wave (SAW)/drift Alfvén wave (DAW) fluctuations can be destabilized by energetic particles (EPs) as well as thermal plasma components, which play a key role in the EP energy and momentum transport processes in burning fusion plasmas. The drift Alfvén energetic particle stability (`DAEPS`) code, which is an eigenvalue code using the finite element method (FEM), was developed to analyze Alfvén instabilities excited by EPs. The model equations, consisting of quasi-neutrality condition and Schrödinger-like form of vorticity equation, are derived within the general fishbone-like dispersion relation (GFLDR) theoretical framework, which is widely used to analyze SAW/DAW physics. The mode structure decomposition (MSD) approach and asymptotic matching between the inertial/singular layer and ideal regions are adopted. Therefore, the `DAEPS` code can provide not only frequency and growth/damping rate, but also the parallel mode structure as well as the asymptotic behavior corresponding to the singular layer contribution. Thus, it fully describes fluid and kinetic continuous spectra as well as unstable and damped modes. The model equations have been extended to include general axisymmetric geometry, and to solve for the response of circulating and trapped particles by means of the action angle approach. In this work, we discuss linear dispersion relation and parallel mode structure of drift Alfvén instabilities excited by EPs, computed by the `DAEPS` code with realistic experimental plasma profile and magnetic configuration. Based on this information, we adopt the Dyson-Schrödinger Model (DSM) to further analyze EP energy and momentum flux. We will also briefly discuss how the parallel mode structure of the drift Alfvén instabilities can be used in the DSM to calculate the nonlinear radial envelope evolution and the EP transport.

Yueyan Li,1 Chen Zhang,2 Xuan Huang,1 Jing Tang1 1Department of Pharmacy, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, People’s Republic of China; 2Institute of Reproduction and Development, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, People’s Republic of ChinaCorrespondence: Jing Tang, Department of Pharmacy, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, People’s Republic of China, Tel +86 21 33180593, Fax +86 21 33180593, Email 1817@fckyy.org.cnPurpose: Antimicrobial agents are frequently prescribed during pregnancy. This study aims to explore the association between antimicrobial exposure and pregnancy outcomes.Patients and Methods: A multi-center retrospective cohort study of pregnant patients (n=370) was conducted in 22 tertiary hospitals in China. Adverse pregnancy outcomes and admission of neonate to neonatal intensive care unit (NICU) were considered as outcomes. The effect of antimicrobial exposure on pregnancy outcomes was assessed using a multivariate logistic regression model.Results: Use of first-generation cephalosporins during pregnancy was associated with a significantly higher risk of adverse pregnancy outcomes (odds ratio [OR]: 3.64 [95% confidence interval, CI: 1.43– 9.24], P =  0.007) and admission of neonate to the NICU (OR: 3.41, 95% CI: 1.37– 8.53, P=0.009) compared with use of third-generation cephalosporins, after adjusting for gestational age of exposure to antimicrobial agents, cesarean section, and antimicrobial dose. Similarly, a higher risk of adverse pregnancy outcomes (OR: 14.76, 95% CI: 4.43– 49.11) and neonatal NICU admission (OR: 11.74, 95% CI: 3.59– 38.35) were observed among women with first-generation cephalosporins use compared with mothers with no antimicrobial use.Conclusion: Both first- and third-generation cephalosporins use was associated with an increased risk of adverse pregnancy and neonatal outcomes. In addition, first-generation cephalosporins were associated with an increased risk of those pregnant and neonatal outcomes, when compared with third-generation cephalosporins. We should require to determine the indications and contraindications for use of cephalosporins during pregnancy.Keywords: maternal infections, antimicrobial agents, adverse pregnancy outcomes, NICU, cephalosporin

Recent experiment has uncovered semimetal bismuth (Bi) as an excellent electrical contact to monolayer MoS$_2$ with ultralow contact resistance. The contact physics of the broader semimetal/monolayer-semiconductor family beyond Bi/MoS$_2$, however, remains largely unexplored thus far. Here we perform a comprehensive first-principle density functional theory investigation on the electrical contact properties between six archetypal two-dimensional (2D) transition metal dichalcogenide (TMDC) semiconductors, i.e. MoS$_2$, WS$_2$, MoSe$_2$, WSe$_2$, MoTe$_2$ and WTe$_2$, and two representative types of semimetals, Bi and antimony (Sb). As Bi and Sb work functions energetically aligns well with the TMDC conduction band edge, Ohmic or nearly-Ohmic $n$-type contacts are prevalent. The interlayer distance of semimetal/TMDC contacts are significantly larger than that of the metal/TMDC counterparts, which results in only weak metalization of TMDC upon contact formation. Intriguingly, such weak metalization generates semimetal-induced gap states (MIGS) that extends below the conduction band minimum, thus offering an effective mechanism to reduce or eliminate the $n$-type Schottky barrier height (SBH) while still preserving the electronic structures of 2D TMDC. A modified Schottky-Mott rule that takes into account SMIGS, interface dipole potential, and Fermi level shifting is proposed, which provides an improved agreement with the DFT-simulated SBH. We further show that the tunneling-specific resistivity of Sb/TMDC contacts are generally lower than the Bi counterparts, thus indicating a better charge injection efficiency can be achieved through Sb contacts. Our findings reveal the promising potential of Bi and Sb as excellent companion electrode materials for advancing 2D semiconductor device technology., 12 pages, 7 figures

Background: Thus far, no large-scale clinical study on pregnancy with cancer has been conducted in China. Therefore, the present study aimed to review the oncological characteristics and treatment of pregnancy-associated cancers, and analyze the obstetric and neonatal outcomes to provide evidence-based recommendations for reproductive function preservation, oncological treatment, and obstetric management in pregnancy-associated cancer.Methods: We conducted an observational retrospective cohort study among pregnant patients with cancer in 7 Chinese tertiary A hospitals during 2003–2021. We conducted multiple logistic regression to determine the influence of various factors on preterm birth and small-for-gestational-age infants, log-binomial regression to analyze temporal changes, and chi-square tests to explore the effects of cancer type/treatment.Results: Of 204 women, 17% terminated their pregnancies; 59% received pre-delivery treatment. Every 6 years, the rates of pregnancy termination ([RR]: 0.48, 95% confidence interval [CI]: 0.35–0.67) and iatrogenic preterm births (RR: 0.73, 95% CI: 0.54–0.98) reduced, and that of pre-delivery treatment increased, mainly due to increased rates of surgery (RR: 1.87, 95% CI: 1.31–2.67). Maternal systemic diseases were related to a higher risk of small-for-gestational-age infants ([OR]: 12.02, 95% CI: 1.82–79.43). Chemotherapy with taxanes plus platinum-based agents was related to adverse obstetric outcomes (OR: 1.87, 95% CI: 1.42–2.46, P < 0.05). Thyroid (OR: 0.36, 95% CI: 0.22–0.57) and ovarian cancer (OR: 0.70, 95% CI: 0.50–0.98) were associated with fewer cesarean sections. Thyroid cancer was associated with fetal growth restriction (OR: 5.21, 95% CI: 1.21–22.55).Conclusions: Rates of pregnancy termination in cancer have declined. Taxane plus platinum-based chemotherapy was associated with adverse obstetric outcomes. Cancer type influenced outcomes.

OBJECTIVE The aim of this study was to assess the different impacts on inflammatory biomarkers of ticagrelor or clopidogrel in patients with ST-segment elevation myocardial infarction (STEMI) undergoing emergency percutaneous coronary intervention (PCI). METHODS This was a multicenter, randomized, open-label and parallel- treatment prospective study. Patients were randomized to receive a loading dose of ticagrelor 180 mg as the treatment group (N = 143) or clopidogrel 600 mg as the control group (N = 184). After PCI, the treatment group was treated with ticagrelor 90 mg twice daily while the control group was treated with clopidogrel 75 mg once daily. The levels of inflammation were tested by sCD40 ligand (CD40L), high sensitivity C-reactive protein (hsCRP) and P-selectin in different time points. RESULTS There were 327 patients participated in this study and no difference in baseline characteristics between groups. No difference was found between groups in baseline levels of inflammation with all assays, and no difference in assays of hsCRP and P-selection in all the time points between groups, but sCD40L decreased significantly in ticagrelor group than clopidegrel group (-2492 ± 7407.3 VS 992.21 ± 10478, P = 0.0095) in 1week after PCI. CONCLUSIONS In STEMI patients undergoing PCI who are receiving anti-platelet therapy, ticagrelor is associated with more potent inflammation inhibition of sCD40L compared with clopidogrel in 1week after PCI. It may open a new path for anti-platelet therapy and large studies are needed to support this finding.

To identify small molecules blocking La-RNA interactions by using structural dynamics, molecular biology, and in vivo efficacy experiments.A docking virtual assay on the Chemdiv database was used to screen La binders, and their affinity were measured by surface plasmon resonance (SPR). A novel fluorescence polarization (FP) assay referring to the binding of La protein and 3'UUUOH was established to identify the inhibitors. Their activity on ovarian cancer cell proliferation, apoptosis and cell cycle were evaluated using Cell Counting Kit 8 (CCK8) and flow cytometry assay, respectively. Their in vivo efficacy against ovarian cancer growth were evaluated in a cell line-derived xenograft (CDX) model of A2780 cells.From a total of 20 compounds with high potential binding activity with La protein, two small molecule compounds 4424-1120 and 8017-5932 with relatively stronger inhibition ability on La-RNA interactions were identified. These two compounds shared the same active centers with hydroxyimidazole and hydroxybenzene to interact with La protein through residues ARG57, GLN20 and GLN136. The in vitro assays showed that 4424-1120 and 8017-5932 effectively cause G0/G1 cell cycle arrest, inhibit cell proliferation, reduce cell invasion and promote apoptosis in ovarian cancer cells. In a CDX model on BALB/C Nude mice, we found that the growth rate of the tumor was inhibited by 4424-1120.Our results demonstrated compound 4424-1120 shows good antitumor activity and safety in vitro and in vivo, and it provides a new idea for the discovery of antitumor lead compounds from small drug-like molecules.

Research on the mechanism and new targets of ovarian cancer is of great significance to reduce the high mortality and drug resistance of ovarian cancer. Human La protein has been found to be highly expressed in a variety of malignant tumors and plays a role in tumorigenesis and development through its RNA-binding function. However, its role and mechanism in ovarian cancer are not completely clear. The present study showed that La protein was highly expressed in serum and tissues of patients with ovarian cancer by ELISA and immunohistochemistry, and the high expression of La protein was associated with the increased degree of malignancy and poor prognosis by searching the KM plotter database. Interference of the La gene resulted in a significant decrease in the proliferation, migration, and invasion of ovarian cancer cells with growth block in the G1 phase and increasing apoptosis. By RNA binding protein immunoprecipitation, transcriptome sequencing, and proteomics, 14 downstream target genes were screened. The La protein might affect the protein expression of these 14 genes by binding with the mRNAs. Therefore, it played a role in the pathological process of ovarian cancer.

Primary hyperoxaluria type 1 is a severe inherited disorder that results in recurrent urolithiasis and renal failure. We generated an alanine-glyoxylate aminotransferase ( Agxt) Q84X nonsense mutant rat model that displayed an early onset of hyperoxaluria, spontaneous renal CaOx precipitation, bladder stone, and kidney injuries. Our results suggest an interaction of renal CaOx crystals with the activation of inflammation-, fibrosis-, and necroptosis-related pathways. In all, the AgxtQ84X rat strain has broad applicability in mechanistic studies and the development of innovative therapeutics.

Background Primary hyperoxaluria type 1 (PH1) is a rare genetic disorder caused by endogenous overproduction of hepatic oxalate, leading to hyperoxaluria, recurrent calcium oxalate kidney stones, and end‐stage renal disease. Lactate dehydrogenase (LDH) is an ideal target for diminishing oxalate production as it is responsible for glyoxylate to oxalate conversion in the liver, the last step of oxalate metabolism. Here, we investigated the therapeutic efficacy and potential side effects of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology to ameliorate PH1 via specifically disrupting the hepatic LDH. Methods Pheochromocytoma (PC12) cells were used to assess the efficacy of cleavage of single‐guide RNAs in vitro. PH1 neonatal rats were injected with a single administration of adeno‐associated virus to deliver the CRISPR/Cas9 system that targeted LDH. Three weeks after injection, a liver biopsy was performed to detect LDH expression, liver injury, and liver metabolomics. Urinary oxalate was regularly monitored, and renal calcium oxalate deposition was evaluated after 4 weeks of 0.5% ethylene glycol challenge. After 6 months of treatment, animals were euthanized, and ex‐liver organs were harvested for toxicity analysis. Results The Ldha gene was specifically knocked out in 20% of the liver cells of PH1 rats in the treatment group, leading to a 50% lower LDH expression than that in the control group. Compared to the control groups, urinary oxalate levels were significantly decreased, and renal calcium oxalate precipitation was largely mitigated in the treatment group throughout the entire 6‐month study period. While no CRISPR/Cas9‐associated off‐target edits or hepatotoxicity were detected, we observed mild metabolic changes in the liver tricarboxylic acid (TCA) and glycolysis pathways. Conclusions CRISPR/Cas9‐mediated LDH disruption may represent an applicable new strategy for alleviating PH1 for its long‐lasting effect and low editorial efficiency requirements., CRISPR/Cas9 technology provides novel approaches for inherited monogenic diseases by targeting non‐essential enzymes. Using a rat model, we demonstrate that the AAV‐delivered CRISPR/Cas9 system is an effective and safe platform for knocking down lactate dehydrogenase and alleviating the life‐threatening primary hyperoxaluria type 1.

Reliable pavement surface temperature prediction models are important for decision making in winter road maintenance. Due to the complexity of thermal environment in winter, there is inevitably uncertainty in pavement temperature. Therefore, the prediction results of pavement surface temperature should not be a single value, but a probability distribution. This paper developed a prediction model for evaluating the probability distribution of pavement surface temperature in winter. The model consisted of two modules, namely, a Bayesian Structural Time Series module (BSTS) and a Bayesian Neural Network module (BNN). The BSTS module provided a relatively rough prediction of pavement surface temperature, while the BNN module provided refined prediction based on the results from the BSTS module. The model was validated with data collected from the field test. The impact of model structure, model parameters, and probability threshold on the predictions was analyzed. Results show that most measured pavement temperatures fell into the 95% confidence interval of the predicted values, indicating the prediction was reliable. For the BSTS module, the regression component plays the most important role in the prediction. The seasonal component also has obvious influence on the prediction results. While the influence of local linear trend component on the prediction result is not obvious. For the BNN module, although the slippery index and pavement surface condition do not directly contribute to the heat transfer process, these two factors have correlation with the pavement surface temperature. The value of probability threshold should be properly determined to balance the reliability and computational cost of the prediction model. The model presented in this paper is useful for transportation agencies in winter road maintenance

COVID-19, which broke out in 2019 in China, is caused by SARS-CoV-2. According to the latest WHO real-time statistics, as of 1:19 p.m. BST on April 22, there were 142557268 cumulative new confirmed cases of coronavirus pneumonia and 3033798 cumulative deaths worldwide. Therefore, it is significant to understand the structure and the pathogenic mechanism of SARS-CoV-2, which is helpful to understand the diagnosis and treatment of sars-cov-2. We primarily investigate related sources on Pubmed and related scholar websites. We found some antiviral drugs such as lopinavir, chloroquine, and its derivative hydroxychloroquine, remdesivir, and arbidol have great potential in the treatment of COVID-19. However, some of them have a serious adverse reaction, reminding us to use them with caution.

Under the trend of complex building functions, the paper tries to explore the idea and method of design for gymnasium. Taking the handball hall design of Huyi Campus of Xi’an Institute of Physical Education as an example, the paper puts forward specific design strategies in terms of function allocation, operation mode and image positioning. The purpose is to reveal the inspiration of the design concept of “compound function” for the design and development of gymnasium building.

Fine particulate matter (PM

Adiponectin and miR-133a are key regulators in cardiac hypertrophy. However, whether APN has a potential effect on miR-133a remains unclear. In this study, we aimed to investigate whether APN could regulate miR-133a expression in Angiotensin II (Ang II) induced cardiac hypertrophy in vivo and in vitro. Lentiviral-mediated adiponectin treatment attenuated cardiac hypertrophy induced by Ang II infusion in male wistar rats as determined by reduced cell surface area and mRNA levels of atrial natriuretic peptide (ANF) and brain natriuretic peptide (BNP), also the reduced left ventricular end-diastolic posterior wall thickness (LVPWd) and end-diastolic interventricular septal thickness (IVSd). Meanwhile, APN elevated miR-133a level which was downregulated by Ang II. To further investigate the underlying molecular mechanisms, we treated neonatal rat ventricular myocytes (NRVMs) with recombinant rat APN before Ang II stimulation. Pretreating cells with recombinant APN promoted AMP-activated protein kinase (AMPK) phosphorylation and inhibited ERK activation. By using the inhibitor of AMPK or a lentiviral vector expressing AMPK short hairpin RNA (shRNA) cancelled the positive effect of APN on miR-133a. The ERK inhibitor PD98059 reversed the downregulation of miR-133a induced by Ang II. These results indicated that the AMPK activation and ERK inhibition were responsible for the positive effect of APN on miR-133a. Furthermore, adiponectin receptor 1 (AdipoR1) mRNA expression was inhibited by Ang II stimulation. The positive effects of APN on AMPK activation and miR-133a, and the inhibitory effect on ERK phosphorylation were inhibited in NRVMs transfected with lentiviral AdipoR1shRNA. In addition, APN depressed the elevated expression of connective tissue growth factor (CTGF), a direct target of miR-133a, through the AMPK pathway. Taken together, our data indicated that APN reversed miR-133a levels through AMPK activation, reduced ERK1/2 phosphorylation in cardiomyocytes stimulated with Ang II, revealing a previously undemonstrated and important link between APN and miR-133a.

The Platelet Inhibition and Patient Outcomes (PLATO, Eur J Prev Cardiol 22(6):734–42, 2015) trial shows that, in patients who have an acute coronary syndrome, treatment with ticagrelor as compared with clopidogrel significantly reduced the rate of death, but the reason is still uncertain. Both inflammation and vascular endothelian cell dysfunction play important roles in the pathophysiology of atherosclerotic plaques, but whether ticagrelor has superior anti-inflammatory effect and can improve vascular endothelial cell function to a great extent is unknown. Patients with STEMI who are scheduled to undergo emergency percutaneous coronary intervention (PCI) will be randomly assigned to receive a loading dose of ticagrelor 180 mg as the treatment group or clopidogrel 600 mg as the control group. After PCI, the treatment group will be treated with ticagrelor 90 mg twice daily while the control group will be treated with clopidogrel 75 mg once daily. The vascular endothelial function will be tested by circulating endothelial cells, and levels of inflammation will be tested by CD40 ligand (CD40L), high sensitivity C-reactive protein (hsCRP) and P-selectin. The estimated enrollment sample size will be 350 patients, including 175 in the treatment group and 175 in the control group. This study will compare the influence of ticagrelor and clopidogrel on inflammatory biomarkers and vascular endothelial function firstly for STEMI patients receiving emergency PCI and will provide evidence to identify whether ticagrelor inhibits inflammation and improves vascular endothelial cell function to a greater extent than clopidogrel or not. This trial was registered with Clinicaltrials.gov (identifier: NCT02123004 ) on 20 April 2014.

Heat-sensitive transgene expression systems have been proposed recently for use in gene therapy to enable both spatial and temporal control of the gene activity. The transgene was put under the control of HSP-related promoters and could be turned on by external heat treatment. While the 'heat activation' phenomenon of the HSP-related promoters in vitro had been well documented, the detailed time response and temporal regulation profile in vivo were not fully understood. We reported here the regulation of transgene luciferase expression in vivo in muscles using a custom-built ultrasound-mediated hyperthermia instrument. The effects of different heating parameters and treatment regimens were evaluated. Optimal activation of gene expression was found at 39 degrees C. Significant tissue damage was observed at 41 degrees C and above, which directly correlated with the greatly reduced gene expression. The gene constructs remained stable and silent in muscle cells, and could be turned on at a later time without losing much activity. Repeated activation was also possible, but required heat treatment at a higher temperature to overcome thermotolerance.

Objective: To determine the factors that impact BP control in Chinese elderly (aged 65-79 years) and very elderly (aged ≥80 years) hypertensive patients after 8-week treatment with val/aml SPC in the CSII study. Design and Methods: 3609 elderly and very elderly hypertensive patients uncontrolled by monotherapy with BP ≥140/90 mm Hg (≥130/80 mm Hg for diabetes or CKD) were analyzed. It was a post-hoc analysis of subgroups from the CSII study. All patients received val/aml 80/5 mg SPC for 8 weeks. An additional antihypertensive agent added in patients who were still uncontrolled at the end of 4 weeks. Complications, previous antihypertensive treatment and demographic factors were recorded. Chi-square test, Student’s t-test and logistic regression analysis were conducted. Results: After 8 weeks treatment, 61.5% elderly (n=459) and 63.3% very elderly (n=1811) patients achieved BP control. In the 740 elderly patients, low BP control rates were observed in patients with coronary heart disease (CHD) [P 0.05). Previous antihypertensive treatment (ACEI, ARB, CCB, βblocker and diuretic) had no significant influence on BP control (P>0.05 ) . Among cardiovascular risk factors , while dyslipidemia influenced the BP control rate(P0.05). In very elderly patients, DM led to low BP control (P0.05), but alcohol consumption had a significant impact (P Conclusion: CHD, DM, HF and dyslipidemia negatively influenced BP control in the elderly patients. In the very elderly patients, diabetes and history of alcohol consumption were associated with low BP control rates. The effect of the above risk factors on BP control should be considered during the management of hypertension in the elderly and very elderly Chinese patients with above complications as they would need stricter BP control.

beta-Sitosterol is the most abundant phytosterol lipid in plants. This study demonstrates a method for analyzing beta-sitosterol using a laboratory-built MALDI-TOF mass spectrometer. The matrix compound, solvent system, laser intensity, alkali ion, and analyte concentration for sample preparation were optimized to acquire reproducible and informative mass spectra of the analyte. The best MALDI spectrum of beta-sitosterol was obtained by mixing the analyte solution with an equal volume of 2,4-DHB/iPrOH-H2O ( 9 : 1) solution (60 mM) at 13.5 mJ cm(-2) laser intensity. Under the optimized experimental conditions, beta-sitosterol generated intense peaks consisting of the initial lipids and their oxidation products. The former included mass peaks at m/z 396.6, 397.6, 436.9, and 452.6, which corresponded to the [M - H2O](+), [M + H - H2O](+), [M + Na](+), and [M + K](+) ions of beta-sitosterol, respectively. The latter included mass peaks at m/z 412.7, 413.7, and 431.9, which corresponded to the [M' - H2O](+), [M' + H - H2O](+), and [M' + H](+) ions of mono-oxidized beta-sitosterol, respectively. A quantitative analysis of beta-sitosterol was also conducted using a matrix peak. Calibration curves obtained for beta(-)sitosterol show a good linear range of three orders of magnitude with a regression coefficient square of 0.992. The linear ranges were between 12.1 and 1207.5 mu M. The detection limit was 10 nM. These results demonstrate the potential application of MALDI mass spectrometry for analyzing phytosterols.

Rapid identification of Bacillus spores has attracted great interest in the medical and forensic communities due to the highly pathogenic nature of certain Bacillus species and its potential application in bioweapons. In this study, we evaluated the feasibility of rapid and reliable identification of several Bacillus spp. (B. subtilis, B. cereus, B. megaterium, B. pumilus, and B. sphaericus) using online MALDI-TOF mass spectrometry combined with lipid profiling. The bacterial cultures in liquid medium were extracted with methanol and chloroform, and the extracts were aerosolized into droplets and immediately analyzed with an online MALDI-TOF mass spectrometer. Results showed that phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and diglycosyldiacylglycerol (DGDG) were present in the mass spectra, as alkali metal adducts. The unique lipid profiles obtained from these Bacillus spp. allowed us to differentiate between them. The reliability and reproducibility of the rapid identification method for Bacillus were tested under different experimental conditions. An alternative online chemotaxonomic method for bacterial identification is demonstrated.