Retinopathy of prematurity (ROP) is a vision-threatening disorder that leads to pathological growth of the retinal vasculature due to hypoxia. Here, we investigated the potential effects of alamandine, a novel heptapeptide in the renin-angiotensin system (RAS), on hypoxia-induced retinal neovascularization and its underlying mechanisms. In vivo, the C57BL/6J mice with oxygen-induced retinopathy (OIR) were injected intravitreally with alamandine (1.0 μmol/kg per eye). In vitro, human retinal microvascular endothelial cells (HRMECs) were utilized to investigate the effects of alamandine (10 μg/mL) on proliferation, apoptosis, migration, and tubular formation under vascular endothelial growth factor (VEGF) stimulation. Single-cell RNA sequencing (scRNA-seq) matrix data from the Gene Expression Omnibus (GEO) database and RAS-related genes from the Molecular Signatures Database (MSigDB) were sourced for subsequent analyses. By integrating scRNA-seq data across multiple species, we identified that RAS-associated endothelial cell populations were highly related to retinal neovascularization. The liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis revealed a significant decrease in alamandine levels in both the serum and retina of OIR mice compared to those in the control group. Next, alamandine ameliorated hypoxia-induced retinal pathological neovascularization and physiologic revascularization in OIR mice. In vitro, alamandine effectively mitigated VEGF-induced proliferation, scratch wound healing, and tube formation of HRMECs primarily by inhibiting the hypoxia-inducible factor-1α (HIF-1α)/VEGF pathway. Further, coincubation with D-Pro⁷ (Mas-related G protein-coupled receptor D (MrgD) antagonist) hindered the beneficial impacts of alamandine on hypoxia-induced pathological angiogenesis both in vivo and in vitro. Our findings suggested that alamandine could mitigate retinal neovascularization by targeting the MrgD-mediated HIF-1α/VEGF pathway, providing a potential therapeutic agent for OIR prevention and treatment.
Animals ; Retinal Neovascularization/prevention & control* ; Mice, Inbred C57BL ; Vascular Endothelial Growth Factor A/metabolism* ; Humans ; Mice ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Oligopeptides/therapeutic use* ; Signal Transduction/drug effects* ; Cell Proliferation/drug effects* ; Endothelial Cells/drug effects* ; Retinopathy of Prematurity/drug therapy* ; Apoptosis/drug effects* ; Cell Movement/drug effects* ; Renin-Angiotensin System/drug effects* ; Cells, Cultured

Objective:To investigate the application value of virtual simulation experiment in the teaching of Laboratory Sciences for Virus Infection Testing.Methods:Undergraduate students(n=58)majoring in Health Inspection and Quarantine of grade 2020 were selected as the research objects.The"Virtual simulation experiment of influenza virus isolation and identification in BSL-2 laboratory"was included in the teaching of Laboratory Sciences for Virus Infection Testing.The application effect of the virtual simulation experiment was evaluated by a self-designed questionnaire.Results:A total of 50 students completed the questionnaire effectively.After completing the virtual simulation experiment,96.0%of the students thought that the simulation degree of the experiment was high.Students could understand the instrument composition,internal structure,operation process,and matters needing attention.And 52.0%of the students thought that the experiment had an impact on autonomous learning.Students believed that the experiment could improve their interest in the course(88.0%),increase their mastery of theoretical knowledge(60.0%),and improve their ability to analyze and solve problems(46.0%).And 72.0%of the students thought that the running speed of the virtual simulation experiment was slow.The top three advantages of the experiment evaluated by students were easy operation,no damage to the instrument and no worry about the dangers in the experiment,and helpful to understand the knowledge.The top three disadvantages of this experiment were no improvement in practical ability,no familiarity with the use of instruments,different feelings from the real experiment and too idealized experimental process.And 66.0%of the students thought that virtual simulation experiments should complement traditional experimental teaching.Conclusions:Virtual simulation experiment can carry out dangerous experiments that are difficult to perform in traditional laboratories.It can improve students'mastery of theoretical knowledge,enhance their interest in learning,and improve their ability to analyze and solve problems.However,there are still some problems in its application,which should be combined with traditional experimental teaching to improve the teaching effect.

Background: To evaluate the feasibility of treating odontoid fractures in the Chinese population with two cortical screws based on computed tomography (CT) scans and describe a new measurement strategy to guide screw insertion in treating these fractures. Methods: A retrospective review of cervical computed tomographic scans of 128 patients (aged 18–76 years; men, 55 [43.0%]) was performed. The minimum external transverse diameter (METD), minimum external anteroposterior diameter (MEAD), maximum screw length (MSL), and screw projection back angle (SPBA) of the odontoid process were measured on coronal and sagittal CT images. Results: The mean values of METD and MEAD were 10.0 ± 1.1 mm and 12.0 ± 1.0 mm, respectively, in men and 9.2 ± 1.0 mm and 11.0 ± 1.0 mm, respectively, in women. Both measurements were significantly higher in men (p < 0.001). In total, 87 individuals (68%) had METD > 9.0 mm that could accommodate two 3.5-mm cortical screws. The mean MSL value and SPBA range were 34.4 ± 2.9 mm and 13.5°–24.2°, respectively, with no statistically significant difference between men and women. Conclusions The insertion of two 3.5-mm cortical screws was possible for anterior fixation of odontoid fractures in 87 patients (68%) in our study, and there was a statistically significant difference between men and women.

ObjectiveTo study the effect of Bazi Bushen capsules on delaying the aging process of naturally aging mice and its mechanism. MethodThe mice were randomly divided into four groups according to their body weight， namely， aging group， low-dose Bazi Bushen capsules group （1 g·kg^-1）， high-dose Bazi Bushen capsules group （2 g·kg^-1）， and rapamycin group （0.002 g·kg^-1）. The debilitating signs were detected by behavioral tests and the weakness index was measured. The percentages of spleen T and B lymphocytes， effector T cells （TE）， memory T cells （TM）， naive T cells （TN）， helper T cells （Th）， cytotoxic T cells （Tc） ，Th1 cells， Th2 cells， and regulatory T cells （Treg） were determined by flow cytometry. Cell proliferation and the cell counting kit-8 （CCK-8） assay was used to detect the proliferation of lymphocytes in mice. The hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the mouse spleen. The expression of cyclin-dependent kinase inhibitor 2A （p16） and cyclin-dependent kinase inhibitor 1A （p21） was detected by immunohistochemistry （IHC）. The mRNA expression of senescence-related proteins p16 and p21 was determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the inflammatory cytokines， including tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin （IL）-1β， IL-2， IL-4， IL-10， and IL-12 p70， in peripheral blood of mice were detected by Luminex. ResultAs compared with the aging group， mice in the Bazi Bushen capsules and rapamycin groups showed significantly improved debilitating signs and reduced weakness index scores （P<0.05， P<0.01）， increased proportions of T cells， TN cells， Tc cells， Th2 cells， and Treg cells in the spleen， decreased proportions of TE cells， TM cells， Th cells， Th1 cells （P<0.05， P<0.01）， and increased proliferation of splenic lymphocytes （P<0.05， P<0.01）. In the Bazi Bushen capsules and rapamycin groups， clear structure of the red and white marrow marginal zone was observed in the spleen of mice， the area of the white marrow was increased， and the area of the red marrow was correspondingly decreased. The protein and mRNA expression of aging-related proteins p16 and p21 in the spleen was decreased （P<0.01）， the levels of serum pro-inflammatory cytokines IL-1β， IL-2， IL-12 p70， IFN-γ， and TNF-α levels were decreased， and the levels of IL-4 and IL-10 were increased （P<0.05， P<0.01） in the Bazi Bushen capsules and rapamycin groups as compared with the aging group. ConclusionBazi Bushen capsules have the effect of regulating the debilitating signs of natural aging mice， regulating the immune homeostasis and inflammation level of the body， and reducing cell aging.

Development of thoracolumbar vertebra (TLV) and rib primordium (RP) is a common evolutionary feature across vertebrates, although whole-organism analysis of the expression dynamics of TLV- and RP-related genes has been lacking. Here, we investigated the single-cell transcriptome landscape of thoracic vertebra (TV), lumbar vertebra (LV), and RP cells from a pig embryo at 27 days post-fertilization (dpf) and identified six cell types with distinct gene expression signatures. In-depth dissection of the gene expression dynamics and RNA velocity revealed a coupled process of osteogenesis and angiogenesis during TLV and RP development. Further analysis of cell type-specific and strand-specific expression uncovered the extremely high level of HOXA10 3'-UTR sequence specific to osteoblasts of LV cells, which may function as anti-HOXA10-antisense by counteracting the HOXA10-antisense effect to determine TLV transition. Thus, this work provides a valuable resource for understanding embryonic osteogenesis and angiogenesis underlying vertebrate TLV and RP development at the cell type-specific resolution, which serves as a comprehensive view on the transcriptional profile of animal embryo development.

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

Objective To investigate the correlation between quantitative perfusion histogram parameters of DCE-MRI and tumor tissue microvessel density(MVD) in patients with lung cancer.Methods 30 patients with lung cancer confirmed by pathology who underwent preoperative DCE-MRI were enrolled in this retrospective study .Quantitative perfusion histogram pa-rameters( including median, mean, skewness, kurtosis, energy, entropy) were measured for each patient using Exchange mo-dle.Using the Immunohistochemical method to detect the expression of CD34 in tumor tissue, and counting the number of mi-crovessels under microscope.SPSS 19.0 was used to carry out statistical analysis.The correlation between MVD and quantita-tive perfusion histogram parameters of DCE-MRI measured by exchange model was evaluated by Pearson linear correlation anal-ysis.Results There was no significant difference in MVD and each quantitative perfusion histogram parameters between the three different pathological groups of lung cancer(P >0.05).Ktrans perfusion histogram parameters(mean, 25％, 50％, 50％, 75％, 90％, 95％), Kep perfusion histogram parameters(entropy, 10％, 25％, 50％, 75％, 90％), Fp perfusion histogram parameters( mean, 25％, 50％, 75％, 90％, 95％) and Vp perfusion histograms parameters ( entropy, 75％, 90％, 95％ ) were positively correlated with MVD(P<0.05).Ktrans perfusion histogram parameters(energy) and Vp perfu-sion histogram parameters(skewness, kurtosis, energy) were negatively correlated with MVD(P<0.05).There was no signifi-cant correlation between Ve perfusion histogram parameters and MVD(P>0.05).Conclusion There was a certain correla-tion between the perfusion histogram parameters of DCE-MRI and MVD,suggesting that the quantitative perfusion histogram of DCE-MRI in lung cancer can reflect the MVD value of cancer tissue .

Objective To compare the efficacy of trimodality therapy and chemoradiation therapy (CRT) alone in patients with locally advanced resectable esophageal squamous cell carcinoma (SCC).Methods A total of 124 cases with locally advanced resectable esophageal SCC were retrospectively analyzed and classified into 2 groups.Fifty-four cases in trimodality group were treated with surgery and preoperative chemoradiation,while 70 cases in CRT alone group only received radiation and chemotherapy.Local tumor control,3-year survival and treatment-related mortality were assessed.Results The local recurrent rate of the resected patients was 18.5％ in trimodality group and 35.7％ in CRT alone group,respectively(x2 =4.445,P ＜ 0.05).The 3-year progression-free survival (PFS) was 65.3％ (95％ CI 50.7-80.5) in trimodality group and31.9％ (95％CI 19.6-44.2) in CRT alone group (P＜0.05),while the overall survival (OS) 66.3％ (95％ CI43.0-89.6) and 34.4％ (95％ CI 21.1-47.7),respectively(P ＜ 0.05).Treatment-related mortality was 1.9％ in trimodality group and 2.9％ in CRT alone group (P ＞ 0.05).For CRT alone group,the sub-group analysis showed that there was no statistically significant difference in the 3-year OS between patients who received 50-50.4 Gy and those who received the dose over 50.4 Gy (39.9％ 95％ CI 18.5-61.3 vs.31.5％ 95％ CI 14.8-48.2,P ＞0.05).Conclusions Compared with CRT alone,trimodality therapy showed the superior local control,PFS and OS,with similar treatment-related mortality in the treatment of patients with SCC of esophagus.The role of surgery could not be replaced by CRT alone even with the augment of radiation dose.

Objective To explore the clinical efficacy of nibble debridement combined with silver ions dressing in treatment of diabetic foot ulcer.Methods The data of 238 patients with diabetic foot ulcer in people's hospital of Meishan from January 2012 to December 2016 were retrospectively analyzed.Of which 125 patients were treated by nibble debridement combined with silver ions dressing(nibble debridement group),113 patients were treated with surgical debridement combined with silver ions dressing(surgical debridement group).The clinical efficacy,wound healing time and complication between two groups were compared.Results The wound healing time of nibble debridement group was (37.5 ± 10.8) days,which was shorter than (45.3 ± 11.7) days of surgical debridement group,the difference was significant (P ＜0.05).The total effective rate of nibble debridement group was 92.80％,which was higher than 73.45％ of surgical debridement group,the difference was significant(P ＜ 0.05).No complication occurred in two groups.Conclusion The method of nibble debridement combined with silver ions dressing treatment has a better satisfactory clinical result,shorter wound healing time and lower medical costs for patients with diabetic foot ulcer.

Objective To explore the distribution characteristics of ankle-brachial index ( ABI) examination for peripheral arterial dis-ease in diabetic patients, and to provide basis for clinical diagnosis. Methods Randomly selected 110 cases of patients who were admitted into our hospital from February 2014 to January 2015, and divided them into the observation group (34 cases with diabetic lower extremity peripheral vascular lesions) and the control group (76 cases without diabetic lower extremity peripheral vascular lesions). Measured the ABI of the two groups, and analyzed the value of ABI data range for the diagnosis of diabetic lower extremity peripheral vascular lesions. Mean-while, analyzed the risk factors of peripheral arterial disease in diabetic patients according to ABI. Results The sensitivity of ABI for diag-nosis of diabetic lower extremity peripheral vascular lesions was 91. 18%, and the coincidence rate was 95. 36%, the specificity was 98. 68%, the misdiagnosis rate was 8. 82%, and the omission diagnostic rate was 8. 82%. ABI of the observation group was obviously lower than that of the control group, and the difference was statistically significant (P<0. 05). Conclusion ABI examination could be basis of clinical diagnosis and important standard of disease assessment for peripheral arterial disease in diabetic patients. It is suggested to strengthen the control of blood glucose, blood pressure and weight of patients with diabetes in order to reduce the risk of morbidity.