Effect of pulmonary surfactant combined with budesonide in improving oxygenation and clinical outcomes in neonatal acute respiratory distress syndrome / 解放军医学杂志

Objective To explore the role of pulmonary surfactant(PS)combined with budesonide in improving oxygenation and clinical outcomes of neonatal acute respiratory distress syndrome(ARDS).Methods The present study is a historically controlled trial.Infants with ARDS requiring mechanical ventilation and PS replacement therapy were collected from the neonatal unit of Southwest Medical University.Those from January 2022 to November 2022 were set as intervention group(PS+ budesonid,n=35),treated with intratracheal instillation of a mixed suspension of budesonide(0.25 mg/kg)and PS(200 mg/kg),and continuous budesonide nebulization(0.25 mg/kg,twice per day)until withdrawal,then compared with a historical cohort,who just received intratracheal instillation of PS(200 mg/kg)(January 2020-December 2021,PS group,n=35).Baseline data such as gender,mode of delivery,1 min and 5 min Apgar score,birth weight,gestational age,time of onset,and cause of onset were recorded in both groups.The oxygenation and clinical outcomes of infants were compared between the two groups,including:(1)Arterial blood gas analysis indicators,such as partial pressure of oxygen(PaO2)and oxygenation index(OI)before treatment and at 6,12 and 24 hours of treatment;(2)Clinical observation and evaluation indicators,such as the time to withdrawal,duration of oxygen supplementation,length of stay,improvement of the radiological images of the lungs at 72 h of treatment,and repeated PS use;(3)Blood chemistry indicators,such as white blood cell(WBC),neutrocyte(NEU),procalcitonin(PCT)before treatment and at 3 and 7 days of treatment;and(4)Observation indicators of complications,weight growth,and mortality outcomes,such as the incidences of intracranial hemorrhage,gastrointestinal hemorrhage,neonatal necrotizing enterocolitis(NEC),and hyperglycemia,weight growth,and fatality rate.Results The differences in baseline data between the two groups were not statistically different(P＞0.05).The levels of PaO2 of the two groups were increased after treatment for different time periods,while the levels of OI were decreased(P＜0.001),and the levels of above indexes changed more significantly in PS+budesonide group than those in PS group(P＜0.05).The time to withdrawal,duration of oxygen supplementation,and length of stay in PS+budesonide group were shorter than those in PS group;the radiological images of the lungs showed that the pulmonary inflammation absorption was significantly better in PS+ budesonide group than that in PS group,while no significant difference between the two groups of infants with repeated PS use.The NEU was significantly higher in PS+budesonide group than in PS group at 3 d and 7 d of treatment(P＜0.001);and at 3 days of treatment,the PCT levels were significantly lower in PS+budesonide group than that in PS group(P＜0.05).The incidences of intracranial hemorrhage,gastrointestinal hemorrhage,NEC,hyperglycemia,weight growth,and fatality rate were not significantly different between the two groups(P＞0.05).Conclusion The use of budesonide in addition to surfactant may improve the oxygenation of neonates with ARDS,improve the inflammatory infiltrates in lungs,shorten the duration of mechanical ventilation and oxygen supplementation,and without short-term complications associated with budesonide use.

Automated Framework and Visualization-based Analysis of Concept Evolution of Spleen and Stomach Diseases and Syndromes in Traditional Chinese Medicine / 中国实验方剂学杂志

ObjectiveTo collect and screen records concerning the spleen and stomach diseases and syndromes in ancient books of traditional Chinese medicine （TCM） using an automated framework and to systematically explore the concept evolution of spleen and stomach diseases and syndromes using the visualization method. MethodA total of 1 224 kinds of ancient book data in the Ancient Books of Traditional Chinese Medicine Database （V2.1） were analyzed using the automated testing tool Selenium WebDriver with the Python 3.8 programming language and the etree library of Lxml for automatic collection and statistics of the "book title" "author" "classification" "dynasty" "completion time"， and "informative abstract". After being checked and collated， the collected data were visually analyzed with Tableau （V2020.1.3） for figuring out the concept evolution of spleen and stomach diseases and syndromes in the past dynasties from the perspectives of symptoms and signs， etiology and pathogenesis， principle-method-recipe-medicinal， and prognosis. ResultA total of 7 203 clauses were automatically collected from 989 ancient books. It was found that in the pre-Qin period， there were few ancient books related to the spleen and stomach diseases and syndromes， and the understandings were confined to the superficial symptoms or signs and the basic etiology and pathogenesis. From the Han to Sui and Tang dynasties， the related concepts gradually increased and the descriptions about the manifestations are more detailed than those in previous dynasties. The etiology， diagnosis， and treatment system of the spleen and stomach diseases and syndromes were further perfected. In the Song， Jin， and Yuan dynasties， such concepts as independent signs，symptoms， as well as nature and location of spleen and stomach diseases and syndromes were enriched. In the Ming and Qing dynasties， a TCM syndrome differentiation and treatment system for spleen and stomach diseases and syndromes was formed， and the related concepts were gradually simplified and unified. ConclusionThe concepts of spleen and stomach diseases and syndromes have undergone an evolution from simplicity to complexity and then back to simplicity. There are numerous ancient books discussing the concepts of spleen and stomach diseases and syndromes， exhibiting a fluctuating yet rising trend with time. The automated framework enables the construction of a lightweight database of spleen and stomach diseases and syndromes. Based on data visualization， the concept evolution of the spleen and stomach diseases and syndromes from ancient times to the present has been efficiently uncovered， which is conducive to tracing the origin and development of spleen and stomach diseases and syndromes in TCM. This has provided reference for related research of spleen and stomach diseases in modern Chinese medicine.

Automatic Framework Acquisition and Visualization Analysis of Ancient Chinese Medicine Literature Relating to Tumor / 中国实验方剂学杂志

Objective:To obtain ancient traditional Chinese medicine（TCM）literatures relating to tumor and visual analysis by an automatic framework tool， in order to systematically sort out the development of ancient Chinese medicine oncology. Method:Based on the database platform of ancient TCM books，names of tumor-related diseases in ancient TCM books were retrieved by Selenium WebDriver， an automation framework tool under Python 3.8. Lxml's etree library was used to parse the data. Statistics was made for "classification"， "authors"， "completion time" and "summary" of relevant ancient books automatically. After the data was checked and processed， Tableau 2019.2 software was used for data visualization analysis. And ancient Chinese medicine literatures relating to tumor were consulted at the database manually，with the dynasties as the clue，and the symptoms，etiology，pathogenesis and prognosis as the emphasis，this paper explores the development process of TCM oncology. Result:A total of 774 349 bytes of text data of 1 128 entries in 242 ancient books were included automatically. According to the findings， there were simple classification and time distribution of tumor diseases in ancient TCM books in the pre-Qin period， with a simple view on the pathogenesis of tumor diseases. From the Han dynasty to the Tang dynasty， the number of relevant literature records and the types and disease names had gradually increased，which further enriched the cognition of tumor nature，signs，classification methods，differential diagnosis；in Song and Ming dynasties，the proportion of Chinese prescription books and surgery books had increased gradually，with the largest number of abdominal organ tumor names among all dynasties；from Qing dynasty to the Republic of China，literatures relating to tumor name and classification were the most improved，and then the TCM tumor syndrome differentiation and treatment system had been formed. Conclusion:It was found that TCM oncology originated in the pre-Qin dynasty，and was improved in the Han and Tang dynasties， mature in the Song and Ming dynasties and completed in the Qing dynasty and the Republic of China. The data visualization method with integrated automation framework and parsing tools is helpful to analyze the subdivision characteristics of ancient TCM literatures，which is convenient，efficient and innovative，in the expectation to provide a classic reference for contemporary TCM studies.

Effect of hsa_circ_0001613 on Zika virus replication: Preliminary study / 中国输血杂志

【Objective】 To investigate the effect of transfusion-transmitted Zika virus (ZIKV) on the expression of non-coding circular RNA (hsa_circ_0001613) and the role of hsa_circ_0001613 in Zika virus replication. 【Methods】 Human adenocarcinomic alveolar basal epithelial cells (A549) were seeded on a 12-well plate at 1.8×105/ well and infected with ZIKV at 0.05 MOI. The Total RNAs were isolated every day for 5 days after infection, and the relative expression level of hsa_circ_0001613 was detected by qRT-PCR. In addition, 10nM siRNA-hsa_circ_0001613 was transfected into 2×105/ well A549 cells to specifically knock down the expression level of hsa_circ_0001613. 24h later, the cells were infected with ZIKV (MOI=0.05). Total RNAs were isolated at day 1-5 post-infection, proteins were extracted 96h post-infection. ZIKV replication, relative host antiviral gene expression, and interferon stimulated response element (ISRE) activity were tested using qRT-PCR, western blot and dual luciferase assay, respectively. 【Results】 The relative expression of hsa_circ_0001613 decreased significantly after 1-5 days of ZIKV infection. Knockdown of hsa_circ_0001613 inhibited ZIKV replication. Meanwhile, hsa_circ_0001613 knockdown significantly upregulated IFN-α/β and its downstream interferon-stimulated genes (ISGs) expression, also increased ISRE activity. 【Conclusion】 ZIKV infection significantly suppressed hsa_circ_0001613 expression in A4549 cells. Preliminary study indicated that hsa_circ_0001613 knockdown inhibited ZIKV replication possibly through activating type-Ⅰ IFN signaling pathway as showed by increased ISGs expression and ISRE activity.

Dynamic observation of pulmonary function by plethysmography in preterm infants with bronchopulmonary dysplasia / 中国当代儿科杂志

OBJECTIVE@#To study the effect of bronchopulmonary dysplasia (BPD) on lung function in preterm infants.@*METHODS@#According to the presence/absence or the severity of BPD, 72 preterm infants were divided into non-BPD group (n=44), mild BPD group (n=15) and moderate BPD group (n=13). Lung function was assessed by plethysmography on days 7, 14 and 28 after birth.@*RESULTS@#The preterm infants in the three groups had gradual increases in tidal volume per kilogram (TV/kg), functional residual capacity (FRC), ratio of time to peak tidal expiratory flow to total expiratory time (%T-PF) and ratio of volume to peak tidal expiratory flow to total expiratory volume (%V-PF) on days 7, 14 and 28 after birth, while there were gradual reductions in effective airway resistance per kilogram (Reff/kg) and respiratory rate (RR) (P<0.05). Compared with the non-BPD group on days 7, 14 and 28 after birth, the mild and moderate BPD groups had significantly lower TV/kg, FRC, %T-PF, and %V-PF and significantly higher Reff/kg and RR (P<0.05). On day 7 after birth, the moderate BPD group had significantly higher airway resistance, Reff/kg and FRC/kg than the mild BPD group (P<0.05).@*CONCLUSIONS@#There is a certain degree of pulmonary function impairment in preterm infants with BPD. Dynamic monitoring of lung function by plethysmography is useful for assessing lung development in the neonatal period in these infants.

Kaempferol alleviates ox-LDL-mediated endothelial cell injury via regulating AMPK/Nrf2/HO-1 signaling pathway / 中国免疫学杂志

Objective:To investigate the effects of kaempferol on the oxidized low density lipoprotein(ox-LDL)-induced injury in human umbilical vein endothelial cell (HUVECs) and to explore its underlying molecular mechanism.Methods: HUVECs were randomly divided into 6 groups:control group,ox-LDL group,Kaempferol+ox-LDL group,Kaempferol+ox-LDL+Compound C group, Kaempferol+ox-LDL+si-Nrf2 group and Kaempferol+ox-LDL+si-HO-1 group.The cell activity was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.The protein expression were measured by Western blot.The ROS levels were evaluated by flow cytometry.Results: Compared with control group,ox-LDL treatment decreased the cell viability,increased the cell apoptosis,up-regulated the protein levels of cleaved-caspase 3 and down-regulated the Bcl-2 level,elevated the expression of TNF-α,IL-1β,IL-6, vascular cell adhesion molecule (VCAM1),intercellular adhesion molecule (ICAM1) and selectin E (E-selectin),promoted the reactive oxygen species (ROS) generation and reduced the superoxide dismutase(SOD) levels,and decreased the protein levels of p-AMPK,Nrf2 and HO-1.Compared with the ox-LDL group,kaempferol treatment reversed the ox-LDL-induced cell activity inhibition, apoptosis and oxidative stress damage,and increased the protein levels of p-AMPK,Nrf2 and HO-1.Compound C,si-Nrf2 and si-HO-1 inhibited AMPK/Nrf2/HO-1 signaling pathway,elevated the production of ROS and thus inhibiting the protective effects of kaempferol on ox-LDL-treated HUVECs.Conclusion:Kaempferol alleviates ox-LDL-induced endothelial cell injury,which may be related with the activation of AMPK/Nrf2/ HO-1 signaling pathway.

Effect of Mild Hypothermia Therapy on Neonatal Bilirubin Encephalopathy:Evaluated with 18F-fluorodeoxyglu-cose Positron Emission Tomography/CT and Amplitude Integrated Electroencephalogram / 中国康复理论与实践

Objective To investigate the clinical effect of mild hypothermia on neonatal bilirubin encephalopathy, and the value of 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)/CT and amplitude integrated electroencephalogram (aEEG) for diagnosis and evaluation of curative effect. Methods From May, 2013 to December, 2014, 29 newborns with bilirubin encephalopathy were divided into conventional group (n=15) and mild hypothermia group (n=14). The conventional group received conventional therapy, and the other group received mild hypothermia in addition. The aEEG and neuron-specific enolase (NSE) were measured before and after treatment, as well as the glucose metabolism rate with 18F-FDG PET/CT after treatment. Results The NSE was lower after treatment in both groups (t>9.670, P2.943, P0.640, P<0.05). Conclusion Mild hypothermia therapy could further promote the energy metabolism of brain cells in neonatal bilirubin encephalopathy. 18F-FDG PET/CT and aEEG can be used for early diagnosis and therapeutic evaluation.

Meta-analysis of the efficacy and safety of inhaled corticosteroids for preventing chronic lung disease in preterm infants / 临床儿科杂志

Objective To evaluate the efficacy and safety of inhaled corticosteroids for preventing chronic lung disease (CLD) in preterm infants. Methods PubMed, EMBASE, CENTRAL, the ISI Web of Knowledge databases, CBM, CNKI, VIP and Wanfang Data were searched for the period up to Oct. 2016. All randomized controlled trials (RCTs) about inhaled corticosteroids for preventing CLD in preterm infants were collected. The RCTs had been screened, data were extracted and assessed. The mata-analysis was performed by RevMan 5.3 software. Result A total of 12 RCTs were included (a total of 2051 preterm neonates). Compared with control group, in 28 day old group, the incidence of CLD was not significantly different between experimental and control groups (RR=0.87, 95%CI:0.74-1.03, P=0.11) and (RR=1.19, 95%CI:0.59-2.43, P=0.63) and no significant difference among subgroups budesonide (α), beclomethasone (β), fluticasone (γ) (RR=0.89, 95%CI:0.69-1.14, P=0.35), (RR=0.86, 95%CI:0.69-1.08, P=0.19) and (RR=0.91, 95%CI:0.60-1.38, P=0.19). In 36 wk postmenstrual age group,the incidence of CLD was decreased in experimental group and in subgroups inhalation (A), Intratracheal administration (B), α, γ (RR=0.70, 95%CI: 0.61-0.80, P<0.00001), (RR=0.74, 95%CI: 0.63-0.87, P=0.0003), (RR=0.57, 95%CI: 0.43-0.76, P=0.0002), (RR=0.67, 95%CI: 0.57-0.78, P<0.00001) and (RR=0.58, 95%CI: 0.36-0.94, P=0.03); but it is not significantly different in subgroup β(RR=0.98, 95%CI: 0.69-1.39, P=0.90); There was no difference in the motality in experimental and subgroups A ,B, α, β , γ (RR=1.07, 95%CI:0.86-1.33, P=0.55), (RR=1.24, 95%CI: 0.97-1.59, P=0.09), (RR=0.67, 95%CI: 0.43-1.03, P=0.07), (RR=1.04, 95%CI: 0.81-1.33, P=0.78), (RR=1.47, 95%CI: 0.79-2.74, P=0.22) and (RR=0.91, 95%CI: 0.47-1.74, P=0.77). No clinically significant adverse effects were observed during the study. Conclusions This updated review indicated that early administration of inhaled steroids to very low birth weight preterm neonates was effective in reducing the incidence of CLD. There was no statistically significant effect of inhaled steroids on motality, and there was no significant correlation between the mode of administration and the type of drug delivery, It is recommended to observe the 36 week gestational age as the outcome index. More and larger randomised placebo-controlled trials including long-term follow up are needed to establish the efficacy and safety of inhalation corticosteroids.

Resveratrol increases sirtuin 1 expression in peripheral blood mononuclear cells of premature infants and inhibits the oxidative stress induced by hyperoxia in vivo / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the effect of resveratrol on the levels of sirtuin 1 (SIRT1) and reactive oxygen species (ROS) in peripheral blood mononuclear cells (PBMCs) of premature infants exposed to hyperoxia.</p><p><b>METHODS</b>Peripheral blood and isolated PBMCs from premature infants (gestational age<32 weeks) without oxygen supplement were collected and were randomly assigned into four groups: control, air+resveratrol, hyperoxia, and hyperoxia+resveratrol. The PBMCs were cultured in vitro for 48 hours, then the ROS content in PBMCs was measured by laser scanning confocal microscopy. Malondialdehyde (MDA) content in the medium was measured by the whole spectrum spectrophotometer. SIRT1 positioning was assessed by immunofluorescence. SIRT1 expression levels in PBMCs were measured by Western bolt.</p><p><b>RESULTS</b>Compared with the control group, the level of SIRT1 in the air+resveratrol group increased significantly (P<0.05). The levels of ROS and MDA and the SIRT1 transposition rate in the hyperoxia group increased significantly, while the expression level of SIRT1 decreased significantly compared with the control group (P<0.05). The levels of ROS and MDA and the SIRT1 transposition rate decreased significantly (P<0.05), and the expression level of SIRT1 increased significantly in the hyperoxia+resveratrol group (P<0.05).</p><p><b>CONCLUSIONS</b>Resveratrol can increase SIRT1 expression in PBMCs and inhibit SIRT1 shuttle from nucleus to cytoplasm in order to increase the ability of antioxidative stress in premature infants exposed to hyperoxia, thereby reducing the oxidative stress injury in premature infants.</p>

Changes in serum cortisol and adrenocorticotropic hormone in children with sepsis shock and their clinical significance / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes in serum levels of cortisol and adrenocorticotropic hormone (ACTH) in children with septic shock (SS) and to explore their relationship with the disease severity and prognosis.</p><p><b>METHODS</b>Twenty-five children with decompensated SS and 24 children with early SS were enrolled. Serum cortisol and ACTH levels were determined on admission and days 3 and 8 after admission. Twenty-five healthy children were used as the control group. The children with decompensated SS were further divided into death group (n=5) and survival group (n=20) based on their clinical outcome.</p><p><b>RESULTS</b>On admission, the decompensated SS and early SS groups had significantly higher serum cortisol and ACTH levels than the control group (P<0.05), and the decompensated SS group had significantly higher serum cortisol and ACTH levels than the early SS group. On day 3 after admission, the decompensated SS group had significantly higher serum cortisol and ACTH levels than the early SS and control groups (P<0.05), and the early SS group had a significantly higher serum ACTH level than the control group (P<0.05). Among the children with decompensated SS, the death group had significantly higher serum cortisol and ACTH levels than the survival group on admission (P<0.01); on day 3 after admission, the death group still had a significantly higher serum cortisol level than the survival group (P<0.01).</p><p><b>CONCLUSIONS</b>Children with SS have increased serum cortisol and ACTH levels, which are associated with the disease severity. A persistent high serum cortisol level indicates a poor prognosis. Dynamic monitoring of serum cortisol and ACTH levels in children with SS is of great significance in evaluating the disease severity and prognosis.</p>

Effect of Budesonide combined with Poractant Alfa preventing bronchopulmonary dysplasia in preterm infants / 中华实用儿科临床杂志

Objective To evaluate the effect of Budesonide combined with Poractant Alfa(pulmonary surfactants,PS) on preventing the bronchopulmonary dysplasia (BPD) in preterm infants.Methods One hundred and twenty preterm infants 6 hours after birth(gestational ages≤32 weeks and birth weights ≤1500 g)admitted to the Department of Newborn Medicine,the Affiliated Hospital of Southeast Medical University from October 2013 to February 2015 were randomly divided into 4 group(30 cases in each group).Group A was a control group,group B was neonatal respiratory distress syndrome(NRDS) group,group C was NRDS with PS group,and group D was NRDS with PS and Budesonide group.Thirty-two-week preterms without other diseases and without uptaking oxygen within 48 h were assigned as group A.Group B were treated by continuous uptaking oxygen with continuous positive airway pressure(CPAP) (oxygen uptaken lasting more than 48 h and oxygen concentrations more than 40％).Group C were treated with 100 mg/kg PS within 48 h on the basis of group B.Group D were treated with 0.25 mg/kg Budesonide suspension for inhalation on the basis of group C.The pH value,partial pressure of oxygen [pa(O2)],partial pressure of carbon dioxide [pa(CO2)] in the blood gas analysis were all detected in all groups before treatment,1,6,12,24 and 48 hours after using drug,respectively.All groups were also observed for whether to use respirator assisted ventilation,the duration of high oxygen use,the total time of uptaking oxygen,the rate of using PS again,the rate of BPD,the total hospitalization days and the adverse effects.The adverse effects included high blood pressure,high blood sugar,necrotizing enterocolitis and the incidence of nosocomial infection.Results Compared with group B,the pH value at 1 and 6 hours after using drugs,the pa(O2) and pa(CO2) at 1,6 and 12 hours after using drugs were improved obviously in the group C,and the differences were statistically significant (all P＜0.01).Compared with group B,the above indicators were improved more obvious in group D,and the differences were statistically significant (all P＜0.01).Moreover,compared with the group B,the oxygen inhalation duration,the rate of having a respirator assisted ventilation and using PS again,and the incidence of BPD were obviously decreased in other groups,the differences were statistically significant (all P＜0.05).The incidence of BPD in group D was less than that of group C,and the differences were statistically significant (3.33％ vs 10.00％,x2=4.00,P=0.046).The total oxygen time and the rate of adverse effects of each group were similar.The differences were not statistically significant (all P＞0.05).Conclusions Budesonide combined with Poractant Alfa can prevent BPD in preterm infants.Its effect is better than the simple use of Poractant Alfa,and the rate of adverse effects are not increased significantly.

Prophylactic probiotics for preventing necrotizing enterocolitis and reducing mortality in very low birth weight infants: a Meta analysis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To systematically evaluate the efficacy and safety of probiotic supplementation for preventing necrotizing enterocolitis (NEC) and reducing mortality in preterm very low birth weight (VLBW) infants.</p><p><b>METHODS</b>The randomized controlled trials (RCTs) about probiotics for preventing NEC in preterm neonates were searched in PubMed, EMbase, Cochrane Central Register of Controlled Trials (CENTRAL), the ISI Web of Knowledge databases, China Biology Medicine disc (CBM), China National Knowledge Infrastructure (CNKI), Weipu and Wanfang Data from their establishment to March 2014. The Cochrane Collaboration's RevMan 5.1 Software was used for a Meta analysis.</p><p><b>RESULTS</b>A total of 21 RCTs involving 4 607 preterm VLBW infants were eligible for inclusion in the Meta analysis. The Meta analysis showed that probiotic supplement was associated with a significantly decreased risk of NEC in preterm VLBW infants (RR=0.47; 95%CI: 0.35-0.62; P<0.001). Risk of mortality was also significantly reduced in the probiotic group (RR=0.63; 95%CI: 0.51-0.78; P<0.01). Probiotic supplement did not decrease the risk for sepsis (RR=0.87; 95%CI: 0.72-1.06; P=0.17) and NEC related mortality (RR=0.68; 95%CI: 0.31-1.48, P=0.33).</p><p><b>CONCLUSIONS</b>The results confirm that probiotic supplement can reduce risk of NEC and mortality in preterm VLBW infants. However, the long-term effects and safety of probiotics need to be assessed in large trials.</p>

Mechanism for p47phox-induced reactive oxygen species increasing after oxygen therapy in premature infants / 中华实用儿科临床杂志

Objective To explore the mechanism for the increase in reactive oxygen species regulated by p47phox of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit in peripheral blood mononuclear cells (PBMCs) after oxygen therapy in premature infants.Methods According to different volume fractions of oxygen,premature infants less than 32 weeks were divided into 3 groups:fractional concentration of inspired oxygen (FiO2) ＜ 30％ was low concentration oxygen group,FiO2 between 30％ and 40％ as middle concentration oxygen group,and FiO2 ＞ 40％ as high concentration oxygen group.Premature infants less than 32 weeks without oxygen was control group.After 48 h,3 mL blood was collected via radial artery from each group,PBMCs and serum were separated.Then intracellular reactive oxygen species (ROS) by confocal laser scanning microscopy,malondialdehyde (MDA) within serum by thiobarbituric acid colorimetric,and the location and activation rate of p47phox through immunofluorescence.Results After premature infants were exposed to oxygen,as the oxygen volume fraction was increasing,ROS and MDA gradually rised.More PBMCs with p47phox translocated to membrane,then the translocation rate of p47phox also increased.Compared with the control group,ROS were significantly higher(q =4.48,6.5,16.22,all P ＜ 0.05) among the other 3 groups ; MDA significantly increased as well(q =5.08,8.22,12.76,all P ＜ 0.05) ; the activation rate of p47phox also had significant differences (x2 =134.008,P ＜ 0.05);compared with the middle concentration oxygen group,the high concentration oxygen group had higher ROS and MDA(q =15.03,4.53,all P ＜ 0.05) ; the activation rate of p47phox increased significantly(x2 =19.26,P ＜ 0.05).Conclusions After oxygen exposure,p47phox translocated to membrane may regulate the NADPH oxidase-derived ROS increase in extremely premature infants.

Study on lentiviral vector target inducing p66 shc gene silencing / 重庆医学

Objective To construct p66shc gene interfering lentivirus vectors recombination and transfect it to 293T cells ,RNA interfering was carried out to induce p66shc gene silence ,so as to provide basis for further study of the p66shc function .Methods Screening of three RNA targets which were named after p66shc‐shc1 ,p66shc‐shc2 ,p66shc‐shc3 ,cloned into the pLenR‐GPH vec‐tor ,which contained green fluorescent protein(GFP) and transformed into DH5αcells .The positive clone were picked out for right sequencing and transfected to 293T cells with pRsv‐REV ,pMDlg‐pRRE ,pMD2G .The expression of GFP in inverted fluorescence microscope confirmed the virus packaging success .Fluorescence quantitative PCR and Western blot technology were used to investi‐gate the expression of p66shc at the molecular and protein levels ,p66shc‐shc1 target of effective silencing p66shc gene was selected to prepare for subsequent tests .Results The shRNA lentivirus vector was constructed which could express p66shc and was trans‐fected into 293T cells successfully .Fluorescence quantitative PCR and Western blot technology were used to investigate p66shc gene silence by RNA interference .Conclusion The lentivirus RNAi vector of targeted expression p66shc could induce p66shc gene si‐lence at the molecular and protein levels after transfected into 293T cells by RNA interference .

Silencing of Pin1 suppresses hyperoxia-induced apoptosis of A549 cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the effect of silence of Pin1 expression on hyperoxia-induced apoptosis in alveolar epithelial cells A549.</p><p><b>METHODS</b>A549 cells were divided into four groups: control, hyperoxia, negative lentivirus and Pin1-shRNA hyperoxia. The hyperoxia group was exposed to a mixture of 95%O2 and 5%CO2 for 10 minutes. Then cells were cultured in a closed environment. After 24 hours, the changes of morphology were observed under an inverted microscope. Cell apoptosis was detected by flow cytometry (FCM). The expression of X-linked inhibitor of apoptosis protein (XIAP) and Caspase-9 were detected by immunohistochemistry. The production of reactive oxygen species (ROS) and cellular mitochondria membrane potential (△Ψm) were determined by fluorescence microscopy.</p><p><b>RESULTS</b>Under the inverted microscope, the A549 cells grew slowly and the changes in morphology of the cells were most obvious in the hyperoxia and negative lentivirus groups. The changes in morphology of A549 cells were obviously improved in the Pin1-shRNA hyperoxia group. The FCM results showed that the apoptosis rate of A549 cells increased, Caspase-9 expression increased, XIAP expression decreased, mitochondrial ROS production increased and mitochondrial membrane potential decreased in the hyperoxia and negative lentivirus groups compared with the control group (P<0.05). Compared with the hyperoxia and negative lentivirus groups, the apoptosis rate of A549 cells decreased, Caspase-9 expression decreased, XIAP expression increased, mitochondrial ROS production decreased and mitochondrial membrane potential increased in the Pin1-shRNA hyperoxia group (P<0.05), although the levels of the indexes did not reach to those of the control group.</p><p><b>CONCLUSIONS</b>Silencing of Pin1 could suppress hyperoxia-induced apoptosis of A549 cells.</p>

Roles of PKCβ/P66Shc oxidative stress signal pathway in mediating hyperoxia-induced ROS production in alveolar epithelial cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the roles of PKCβ/P66Shc oxidative stress signal pathway in mediating hyperoxia-induced reactive oxgen species (ROS) production in alveolar epithelial cells (A549) and the protective effects of PKCβ inhibitor on hyperoxia-induced injuries of alveolar epithelial cells.</p><p><b>METHODS</b>A549 cells were cultured in vitro and randomly divided into three groups: control, hyperoxia and PKCβ inhibitor LY333531 treatment. The hyperoxia group was exposed to a mixture of O2 (950 mL/L) and CO2 (50 mL/L) for 10 minutes and then cultured in a closed environment. The LY333531 group was treated with PKCβ inhibitor LY333531 of 10 µmol/L for 24 hours before hyperoxia induction. Cells were collected 24 hours after culture and the levels of PKCβ, Pin1, P66Shc and P66Shc-Ser36 were detected by Western blot. The intracellular translocation of P66Shc, the production of ROS and cellular mitochondria membrane potential were measured using the confocal microscopy.</p><p><b>RESULTS</b>Compared with the control group, the levels of PKCβ, Pin1, P66Shc and P-P66Shc-Ser36 in A549 cells 24 hours after culture increased significantly in the hyperoxia group. These changes in the hyperoxia group were accompanied with an increased translocation rate of P66Shc from cytoplasm into mitochondria, an increased production of mitochondrial ROS, and a reduced mitochondrial membrane potential. Compared with the hyperoxia group, the levels of Pin1, P66Shc and P66Shc-Ser36 in A549 cells, the translocation rate of P66Shc from cytoplasm into mitochondria and the production of mitochondrial ROS decreased significantly, while the mitochondrial membrane potential increased significantly in the LY333531 treatment group. However, there were significant differences in the above mentioned measurements between the LY333531 treatment and control groups.</p><p><b>CONCLUSIONS</b>Hyperoxia can increase the expression of PKCβ in alveolar epithelial cells and production of mitochondrial ROS and decrease mitochondrial membrane potential. PKCβ inhibitor LY333531 can partially disrupt these changes and thus alleviate the hyperoxia-induced alveolar epithelial cell injury.</p>

Role of adapter protein p66Shc in injured mediating alveolar epithelial cells induced by hyperoxia / 中华实用儿科临床杂志

Objective To determine the expression of adapter protein p66Shc in mediating alveolar epithelial cells induced by hyperoxia and to explore their relationship.Methods A549 cells were cultured in vitro and divided randomly into a control group and a hyperoxia group.The hyperoxia group was exposed to a mixture of oxygen(O2,900 mL/L) and carbon dioxide(CO2,50 mL/L) for 10 min,then cultured in a closed environment.The changes in morphology were observed under inverted microscope after exposure to oxygen or air for 24 hours.The cell apoptosis was detected by flow cytometry (FC) after 24 hours.And the expression of p66Shc was detected by immunohistochemical method after 24 hours.The correlation of the changes in mitochondrial membrane potential and p66Shc protein expression was analyzed by using Bivariate correlation analysis.Results 1.Under inverted microscopy,A549 cells from the air group significantly increased,stuck to each other tightly and grew very quickly.Their adhesion was better,multy-angle oblate and many cells were in division phase.Compared with the control group,the changes in morphology of A549 were remarked and obvious than those in the hyperoxia group.The cells grew slowly,their counts decreased and the cell morphology changed from typical multi-angle oblate to round or ellipse.2.Compared with the control group,after 24 h,in hyperoxia group of A549 cells,red fluorescence decreased,and green fluorescence enhanced.3.Compared with the controls (0.057 664 88 ± 0.006 517 84),the expression of p66Shc (0.123 600 50 ± 0.004 227 23) was significantly increased in the hyperoxia group(t =-24.006,P ＜ 0.001).4.The decline of membrane potential was negatively correlated with the increased expression of p66Shc protein (R =-0.988,P ＜ 0.001).Conclusions The hyperoxia induction could significantly increase in injured mediating alveolar epithelial cells induced by hyperoxia,the expression of p66Shc increases,the membrane potential declined,and they exhibit a negative correlation.So p66Shc may be involved in the process of high oxygen damage to human alveolar epithelial cells.

The controlled study on psychosocial factors for post-stroke depression / 中国神经精神疾病杂志

Objective To investigate psychosocial factors for post-stroke depression (PSD). Methods 405 in-pa-tients with stroke were first screened for depression using Comprehensive International Diagnostic Interview-3.0. 22 pa-tients with depression were recruited as the depression group. From 383 patients without depression, 44 patients were se-lected and served as the non depression controlled group according to the sex and age paired with 1:2. Both groups were measured by using questionnaires including Life Event Scale, Simplified Coping Style Questionnaire, Social Support Rat-ing Scale, Activities of Daily Living Scale and Quality of Life Enjoyment and Satisfaction Questionnaire, Short Form. Re-sults The score of passive coping was significantly higher in depression group than in non depression group [(1.2 ± 0.5) vs. (0.8±0.7), P<0.05]. The score of subjective support was significantly lower in depression group than in non depression group [(17.5±4.0) vs. (20.7±4.6), P<0.05]. Logistic regression analysis showed, minority nationality (OR=2.564, 95%CI:1.039~6.327) and passive coping style (OR=2.223, 95%CI:1.052~5.192) were risk factors for PSD, while subjective sup-port was protective factor for PSD (OR=0.884, 95%CI:0.793~0.986). Conclusions Passive coping style and low subjec-tive support may be the important psychosocial factors of PSD.

Cloning, bioinformatic analysis, and expression of chalcone synthase gene in Carthamus tinctorius / 中草药

Objective: To clone the chalcone synthase (CHS) gene in Carthamus tinctorius, to analyze the bioinformation of CHS, to compare the expression of CHS during the florescence, and to provide the foundation for composition and regulation mechanism of the active ingredients in C. tinctorius. Methods: RNA was obtained from fresh safflower corolla, cDNA was reversely transcriped, specific primers were designed, and then CHS was cloned. The protein characteristics was analyzed using bioinformatics, the phylogenetic tree of CHS was constructed using MEGA5.1, the expression of CHS during the florescence was analyzed using real time-PCR. Results: The 1 149 bp CHS sequence in C. tinctorius was obtained, which has a 1 041 bp ORF, encoding 346 amino acids. This protein belongs to the CHS family according to Blastp in NCBI. The CHS in safflower was similar to that in above 100 plants, and the similarities to Silybum marianum, Callistephus chinensis, Chrysanthemum x morifolium, and Gynura bicolor were respectively reaching 95%, 95%, 94%, and 94%. It has the closest relationship to S. marianum according to the phylogenetic tree using MEGA5.1. The CHS formula was C1678H2693N451O493S20 and the molecular weight was 37 700, with the isoelectric point of 6.10. The number of negatively charged amino acid residues (Asp + Glu) was 42, and the number of positively charged amino acid residues (Arg + Lys) was 38. The gene expression analysis showed that the highest expression of CHS in safflower was on day 3 of florescence, much higher than that on the other days. Conclusion: The CHS in safflower is successfully cloned, analyzed, and expressed, which provides the foundation for composite and regulation mechanism of the active ingredients in C. tinctorius.

Efficacy and safety of short-term intensive statin therapy in patients with acute coronary syndrome / 中华全科医师杂志

Objective To evaluate the clinical efficacy and safety of short-term intensive statin therapy in patients with acute coronary syndrome ( ACS) .Methods A total of 218 ACS patients admitted in Hangzhou First People′s Hospital from March 2013 to July 2013 were enrolled into this study .The patients were randomly assigned to receive atorvastatin 80 mg/d during hospitalization , and 40 mg/night after discharge for one month ( intensive group , n=107 );or receive atorvastin 20 mg during hospitalization and 20 mg/night after discharge for one month ( control group, n =111 ).The biochemical indexes were measured on the admission and after one-month treatment.Results After one-month treatment, the total cholesterol, triglycerides and LDL cholesterol of intensive group were significantly lower , and the high density lipoprotein cholesterol was higher than baseline values ( 0.75 ±0.14 ) mmol/L vs.( 1.52 ±0.88 ) mmol/L, P<0.05;(2.21 ±0.78)mmol/L vs.(4.55 ±1.12)mmol/L, P<0.05;(1.76 ±0.31)mmol/L vs.(2.23 ±0.77) mmol/L, P<0.05; (1.15 ±0.34) vs.(1.52 ±0.41) mmol/L, P<0.05.The liver enzymes creatine kinase in intensive group was not significantly changed , but the creatinine levels decreased (82.53 ±23.85)μmol/L vs.(57.81 ±15.27) μmol/L, P<0.05, and the blood homocysteine and ultra-sensitivity C-reactive protein levels also decreased compared with the baseline ( 10.52 ±4.66 ) mmol/L vs.(30.70 ±18.82 ) mmol/L, P <0.05;( 8.06 ±2.68 ) mg/L vs.( 19.75 ±11.91 ) mg/L, P <0.05. Conclusions Short-term intensive statin therapy can effectively reduce blood lipid , cholesterol and homocysteine levels and raise HDL cholesterol levels; also with its anti-inflammatory and renal protective effect the therapy can provide more clinical benefit for patients with ACS .