Induction of GADD45α protects M17 neuroblastoma cells against MPP*.

Growth arrest and DNA-damage-inducible protein 45α (GADD45α) is an important member of the family of growth arrest and DNA damage-inducible (GADD) proteins. The expression patterns and possible roles of GADD45α in Parkinson's disease (PD) are so far less understood. In this study, we found that 1-methyl-4-phenylpyridinium (MPP+) treatment up-regulates the expression of GADD45α in both a time-dependent manner and a dose-dependent manner in human dopamine neuroblastoma M17 cells. The up-regulation of GADD45α was abolished by pretreatment with the c-Jun N-terminal kinases (JNK) inhibitor SP600125 but not the p38 specific inhibitor SB203580. Further study revealed that c-Jun silencing abolished the effects of MPP+ on the expression of GADD45α. Important, ChIP studies verified the ability of c-Jun to bind to the GADD45 promoter. In addition, we found that inhibition of GADD45α by small RNA interference exacerbates the impaired cell viability, LDH release, and apoptosis induced by MPP+. Correspondingly, silence of GADD45 exacerbated Caspase-3 activation induced by MPP+. These data suggested a neuroprotective effect of GADD45α against MPP+ neurotoxicity.

[Roles of bcl-2/bax expression and oligodendrocyte apoptosis in the pathogenesis of heroin-induced spongiform leucoencephalopathy].

OBJECTIVE: To investigate the role of oligodendrocyte apoptosis under the regulation of the bcl-2/bax protein expression in brain white matter in the pathogenesis of heroin-induced spongiform leucoencephalopathy (HSLE). METHODS: Samples of frontal lobe, cerebellum, and corpus callosum were obtained from the brains during autopsy of 4 HSLE cases and 5 normal controls and underwent light microscopy and electron microscopy. Immunocytochemistry was used to detect the expression of myelin basic protein (MBP), caspase-3, bcl-2 protein, and bax protein. RESULTS: Widespread demyelination was seen in the white matter of the frontal lobe, cerebellum and corpus callosum of the HSLE cases, most severely in the cerebellum. The levels of caspase-3 and bax expression of the HSLE group were significantly higher than those of the control group (both P <0.05) , however, the bcl-2 level of the HSLE group was no significantly different from that of the control group (P > 0.05). CONCLUSION: Widespread demyelination in the white matter is a prevailed pathological change of HSLE. Oligodendrocyte apoptosis under induced by the decrease of bcl-2/bax ratio may contribute to the pathogenesis.

[Roles of cyclooxygenase 2/2', 3'-cyclic nucleotide3' phosphohydrolase in the oligodendrocyte apoptosis in heroin-induced spongiform leucoencephalopathy].

OBJECTIVE: To investigate the regulation of cyclooxygenase (Cox)-2/2', 3'-cyclic nucleotide3' phosphohydrolase (CNPase) on the oligodendrocyte apoptosis in the pathogenesis of the heroin-induced spongiform leucoencephalopathy (HSLE). METHODS: Samples of frontal lobe, cerebellum, and corpus callosum were obtained from the brains during autopsy of 4 HSLE patients and 5 patients who died of diseases other than cerebral diseases (controls) and underwent light microscopy and electron microscopy. Immunocytochemistry was carried out to detect the expression of myelin basic protein (MBP), caspase-3, COX-2, and CNPase protein. Apoptosis was examined by TUNEL staining. RESULTS: Widespread demyelination was seen in the white matter of the frontal lobe, cerebellum, and corpus callosum of the HSLE cases, most severely in cerebellum. In he HSLE group, the levels of caspase-3 and COX-2 expression were significantly higher, and the level of CNPase was significantly lower than those of the control group (all P < 0.05). CONCLUSION: Widespread demyelination in the white matter is a prevailing pathological change of HSLE. Oligodendrocyte apoptosis is one of the causes of HSLE. The upregulation of COX-2 and downregulation of CNPase may contribute to the pathogenesis.

[Relationship between polymorphisms of interleukin 10 promoter and serum levels of lipoprotein in the Chinese Han population].

OBJECTIVE: To study the relationship between polymorphisms of interleukin 10 (IL10QX) promoter and serum levels of lipoprotein in the healthy Chinese Han population. METHODS: PCR restriction fragment length polymorphism assay was used to detect the distribution of genotypes of IL10 -592,-819,-1082 in 200 healthy Chinese Han subjects. Serum levels of total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and very low-density lipoprotein (VLDL) in all subjects were measured to analyze the relationship with the polymorphisms of IL10 promoter. RESULTS: Comparing with AA genotype, the group with GA genotype at IL10 promoter -1082 position had a significant elevation of serum HDL-C level [(1.514+/-0.501) mmol/L vs. (1.261+/-0.346) mmol/L, t=-2.225, P=0.028] and a lower serum TG level[(1.701+/-1.836) mmol/L vs. (0.981+/-0.314) mmol/L,Z=-2.096,P=0.036]. The TG, TC, HDL-C, LDL-C and VLDL levels did not show any statistically significant differences among different genotypes (CC, AA, CA) of the IL10 -592, as well as the genotypes (TT, TA, AA) ofIL10 -819 (P>0.05). CONCLUSION: The results suggest that in the Chinese Han population, the polymorphism at position -1082 in the promoter region of IL10 gene may be associated with the serum HDL-C level and TG level.

Cloning and sequencing of junction fragment with exons 45-54 deletion of dystrophin gene.

OBJECTIVE: To study the mechanisms of dystrophin gene deletion, the junction fragment with exons 45-54 deletion were cloned and sequenced. METHODS: A Duchenne muscular dystrophy (DMD) patient with exons 45-54 deletion has been substantiated by PCR amplification of the exons. Then we used a PCR-based genome-walking method for localizing the breakpoints in introns 44 and 54. At last, the deletion-junction fragment was directly amplified by PCR approach with forward and reverse primers annealing to a DNA sequence as close as possible to the breakpoints in introns 45 and 54. The sequencing result of the deletion-junction fragment was compared with the normal intronic sequences. RESULTS: A total of 2716 bp sequence containing the junction fragment was obtained. The 5' breakpoint was located in LINE/L1 element of intron 44 and close to a matrix attachment region (MAR). The 3' breakpoint was located in the minor potential MAR with topoisomerase II cleavage sites around. Beside the 3' breakpoint there was a 6 bp palindromic sequence. A 4 bp microhomologous sequence (AGAG) was in the joint of the deletion-junction fragment. CONCLUSION: The nonhomologous recombination caused by L1 repeated element, topoisomerase II cleavage sites, MARs and the nonhomologous end joining of microhomologous sequence may be the important factors in this huge gene fragment deletion.

Diminution of toxic copper accumulation in toxic milk mice modeling Wilson disease by embryonic hepatocyte intrasplenic transplantation.

AIM: To observe the therapeutic effect of intrasplenic transplantation with embryonic hepatocytes on amelioration of hereditary copper accumulation in toxic milk (TX) mouse modeling Wilson disease. METHODS: Donor hepatocytes were harvested from 14-d fetal liver of a pregnant homogeneous DL mouse. These cells were successively cultured, labeled with fluorescein dye Hoechst 33342 for 24 h, and sequentially infused into the spleen parenchyma of the recipient TX mice. No host immunosuppression measures were taken. Two and four weeks after transplantation, the recipients were killed for routine histologic investigation and immunohistochemistry study up to 4 wk after transplantation. The serum copper and ceruloplasmin concentrations of the recipient mice were determined by graphite furnace atomic absorption spectroscopy. RESULTS: In the following 2nd and 4th wk after transplantation, the donor hepatocytes could be visualized in the livers of 47.3% recipients. The serum ceruloplasmin and copper concentrations increased by 1.6-fold after 2 wk and 2.0-fold times after 4 wk respectively, which ultimately rose from about 30% of the normal level to nearly 60% (P<0.01). The hepatic copper concentration decreased 7.2%, 4 wk after transplantation. Pathologic examination showed that there were many actively proliferative hepatocyte precursor cells with specific embryonic hepatocyte marker AFP migrated into hepatic sinusoids of the recipients. A large number of cells carrying hepatocytes marker and albumin were observed in the recipient spleen tissues. CONCLUSION: Embryonic hepatocytes are capable of differentiating into mature hepatocytes in vivo. After transplantation, the hereditary abnormalities of copper metabolism in TX mice could be corrected partially by intrasplenic transplantation of homogeneous embryonic hepatocytes.

[Effects of thyroid hormone on cognitive function in rats with chronic cerebral ischemia].

OBJECTIVE: To study the characteristics of cognitive dysfunction in rats with chronic cerebral ischemia and the effects of thyroid hormone on the rats' cognitive function. METHODS: Thirty-one male SD rats were randomly allocated into normal control group (n=12), operation group (with bilateral carotid artery ligation and examined 5 weeks later, n=6), acute phase treatment group (APT, with bilateral carotid artery ligation and intragastric administration of thyroid hormone at 20 mg once daily for 5 weeks starting from the day of operation, n=7) and chronic phase treatment group (CPT, with the operation and thyroid hormone administration in an identical manner started from the sixth week following the operation, n=6). Morris water maze test was performed at the end of experiment. One-way ANOVA was used to estimate the differences in the learning and memory functions of the rats using SPSS10.0 for Windows. RESULTS: The average latent period of the operation group was significantly prolonged in comparison with that of the other groups (P<0.05) in spatial orientation test. The probe time (time spent in the quadrant where the platform was once situated) of normal control was much shorter than those of the operation, APT and CPT groups in spatial probe test (P<0.05), and the operation group had the poorest score. The average latent period of the operation and CPT was longer than that of the other groups (P<0.05) in working-memory task (P<0.05), and the operation group again had the poorest score. CONCLUSION: Spatial cognitive function is totally damaged in rats with chronic cerebral ischemia, and learning can not induce the formation long-term memory because of short-term memory damage. Thyroid hormone may lessen but can not fully repair the damage of the cognitive dysfunction resulting from chronic cerebral ischemia, and early intervention with thyroid hormone may be beneficial for chronic cerebral ischemia.

[Expression of glucose transporter 1 in the blood-brain barrier around hematoma in rats with cerebral hemorrhage].

OBJECTIVE: To observe the expression of glucose transporter 1 (GLUT1) in the blood-brain barrier around the hematoma in rats with cerebral hemorrhage. METHOD: Brain tissue around the hematoma in rats with cerebral hemorrhage was prepared for GLUT1 detection by immunohistochemistry. RESULT: The expression of GLUT1 upregulated within 48 h after cerebral hemorrhage and downregulated afterwards. CONCLUSION: Protective reaction of the blood-brain barrier can be initiated to ensure glucose supply to the neurons against the injury by cerebral hemorrhage.

[Clinical analysis of 73 cases of hypoglycemia with brain dysfunctions].

OBJECTIVE: To identify the clinical features and pathogenesis of hypoglycemia with brain dysfunctions as the main manifestations. METHOD: A retrospective analysis of 73 cases with brain dysfunctions caused by hypoglycemia was performed. RESULT: Hypoglycemic brain dysfunctions were mainly caused by poor control of the dosage of hypoglycemic agents in diabetic patients, and the major clinical manifestations included coma, hemiparalysis, epilepsy and mental disorders. The pathogenesis was complicated, possibly related to a variety of factors such as rapid declination of blood sugar level, aging, cerebral arteriosclerosis, and hypoglycemic cerebral vasospasm or selective nerve injury. CONCLUSIONS: A blood sugar test should be performed for any patients with brain dysfunctions of unidentified causes for early diagnosis and treatment. In addition, diabetic patients should take hypoglycemic agent adequately. Regular monitoring of blood sugar level is key to the prevention of hypoglycemia.

Single photon emission computerized tomography of spongiform leukoencephalopathy heroin addicts: analysis of 10 cases.

OBJECTIVE: To investigate the changes in cerebral circulation in heroin addicts with pongiform leukoencephalopathy. METHODS: Single photon emission computerized tomography (SPECT) was performed in 10 such patients. RESULTS: Regional cerebral blood flow (rCBF) in the involved white matter of bilateral cerebral and cerebellar hemispheres was obviously reduced. rCBF of temporal lobes, parietal lobes, cerebellar hemispheres and basal ganglion were reduced in varying degrees. CONCLUSION: As demonstrated by the result of SPECT, rCBF in the white matter of bilateral cerebral and cerebellar hemispheres was reduced with partial involvement of the gray matter of heroin addicts with spongiform leukoencephalopathy.

[Neuroprotective effect of monoamine oxidase monoclonal antibody against vasogenic brain edema in rats].

OBJECTIVE: To observe the therapeutic effect of monoamine oxidase (MAO) monoclonal antibody (mAb) for vasogenic brain edema (VBE) in rats. METHODS: A total of 75 Wistar rats were randomized into non-edema, non-treated edema, saline-treated edema, mannitol-treated edema and MAO mAb-treated groups. Rat models of VBE were established by intraperitoneal injection of phenylephrine in the latter 4 groups. Brain water content in the gray and white matter was measured respectively with a moisture analyzer, and the permeability of the blood-brain barrier (BBB) determined by Evan's blue (EB) extravasation method. RESULTS: MAO mAb administration significantly reduced the brain water content in the gray and white matter as well as the permeability of BBB (P<0.01), Which was especially effective for the white matter, producing results comparable with those of the non-edema group (P>0.05). MAO mAb markedly alleviated brain edema, with better dehydrating effect on the white matter than mannitol (P<0.01), which reduced the water content of the brain gray and white matter undiscriminatingly and showed poor effect on the permeability of BBB. CONCLUSION: The pathogenesis of BBB permeability changes in VBE is related to the activity of monoamine oxidase, and MAO mAb has selective therapeutic effect on VBE, which is especially obvious on brain white matter.

[Clinical and pathological investigation of polymyositis and dermatomyositis: report of 83 cases].

OBJECTIVE: To study the clinical and pathological characteristics and treatment of polymyositis (PM) and dermatomyositis (DM). METHODS: Eighty-three DM/PM cases were reviewed and analyzed. RESULTS: The main clinical presentations of PM/DM included amyosthenia, muscular tenderness, elevation of serum enzymes, accompanied by abnormal electromyography and muscular pathology. A total efficacy rate of 86.7% was achieved after treatment with corticosteroid and immuno- depressants. CONCLUSION: The different clinical and pathological presentations of various types of PM/DM suggest different pathogenesis between DM and PM. Nerve damage is a part of the systemic damages due to the disease, the prognosis of which is related to its classification and then timing of the treatment. Early effective treatment can improve the prognosis.

C242T p22phox gene polymorphism in the population of Han nationality in Guangdong province.

This study aimed to investigate the gene polymorphism of the C242T p22phox in the population of Han nationality in Guangdong Province by randomly selecting 122 unrelated healthy candidates examined by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). It was found that the C and T allele frequencies were 0.934 and 0.066, and CC and CT+TT genotype frequencies were 0.877 and 0.123, respectively, in the population of Han nationality in Guangdong Province, suggesting the presence of C242T p22phox gene polymorphism with significant racial differences.

[Meningoencephalitis caused by Angiostrongylus cantonensis: report of 1 case].

One rare case of meningoencephalitis caused by Angiostrongylus cantonensis is reported. The development of the disease, clinical features, accessory examinations, and the subsequent treatment were described and analyzed briefly.

Detection of an unkown peak at 1.2 ppm in proton magnetic resonance spectra of cats with cerebral ischemic necrosis.

OBJECTIVE: To confirm an unknown peak (Pu) in proton magnetic resonance spectra ((1)H-MRS) in cats with permanent focal cerebral ischemia, and surmise its potential value of application. METHODS: After focal cerebral ischemia, diffusion-weighted imaging (DWI) was used to identify regions of ischemia for (1)H-MRS voxel localization with the assistance of stereotaxic atlas of cat brain. (1)H-MRS was used to monitor the progression of focal cerebral ischemia in 6 cats over a period of 7 d following middle cerebral artery occlusion (MCAO). The changes in lactate, N-acetyl-aspartate (NAA), choline, creatine, and Pu were observed. RESULTS: In the involved regions, lactate was elevated almost immediately after the onset of cerebral ischemia, and NAA declined within several hours of acute infarction. The Pu at 1.2 ppm was persistently detected in the affected cerebral areas 2 to 7 d after MCAO. CONCLUSION: Pu has a close relationship with cerebral ischemia necrosis and may be a intrinsic production of the necrotic tissue, which can be utilized as a specific marker and therefore has important clinical diagnostic value.

[Surgical procedure improvement to produce rat ischemia-reperfusion injury model with intraluminal suture].

OBJECTIVE: To describe a modified surgical approach to produce rat ischemia-reperfusion injury model with intraluminal suture. METHODS: After exposure and ligation of the common carotid artery (CCA), the left external carotid artery and pterygopalatine artery were opened in which a 3-0 nylon suture was introduced intraluminally from the distal end of the ligature of the CCA using a scalp needle. Reperfusion injury was induced by withdrawal of the suture. RESULTS: The length of the suture was about 20.0+/-1.8 mm and the success rate of model establishment was nearly 70%. The rats developed typical symptoms and pathological manifestations after the surgery. CONCLUSION: This modified surgical approach is simple for model establishment and does not require special microsurgical skills to ensure the high success rate.

[Evaluation of steal phenomena by measuring the steal index using transcranial Doppler ultrasound in intracranial arteriovenous malformation].

OBJECTIVE: The authors previously deduced a formula for calculating the steal index (SI) for evaluation of the steal phenomena during the development of the collateral circulation resulting from unilateral carotid artery occlusion or stenosis. In this study the authors examine the application of SI in evaluating the steal phenomena of cerebral blood flow in cases of intracranial arteriovenous malformation (AVM). METHODS: The clinical data of 16 cases of AVM confirmed by both digital subtraction angiography (DSA) and transcranial Doppler (TCD) ultrasound were analyzed retrospectively. The ratio of the mean blood velocity in the direct or indirect feeding artery (Vmfv) for the area containing AVM to the mean blood velocity in the neighboring feeding artery (Vmna), Vmfv/Vmna, was compared with SI of the corresponding arteries (SI=1-PI(1)/PI(2), where PI(1) is the pulsatility index of the feeding artery and PI the pulsatility index of the neighboring feeding artery), and the correlation between SI and the size of AVM was analyzed statistically. RESULTS: The Vmfv/Vmna of (2)direct and indirect feeding arteries for AVM both exhibited positive correlation with SI (r=0.62, P<0.05, n=16; r=0.53, P<0.01, n=27), and SI appeared to be in positive correlation with the size of AVM, but which failed to be supported by statistical analysis (r=0.48, P>0.05, n=12). CONCLUSION: The sizes, types and development of the vascular bed of AVM can be evaluatea by analysis of SI derived from TCD in combination with DSA.

Deduction of regional cerebral blood flow loss index after stenosis in cerebral artery.

OBJECTIVE: To study the calculation of cerebral blood flow loss ratio after cerebral artery stenosis using the data obtained from transcranial Doppler sonography (TCD), by deducing regional cerebral blood flow loss index (rCBFLI). METHODS AND RESULTS: The data of TCD performed in 31 cases of unilateral stenosis in the middle cerebral artery (MCA) were collected and analyzed for deducing the formula for calculating the MCA stenosis index (STI): STI=1 (Vm(0)/Vm(1)) x (PI(1)/PI(0)), where Vm(0) is the mean normal velocity of MCA blood flow (derived from the measurements in 908 normal subjects at varied ages receiving TCD) and Vm(1) the mean velocity at the stenosed MCA, and PI(0) and PI(1) stand for the pulsatility index before and after stenosis. The ratio of PI(1) to PI(0) is likely to be equivalent to the ratio of Q(1) to Q(0) (Q(0) and Q(1) represent the volume of blood flow in the MCA in normal condition and after stenosis, respectively), thus the equivalence can be extended as rCBFLI=(1 Q(1)/Q(0)) x 100%=(1 PI(1)/PI(0)) x 100%. rCBFLI after MCA stenosis was calculated in 31 cases accordingly, and by means of correlation analysis, we found positive correlation between rCBFLI and Vm(1) (r=0.76, P < 0.001) and between Vm(1) and STI (r=0.85, P < 0.001). In the same way, rCBFLI of 55 sides with MCA stenosis in 43 cases were analyzed, and positive correlation between rCBFLI and Vm (r=0.76, P < 0.001) and between rCBFLI and STI (r=0.83, P < 0.001) was found. CONCLUSION: rCBFLI can be used to evaluate the degree of regional cerebral blood flow loss due to artery stenosis, and the combination of rCBFLI and STI may provide an insight into the changes of cerebral hemodynamics in this pathologic condition.

[Glucocortioid treatment for heroin-induced spongiform leucoencephalopathy: a clinical controlled study].

OBJECTIVE: To evaluate the therapeutic effects of the glucocorticoid on heroin-induced spongiform leucoencephalopathy. METHODS: Twenty cases of heroin-induced spongiform leucoencephalopathy were randomly divided into the control group and the treating group with equal number. In the control group, the treatment was constituted by oral administration of vitamin B and coenzyme Q10 in a course of 1 month. In glucocorticoid treatment group, glucocorticoid (20 mg/d) for 10 d were given in addition to vitamin B and coenzyme Q10, and the dose of the glucocorticoid was gradually decreased afterwards. General observation and statistical analysis of function scores were performed in both groups before and 1, 6, 12 months after the treatment respectively. RESULTS: No significant difference in function scores was observed between the 2 groups, while the results of observation before and after the treatment were significantly different (P<0.05). The most significant difference occurred when comparing the observations made 1 month and 6 months respectively after treatment (P<0.001). CONCLUSION: Glucocorticoid has no obvious therapeutic effect on heroin-induced spongiform leucoencephalopathy, and rapid clinical recovery occurs within the initial 6 months of the treatment.

[A rat model of focal lymph encephalopathy established by partial ligation of the cerebral superficial artery].

OBJECTIVE: To establish a rat model of focal lymphatic encephalopathy by partial ligation of the cerebral superficial artery for observation of the changes of Virchow-Robin spaces (VRS). METHODS: Thirty male SD rats were randomized into 3 groups (n=10), including two model groups and a sham-operated group. The rats in the model groups were subjected to partial ligation of the cerebral superficial arteries under EEG monitoring to induce focal lymphatic encephalopathy, and those in the sham-operated group underwent only dissociation of the cerebral superficial artery without ligation. The rats in the two model groups were executed at 24 and 48 h, and those in the sham-operated group at 48 h following the operation, respectively. Frozen sections of the brain tissues were prepared for microscopic morphological observation and quantitative analysis of the VRS using HE staining and an image analysis system, respectively. RESULTS: EEG remained normal during the operations suggesting intact brain function. Partial ligation of the cerebral superficial arteries resulted in obvious dilation of the VRS in the cerebral cortex and subcortical medulla, and the tissues around the dilated VRSs appeared pale and structurally loosened. The two model groups showed significantly enlarged VRS areas as compared to the sham-operated group (P<0.01), but no significant differences were found in the mean VRS areas between the two model groups. CONCLUSION: Partial dilation of the cerebral superficial artery is effective and convenient to induce focal lymphatic encephalopathy in rats, and this model can be ideal for studying focal cerebral lymph circulation.