[Expression of Glutathione Peroxidases and Its Effect on Clinical Prognosis in Glioma Patients].

Objective To investigate the relationship between the expression of glutathione peroxidase(GPX)genes and the clinical prognosis in glioma patients,and to construct and evaluate the model for predicting the prognosis of glioma. Methods The clinical information and GPX expression of 663 patients,including 153 patients of glioblastoma(GBM)and 510 patients of low-grade glioma(LGG),were obtained from The Cancer Genome Atlas(TCGA)database.The relationship between GPX expression and patient survival was analyzed.The key GPX affecting the prognosis of glioma was screened out by single- and multi-factor Cox's proportional-hazards regression models and validated by least absolute shrinkage and selection operator(Lasso)regression.Finally,we constructed the model for predicting the prognosis of glioma with the screening results and then used concordance index and calibration curve respectively to evaluate the discrimination and calibration of model. Results Compared with those in the control group,the expression levels of GPX1,GPX3,GPX4,GPX7,and GPX8 were up-regulated in glioma patients(all P<0.001).Moreover,the expression levels of other GPX except GPX3 were higher in GBM patients than in LGG patients(all P<0.001).The Kaplan-Meier curves showed that the progression-free survival of GBM with high expression of GPX1(P=0.013)and GPX4(P=0.040),as well as the overall survival,disease-specific survival,and progression-free survival of LGG with high expression of GPX1,GPX7,and GPX8,was shortened(all P<0.001).GPX7 and GPX8 were screened out as the key factors affecting the prognosis of LGG.The results were further used to construct a nomogram model,which suggested GPX7 was the most important variable.The concordance index of the model was 0.843(95%CI=0.809-0.853),and the calibration curve showed that the predicted and actual results had good consistency. Conclusion GPX7 is an independent risk factor affecting the prognosis of LGG,and the nomogram model constructed with it can be used to predict the survival rate of LGG.

Aminophylline restores glucocorticoid sensitivity in a guinea pig model of sudden sensorineural hearing loss induced by lipopolysaccharide.

Glucocorticoids have been used to treat hearing loss and vestibular dysfunction for many years. However, some reports have indicated that a subset of patients with these disorders exhibit glucocorticoid insensitivity or resistance. A reduction in histone deacetylase 2 (HDAC2) activity and expression has been reported to play a critical role in glucocorticoid resistance. Here, we investigated the protective effects of aminophylline on HDAC2 expression and glucocorticoid sensitivity in lipopolysaccharide (LPS)-induced sudden sensorineural hearing loss in guinea pigs. We assessed hearing recovery in LPS-applied guinea pigs, which were either left untreated or were systemically treated with either dexamethasone, aminophylline, or a combination of the two. We utilized fluorescence microscopy and enzyme-linked immunosorbent assay to analyze the distribution patterns of HDAC2 and detect its levels in the cochlea. We used hematoxylin-eosin staining to examine cochlear histopathological changes. In the absence of treatment, significant hearing loss was detected in LPS-exposed animals. A synergistic effect was observed between aminophylline and dexamethasone in maintaining HDAC2 expression levels, preventing hearing loss in LPS-exposed animals and reducing cochlear damage. This study indicates that aminophylline can restore glucocorticoid sensitivity, which provides a new approach to treating patients with hearing disorders who are refractory to glucocorticoids.

TIMP-1 polymorphisms in a Chinese Han population with intracerebral hemorrhage.

Remodeling of extracellular matrix (ECM) and breakdown of blood-brain barrier (BBB) are crucial events in the pathogenesis of intracerebral hemorrhage (ICH). Matrix metalloproteinases (MMPs), particularly MMP-9 and MMP-2, are the most important degrading enzymes in the ECM and BBB. These proteolytic effects are controlled predominantly by tissue inhibitors of metalloproteinases (TIMPs). TIMP-1 is the main endogenous inhibitor of MMP-9. Two polymorphisms in the TIMP-1 gene (rs4898 and rs2070584) were selected through a literature review and successfully genotyped in a study sample of 410 ICH patients and 305 controls. Differences in genotype and allele frequencies of identified polymorphisms were determined. Furthermore, the serum levels of TIMP-1 were measured in a subgroup of 96 ICH patients on days 1 after ICH onset and 76 controls. Analyses showed that C allele of rs2070584 was significantly associated with the development of ICH in male subjects (p = 0.037, OR = 1.535, 95%CI 1.025-2.300). Multiple logistic regression analysis under three genetic models demonstrated both rs4898 and rs2070584 were not risk factors for ICH in female subjects. Furthermore, serum levels of TIMP-1 were significantly higher in ICH patients than those in normal controls. However, the serum levels of TIMP-1 showed a nonsignificant decrease, depending on the alleles and genotypes of rs2070584 both in male and female cases. In conclusion, this is the first association study of the TIMP-1 gene variants with ICH. Our data suggest that C allele of rs2070584 is a risk factor for ICH development in the Chinese male population. However, the precise function of this variant needs further investigation.

Neuroprotection by silencing iNOS expression in a 6-OHDA model of Parkinson's disease.

In this study, we investigated the protective role of silenced iNOS expression in neuron death in the nigrostriatal pathway in a 6-OHDA animal model of Parkinson's disease (PD). The animal model was established by intrastriatal infusion of a single dose of 6-OHDA. Silencing of iNOS expression was established by intrastriatal infusion of adenovirus-carried iNOS-targeted small interfering RNA (siRNA). Apomorphine-induced rotation behavior was measured. Expression of iNOS, OX-42, and TH; levels of DA, DOPAC, and HVA in the striatum; and the levels of p53 phosphorylation, Bax, and cleaved caspase-3 (CC3) in the substantia nigra were measured. We demonstrated that co-infusion of 6-OHDA with adenovirus expressing siRNA of iNOS blocked the activation of microglia, iNOS transcription, and p53-Bax-CC3 apoptotic cascade as well as significantly blocking 6-OHDA-induced decreases in DA, DOPAC, HVA, and TH levels and effectively decreased rotation number. Our study highlighted the role of iNOS in the neurodegeneration of nigrostriatal dopaminergic neurons in the 6-OHDA animal model of PD and suggested that the microglial activation-iNOS-p53-Bax-CC3 apoptotic pathway plays a key role in the neurodegeneration in the 6-OHDA model.

Infusion of BDNF into the nucleus accumbens of aged rats improves cognition and structural synaptic plasticity through PI3K-ILK-Akt signaling.

To investigate the involvement of the nucleus accumbens (NAc) in cognitive impairment and the therapeutic effects of brain-derived neurotrophic factor (BDNF) in an animal model of cognitive deficit, we infused BDNF into the NAc of cognitively impaired aged rats. Cognition was evaluated by Morris water maze test. Structural synaptic plasticity was measured by Golgi staining. Brain tissue homogenization was used to measure the changes in signal molecules. Cultured PC-12 cells expressing tyrosine kinase receptor (Trk) B/p75 neurotrophin receptor (p75(NTR)), p75(NTR) or TrkA/p75(NTR) receptors were used for BDNF stimulation assays. Significant decreases in the levels of BDNF, phosphatidylinositol-3-kinase (PI3K) and integrin-linked kinase (ILK) activity, protein kinase B (Akt) Ser47³ phosphorylation, dendritic branching, and density of dendritic spines on medium spiny neurons were observed in the NAc. Importantly, infusion of BDNF restored cognition, synaptic plasticity, and cell signaling. In cultured PC-12 cells, BDNF activated PI3K/Akt signaling through the TrkB receptor, whereas stimulation of ILK/Akt occurred through TrkA/p75(NTR) heteroreceptor. Our study suggested that the decreased BDNF level and its downstream signaling as well as loss of synaptic plasticity in the NAc are associated with cognitive impairments in aged rats. The BDNF-activated PI3K-Akt and ILK-Akt signaling play a key role in structural synaptic plasticity. Our study also suggested that BDNF could be a mechanism-based treatment for dementia.

[Association of single nucleotide polymorphisms of kallikrein 1 gene with cerebral hemorrhage in Changsha Han Chinese].

OBJECTIVE: To explore the association between single nucleotide polymorphisms (SNPs) of KLK1 gene and cerebral hemorrhage in Changsha Han Chinese. METHODS: Two hundred and seventy-three cerebral hemorrhage (CH) patients and 140 healthy controls were collected. The SNPs of rs5516 and rs5517 loci of KLK1 gene were analyzed by SNaPshot methods and direct sequencing. RESULTS: (1)Genotype and allele frequencies in rs5516 locus had no difference between the CH patients and controls (P> 0.05). However, the A allele frequency of the rs5517 locus in CH patients was higher than that in the control group (0.419, 0.321 respectively, P< 0.05). (2)In the control group,the levels of diastolic blood pressure (DBP) of the GA and AA genotype carriers of the rs5517 locus were significantly higher than those of the GG genotype (P< 0.05), while the levels of blood pressure were not significantly different among different genotypes of the rs5516 polymorphism in both CH patients and the control group(P> 0.05). CONCLUSION: Author's preliminary results suggested that the rs5517 polymorphism was associated with cerebral hemorrhage, while the rs5516 polymorphism was not in Changsha Han Chinese.

Association between Val/Leu(247) polymorphism of apolipoprotein H and cerebral infarction in a Chinese population.

BACKGROUND: Antiphospholipid antibodies (aPL) are considered to be a cause of an acquired hypercoagulable state leading to cerebral infarction (CI). Apolipoprtein H (apoH) is an important target antigen for aPL and thus apoH polymorphisms may influence aPL production and the development of CI. The purpose of this study was to identify associations between the Val/Leu(247) polymorphism of apoH gene and CI in a Chinese cohort. METHODS: This study comprised 130 CI patients and 100 healthy control subjects. Polymorphism assignment was determined by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique and DNA sequencing. The presence of aPL was detected by ELISA utilizing irradiated ELISA plates. RESULTS: Our results demonstrated an association between the Val/Leu(247) polymorphism of apoH gene and aPL in CI patients. The frequency of V allele was significantly higher in aPL-positive CI patients compared with control group (chi(2) = 6.864, P < 0.05). VL genotype frequency was also significantly higher in aPL-positive CI group compared with control group (chi(2) = 13.879, P < 0.05) and aPL-negative CI group (chi(2) = 5.567, P < 0.05). CONCLUSIONS: The Val(247) allele of apoH gene is significantly associated with the presence of aPL in Chinese patients with CI.

[The association of apolipoprotein B gene polymorphism with cerebral infarction with positive family history and its effect on plasma lipid levels].

OBJECTIVE: To explore the relationship between apolipoprotein B (apoB) gene G12669A polymorphism and cerebral infarction with family history, and to evaluate the effect of G12669A polymorphism on plasma lipid levels. METHODS: Peripheral blood samples were collected from 147 members of 15 cerebral infarction families, including 47 cerebral infarction patients with positive family history (CIFH-P), 43 first-degree relatives (CIFH-I), 28 second-degree relatives (CIFH-II), and 29 third-degree relatives (CIFH-III), 83 sporadic cerebral infarction (SCI) patients, and 100 healthy controls. Polymerase chain reaction- restriction fragment length polymorphism was used to detect the apoB gene G12669A polymorphism. Oxidase method was used to detect the levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL). The serum levels of lipoprotein (a) [LP (a)], apoB-100, and apoAI were determined by immune method. RESULTS: (1) The frequencies of A allele in the CIFH-P, CIFH-I, CIFH-II, CIFH-III, and SCI groups patients and control group were 0.106, 0.081, 0.036, 0.034, 0.090, and 0.045 respectively, that of the CIFH-P group being significantly higher than that of the control group (P < 0.05), and those of the CIFH-I, CIFH-II, CIFH-III, and SCI groups not being significantly different from that of the control group. (2) In both CIFH-P and SCI groups, the TC and LDL-C levels of the patients with G/A gene type were significantly higher than those of the G/G gene type, while the HDL-C level of the patients with G/A gene type was significantly lower than that of the G/G gene type (all P < 0.05). CONCLUSION: A allele in G12669A polymorphism may be one of the genetic factors influencing the susceptibility to CI in the individuals with a positive family history, and it may play its role through its influence on the blood lipid levels.

[Effects of cyclin dependent protein kinase inhibitor olomoucine on the neuronal apoptosis after status epilepticus: experiment with rats].

OBJECTIVE: To investigate the effects of olomoucine, a cyclin dependent protein kinase (CDK) inhibitor, on the neuronal apoptosis after status epilepticus (SE). METHODS: Lithium chloride was injected intraperitoneally, and pilocarpine was injected intraperitoneally after 18 h to 24 SD rats so as to cause SE. Twenty-two of the 24 rats developed SE and 2 of them died. The surviving 20 rats were then randomly divided into 2 equal groups: olomoucine group, injected intracerebroventricularly after the SE was terminated by diazepam and chloral hydrate once a day for 3 days, and SE group, infused intracerebroventricularly with DMSO solution Another 10 rats were injected intraperitoneally with normal saline and then infused intracerebroventricularly with DMSO solution to be used as control group. Six hours after SE attack 5 rats from each group were killed respectively with their brains taken out. Semiquantitative RT-PCR was used to detect the mRNA expression of anti-inflammatory cytokines, such as interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha. Three days later the other 5 rats in each group were killed with their entorhinal cortex and hippocampus taken out. TUNEL was used to observe the apoptosis. Immunofluorescence (IF) staining was used to detect the expression of neuronal nuclear nucleoprotein (NeuN) and cyclin B1. RESULTS: TUNEL showed that apoptotic neurons were rare in the control group and were numerous in the SE group, especially in the entorhinal cortex and the hylus of dentate gyrus, and the number of apoptotic neurons in the hylus of dentate gyrus of the olomoucine group was not significantly different from that of the control group (P < 0.05), however, the number of apoptotic cells in the entorhinal cortex of the olomoucine group was still significantly higher than that of the control group (P < 0.05). IF staining demonstrated that in the control group the co-expression of NeuN and TUNEL-labeled cells was weak; and in the SE group the co-expression of NeuN and TUNEL was significantly increased compared with that in the control group (P < 0. 05). The number of cyclin B1 positive cells in the olomoucine group was 18.22 +/- 3.99, significantly lower than that of the SE group (24.57 +/- 6.78, P < 0.05). Semiquantitative RT-PCR showed that the IL-1beta and TNF-alpha mRNA expression levels of the SE group were both significantly higher than those of the control group (both P < 0.05), and the IL-1beta and TNF-alpha mRNA expression levels of the olomoucine group, except the TNF-alpha mRNA expression in the cortex, were all significantly lower than those of the SE group (all P < 0.05), and not significantly different from those of the control group (all P > 0.05). CONCLUSION: Olomoucine treatment can inhibits cell cycle protein B1 expression, anti-inflammatory cytokines such as IL-1beta and TNF-alpha secretion, thus decreasing neuronal death and providing neuroprotection after SE, which suggests a potential promising therapeutic way for epilepsy treatment.

[Isolation of Malassezia furfur from the groin abscess of a renal transplant patient].

We report here the discovery of Malassezia furfur from a groin abscess of a renal transplant patient. A 33-year-old male patient was admitted to our hospital because of a high fever and a persistent inflammatory nodule on his right groin for one week. He had received a renal transplant 3 years before and remained on immunosuppressive agents. He was treated with broad-spectrum antibiotics after hospitalization but the nodule formed a large abscess and then a deep ulcer instead of resolving. Examination of the culture by light microscopy revealed ovoid budding yeasts displaying collar-shaped structure. Subculture of the primary colonies onto Sabouraud's dextrose agar and medium containing rapeseed oil resulted in growth only on the medium containing rapeseed oil. All of the isolates was identified as Malassezia furfur. The pathogenicity of the isolates was tested in mice by intravenous injection of (3-5) x 10(8) cfu per mouse after immunosuppression with 500 mg/kg of prednisone intraporitoneally on day-2. In the mouse model, micro-abscess and inflammatory reaction and oval yeasts with budding were noted in histopathologic section of the viscera of the mice. A rib-like or serrate-like structure of the inner side of cell wall, characteristic for Malassezia spp., was observed by transmission electron microscopy. The patient received oral fluconazole and topical amphotericin B. The isolate before antifungal therapy was sensitive to both fluconazole and amphotericin B, while the isolate after antifungal treatment was only sensitive to amphotericin B. Proteinase activity of the isolates increased 1.43 times after antifungal treatment. This case indicated the invasive power of M. furfur in deep infection. Renal transplantation and reception of long-term immunosuppressive treatment are risk factors for the invasive infection of this fungus.

[Protective effect of Polygonum multiflorum thunb on the cerebral cholinergic neurofibers in rats].

OBJECTIVE: To observe the number and morphological changes of cholinergic neurofibers in rats under the destruction of kainic acid, and to investigate the protective effect and the mechanism of polygonum multiflorum thunb (PMT) on acetyl-chorine esterase (AChE) neurofibers. METHODS: Excitative neurotoxin kainic acid was injected into the basal forebrain, Meynert neucleus, medial septal neucleus and the Broca neucleus to establish the destruction model. Then the destructive experimental group was fed with PMT, and the histochemical method was used to display the changes of AChE fibers, the protection and activation of PMT. RESULTS: The number of projecting AChE fibers to the cortex and the hippocampus in PMT group was larger than that in the control group, and no morphological destruction was seen in the experimental group (P < 0.01). CONCLUSION: PMT has a protective effect on AchE projecting fibers in rats.