OBJECTIVE: To explore the clinical characteristics and genetic cause of a child with gastrointestinal hemorrhage and Cerebroretinal microangiopathy with calcifications and cysts (CRMCC) and to review the literature. METHODS: Clinical data of a child with gastrointestinal hemorrhage with CRMCC admitted to the Hepatology Department of Hunan Children's Hospital in September 2019 were collected, and peripheral blood DNA of the child and his parents were analyzed by whole exome sequencing. Candidate variants were validated by Sanger sequencing, followed by bioinformatics analysis, American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines for the Interpretation of Sequence Variants pathogenicity classification, and protein structure prediction. A literature search with "Coats Plus syndrome" or "Cerebroretinal microangiopathy with calcifications and cysts" as keywords was conducted at PubMed, China National Knowledge Infrastructure and Wanfang databases to include recently published studies (up to December 2023). This study has been approved by the Ethics Committee of Hunan Children's Hospital (Ethics No. KY2020-07). Informed consent for clinical research was obtained from the guardian of the child. RESULTS: The proband was a 10-year-10-month-old boy. The clinical manifestations were intrauterine and postnatal growth retardation, gastrointestinal hemorrhage, liver fibrosis, panhemopenia, bilateral exudative retinopathy, intracranial lesions and facial pigmentation. WES and Sanger sequencing revealed two novel heterozygous variants in the CTC1 gene: c.787G>A (p.Val263Met) in exon 5 and c.2930C>G (p.Ser977Cys) in exon 17, which were inherited from his mother and father, respectively. According to ACMG pathogenicity classification, both missense variants were classified as variants of uncertain significance (VUS). Protein structure prediction showed the absence of LIG_SH3_3 motif and LIG_SH3_3 motif, and the p.Ser977Cys mutation may affect the binding between CST (CTC1-STN1-TEN) complex and DNA strand. The child had continued to experience recurrent gastrointestinal bleeding episodes despite propranolol treatment, but the condition was controlled after liver transplantation. According to the predefined literature search strategy of this study, a total of 10 relevant articles on pediatric CRMCC patients were retrieved, involving 11 children with gastrointestinal bleeding. Pharmacological and endoscopic therapies play a certain role in the management of CRMCC children complicated with gastrointestinal bleeding. CONCLUSION The CTC1 gene c.787G>A and c.2930C>G variants probably underlay CRMCC in this child. This study has broadened the variation spectrum of CTC1-related diseases and provided a basis for genetic counseling. Liver transplantation may be an important treatment for gastrointestinal hemorrhage in children who do not respond well to medication and endoscopic therapy.
Humans ; Male ; Gastrointestinal Hemorrhage/genetics* ; Child ; Calcinosis/genetics* ; Cysts/genetics* ; Central Nervous System Cysts/genetics* ; Mutation ; Exome Sequencing ; Leukoencephalopathies ; Retinal Diseases ; Seizures ; Muscle Spasticity ; Brain Neoplasms ; Ataxia

Objective: To explore the effect of lncRNA ADPGK-AS1 on the proliferation and apoptosis of retinoblastoma cells and its possible mechanism. Methods: The tumor tissues of 31 patients with retinoblastoma admitted to Henan Provincial Eye Hospital from February to June 2020 and their corresponding normal tissues adjacent to the cancer were collected. The expression levels of lncRNA ADPGK-AS1 and miR-200b-5p in retinoblastoma tissues and normal adjacent tissues were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Human retinal epithelial cell ARPE-19, human retinoblastoma cell Y-79 and WERI-Rb-1 were cultured in vitro. The expression levels of lncRNA ADPGK-AS1 and miR-200b-5p were detected by qRT-PCR. Y-79 cells were randomly divided into si-con group, si-lncRNA ADPGK-AS1 group, miR con group, miR-200b-5p group, si-lncRNA ADPGK-AS1+ anti-miR con group, and si-lncRNA ADPGK-AS1+ anti-miR-200b-5p group. The proliferation, cloning and apoptosis of cells in each group were detected by tetramethylazol blue method, plate cloning test and flow cytometry, respectively. The targeting relationship between lncRNA ADPGK-AS1 and miR-200b-5p was detected by double luciferase report test, and the expression level of cleaved-caspase-3 protein was detected by western blot. Results: Compared with the adjacent tissues, the expression of lncRNA ADPGK-AS1 in retinoblastoma tissues was increased (P<0.05), while the expression of miR-200b-5p was decreased (P<0.05). Compared with ARPE-19 cells, the expression of lncRNA ADPGK-AS1 in Y-79 and WERI-Rb-1 cells was increased (P<0.05), while the expression of miR-200b-5p was decreased (P<0.05). Compared with the si-con group, the cell viability of the si-lncRNA ADPGK-AS1 group was reduced (1.06±0.09 vs 0.53±0.05, P<0.05), the number of cell clone formation was reduced (114.00±8.03 vs 57.00±4.13, P<0.05), while the apoptosis rate [(7.93±0.68)% vs (25.43±1.94)%] and the protein level of cleaved-caspase-3 were increased (P<0.05). Compared with the miR-con group, the cell viability of the miR-200b-5p group was decreased (1.05±0.08 vs 0.57±0.05, P<0.05), the number of cell clone formation was decreased (118.00±10.02 vs 64.00±5.13, P<0.05), while the apoptosis rate [(7.89±0.71)% vs (23.15±1.62)%] and the protein level of cleaved-caspase-3 were increased (P<0.05). lncRNA ADPGK-AS1 could target the expression of miR-200b-5p. Compared with the si-lncRNA ADPGK-AS1+ anti-miR-con group, cell viability of the si-lncRNA ADPGK-AS1+ anti-miR-200b-5p group was increased (0.53±0.04 vs 1.25±0.10, P<0.05), and the number of cell clones was increased (54.00±4.39 vs 125.00±10.03, P<0.05), while the rate of apoptosis [(25.38±1.53)% vs (9.76±0.71)%] and the protein level of cleaved-caspase-3 were decreased (P<0.05). Conclusion: Interfering with the expression of lncRNA ADPGK-AS1 could inhibit the proliferation and clone formation and induce apoptosis of retinoblastoma cells by targeting the expression of miR-200b-5p.
Humans ; MicroRNAs/metabolism* ; Retinoblastoma/pathology* ; Caspase 3/metabolism* ; RNA, Long Noncoding/metabolism* ; Antagomirs/pharmacology* ; Cell Proliferation ; Cell Line, Tumor ; Apoptosis/genetics* ; Retinal Neoplasms/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Movement/genetics*

OBJECTIVETo investigate the expression of ALDH1, CXCR4 and E-cadherin in gastric carcinoma and their roles in lymphatic metastasis.

METHODSSurgical specimens from 127 cases of gastric carcinoma were examined for expressions of ALDH1, CXCR4 and E-cadherin immuohistochemistry with 60 adjacent tissues as control. The associations of ALDH1, CXCR4 and E-cadherin with the clinicopathological pfeatures, 5-year survival rate and lymph node metastasis of the patients were analyzed.

RESULTSALDH1, CXCR4 and E-cadherin were positive in 57.5% (73/127), 63.8% (81/127), and 36.2% (46/127) of the gastric carcinoma tissues, respectively, showing significant differences from the rates in the adjacent tissues (P<0.05). The expression of ALDH1 was significantly correlated with TNM stage and lymph node metastasis (P<0.05), CXCR4 was significantly correlated with the invasion depth, differentiation, TNM stage and lymph node metastasis of the tumor (P<0.05), and E-cadherin was significantly correlated with the invasion depth, differentiation and lymph node metastasis (P<0.05). The positivity rates of ALDH1, CXCR4 and E-cadherin were higher in cases with lymph node metastasis than in those without metastasis. E-cadherin expression was inversely correlated with ALDH1 and CXCR4 expression, and the latter two were positively correlated (P<0.001). Overexpressions of ALDH1 and CXCR4 and a decreased expression of E-cadherin were all related to a poor prognosis of the patients (P<0.05). The expressions ofALDH1, CXCR4 and E-cadherin were all independent prognostic factors of gastric carcinoma.

CONCLUSIONThe expressions of ALDH1, CXCR4 and E-cadherin are associated with the invasion, metastasis and prognosis of gastric carcinoma, and their combined detection provides important evidence for predicting the progression and prognosis of gastric carcinoma.

Cadherins ; genetics ; metabolism ; Carcinoma ; genetics ; metabolism ; Disease Progression ; Humans ; Isoenzymes ; genetics ; metabolism ; Lymphatic Metastasis ; Prognosis ; Receptors, CXCR4 ; genetics ; metabolism ; Retinal Dehydrogenase ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Survival Rate

The risk of radiotherapy-related secondary cancers in children with constitutional retinoblastoma 1 (RB1) mutations has led to reduced use of external beam radiotherapy (EBRT) for RB. Presently, tumor reduction with chemotherapy with or without focal surgery (chemosurgery) is most commonly undertaken; EBRT is avoided as much as possible and is considered only as the last treatment option prior to enucleation. Nevertheless, approximately 80% of patients are diagnosed at a locally advanced stage, and only 20-25% of early stage RB patients can be cured with a chemosurgery strategy. As a whole, chemotherapy fails in more than two-thirds of eyes with advanced stage disease, requiring EBRT or enucleation. Radiotherapy is still considered necessary for patients with large tumor(s) who are not candidates for chemosurgery but who have visual potential. When radiation therapy is indicated, the lowest possible radiation dose combined with systemic or local chemotherapy and focal surgery may yield the best clinical outcomes in terms of local control and treatment-related toxicity. Proton beam therapy is one EBRT method that can be used for treatment of RB and reduces the radiation dose delivered to the adjacent orbital bone while maintaining an adequate dose to the tumor. To maximize the therapeutic success of treatment of advanced RB, the possibility of integrating radiotherapy at early stages of treatment may need to be discussed by a multidisciplinary team, rather than considering EBRT as only a last treatment option.
Child ; Child, Preschool ; Eye Neoplasms/genetics ; Genes, Retinoblastoma/genetics ; Humans ; Radiotherapy Dosage ; Retinal Neoplasms/*radiotherapy ; Retinoblastoma/genetics/*radiotherapy

PURPOSE: Recent reports suggest the association of human papilloma virus (HPV) with retinoblastoma. This study was performed to elucidate whether HPV infection is related to retinoblastoma among Koreans. METHODS: A total of 54 cases diagnosed with retinoblastoma were enrolled from Seoul National University Children's Hospital and Seoul Metropolitan Government-Seoul National University Boramae Medical Center. Presence of human papilloma viral DNA was detected by in situ hybridization in formalin-fixed paraffin-embedded retinoblastoma tissues using both probes against high- and low risk HPV types. RESULTS: The mean age at diagnosis was 22.0 months (range, 1.1 to 98.0 months), and the mean age at enucleation was 27.8 months (range, 1.5 to 112.7 months) among the 54 patients with retinoblastoma. HPV was not detected in any of the retinoblastoma samples using either high risk or low risk HPV probes. CONCLUSIONS: Our study, being the first study in the Korean population, proposes that HPV infection may have no causal relationship with retinoblastoma in Koreans.
Child, Preschool ; DNA, Viral/*analysis ; Eye Infections, Viral/complications/diagnosis/*epidemiology ; Female ; Humans ; In Situ Hybridization ; Incidence ; Infant ; Male ; Papillomaviridae/*genetics ; Papillomavirus Infections/complications/diagnosis/*epidemiology ; Prevalence ; Prognosis ; Republic of Korea/epidemiology ; Retinal Neoplasms/complications/pathology/*virology ; Retinoblastoma/pathology/*virology

OBJECTIVEIdentification of Rb1 mutations permits accurate genetic counseling. Characterization of the causative mutation in a large low penetrance family is likely to provide important information for tumorigenesis of retinoblastoma(RB).

METHODSQuantitative fluorescent multiplex PCR QFM-PCR technique was used for mutation detection. Long fragment PCR, reverse transcriptase-PCR, subcloning, direct sequencing and Western blotting techniques were used for characterizing the mutation.

RESULTSA deletion covering exons 24 and 25 of Rb gene was found in a large family with 122 members in four generations. Of the 18 carries in the family, only 11 were delivered to either unilateral or bilateral RB. The family has much low-penetrance retinoblastoma, compared with the usual, high-penetrance RB (95%). An extent of 4 kb fragment deletion was detected in genomic deletion of the mutation. cDNA and sequence data showed a 174 bp shorter than the wild type message RNA resulting in an in-frame loss of 58 residues. Further analysis demonstrated the truncated protein expression of 6000 Da shorter than wild type RB1 protein.

CONCLUSIONQFM-PCR technique has enabled the investigators to identify a large deletion covering entire exons 24 and 25 of the Rb1 gene. It is the largest deletion ever found in low penetrance RB families. The characterizations of the mutation in genomic DNA, RNA and protein have provided new evidences which enhance credence to the idea that low penetrance retinoblastoma is caused by only partially functional disable of Rb1. The data will be useful in genetic counseling, particularly significant for the unaffected carriers in RB low penetrance families.

Exons ; Female ; Gene Deletion ; Gene Expression ; Humans ; Male ; Pedigree ; Penetrance ; Retinal Neoplasms ; genetics ; Retinoblastoma ; genetics ; Retinoblastoma Protein ; genetics