Advances in the study of POAG-related genes and central nervous system diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#In recent years, considerable progress has been made in the study of glaucoma, especially primary open angle glaucoma(POAG). A series of POAG genes has been identified through genetic linkage analysis and genome-wide association studies(GWAS), which significantly advanced the study of glaucoma genetics. The latest perspective suggests that glaucoma is a disease of the central nervous system(CNS). A large number of basic clinical studies have demonstrated the close association between CNS disease and glaucoma. Among these studies, discoveries related to genetics are of prominence.

Therapeutic effects of oleanolic acid nanoparticles combined with heparin sodium nanoparticles on liver cancer in vivo and in vitro / 西安交通大学学报(医学版)

Objective To investigate the therapeutic effects of oleanolic acid-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate (PLGA-TPGS ) nanoparticles (OPTN ) combined with heparin sodium-loaded PLGA-TPGS nanoparticles (HPTN)on liver cancer,and explore whether OPTN combined with HPTN has a synergistic effect by comparing the results of single medication.Methods Coumarin 6 and eosin were used as fluorescent probes to examine the cellular uptake by human liver cancer HepG2 cell and murine ascitic hepatocarcinoma cell strain with high metastasis potential in the lymphatic system (HCa-F).The in vitro cytotoxic combination effect and apoptosis of HepG2 cells induced by OPTN combined with HPTN were also determined using WST-1 assay and Annexin V-FITC staining. The antitumor effect in vivo was determined by tumor growth inhibition rate and hematoxylin & eosin staining (H & E)method.Results Both OPTN with green fluorescence and HPTN with red fluorescence were found in HepG2 cells and HCa-F cells,suggesting that they had been internalized.The cell cytotoxicity test and Annexin V-FITC staining results showed that OPTN combined with HPTN had a synergistic effect.The therapeutic effect in vivo showed that OPTN combined with HPTN effectively inhibited tumor growth better than the drug alone.Conclusion OPTN combined with HPTN has a synergistic effect in more effectively inhibiting liver cancer better than single medication.

Therapeutic effects of oleanolic acid nanoparticles combined with heparin sodium nanoparticles on liver cancer in vivo and in vitro / 西安交通大学学报(医学版)

Objective To investigate the therapeutic effects of oleanolic acid-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate (PLGA-TPGS ) nanoparticles (OPTN ) combined with heparin sodium-loaded PLGA-TPGS nanoparticles (HPTN)on liver cancer,and explore whether OPTN combined with HPTN has a synergistic effect by comparing the results of single medication.Methods Coumarin 6 and eosin were used as fluorescent probes to examine the cellular uptake by human liver cancer HepG2 cell and murine ascitic hepatocarcinoma cell strain with high metastasis potential in the lymphatic system (HCa-F).The in vitro cytotoxic combination effect and apoptosis of HepG2 cells induced by OPTN combined with HPTN were also determined using WST-1 assay and Annexin V-FITC staining. The antitumor effect in vivo was determined by tumor growth inhibition rate and hematoxylin & eosin staining (H & E)method.Results Both OPTN with green fluorescence and HPTN with red fluorescence were found in HepG2 cells and HCa-F cells,suggesting that they had been internalized.The cell cytotoxicity test and Annexin V-FITC staining results showed that OPTN combined with HPTN had a synergistic effect.The therapeutic effect in vivo showed that OPTN combined with HPTN effectively inhibited tumor growth better than the drug alone.Conclusion OPTN combined with HPTN has a synergistic effect in more effectively inhibiting liver cancer better than single medication.

Construction of shRNA eukaryotic expression vector targeting OPTN gene and detection of interference effect / 安徽医科大学学报

Objective To construct shRNA against optineurin ( OPTN ) clone and transfect the construction into HEK293FT cells for inhibiting the expression of OPTN, which is laid the foundation for further research of molecu-lar mechanism OPTN protein. Methods The insert of this clone was a double-strained DNA sequence against OPTN which would be transcripted into shRNA and it was synthesized according to the sequence in Origene. Ex-pression vector pRFP-C-RS was employed to fuse the insert to get the construction and finally confirmed by sequen-cing. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN and RFP signals could be detected using fluorescence microscope. Western blot was further employed to check the protein level of OPTN. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN was then obtained to investigate the functional study of OPTN which was measured by Salmonella infection experiment. Results shRNA eukaryotic expression vector targeting OPTN gene was successfully constructed. HEK293FT cells were transfected with pRFP-C-RS-shOPTN and 72 hours later we observed strong RFP signals showing that shOPTN expressed. Western blot was further employed to check the protein level of OPTN which showed that OPTN expression was indeed interfered. Salmonella infection assays was then performed and showed that OPTN could inhibit the proliferation of Salmonella that has invaded HEK293FT cells. Conclusion HEK293FT cell line which expresses shRNA against OPTN show a sharp inhibition of OPTN protein expression by 80%. So this cell line can be further used to investigate how OPTN regulates auto-phagy as an autophagy receptor. Meanwhile we find that OPTN can be as a autophagy regulator and strongly sup-press the proliferation of invasive Salmonella in vivo.

Glaucoma in Costa Rica. Initial approaches: [minireview]

Glaucoma is the second most frequent cause of irreversible blindness worldwide. Genetic factors have been implicated in the development of the disease. So far six loci (GLC1A-GLC1F) and two genes (TIGR/MYOC and OPTN) are involved in the development of juvenile (JOAG) and adult onset or chronic primary open angle glaucoma (COAG), while two loci (GLC3A,GLC3B) and one gene (CYP1B1) are known for primary congenital glaucoma (PCG). Here we summarize the results of the first genetic studies of glaucoma in Costa Rica. Nine families: 1 with JOAG, 1 with PCG and 7 with COAG were screened for mutations at the known genes. A 10 bp duplication, 1546-1555dupTCATGCCACC, at the CYP1B1 gene, causes, in homozygous state, glaucoma in the consanguineous PCG family. This mutation has been found in different countries and generates an early stop codon that termitates protein synthesis 140 amino acids earlier than the normal allele. In exon 1 of the T1GR/MYOC the innocuous Arg76Lys variant was found in two of the COAG families. In the OPTN gene two variants in the coding region (Thr34Thr, Met 98Lys) and 7 intronic changes were found in other Costa Rican glaucoma patients. One of the COAG families was chosen for a genome scan with 379 microsatellite markers and linkage analysis. LOD scores [quot ]suggestive[quot ] of linkage were obtained for several chromosomal regions. Evidence indicates that hereditary glaucoma in Costa Rica is highly heterogeneous and that further studies in the country will probably disclose some up to now unknown genes responsible for the disease.

OPTN gene Mutation in Normal-Tension Glaucoma

PURPOSE: We have evaluated the mutations of the OPTN gene, which has been reported to be associated with the normal tension glaucoma (NTG). METHODS: The OPTN gene was analyzed in 53 patients with NTG and 40 normal subjects. Genomic DNA was extracted from the blood samples of each patients, exon 5 and exon 6 of the OPTN gene were amplified by PCR and DNA sequencing was performed. RESULTS: No mutation was found in normal subjects. But three kinds of point mutation (G412A, C459T in exon 5, G577C in exon 6) were found in 7 patients with NTG. CONCLUSIONS: We report the novel point mutations of OPTN gene in NTG patients. This shows the possibility of diagnosis of NTG by detecting the mutation of OPTN gene.