OBJECTIVE: To explore the clinical phenotypes and genetic characteristics of a child with Acid-labile subunit deficiency (ALS). METHODS: A male child diagnosed with ALS at Dongguan Maternal and Child Health Care Hospital in March 2021 was selected as the study subject. Clinical data of his family was collected. Peripheral blood samples were collected from the child and his parents. Following extraction of genomic DNA, whole-exome sequencing (WES) was carried out, and Sanger sequencing was used for family verification of candidate variants. Based on guidelines from the American College of Medical Genetics and Genomics (ACMG), the pathogenicity of the candidate variant was classified. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: 2020-6). RESULTS: The patient, a 5-year-and-7-month-old boy, presented with short stature and delayed bone age. Endocrine examinations showed decreased serum concentrations of insulin-like growth factor-1 (IGF-1) and IGF binding protein-3 (IGFBP3). WES revealed that he has harbored compound heterozygous variants of the IGFALS gene, namely c.741_742del, p.Y248Pfs83 and c.272del, p.P91Rfs31. Sanger sequencing verified that the variants were inherited from his father and mother, respectively. According to the ACMG guidelines, c.741_742del, p.Y248Pfs83 and c.272del, p.P91Rfs31 variants were classified as likely pathogenic (PVS1+PM2_supporting). Based on the pre-set literature search strategy, 11 research literature on ALS were retrieved, which involved a total of 33 families and 62 patients. Combined with the patient in this study, 31 IGFALS gene variants were identified among the 63 patients, which mainly consisted of missense variants (20 types), with variant sites concentrated in exon 2. The main clinical features were short stature in conjunct with delayed puberty, with a significant genotype-phenotype correlation. CONCLUSION The IGFALS gene variants NM_004970.2: c.741_742del, p.Y248Pfs83 and c.272del, p.P91Rfs31 may be the genetic etiology in this family. This study has expanded the variant spectrum of the IGFALS gene and provided valuable information for the diagnosis, genetic counseling and clinical treatment of the disease.
Humans ; Male ; Phenotype ; Child, Preschool ; Carrier Proteins/genetics* ; Glycoproteins/deficiency* ; Exome Sequencing ; Female ; Mutation ; Insulin-Like Growth Factor I/metabolism* ; Growth Disorders/genetics*

Neuroplastin 65 (Np65) is an immunoglobulin superfamily cell adhesion molecule involved in synaptic formation and plasticity. Our recent study showed that Np65-knockout (KO) mice exhibit abnormal cognition and emotional disorders. However, the underlying mechanisms remain unclear. In this study, we found 588 differentially-expressed genes in Np65-KO mice by microarray analysis. RT-PCR analysis also revealed the altered expression of genes associated with development and synaptic structure, such as Cdh1, Htr3a, and Kcnj9. In addition, the expression of Wnt-3, a Wnt protein involved in development, was decreased in Np65-KO mice as evidenced by western blotting. Surprisingly, MRI and DAPI staining showed a significant reduction in the lateral ventricular volume of Np65-KO mice. Together, these findings suggest that ablation of Np65 influences gene expression, which may contribute to abnormal brain development. These results provide clues to the mechanisms underlying the altered brain functions of Np65-deficient mice.
Affective Symptoms ; metabolism ; Animals ; Brain ; diagnostic imaging ; metabolism ; pathology ; Cognition Disorders ; metabolism ; Gene Expression ; Magnetic Resonance Imaging ; Membrane Glycoproteins ; deficiency ; genetics ; physiology ; Mice, Knockout ; Microarray Analysis ; Organ Size ; Real-Time Polymerase Chain Reaction ; Wnt3 Protein ; metabolism

Plants are known to be efficient hosts for the production of mammalian therapeutic proteins. However, plants produce complex N-glycans bearing β1,2-xylose and core α1,3-fucose residues, which are absent in mammals. The immunogenicity and allergenicity of plant-specific Nglycans is a key concern in mammalian therapy. In this study, we amplified the sequences of 2 plant-specific glycosyltransferases from Nicotiana tabacum L. cv Bright Yellow 2 (BY2), which is a well-established cell line widely used for the expression of therapeutic proteins. The expression of the endogenous xylosyltranferase (XylT) and fucosyltransferase (FucT) was downregulated by using RNA interference (RNAi) strategy. The xylosylated and core fucosylated N-glycans were significantly, but not completely, reduced in the glycoengineered lines. However, these RNAi-treated cell lines were stable and viable and did not exhibit any obvious phenotype. Therefore, this study may provide an effective and promising strategy to produce recombinant glycoproteins in BY2 cells with humanized N-glycoforms to avoid potential immunogenicity.
Amino Acid Sequence ; Blotting, Western ; Carbohydrate Sequence ; Cell Line ; Cloning, Molecular ; DNA, Complementary ; genetics ; Down-Regulation ; Epitopes ; genetics ; immunology ; Fucose ; metabolism ; Fucosyltransferases ; chemistry ; deficiency ; genetics ; immunology ; Glycoproteins ; chemistry ; genetics ; immunology ; Molecular Sequence Data ; Pentosyltransferases ; chemistry ; deficiency ; genetics ; immunology ; Polysaccharides ; chemistry ; immunology ; Protein Engineering ; methods ; RNA Interference ; Species Specificity ; Tobacco ; cytology ; genetics ; Xylose ; metabolism

Based on the NSP4 sequence of bovine rotavirus (BRV), the shRNA was designed and synthesized, and a shRNA recombinant lenti-virus vector RNAi-H1-89 was constructed. The recombinant RNAi-H1-89 Lenti-virus was packaged by transfecting the 293T cell with the recombinant vector RNAi-H1-89 and two helper plasmids using lipofectamine, and then used to infect MA104 cells. The MA104 cells were further infected with BRV strain G6 24h post-infection, with the LacZ shRNA recombinant lenti-virus as control. Thirty-six hours later, the CPE of the infected cells was observed under microscope, shRNA of NSP4 gene inhibited CPE in MA104 cell; the shRNA against NSP4 gene also inhibited NSP4 gene expression by RT-PCR, The virus titer in the cell culture supernatant was significant lower compared with the control group. The above results showed that RNAi-H1-89 against NSP4 gene could specifically silence NSP4 gene expression, and inhibit the proliferation of BRV.
Animals ; Base Sequence ; Cattle ; Cell Line ; DNA, Recombinant ; genetics ; Glycoproteins ; deficiency ; genetics ; Molecular Sequence Data ; Plasmids ; genetics ; RNA, Small Interfering ; genetics ; Rotavirus ; genetics ; physiology ; Toxins, Biological ; genetics ; Viral Load ; genetics ; Viral Nonstructural Proteins ; deficiency ; genetics ; Virus Replication ; genetics

BACKGROUNDOur previous research has suggested that platelet activating factor receptor was related to atherosclerosis. The present study investigated the effect of a platelet activating factor receptor antagonist-WEB2086 on angiogenesis in aortal plaque and ischemic hindlimb of apolipoprotein E-deficient mice.

METHODSEight-week-old apolipoprotein E-deficient mice were fed with a 0.15% cholesterol diet to develop advanced lesions. At age 32 weeks unilateral hindlimb ischemia was surgically induced and the mice were divided into two groups: with or without WEB2086 mixed with their drinking water (4.3 mg in 100 ml). At age 40 weeks blood was collected from the orbit for measurement of serum lipids and an enzyme linked immunosorbent assay was used to determine platelet activating factor and oxidized low density lipoprotein in the gastrocnemius and aorta. Whole-Mount CD31 stain and plaque-associated sprouting have been used to estimate angiogenesis in plaque from the aorta and laser Doppler perfusion imaging and immunohistochemical expression of von Willebrand factor have been used to estimate angiogenesis in ischemic hindlimb.

RESULTSThe lipid composition of serum was not different between the groups. However, the amount of platelet activating factor and oxidized low density lipoprotein detected in the aorta was significantly higher than that in the gastrocnemius of ischemic hindlimb. The ratio of lesion to aorta levels was significantly reduced by administration of WEB2086, (31.52 +/- 6.18)% vs (55.58 +/- 8.34)%, P < 0.01. The mean density of intimal capillaries in atherosclerotic plaque, (31.13 +/- 9.20)% vs (57.74 +/- 11.28)%, P < 0.01, and the mean number of sprouts per aorta were significantly reduced, 183.92 +/- 34.17 vs 392.54 +/- 76.79, P < 0.01, in the WEB2086 group. Blood flow (0.85 +/- 0.12 vs 0.45 +/- 0.06, P < 0.01) and capillary density of ischemic hindlimb (1.18 +/- 0.17 vs 0.53 +/- 0.09, P < 0.01) were markedly increased in apolipoprotein E-deficient mice treated with WEB2086 versus controls.

CONCLUSIONThe study provides evidence that WEB2086 can inhibit angiogenesis in atherosclerotic plaque but promote it in ischemic hindlimb.

Animals ; Apolipoproteins E ; deficiency ; Atherosclerosis ; physiopathology ; Azepines ; pharmacology ; Capillaries ; Cholesterol ; blood ; Hindlimb ; blood supply ; Ischemia ; physiopathology ; Lipoproteins, LDL ; blood ; physiology ; Male ; Mice ; Neovascularization, Physiologic ; drug effects ; Platelet Activating Factor ; analysis ; Platelet Membrane Glycoproteins ; antagonists & inhibitors ; Receptors, G-Protein-Coupled ; antagonists & inhibitors ; Triazoles ; pharmacology

OBJECTIVETo investigate the regulation pattern of the two compound prescriptions for Kidney tonifying, Yougui Yin and Bushen Yishou capsule, in down-regulating T-cell apoptosis gene expression in aged rats.

METHODSExpressions of T-cell apoptosis promoting and inhibiting genes, including Fas, FasL, Bcl-2, Bax, TNFR1 and TNFR2, as well as activity of cysteine proteinase in cascade connection, such as Caspase 8 and Caspase 3 were determined by TUNEL labeled flow cytometry and fluorescence real-time quantitative RT-PCR technique. The difference between old and young rats was compared, and the different regulation patterns of the two compound prescriptions for Kidney tonifying and their effects on Caspase activity were compared with those of compound prescription for blood circulation activating.

RESULTSThe two compound prescriptions for Kidney tonifying could effectively lower T-cell over-apoptosis in old rats, down-regulate FasL and TNFR1 gene transcription, and decrease the activity of Caspase 8 and Caspase 3, while the compound prescription for blood circulation activating showed insignificant effect on T-cell over-apoptosis.

CONCLUSIONKidney-deficiency is closely related to the T-cell over-apoptosis. The T-cell over-apoptosis in old rats could be effectively improved by the two compound prescription for Kidney tonifying through down-regulating the apoptosis promoting genes FasL and TN-FR1 transcription. That is the unique regulation pattern of Kidney tonifying principle to T-cell apoptosis related gene in old rats.

Aging ; drug effects ; genetics ; immunology ; Animals ; Apoptosis ; drug effects ; Blood Circulation ; physiology ; Caspase 3 ; Caspases ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; Female ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; Random Allocation ; Rats ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics ; T-Lymphocytes ; cytology ; Yang Deficiency ; immunology ; fas Receptor ; biosynthesis ; genetics

Glanzmann's thrombasthenia is a qualitative platelet disorder characterized by a deficiency in the platelet membrane glycoproteins IIb/IIIa. It belongs to a group of hereditary platelet disorders typified by normal platelet numbers and a prolonged bleeding time. The severity of bleeding does not correlate with the severity of the platelet glycoprotein IIb/IIIa abnormality. The present case report describes the periodontal treatment of a patient with Glanzmann's thrombasthenia. A 30-year-old female with a history of Glanzmann's thrombasthenia was referred for gingival bleeding on tooth brushing and discomforts in #38 area. The periodontal finding revealed a diagnosis of localized slight adult periodontitis. Root planing and extraction of #38 was performed under 12 pack of platelets transfusion and digital compression was done for hemostasis. The gingival bleeding ceased within a day in maxilla and 2 days later in mandible. 42 pack of platelets was administered for 3 days of post-treatment and for iron-deficiency anemia 3 pack of RBCs was transfused 2 days later. 1 week later the inflammation in gingiva disappeared and gingival stippling appeared. The clinical result we got was good and in such a medically compromised patient it is an ability to maintain a proper oral hygiene that is essential both for oral and systemic health.
Adult ; Anemia, Iron-Deficiency ; Bleeding Time ; Blood Platelet Disorders ; Blood Platelets ; Chronic Periodontitis ; Diagnosis ; Female ; Gingiva ; Glycoproteins ; Hemorrhage ; Hemostasis ; Humans ; Inflammation ; Mandible ; Maxilla ; Oral Hygiene ; Platelet Count ; Platelet Membrane Glycoproteins ; Root Planing ; Thrombasthenia* ; Tooth