Type II Crigler-Najjar syndrome: a case report and literature review.

Background: Crigler-Najjar syndrome (CNS) is caused by mutations in uridine 5'-diphosphate glucuronyltransferase (UGT1A1) resulting in enzyme deficiency and hyperbilirubinemia. Type II CNS patients could respond to phenobarbital treatment and survive. This study presents a rare case of type II CNS. Case summary: The proband was a 29-year-old male patient admitted with severe jaundice. A hepatic biopsy showed bullous steatosis of the peri-central veins of the hepatic lobule, sediment of bile pigment, and mild periportal inflammation with normal liver plate structure. The type II CNS was diagnosed by routine genomic sequencing which found that the proband with the Gry71Arg/Tyr486Asp compound heterozygous mutations in the UGT1A1 gene. After treatment with phenobarbital (180 mg/day), his bilirubin levels fluctuated between 100 and 200 µmol/L for 6 months and without severe icterus. Conclusion: Type II CNS could be diagnosed by routine gene sequencing and treated by phenobarbital.

Effects of volume-matched acute exercise on metacognition in late middle-aged adults: The roles of exercise intensity and duration.

Increasing studies have observed the benefit of acute exercise on core executive functions (EFs). However, the effects of high-order EFs (i.e., metacognition) are poorly investigated. Additionally, the beneficial effect varies depending on exercise-related variables (e.g., intensity, duration). Exercise volume, represented by exercise intensity and duration, is one approach to examine the relationship between acute exercise and metacognition. Therefore, this study aims to investigate the effects of acute exercise, involving three volume-matched evaluations, on metacognition in late middle-aged adults. It recruited 40 community-dwelling late middle-aged adults (22 females; Mage=61.05±4.17years). Its counterbalanced and repeated-measure experimental design included a control reading session and three exercise sessions: (1) acute moderate-intensity exercise for 30min, (2) higher-intensity exercise for 16min, and (3) lower-intensity exercise for 40min. After each session, all participants were asked to perform the Tower of London (TOL) task to assess their metacognition related to planning and problem-solving. This study showed that regardless of exercise intensity and duration variation, acute exercise led to higher move-related (i.e., total correct and move scores) and time-related (i.e., total executive and problem-solving times) scores but did not affect total initial time and violation-related scores compared to reading treatment. These findings suggest enhanced performance in metacognition after acute exercise regardless of its intensity and duration variation.

Effect of brief mindfulness and relaxation inductions on anxiety, affect and brain activation in athletes.

The mindfulness-based intervention and psychological skills training are often used for maintaining the mental health or reducing undesirable mental states in athletes. However, their differences in acute effects on mental health and underlying neural mechanism are not well understood. Therefore, the purpose of the current study was to examine the differential effects of brief mindfulness induction (MI) and relaxation induction (RI) on state anxiety, affect and brain activation. Thirty-five track and field athletes were recruited for this study. Using a within-subjects crossover design, participants underwent three conditions that incorporated two 30-min experimental conditions (i.e., MI or RI) and a control condition. State anxiety and affect were assessed before and after intervention, and brain activation (i.e., theta, alpha bands) were recorded by electroencephalography (EEG) during each 30-min condition. Repeated measures analysis of variance revealed that MI and RI similarly reduced state anxiety and negative affect from pre-test to post-test compared to the control condition. In terms of positive affect, there were no significant differences among the three conditions across times. Furthermore, participants exhibited higher frontal theta power during the MI and RI than control condition, whereas no differences in alpha power were observed among conditions. The current study provides initial evidence from an electrophysiological perspective that brief MI and RI both improve the negative psychological states in individual sport athletes through similar neural mechanisms. Nevertheless, the moderating effects of training experiences and long-term interventions on mental state and EEG activity in athletes need further investigation in future studies.

Esophageal chemical clearance and mucosa integrity values in refractory gastroesophageal reflux disease patients with different esophageal dynamics.

OBJECTIVES: Esophageal post-reflux swallow-induced peristaltic wave index (PSPWI) and mean nocturnal baseline impedance (MNBI), novel impedance-based markers of reflux burden, are associated with esophageal dynamics. We aim to investigate the characteristics of PSPWI and MNBI in Chinese refractory gastroesophageal reflux disease (RGERD) patients with different esophageal dynamic changes. MATERIALS AND METHODS: 201 RGERD and 76 functional heartburn patients, undergone off-PPI endoscopy, esophageal manometry and impedance-pH monitoring, were included. Comparisons of conventional and novel impedance-pH metrics were made among different esophageal dynamics groups. Receiver operating-characteristic analyses were utilized to evaluate the diagnostic efficacy of PSPWI and MNBI in differentiating abnormal esophageal dynamics. Correlations were used to investigate their associated factors. RESULTS: PSPWI and MNBI of RGERD with esophagogastric junction (EGJ) injury and esophageal dysmotility were lower than EGJ injury alone or normal dynamics (p < 0.05 for both comparisons). PSPWI with esophageal peristalsis abnormality was lower than EGJ injury (p = 0.049), while MNBI showed no statistical difference. PSPWI, MNBI and their combination have auxiliary diagnostic values for esophageal peristalsis [area under the curves (AUCs): 0.683, 0.656, 0.708)] while only their combination for EGJ injury (AUC: 0.610). And they positively correlated with esophageal motility while negatively correlated with ineffective swallows and acid reflux events. CONCLUSIONS: PSPWI and MNBI, indicating impairment of esophageal chemical clearance and mucosa integrity, were lower in RGERD patients with multiple esophageal dynamic injuries than single injuries or normal dynamics. Moreover, they provided useful contributing information for potential dynamic injuries if manometry has already been found normal or marginal.

Effects of Multi-Component Exercise on Sleep Quality in Middle-Aged Adults.

Sleep is a crucial factor in healthy aging. However, most middle-aged adults experience high levels of sleep disorders. While previous findings have suggested exercise training could benefit the quality of sleep, the effects of multi-component exercise on sleep quality are less examined. Accordingly, the current study aimed to assess the effectiveness of a multi-component exercise program on the quality of sleep among middle-aged adults. Twenty-four middle-aged adults were randomly assigned either to a multi-component exercise (MCE) group or a control group. The participants in the MCE group attended a 90-min session per week for 12 weeks. The control group was instructed to maintain their daily routine for 12 weeks. The primary outcome was the sleep quality evaluated by the Pittsburgh Sleep Quality Index (PSQI). The secondary outcome was physical fitness, including muscular strength and endurance, balance, and flexibility. Regarding sleep quality, the global mean score (p = 028), sleep disturbances (p = 011), and sleep efficiency (p = 035) of the PSQI scores were significantly reduced in the MCE group after the 12-week intervention. Regarding physical fitness, the flexibility of the MCE group improved significantly after the intervention (p = 028), yet, no significant change was observed in the control group. Additionally, the muscular strength of the control group declined significantly after the 12-week period (p = 034). Our results revealed the effectiveness of the MCE intervention in improving sleep quality and physical fitness in middle-aged adults. Further studies using larger sample sizes, objective measures of sleep quality, different types of exercise training, as well as different populations, are warranted to extend our current findings.

Neospora caninum infection activated autophagy of caprine endometrial epithelial cells via mTOR signaling.

Neosporosis, caused by infection with the protozoan parasite Neospora caninum, is one of the main causes of abortion in cattle and small ruminants (e.g., goats), negatively influencing animal health and production costs. The uterus is an adhesion organ of placenta that is important for pregnancy and embryonic development. However, the underlying molecular pathogenic mechanisms of N. caninum in the uterus are still unclear. Autophagy regulates innate and adaptive immunity for eliminating pathogens by xenophagy, while pathogens can manipulate autophagy to facilitate their propagation. To study the role of host cell autophagy during N. caninum infection, a N. caninum infection model in caprine endometrial epithelial cells (EECs) was successfully established. In this in vitro model, N. caninum infection increased the expression of LC3-II (a standard marker for autophagosomes) from 6 h post infection (pi) to 48 h pi and the number of autophagosomes in caprine EECs at 48 h pi. Expression of p62 protein (a classical receptor of autophagy) levels were significantly decreased (P < 0.05) in caprine EECs infected with N. caninum tachyzoites at both 24 h pi and 48 h pi. Enhanced autophagic flux was also detected at 48 h pi in caprine EECs infected with N. caninum tachyzoites by transfecting Ad-mCherry-GFP-LC3B recombinant adenovirus. Treatments using a mechanistic target of rapamycin (mTOR)-specific inhibitor (rapamycin) and an autophagy inhibitor (chloroquine) indicated that cell autophagy induced by N. caninum infection promoted the intracellular propagation of parasite tachyzoites. Further studies showed that N. caninum infection induced autophagy through inhibition of mTOR phosphorylation. To the best of our current knowledge, this is the first study to reveal the role of autophagy during N. caninum infection in caprine EECs, and the findings provided significant information for uncovering mechanisms of abortion and pathogenicity caused by N. caninum infection.

Long non-coding RNA (lncRNA) CYTOR promotes hepatocellular carcinoma proliferation by targeting the microRNA-125a-5p/LASP1 axis.

This study investigated the function of long non-coding RNA (lncRNA) cytoskeleton regulator RNA (CYTOR) in hepatocellular carcinoma (HCC). In HCC, the expression of CYTOR and microRNA (miR)-125a-5p were measured by quantitative real-time PCR (qRT-PCR). The expression of actin skeletal protein 1 (LASP1) was evaluated by Western blot analysis. Flow cytometry assays, transwell assays, colony formation assay, and cell counting kit-8 (CCK-8) assay were used to evaluate the roles of miR-125a-5p and CYTOR in HCC cells. The target genes of CYTOR and miR-125a-5p were identified by bioinformatics analysis and Luciferase assay. CYTOR was upregulated in HCC cell lines, and knockdown of CYTOR inhibited HCC cell growth. MiR-125a-5p was downregulated in HCC cells and a target of CYTOR in regulating HCC progression. Furthermore, LASP1 was a downstream target of miR-125a-5p. Finally, CYTOR was found to be involved in HCC progression in vivo. CYTOR promotes HCC development by regulating the miR-125a-5p/LASP1 axis.

Diagnostic Value of Circulating Antigens in the Serum of Piglets with Experimental Acute Toxoplasmosis.

Toxoplasmosis, caused by Toxoplasma gondii, an apicomplexan parasite, infects all warm-blooded animals, including a third of the human population. Laboratory diagnosis of acute toxoplasmosis is based on the detection of anti-T. gondii IgM and IgG and T. gondii nucleic acid; however, these assays have certain limitations. Circulating Ags (CAgs) are reliable diagnostic indicators of acute infection. In this study, we established a model of acute T. gondii infection in Large White pigs. CAg levels peaked between 3 and 5 d after inoculation, and 28 CAgs were identified using an immunoprecipitation-shotgun approach, among which dolichol-phosphate-mannose synthase family protein (TgDPM), C3HC zinc finger-like protein (TgZFLP3), and ribosomal protein RPL7 (TgRPL7) were selected to further investigate their value in the diagnosis of acute toxoplasmosis. Immunofluorescence assays revealed that TgDPM and TgRPL7 were localized in the membrane surface, while TgZFLP3 was localized in the apical end. Western blotting revealed the presence of the three proteins in the serum during acute infection. Indirect ELISA results indicate that TgZFLP3 is likely to be a novel candidate for the diagnosis of acute toxoplasmosis. However, these three proteins may not be useful as candidate vaccines against toxoplasmosis owing to their low protective ability. In addition, deletion of the zflp3 gene partially attenuated virulence in Kunming mice. Collectively, we identified 28 CAgs in the serum of piglets with experimental acute toxoplasmosis and confirmed that TgZFLP3 is a potential biomarker for acute T. gondii infection. The results of this study provide data to improve the detection efficiency of acute toxoplasmosis.

Genome-Wide CRISPR/Cas9 Screen Identifies New Genes Critical for Defense Against Oxidant Stress in Toxoplasma gondii.

Toxoplasma gondii is one of the most widespread apicomplexans and can cause serious infections in humans and animals. Its antioxidant system plays an important role in defending against oxidant stress imposed by the host. Some genes encoding the antioxidant enzymes of T. gondii have been identified; however, critical genes that function in response to reactive oxygen species (ROS) stress are still poorly understood. Here, we performed genome-wide CRISPR/Cas9 loss-of-function screening in the T. gondii RH strain to identify potential genes contributing to the ROS stress response. Under hydrogen peroxide treatment, 30 single guide RNAs targeting high-confidence genes were identified, including some known important antioxidant genes such as catalase and peroxiredoxin PRX3. In addition, several previously uncharacterized genes were identified, among which five hypothetical protein-coding genes, namely, HP1-HP5, were selected for further functional characterization. Targeted deletion of HP1 in T. gondii RH led to significant sensitivity to H2O2, suggesting that HP1 is critical for oxidative stress management. Furthermore, loss of HP1 led to decreased antioxidant capacity, invasion efficiency, and proliferation in vitro. In vivo results also revealed that the survival time of mice infected with the HP1-KO strain was significantly prolonged relative to that of mice infected with the wild-type strain. Altogether, these findings demonstrate that the CRISPR/Cas9 system can be used to identify potential genes critical for oxidative stress management. Furthermore, HP1 may confer protection against oxidative damage and contributes to T. gondii virulence in mice.

MicroRNA-140-5p is Downregulated in Osteosarcoma and Overexpression of MicroRNA-140-5p Inhibits Cancer Cell Proliferation by Downregulating GLUT-1.

BACKGROUND: MicroRNA-140-5p plays pivotal role in different types of human malignancies, while its involvement in osteosarcoma is unknown. OBJECTIVE: Our study aimed to investigate the functionality of microRNA-140-5p in osteosarcoma. METHODS: Plasma levels of microRNA-140-5p and glucose transporter 1 (GLUT-1) in both osteosarcoma and healthy controls were measured by qRT-PCR and ELISA, respectively. Correlation between plasma levels of microRNA-140-5p and GLUT-1 was analyzed by Pearson correlation coefficient. Correlation between plasma levels of microRNA-140-5p and clinical data of patients with osteosarcoma was analyzed by Chi-square test. MicroRNA-140-5p mimic and GLUT-1 expression vector were transfected into cells of human osteosarcoma cell lines, and the effects on microRNA-140-5p expression, GLUT-1 expression and cell proliferation were analyzed by qRT-PCR, Western-blot and CCK-8 assay, respectively. RESULTS: Plasma levels of microRNA-140-5p were significantly lower and plasma levels of GLUT-1 were significantly higher in osteosarcoma patients than that in healthy controls. Levels of plasma microRNA-140-5p and GLUT-1 were reversely correlated in osteosarcoma patients. Plasma levels of microRNA-140-5p were correlated with tumor size but not with other clinical data of patients. MicroRNA-140-5p mimic significantly inhibited cancer cell proliferation, while GLUT-1 overexpression significantly promoted cancer cell proliferation. MicroRNA-140-5p mimic significantly downregulated GLUT-1 expression. CONCLUSION: GLUT-1 overexpression showed no significant effect on microRNA-140-5p expression but attenuated the inhibitory effects of microRNA-140-5p mimic on cell proliferation. We therefore conclude that microRNA-140-5p is downregulated in osteosarcoma and overexpression of microRNA-140-5p may inhibit cancer cell proliferation by downregulating GLUT-1.

Study on Circulating Antigens in Serum of Mice With Experimental Acute Toxoplasmosis.

Toxoplasma gondii is a ubiquitous apicomplexan protozoan parasite that can infect all warm-blooded animals, causing toxoplasmosis. Thus, efficient diagnosis methods for acute T. gondii infection are essential for its management. Circulating antigens (CAgs) are reliable diagnostic indicators of acute infection. In this study, we established a mouse model of acute T. gondii infection and explored new potential diagnostic factors. CAgs levels peaked 60 h after T. gondii inoculation and 31 CAgs were identified by immunoprecipitation-liquid chromatography-tandem mass spectrometry, among which RuvB-like helicase (TgRuvBL1), ribonuclease (TgRNaseH1), and ribosomal protein RPS2 (TgRPS2) were selected for prokaryotic expression. Polyclonal antibodies against these three proteins were prepared. Results from indirect enzyme-linked immunosorbent assay indicated that anti-rTgRuvBL1, anti-rTgRNase H1, and anti-rTgRPS2 mouse sera were recognized by natural excretory-secretory antigens from T. gondii tachyzoites. Moreover, immunofluorescence assays revealed that TgRuvBL1 was localized in the nucleus, while TgRNase H1 and TgRPS2 were in the apical end. Western blotting data confirmed the presence of the three proteins in the sera of the infected mice. Moreover, mice immunized with rTgRuvBL1 (10.0 ± 0.30 days), TgRNaseH1 (9.67 ± 0.14 days), or rTgRPS2 (11.5 ± 0.34 days) had slightly longer lifespan when challenged with a virulent T. gondii RH strain. Altogether, these findings indicate that these three proteins can potentially be diagnostic candidates for acute toxoplasmosis. However, they hold poor protective potential against highly virulent T. gondii infection.

Biomimetic poly(glycerol sebacate)/polycaprolactone blend scaffolds for cartilage tissue engineering.

Poly (glycerol sebacate) (PGS) is a synthetic polymeric material with the characteristics of controllable degradation, high plasticity and excellent biocompatibility. However, the time of PGS degradation is faster than that of cartilage regeneration, which limits its application in cartilage tissue engineering. Polycaprolactone (PCL), a widely used synthetic polymer, has appropriate biodegradability and higher mechanical strength. This study aims to make a scaffold from blends of fast degrading PGS and slowly degrading PCL, and to investigate its potential for cartilage tissue engineering applications. Scanning electron microscopic analysis indicated that the scaffolds provided favourable porous microstructures. In vitro degradation test showed that PGS/ PCL scaffolds acquired longer degradation time and better mechanical strength. PGS/PCL scaffolds seeded with Bone marrow-derived mesenchymal stem cells (BMSCs) and articular chondrocytes (ACCs) were cultured in vitro. Short-term in vitro experiments confirmed that both seeded cells could adhere and proliferate on the scaffold. Chondrogenic culture for cell-scaffold constructs confirmed BMSCs could differentiate into chondrocyte-like cells in PGS/PCL scaffolds. With tunable biodegradation, favorable mechanical properties and cytocompatibility, PGS/PCL scaffolds would potentially be suitable for the regeneration of cartilage tissue. Poly (glycerol sebacate) (PGS) is a synthetic polymeric material with the characteristics of controllable degradation, high plasticity and good biocompatibility. However, the time of PGS degradation is faster than that of cartilage regeneration, which limits its application in cartilage tissue engineering. Polycaprolactone(PCL), a widely used synthetic polymer, has appropriate biodegradability. This study aims to make a scaffold from blends of fast degrading PGS and slowly degrading PCL, and to investigate its potential for cartilage tissue engineering applications. Scanning electron microscopic analysis indicated that the scaffolds provided favourable porous microstructures. In vitro degradation test showed that PGS/ PCL scaffolds got longer degradation time with surface degradation nature. PGS/PCL scaffolds seeded with Bone marrow-derived mesenchymal stem cells (BMSCs) and articular chondrocytes (ACCs) were cultured in vitro under the same condition. Short-term in vitro experiments confirmed that both seed cells could adhere and proliferate on the scaffold. Chondrogenic culture for cell-scaffold constructs confirmed BMSCs could differentiate into chondrocyte-like cells and form cartilage-specific matrix in PGS/PCL scaffolds. With cytocompatibility and biodegradation profile, PGS/PCL scaffolds get great potential for cartilage tissue engineering.

Salidroside-Mitigated Inflammatory Injury of Hepatocytes with Non-Alcoholic Fatty Liver Disease via Inhibition TRPM2 Ion Channel Activation.

PURPOSE: Oxidative stress plays an important role in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). TRPM2 ion channel functions as a molecular sensor for oxidative stress. The aim of this study was to examine the protective effects of Salidroside, a powerful antioxidative plant, on TRPM2 in an established in vitro model of NAFLD. METHODS: NAFLD model was established by palmitic acid (PA) in hepatic L02 cell lines and was added to the media at a final concentration of 400 µM. Cells were used as normal group, PA group and PA receiving varied concentrations of Salidroside (75µg/mL, 150µg/mL, 300µg/mL). After treating 24 hrs, MTT assay was used to detect cell viability, and ALT level was measured using an appropriate kit assay. Intracellular lipid accumulation was observed by Oil red O staining. Cytosolic Ca2+ concentrations were evaluated by flow cytometer with Fluo-3/AM. Quantitative RT-PCR was used to measure the mRNA expression of TRPM2, IL-1ß and IL-6, and the protein expressions of TRPM2, p-CaMKII and autophagy (LC3B, p62) were determined using Western blot. RESULTS: Treatment with Salidroside effectively restored liver injury and alleviated lipid droplet deposition in a dose-dependent manner, which was associated with inhibition of TRPM2/Ca2+/CaMKII pathway. Additionally, autophagic clearance was enhanced by intervention with Salidroside in a dose-dependent manner. Further investigation indicated that Salidroside down-regulated the mRNA expression of IL-1ß and IL-6-pro-inflammatory cytokines. CONCLUSION: These results suggest that Salidroside could alleviate inflammatory injury and steatosis via autophagy activation mediated by downregulation of the TRPM2/Ca2+/CaMKII pathway. Targeting the TRPM2 ion channel is a novel treatment strategy for oxidative stress-induced liver in NAFLD.

The hepatocyte phase of Gd-EOB-DTPA-enhanced MRI in the evaluation of hepatic fibrosis and early liver cirrhosis in a rat model: an experimental study.

AIMS: To evaluate the hepatocyte phase of Gd-EOB-DTPA-enhanced MRI in the early diagnosis of hepatic fibrosis and cirrhosis and assessment of liver function in a rat model. MAIN METHODS: In 2 groups of SD rats, liver fibrosis was induced in experimental animals by repetitive carbon tetrachloride injections, while the control group received saline injections. Five experimental rats and 2 control rats were randomly selected at weeks 4, 8, 12. One week after carbon tetrachloride administration, MRI (FIRM T1WI) scan was performed. Gd-EOB-DTPA (0.08mL) was injected into the rat's tail vein and hepatocyte phase images were obtained after 20min. The pre-enhanced phase and hepatocyte phase signal intensities (SI) were measured, and the relative contrast enhancement index (RCEI) was calculated. ANOVA analysis (LSD) of RCEI values in controls (n=6), hepatic fibrosis (n=7), and histopathologically-determined early cirrhosis group (n=6) was performed. KEY FINDINGS: RECI values showed a decreasing trend in the control group, hepatic fibrosis and early cirrhosis groups (1.11±0.43, 0.96±0.22, and 0.57±0.33, respectively). While the difference between the control and early cirrhosis groups was statistically significant (p=0.013), there was no significant difference in the hepatic fibrosis group vs the control (p=0.416) and the hepatic fibrosis group vs the early cirrhosis group (p=0.054). SIGNIFICANCE: Hepatocyte phase RCEI values obtained with Gd-EOB-DTPA-enhanced MRI scan indicate liver injury in hepatic fibrosis and early cirrhosis. RCEI values are helpful for early diagnosis of liver cirrhosis.