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.

Objective To observe the prenatal ultrasonic manifestations and outcomes of fetal spinal cord terminal central duct dilation.Methods Data of 8 fetuses with spinal cord terminal central duct dilation detected with prenatal ultrasound were retrospectively analyzed,and the ultrasonic manifestations and outcomes were observed.Results Prenatal ultrasound showed varying degrees of spinal cord terminal central duct dilation in all 8 fetuses.The cystic dilation area disappeared spontaneously in 4 fetuses,gradually increased in 1 fetus,continued to exist without significant changes in 1 fetus,the echo at the end of the expansion zone gradually changed from good sound transmission to uneven in 1 fetus,and the cystic expansion structure disappeared and the central duct end enlarged as a whole in third trimester in 1 fetus.Among 8 newborns,no significant abnormality was detected with physical examination in 6 cases,including 1 case of 1-month-old MRI showed fatty filum terminale and 1 case of 3-month-old MRI showed spinal canal cyst and fatty filum terminale,the other 2 cases were diagnosed as tethered cord syndrome and filum terminale fibrolipoma with physical examination and MRI.Conclusion Prenatal ultrasonic manifestations of fetal spinal cord terminal central duct dilation included cystic hypoechoic area of the spinal cord cone and adjacent structures,with clear boundaries and good sound transmission.However,the outcomes of fetal spinal cord terminal central duct dilation were somewhat different.

BACKGROUND:Synthetic polymers,such as polypropylene and polyester,used for the treatment of abdominal wall defects not only lack biodegradability and bioactivity but also fail to meet the demands of complex and irregular wounds.Therefore,finding bioactive materials with low immunogenicity and good histocompatibility has become a hot spot in the repair of abdominal wall defects. OBJECTIVE:To prepare methacryloyl modified dermal extracellular matrix hydrogel and explore its potential application in abdominal wall defect. METHODS:(1)The porcine dermis was acellular with 0.25%trypsin and 1%Triton X-100 in turn to obtain the dermal extracellular matrix.After pepsin digestion and methacrylic anhydride modification,the methacrylated dermal extracellular matrix hydrogel was formed by photocrosslinking.The microscopic morphology of the hydrogel was observed by scanning electron microscope,and its rheological properties,swelling properties and other physical and chemical properties were tested.(2)Mice fibroblasts(L929)were inoculated into methacrylated dermal extracellular matrix hydrogel to detect the cell compatibility.(3)Totally 12 SD rats were randomly divided into two groups(n=6)to create abdominal wall defect model with peritoneum preserved.The defect site of the polypropylene group was filled with polypropylene material,and the hydrogel group was filled with methacrylated dermal extracellular matrix hydrogel.The wound skin of both groups was covered with polypropylene material.The wound healing was observed and histological analysis was carried out. RESULTS AND CONCLUSION:(1)Enzymatic hydrolysis had a good decellularization effect on porcine dermis after decellularization,and the original glycosaminoglycans and collagen were well retained.Scanning electron microscope observation revealed that the dermal extracellular matrix hydrogel presented loose and porous structure.The aperture was between 70 and 120 μm.The swelling ratio was(16.88±3.24)%and the water absorption was(94.24±1.11)%.The rheological property test showed that the methacrylated dermal extracellular matrix hydrogel was stable and had shear thinning characteristics,with injectability.(2)CCK-8 assay and live/dead staining showed that methacrylated dermal extracellular matrix hydrogel had good cell compatibility.(3)The results of animal experiments showed that the skin wound healing rate of the experimental group was higher than that of the control group at 7,10,and 14 days after operation(P<0.05).Hematoxylin-eosin and Masson staining of skin and muscle tissue exhibited that compared with the polypropylene group,the skin wound epithelialization,hair follicle formation,collagen fiber arrangement,and neovascularization were better in the hydrogel group 14 days after surgery.The skin wound new tissue structure was similar to the normal tissue at 28 days after surgery,and scar hyperplasia was less.A small amount of muscle regeneration was observed on day 28 after operation.(4)The results show that the methacrylated dermal extracellular matrix hydrogel can promote wound skin healing and muscle tissue regeneration in rats with abdominal wall defect.