TBC1D24-related familial infantile multifocal myoclonus: Description of a new Chinese pedigree with a 20 year follow up.

INTRODUCTION: Disorders associated with mutations in the Tre2/Bub2/Cdc16 (TBC)1 domain family member 24 gene (TBC1D24) present a wide range of phenotypes, ranging from mild to fatal seizure diseases, non-syndromic deafness, and complex syndromes such as deafness, onychodystrophy, osteodystrophy, and mental retardation(DOOR syndrome). In this study, we introduce three siblings of a previously unreported Chinese family with familial infantile myoclonic epilepsy caused by a homozygous TBC1D24 mutation. METHODS: Genomic DNA was extracted from whole blood of the proband, his parents, and sisters. TBC1D24 exomes were sequenced by whole exome sequencing then analyzed by genetic analysis with Sanger sequencing validation. The patients were followed up for more than 20 years to summarize their clinical features. RESULTS: Genetic analysis identified a homozygous TBC1D24 mutation (c.241_252del12) in the proband and his sisters. Prediction models suggest that the mutation leads to an alteration in the properties and structure of the TBC1D24 protein, especially in the folding direction of the loop region, which is likely to decrease protein activity. The patients manifested with early-onset myoclonic epilepsy, were prone to status epilepticus, and seizures only occurred during wakefulness. Imaging characteristics included cerebellar atrophy and abnormal cerebellar signals. CONCLUSION: We report a pedigree case of infantile myoclonic epilepsy caused by a homozygous TBC1D24 mutation. Our long-term clinical follow-up not only enriches the clinical phenotype of the disease, but also provides a clinical experience for the early diagnosis and clinical treatment of the disease.

Tripartite motif protein 25 is associated with epirubicin resistance in hepatocellular carcinoma cells via regulating PTEN/AKT pathway.

Tripartite motif protein 25 (TRIM25) expression was altered in various human cancers. Herein, we found that the expression of TRIM25 was elevated in hepatocellular carcinoma (HCC) tissues and cell lines. Knockdown of TRIM25 increased the sensitivity of HCC HepG2 cells to epirubicin (EPI), as indicated by reduced cell viability, enhanced cell apoptosis, and downregulated P-glycoprotein (P-gp) and multiple drug-resistance protein 1 (MRP1). Moreover, TRIM25 knockdown strengthened the effects of EPI on phosphatase and tensin homolog (PTEN) and phosphorylated (p)-AKT. However, overexpression of TRIM25 exerted an opposite effect, weakening the sensitivity of Huh7 to EPI, and obviously increasing PTEN and reducing p-AKT. Most important, all the changes induced by TRIM25 overexpression in Huh7 were reversed with additional treatment of LY294002 (an AKT pathway inhibitor). Notably, coimmunoprecipitation experiments confirmed the interaction between TRIM25 and PTEN. Knockdown of TRIM25 resulted in reduced ubiquitination of PTEN protein. Collectively, our data suggested that TRIM25 enhanced EPI resistance via modulating PTEN/AKT pathway, and targeting TRIM25 may enhance the sensitivity of HCC cells toward chemotherapy drugs.

Protective effect of pantoprazole against sepsis-induced acute lung and kidney injury in rats.

To investigate the effect of pantoprazole on acute lung and kidney injury with sepsis and its possible mechanism. Rats were randomly divided into six groups, the status and lung wet/dry weight ratio were determined at various time points. Hematoxylin Eosin staining (HE) for pathological changes in the lungs and kidneys of rats with sepsis. Western blot (WB) and immunohistochemistry were used to detect the pulmonary surfactant protein A(SPA) and D(SPD). The levels of markers for kidney damage in serum and urine various time points were measured by ELISA. The apoptosis in lung and kidney tissue of rats were detected using TUNEL assay. Subsequently, the cell apoptosis of LPS-induced BEAS-2B and HK-2 cells after pantoprazole treatment were detected using flow cytometry. The levels of RHOA/ROCK signaling pathway proteins in the lung and kidney tissues and cells were detected using WB. Our results indicated that Pantoprazole could suppress the expression of inflammatory factors in the blood and alleviate pathological damage of lung and kidney tissues in rats with sepsis. Pantoprazole treatment could reduce apoptosis in lung and kidney tissues and inhibit cell apoptosis induced by LPS. In addition, pantoprazole can inhibit RHOA/ROCK signaling pathway proteins and the levels of inflammatory factors in LPS-induced BEAS-2B and HK-2 cells. Pantoprazole can improve the symptoms of acute lung and kidney injury in septic rats, which suggested that pantoprazole might be used to guide the treatment of sepsis.

[Clinical and variation analysis of three Chinese families affected with glutaric acidemia type 1].

OBJECTIVE: To detect potential variation in glutaryl-CoA dehydrogenase (GCDH) gene among three Chinese families affected with glutaric acidemia type Ⅰ(GA-1) and correlate the genotypes with phenotypes. METHODS: Genomic DNA was extracted from peripheral blood samples derived from three patients with GA-1 and their family members. The coding regions of the GCDH gene were amplified with PCR and subjected to Sanger sequencing. RESULTS: The clinical manifestation of the patients varied from macrocephaly to severe encephalopathy, with notable phenotypic difference between siblings carrying the same variation. In pedigrees 1 and 2, the probands have carried compound heterozygous variations c.1133C>T(p.Ala378Val) and c.1244-2A>C, which were derived their fathers and mothers, respectively. In pedigree 3, the proband has carried compound heterozygous variation c.339delT (p.Tyr113) and c.406G>T (p.Gly136Cys). Among these, variations c.339delT and c.1133C>T were verified as novel by retrieval of dsSNP, HGMD and 1000 genome database. Bioinformatic analysis suggested that above variations can affect protein function and are probably pathogenic. CONCLUSION: Above discovery has expanded the mutation spectrum of the GCDH gene. No correlation was found between the clinical phenotype and genotype of GA-1 patients.

[Clinical investigation and genetic analysis of a Chinese family with glutaric acidemia type I].

OBJECTIVE: To review the clinical features of a families affected with glutaric acidemia type I (GA-1) and screen potential mutations in glutaryl-CoA dehydrogenase (GCDH) gene. METHODS: Clinical data of the patients and their family members was analyzed. Genomic DNA was extracted from peripheral blood samples. The 11 exons and flanking sequences of the GCDH gene were amplified with PCR and subjected to direct DNA sequencing. RESULTS: Two patients have manifested macrocephaly. Imaging analysis revealed arachnoid cyst and subdural effusion. The elder sister had encephalopathy crisis. The younger sister had significantly raised glutaric acid, whilst the elder sister was normal during the non-acute phase. Genetic analysis has revealed a homozygous c.1244-2A> C mutation of the GCDH gene in both patients. CONCLUSION: The clinical features and mutation of the GCDH gene have been delineated in a Chinese family affected with GA-1. The c.1244-2A> C mutation may be particularly common in the Chinese population.

[The value of combination of the mortality in emergency department sepsis score and blood lactate level in the risk stratification of severe sepsis in the emergency department].

OBJECTIVE: To evaluate the combination of the mortality in emergency department sepsis (MEDS) score with blood lactate level in the risk stratification of patients with severe sepsis in the emergency department (ED). METHODS: 665 adult patients with severe sepsis admitted from May 2011 to December 2012 in ED were found to be eligible for the study. MEDS score, acute physiology and chronic health evaluation II (APACHEII) score, and arterial blood lactate was determined, and the outcomes in 28 days were recorded. Logistic regression analysis was used to evaluate the relationship between each predictive factor score and prognosis. Each predictive factor was compared with the areas under the receiver operating characteristics (ROC) curve (AUC). RESULTS: The mortality in 28 days was 34.6% in 665 patients. The mortality in group of MEDS score 12-27 was significantly higher than that group of MEDS score<12 [51.0% (156/306) vs. 20.6% (74/155), χ(2)=28.414, P=0.000]. In the meantime, APACHEII score and blood lactate level were also significantly higher in group of MEDS score 12-27 than those in group with MEDS score<12 [APACHEII score: 26.4±10.6 vs. 21.7±8.1, t=-3.555, P=0.002; lactate: 4.9 (2.3, 9.9) mmol/L vs. 3.9 (1.5, 8.9) mmol/L, Z=-2.352, P=0.023]. Kaplan-Meier survival analysis showed significant difference in the two groups (the Log Rank test 36.71, P<0.01). The levels of 3 predictive factors were predominantly higher in non-survivors than survivors [MEDS score: 14.1±6.7 vs. 8.2±4.5, t=-6.929, P=0.000; APACHEII score: 28.1±7.1 vs. 22.2±11.3, t=-6.472, P=0.000; lactate: 5.4 (2.9, 11.0) mmol/L vs. 3.8 (1.2, 9.1) mmol/L, t=-6.472, P=0.004]. The AUCs were 0.813, 0.706 and 0.727 for MEDS score, APACHEII score and blood lactate respectively. The predictive ability for 28-day mortality of MEDS score was better than blood lactate (P=0.008) and APACHEII score (P=0.005). The AUC of MEDS score combined with lactate was 0.865, and 28-day mortality prediction was better than MEDS score (AUC 0.865 vs. 0.813, P<0.001). The sensitivity (83.1%), specificity (93.2%), positive prediction value (PPV, 62.4%), and negative prediction value (NPV, 92.1%) for MEDS score combined with lactate were highest among all predictors. CONCLUSIONS: MEDS score combined with lactate is a good risk stratification tool for emergency patients with severe sepsis, and its prognostic capability is better than either MEDS score, APACHEII score or blood lactate.