Rb deficiency induces p21cip1 expression and delays retinal degeneration in rd1 mice.

Retinitis pigmentosa (RP) is a major cause of inherited blindness, and there is presently no cure for RP. Rd1 mouse is the most commonly used RP animal model. Re-expression of cell cycle proteins in post-mitotic neurons is considered an important mechanism of neurodegenerative diseases, including RP. The retinoblastoma tumor suppressor (Rb) is a major regulator of cell cycle progression, yet its role in rd1 mouse retina and related signaling pathways have never been analyzed. By crossing α-Cre, Rbf/f mice with rd1 mice, p21cip1-/- mice, Cdk1f/f mice and Cdk2f/f mice, we established multiple rd1 mouse models with deletions of Rb gene, Cdkn1a (p21cip1) gene, Cdk1 and Cdk2 gene in the retina. Cdk inhibitor CR8 was injected into the vitreous of rd1 mouse to investigate its effects on photoreceptor survival. Rb gene knockout (KO) induces cell death in excitatory retinal neurons (rods, rod bipolar and ganglions) and ectopic proliferation of retinal cells; but it paradoxically delays the rod death of rd1 mice, which is primarily mediated by the Cdk inhibitor Cdkn1a (p21cip1). Interestingly, p21cip1 protects the ectopic dividing rd1 rod cells by inhibiting Cdk1 and Cdk2. However, inhibiting Cdk1 and Cdk2 in rd1 mice with non-dividing rods only has limited and transient protective effects. Our data suggest that there is no ectopic division of rd1 rod cells, and RbKO induces ectopic division but delays the death of rd1 rod cells. This reveals the important protective role of Rb-p21cip1-Cdk axis in rd1 rod cells. P21cip1 is a potential target for future therapy of RP.

Multimodal imaging and electroretinography highlights the role of VEGF in the laser-induced subretinal fibrosis of monkey.

Age-related macular degeneration (AMD) is a leading cause of blindness. Laser-induced nonhuman primate choroidal neovascularization (CNV) is a widely used animal model of neovascular AMD. Subretinal fibrosis (SFb) is the major limiting factor of effective anti-VEGF therapy for neovascular AMD, yet SFb has never been systematically analyzed in the primate CNV model and if VEGF directly affect SFb is unknown. We recruited a large cohort of rhesus macaques to study the occurrence, multimodal imaging and electroretinography (ERG) features, and related cytokines of SFb. Here we show that among 33 rhesus macaques, 88% CNV eyes developed SFb. Spectral domain optical coherence tomography (SD-OCT) identified four types of subretinal hyper-reflective material (SHRM) of SFb in primate. Multimodal imaging is reliable for monitoring SFb and matches the histological results well. Reduced amplitude of oscillatory potentials correlates with the thinning of inner retina layers and is a possible SFb indicator. Iba1+ microglia/macrophage cells infiltrated in the fibrotic lesions, and aqueous cytokine analysis identified four fibrosis-related factors (GM-CSF, IL-10, TGFß2 and VEGF). Unexpectedly, we found sustained expression of VEGF may be an important inducer of SFb, and anti-VEGF therapy actually partially suppresses SFb. Taken together, our data suggest the laser-induced primate SFb model, coupled with multimodal imaging and ERG recording, is a useful system to dissect the pathogenesis and explore the rationale of treatment for SFb; and combined therapy with anti-VEGF and anti-fibrosis agents is necessary for AMD treatment.

The Role of Gut Microbiome in Autoimmune Uveitis.

The gut microbiome has important physiological functions and plays an indispensable role in the human body. Currently, there are an increasing number of studies revealing the close correlation between dysbiosis of the gut microbiome and a variety of autoimmune diseases, including autoimmune uveitis. This brief review summarizes recent literature regarding the relationship between dysbiosis and the occurrence and development of autoimmune uveitis. Dysbiosis participates in the pathogenesis of autoimmune uveitis largely by 4 mechanisms: antigenic mimicry, disturbance of intestinal immune homeostasis, destruction of the intestinal barrier, and reduction of beneficial anti-inflammatory metabolites. Further elucidation of these mechanisms will facilitate the treatment of the gut-microbiome-relevant autoimmune diseases by potential therapeutic strategies, such as antibiotics, probiotics, diet modifications, and fecal microbial transplantation.

Performance verification of a biochemical detection system for hydrothorax and ascites and clinical diagnostic accuracy evaluation of exudate and tuberculous effusion.

Background: Lactate dehydrogenase (LDH), total protein (TP) and glucose (Glu) in pleural hydrothorax and ascites can be used in the diagnosis of exudate, and adenosine deaminase (ADA) can be used in the diagnosis of tuberculous effusion. However, the manufacturers do not claim that their biochemical reagents can be used to detect hydrothorax and ascites samples. Therefore, medical laboratories must conduct suitability studies on biochemical reagents for hydrothorax and ascites samples to comply with regulatory requirements for humor detection. This study aimed to verify the analytical performance and clinical diagnostic accuracy of the Mindray biochemical reagents, including LDH, TP, Glu and ADA, for hydrothorax and ascites. Methods: The repeatability, detection limits and reference intervals of Mindray biochemical reagents (LDH, TP, Glu, ADA) in detecting hydrothorax and ascites were determined. The comparison of different measurement procedures was performed. Meanwhile, the diagnostic accuracy of LDH, TP, Glu and ADA were assessed. Results: The quality control results of LDH, TP, Glu, and ADA were all under control. The repeatability coefficient of variation (%) of LDH, TP, Glu, and ADA were all less than 1%. The limits of blank of LDH, TP, Glu, and ADA were 0.33 U/L, 0.45 g/L, 0.00 mmol/L, and 0.04 U/L, respectively; the limits of detection were 1.57 U/L, 1.85 g/L, 0.05 mmol/L, and 0.12 U/L, respectively. Compared with the reference measurement program, the correlation coefficients of LDH, TP, Glu and ADA were 0.9931, 0.9983, 0.9996 and 0.9966, respectively; the regression equations were y=1.0082x-10.06, y=0.9965x-0.4732, y=0.9903x+0.0522 and y=1.0051x-0.0232, respectively. The reference intervals of LDH, TP, Glu, and ADA in hydrothorax and ascites were ≤198.39 U/L, ≤32.97 g/L, ≥5.03 mmol/L. and ≤11.00 U/L respectively. For differentiating between exudates and transudates, the area under the curve (AUC) of LDH, TP, and Glu were 0.913, 0.875, and 0.767, respectively; the AUC of ADA for the differential diagnosis of tuberculous and nontuberculous effusions was 0.876. Conclusions: The LDH, TP, Glu, and ADA assays were validated for use with the Mindray BS-2800 analyzer for hydrothorax and ascites evaluation. LDH, TP, and Glu in hydrothorax and ascites are applicable to the differential diagnosis of exudates and transudates; ADA in hydrothorax and ascites can be employed to differentiate and diagnose tuberculous and nontuberculous effusions.

Sub-lobar resection versus lobectomy for challenging intraoperative frozen sections in lung adenocarcinoma within 3 cm.

OBJECTIVES: Intraoperative frozen section (FS) analysis is pivotal in guiding surgical interventions for early-stage lung adenocarcinoma. However, the challenge arises when distinguishing between Minimally Invasive Adenocarcinoma (MIA) and Invasive Adenocarcinoma (IAC) poses diagnostic difficulties. This study investigates the prognosis and clinicopathological characteristics of patients encountering this diagnostic challenge. METHODS: We conducted a retrospective analysis of 7082 intraoperative FSs from early-stage lung adenocarcinoma cases. The cases with pulmonary nodules within 3 cm and diagnosed as indeterminate FSs were included. We analyzed baseline data, computed tomography (CT) findings, and pathological characteristics. Prognostic data were obtained from patients with confirmed IAC diagnoses through final pathological examination. RESULTS: Out of 7082 FSs, 551 cases presented challenges in distinguishing between MIA and IAC. Upon final pathological examination, 233 cases were identified as IAC, while 314 were classified as MIA. The median invasive pathological size in the IAC group was larger than that in the MIA group (0.6 cm vs 0.3 cm). 131 cases (56.2 %) with IAC underwent lobectomy, while 102 cases (43.8 %) underwent sub-lobar resection. Among the MIA cases, 220 cases (69.8 %) underwent sub-lobar resection, while 95 cases (30.2 %) underwent lobectomy. No recurrence and disease specific death was observed during the follow-up period, regardless of surgical strategy. CONCLUSIONS: Indeterminate intraoperative FSs, posing diagnostic challenges in distinguishing between MIA and IAC. Sub-lobar resection presented the same long term survival benefit compared with the lobectomy for indeterminate lung adenocarcinoma within 3 cm during intraoperative FSs.

Utilization of intraoperative indocyanine green fluorescence imaging to identify vascular anatomy in severe pleural adhesions in uniportal video-assisted thoracoscopic surgery: a case report.

Background: Extensive and dense pleural adhesion is a serious challenge in video-assisted thoracoscopic surgery (VATS), in which identification of vessels and their anatomical spaces is difficult. Once critical vessel is damaged while dissecting adhesion in VATS, leading to fatal hemorrhage, the surgeon will have to switch to thoracotomy. This is the first report of a case in which intraoperative indocyanine green (ICG) fluorescence imaging was used to identify critical vessels in severe pleural adhesions in uniportal VATS. Case Description: The patient (67-year-old male) with an 8-year history of tuberculosis and severe mixed ventilation dysfunction underwent a standardized wedge resection due to chest computed tomography (CT) scan that revealed a 2.6-cm nodule in the right upper lung. Intraoperatively, the superior vena cava and azygos vein were successfully identified and safely dissected using ICG fluorescence imaging in the presence of extensive and dense pleural adhesions. The chest drainage tube was removed on postoperative day (POD) 3, and patient was released from hospital on POD 5. The patient recovered well and no complication was observed in the follow-up. Conclusions: The ICG fluorescence imaging is used to illustrate the vessels and help to dissect them safely, which is a feasible, visualizable, and user-friendly method in severe pleural adhesions in uniportal VATS.

Hyperlipidemia and lipid-lowering therapy in diabetic retinopathy (DR): A bibliometric study and visualization analysis in 1993-2023.

Background: Diabetic retinopathy (DR) is a common complication in diabetic patients. DR is also a neurodegenerative disease. Patients with hyperglycemia, hyperlipidemia, and hypertension are vulnerable to retinopathy development. While the roles of blood glucose and blood pressure in the development of retinopathy have been extensively studied, the relationship between body fat and DR pathogenesis and the impact of lipid-reducing drugs on DR has just emerged as a research hotspot in DR study. We aim to visualize the contributions and cooperation of reporters, organizations, and nations, in addition to the research hotspots and trends in DR-related lipid research from 1993 to 2023, by bibliometric analysis. Methods: We extracted all publications about DR-related lipid research from 1993 to 2023 from the Web of Science Core Collection, and bibliometric features were studied using VOSviewer and the CiteSpace program. Results: 1402 documents were retrieved. The number of studies has risen consistently for three decades, from an average of 16.8/year in the 1990s to 28.8/year in the 2000s, 64.5/year in 2010s, and reached 112/year in 2020-2022, confirming they are hot research topic in the field. These reports were from 93 nations/regions, with the USA, China, Japan, Australia, and England taking the leading positions. Diabetes Research and Clinical Practice was the journal that published the most studies, and Diabetes Care was the most quoted. We identified 6979 authors, with Wong TY having the most papers and being the most commonly co-cited. The most popular keyword, according to our research, is diabetic retinopathy. Oxidative stress, diabetic macular edema (DME), lipid peroxidation, and other topics have often been investigated. Conclusion: DR-related lipid research is conducted mainly in North America, Asia, Oceania, and Europe. Much study has centered on the relationship between lipid-lowering therapy and DR pathogenesis. These studies strongly support using lipid-reducing medications (fenofibrate, statins, and omega-3 PUFAs), combined with hyperglycemia and hypertension therapy, to prevent and treat DR. However, the impact of fenofibrate or statin on retinopathy is not correlated with their action on blood lipid profiles. Thus, more randomized clinical trials with primary endpoints related to DR in T1D or T2D are merited. In addition, the lipid biomarker for DR (lipid aldehydes, ALEs, and cholesterol crystals), the action of lipid-reducing medicines on retinopathy, the mechanism of lipid-lowering medications preventing or curing DR, and ocular delivery of lipid-lowering drugs to diabetic patients are predicted as the research focus in the future in the DR-related lipid research field.

The role of the microbiota in glaucoma.

Glaucoma is a common irreversible vision loss disorder because of the gradual loss of retinal ganglion cells (RGCs) and the optic nerve axons. Major risk factors include elder age and high intraocular pressure (IOP). However, high IOP is neither necessary nor sufficient to cause glaucoma. Some non-IOP signaling cascades can mediate RGC degeneration. In addition, gender, diet, obesity, depression, or anxiety also contribute to the development of glaucoma. Understanding the mechanism of glaucoma development is crucial for timely diagnosis and establishing new strategies to improve current IOP-reducing therapies. The microbiota exerts a marked influence on the human body during homeostasis and disease. Many glaucoma patients have abnormal compositions of the microbiota (dysbiosis) in multiple locations, including the ocular surface, intraocular cavity, oral cavity, stomach, and gut. Here, we discuss findings in the last ten years or more about the microbiota and metabolite changes in animal models, patients with three risk factors (aging, obesity, and depression), and glaucoma patients. Antigenic mimicry and heat stress protein (HSP)-specific T-cell infiltration in the retina may be responsible for commensal microbes contributing to glaucomatous RGC damage. LPS-TLR4 pathway may be the primary mechanism of oral and ocular surface dysbiosis affecting glaucoma. Microbe-derived metabolites may also affect glaucoma pathogenesis. Homocysteine accumulation, inflammatory factor release, and direct dissemination may link gastric H. pylori infection and anterior chamber viral infection (such as cytomegalovirus) to glaucoma. Potential therapeutic protocols targeting microbiota include antibiotics, modified diet, and stool transplant. Later investigations will uncover the underlying molecular mechanism connecting dysbiosis to glaucoma and its clinical applications in glaucoma management.

The heterogeneity of astrocytes in glaucoma.

Glaucoma is a leading cause of blindness with progressive degeneration of retinal ganglion cells. Aging and increased intraocular pressure (IOP) are major risk factors. Lowering IOP does not always stop the disease progression. Alternative ways of protecting the optic nerve are intensively studied in glaucoma. Astrocytes are macroglia residing in the retina, optic nerve head (ONH), and visual brain, which keep neuronal homeostasis, regulate neuronal activities and are part of the immune responses to the retina and brain insults. In this brief review, we discuss the activation and heterogeneity of astrocytes in the retina, optic nerve head, and visual brain of glaucoma patients and animal models. We also discuss some recent transgenic and gene knockout studies using glaucoma mouse models to clarify the role of astrocytes in the pathogenesis of glaucoma. Astrocytes are heterogeneous and play crucial roles in the pathogenesis of glaucoma, especially in the process of neuroinflammation and mitochondrial dysfunction. In astrocytes, overexpression of Stat3 or knockdown of IκKß/p65, caspase-8, and mitochondrial uncoupling proteins (Ucp2) can reduce ganglion cell loss in glaucoma mouse models. Based on these studies, therapeutic strategies targeting the heterogeneity of reactive astrocytes by enhancing their beneficial reactivity or suppressing their detrimental reactivity are alternative options for glaucoma treatment in the future.

The Role of the Microbiota in Graves' Disease and Graves' Orbitopathy.

Graves' disease (GD) is a clinical syndrome with an enlarged and overactive thyroid gland, an accelerated heart rate, Graves' orbitopathy (GO), and pretibial myxedema (PTM). GO is the most common extrathyroidal complication of GD. GD/GO has a significant negative impact on the quality of life. GD is the most common systemic autoimmune disorder, mediated by autoantibodies to the thyroid-stimulating hormone receptor (TSHR). It is generally accepted that GD/GO results from complex interactions between genetic and environmental factors that lead to the loss of immune tolerance to thyroid antigens. However, the exact mechanism is still elusive. Systematic investigations into GD/GO animal models and clinical patients have provided important new insight into these disorders during the past 4 years. These studies suggested that gut microbiota may play an essential role in the pathogenesis of GD/GO. Antibiotic vancomycin can reduce disease severity, but fecal material transfer (FMT) from GD/GO patients exaggerates the disease in GD/GO mouse models. There are significant differences in microbiota composition between GD/GO patients and healthy controls. Lactobacillus, Prevotella, and Veillonella often increase in GD patients. The commonly used therapeutic agents for GD/GO can also affect the gut microbiota. Antigenic mimicry and the imbalance of T helper 17 cells (Th17)/regulatory T cells (Tregs) are the primary mechanisms proposed for dysbiosis in GD/GO. Interventions including antibiotics, probiotics, and diet modification that modulate the gut microbiota have been actively investigated in preclinical models and, to some extent, in clinical settings, such as probiotics (Bifidobacterium longum) and selenium supplements. Future studies will reveal molecular pathways linking gut and thyroid functions and how they impact orbital autoimmunity. Microbiota-targeting therapeutics will likely be an essential strategy in managing GD/GO in the coming years.

Clinical Pathological Features and Current Animal Models of Type 3 Macular Neovascularization.

Type 3 macular neovascularization (MNV3), or retinal angiomatous proliferation (RAP), is a distinct type of neovascular age-related macular degeneration (AMD), which is a leading cause of vision loss in older persons. During the past decade, systematic investigation into the clinical, multimodal imaging, and histopathological features and therapeutic outcomes has provided important new insight into this disease. These studies favor the retinal origin of MNV3 and suggest the involvement of retinal hypoxia, inflammation, von Hippel-Lindau (VHL)-hypoxia-inducible factor (HIF)-vascular endothelial growth factor (VEGF) pathway, and multiple cell types in the development and progression of MNV3. Several mouse models, including the recently built Rb/p107/Vhl triple knockout mouse model by our group, have induced many of the histological features of MNV3 and provided much insight into the underlying pathological mechanisms. These models have revealed the roles of retinal hypoxia, inflammation, lipid metabolism, VHL/HIF pathway, and retinoblastoma tumor suppressor (Rb)-E2F cell cycle pathway in the development of MNV3. This article will summarize the clinical, multimodal imaging, and pathological features of MNV3 and the diversity of animal models that exist for MNV3, as well as their strengths and limitations.

Body Composition Parameters May Be Prognostic Factors in Upper Urinary Tract Urothelial Carcinoma Treated by Radical Nephroureterectomy.

OBJECTIVE: This study assessed the association between body composition and prognosis of patients with upper urinary tract urothelial carcinoma (UTUC) patients treated by radical nephroureterectomy. METHODS: We retrospectively collected baseline data on age, sex, body mass index (BMI), hypertension, diabetes, and tumor-related factors. Computed tomography (CT) scans were performed to measure body composition parameters such as muscle attenuation (MA), total abdominal muscle area (TAMA), visceral fat area (VFA), intermuscular fat area (IMF), and lateral/posterior perirenal fat thickness (L/P PNF), visceral fat density (VD), and subcutaneous fat density (SD). Patient follow-up was conducted via telephone or in the clinic. The endpoints of follow-up were all-cause death, local progression or distant metastasis. Survival analysis was analyzed using the Kaplan-Meier method, and risk factors associated with prognosis were identified using univariate and multivariate Cox proportional hazard analyses. RESULTS: Among the 273 UTUC patients (median age, 68 years) enrolled in our study, 102 had a BMI > 24.0, 100 suffered from diabetes, and 120 had hypertension. A large proportion of patients (189) had high grade tumors. Across all patients, 1- and 3-year rates for overall survival were 86.45% and 75.55%; local progression-free survival, 92.11% and 89.67%; and distant metastasis-free survival, 85.23% and 80.17%. Based on the Cox regression analysis, MA, IMF, TAMA, TPA, TPT, APT, SMI and PMI significantly reduced the risk of local progression (p < 0.05), while PPNF = 1 point reduced the risk of distant metastasis (p < 0.05). Overall survival was significantly associated with MA, TAMA, and SMI (p < 0.05). CONCLUSION: Our findings illustrate that body composition parameters can act as independent predictors of prognosis in UTUC patients who underwent RNU. These results can help improve stratification of patients and optimize postoperative treatment.

Interaction Between Mesenchymal Stem Cells and Retinal Degenerative Microenvironment.

Retinal degenerative diseases (RDDs) are a group of diseases contributing to irreversible vision loss with yet limited therapies. Stem cell-based therapy is a promising novel therapeutic approach in RDD treatment. Mesenchymal stromal/stem cells (MSCs) have emerged as a leading cell source due to their neurotrophic and immunomodulatory capabilities, limited ethical concerns, and low risk of tumor formation. Several pre-clinical studies have shown that MSCs have the potential to delay retinal degeneration, and recent clinical trials have demonstrated promising safety profiles for the application of MSCs in retinal disease. However, some of the clinical-stage MSC therapies have been unable to meet primary efficacy end points, and severe side effects were reported in some retinal "stem cell" clinics. In this review, we provide an update of the interaction between MSCs and the RDD microenvironment and discuss how to balance the therapeutic potential and safety concerns of MSCs' ocular application.

Rb1/Rbl1/Vhl loss induces mouse subretinal angiomatous proliferation and hemangioblastoma.

Von Hippel-Lindau (Vhl) protein inhibits hypoxia-inducible factor (Hif), yet its deletion in murine retina does not cause the extensive angiogenesis expected with Hif induction. The mechanism is unclear. Here we show that retinoblastoma tumor suppressor (Rb1) constrains expression of Hif target genes in the Vhl-/- retina. Deleting Rb1 induced extensive retinal neovascularization and autophagic ablation of photoreceptors in the Vhl-/- retina. RNA-sequencing, ChIP, and reporter assays showed Rb1 recruitment to and repression of certain Hif target genes. Activating Rb1 by deleting cyclin D1 induced a partial defect in the retinal superficial vascular plexus. Unexpectedly, removing Vhl suppressed retinoblastoma formation in murine Rb1/Rbl1-deficient retina but generated subretinal vascular growths resembling retinal angiomatous proliferation (RAP) and retinal capillary hemangioblastoma (RCH). Most stromal cells in the RAP/RCH-like lesions were Sox9+, suggesting a Müller glia origin, and expressed Lgals3, a marker of human brain hemangioblastoma. Thus, the Rb family limit Hif target gene expression in the Vhl-/- retina, and removing this inhibitory signal generates new models for RAP and RCH.

Rb is required for retinal angiogenesis and lamination.

Retinoblastoma tumor suppressor (Rb) promotes cell cycle exit, survival, differentiation, and tumor suppression in the retina. Here, we show it is also essential for vascularization and lamination. Despite minimal effects on Hif1a target expression, intraretinal vascular plexi did not form in the Rb -/- murine retina. Deleting adenovirus E2 promoter binding factor 3 (E2f3), which rescues starburst amacrine cell differentiation, or E2f2, had no effect, but deleting E2f1, which promotes neuronal cell cycle exit and survival, restored retinal vasculature. We specifically linked cell loss to the defect because removing Bax rescued rod and bipolar neurons and the vasculature, but not cell cycle exit. Despite rescuing Rb -/- neurons, Bax deletion exacerbated a delay in outer retina lamination, and exposed a requirement for Rb in inner retina lamination. The latter resembled Sem5 or FAT atypical cadherin 3 (Fat3) mutants, but expression of Sem5/Fat3 pathway components, or that of Neogenin, which perturbs migration in the Rb -/- cortex, was unchanged. Instead, lamination defects correlated with ectopic division, and were E2f1-dependent, implicating the cell cycle machinery. These in vivo studies expose new developmental roles for Rb, pinpoint aberrant E2f1 and Bax activity in neuronal death and vascular loss, and further implicate E2f1 in defective lamination. Links between Rb, angiogenesis and lamination have implications for the treatment of neovascularization, neurodegeneration and cancer.

Potential new antitumor agents from an innovative combination of camphorato, a ramification of traditional Chinese medicine, with a platinum moiety.

Eight new camphorato platinum complexes have been synthesized and evaluated for their in vitro cytotoxicity against HL-60 human leukemia, 3AO human ovarian carcinoma, BEL-7402 human hepatocarcinoma, and A549 human lung carcinoma cell lines. Most complexes showed good cytotoxic activity against the above-selected cell lines. Among the complexes, two compounds were assayed for their in vivo antitumor activity against LS-174T human colon carcinoma cells implanted in mice. One complex exhibited not only higher in vivo antitumor activity, but also less toxicity than oxaliplatin when it was administered intravenously at a dose of 6 mg/kg three times.