The study aims at cloning the CDS fragment of erk2 gene cDNA in Inner Mongolia Cashmere Goat and analyzing its tissue-specific expression, erk2 gene cDNA was cloned by RT-PCR. The nucleotide sequence was analyzed by Blast and amino acid sequence was analyzed by online softwares SMART and Psite. The tissue-specific expression pattern of erk2 was analyzed by quantitative RT-PCR. The expression of erk2 in testis of goat was detected by Immunohistochemistry. The cloned erk2 gene cDNA (GenBank Accession No. JX569765) was 1 083 bp in length, including a complete ORF encoding 360 amino acids residues. The amino acid sequence shares 100% identity with the Bos Taurus ERK2 (Bos Taurus BC133588.1). Analysis by SMART suggests that the encoded protein contained a "TEY" structure and an S-TKc domain possessing serine/threonine kinase catalytic activity. Analysis with Psite indicates one cAMP-/cGMP-dependent protein kinase phosphorylation site, 3 protein kinase C phosphorylation sites, 5 casein kinase II phosphorylation sites, 2 protein kinases ATP-binding region signatures and one serine/threonine protein kinases active-site signature in this protein. Analysis by Psort (k-NN prediction) suggestes that this protein most probably is localized in cytoplasm. The results of quantitative RT-PCR show that the expression of erk2 mRNA was higher in heart, skin and breast, whereas lower in spleen and kidney. ERK2 protein was detected in testis by immunohistochemistry.
Amino Acid Sequence ; Animals ; China ; Cloning, Molecular ; DNA, Complementary ; genetics ; Goats ; genetics ; Male ; Mitogen-Activated Protein Kinase 1 ; genetics ; metabolism ; Molecular Sequence Data ; Real-Time Polymerase Chain Reaction ; Testis ; metabolism

Objective To evaluate the impact of sarpogrelate on the in-stent restenosis (ISR) after percutaneous transluminal angioplasty (PTA) in peripheral arterial diseases (PAD).Methods PAD patients who had PTA for the first time were divided into two groups receiving respectively clopidogrel and aspirin (clopidogrel group) or sarpogrelate and aspirin (sarpogrelate group).Vascular ultrasonography was performed at 6 months after PTA in all patients to evaluate the degree of ISR.Results 62 patients finished the follow-up visits as required.The in-stent restenosis in the sarpogrelate group and clopidogrel group was 7.0％ vs.18.1％ (P =0.036),the peak systolic velocity ratio was 1.34 vs.2.08 (P =0.010) and the cases of ISR was 1 vs.10 (P =0.005) respectively.No patients reported serious adverse events.Conclusions Sarpogrelate combined with aspirin is safe and effective.Compare with clopidogrel and aspirin,sarpogrelate and aspirin can significantly reduce the rate of ISR after PTA and the intimal proliferation in the stent.

Objective:To investigate the difference of accuracy between magnetic induction Freehand-3D ultrasound and two-dimensional ultrasound in measuring the volume of thyroid model.Methods:Forty thyroid models were established using porcine liver, and the Archimedes procedure was set as gold standard in the measurement of the volume of each model. The accuracy of measurement of the porcine thyroid model volume between two-dimensional ultrasound and magnetic induction Freehand-3D ultrasound were compared.Results:There were no significant differences in the accuracy of measurements of thyroid model volume among two-dimensional ultrasound, magnetic induction Freehand-3D ultrasound and Archimedes procedure (all P>0.05). Compared with the Archimedes procedure, magnetic induction Freehand-3D ultrasonic method showed higher correlation coefficient of the measurement of thyroid model volume ( r=0.998). Bland-Altman analysis showed the lower measure error with a relative error of 3.42% and range of -9.57% to 12.07%. And the limits of agreement were (-1.253, 0.999) in the magnetic induction Freehand-3D ultrasonic measurement. Conclusions:Compared with two-dimensional ultrasound, the magnetic induction Freehand-3D ultrasound show higher accuracy in the measurement of the volume of the thyroid model.

AIM:To study the effects of noninvasive delayed limb ischemia preconditioning ( NDLIP) on ani-mal cardiac function , myocardial morphology and myocardial apoptosis after myocardial infarction ( MI ) .METHODS:Healthy SD male rats [n=45, weighing (250 ±10) g] were randomly divided into 3 groups:MI group:the animal model of MI was established by surgical ligation of left anterior descending artery ( LAD) after 2 weeks;NDLIP group:after the success of the MI animal model , NDLIP was carried out every other day until the 4th, 6th and 8th weeks;sham group:as the negative control group , the animals were taken heart LAD threading but no ligation .All rats were fed conventionally .At the end of the 4th, 6th and 8th weeks, all rats were made ventricular intubation , and then the hemodynamic parameters were recorded .The blood samples were withdrawn from the abdominal aorta and the serum was separated via centrifugation . The serum contents of Bcl-2 and Bax were measured by ELISA .Left ventricular anterior wall was homogenized .The mito-chondrial respiratory chain complexes Ⅰ,Ⅱ,Ⅲand Ⅳin the myocardial tissues were detected by ELISA .RESULTS:At the end of the 4th, 6th and 8th weeks, compared with MI group, left ventricular systolic pressure in NDLIP group was significantly increased , while left ventricular end-diastolic pressure in NDLIP group was significantly decreased ( both P<0.05).Mitochondrial respiratory chain complexesⅠ, Ⅱ, Ⅲ and Ⅳ in NDLIP group were significantly increased (P<0.05).The serum level of Bcl-2 in NDLIP group was significantly increased and Bax level was reduced remarkably (both P<0.01) .CONCLUSION:NDLIP improves the hemodynamic indexes , promotes the mitochondrial respiratory function and inhibits cell apoptosis , thus improving the prognosis of MI .

Aim To study the preventative effects of noninvasive delayed limb ischemic preconditioning ( NDLIP) on sudden cardiac death in rats with myocar-dial infarction. Methods Thirty healthy SD male rats weighting ( 250 ± 10 ) g were randomly divided into 3 groups:① myocardial infarction ( MI ) group: animal model of MI was established by making surgical ligation of animal LAD. ② MI plus NDLIP group: after the success of the animal model of MI, NDLIP was carried out every other day until 4th week. ③Sham group:as the negative control group, animals were taken heart LAD threading but no ligation. All rats were fed con-ventionally. At the end of 4 weeks, three groups of rats were administered with metaraminol ( 0. 2 mg · min-1 ) . ECG, drug cumulant of sudden death and death onset time were recorded. After sudden death, blood samples were withdrawn from abdominal aorta and serum was separated via centrifugation. ELISA method was used to measure serum caspase-3 , HSP70 and SOD concentration. Results While metaraminol led animal cardiac sudden death, the rats heart rate ( HR) kept declining with the increase of dosage of metaraminol during the administration period. Rat HR of MI+NDLIP group [ ( 479 ± 8 ) vs ( 416 ± 19 ) beat ·min-1 , ( 446 ± 32 ) vs ( 370 ± 20 ) beat · min-1 , (376 ± 53) vs (305 ± 29) beat·min-1, (307 ± 63) vs (244 ± 33) beat·min-1, (283 ± 45) vs (121 ± 35 ) beat · min-1 , P <0. 01 ] was markedly higher than that of MI group at 0 , 5 , 10 , 30 , 50 min before death. Compared with MI group, drugs cumulant to sudden death and death onset time of MI + NDLIP group [ ( 14. 58 ± 3. 03 ) vs ( 10. 76 ± 2. 73 ) mg, (72. 9 ± 15. 2 ) vs ( 53. 8 ± 13. 6 ) min, P <0. 01 ] were significantly increased. Compared with MI group, serum caspase-3 content of MI+NDLIP group was sig-nificantly reduced [ ( 2. 01 ± 0. 52 ) vs ( 2. 34 ± 0. 38 )μg·L-1 , P<0. 01 ]; HSP70 levels were remarkably increased [ ( 3. 01 ± 0. 58 ) vs ( 2. 70 ± 0. 43 ) μg · L-1 , P <0. 05 ]; SOD levels were significantly im-proved [(1. 99 ± 0. 65) vs (1. 70 ± 0. 58) mg·L-1, P<0. 01 ] . Conclusion NDLIP can prevent sudden cardiac death after myocardial infarction in rats, which may be mediated by reducing the myocardial cell apop-tosis, increasing protective protein expression and en-hancing antioxidant capacity.

Objective: To investigate the current status of occupational mental health and job satisfaction in university teachers in Shenyang, China and related influencing factors. Methods: A total of 1500 teachers from 6 universities in Shenyang were randomly selected as study subjects from November 2013 to January 2014. Self-administered questionnaires were used to investigate mental health, including effort-reward imbalance questionnaire, Minnesota Satisfaction Questionnaire, Center for Epidemiological Survey-Depression Scale, and Psychological Capital Questionnaire. Results: Of all teachers, 58.9% had depressive symptoms. Depressive symptoms in university teachers were negatively correlated with the scores of psychological capital, supervisory commitment, and perceived organizational support (r=-0.461, -0.306, and -0.366, all P<0.01) and were positively correlated with the score of occupational stress (r=0.414, P<0.01) . Job satisfaction was positively correlated with psychological capital, perceived organizational support, and supervisory commitment (r=0.650, 0.715, and 0.636, all P<0.01) and negatively correlated with occupational stress (r=-0.475, P<0.01) . The direct effects of occupational stress, perceived organizational support, and supervisory commitment on job satisfaction were -0.30, 0.26, and 0.14, respectively, and their indirect effects were -0.0176, 0.0656, and 0.0368, respectively. The direct effects of occupational stress, perceived organizational support, and supervisory commitment on depressive symptoms were 0.20, -0.08, and -0.05, respectively, and their indirect effects was 0.033, -0.123, and -0.069, respectively. Conclusion Occupational mental health is closely associated with job satisfaction in university teachers in Shenyang, and psychological capital has a mediating effect on perceived organizational support, supervisory commitment, occupational stress, job satisfaction, and depressive symptoms.

Objective:To investigate the effect of stress-induced protein Sestrin2 (SESN2) on necroptosis of mouse dendritic cell (DC) induced by lipopolysaccharide (LPS) combined with zVAD, a panaspartate-specific cysteine protease (caspase) inhibitor.Methods:The DC2.4 cell line derived from the bone marrow of mouse in the 3rd to 10th generations was cultured. The cells were stimulated with LPS for 0 hour, 6 hours, 12 hours, and 24 hours, and grouped according to the stimulation time points. Western blotting was performed to determine the protein expression of SESN2 in each group. Overexpression empty lentivirus (NC), SESN2 gene overexpression RNA sequence lentivirus (SESN2 LV-RNA), small interfering empty lentivirus (NS), and SESN2 gene small interfering RNA sequence lentivirus (SESN2 siRNA) were transfected into DC2.4 cells. After 72 hours of transfection, cell fluorescence expression was observed under the inverted fluorescence microscope. Cells in each transfection group were stimulated with LPS for 24 hours. The blank control groups were set up and cultured with phosphate buffered saline (PBS) for 24 hours. Western blotting was performed to measure SESN2 protein expression. In the same groups as above, cells were stimulated with LPS+zVAD for 24 hours. The blank control groups were set up and cultured with PBS for 24 hours. Western blotting was used to determine the expression of mixed lineage kinase domain-like protein (MLKL) and phosphorylated-MLKL (p-MLKL). The p-MLKL levels and the number of positive cells were observed using laser scanning confocal microscopy. The necroptotic cell ratios were assessed by both flow cytometry and Hoechst staining.Results:Compared to the LPS 0 hour group, the expression of SESN2 in the LPS 24 hours group showed a significant increase. Therefore, 24 hours was chosen as the subsequent stimulation time point. After successful lentivirus transduction and 24 hours of cultivation, the MLKL phosphorylation level in the SESN2 siRNA+LPS+zVAD group was significantly higher than that in the NS+LPS+zVAD group. The MLKL phosphorylation in the SESN2 LV-RNA+LPS+zVAD group was significantly lower than that in the NC+LPS+zVAD group. The MLKL phosphorylation levels in both the NS+LPS+zVAD group and the NC+LPS+zVAD group were obviously higher than those in the NS+PBS group and the NC+PBS group, respectively. Laser scanning confocal microscopy showed that the trends in quantity and fluorescence intensity of p-MLKL protein expressions were consistent with the above results. The results from flow cytometry analysis and Hoechst staining showed that the rates of cell necrotic apoptosis in SESN2 siRNA+LPS+zVAD group were significantly higher than those in NS+LPS+zVAD group [flow cytometry analysis: (30.800±1.153)% vs. (20.800±1.114)%, Hoechst staining: (75.267±0.451)% vs. (46.267±3.371)%, both P < 0.05], indicating that knocking down SESN2 further exacerbated the occurrence of necroptosis. The necrotic apoptosis rates in SESN2 LV-RNA+LPS+zVAD group were significantly lower than those in NC+LPS+zVAD group [flow cytometry analysis: (7.160±0.669)% vs. (19.240±2.322)%, Hoechst staining: (32.433±3.113)% vs. (48.567±4.128)%, both P < 0.05], indicating that overexpressing SESN2 reversed such response and markedly reduced the proportion of necroptotic cells compared to the corresponding empty vector group. Conclusion:SESN2 exhibits an inhibitory effect on necroptosis of DC in sepsis. Targeted SESN2 expression may regulate the process of DC-mediated immune response in sepsis.

Malignant tumors of the digestive system , with high morbidity and mortality rate , are insidious at the onset and lack of effective treatments so far .Interleukin-22 (IL-22) is one of the members of the IL-10 cytokine family discovered in recent years and was originally called IL-10-associated T cell differentiation inducing factor (IL-TIF).IL-22 expression is elevated in various digestive system malignant tumors, and increased IL-22 expression is associated with tumor progression and poor prognosis .Studies on the molecular mechanism revealed that IL-22 initiates a series of downstream signaling pathways such as JAK/STAT and MAPK, by acting on the IL-22 receptor, inducing tumorigenesis.IL-22 is expected to be a novel diagnostic biomarker and therapeutic target of digestive system malignant tumor .

OBJECTIVE: To determine the frequency of chromosomal abnormalities and outcome of pregnancy for fetuses with increased nuchal translucency (NT). METHODS: Between July 2014 and February 2018, 247 fetuses with increased NT (>95th centile)were analyzed by chromosome microarray analysis (CMA). The fetuses were divided into ones with isolated increased NT (168 cases), increased NT with cystic hygroma (20 cases), increased NT with edema (12 cases) or increased NT with other abnormalities (47 cases). All couples were followed up by telephone calls. RESULTS: The rate of chromosomal abnormalities was 31.6% (78/247), which included 66 cases with chromosomal aneuploidies and 12 with copy number variants (CNVs). CNVs accounted for 31.4% (11/35) of total abnormalities among fetuses with isolated increased NT, whilst only 2.3% (1/43) of the total abnormalities among fetuses with non-isolated increased NT. Three fetuses with a normal CMA result had mental and physical retardation. Two of them were diagnosed with single gene disorders by whole exome sequencing. CONCLUSION CMA can detect more chromosomal microdeletion/microduplications among fetuses with isolated increased NT. Furthermore, fetuses with increased NT and anegative CMA result during pregnancy cannot exclude all adverse outcomes.
Aneuploidy ; Chromosome Aberrations ; Chromosomes ; DNA Copy Number Variations ; Edema ; Female ; Fetus ; Humans ; Lymphangioma, Cystic ; Microarray Analysis ; Nuchal Translucency Measurement ; Pregnancy ; Pregnancy Outcome ; Prenatal Diagnosis ; Ultrasonography, Prenatal

Objective:To retrieve relevant evidence on exercise assessment and exercise training for children with congenital heart disease at home and abroad, and to summarize the best evidence to provide reference for clinical medical staff.Methods:UpToDate, National Guidelines Clearinghouse (NGC), Registered Nurses Association of Ontario (RNAO), Scottish Intercollegiate Guidelines Network (SIGN), National Institute for Health and Care Excellence (NICE), BMJ Best Practice, American Heart Association (AHA), Cochrane Library, Joanna Briggs Institute(JBI) Evidence-Based Health Care Center Database, PubMed, CINAHL, China Biomedical Literature Database, Yimaitong, Wangfang Database, CNKI were searched, related evidence on exercise assessment and exercise training for children with congenital heart disease. The search period was from the establishment of the database to March 2021. Clinical decision-making and recommended practice used retrospective evaluation methods for quality evaluation; guidelines used the 2012 version of the clinical guideline research and evaluation system (AGREE Ⅱ) for evaluation; systematic reviews used the systematic evaluation tool (AMSTAR) for evaluation; expert consensus used JBI (2016 version) evaluate the authenticity evaluation tools of opinions and consensus articles. Two researchers independently evaluated the literature, combined with the judgment of professionals, and extracted the literature data that met the standards.Results:A total of 15 documents were included, including 2 clinical decisions, 4 guidelines, 1 recommended practice, 5 systematic reviews, 3 expert consensus, and 22 best evidences. Including related personnel, exercise evaluation, exercise monitoring, exercise classification, exercise training, and exercise follow-up.Conclusions:This study summarizes the best evidence of exercise assessment and exercise training for children with congenital heart disease, and provides evidence-based evidence for clinical practice. It is recommended that children with congenital heart disease undergo exercise assessment and formulate a personalized exercise training program to promote the transformation of the best evidence into clinical practice.