Novel mutation of SPG4 gene in a Chinese family with hereditary spastic paraplegia:A case report

BACKGROUND Hereditary spastic paraplegia(HSP)is a group of neurogenetic diseases of the corticospinal tract,accompanied by distinct spasticity and weakness of the lower extremities.Mutations in the spastic paraplegia type 4(SPG4)gene,encoding the spastin protein,are the major cause of the disease.This study reported a Chinese family with HSP caused by a novel mutation of the SPG4 gene.CASE SUMMARY A 44-year-old male was admitted to our hospital for long-term right lower limb weakness,leg stiffness,and unstable walking.His symptoms gradually worsened,while no obvious muscle atrophy in the lower limbs was found.Neurological examinations revealed that the muscle strength of the lower limbs was normal,and knee reflex hyperreflexia and bilateral positive Babinski signs were detected.Members of his family also had the same symptoms.Using mutation analysis,a novel heterozygous duplication mutation,c.1053dupA,p.(Gln352Thrfs*15),was identified in the SPG4 gene in this family.CONCLUSION A Chinese family with HSP had a novel mutation of the SPG4 gene,which is autosomal dominant and inherited as pure HSP.The age of onset,sex distribution,and clinical manifestations of all existing living patients in this family were analyzed.The findings may extend the current knowledge on the existing mutations in the SPG4 gene.

Modulation of valosin-containing protein by Kyoto University Substances(KUS) as a novel therapeutic strategy for ischemic neuronal diseases

Retinal ischemia causes several vision-threatening diseases, including diabetic retinopathy, retinal artery occlusion, and retinal vein occlusion. Intracellular adenosine triphosphate（ATP） depletion and subsequent induced endoplasmic reticulum（ER） stress are proposed to be the underlying mechanisms of ischemic retinal cell death. Recently, we found that a naphthalene derivative can inhibit ATPase activity of valosin-containing protein, universally expressed within various types of cells, including retinal neural cells, with strong cytoprotective activity. Based on the chemical structure, we developed novel valosin-containing protein modulators, Kyoto University Substances（KUSs）, that not only inhibit intracellular ATP depletion, but also ameliorate ER stress. Suppressing ER stress by KUSs is associated with neural cell survival in animal models of several neurodegenerative diseases, such as glaucoma and retinal degeneration. Given that a major pathology of ischemic retinal diseases, other than intracellular ATP depletion, is ER stress-induced cell death, KUSs may provide a novel strategy for cell protection in ischemic conditions. Hence, we investigated the efficacy of KUS121 in a rat model of retinal ischemic injury. Intravitreal injections of KUS121, which is clinically preferable route of drug administration in retinal diseases, significantly suppressed inner retinal thinning and retinal cell death, and maintained visual functions. Valosin-containing protein modulation by KUS is a promising novel therapeutic strategy for ischemic retinal diseases.

Effect of mercuric chloride on membrane-bound enzymes in rat testis

Aim: To study the effect of mercuric chloride on the membrane-bound enzymes. Methods: The effect of mercuric chloride at two different doses, 1 mg/kg (low dose) and 2 mg/kg (high dose), orally for 30 days, was observed on the membrane-bound enzymes in the testis of adult albino rats. Results: Mercuric chloride significantly decreased the body weight and testis weight in the high dose group (P<0.05), but not in the low dose group. The activities of 5' nucleotidase and adenosine triphosphatases were markedly decreased (P<0.01) in the testis of both groups. Alkaline phosphatase and γ-glutamyl transferase activities were significantly increased (P<0.01) in both groups. However, the effect was more pronounced in the high than in the low dose groups. Conclusion: The dose dependent effect of mercuric chloride on these enzymes may affect the membrane characteristics and thereby the fertility of the animal. (Asian J Androl 2002 Dec; 4:309-311)

Ultrastructural and Ultrahistochemical Studies on Rat Hepatocytes under the Effect of Lead Acetate Administration

The present study was carried out to investigate the effect of lead acetate on the ultrastructure of albino rat hepatocytes with special reference to its effect on the mitochondrial and lysosomal activity. Lead acetate was given orally to albino rats at a dose of 1% lead acetate / 100 g body weight, three times / week for one month （Gila）, two months （Glib）, three months （GIIc）. Liver total protein, body / liver weight ratio and cytochrome P-450 value were calculated. Three parts of the liver samples were incubated in media containing adenosine triphosphate, p-nitrophenyl phosphate and 2% glutlraldehyde and prepared for electron microscopy to visualize adenosine triphosphatase, acid phosphatase and the fine structures ofhepatocytes respectively. The main changes of the fine structures was found in the nucleus, as irregularity of the nuclear membrane, clumped heterochromatin and sun radiation of lead inclusion bodies. The cytoplasm was characterized by shortened, dilated rough endoplasmic reticulum, destroyed and hazy mitochondria with increased number of lysosomes with storage secretion. Ultrastructure findings showed hepatocytes damage due to increased hydrolysis enzymes and decreased cytotoxic enzymes （cytochrome P-450） as well as oxidative enzymes that proved the toxic effect of lead according to the duration of exposure.

Effect of Acupuncture on Monoamines and Adenosine Triphosphatase Activityin Lateral Hypothalamic Area of ObeseRats

Objective: To study the mechanism of acupuncture in treating simple obesity. Methods: Central nerve push-pull perfusion and biochemical technique were used to observe the effect of acupuncture on the obese parameters, changes of monoamine transmitters and activity of ATPase in the lateral hypothalamic area (LHA) of obese rats. Results: Noradrenaline (NA) level in LHA of obese rats was higher but serotonin (5-HT) level and ATPase activity were lower than those in normal rats. After acupuncture treatment, in the same time of reducing body weight, NA level in LHA of rats was reduced, and 5-HT level and ATPase activity in it were increased.(P<0.05 and P<0.01). Conclusion:The effective regulation on LHA of obese rats is possibly one of the key factors in anti-obesity effect of acupuncture.

Effects of three kinds of head acupuncture therapies on regulation of brain microenvironment and rehabilitation of nerve function in rats with cerebral palsy

OBJECTIVE:To compare and observe the effects of three kinds of cephalic acupuncture therapies commonly used in the clinic on promoting nerve function rehabilitation in the brain microenvironment of rats with cerebral palsy.METHODS:A negative control group,positive control group,and three cephalic acupuncture groups based on the administration of three cephalic acupuncture therapies were established.Ten experimental rats were selected from each group at 1,2,and 3 weeks after modeling.Neuromotor function after treatment was rated according to the Basso,Beattie,and Bresnahan method.White matter fiber bundles were evaluated by head diffusion tensor imaging.The expression levels of neuron-specific enolase,microtubule-associated protein 2,and myelin basic protein in the brain tissue extract were detected by Western blot analysis and the activities of ATPases were determined using a fixed phosphorus method.RESULTS:The pathological changes in brain tissue were restored and motor function scores were increased in the mice in each cephalic acupuncture group,and the expression of neuronal growth-related proteins in the brain tissue extract was significantly increased.Additionally,the activities of ATPases in the lesion area were significant enhanced(P<0.05).Diffusion tensor imaging revealed that the white matter fiber bundles of mice in each cephalic acupuncture group gradually increased and recovered.The nervous system structure was significantly improved.CONCLUSIONS:All three acupuncture methods promoted the rehabilitation of nerve function damaged by cerebral palsy.These effects are likely related to the improved expression of nerve growth-related proteins,enhancement of ATPase activities,and regulation of the brain microenvironment.

Protective effects of penehyclidine hydrochloride on acute lung injury caused by severe dichlorvos poisoning in swine

Background Organophosphate poisoning is an important health problem in developing countries which causes death mainly by inducing acute lung injury. In this study, we examined the effects of penehyclidine hydrochloride （PHC）, a selective M-receptor inhibitor, on dichlorvos-induced acute lung injury in swine. Methods Twenty-two female swines were randomly divided into control （n=5）, dichlorvos （n=6）, atropine （n=6）, and PHC （n=5） groups. Hemodynamic data, extravascular lung water index （EVLWI）, and pulmonary vascular permeability index （PVPI） were monitored; blood gas analysis and acetylcholinesterase （AchE） levels were measured. PaO2/FiO2, cardiac index （CI）, and pulmonary vascular resistance indices （PVRI） were calculated. At termination of the study, pulmonary tissue was collected for ATPase activity determination and wet to dry weight ratio （W/D） testing 6 hours post-poisoning. TUNEL assay, and Bax, Bcl-2, and caspase-3 expression were applied to pulmonary tissue, and histopathology was observed. Results After poisoning, PHC markedly decreased PVRI, increased CI more effectively than atropine. Anticholinergic treatment reduced W/D, apoptosis index （AI）, and mitigated injury to the structure of lung; however, PHC reduced AI and caspase-3 expression and improved Bcl-2/Bax more effectively than atropine. Atropine and PHC improved ATPase activities; a significant difference between groups was observed in Ca2＋-ATPase activity, but not Na＋-K＋-ATPase activity. Conclusions The PHC group showed mild impairment in pathology, less apoptotic cells, and little impact on cardiac function compared with the atropine group in dichlorvos-induced acute lung injury.

Effect of Moxibustion on the Motility, Absorption and Activities of ATPase in Small Intestine of Spleen-deficiency Rats

Objective： To observe the effect of moxibustion on intestinal propulsion rate, content of serum D-xylose, content of adenosine triphosphate （ATP） and activities of adenosine triphosphatases （ATPase） in jejunum tissues of spleen-deficiency rats, and to explore the effect of moxibustion on the production of ATP and activities of membrane proteins, for analyzing the mechanism of moxibustion in reinforcing the spleen-stomach. Methods： Forty healthy Sprague Dawley rats were randomly divided into four groups, namely group A （blank control group）, group B （model control group）, group C （moxibustion group） and group D （herbs group）. The animal model of spleen-deficiency was established by intragastric administration with Da Huang （Radix et Rhizoma Rhei） infusion. The rats in group C received moxibustion at Zusanli （ST 36）, Zhongwan （CV 124 Guanyuan （CV 4）, Pishu （BL 20） and Weishu （BL 21）. The rats in group D received intragastric administration with 5i Jun Zi decoction. The intestinal propulsion was measured by toner method, and the content of D-xylose in serum was detected by phloroglucinol method. Colorimetry method was used to detect the content of ATP and activities of Na^＋-K^＋-ATPase and Ca^2＋-Mg^2＋-ATPase in jejunum tissues. Results： Compared with group A, the intestinal propulsion rate, content of D-xylose in serum, content of ATP, activities of Na^＋-K^＋-ATPase and Ca^2＋-Mg^2＋-ATP in jejunum tissues decreased significantly in group B （P〈0.05 or P〈O.O1）. Compared with group B, the intestinal propulsion rate, content of D-xylose in serum, content of ATP, activities of Na^＋-K^＋-ATPase and Ca^2＋-Mg^2＋-ATPase in jejunum tissues increased significantly in group C and D （P〈0.05 or P〈0.01）, while there were no significant differences between group C and group D. Conclusion： Moxibustion can increase the intestinal propulsion rate, content of D-xylose in serum and ATP in jejunum tissues, as well as the activities of Na^-K＋-ATPase and Ca^2＋-Mg^2＋-ATPase of spleen-deficiency rats, which suggests that moxibustion can enhance the motility and absorption functions of small intestine by promoting the production of ATP in intestinal epithelial cells, increasing the content of membrane proteins, and improving transmembrane transport.