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【Objective】 To investigate the expression profile of circRNA in nuclear receptor NURR1 overexpressed prostate cancer (PCa) cells, so as to provide reference for revealing the mechanism of PCa progression. 【Methods】 The expression of NURR1 in PCa was analyzed with UALCAN and TNMplot. The distinct circRNAs in NURR1 overexpressed PCa cells were screened with RNA-sequencing. The functions and signal pathways of differentially expressed circRNA molecules were analyzed with GO and KEGG. 【Results】 The circ_0000915 was significantly downregulated in DU145, LNCaP and PC3 cells. In NURR1 overexpressed DU145 cells, circ_0005991 was up-regulated, while circ_0001460 and circ_0001315 were down-regulated. In NURR1 overexpressed LNCaP cells, circ_0040729 and circ_0000722 were significantly up-regulated. In NURR1 overexpressed PC3 cells, circ_0001577, circ_0000854 and circ_0018168 were up-regulated, while circ_013035, circ_0003028, circ_0082096 and circ_0005320 were down-regulated. KEGG analysis revealed that the differentially expressed circRNAs were significantly associated with dorsal/ventral neural tube patterns, protein folding chaperones, disordered domain specific binding, positive regulation of BMP signaling pathways, and neural tube patterning functions. 【Conclusion】 CircRNAs play an important role in NURR1 mediated PCa progression, but there are certain differences among different prostate cancer cell types. The regulatory mechanism between NURR1 and circ_0000915 in the progression of PCa needs further investigation.
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BACKGROUND:Fluorosis is a disorder of enamel development caused by long-term intake of large amounts of fluoride during enamel development. OBJECTIVE:To further explore the molecular mechanism of dental fluorosis formation by screening the differentially expressed genes associated with calcium homeostasis in ameloblasts by transcriptome sequencing technology. METHODS:LS8 cells were treated with 0,0.4,0.8,1.6,3.2 and 6.4 mmol/L sodium fluoride(NaF)for 24,48 and 72 hours to observe the effects of different concentrations of NaF on the morphology,cell activity and intracellular Ca2+ concentration of LS8 cells.The differentially expressed genes were screened by transcriptome sequencing and validated. RESULTS AND CONCLUSION:After 24 hours of treatment,the cells treated with 0,0.4,and 0.8 mmol/L NaF were in good growth condition,with increased cell number and clear cell outline.When the NaF concentration was≥1.6 mmol/L,the cells were gradually shrunken and became smaller and the number of cells decreased with the increase of NaF concentration.After 48 and 72 hours of treatment,the number of cells increased in the 0,0.4 mmol/L NaF groups,while gradually decreased in the 0.8,1.6,3.2 mmol/L NaF groups,with rounded and smaller cell morphology.The cells in the 6.4 mmol/L NaF group were shrunken,rounded and suspended in the medium,with almost no adherent cells.When treated with the same concentration of NaF,LS8 cells were in optimal growth after 24 hours of treatment.Results from cell counting kit-8 assay showed that when treated with the same concentration of NaF,the cell activity decreased with the increase of treatment time;when the treatment time was the same,the cell activity decreased with the increase of NaF concentration.After 24 hours of treatment,the intracellular Ca2+ concentration increased with the increase of NaF concentration.Transcriptome sequencing analysis identified genes involved in the regulation of cellular calcium homeostasis:Hsp90b1,Canx,Calr,and Hspa5 that were significantly upregulated(P<0.05)and Cacna1a that was significantly downregulated(P<0.05).To conclude,the inhibitory effect of NaF on LS8 cell proliferation may be related to the abnormal increase in intracellular Ca2+ concentration,and the mechanism may be caused by the upregulation of the expression of protein processing and synthesis pathways Hsp90b1,Canx,Calr,and Hspa5 and the downregulation of the expression of calcium signaling pathway Cacna1a.
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Objective:To observe any effect of transcranial magnetic stimulation on brain activity, cognitive impairment and depression (PSCCID) after a stroke.Methods:Thirty patients with PSCCID after a stroke were randomly assigned into an observation or a control group, each of 15. For four weeks, both groups received basic medication to nourish the nerves, improve circulation, and anti-platelet, as well as 50mg/d of oral sertraline hydrochloride. Their rehabilitation involved 40 minutes of exercise daily, 30 minutes of assignment therapy and cognitive function training. The therapy was administered 5 days a week for the 4 weeks. In addition, the observation group received daily rTMS applied over the left dorsolateral prefrontal cortex at 10Hz and 100% of the motor threshold. Functional MRI was used before and after the treatment to assess the subjects′ cognition and depression, as well as any changes in the regional homogeneity (ReHo) and amplitude of low-frequency fluctuations (ALFFs) in regions of interest.Results:Significant improvement was observed in the subjects′ average Mini-mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA) and Hamilton Depression Scale 17-item (HAMD-17) scores, with those of the observation group significantly better than those of the control group on average. After the intervention the observation group presented several brain regions with altered ReHo and ALFF values compared with before the treatment and compared with the control group. The increases were mainly on the stimulated side while the decreases were mainly on the contralateral side. The increased ReHo in the left orbital gyrus correlated significantly with the average MMSE, MoCA and HAMD-17 scores. The increased amplitude of fluctuations in the left Heschl′s gyrus correlated significantly with the average MMSE and MoCA scores.Conclusion:rTMS can relieve depression and promote cognition after a stroke. Its effects may be associated with altered brain activity in regions related to cognitive and emotional processing.
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AIM: To analyze the strabismus surgery situation of adolescents and children in Yunnan province.METHODS: A retrospective analysis was conducted on medical records data of 3 068 adolescents and children who received strabismus surgery at Affiliated Hospital of Yunnan University from January 2017 to December 2021. The analysis included gender, constituent ratio of age, distribution of strabismus types and combination with other ocular diseases, etc.RESULTS: Among the included patients, 52.12% were males, and 47.88% were females. Preschool patients(1 to 6 years old)accounted for 32.89%, primary pupils(7 to 12 years old)accounted for 45.89% and high school students(13 to 18 years old)accounted for 21.22%. Exotropia accounted for 63.17% of the total strabismus, of which intermittent exotropia was the most common type. Esotropia accounted for 19.69%, and concomitant esotropia was the most common type. The special type of strabismus accounted for 17.14%, and A-V syndrome and dissociative vertical deviation(DVD)were the most common types. Strabismus combined with ametropia accounted for 61.02% and amblyopia accounted for 10.89%. A few patients also combined with other eye diseases.CONCLUSION: In Yunnan province, intermittent exotropia was the most common type of strabismus among adolescents and children. Some patients combined with other ocular diseases.
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Objective:To study the risk factors of failure using nasal high frequency oscillatory ventilation (nHFOV) as initial therapy in the treatment of respiratory distress syndrome (RDS) in very low birth weight infants (VLBWIs).Methods:From January 2018 to December 2021, VLBWIs with RDS initially supported by nHFOV in NICU of our hospital were retrospectively analyzed. They were assigned into success and failure groups according to the ventilation efficacy. Demographic data and clinical outcomes of the two groups were compared. Risk factors of initial nHFOV failure were analyzed using binary Logistic regression method.Results:A total of 135 infants were included, including 103 in the success group and 32 in the failure group. The initial nHFOV failure rate was 23.7%. The failure group had lower pH (7.26±0.09 vs. 7.33±0.08) and PaO 2 [61.0 (49.6, 77.2) mmHg vs. 83.6 (64.4, 99.0) mmHg] than the success group ( P<0.05) and higher PaCO 2 than the success group [49.0 (42.3, 58.1) mmHg vs. 43.4 (36.0, 50.0) mmHg] ( P<0.05). Using PaCO 2 as predictor, the area under the curve (AUC) was 0.682 (95% CI 0.575-0.788) and the cut-off value was 44.8 mmHg for nHFOV failure and the AUC was 0.716 (95% CI 0.615-0.817) and the cut-off value was 67.1 mmHg for nHFOV success. The incidences of early onset sepsis (EOS), shock within 3 d and hemodynamically significant patent ductus arteriosus (hsPDA) in the failure group were significantly higher than the success group (40.6% vs. 7.8%, 53.1% vs. 2.9%, 31.3% vs. 13.6%, P<0.05, respectively). Binary logistic regression analysis found that PaO 2<67.1 mmHg ( OR=5.458,95% CI 1.730-17.220) on the first blood gas analysis and shock within 3 d ( OR=26.585,95% CI 3.854-183.396) were independent risk factors for initial nHFOV failure ( P<0.05). Conclusions:The failure of initial nHFOV is correlated with the first blood gas parameters, EOS, hsPDA and shock within 3 d. Shock within 3 d and low PaO 2(<67.1 mmHg) were independent risk factors for initial nHFOV failure.
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Objective:To study the safety and feasibility of application of delayed umbilical cord clamping (DCC) in very preterm and extremely preterm infants.Methods:Based on the previous improvement projects of temperature management and respiratory support, we conducted a prospective study on the effect of umbilical cord clamping quality improvement project using the clinical data of very preterm and extremely preterm infants admitted to Guangdong Maternal and Child Health Hospital. The infants admitted from July to December 2020 who underwent immediate umbilical cord clamping (ICC) were included in the ICC group, and the infants admitted from January to June 2021 who underwent DCC were involved in the DCC group. The incidence of asphyxia, hypothermia, endotracheal intubation within 24 h after birth, endotracheal intubation within 72 h after birth, bronchopulmonary dysplasia and other complications, mechanical ventilation duration and total oxygen therapy duration were compared between the two groups.Results:A total of 45 cases were included in ICC group and 54 cases in DCC group. The gestational age of the two groups was (29.3±1.7) weeks and (29.6±1.4) weeks, and the birth weight was (1 250±332) g and (1 257±306) g. The differences were not statistically significant ( P>0.05). There were no significant differences between the two groups in the incidence of asphyxia, hypothermia, bronchopulmonary dysplasia and other complications related to preterm infants, tracheal intubation rates within 24 and 72 h, and the neonatal temperature at admission to NICU ( P>0.05). Conclusions:Delayed umbilical cord clamping does not increase the risks of asphyxia, hypothermia or invasive respiratory support in very preterm and extremely preterm infants.
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Objective:To investigate the clinical characteristics and short-term and long-term outcomes of preterm infants under two diagnostic criteria of bronchopulmonary dysplasia (BPD).Methods:A retrospective observational cohort study was conducted of infants admitted to the Department of Neonatology of Guangdong Women and Children's Hospital from January 2019 to June 2021. Infants who were born at <32 weeks gestational age and met the 2001 National Institute of Child Health and Human Development (NICHD) BPD definition were included. The 2019 Jenson BPD diagnostic criteria was then used to re-grade the enrolled population. Comparisons of clinical characteristics, neonatal morbidities, respiratory morbidities and growth status at follow-up between the two definition groups were conducted with t-test, nonparametric test, or Chi‐square test. Results:A total of 392 patients were included. The number of patients with BPD classified as mild, moderate, severe, and unclassifiable according to the 2001 definition was 129, 134, 114, and 15, respectively. According to the 2019 definition, there were 134 cases in the Class Ⅰ group, 89 cases in the Class Ⅱ group, 25 cases in the Class Ⅲ group, and 144 cases of non BPD. Patients with grade Ⅰ and Ⅱ BPD from the 2019 definition had an higher incidence of post-discharge home oxygen therapy, older age at discharge, and longer length of stay, compared to those with mild and moderate BPD from the 2001 definition (all P<0.05). Under both criteria, mortality increased as severity of BPD increased. Patients with grade Ⅲ BPD from the 2019 definition had higher mortality than those with severe BPD from the 2001 definition, the difference was statistically significant (48.0% vs. 14.0%, P<0.01). A total of 138 infants were followed up at 12 months of corrected age. There were no significant differences in the majority of physical development indicators and the rates of respiratory disease, readmission, and severe neurological injury, between the two definition groups at follow-up. Conclusions:The 2001 definition leads to higher BPD diagnosis rate and higher incidence of classification into the severest category within the definition. But mortality in the severe BPD group from 2001 definition was lower than that in the grade Ⅲ group from the 2019 definition. The 2019 definition is more stringent, more reasonable, more practical, and more capable of identifying patients with poor outcomes.
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Objective:To explore the relationship between the polymorphisms of AHNAK2 gene rs12882641,rs28583515 and rs2582497 loci and coronary heart disease(CHD)in the population of Xinjiang.Materials:This study used a case-control method,a total of 602 patients who were hospitalized and underwent coronary angiogra-phy(CAG)at ourheart center from Jan 2019 to Dec 2021 were selected.According toCAG results,the patients were divided into CHD group(n=301)and non-CHD group(n=301).The AHNAK2 gene rs12882641,rs28583515 and rs2582497 loci were genotyped using the improved multiple ligase detection reaction(iMLDR)technique,and the relationship between AHNAK2 gene polymorphisms and CHD was analyzed.Results:Compared with non-CHD group,there was significant rise in the distribution frequencies of AC+CC genotypes(52.8%vs.61.1%,P= 0.040)at rs2582497 locus of the AHNAK2 gene under the dominant model;there was significant reduction in distri-bution frequency of the CC genotype(65.8%vs.53.8%)at the rs28583515 locus of the AHNAK2 gene,and signif-icant rise in distribution frequencies of CT+TT(34.2%vs.46.2%)under the dominant model,TT under the re-cessive model(0.7%vs.3.0%)and CT under the additive model(33.6%vs.43.2%)in CHD group,P<0.05 or<0.01.After adjusting for confounding factors,logistic regression analysis indicated that the dominant model of the rs28583515 locus remained an independent risk factor for CHD(OR=1.509,P=0.036).Conclusion:The AH-NAK2 gene rs28583515 locus is closely related to the occurrence and development of CHD in the Xinjiang popula-tion.The dominant model of the AHNAK2 gene rs28583515 locus is an independent risk factor for CHD.
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Objective To develop a ground medical rescue platform to facilitate comfortable,safe and efficient medical rescue of astronauts at manned spaceflight landing sites.Methods A ground medical rescue platform for manned spaceflight landing site with vehicle-mounted structure was composed of a high-performance CTM-132 chassis,a rescue shelter made of aerospace aluminum plate and a hydraulic tailboard to facilitate astronauts'entry and exit.There were a variety of devices equipped in the rescue shelter for space medical monitoring and support,medical treatment and communication,and refined design was carried out for the electrical and water circuits and lighting.Results The ground medical rescue platform enabled medical monitoring and support of astronauts after returning to landing sites.Conclusion The ground medical rescue platform with the functions of medical monitoring,support and treatment meets the requirements for astronaut medical rescue at landing sites,and can be used as a supplement to the air medical rescue platform.[Chinese Medical Equipment Journal,2023,44(10):23-27]
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Aim To investigate the damage degree of doxorubicin hydrochloride( DOX )on cardiac function in rats, and to explore its possible mechanism. Methods Experiment 1: SD rats( n=48 )were randomly divided into control group( normal saline ), DOX 1 group( DOX cumulative dose 12 mg·kg-1 ;intraperitoneal injection ), DOX 2 group( 15 mg·kg-1 ;)and DOX 3 group( 18 mg·kg-1 ;). Cardiac structure and cardiac function were detected by echocardiography. B-type natriuretic peptide( BNP )was detected by ELISA. The morphological changes of myocardium were observed by Hematoxylin-eosin( HE )staining. The optimal dose group( DOX 2 group )was selected comprehensively. Experiment 2: SD rats( n=36 )were randomly divided into control group( normal saline ), DOX 2 group(15 mg·kg-1)and DOX 2+Mdivi-1 group( 15 mg·kg-1+daily abdominal injection of Mdivi-1(1 mg ·kg-1 ;)). Western blot was used to detect the protein expression of myocardial mitochondrial dynamics. Results Compared with the control group, hearts in DOX groups were enlarged and the heart function was reduced. Under the microscope, hypertrophy of cardiac cells and loose arrangement of cardiac fibers were observed in DOX group, and the higher the cumulative dose of DOX in rats, the more severe the degree of heart failure and the higher the mortality rate of rats. Compared with control group, the expression of mitochondrial dynamin-related protein 1( DRP1 )and related signaling pathway protein FUN14 domain containing 1( FUNDC1 )in DOX 2 group increased. The expression of optic atrophy 1( OPA1 )decreased, the expression of FUNDC1 and DRP1 protein decreased, and the expression of OPA1 protein was enhanced after the use of mitochondrial dynamics inhibitor(Mdivi-1). Conclusions DOX can cause chronic heart failure, and the mechanism may be related to DRP1/FUNDC1 mediated mitochondrial fission and fusion.
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Aim To observe the inhibitory effect of neferine(Nef)on the migration and invasion of non-small cell lung cancer(NSCLC)H1299 cells by blocking ROCK pathway.Methods H1299 cells were taken for in vitro culture, and treated with different concentrations of Nef.H1299 cell viability was measured by CCK-8 method to determine the dose of the experimental group.The migration and invasion abilities of H1299 cells were detected by cell scratch test and Transwell chamber test.The expression of matrix metalloproteinases MMP-2 and MMP-9 secreted from lung cancer cells was detected by enzyme linked immunosorbent assay(ELISA).The protein level of ROCK1 in H1299 cells was tested by real-time fluorescent quantitative PCR and Western blot; the binding mode and affinity between Nef and ROCK1 were stimulated by AutoDock semi flexible docking method.Results The doses of Nef in the experimental group were determined as 4, 6 and 10 μmol·L-1.These three concentrations of Nef could inhibit the migration and invasion of H1299 lung cancer cells to a certain degree in a dose-dependent manner.At the same time, Nef reduced the expression of MMP-2, MMP-9 and ROCK1 proteins related to the migration and invasion of the cancer cells.In addition, the affinity of Nef to ROCK1 was significantly higher than that of fasudil, an inhibitor of ROCK, and the binding force was stronger to A-chain of ROCK1.Conclusions As a potential natural anticancer compound, Nef can inhibit the migration and invasion of NSCLC by reducing the expression of MMP-2, MMP-9 and ROCK1 proteins related to the migration and invasion of the cancer cells.
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Objective: To study the effects on extravascular lung water of lung protective ventilation strategy applying on piglets with acute respiratory distress syndrome (ARDS) induced by paraquat (PQ) under pulse indicating continuous cardiac output (PiCCO) monitoring. Methods: The piglets models with ARDS induced by PQ were established in June 2020 and all of them were received mechanical ventilation and divided into three groups according to tidal volume (V(T)) : small V(T) group (6 ml/kg) , middle V(T) group (10 ml/kg) and large V(T) group (15 ml/kg) , there were 5 piglets in each group. The positive end expiratory pressure (PEEP) were all setup on 10 cmH(2)O. The indexes such as arterial blood gas analysis, oxygenation index (OI) , extravascular lung water index (ELWI) and pulmonary vascular permeability index (PVPI) were monitored at time of before the model was established (baseline) , time of the model was established (t(0)) and 2 h (t(2)) , 4 h (t(4)) , 6 h (t(6)) after mechanical ventilation. Lung tissue were punctured at time of baseline, t(0) and t(6) to be stained by Hematoxylin-eosin (HE) staining and pulmonary pathology were observed under light microscopy. Results: The heart rate (HR) , mean arterial pressure (MAP) and partial pressure of carbon dioxide (PaCO(2)) of all groups were higher than the base value while the pH values, partial pressure of oxygen (PaO(2)) and OI were lower than the base value when the models were established (P<0.05) . After mechanical ventilation, the HR and MAP values of all groups at t(2), t(4) and t(6) were lower than t(0) while the PaCO(2) of t(4) and t(6) were all higher than t(0), the differences were statistically significant (P<0.05) . The PaO(2) and OI of all groups showed a trend of rising at first and then decreasing after mechanical ventilation. The MAP, PaO(2), PaCO(2) and OI of the middle V(T) group and large V(T) group were apparently lower than that of the small V(T) group at t(2), t(4) and t(6) (P<0.05) . The ELWI and PVPI at t(0) of all groups were higher than that of baseline (P<0.05) . The ELWI of the small V(T) group at t(6) were lower than t(0) of the same group and t(6) of the middle V(T) group and large V(T) group (P<0.05) . HE staining showed congestion and edema of alveolar tissue, swelling of capillaries, exudation of red blood cells and widening of alveolar septum in piglets after successful modeling. And further widening of alveolar septum and rupture of alveolar septum could be seen in the lung tissues of each group at t(6), and the injury was the slightest in the small V(T) group. Conclusion: The lung protective ventilation strategy can alleviate the extravascular lung water and ARDS induced by PQ and improve oxygenation.
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Animaux , Eau extravasculaire pulmonaire , Poumon/physiologie , Paraquat/toxicité , Ventilation artificielle/effets indésirables , Syndrome de détresse respiratoire du nouveau-né/induit chimiquement , SuidaeRésumé
Objective:To study the efficacy of nasal intermittent positive pressure ventilation (NIPPV) and minimally invasive surfactant therapy (MIST) in very preterm infants with respiratory distress syndrome (RDS).Methods:From January to December 2020, very preterm infants (gestation age ≤30 weeks) with RDS born and treated in our hospital were randomly assigned into NIPPV group and nasal continuous positive airway pressure (NCPAP) group. Both groups were treated with MIST technique. The following items were compared between the two groups:adverse reactions during MIST, partial pressure of carbon dioxide (PaCO 2) at 2 h after MIST, the incidences of intubation within 72 h, two or more doses of pulmonary surfactant (PS), frequent apnea, other complications and the parameters of respiratory support treatment. Results:A total of 62 cases were included, with 32 in the NIPPV group and 30 in the NCPAP group. Compared with the NCPAP group, the NIPPV group had lower incidences of bradycardia (6.3% vs. 30.0%), decreased oxygen saturation (12.5% vs. 40.0%) and apnea (6.3% vs. 30.0%) during MIST ( P<0.05). No significant difference existed in the incidence of regurgitation ( P>0.05). PaCO 2 at 2 h after MIST [40.1(38.2,43.8)mmHg vs. 48.3(44.1,50.0)mmHg], the incidences of intubation within 72 h (6.3% vs. 30.0%), two or more doses of PS (6.3% vs. 30.0%)and frequent apnea (6.3% vs. 30.0%) in NIPPV group were lower than NCPAP group ( P<0.05). No significant differences existed between the two groups on the following items: the durations of invasive ventilation, non-invasive ventilation, oxygen therapy, the incidences of bronchopulmonary dysplasia, intraventricular hemorrhage (≥Ⅲ), periventricular leukomalacia, retinopathy of prematurity (≥Ⅱ), necrotizing enterocolitis (≥Ⅱb), nasal injury, air leak and death ( P>0.05). Conclusions:Combining NIPPV and MIST can reduce the incidence of adverse reactions during PS administration without increasing respiratory support duration and common complications in preterm infants. It is recommended for clinical use.
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Objective:To study the effects of quality improvement project of respiratory support in delivery room on the short-term clinical outcomes of very/extremely low birth weight infant (VLBWI/ELBWI).Methods:The clinical data of VLBWI/ELBWI before and after the implementation of the quality improvement project of respiratory support in delivery room in our hospital were retrospectively analyzed. The incidences of endotracheal intubation in delivery room, total endotracheal intubation within 72 h after birth, mechanical ventilation duration, total oxygen therapy duration, bronchopulmonary dysplasia (BPD) and other complications were compared between pre-improvement group (from January to October 2019) and post-improvement group (from January to December 2020).Results:A total of 85 cases were included in pre-improvement group and 85 in post-improvement group. The gestational age of the two groups were (29.2±1.4) weeks and (29.1±1.5)weeks and the birth weight were (1 180±195) g and (1 186±207) g, without significant differences ( P>0.05). After the implementation of the quality improvement project, the incidence of endotracheal intubation in the delivery room decreased from 32.9% to 2.4%, and the total incidence of endotracheal intubation within 72 h after birth decreased from 58.8% to 27.1%. The usage of pulmonary surfactant significantly decreased from 54.1% to 38.8% ( P<0.05) and the incidence of hemodynamically significant patent ductus arteriosus (hsPDA) also significantly decreased from 17.6% to 5.9% ( P<0.05).No significant differences existed among other complications ( P>0.05). Conclusions:The quality improvement project of respiratory support in the delivery room can significantly reduce the incidence of endotracheal intubation in VLBWI/ELBWI without increasing short-term adverse outcome.
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【Objective】 To investigate the effect of ssDNA aptamer of RhD blood group antigen on erythrocyte toxicity. 【Methods】 Two full-length ssDNA aptamers(82 bp) of RhD blood group antigen were obtained by gene synthesis.Five samples of whole blood with EDTA anticoagulant were collected to prepare red blood cell suspensions (4×107/mL), which were split into 10 tubes(100 μL/tube), corresponding to 5 experimental groups and 5 controls.Two monospecific full-length ssDNA sequences (100 pmol/μL, 5μL each) were added into the experimental group, while the same amount of normal saline into the control.After treatment, the experimental group and the control were incubated for 60 min at 37℃.After washing, they were suspended in LISS solution and stored at 4℃.The experimental group and the control were set according to different time point during storage (1 h, 1 d, 3 d, 10 d and 17 d), with 5 tubes in each group.For erythrocytes in LISS suspension at different storage time, Annexin V labeled with FITC was used as a probe to label the phosphatidylserine (PS) content and Fluo-4 to label Ca2+ .The eversion of PS and the change of Ca2+ concentration in red blood cells in LISS suspensions were determined by flow cytometry. 【Results】 After incubation, all groups were examined under the light microscope.No agglutination occurred in the experimental group, while agglutination occurred in the control.Flow cytometry showed that the number of Annexin V-FITC staining cells of suspended erythrocytes at the same storage time-point was similar between the experimental group and the control, with no significant differences.In the experimental group, apoptosis rate of Annexin V cells at 10-day storage(6.06±1.38) was significantly higher than that at 1-hour storage(P<0.05), so as at 17-day storage(7.77±1.23) than 1-hour, 1-day and 3-day storage(P<0.05). The apoptosis rate of Fluo-4 AM cell in suspended RBCs at the same storage time-point was similar between the two groups(P>0.05). In the experimental group, the apoptosis rate of Fluo-4 AM cell at the 3-day, 10-day and 17-day storage was 20.84±4.16, 22.35±3.37 and 27.06±2.81, respectively(P<0.05). 【Conclusion】 ssDNA aptamer was not found to have any cytotoxic effects on red blood cells, and RhD ssDNA aptamer may be used as a material for the detection and preparation of universal blood.
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OBJECTIVE@#To reveal the underlying relationships between Chinese medicine (CM) syndromes and ultrafiltration (UF) in the treatment of heart failure based on a metabonomic approach.@*METHODS@#Seventeen acute decompensated heart failure (ADHF) patients were enrolled, and their CM syndromes before and after UF were collected. In addition, their venous plasma collected before and after UF was used for liquid chromatographmass spectrometer-based metabonomic analysis. Both reversed phase liquid chromatography and hydrophilic interaction liquid chromatography were used to analyze the plasma samples. Partial least-squares to latent structure-discriminant analyses were used for data analysis.@*RESULTS@#An obvious difference was observed pre- and post-treatment. A total of 17 potential biomarkers associating with alterd syndromes with UF including hypoxanthine, 1-methylhistidine, phytosphingosine, O-decanoyl-R-carnitine, etc. were screened out, showing a significant change after UF. The major adjusted metabolic pathways were purine metabolism, histidine metabolism, leucine and isoleucine metabolism, arginine and proline metabolism, carnitine shuttle, sphingolipid metabolism and phospholipid metabolism.@*CONCLUSIONS@#Metabonomic approach is a useful tool to identify potential biomarkers of altered syndromes link to UF and could provide a theoretical basis for further research on the therapeutic mechanism of UF combined with CM.
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Objective:To investigate the effect of modified Si Junzitang on the expression of fibrous protein-5(Fibulin-5), phosphorylated protein kinase B(p-Akt )in hippocampus of rats after cerebral ischemia-reperfusion (I/R) injury and the anoikis of nerve cells. Method:The 60 male SD rats of SPF grade were randomly divided into sham operation group, model group, Edaravone group (3.2 mg·kg-1)and modified Si Junzitang high, medium and low-dose groups(19.08,9.54,4.77 g·kg-1).The middle cerebral artery occlusion (MCAO) model was established by suture method,the rats were killed 7 days later,neurological deficit score was evaluated before the death,histopathological observation was performed by hematoxylin eosin staining, apoptosis index of nerve cells was detected by TdT-mediated dUTP nick end labeling(TUNEL)staining, the expression of Fibulin-5, p-Akt and protein in ischemic hippocampus were detected by immunohistochemistry and Western blot. Result:The neurological deficit score showed that,compared with the sham operation group, the neurological deficit score of the model group was significantly increased (P<0.01), compared with model group, the neurological deficit score of Edaravone group,the high, medium, low dose groups of modified Si Junzitang were decreased (P<0.05,P<0.01). Immunohistochemical results showed that,compared with the sham operation group, the expression of Fibulin-5, p-Akt protein and the apoptosis index of nerve cells in the model group were significantly increased (P<0.05,P<0.01), compared with model group, the protein expressions of Fibulin-5 and p-Akt in Edaravone group, high, medium and low-dose groups of modified Si Junzitang were significantly increased (P<0.05,P<0.01), and the apoptosis index of nerve cells was obvious,there was a significant decrease (P<0.05,P<0.01). Western blot results showed that,compared with the sham operation group, the relative expression of Fibulin-5 and p-Akt protein in the model group was significantly down-regulated (P<0.01), compared with model group, the protein expressions of Fibulin-5 and p-Akt in the Edaravone group, the high, medium and low-dose groups of modified Si Junzitang were significantly up-regulated (P<0.05,P<0.01). Conclusion:The modified Si Junzitang may stabilize the extracellular matrix (ECM) Fibulin-5, increase the adhesion of ECM to cells and promote the expression of p-Akt protein, thus inhibiting neuronal apoptosis and protecting cerebral ischemia injury.
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Objective: To explore the clinical implication of tissue-related biomarkers in patients with acute aortic dissection (AAD). Methods: It was a cross-sectional study. Ten Stanford Type A AAD patients, who were diagnosed and surgically treated in the Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, from December 2018 to August 2019, were selected as the case group. Meanwhile, 10 patients with atherosclerotic heart disease, who underwent coronary artery bypass grafting (CABG), were selected as control group. The ascending aorta tissue specimens from patients of the two groups were collected during the operation. Four-dimensional non-standard quantitative proteomics technology (4D-LFQ) was used to detect the protein profile of ascending aorta tissue specimens of the two groups and to screen out differentially expressed proteins and analyze their biological functions. Precise quantification of the selected target proteins was achieved by parallel response monitoring (PRM). Results: A total of 3 985 proteins were identified by 4D-LFQ technology, among which 3 350 proteins could be quantified. There were 39 proteins were significantly upregulated and 47 proteins were significantly downregulated in AAD group. The results of biological function analysis showed that most of the differentially expressed proteins were located in the extracellular, and their functions were mainly involved in cell migration and proliferation, inflammatory cell activation, cell contraction, and muscle organ development. The 15 selected proteins underwent precise quantification by PRM, and the results showed that integrin α-Ⅱb (ITGA2B), integrin α-M (ITGAM), integrin β-2 (ITGB2), integrin β-3 (ITGB3) were significantly upregulated in the ascending aorta tissue of AAD patients. Conclusion: ITGA2B, ITGAM, ITGB2, and ITGB3 are highly expressed in aortic tissues of patients with AAD, which may be used as biomarkers for the diagnosis of AAD patients.
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Humains , , Aorte , Marqueurs biologiques , Pontage aortocoronarien , Études transversalesRésumé
BACKGROUND@#Non-coding RNAs have attracted considerable attention for their vital role in cancer. The purpose of this study was to determine the effects of non-coding RNAs on hepatocellular carcinoma (HCC) and reveal their regulatory mechanism in the pathophysiological process.@*METHODS@#We measured the expression of mucin 1 (MUC1) and miR-485-5p in tissues from 15 HCC patients and in liver cancer cell lines by quantitative real-time polymerase chain reaction and Western blot, screened for aberrantly expressed microRNAs (miRNAs) by miRNA microarrays. Bioinformatics tools were used to find the miRNA and circular RNA that regulated MUC1, which were validated by RNA immunoprecipitation assay and luciferase reporter assay. Cell counting kit-8, Transwell assays, and flow cytometry were used to conduct functional experiments. Proteins were examined by western blot and immunohistochemical staining assays. Significant differences between groups were estimated using the one-way analysis of variance. A P < 0.05 was considered statistically significant.@*RESULTS@#MUC1 was overexpressed in HCC tissues compared with that in paratumor tissues (normal vs. tumor, 1.007 ± 0.215 vs. 75.213 ± 18.403, t = 18.401, P < 0.001) while miR-485-5p was down-regulated (normal vs. tumor, 4.894 ± 0.684 vs. 1.586 ± 0.398, t = 16.191, P < 0.001). Inhibition of miR-485-5p promoted cell proliferation (73.33% ± 5.13% vs. 41.33% ± 3.51%, t = 8.913, P < 0.001), migration (102 ± 8 cells vs. 46 ± 8 cells, t = 8.681, P < 0.001), invasion (59 ± 7 cells vs. 28 ± 2 cells, t = 8.034, P < 0.01), and suppressed apoptosis (22.64% ± 6.97% vs. 36.33% ± 3.96%, t = 2.958, P < 0.05) of HepG2 cells with which MUC1 is knocked down. Mechanically, miR-485-5p binds to MUC1, while circHECTD1 binds to miR-485-5p, resulting in the indirect up-regulation of the MUC1 level.@*CONCLUSIONS@#Our findings reveal that circHECTD1 facilitates HCC progression by sponging miR-485-5p to up-regulate MUC1.
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Objective: To explore the predictive value of neutrophil/lymphocyte ratio (NLR) on myocardial injury in severe COVID-19 patients. Methods: In this single-center retrospective cohort study, we collected and analyzed data form 133 severe COVID-19 patients admitted to Renmin Hospital of Wuhan University (Eastern District) from January 30 to February 18, 2020. Patients were divided into myocardial injury group (n=29) and non-myocardial injury group (n=104) according the presence or absence of myocardial injury. The general information of patients was collected by electronic medical record database system. All patients were followed up for 30 days, the organ injury and/or dysfunction were monitored, the in-hospital death was compared between the two groups, and the disease progression was reevaluated and classified at 14 days after initial hospitalization. Logistic regression analysis was performed to identify risk factors of myocardial injury in severe COVID-19 patients. The ROC of NLR was calculated, and the AUC was determined to estimate the optimal cut-off value of NLR for predicting myocardial injury in severe cases of COVID-19. Results: There was statistical significance in age, respiratory frequency, systolic blood pressure, symptoms of dyspnea, previous chronic obstructive pulmonary disease, coronary heart disease history, white blood cells, neutrophils, lymphocytes, platelets, C-reactive protein, platelet counting, aspartate transaminase, albumin, total bilirubin, direct bilirubin, urea, estimated glomerular filtration rate, total cholesterol, low-density lipoprotein cholesterol, D-dimer, CD3+, CD4+, partial pressure of oxygen, partial pressure of CO2, blood oxygen saturation, other organ injury, clinical outcome and prognosis between patients with myocardial injury and without myocardial injury (all P<0.05). Multivariate logistic regression analysis showed that NLR was a risk factor for myocardial injury (OR=1.066,95%CI 1.021-1.111,P=0.033). ROC curve showed that NLR predicting AUC of myocardial injury in severe COVID-19 patients was 0.774 (95%CI 0.694-0.842), the optimal cut-off value of NLR was 5.768, with a sensitivity of 82.8%, and specificity of 69.5%. Conclusion: NLR may be used to predict myocardial injury in severe COVID-19 patients.