Fecal calprotectin is a novel biomarker to predict the clinical outcomes of patients with ruptured intracranial aneurysm.

BACKGROUND: Intracranial aneurysm (IA) is a common cerebrovascular disease and the leading cause of spontaneous subarachnoid hemorrhage. Recent evidence suggests that gut microbiota is involved in the pathophysiological process of IA through the gut-brain axis. However, the role of gut inflammation in the development of IA has yet to be clarified. Our study aimed to investigate whether fecal calprotectin (FC) level, a sensitive marker of gut inflammation, is correlated with the development of IA and the prognosis of patients with ruptured IA (RIA). METHODS: 182 patients were collected from January 2022 to January 2023, including 151 patients with IA and 31 healthy individuals. 151 IA patients included 109 patients with unruptured IA (UIA) and 42 patients with RIA. The FC level was measured by enzyme-linked immunosorbent assay. Other detailed information was obtained from an electronic medical record system. RESULTS: Compared with healthy controls, the FC levels in patients with IA were increased (P < 0.0001). Patients with RIA had significantly higher FC levels than UIA patients (P < 0.0001). Moreover, the FC level in RIA patients with unfavorable outcomes was higher than in RIA patients with favorable outcomes. Logistic regression analysis showed that the elevated FC level was an independent risk factor for a 3-month poor prognosis in patients with RIA (OR=1.005, 95% CI = 1.000 -1.009, P = 0.044). CONCLUSION: Fecal calprotectin level is significantly elevated in IA patients, especially those with RIA. FC is a novel biomarker of 3-month poor outcomes in RIA patients.

[Microorganisms in the urethral secretions of chronic prostatitis patients: A genomic analysis].

OBJECTIVE: To investigate the differences in the microbial floras in the urethral secretions of the patients with chronic prostatitis to provide some reliable pathogenic evidence for the diagnosis and treatment of the disease. METHODS: Using high-throughput second-generation sequencing technology, we detected the microorganisms in the urethral secretions from 33 chronic prostatitis patients and 30 normal healthy males. We analyzed the significant differences in the microbial flora between the two groups via the rank-sum test and performed data processing with the bioinformatics software, P < 0.05 considered as with statistically significant difference. RESULTS: Statistically significant differences were observed in 17 kinds of bacteria from the urethral secretions between the normal healthy controls and chronic prostatitis patients. LEfSe analysis showed that the microorganisms with most significant abundance difference in the urethral secretions of the chronic prostatitis patients included micrococcaceae, coriobacteriaceae, coriobacteriales, coriobacteriia, weeksellaceae, comamonadaceae, enterobacteriaceae, enterobacteriales, xanthomonadaceae, and xanthomonadales. Principal component analysis (PCA) revealed significant difference in the microbial composition between the two groups. CONCLUSIONS: There is a certain correlation between chronic prostatitis and changes in the composition of urethral microbial floras. Chronic prostatitis may result from concerted action of multiple microbes rather than a single one.

Determination of L-hydroxyproline using hydrophilic interaction chromatography coupled to tandem mass spectrometry with lyophilized concentrated extraction in milk and dairy products.

In this study, a screening and confirmation method for the determination of l-hydroxyproline (Hyp) as a target compound in milk and dairy products using high-performance liquid chromatography with tandem mass spectrometry was developed. The samples were lyophilized after acidic hydrolysis, followed by cleanup with graphitized carbon black to remove pigments. Hyp was separated by a hydrophilic interaction chromatographic column and analyzed by high-performance liquid chromatography with tandem mass spectrometry working with multiple reaction monitoring mode using an electrospray ionization interface in a positive-ion mode. Average recoveries in spiked milk and dairy products ranged from 68.0 to 101.1% with relative standard deviations between 2.0 and 11.7% (n = 7). A reagent-matched standard calibration curve was used for quantification of Hyp, with linear correlation coefficient (R(2)) > 0.99 in the concentration range of 0.1-100 µg/mL. The LOQs were from 0.25 to 5 mg/kg, which were usually sufficient to verify the Hyp in samples. The confirmation concentration of Hyp ranged from 10 to 50 mg/kg.

Relationship between tyrosine phosphorylation and protein expression of insulin receptor and insulin resistance in gestational diabetes mellitus.

The relationship between tyrosine phosphorylation (TP) and protein expression of insulin receptor (InsR) and insulin resistance (IR) in patients with gestational diabetes mellitus (GDM) was investigated. The InsR expression and TP in skeleton muscle tissue were determined by Western blotting and immunoprecipitation in women with GDM (GDM group, n=22), normal pregnant women (normal pregnancy group, n=22) and normal non-pregnant women (normal non-pregnant group, n=13). Fasting plasma glucose (FPG) and fasting insulin (FINS) were measured by oxidase assay and immunoradioassay. The results showed that the levels of FPG (5.61±0.78 mmol/L), FINS (15.42±5.13 mU/L) and Homeostasis model assessment-IR (HOMA-IR) (1.21±0.52) in GDM group were significantly higher than those in normal pregnancy group (4.43±0.46 mmol/L, 10.56±3.07 mU/L and 0.80±0.31 respectively) (P<0.01). The levels of FINS and HOMA-IR in normal pregnancy group were significantly higher than those in normal non-pregnant group (7.56±2.31 mU/L and 0.47±0.26 respectively) (P<0.01). There was no significant difference in the InsR expression level among the three groups (P>0.05). TP of InsR with insulin stimulation was significantly decreased in GDM group (0.20±0.05) as compared with normal pregnancy group (0.26±0.06) (P<0.01). TP of InsR with insulin stimulation in normal pregnancy group was lower than that in normal non-pregnant group (0.31±0.06) (P<0.01). TP of InsR with insulin stimulation was negatively related with HOMA-IR in GDM group (r=-0.525, P<0.01). There was no correlation between the protein expression of InsR and HOMA-IR in GDM group (r=-0.236, P>0.05). It was suggested that there is no significant correlation between the protein expression of InsR in skeletal muscle and IR in GDM, but changes in TP of InsR are associated with IR in GDM.

Van der Waals Schottky Junction Photodetector with Ultrahigh Rectifying Ratio and Switchable Photocurrent Generation.

Metal-semiconductor junctions play an important role in the development of electronic and optoelectronic devices. A Schottky junction photodetector based on two-dimensional (2D) materials is promising for self-powered photodetection with fast response speed and large signal-to-noise ratio. However, it usually suffers from an uncontrolled Schottky barrier due to the Fermi level pinning effect arising from the interface states. In this work, all-2D Schottky junctions with near-ideal Fermi level depinning are realized, attributed to the high-quality interface between 2D semimetals and semiconductors. We further demonstrate asymmetric diodes based on multilayer graphene/MoS2/PtSe2 with a current rectification ratio exceeding 105 and an ideality factor of 1.2. Scanning photocurrent mapping shows that the photocurrent generation mechanism in the heterostructure switches from photovoltaic effect to photogating effect at varying drain biases, indicating both energy conversion and optical sensing are realized in a single device. In the photovoltaic mode, the photodetector is self-powered with a response time smaller than 100 µs under the illumination of a 405 nm laser. In the photogating mode, the photodetector exhibits a high responsivity up to 460 A/W originating from a high photogain. Finally, the photodetector is employed for single-pixel imaging, demonstrating its high-contrast photodetection ability. This work provides insight into the development of high-performance self-powered photodetectors based on 2D Schottky junctions.

[Design and development of trace Cr(VI) sensor].

Hexavalent chromium detection in medicine capsules is generally analyzed in the laboratory, it is difficult to meet the demand for field detection, and to address this problem, a sensor which can be used for on-site detection of trace amounts of hexavalent chromium was designed. It mainly includes chemically sensitive materials, optical sensing module and signal processing module, the chemical sensitive materials is to achieve the conversion of the hexavalent chromium concentration signal, the optical sensing module is to complete a stable output of the laser light source, and the signal processing module is to complete a photoelectric conversion of the weak fluorescence signal, signal amplification, and data processing and displaying. With using the indigenously developed photoelectric acquisition, conversion and signal processing system to complete the rapid detection of trace amounts of hexavalent chromium, so the miniaturization of testing instruments and on-site detection were achieved. Experimental results show that: the sensor detection results have a good linear relationship when the hexavalent chromium concentration is 10-500 microg x L(-1), the linear equation is Y = 1.542 47 x X-2.353 47, and the linearity is 0.998 62, the detection limit reaches 10 microg x L(-1), the sensor response time is about 90 seconds, 5 capsule samples were selected to do the contrast detection, and the results show that the sensor quantitative detection data is reliable, which meets trace hexavalent low cost, fast and field detection demands.

Development of an enzyme-linked immunosorbent assay for determination of the furaltadone etabolite, 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) in animal tissues.

OBJECTIVE: To determine 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) residues released from protein bound AMOZ in animal tissues. METHODS: Polyclonal and monoclonal antibodies were produced in this study. A rapid, sensitive, and specific competitive direct enzyme-linked immunosorbent assay (cdELISA) was developed. RESULTS: Rabbit polyclonal antibodies were used in the optimized cdELISA method, and exhibited negligible cross-reactivity with other compounds structurally related to AMOZ. The IC(50) of the polyclonal antibody was 0.16 ng/mL. The method limit of detection in four different types of animal and fish tissues was less than 0.06 µg/kg. Recoveries ranged from 80% to 120% for fortified samples with the coefficient of variation values less than 15%. The results of the cdELISA method were in good agreement with the results from an established liquid chromatography-tandem mass spectrometry confirmatory method used for AMOZ residues. CONCLUSION: The cdELISA method developed in the present study is a convenient practical tool for screening large numbers of animal and fish tissue samples for the the detection of released protein bound AMOZ residues.

Genome-wide identification, classification and analysis of heat shock transcription factor family in maize.

BACKGROUND: Heat shock response in eukaryotes is transcriptionally regulated by conserved heat shock transcription factors (Hsfs). Hsf genes are represented by a large multigene family in plants and investigation of the Hsf gene family will serve to elucidate the mechanisms by which plants respond to stress. In recent years, reports of genome-wide structural and evolutionary analysis of the entire Hsf gene family have been generated in two model plant systems, Arabidopsis and rice. Maize, an important cereal crop, has represented a model plant for genetics and evolutionary research. Although some Hsf genes have been characterized in maize, analysis of the entire Hsf gene family were not completed following Maize (B73) Genome Sequencing Project. RESULTS: A genome-wide analysis was carried out in the present study to identify all Hsfs maize genes. Due to the availability of complete maize genome sequences, 25 nonredundant Hsf genes, named ZmHsfs were identified. Chromosomal location, protein domain and motif organization of ZmHsfs were analyzed in maize genome. The phylogenetic relationships, gene duplications and expression profiles of ZmHsf genes were also presented in this study. Twenty-five ZmHsfs were classified into three major classes (class A, B, and C) according to their structural characteristics and phylogenetic comparisons, and class A was further subdivided into 10 subclasses. Moreover, phylogenetic analysis indicated that the orthologs from the three species (maize, Arabidopsis and rice) were distributed in all three classes, it also revealed diverse Hsf gene family expression patterns in classes and subclasses. Chromosomal/segmental duplications played a key role in Hsf gene family expansion in maize by investigation of gene duplication events. Furthermore, the transcripts of 25 ZmHsf genes were detected in the leaves by heat shock using quantitative real-time PCR. The result demonstrated that ZmHsf genes exhibit different expression levels in heat stress treatment. CONCLUSIONS: Overall, data obtained from our investigation contributes to a better understanding of the complexity of the maize Hsf gene family and provides the first step towards directing future experimentation designed to perform systematic analysis of the functions of the Hsf gene family.

[Fluorescence resonance energy transfer between gentamycin and water-soluble CdTe QDs].

The water-soluble CdTe quantum dots (QDs) were prepared by using mercaptopropionic acid (MPA) as stabilizer in the aqueous system. Fluorescence resonance energy transfer (FRET) system was constructed between gentamycin (acceptor) and water-soluble CdTe QDs (donor). The maximal emission wavelength was 690 nm, and the line width of the fluorescence spectrum was very narrow (with the full width at half-maximum about 10 nm) and symmetric. The transfer of resonance energy from the CdTe QDs to gentamycin (GT) resulted in the fluorescence quenching of the QDs, corresponding to the increase in the concentration of GT. Several factors that impacted the fluorescence spectra of the FRET system, such as the excitation wavelength (305-425 nm), pH(5.0-11.0), ions (0-0.1 mmol x L(-1) PBS; 0-0.5 mmol x L(-1) NaCl), time (1-120 min), temperature (5-50 degrees C), and concentration of GT (2-80 mg x L(-1)), were investigated and refined. The linear ranges of GT concentration were 2-20 mg x L(-1), r = 0.986 7. Fourier transform infrared spectroscopy (FTIR) and high-performance liquid chromatography (HPLC) were used for confirming the chemical construction and relative specificity, respectively. The results indicated that sulfur and oxygen atoms in MPA molecules took part in coordination with rich Cd2+ on the surface of the nanoparticles. Meanwhile the results also demonstrated that the hydrogen bond between carboxyl of mercaptopropionic acid on the surface of quantum dots and amidocyanogen of GT mainly contributes to combining CdTe with GT. The combination ratio between GT and CdTe QDs is 0.35 to 1.0 according to HPLC. GT as an enhancement has first been applied to the determination of the bovine serum albumin (BSA) labeled with CdTe QDs, and the fluorescence intensity of the labeled BSA with GT is 6 times higher than the control. The proposed method might offer an attractive potential for use in future, because it is sensitive and rapid.

Induced Pluripotent Stem Cells in Huntington's Disease: Disease Modeling and the Potential for Cell-Based Therapy.

Huntington's disease (HD) is an incurable neurodegenerative disorder that is characterized by motor dysfunction, cognitive impairment, and behavioral abnormalities. It is an autosomal dominant disorder caused by a CAG repeat expansion in the huntingtin gene, resulting in progressive neuronal loss predominately in the striatum and cortex. Despite the discovery of the causative gene in 1993, the exact mechanisms underlying HD pathogenesis have yet to be elucidated. Treatments that slow or halt the disease process are currently unavailable. Recent advances in induced pluripotent stem cell (iPSC) technologies have transformed our ability to study disease in human neural cells. Here, we firstly review the progress made to model HD in vitro using patient-derived iPSCs, which reveal unique insights into illuminating molecular mechanisms and provide a novel human cell-based platform for drug discovery. We then highlight the promises and challenges for pluripotent stem cells that might be used as a therapeutic source for cell replacement therapy of the lost neurons in HD brains.

Preparation and characterization of an immunoaffinity column for the selective extraction of aflatoxin B1 in 13 kinds of foodstuffs.

A rapid and reliable immunoaffinity column (IAC) clean-up based ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of aflatoxin B1 (AFB1) in cereals, peanuts, vegetable oils and Chinese traditional food products like sufu and lobster sauce. The immunoaffinity column of AFB1 (AFB1-IAC) was prepared by coupling CNBr-activated Sepharose-4B with the anti-AFB1 monoclonal antibody. The column capacity of IAC was over 260ng/mL gel. Samples were extracted with methanol-water (60:40, v/v) and the extracts were then purified on an AFB1-IAC before UPLC-MS/MS analysis. The average recoveries of AFB1 in spiked samples at levels of 1.0, 5.0 and 10.0µg/kg ranged from 72% to 98%, with the relative standard deviations of 1.2-9.3% (n=6). The limits of qualification ranged from 0.07 to 0.23µg/kg, which were below the MRLs of AFB1 in the matrices evaluated. In this work, the developed method was suitable for the determination of trace AFB1 residues in 13 kinds of foodstuffs.

Ultra-performance liquid chromatography-tandem mass spectrometry determination and depletion profile of flunixin residues in tissues after single oral administration in rabbits.

An ultra-performance liquid chromatography with tandem mass spectrometric detection (UPLC-MS/MS) method was developed for the detection of flunixin residues in rabbit tissues. The samples were extracted with acidic acetonitrile, defatted with n-hexane, and then purified by HLB solid-phase extraction cartridge. Analysis was carried out on UPLC-ESI-MS/MS working with multiple reaction monitoring (MRM) mode. The limits of detection (LODs) of the method were 0.3-0.8µgkg(-1) and limits of quantification (LOQs) were 1.0-3.0µgkg(-1) in rabbit tissues, respectively. In all fortified samples at a concentration range of 1.0-300.0µgkg(-1), mean recoveries were 61.7-115.7% with relative standard deviations (RSDs) below 16%. Residue depletion of flunixin in rabbit was conducted after oral administration at a dose of 5mgkg(-1) of body weight. The average concentrations for flunixin measured 2h post-administration in kidney and intestine were significantly higher than in liver, heart and muscle. The concentrations for flunixin in all rabbit tissues were below the LOD or not detected in all tissues after 96h administration of drug. A minimum withdrawal time of 21h was indicated for residue levels in heart, liver, kidney, intestine and muscle below the maximum residue limits (MRLs).

Development of a simple method for simultaneous determination of nine subclasses of non-steroidal anti-inflammatory drugs in milk and dairy products by ultra-performance liquid chromatography with tandem mass spectrometry.

A multi-residue analysis method for simultaneous determination of nine subclasses of non-steroidal anti-inflammatory drugs (NSAIDs) in milk and dairy products by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) has been established. The sample was initially extracted and deproteinized with ascorbic acid buffer (0.01M, pH 3) and acetonitrile-ethyl acetate mixture, followed by centrifugation and evaporation, then reconstituted with acetonitrile-0.1% formic acid (1+1, v/v). After removal of lipid material by n-hexane, the sample was analyzed by UPLC-MS/MS with electro-spray ionization (ESI) interface in Multiple Reaction Monitoring (MRM) mode. The range of limits of detection (LODs) and limits of quantification (LOQs) were 0.03-0.30µg/kg and 0.10-1.00µg/kg, respectively. The recoveries in milk, milk powder, yogurt, processed cheese and milk beverage ranged from 61.7% to 117%, and the relative standard deviations (RSDs) were less than 17.9% at three spiked levels (1, 10 and 100 times of the LOQ). Matrix effects were also investigated and it was determined the signals of the analytes were suppressed from 9.4% to 76.6% in processed cheese. The proposed method was also applied to incurred sample analysis. The results proved that this method was suitable for the simultaneous determination of nine subclasses of NSAIDs residues in milk and dairy products.

Silencing of twist expression by RNA interference suppresses epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer.

PURPOSE: This study aimed to explore the role of the Twist gene in the epithelial-mesenchymal transition of ovarian cancer. METHODS: An RNA interference plasmid expressing a small interfering RNA (siRNA)-targeting Twist (Twist siRNA vector) was designed, constructed, and transfected into the human ovarian cancer cell line A2780. Transfection efficiency was assessed under a fluorescence microscope. Changes in the expression of Twist mRNA in A2780 after transfection with the pGenesil Twist shRNA plasmid were analyzed through RT-PCR. MTT assays and adhesion experiments were applied to determine changes in proliferation and adhesion ability of A2870 after transfection with the Twist shRNA plasmid. Changes in the expression of the E-cadherin and N-cadherin proteins in A2780 after transfection with the Twist shRNA plasmid were analyzed using Western blotting. RESULT: The restructuring plasmid pGenesil-Twist shRNA was constructed successfully. After 48 h of culture, 80% of the cells expressed high-intensity GFP fluorescence and stability. The expression of Twist decreased significantly after the transfection of the Twist shRNA plasmid (P<0.05). Proliferation of the transfected Twist shRNA cells showed no difference with that of the A2780-nontransfection or A2780-si-control groups (P>0.05) but the adhesion ability of A2780 decreased dramatically (P<0.05). Expression of the E-cadherin protein increased, whereas that of the N-cadherin protein decreased compared with that in the A2780-nontransfection or A2780- si-control groups (P<0.05). CONCLUSION: Twist is essential for epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer.

Simultaneous determination of thirty non-steroidal anti-inflammatory drug residues in swine muscle by ultra-high-performance liquid chromatography with tandem mass spectrometry.

An ultra-high-performance liquid chromatography with tandem mass spectrometric detection (UHPLC-MS/MS) method was established for the simultaneous determination of residues of thirty non-steroidal anti-inflammatory drugs (NSAIDs) in swine muscle. The samples were extracted with acetonitrile and phosphoric acid. The extracts were defatted with n-hexane, and then purified by HLB solid-phase extraction cartridge. Analysis was carried out on UHPLC-ESI-MS/MS working with multiple reaction monitoring mode with polarity switching. Limits of detection were between 0.4 µg/kg and 2.0 µg/kg, and limits of quantification were between 1.0 µg/kg and 5.0 µg/kg. The recoveries of NSAIDs were between 61.7% and 125.7% at spiked levels of 1.0-500 µg/kg. The repeatability was less than 8% and the within-laboratory reproducibility was not more than 12.3%. The method was reliable, convenient and sensitive.

Gait instability in patients with small acoustic neuroma.

BACKGROUND: Small acoustic neuromas seldom result in typical vestibular symptoms, despite the tumor arising from the vestibular nerve. In this study, we have shown that abnormal gait in eleven patients with small acoustic neuroma could be detected in gait analysis by the use of tactile sensor. Patients displayed no oculomotor abnormality and had tumors less than 10 mm from the porus acoustics. METHODS: Gait related parameters including the coefficients of variations (CV) of stance, swing, double support, area ratio of trajectories of center of force (TCOF), in addition to the foot pressure difference between both feet, were used for assessment of gait. RESULTS: The CV of swing and the area ratio of TCOF were greater in patients than those in the control group (P < 0.05). The values of these two parameters became greater under an eyes closed condition compared to eyes open (P < 0.05) in the patient group. CONCLUSION: These results indicate that gait analysis may be helpful to assess vestibulospinal function of patients with small acoustic neuroma, the slight vestibular deficits of which can not be detected by visual observation.