OBJECTIVE: To explore the clinical phenotype, genotype and genetic characteristics for a patient with unilateral Pigmented paravenous retinochoroidal atrophy (PPRCA) and Retinitis pigmentosa (RP) in the contralateral eye. METHODS: A PPRCA pedigree which had presented at the Department of Medical Genetics of the Eye Hospital of Wenzhou Medical University in August 2021 was selected as the study subject. Clinical data of the family members were collected. The proband underwent wide-field fundus photography, wide-field autofluorescence, full-field electroretinogram (ff-ERG), visual field testing, optical coherence tomography (OCT), and fundus angiography (FFA and ICGA). Blood samples were collected from the proband and family members (parents and two sisters), and buccal mucosal cells were collected from the proband's daughter, and genomic DNA was extracted for each family member. Whole exome sequencing (WES) was performed on the proband. Candidate variants were verified using Sanger sequencing and pathogenicity analysis. This study was approved by the Medical Ethics Committee of the Eye Hospital of Wenzhou Medical University (Ethics No. 2019-134). RESULTS: Wide-angle fundus photography and autofluorescence showed that the right eye was consistent with PPRCA and the left eye with RP. OCT showed that the outer layer of the fovea was intact in the right eye, while disorganized outer segment was found in the fovea of the left eye, and outer segment atrophies outside the fovea were found in both eyes. The amplitudes of ff-ERG decreased significantly in both eyes, and the amplitudes in right eye were slightly higher than those of the left eye. Visual field showed a paracentral arcuate scotoma in the right eye and severe centripetal contraction in the left eye. FFA showed hyperfluorescence in the retinal vein distribution area caused by atrophy of retinal pigment epithelium of the right eye and hypofluorescence related to bone spicule pigmentation, in addition with mottled hypofluorescence of choroid in the left eye. ICGA showed mild paravenous retinochroidal atrophy of the right eye and diffuse choroid capillaries atrophy in the middle and peripheral area of the left eye. WES revealed that the proband had a heterozygous c.2234C>T (p.Thr745Met) variant of the CRB1 gene. Sanger sequencing confirmed that the proband and family members except the father of the proband carried the same CRB1 gene variant. Based on the criteria and guidelines for the classification of genetic variation and related consensus from the American College of Medical Genetics and Genomics (ACMG), this variant was classified as pathogenic (PM3_VeryStrong+PM1+PM2_Supporting +PP3). CONCLUSION The heterozygous c.2234C>T (p.Thr745Met) variant of the CRB1 gene may underlay the unilateral PPRCA with contralateral eye RP in this proband. Above findings have enriched the mutational spectrum of the CRB1 gene.
Humans ; Electroretinography ; Exome Sequencing ; Eye Proteins/genetics* ; Membrane Proteins/genetics* ; Mutation ; Nerve Tissue Proteins/genetics* ; Pedigree ; Phenotype ; Retinitis Pigmentosa/genetics* ; Tomography, Optical Coherence ; Retinal Degeneration ; Eye Diseases, Hereditary

BACKGROUND: Magnetic field can affect the growth and division of cancer cells both in vivo and in vitro, however, the effects on apoptosis of human liver cancer cells induced by magnetic field is still unclear.OBJECTIVE: To explore the inducing effect of extremely low fre quency (ELF) magnetic field on apoptosis of human liver cancer cell SK-HEP- 1.JESIGN: An open experiment with cells as the observational subjects.SETTING: Institute of Biotechnology, Shanghai Jiaotong University.MATERIALS: The experiment was carried out at the Institute of Biotechnology, Shanghai Jiaotong University from September 2004 to January 2005. The subject was human liver cancer cell line SK-HEP-1, purchased from cell bank of Chinese Academy of Science, Shanghai.METHODS: SK-HEP-1 cells were inoculated to T-flasks at the density of 2.0×107 cells L-1, and cultivated in the DMEM containing 0.1 volume fraction of heat-inactivated fetal bovine serum and 2 mmol/L L-glutamine. Exposure groups were exposed to 50 Hz, 20 mT magnetic field and the control groups were run concurrently under the same conditions with the exposed cultures but in a separate incubator which was free of magnetic field during 8-day culture process. The apoptosis of SK-HEP-1 cells were defined by DNA ladder assay, Hoechst 33258 staining and AO/EB staining respectively on day 8.MAIN OUTCOME MEASURES: ① DNA fragmentation pattern formation. ② The abnormal nucleus formation. ③ The percentage of apoptotic cells.RESULTS: ① Detection of internucleosomal DNA fragmentation by DNA ladder assay: After 8-day ELF magnetic field exposure, DNA fragmentation pattern was detected by DNA ladder assay, which was not observed in control groups (free of exposure). ② Fluorescence microscopy analysis of apoptosis by Hoechst 33258 staining: Hoechst 33258 staining was used to investigate the changes in the nucleus of cells, and many apoptotic bodies containing nuclear fragments were found in ELF magnetic field exposed cells, but just about none in untreated cells. At the same time, cytoplasmic shrinkage was observed in cells cultured under the exposure. And in some cells, even the cell membrane was unable to keep intact. ③ Fluorescence microscopy analysis of cell apoptosis by acridine orange/ethidium bromide (AO/EB) double staining: After ELF magnetic field exposure, the percentage of viable cells in exposure groups (9.2%) was rather low compared with the control groups (91.8%), and was accompanied with a high apoptotic rate (72.3%), while only 4.2% in control group. A large number of apoptotic cells were at the early stage of apoptosis. The rate of apoptotic cells (18.5%)after treated by magnetic field was higher than that of control group (4%). With the AO/EB double staining, control cells appeared to be round,intact and bright green while some of the exposed cells exhibited irregular cell morphology and condensed nucleus.CONCLUSION: Apoptosis of human liver cancer cell SK-HEP-1 could be induced by 50 Hz, 20 mT magnetic field in vitro.

Objective: To study the effects of extremely low frequency (ELF) magnetic field with fixed parameters on human liver cancer cells (SK-HEP-1) at different aspects. Methods: SK-HEP-1 cells were exposed to 50Hz, 20mT magnetic field during the whole culture process, and then proliferation activity, growth kinetics, metabolic profile and cell cycle were analyzed. Results: 50Hz, 20mT magnetic field inhibits the growth and metabolism of SK-HEP-1 cells, and hampers their mitotic division. Conclusion: 50Hz, 20mT magnetic field could be a potential therapy in the treatment of human malignant tumors.

OBJECTIVETo determine the in vitro expression of interleukin-12 (IL-12) and its effect on signal transducers and activators of transcription (STAT) signaling molecules in peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE).

METHODSPeripheral blood mononuclear cells in 39 patients with definite systemic lupus erythematosus and 11 healthy volunteers were collected. Expression of IL-12 P40mRNA in PBMCs was determined with reverse transcription-polymerase chain reaction (RT-PCR). Quantity of IL-12 protein supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The levels of phosphorylated STAT3 and STAT4 signaling molecules in PBMCs were detected by immunoblot.

RESULTSLevels of IL-12 protein and mRNA expression in patients with active or inactive SLE were significantly higher than those in controls. Phytohemagglutinin (PHA ) may promote the expression of IL-12. IL-12 alone induced the phosphorylation of STAT3 and STAT4 in PBMCs from patients with SLE, especially in active SLE. However it had no obvious effect on normal PBMCs. Phosphorylated STAT3 and STAT4 might be observed in normal PBMCs treated with IL-12 plus PHA.

CONCLUSIONIL-12 is produced aberrantly in patients with SLE. IL-12 might exert its biological role in SLE via the aberrantly phosphorylated STAT3 and STAT4 signaling molecules.

Adolescent ; Adult ; Cells, Cultured ; DNA-Binding Proteins ; metabolism ; Humans ; Interleukin-12 ; blood ; genetics ; Leukocytes, Mononuclear ; metabolism ; Lupus Erythematosus, Systemic ; metabolism ; Middle Aged ; Phosphorylation ; RNA, Messenger ; analysis ; STAT3 Transcription Factor ; STAT4 Transcription Factor ; Trans-Activators ; metabolism