A cache coherence protocol is a vital component of a multiprocessor to maintain the data consistency. In this paper, we proposed VFCC, which is a simulation framework to validate a cache-coherence protocol implementation of a commercial 64-bit superscalar multiprocessor. It exploits multiple-level parallelism to accelerate validation without overheads among threads. Our experimental results demonstrate VFCC has a 5.0× speedup than a traditional simulator on a conventional 16-core host machine.

Analysis of global methylation in cells has revealed correlations between overall DNA methylation status and some biological states. Recent studies suggest that epigenetic regulation through DNA methylation could be responsible for neuroadaptations induced by addictive drugs. However, there is no investigation to determine global DNA methylation status following repeated exposure to addictive drugs. Using mice conditioned place preference (CPP) procedure, we measured global DNA methylation level in the nucleus accumbens (NAc) and the prefrontal cortex (PFC) associated with drug rewarding effects. We found that cocaine-, but not morphine- or food-CPP training decreased global DNA methylation in the PFC. Chronic treatment with methionine, a methyl donor, for 25 consecutive days prior to and during CPP training inhibited the establishment of cocaine, but not morphine or food CPP. We also found that both mRNA and protein level of DNMT (DNA methytransferase) 3b in the PFC were downregulated following the establishment of cocaine CPP, and the downregulation could be reversed by repeated administration of methionine. Our study indicates a crucial role of global PFC DNA hypomethylation in the rewarding effects of cocaine. Reversal of global DNA hypomethylation could significantly attenuate the rewarding effects induced by cocaine. Our results suggest that methionine may have become a potential therapeutic target to treat cocaine addiction.

It has been shown that gamma -irradiation induces apoptosis of the human promyeloid leukemia cell line HL-60, but the mechanism remains unclear. To explore the effect of caspase-3 in this apoptotic model, antisense oligodeoxynucleotides (ASODNs) targeting 5'-noncoding region (ASODN-1) and initial translation region (ASODN-2) of caspase-3 mRNA were designed, synthesized and introduced into HL-60 cells by means of liposome-mediated transfection followed by gamma-irradiation in the present study. The TUNEL assay was used for morphological analysis of HL-60 cell apoptosis. Immunocytochemical staining, Western blotting and RT-PCR were, respectively, performed for detecting expression of caspase-3 and its mRNA. HL-60 cells transfected with mismatched oligodeoxynucleotide (MODN) or untransfected were taken as the control groups. The TUNEL assay showed that the percentages of HL-60 cell apoptosis induced by gamma-Irradiation in both ASODN-1 and ASODN-2 groups were significantly reduced compared with those in the control group (P<0.01) when the final transfection concentration was > or = 3 micromol/L. Immunocytochemistry demonstrated that in the ASODN-1 and ASODN-2 groups, caspase-3 positive cell percentages were reduced and average gray values of the positive cells increased significantly compared with those in the control group (P<0.01). Western blotting found that procaspase-3 expression in HL-60 cells of the ASODNs groups was decreased,and it was lower in the ASODN-1 group than in the ASODN-2 group. RT-PCR revealed marked expression of caspase-3 mRNA in HL-60 cells of the control group. Expression of caspase-3 mRNA was decreased after ASODNs transfection. Furthermore, ASODN-1 was more effective in inhibiting HL-60 cell apoptosis (P<0.05) and caspase-3 expression (P<0.01) than ASODN-2. These results indicate that caspase-3 mRNA ASODNs prevent HL-60 cells from apoptosis induced by gamma-radiation,and reduce expression of caspase-3 and its mRNA. These effects are dose dependent in a certain range.

Objective To study the inhibitory effects of caspase-3 mRNA antisense oligodeoxynucleotides (ASODN) on expressions of caspase-3 and it's mRNA in γ-radiation induced apoptotic HL-60 cells, and screen the effective ASODN. Methods ASODN-1 and ASODN-2 targeting 5'-noncoding region and initial translation region of caspase-3 mRNA were respectively designed, synthesized and introduced into HL-60 cells by means of liposome-mediated transfection followed by 10 Gy γ-radiation exposures. TUNEL assay was conducted to investigate the morphologic change and apoptotic percentage of HL-60 cells 18 h later. Immunocytochemical staining and one step RT-PCR were respectively performed to detect the expressions of caspase-3 and it's mRNA. Mismatched oligodeoxynucleotide (MODN) transfected and un-transfected HL-60 cells were taken as control. Results TUNEL assay found that the apoptotic percentages in ASODN- 1 and ASODN-2 groups were significantly reduced compared with the control groups (P＜0.01) when the final concentration of both ASODNs was ≥3 μmol/L. Immunocytochemistry showed that caspase-3 positive cell percentages were reduced but the average gray values increased significantly compared with the control groups (P＜0.01). RT-PCR showed expressions of caspase-3 mRNA was decreased after ASODN transfection. Furthermore, ASODN-1 proved more effective in inhibiting HL-60 cell apoptosis than ASODN-2 (P＜0.01). Conclusion Caspase-3 mRNA ASODNs can prevent HL-60 cells from apoptosis induced by γ-radiation and reduce expression of caspase-3 and its mRNA. These effects are dose dependent in a certain range.

To detect the changes in glial cells in the cerebellum of Niemann-Pick disease type C (NPC). The antibodies to cathepsin D (Cat-D) and glial fibrillary acid protein (GFAP) and immunohistochemical PAP technique were used to observe the changes in microglia and astrocytes in the cerebella from NPC and normal mice of different ages. The results showed that the densely Cat-D-stained microglia were progressively increased in number and became larger with age in the cerebellum,especially in the cortical molecular layer of NPC mice. Similarly, the GFAP-positive astrocytes in the cerebellar medulla and granular layer were also swollen and increased in number. Bergmann cells in the cerebellar cortex gradually increased and their radial processes reached the surface of the molecular layer. Only mild gliosis was found in the nodulus and uvula vermis. The results indicated that microglia and astrocytes in the cerebellum of the NPC mouse were markedly activated. They may correlate with neurodegeneration of Purkinje cells and other types of neurons.

The study describes that 0.5% Adriamycin was injected into the infraorbital branch of trigeminal nerve in rats and the drug was swiftly transported by retrograde axoplasmic flow into the sensory neuron's counterpart. As a result, Adriamycin specific autofluorescence was seen in trigeminal ganglion. Degeneration and necrosis of the cells were shown under light and electronic microscope, and the trigeminal brainstem evoked potentials became unobtainable. The authors believe that the technique is an uncomplicated, safe and reliable method for dealing with trigeminal neuralgia.

A well-defined, gonadotropin-releasing hormone (GnRH)-containing fiber pathway, the ventral hypothalamic tract (VHT), is described by immunostaining in fetal rhesus macaques (109-156 days gestation). The VHT arises above the lateral aspects of the optic chiasm near the supraoptic nucleus, and courses ventromedially close to the ventral hypothalamic surface to terminate in the infundibulum and zona externa of the median eminence. It is formed by the confluence of GnRH-immunopositive (GnRH+) axons from local neurons, from a few GnRH+ cells in the inferior thalamic peduncle, and probably from more anterior neurons in the septum and preoptic area. Bipolar GnRH+ neurons contributing directly to the VHT are grouped at its origin dorsolateral to the optic chiasm, dorsal and medial to the optic tracts, at the infundibular lip, and within the pathway between. At the infundibular lip, GnRH+ perikarya are generally lateral or ventral to the infundibular (arcuate) nucleus, and are rarely within the nucleus itself. Cell bodies here are sometimes tripolar, but GnRH+ intercellular contacts are seldom seen. A few VHT fibers extend to the ventral surface of the brain just beneath the pia mater. Abundant capillaries in the subarachnoid space suggest a possible route for delivery of GnRH to the adenohypophysis in early gestation, before maturation of the hypophysial portal system occurs. Posterior to the infundibulum, a few VHT fibers are joined by descending periventricular fibers forming a dense fiber band beneath the premammillary recess of the third ventricle. Totals of GnRH+ cell bodies in the prosencephalon of the fetal rhesus macaque are estimated to be 5,600 in females (n = 2) and 2,600 in males (n = 3). More than 60% of VHT neurons are located in the medial basal hypothalamus, and the majority of basal hypothalamic GnRH+ neurons (86%) are associated with the VHT. Furthermore, reports of the autonomy of the medial basal hypothalamic-hypophysial unit in control of gonadotropin secretion suggest that the VHT may be the most important GnRH system involved in primate reproduction. It is clear that fetal material may offer the best model to study the GnRH neuronal system in primates.

The structure of the prohormone for mammalian gonadotropin releasing hormone (proGnRH) includes the GnRH decapeptide followed by a 56 amino acid GnRH-associated peptide (GAP). In this study, we compared immunostaining of brain neurons and fibers for GAP and GnRH in fetal rhesus monkeys and juvenile baboons. We used antisera against different portions of human and rat GAP (proGnRH 14-24, proGnRH 40-53, and proGnRH 52-66) or against GnRH and the PAP technique. Liquid phase absorption with GAP or GnRH confirmed the specificity of these antisera. Major accumulations of GAP immunoreactive (GAP+) perikarya occurred in the medial septal and preoptic areas and the nucleus of the diagonal band of Broca (44.6% in rhesus, 49.6% in baboon), supraoptic region including the area dorsal to the optic tract (21.9% in rhesus, 23.0% in baboon), and the medial basal hypothalamus (15.7% in rhesus, 16.4% in baboon), especially at the infundibular lip. Occasional cell bodies were scattered throughout the hypothalamic and forebrain regions studied. GAP+ fibers were widely distributed, but formed well-defined pathways such as the periventricular and ventral hypothalamic tract. In addition, GAP+ nerve terminals with various densities occurred in the lamina terminalis, the zona externa of the infundibulum, and behind the infundibular stalk. Fetal rhesus macaques had more GAP+ cell bodies, denser fiber networks, and more distinct pathways than juvenile baboons. However, fiber and terminal immunostaining was somewhat less intense for GAP than GnRH in comparable regions. These results indicate that proGnRH (GAP) is present in the same population of neurons as GnRH in the primate brain. They also suggest that post-translational products of proGnRH are present in perikarya, axons and terminals, and that GnRH and GAP and/or further cleavage products are consecreted into hypophysial portal blood in the primate.

The distribution of serotonin (5-HT) and tyrosine hydroxylase (TH) was examined in the hypothalamus of juvenile baboons, 24 h after infundibular stalk section. Simultaneous immunostaining for 5-HT with peroxidase-antiperoxidase (PAP) and TH with 15 nm colloidal gold (IGS) was performed on Vibratome sections from 3 operated and 1 control female. Light microscopy revealed fine 5-HT immunopositive (5-HT+) fibers, presumably axons, in the suprachiasmatic nuclei and ventromedial hypothalamus (VMH) after stalk section. In addition, focal accumulations of swollen and heavily stained 5-HT+ fibers occurred on the side of the surgical approach. Enlarged fibers were densest in the medial preoptic area, lateral and VMH areas, and the median eminence. TH immunoreactivity (TH+) in VMH cell bodies and axons was only slightly increased over that in controls. Electron microscopy of areas of 5-HT+ and TH+ overlap (medial VMH and adjacent periventricular zone) showed that 5-HT+ profiles were mostly unmyelinated axons and irregular varicosities. A few myelinated 5-HT+ axons were also observed. TH+ perikarya, dendrites, axons and terminals showed gold labeling characteristic for this enzyme. However, colocalization of 5-HT (PAP) and TH (IGS) was present in a number of fiber varicosities in experimental animals only. Both single- and double-labeled profiles occurred in individual thin sections, thus arguing against antibody cross-reactivity. These results indicate that: hypothalamic 5-HT+ fibers project to the median eminence in primates; 5-HT fibers become more obvious after stalk section due to accumulation of transmitter; focal 5-HT+ immunoreactivity in the hypothalamus can increase dramatically after distant and mild surgical trauma, and coexistence of 5-HT and TH in single neurons can appear after acute stalk section and/or trauma in experimental animals. These findings might represent uptake of exogenous 5-HT or amplified expression of endogenous neurotransmitter, suggesting that plasticity of transmitter phenotype might follow acute surgical and/or endocrine intervention in mature primate brain. Neuroendocrine studies employing the stalk-sectioned primate might thus be radically affected.