Effects of Clarithromycin and Ketoconazole on FK506 Metabolism in Different CYP3A4 Genotype Recombinant Metabolic Enzyme Systems.

OBJECTIVE: This study aimed to investigate the effects of clarithromycin and ketoconazole on the pharmacokinetic properties of tacrolimus in different CYP3A4 genotype recombinant metabolic enzyme systems, so as to understand the drug interactions and their mechanisms further. METHOD: The experiment was divided into three groups: a blank control group, CYP3A4*1 group and CYP3A4*18 recombinant enzyme group. Each group was added with tacrolimus (FK506) of a series of concentrations. Then 1 umol/L clarithromycin or ketoconazole was added to the recombinant enzyme group and incubated in the NADPH system for 30 minutes to examine the effects of clarithromycin and ketoconazole on the metabolizing enzymes' activity of different genotypes. The remaining concentration of FK506 in the reaction system was determined using UPLC-MS/MS, and the enzyme kinetic parameters were calculated using the software. RESULTS: The metabolism of CYP3A4*18 to FK506 was greater than that of CyP3А4*1B. Compared with the CYP3A4*1 group, the metabolic rate and clearance of FK506 in the CYP3A4*18 group significantly increased, with Km decreasing. Clarithromycin and ketoconazole inhibit the metabolism of FK506 by affecting the enzyme activity of CYP3A4*1B and CYP3A4*18B. After adding clarithromycin or ketoconazole, the metabolic rate of FK506 significantly decreased in CYP3A4*1 and CYP3A4*18, with Km increasing, Vmax and Clint decreasing. CONCLUSION: Compared with CYP3A4*1, CYP3A4*18 has a greater metabolism of FK506, clarithromycin and ketoconazole can inhibit both the enzymatic activities of CYP3A4*1 and CYP3A4*18, consequently affecting the metabolism of FK506 and the inhibitory on CYP3A4*1 is stronger.

Case report: A rare multidrug-resistant Escherichia coli causes fatal neonatal meningoencephalitis.

Neonatal meningitis is rare but devastating disease. Multidrug-resistant (MDR, multi-drug resistant) bacteria are a major global health risk. We report an Escherichia coli meningitis isolate with multiple resistance patterns and unusual serotype (O75) that caused sudden neonatal death. The isolate was resistant to antibiotics other than cefoperazone/sulbactam and imipenem, challenging the combination of antibiotics commonly used in the empirical treatment of neonatal sepsis. Despite aggressive symptomatic and supportive treatment of the infant based on laboratory tests and clinical practice, the infant eventually died. This is the first case of meningoencephalitis due to serotype O75 reported in China. The presence of highly pathogenic multidrug-resistant microorganisms isolated in neonates underscores the need to implement rapid resistance diagnostic methods and should prompt consideration of alternatives to empiric treatment of neonatal bacterial meningitis.

Anomalous ionic transport in tunable angstrom-size water films on silica.

Liquid and ionic transport through nanometric structures is central to many phenomena, ranging from cellular exchanges to water resource management or green energy conversion. While pushing down toward molecular scales progressively unveils novel transport behaviors, reaching ultimate confinement in controlled systems remains challenging and has often involved 2D Van der Waals materials. Here, we propose an alternative route which circumvents demanding nanofabrication steps, partially releases material constraints, and offers continuously tunable molecular confinement. This soft-matter-inspired approach is based on the spontaneous formation of a molecularly thin liquid film onto fully wettable substrates in contact with the vapor phase of the liquid. Using silicon dioxide substrates, water films ranging from angstrom to nanometric thicknesses are formed in this manner, and ionic transport within the film can then be measured. Performing conductance measurements as a function of confinement in these ultimate regimes reveals a one-molecule thick layer of fully hindered transport nearby the silica, above which continuum, bulk-like approaches account for experimental results. Overall, this work paves the way for future investigations of molecular scale nanofluidics and provides insights into ionic transport nearby high surface energy materials such as natural rocks and clays, building concretes, or nanoscale silica membranes used for separation and filtering.

Dental Materials Applied to 3D and 4D Printing Technologies: A Review.

As computer-aided design and computer-aided manufacturing (CAD/CAM) technologies have matured, three-dimensional (3D) printing materials suitable for dentistry have attracted considerable research interest, owing to their high efficiency and low cost for clinical treatment. Three-dimensional printing technology, also known as additive manufacturing, has developed rapidly over the last forty years, with gradual application in various fields from industry to dental sciences. Four-dimensional (4D) printing, defined as the fabrication of complex spontaneous structures that change over time in response to external stimuli in expected ways, includes the increasingly popular bioprinting. Existing 3D printing materials have varied characteristics and scopes of application; therefore, categorization is required. This review aims to classify, summarize, and discuss dental materials for 3D printing and 4D printing from a clinical perspective. Based on these, this review describes four major materials, i.e., polymers, metals, ceramics, and biomaterials. The manufacturing process of 3D printing and 4D printing materials, their characteristics, applicable printing technologies, and clinical application scope are described in detail. Furthermore, the development of composite materials for 3D printing is the main focus of future research, as combining multiple materials can improve the materials' properties. Updates in material sciences play important roles in dentistry; hence, the emergence of newer materials are expected to promote further innovations in dentistry.

Effects of clarithromycin on the pharmacokinetics of tacrolimus and expression of CYP3A4 and P-glycoprotein in rats.

The objective of this study was to investigate the effect of clarithromycin on the pharmacokinetics of tacrolimus in rats and better understand its mechanism. In the control group (n = 6), rats received a single oral dose of 1 mg tacrolimus on day 6. In the experimental group (n = 6), rats received 0.25 g of clarithromycin daily for five consecutive days and then a single oral dose of 1 mg tacrolimus on day 6. Orbital venous blood (250 µL) was collected at 0, 0.25, 0.50, 0.75, 1, 2, 4, 8, 12, and 24 h before and after tacrolimus administration. Blood drug concentrations were detected via mass spectrometry. Small intestine and liver tissue samples were collected after rats were euthanized via dislocation, and CYP3A4 and P-glycoprotein (P-gp) protein expression was determined using western blotting. Clarithromycin increased the blood tacrolimus concentration and affected its pharmacokinetic properties in rats. Compared with those in the control group, the AUC0-24 , AUC0-∞ , AUMC(0-t) , and AUMC(0-∞) of tacrolimus in the experimental group were significantly increased, whereas the CLz/F was significantly lower (P < 0.01). Simultaneously, clarithromycin significantly inhibited CYP3A4 and P-gp expression in the liver and intestine. Protein expression of CYP3A4 and P-gp in the liver and the intestinal tract was significantly downregulated in the intervention group compared with that in the control group. Clarithromycin significantly inhibited the protein expression of CYP3A4 and P-gp in the liver and intestine, thereby increasing the mean blood concentration and significantly increasing the AUC of tacrolimus.

MSC-EXO and tempol ameliorate bronchopulmonary dysplasia in newborn rats by activating HIF-1α.

BACKGROUND: Bronchopulmonary dysplasia (BPD) is a major complication of premature infants and an important cause of morbidity and mortality. This study investigates the effect of the combination of mesenchymal stem cells-derived exosomes (MSC-EXO) and tempol on BPD and analyzes its mechanism. METHODS: MSC-EXO was extracted by centrifugation and identified by transmission electron microscopy (TEM), nanoparticle tracking analysis, and western blot analysis (WB). Tidal volume (TV), minute ventilation (MV), peak inspiratory flow (PIF), and dynamic pulmonary compliance (Cdyn) of rats were measured by BuxCo pulmonary function experimental platform. Hematoxylin-eosin staining was performed to observe the lung morphology and radical alveolar count (RAC) and mean linear intercept (MLI) were assessed. Immunofluorescence (IF) was conducted to detect the expression of CD31 and α-SMA in pulmonary blood vessels. The kits were used to calculate malondialdehyde (MDA), superoxide dismutase (SOD), and total antioxidant capacity (TAOC) concentration in lung tissue. Enzyme linked immunosorbent assay was applied to detect the levels of IL-1ß, IL-17, IL-6, and IFN-γ in bronchoalveolar lavage fluid. In addition, the expressions of HIF-1α, vascular endothelial growth factor (VEGF), p-PI3K, and p-AKT were analyzed by WB and IF. RESULTS: We successfully extracted and identified MSC-EXO. In BPD rats, TV, MV, PIF, and Cdyn decreased, alveoli were simplified, and the number of interalveoli small vessels, blood vessel density decreased. Moreover, RAC, CD31, TAOC, and SOD decreased, and MLI, α-SMA, MDA, IL-1ß, IL-17, IL-6, and IFN-γ increased, which was reversed by the combination of MSC-EXO and tempol treatment after combined treatment. In addition, the expression levels of HIF-1α, VEGF, p-PI3K, and p-AKT were increased after combined treatment. CONCLUSIONS: Combined treatment could improve lung tissue injury, promote pulmonary vascular remodeling, restore lung function, and inhibit oxidative stress in BPD rats. These effects were achieved through activation of HIF-1α.

Lithium Cholesterol Sulfate: A Novel and Potential Drug for Treating Alzheimer's Disease and Autism Spectrum Disorder.

BACKGROUND AND OBJECTIVE: Recent studies have shown that lithium treatment can reduce symptoms of Alzheimer's disease (AD) and Autism Spectrum Disorder (ASD). However, the present lithium salts clinically available have serious short-term and long-term side effects which requires frequent monitoring of blood chemistry and plasma lithium levels so as to avoid toxicity. Consequently, there is a demand for a safer and more effective lithium formulation to treat these diseases. METHODS: Hence, we firstly synthesized lithium cholesterol sulfate (LiCS) and compared its pharmacological effects with that of lithium chloride (LiCl) and sodium cholesterol sulfate (NaCS) on markers of neurodegenerative disease in cell cultures. RESULTS: LiCS was more potent than LiCl in increasing inhibitory GSK3ß (Ser9) phosphorylation (pGSK3ß) in both CHO and SH-SY5Y cells. These agents dose-dependently increased pGSK3ß, starting at 10 µM for LiCS and 60µM for LiCl and maximally by approximately 100% at 60 µM for LiCS and 1.25 mM for LiCl, without altering total GSK3ß levels. In HEK293/tau cells, LiCS reduced tau (Thr231) phosphorylation (ptau) starting at 10 µM and maximally by 63% at 40 µM without altering total tau levels, but ptau levels were not altered by LiCl at any dose between 60 µM and 1.25 mM. In BV2 cells, LiCS and LiCl decreased LPS-induced TNFα levels, starting at 20 µM for LiCS and 5 mM for LiCl, and maximally by approximately 30% at 80 µM for LiCS and 20 mM for LiCl. NaCS at any dose between 5 and 90 µM did not alter pGSK3ß, ptau or LPS-induced TNFα. CONCLUSION: LiCS may become a new drug with good pharmacological potential for the treatment of neurodegenerative disorders such as AD and ASD by allowing lithium to more readily access intracellular pathological processes.

An overview of three-dimensional imaging devices in dentistry.

OBJECTIVE: To review four types of three-dimensional imaging devices: intraoral scanners, extraoral scanners, cone-beam computed tomography (CBCT), and facial scanners, in terms of their development, technologies, advantages, disadvantages, accuracy, influencing factors, and applications in dentistry. METHODS: PubMed (National Library of Medicine) and Google Scholar databases were searched. Additionally, the scanner manufacturers' websites were accessed to obtain relevant data. Four authors independently selected the articles, books, and websites. To exclude duplicates and scrutinize the data, they were uploaded to Mendeley Data. In total, 135 articles, two books, and 17 websites were included. RESULTS: Research and clinical practice have shown that oral and facial scanners and CBCT can be used widely in various areas of dentistry with high accuracy. CONCLUSION: Although further advancement of these devices is desirable, there is no doubt that digital technology represents the future of dentistry. Furthermore, the combined use of different devices may bring dentistry into a new era. These four devices will play a significant role in clinical utility with high accuracy. The combined use of these devices should be explored further. CLINICAL SIGNIFICANCE: The four devices will play a significant role in clinical use with high accuracy. The combined use of these devices should be explored further.

OATP1B1 Plays an Important Role in the Transport and Treatment Efficacy of Sorafenib in Hepatocellular Carcinoma.

BACKGROUND: Sorafenib is an anticancer drug used in the treatment of unresectable hepatocellular carcinoma and advanced renal cell carcinoma. It is a substrate for the human OATP1B1. This study is aimed at assessing the role of OATP1B1 in transportation and uptake of sorafenib in hepatocellular carcinoma and how OATP1B1 affects the pharmacodynamics of sorafenib in vitro and in vivo. METHODS: Sorafenib transport was measured in HepG2, HepG2-OATP1B1∗1a, HepG2-OATP1B1∗1b, HepG2-OATP1B1∗15, LO2, LO2-OATP1B1∗1a, LO2-OATP1B1∗1b, and LO2-OATP1B1∗15 cells, as well as in HepG2 cells transfected with miR-148a mimics. The viability and apoptosis rate of cells treated with sorafenib were evaluated. A liver cancer rat model was established to explore the pharmacokinetics and pharmacodynamics of sorafenib after overexpression of Oatp2. RESULTS: Changes in expression and genetic mutations of OATP1B1 significantly affected the uptake of sorafenib in HepG2 and LO2 transgenic cells, and the uptake of sorafenib was higher in HepG2 than LO2. Genetic mutations of OATP1B1 significantly affected the cell viability and apoptosis rate of HepG2 cells after sorafenib treatment. Compared to control group, the uptake of sorafenib in miR-148a mimic-transfected HepG2 cells was decreased, and the cell viability was increased. PCN significantly increased the expression of Oatp2 and affected the pharmacokinetics of sorafenib. Vascular endothelial growth factor levels and microvascular density in tumor-adjacent tissues decreased significantly, suggesting that increased Oatp2 expression improves the treatment effect of sorafenib in a rat model of liver cancer. CONCLUSIONS: OATP1B1 plays an important role in the pharmacokinetics and pharmacodynamics of sorafenib in hepatocellular carcinoma.

Uptake of Aß by OATPs might be a new pathophysiological mechanism of Alzheimer disease.

BACKGROUND: The accumulation of neurotoxic amyloid-beta (Aß) in the brain is a characteristic of Alzheimer's disease (AD), at the same time, it is possible alterations of liver function could affect brain Aß levels through changes in blood Aß concentration. Over the last decade, a number of reports have shown that P-glycoprotein (encoded by ABC1B1) actively mediates the efflux transport of Aß peptides. However, the mechanism by which Aß peptides enter the cells is not clear. In the preliminary study, we found that the protein expression of organic anion transporting Polypeptide 1a4 (OATP1B1) in the liver tissue of mice with AD was significantly higher than that in the normal mice. In contrast, the protein expression of Oatp1a4 in the brain significantly decreased in mice with AD. OATP1B1, an important drug transporter might be related to the pathophysiology of AD. RESULTS: In this study, we established an OATP1B1-GFP-HEK293T cell model to confirm the OATP1B1 mediated transport of Aß1-42. Compared to the control group of GFP-HEK293Tcells, the uptake of Aß1-42 protein in the OATP1B1-GFP-HEK293T group increased significantly with the increase in concentration of Aß1-42, and also increased significantly with an increase in the duration of incubation. Similar results were observed in the flow cytometry experiment, and the uptake of Aß1-42in HEK293T-OATP1B1 cells was almost twice that in the control group. These results indicate that OATPs may act as an important "carrier" for the transport of Aß1-42 from the blood to the tissues, including liver and brain. CONCLUSIONS: This is a novel and interesting finding and OATP1B1 can be investigated as a new treatment target for AD.

Altered gut microbiota correlated with systemic inflammation in children with Kawasaki disease.

Kawasaki disease (KD) is a multi-systemic vasculitis of unknown etiology that occurs mainly in children, and the disturbance of gut microbiota is generally believed to cause a hyperimmune reaction triggering KD. The aim of the study was to investigate the alterations in the fecal microbiota and assess its relationship with systemic inflammation. Totally 30 KD children were enrolled and followed up for 6 months, with another group of 30 age- and sex-matched healthy children as controls. Phylotype profiles of fecal microbial communities were analyzed using 16S rRNA gene sequencing. Serum inflammatory markers were detected by flow cytometer. We showed that KD children exhibited a significant reduction in fecal microbial diversity in the acute phase compared with the healthy controls. Enterococcus, Acinetobacter, Helicobacter, Lactococcus, Staphylococcus and Butyricimonas in acute KD children were significantly higher than the healthy children. Levels of systemic inflammation biomarkers, including IL-2, IL-4, IL-6, IL-10, TNF-α, and INF-γ, were significantly elevated in the acute KD children. Altered microbiota genera Enterococcus and Helicobacter abundances were shown to be correlated positively with IL-6, which were never previously reported in KD. This study suggested that gut microbiota alteration is closely associated with systemic inflammation, which provides a new perspective on the etiology and pathogenesis of KD.

Tumour necrosis factor-α and myoglobin associated with the recovery time of coronary artery lesions in Kawasaki disease patients.

AIM: To assess the relationship between clinical parameters and medium term recovery time of coronary artery lesions (CALs). METHODS: In total, 344 Kawasaki disease patients were screened and 311 Kawasaki disease patients were included and followed-up for the next 2 years. Clinical records, clinical parameters and inflammatory biomarkers were collected for all subjects. RESULTS: Tumour necrosis factor (TNF)-α and myoglobin (MYO) levels in patients without recovery from CALs were significantly higher than those without CALs and with recovery from CALs. Kaplan-Meier survival analysis showed that in the high-TNF-α group, the estimated median time to recovery (5.0 months, 95% confidence interval (CI) 1.436-8.564) is significantly longer than the low-TNF-α group (2.00 months, 95% CI: 0.633-3.367, P = 0.044). Also, the estimated median time (5.0 months, 95% CI: 1.836-8.164) in the high-MYO group is significantly longer than the low-MYO group (2.00 months, 95% CI: 0.405-3.595, P = 0.002). Cox regression analysis showed independent factors for recovery of CALs included age, left coronary artery to aortic annulus ratio, TNF-α and MYO levels. CONCLUSIONS: These findings suggest that clinical parameters such as age, left coronary artery to aortic annulus ratio, TNF-α and MYO levels associate with medium term recovery time of CALs and could help in the design of a clinical strategy for the surveillance and prevention of late cardiovascular events.

Expression of Oatp2 in the Brain and Liver of Alzheimer Disease Mouse Model.

The purpose of this study was to explore the expression of Oatp2 transporter in the liver and brain of Alzheimer's disease (AD) mice. The results showed that the protein expression of Oatp2 in the brain significantly decreased in Alzheimer disease mice. On the contrary, protein expression of Oatp2 in liver tissue of AD mice was significantly higher compared with that in normal mice. For mRNA expression of Oatp2, the results showed that, compared with normal mice, it was significantly lower in the brain but significantly higher in liver tissue. Based on the current research, we cannot confirm the relationship between Oatp2 expression and Alzheimer disease. However, it may be a very novel and interesting discovery and may become a new target for the pathogenesis, drug development, and treatment of AD.

A case report on mother-to-child transmission of Brucella in human, China.

BACKGROUND: Human brucellosis is endemic in China and commonly occurs through contact with infected animals from working with livestock or consumption of unpasteurized dairy products. Although rare, human-to-human, and possible sexual transmission, of Brucella has been reported. In this report, we describe a case of likely mother-to-child transmission of Brucella in Hunan Province, China. CASE PRESENTATION: Between June and October 2016, a 28-year old man sought care for testicular swelling and pain at several health facilities. His 26-year old wife developed intermittent fever along with right thigh and hip pain between November 2016 and February 2017 respectively. On April 5, 2017, the female patient delivered a male neonate at 34 weeks of gestation through natural labor. The child's venal blood sample was cultured on April 5, 2017. Brucella was isolated and identified on April 12, 2017. On the same date, serum antibodies of the father and mother were above 1:100 (based on the serum agglutination test [SAT]). The strains isolated from the mother and neonate were identified as Brucella melitensis biotype 1. CONCLUSIONS: This report highlights a family cluster of brucellosis. Culture results strongly support mother-to-child transmission, and a high probability of sexual transmission from husband to wife.

Effect of intermittent hypoxia or hyperoxia on lung development in preterm rat neonates during constant oxygen therapy.

Impaired lung development is a major negative factor in the survival of preterm neonates. The present study was aimed to investigate the impact of constant oxygen, intermittent hyperoxia, and hypoxia on the lung development in preterm rat neonates. Neonatal rats were exposed to 40% O2 with or without brief hyperoxia episodes (95% O2 ) or brief hypoxia episodes (10% O2 ) from day 0 to day 14, or to room air. The body weight, radical alveolar count (RAC), and total antioxidant capacity (TAOC) were significantly lower whereas the lung coefficient and malondialdehyde (MDA) were significantly higher in the hyperoxia and hypoxia groups than the air control and constant oxygen group at day 7, day 14, and day 21 after birth. The lung function indexes were reduced by intermittent hyperoxia and hypoxia. In contrast, the constant oxygen therapy increased the lung function. HIF-1α and VEGF expression were significantly increased by hypoxia and decreased by hyperoxia. The constant oxygen therapy only decreased the HIF-1α expression at day 14 and 21. In summary, the constant oxygen treatment promoted lung function without affecting the antioxidative capacity in preterm rat neonates. While intermittent hyperoxia and hypoxia inhibited lung development, decreased antioxidative capacity, and dysregulated HIF-1α/VEGF signaling in preterm rat neonates.

Congenital brucellosis in a Chinese preterm neonate: A case report.

Although extremely rare, congenital brucellosis can occur via perinatal transmission. We report a case of an infant born prematurely at 34-36 weeks' gestation who had pyrexia, shortness of breath, hepatosplenomegaly and thrombocytopenia. Blood cultures were positive for Gram-negative coccobacilli and Brucella infection was suspected. While, serological tests were negative for Brucella antibodies, B. melitensis infection was confirmed by polymerase chain reaction (PCR). Serology of the parents' blood confirmed the presence of Brucella. The family did not live in an endemic area but had ridden a camel 12 months before the pregnancy. The bacteria may have been sexually transmitted from father to mother and then to foetus via an intrauterine infection. In endemic areas or where the family has been in close contact with infected animals, brucellosis should be suspected in a severely ill neonate with an unknown infection. Thorough medical histories from the family are essential as early diagnosis and prompt therapy will almost certainly improve neonatal outcome.

Geometrical control of dissipation during the spreading of liquids on soft solids.

Gel layers bound to a rigid substrate are used in cell culture to control differentiation and migration and to lower the friction and tailor the wetting of solids. Their thickness, often considered a negligible parameter, affects cell mechanosensing or the shape of sessile droplets. Here, we show that the adjustment of coating thickness provides control over energy dissipation during the spreading of flowing matter on a gel layer. We combine experiments and theory to provide an analytical description of both the statics and the dynamics of the contact line between the gel, the liquid, and the surrounding atmosphere. We extract from this analysis a hitherto-unknown scaling law that predicts the dynamic contact angle between the three phases as a function of the properties of the coating and the velocity of the contact line. Finally, we show that droplets moving on vertical substrates coated with gel layers having linear thickness gradients drift toward regions of higher energy dissipation. Thus, thickness control opens the opportunity to design a priori the path followed by large droplets moving on gel-coated substrates. Our study shows that thickness is another parameter, besides surface energy and substrate mechanics, to tune the dynamics of liquid spreading and wetting on a compliant coating, with potential applications in dew collection and free-surface flow control.