A Cysteinyl-tRNA Synthetase Mutation Causes Novel Autosomal-Dominant Inheritance of a Parkinsonism/Spinocerebellar-Ataxia Complex.

This study aimed to identify possible pathogenic genes in a 90-member family with a rare combination of multiple neurodegenerative disease phenotypes, which has not been depicted by the known neurodegenerative disease. We performed physical and neurological examinations with International Rating Scales to assess signs of ataxia, Parkinsonism, and cognitive function, as well as brain magnetic resonance imaging scans with seven sequences. We searched for co-segregations of abnormal repeat-expansion loci, pathogenic variants in known spinocerebellar ataxia-related genes, and novel rare mutations via whole-genome sequencing and linkage analysis. A rare co-segregating missense mutation in the CARS gene was validated by Sanger sequencing and the aminoacylation activity of mutant CARS was measured by spectrophotometric assay. This pedigree presented novel late-onset core characteristics including cerebellar ataxia, Parkinsonism, and pyramidal signs in all nine affected members. Brain magnetic resonance imaging showed cerebellar/pons atrophy, pontine-midline linear hyperintensity, decreased rCBF in the bilateral basal ganglia and cerebellar dentate nucleus, and hypo-intensities of the cerebellar dentate nuclei, basal ganglia, mesencephalic red nuclei, and substantia nigra, all of which suggested neurodegeneration. Whole-genome sequencing identified a novel pathogenic heterozygous mutation (E795V) in the CARS gene, meanwhile, exhibited none of the known repeat-expansions or point mutations in pathogenic genes. Remarkably, this CARS mutation causes a 20% decrease in aminoacylation activity to charge tRNACys with L-cysteine in protein synthesis compared with that of the wild type. All family members carrying a heterozygous mutation CARS (E795V) had the same clinical manifestations and neuropathological changes of Parkinsonism and spinocerebellar-ataxia. These findings identify novel pathogenesis of Parkinsonism-spinocerebellar ataxia and provide insights into its genetic architecture.

Increased cysteinyl-tRNA synthetase drives neuroinflammation in Alzheimer's disease.

BACKGROUND: Microglia-mediated neuroinflammation in Alzheimer's disease (AD) is not only a response to pathophysiological events, but also plays a causative role in neurodegeneration. Cytoplasmic cysteinyl-tRNA synthetase (CARS) is considered to be a stimulant for immune responses to diseases; however, it remains unknown whether CARS is involved in the pathogenesis of AD. METHODS: Postmortem human temporal cortical tissues at different Braak stages and AD patient-derived serum samples were used to investigate the changes of CARS levels in AD by immunocytochemical staining, real-time PCR, western blotting and ELISA. After that, C57BL/6J and APP/PS1 transgenic mice and BV-2 cell line were used to explore the role of CARS protein in memory and neuroinflammation, as well as the underlying mechanisms. Finally, the associations of morphological features among CARS protein, microglia and dense-core plaques were examined by immunocytochemical staining. RESULTS: A positive correlation was found between aging and the intensity of CARS immunoreactivity in the temporal cortex. Both protein and mRNA levels of CARS were increased in the temporal cortex of AD patients. Immunocytochemical staining revealed increased CARS immunoreactivity in neurons of the temporal cortex in AD patients. Moreover, overexpression of CARS in hippocampal neurons induced and aggravated cognitive dysfunction in C57BL/6J and APP/PS1 mice, respectively, accompanied by activation of microglia and the TLR2/MyD88 signaling pathway as well as upregulation of proinflammatory cytokines. In vitro experiments showed that CARS treatment facilitated the production of proinflammatory cytokines and the activation of the TLR2/MyD88 signaling pathway of BV-2 cells. The accumulation of CARS protein occurred within dense-core Aß plaques accompanied by recruitment of ameboid microglia. Significant upregulation of TLR2/MyD88 proteins was also observed in the temporal cortex of AD. CONCLUSIONS: The findings suggest that the neuronal CARS drives neuroinflammation and induces memory deficits, which might be involved in the pathogenesis of AD.

Characteristics and clinical significance of polyploid giant cancer cells in laryngeal carcinoma.

OBJECTIVES: We aimed to construct an induction system for polyploid giant cancer cells (PGCCs), as well as to investigate PGCC features and clinical significance. METHODS: A laryngeal neoplasm-PGCC induction system was constructed using paclitaxel liposomes (PTX). We used western blots to compare expression of epithelial-mesenchymal transition-related proteins, stem cell interrelated proteins, and cyclin-associated proteins. We then measured PGCC count in tissue samples of patients with laryngeal neoplasms and analyzed its relationship with prognosis. Statistical significance was determined using t-tests. RESULTS: PTX successfully induced PGCCs. Western blotting showed that CyclinB1, CDC25C, CDK1, E-cadherin, and EIF-4A expression decreased in PGCCs compared with normal cancer cells, whereas vimentin and CD133 expression increased. Number of PGCCs in laryngeal cancer tissues and overall survival time were inversely correlated (P < .05). CONCLUSIONS: PTX successfully induces PGCC formation in laryngeal carcinoma, which may be the cause of poor prognosis in patients with laryngeal cancer.Level of Evidence: 4.

Serum-derived exosomes promote CD8+ T cells to overexpress PD-1, affecting the prognosis of hypopharyngeal carcinoma.

BACKGROUND: Hypopharyngeal cancer (HPC) is associated with a poor prognosis and a high recurrence rate. Immune escape is one of the reasons for the poor prognosis of malignant tumors. Programmed cell death ligand 1 (PD-L1) and programmed cell death-1 (PD-1) have been shown to play important roles in immune escape. However, the role of PD-1/PD-L1 in HPC remains unclear. In this experiment, we investigated the effect of exosomes from HPC patient serum on CD8+ T cell function and PD-1/PD-L1 expression and, thus, on prognosis. We hope to provide guidance for the identification of new targets for HPC immunotherapy. METHODS: PD-1 and CD8 expression in 71 HPC tissues and 16 paracarcinoma tissues was detected by immunohistochemistry. Concurrently, the clinicopathological data of the patients were obtained to conduct correlation analysis. Exosomes were isolated from serum and then identified by Western blotting (WB), transmission electron microscopy (TEM), and nanoparticle tracking analysis (NTA). Flow cytometry was used to assess the activity of CD8+ T cells after exosome stimulation. The effects of exosomes on the ability of CD8+ T cells to kill FaDu cells were assessed by CCK-8 assay. The expression of IL-10 and TGF-ß1 was measured by enzyme-linked immunosorbent assay (ELISA). PD-L1 expression in HPC tissue samples was evaluated by immunohistochemistry, and the relationship between PD-1/PD-L1 expression and prognosis was investigated with patient specimens. RESULTS: PD-1 expression was significantly upregulated on CD8+ T cells in tumor tissues compared with those in normal tissues. The overall survival (OS) and disease-free survival (DFS) of PD-1-overexpressing patients were decreased. Serum exosomes from patients can elevate PD-1 expression on CD8+ T cells and suppress their killing capacity and secretory function. The rate of positive PD-L1 expression was increased in HPC tissues compared with paracancerous tissues. The DFS and OS of the PD-1(+)-PD-L1(+) group were significantly lower than those of the PD-1(-)-PD-L1(-) group. CONCLUSION: Our findings indicate that serum exosomes from HPC patients can inhibit CD8+ T cell function and that the PD-1-PD-L1 pathway plays an important role in the immune escape of HPC. Exosomes combined with immunotherapy may guide the treatment of patients with advanced disease in the future.

PEMT G523A (V175M) is associated with sporadic Alzheimer's disease in a Chinese population.

There is evidence that increased concentrations of circulating homocysteine are associated with Alzheimer's disease (AD). Phosphatidylethanolamine N-methyltransferase (PEMT) is an important catalyst involved in the production of homocysteine. We investigated the association of a functional single nucleotide polymorphism (rs7946) in PEMT with sporadic AD risk in a Han Chinese population that included 386 AD patients and 366 controls. PEMT G523A was genotyped by either sequencing or PCR-restriction fragment length polymorphism analysis. The plasma homocysteine concentrations of 210 subjects were determined by high-performance liquid chromatography. Significant higher frequency of the A allele was detected in AD cases than in controls (A vs. G, p = 0.007, OR = 1.482, 95% CI 1.114-1.972). After adjusting for gender, age/age at onset, and APOE ε4 status, logistic analysis showed rs7946 was associated with AD in a dominant model (AA + GA vs. GG, p = 0.007, OR = 1.596, 95% CI 1.138-2.240). When stratified by APOE ε4 status or gender, the significant difference was only observed in the APOE ε4 non-carriers and in the female subjects, respectively. We did not find a relationship of this polymorphism with plasma homocysteine levels. These results suggested that PEMT G523A is associated with AD and that the A allele is an APOE ε4-independent risk factor for AD among Han Chinese women.

Association analysis of CbetaS 844ins68 and MTHFD1 G1958A polymorphisms with Alzheimer's disease in Chinese.

Folate deficiency and elevated plasma homocysteine play important roles in pathogenesis of Alzheimer's disease (AD). The aim of this study was to test the association of folate metabolism-related genes, cystathionine beta-synthase gene (CbetaS) and 5, 10-methylenetetrahydrofolate dehydrogenase gene (MTHFD1), with sporadic AD. The CbetaS 844ins68 polymorphism was determined by PCR and the MTHFD1 G1958A single nucleotide polymorphism (rs2236225) by PCR-RFLP. No significant difference of allele and genotype contributions of the CbetaS polymorphism between AD cases and controls was detected, before and after stratification by APOE epsilon4-carrying status, age/age at onset and genders. No significant difference of allele and genotype contributions of the MTHFD1 polymorphism between AD cases and controls was detected in total samples. When stratified by age/at onset age, we found that A allele and AA genotype frequencies in cases were higher than in controls and the differences were close to significant [A vs. G, P = 0.032, Odds ratio (OR) 1.642, 95% CI 1.040-2.591; AA + GA vs. GG, P = 0.068, OR 1.665, 95% CI 0.961-2.885; AA vs. GG, P = 0.059, OR 3.458, 95% CI 0.894-13.369] in <65 years groups, which suggested that the MTHFD1 G1958A A allele might be a weak risk factor for early onset AD although it needs further confirmation.

[MRI features of olfactory bulb volume in healthy middle and old-aged persons.].

OBJECTIVE: To investigate the differences of olfactory bulb (OB) volumes between younger and older, male and female, left-side and right-side in healthy middle and old-aged persons by MRI. METHODS: Ninety five healthy middle and old-aged volunteers (male:female = 45:50) were divided into 2 groups, group one included persons aged from 50 to 69, group two included persons elder than 70. The left-side, right-side and both-side volumes of OB, the volumes of brain and the ratio of OB/brain were measured by MRI. RESULTS: (1) The left-side and both-side volumes of OB (x(-) +/- s), the volumes of brain [(39.89 +/- 8.7) mm(3), (81.70 +/- 16.8) mm(3) and (1281.86 +/- 140.2) cm(3)] in 50 - 69 years old group were respectively larger than those in >/= 70 years old group [(34.45 +/- 10.4) mm(3), (72.10 +/- 19.3) mm(3) and (1165.77 +/- 165.3) cm(3)], and the differences reached statistical significance (t were respectively 2.649, 2.449, 3.516, all P < 0.05). There were no significant differences of right-side OB volumes and the ratio of OB/brain between 50 - 69 years old group and >/= 70 years old group (t were respectively 1.904, 0.616, each P > 0.05). (2) The male's OB volumes of left-side, right-side and both-side, the brain volumes and the ratio of OB/brain were respectively larger than females', and the differences reached statistical significance (t were respectively 4.461, 3.630, 4.399, 3.800, 2.400, all P < 0.05). (3) The right-side OB volumes were larger than left-side's and significant differences were found in female group, 50 - 60 years old group and >/= 70 years old group (t were respectively 2.732, 2.117, 3.516, all P < 0.05). There were no significant differences of OB volumes between left-side and right-side in female (t = 2.649, P = 0.110). The ratio of right-side OB/brain were larger than the ratio of left-side's and the differences reached statistical significance (t = 3.183, P = 0.002). CONCLUSIONS: MRI could be used to measure the volume of OB. The older the people, the smaller the OB volumes. There was no influence of age on the ratio OB/brain. The OB volumes of right-side were larger than those of left-side. The OB volumes of male were larger than those of female.

[The application of Montreal cognitive assessment in urban Chinese residents of Beijing].

OBJECTIVE: To study the distribution of Montreal cognitive assessment (MoCA) scores in terms of age and educational level in Chinese residents aged 50 years and over and to offer a benchmark for a cutoff score. METHODS: A total of 281 residents aged 50 years or older was drawn randomly in the urban areas of Beijing, including 215 healthy elderly controls (NC) and 66 patients meeting the clinical criteria of mild cognitive impairment (MCI). The final scores for MoCA were given in the form of mean percentage distributions specific for age, sex and educational level so as to compare the validity of MMSE mini-mental state examination and MoCA in detecting MCI. By a fitting multiple regression model the influence of the factors on MMSE and MoCA was assessed. RESULTS: Using a cutoff score of 26, MMSE had a sensitivity of 24.2% to detect MCI, whereas MoCA detected 92.4% of the MCI subjects. We found a bivariate correlation between MoCA scores and both the factors of age and educational level (P < 0.001). CONCLUSIONS: MoCA is a brief cognitive screening tool with high sensitivity and specificity for detecting MCI as currently conceptualized in patients performing normally on MMSE. Our adjustment in the cutoff scores would improve the detection of MCI and Alzheimer's disease by reducing the number of false negatives. MoCA scores should be used to identify current cognitive difficulties but not to make formal diagnoses.

Evidence of a relationship between infant birth weight and later diabetes and impaired glucose regulation in a Chinese population.

OBJECTIVE: The aim of this study was to determine the influence of birth weight, a marker of fetal growth, on the development of later impaired glucose metabolism throughout the life span of people living in China. RESEARCH DESIGN AND METHODS: We recorded detailed anthropometric data including height, weight, and health status and measured blood glucose levels and insulin concentrations after fasting and at 120 min of a standard oral glucose tolerance test from 2,019 eligible subjects born between 1921 and 1954 to investigate the risk of developing type 2 diabetes and impaired glucose regulation (IGR). RESULTS: The diabetes and IGR groups were characterized by significantly lower birth weight (P < 0.001), smaller head circumference (P < 0.001), smaller ponderal index (P = 0.007), and shorter length (P = 0.004) compared with those in the normal glucose tolerance group. Using multiple logistic regression analysis, we observed that birth weight remained significantly associated with diabetes and IGR after adjustments for possible confounding variables at birth and in adult life such as sex, age, central obesity, smoking status, alcohol consumption, dyslipidemia, family history of diabetes, and occupational status (P = 0.027). There was a significantly increased risk of getting diabetes and IGR for those with low birth weight (odds ratio 1.748 [95% CI 1.018-3.001], P = 0.043). CONCLUSIONS: The results confirm that lower birth weight is an independent risk factor for later diabetes or IGR and show for the first time that this risk factor also applies for a Chinese population.

The Fas gene A-670G polymorphism is not associated with sporadic Alzheimer disease in a Chinese Han population.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by excessive neuronal loss, intracellular neurofibrillary tangles and extracellular deposition of amyloid beta-peptide (Abeta). The Fas antigen is a cell surface receptor-mediating cell apoptosis. Several lines of evidence have made Fas/Fas ligand induced apoptosis play an important role in the pathogenesis of AD. Moreover, the Fas gene is located on chromosome 10q24.1, a region of linkage to late-onset AD. Several reports have investigated the association between a single nucleotide polymorphism (SNP) that is located at position -670 of Fas gene and AD, but yielded ambiguous results. To figure out the association of this SNP with sporadic AD in Chinese Han population, we have analyzed 509 patients with AD and 561 controls for the genetic association studies. Our results indicate that the distribution of the Fas genotypes (chi(2) = 0.66, P = 0.72) and alleles (chi(2) = 0.70, P = 0.40) did not differ significantly. The similar results were observed when AD and control groups were stratified by age/age at onset and sex (P > 0.10). The present data revealed no significant effect of the genotypes on the age of onset for developing AD, and no significant association between the genotypes and the severity of the disease.

[Cigarette smoking and Parkinson's disease: a population based case-control study].

OBJECTIVE: To explore the association with cigarette smoking for Parkinson's disease (PD). METHODS: One hundred and fourteen PD cases and 205 controls matched on gender and race were recruited from ongoing PD prevalence survey and identified at the neurological clinic of Peking Union Medical College Hospital. Face to face questionnaire interview was carried out and data on smoking and alcohol consumption were analyzed in a population-based case control study. RESULTS: With never-smokers as the reference category, we observed reduced risk for PD among ever smokers (OR = 0.49, 95% CI: 0.30 - 0.79) current smokers (OR = 0.44, 95% CI: 0.23 - 0.86) and ex-smokers (OR = 0.54, 95% CI: 0.30 - 0.96). When comparing with non-smokers, the ever smokers stratified by years of smoking had an inverse association with those whose smoking history longer than 20 years (OR = 0.35, 95% CI: 0.18 - 0.70) and an mild protective association with those who smoked less than 20 years (OR = 0.61, 95% CI: 0.35 - 1.07). Those who had quitted smoking for more than 20 years were less likely to have the disease than never smokers, and those who had quitted for less than 20 years were least likely to have PD. Those current smokers were still least likely to have the disease. Significant inverse gradient with pack-day smoker (trend P < 0.05), and the inverse association for cigarette smoking and PD were found not bing confounded by alcohol consumption. CONCLUSION: The inverse association between PD and cigarette smoking and history of cessation was found. Further studies need to provide biochemical evidence on the relation between smoking and its protective effect on PD.

Parkinson's disease and smoking: an integral part of PD's etiological study.

OBJECTIVE: To explore the association of Parkinson's disease (PD) with cigarette smoking. METHODS: One hundred of fourteen PD patients were compared with 205 control subjects who were matched by gender, race and residency. A previously validated questionnaire including smoking, alcohol/tea consumption as well as some other environmental exposure data was administered. RESULTS: With never-smokers as the reference category, we observed reduced risk for PD among ever smokers (OR=0.49, 95% CI: 0.30 to 0.79) current smokers (OR=0.44, 95% CI: 0.23 to 0.86) and ex-smokers (OR=0.54, 95% CI: 0.30 to 0.96). When ever smokers were stratified by years of smoking, there was an inverse correlation between those whose smoking history was longer than 20 years (OR=0.40 95% CI: 0.21 to 0.81) and an even mild protective correlation between those who smoked less than 20 years (OR=0.57, 95% CI: 0.33 to 0.99). Those who had quitted smoking for more than 20 years were less likely to have the disease than never smokers, and those who had quitted for less than 20 years were least likely to have PD, while those who were current smokers were still least likely to have the disease. We found significant inverse gradient with pack-day smoking (trend P<0.05), and the inverse correlation between cigarette smoking and PD was not confounded by alcohol/tea consumption and other confounding bias. CONCLUSIONS: The inverse correlation between Parkinson's disease risk and smoking as well as the trend of gradient dose response is again observed in our study. More future researches are needed to confirm these correlations and to explore further biochemical evidence.

[Serum levels of macrophage colony stimulating factor in the patients with Alzheimer's disease].

OBJECTIVE: To investigate the availability of serum level of macrophage clony stimulating factor (M-CSF) as a marker for early diagnosis of Alzheimer's disease (AD). METHODS: The serum levels of M-CSF in 70 patients with AD, 52 healthy controls, 22 patients with VAD (vascular dementia) were measured and the serum levels of IL-1 beta, IL-6, TNF-alpha in 32 patients with AD and 20 controls were measured as well. RESULTS: Serum levels of M-CSF were significantly elevated in patients with AD when compared with healthy controls (P < 0.01) and VAD controls (P < 0.05) respectively. At the early stage of mild dementia and middle dementia, serum levels of M-CSF were significantly elevated, but at the later stage of severe dementia, they returned to normal level. Serum levels of IL-1 beta were significantly elevated in AD patients compared with controls (P < 0.05), and serum levels of TNF-alpha and IL-6 were within the normal range in patients with AD. CONCLUSIONS: The results suggest that serum M-CSF level may provide a convenient and sensitive means for the early diagnosis of Alzheimer's disease.