[Identification and functional analysis of combined oxidative phosphorylation deficiency 28 gene mutation].

Objective: To report a case of combined oxidative phosphorylation deficiency 28 (COXPD28) in China, identified the pathogenic mutation and explored the pathogenic mechanism preliminarily. Methods: The clinical characteristics of a patient with COXPD28 were retrospectively analyzed and the pathogenic mutations were identified by mitochondrial gene sequencing and whole exome sequencing. The wild-type and mutant plasmids of pathogenic genes were constructed, and effect of mutation on protein expression by quantitative real-time PCR (qPCR) and Western blot were evaluated. Statistical methods mainly used one-way ANOVA and LSD test. Results: A 21 year old female patient presented with lactic acid poisoning due to repeated chest distress and wheezing since childhood. The sequencing of the whole exon group gene found that solute carrier family 25 member 26 (SLC25A26) gene had a compound heterozygous mutation (c.34G>C, p.A12P; c.197C>A, p.A66E), which was the first report in China. In vitro function test showed that the expression levels of SLC25A26 mRNA and S-adenosylmethionine carrier (SAMC) protein in cells transfected with SLC25A26 mutant plasmid were significantly lower than those transfected with wild type plasmid. The p.A66E mutant plasmid reduced the expression level of SLC25A26 mRNA and SAMC protein to 6% and 26% of wild type plasmids respectively (both P<0.001), while p.A12P mutant plasmid decreased to 62% and 82% of wild type plasmids respectively (P<0.001, P=0.044). When the double mutant (p.A66E+p.A12P) plasmids were co-transfected, the expression levels of SLC25A26 mRNA and SAMC protein decreased to 47% and 57% of the wild type plasmids, respectively (P<0.001, P=0.001). Conclusion: The pathogenic mutation gene of this patient with COXPD28 is SLC25A26 gene mutation (p.A66E, p.A12P), which causes the decrease of SLC25A26 expression level, mitochondrial oxidative phosphorylation dysfunction, and induces COXPD28.

Pineal parenchymal tumours: minimum apparent diffusion coefficient in prediction of tumour grading.

Retrospective assessment was performed of the minimum apparent diffusion coefficient (minADC) together with quantitative assessment of the histopathological features of resected tumour tissue and the Ki-67 labelling index (LI) for predicting pineal parenchymal tumour (PPT) grade. Magnetic resonance images of tumours from 26 male and female patients (mean age 27.7 years) with pathologically confirmed PPTs, comprising 10 pineocytomas (PCs), seven pineal parenchymal tumours of intermediate differentiation (PPTIDs) and nine pineoblastomas (PBs), were reviewed retrospectively. Using ADC maps derived from isotropic diffusion-weighted images, the minADC value of each tumour was determined postoperatively from several regions of interest defined in the tumour, excluding cystic, necrotic, haemorrhagic or calcified components. Surgical intervention was undertaken in all cases. The Ki-67 LI was measured in surgical specimens. Mean minADC and Ki-67 LI showed significant differences among PCs, PPTIDs and PBs, and minADC was negatively correlated with Ki-67 LI. It is concluded that the minADC of PPTs provide useful additional information when predicting tumour grading.

Value of diffusion-weighted imaging in grading tumours localized in the fourth ventricle region by visual and quantitative assessments.

This study investigated visual and quantitative assessment of diffusion-weighted imaging (DWI) for grading tumours localized in the fourth ventricle region. Patients were diagnosed histopathologically and classified into two groups: those with high-grade (World Health Organization [WHO] grades III and IV) and those with low-grade tumours (benign, WHO grades I and II). DWI signal intensity was described using a five-point scale. Minimum apparent diffusion coefficient (ADC) values were obtained from areas with the lowest signal. The mean signal intensity was significantly higher in high-grade than in low-grade tumours. The mean minimum ADC value was significantly lower in high-grade than low-grade tumours. Marked hyperintensity had sensitivity, specificity, positive predictive value and negative predictive value of 89.7%, 100%, 100% and 94.2%, respectively, when used as a diagnostic tool for high-grade tumours compared with 96.6%, 97.9%, 96.6% and 97.9%, respectively, when using a minimum ADC of 0.9 × 10(-3) mm(2)/s as a diagnostic marker. It was concluded that DWI is helpful in predicting the grades of tumours in the fourth ventricle region.

A unique magnetic resonance imaging feature of glioblastoma multiforme: the 'pseudopalisade' sign.

This study was designed to investigate the unique magnetic resonance imaging (MRI) appearance of histopathologically-proven glioblastoma multiforme (GBM) with pseudopalisade necrosis and to assess its value for grading gliomas and providing a differential diagnosis. The study included 169 patients with intracranial masses who underwent surgery and had a proven histopathological diagnosis: 50 with GBM, 77 with gliomas (46 grade II and 31 grade III) and 42 with other intracranial masses (20 metastases, 14 lymphomas and eight abscesses). All patients underwent preoperative brain MRI including post-contrast T(1)-weighted imaging. The presence of the 'pseudopalisade' sign on post-contrast T(1)-weighted images was compared among the different types of brain mass. The frequency of the 'pseudopalisade' sign in GBMs (94.00%) was significantly higher than that seen in grade II and III gliomas (11.69%) and other intracranial masses (7.14%). The 'pseudopalisade' sign on post-contrast T(1)-weighted images was useful for grading gliomas and for differentiating GBM from other brain masses.

Haemorrhage detection in brain metastases of lung cancer patients using magnetic resonance imaging.

Magnetic resonance susceptibility-weighted imaging (SWI) is a new, highly-sensitive technique used to detect haemorrhage. This study evaluated the ability of magnetic resonance imaging (MRI) to detect haemorrhage in 45 lung cancer patients with brain metastases and compared the results with T2*weighted imaging (T2*WI) and contrast-enhanced T1-weighted imaging (CE-T1WI). Eighty-nine haemorrhagic brain metastases were identified in 31 patients using SWI, 68 were identified in 23 patients using T2*WI and 46 were identified in 14 patients using CE-T1WI. Most micro-bleeds could only be identified by SWI. It was concluded that haemorrhage is a frequent occurrence in brain metastases originating from lung cancer and that haemorrhage can be detected using SWI in a majority of brain metastases patients.

Identification of cytochrome P450 2E1 in rat brain.

We have isolated cDNA fragments that were originated from P450 2E1 in rat brain by PCR analysis. Their size matched up to what we expected based on the reported sequence of rat liver P450 2E1 mRNA. Internal structure of the longest first-round PCR products were investigated by Southern blot analysis and "nested" PCR. Their results confirmed that PCR products actually originated from P450 2E1 mRNA in rat brain. RT-PCR was also carried out using P450 2E1 specific primers and the size of the product was exactly as we expected for P450 2E1. These experimental evidences should clarify the presence of P450 2E1 in rat brain.

Physiological indicators and susceptibility of plants to acidifying atmospheric pollution: a multivariate approach.

Plant susceptibility to acidifying atmospheric pollution was investigated for a wide range of plant species under normal environmental conditions. Physiological and biochemical parameters were recorded for each plant species. Multivariate analysis using hierarchical clustering, principal component analysis ordination, as well as regression analysis were used to interpret the data. Analysis of the data separated plants into pioneer tree species, ruderal herbaceous species and climax, slow growing species. Pioneer and ruderal species showed higher potential for foliar nitrate assimilation coupled with higher base cation content and a tendency to a more neutral leaf pH than climax species. Acidic buffering capacities for leaf tissue from pioneer and ruderal species were higher than those of climax species. Nitrate assimilation produces hydroxyl ions, which in conjunction with high base cation content and more neutral leaf pH, provides better metabolic buffering and therefore make pioneer and ruderal species less susceptible to acidifying inputs. Climax species are less able to buffer against acidifying inputs and are subsequently more prone to acidifying air pollution damage.

Molecular cloning and sequencing of sulfated glycoprotein-2 cDNA from testis of mouse: implications of two different mRNAs of SGP-2.

We cloned a mouse homolog of sulfated glycoprotein-2 (SGP-2) cDNA by screening a mouse testicular cDNA library and its nucleotide sequence was determined. The predicted amino acid sequence of the cDNA shares 93% identity with that of rat SGP-2. The nucleotide sequences of both cDNAs show extensive homology throughout the open reading frames and 3' untranslated regions. The 5' untranslated regions, however, share homology only up to 28 bp upstream from the start codons; the rest of sequences are quite different. DNA sequence homology search to mouse SGP-2 cDNA through the EMBL/GenBank database and a recent study on the genomic organization of rat TRPM-2 gene suggest a possibility that there are at least two different SGP-2 mRNAs as a result of alternative splicing and/or different promoter usage in mouse.