Cryopreservation in Ophthalmology

Amniotic membranes (AMs) and corneas are critical materials in ocular surface reconstruction. AM has specific structures (e.g., basement and two types of cells with stemness characteristics: amniotic epithelial cells and amniotic mesenchymal cells), which contribute to its attractive physical and biological properties that make it fundamental to clinical application. The corneal endothelial cell is a vital part of the cornea, which can influence postoperative vision directly. However, widespread use of fresh AM and cornea has been limited due to their short use span and safety concerns. To overcome these concerns, different preservation methods have been introduced. Cryopreservation is distinguished from many preservation methods for its attractive advantages of prolonged use span, optimally retained tissue structure, and minimized infection risk. This review will focus on recent advances of cryopreserved AM and cornea, including different cryopreservation methods and their indications in ophthalmology

PO-099 Effects of long-term high-intensity exercise training on renal local renin-angiotensin system in rats

Objective Exercise stimulation to the body of the first stress: blood changes, regulated by the renin-angiotensin system. Long-term high-intensity exercise training will lead to changes in kidney structure and function, resulting in renal injury. This will not only affect training and competition, but also affect the health of athletes, thus it is becoming an influential factor in the occurrence and development of excessive fatigue. At present, the mechanism of renal injury and proteinuria caused by exercise is not very clear. Therefore, this study conducted an in-depth study on the upstream mechanism of renal blood flow changes from the molecular level and explored the effects of 6-week high-intensity exercise training on the renal local renin-angiotensin system in rats. Methods 30 SD male rats (8 weeks old) were randomly divided into control group (10) and exercise group (20). Then according to different time, the exercise group (20) was divided into two groups on average, which are 0h group and 24h group. The rats in the control group did not do any exercise, and the rats in the exercise group were trained to run on the platform with high intensity for 6 weeks. At the beginning of the sixth week, the urine of rats from different group were selected randomly, and the urine NGAL, urinary microalbumin (mAlb), urinary creatinine (UCr) and total urine protein (TP) were tested, to determine the rat model of exercise-induced renal injury with proteinuria. After training on the 6th weekend, the renal tissue renin activity and angiotensin Ⅱ (Ang Ⅱ) content in right kidney were tested. At the same time, the left kidney of the rat was taken to make HE staining sections. Results (1) The mAlb, Alb/Cr and TP of the exercise group were higher than those of the control group, while the UCr content was lower than that of the control group. There was a significant difference between mAlb and TP in the exercise group and the control group(p<0.05). UCr, mAlb/Cr were significantly different from the control group(p<0.01). The urine NGAL concentration in the exercise group was higher than that in the control group, and the difference was significant(p<0.05). (2) The glomerulus of the rats in the 0h group showed obvious congestion, swelling and erythrocyte exudation. The tissue morphology of rats in the 24h group recovered slightly, but it was still different from the control group. (3) The renin activity and Ang Ⅱ were the lowest in the 0h group, and the renin activity and Ang Ⅱ were highest in the 24h group. There is a very significant difference between the groups(p<0.01). Conclusions (1) The 6-week high-intensity training used in this study increased the levels of NGAL, TP, and mAlb in the urine of rats, and successfully established a rat model of exercise-induced renal injury with proteinuria. (2) Long-term high-intensity exercise training can cause obvious congestion, swelling, erythrocyte exudation in rat glomeruli, which can’t return to the quiet level 24 hours after exercise. (3) Renal injury caused by exercise decreased the expression of local renin activity and angiotensin II in the kidney, and the recovery of renal renin activity and angiotensin II was increased 24h after exercise

By integrating single-cell RNA-seq and bulk RNA-seq in sphingolipid metabolism, CACYBP was identified as a potential therapeutic target in lung adenocarcinoma

BackgroundLung adenocarcinoma (LUAD) is a heterogeneous disease with a dismal prognosis for advanced tumors. Immune-associated cells in the microenvironment substantially impact LUAD formation and progression, which has gained increased attention in recent decades. Sphingolipids have a profound impact on tumor formation and immune infiltration. However, few researchers have focused on the utilization of sphingolipid variables in the prediction of LUAD prognosis. The goal of this work was to identify the major sphingolipid-related genes (SRGs) in LUAD and develop a valid prognostic model based on SRGs.MethodsThe most significant genes for sphingolipid metabolism (SM) were identified using the AUCell and WGCNA algorithms in conjunction with single-cell and bulk RNA-seq. LASSO and COX regression analysis was used to develop risk models, and patients were divided into high-and low-risk categories. External nine provided cohorts evaluated the correctness of the models. Differences in immune infiltration, mutation landscape, pathway enrichment, immune checkpoint expression, and immunotherapy were also further investigated in distinct subgroups. Finally, cell function assay was used to verify the role of CACYBP in LUAD cells.ResultsA total of 334 genes were selected as being most linked with SM activity for further investigation, and a risk model consisting of 11 genes was established using lasso and cox regression. According to the median risk value, patients were split into high- and low-risk groups, and the high-risk group had a worse prognosis. The low-risk group had more immune cell infiltration and higher expression of immune checkpoints, which illustrated that the low-risk group was more likely to benefit from immunotherapy. It was verified that CACYBP could increase the ability of LUAD cells to proliferate, invade, and migrate.ConclusionThe eleven-gene signature identified in this research may help physicians create individualized care plans for LUAD patients. CACYBP may be a new therapeutic target for patients with advanced LUAD

Effects of hypersplenism on the outcome of hepatectomy in hepatocellular carcinoma with hepatitis B virus related portal hypertension

BackgroundAlthough hepatectomy plus splenectomy is not regularly recommended for hepatocellular carcinoma (HCC) with portal hypertension related hypersplenism due to the high risk accompanied with surgical procedures for now. Many researchers still believe that hypersplenism is a controversial adverse prognostic factor for HCC patients. Thus, the primary objective of the study was to determine the effects of hypersplenism on the prognosis of these patients during and after hepatectomy.MethodsA total of 335 patients with HBV-related HCC who underwent surgical resection as primary intervention were included in this study and categorized into three groups. Group A consisted of 226 patients without hypersplenism, Group B included 77 patients with mild hypersplenism, and Group C contained 32 patients with severe hypersplenism. The influence of hypersplenism on the outcome during the perioperative and long-term follow-up periods was analyzed. The independent factors were identified using the Cox proportional hazards regression model.ResultsThe presence of hypersplenism is associated with longer hospital stays, more postoperative blood transfusions, and higher complication rates. The overall survival (OS, P = 0.020) and disease-free survival (DFS, P = 0.005) were significantly decreased in Group B compared to those in Group A. Additionally, the OS (P = 0.014) and DFS (P = 0.005) were reduced in Group C compared to those in Group B. Severe hypersplenism was a significant independent prognostic variable for both OS and DFS.ConclusionSevere hypersplenism prolonged the hospital stay, increased the rate of postoperative blood transfusion, and elevated the incidence of complications. Furthermore, hypersplenism predicted lower overall and disease-free survivals

Cuproptosis-related lncRNA signatures: Predicting prognosis and evaluating the tumor immune microenvironment in lung adenocarcinoma

BackgroundCuproptosis, a unique kind of cell death, has implications for cancer therapy, particularly lung adenocarcinoma (LUAD). Long non-coding RNAs (lncRNAs) have been demonstrated to influence cancer cell activity by binding to a wide variety of targets, including DNA, RNA, and proteins.MethodsCuproptosis-related lncRNAs (CRlncRNAs) were utilized to build a risk model that classified patients into high-and low-risk groups. Based on the CRlncRNAs in the model, Consensus clustering analysis was used to classify LUAD patients into different subtypes. Next, we explored the differences in overall survival (OS), the tumor immune microenvironment (TIME), and the mutation landscape between different risk groups and molecular subtypes. Finally, the functions of LINC00592 were verified through in vitro experiments.ResultsPatients in various risk categories and molecular subtypes showed statistically significant variations in terms of OS, immune cell infiltration, pathway activity, and mutation patterns. Cell experiments revealed that LINC00592 knockdown significantly reduced LUAD cell proliferation, invasion, and migration ability.ConclusionThe development of a trustworthy prediction model based on CRlncRNAs may significantly aid in the assessment of patient prognosis, molecular features, and therapeutic modalities and may eventually be used in clinical applications

Protection of Human Umbilical Vein Endothelial Cells against Oxidative Stress by MicroRNA-210

Oxidative stress induces endothelial cell apoptosis and promotes atherosclerosis development. MicroRNA-210 (miR-210) is linked with apoptosis in different cell types. This study aimed to investigate the role of miR-210 in human umbilical vein endothelial cells (HUVECs) under oxidative stress and to determine the underlying mechanism. HUVECs were treated with different concentrations of hydrogen peroxide (H2O2), and cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and ATP assay. To evaluate the role of miR-210 in H2O2-mediated apoptosis, gain-and-loss-of-function approaches were used, and the effects on apoptosis and reactive oxygen species (ROS) level were assayed using flow cytometry. Moreover, miR-210 expression was detected by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), and expression of the following apoptosis-related genes was assessed by qRT-PCR and Western blot at the RNA and protein level, respectively: caspase-8-associated protein 2 (CASP8AP2), caspase-8, and caspase-3. The results showed that H2O2 induced apoptosis in HUVECs in a dose-dependent manner and increased miR-210 expression. Overexpression of miR-210 inhibited apoptosis and reduced ROS level in HUVECs treated with H2O2. Furthermore, miR-210 downregulated CASP8AP2 and related downstream caspases at protein level. Thus, under oxidative stress, miR-210 has a prosurvival and antiapoptotic effect on HUVECs by reducing ROS generation and downregulating the CASP8AP2 pathway

Serum Levels of FGF-21 Are Increased in Coronary Heart Disease Patients and Are Independently Associated with Adverse Lipid Profile

BACKGROUND: Fibroblast growth factor 21 (FGF-21) is a metabolic regulator with multiple beneficial effects on glucose homeostasis and lipid metabolism in animal models. The relationship between plasma levels of FGF-21 and coronary heart disease (CHD) in unknown. METHODOLOGY/PRINCIPAL FINDINGS: This study aimed to investigate the correlation of serum FGF-21 levels and lipid metabolism in the patients with coronary heart disease. We performed a logistic regression analysis of the relation between serum levels of FGF-21 and CHD patients with and without diabetes and hypertension. This study was conducted in the Departments of Endocrinology and Cardiovascular Diseases at two University Hospitals. Participants consisted of one hundred and thirty-five patients who have been diagnosed to have CHD and sixty-one control subjects. Serum FGF-21 level and levels of fasting blood glucose; triglyceride; apolipoprotein B100; HOMA-IR; insulin; total cholesterol; HDL-cholesterol; LDL-cholesterol; and C-reactive protein were measured. We found that median serum FGF-21 levels were significantly higher in CHD than that of control subjects (P<0.0001). Serum FGF-21 levels in CHD patients with diabetes, hypertension, or both were higher than that of patients without these comorbidities. Serum FGF-21 levels correlated positively with triglycerides, fasting blood glucose, apolipoprotein B100, insulin and HOMA-IR but negatively with HDL-C and apolipoprotein A1 after adjusting for BMI, diabetes and hypertension. Logistic regression analysis demonstrated that FGF-21 showed an independent association with triglyceride and apolipoprotein A1. CONCLUSIONS/SIGNIFICANCE: High levels of FGF-21 are associated with adverse lipid profiles in CHD patients. The paradoxical increase of serum FGF-21 in CHD patients may indicate a compensatory response or resistance to FGF-21

H5N1 avian influenza re-emergence of Lake Qinghai: phylogenetic and antigenic analyses of the newly isolated viruses and roles of migratory birds in virus circulation

Highly pathogenic avian influenza H5N1 virus has swept west across the globe and caused serious debates on the roles of migratory birds in virus circulation since the first large-scale outbreak in migratory birds of Lake Qinghai, 2005. In May 2006, another outbreak struck Lake Qinghai and six novel strains were isolated. To elucidate these QH06 viruses, the six isolates were subjected to whole-genome sequencing. Phylogenetic analyses show that QH06 viruses are derived from the lineages of Lake Qinghai, 2005. Five of the six novel isolates are adjacent to the strain A/Cygnus olor/Croatia/1/05, and the last one is related to the strain A/duck/Novosibirsk/02/05, an isolate of the flyway. Antigenic analyses suggest that QH06 and QH05 viruses are similar to each other. These findings implicate that QH06 viruses of Lake Qinghai may travel back via migratory birds, though not ruling out the possibility of local circulation of viruses of Lake Qinghai