OBJECTIVE: To explore the clinical, electrophysiological and imaging features of a patient with Krabbe disease caused by GALC mutation. METHODS: A comprehensive analysis including clinical investigation and genetic testing was carried out. RESULTS: The patient presented with peripheral neuropathy with electrophysiological anomaly suggestive of asymmetric demyelinating neuropathy. Brain imaging revealed leukoencephalopathy. Genetic analysis has identified compound heterozygous mutations in exons 5 and 11 of the GALC gene, namely c.461C>A and c.1244G>A. CONCLUSION Krabbe disease is a group of disorders featuring substantial phenotypic heterogeneity. Genetic and enzyme testing has become indispensable for accurate diagnosis for this disease.
DNA Mutational Analysis ; Galactosylceramidase ; genetics ; Genetic Testing ; Humans ; Leukodystrophy, Globoid Cell ; complications ; genetics ; Mutation ; Peripheral Nervous System Diseases ; etiology

The purpose of this study was to study the role of neurofilament (NF) mRNA and calpain in NF reduction of acrylamide (ACR) neuropathy. Male Wistar adult rats were injected i.p. every other day with ACR (20 mg/kg·bW or 40 mg/kg·bW) for 8 weeks. NF mRNA expression was detected using RT-PCR and the calpain concentration was determined using western blot analysis. The NF mRNA expression significantly decreased while the level of m-calpain and μ-calpain significantly increased in two ACR-treated rats groups regardless of the ACR dose. The light NF (NF-L) protein expression was significantly correlated with NF-L mRNA expression. Combined with previous data, the concentrations of three NF subunits were negatively correlated with the calpain levels. These findings suggest that NF-L mRNA and calpain mediated the reduction in NF of ACR neuropathy.
Acrylamide ; toxicity ; Animals ; Calpain ; metabolism ; Gene Expression Regulation ; drug effects ; Intermediate Filaments ; genetics ; Male ; Peripheral Nervous System Diseases ; chemically induced ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats

PURPOSE: Floppy infant, or congenital hypotonia, is caused by various diseases, such as genomic disorders, diseases involving the central or peripheral nervous system, musculoskeletal diseases, and metabolic disorders. We describe here the clinical aspects and the final diagnosis of infants with hypotonia recently diagnosed in a single, tertiary-care hospital in Korea. METHODS: All of the infants evaluated for generalized hypotonia between 2008 and 2012 at Asan Medical Center Children's Hospital were included in our study. The demographic data, physical examination upon initial presentation, the diagnostic tests and results, and the final diagnosis were retrospectively reviewed. RESULTS: A total of 128 infants (68 males, 60 females) were included in the study, and the mean patient age at the time of the diagnosis of hypotonia was 4.8 months. Etiological diagnosis was possible in 80 (62.5%) of the 128 patients, and 57 (44.5%) patients were confirmed by genetic testing. Fifteen patients (11.7%) were categorized as having central nervous system disorders, and 34 (26.6%) patients were diagnosed as having other genomic disorders such as Prader-Willi syndrome (n=17). Disease involving muscle and the peripheral nervous system was detected in 16 (12.5%) patients. Five patients were diagnosed with other skeletal disorders, and metabolic disease was detected in 10 (7.8%) patients. CONCLUSION: With the recent advances in diagnostic tools, including genetic testing, many of the patients with hypotonia can be correctly diagnosed. These data can give practical clues regarding the optimal diagnostic approaches for treating floppy infants in the clinics.
Central Nervous System Diseases ; Chungcheongnam-do ; Diagnosis ; Diagnostic Tests, Routine ; Genetic Testing ; Genetics ; Humans ; Infant* ; Korea ; Male ; Metabolic Diseases ; Muscle Hypotonia ; Musculoskeletal Diseases ; Peripheral Nervous System ; Physical Examination ; Prader-Willi Syndrome ; Retrospective Studies

OBJECTIVE: Distal hereditary motor neuropathy (dHMN) comprises a heterogeneous group of inherited disorders associated with neurodegeneration of motor nerves and neurons, mainly charac-terized by progressive atrophy and weakness of distal muscle without clinical or electrophysiological sensory abnormalities. To improve the recognition and diagnosis of the disease, we summarized the clinical manifestations, electrophysiological, pathological, and genetic characteristics in eight patients with dHMN. METHODS: Eight probands from different families diagnosed with dHMN were recruited in this study between June 2018 and April 2019 at Peking University People's Hospital. Eight patients underwent complete neurological examination and standard electrophysiological examinations. The clinical criteria were consistent with the patients presenting with a pure motor neuropathy with no sensory changes on electrophysiology. The detailed clinical symptoms, neurophysiological examinations, pathological features and gene mutations were analyzed retrospectively. Genetic testing was performed on the eight patients using targeted next-generation sequencing panel for inherited neuromuscular disorder and was combined with segregation analysis. RESULTS: The age of onset ranged between 11 and 64 years (median 39.5 years) in our dHMN patients. All the cases showed a slowly progressive disease course, mainly characterized by distal limb muscle weakness and atrophy. The motor nerve conduction revealed decreased compound muscle action potential amplitude and velocity, while the sensory nerve conduction velocities and action potentials were not affected. Needle electromyography indicated neurogenic chronic denervation in all patients. Muscle biopsy performed in two patients demonstrated neurogenic skeletal muscle damage. Sural nerve biopsy was performed in one patient, Semithin sections shows relatively normal density and structure of large myelinated fibers, except very few fibers with thin myelin sheaths, which suggested very mild sensory nerve involvement. Eight different genes known to be associated with dHMN were identified in the patients by next-generation sequencing, pathogenic dHMN mutations were identified in three genes, and the detection rate of confirmed genetic diagnosis of dHMN was 37.5% (3/8). Whereas five variants of uncertain significance (VUS) were identified, among which two novel variants co-segregated the phenotype. CONCLUSION dHMN is a group of inherited peripheral neuropathies with great clinical and genetic heterogeneity. Next-generation sequencing is widely used to discover pathogenic genes in patients with dHMN, but more than half of the patients still remain genetically unknown.
Adolescent ; Adult ; Child ; Hereditary Sensory and Motor Neuropathy/genetics* ; Humans ; Middle Aged ; Mutation ; Peripheral Nervous System Diseases ; Phenotype ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate an effective treatment of peripheral nerve injuries by means of gene transference.

METHODS48 adult Wister rats were divided evenly into 3 groups. A 10 mm sciatic nerve gap was created and bridged with a silicone chamber. The silicone chamber was filled with glial cell-line derived neurotrophic factor(GDNF) gene modified Schwann cells(SCs) (group 1), the normal SCs(group 2) and nothing(the control). At 4, 8, 12, and 16 weeks after the operation, the general and histological observations, the electromyographic and immunohistochemical examinations were performed to the regenerated nerves.

RESULTSThe GDNF-SCs group was significantly better than the SCs and the control groups in nerve conduction velocity, the number and density of reinnervation, the area of regenerated nerve and the thickness of myelin sheath of the regenerated nerves.

CONCLUSIONGDNF gene modified SCs secrete higher levels of neurotrophic factors for a prolonged time, which are more effective in peripheral nerve repair than the normal SCs.

Animals ; Female ; Glial Cell Line-Derived Neurotrophic Factor ; Immunohistochemistry ; Male ; Nerve Growth Factors ; analysis ; genetics ; physiology ; Nerve Regeneration ; physiology ; Peripheral Nerves ; chemistry ; physiopathology ; Peripheral Nervous System Diseases ; surgery ; Rats ; Rats, Wistar ; Schwann Cells ; metabolism ; transplantation

OBJECTIVETo observe the effects and duration of electroacupuncture on the mechanical pain threshold induced by paclitaxel and explore its analgesic mechanism.

METHODSSixty-four C57BL/6J male mice were randomly divided into 4 groups, a normal+sham EA group, a normal+EA group, a medicine+sham EA(Med+ sham EA) group, a medicine + EA (Med + EA) group, 16 cases in each group. The model of chemotherapy-induced peripheral neuropathy was established with paclitaxel intraperitoneal injection on the 1st, 3rd, 5th, 7th day in the Med + sham EA group and the Med + EA group. EA of 30 min was used on bilateral "Zusanli (ST 36)" on the 9th, 11th, 13th, 16th, 18th, 20th, 23rd, 25th, 27th, 30th day in the EA groups, 2 Hz/100 Hz and 1~ 1.5 mA. Acupuncture was applied on the same acupoint at the same times in the sham EA groups. Mechanical pain thresholds were tested by VonFrey before and after model establishment, namely on the 8th, 14th; 21st and, 28th day. The heart blood of 8 mice was drawn quickly to collect serum in every group on the 31st day, and the contents of tumor necrosis factor α (TNF-α), interleukin-1α (IL-1α), interleukin-1β (IL-1β) in proinflammatory cytokine were examined by ELISA. Mechanical pain thresholds were tested by VonFrey for the rest 8 mice of each group until there was no apparent difference in the two paclitaxel groups once a week,namely on the 35th, 42nd, 49th day.

RESULTSThe pain thresholds of each group were not statistically different before model establishment (P > 0.05). After model establishment (on the 8th day), thresholds of the paclitaxel groups were lower than those of the normal groups (all P < 0.05). After EA, the mechanical pain thresholds of the Med + EA group were higher than those of the Med + sham EA group at all the time points, and there was statistical difference on the 14th, 21st and 28th day (all P < 0.05). The analgesic effect was lasting to the 49th day. The contents of TNF-α, IL-1α, IL-1β of the Med + EA group were decreased than those of the Med+sham EA group in different degree, with statistical significance of IL-1α (P < 0.05).

CONCLUSIONEA can effectively treat paclitaxel-induced peripheral neuropathy,and the analgesic mechanism is probably related to decreasing the proinflammatory cytokine.

Acupuncture Points ; Animals ; Antineoplastic Agents ; administration & dosage ; adverse effects ; Electroacupuncture ; Humans ; Interleukin-1beta ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasms ; drug therapy ; Peripheral Nervous System Diseases ; etiology ; genetics ; metabolism ; therapy ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

OBJECTIVETo explore the therapeutic basis of high dose intravenous immunoglobulin (IVIg) in treatment of peripheral neuropathy induced by Campylobacter jejuni lipopolysaccharide (CJ LPS).

METHOD(1) IVIg (400 mg/kg x d) was given to the rats on the different days respectively during the immunization with CJ LPS. Histological study of sciatic nerve was performed on the 35 th day after immunization. The titer of anti-CJ LPS antibody in sera of immunized rats was measured by ELISA; IgG deposition was detected by immunohistochemistry and expression of TNF-alpha mRNA in the pathological nerves by in situ hybridization histochemistry. (2) When PBMCs were stimulated by CJ LPS in vitro, IVIg was added into culture medium at the doses of 1, 2.5, 5, and 10 mg/ml, respectively. Pathological examination of sciatic nerve was performed on the 7th day after perineural injection of the supernatants. Expression of TNF-alpha mRNA in PBMCs stimulated by CJ LPS in medium was detected by in situ hybridization histochemistry after adding IVIg.

RESULTS(1) The rate of abnormal fibers appearance in IVIg group (1.0%) was much lower than that of the control group (15.0%) after immunization with CJ LPS, P < 0.01. The titer of antibody in control group was 9 times higher than that of IVIg group. There was no expression of immunoglobulin and TNF-alphamRNA in peripheral nerves in IVIg group, but high expression was found in control group in which no IVIg was injected. (2) The expression rates of TNF-alphamRNA on the PBMCs in IVIg group (1.0%) was much lower than that of control group (9.5%). (3) When the PBMCs of normal rats were stimulated by CJ LPS, the expression rates of TNF-alphamRNA in PBMCs of 5 mg/ml IVIg group (3.0%) or 10 mg/ml IVIg group (2.0%) were much lower than that of 1 mg/ml IVIg group (15.0%) or 2.5 mg/ml IVIg group (11.5%), P < 0.01. The rate of abnormal fibers appearance in 5 mg/ml IVIg group (9.8%) or 10 mg/ml IVIg group (8.5%) was much lower than that of 1 mg/ml IVIg group (50.0%), 2.5 mg/ml IVIg group (41.0%) or control group (50.8%) after the perineural injection with the supernatants, respectively, P < 0.01.

CONCLUSIONThe therapeutic effect of high dose IVIg might be associated with inhibition of the humoral and cellular immunity simultaneously in peripheral neuropathy induced by CJ LPS.

Animals ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulin G ; blood ; Immunoglobulins, Intravenous ; administration & dosage ; Immunohistochemistry ; In Situ Hybridization ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Peripheral Nervous System Diseases ; genetics ; immunology ; therapy ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; analysis ; genetics

OBJECTIVETo explore the important role of the terminal residues containing sialic acid (SA) in Campylobacter jejuni (CJ) lipopolysaccharide (LPS) as the critical antigen to induce nerve damage, and also to identify immunopathological evidence for the hypothesis of molecular mimicry and cross-immunity between CJ LPS and gangliosides.

METHODSA mutant of Pen O:19 CJ with neuB1 gene inactivated and LPS outer core terminal residues losing SA was to be constructed. PCR and RT-PCR were used to confirm the mutant. Capability of CJ LPS binding to cholera toxin B subunit (CTB) was tested. Guinea pigs were systematically immunized with LPS of the wild and the mutant strains, respectively. Titers of anti-LPS and anti-ganglioside GM(1) IgG antibodies in sera of immunized guinea pigs were detected by ELISA. Pathological study for sciatic nerves of both Guinea pigs either immunized systematically or perineural injection with their immunized serum was finished.

RESULTS(1) The mutant of CJ O:19 strain with inactivated neuB1 gene was successfully constructed and lost transcriptional activity of neuB1 gene in the mutant strain was confirmed by PCR and RT-PCR. SA was well demonstrated by both acidic ninhydrin reaction and periodate-resorcinol reaction in the LPS of wild strain but not in the mutant LPS; (2) Compared with the titers before immunization, the titers of anti-GM(1) IgG antibody increased in sera of guinea pigs immunized with LPS of the wild strain. However there were no detectable anti-GM(1) IgG antibody in sera of the animals immunized with mutant LPS and PBS. (3) The incidence of pathological fibers of sciatic nerves in wild CJ LPS group (17.3%) was significantly higher than the mutant CJ LPS group (chi(2) = 125, P < 0.01); the difference between the mutant CJ LPS group and control group was not statistically significant (chi(2) = 1.633, P > 0.05). (4) After perineural injection with immunized serum, the incidence of pathological fibers of sciatic nerves in wild strain group (67.8%) was also significantly higher than the incidence of mutant group (P < 0.01).

CONCLUSIONA mutant of CJ O:19 strain neuB1 gene inactivated and SA component of terminal structure of LPS lost was successfully constructed. And it no longer expressed SA component which is the normal terminal structure of LPS in wild strain. The capability of the wild strain to induce increased titers of anti-GM(1) antibody and immune-mediated nerve damage was simultaneously lost for the mutant strain. It could be a strong immunopathologic evidence to identify the molecular mimicry hypothesis between CJ LPS and ganglioside epitope in nerve on the pathogenesis of CJ related GBS. The terminal residues containing SA should be as the basic GM1-like structure in CJ LPS.

Animals ; Antibodies, Bacterial ; blood ; immunology ; Antigens, Bacterial ; genetics ; immunology ; Campylobacter jejuni ; genetics ; immunology ; G(M1) Ganglioside ; immunology ; Guinea Pigs ; Lipopolysaccharides ; chemistry ; immunology ; Molecular Mimicry ; Mutagenesis ; N-Acetylneuraminic Acid ; chemistry ; immunology ; Peripheral Nervous System Diseases ; immunology ; microbiology

Mutations and altered gene dosage of the peripheral myelin protein (PMP22) gene in chromosome 17p11.2-12 are the main causes for hereditary neuropathies, accounting for approximately 70% of all cases. Patients with duplication of the PMP22 develop Charcot-Marie-Tooth disease type 1A (CMT1A) and deletion of one PMP22 allele leads to hereditary neuropathy with liability to pressure palsy (HNPP). Twenty patients with CMT1A, 17 patients with HNPP, and 18 normal family members and 28 normal controls were studied by real-time quantitative PCR using SYBR Green I on the ABI 7700 Sequence Detection System. The copy number of the PMP22 gene was determined by the comparative threshold cycle method and the albumin was used as a reference gene. The PMP22 duplication ratio ranged from 1.45 to 2.06 and the PMP22 deletion ratio ranged from 0.42 to 0.64. The PMP22 ratio in normal controls, including normal family members, ranged from 0.85 to 1.26. No overlap was found between patients with CMT1A or patients with HNPP and normal controls. This method is fast, highly sensitive, specific, and reproducible in detecting PMP22 duplication and deletion in CMT1A and HNPP patients, respectively.
Charcot-Marie-Tooth Disease/*diagnosis/*genetics ; Chromosomes, Human, Pair 17 ; Family Health ; Female ; Fluorescent Dyes/*pharmacology ; Gene Deletion ; Gene Duplication ; Hereditary Motor and Sensory Neuropathies/*genetics ; Human ; Male ; Membrane Proteins/*biosynthesis ; Organic Chemicals/*pharmacology ; Paralysis/*genetics ; Peripheral Nervous System Diseases/*genetics ; Reverse Transcriptase Polymerase Chain Reaction