Objective To develop a liquid chromatography-mass spectrometry (LC-MS) method for the determination of S-allyl-L-cystein in extraction of garlic. Methods A LC-MS method was established on a ZORBAX Eclipse XDB-C8(4.6 mm×250 mm, 5 μm)column with the mobile phase consisting of 1‰ formic acid water-methanol (95:5), flow rate of 0.8 mL/min and detection with post-column splitting. The split ratio was 2:1, column temperature was set at 25 ℃. The mass spectrometer equipped with ESI and the ion source was operated in negative mode. The dry gas flow was 10.0 L/min, the nebulizer pressure was 30 psig, and the vaporizer temperature was 350 ℃. SIM detector, S-allyl-L-cystein m/z 160 and S-allyl-L-cystein sulfoxide m/z 176. Results The calibration curves showed good linearity in the range of 0.062 5-2 μg/mL of S-allyl-L-cystein. The detection limit was 0.01 μg/mL. The within-day RSD was 4.11% and the day-to-day RSD was 4.49%.The average recovery for S-allyl-L-cystein was 101.63%. Conclusions The method is simple and accurate. It is adapted to determine and analyze S-allyl-L-cystein in extraction of garlic and the average content of S-allyl-L-cystein is 0.514 μg/mg.

To investigate the protection effect of DNA vaccine in mammalian and avian systems, the DNA vaccine was inoculated in both BALB/c mice and SPF chickens immunized with DNA vaccines encoding hemagglutinin (HA) from A/Goose/GuangDong/1/96 (H5N1) virus. The mice and chickens were immunized twice, 3 weeks apart, by electroporation into muscles or intramuscular injection. Two weeks after the second immunization, the mice and chickens were challenged with a lethal dose of homologous virus. The mice and chickens immunized by electroporation obtained completely protection against the virus, and could effectively inhibited viruses to replicating in mouse lung and chicken cloaca. At the same time, these protections were companied by high levels specific antibody to H5N1 AIV, while the blank plasmid controls experience 100 percent mortality following challenge. Furthermore, in the experiment of mice by eletroporation,stronger obviously CTL activity were observed after challenge. Thus, the cellular immune responses of the mice immunized by electroporation were exhibited. These results strongly demonstrate that HA DNA vaccines provide effective protection against influenza virus infection in mammalian and avian, and suggest that electroporation is one of the efficient gene delivery systems for the transfer of influenza DNA vaccine in both humoral immunity and cellular immunity.

Scoliosis is a common deformity in neuromuscular disease, which usually has the characteristics of early onset age, severe degree of deformity, and rapid progression.Neuromuscular scoliosis often leads to serious damages to the quality of life, and results in the loss of walking, standing and sitting, and cardiopulmonary insufficiency.Surgical treatment can improve the quality of life for children with neuromuscular scoliosis, but surgical treatment is still challenging due to the complex surgery and many complicated diseases.The complications are much higher than those of idiopathic scoliosis.A multidisciplinary team is necessary in the surgical treatment of neuromuscular scoliosis to promptly and effectively reduce the complications.

Objective:To summarize the characteristics of autosomal recessive axonal neuropathy with neuromyotonia (ARAN-NM) caused by HINT1 gene mutation. Methods:Retrospective case summary.Clinical data of 2 Tibetan siblings diagnosed with ARAN-NM in the Department of Pediatrics of Peking University First Hospital in August 2023 were retrospectively analyzed.A review of literature reporting relevant Chinese patients was conducted.Results:The proband and her elder brother were aged 13 and 19, respectively.Both developed abnormal gait at the age of 11, followed by varus, claudication, and weak thumb strength.The proband also had neuromyotonia.Physical examinations showed that the proband and her elder brother had decreased muscle strength of the extremities, mainly in the thumbs and distal ends of lower limbs.The distal muscles of the proband′s lower extremities and the muscles of both hands of the proband′s elder brother were atrophied.Both feet showed talipes equinovarus in the proband and her elder brother.The proband′s electromyography (EMG) showed peripheral nerve injuries (motor and sensory axonal involvement, especially in distal ends) and myotonic potentials.The trio-whole exon sequencing detected homozygous pathogenic variation in HINT1 gene in both the proband and her elder brother, who were diagnosed as ARAN-NM based on c. 169A>G (p.K57E). After the Carbamazepine treatment, the proband′s neuromyotonia, numbness and weakness were relieved.Both the proband and her elder brother underwent orthopaedic surgery and rehabilitation.Their foot deformities and gait were significantly improved.Two Chinese literatures (2 patients) and four English literatures (8 patients) were retrieved.Including the proband and her elder brother in this study, there were 12 ARAN-NM patients, 10 of whom had clinical data.The ages of onset and diagnosis were 2-16 (1 case unknown) and 13-33 years old, respectively.Myasthenia was present in 9 patients, especially in distal ends.Eight patients were complicated with neuromyotonia, nine patients with muscle atrophy, seven patients with foot deformity, and two patients with sensory disturbance.Creatine kinase(CK) was elevated in all 9 patients tested or CK.EMG showed neurogenic injuries in all patients and neuromyotonia discharge in six patients.Three patients were treated with Carbamazepine, and some symptoms were relieved.Missense/nonsense mutations were found in the 12 patients, and the high-frequency variation was c. 112T>C (p.C38R). Conclusions:ARAN-NM is a rare autosomal recessive neuromuscular disease caused by HINT1 gene mutation.There is no ethnic difference in clinical manifestations, mainly distal limb weakness with neuromyotonia.Carbamazepine can alleviate some symptoms, and orthopaedic surgery can improve foot deformity and gait.