Objective:To investigate the effect of DEAD-box helicase (DDX) 21 on myocardial ischemia-reperfusion (I/R) injury and its potential mechanisms.Methods:In vivo, adult male Bama pigs and C57BL/6J mice were used to establish a myocardial I/R injury model by ligating the left anterior descending coronary artery, with sham-operated groups set as controls. The expression of DDX21 in myocardium after I/R injury was assessed by quantitative real-time PCR (qRT-PCR), Western blot, and immunofluorescence staining. Following the establishment of the myocardial I/R injury model in mice, AAV9 vectors with cardiac-specific expression were injected in situ into the peri-infarct region (The I/R+DDX21 group, I/R+negative control (NC) group, I/R+sh-NC group and I/R+sh-DDX21 group were injected with AAV9:cTnT-DDX21, AAV9:cTnT-NC, AAV9:cTnT-sh-NC and AAV9:cTnT-sh-DDX21, respectively). Additionally, the I/R+A-485 group received intraperitoneal injections of the cAMP response element-binding protein (CREB) binding protein inhibitor A-485, while the I/R+PBS group was injected with an equivalent volume of phosphate-buffered saline (PBS) as the control. Echocardiography was performed on postoperative days 1 and 28 to evaluate cardiac function (left ventricular ejection fraction and fractional shortening). At 28 days post-surgery, mice were euthanized and heart tissues were harvested for histological sectioning. Myocardial fibrosis was evaluated using Masson′s trichrome staining. In vitro, primary cardiomyocytes were isolated from neonatal day 1 C57BL/6J mice using enzymatic digestion method. Cardiomyocytes were transfected with plasmids or small interfering RNA (siRNA). The cardiomyocytes transfected with DDX21-siRNA were assigned to the siDDX21 group, those transfected with the DDX21 plasmid were assigned to the DDX21 group, and those transfected with the corresponding empty plasmid or siRNA were assigned to the NC group. Additionally, cardiomyocytes were treated with A-485 (A-485 group) or PBS (PBS group). An oxygen-glucose deprivation/reoxygenation (OGD/R) model was used to simulate cellular injury. Transcriptome sequencing was performed to identify downstream mechanisms of DDX21. Differential gene expression analysis was conducted using software such as DESeq2, and alternative splicing events in the mRNA transcriptome were analyzed using rMATS software. Mitochondrial superoxide, mitochondrial membrane potential, ATP content, and mitochondrial respiratory chain complex enzyme activity in cardiomyocytes were detected using immunofluorescence staining and commercial assay kits. The oxidative phosphorylation level of the cells was assessed by the Seahorse extracellular flux analyzer. Acetylated DDX21 levels were measured using co-immunoprecipitation and Western blot assays.Results:The expression levels of DDX21 in myocardium from the Bama pigs and mice in the I/R injury model were significantly higher than those in the sham group (all P<0.001). Echocardiographic results showed that at 28 days post-surgery, compared to the I/R+NC group, the I/R+DDX21 group exhibited higher left ventricular ejection fraction and fractional shortening, while the I/R+sh-DDX21 group showed lower values; Masson staining results demonstrated that, compared to the I/R+NC group, the myocardial fibrosis area in the I/R+DDX21 group was significantly reduced, whereas it was significantly increased in the I/R+sh-DDX21 group (all P<0.001). Transcriptomic sequencing results suggested that DDX21 may influence myocardial injury by regulating mitochondrial metabolic activity. In vitro, compared to the OGD/R+NC group, the OGD/R+DDX21 group exhibited lower mitochondrial superoxide levels, higher polymer/monomer ratio, maximal oxygen consumption, reserve capacity, and ATP content. In contrast, the OGD/R+siDDX21 group showed the opposite results, with reduced activity of mitochondrial respiratory chain complex V (all P<0.05). Mechanistically, rMATS software and other analyses indicated that knockdown of DDX21 affected the alternative 3′ splicing sites of ATP5J precursor mRNA, inhibiting the splicing of certain exonic sequences. Overexpression of DDX21 upregulated both mRNA and protein levels of ATP5J. Co-immunoprecipitation experiments showed that, compared to the PBS group, acetylated DDX21 levels were reduced in the A-485 group. Further in vivo experiments showed that, compared to the I/R+PBS group, the I/R+A-485 group exhibited higher left ventricular ejection fraction and fractional shortening, and a lower proportion of left ventricular fibrosis (all P<0.001). Conclusions:DDX21 improves cardiomyocyte energy metabolism and alleviates I/R injury by regulating the alternative splicing of ATP5J. A-485 holds potential as a novel small molecule candidate for the treatment of myocardial injury.

Objective:To investigate the effect of DEAD-box helicase (DDX) 21 on myocardial ischemia-reperfusion (I/R) injury and its potential mechanisms.Methods:In vivo, adult male Bama pigs and C57BL/6J mice were used to establish a myocardial I/R injury model by ligating the left anterior descending coronary artery, with sham-operated groups set as controls. The expression of DDX21 in myocardium after I/R injury was assessed by quantitative real-time PCR (qRT-PCR), Western blot, and immunofluorescence staining. Following the establishment of the myocardial I/R injury model in mice, AAV9 vectors with cardiac-specific expression were injected in situ into the peri-infarct region (The I/R+DDX21 group, I/R+negative control (NC) group, I/R+sh-NC group and I/R+sh-DDX21 group were injected with AAV9:cTnT-DDX21, AAV9:cTnT-NC, AAV9:cTnT-sh-NC and AAV9:cTnT-sh-DDX21, respectively). Additionally, the I/R+A-485 group received intraperitoneal injections of the cAMP response element-binding protein (CREB) binding protein inhibitor A-485, while the I/R+PBS group was injected with an equivalent volume of phosphate-buffered saline (PBS) as the control. Echocardiography was performed on postoperative days 1 and 28 to evaluate cardiac function (left ventricular ejection fraction and fractional shortening). At 28 days post-surgery, mice were euthanized and heart tissues were harvested for histological sectioning. Myocardial fibrosis was evaluated using Masson′s trichrome staining. In vitro, primary cardiomyocytes were isolated from neonatal day 1 C57BL/6J mice using enzymatic digestion method. Cardiomyocytes were transfected with plasmids or small interfering RNA (siRNA). The cardiomyocytes transfected with DDX21-siRNA were assigned to the siDDX21 group, those transfected with the DDX21 plasmid were assigned to the DDX21 group, and those transfected with the corresponding empty plasmid or siRNA were assigned to the NC group. Additionally, cardiomyocytes were treated with A-485 (A-485 group) or PBS (PBS group). An oxygen-glucose deprivation/reoxygenation (OGD/R) model was used to simulate cellular injury. Transcriptome sequencing was performed to identify downstream mechanisms of DDX21. Differential gene expression analysis was conducted using software such as DESeq2, and alternative splicing events in the mRNA transcriptome were analyzed using rMATS software. Mitochondrial superoxide, mitochondrial membrane potential, ATP content, and mitochondrial respiratory chain complex enzyme activity in cardiomyocytes were detected using immunofluorescence staining and commercial assay kits. The oxidative phosphorylation level of the cells was assessed by the Seahorse extracellular flux analyzer. Acetylated DDX21 levels were measured using co-immunoprecipitation and Western blot assays.Results:The expression levels of DDX21 in myocardium from the Bama pigs and mice in the I/R injury model were significantly higher than those in the sham group (all P<0.001). Echocardiographic results showed that at 28 days post-surgery, compared to the I/R+NC group, the I/R+DDX21 group exhibited higher left ventricular ejection fraction and fractional shortening, while the I/R+sh-DDX21 group showed lower values; Masson staining results demonstrated that, compared to the I/R+NC group, the myocardial fibrosis area in the I/R+DDX21 group was significantly reduced, whereas it was significantly increased in the I/R+sh-DDX21 group (all P<0.001). Transcriptomic sequencing results suggested that DDX21 may influence myocardial injury by regulating mitochondrial metabolic activity. In vitro, compared to the OGD/R+NC group, the OGD/R+DDX21 group exhibited lower mitochondrial superoxide levels, higher polymer/monomer ratio, maximal oxygen consumption, reserve capacity, and ATP content. In contrast, the OGD/R+siDDX21 group showed the opposite results, with reduced activity of mitochondrial respiratory chain complex V (all P<0.05). Mechanistically, rMATS software and other analyses indicated that knockdown of DDX21 affected the alternative 3′ splicing sites of ATP5J precursor mRNA, inhibiting the splicing of certain exonic sequences. Overexpression of DDX21 upregulated both mRNA and protein levels of ATP5J. Co-immunoprecipitation experiments showed that, compared to the PBS group, acetylated DDX21 levels were reduced in the A-485 group. Further in vivo experiments showed that, compared to the I/R+PBS group, the I/R+A-485 group exhibited higher left ventricular ejection fraction and fractional shortening, and a lower proportion of left ventricular fibrosis (all P<0.001). Conclusions:DDX21 improves cardiomyocyte energy metabolism and alleviates I/R injury by regulating the alternative splicing of ATP5J. A-485 holds potential as a novel small molecule candidate for the treatment of myocardial injury.

Objective To investigate the effect of pronunciation organ correction combined with language training on rehabilitation in children with cerebral palsy and language disorder.Methods A total of 120 cases of pe-diatric cerebral palsy and language disorder treated with rehabilitation in our hospital from January 2015 to March 2018 were divided into routine group and study group by a random digital table,with 60 cases in each group.The routine group was given language training alone,while the study group was given pronunciation organ correction combined language training.The assessment results of children with dysarthria and language retardation before and after treatment were compared respectively,and the efficacy and family satisfaction were compared between the two groups.Results There were statistical differences in the assessment results of children with dysarthria and lan-guage retardation in both groups after treatment compared with before treatment(P＜0.05),and there were statis-tical differences between the two groups after treatment(P＜0.05).The overall clinical efficiency was 75％in the study group,which was higher than that of 60％in the routine group(P＜0.05).The total satisfaction rate was 90％in the study group,which was higher than that of 75％in the routine group(P＜0.05).Conclusion Giving pronunciation organ correction combined language training in the rehabilitation treatment of pediatric cerebral palsy and language disorder can improve treatment efficacy and the satisfaction of child family members.

Objective To evaluate the effects of dexmedetomidine on NLRP3 inflammasome during acute lung injury ( ALI ) induced by blunt chest trauma and hemorrhagic shock-resuscitation in rats. Methods Forty-five SPF healthy male Sprague-Dawley rats, weighing 200-220 g, were divided into 3 groups (n=15 each) using a random number table method: sham operation group (Sham group), ALI group and dexmedetomidine group ( Dex group) . A rat model of ALI was established by blunt chest trauma and hemorrhagic shock-resuscitation. Dexmedetomidine 5 μg·kg-1 ·h-1 was infused into the femoral vein after blunt chest trauma in Dex group. Blood samples were collected from the femoral artery at 6 h after blunt chest trauma for measurement of arterial oxygen partial pressure and concentrations of pro-inflammatory cytokines interleukin-1beta ( IL-1β) and IL-18 in serum ( by enzyme-linked immunosorbent assay ) . The oxygenation index was calculated. The rats were sacrificed and lungs were removed for examination of the pathological changes which were scored and for determination of the expression of NLRP3, caspase-1 and apoptosis-associated speck-like protein containing CARD ( ASC) in lung tissues ( by Western blot) and ac-tivities of lactic dehydrogenase ( LDH) and myeloperoxidase ( MPO) in lung tissues ( by colorimetry) . Re-sults Compared with Sham group, PaO2 and oxygenation index were significantly decreased, the expres-sion of NLRP3, caspase-1 and ASC was up-regulated, the activities of LDH and MPO were increased, and the concentrations of IL-1β and IL-18 in serum were increased in ALI group ( P＜0. 05 ) . Compared with ALI group, PaO2 and oxygenation index were significantly decreased, the expression of NLRP3, caspase-1 and ASC was down-regulated, the activities of LDH and MPO were decreased, and the concentrations of IL-1β and IL-18 in serum were decreased in ALI group ( P＜0. 05) . Conclusion Dexmedetomidine can al-leviate blunt chest trauma and hemorrhagic shock-resuscitation induced ALI in rats, and the mechanism is related to inhibition of NLRP3 inflammasome activation and reduction of inflammatory response.

Objective: To analyze 167 batches of Weiling granules according to the quality standard and exploratory research, and evaluate the overall quality and standard condition of the preparation. Methods: A TLC method was established for the identification of Atractylodis macrocephalae Rhizoma;an HPLC method was established for the fingerprint and the content determination of paeoniflorin, tetrahydropalmatine and ammonium glycyrrhizinate. An Agilent Poroshell120,SB-C18analytical column (100 mm×4. 6 mm, 2. 7 μm) was employed with gradient elution of acetonitrile-0. 1% phosphoric acid as the mobile phase at the flow rate of 1. 8 ml·min-1, and the sample size was 3 μl. Acid base titration method was used for measuring acid-neutralizing capacity. Results: No interference from the negative controls was shown to the TLC identification of Atractylodis macrocephalae Rhizoma. The fingerprint exhibited better separation of each peak. The precision, reproducibility and stability of the method were good,and the RSDs of the relative retention time and rela-tive peak area were less than 3. 0% . The linear range of paeoniflorin, tetrahydropalmatine and ammonium glycyrrhizinate was 0. 057-0. 568 μg(r=0. 999 9), 0. 035-0. 353 μg(r=0. 999 9)and 4. 244×10 -3-42. 44×10 -3μg(r=0. 999 9), respectively, and the av-eragerecoverywas99.3%(RSD=1.0%,n=6),100.0%(RSD=0.8%,n=6) and99.8%(RSD=1.2%,n=6),respectively. The average recovery of acid-neutralizing capacity was 99. 5% (RSD=0. 5% ,n=6). Conclusion: Exploratory research increases the specificity, controllability and safety of the standards, which provides reference for the further drug standards revision and the drug quality control.

Objective To establish a pre-audit system for medical orders in hospitalized patients and increase the auditing rate in inpatients. Methods In the pre-audit system for medical orders, browser/server(B/S)and client/server(C/S)hybrid architecture was adopted and JAVA language was used for programming. The pre-audit system for medical orders was linked to the hospital information system and data integration platform,and audit module was improved on the basis of the pre-audit system for prescriptions which had been successfully operated in Xuanwu Hospital of Capital Medical University. More than 20 thousand related rules were added in the audit think tank for rational drug use of prescriptions and serious errors interception for indications,dosage and administration route,appropriate translation of Chinese patent medicine indications,refinement of compatibility taboos,audit of drug allergy risk,appropriate audit for pain medications,and audit for rational drug use related to height,body weight,body surface area,and test values were performed. The audit think tank for rational drug use was formed,which set up the foundation for intelligent examination of medical orders. The medical orders which were not passed by intelligent examination were transmitted to the pre-audit pharmacist to audit for the second time. By comparing the audit rate of medical orders in hospitalized patients before and after adopting the pre-audit system and the qualification rate at the first 4 months after adopting the system,the application effect was preliminarily evaluated. Results After launching pre-audit system for medical orders in June 2017,the examination rate of medical orders in inpatients increased from 42.53%(61 139/143 756)to 100%(at the first 4 months,the number of doctor′s orders was 69 183,74 072,99 816,and 184 184,respectively). At the first 4 months after the system was launched,the pass rate of audit for medication orders was 69.02%(47 753/69 183),84.27%(62 416/74 072),92.69%(92 516/99 816),and 97.21%(179 040/184 184),respectively. The difference in passing rate of audit for medication orders between the 4th month and the first month was statistically significant(χ2=42 548.86,P<0.001). Conclusions The pre-audit system for inpatients′medication orders was successfully established. The intelligent audit for all inpatients realized using this pre-audit system and was of benefit to the rational drug use improvement in hospitals.

Objective To establish a pre-audit system for medical orders in hospitalized patients and increase the auditing rate in inpatients. Methods In the pre-audit system for medical orders, browser/server(B/S)and client/server(C/S)hybrid architecture was adopted and JAVA language was used for programming. The pre-audit system for medical orders was linked to the hospital information system and data integration platform,and audit module was improved on the basis of the pre-audit system for prescriptions which had been successfully operated in Xuanwu Hospital of Capital Medical University. More than 20 thousand related rules were added in the audit think tank for rational drug use of prescriptions and serious errors interception for indications,dosage and administration route,appropriate translation of Chinese patent medicine indications,refinement of compatibility taboos,audit of drug allergy risk,appropriate audit for pain medications,and audit for rational drug use related to height,body weight,body surface area,and test values were performed. The audit think tank for rational drug use was formed,which set up the foundation for intelligent examination of medical orders. The medical orders which were not passed by intelligent examination were transmitted to the pre-audit pharmacist to audit for the second time. By comparing the audit rate of medical orders in hospitalized patients before and after adopting the pre-audit system and the qualification rate at the first 4 months after adopting the system,the application effect was preliminarily evaluated. Results After launching pre-audit system for medical orders in June 2017,the examination rate of medical orders in inpatients increased from 42.53%(61 139/143 756)to 100%(at the first 4 months,the number of doctor′s orders was 69 183,74 072,99 816,and 184 184,respectively). At the first 4 months after the system was launched,the pass rate of audit for medication orders was 69.02%(47 753/69 183),84.27%(62 416/74 072),92.69%(92 516/99 816),and 97.21%(179 040/184 184),respectively. The difference in passing rate of audit for medication orders between the 4th month and the first month was statistically significant(χ2=42 548.86,P<0.001). Conclusions The pre-audit system for inpatients′medication orders was successfully established. The intelligent audit for all inpatients realized using this pre-audit system and was of benefit to the rational drug use improvement in hospitals.

Objective To evaluate the effects of apolipoprotein-J(ApoJ)on hypoxia/reoxygenation(H/R)induced neonatal rat ventricular cells(NRVCs)injury and its related signaling pathways.Methods ApoJ high expression was achieved by infection with recombinant adenovirus in NRVCs.The three gas incubator was used to establish H/R model,SOD mimetic Mn(Ⅲ) tetrakis(4-benzoic acid)porphyrin chloride and eIF2α dephosphory inhibitor Salubrinal were performed the pretreatment.The NRVCs were divided into four groups:normal control group,H/R,ApoJ group,ApoJ+ H/R group,Mn(Ⅲ)TBAP+H/R group and Salubrinal +-H/R group.The cell viability was measured by MTT assay;the leakages of LDH,the expression of SOD and caspase-3/7 activity were detected by ELISA.The protein expression levels of ApoJ,Nox2/gp91phox,caspase-12,CHOP,and the phosphorylation level of eukaryotic initiation factor 2α(eIF2α)were determined by Western blot.Results ApoJ protein in myocardial cells was highly expressed after infection by recombinant adenoviru.Compared with the control group,the cell viability and the activity of SOD were significantly decreased,the leakages of LDH and the activity of caspase-3/7 were increased in the H/R group,the pro tein expression level of Nox2/gp91phox,caspase-12 and CHOP and the phosphorylation level of eIF2α were increased.Compared with the H/R group,the leakages of LDH and the activity of caspase-3/7 in the ApoJ overexpression group,Mn(Ⅲ)TBAP group and Salubirnal group were significantly decreased.ApoJ overexpression significantly increased the cell viability and the activity of SOD.Moreover,the protein expression level of Nox2/gp91phox,caspase-12 and CHOP were significantly decreased,while the phosphorylation level of eIF2α was markedly increased.Conclusion ApoJ alleviates H/R induced myocardial cellular injury by antioxidative stress and endoplasmic reticulum stress.

Interferon beta (IFN-beta) and TNF-related apoptosis-inducing ligand (TRAIL) are effective anticancer agents. Adeno-associated virus (AAV) is one of the current most promising gene delivery vectors. Previously, we constructed tumor-targeting AAV-hTERT-IFN-beta and AAV-hTERT-TRAIL by inserting IFN-beta or TRAIL gene into AAV controlled by hTERT promoter. The studies showed that either single IFN-beta or TRAIL gene therapy exhibited a certain extent anticancer effect. Here, we report their inhibitory effects on A549 lung cancer cell growth in vitro and in vivo by combined AAV-hTERT-IFN-beta and AAV-hTERT-TRAIL. Expression of secreted IFN-beta in lung cancer A549 cells infected by AAV-hTERT-IFN-beta was detected by enzyme-linked immunosorbent assay (ELISA). The growth-suppressing effect of AAV-hTERT-IFN-beta in combination with AAV-hTERT-TRAIL on several cancer cell lines was assessed by MTT assay. Apoptosis of A549 cancer cells infected by AAV-hTERT-IFN-beta alone, AAV-hTERT-TRAIL alone, and their combination was evaluated by apoptotic cell staining and flow cytometry (FCM), respectively. The antitumor effect of the combination of AAV-hTERT-IFN-beta with AAV-hTERT-TRAIL in vivo was further evaluated through A549 lung cancer xenograft in nude mice. The results showed that the combinational treatment was superior to any alone and presented intensified tumor cytotoxic and apoptotic effect on A549 cancer cells. Most importantly, the combination of AAV-hTERT-IFN-beta with AAV-hTERT-TRAIL exhibited significant antitumor effect and eliminated all tumor masses in nude mice, which lay a foundation for exploring the molecular mechanisms of combined IFN-beta and TRAIL anti-tumor activity.
Animals ; Antineoplastic Agents ; metabolism ; pharmacology ; Cell Line, Tumor ; Dependovirus ; genetics ; metabolism ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Interferon-beta ; genetics ; pharmacology ; Lung Neoplasms ; pathology ; therapy ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; pharmacology

Several digital filtering techniques for correcting baseline drift from ECG signals are presented. A bidirectional filter is introduced, which can cantain baseline drift their filtering performance and realization methods are compared.
Electrocardiography ; instrumentation ; methods ; Filtration ; instrumentation ; Signal Processing, Computer-Assisted ; instrumentation