Objective To investigate whether miR-15b-5p can alleviate airway inflammation in asthma by negatively regulating ubiquitin specific peptidase 9X (USP9X) to down-regulate the expression of phosphatidylinositol 4, 5-diphosphate 3-kinase catalytic subunit α/AKT serine/threonine kinase 1 (PIK3CA/AKT1) pathway. Methods USP9X was predicted to be a direct target of miR-15b-5p by using an online database (miRWalk), and the luciferase reporter gene assay was performed to verify it. Co-immunoprecipitation (CO-IP) was used to verify the direct binding between USP9X and PIK3CA and the role of USP9X and its small molecule inhibitor WP1130 in the deubiquitination of PIK3CA. C57 mice were randomly divided into Control group, OVA group, OVA combined with NC group and miR-15b-5p agomir group, with 10 mice in each group. BEAS-2B cells were induced with interleukin 13 (IL-13) and treated with miR-15b-5p mimic. HE, Masson, PAS, immunohistochemistry, immunofluorescence staining, flow cytometry, Western blot and quantitative real-time PCR(qRT-PCR) were performed. Results It was found that the administration of miR-15b-5p agomir and mimic could reduce peribronchial inflammatory cells and improve airway inflammation, and miR-15b-5p could target negative regulation of USP9X. USP9X could directly bind to PIK3CA and regulate PIK3CA level in a proteasome-dependent manner, and USP9X could deubiquitinate K29-linked PIK3CA protein. Down-regulation of USP9X could increase PIK3CA ubiquitination level. WP1130, a small molecule inhibitor of USP9X, has the same effect as knockdown of USP9X, both of which could increase the ubiquitination level of PIK3CA and reduce the protein level of PIK3CA. Conclusion The miR-15b-5p/USP9X/PIK3CA/AKT1 signaling pathway may provide potential therapeutic targets for asthma.
Animals ; MicroRNAs/metabolism* ; Asthma/pathology* ; Class I Phosphatidylinositol 3-Kinases/genetics* ; Ubiquitin Thiolesterase/metabolism* ; Proto-Oncogene Proteins c-akt/genetics* ; Mice ; Signal Transduction ; Mice, Inbred C57BL ; Humans ; Inflammation/genetics* ; Cell Line ; Female ; Male

Ischemic stroke (IS) is a globally life-threatening disease. Presently, few therapeutic medicines are available for treating IS, and rt-PA is the only drug approved by the US Food and Drug Administration (FDA) in the US. In fact, many agents showing excellent neuroprotection but no blood flow-improving activity in animals have not achieved ideal clinical efficacy, while thrombolytic drugs only improving blood flow without neuroprotection have limited their wider application. To address these challenges and meet the huge unmet clinical need, we have designed and identified a novel compound AAPB with dual effects of neuroprotection and cerebral blood flow improvement. AAPB significantly reduced cerebral infarction and neural function deficit in tMCAO rats, pMCAO rats, and IS rhesus monkeys, as well as displayed exceptional safety profiles and excellent pharmacokinetic properties in rats and dogs. AAPB has now entered phase I of clinical trials fighting IS in China.

BACKGROUND:Clinical studies have shown that aerobic exercise is an important supplement to the clinical treatment of patients with pulmonary hypertension,which can alleviate the disease condition,increase exercise tolerance and improve the quality of life.However,it is not clear whether patients at different stages of pulmonary hypertension can benefit equally from exercise training. OBJECTIVE:To compare the intervention effects of early or late aerobic training on right heart failure in rats with pulmonary hypertension and explore its possible mechanism. METHODS:Sixty male Wistar rats were randomly divided into control group,model sedentary group,model early exercise group and model late exercise group,with 15 rats in each group.The model of pulmonary hypertension was established by intraperitoneal injection of monocrotaline(60 mg/kg)in the latter three groups.The model early exercise group was given 8 weeks of treadmill aerobic exercise(60%maximum running speed,60 minutes per day,5 days a week)after modeling,while the model late exercise group was trained for 6 weeks after 2 weeks of modeling.The control and model sedentary groups were fed quietly in the rat cage for 8 weeks.After training,the exercise performance,right ventricular hemodynamics,cardiopulmonary function,cardiopulmonary histopathology,reactive oxygen species level in mitochondria,activity of mitochondrial respiratory chain complex and expressions of myocardial tissue proteins were detected. RESULTS AND CONCLUSION:Compared with the model sedentary group,exercise performance and right ventricular function improved(P<0.05),myocardial collagen content,endothelin-1,tumor necrosis factor-α/interleukin-10 ratio and β-myosin heavy chain/α-myosin heavy chain ratio decreased(P<0.05),vascular endothelial growth factor and sarcoplasmic reticulum calcium-adenosine triphosphate enzyme expression increased(P<0.05),immunofluorescence intensity of mitochondrial reactive oxygen species and the protein expression of 3-nitrotyrosine decreased(P<0.05),the activities of complex I,II,IV and V increased in the model early exercise and model late exercise groups(P<0.05),but there were no significant changes in right ventricular maximum pressure,pulmonary acceleration time and pulmonary artery wall area/total vascular area ratio(P>0.05).Compared with the model late exercise group,the model early exercise group further improved exercise performance and right ventricular function,and downregulated collagen content,brain natriuretic peptide protein expression,tumor necrosis factor-α/interleukin-10 ratio and β-myosin heavy chain/α-myosin heavy chain ratio(P<0.05).To conclude,although pulmonary vascular remodeling and right ventricular overload persist in rats with pulmonary hypertension,exercise training at different stages of the disease has a cardioprotective effect.The mechanism is related to the improvement of cardiac remodeling,neurohormone system imbalance,inflammatory response and mitochondrial oxidative stress.Greater benefit is gained from initiating exercise in the early stage of the disease.

BACKGROUND:Aging is associated with increased susceptibility to cardiovascular disease,and mitochondrial dysfunction plays a key role in the pathogenesis of cardiovascular disease.Regular physical activity is beneficial to cardiovascular health and can prevent and treat chronic heart disease.However,the specific mechanism of mitochondria in the protective effect of exercise on the aging heart has not yet been clarified. OBJECTIVE:To explore the effect of aerobic exercise on cardiac pathological remodeling in aging rats and to investigate the possible mechanism of mitochondrial quality control system. METHODS:Sixty Wistar rats were randomly divided into young sedentary group(6 months old),old sedentary group(20 months old)and old exercise group(20 months old)with 20 rats in each group.Rats in the young sedentary and old sedentary groups were fed in cages for 12 weeks,while those in the old exercise group underwent moderate-intensity aerobic treadmill exercise(60%of the maximal running speed,slope 0°,60 minute per day,5 days per week)for 12 weeks.After the experiment,the heart was extracted for relevant indicator tests. RESULTS AND CONCLUSION:Cardiac morphology and myocardial histopathology:compared with the young sedentary group,the rats in the old sedentary group presented with concentric cardiac hypertrophy,myocardial fibrosis,myocardial cell apoptosis and loss,and cardiac diastolic dysfunction(P＜0.05);compared with the old sedentary group,animals in the old exercise group showed reduced myocardial fibrosis and apoptosis rates,increased cell numbers,improved cardiac function(P＜0.05),and a transition in cardiac phenotype from pathological to physiological hypertrophy.Mitochondrial function:compared with the young sedentary group,the generation rate of mitochondrial hydrogen peroxide increased(P＜0.05),respiration rate and respiratory control ratio of state 3 and state 4 decreased(P＜0.05),activities of respiratory chain complexes Ⅰ,Ⅱ and Ⅳ decreased(P＜0.05),mitochondrial calcium retention capacity decreased(P＜0.05),and mitochondrial permeability transition pore opening increased(P＜0.05)in the old sedentary group.Compared with the old sedentary group,all of the above indicators were significantly improved in the old exercise group(P＜0.05).Mitochondrial quality control:compared with the young sedentary group,mitochondrial biogenesis decreased(P＜0.05),mitophagy activity increased(P＜0.05),mitochondrial fusion reduced(P＜0.05),and fission raised(P＜0.05)in the old sedentary group;compared with the old sedentary group,mitochondrial biogenesis and mitophagy activity increased(P＜0.05),mitochondrial fusion raised(P＜0.05)and fission decreased(P＜0.05)in the old exercise group.To conclude,regular aerobic exercises exert cardioprotective effects in aging rats by regulating the mitochondrial quality control system,thus reversing pathological cardiac remodeling and improving cardiac function.

Objective To analyze the relationships of the expression of retinoid X receptor alpha (RXRA) and nuclear receptor interaction protein 1(NRIP1) in colorectal cancer tissues with the features of clinical pathology and prognosis. Methods A total of 106 specimens from colorectal cancer tissues of patients during surgery were included in colorectal cancer group (n=106), and corresponding paracancer tissue specimens were included in paracancer group (n=106). The expressions of RXRA and NRIP1 in colorectal cancer tissues were detected by immunohistochemistry. Multivariate Cox regression analysis was used to investigate th4e effects of RXRA and NRIP1 expressions on the prognosis of patients with colorectal cancer. Results The positive expression rates of RXRA and NRIP1 in colorectal cancer group were 66.04%, 69.81%, which were significantly higher than 33.96% and 30.19% in the para-cancer group (P < 0.05). The positive expression rates of RXRA and NRIP1 in patients with pathological stage Ⅲ, low differentiation, serous infiltration and lymph node metastasis were higher than those in patients with pathological stage Ⅱ, medium and high differentiation, no serous infiltration and lymph node metastasis (P < 0.05). The 3-year overall survival rates of patients with pathological stage Ⅱ, low differentiation, no serous infiltration, no lymph node metastasis, negative RXRA and NRIP1 were higher than those of patients with pathological stage Ⅲ, medium and high differentiation, serous infiltration, lymph node metastasis, positive RXRA and NRIP1 (P < 0.05). Multivariate Cox regression analysis showed serous membrane infiltration (HR=2.687; 95%CI, 1.531 to 3.156), positive RXRA (HR=3.743; 95%CI, 2.217 to 5.992), positive NRIP1 (HR=2.641; 95%CI, 1.124 to 4.757) were influencing factors for prognosis in patients with colorectal cancer (P < 0.05). Conclusion RXRA and NRIP1 are highly expressed in colorectal cancer, which are closely related to pathological stage, differentiation degree and metastasis, which can be used as biomarkers to assist in evaluating the prognosis of patients.

Objective:To study the clinical features of enlarged subarachnoid space (ESS) and its effects on brain parenchymal volume in preterm infants.Methods:From November 2014 to November 2021, a retrospective case-control study was performed on preterm infants admitted to neonatal intensive care unit of our hospital with gestational age (GA)<32 w and having brain MR imaging. At full-term of corrected GA, the superior sagittal sinus-cortical spacing (sinocortical width, SCW) was measured on brain MR imaging. The infants were assigned into ESS and non-ESS groups according to whether SCW was greater than 3.5 mm. Perinatal factors, preterm-related complications and the brain volumetric indices were compared between the two groups.Results:A total of 160 preterm infants with GA<32 w were included, 76 (47.5%) were in the ESS group, SCW:(4.48±1.47) mm, and 84 were in the non-ESS group, SCW: (2.49±0.68) mm. GA and birth weight (BW) of the ESS group were significantly smaller than the non-ESS group [(28.7±2.6) weeks vs.(29.8±2.5) weeks, (1 114±279)g vs. (1 208±290)g]( P<0.05). Small GA was an independent risk factor for the development of ESS in preterm infants with GA<32w ( OR=1.217,95% CI 1.017~1.457, P=0.032). On MR imaging, the ESS group had significantly higher total cranial cavity volume than the non-ESS group [(354.1±33.6)ml vs. (316.9±36.3) ml] ( P<0.05). No significant differences existed on head circumference, gray matter volume and white matter volume between the two groups ( P>0.05). Conclusions:ESS is common in premature infants and correlated with GA and BW. Small GA is an independent risk factor for ESS in preterm infants. ESS shows little effects on head circumference and brain parenchymal volume during early postnatal period.

Objective:To investigate the urineinterleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-33 (IL-33) and matrix metalloproteinase-9 (MMP-9) in patients with IgA nephropathy (IgAN) and their predictive value for disease progression.Methods:110 IgAN patients admitted to Zhuozhou Hospital from January 2018 to October 2021 were selected and divided into IgAN progression group (39 cases) and IgAN non progression group (71 cases) according to the progress of IgAN patients. According to the Oxford Classification Standard System (MEST-C) of IgAN, they were divided into MEST-C≥3 group (42 cases) and MEST-C<3 group (68 cases). According to the estimated glomerular filtration rate (eGFR), they were divided into eGFR≥50 ml/(min·1.73 m 2) group (group A, 63 cases) and eGFR<50 ml/(min·1.73 m 2) group (group B, 47 cases). According to the amount of urinary protein, they were divided into urinary protein≥1.5 g/24 h group (66 cases) and urinary protein<1.5 g/24 h group (44 cases). Multivariate logistic regression was used to analyze the risk factors affecting the progression of IgAN. Receiver operating characteristic (ROC) curve was drawn to analyze the value of urinary IL-6, IL-8, IL-33 and MMP-9 levels in predicting the progression of IgAN. Results:The urinary IL-6, IL-8, IL-33 and MMP-9 levels in IgAN progression group were significantly higher than those in IgAN non progression group, while the serum albumin, eGFR and complement C3 in the IgAN progression group were lower than those in the IgAN non progression group (all P<0.05). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the MEST-C≥3 group were significantly higher than those in the MEST-C<3 group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the eGFR<50 ml/(min·1.73 m 2) group were significantly higher than those in the eGFR≥50 ml/(min·1.73 m 2) group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 in the urinary protein≥1.5 g/24 h group were significantly higher than those in the urinary protein<1.5 g/24 h group (all P<0.001). Multivariate logistic regression analysis showed that the course of disease, serum albumin, eGFR, urinary protein, IL-6, IL-8, IL-33 and MMP-9 were risk factors affecting the progression of IgAN (all P<0.005). The ROC curve showed that the area under the curve (AUC) of IL-6, IL-8, IL-33 and MMP-9 in predicting the progression of IgAN was 0.956 (95% CI: 0.891-0.998), and the sensitivity and specificity were 98.4% and 86.2%, respectively. Conclusions:The elevated levels of urinary IL-6, IL-8, IL-33 and MMP-9 are closely related to the progress of IgAN, and the combination of these four indicators has a good value in predicting the progress of IgAN.

Objective:To study the changes of food characters in early swallowing recovery in patients with different laryngeal function preservation surgery.Methods:Collected patients with laryngeal cancer hypopharyngeal cancer who underwent laryngeal function preservation surgery in Lihuili Hospital of Ningbo Medical Center from January 2019 to March 2020. By fiberoptic endoscopic evaluation of swallowing (FEES) combined with Penetration and Aspiration Scale (PAS), prospectively observed the aspiration and invasion of solid, paste, fluid and other food in the early stage of trial feeding in patients with laryngeal hypopharyngeal cancer undergoing open laryngeal function preservation.Results:Among the 69 patients, 21 had vertical partial laryngectomy and 19 had partial pharyngeal partial laryngectomy. 15 days after the operation, 2 groups of patients after solid and paste food adaptability scored (1.14±0.36), (1.29±0.56) and (2.53±2.04), (2.84±2.31) points, Friedman Mtest for comparison difference had statistical significance ( Mvalues were 23.463 and 22.227, P<0.01); the liquid food for adaptability scored (2.10±1.09), (4.42±2.24) points, the pairwise comparison of liquid, solid and paste showed statistically significant differences ( tvalues were-0.976 to 1.105, P<0.05). The pairwise comparison of the adaptability of the two groups on 20 days after surgery showed no statistically significant difference ( P>0.05).Throat on the cricoid cartilage resection-ring hyoid epiglottis anastomosis (SLCP-CHEP) was 17 cases, laryngeal glottis level partial resection was 12 cases; postoperative 15 days the adaptability on the behavior of three kinds of food grade 2 groups of patients (4.65±1.90), (5.59±1.46), (6.53±1.13) points and (6.67±1.07), (4.50±2.07), (6.92±0.79) points, respectively; Minspection differences were statistically significant ( Mvalues were 29.525, 22.136, P<0.01).The pairwise comparison of solid and paste food in the two groups 20 days after the operation showed no statistically significant difference ( P>0.05), while the difference of liquid and paste was statistically significant ( tvalues were-1.375 to-0.853, P<0.05). Conclusion:In the early recovery of patients undergoing laryngeal function preservation surgery, the vertical group and the partial pharyngeal partial laryngectomy group has better adaptability to solid and paste food. The horizontal group has better adaptability to paste, the CHEP group has better adaptability to solid, and the four groups has the worst adaptability to convection, the recovery time of convective mass between the CHEP group and the horizontal group was longer. To understand the adaptability of patients with different surgical methods to food traits at the early stage of trial feeding can help to implement targeted rehabilitation programs, carry out progressive diet training, and reduce the complications of misinvasion, aspiration, aspiration pneumonia and other complications in the recovery cycle.

There remains a significant gap in our quantitative understanding of crosstalk between apoptosis and necroptosis pathways. By employing the SWATH-MS technique, we quantified absolute amounts of up to thousands of proteins in dynamic assembling/de-assembling of TNF signaling complexes. Combining SWATH-MS-based network modeling and experimental validation, we found that when RIP1 level is below ~1000 molecules/cell (mpc), the cell solely undergoes TRADD-dependent apoptosis. When RIP1 is above ~1000 mpc, pro-caspase-8 and RIP3 are recruited to necrosome respectively with linear and nonlinear dependence on RIP1 amount, which well explains the co-occurrence of apoptosis and necroptosis and the paradoxical observations that RIP1 is required for necroptosis but its increase down-regulates necroptosis. Higher amount of RIP1 (>~46,000 mpc) suppresses apoptosis, leading to necroptosis alone. The relation between RIP1 level and occurrence of necroptosis or total cell death is biphasic. Our study provides a resource for encoding the complexity of TNF signaling and a quantitative picture how distinct dynamic interplay among proteins function as basis sets in signaling complexes, enabling RIP1 to play diverse roles in governing cell fate decisions.
Animals ; Apoptosis ; Caspase 8/metabolism* ; GTPase-Activating Proteins/metabolism* ; HEK293 Cells ; Humans ; Mice ; Mice, Knockout ; Necroptosis ; Receptor-Interacting Protein Serine-Threonine Kinases/metabolism*

Objective:To investigate the efficacy and safety of different dosage regimens of levosimendan in elderly patients with severe heart failure.Methods:Thirty-two patients 75 years or older were randomly divided into a loading dose group (16 cases) in which levosimendan was maintained at 0.1 μg·kg -1·min -1 for 24 h after loaded with 6 μg/kg, and a maintenance dose group (16 cases) with same schedule without loading dose. The amino-terminal brain natriuretic peptide (NT-proBNP) before and after treatment was detected. Left ventricular ejection fraction (LVEF), stroke volume (SV), stroke volume index (SVI) by echocardiograph were monitored. Adverse events, the length of stay in ICU and 28-day mortality were recorded. Results:The NT-proBNP level in loading group after treatment was 1 950 (922,6 481)ng/L, which was improved than that before treatment [4 018(2 716,9 637)ng/L, P<0.05]. The result was similar in maintenance group [1 390 (599,3 297)ng/L vs. 4 576 (2 681,10 682)ng/L, P<0.05]. LVEF in loading group before and after treatment was (39.4±8.8) % vs. (48.9±9.2) % respectively, while in maintenance group it was (40.4±8.8) % vs. (48.7±12.0) % (both P<0.05). SV were also improved after treatment in both groups compared with baseline levels ( P<0.05). NT-proBNP started to decline on day 3 in the loading group, while on day 7 in the maintenance group. SVI recovered on day 14 in the loading group [ (29.4±6.5) ml/m 2 vs. (27.3±6.7) ml/m 2, P<0.05], while it did not change much in the maintenance group. There was no significant differences as to the length of stay in ICU [ (11.1±4.4) d in loading group vs. (9.6±3.5) d in maintenance group] and 28-day mortality rates were comparable (2/16 in loading group vs. 1/16 in maintenance group) . The adverse events were 7 vs. 2 cases in loading group and maintenance group respectively, which were mild and all alleviated. Conclusion:The application of levosimendan only with maintenance dose improves cardiac function in very elderly patients with severe heart failure. Adverse events are mild and manageable.