TonoVet versus Tonopen in a high intraocular pressure monkey model.

Purpose: The purpose of this study is to evaluate the consistency of and deviation in intraocular pressure (IOP) measurements obtained by the TonoVet rebound tonometer and the Tonopen applanation tonometer in a primate model. Methods: Twenty-four-hour IOPs (nine time points) were recorded in ten monkeys with normal IOP and eight monkeys with chronic high IOP (one eye was randomly selected for measurement in each animal) using a Tonopen and TonoVet device. Measurements obtained using both handheld devices were first compared in the healthy control group (90 readings). The monkeys with chronic ocular hypertension (COHT, 72 IOP readings) were divided into three subgroups according to the level of IOP. The consistency of and deviations in the measurements were analyzed using Bland-Altman plots, linear regression, and two-tailed Student t tests. Results: In monkeys with normal IOP, the two devices produced similar IOP readings (mean IOP deviation, 0.06 ± 2.08 mmHg, p = 0.761), with 56.67% of the deviation between -1 mmHg and 1 mmHg and 91.12% between -3 mmHg and 3 mmHg. However, in the animal model group (23-60 mmHg), the readings obtained by the TonoVet tonometer were higher than those obtained by the Tonopen tonometer (mean deviation, 13.76 ± 9.19 mmHg); furthermore, 75.68% of the TonoVet measurements deviated by ± 5 mmHg from the Tonopen measurements. Conclusions: In animals with normal IOP, the TonoVet and Tonopen tonometers produced consistent measurements. However, in a monkey model of chronic high IOP, the measurements obtained by these tonometers were inconsistent, with higher IOPs associated with larger measurement errors. Therefore, it is necessary to be aware that among different tonometers, there may be systemic errors and deviations in IOP measurements. These findings should facilitate efforts to obtain more accurate individualized diagnoses and prevent the utilization of misleading IOP values.

Alpha 1-antitrypsin inhibits microglia activation and facilitates the survival of iPSC grafts in hypertension mouse model.

This study was conducted to investigate the use of Alpha 1-antitrypsin (AAT) to inhibit microglia activation in chronic hypertension model and provide a permissive environment for stem cell transplantation. Chronic ocular hypertension of C57BL/6 mice using magnetic microbead injection was induced 3 weeks prior to iPSCs transplantation. The ocular hypertension model was assessed histologically and intraocular pressure was measured. Survival of grafted cells and microglia activation were examined by flow cytometry and immunofluorescence in AAT and PBS treated hosts. Retinal cytokines expression was also detected by real-time PCR. Chronic ocular hypertension resulted in persistent microglia activation and stem cell grafts loss. AAT treatment significantly inhibited microglia activation and facilitated the survival of transplant iPSCs 4w post transplantation compared to PBS treatment. AAT holds tremendous potential for the clinical application to control neuroinflammation factor in glaucoma and improve the stem cell replacement therapy of retinal neurodegenerative disease.

[Effect of Zishen Huoxue Recipe on Pathomorphology in Coronary Heart Disease Rats with Shen Deficiency Blood Stasis Syndrome].

OBJECTIVE: To observe the effect of Zishen Huoxue Recipe (ZHR) on pathomorphology in coronary heart disease (CHD) rats with Shen deficiency blood stasis syndrome (SDBSS). METHODS: Totally 60 healthy Wistar rats were divided into the blank control group, the model group, high, middle, and low dose ZHR groups according to random digit table, 12 in each group. Myocardial ischemia SDBSS rat model was prepared by ligating the left anterior descending coronary artery and injecting hydrocortisone. ZHR physic liquor was administered to rats in high, middle, and low dose ZHR groups at the daily dose of 21.6, 10.8, 5.4 g/kg by gastrogavage for 7 successive days, equal volume of pure water was administered to rats in the blank control group and the model group by gastrogavage for 7 successive days. Rat heart was collected for pathomorphological observation under light microscope. RESULTS: In the model group the heart muscle fiber was swollen and deformed with widened space, loose and dropsy tissues. Blood vessels in myocardial mesenchymal were dilated, infiltrated with more inflammatory cells. Myocardial cells were markedly swollen, degenerated, or necrotic, with caryolysis or disappearance of partial nuclear. A large amount of collagen fibrous tissue became hyperplasia. Endocardial blood vessels were swollen and degenerated with infiltration of few inflammatory cells. Epicardium tissue and structure were destroyed and got hyperplasia. Swollen, degenerated, or necrotic vessels could be seen, with infiltration of more inflammatory cells and collagen deposition. Pathomorphological injuries were alleviated in each ZHR group. The higher ZHR concentration, the milder the injury degree of myocardial tissue, the more limited range of damage. CONCLUSION: ZHR could attenuate pathomorphological injuries of myocardial ischemia rats with SDBSS and regulate myocardial function, thus improving myocardial ischemia in CHD rats with SDBSS.

Mitomycin C-loaded PTMC15-F127-PTMC15 hydrogel maintained better bleb function after filtering glaucoma surgery in monkeys with intraocular hypertension.

There is an unmet need for a safer and more effective approach for antimetabolite application to prevent bleb fibrosis after glaucoma filtering surgery. Here, we utilized our previously developed thermosensitive sustained-release agent, mitomycin C-loaded poly(trimethylene carbonate)15-F127-poly(trimethylene carbonate)15 (MMC-hydrogel), aiming to further evaluate the efficacy and safety of MMC-hydrogel in high intraocular pressure (IOP) primate eyes. Twelve primate eyes after high IOP induction were randomly divided into three groups, which respectively received phosphate-buffered saline (PBS)-hydrogel, MMC-hydrogel, and MMC treatment during trabeculectomy. IOP and bleb volume were measured using a Tonopen and anterior segment optical coherence tomography over 28 days. At the end of the experiment, all experimental primate eyes were enucleated. Histopathology and immunohistochemistry were performed to reveal myofibroblast cells and collagen deposition of filtering blebs. The MMC-hydrogel group had satisfactory IOP control (9.25 ± 4.80 mmHg) and maintained well-functioning blebs for a longer time. Fibrosis and scarring were significantly alleviated in this MMC-hydrogel group. There was no obvious toxicity to ocular surfaces or intraocular structures. Taken together, these data suggest that PTMC15-F127-PTMC15-loaded MMC-hydrogel plays a role in functional maintenance and scarring inhibition, showing high efficacy in reducing post-filtering surgery bleb fibrosis. This MMC-hydrogel may offer a new solution for filtering bleb management after glaucoma surgery.

Biodegradable scaffolds facilitate epiretinal transplantation of hiPSC-Derived retinal neurons in nonhuman primates.

Transplantation of stem cell-derived retinal neurons is a promising regenerative therapy for optic neuropathy. However, significant anatomic differences compromise its efficacy in large animal models. The present study describes the procedure and outcomes of human-induced pluripotent stem cell (hiPSC)-derived retinal sheet transplantation in primate models using biodegradable materials. Stem cell-derived retinal organoids were seeded on polylactic-coglycolic acid (PLGA) scaffolds and directed toward a retinal ganglion cell (RGC) fate. The seeded tissues showed active proliferation, typical neuronal morphology, and electrical excitability. The cellular scaffolds were then epiretinally transplanted onto the inner surface of rhesus monkey retinas. With sufficient graft-host contact provided by the scaffold, the transplanted tissues survived for up to 1 year without tumorigenesis. Histological examinations indicated survival, further maturation, and migration. Moreover, green fluorescent protein-labeled axonal projections toward the host optic nerve were observed. Cryopreserved organoids were also able to survive and migrate after transplantation. Our results suggest the potential efficacy of RGC replacement therapy in the repair of optic neuropathy for the restoration of visual function. STATEMENT OF SIGNIFICANCE: In the present study, we generated a human retinal sheet by seeding hiPSC-retinal organoid-derived RGCs on a biodegradable PLGA scaffold. We transplanted this retinal sheet onto the inner surface of the rhesus monkey retina. With scaffold support, donor cells survive, migrate and project their axons into the host optic nerve. Furthermore, an effective cryopreservation strategy for retinal organoids was developed, and the thawed organoids were also observed to survive and show cell migration after transplantation.

Semi-quantitative analysis for the dynamic chest CT imaging features from onset to recovery in severe and critical COVID-19.

OBJECTIVE: To investigate in the CT manifestations of severe and critical Coronavirus Disease 2019 (COVID-19) patients. METHODS: Medical data was collected for 2 severe patients and 4 critical COVID-19 patients from onset to their recovery. Three or four CT scans for each patient were taken. The semi-quantitative analysis method was introduced for lesion and its distribution area. RESULTS: The ground-glass opacities (GGO) and mixed GGO with consolidation were found as the most frequent features. Consolidation followed, and the appearance of stripes which showed an increasing trend before the patient was discharged. Consolidation was associated with clinical severity and disease progression, and the rapid change of the lesion in a short period of time was also a notable feature within 2-3 weeks. After being discharged, the efficacy of treatment could be demonstrated by a follow up CT scan. The distribution of lesion also showed dynamic progress in the follow up CT scan. CONCLUSION: CT scans in the whole course provided the entire inflammation information to assess clinical severity, disease progression and the treatment efficacy for COVID-19.

Posture-Dependent 24-Hour Intraocular Pressure Fluctuation Patterns in an Intraocular Hypertension Monkey Model.

PURPOSE: We investigate 24-hour intraocular pressure (IOP) fluctuation patterns and the influence of body position on IOP in a chronic ocular hypertension (COHT) monkey model. METHODS: We recorded 24-hour IOPs (nine time points) in the different body positions in 10 eyes with normal and eight with high IOP (with random selection of one eye of each monkey) using a Tonopen. The IOPs at various time points and in different body positions were compared. RESULTS: The average 24-hour IOPs in the immediate-supine, 10-minute supine, 10-minute seated, and immediate-seated positions in the COHT models were 28.64 ± 9.82, 25.42 ± 7.62, 23.49 ± 7.67, and 20.53 ± 7.80 mmHg, respectively. The diurnal-to-nocturnal IOP changes were 8.51 ± 2.93, 5.81 ± 3.67, 5.48 ± 2.97, and 3.59 ± 2.74 mmHg, respectively. The sudden shift between the supine and seated positions bring greater IOP variations (8.11 ± 2.85 mmHg) in the COHT monkeys, and the IOP fluctuations reached 14 to 38 mmHg when considering body position and the measurement time points. CONCLUSIONS: The measurement time and body position influenced IOP. More elevated IOP occurred in the immediate-supine position and during the transient shift between the seated and supine positions. Maintaining a fixed position for sufficient time before measurement is important. TRANSLATIONAL RELEVANCE: Glaucoma patients should focus on the importance of IOP measurements in the clinic occurring after an adequate amount of time in a fixed body position.

Relationship between intraocular pressure and retinal nerve fibre thickness loss in a monkey model of chronic ocular hypertension.

Chronic ocular hypertension (COHT) monkey models were established by destroying the trabecular meshwork, for investigating the relationship between intraocular pressure (IOP) and retinal nerve fibre layer (RNFL) thickness loss. IOP and RNFL thickness were measured before laser injury and weekly thereafter for 27 weeks using Tono Vet and Stratus optical coherence tomography (OCT). The quantitative relationship was as follows: (1) at 32-47 mmHg, the average damage rate was -3.08 ± 0.28 µm/week; (2) at 25-30 mmHg, it was -1.45 ± 0.19 µm/week. The inferior RNFL and superior RNFL turned out to be the most IOP-sensitive quadrants with the rate of RNFL change almost in parallel with IOP levels. The superior sector seemed to be resistant to high IOP conditions until a RNFL loss of ~20 µm was detected in the inferior sector. The rate of RNFL thickness loss was slowed with obvious turning points at RNFL thicknesses of ~75 µm, 65 µm, and 50 µm. The experimental results have achieved research significance. The COHT Monkey was an ideal animal model that can be used for evaluating the relationship between IOP and RNFL damage. Higher IOP was associated with faster RNFL thickness loss. The level of IOP was a vital factor for RNFL damage rate, and baseline/residual RNFL thickness was also important for subsequent RNFL damage. OCT was suitable for measuring RNFL thickness change in COHT monkey models.

BAM15 attenuates transportation-induced apoptosis in iPS-differentiated retinal tissue.

BACKGROUND: BAM15 is a novel mitochondrial protonophore uncoupler capable of protecting mammals from acute renal ischemic-reperfusion injury and cold-induced microtubule damage. The purpose of our study was to investigate the effect of BAM15 on apoptosis during 5-day transportation of human-induced pluripotent stem (hiPS)-differentiated retinal tissue. METHODS: Retinal tissues of 30 days and 60 days were transported with or without BAM15 for 5 days in the laboratory or by real express. Immunofluorescence staining of apoptosis marker cleaved caspase3, proliferation marker Ki67, and neural axon marker NEFL was performed. And expression of apoptotic-related factors p53, NFkappaB, and TNF-a was detected by real-time PCR. Also, location of ganglion cells, photoreceptor cells, amacrine cells, and precursors of neuronal cell types in retinal tissue was stained by immunofluorescence after transportation. Furthermore, cell viability was assessed by CCK8 assay. RESULTS: Results showed transportation remarkably intensified expression of apoptotic factor cleaved caspase3, p53, NFkappaB, and TNF-a, which could be reduced by supplement of BAM15. In addition, neurons were severely injured after transportation, with axons manifesting disrupted and tortuous by staining NEFL. And the addition of BAM15 in transportation was able to protect neuronal structure and increase cell viability without affecting subtypes cells location of retinal tissue. CONCLUSIONS: BAM15 might be used as a protective reagent on apoptosis during transporting retinal tissues, holding great potential in research and clinical applications.

Islet1 and Brn3 Expression Pattern Study in Human Retina and hiPSC-Derived Retinal Organoid.

This study was conducted to determine the dynamic Islet1 and Brn3 (POU4F) expression pattern in the human fetal retina and human-induced pluripotent stem cell- (hiPSC-) derived retinal organoid. Human fetal eyes from 8 to 27 fetal weeks (Fwks), human adult retina, hiPSC-derived retinal organoid from 7 to 31 differentiation weeks (Dwks), and rhesus adult retina were collected for cyrosectioning. Immunofluorescence analysis showed that Islet1 was expressed in retinal ganglion cells in the fetal retina, human adult retina, and retinal organoids. Unexpectedly, after Fwk 20, Brn3 expression gradually decreased in the fetal retina. In the midstage of development, Islet1 was detected in bipolar and developing horizontal cells. As the photoreceptor developed, the Islet1-positive cone precursors gradually became Islet1-negative/S-opsin-positive cones. This study highlights the distinguishing characteristics of Islet1 dynamic expression in human fetal retina development and proposes more concerns which should be taken regarding Brn3 as a cell-identifying marker in mature primate retina.

Short-Term Observation of Ultrasonic Cyclocoagulation in Chinese Patients with End-Stage Refractory Glaucoma: A Retrospective Study.

PURPOSE: To assess the efficacy and safety of HIFU-based ultrasonic cyclocoagulation in Chinese patients with end-stage refractory glaucoma. METHOD: Patients were recruited consecutively from May 2016 to May 2017 in the Zhongshan Ophthalmic Center. Ultrasonic cyclocoagulation was performed on every patient, using the EyeOP1 ultrasound emitting device. Return visits were set at 1 day, 7 days, 1 month, and 3 months after the treatment. An intraocular pressure (IOP) reduction of ≥20% while IOP ≥ 5 mmHg was deemed as success. Mean IOP change was assessed. Efficacy of two modes (6 sectors and 8 sectors) was also compared. Complications were recorded for safety evaluation. RESULTS: 61 eyes were treated in this study. The baseline IOP (mean ± SD) was 41.11 ± 10.65 mmHg. The percentage of IOP reduction after treatment was 29.2%, 43.2%, 34.8%, and 23.1% at 1 day, 7 days, 1 month, and 3 months, respectively. Overall success rate at 3 months was 50.0% (26/52). No significant difference was found between the 6 sectors group and the 8 sectors group in terms of the success rate (48.6% vs. 52.9%, p=0.768) as well as IOP reduction (p=0.417) at 3 months. Primary angle-closure glaucoma (PACG) had the highest success rate (80.0%, 12/15). Scleral thinning existed in 12 eyes, among which 2 developed hypotony (2 mmHg and 3 mmHg). Average pain score decreased massively compared with baseline data. CONCLUSION: With high percentage of IOP reduction and a good safety profile observed in our study, HIFU-based ultrasonic cyclocoagulation might become a promising alternative to cyclodestructive methods. Long-term efficacy and safety need further assessment. The study was registered with Chinese Clinical Trial Registry (http://www.chictr.org.cn; Registration number: ChiCTR-OOC-17014028).

Establishment and Validation of Prognostic Nomograms for Endemic Nasopharyngeal Carcinoma.

BACKGROUND: This study aimed to establish an effective prognostic nomogram with or without plasma Epstein-Barr virus DNA (EBV DNA) for nondisseminated nasopharyngeal carcinoma (NPC). METHODS: The nomogram was based on a retrospective study of 4630 patients who underwent radiotherapy with or without chemotherapy at Sun Yat-sen University Cancer Center from 2007 to 2009. The predictive accuracy and discriminative ability of the nomogram were determined by a concordance index (C-index) and calibration curve and were compared with EBV DNA and the current staging system. The results were validated using bootstrap resampling and a prospective cohort study on 1819 patients consecutively enrolled from 2011 to 2012 at the same institution. All statistical tests were two-sided. RESULTS: Independent factors derived from multivariable analysis of the primary cohort to predict recurrence were age, sex, body mass index (BMI), T stage, N stage, plasma EBV DNA, pretreatment high sensitivity C-reactive protein (hs-CRP), lactate dehydrogenase (LDH), and hemoglobin level (HGB), which were all assembled into the nomogram with (nomogram B) or without EBV DNA (nomogram A). The calibration curve for the probability of recurrence showed that the nomogram-based predictions were in good agreement with actual observations. The C-index of nomogram B for predicting recurrence was 0.728 (P < .001), which was statistically higher than the C-index values for nomogram A (0.690), EBV DNA (0.680), and the current staging system (0.609). The C-index of nomogram B (0.730) and nomogram A (0.681) remained higher for predicting recurrence among patients treated with intensity-modulated radiotherapy (P < .001). The results were confirmed in the validation cohort. CONCLUSIONS: The proposed nomogram with or without plasma EBV DNA resulted in more accurate prognostic prediction for NPC patients.

High-sensitivity C-reactive protein complements plasma Epstein-Barr virus deoxyribonucleic acid prognostication in nasopharyngeal carcinoma: a large-scale retrospective and prospective cohort study.

PURPOSE: To evaluate the effects of combining the assessment of circulating high-sensitivity C-reactive protein (hs-CRP) with that of Epstein-Barr virus DNA (EBV DNA) in the pretherapy prognostication of nasopharyngeal carcinoma (NPC). PATIENTS AND METHODS: Three independent cohorts of NPC patients (training set of n=3113, internal validation set of n=1556, and prospective validation set of n=1668) were studied. Determinants of disease-free survival, distant metastasis-free survival, and overall survival were assessed by multivariate analysis. Hazard ratios and survival probabilities of the patient groups, segregated by clinical stage (T1-2N0-1M0, T3-4N0-1M0, T1-2N2-3M0, and T3-4N2-3M0) and EBV DNA load (low or high) alone, and also according to hs-CRP level (low or high), were compared. RESULTS: Elevated hs-CRP and EBV DNA levels were significantly correlated with poor disease-free survival, distant metastasis-free survival, and overall survival in both the training and validation sets. Associations were similar and remained significant after excluding patients with cardiovascular disease, diabetes, and chronic hepatitis B. Patients with advanced-stage disease were segregated by high EBV DNA levels and high hs-CRP level into a poorest-risk group, and participants with either high EBV DNA but low hs-CRP level or high hs-CRP but low EBV DNA values had poorer survival compared with the bottom values for both biomarkers. These findings demonstrate a significant improvement in the prognostic ability of conventional advanced NPC staging. CONCLUSION: Baseline plasma EBV DNA and serum hs-CRP levels were significantly correlated with survival in NPC patients. The combined interpretation of EBV DNA with hs-CRP levels led to refinement of the risks for the patient subsets, with improved risk discrimination in patients with advanced-stage disease.