[Advances on gene therapy for USH2A exon 13 related inherited retinal dystrophy].

Biallelic pathogenic variants in the USH2A gene result in Usher syndrome type Ⅱ and non-syndromic retinitis pigmentosa, both of which entail the progressive loss of photoreceptors leading to blindness. The cDNA of the USH2A gene is extensive, consisting of 15 606 base pairs, rendering it impractical for delivery via adeno-associated virus vectors for gene replacement therapy. Notably, exon 13 has emerged as a focal point for therapeutic intervention, given its predilection for harboring the most pathogenic variants within USH2A. Recent intervention studies targeting USH2A exon 13 through the utilization of antisense oligonucleotides, genome editing, and RNA editing approaches have exhibited promising therapeutic potential. This paper provides a comprehensive overview of the molecular mechanisms, outcome data, and the challenges associated with the application of these interventions in this domain.

[Full-field stimulus threshold testing for evaluation of the retinal function in patients with inherited retinal dystrophy].

Objective: To analyze the characteristics of full-field stimulus thresholds (FST) in patients with middle-and late-stage inherited retinal dystrophy (IRD), and to explore the feasibility of the FST test in evaluating the retinal function for IRD patients. Methods: It was a retrospective case series study. Twenty-eight patients with IRD whose full-field electroretinogram (ERG) results showed no electrical activity (20 patients) or a serious decrease in electrical activity (8 patients) were enrolled, including 17 males and 11 females, with a median age of 32 years. Ten normal controls were also enrolled. All eyes had a measurement of best corrected visual acuity (BCVA) and an ERG test. FST stimulated by red, blue and white lights were examined with an electrophysiological instrument. The decimal value of BCVA was converted to logarithm of the minimum angle of resolution. Data of the better eye of each participant were used for statistical analysis. Independent sample t-test was used to analyze the difference in FST among patients. Linear regression was used to analyze the correlation between FST, visual acuity and ERG amplitudes. Results: The BCVA of all eyes was 0.7±0.6. Under the stimulation of red, blue and white lights, the FST were(-27.0±7.1), (-47.4±12.2) and(-41.7±11.5) dB in the 8 eyes with decreased electrical activity, (-16.3±7.0), (-27.2±13.7) and(-23.5±12.5) dB in the 20 eyes with no electrical activity, and(-39.9±4.0), (-65.8±4.0) and(-58.5±3.4) dB in the 10 control eyes, respectively. The FST of IRD patients were higher than the normal controls. The FST with red, blue and white lasers in eyes with no electrical activity were higher than those with severely reduced electrical activity (t=-3.472, -3.506, -3.433; all P=0.002). The BCVA was not correlated with the FST under the red, blue and white light stimulation in eyes with a serious reduction in electrical activity (r=0.134, 0.011, 0.010; P=0.055, 0.601, 0.611). There was a linear correlation between the red light stimulus FST and the b-wave amplitude of flicker ERG response (r=-0.591, P=0.026), but there was no significant correlation between other amplitudes and FST. Conclusions: The FST test is a supplementary method to evaluate the retinal function of patients with middle-and late-stage IRD. It can quantitatively reflect the function of rod and cone cell system. As the retinal function of IRD patients decreases, the FST gradually increase.

[Correctly interpret the genetic testing results of inherited optic neuropathy].

Inherited optic neuropathy (ION), which can be characterized by isolated optic atrophy or together with systemic manifestations, is a rare optic nerve dysfunction with multiple genetic patterns. Genetic testing plays the key roles in the diagnosis, genetic counseling and treatment of ION. Currently, most ophthalmologists are unfamiliar with how to interpret the genetic testing results correctly. This article describes the genetic characteristics of ION and explains the main points on genetic reports interpretation to help ophthalmologists getting familiar with this relatively rare field and to promote the clinical application of gene testing. (Chin J Ophthalmol, 2021, 57: 326-330).

UCHL1 enhances the malignant development of glioma via targeting GAS2.

OBJECTIVE: To detect the expression pattern of UCHL1 in glioma samples and its influence on the metastasis of glioma, as well as the underlying mechanism. PATIENTS AND METHODS: UCHL1 levels in 42 paired glioma tissues and paracancerous ones were detected. The relationship between UCHL1 level and pathological indexes in glioma patients was analyzed. After establishing UCHL1 knockdown model in U251 and T98-G cells, their migratory ability was assessed by transwell and wound healing assay. At last, Luciferase assay, Western blot and rescue experiments were conducted to explore the role of UCHL1 in aggravating the development of glioma through targeting GAS2. RESULTS: UCHL1 was upregulated in glioma samples than paracancerous ones. High level of UCHL1 indicated high rates of lymphatic metastasis and distant metastasis, as well as low rates of overall survival and progression-free survival in glioma. Knockdown of UCHL1 markedly inhibited migratory ability in glioma cells. GAS2 was the downstream gene of UCHL1. A positive correlation was found between expression levels of UCHL1 and GAS2 in glioma tissues. Overexpression of GAS2 could reverse the inhibitory effects of silenced UCHL1 on migratory ability in glioma cells. CONCLUSIONS: UCHL1 level is linked to lymphatic metastasis, distant metastasis and prognosis in glioma patients. It stimulates migratory ability in glioma by positively regulating GAS2 level.

Up-regulation of long non-coding RNA MFI2 functions as an oncogenic role in cervical cancer progression.

OBJECTIVE: Cervical cancer is one of the gynecologic tumors in the world. The main aim of this study was to elucidate the functional role of MFI2 in cervical cancer and provide novel insight into biomarkers and therapeutic strategies for cervical cancer. PATIENTS AND METHODS: The relative expression level of MFI2 was examined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Cell counting kit-8 (CCK-8) assay was involved to determine the ability of cell proliferation. Flow cytometric analysis was performed to detect cell apoptosis. Transwell assay and Matrigel assay were involved to determine cell migration and invasion. Expressions of protein kinase B (AKT), phosphorylated-AKT (p-AKT), B-cell lymphoma-2 (BCL2), and BCL2-Associated X (Bax) protein levels were detected in Western blotting. Transfected cells were used to perform tumor xenograft formation assay. RESULTS: Our research validated that MFI2 was up-regulated in cervical cancer by qRT-PCR. Through CCK-8 assay, flow cytometric analysis, transwell assay, and Matrigel assay, we verified that MFI2 can promote cell proliferation, cell metastasis and inhibit cell apoptosis in cervical cancer. Subsequently, we used Western blotting assay to determine the alteration of protein expression of p-AKT, BCL2, and Bax. The results indicated that MFI2 may exert its function by regulating phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway. In tumor xenograft formation assay, up-regulated MFI2 accelerated tumor formation. CONCLUSIONS: Current research elucidated that MFI2 promoted cell proliferation, cell metastasis and inhibited cell apoptosis in cervical cancer by regulating the PI3K/AKT signaling pathway. Our results may provide a novel insight into finding new therapeutic targets and biomarkers for cervical cancer.

Aortic dissection presenting primarily as acute spinal cord damage: a case report and literature review.

Acute aortic dissection is a rare, life-threatening condition. Clinical manifestations generally include the acute onset of severe chest or back pain. Aortic dissection presenting with signs and symptoms of acute spinal cord damage is the most severe complication and is particularly rare. This paper reports a case of aortic dissection in a 50-year-old man with a 10-year history of hypertension manifesting as acute spinal cord damage (bilateral lower extremity weakness and loss of all types of sensation), acute skeletal muscle ischaemic necrosis with increased levels of creatine kinase, and acute kidney failure with increased levels of serum creatinine and decreased glomerular filtration rate. The patient refused surgical treatment. His clinical condition progressively worsened and he died 3 days later. This case indicates the importance of considering aortic dissection in patients presenting with acute spinal cord damage, acute skeletal muscle necrosis or acute kidney failure, which may allow early diagnosis and reduce the mortality rate.

Unique phenotype in a Chinese family pedigree: ectopia lentis with varicose great saphenous vein.

OBJECTIVE: To report a Chinese family affected with both ectopia lentis and varicose great saphenous vein. DESIGN: Observational pedigree report. PARTICIPANTS: The family with a total of 53 members in five generations. In the kindred there were 16 affected adults (including 6 deceased), of which 7 were male and 9 were female. MAIN OUTCOME MEASURES: Patients in this family showed an autosomal dominant trait of ectopia lentis and varicose great saphenous vein, occurring in four successive generations. The onset ages for lens dislocation were between 38 and 52 years. No cardiovascular abnormality was observed. Four patients underwent intracapsular lens extraction surgery. CONCLUSIONS: The phenotype of this family showed similarities with Marfan-related disorders. This is a unique phenotype of ectopia lentis with varicose great saphenous vein.

The functional effect of pathogenic mutations in Rab escort protein 1.

Choroideremia (CHM) is a chorioretinal degeneration with an X-linked pattern of inheritance. Affected males experience progressive atrophy of the choroid, retinal pigment epithelium and retina leading to eventual blindness. The CHM gene encodes Rab escort protein 1 (REP-1). REP-1 is involved in trafficking of Rab proteins in the cell. To date, the majority of reported mutations in the CHM gene cause a complete loss of REP-1 function. Here we report pathogenic mutations: a novel missense mutation, L550P; a truncation c.1542T>A, STOP; and two deletions (c.525_526delAG and c.1646delC) in the CHM gene and their phenotypic effect. To analyze the effect of mutations, the 3D structure of human REP-1 and the proteins associated with REP-1 function were modeled using sequence homology with rat proteins. In silico analysis of the missense mutation L550P suggests that the proline residue at position 550 destabilizes the beta-structural elements, and the REP-1 tertiary structure. Truncation and deletion mutants are associated with a partial or total loss of the REP-1 essential activity and protein-protein interactions as predicted by the analysis of the structure and stability of these protein products. The presumptive loss of protein was confirmed by Western Blot analysis of protein from mononuclear cells and fibroblasts (FB) from CHM patients.

Molecular characterization and ophthalmic investigation of a large family with type 2A Von Hippel-Lindau Disease.

BACKGROUND: Von Hippel-Lindau (VHL) disease is a dominantly inherited cancer syndrome. Since the identification of the VHL gene, at least 3 clinical-genetic subtypes of the disease have been recognized. OBJECTIVES: To identify the specific abnormality in the VHL gene and to correlate it with the prevalence and severity of ocular involvement in a large family with VHL disease. METHODS: A longitudinal clinical study and DNA analysis of 24 family members. RESULTS: All 14 affected family members exhibited a thymine-to-cysteine change at nucleotide 505 (T505C) in exon 1 of the VHL gene, consistent with the clinical diagnosis of VHL disease subtype 2A. Two asymptomatic gene carriers were also identified. Seventy-five percent (12/16) of the gene carriers had 1 or more ocular angiomas. The mean number of ocular angiomas per gene carrier was 3.3. Six eyes had optic disc angioma. Five gene carriers (31%) had lost vision because of angiomatosis. Cerebellar hemangioblastomas were present in 4 patients (25%) and pheochromocytomas in 11 (69%). No patient was found to have a renal cell carcinoma. CONCLUSIONS: The family shows a low susceptibility to renal carcinoma consistent with the clinical diagnosis of VHL disease type 2A. The prevalence and severity of ocular angiomatosis in this subtype do not significantly differ from those of the other more common subtypes of VHL. Recognition of the VHL disease 2A phenotype suggests the presence of a specific mutation (T505C) in the VHL gene. Confirmation of this genotype increases the clinician's ability to provide favorable prognostic information to affected family members.

[Prevalence of cataract and surgical coverage among adults aged 50 or above in Shunyi District of Beijing, China].

OBJECTIVE: To estimate the prevalence of cataract, surgical coverage rate and the bilateral cataract-blindness burden among adults aged 50 or above in Shunyi District of Beijing, China, for evaluating the cataract status and the efficacy of the prevention blindness program in more than 10 years. METHODS: Cluster sampling was used in randomly selected individuals aged 50 years or older in 28 villages in Shunyi District in fall, 1996. 5 084 individuals received visual acuity test and eye examination, including lens examination by slit lamp. The cataract operation status was also evaluated. RESULTS: The prevalence of cataract was 23.31% among adults aged 50 or above. The prevalence of cataract was increasing by aging. The prevalence of cataract in women and illiterates was significantly higher statistically. The cataract surgical rate had been increasing since 1984. When the bilateral pinhole vision < 3/60 was defined as the blindness criterion, the cataract blindness surgical coverage rate was 56.36%, and the bilateral cataract-blindness burden was 1.63%. When the bilateral presenting vision < 6/60 was defined as the blindness criterion, the cataract blindness surgical coverage rate was 47.79%, and the bilateral cataract-blindness burden was 2.22%. The cataract surgical coverage rates were much lower and the bilateral cataract-blindness burden much higher in women, illiterates and persons aged 70 or above. CONCLUSIONS: The prevalence of cataract is higher and bilateral cataract-blindness burden is heavier in elder individuals. Cataract blindness is still a serious public health problem in elder, women and illiterates after the prevention blindness program having been actively conducted for more than ten years.

Refractive Error Study in Children: results from Shunyi District, China.

PURPOSE: To assess the prevalence of refractive errors and vision impairment in school-age children in Shunyi District, northeast of Beijing, the Peoples Republic of China. METHODS: Random selection of village-based clusters was used to identify a sample of children 5 to 15 years of age. Resident registration books were used to enumerate eligible children in the selected villages and identify their current school. Ophthalmic examinations were conducted in 132 schools on children from 29 clusters during May 1988 to July 1998, including visual acuity measurements, cycloplegic retinoscopy, cycloplegic autorefraction, ocular motility evaluation, and examination of the external eye, anterior segment, media, and fundus. Independent replicate measurements of all children with reduced vision and a sample of those with normal vision were done for quality assurance monitoring in three schools. RESULTS: A total of 6,134 children from 4,338 households were enumerated, and 5,884 children (95.9%) were examined. The prevalence of uncorrected, presenting, and best visual acuity 0.5 (20/40) or worse in at least one eye was 12.8%, 10.9%, and 1.8%, respectively; 0.4% had best visual acuity 0.5 or worse in both eyes. Refractive error was the cause in 89.5% of the 1,236 eyes with reduced vision, amblyopia in 5%, other causes in 1.5%, with unexplained causes in the remaining 4%. Myopia -0.5 diopter or less in either eye was essentially absent in 5-year-old children, but increased to 36.7% in males and 55.0% in females by age 15. Over this same age range, hyperopia 2 diopters or greater decreased from 8.8% in males and 19.6% in females to less than 2% in both. Females had a significantly higher risk of both myopia and hyperopia. CONCLUSIONS: Reduced vision because of myopia is an important public health problem in school-age children in Shunyi District. More than 9% of children could benefit from prescription glasses. Further studies are needed to determine whether the upward trend in the prevalence of myopia continues far beyond age 15 and whether the development of myopia is changing for more recent birth cohorts.

[The effect of three kinds of anti-glaucoma eyedrops on ocular blood flow].

OBJECTIVE: Surveying the changes of ocular blood flow by continuous anti-glaucoma eyedrops for one week. METHODS: Thirty-two patients in glaucoma clinic were divided into three groups, one treated by 0.5% timolol, one by 0.5% levobunolol and the third by 0.1% dipivefrine, twice a day. Each patient got the retinal and choroidal angiography before and after the medication. We also made the intra- and inter-observer reproducibility test for verifying the credibility of the method used to determine the ocular blood flow. All the data were analyzed by Students't test. RESULTS: The reproducibility test revealed this method creditable. Timolol and levobunolol could decrease the choroidal blood flow significantly, there was a trend to decrease the optic nerve blood flow after continues application of timolol eyedrops. Dipivefrine did not induce any significant change on ocular blood flow except the mild artery constriction. CONCLUSION: The results suggest that it is necessary to be careful in choosing the anti-glaucoma eyedrops when ischemia plays an important role in glaucomatous damage.

[Evaluating retinal nerve fiber layer by scanning laser ophthalmoscopy].

OBJECTIVE: To evaluate the efficiency of scanning laser ophthalmoscopy (SLO) on detecting retinal nerve fiber layer defects (RNFLD). METHODS: 95 eyes with primary open angle glaucoma, 37 ocular hypertension, 83 glaucomatous suspect and 34 normal eyes were investigated by SLO to evaluate the retinal nerve fiber layer (RNFL). Recording tapes of 68 eyes were re-investigated by two investigators for estimating the intra- and inter-observer agreement. RESULTS: The intra- and inter-observer agreement of the presence and types of RNFLD was fairly good (Kappa values were 0.66 - 0.76 and 0.59 - 0.65 respectively). The sensitivity and specificity in detecting RNFLD by SLO were 80.0% and 94.1% respectively. CONCLUSION: The evaluation of RNFL by SLO is a quick, accurate and safe method. It may be clinically useful in the diagnosis of glaucoma.

[Prevalence of blindness among adults aged 50 years or above in Shunyi county of Beijing].

OBJECTIVE: To estimate the prevalence of blindness among older adults aged >or= 50 years in Shunyi county of Beijing, China. METHODS: Cluster sampling was used in randomly selecting individuals aged >or= 50 years in 28 villages in Shunyi county in fall, 1996. The survey was preceded by a pilot study where operational methods were refined and quality assurance evaluation was carried out. RESULTS: 5,555 individuals were enumerated. The response rate was 91.5%. In this population, the prevalence of blindness was 1.7% defined as pinhole visual acuity < 0.05 in both eyes; or 2.8% were blind, defined as presenting visual acuity < 6/60 in both eyes. Blindness was associated with older age and female gender. The principal cause of blindness in eyes was cataract. CONCLUSIONS: The prevalence of blindness was decreased 35.8% after ten year active eye-care program in the survey area. However, cataract blindness continues as a significant problem among elderly, especially females, in this population sample of rural area.

Prevalence of blindness and cataract surgery in Shunyi County, China.

PURPOSE: To estimate the prevalence of blindness and cataract surgery among older adults in rural China. METHODS: Cluster sampling was used in randomly selecting men and women aged 50 years or older for visual acuity testing and an eye examination in 28 villages in Shunyi County. The survey, which was carried out in the fall of 1996, was preceded by a pilot study in which operational methods were refined and quality assurance evaluations carried out. RESULTS: Of 5,555 enumerated subjects > or =50 years of age, 91.5% (5,084/5,555) were examined and 90.9% (5,052/5,555) were tested for visual acuity. In this population, 2.8% (139/5,052) were blind, defined as presenting visual acuity less than 6/60 in both eyes. Blindness was associated with older age and female sex. Cataract was the principal cause of blindness in at least one eye in 48.2% (67/139) of blind people. The ratio of those blind from cataract who were operated on to the those who could have been operated on, cataract surgical coverage, was estimated to be 47.8% (54/113). Cataract surgery was associated with younger age but not sex or education. CONCLUSIONS: Blindness, particularly blindness related to cataract, continues to be a significant problem among the elderly, especially women, in this population-based sample of rural Chinese. Despite an active eye-care program in Shunyi County, only half of those who might benefit from cataract surgery are receiving it.

Visual acuity and quality of life outcomes in patients with cataract in Shunyi County, China.

PURPOSE: To measure visual acuity and vision-related quality of life in individuals in rural China operated on for cataract. METHODS: Five thousand fifty-two persons age 50 years and older, 90.9% (5,052/5,555) of a randomly selected population in Shunyi County, were examined in the fall of 1996. Visual functioning and quality of life questionnaires were administered to those with presenting visual acuity less than 6/60 in either eye and to those who were aphakic or pseudophakic. RESULTS: Of the 87 individuals operated on for cataract, 12% (10/87) had presenting visual acuity of 6/18 or more in both eyes, and 24.1% (21/87) had less than 6/60. Twenty-five percent (29/116) of the 116 eyes operated on for cataract had presenting visual acuity of 6/18 or more, and 44.8% (52/116) had less than 6/60. Aphakic cases without glasses and uncorrectable aphakia attributable to surgical complications were common. In a multivariate regression model, including time period of surgery, hospital type, and surgical procedure, only pseudophakia was associated with better outcomes (P = .05). On a scale from 0 (maximum problems) to 100 (no problems), the mean visual functioning score (+/-SD) for the operated-on population was 61.9 +/- 30.0, and 71.0 +/- 31.8 for the quality of life questionnaire. These scores were comparable to those of the unoperated-on population with moderate bilateral blindness (<6/60 to > or =3/60 in the better eye). Visual functioning and quality of life scores were closely correlated with visual acuity in operated-on (r = 0.64 and r = 0.61, respectively) and unoperated-on populations (r = 0.68 and r = 0.59, respectively). CONCLUSIONS: Both clinical and patient-reported cataract surgery outcomes are below what should be achievable. Improvement in outcomes must be given greater emphasis if the potential of cataract surgery in restoring sight is to be realized.

[Clinical application and observation of silicone oil in complicated retinal detachment].

Nine eyes with complicated retinal detachment underwent vitrectomy and silicone oil tamponate among whom there were seven eyes with PVRC3-PVRD2, one eye with giant retina tear and PVRD3 and one eye with giant retina tear. All the eyes were followed-up for an average of 12.6 months with a range from 2 to 23 months. Retina were completely reattached in there eyes. Vision were improved in six eyes. Retina was redetached in one eye after removal of silicone oil. Silicone oil tamponate can be used in complicated retinal detachment that fails to respond to conventional method. However, silicone oil tamponate may cause a number of complications, so its indications should be properly taken care of.

Surgical management of macular epiretinal membranes.

PURPOSE: To evaluate the effects of surgical treatment on macular epiretinal membranes. METHODS: Vitrectomy and membrane removal were undergone for idopathic secondary macular epiretinal membrane. RESULTS: Fourteen eyes of 15 patients (93%) had vision improvement after operation in which 8 eyes (57%) increased 3 or more Senellen lines. Only one case suffered from paracentral scotoma. No other complications were noted. CONCLUSION: Surgical management of macular epiretinal membrane is safe and effective with good visual outcome and few complications.

[Studies on the anticancer effect of HH07A, a derivative of hainanensine].

The effect of HH07A on the growth of tumor cells in vitro was investigated using the techniques of cell growth curve determination and soft-agar colony-forming assay. The result showed that HH07A inhibited the growth of L1210 cells and HL-60 cells at a concentration of 1.5 micrograms.ml-1 and 4.0 micrograms.ml-1, respectively. Among the cells tested, L1210 cells were shown to be the most sensitive, followed by KB cells and HL-60 cells (with IC50 of 2.29, 4.13 and 4.36 micrograms.ml-1, respectively). Normal mouse granulocyte-macrophage progenitor cells (GM-CFC) were less sensitive to the drug (with IC50 of 11.15 micrograms.ml-1) as compared with the tumor cells. As they were exposed to HH07A 3.5 micrograms.ml-1 for 5 days, HL-60 cells did not show NBT reductive ability. Intraperitoneal injection of HH07A exerted inhibitory effect on the ascitic tumors of L1210 and S180 in mice. Oral or ip administration of HH07A also showed some effect on S180 solid tumors in mice.