Average Optical Density Research Articles

MicroRNA microarray analysis to detect biomarkers of aortic dissection from paraffin-embedded tissue samples.

The aim of this study was to explore the differential expression profiles of microRNAs (miRNAs) in paraffin-embedded acute aortic dissection (AAD) tissues to find potential biomarkers for this disease. A total of 92 paraffin-embedded tissue specimens were collected from 92 patients with AAD who underwent surgical replacement. Among these specimens, 54 had partial normal aortic segments (smooth intima surface, non-atherosclerotic lesions) in proximal crevasse of aorta. Samples of these segments were taken 1 cm away from aortic lesions as the control group, after eliminating the tunica adventitia tissues. miRNA expression profiles were obtained by miRNA microarray analysis. Differentially expressed miRNAs were found by comparing the AAD group with the control group and were verified by fluorescence real-time quantitative polymerase chain reaction and by fluorescence in situ hybridization. A total of 71 differentially expressed miRNAs were detected. Twenty-two were up-regulated and 49 were down-regulated. Four up-regulated miRNAs (hsa-miR-636, hsa-miR-142-3p, hsa-miR-425-3p, hsa-miR-191-3p) were selected for validation by real-time fluorescence quantitative polymerase chain reaction and fluorescence in situ hybridization. In the fluorescence real-time quantitative polymerase chain reaction analysis, only hsa-miR-636 showed a statistically significant difference in the AAD versus control comparison (3.3-fold, P = 0.012). The fluorescence in situ hybridization validation showed that the expression level of hsa-miR-636 was significantly increased in the AAD versus control comparison (P < 0.001), with average optical densities of 61.29 ± 16.83 in the AAD group and 9.30 ± 3.98 in the control group. Hsa-miR-636 is involved in the pathogenesis of AAD and may be a potential biomarker for this disease.

Effect of Buddleja officinalis eye drops to inflammatory factors of lacrimal gland cells of castrated male rabbit with dry eye

AIM: To observe the effect of different concentrations Buddleja offcinalis eye drops (BOED) to inflammatory factors of lacrimal gland cells of castrated male rabbit with dry eye. METHODS: Forty-two healthy adult New Zealand rabbits were randomly divided into 7 groups, 6 rabbits each group. Group A: blank group, Group B: model group, Group C: low concentrations BOED group, Group D: medium concentrations BOED, Group E: high concentrations BOED, Group F: the placebo group, Group G: testosterone groups. All rabbits of group B to G were cut off bilateral testis and epididymis except group A. The rabbits of group C, D, E and F were dropped relevant eye drops except groups A and B. All rabbits were tested Schimer I test (SIT) and break-up time (BUT) before operation and 4wk after operation. Expressions of inflammatory factors of TNF-α, IL-1β from lacrimal gland cells were checked in immunohistochemical staining method. RESULTS: 1) Comparison of SIT and BUT: Compared with the groups D, E and G, group C had statistical significance (P&lt;0.01). Compared with group D and E, G groups, there was no significant difference among those group before and after operation (P&gt;0.05); 2) Comparison of inflammatory factors of TNF-α, IL-1β after operation: Compared with value of the average optical density of TNF-α and IL-1β of group C, those of group D, E and G were obviously different after operation (P&lt;0.05). Compared D and with E, G groups, there was no significant difference between those group after operation (P&gt;0.05).

Efficacy Analysis of Bronchoscopic Interventional Therapy for Benign Tracheal Stenosis by Combination of Intelligent Imaging Analysis System and Immunohistochemistry

Objective: To assess the efficacy of bronchoscopic interventional therapy for benign tracheal stenosis by combination of intelligent imaging analysis system and immunohistochemistry. Methods: The therapeutic effect was evaluated by Computed Tomography (CT) image changes, and the effects of thermofrequency and glucocorticoids were analyzed by animal experiments. Eighteen patients with benign tracheal stenosis who were enrolled in 2017.06–2019.06 were enrolled, and the thickness, diameter and lumen area of the most narrow tracheal tube before and after bronchoscopy and 1 month after bronchoscopy were measured using intelligent image analysis software. Thirty male SD rats were randomly divided into three groups (Radiofrequency group, Glucocorticoids+Radiofrequency group and control group). At the 5th, and 28th day the comparisons were made on the pathological changes of rats skin,the average optical density value of TGF-β1 and the collagen density after the damage by Radiofrequency, Glucocorticoids+Radiofrequency. Results: The thickness, diameter and lumen area of the wall before and after bronchoscopy are: 3.17 ± 1.01 mm versus 1.60 ± 0.23 mm; 3.61 ± 1.23 mm versus 9.04 ± 1.00 mm; 11.19 ± 5.32 versus 67.34 ± 13.80 mm2. The scar healing was observed in Radiofrequency group, but in the Glucocorticoids+Radiofrequency group the rats’ skin were healed incompletely. The results showed significant statistical differences among three groups on TGF-β1 comparisons: 988.41 ± 111.91, 730.19 ± 109.52 and 437.37 ± 60.48 (P &lt; 0.05). The same results of comparisons on the collagen density showed: 96540.58 ± 29909.51, 43768.16 ± 6264.72 and 34521.15 ± 4636.19 (P &lt; 0.05). Conclusions: After bronchoscopy intervention, the wall thickness of benign tracheal stenosis can be reduced, and the lumen diameter and area can be increased. The results can be analyzed using CT. Locally injected Betamethasone can alleviate the cicatricial tissue hyperblastosis induced by radiofrequency.

Association between biomass formation and the prognosis of infective endocarditis due to Staphylococcus aureus

IntroductionThe aim of this study was to evaluate the association between biomass formation and the clinical characteristics and prognosis of Staphylococcus aureus infective endocarditis (IE). MethodsWe prospectively studied 209 S. aureus strains causing IE. Biomass formation was examined using the crystal violet assay and quantified spectrophotometrically. The average (SD) optical density of the biomass was compared for each clinical, microbiological (methicillin-resistance, vancomycin MIC≥1.5μg/ml) and molecular (clonal complex, agr type and agr dysfunction) variable according to their presence or absence. The primary clinical endpoints studied were in-hospital death, severe sepsis, persistent bacteraemia, symptomatic peripheral embolisms and prosthetic valve IE. ResultsMean age was 66.1 years, 61.5% of patients were male and the median age-adjusted Charlson comorbidity index was 5 points (IQR 3–8). In-hospital mortality was 37.3%. Strains belonging to CC5 and CC22 had optical biomass densities [mean (SD) 1.573 (1.14) vs 0.942 (0.98) p<0.001 and 1.720 (0.94) vs 1.028 (1.04) p=0.001, respectively]. Strains belonging to CC5 and CC22 had significantly higher optical biomass densities [1.369 (1.18) vs 0.920 (0.93) p=0.008]. No statistically significant differences were found in the clinical endpoints studied. ConclusionsHigh biomass production was associated with CC5 and CC22 but not with higher hospital mortality, septic complications, type of endocarditis, methicillin-resistance, elevated vancomycin MIC or agr dysfunction.

Intraocular Distribution and Kinetics of Intravitreally Injected Antibodies and Nanoparticles in Rabbit Eyes

PurposeTo investigate the intraocular distribution and kinetics of antibodies and nanoparticles in the experimental model.MethodsAntibodies (whole IgG 149kDa, antigen-binding fragments 48.39 kDa) and four kinds of nondegradable nanoparticles (25, 50, 200, and 250 nm) were intravitreally injected in the right eye of New Zealand white rabbits. The average optical density and concentration were used to measure intraocular distribution and kinetics.ResultsAfter intravitreal injection, antibodies were distributed throughout the vitreous humor and eliminated gradually into anterior and posterior routes. Fluorescence intensity decreased 1 day after injection and was not detected 25 days after injection. The nondegradable nanoparticles migrated posteriorly to the retina 7 days after injection onward and anteriorly to the aqueous humor from 1 hour to 1 day after injection. The fluorescence intensity of the nanoparticles was relatively stable in the vitreous humor, compared to antibodies. Nanoparticles accumulated on the internal limiting membrane of the retina with no penetration into deeper retinal tissue, whereas the smaller size 25 nm nanoparticles passed across the ciliary body and moved into choroid, retina, and suprachoroidal space. A gradual decrease of nanoparticles by their sizes in the vitreous after 30 days after injection was described as the percentage ratio: 61.1% (25 nm), 69.1% (50 nm), 78.6% (200nm), and 85.3% (250 nm).ConclusionsOur study revealed the in vivo intraocular distribution and kinetics of antibodies and nanoparticles with diverse sizes and the result might help to develop newer intraocular drugs and drug delivery systems to treat retinal diseases.Translational RelevanceThese experimental results can be valuable data for human research.

Microbial Community Analysis of Saliva and Biopsies in Patients With Oral Lichen Planus.

The specific etiology and pathogenesis of oral lichen planus (OLP) remain elusive, and microbial dysbiosis may play an important role in OLP. We evaluated the saliva and tissue bacterial community of patients with OLP and identified the colonization of bacteria in OLP tissues. The saliva (n = 60) and tissue (n = 24) samples from OLP patients and the healthy controls were characterized by 16S rDNA gene sequencing and the bacterial signals in OLP tissues were detected by fluorescence in situ hybridization (FISH) targeting the bacterial 16S rDNA gene. Results indicate that the OLP tissue microbiome was different from the microbiota of OLP saliva. Compared with the healthy controls, Capnocytophaga and Gemella were higher in OLP saliva, while Escherichia–Shigella and Megasphaera were higher in OLP tissues, whereas seven taxa, including Carnobacteriaceae, Flavobacteriaceae, and Megasphaera, were enriched in both saliva and tissues of OLP patients. Furthermore, FISH found that the average optical density (AOD) of bacteria in the lamina propria of OLP tissues was higher than that of the healthy controls, and the AOD of bacteria in OLP epithelium and lamina propria was positively correlated. These data provide a different perspective for future investigation on the OLP microbiome.

Effects of hypothermia on the repolarization duration of ventricular myocytes in rats and its mechanism

Objective: To observe the effects of hypothermia on the repolarization duration and the expression of Kir2.1 protein of ventricular myocytes in isolated rat heart and explore the role of Kir2.1 protein.Methods: Eighteen healthy adult male Sprague-Dawley rats were randomly divided into three groups (n=6 per group): Control group (C group), 35℃ group (H1 group), 32℃ group (H2 group). Langendorff isolated heart models were established. After 15 min 37℃ K-H fluid banlanced perfusion, C group continued to perfuse the K-H solution at 37℃ for 30 minutes, H1 group continued to perfuse the K-H solution at 35℃ for 30 minutes, H2 group continued to perfuse the K-H solution at 32℃ for 30 minutes. At 15 min of balanced perfusion (T1), and 30 min of continuous perfusion (T2), the heart rate,and the MAP in the three layers of the left ventricular anterior wall were recorded, the action potential duration at 50% repolarization (MAPD50), the action potential duration at 90% repolarization (MAPD90) and transmural dispersion of repolarization(TDR) were calculated. At the same time, the occurrence of arrhythmia was recorded. The expression of Kir2.1 protein was measured by Western blot. The average optical density (AOD) and the distribution of Kir2.1 protein were measured by immunohistochemistry in the ventricular tissue measured by electrophysiology. Results: Compared with T0, the heart rate was decreased, MAPD50 and MAPD90 were prolonged significantly (P＜0.05), and TDR was increased significantly (P＜0.05) in H1 group, H2 group at T1. Compared with C group, the HR was decreased, the MAPD90 was prolonged significantly (P＜0.05), TDR was increased significantly (P＜0.05),the expression and the AOD of Kir2.1 protein were decreased significantly (P＜0.05) in H1group, H2group at T1. Compared with H1 group, the heart rate of H2 group was decreased significantly (P＜0.05), MAPD50 and MAPD90 were prolonged significantly (P＜0.05), and TDR was increased significantly (P＜0.05) at T1. The distribution of Kir2.1 protein in group C was normal, while the distribution of Kir2.1 in H1 group and H2 group was disordered. Conclusion: Hypothermia prolonged the ventricular duration of repolarization and increased the dispersion of repolarization. The mechanism is related to the down-regulation the expression of Kir2.1 protein and the disorder of the distribution of Kir2.1 protein.

ОПТИМИЗАЦИЯ ПРОТОКОЛА ИММУНОФЕРМЕНТНОГО АНАЛИЗА ДЛЯ ИНДИКАЦИИ Т-2 ТОКСИНА

T-2 toxin is one of the most common and toxic trichothecene mycotoxins – secondary metabolites of molds that develop on cereals and some other crops. This article discusses the development stage of a test system for the determination of T-2 toxin based on enzyme-linked immunosorbent assay (ELISA), which uses an enzyme label – a conjugate of anti-rabbit goat antibodies with peroxidase. An important step in the creation of any ELISA-based test system is the preliminary titration of reaction components. The aim of the work was to determine the optimal concentration of the conjugate of antispecies antibodies in an indirectly competitive enzyme-linked immunosorbent enzyme immunoassay with the indication of T-2 toxin. A number of dilutions of the antivirus conjugate were examined: 1 : 1000; 1 : 2000; 1 : 3000; 1 : 4000; 1 : 5000; 1 : 7500; 1 : 10000; 1 : 12 500; 1 : 15,000; 1 : 17,500; 1 : 20,000; 1 : 30 000. In the ELISA staging protocol, calibration solutions of T-2 toxin at concentrations of 0.0 were used; 2.5; 5, 10, 20, 40, 80, 160 ng/ml and specific polyclonal rabbit antibodies to the T-2-BSA conjugate. Based on the average optical density values, calibration plots were constructed using the percentage of signal absorption from the zero standard. When evaluating the results of the study, the criterion for choosing the dilution of the conjugate of anti-species antibodies was considered the greatest, at which the best linearity of the grading plot is achieved and the level of its non-specific reaction with the zero standard would be the lowest. It was established that the optimal variants of dilutions of the conjugate of anti-species antibodies under the same experimental conditions tested were 1: 4000, 1 : 5000 and 1 : 12 500. Dose-dependent signal absorption was observed in all concentrations of anti-species antibodies. Dilutions of the conjugate of anti-species antibodies 1 : 1000–1 : 3000 and 1 : 17 500–1 : 30 000 were not taken into account, since the optical density of most wells was higher than the optimal boundaries in the first case (&gt; 3.9) and lower in the second (&lt; 0.4). Based on the foregoing, optimal dilution of the conjugate of anti-species antibodies was selected 1 : 12 500.

Simulation analysis and preparation of a high optical density laser protection filter.

High optical density (OD) filters have been widely used in space observation, optical detection, and laser protection. However, the lack of high OD value filters is restricting their application. This paper reports the formulation of a three-dimensional mesh model that can help predict the effects of pinhole defects, thickness error, and uniformity on the transmittance and OD value of optical filters. A laser protection filter (LPF) with a high OD value was prepared on fused silica using a microwave plasma-assisted pulsed DC reactive sputtering technique. The transmittance and OD value of the LPF were measured. Comparing the designed, measured, and simulated results, we found that the thickness error and uniformity of the layers mainly affected the passband transmittance of the LPF and had little effect on the OD value of the blocking band. In contrast, the pinhole defects were the main factor that decreased the OD value of the blocking band. The average OD values of the prepared LPF in the blocking bands of 527-532 and 755-833 nm were 8.832 and 10.191, respectively. By comparing the transmittance and OD value of the simulated and measured results, we found that the LPF has ${\pm 1}\% $±1% uniformity error and 0.5% pinhole ratio. Suggestions for preparing high OD optical filters are provided, and further improvements are summarized.

A Novel Smart Assistance System for Blood Vessel Approaching: A Technical Report Based on Oximetry.

In clinical practice, the catheter has to be placed at an accurate position during anesthesia administration. However, effectively guiding the catheter to the accurate position in deeper tissues can be difficult for an inexperienced practitioner. We aimed to address the current issues associated with catheter placement using a novel smart assistance system for blood vessel catheter placement. We used a hollow introducer needle embedded with dual wavelength (690 and 850 nm) optical fibers to advance the tip into the subclavian vessels in anesthetized piglets. The results showed average optical density changes, and the difference between the absorption spectra and hemoglobin concentrations of different tissue components effectively identified different tissues (p < 0.05). The radial basis function neural network (RBFNN) technique was applied to distinguish tissue components (the F-measure value and accuracy were 93.02% and 94%, respectively). Finally, animal experiments were designed to validate the performance of the proposed system. Using this system based on oximetry, we easily navigated the needle tip to the target vessel. Based on the experimental results, the proposed system could effectively distinguish different tissue layers of the animals.

Quantitative shear wave elastography in primary invasive breast cancers, based on collagen-S100A4 pathology, indicates axillary lymph node metastasis.

The purpose of this study was to evaluate the value of quantitative shear wave elastography (SWE) in indicating the axillary lymph node metastasis (LNM) of invasive breast cancers (IBCs) and to investigate if S100A4 plays a key role in promoting metastasis and increasing stiffness in IBC. The differences in SWE of 223 IBC patients were compared between the LNM+ and LNM- groups and the optimal cutoff values of SWE for diagnosing LNM were calculated. We searched the gene expression omnibus (GEO) to determine whether S100A4 was more highly expressed in IBCs that were LNM+ than in those that were LNM-. Sirius red and immunohistochemical staining were used to examine the collagen deposition and S100A4 expression of included tissue samples, and correlations of SWE and S100A4 expression with collagen deposition were analyzed. The optimal cutoff values for Emax (the maximum stiff value), Emean (the mean stiff value), and EmeanR (the ratio of Emean between mass and parenchyma) for diagnosing axillary LNM were 111.05 kPa, 79.80 kPa, and 6.89, respectively. GSE9893 exhibited more increased S100A4 expression in IBCs that were LNM+ than in those that were LNM-. Collagen volume fraction (CVF) and the average optical density of S100A4 (AODS100A4) in the LNM+ group were significantly higher than those in the LNM- group. Emax, Emean, EmeanR, and AODS100A4 were all positively correlated with CVF. SWE in primary IBC could be useful for indicating axillary LNM. S100A4 may be a factor that regulates cancer-associated collagen deposition and metastasis; however, prospective molecular biological studies are needed.

Effect of Electroacupuncture on Platelet-derived Growth Factor and the Ultrastructure of Mitochondria in Rats with Diabetic Gastroparesis

ObjectiveTo observe the effect of electroacupuncture (EA) at the pressure points Zu San Li (ST36), San Yin Jiao (SP6) and Liang Men (ST21) on platelet-derived growth factor (PDGF) and the ultrastructure of mitochondria in rats with diabetic gastroparesis (DGP). MethodsSixty Sprague Dawley (SD) rats were randomly separated into a normal control group (NC, n = 10) and a modeling group (n = 50). Rats in the modeling group received an injection of 2% streptozotocin (STZ) and a high-fat and high-glucose diet for eight weeks to establish a DGP rat model. At the same time, blood glucose and a general symptom score were recorded every week. After modeling, 30 successfully modeled rats were randomly separated into the following groups: the DGP group (n = 10), the EA group (n = 10) and the metoclopramide (MP) group (n = 10). After three weeks of intervention, the gastrointestinal propulsive rate was measured by measuring the optical density (OD). The concentration of Ca2+ was determined by fluorescence immunoassay, and levels of serum insulin (INS) and PDGF were determined by ELISA. The ultrastructure of mitochondria was observed with transmission electron microscopy. Results(1) After intervention, levels of blood glucose and the general symptom score were greatly decreased in the EA group compared to the DGP group (P < 0.01). Compared with the DGP group, the gastric emptying rate and the intestinal propulsive rate of the EA group was significantly improved (P < 0.01), and there was no statistically significant difference between the EA and the NC groups. (2) Compared with the NC group, the levels of INS in the DGP group markedly decreased (P < 0.05), but there was no significant difference of INS levels between the EA and the MP groups. (3) Compared with the DGP group, the concentration of Ca2+ in the EA and the MP groups significantly increased (P < 0.01, P < 0.05, respectively). (4) Compared with the NC group, the average OD of PDGF in the DGP group was significantly higher (P < 0.01). Compared with the DGP group, levels of PDGF in the EA group increased significantly (P < 0.01). (5) There were abundant mitochondria with a clear structure and complete cristae in the NC group. However, in the DGP group, mitochondria were severely swollen, partly vacuolated, and cristae were either fractured, absent, or shortened. In the EA group, mitochondria were slightly swollen, with clear cristae. ConclusionsElectroacupuncture at the points Zu San Li (ST36), San Yin Jiao (SP6) and Liang Men (ST21) may improve gastric motility in DGP by up-regulating the amount of PDGF and improving the ultrastructure of mitochondria.

Regulation of Vimentin on Cell Metabolic Signaling Pathway in Muscle Injury

The regulation of vimentin on cellular metabolic signaling pathway in muscle injury was analyzed experimentally. Seventy experimental mice were divided into six experimental groups and one control group according to exercise training time. Muscle injury model was established by repeated running and jumping. Vimentin expression was quantified by image analysis method, combined with pathological and ultrastructural changes of muscle tissue, and serum was detected. The regulation of vimentin metabolic signaling pathway in muscle injury cells was comprehensively analyzed by creatine kinase, cell proliferation and vimentin expression time course in skeletal muscle of mice. The results showed that the average optical density and area of vimentin positive area of mice in each experimental group were different with different exercise degrees. During the first 6–7 weeks of training, muscle injury had a process of gradual aggravation and alleviation, which indicated that the muscles showed good adaptability while the injury accumulated. The time course changes showed that there was some expression of vimentin in mice of different experimental groups after exercise. The experimental results confirmed that vimentin had a regulatory role in the metabolic signaling pathway of muscle injury cells.

The Impact of Renal Denervation on the Progression of Heart Failure in a Canine Model Induced by Right Ventricular Rapid Pacing

Heart failure (HF) has been proposed as a potential indication of renal denervation (RDN). However, the mechanisms enabling RDN to attenuate HF are not well understood, especially the central effects of RDN. The aim of this study was to decipher the mode of operation of RDN in the treatment of HF using a canine model of right ventricular rapid pacing-induced HF. Accordingly, 24 Chinese Kunming dogs were randomly grouped to receive sham procedure (sham-operated group), bilateral RDN (RDN group), rapid pacing to induce HF (HF-control group), and bilateral RDN plus rapid pacing (RDN + HF group). Echocardiography, plasma brain natriuretic peptide (BNP), and norepinephrine (NE) concentrations of randomized dogs were measured at baseline and 4 weeks after interventions, followed by histological and molecular analyses. Twenty dogs completed the research successfully and were enrolled for data analyses. Results showed that the average optical density of renal efferent and afferent nerves were significantly lower in the RDN and RDN + HF groups than in the sham-operated group, with a significant reduction of renal NE concentration. Rapid pacing in the RDN + HF and HF-control groups, compared with the sham-operated group, induced a significant increase in left ventricular end-diastolic volume and decrease in left ventricular ejection fraction and correspondingly resulted in cardiac fibrosis and dysfunction. Cardiac fibrosis evaluated by Masson’s trichrome staining and the expression of transforming growth factor-β1 (TGF-β1) were significantly higher in the HF-control group than in the sham-operated group, which were remarkably attenuated by the application of the RDN technique in the RDN + HF group. In terms of central renin–angiotensin system (RAS), the expression of angiotensin II (AngII)/angiotensin-converting enzyme (ACE)/AngII type 1 receptor (AT1R) in the hypothalamus of dogs in the HF-control group, compared with the sham-operated group, was upregulated and that of the angiotensin-(1-7) [Ang-(1-7)]/ACE2 was downregulated. Furthermore, both of them were significantly attenuated by the RDN therapy in the RDN + HF group. In conclusion, the RDN technique could damage renal nerves and suppress the cardiac remodeling procedure in canine with HF while concomitantly attenuating the overactivity of central RAS in the hypothalamus.

Experimental Study on the Effect of Aconite and Angelica sinensis on Myocardial Ischemia Rats with Yang Deficiency and Blood Stasis.

Objective To investigate the intervention effect and mechanism of Aconite and Angelica sinensis on myocardial ischemia rats with Yang deficiency and blood stasis. Methods SPF-class SD rats were randomly divided into low-dose and high-dose groups. Each group was divided into control group, model group, and drug-administered group (FZ, DG, FG; 1 : 0.5, 1 : 1, 1 : 2). A rat model was prepared by intraperitoneal injection of hydrocortisone and isoproterenol plus cold stimulation. Each group was given corresponding decoction or distilled water for 14 days. The behavioral changes of rats in each group were observed. The morphological changes of rats cardiomyocytes were observed by HE staining. The average optical density (MOD value) and percentage of positive cells of Bcl-2, Bax, and Akt were determined by immunohistochemical staining method, and PEIs were calculated. Western blot and RT-PCR were used to determine the expression of PI3K, Caspase-3, Akt protein, and gene expression. Results The compatibility of Aconite and Angelica sinensis improved the morphology of rat cardiomyocytes, increased the PEI values of Akt and Bcl-2 protein, and decreased the PEI values of Bax protein (P < 0.01). The compatibility reduced the expression of Caspase-3 protein of rat myocardium and increased the protein expression of p-Akt, PI3K, and p-PI3K (P < 0.01). The compatibility also significantly reduced the expression of Caspase-3 mRNA and increased the expression of PI3K mRNA and Akt mRNA (P < 0.05 or P < 0.01), and the effect of high-dose FG (1 : 2) group is the best. Conclusions The method of preparing a rat model of myocardial ischemia with Yang deficiency and blood stasis was feasible. The compatibility of Aconite and Angelica sinensis reduced myocardial fibrosis and inflammatory reaction, protected ischemic cardiomyocytes, and reduced myocardial injury, whose mechanism may be related to the regulation of PI3K/Akt pathway. The compatible group had better intervention effects than Aconite or Angelica sinensis alone. The best one was high-dose FG (1 : 2).

Neural Remodeling of the Left Atrium in Rats by Rosuvastatin Following Acute Myocardial Infarction.

ObjectiveThis study aims to investigate the effect of rosuvastatin on sympathetic neural remodeling of the left atrium (LA) in rats after myocardial infarction (MI).MethodsRats were randomly divided into a three groups: sham group, statin group, and MI group. The mRNA expression levels of the growth-associated protein-43 (GAP43) and nerve growth factor (NGF) were measured by RT-PCR. Immunohistochemistry was used to detect the distribution and density of GAP43- and NGF-positive nerves. The expression levels of these proteins were quantified by Western blot.ResultsCompared with the sham group, the average optical density (AOD) values of GAP43 and nerve growth factor (NGF)-positive substances in the LA in the statin and MI groups were significantly higher (P<0.01), but the AOD values in the statin group were lower than of those in the MI group (P<0.01). Furthermore, the AOD values of GAP43 and NGF positive nerves in the left stellate ganglion in the statin and MI groups were significantly higher (P<0.01), but the AOD values in the statin group were lower, when compared with the MI group (P<0.01).ConclusionRosuvastatin could effectively improve the sympathetic neural remodeling of LA in MI rats.

The role of sperm associated antigen 6 gene in morphological changes of inner ear development and signal regulation of auditory organs in mice

AimIn order to analyze the effects of the deletion of the Sperm Associated Antigen 6 (Spag6) gene on the inner ear and the auditory system of mice. MethodClustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology was utilized, and the Spag6 gene knockout mouse model was constructed. Self-breeding was carried out to obtain the F1 generation Spag6 homozygous mouse (Spag−/−), the Spag6 heterozygous mouse (Spag+/−) and the Spag6 wild mouse (Spag+/+) were obtained. Polymerase Chain Reaction (PCR) technology was used to verify the three genotypes of mice. The mean hearing threshold of Spag−/− mice, Spag+/− mice and Spag+/+ mice after different intensity of sound stimulation was detected. The inner ear cochlea tissues of Spag−/− mice and Spag+/+ mice were collected and paraffin sections were made. Morphological differences of inner ear tissues in mice were analyzed by hematoxylin – eosin (HE) staining. Immunofluorescence staining was used to analyze the number of hair cells in the inner ear of mice. Apoptosis of mouse inner ear hair cells was analyzed by TdT-mediated dUTP Nick-End Labeling (TUNEL) staining ResultIt was found that the hearing of Spag−/− mice decreased significantly compared with that of Spag+/+ mice (P < 0.01), and the hearing of Spag+/− mice decreased significantly compared with that of Spag+/− mice (P < 0.05). After HE staining and immunofluorescence staining, hair cells in the inner ear cochlea of Spag−/− mice were found to be defective. The apoptosis detection results of TUNEL staining indicated that the number of apoptosis of hair cells in inner ear cochlea of Spag−/− mice was significantly higher than that of Spag+/+ mice, and the average optical density of Corti apparatus of Spag−/− mice was significantly higher than that of Spag+/+ mice (P < 0.01). ConclusionThe results showed that the loss of the Spag6 gene accelerated the apoptosis of hair cells in the cochlear tissue of the inner ear of mice, thereby affecting the auditory system of mice. The significance of this study is to lay the foundation for future studies on the effects of Spag6 on deafness.

The effect of fine particles on the phagocytosis of alveolar macrophages potentially by Arp2/3 complex in a mouse model of chronic obstructive pulmonary disease

Objective: To investigate the mechanism of fine particulate matter (PM2.5) on the phagocytosis of alveolar macrophages (AM) in mice with chronic obstructive pulmonary disease (COPD) through actin-related protein (Arp) 2/3 complex. Methods: Forty mice were divided into healthy control(A) group, healthy PM2.5 (B) group, COPD(C) group, and COPD PM2.5(D) group according to the random number table method. A mouse model of COPD was established by cigarette smoke exposure method. PM2.5 (662 μg/m(3)) model was established by continuously inhalation for 90 days in healthy PM2.5 group and COPD PM2.5 group. Flow cytometry was used to detect the ability of AM to phagocytose fluorescein isothiocyanate-labeled E.coli (FITC-E.coli), expressed as mean fluorescence intensity (MFI) and percentage of phagocytic positive cells (phagocytosis percentage); Western blotting was used to detect AM Arp2 and F-actin content, and laser confocal microscopy for AM Arp2 and F-actin and phagocytic FITC-E.coli average optical density and colocalization of Arp2 and F-actin, while scanning electron microscopy was used to observes the morphology of AM after phagocytizing FITC-E.coli. Results: AM phagocytosis: MFI and phagocytosis percentage in the COPD group [4 656±251, (31.9±1.7)%] were lower than the healthy control group [8 657±247, (65.7±1.9)%] (both P<0.01); and healthy PM2.5 group and COPD PM2.5 group [7 653±228, (47.9±1.6)% and 3 660±237, (19.2±1.2)%] were lower than the respective control groups (all P<0.01), and the decrease in the COPD group was more pronounced. AM Arp2, F-actin content: the COPD group (0.51±0.02, 0.46±0.03) were lower than the healthy control group (0.81±0.04, 0.71±0.04, both P<0.01); the healthy PM2.5 group and the COPD PM2.5 group [(0.64±0.03, 0.56±0.04) and (0.29±0.02, 0.26±0.02)] were lower than the respective control groups (all P<0.01), and the decrease in COPD group was more significant. Arp2, F-actin, and phagocytic FITC-E.coli mean optical density values: the COPD group (33.0±2.3, 62.0±0.7, 41.0±0.4) were lower than the healthy control group (141.0±4.2, 145.0±2.9, 189.0±2.6, both P<0.01); the healthy PM2.5 group and the COPD PM2.5 group (127.0±2.8, 124.0±0.7, 154.0±0.9, and 24.0±2.4, 37.0±0.4, 29.0±0.8) were lower than the respective control groups (all P<0.01), and the decrease in the COPD group was more significant. Colocalization of AM Arp2 and F-actin: Montessori colocalization coefficient (MOC) (0.38±0.03) in the COPD group was lower than the healthy control group (0.88±0.03, P<0.01); healthy PM2.5 group and COPD PM2.5 group [(0.58±0.03) and (0.14±0.02)] were lower than the respective control groups (both P<0.01), and the decrease in COPD group was more significant. Morphology of AM phagocytosis of FITC-E.coli: AM in the healthy control group was obviously deformed, and the surface of the cell membrane was slightly wrinkled and high, and the free edge of the micro-pleated fold had a long and dense filamentous pseudopodia extension. The changes of morphology of AM in the COPD group was not obvious, the micro-wrinkles on the surface of the cell membrane were rare, and the filopodia poorly extended or even absent. The AM form of the healthy PM2.5 group changed slightly, mostly irregular circular or elliptical. The micro-wrinkles on the surface of the cell membrane were less and flat, and the filopodia protrudes short and less; the AM form of the COPD PM2.5 group was stiff, and the micro-wrinkles on the surface of the cell membrane were few and flat, no obvious filopodia or protrusions. Correlation analysis: After basal state and PM2.5 intervention, AM Arp2, F-actin content and MOC values of Arp2 and F-actin were positively correlated with MFI. Conclusions: The phagocytic function of AM in COPD mice was low, which was related to the abnormal rearrangement of cytoskeleton involved in Arp2/3 complex and F-actin. It was speculated that PM2.5 might inhibit Arp2/3 complex and F-actin. The cytoskeletal rearrangement of proteins was involved in the aggravation of AM phagocytosis in mice with COPD.

Association between biomass formation and the prognosis of infective endocarditis due to Staphylococcus aureus

IntroductionThe aim of this study was to evaluate the association between biomass formation and the clinical characteristics and prognosis of Staphylococcus aureus infective endocarditis (IE). MethodsWe prospectively studied 209 S. aureus strains causing IE. Biomass formation was examined using the crystal violet assay and quantified spectrophotometrically. The average (SD) optical density of the biomass was compared for each clinical, microbiological (methicillin-resistance, vancomycin MIC≥1.5μg/ml) and molecular (clonal complex, agr type and agr dysfunction) variable according to their presence or absence. The primary clinical endpoints studied were in-hospital death, severe sepsis, persistent bacteraemia, symptomatic peripheral embolisms and prosthetic valve IE. ResultsMean age was 66.1 years, 61.5% of patients were male and the median age-adjusted Charlson comorbidity index was 5 points (IQR 3–8). In-hospital mortality was 37.3%. Strains belonging to CC5 and CC22 had optical biomass densities [mean (SD) 1.573 (1.14) vs 0.942 (0.98) p<0.001 and 1.720 (0.94) vs 1.028 (1.04) p=0.001, respectively]. Strains belonging to CC5 and CC22 had significantly higher optical biomass densities [1.369 (1.18) vs 0.920 (0.93) p=0.008]. No statistically significant differences were found in the clinical endpoints studied. ConclusionsHigh biomass production was associated with CC5 and CC22 but not with higher hospital mortality, septic complications, type of endocarditis, methicillin-resistance, elevated vancomycin MIC or agr dysfunction.

Drivers of Clostridioides difficile hypervirulent ribotype 027 spore germination, vegetative cell growth and toxin production in vitro

ObjectivesClostridioides difficile infection (CDI) is a considerable healthcare and economic burden worldwide. Faecal microbial transplant remains the most effective treatment for CDI, but is not at the present time the recommended standard of care. We hereby investigate which factors derived from a healthy gut microbiome might constitute the colonization resistance barrier (CRB) in the gut, inhibiting CDI. MethodsCRB drivers pH, short chain fatty acid (SCFA), and oxidation–reduction potential (ORP) were investigated in vitro using C. difficile NAP1/BI/027. Readouts for inhibitory mechanisms included germination, growth, toxin production and virulence gene expression. pH ranges (3–7.6), SCFA concentrations (25–200 mM) and ORP (–300 to 200 mV) were manipulated in brain heart infusion broth cultures under anaerobic conditions to assess the inhibitory action of these mechanisms. ResultsA pH < 5.3 completely inhibited C. difficile growth to optical density (OD) 0.019 vs. 1.19 for control pH 7.5. Toxin production was reduced to 25 units vs. 3125 units for pH 7.6 (1 in 5 dilutions). Virulence gene expression reduced by 150-fold compared with pH 7.6 (p < 0.05). Germination and proliferation of spores below pH 6.13 yielded an average OD of 0.006 vs. 0.99 for control. SCFA were potent regulators of toxin production at 25 mM and above (p < 0.05). Acetate significantly inhibited toxin production to 25 units independent of OD (0.8733) vs. control (OD 0.6 and toxin titre 3125) (p < 0.05). ORP did not impact C. difficile growth. ConclusionsThis study highlights the critical role that pH has in the CRB, regulating CDI in vitro and that SCFA can regulate C. difficile function independent of pH.