腹腔注射2-脱氧-D-葡萄糖可诱发大鼠视上核和室旁核神经元表达Fos

目的 探讨腹腔注射2-脱氧-D-葡萄糖（2-DG）能否激活大鼠下丘脑视上核（SON）和室旁核（PVN）神经元而表达Fos。方法 健康雄性SD大鼠12只,随机分为腹腔注射2-DG组（6只）、生理盐水对照组（3只）及正常对照组（3只）。各自处理后,应用免疫组织化学方法,观察各组下丘脑SON和PVN内Fos表达及其与催产素（OT）和加压素（VP）的双标情况,同时采用ELISA方法对血清中OT和VP的含量进行检测。 结果与生理盐水对照组和正常对照组相比,2-DG引发的特异性Fos免疫阳性产物主要集中分布于下丘脑外侧区和穹隆周区,在SON、PVN也有密集表达。SON和PVN内的Fos表达与该区的特异性神经活性物质OT和VP有共存。OT/Fos双标细胞率（双标细胞占OT阳性细胞的百分率）在SON和PVN分别为87.10%、90.57%,明显高于VP/Fos在这两个核团的双标率（双标细胞占VP阳性细胞的百分率,68.42%、76.92%）,两者比较差异有统计学意义（P<0.05）。ELISA检测结果显示,2-DG组动物血清中OT和VP水平与对照组相比无明显变化。 结论 腹腔注射2-DG可激活大鼠下丘脑SON和PVN内OT和VP神经元表达Fos,SON和PVN可能参与2-DG诱导的急性应激反应。     

The effects of 4-imidazolyl-3-amino-2-butanone (McN-A-1293), a specific histidine decarboxylase inhibitor,on the expression of morphine tolerance and physical dependence in mice

McN-A-1293, a specific histidine decarboxylase inhibitor administered intracisternally in the withdrawal phase, significantly suppressed the expression of morphine tolerance. Severity of withdrawal, assessed by percentage body weight loss, was significantly enhanced although incidence of jumping was unaltered. The opposite effects of this compound on morphine tolerance and withdrawal suggest that tolerance and physical dependence are based on different underlying mechanisms.     

Study of the efferent connections of the enkephalinergic magnocellular dorsal nucleus in the guinea-pig hypothalamus using lesions, retrograde tracing and immunohistochemistry: evidence for a projection to the lateral septum

The efferents of enkephalin-immunoreactive neurons in the magnocellular dorsal nucleus of the guinea-pig were studied using different neuroanatomical methods and indirect immunocytochemical technique. Following unilateral implantation of the fluorescent dye 4',6-diamidino-2-phenylindole in the lateral septal nucleus, retrogradely-labeled perikarya were found in the magnocellular dorsal nucleus. These labeled perikarya reacted with antiserum against enkephalin, demonstrating that enkephalin-immunoreactive neurons in the magnocellular dorsal nucleus project to the lateral septal nucleus. In other experiments, complete bilateral lesions were produced in the magnocellular dorsal nucleus by electrocoagulation. Enkephalin-immunoreactive nerve fibers and terminals were totally depleted in the lateral septal nucleus. This confirms that septal enkephalin-immunoreactive terminals originate in the magnocellular dorsal nucleus and further suggests that this nucleus is the source of all the enkephalin-immunoreactive material found in the septum. Experiments utilizing two different fluorescent dyes, 4',6-diamidino-2-phenylindole and propidium iodide, injected in each side of the lateral septal nucleus, respectively, demonstrated that the magnocellular dorsal nucleus gives off axon collaterals to both sides of the septum, since double-labeling of individual cell bodies was detected in the nucleus. By relating this finding to the results obtained after unilateral destruction of the nucleus, which caused an incomplete loss of enkephalin- immunoreactive material in the lateral septal nucleus ipsilaterally, it is suggested that the enkephalinergic hypothalamo-septal pathway contains unbranching neurons projecting ipsilaterally and branching neurons distributing fibers ipsilaterally and contralaterally. Lesion experiments, and experiments based on the retrograde axonal transport of horseradish peroxidase after intravenous injections, demonstrated that the magnocellular dorsal nucleus contributes neither to the tubero-infundibular nor to the hypothalamo-neurohypophyseal tracts. The lateral septal nucleus receives numerous aminergic and peptidergic projections, indicating the potential importance of this region in physiological and behavioral events. In the guinea-pig, the well-demarcated enkephalinergic pathway demonstrated in this study provides a convenient model for the experimental study of the enkephalinergic innervation of the lateral septal nucleus.     

Regulation of matrix metalloproteinase-2(gelatinase A, MMP-2), membrane-type matrixmetalloproteinase-1 (MT1-MMP) and tissue inhibitorof metalloproteinases-2 (TIMP-2) expression byelastin-derived peptides in human HT-1080 fibrosarcoma cell line

Soluble kappa-elastin peptides were shown to stimulate the expression of MMP-2 (but not MMP-9) byhuman fibrosarcoma HT-1080 cells, both at the protein and mRNA levels; maximal effect being observedat a concentration of 25 mg/ml of kappa-elastin. The stimulatory effect could be reproduced using Val-Gly-Val-Ala-Pro-Gly (VGVAPG) peptide, an elastin-derived hydrophobic hexapeptide which represented theelastin receptor binding sequence of tropoelastin. Furthermore, treatment of cells with lactose (30 mM),which dissociated 67-kDa elastin binding protein (EBP) from cell surfaces, completely abolished this effect,suggesting that the elastin receptor could mediate such a response. Using a specific monoclonal antibody,67-kDa EBP was detected in HT-1080 membrane preparations by Western immunoblotting. Following treat-mentwith 25 mg/ml kappa-elastin or 200 mg/ml VGVAPG, increased levels of the active 62-kDa form ofMMP-2 were found in HT-1080 cell extracts. Stimulation of MT1-MMP mRNA expression by treatmentwith elastin-derived peptides (EDPs) was shown by competitive polymerase chain reaction (PCR). A reversezymography analysis revealed that EDPs also stimulated TIMP-2 (but not TIMP-1) production by HT-1080cells. Competitive PCR confirmed increased TIMP-2 mRNA expression by such treatment. These resultssuggest that occupancy of the 67-kDa elastin receptor by elastin-derived peptides enhanced both expressionand activation of proMMP-2 and consequently, could promote the invasive/metastatic ability of tumor cellsexpressing this receptor. ©Kluwer Academic Publishers     

Biochemical and molecular findings in a patient with myoclonic epilepsy due to a mistarget of the β-glucosidase enzyme

A deficiency of human LIMP-2, a receptor for lysosomal mannose 6-phosphate-independent targeting of the β-glucosidase (βGC), due to mutations in the SCARB2 gene was described only in six families presented with progressive myoclonic epilepsy and nephrotic syndrome. In one of them a mistarget of the βGC was demonstrated.We report here the biochemical and molecular findings in a patient diagnosed with progressive myoclonic epilepsy due to a mistarget of the βGC, probably caused by a LIMP-2 deficiency, providing valuable information for the diagnosis of this rare disorder.     

Dense transient receptor potential cation channel, vanilloid family, type 2 (TRPV2) immunoreactivity defines a subset of motoneurons in the dorsal lateral nucleus of the spinal cord, the nucleus ambiguus and the trigeminal motor nucleus in rat

The transient receptor potential cation channel, vanilloid family, type 2 (TRPV2) is a member of the TRPV family of proteins and is a homologue of the capsaicin/vanilloid receptor (transient receptor potential cation channel, vanilloid family, type 1, TRPV1). Like TRPV1, TRPV2 is expressed in a subset of dorsal root ganglia (DRG) neurons that project to superficial laminae of the spinal cord dorsal horn. Because noxious heat (>52 degrees C) activates TRPV2 in transfected cells this channel has been implicated in the processing of high intensity thermal pain messages in vivo. In contrast to TRPV1, however, which is restricted to small diameter DRG neurons, there is significant TRPV2 immunoreactivity in a variety of CNS regions. The present report focuses on a subset of neurons in the brainstem and spinal cord of the rat including the dorsal lateral nucleus (DLN) of the spinal cord, the nucleus ambiguus, and the motor trigeminal nucleus. Double label immunocytochemistry with markers of motoneurons, combined with retrograde labeling, established that these cells are, in fact, motoneurons. With the exception of their smaller diameter, these cells did not differ from other motoneurons, which are only lightly TRPV2-immunoreactive. As for the majority of DLN neurons, the densely-labeled populations co-express androgen receptor and follow normal DLN ontogeny. The functional significance of the very intense TRPV2 expression in these three distinct spinal cord and brainstem motoneurons groups remains to be determined.     

The cadmium-resistant gene, CAD2, which is a mutated putative copper-transporter gene (PCA1), controls the intracellular cadmium-level in the yeast S. cerevisiae

Yeast cells carrying the CAD2 gene exhibit a resistance to cadmium. We cloned this gene and demonstrated that it was a mutated form derived from the gene of a putative copper-transporting ATPase (PCA1). By site-directed mutagenesis, it appeared that the mutation conferring cadmium resistance was a R970G-substitution in the C-terminal region of Pca1 protein. The intracellular cadmium level of cells carrying CAD2 was lower than that of cells carrying either PCA1 or Δcad2. Furthermore, cells with overexpression of CAD2 showed a much lower intracellular cadmium level than that of cells with a single-copy CAD2. From these results, we conclude that the Cad2 protein controls the intracellular cadmium level through an enhanced cadmium efflux system. Received: 3 September / 9 November 1999     

Association of angiotensin-converting enzyme and angiotensin-converting enzyme-2 gene polymorphisms with essential hypertension in the population of Odisha,India

Background: Hypertension is a serious health issue worldwide and essential hypertension, which includes 90–95% of the cases, is influenced by both genetic and environmental factors. Identification of these factors may help in control of this disease. The Insertion/Deletion (I/D) polymorphism in Angiotensin-Converting Enzyme (ACE) gene and rs2106809 (C?>?T) polymorphism in Angiotensin-Converting Enzyme 2 (ACE2) gene have been reported to be associated with essential hypertension in different populations.

Aim: To investigate the association of ACE I/D and ACE2 rs2106809 polymorphisms with essential hypertension in the population of Odisha, an eastern Indian state.

Subjects and methods: A total of 246 hypertensives (159 males and 87 females) and 274 normotensives (158 males and 116 females) were enrolled in the study. Detailed anthropometric data, tobacco, alcohol and food habits were recorded and 2?ml of venous blood was collected for biochemical and genetic analysis.

Results: The DD genotype of ACE and TT genotype of ACE2 were significantly high among female hypertensives, while T allele of ACE2 was linked to male hypertensives. In the male population, alcohol was also identified as a potential risk factor.

Conclusion: Among females, ACE I/D and ACE2 rs2106809 polymorphisms, while among males, ACE2 rs2106809 polymorphism and alcohol consumption are associated with essential hypertension in the study population.     

Interstitial deletion of the long arm of chromosome 2 in a malformed infant with karyotype 46, XX,del(2) (q31q33)

We describe a malformed newborn girl with an interstitial deletion of the long arm of chromosome 2 (karyotype: 46, XX, del (2) (q31q33)). This is the first report of this particular chromosome abnormality that includes autopsy findings. Comparison with previous cases in the literature suggests that this particular deletion uniformly results in developmental delays, craniofacial changes, and occasionally results in microcephaly, low-set ears, and hand and foot abnormalities.     

2型糖尿病思看血清脂联素、胰岛素、G-反应蛋白、瘦素水平变化分析

目的:通过对2型糖尿病患者血清中脂联素(APN)、胰岛素、C-反应蛋白、瘦素(Leptin)、胰岛素抗体及谷氨酸脱羧酶抗体(GAD-Ab)检测分析,探讨胰岛β细胞功能和胰岛素抵抗的机理,为其早期诊断治疗提供依据。方法:用化学发光、酶联免疫分析、放射免疫分析对184例2型糖尿病患者和30例正常对照者及75例DM2治疗观察组经一年治疗前后水平检测比较分析。结果:正常对照组与DM2比较胰岛素、瘦素、C-反应蛋白、胰岛素抗体有显著性差异(P<0 01),且呈正相关,与脂联素比较有显著性差异(P<0 001)但呈负相关;75例治疗后除脂联素、胰岛素抗体外其它几项指标较治疗前明显下降,有统计学意义,而脂联素较治疗前明显升高(P<0 001)有显著性差异。结论: 2型糖尿病标志物的检测,对其早期诊断、治疗预后及生理机制的研究具有重要意义。     

Biallelic variants in ribonuclease inhibitor (RNH1), an inflammasome modulator,are associated with a distinctive subtype of acute,necrotizing encephalopathy

PurposeMendelian etiologies for acute encephalopathies in previously healthy children are poorly understood, with the exception of RAN binding protein 2 (RANBP2)–associated acute necrotizing encephalopathy subtype 1 (ANE1). We provide clinical, genetic, and neuroradiological evidence that biallelic variants in ribonuclease inhibitor (RNH1) confer susceptibility to a distinctive ANE subtype.MethodsThis study aimed to evaluate clinical data, neuroradiological studies, genomic sequencing, and protein immunoblotting results in 8 children from 4 families who experienced acute febrile encephalopathy.ResultsAll 8 healthy children became acutely encephalopathic during a viral/febrile illness and received a variety of immune modulation treatments. Long-term outcomes varied from death to severe neurologic deficits to normal outcomes. The neuroradiological findings overlapped with ANE but had distinguishing features. All affected children had biallelic predicted damaging variants in RNH1: a subset that was studied had undetectable RNH1 protein. Incomplete penetrance of the RNH1 variants was evident in 1 family.ConclusionBiallelic variants in RNH1 confer susceptibility to a subtype of ANE (ANE2) in previously healthy children. Intensive immunological treatments may alter outcomes. Genomic sequencing in children with unexplained acute febrile encephalopathy can detect underlying genetic etiologies, such as RNH1, and improve outcomes in the probands and at-risk siblings.     

Molecular analysis of the 5α-steroid reductase type 2 gene in a family with deficiency of the enzyme

This report describes the identification of a point mutation in the 5α-reductase type 2 (5α-SR2) gene from a family in which both sibs (6 and 3 years old) have steroid 5α-reductase 2 deficiency. The five exons of the gene were individually amplified by the polymerase chain reaction (PCR) and analysed for single-strand conformation polymorphisms (SSCP) to detect mutations. Direct sequencing of the mutant PCR products demonstrated a single C→T mutation, within exon 4, changing codon 227 from CGA (Arg) to TGA (premature termination signal). Both patients were homozygous for the mutation, but their parents were heterozygous. These results suggest that the mutation at codon 227 impairs normal 5α-SR2 function, thus leading to the phenotypical expression of this rare enzymatic defect. Am. J. Med. Genet. 69:69–72, 1997. © 1997 Wiley-Liss, Inc.     

New data on the morphology of Nilonema senticosum (Nematoda, Philometridae), a parasite of Arapaima gigas (Pisces), with notes on another philometrid, Alinema amazonicum, in Peru

Female specimens of a little-known philometrid nematode, Nilonema senticosum (Baylis, 1927), were collected from the swimbladder of the arapaima, Arapaima gigas, from the Amazon River basin (Iquitos, Loreto District) in Peru. Scanning electron microscopical (SEM) examination, used for the first time in this species, made it possible to reveal some taxonomically important, previously unreported features of N. senticosum, such as the presence of minute cephalic papillae (10 papillae in 2 circles) and amphids surrounding the small oral aperture, and to confirm the absence of an anal opening in this species. Males and females of another philometrid, Alinema amazonicum (Travassos, 1960), were recovered from the body cavity of the pimelodid catfishes Calophysus macropterus and Brachyplatystoma juruense (a new host record) from the Amazon River basin (fish market in Iquitos, Loreto District) in Peru. SEM examination, not previously used in the male of A. amazonicum, enabled to study in detail the male cephalic and caudal structures. An erratum to this article is available at .     

儿童肺结核病患者血清IL-2、SIL-2R和VEGF水平及临床意义

目的:探讨了儿童肺结核病患者血清白细胞介素-2(IL-2)、可溶性白细胞介素-2受体(SIL-2R)和血管内皮生长因子(VEGF)水平的变化及意义。方法:分别应用放射免疫分析和ELISA对68例儿童肺结核病患者进行了血清IL-2、SIL-2R和VEGF水平观察,并与35名正常健康人作比较。结果:活动性儿童肺结核病患者血清IL-2水平显著低于正常儿组(P<0.01),而SIL-2R和VEGF水平显著高于正常儿组(P<0.01)。结论:测定儿童肺结核病患者血清IL-2、SIL-2R和VEGF水平更有助于儿童肺结核病活动性的检测,对该病的预防和预后均有一定的意义。     

A novel EXT2 mutation in a consanguineous family with severe developmental delay,microcephaly, seizures,feeding difficulties,and osteopenia extends the phenotypic spectrum of autosomal recessive EXT2-related syndrome (AREXT2)

We report a consanguineous family where 2 boys presented with developmental delay, hypotonia, microcephaly, seizures, gastro-intestinal abnormalities, osteopenia, and neurological regression. Whole exome sequencing performed in one of the boys revealed the presence of a novel homozygous missense variant in the EXT2 gene: c.11C?>?T (p.Ser4Leu). Segregation analysis by Sanger sequencing confirmed homozygous by descent autosomal recessive transmission of this mutation. Another family was previously reported with homozygous mutations in this gene in four siblings affected with a nearly similar clinical condition (Farhan et al., 2015). We discuss the similarities and differences between the two syndromes and propose AREXT2 as a new acronym for EXT2-related diseases.     

Genetic etiology of Parkinson disease associated with mutations in the SNCA,PARK2, PINK1, PARK7, and LRRK2 genes: a mutation update

To date, molecular genetic analyses have identified over 500 distinct DNA variants in five disease genes associated with familial Parkinson disease; α‐synuclein (SNCA), parkin (PARK2), PTEN‐induced putative kinase 1 (PINK1), DJ‐1 (PARK7), and Leucine‐rich repeat kinase 2 (LRRK2). These genetic variants include ～82% simple mutations and ～18% copy number variations. Some mutation subtypes are likely underestimated because only few studies reported extensive mutation analyses of all five genes, by both exonic sequencing and dosage analyses. Here we present an update of all mutations published to date in the literature, systematically organized in a novel mutation database ( http://www.molgen.ua.ac.be/PDmutDB ). In addition, we address the biological relevance of putative pathogenic mutations. This review emphasizes the need for comprehensive genetic screening of Parkinson patients followed by an insightful study of the functional relevance of observed genetic variants. Moreover, while capturing existing data from the literature it became apparent that several of the five Parkinson genes were also contributing to the genetic etiology of other Lewy Body Diseases and Parkinson‐plus syndromes, indicating that mutation screening is recommendable in these patient groups. Hum Mutat 31:763–780, 2010. © 2010 Wiley‐Liss, Inc.