Blood metabolites mediate the causal relationship between circulating CX3CL1 levels and prostate cancer: A 2-step Mendelian randomization study.

Chemokines influence the progression of prostate cancer (PCa) through multiple mechanisms. However, the effect of C-X3-C chemokine ligand 1 (CX3CL1) on PCa risk remains controversial. Our study aimed to investigate whether circulating CX3CL1 is causally associated with PCa and to identify metabolites that have mediating effects using the 2-step bidirectional Mendelian randomization (MR) analysis process. Inverse variance weighting (IVW) results were used as the primary observations, while additional sensitivity analyses were conducted. For each standard deviation increase exhibited by the circulating CX3CL1 levels, the risk of PCa was reduced by 0.4% (IVW: OR = 0.996, [95% CI = 0.994-0.998], P < .001), and blood alliin levels increased by 19% (IVW: OR = 1.185, [95% CI = 1.01-1.54], P = .003). For each standard deviation increase in the blood alliin levels, the risk of PCa was reduced by 0.1% (IVW: OR = 0.999, [95% CI = 0.997-0.999], P = .03). Therefore, the protective effect of circulating CX3CL1 on PCa may be mediated by blood alliin levels (mediated proportion = 6.7%). The results supported the notion that high levels of circulating CX3CL1 indicate a lower PCa risk and the idea that the food-derived antioxidant alliin may mediate this association. We emphasize that the use of CX3CL1 as a protective factor against PCa may provide new strategies for PCa prevention and care in the future.

Study on the hypotensive effect and mechanism of hawthorn (Crataegus pinnatifida) fruits and hyperoside in spontaneously hypertensive rats.

Hawthorn fruits have a sweet and sour taste, besides having beneficial therapeutic effects on hyperlipidemia, hypertension, and coronary heart disease, making them widely used in food and clinical medicine. However, their hypotensive effects and potential mechanisms of anti-hypertension still need to be elucidated. This study aims to explore the antihypertensive effect of hawthorn and its monomer hyperoside on spontaneously hypertensive rats through pharmacodynamics, serum metabolomics, and in vivo mechanism studies. After 7 weeks of intervention with hawthorn extract and hyperoside, the blood pressure was significantly reduced. Aortic vascular staining results showed that the injury was significantly improved after intervention with hawthorn extract and hyperoside. According to the serum metabolomics study, the main metabolic pathway regulating blood pressure in hawthorn extract and hyperoside groups was the primary bile acid biosynthesis pathway. Quantitative experiments confirmed that the level of bile acid in the model group was significantly different from that in the normal group, while that in the hawthorn group and the hyperoside group was close to that in the normal group. Based on the prediction of bile acid-hypertension related targets and the literature, nine genes involved in bile acid metabolism and inflammatory pathways were selected for further study. The FXR, TGR5, ET-1, NOS3, Akt1, TNF-α, Ptgs2, ACE2 and Kdr mRNA expression levels in the hawthorn extract and hyperoside groups were significantly different from those in the model groups. In summary, hawthorn extract and hyperoside have a hypotensive effect on spontaneously hypertensive rats through bile acid and inflammation related targets. Hence, hawthorn extract has the potential to become a functional food or an alternative therapy for hypertension.

Full-Color "Off-On" Thermochromic Fluorescent Fibers for Customizable Smart Wearable Displays in Personal Health Monitoring.

Responsive thermochromic fiber materials capable of miniaturization and integrating comfortably and compliantly onto the soft and dynamically deforming human body are promising materials for visualized personal health monitoring. However, their development is hindered by monotonous colors, low-contrast color changes, and poor reversibility. Herein, full-color "off-on" thermochromic fluorescent fibers are prepared based on self-crystallinity phase change and Förster resonance energy transfer for long-term and passive body-temperature monitoring, especially for various personalized customization purposes. The off-on switching luminescence characteristic is derived from the reversible conversion of the dispersion state and fluorescent emission by fluorophores and quencher molecules, which are embedded in the matrix of a phase-change material, during the crystallizing/melting processes. The achievement of full-color fluorescence is attributed to the large modulation range of fluorescence colors according to primary color additive theory. These thermochromic fluorescent fibers exhibit good mechanical properties, fluorescent emission contrast, and reversibility, showing their great potential in flexible smart display devices. Moreover, the response temperature of the thermochromic fibers is controllable by adjusting the phase-change material, enabling body-temperature-triggered luminescence; this property highlights their potential for human body-temperature monitoring and personalized customization. This work presents a new strategy for designing and exploring flexible sensors with higher comprehensive performances.

3D laparoscopic treatment of bladder cancer with pelvic multi-organ invasion: a case report and literature review.

Introduction: Radical cystectomy with dissection of pelvic lymph nodes and urethral diversion is the standard surgical treatment for muscle-invasive non-metastatic bladder cancer. In rare cases where patients with bladder cancer without distant metastasis have pelvic multi-organ invasion, the cancer compresses or invades the ureter and, in severe cases, leads to bilateral upper urinary tract obstruction and renal damage. The treatment recommended by guidelines often cannot improve the patients' clinical symptoms immediately, and patients cannot complete the treatment owing to severe side effects, resulting in poor survival benefits. Case presentation: A 69-year-old woman with facial edema was treated at the First Affiliated Hospital of Jinan University. The serum creatinine and potassium values were 1244 umol/L and 5.86 mmol/L, respectively. Pelvic magnetic resonance and abdominal computed tomography revealed that the bladder tumor had infiltrated the uterus, anterior vaginal wall, rectum, right ureter, right fallopian tube, and right ovary and metastasized to multiple pelvic lymph nodes. Tumor invasion of the right ureter resulted in severe hydronephrosis of the right kidney and loss of function and obstructive symptoms in the left kidney. Four days later, the patient's creatinine level decreased to 98 u mol/L, the general condition significantly improved, and the patient and family members strongly desired surgical treatment of the tumor. Through a comprehensive preoperative discussion, possible intraoperative and postoperative complications were evaluated. Right nephrectomy, right ureterectomy, total pelvic organ resection, extended pelvic lymph node dissection, and bowel and urinary diversion were conducted under 3D laparoscopy-assisted treatment. The patient was followed-up for 1.5 years and showed good tumor control, self-care, and mental status. Conclusion: Minimally invasive surgery is a curative option for patients with bladder cancer with pelvic multi-organ invasion without distant metastasis. Surgeons should strictly control the indications for surgery and warn patients about the occurrence of related post-surgical complications.

Untargeted lipidomics-based study reveals the treatment mechanism of Qingxue Bawei tablets on atherosclerotic in ApoE-/- mice.

Qingxue Bawei (QXBW) tablets, a Mongolian medicine prescription, have proved to possess good lipid-lowering and antihypertensive effects in previous studies. However, the therapeutic effects and potential mechanisms of QXBW tablets on atherosclerosis (AS) have not been well studied yet. This study aimed to investigate the potential liver-protective mechanism of QXBW tablets on AS mice by hepatic lipidomics analysis. After 10 weeks of administration, serum and liver were collected for biochemical, histopathological, and lipid metabolomics analysis to evaluate the efficacy of the QXBW tablets on high-fat diet (HFD) induced mice. The experimental results indicated that QXBW tablets could ameliorate liver injury and inflammatory response in AS mice. Liver lipid data from different groups of mice were collected by UPLC-Q-Orbitrap-MS, and a total of 22 potential biomarkers with significant differences between the model and control groups were identified finally, of which 16 potential biomarkers were back-regulated after the QXBW tablets intervention. These 22 potential differential metabolic markers were mainly involved in glycerolipid metabolism, glycerophospholipid metabolism, and cholesterol ester metabolism pathways. The results of this study showed that serum inflammatory factors, liver function indices, and lipid metabolism disorders were positively alleviated in AS mice after QXBW tablets treatment.

Simultaneous and dynamic measurement of Schisandrol A changes in rat blood and brain and its comparative pharmacokinetic study in control and Parkinson's disease rats by dual-probe in vivo microdialysis.

Schisandrol A (SchA) is the main active ingredient of Schisandra chinensis (Turcz.) Baill., which is a famous traditional Chinese herbal medicine. SchA can penetrate the blood-brain barrier and has a significant neuroprotective effect. A group of multiplexed stable isotope mass tags (MSIMTs, m/z 332, 338, 346, 349, 351, 354, 360, 363, 374 and 377) were synthesized to perform multiplexed stable isotope labeling derivatization (MSILD) of SchA in rat microdialysates and standards. A new magnetic molecularly imprinted polymer was prepared using MSIMT-375-SchA as dummy template. All the 10-plexed derivatives of MSIMTs-SchA can be efficiently and selectively enriched and purified using this adsorbent by magnetic dispersive solid phase extraction (MDSPE) before ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) analysis. It should be pointed out that the MSIMT-346-SchA standard derivative was used as internal standard in the process of MDSPE and UHPLC-MS/MS. On these bases, 9 different rat microdialysate samples can be determined by UHPLC-MS/MS in a single run. The utilization of MSIMTs significantly increased the sensitivity, accuracy, selectivity and analysis throughput. Under the optimized conditions, satisfactory linearity (R2> 0.987), limit of detection (LODs, 0.15-0.26 pg/mL) and lower limit of quantitative (LLOQ, 0.8-2.0 pg/mL) were obtained. Intra- and inter-day precisions were in the range of 2.2% -12.5%, and recoveries 94.2% -106.2%. The matrix effects were very low, and the average derivatization efficiency of 10-plex MSIMTs to SchA was as high as 97.8%. Using the developed dual-probe in vivo microdialysis sampling technique, the proposed analytical method has been applied for comparative pharmacokinetics of SchA in the brain and blood of control and Parkinson's disease (PD) rats.

Affinity ultrafiltration and UPLC-HR-Orbitrap-MS based screening of neuraminidase inhibitors from Angelica pubescens.

Traditional Chinese medicine is a rich source of natural products and has a long history of use because of its remarkable clinical efficacy. In the present study, the chemical constitutes of Angelica pubescens were studied by ultra high performance liquid chromatography and high-resolution Orbitrap mass spectrometry (UPLC-HR-Orbitrap-MS). A total of 78 compounds were identified and the main composition were coumarins and phenolic acids. Then, the neuraminidase was incubated with extract of Angelica pubescens to screen the neuraminidase inhibitors by affinity ultrafiltration methods. As a result, 13 small molecules were discovered to interact with neuraminidase for the first time. In vitro neuraminidase inhibitory activity of the screened compounds and extract of Angelica pubescens was tested, and isochlorogenic acid C, isochlorogenic acid B, osthole, chlorogenic acid, xanthotoxin, phellopterin and imperatorin were proved to have this activity. In addition, molecular docking analysis was conducted to predict the potential docking position. This study may provide a reference for the medical substance basis in Angelica and the clinical usage of this drug.

The cytoplasmic tail substitution increases the assembly efficiency of Ebola virus glycoprotein on the budded virus of Bombyx mori nucleopolyhedrovirus.

Glycoprotein (GP1,2) of the Ebola virus (EBOV) is the key membrane fusion protein, which is a key candidate protein for vaccine preparations. Previously, GP1,2 was expressed by Bombyx mori nucleopolyhedrovirus (BmNPV) expression vector system; however, few GP1,2 was incorporated into budded virus (BV) of BmNPV. To improve the incorporation efficiency of GP1,2 into the virion, the GP1,2 fusion with the cytoplasmic tail of GP64 of BmNPV was expressed in BmN cells by the BmNPV expression system. The BV was purified by ultracentrifugation, and GP1,2 expression in BV was detected by the antibody. The result indicated that a 532% increase in the relative GP1,2 densitometry signal was observed in constructs utilizing the GP64 C-terminal domain; moreover, the substitution of GP1,2 native signal peptide with GP64 signal peptide increased the incorporation efficiency by 34.6% in the relative GP1,2 densitometry signal. We revealed that the application of the cytoplasmic tail of BmNPV GP64 significantly increased the incorporation rate of GP1,2 into the BV envelope. This study lays a foundation for GP1,2 vaccine development.

The Bombyx mori Nucleopolyhedrovirus GP64 Retains the Transmembrane Helix of Signal Peptide to Contribute to Secretion across the Cytomembrane.

Bombyx mori nucleopolyhedrovirus (BmNPV) is the primary pathogen of silkworms that causes severe economic losses in sericulture. GP64 is the key membrane fusion protein that mediates budded virus (BV) fusion with the host cell membrane. Previously, we found that the n-region of the GP64 signal peptide (SP) is required for protein secretion and viral pathogenicity; however, our understanding of BmNPV GP64 remains limited. Here, we first reported that BmNPV GP64 retained its SP in the mature protein and virion in only host cells but did not retain in nonhost cells. Uncleaved SP mediates protein targeting to the cytomembrane or secretion in Bombyx mori cells. The exitance of the n-region extended the transmembrane helix length, which resulted in the cleavage site to be located in the helix structure and thus blocked cleavage from signal peptidase (SPase). Without the n-region, the protein fails to be transported to the cytomembrane, but this failure can be rescued by the cleavage site mutation of SP. Helix-breaking mutations in SP abolished protein targeting to the cytomembrane and secretion. Our results revealed a previously unrecognized mechanism by which SP of membrane fusion not only determines protein localization but also determines viral pathogenicity, which highlights the escape mechanism of SP from the cleavage by SPase. IMPORTANCE BmNPV is the primary pathogen of silkworms, which causes severe economic losses in sericulture. BmNPV and Autographa californica multiple nucleopolyhedrovirus (AcMNPV) are closely related group I alphabaculoviruses, but they exhibit nonoverlapping host specificity. Recent studies suppose that GP64 is a determinant of host range, while knowledge remains limited. In this study, we revealed that BmNPV GP64 retained its SP in host cells but not in nonhost cells, and the SP retention is required for GP64 secretion across the cytomembrane. This is the first report that a type I membrane fusion protein retained its SP in mature proteins and virions. Our results unveil the mechanism by which SP GP64 escapes cleavage and the role of SP in protein targeting. This study will help elucidate an important mechanistic understanding of BmNPV infection and host range specificity.

LC-MS-based lipidomic analysis of liver tissue sample from spontaneously hypertensive rats treated with extract hawthorn fruits.

At present, many experiments provide support for the cardiovascular protective effect of hawthorn (Crataegus oxyacantha) flower, leaf and fruit extracts. The aim of this study was to investigate the intervention mechanism of hawthorn fruit extract on spontaneously hypertensive rats (SHR) and its effect on their lipid metabolic pattern. After SHR was intervened by hawthorn extract (1.08 g/kg/d) for 6 weeks, the blood pressure and liver histopathology of rats were evaluated. An UHPLC-Q Extractive metabolomics approach was used to collect information on rat liver lipid metabolites, combined with multivariate data analysis to identify significantly different substances and potential biomarkers through mass spectrometry and database searches. Histomorphology of the liver was partially restored in the hawthorn-treated group. Hawthorn extract interferes with sphingolipid metabolism, glycerophospholipid metabolism and glycerolipids metabolism, improving partially disturbed metabolic pathways. This study showed that hawthorn could partially restore liver histomorphology and has anti-hypertensive effect by regulating lipid metabolism.

Efficacy and Adverse Reactions of Arthroscopic Half-Moon Plate Invasive Surgery in Patients with Acute Knee Pain (AKP): Systematic Review and Meta-Analysis.

Objective: To evaluate the efficacy and adverse reactions of arthroscopic half-moon plate invasive surgery (DEB) in patients with knee pain (AKP) using meta-analysis techniques. Methods: The computer retrieves from the English databases PubMed, EMBASE, Cochrane Library, and Web of Science and the Chinese databases China Knowledge Network, Wanfang Database, VIP Database, and China Biomedical Literature Database to collect information about DEB therapeutic AKP randomized controlled trial. Develop criteria for documentation inclusion and exclusion, evaluate the quality and bias risk of literature, and compare differences in efficacy and adverse responses before and after DEB treatment and other conservative treatments. Results: A total of 14 randomized controlled trials and 1464 AKP patients were included in the study, with follow-up duration of 1-12 months. The total knee scores for DEB at 1, 3, 6, and 12 months after treatment were significantly better than baseline levels (1 month: WMD = 34.56, P = 0.02; 3 months: WMD = 27.73, P = 0.0001; 6 months: SMD = 2.38, P = 0.0001; 12 months: SMD = 1.69, P = 0.001). At 6 months of follow-up, DEB improved knee function better than HA (SMD = 0.47, P = 0.003), and during follow-up for 12 months, DEB relieved knee pain (SMD = 0.55, P = 0.0007) and improved knee function (SMD = 0.88, P = 0.0001) which is significantly better than HA. DEB was less effective at improving knee function in 1, 3, and 12 months than DEB joint HA injections (1 month P = 004; 3 months P = 0.01; 12 months P = 0.03). At 6 and 12 months of follow-up, DEB was better at pain relief and improved function than ozone (P > 0.05). DEB and glucocorticosteroids have similar effects in pain relief and improved function at various follow-up times. In terms of adverse reactions, DEB does not increase the risk of adverse events compared to HA (OR = 0.96, P = 0.85). Conclusion: Compared to HA and ozone, DEB is a more effective treatment for AKP joints, while DEB is combined with HA. The clinical efficacy of injection therapy AKP is better than that of DEB alone.

Effects of bariatric surgery on testosterone level and sexual function in men with obesity: A retrospective study.

Introduction: Bariatric surgeries induce well-documented weight loss and resolve obesity comorbidities. Sexual function is one of the aspects of life quality and may benefit from surgery. Few studies have revealed the impact of bariatric surgeries on sexual function in Chinese men with obesity. Methods: This is a retrospective cohort study of patients undergoing bariatric surgery [laparoscopic sleeve gastrectomy (LSG) or laparoscopic Roux-en-Y gastric bypass (LRYGB)]. Data were collected between September 2017 and February 2022. The International Index of Erectile Function (IIEF) questionnaire was used to evaluate erectile function, intercourse satisfaction, orgasmic function, sexual desire, and overall satisfaction. Sex hormones and other blood tests were evaluated before and at least 1 year after the surgery. Results: Fifty-nine Chinese male patients completed the IIEF questionnaire. The multivariate logistic regression analysis revealed that body mass index (BMI) was the single independent risk factor of the severity of erectile dysfunction (ED). Preoperative testosterone levels had negative correlations with BMI and waist circumference. Thirty-seven patients completed the postoperative questionnaire with a mean follow-up of 23.2 months. Conclusion: BMI and waist circumference were negatively correlated with testosterone levels. BMI was an independent risk factor for the severity of ED. LSG and LRYGB led to positive and sustained improvement in sexual function of men with obesity. The two procedures had a comparable effect, more subjects being needed. Sex hormone levels also could be reversible. However, more weight loss did not predict a positive change in sexual function. A greater BMI loss might predict a greater increase in testosterone.

Data-Driven Discrimination, Perceived Fairness, and Consumer Trust-The Perspective of Consumer Attribution.

With the development of consumer-centric data collection, storage, and analysis technologies, there is growing popularity for firms to use the behavioral data of individual consumers to implement data-driven discrimination strategies. Different from traditional price discrimination, such data-driven discrimination can take more diverse forms and often discriminates particularly against firms' established customers whom firms know the best. Despite the widespread attention from both the academia and the public, little research examines how consumers react to such discrimination enabled by big data. Based on attribution theory, this paper examines how different ways of consumer attribution of data-driven discrimination influence perceived fairness and consumer trust toward the firm. Specifically, we hypothesize that controllability by consumers and locus of causality of data-driven discrimination interactively influence perceived fairness, which further affects consumer trust. We conduct two experiments to test the hypotheses. Study 1 uses a 2(controllability: high vs. low)×2(locus of causality: internal vs. external) between-subjects design. The results show a significant interaction between controllability and locus of causality on consumer trust. When consumers attribute data-driven discrimination to themselves (internal attribution), consumer trust is significantly lower in low-controllable situations than that in high-controllable situations. When consumers attribute the discrimination to the firm (external attribution), however, the impact of controllability on consumer trust is nonsignificant. Moreover, we show that perceived fairness plays a mediating role in the interaction effect of controllability and locus of causality on consumer trust. Study 2 uses a similar design to replicate the findings of Study 1 and further examines the moderating role of consumer self-concept clarity. The results show that the findings of study 1 apply only to consumers with low self-concept clarity. For consumers with high self-concept clarity, regardless of the locus of causality (internal or external), consumer trust is significantly higher in high-controllable situations than that in low-controllable situations. Finally, we discuss the theoretical and managerial implications and conclude the paper by pointing out future research directions.

Simultaneous Determination and Comprehensive Evaluation of Mineral Elements in Yellow Croaker Ear-Stone on Sale in China.

Yellow Croaker Ear-stone or Yunaoshi, is actually two kinds of fish otolith in China and has received increased attention in recent years as important folk medicine. For better understanding of this crude drug, a chaotic market circulation status investigation was carried out and seventeen samples with different varieties or producing areas were collected. In this study, pharmacodynamic components of nineteen varieties mineral elements of the seventeen samples were simultaneously determined by Inductively coupled plasma mass spectrometry (ICP-MS) method. The detected elements were categorized into the beneficial (Na, Mg, Ca, K, Fe, Mn, Zn, Sr, B) and unbeneficial elements (Cu, As, Cd, Hg, Al, Pb, Co, Ba, Cr and Ni) kinds and their concentrations were quantified. Then the principal component analysis (PCA) and hierarchical clustering analysis (HCA) were further applied to launch an exploratory analysis for Yunaoshi samples. The results showed that samples 1-3, 15-8, 15-3 ranked the top three from the perspective of beneficial elements and samples 1-3, 1-4, 15-2 ranked the top three based on the unbeneficial elements sides. Combined with HCA results, all samples can be used as the substitutes for Yunaoshi except for samples 1-3, 1-4 and 15-2 only judging from the perspective of mineral elements concentrations. In conclusion, simultaneous determination of mineral elements accompanied with PCA and HCA can not only provide pharmacogenetic reference for the medicinal material of Yunaoshi, but also establish a feasibility for exploring new crude resources or substitutes to this medicine.

Derivatization-based magnetic dummy molecularly imprinted polymers integrated with 4-plex stable isotope labeling derivatization strategy for specific and rapid determination of L-hydroxyproline in human serum.

The specific determination of L-hydroxyproline (Hyp) can serve as a potential indicator for early clinical diagnosis of liver fibrosis. In this work, an integrated strategy based on 4-plex stable isotope labeling derivatization combined with dummy magnetic molecularly imprinted polymers (QSILD-DMMIPs) was developed for specific extraction and rapid determination of Hyp in human serum by ultra high performance liquid chromatography tandem mass spectrometry. A new series of QSILD reagents d0/d1/d2/d3-6-N-methyl-rhodamine 6G-N-hydroxysuccinimidyl formate (d0/d1/d2/d3-MRSF) were designed, synthesized and applied for the high-throughput labeling of Hyp in serum samples. The structural analogue derivative of Hyp with 6-N-ethyl-rhodamine 6G-N-hydroxysuccinimidyl formate (ERSF-Hyp) was synthesized and used as a novel dummy template to prepare DMMIPs. The DMMIPs were well characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), fourier transform infrared spectroscopy (FTIR), Brunner Emmet Teller (BET) measurements, thermogravimetric analysis (TGA), X-ray diffraction (XRD), zeta potential and adsorption experiments. All d0/d1/d2/d3-MRSF-Hyp derivatives were conveniently and specifically adsorbed by DMMIPs in magnetic dispersive solid phase extraction procedure before injection. Method validation results including linearity (0.2-100 ng mL-1), limits of detection and quantitation (0.05 and 0.2 ng mL-1), accuracy, precision, stability, matrix effect and derivatization efficiency were satisfactory. The analytical performances benefited from efficient integration of QSILD and specific DMMIPs extraction. The proposed strategy was successfully applied for Hyp determination in human serum of liver fibrosis patients and healthy controls, which was of great significance to early diagnosis.

9-Plex ultra high performance liquid chromatography tandem mass spectrometry determination of free hydroxyl polycyclic aromatic hydrocarbons in human plasma and urine.

Hydroxyl-polycyclic aromatic hydrocarbons (OH-PAHs) are biomarkers for assessing the exposure levels of polycyclic aromatic hydrocarbons (PAHs). A series of stable isotope mass tags (SIMT-332/338/346/349/351/354/360/363/374/377) were firstly designed and synthesized to perform multiplexed stable isotope labeling derivatization (MSILD) of OH-PAHs in human plasma and urine. Their derivatives were enriched and purified by magnetic dispersive solid phase extraction (MDSPE) using prepared Fe3O4/GO and then determined by ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) in multiple reaction monitoring mode. 9-Plexed MSILD reagents were prepared using pipemidic acid as core structure with different isotope mass tags, in which carbonyl chloride group was used to label OH-PAHs. The SIMT-346 labeled OH-PAHs standards were used as internal standards, which can greatly increase the quantitative accuracy. 9-Plex labeled nine different real samples can be quantified by UHPLC-MS/MS in a single run. Under optimized MSILD-MDSPE conditions, good linearities of seven OH-PAHs were obtained with satisfactory coefficient of determination R2 > 0.991. Limits of detection (LODs) of seven OH-PAHs were from 0.1 to 0.5 pg/mL, and limits of quantitation (LOQs) ranged from 0.5 to 2.0 pg/mL. The intra- and inter-day precisions ranged in 2.3-12.4% with accuracies in the range of 91.7-108.4%. Acceptable results of matrix effect (89.7-105.7%) and derivatization efficiency (> 96.4%) were obtained. In short, the developed method has been proved to be high-throughput, sensitive, accurate and easy-handling. This method was applied for the measurement of seven free OH-PAHs in human urine and plasma, and expected to provide technical support for the evaluation of PAHs exposure levels in humans.

Efficient Expression and Processing of Ebola Virus Glycoprotein Induces Morphological Changes in BmN Cells but Cannot Rescue Deficiency of Bombyx Mori Nucleopolyhedrovirus GP64.

Ebola virus (EBOV) disease outbreaks have resulted in many fatalities, yet no licensed vaccines are available to prevent infection. Recombinant glycoprotein (GP) production may contribute to finding a cure for Ebola virus disease, which is the key candidate protein for vaccine preparation. To explore GP1,2 expression in BmN cells, EBOV-GP1,2 with its native signal peptide or the GP64 signal peptide was cloned and transferred into a normal or gp64 null Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid via transposition. The infectivity of the recombinant bacmids was investigated after transfection, expression and localization of EBOV-GP were investigated, and cell morphological changes were analyzed by TEM. The GP64 signal peptide, but not the GP1,2 native signal peptide, caused GP1,2 localization to the cell membrane, and the differentially localized GP1,2 proteins were cleaved into GP1 and GP2 fragments in BmN cells. GP1,2 expression resulted in dramatic morphological changes in BmN cells in the early stage of infection. However, GP1,2 expression did not rescue GP64 deficiency in BmNPV infection. This study provides a better understanding of GP expression and processing in BmN cells, which may lay a foundation for EBOV-GP expression using the BmNPV baculovirus expression system.

Integration of stable isotope labeling derivatization and magnetic dispersive solid phase extraction for measurement of neurosteroids by in vivo microdialysis and UHPLC-MS/MS.

In this work, a novel strategy of stable isotope labeling derivatization (SILD) combined with magnetic dispersive solid-phase extraction (MDSPE), has been proposed for simultaneous monitoring of neurosteroids changes linked to Parkinson's disease (PD) by in vivo microdialysis. The developed method was based on ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) detection using multiple-reaction monitoring (MRM) mode. In this study, a new pair of stable isotope labeling reagents d0-/d3-3-N-methyl-2'-carboxyl Rhodamine 6G (d0-/d3-MCR6G), were designed and synthesized for derivatizing neurosteroids in rat blood microdialysates and neurosteroid standards, respectively. d3-MCR6G labeled neurosteroids standards were used as internal standard for the following pretreatment and UHPLC-MS/MS quantification to minimize the deviations caused by complex matrix and ion suppression effects in mass spectrometry analysis. Under the optimized derivatization and extraction conditions, good linearities of eight neurosteroids were obtained with correlation coefficients R2 values > 0.98. The limits of detection (LODs) and quantitation (LOQs) ranged from 0.06 to 0.12 pg/mL and 0.30-0.40 pg/mL, respectively. Taken together, the established method exhibited high sensitivity and selectivity, excellent accuracy, convenience and high efficiency. It was applied for the simultaneous and dynamic measurement of multiple neurosteroids in normal and PD rat blood microdialysates. This method would be expected to be potentially useful for the monitoring and drug treatment of PD and related neurological disorders in the future.

Alcohol dehydrogenase of Candida albicans triggers differentiation of THP-1 cells into macrophages.

Candida albicans proteins located on the cell wall and in the cytoplasm have gained great attention because they are not only involved in cellular metabolism and the maintenance of integrity but also interact with host immune systems. Previous research has reported that enolase from C. albicans exhibits high immunogenicity and effectively protects mice against disseminated candidiasis. In this study, alcohol dehydrogenase (ADH) of C. albicans was cloned and purified for the first time, and this study focused on evaluating its effects on the differentiation of the human monocytic cell line THP-1. The morphological features of THP-1 cells exposed to ADH were similar to those of phorbol-12-myristate acetate-differentiated (PMA-differentiated) macrophages. Functionally, ADH enhanced the adhesion, phagocytosis, and killing capacities of THP-1 cells. A flow cytometric assay demonstrated that ADH-induced THP-1 cells significantly increased CD86 and CD11b expression. The production of IL-1ß, IL-6, and TNF-α by cells increased in the presence of ADH. As expected, after pretreatment with a MEK inhibitor (U0126), ADH-induced THP-1 cells exhibited unaltered morphological features, eliminated ERK1/2 phosphorylation, prevented CD86/CD11b upregulation and inhibited pro-inflammatory cytokine increase. Collectively, these results suggest that ADH enables THP-1 cells to differentiate into macrophages via the ERK pathway, and it may play an important role in the immune response against fungal invasion.

Stable isotope labeling derivatization coupled with magnetic dispersive solid phase extraction for the determination of hydroxyl-containing cholesterol and metabolites by in vivo microdialysis and ultra-high performance liquid chromatography tandem mass spectrometry.

Hydroxyl-containing cholesterol and metabolites (HCMs) are potential biomarkers for Alzheimer's disease. Therefore, quantitative analysis of HCMs can serve as an indicator of clinical diagnosis and drug treatment. In this work, we developed an accurate, sensitive and rapid method for the determination of HCMs in rat blood microdialysates by microwave-assisted stable isotope labeling derivatization (MA-SILD) magnetic dispersive solid phase extraction (MDSPE) coupled with ultra-high performance liquid chromatography electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) in the multiple reaction monitoring mode (MRM). In this respect, a pair of new SILD reagents, d0-/d3-3-N-methyl-2'-carboxyl Rhodamine 6G (d0-/d3-MCR6G), were designed, synthesized and used to label HCMs. Rhodamine 6G with permanently positive charge was introduced into HCMs, improving ionization efficiency and enhancing detection sensitivity. In addition, the d3-MCR6G labeled standards were served as internal standards, reducing the matrix effect and guaranteeing accurate quantification. Various factors affecting MA-SILD and MDSPE were optimized. Furthermore, good linearity was obtained with R2 > 0.994 over the concentration range of 2-3000 pg/mL. The limits of detection (LODs) and quantitation (LOQs) were in the range of 0.24-0.31 and 1.5-1.8 pg/mL, respectively. Acceptable precision (1.6-12.6%), accuracy (87.9-105.2%), matrix effect (85.4-111.2%) and derivatization efficiency (>98%) were achieved. On the whole, this method was validated and applied to accurate, sensitive and simple quantitation of HCMs in rat blood microdialysates. This work would provide some technical support in the diagnosis and treatment of AD or other neurological diseases.