The molecular mechanism of verbascoside(OC1) in promoting acetylcholine(ACh) release in the pathogenesis of Alzheimer's disease(AD) was studied. Adrenal pheochromocytoma cells(PC12) of rats induced by β-amyloid protein(1-42)(Aβ_(1-42)) were used as AD models in vitro and were divided into control group, model group(Aβ_(1-42) 10 μmol·L~(-1)), OC1 treatment group(2 and 10 μg·mL~(-1)). The effect of OC1 on phosphorylated proteins in AD models was analyzed by whole protein phosphorylation quantitative omics, and the selectivity of OC1 for calcium channel subtypes was virtually screened in combination with computer-aided drug design. The fluorescence probe Fluo-3/AM was used to detect Ca~(2+) concentration in cells. Western blot analysis was performed to detect the effects of OC1 on the expression of phosphorylated calmodulin-dependent protein kinase Ⅱ(p-CaMKⅡ, Thr286) and synaptic vesicle-related proteins, and UPLC/Q Exactive MS was used to detect the effects of OC1 on ACh release in AD models. The effects of OC1 on acetylcholine esterase(AChE) activity in AD models were detected. The results showed that the differentially modified proteins in the model group and the OC1 treatment group were related to calcium channel activation at three levels: GO classification, KEGG pathway, and protein domain. The results of molecular docking revealed the dominant role of L-type calcium channels. Fluo-3/AM fluorescence intensity decreased under the presence of Ca~(2+) chelating agent ethylene glycol tetraacetic acid(EGTA), L-type calcium channel blocker verapamil, and N-type calcium channel blocker conotoxin, and the effect of verapamil was stronger than that of conotoxin. This confirmed that OC1 promoted extracellular Ca~(2+) influx mainly through its interaction with L-type calcium channel protein. In addition, proteomic analysis and Western blot results showed that the expression of p-CaMKⅡ and downstream vesicle-related proteins was up-regulated after OC1 treatment, indicating that OC1 acted on vesicle-related proteins by activating CaMKⅡ and participated in synaptic remodeling and transmitter release, thus affecting learning and memory. OC1 also decreased the activity of AChE and prolonged the action time of ACh in synaptic gaps.
Animals ; Rats ; Glucosides/administration & dosage* ; Acetylcholine/metabolism* ; Alzheimer Disease/genetics* ; PC12 Cells ; Phenols/chemistry* ; Neurotransmitter Agents/metabolism* ; Drugs, Chinese Herbal ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/genetics* ; Humans ; Phosphorylation/drug effects* ; Calcium/metabolism* ; Polyphenols

OBJECTIVE: To evaluate the effectiveness of minimally invasive reduction and ulna bicortical angulation intramedullary fixation in the management of Bado typeⅠMonteggia fracture in pediatric patients. METHODS: Between August 2018 and August 2020, the clinical data of 15 pediatric patients diagnosed with Bado typeⅠfresh Monteggia fracture were retrospectively analyzed. There were 11 males and 4 females. The left side was implicated in 5 cases, while the right side was involved in 10 cases. The age ranged from 4 to 12 years old. There were a total of 10 cases of upper ulna fractures and 5 cases of middle ulna fractures. Among these, the ulnar fracture line presented as transverse or short oblique in 9 cases, and long oblique or spiral in 6 cases. The pediatric patients underwent treatment utilizing minimally invasive reduction and ulna bicortical angulation intramedullary fixation. The effectiveness of the treatment was assessed based on the Mayo elbow scoring system. RESULTS: The follow-up period for all 15 cases ranged from 6 to 24 months. Among them, the occurrence of needle tail irritation was observed in 3 cases, all of which exhibited satisfactory healing following routine disinfection and dressing change. The other children had no re dislocation of the radial capitulum, no epiphyseal injury, no osteofascial compartment syndrome, elbow instability and other complications. The degree of limitation of daily life, range of motion of joints, pain level and appearance of limbs were recorded at the last follow-up. According to Mayo elbow scoring system, 12 cases were excellent and 3 cases were good. CONCLUSION The minimally invasive reduction and ulna bicortical angulation intramedullary fixation technique is a simple and effective treatment for Bado typeⅠfresh Monteggia fractures in children.
Humans ; Male ; Female ; Child ; Monteggia's Fracture/surgery* ; Child, Preschool ; Minimally Invasive Surgical Procedures/methods* ; Fracture Fixation, Intramedullary/methods* ; Retrospective Studies

OBJECTIVE: To investigate the clinical efficacy and safety of ultrasound-guided closed reduction and internal fixation using Kirschner wire for the treatment of olecranon fractures of the ulna in children. METHODS: Between January 2019 and January 2021, 13 children with olecranon fracture were treated with ultrasound-guided closed reduction and percutaneous Kirschner wire internal fixation, including 10 males and 3 females. The age ranged from 3 to 14 years old. Children with ulnar olecranon fractures were evaluated using the Gicquel scoring system. The clinical evaluation encompassed postoperative pain, functional status, and range of motion, with a maximum score of 15 points. The radiological assessment contributed an additional 4 points. A cumulative score of more than 18 scores was classified as excellent, more than 17 scores as good, more than16 scores as fair, and less than 16 scores as poor. Clinical assessment:A score of 14 indicates excellent performance, a score of 13 reflects good performance, a score of 12 denotes fair performance, and a score of less than 11 signifies poor performance. RESULTS: A total of 13 patients were followed up, with a duration ranging from 6 to 12 months. According to the Gicquel scoring criteria, the comprehensive evaluation of clinical and radiographic findings yielded 10 excellent and 3 good outcomes. Evaluation based solely on clinical findings resulted in 13 excellent outcomes. CONCLUSION Ultrasound-guided percutaneous cross Kirschner wire fixation for children's olecranon fracture has the advantages of less trauma, rapid recovery, less fluoroscopy, and good recovery of elbow function. The clinical effect is satisfactory.
Humans ; Child ; Male ; Female ; Fracture Fixation, Internal/instrumentation* ; Ulna Fractures/physiopathology* ; Bone Wires ; Child, Preschool ; Adolescent ; Olecranon Process/surgery* ; Ultrasonography ; Closed Fracture Reduction/methods* ; Olecranon Fracture

The patient, an 11-year-old girl, was admitted with recurrent fever for 20 days, worsening with abdominal distension for 7 days. Upon admission, she presented with recurrent fever, lymphadenopathy, hepatosplenomegaly, polyserositis, and multiple organ dysfunction. Lymph node pathology and clinical manifestations confirmed the diagnosis of idiopathic multicentric Castleman disease-TAFRO syndrome. Treatment with siltuximab combined with glucocorticoids was initiated, followed by maintenance therapy with tocilizumab. The patient is currently in complete clinical remission. Therefore, once a child is diagnosed with idiopathic multicentric Castleman disease -TAFRO syndrome, early use of siltuximab should be considered for rapid disease control, followed by tocilizumab for maintenance therapy.
Humans ; Castleman Disease/drug therapy* ; Child ; Antibodies, Monoclonal, Humanized/administration & dosage* ; Female ; Antibodies, Monoclonal/administration & dosage*

OBJECTIVES: To investigate the molecular epidemiology of children with phenylketonuria (PKU) in Gansu, China, providing foundational data for intervention strategies. METHODS: A retrospective analysis was conducted on 1 159 PKU families who attended Gansu Provincial Maternity and Child Care Hospital from January 2012 to December 2024. Sanger sequencing, multiplex ligation-dependent probe amplification, whole exome sequencing, and deep intronic variant analysis were used to analyze the PAH gene. RESULTS: For the 1 159 children with PKU, 2 295 variants were identified in 2 318 alleles, resulting in a detection rate of 99.01%. The detection rates were 100% (914/914) in 457 classic PKU families, 99.45% (907/912) in 456 mild PKU families, and 96.34% (474/492) in 246 mild hyperphenylalaninemia families. The 2 295 variants detected comprised 208 distinct mutation types, among which c.728G>A (14.95%, 343/2 295) had the highest frequency, followed by c.611A>G (4.88%, 112/2 295) and c.721C>T (4.79%, 110/2 295). The cumulative frequency of the top 23 hotspot variants reached 70.28% (1 613/2 295), and most variant alleles were detected in exon 7 (29.19%, 670/2 295). CONCLUSIONS Deep intronic variant analysis of the PAH gene can improve the genetic diagnostic rate of PKU. The development of targeted detection kits for PAH hotspot variants may enable precision screening programs and enhance preventive strategies for PKU.
Humans ; Phenylketonurias/epidemiology* ; Female ; Male ; Retrospective Studies ; Phenylalanine Hydroxylase/genetics* ; Mutation ; Child, Preschool ; China/epidemiology* ; Child ; Infant

OBJECTIVE: To investigate the relationship among seminal oxidation-reduction potential (nORP), sperm DNA fragmentation (DFI) and semen parameters in patients with varicocele. METHODS: Clinical data of 522 patients treated in the reproductive andrology clinic of the Northern Theater General Hospital from November 2023 to December 2023 were retrospectively analyzed, including 435 men of childbearing age and 87 men of infertile age. The patients were divided into the varicocele group (n=116) and non-varicocele group (n=406) according to clinical diagnosis. The differences of seminal plasma nORP, DFI, sperm high DNA stain ability (HDS) and semen parameters were analyzed between the two groups. The relationship among general clinical data, seminal plasma nORP, semen parameters, DFI and HDS in patients with varicocele were further analyzed. According to the severity of varicocele, the patients were divided into three groups, including mild, moderate and severe. And the differences of seminal plasma nORP and semen parameters, DFI and HDS among all groups were analyzed. The differences of seminal plasma nORP, semen parameters, DFI and HDS were compared between the varicocele and non-varicocele groups. RESULTS: The total sperm count, sperm concentration, progressive motility sperm percentage (PR%) and normal sperm morphology rate (NSMR) in patients with varicocele were significantly lower than those in control group (P<0.05). And seminal plasma nORP, DFI and HDS in patients with varicocele were significantly higher than those in control group (P<0.05). Seminal plasma nORP in patients with varicocele was significantly negatively correlated with total sperm, sperm concentration and NSMR (P<0.05), and significantly positively correlated with DFI and HDS (P<0.05). There were significant differences in nORP, total sperm count, sperm concentration, PR%, DFI and HDS among mild, moderate and severe varicocele groups (P<0.05). Seminal plasma nORP, sperm concentration, PR% and DFI in severe group were significantly lower than those in mild and moderate groups(P<0.05). Sperm count and HDS in severe group were significantly lower than those in mild group (P<0.05). In infertile patients, seminal plasma nORP, DFI and HDS in varicocele group were significantly higher than those in control group (P<0.05). And PR% in varicocele group was significantly lower than that in control group (P<0.05). CONCLUSIONS Seminal plasma nORP in patients with varicocele may be an important marker of oxidative stress affecting DFI and semen parameters.
Humans ; Male ; Varicocele/metabolism* ; Semen/metabolism* ; Spermatozoa ; Sperm Count ; Infertility, Male ; Retrospective Studies ; DNA Fragmentation ; Oxidation-Reduction ; Semen Analysis ; Adult ; Sperm Motility

Objective:To explore the genetic basis for a Chinese pedigree affected with co-morbid Ornithine carbamoyl transferase deficiency (OTCD) and MECP2 duplication syndrome.Methods:A proband who was admitted to the Neonatal Intensive Care Unit of Gansu Provincial Maternal and Child Health Care Hospital on December 19, 2017 was selected as the study subject. High-throughput sequencing and multiplex ligation-dependent probe amplification (MLPA) were carried out for her pedigree, and short tandem repeat-based linkage analysis and chromosome copy number variation sequencing (CNV-seq) were used for the prenatal diagnosis.Results:The proband, a 3-day-old female, was found to harbor heterozygous deletion of exons 7-9 of the OTC gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as likely pathogenic (PVS1+ PM2_Supporting+ PP4). The proband was diagnosed with OTCD, which was in keeping with her acute encephalopathy and metabolic abnormalities (manifesting as hyperammonemia, decreased blood citrulline, and increased urine orotic acid). Prenatal diagnosis was carried out for the subsequent pregnancy. The fetus did not harbor the exons 7-9 deletion of the OTC gene, but was found to carry a duplication in Xq28 region (which encompassed the whole region of MECP2 duplication syndrome) and was positive for the SRY sequence. The same duplication was also found in the proband and her mother. Considering the possible existence of X-chromosome inactivation, the proband was diagnosed with two X-linked recessive disorders including OTCD and MECP2 duplication syndrome, and the fetus was determined as a male affected with the MECP2 duplication syndrome. Conclusion:Discoveries of the pathogenic variants underlying the OTCD and MECP2 duplication syndrome have enabled clinical intervention, treatment, genetic counseling and prenatal diagnosis for this pedigree.

Objective:To analyze the clinical phenotype and genotypes of two children with Carnitine-acylcarnitine translocase deficiency (CACTD).Methods:Two children diagnosed with CACTD at the Gansu Provincial Maternal and Child Health Care Hospital respectively on January 3 and November 19, 2018 were selected as the study subjects. Trio-whole exome sequencing (trio-WES) was carried out, and candidate variants were validated through Sanger sequencing and pathogenicity analysis.Results:Both children were males and had manifested mainly with hypoglycemia. Trio-WES and Sanger sequencing showed that child 1 had harbored compound heterozygous variants of the SLC25A20 gene, namely c. 49G>C (p.Gly17Arg) and c. 106-2A>G, which were inherited from his father and mother, respectively. Child 2 had harbored homozygous c. 199-10T>G variants of the SLC25A20 gene, which were inherited from both of his parents. Among these, the c. 106-2A>G and c. 49G>C variants were unreported previously. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 49G>C (p.Gly17Arg), c. 106-2A>G, and c. 199-10T>G variants were classified as likely pathogenic (PM2_supporting+ PP3+ PM3_strong+ PP4), pathogenic (PVS1+ PM2_supporting+ PM5+ PP3), and pathogenic (PVS1+ PM2_supporting+ PP3+ PP5), respectively. Conclusion:Combined with their clinical phenotype and genetic analysis, both children were diagnosed with CACTD. Above finding has provided a basis for their treatment as well as genetic counseling and prenatal diagnosis for their families.

Objective To investigate the effect of vecuronium bromide on the malignant biological behavior of gastric cancer cells and its molecular mechanism.Methods Gastric cancer cells HGC-27 were divided into control group,experimental-L,-M,-H groups,si-NC group,si-FGD5-AS1 group,pcDNA-FGD5-AS1 group,pcDNA group,experimental-H+pcDNA group,experimental-H+pcDNA-FGD5-AS1 group.The control group was cultured conventionally;experimental-L,-M,-H groups were treated with 5,10 and 20μmol·mL-1 vecuronium bromide,respectively;si-FGD5-AS1 group and the si-NC group were transfected with lncRNA FGD5-AS1 interference expression vector and negative control plasmid,respectively;pcDNA-FGD5-AS1 group and pcDNA group were transfected with lncRNA FGD5-AS1 overexpression vector and negative control plasmid,respectively;lncRNA FGD5-AS1 overexpression vector and negative control plasmid were transfected into HGC-27 cells in the experimental-H+pcDNA-FGD5-AS1 group and experimental-H+pcDNA group,and then treated with 20 μmuol·mL-1 vecuronium bromide.Methyl thiazolyl tetrazolium(MTT),flow cytometry and real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)were applied to dectect cell viability,apoptosis and lncRNA FGD5-AS1 expression.Results The cell activity of control group,experimental-L,-M,-H groups,si-NC group,si-FGD5-AS1 group,pcDNA group,PCDNA-FGD5-AS1 group,experimental-H+pcDNA group and experimental-H+PCDNA-FGD5-AS1 group were 1.31±0.07,0.58±0.03,1.31±0.06,0.51±0.03,1.29±0.08,1.68±0.15,0.59±0.03 and 1.16±0.06;the apoptosis rates were(6.49±0.44)％,(23.52±0.98)％,(6.42±0.44)％,(26.75±0.97)％,(6.72±0.38)％,(2.56±0.19)％,(23.56±1.04)％and(11.65±0.47)％;the expression levels of lncRNA FGD5-AS1 were 1.00±0.05,0.37±0.02,0.99±0.05,0.21±0.02,1.00±0.03,2.98±0.12,0.38±0.02 and 0.87±0.05,respectively.The above indexes were compared with the control group and experimental-H group,those in the si-FGD5-AS1 group were compared with the si-NC group,those in the pcDNA-FGD5-AS1 group were compared with the pcDNA group,and those in the experimental-H+pcDNA-FGD5-AS1 group were compared with the experimental-H+pcDNA group,the differences were statistically significant(all P＜0.05).Conclusion Vecuronium bromide may inhibit the proliferation of HGC-27 cells and promote cell apoptosis by down-regulating lncRNA FGD5-AS1.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.