Objective To evaluate the effect of electric vagal stimulation on myocardial ischemia-reperfu-sion (I/R) injury in rats.Methods Forty-five male Sprague-Dawley rats,aged 4-5 months,weighing 180-220 g,were randomly divided into 3 groups (n =15 each):sham operation group (S group),I/R group,electric vagal stimulation group (EVS group).Myocardial I/R was induced by occlusion of left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion in I/R and EVS groups.In group EVS,the left cervical vagus nerve trank was stimulated for 30 min with continuous electric pulses (5 V,2 ms,10 Hz) starting from 10 min before reperfusion until 10 min of reperfusion.The animals were sacrificed at the end of 120 min reperfusion and the hearts were removed for determination of the infarct size,TNF-α and IL-6 contents and myeloperoxidase (MPO) activity in myocardial tissues.Results Compared with group S,the infarct size was significantly enlarged,and TNF-α and IL-6 contents and MPO activity were increased in I/R and EVS groups (P ＜ 0.05).The infarct size was significantly smaller,TNF-α and IL-6 contents and MPO activity were lower in EVS group than in I/R group (P ＜ 0.05).Conclusion Electric vagal stimulation can reduce myocardial I/R injury in rats and activation of cholinergic anti-inflammatory pathway is involved in the mechanism.

Objective To investigate the difference among neonatal rat models of hypoxia-ischemia induced white matter injury of brain by three different methods and to provide theory basis on choosing the proper animal model for deep study on white matter injury of premature infants.Methods Three different hypoxia-ischemia methods were applied to induce brain white matter injury model of Wistar rats of postnatal 3rd day which were left carotid artery ligation followed by 6％ oxygen for 30 minutes and for 4 hours respectively,and bilateral carotid artery ligation followed by 8％ oxygen for 30 minutes.Compared brain pathological results of rats in different groups,changes in white matter of brain by glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) immunostaining,and changes in time of opening eyes and eyes appearance.Results Left carotid artery ligation followed by 6％ oxygen for 30 minutes resulted in sparse and vague periventricular white matter without necrotic lesions,and the degree of GFAP intensity [(5 021.63 ± 358.92) OD] increase and MBP intensity [(18 488.63 ± 1 822.62) OD] decrease were lowest (P ＜ 0.000 1),as well as the delay of opening time of the left eye.Left carotid artery ligation followed by 6％ oxygen for 4 hours resulted in necrotic lesions of periventricular white matter,with the degree of GFAP intensity [(6 069.13 ± 458.61) OD] and MBP intensity [(15 003.38 ± 1 559.11) OD],and also the delay of opening time of the left eye.Bilateral carotid artery ligation followed by 8％ oxygen for 30 minutes caused cystic necrosis of bilateral periventricular white matters,and the degree of GFAP intensity [(6 194.50 ±432.69) OD] increase and MBP intensity [(10 119.35 ± 735.16) OD] decrease of the left side were highest(P ＜ 0.000 1),as well as the delay of opening time and cataract in both eyes.Conclusion Left carotid artery ligation followed by 6％ oxygen for 30 minutes is more suitable for investigating mild brain white matter injury.Left carotid artery ligation followed by 6％ oxygen for 4 hours and bilateral carotid artery ligation followed by 8％ oxygen for 30 minutes are more proper for the investigation of severe periventricular leukemacia cases.

Objective To compare HER-2 state in breast cancer tissue deteced by fluorescent in situ hybridization (FISH) and immunohistochemistry (IHC) and analyze their correlation. Methods HER-2/neu protein expression and gene amplification were detected by FISH and IHC in 56 newly-diagnosed cases of female breast cancer from July 2008 to July 2009. Results Of the 56 patients, HER-2 protein expression (-), (+), (++), (+++) was 9 cases (16.1%), 29 cases (51.8%), 11 cases (19.6%) and 7cases (12.5%) respectively; 26 cases (46.4%) had HER-2 gene amplification while 30 cases (53.6%) didnt have. Type of HER-2 gene amplification was mainly HER-2(++) and HER-2(+++), and according gene amplification rate was 72 7% and 100%. HER-2 (+) gene amplification rate was 37.9 %(11cases) and no gene amplification was found in HER-2(-) tissue. The HER-2 positive rate using two methods had significant difference(χ2=19.778,P<0.01). HER-2(-) and HER-2(+++) had good consistency with the FISH results(Kappa=0.969),but HER-2(+) and HER-2(+ +) were poorly consistent with the FISH results(Kappa=0.271). Conclusions IHC is the preliminary screening method for detection of HER-2 expression. HER-2(-) and HER-2(+++) have good consistency with the gene amplification, and can guide clinical treatment. Some patients with HER-2(+) and HER-2(++) have HER-2 gene amplification. FISH is needed for targeted therapy.

Objective To evaluate the application value of bedside lung ultrasound in diagnosis of neonatal severe pneumonia.Methods Bedside lung ultrasound and chest X-ray were performed in 59 the newborn patients admitted to the newborn department for suspected severe pneumonia,from October 2015 to March 2016.According to the characteristics of the chest X-ray,patients were divided into consolidation group (widely spot shadow in double lung and airbronchograms) and the unconsolidation group (enlargement,fuzzy texture in double lung or small patch sample changes) Differences in ultrasonic characteristics were compared between the two groups.Results Ultrasonic image characteristics were pulmonary consolidation with air bronchograms,A-line disappearance,pleural line abnormalities.Among 23 patients with chest X-ray,21 patients were found pulmonary consolidation by lung ultrasound,and all 36 patients in unconsolidation group were not found pulmonary consolidation by lung ultrasound,and there was no significant difference in the detection rate by lung ultrasound between the two groups (P＞0.05) The pulmonary ultrasound results of 20 cases in the consolidation group showed A-line disappearance,37 cases in the unconsolidation group showed A-line disappearance.There was no statistical significant differences between the two groups (P＞0.05).The pulmonary ultrasound results showed 11 cases in the consolidation group and 15 cases in the unconsolidation group with pleural line abnormalities,there was no statistically significant differences between the two groups (P＞O.05).Conclusions Ultrasound image characteristics of neonatal severe pneumonia include pulmonary consolidation and air bronchograms,A-line disappearance,pleural line abnormalities.Compared lung ultrasound and chest X-ray result,there was no difference between consolidation group and unconsolidation group.Both of the two methods can be used for evaluating the diagnosis of pneumonia.

Objective To observe the clinical efficacy and adverse reaction of Chuanxiongqingnaokeli for children with migraine. Methods One hundred children patients with migraine were randomly divided into treatment group( n = 50 ) and control group( n = 50 ). Patients in treatment group were given Chuanxiongqingnaokeli,10 g/once,and three times one day,while in control group were given Flunarizine Hydrochloride Capsules,2. 5 mg/once,and who's body mass ﹥50 kg with 5. 0 mg/once,and one times each night. Three months as one course of treatment,and compared the efficacy of two groups after tree course of treatment. Results The hemodynamics of two groups all decreased after treatment compared with before treatment,but ACA,MCA,PCABA and VA in treatment group(( 81. 10 ± 11. 95 ),( 93. 3 ± 14. 16 ),( 70. 2 ± 11. 57),(70. 6 ± 13. 02),(65. 5 ± 12. 6)cm/s respectively)decreased more significantly than that of control group(( 104. 2 ± 12. 63 ),( 116. 2 ± 15. 82 ),( 93. 5 ± 11. 91 ),( 93. 5 ± 12. 71 ),87. 4 ± 12. 92 ) cm/s respectively),and the differences were significant( P﹤0. 05). The headache frequency and duration in treatment group were(1. 0 ± 0. 6)and(3. 3 ± 1. 0),less than that of control group((2. 3 ± 0. 9)and(5. 6 ± 1. 7);t= -3. 345,-3. 269;P﹤0. 05). The total effective rate in treatment group was 90. 0%(45/50),higher than that of control group(74. 0%(37/50);χ2 =4. 336,P﹤0. 05). There was no severe adverse reaction in both two groups. Conclusion The Chuanxiongqingnaokeli is safe and effective for treatment of children with migraine.

Objective To establish a dynamic model of lipopolysaccharide-induced acute lung injury in mice based on the NLRP3/Caspase-1/gasdermin D(GSDMD)pyroptosis pathway,and observe the result ing lung injury at different time points.We aimed to identify the optimal time for modelling according to the injury at different time points and the expression of pyroptosis pathway-related proteins,to lay the foundation for animal models for subsequent experiments.Methods Fifty-four 6～8 weeks old male SPF BALB/c mice were divided randomly into nine groups,including Con group and model groups at 1,3,6,12,18,24,48,and 72 h.Body weight and lung tissue were detected by general and pathological observations and semi-quantitative scoring,including lung index,lung water content,and wet and dry weight ratio.The white blood cell count and concentrations of tumor necrosis factor-α,interleukin(IL)-6,IL-1β,IL-18,and BCA protein were detected in bronchoalveolar lavage fluid(BALF).The classic pyroptosis pathway-related proteins NLRP3,pro-Caspase 1,Caspase 1,and GSDMD were detected by Western Blot.Results Body weight decreased in all experimental groups,with the most significant weight loss in the 24 and 48 h groups.Gross observation and pathological examination of lung tissue showed that the most severe lung injury occurred at 24～72 h,with significant differences between each group and the control group.The lung index,lung water content,and wet/dry weight ratio were also significantly increased at 24～72 h.White blood cells in BALF started to increase from 6 h after model initiation,48 h can reach a peak,72 h all keep increasing.IL-18 in BALF began to increase at 24 h and continued to increase at 72 h.The inflammatory factors tumor necrosis factor-α,IL-1β,IL-6 were highest at 6 h and significantly reduced at 48 h.Protein concentrations in BALF were significantly increased within 24,48,and 72 h compared with those in the control group.The pyroptosis pathway proteins NLRP3,pro-Caspase-1,Caspase-1,and GSDMD were significantly enhanced in each time series,and channel protein expression was significantly enhanced at 24～72 h compared with that in the Con group.Conclusions Comprehensive analysis of experimental indicators,inflammatory factors,and pathway proteins at different times showed that the mechanism of pyroptosis was closely related to the occurrence and progression of acute lung injury.Expression of the pyroptosis pathway was most obvious and lung injury was most serious at 24～48 h.This study provides a model reference and experimental basis for subsequent studies of the specific mechanism and intervention targets of acute lung injury.

Objective:To investigate the correlation between the expression of cyclic RNA PIP5K1A (circPIP5K1A) and microRNA-124-3p (miR-124-3p) in the serum and the severity of the disease and infertility in patients with endometriosis (EM) .Methods:A total of 110 cases of EM patients admitted to Shanxi Provincial Children's Hospital (Shanxi Maternal and Child Health Hospital) from Jan. 2020 to Jan. 2022 were regarded as the research objects, and were diveded into infertility group (44 cases) and successful pregnancy group (66 cases) according to the pregnancy outcome. General data were collected and the levels of serum miR-124-3p and circPIP5K1A were compared between the two groups; Pearson correlation was applied to analyze the correlation between serum miR-124-3p and circPIP5K1A. The diagnostic value of miR-124-3p and circPIP5K1A levels in patients with stage III-V EM was evaluated by ROC curve; Logistic regression analysis was applied to analyze the factors influencing the occurrence of infertility.Results:The expression level of serum miR-124-3p in stage Ⅰ EM patients (1.06±0.21) was higher than that in stage Ⅱ (0.83±0.19), stage Ⅲ (0.65±0.12) and stage Ⅴ (0.44±0.08), and the serum miR-124-3p level decreased with the increase of r-AFS stage. The differences were statistically significant ( t=54.29, P<0.05). The expression level of circPIP5K1A (0.77±0.12) was lower than that of stage Ⅱ (0.90±0.15), stage Ⅲ (1.20±0.20) and stage Ⅴ (1.53±0.26), and increased with the increase of r-AFS stage, and the differences were statistically significant ( t=90.45, P<0.05). Pearson correlation analysis showed that the expression of circPIP5K1A and miR-124-3p in serum of EM patients was negatively correlated ( r=-0.424, P<0.05) ; the serum miR-124-3p level in infertility group was lower than that in successful pregnancy group, and the serum circPIP5K1A level in infertility group was higher than that in successful pregnancy group ( P<0.05). ROC curve results showed that the area under the curve (AUC) of serum miR-124-3p and circPIP5K1A levels in the diagnosis of stage III-V EM patients was 0.900 and 0.887 respectively. The corresponding sensitivity was 88.24%, 85.29%, and the specificity was 78.95%, 88.16%, respectively. The AUC of combined diagnosis was 0.937, the sensitivity was 82.35%, and the specificity was 90.79%. Multivariate Logistic regression analysis showed that miR-124-3p and circPIP5K1A were related factors affecting infertility ( P<0.05) . Conclusions:The expression of miR-124-3p in the serum of EM patients decreases, and the expression of circPIP5K1A increases. The combined detection of miR-124-3p and circPIP5K1A has certain diagnostic value for the patients with stage III-V EM, and they are closely related to the occurrence of infertility.

Alfalfa(Medicago sativa L.)is the most important legume forage crop worldwide with high nutritional value and yield.For a long time,the breeding of alfalfa was hampered by lacking reliable information on the autotetraploid genome and molecular markers linked to important agro-nomic traits.We herein reported the de novo assembly of the allele-aware chromosome-level genome of Zhongmu-4,a cultivar widely cultivated in China,and a comprehensive database of genomic variations based on resequencing of 220 germplasms.Approximate 2.74 Gb contigs(N50 of 2.06 Mb),accounting for 88.39％of the estimated genome,were assembled,and 2.56 Gb contigs were anchored to 32 pseudo-chromosomes.A total of 34,922 allelic genes were identified from the allele-aware genome.We observed the expansion of gene families,especially those related to the nitrogen metabolism,and the increase of repetitive elements including transposable elements,which probably resulted in the increase of Zhongmu-4 genome compared with Medicago truncatula.Population structure analysis revealed that the accessions from Asia and South America had rela-tively lower genetic diversity than those from Europe,suggesting that geography may influence alfalfa genetic divergence during local adaption.Genome-wide association studies identified 101 sin-gle nucleotide polymorphisms(SNPs)associated with 27 agronomic traits.Two candidate genes were predicted to be correlated with fall dormancy and salt response.We believe that the allele-aware chromosome-level genome sequence of Zhongmu-4 combined with the resequencing data of the diverse alfalfa germplasms will facilitate genetic research and genomics-assisted breeding in variety improvement of alfalfa.

Objective: To prepare peptide minotope-based recombinant diagnostic antigen of Epstein-Barr virus (EBV) infection and evaluate its antigenicity preliminarily. Methods: With Trx at the N-terminal and His tag at the C-terminal, the peptide minotope of EBV (GP125, F1, A2, A3C2) was expressed in Escherichia coli and purified by affinity and anion exchange chromatography (designated 'H58’); based on antigenicity of H58 identified by Western blotting (WB), we constructed and evaluated a novel early diagnostic ELISA for EBV infection. Results: The soluble H58 protein with high concentration (2.8 mg/ml) and purity (99.01) was obtained; WB analysis found that there was an obvious band (28 ×10³) on the NC membrane, using H58 anti-Trx monoclonal antibody or acute-phase sera of EBV infection as the first antibody. With the novel ELISA, 50 positive sera of EBV infection and 50 negative sera were detected, displaying that the grouping of OD value of positive serum (95%CI: 1.233-1.489) and negative serum (95%CI: 0.113-0.159) was different (P<0.05) with the sensitivity 98.0%, specificity 96.0% and kappa value 0.940. Conclusions By E. coli expression and affinity and ion exchange chromatography purification, the peptide minotope-based recombinant diagnostic antigen of EBV infection was obtained with excellent antigenicity, which could be applied for serological detection of EBV infection.

Antrodia cinnamomea is extensively used as a traditional medicine to prevention and treatment of liver cancer. However, its comprehensive chemical fingerprint is uncertain, and the mechanisms, especially the potential therapeutic target for anti-hepatocellular carcinoma (HCC) are still unclear. Using UPLC‒Q-TOF/MS, 139 chemical components were identified in A. cinnamomea dropping pills (ACDPs). Based on these chemical components, network pharmacology demonstrated that the targets of active components were significantly enriched in the pathways in cancer, which were closely related with cell proliferation regulation. Next, HCC data was downloaded from Gene Expression Omnibus database (GEO). The Cancer Genome Atlas (TCGA) and DisGeNET were analyzed by bioinformatics, and 79 biomarkers were obtained. Furtherly, nine targets of ACDP active components were revealed, and they were significantly enriched in PI3K/AKT and cell cycle signaling pathways. The affinity between these targets and their corresponding active ingredients was predicted by molecular docking. Finally, in vivo and in vitro experiments showed that ACDPs could reduce the activity of PI3K/AKT signaling pathway and downregulate the expression of cell cycle-related proteins, contributing to the decreased growth of liver cancer. Altogether, PI3K/AKT-cell cycle appears as the significant central node in anti-liver cancer of A. Cinnamomea.