BACKGROUND: Previous studies have confirmed that 6-hydroxydopamine is capable to increase the expression of divalent metal transporter-1 and reduce the expression of ferroportin-1 in the neurons and microglia, which may lead to iron deposition in the substantia nigra after Parkinson’s disease. However, it is unclear whether 6-hydroxydopamine can play diverse roles in astrocytes.
OBJECTIVE:To observe the effects of 6-hydroxydopamine on the expression of divalent metal transporter-1 and ferroportin-1 in rat C6 glioma cel lines.
METHODS:C6 glioma cell lines from rats were cultured in 10 μmol/L 6-hydroxydopamine for 24 hours. Then, protein expressions of divalent metal transporter-1 and ferroportiner-1 were measured by western blot method.
RESULTS AND CONCLUSION:The protein expressions of divalent metal transporter-1 and ferroportin-1 in C6 glioma cell lines were increased by 2.5 times (P < 0.01) and 1 time (P < 0.05), respectively, after treatment with 6-hydroxydopamine. These findings indicate that 6-hydroxydopamine can promote iron transport rate in astrocytes by increasing both divalent metal transporter-1 and ferroportin-1 expressions, and astrocytes has a different response to 6-hydroxydopamine from neurons and microglia.

AIM: To study the effects of insulin on the proliferation and function of osteoblasts and the relationship between insulin post-receptor change in osteoblasts and osteoblastic cell growth.METHODS: The effects of different levels of insulin on osteoblasts were assessed by MTT colorimetry.Osteocalcin in medium was measured by RIM.IGF-1 mRNA expression levels were determined by RT-PCR.The concentrations of free IGF-1 protein in serum-free medium were measured by ELISA.In addition,the protein level and phosphorylated protein of P~(44/42)MAPK were determined by Western blotting analysis.RESULTS: Insulin enhanced the proliferation of osteoblasts,depending on its dose and exposure time.Insulin at concentration of 10~(-7) mol/L showed the strongest effect,and the action attained the plateau phase beyond 96 h.The best concentration that stimulated synthesis of osteocalcin by insulin was 10~(-7) mol/L.When the insulin concentration beyond 10~(-7) mol/L,the osteocalcin concentration was decreased.Exposure time had no effect on insulin-stimulated synthesis of osteocalcin of osteoblastic cells.When the concentration of insulin reaches 10~(-6) mol/L,the IGF-1 mRNA expression stimulated by insulin was also decreased.The concentrations of free IGF-1 protein in insulin-stimulated groups were all higher than that in control group(P0.05).Insulin acute stimulation rapidly induced the activity of tyrosine phosphorylation of P~(44/42)MAPK.The degree of tyrosine phosphorylation of P~(44/42)MAPK was increased step by step along with the increasing doses of insulin from 0 to 10~(-7) mol/L(P

Objective To analyze the serotypes of enteroviruses(EVs) isolated from patients with influenza-like illness in Beijing in 2017. Methods Oropharyngeal swab specimens were collected from pa-tients with influenza-like illness in eight districts of Beijing from July 2017 to October 2017. EVs were detec-ted by real-time PCR. Specific primers were synthesized and used to amplify the VP1 fragments of EVs. PCR products were sequenced and the results were compared with the reference sequences by using Basic Lo-cal Alignment Search Tool(BLAST) to identify the serotypes of isolated EVs. Results A total of 666 spec-imens were collected and 91 (13.66%) were positive for EVs. VP1 sequences of 66 EVs were successfully amplified and BLAST analysis revealed that these strains belonged to 14 serotypes,including seven serotypes of EV-A species,six serotypes of EV-B species and one serotype of Rhinovirus species. The predominant se-rotypes were CVA2 and CVA6. Eight out of 14 CVA6 strains that were collected in Shunyi District shared high homology. All seven CVB5 strains were collected in Shijingshan District and grouped into one cluster. Conclusion EVs causing influenza-like illness in Beijing in 2017 belonged to 14 serotypes and CVA2 and CVA6 were the predominant serotypes.

Objective:To summarize the prenatal ultrasound features of fetal cleidocranial dysplasia (CCD) and provide references for clinical consultation.Methods:A retrospective analysis was conducted on the prenatal ultrasound features, genetic testing results, and prognosis of a CCD fetus diagnosed at Qingdao Women and Children's Hospital in June 2023. Relevant literature on CCD was retrieved from the CNKI, Yiigle, Wanfang, and PubMed databases including cases confirmed by genetic testing or postnatal clinical phenotype and imaging with relatively complete prenatal ultrasound information. The prenatal ultrasound features of CCD fetuses were summarized using descriptive statistical analysis.Results:(1) In this case, prenatal ultrasound at 25 weeks of gestation indicated widened cranial sutures with clear near-field intracranial structures, absence of the nasal bone, shortened and rigid bilateral clavicles, and mildly shortened bilateral femurs and humeri. Chromosomal karyotyping and chromosomal microarray analysis showed no abnormalities, but whole exome sequencing detected a RUNX2 gene mutation, leading to a diagnosis of CCD combined with the ultrasound phenotype. At 36 weeks of gestation, the mother experienced premature rupture of membranes and delivered a male infant vaginally. Bedside X-rays indicated bilateral wet lungs and bilateral clavicular dysplasia in the newborn. Telephone follow-up at 9 months showed no abnormalities in growth and development. (2) Literature review: Thirteen cases from 13 articles were included, along with this case, totaling 14 CCD fetuses. The main ultrasound phenotypes of CCD fetuses were clavicular dysplasia (12/14), incomplete cranial ossification (10/14), absence of the nasal bone (8/14), and shortening of the femur (12/14). Other ultrasound phenotypes included scapular dysplasia, short ribs, and increased interocular distance. Conclusion:Clavicular dysplasia and incomplete cranial ossification are the most specific signs of fetal CCD, while absence of the nasal bone and mild femoral shortening are secondary features of CCD.

Objective:To improve clinicians′ understanding of developmental and epileptic encephalopathy (DEE) caused by PPP3CA gene mutation. Methods:Clinical data of a patient with DEE diagnosed in the First Department of Neurology, Hebei Children′s Hospital in September 2018 were collected. The whole-exome sequencing of the proband′s family was performed, and the characteristics of gene mutation were analyzed. Literature review was carried out based on the reported cases related to PPP3CA gene. Results:The proband, a 3 months and 20 days old girl, was admitted to the hospital with a history of paroxysmal confusion with extremities shaking for 2 days. The clinical manifestations included frequent epilepsy seizures and hypoevolutism. Brain magnetic resonance imaging showed that the bilateral frontotemporal extracerebral space was slightly wider. The video electroencephalography showed hyperarrhythmia and a cluster of spastic seizures. Whole exome sequencing of the family revealed that the proband had a heterozygous de novo frameshift truncating mutation in the PPP3CA gene: c.1255-1256delAG (p.Ser419Cysfs*31). From the establishment of the database to May 2022, 8 foreign literatures and 1 Chinese literature were retrieved, and a total of 21 children with PPP3CA gene mutation were reported, with clinical developmental delay, cognitive dysfunction and abnormal electroencephalography activity. Conclusions:The frameshift truncating mutation of the PPP3CA gene (c.1255-1256delAG) is the hereditary etiology of this patient. For cases of frequent seizures with poor efficacy of antiepileptic drugs, and developmental delay, genetic testing should be performed to confirm diagnosis and treatment.

ObjectiveTo investigate the material basis and mechanism of Arnebia euchroma against hepatocarcinoma by network pharmacology， and to verify the potential targets of A. euchroma against hepatocarcinoma by molecular docking and experiments. MethodThe main active ingredients of A. euchroma were collected by retrieving the literature through China National Knowledge Network （CNKI） and Traditional Chinese Medicine System Pharmacology Database and Analysis Platform（TCMSP）. The active ingredients were screened out by FAFDrug4 platform according to the pharmacokinetics （ADME） properties of the drugs. The screening compounds and liver cancer targets were collected by using several databases and analyzed by drawing Venn diagrams. The protein-protein interaction （PPI） network was constructed by Cytoscape and STRING database. DAVID database was used to perform Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis of key targets. Autodock was used to perform molecular docking of targets on core pathways. Cell counting kit-8 （CCK-8） experiment was carried out to validate the activities of five naphthoquinones. Based on the predicted results in the H22 tumor-bearing mouse model， the key targets of isovalerylshikonin against hepatocarcinoma were verified by hematoxylin-eosin （HE） staining， enzyme-linked immunosorbent assay （ELISA）， and Western blot assay. ResultFifty-five active ingredients and 34 targets of active ingredients against hepatocarcinoma were screened out. The active molecules with high degree values in the “drug-active ingredient-target-disease” network were mainly naphthoquinones. PPI network obtained several core targets of A. euchroma against hepatocarcinoma. Twenty-two pathways were screened out by KEGG analysis， mainly involving phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）， vascular endothelial growth factor （VEGF）， tumor necrosis factor （TNF）， and other signaling pathways. The results of molecular docking showed that the five naphthoquinones had a good affinity with the targets of the PI3K/Akt signaling pathway. The results of CCK-8 and animal experiments showed that the lipid-soluble component isovalerylshikonin had good anti-cancer potential， and the high-dose group reduced the serum levels of VEGF and alpha fetoprotein （AFP） levels and elevated interferon-γ （IFN-γ） level （P<0.05， P<0.01）. The high-dose group also down-regulated phosphorylate（p）-Akt （Ser473） and B-cell lymphoma （Bcl）-2 protein expressions and up-regulated Bcl-2-antagonist of cell death （Bad） protein expression （P<0.01）. ConclusionA. euchroma can inhibit hepatocarcinoma cell proliferation and tumor angiogenesis and induce cancer cell apoptosis through the PI3K/Akt signaling pathway， which provides ideas and clues for the subsequent in-depth investigation of its specific mechanism.

Amiloride ; analogs & derivatives ; chemistry ; metabolism ; Binding Sites ; Genotype ; Hepacivirus ; genetics ; metabolism ; Humans ; Magnetic Resonance Spectroscopy ; Molecular Dynamics Simulation ; Protein Structure, Tertiary ; Viral Proteins ; antagonists & inhibitors ; metabolism