Preliminary Evaluation of in vivo Reflectance Confocal Microscopy Features of Neonatal Lupus Erythematosus.

INTRODUCTION: Neonatal lupus erythematosus (NLE) is a rare autoimmune disease, which needs to be distinguished from eczema, congenital syphilis, and tinea corporis in newborns. Reflectance confocal microscopy (RCM) could be a helpful noninvasive diagnostic tool, which has been used to evaluate several inflammatory skin conditions. The aim of this study was to describe the RCM characteristics of NLE. METHODS: Eleven NLE patients were included in the study, and all patients were evaluated clinically with RCM. We also evaluated RCM images from 11 eczema patients as controls. RESULTS: Some major key diagnostic features of NLE can be observed by RCM: an enlarged honeycomb pattern (9/11, 81.8%), round-to-oval cyst-like structures were present (6/11, 54.5%), the normal ring-like structures were totally or partially obliterated (11/11, 100%) at the level of the dermo-epidermal junction, medium refractivity collagen fibers that were disorganized (10/11, 90.9%), numerous high refractivity round cells (11/11, 100%) in the dermis. CONCLUSION: RCM allows the visualization of major key diagnostic features of NLE and serves as a complementary diagnostic tool for NLE.

An Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS) Method for Qualifying DAPB in Rat Plasma and Application to Pharmacokinetic Studies.

DAPB, a new molecule including danshensu, borneol, and a mother nucleus of ACEI (Angiotensin-converting enzyme inhibitors), is being developed as an antihypertensive candidate compound. A rapid, accurate, and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established and validated for the determination of DAPB in rat plasma. Chromatographic separation was performed on an Agilent SB-C18 column after protein precipitation by acetonitrile with a mobile phase consisting of acetonitrile and deionized water with 0.02% formic acid and 5 mM NH4F (v/v) at a flow rate of 0.2 mL/min. Quantification was performed using electrospray positive ionization mass spectrometry in the multiple reaction monitoring (MRM) mode. The method was linear over the range of 2-1000 ng/mL. The intra- and inter-day precision was within 12%, with accuracies less than 7%. Stability was within the acceptable limits under various storage and processing conditions. No apparent matrix effect was detected. The validated method was applied to the pre-clinical pharmacokinetic study of DAPB after oral administration of 30 mg/kg and intravenous administration of 6 mg/kg in rats.

Linear alopecia in pediatrics: RCM and dermoscopy reveal diagnostic cues.

BACKGROUND: Alopecia areata (AA), trichotillomania (TM), nevus sebaceous (NS), and linear scleroderma en coup de sabre (LSCS) can all present with a patch of linear alopecia, making diagnosis challenging. The purpose of this study was to combine reflectance confocal microscopy (RCM) and dermoscopy in the diagnosis of these lesions in children. METHODS: A total of 36 patients with linear alopecia were enrolled, of whom 14 had AA, seven had TM, nine had NS, and six had LSCS. We evaluated the characteristics and distinguishing features of the four conditions using RCM and dermoscopy. RESULTS: The key to differential diagnosis was the dermal Hair follicle density in the dermis was decreased in AA, and the size and density of the follicular openings were normal in TM. In NS, the major features were petal-like and frogspawn-like structures. In LSCS, dermal papillary rings, sebaceous glands, and follicles were partially or completely missing, and abundant fibrous material was distributed in the dermis. Dermoscopy revealed alopecia, and all four conditions resulted in decreased hair density. AA patients exhibited yellow dots, black dots, and exclamation mark hairs. TM patients presented with irregularly broken hairs and blood spots. Both NS and LSCS patients exhibited an absence of follicular openings; NS patients demonstrated whitish and yellowish round structures, while an atrophic area with white patches, linear vessels, and no yellow or black dots was observed in LSCS patients CONCLUSION: RCM combined with dermoscopy can provide additional information on disease states and differentiate between AA, TM, NS, and LSCS.

In vivo evaluation of vulvar lichen sclerosus with reflectance confocal microscopy and therapeutic monitoring in children.

BACKGROUND: Vulvar lichen sclerosus (VLS) in girls presents with itching, dysuria, and constipation and may result in the loss of vulvar architecture. In patients with an ambiguous clinical presentation, reflectance confocal microscopy (RCM) could be a helpful noninvasive diagnostic tool. The aim of this study was to describe the RCM characteristics of VLS and explore the clinical application value of RCM in therapeutic monitoring. METHODS: Sixteen patients with VLS were included in the study. All patients were periodically evaluated clinically with RCM, and different treatment regimens were given based on the patient's clinical appearances and RCM features. RESULTS: Some major key diagnostic features of VLS can be observed by RCM, including round to oval cyst-like structures with medium-to-low-refractive keratinoid substances (75%), thinning of the epidermal thickness (100%), destruction of the ring-like structures around dermal papillae (100%), disorderly distributed coarse medium-refractive fibrous material (100%),polygonal, plump, high-refractive cellular structures and linear low-refractive canalicular structures (100%). All of these characteristics had a high correspondence with histopathological features. The clinical manifestations improved after individualized treatment regimens based on the clinical appearances and RCM features. CONCLUSION: RCM allows the visualization of major key diagnostic features of VLS and represents a valid option for objective therapeutic monitoring.

Chemical Synthesis, Safety and Efficacy of Antihypertensive Candidate Drug 221s (2,9).

Hypertension is the main risk factor of cardiovascular and cerebrovascular diseases. In this paper, a novel compound known as 221s (2,9), which includes tanshinol, borneol and a mother nucleus of ACEI, was synthesized by condensation esterification, deprotection, amidation, deprotection, and amidation, with borneol as the initial raw material, using the strategy of combinatorial molecular chemistry. The structure of the compound was confirmed by 1H NMR, 13C NMR, and high-resolution mass spectrometry, with a purity of more than 99.5%. The compound 221s (2,9) can significantly reduce the systolic and diastolic blood pressure of SHR rats by about 50 mmHg and 35 mmHg after 4 weeks of administration. The antihypertensive effect of 221s (2,9) is equivalent to that of captopril. The use of 221s (2,9) can reduce the content of Ren, Ang II and ACE in the serum of SHR rats, inhibit the RAAS and enhance the vascular endothelial function by upregulating the level of NO. Pathological studies in this area have shown that high dosage of 221s (2,9) can notably protect myocardial fibrosis in rats and reduce the degeneration and necrosis of myocardial fibers, inflammatory cell infiltration, and proliferation of fibrous tissue in the heart of rat. Therefore, the existing work provided a foundation for preclinical research and follow-up clinical research of 221s (2,9) as a new drug.

In vivo evaluation of facial papule dermatoses with reflectance confocal microscopy in children.

BACKGROUND: Molluscum contagiosum (MC), milia, keratosis pilaris (KP), verruca plana (VP), seborrheic keratosis (SK), and juvenile xanthogranuloma (JXG) are common papule dermatoses on the face of children that have a similar appearance. In vivo evaluation of facial papule dermatoses with reflectance confocal microscopy (RCM) is helpful in the diagnosis of these ambiguous lesions in children. The purpose of this study was to clarify the RCM characteristics of MC, milia, KP, VP, SK, and JXG and explore the clinical application value of RCM for these common facial papule dermatoses. METHODS: We recruited 113 patients referred for unequivocal facial papule dermatosis, including 21 patients with MC, 17 patients with milia, 19 patients with KP, 36 patients with VP, 8 patients with SK, and 12 patients with JXG. We evaluated the characteristics and distinguishing features of the six kinds of facial papule dermatoses using RCM. RESULTS: The main RCM features of the six dermatoses included a well-demarcated border of the lesion area. MC, milia and KP all manifested cyst-like structures, and their distinguishing features were the location of the cystic structures and the refractive index of the contents. Although VP, SK, and JXG did not have obvious cystoid structures, VP was typically characterized by uniformly distributed petal-like structures with a medium-to-high refractive index in the epidermis. With regard to SK, the characteristic features were an obviously thickened epidermis and cobblestone-like structures. JXG was mainly characterized by multiple large round and ovoid cells with a foamy cytoplasm, and discoid-shaped multinucleated large cells were diffusely distributed in the dermis. CONCLUSION: RCM allows the real-time visualization of major key diagnostic and distinguishing features of common facial papule dermatoses in children, including MC, milia, KP, VP, SK, and JXG.

Voltammetric aptasensor for thrombin by using a gold microelectrode modified with graphene oxide decorated with silver nanoparticles.

A sensitive electrochemical aptasensor was developed for the determination of thrombin. It is based on signal amplification by using graphene oxide (GO) decorated with silver nanoparticles (AgNP) and placed on a microelectrode. The graphene oxide decorated with silver nanoparticles (AgNP-GO) was synthesized by chemical reduction and then modified with a thrombin-binding aptamer 1 (TBA1; a 29-mer) carrying a 3'-terminal thiol group. It was immobilized on the AgNP-GO by self-assembled monolayer technique to form the signaling probe (TBA1-AgNP-GO). A capture thrombin-binding aptamer 2 (TBA-2; a 15-mer) carrying a 3'-terminal thiol group was self-assembled onto the surface of the gold electrode. After binding of thrombin and the signaling probe, a sandwich of type TBA2-thrombin-TBA1-AgNP-GO is formed. The square wave voltammetric signal of the AgNP is then recorded. The electrochemical oxidation signal (measured at 0.21 V vs. Ag/AgCl) increases linearly with the concentration of thrombin in the range from 0.05 nmol·L-1 to 5 nmol·L-1 with a detection limit of 0.03 nmol·L-1 (at S/N = 3). The aptasensor is highly specific and not affected by other proteins. It was successfully applied to the determination of thrombin in spiked human serum samples. Graphical abstract A sensitive electrochemical aptasensor for thrombin was developed based on the signal amplification of silver nanoparticles decorated graphene oxide at an ultramicroelectrode.

Reflectance confocal microscopy features of vulvar lichen sclerosus in Chinese juvenile girls.

Vulvar lichen sclerosus (VLS) is a chronic inflammatory dermatosis of unknown pathogenesis, characterized by porcelain-white atrophic plaques around the vulvar and anal areas in girls. With this communication, we performed the study on 16 female girls with clinically and histologically confirmed VLS, described the main identifying characteristics of the lesions in reflectance confocal microscopy (RCM) and elucidated the corresponding relationship between RCM findings and histology. We recommend RCM, a noninvasive technique, as a complementary diagnostic tool for VLS.

Evaluation of Solubility, Physicochemical Properties, and Cytotoxicity of Naproxen-Based Ionic Liquids.

To solve the problems associated with poorly water-soluble nonsteroidal anti-inflammatory drugs (NSAIDs), naproxen-based ionic liquids (ILs) containing naproxen as an active pharmaceutical ingredient (API) anion were prepared with benzalkonium (tetradecyldimethylbenzyl ammonium), choline, and 1-octyl-3-methylimidazole as the cation. The structures and thermal properties were analyzed. Through the conductivity method, the solubility at 25 and 37 °C and the critical micelle concentration (CMC) at 25 °C were determined in water and ethanol. The octanol-water partition coefficients (K ow) at 25 °C were measured with the shake-flask method. The cytotoxicity was evaluated with the MTT method. The results showed that the conversion of naproxen into the API-ILs increased the API's solubility in water by more than 850 times compared with the original API, and the thermostability was satisfactory with a lower glass transition temperature (t g). Moreover, the variation trends of solubility, hydrophilicity, and K ow were consistent with the different structures of naproxen-based ILs, except for benzalkonium naproxen. The CMC (10-5-10-6 M) in water and ethanol demonstrated that the naproxen-based ILs were surface activite ILs. The IC50 values exhibited the low cytotoxicity of the naproxen-based ILs, which was better than 100 µM. The results provide essential information and a research basis for future topical and transdermal administration and oral administration of naproxen-based ILs.

A functional intact SUMOylation machinery in Aspergillus flavus contributes to fungal and aflatoxin contamination of food.

SUMO adducts occur in Aspergillus flavus, and are implicated in fungal biology, while the underlying mechanism and the SUMOylation apparatus components in this saprophytic food spoilage mould, remain undefined. Herein, genes encoding SUMOylation cascade enzymes in A. flavus, including two heterodimeric SUMO E1 activating enzymes, a unique SUMO E2 conjugating enzyme, and one of SUMO E3 ligases, were identified and functionally analyzed. Global SUMO adducts immunoassay, multiple morphological comparison, aflatoxin attributes test, fungal infection and transcriptomic analyses collectively revealed that: E1 and E2 were essential for intracellular SUMOylation, and contributed to both stress response and fungal virulence-related events, including sporulation, colonization, aflatoxins biosynthesis; the primary E3 in this fungus, AfSizA, might serve as the molecular linkage of SUMOylation pathway to fungal virulence rather than SUMOylation-mediated stress adaptation. These findings demonstrated that SUMOylation machinery in A. flavus was functionally intact and contributed to multiple pathobiological processes, hence offering ideas and targets to control food contamination by this mycotoxigenic fungus.

N-doped carbon dots coupled with molecularly imprinted polymers as a fluorescent sensor for ultrasensitive detection of genistein in soya products.

Rapid detection of genistein in soya products has remained difficult. Current methods necessitate sample handling and use of costly instruments. Here, using a simple one-pot reverse microemulsion method, a sensor based on N-doped carbon dots conjugated molecularly imprinted polymers (N-CDs@MIPs) was synthesized to analyze genistein. N-doped carbon dots were used as fluorescent component, genistein as the template molecule, and molecularly imprinted polymers as the selective sorbent in this fluorescence sensor. The sensor was then examined and optical studies demonstrated that N-CDs@MIPs not only had strong fluorescence emission and outstanding optical stability, but also had good sensitivity (detection limit 35.7 nM) and selectivity to genistein. Furthermore, the N-CDs@MIPs materials were used to analyze genistein in soya products, and the findings (which ranged from 99.77% to 106.11%) show that the N-CDs@MIPs has high potential for quickly detecting the amount of genistein in complicated food samples.

PM2.5 causes vascular hyperreactivity through the upregulation of the thromboxane A2 receptor and activation of MAPK pathways.

Airborne fine particulate matter (PM2.5) is a major cardiovascular disease environmental risk factor. However, the underlying mechanism of action is not fully understood. Thromboxane is widely known as an important vasoconstrictor substance that binds to G-protein-coupled receptors (GPCR) in arteries and is involved in various cardiovascular diseases. This study examined the effect of PM2.5 on thromboxane A2 receptor (TP) in the mesenteric arteries and the underlying intracellular signal mechanisms (by focusing on the mitogen-activated protein kinase (MAPK) pathway). Rat mesenteric artery segments were exposed to PM2.5 in the presence of MAPK pathway inhibitors. The contractile reactivity of mesenteric arteries was analyzed using wire myography. The mRNA and protein expression of TP receptor and MAPK pathway molecules were detected by real-time PCR and Western blot. Mesenteric artery receptor localization was assessed by immunohistochemistry. The results showed that TP receptor-mediated maximum contraction response was achieved after exposing arteries to 1.0 µg/mL PM2.5 for 16 h (Emax: 228 ± 16% of K+). Moreover, inhibitor U0126 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), and SP600125 (JNK inhibitor) depressed the increased TP receptor-mediated contractile responses (reduced rage were 17.9 ~ 59.6%). These inhibitors also decreased the increased mRNA expression and protein of the TP receptor induced by PM2.5 (reduced by more than 50% and 46%, respectively). The immunoreactivity of increased TP receptor expression was primarily localized in the cytoplasm. In addition, phosphorylation quantitative analysis showed that in the presence of MAPK inhibitors, the PM2.5-induced phosphorylation of ERK1/2, p38, and JNK protein increased by more than 30.0 ~ 130.3%. These results suggest that PM2.5 upregulates the TP receptor of rat mesenteric arteries through activation of the ERK1/2, p38, and JNK MAPK pathways.

Protective effects of fisetin against myocardial ischemia/reperfusion injury.

The underlying mechanism of the myocardial protective effect of fisetin was studied in a rat ischemia/reperfusion injury model. Sprague-Dawley rats were randomly assigned to seven groups and pretreated with different solutions by gavage administration. A rat model of cardiac ischemia/reperfusion injury was established. Plasma levels of Von Willebrand factor (vWF) were determined by ELISA, flow cytometry was used to determine the level of cardiomyocyte apoptosis and 2,3,5-triphenyltetrazolium staining was used to determine the size of myocardial infarcts. Hematoxylin and eosin-stained sections of myocardial tissues were examined for pathological changes. Expressions of nuclear factor (NF)-κB and matrix metallopeptidase 9 (MMP-9) were measured by immunohistochemistry. Compared with the model group, rats pretreated with fisetin, quercetin and aspirin showed significant prolongation of clotting time, prothrombin time, thrombin time and activated partial thromboplastin time. Fisetin treatment better maintained the integrity of myocardial fibers and nuclear integrity, reduced the percentage of apoptotic myocardial cells, inhibited expression of NF-κB, decreased the loss of MMP-9 and reduced nuclear translocation of NF-kB. Rats pretreated with fisetin also demonstrated a significant decrease in plasma levels of vWF. In addition, the protective effect of fisetin on myocardial cells was found to be dose dependent.

Specific enrichment of caffeic acid from Taraxacum mon-golicum Hand.-Mazz. by pH and magnetic dual-responsive molecularly imprinted polymers.

Specific recognition of caffeic acid (CA) from Taraxacum mon-golicum Hand.-Mazz. was successfully performed using a new pH responsive magnetic molecularly imprinted polymers (pH-MMIPs) by simple surface molecular imprinting polymerization. The pH-MMIPs were prepared on the surface of the Fe3O4@SiO2@MPS particles using CA as a template, 2-(dimethylamino) ethyl methacrylate (DMA) as the pH responsive functional monomer, 4-vinylpyridine (4-VP) as an assisting functional monomer, ethylene glycol dimethyl acrylate (EGDMA) as cross-linker, 2,2'-azobisisobutyronitrile (AIBN) as initiator and methanol-H2O (1:1, v/v) as the porogen. The resultant polymers were characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), vibrating sample magnetometry (VSM) and x-ray diffraction (XRD). The adsorption experiments revealed that the pH-MMIPs performed high adsorption ability (11.5 mg g-1) by changing solution pH. Successful selective adsorption of CA was achieved with distribution coefficient of 0.12 and 0.21 towards ferulic acid and chlorogenic acid. Furthermore, pH-MMIPs were employed as adsorbents for extraction and enrichment of CA from Taraxacum mon-golicum Hand.-Mazz. extract. The recoveries of CA in the Taraxacum mon-golicum Hand.-Mazz. ranged from 90.47% to 98.97%. The results proved that the polymers have the potential to provide a selective recognition of CA in complex samples by simple pH regulation.

Bis[4-chloro-2-(imino-meth-yl)phenolato]copper(II) methanol disolvate.

The title compound, [Cu(C(7)H(5)ClNO)(2)]·2CH(3)OH, possesses crystallographic twofold symmetry, with the twofold axis passing through the central Cu(II) ion. The metal centre is coordinated by two O atoms and two N atoms from two symmetry-related Schiff base ligands, forming a slightly distorted cis-CuN(2)O(2) square-planar geometry. The complex mol-ecules are linked via the solvent methanol mol-ecules by O-H⋯O and N-H⋯O hydrogen bonds, leading to the formation of chains along the b axis.

Prednison provokes serum and vasoactive substances in a mice model of immune thrombocytopenia.

OBJECTIVES: The main objective of this study was to investigate the variations of ß-endorphin (ß-EP), vasoactive intestinal peptide (VIP), serotonin (5-HT) and norepinephrine (NE) of immune thrombocytopenia (ITP) mice as well as the regulatory mechanism of prednison. MATERIALS AND METHODS: Sixty BALB/c mice were randomly divided into control group, model group and prednison intervention group. ITP mice model was duplicated by injecting with glycoprotein-antiplatelet serum (GP-APS) except in control group. After ITP disease model was successful established, prednison was used in prednison intervention group. The ß-EP, VIP, 5-HT and NE contents of ITP mice were detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with the values in control group, the detection values of VIP and 5-HT in model group declined, while the detection values of ß-EP and NE increased. Compared with prednison intervention group, the detection values of VIP and 5-HT in model group increased, while the detection values of ß-EP and NE showed no significant change. CONCLUSION: In this study, the ß-EP, VIP, 5-HT and NE contents in ITP mice injected with GP-APS were changed by prednison. It shows that prednison as the first-line therapy for ITP with effective hemostasis function is likely to increasing the contents of VIP and 5-HT. These results suggest the therapeutic value of prednison for the treatment of ITP.

[Effects of electroacupuncture on the proprioception of athletes with functional ankle instability].

OBJECTIVE: To observe the efficacy differences between electroacupuncture and physiotherapy on the proprioception of athletes with functional ankle instability (FAD. METHODS: Fifty athletes with FAI were randomly divided into an electroacupuncture group and a physiotherapy group. The electroacupuncture group was treated with electroacupuncture at Jiexi (ST 41), Kunlun (BL 60), Qiuxu (GB 40) and Ashi acupoints, and the physiotherapy group was treated with low frequency electrical stimulation and infrared radiation at medial malleolus and lateral malleolus, thrice each week for consecutive 8 weeks. The Joint Position Sense: Active (JPSA), Joint Position Sense: Passive (JPSP) and Kinaesthesia (KT) were assessed at the ankle by use of Biodex System-III isokinetic dynamometer. RESULTS: The JPSA of 11.090 +/- 3.1 degrees and the JPSP of 9.67 degrees +/- 2.8 degrees before the treatment reduced to 9.14 degrees +/- 4.0 degrees and 6.89 degrees +/- 3.3 degrees, respectively, after the treatment in the electroacupuncture group, with significant differences in JPSA and JPSP (both P < 0.05), compared with those in the physiotherary group, there were significant differences (both P < 0.05) and with no significant difference in KT (P > 0.05). There was no significant differences in the indices of JPSA, JPSP and KT in the physiotherapy group after 8 weeks than those before treatment (all P > 0.05). CONCLUSION: Electroacupuncture can effectively improve the proprioception of athletes with FAI and achieves a superior efficacy as compared with the conventional physiotherapy.