Granzyme B degrades extracellular matrix and promotes inflammation and choroidal neovascularization.

Age-related macular degeneration (AMD) is a common retinal neurodegenerative disease among the elderly. Neovascular AMD (nAMD), a leading cause of AMD-related blindness, involves choroidal neovascularization (CNV), which can be suppressed by anti-angiogenic treatments. However, current CNV treatments do not work in all nAMD patients. Here we investigate a novel target for AMD. Granzyme B (GzmB) is a serine protease that promotes aging, chronic inflammation and vascular permeability through the degradation of the extracellular matrix (ECM) and tight junctions. Extracellular GzmB is increased in retina pigment epithelium (RPE) and mast cells in the choroid of the healthy aging outer retina. It is further increased in donor eyes exhibiting features of nAMD and CNV. Here, we show in RPE-choroidal explant cultures that exogenous GzmB degrades the RPE-choroid ECM, promotes retinal/choroidal inflammation and angiogenesis while diminishing anti-angiogenic factor, thrombospondin-1 (TSP-1). The pharmacological inhibition of either GzmB or mast-cell degranulation significantly reduces choroidal angiogenesis. In line with our in vitro data, GzmB-deficiency reduces the extent of laser-induced CNV lesions and the age-related deterioration of electroretinogram (ERG) responses in mice. These findings suggest that targeting GzmB, a serine protease with no known endogenous inhibitors, may be a potential novel therapeutic approach to suppress CNV in nAMD.

[Advances in non-invasive treatment of aortic diseases].

With the further development and long-term follow-up of endovascular treatment for aortic diseases, increasing evidence shows that in many cases, there are difficulties in the diagnosis of causes, decision-making of treatment timing, and lack of effective evaluation of treatment prognosis in endovascular treatments. Therefore, it is necessary to conduct in-depth research on non-invasive treatment including prevention, diagnosis, treatment, and prediction of aortic diseases. The non-invasive treatment of aortic disease is mainly applied to high-risk populations with aortic dissection, regulating key targets and mechanisms, and adopting drug intervention in advance to achieve the goal of controlling aortic dilation and preventing the occurrence of dissection. It also conducts precise multi omics analysis to determine the optimal intervention timing and treatment strategy, and aims at complications related to aortic disease or endovascular treatment for patients with a positive family history of aortic dilation and those who have developed aortic dissection. Precise regulation can control the progression of aortic aneurysm and aortic dissection, delay or achieve long-term stable coexistence with aortic disease, and even reverse disease progression and achieve benign aortic remodeling through new intervention vectors. Ultimately achieving the ideal state of complete thrombosis and mechanized healing of the aortic aneurysm or aortic dissection false lumen.

Granzyme B Contributes to Choroidal Neovascularization and Age-Related Macular Degeneration Through Proteolysis of Thrombospondin-1.

Age-related macular degeneration (AMD) is a leading cause of irreversible central vision loss in the elderly. The pathology of neovascular age-related macular degeneration (nAMD), also known as wet AMD, is associated with an abnormal blood vessel growth in the eye and involves an imbalance of proangiogenic and antiangiogenic factors. Thrombospondin (TSP)-1 and TSP-2 are endogenous matricellular proteins that inhibit angiogenesis. TSP-1 is significantly diminished in eyes with AMD, although the mechanisms involved in its reduction are unknown. Granzyme B (GzmB) is a serine protease with an increased extracellular activity in the outer retina and choroid of human eyes with nAMD-related choroidal neovascularization (CNV). This study investigated whether TSP-1 and TSP-2 are GzmB substrates using in silico and cell-free cleavage assays and explored the relationship between GzmB and TSP-1 in human eyes with nAMD-related CNV and the effect of GzmB on TSP-1 in retinal pigment epithelial culture and an explant choroid sprouting assay (CSA). In this study, TSP-1 and TSP-2 were identified as GzmB substrates. Cell-free cleavage assays substantiated the GzmB proteolysis of TSP-1 and TSP-2 by showing dose-dependent and time-dependent cleavage products. TSP-1 and TSP-2 proteolysis were hindered by the inhibition of GzmB. In the retinal pigment epithelium and choroid of human eyes with CNV, we observed a significant inverse correlation between TSP-1 and GzmB, as indicated by lower TSP-1 and higher GzmB immunoreactivity. In CSA, the vascular sprouting area increased significantly with GzmB treatment and reduced significantly with TSP-1 treatment. Western blot showed significantly reduced expression of TSP-1 in GzmB-treated retinal pigment epithelial cell culture and CSA supernatant compared with that in controls. Together, our findings suggest that the proteolysis of antiangiogenic factors such as TSP-1 by extracellular GzmB might represent a mechanism through which GzmB may contribute to nAMD-related CNV. Future studies are needed to investigate whether pharmacologic inhibition of extracellular GzmB can mitigate nAMD-related CNV by preserving intact TSP-1.

[Advances in the classification and treatment of isolated superior mesenteric artery dissection].

Isolated superior mesenteric artery dissection (ISMAD) has attracted more and more clinicians' attention in recent years. Patients onset of ISMAD often present with abdominal pain. The misdiagnosis or miss diagnosis is common because of the non-specific symptoms and signs, which even can endanger lives in serious cases. Imaging classification is of great significance for diagnosis and treatment of ISMAD. The Sakamoto classification and the Yun classification are two classical classified methods. However, with the further study of ISMAD, various new classifications emerge. Conservative treatment was once considered as the preferred. As the rapid development of endovascular therapy and the great progress of new devices, stenting therapy can significantly improve symptoms and achieve satisfactory long-term effects, and be even expected to become the preferred method for clinical therapy of ISMAD. However, the long-term effects of endovascular therapy still need a large number of follow-up data, and complications after stent implantation can't be ignored.

[Comparison of three dimensional CT venography and ascending phlebography based on propensity score matching in the evaluation of recurrent varicose veins].

Objective: To compare the efficacy of lower extremity three dimensional CT venography (CTV) and lower extremity ascending phlebography in evaluating recurrent varicose veins. Methods: A retrospective analysis was conducted on clinical data from 235 patients with unilateral recurrent varicose veins who were treated at the Department of Vascular Surgery,Beijing Shijitan Hospital,Capital Medical University, between January 2015 and December 2020.There were 112 males and 123 females, with an age of (62.5±11.4)years (range:24 to 75 years).Patients were stratified into two groups based on preoperative imaging examination:the CTV group (utilizing lower extremity venous ultrasound+lower extremity CTV) and the control group (employing lower extremity venous ultrasound+lower extremity ascending phlebography).The two groups were matched in a 1∶1 ratio using propensity score matching, resulting in 43 cases per group.Comparative analyses between the groups at the one-year postoperative follow-up were performed using independent sample t tests, Wilcoxon rank-sum tests, χ2 tests, and linear regression analysis. Results: One year post-surgery,the CTV group exhibited a lower venous clinical severity score (VCSS) compared to the control group(M(IQR),3.0(4.3) vs.4.0(5.8),Z=-2.038,P=0.040).Additionally, the chronic venous insufficiency patients' quality of life questionnaire (CIVIQ-20) scores were significantly higher in the CTV group than in the control group (89.0(8.0) vs.82.5(17.0), Z=-2.627, P=0.010).Patients in the CTV group also experienced a shorter ulcer healing time compared to the control group (4.0(4.0) weeks vs.12.0(7.0) weeks, Z=-3.217,P<0.01).Both groups showed no clinically symptomatic recurrent varicose veins or ulcers.However, they exhibited ultrasound-detectable varicose vein recurrence, with no statistically significant difference (χ2=0.453,P=0.500).The number of diseased vessels requiring management based on ultrasound supplemented by CTV was 16, while the number supplemented by ascending phlebography was 7,with a statistically significant difference (χ2=4.800,P=0.030).Linear regression analysis demonstrated that clinical-etiology-anatomy-pathology clinical grading and the preoperative imaging examination method exerted independent influences on VCSS and CIVIQ-20 during the one-year postoperative assessment. Conclusions: CTV-assisted ultrasound enables a direct and comprehensive evaluation and localization of diseased veins in patients with recurrent varicose veins.The utilization of lower extremity vein ultrasound combined with CTV-guided management of lower extremity vessels in minimally invasive treatment significantly improves patient prognosis, surpassing the assessment provided by ascending phlebography.

[The efficacy of thin struct bare stents for the treatment of spontaneous isolated dissection of the superior mesenteric artery].

Objective: To examine the safety and effectiveness of thin struct bare stents for the treatment of spontaneous isolated dissection of the superior mesenteric artery (SIDSMA). Methods: The data of 32 patients admitted to First Hospital of Jiaxing (20 cases) and Jinling Hospital (12 cases) with SIDSMA from January 2016 to January 2021 were retrospectively analyzed. There were 27 males and 5 females, aging (54.8±9.4) years (range: 36 to 75 years). All patients were treated with thin struct bare stents. Controllable spring coils were used to fulfill the false lumen in 2 cases. Symptoms, vascular remodeling pattern at the SIDSMA lesion, and patency of the stents were observed during follow-up. Results: The surgical success rate was 100%. According to the length of the lesions and stents, the number of stents implanted was 1 in 17 cases, 2 in 11 cases and 3 in 4 cases. The angiography showed that blood flow in the stent was smooth and that the false lumen disappeared or weakened. The numerical rating scale for abdominal pain decreased from 6.1±1.5 (range: 4 to 10) preoperatively to 1.0 (1.0) (range: 0 to 3) 1 hour postoperatively (W=528, P<0.01). The compression rate of the true lumen of the superior mesenteric artery decreased from (92.3±6.7)% (range: 25% to 94%) preoperatively to 0.8 (1.2)% (range: 0 to 3.2%) 1 month postoperatively (W=528, P<0.01). The primary patency rate of CT angiography at 1 month postoperatively was 100%. The vascular remodeling rate was (92.3±6.7)% (range: 80% to 100%). All patients were followed for (46.3±17.0) months (range: 24 to 76 months). The cumulative patency rates in 1, 2 and 5 years were all 100%. Conclusion: The use of thin struct bare stents for SIDSMA is safety and efficacy.

[Risk prediction of metabolic syndrome and coronary artery disease in overweight and obese populations based on serum metabolomics].

Objective: By identifying different metabolites in the serum and clarifying the potential metabolic disorder pathways in metabolic syndrome (MS) and stable coronary artery disease patients, to evaluate the predictive value of specific metabolites based on serum metabolomics for the occurrence of MS and coronary heart disease in overweight or obese populations. Methods: This is a retrospective cross-sectional study. Patients with Metabolic Syndrome (MS group), patients with stable coronary heart disease (coronary heart disease group), and overweight or obese individuals (control group) recruited from the Central District of the First Affiliated Hospital of Zhengzhou University from 2017 to 2019 were assigned to the training set, meanwhile, the corresponding three groups of people recruited from the East District of the hospital during the same period were assigned to the validation test. The serum metabolomics profiles were determined by ultra-performance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Clinical characteristics (age, gender, body mass index (BMI), blood pressure, fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), alanine aminotransferase (ALT), aspartate transaminase (AST), total cholesterol (TC), triacylglycerol (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), glomerular filtration rate (eGFR), creatinine (CR)) were also collected. Based on the orthogonal partial least-squares discrimination analysis (OPLS-DA) model, the significantly changed metabolites for MS and coronary artery disease patients were screened according to variable important in projection (VIP), and the receiver operating characteristic (ROC) analysis was evaluated for the risk prediction values of changed metabolites. Results: A total of 488 subjects were recruited in this study, the training set included 40 MS, 249 coronary artery disease patients and 148 controls, the validation set included 16 MS, 18 coronary artery disease patients and 17 controls. We made comparisons of the serum metabolites of coronary artery disease vs. controls, MS vs. controls, and coronary artery disease vs. MS, and a total of 22 different metabolites were identified. The disturbed metabolic pathways involved were phospholipid metabolism, amino acid metabolism, purine metabolism and other pathways. Through cross-comparisons, we identified 2 specific metabolites for MS (phosphatidylcholine (18∶1(9Z)e/20) and pipecolic acid), 4 specific metabolites for coronary artery disease (lysophosphatidylcholine (17∶0), PC(16∶0/16∶0), hypoxanthine and histidine), and 4 common metabolites both for MS and coronary artery disease (isoleucine, phenylalanine, glutathione and LysoPC(14∶0)). Based on the cut-off values from ROC curve, the predictive value of the above metabolites for the occurrence of MS in overweight or obese populations is 100%, the predictive value for the occurrence of coronary heart disease is 87.5%, and the risk predictive value for coronary heart disease in MS patients is 82.1%. Conclusions: The altered serum metabolites suggest that MS and coronary heart disease may involve multiple metabolic pathway disorders. Specific metabolites based on serum metabolomics have good predictive value for the occurrence of MS and coronary heart disease in overweight or obese populations.

[Modified classification, screening, prevention and treatment of cardioembolic stroke].

Ischemic stroke is usually regarded as a neurological disease, but cardiogenic cerebral embolism is actually one of the most common identifiable causes of ischemic stroke. According to the origin of embolus (left heart/right heart system) and related structural abnormalities, this article expanded and refined the classification definition of cardiogenic cerebral embolism, and proposed an improved classification method of ischemic stroke (PUMCH-ISC). In addition, this article gave a practical clinical screening program for cardiovascular physicians according to different causes of stroke, and sorted out the prevention and treatment strategies and progress of common causes of cardiogenic cerebral embolism, aiming to help cardiovascular physicians and neurologists improve the screening, diagnosis and accurate prevention and treatment of cardiogenic cerebral embolism.

[The application value of cardiac magnetic resonance quantitative T1 mapping technique for risk stratification in patients with pulmonary arterial hypertension].

Objective: To explore the application value of cardiac magnetic resonance (CMR) native T1 mapping for risk stratification in patients with pulmonary arterial hypertension (PAH). Methods: A total of 59 patients with diagnosed PAH and clear-documented risk status in Peking Union Medical College Hospital and underwent CMR examination between January 2019 and December 2021 were retrospectively included, which including 12 males and 47 females, aged from 4 to 77 (31±13) years. Those patients were subdivided into two groups based on the clinically-assessed risk status: low-risk group (n=30) and intermediate-/high-risk group (n=29). Twenty-five healthy individuals were included as controls. Base, midventricular, and apical inferior right ventricular insertion point (IRVIP) native T1 values on short axis images were measured. Native T1 values in PAH patients and control group, in low-risk group and intermediate-/high-risk group were compared, respectively, and receiver operating characteristics (ROC) curves with area under the curves (AUC) were calculated to evaluate the application value of native T1 values for risk stratification in PAH patients. Results: Base, midventricular and apical IRVIP native T1 of PAH patients were all significantly increased as compared to controls [Base:(1 439.31±129.96) vs (1 282.36±37.18) ms;midventricular:(1 450.32±111.55) vs (1 287.56±53.16) ms;apical:(1 444.12±109.15) vs (1 266.36±75.31) ms](all P<0.001). The midventricular IRVIP native T1 values were significantly higher in patients in intermediate-/high-risk status as compared to those in low-risk status [ (1 493.24±126.32) vs (1 428.50±85.73) ms,P=0.026]. The AUC of mid ventricle IRVIP native T1 for distinguishing patients in intermediate-/high-risk status was 0.741. The base [(1 458.21±134.96) vs (1 421.03±104.75) ms, P=0.241] and apical [(1 465.90±125.36) vs (1 423.07±87.87) ms,P=0.136] IRVIP native T1 values in patients in intermediate-/high-risk group were also numerically higher as compared with patients in low-risk status, however, without statistical significant (both P>0.05). Conclusion: Midventricular IRVIP native T1 value might have a role for assisting in risk stratification in PAH patients, which was clinically significant for facilitating the work-up and prognosis improvement of PAH patients.

[A Luciferase-EGFP Reporter System for the Evaluation of DNA Methylation in Mammalian Cells].

DNA methylation is an essential epigenetic modification involved in numerous biological processes. Here, we present a cell-based system pLTR-Luc2P-EGFP for evaluation of DNA methylation in mammalian cells. In this system, the expression of reporter gene luciferase2P (Luc2P)-EGFP is under the control of HIV-1 promoter 5' long terminal repeat (LTR), which contains multiple CpG sites. Once these sites are methylated, the expression of Luc2P-EGFP is turned off, which may be visualized under fluorescence microscopy, with quantification performed in luciferase activity assay. As a proof of principle, pLTR-Luc2P-EGFP was methylated in vitro, and transfected into 293T cells, where the reduction of Luc2P-EGFP expression was confirmed. Premixed reporter DNA samples with the methylation levels varying from 0 to 100% were used for quantitative measurements of DNA methylation. The resulting standard curves indicated the accuracy of luciferase activity exceeding that of the Western blotting against EGFP. The Bland-Altman analysis showed that data from luciferase activity assay were in good agreement with the actual DNA methylation levels. In summary, we have established a reporter system coupled with reliable detection technique capable of efficient quantifying the changes in methylation in mammalian cells. This system may be utilized as a high throughput screening tool for identifying molecules that modulate DNA methylation.

[Feasibility and safety of right ventricular endomyocardial biopsy via the right internal jugular vein approach].

Objective: To evaluate the feasibility and safety of right ventricular endomyocardial biopsy (EMB) via the right internal jugular vein approach. Methods: It was a retrospective and descriptive study. A total of 272 patients, who underwent right ventricular EMB from December 2014 to June 2020 in Fuwai Hospital and Peking Union Medical College Hospital were enrolled. The preliminary diagnosis included suspected myocarditis, myocardiopathy, unexplained heart failure etc after exclusion of coronary heart disease. Clinical characteristics including age, sex, height, weight, NYHA functional class, NT-proBNP, chest radiography, echocardiography, and hemodynamics parameters were collected at baseline. EMB was performed via right internal jugular vein approach under the biplane fluoroscopic guidance. Success rate was calculated in this study. Complications related to operation were recorded according the following definitions. Major complications included death, urgent cardiac surgery, advanced cardiac life support, pericardiocentesis in cardiac tamponade, permanent complete atrioventricular block requiring permanent pacing etc. Minor complications included pericardial effusion without pericardiocentesis, temporary (lasting less than 24 hours) or permanent right bundle-branch block, temporary Mobitz type Ⅱ atrioventricular block (AV block) with AV conduction 2∶1 requiring medical treatment with atropine, or additive temporary pacing, non-sustained ventricular tachycardia with long runs of more than 10 ventricular complexes, and an episode of atrial fibrillation lasting less than 12 hours or cardioversion of atrial fibrillation. Other complications included tricuspid anterior chorda rupture and new onset tricuspid regurgitation after EMB. Results: In this study, right ventricular EMB were performed successfully in 270 patients, the total success rate was 99.3% (270/272), and EMB were failed in 2 (0.7%) patients. Age of the enrolled patients was (42.7±16.9) years, and there were 164 (60.3%) males. Major complication including cardiac tamponade requiring pericardiocentesis occurred in 2 (0.7%) patients. Minor complications such as small amount pericardial effusion occurred in 18 (6.6%) patients, tricuspid anterior chorda rupture occurred in 1 (0.4%) patient. No patient died, or requiring permanent pacing, or requiring emergency cardiac surgery. The complication rate was 9.3% (13/140), 7.8% (7/90), and 2.4% (1/42) in operators with 1, 2, and 3 years' experience. Conclusions: EMB via the right jugular vein approach under fluoroscopic guidance is a simple, safe and feasible procedure. The complication rates decrease significantly with increasing operator experience.

[Feasibility of the determination of plasma vardenafil level in rat by performance liquid chromatography-tandem mass spectrometry].

Objective: To develope and validate a reliable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of vardenafil concentration in plasma of rat. Methods: Plasma samples of normal Sprague-Dawley rats were collected. A Phenomenex Synergi Polar-RP 80A column (2.0 mm×50 mm, 4 µm) was used. Column temperature was set at 30 ℃. Mobile phase A was 0.1% formic acid in water; mobile phase B was 0.1% formic acid in acetonitrile. The flow rate was 0.4 ml/minutes. Quantitative determination was performed by electrospray ionization, operating in positive ion multiple reaction monitoring (MRM) mode. Cisapride was used as the internal standard. The feasibility of the method was evaluated by examining its specificity, linearity and quantitative range, precision and accuracy, matrix effects, and stability. Results: Under the selected chromatographic and mass spectrometry conditions, the monitoring ions of vardenafil and internal standard were mass-to-charge ratio（m/z) 489.3/151.2 and 466.4/234.2, the retention times of vardenafil and internal standard were 2.62 and 2.80 minutes, respectively, and the peak shape was satisfactory. The method has good linearity in the concentration range of 0.2-200 ng/ml. The intra-batch precision (%CV) and accuracy (%DEV) of vardenafil were 1.5%-9.7% and -6.8%-6.6%, respectively. The inter-batch precision and accuracy of vardenafil were 3.1% -8.4% and -3.7%-4.6%, respectively. In this sample processing method, the extraction recovery rate of vardenafil was obtained at range of 88.2%-104.6%, which met the requirements for the investigation of extraction recovery rate. In this sample processing method, the normalized matrix factor of each quality control concentration of vardenafil was 1.04, 0.85, and 1.04, and the coefficient of variation (%CV) was in the range of 1.7%-10.7%, which met the requirements for the investigation of matrix effects. Variations of short-term stability, long-term stability, and stability of 4 freeze-thaw cycles of vardenafil was within ±15%, and the coefficient of variation were within 5%. Conclusion: The high performance liquid chromatography-tandem mass spectrometry method established in this study is feasible for the measurement of concentration of vardenafil in rat plasma and this method has good specificity and high accuracy, and can be used to detect the concentration of vardenafil in rat plasma.

The reduction of XIAP is associated with inflammasome activation in RPE: implications for AMD pathogenesis.

BACKGROUND: Age-related macular degeneration (AMD) is a multifactorial chronic disease of the eye. Several candidate pathways have been hypothesized to play a role in AMD pathogenesis. Our work and those of others suggests inflammasome activity as a mechanism associated with retinal pigment epithelial (RPE) cell demise. X-linked inhibitor of apoptosis protein (XIAP), an anti-apoptosis factor, has recently been shown to regulate inflammasome activity in non-ocular cells. The purpose of this study is to characterize XIAP's regulatory role in RPE. METHODS: Protein lysates of eye tissues from rats (vinpocetine- or aurin tricarboxylic acid complex-treated, ATAC, vs naïve) and mice (wild type vs Caspase-4-/-) were utilized to analyze XIAP protein levels. Immunohistochemistry was used to detect NLRP3 levels in the RPE layer. In vitro inflammasome activation on RPE cells was achieved with L-leucyl-L-leucine methyl ester (Leu-Leu-OMe) stimulation. Levels of XIAP mRNA and 18S RNA were quantified by RT-PCR. Cell culture supernatants were tested directly for secreted IL-1ß by ELISA or concentrated for the detection of secreted IL-18 by western blot. Protein lysates from RPE in cell culture were collected for the measurement of cleaved caspase-1 p20, XIAP, and GAPDH. Data are presented as Mean ± SD. p < 0.05 is considered statistically significant. RESULTS: The XIAP protein level was significantly increased when the inflammasome was inhibited at the "activation" step by ATAC, but not the "priming" step, in vivo. Concomitantly, NLRP3 immunoreactivity was lower in the RPE layer of animals fed with ATAC. In mice where caspase-1 cleavage was impaired by the genetic deficiency in caspase-4, the XIAP protein level increased in eye tissues. In RPE cell culture, Leu-Leu-OMe stimulation led to caspase-1 cleavage, cytokine secretion, and XIAP reduction, which can be abolished by Z-YVAD-FMK. When XIAP siRNA was given as a pre-treatment to RPE in vitro, Leu-Leu-OMe induced IL-1ß/IL-18 secretion was enhanced, whereas overexpressing XIAP reduced IL-1ß secretion under inflammasome activation, both compared to controls cells. CONCLUSIONS: Together, these data suggest XIAP-mediated inhibition of inflammasome activity in RPE may provide insights into the biological consequences of inflammasome activation in RPE and reveals the caspase-1/XIAP/IL-1ß/IL-18 axis as a target for broader applications in AMD biology and treatment design.

[Advances in the effects on false lumen thrombosis after the endovascular treatment of aortic dissection and its intervention strategies].

Aortic dissection is a urgent and dangerous disease, and the endovascular treatments can cure it in an effective and efficient way. As an important indicator of aorta remodeling, false lumen thrombosis is related to the prognosis of aortic dissection after endovascular treatment. The relationships among false lumen thrombosis after endovascular treatment and prognosis of aortic dissection, and the clinical techniques that can promote the false lumen thrombosis are reviewed in this paper. The aim is to promote a comprehensive evaluation of aorta remodeling.

Evidence for the activation of pyroptotic and apoptotic pathways in RPE cells associated with NLRP3 inflammasome in the rodent eye.

BACKGROUND: Age-related macular degeneration (AMD) is a devastating eye disease causing irreversible vision loss in the elderly. Retinal pigment epithelium (RPE), the primary cell type that is afflicted in AMD, undergoes programmed cell death in the late stages of the disease. However, the exact mechanisms for RPE degeneration in AMD are still unresolved. The prevailing theories consider that each cell death pathway works independently and without regulation of each other. Building upon our previous work in which we induced a short burst of inflammasome activity in vivo, we now investigate the effects of prolonged inflammasome activity on RPE cell death mechanisms in rats. METHODS: Long-Evans rats received three intravitreal injections of amyloid beta (Aß), once every 4 days, and were sacrificed at day 14. The vitreous samples were collected to assess the levels of secreted cytokines. The inflammasome activity was evaluated by both immunohistochemistry and western blot. The types of RPE cell death mechanisms were determined using specific cell death markers and morphological characterizations. RESULTS: We found robust inflammasome activation evident by enhanced caspase-1 immunoreactivity, augmented NF-κB nuclear translocalization, increased IL-1ß vitreal secretion, and IL-18 protein levels. Moreover, we observed elevated proteolytic cleavage of caspase-3 and gasdermin D, markers for apoptosis and pyroptosis, respectively, in RPE-choroid tissues. There was also a significant reduction in the anti-apoptotic factor, X-linked inhibitor of apoptosis protein, consistent with the overall changes of RPE cells. Morphological analysis showed phenotypic characteristics of pyroptosis including RPE cell swelling. CONCLUSIONS: Our data suggest that two cell death pathways, pyroptosis and apoptosis, were activated in RPE cells after exposure to prolonged inflammasome activation, induced by a drusen component, Aß. The involvement of two distinct cell death pathways in RPE sheds light on the potential interplay between these pathways and provides insights on the future development of therapeutic strategies for AMD.

Transcatheter aortic valve replacement with balloon-expandable valve : Analysis of initial experience in China.

BACKGROUND: Transcatheter aortic valve replacement (TAVR) is widely applied for the treatment of severe aortic stenosis (AS) in developed countries; however, in China, it is still in the early stage of utilization. On the basis of previous studies, this work explored the feasibility of TAVR in patients with severe AS in China and analyzed the cause of death in four cases. METHODS: This retrospective study included 20 patients who had severe AS and underwent TAVR with a balloon-expandable system (Edwards SAPIEN XT) in our hospital from January 2011 to June 2016. The valve and heart functions of 16 survivors before and after the TAVR procedure were compared. TAVR endpoints, device success, and adverse events were assessed according to the definitions of the Valve Academic Research Consortium-2 (VARC-2). RESULTS: There were 13 male and seven female patients aged 65-81 years (average, 73.15) who underwent TAVR. The TAVR approach was transfemoral in 19 patients and transapical in one patient. Four patients died (two of coronary artery occlusion and two of aortic annulus rupture) during the TAVR procedure or shortly after; six patients had mild paravalvular leakage, and the rest of the patients showed a significant improvement in cardiac function. During the follow-up period (2-62 months), one patient died of lung cancer 13 months after the TAVR procedure. CONCLUSION: TAVR with a balloon-expandable system is safe and effective and can be used for patients with severe AS in China. It requires careful patient selection and preoperative assessment so as to reduce the 30-day postoperative mortality rate.

Indoor particle dynamics in a school office: determination of particle concentrations, deposition rates and penetration factors under naturally ventilated conditions.

Recently, the problem of indoor particulate matter pollution has received much attention. An increasing number of epidemiological studies show that the concentration of atmospheric particulate matter has a significant effect on human health, even at very low concentrations. Most of these investigations have relied upon outdoor particle concentrations as surrogates of human exposures. However, considering that the concentration distribution of the indoor particulate matter is largely dependent on the extent to which these particles penetrate the building and on the degree of suspension in the indoor air, human exposures to particles of outdoor origin may not be equal to outdoor particle concentration levels. Therefore, it is critical to understand the relationship between the particle concentrations found outdoors and those found in indoor micro-environments. In this study, experiments were conducted using a naturally ventilated office located in Qingdao, China. The indoor and outdoor particle concentrations were measured at the same time using an optical counter with four size ranges. The particle size distribution ranged from 0.3 to 2.5 µm, and the experimental period was from April to September, 2016. Based on the experimental data, the dynamic and mass balance model based on time was used to estimate the penetration rate and deposition rate at air exchange rates of 0.03-0.25 h-1. The values of the penetration rate and deposition velocity of indoor particles were determined to range from 0.45 to 0.82 h-1 and 1.71 to 2.82 m/h, respectively. In addition, the particulate pollution exposure in the indoor environment was analyzed to estimate the exposure hazard from indoor particulate matter pollution, which is important for human exposure to particles and associated health effects. The conclusions from this study can serve to provide a better understanding the dynamics and behaviors of airborne particle entering into buildings. And they will also highlight effective methods to reduce exposure to particles in office buildings.

[Primary investigation of immunoglobulin A synthesis and secretion in human mesangial cells].

OBJECTIVE: To investigate the expression of immunoglobulin A (IgA) in human mesangial cells (HMCs). METHODS: The HMCs were cultured. The subcellular location of IgA was detected by immunofluorescence staining; the transcripts of Igα,Igκ and IgΛ constant region were detected by reverse transcription-polymerase chain reaction (RT-PCR) and further analyzed by DNA sequencing. The expressions of Igα and IgΛ were detected at transcription level by Western blot after the cytoplasmic protein extraction. The culture supernatant was collected to explore whether IgA could be secreted out of the cell and the protein was further analyzed by mass spectrometry after being purified by affinity chromato-graphy with jacalin-sepharose. The results of DNA sequencing and mass spectrometry were aligned with the mRNA and amino acid sequences in the National Center of Biotechnology Information (NCBI) database. RESULTS: By immunofluorescence staining, we detected the presence of IgA heavy chain Igα, light chain, both Igκ and IgΛ in expressions of transcripts of Igα1, Igα2, Igκ and IgΛ in the HMCs and the alignment of the sequences of the RT-PCR products with those of the IgCα1, IgCα2, Igκ and IgΛ mRNA in the NCBI database exhibited that the similarities were 99%, 97%, 98% and 97%, respectively. Western blot showed Igα and IgΛ expressions in the cell lysate and secretion of Igα1 and Igα2 heavy chains in cell culture supernatant. To further explore the protein that secreted into the supernatant, after supernatant affinity chromatography with jacalin-sepharose, the proteins were separated by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) and the band approximating to 65 000 was cut and sent to mass spectrometry. The results were aligned with the amino acid sequences of Igα1 and Igα2 constant region in NCBI database, showing that amino acids between No.52 and No.104, amino acids between No.154 and No.221, amino acids between No.276 and No.327 from IgCα1 and amino acids between No.52 and No.113, amino acids between No.151 and No.204, amino acids between No.251 and No.314 from IgCα2 were the same with those derived from B cells. CONCLUSION: Our fin-dings suggested that HMCs could synthesize and secret IgA.