Associations between one-carbon metabolism and valproic acid-induced liver dysfunction in epileptic patients.

Valproic acid (VPA) has been widely used as an antiepileptic drug for decades. Although VPA is effective and well-tolerated, long-term VPA treatment is usually associated with hepatotoxicity. However, the underlying mechanisms of VPA-caused hepatotoxicity remain unclear. In this study, a total of 157 pediatric patients with epilepsy were recruited and divided into normal liver function (NLF, 112 subjects) group and abnormal liver function (ABLF, 45 subjects) group. We observed that MTHFR A1298C and MTHFR C677T variants may be linked to VPA-induced liver dysfunction (p = 0.001; p = 0.023, respectively). We also found that the MTHFR A1298C polymorphism was associated with a higher serum Hcy level (p = 0.001) and a lower FA level (p = 0.001). Moreover, the serum Hcy levels was strongly correlated with the GSH and TBARS concentrations (r = -0.6065, P < 0.001; r = 0.6564, P < 0.001, respectively). Furthermore, logistic analysis indicated that MTHFR A1298C/C677T polymorphisms and increased Hcy concentrations may be risk factors for VPA-induced liver dysfunction. These results suggested that individual susceptibility to VPA-induced liver dysfunction may result from MTHFR A1298C/C677T polymorphisms and increased Hcy levels. This study may be helpful for the prevention and guidance of VPA-induced liver dysfunction.

Diosgenin inhibits Wnt/ß-catenin pathway to regulate the proliferation and differentiation of MG-63 cells.

Dioscorea opposita Thunb has the effect of anti-osteoporosis, but whether its active ingredient diosgenin (DIO) has an anti-osteoporosis effect is unknown. The purpose of this study is to investigate the effect of DIO on the proliferation and differentiation of MG-63 cells. MG-63 cells were treated with different concentrations of DIO (0.001, 0.01, 0.1 and 1 µM) or 20 mM Wnt/ß-catenin signaling agonist-LiCl, and then their cell cycle and viability were analyzed by flow cytometry and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), respectively. To investigate osteoblast differentiation, alizarin red staining and ultraviolet spectrophotometer were used to determine the number of calcified nodules and the activity of alkaline phosphatase (ALP), respectively. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting were used to detect the expressions of proliferation-related, osteogenic-related and Wnt/ß-catenin signal pathway-related factors. After the cells were treated with low-concentration (0.001 or 0.01 µM) DIO, cell viability was significantly increased and the proportion of cells in S phase was increased. In addition, low-concentration DIO could significantly increase the expression of Ki67, proliferating cell nuclear antigen (PCNA), osteopontin (OPN), and osteocalcin (BGP), promote osteoblast differentiation, and suppress the expression of ß-catenin, Runx2 and cyclinD1. However, high concentrations of DIO showed the opposite effect. Low-concentration DIO obviously reversed the effect of LiCl on decreasing the number of calcified nodules and inhibiting the expression of OPN and BGP in cells. Low-concentration DIO might promote the proliferation and differentiation of MG-63 cell by inhibiting the Wnt/ß-catenin signal pathway.

Relationship between changes in the course of COVID-19 and ratio of neutrophils-to-lymphocytes and related parameters in patients with severe vs. common disease.

To assess the relationship between the neutrophil-to-lymphocyte ratio (NLR) and related parameters to the severity of coronavirus disease 2019 (COVID-19) symptoms. Clinical data from 38 COVID-19 patients who were diagnosed, treated and discharged from the Qishan Hospital in Yantai over the period from January to February 2020 were analysed. NLR and procalcitonin (PCT) were determined in the first and fourth weeks after their admission, along with the clinical characteristics and laboratory test results of these patients. Based on results as obtained on the first and fourth weeks after admission, five indices consisting of NLR, white blood cells, neutrophils, lymphocytes (LY) and monocytes (MON) were selected to generate receiver operating characteristic curves, while optimal cutoff values, sensitivities and specificities were obtained according to the Yuden index. Statistically significant differences in neutrophils, LY and the NLR were present in the severe vs. moderate COVID-19 group from the first to the fourth week of their hospitalisation. The cut-off value of NLR for predicting the severity of COVID-19 was 4.425, with a sensitivity of 0.855 and a specificity of 0.979. A statistically significant positive correlation was present between PCT and NLR in the severe group as determined within the first week of admission. NLR can serve as a predictor of COVID-19 disease severity as patients' progress from the first to the fourth week of their hospitalisation. The statistically significant positive correlation between levels of NLR and PCT in severe patients indicated that increases in NLR were accompanied with gradual increases in PCT.

Expression of miR-376 in blood of pregnant women with preeclampsia and its effect on 25-hydroxyvitamin D.

The study aims to investigate the clinical significance of regulating the expression of 25-hydroxyvitamin D (25-OH-VD) via microRNA (miRNA)-376c in the occurrence and development of preeclampsia (PE) in pregnant women. Peripheral blood and placental tissues were collected from pregnant women in 4 groups, including 67 normal pregnant women, 41 pregnant women with gestational hypertension, 40 pregnant women with mild PE and 51 pregnant women with severe PE. The expression of 25-OH-VD and miRNA-376c in peripheral blood were analyzed via reverse transcription-quantitative polymerase chain reaction (RT-qPCR); the protein expression of 25-OH-VD was analyzed via western blotting, and the clinical significance of its expression was also analyzed. The expression of miRNA-376c in peripheral blood in pregnant women was decreased (P<0.01), and the expression of 25-OH-VD in peripheral blood was significantly decreased (P<0.01); there was a significantly positive correlation between the expression of miRNA-376c and 25-OH-VD (P<0.01). There was a significantly positive correlation between miRNA-376c and the protein expression of 25-OH-VD in placental tissues (P<0.01). The downregulation of miRNA-376c expression in peripheral blood and placental tissues in pregnant women had significantly positive correlations with gestational age, plasma albumin level and fetal weight, but had significantly negative correlations with blood pressure and urinary protein level. (P<0.01). The downregulation of 25-OH-VD expression in placental tissues also had such correlations. The low expression of miRNA-376c in PE patients is involved in the occurrence and development of PE through downregulating the expression of 25-OH-VD.

Targeting STAT3 induces apoptosis and suppresses cell growth and invasion by inactivation of Slug signaling in retinoblastoma.

OBJECTIVE: Abnormalities in the STAT3 pathway are involved in the oncogenesis of several cancers. However, the mechanism by which dysregulated STAT3 signaling inhibited the progression of human retinoblastoma (RB) has not been elucidated. To investigate the role of targeting STAT3 in RB progression, we inhibited STAT3 with Y79 cells and depleted STAT3 with a siRNA. RESULTS: Our results demonstrate that targeting STAT3 inhibited survival and induced apoptosis of Y79 cells through downregulation of Slug and upregulation of PUMA. In addition, targeting STAT3 inhibited invasion and migration of Y79 cells through downregulation of Slug and upregulation of E-cadherin. Furthermore, Slug overexpression inhibited PUMA and E-cadherin upregulation in the Y79 cells with targeting STAT3. Targeting PUMA and E-cadherin reversed the role of targeting STAT3 in the Y79 cells. CONCLUSIONS: our findings showed that targeting STAT3 inhibited survival, invasion, and migration in Y79 cells through inactivation of Slug, resulting in the upregulation of PUMA and E-cadherin, and provide novel evidence that the STAT3/Slug pathway may be a new potential target for therapy of RB.

Clusterin (apolipoprotein J) facilitates NF-κB and Bax degradation and prevents I/R injury in heart transplantation.

BACKGROUND AND AIM: Ischemia-reperfusion (I/R) injury is an unavoidable event occurring during heart transplantation and is a key factor in graft failure and long-term survival rate of recipients. Therefore, there is an urgent need for the development of new therapies to prevent I/R injury. Clusterin is a hetero-dimeric glycoprotein with an antiapoptotic function. In this study, we investigated whether clusterin was cardioprotective in heart transplantation against I/R injury using an in vivo rat model and an in vitro cell culture system and we examined the underlying mechanisms of I/R injury. METHODS: Heart grafts from wild-type C57BL/6 mice were preserved in UW solution (control) or UW solution containing recombinant human apolipoprotein J (hr clusterin) for 24 hours. The preserved hearts were implanted into recipient mice of the same strain as the donors for 72 hours. The heart grafts were then taken for histopathological and gene expression analyses. An in vitro ischemia reperfusion model using H9C2 cells or H9C2/clusterin cDNA cells was constructed. The expression of clusterin, p65, Bax, Bcl-xL, IL-1ß, and TNF-α protein and mRNA in heart tissue and H9C2 cells was detected by Western blot, reverse transcription-polymerase chain reaction (RT-PCR), and quantitative RT-PCR assays. IL-1ß and TNF-α protein was detected by enzyme-linked immunosorbent assays. NF-kB activity was detected by an electrophoretic mobility shift assay and cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and flow cytometric analyses. RESULTS: Cold I/R caused severe morphologic myocardial injury to heart grafts from wild-type C57BL/6 mice whereas grafts from hr clusterin preservation showed less damage, as demonstrated by decreased cell apoptosis/death, decreased neutrophil infiltration, and the preservation of the normal structure of the heart. Clusterin reduced expression of p65, pre-inflammatory IL-1ß, TNF-α, and the pro-apoptotic gene Bax while it enhanced expression of the anti-apoptotic gene Bcl-xL in vitro and in vivo. Clusterin inhibited cell apoptosis/death and reduced pre-inflammation. CONCLUSION: Clusterin is a promising target for preventing cold I/R injury in heart transplantation. This study also shows that the resultant protective effects of clusterin are mediated by NF-κB signaling and Bax/Bcl-xL expression.

Clinical significance of serum 14-3-3 beta in patients with hepatocellular carcinoma.

OBJECTIVE: The 14-3-3 family of conserved regulatory proteins comprises the isoforms beta (ß), gamma (γ), zeta (ζ), sigma (ε), tau (η), and delta (σ), which are overexpressed and associated with a high risk of metastasis and poor survival in hepatocellular carcinoma (HCC). In the present study, we investigated whether serum 14-3-3 isoforms are related to HCC progression and patient survival. METHODS: Serum samples from 63 HCC patients who underwent surgical reSection 104 HCC patients who received non-surgical anti-HCC treatments, 50 patients with liver cirrhosis alone, 45 patients with chronic hepatitis alone, and 50 healthy subjects were collected between January 2006 and December 2010. Serum levels of 14-3-3 (ß, ε, γ, σ, and ζ) isoforms were measured by ELISA. The correlation between 14-3-3 (ß and σ) isoforms and clinicopathological factors was examined by logistic regression analysis. The feasibility of serum 14-3-3 ß for discriminating HCC patients was assessed by ROC curve analysis. Patient survival analyses were performed by Kaplan-Meier analyses and Cox regression models. RESULTS: Serum levels of 14-3-3 (ß and σ) were significantly higher in HCC patients than in those with liver cirrhosis, chronic hepatitis, and healthy subjects (p< 0.05). There was no difference in the serum levels of 14-3-3 ε, γ, and ζ between HCC and the other groups (p> 0.05). High levels of serum 14-3-3 ß were associated with vascular invasion (p= 0.016), TNM stage (p= 0.012), BCLC stage (p= 0.01), and early recurrence (p= 0.013). Patients with high levels of serum 14-3-3 ß had a poor prognosis. There was no significant association between 14-3-3 σ levels and clinicopathological parameters. A significant independent association between serum 14-3-3 ß and HCC was observed by univariate and multivariate analysis (p< 0.05). Serum 14-3-3 ß could effectively discriminate HCC patients at a cut-off point of 18.7 ng/mL, with 91.4% sensitivity and 75.3% specificity. CONCLUSIONS: Serum 14-3-3 ß is a potential biomarker for the diagnosis of early-stage HCC, and high levels of serum 14-3-3 ß were associated with metastasis and poor prognosis in HCC.

Genetic characteristics of the couple with non-syndromic sensorineural hearing loss and fertility guidance.

PURPOSE: We aim to report a genetic testing and fertility guidance for the deaf through analyzing pedigree and molecular genetic characteristics of the couple who have non-syndromic sensorineural hearing loss (NSHL). METHODS: One of hospitalized congenial deaf couple and family members were included in this study. The wife was twin pregnant woman and her gestational age was 31(+5) pregnant weeks. The DNA was extracted from peripheral blood and umbilical vein blood, respectively. Mutation screening of common deafness genes was performed in pregnant women and other family members. Nine common mutations in four major deafness genes, GJB2 (35delG, 176del16, 235delC, 299delAT), GjB3 (C538T), SLC26A4 (IVS7-2A>G, A2168G) and Mitochondrial 12S rRNA (A1555G, C1494T), were detected simultaneously with a microarray based method. SLC26A4 whole genome sequencing was carried out for the results of the DNA microarray. According to the test results, the couple chose abortion termination of pregnancy twins, and after one year obtained singleton pregnancy by artificial insemination by donor (AID). In week 16 of pregnancy, amniocentesis had been done to collect fetal somatic cell and extract DNA, and then the above tests had been repeated. RESULTS: The couple had SLC26A4 combined heterozygous mutation. Both parents had SLC26A4 single heterozygous mutation. Twin fetuses had SLC26A4 combined heterozygous mutation. The probability of naturally being pregnant and bearing deaf children for the pregnant women was 100%. Fetus obtained by AID had SLC26A4 single heterozygous mutation. After the birth of the baby, her hearing has been normal. CONCLUSIONS: To reduce children with congenital deafness, screening high mutation sites by microarray, combined with pedigree analysis and gene sequencing is effective, and should be used as a routine inspection item for the deaf before marriage and pregnancy. On the basis of genetic testing for the couple with hearing loss, human assisted reproductive technology is a viable option to avoid the birth of infant with hereditary deafness.

Increased serum mannose binding lectin levels are associated with diabetic retinopathy.

BACKGROUND: Mannose-binding lectin (MBL) may be implicated in the pathogenesis of diabetic retinopathy (DR) complications. We investigated serum MBL levels in type 2 diabetic patients with and without DR. METHOD: Serum MBL levels were determined in 184 type 2 diabetic patients with DR and 189 type 2 diabetic patients without DR matched for age, sex, and duration of diabetes. Multivariate analyses were performed using logistic regression models. The diagnostic value of MBL was compared with the HbA1c, Hs-CRP and with other known markers. RESULTS: We found that serum MBL levels were significantly higher in diabetes with DR as compared to without-DR [3456 (IQR, 3128-3800) ug/l and 2432 (IQR, 2100-2670) ug/l, respectively; P<0.0001]. In multivariate logistic regression analysis, after adjusting for all other significant factors, MBL remained can be seen as an independent DR marker with an adjusted OR of 1.002 (95% CI, 1.001-1.003; P<0.0001). Based on the ROC curve, the optimal cutoff value of serum MBL levels as an indicator for diagnosis of DR was projected to be 3050 ug/L, which yielded a sensitivity of 82.5% and a specificity of 88.0%, with the area under the curve at 0.907 (95% CI, 0.876-0.938). CONCLUSION: In type 2 diabetic patients, evaluated serum levels of MBL can be seen as an independent marker of DR even after correcting for possible confounding factors.