The aim of this study was to investigate the effects of high-intensity interval training(HIIT)on lipopolysaccharide(LPS)-induced septic myocardial injury in mice and the roles of NLRP3 inflammasome and macrophage M1 polarization in the process.C57BL/6 male mice were randomly divided into 4 groups:control(CON)group,LPS(L)group,HIIT+saline injection(E)group,and HIIT+LPS(EL)group.Six weeks of HIIT intervention was followed by intraperitoneal injection of LPS,and cardiac function indexes were measured by echocardiography 12 hours post the injection.Hematoxylin-eosin(HE)staining was used to evaluate the morphology and pathological characteristics of myocardium for assessing myocardial damage score;enzyme-linked immunosorbent assay(ELISA)was used to test the content of myocardial damage indicators(AST,CK-MB,LDH);RT-PCR was used to detect the relative mRNA levels of NLRP3 inflammasome(NLRP3,Caspase-1),atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),myeloperoxidase(MPO)and macrophage M1-associated inflammasome factors(IL-1β,TNF-α,IL-6);Western blot was applied to measure the protein expression of inducible nitric oxide synthase(iNOS)and apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC)in cardiac tissues;immunofluorescence staining was used to detect the protein expression of NLRP3 inflammasome,ASC,IL-18 and iNOS.Compared with the CON group,mice in the LPS group showed obvious decrease in body weight,a significant decrease in EF and FS,a significant increase in LVESD and LVEDD,obvious pathological damage in myocardial tissue,a significant increase in myocardial damage fraction,a significant increase in serum myocardial damage indexes,and a significant increase in the expression levels of BNP,MPO,NLRP3 inflammasome,iNOS,IL-1β,IL-6 and TNF-α.HIIT treatment could reverse these changes mentioned above in model mice.In conclusion,6 weeks of HIIT inhibits the activation of LPS-induced NLRP3 inflammasome and suppressed macrophage M1-type polarization,thereby combating septic myocardial injury.

Objective:To investigate HER2 mRNA expression in breast cancer with HER2 immunohistochemistry (IHC) 0 and to analyze the feasibility of distinguishing between the tumor with HER2 μltra-low expression and the one without expression of HER2 (no staining by IHC) by HER2 mRNA level preliminarily.Methods:HER2 mRNA was analyzed by reverse transcription digital PCR in 41 cases of formalin-fixed paraffin-embedded surgical tissue samples of invasive breast cancer obtained between January 2020 and March 2023 at Peking Union Medical College Hospital. The cohort included 21 HER2 IHC 1+ and 20 IHC 0 (12 ultra-low and 8 non-expression of HER2). HER2 mRNA expression level was quantitatively evaluated by the FAM (HER2)/VIC (reference gene) ratio.Results:The expression of HER2 mRNA for the cases with 1+, ultra-low, and non-expression of HER2 by IHC was 0.30 to 1.78 (average 0.90, median 0.82), 0.55 to 1.51 (average 0.93, median 0.90) and 0.22 to 0.78 (average 0.41, median 0.36), respectively. For the mean and median HER2 mRNA levels, there was no significant difference between HER2 IHC 1+ and HER2 ultra-low expression diseases ( P=0.757). A remarkable difference in HER2 gene expression was found between the tumors with 1+ and non-expression of HER2 by IHC ( P=0.002). And, HER2 ultra-low cases contained statistically higher levels of HER2 mRNA compared with non-expression of HER2 subgroup by IHC ( P=0.001). Conclusions:Based on HER2 mRNA, HER2 non-expression and HER2 weak expression (including HER2 IHC 1+ and ultra-low) belong to two different types of the tumor and the disease with HER2 IHC 1+ and HER2 ultra-low expression may be the same. It is necessary to further test the performance of HER2 mRNA detection for stratifying the HER2 weak expression subgroup and to determine the threshold.

Objective:To investigate the clinical and pathological characteristics of breast cancer with HER2 low expression.Methods:The data from 3 422 patients with invasive breast cancer which archived in Peking Union Medical College Hospital between April 2019 and July 2022 were retrospectively reviewed. Among them, 136 patients were treated with neoadjuvant chemotherapy. The tumor size, histological type, tumor differentiation, lymph node metastasis, Ki-67 index, the status of estrogen receptor, progesterone receptor and HER2 as well as pathological complete response (pCR) rate were collected.Results:The HER2 status of 3 286 patients without neoadjuvant therapy, 616 (616/3 286, 18.7%) score 0, 1 047 (1 047/3 286, 31.9%) score 1+, 1 099 (1 099/3 286,33.4%) score 2+ and 524 (524/3 286,15.9%) score 3+ by immunohistochemistry (IHC). Among the 1 070 IHC 2+ cases, 161 were classified as HER2 positive by reflex fluorescence in situ hybridization (FISH) assay. In our cohort, 1 956 cases of HER2-low (IHC 1+ and IHC 2+/FISH-) breast cancer were identified. Compared to the HER2 IHC 0 group, HER2-low tumors more frequently occurred in patients with hormone receptor (HR) positive ( P<0.001), Ki-67 index below 35% ( P<0.001), well or moderate differentiation ( P<0.001) and over the age of 50 ( P=0.008). However, there were no significant differences in histological type, tumor size, and lymph node metastasis between HER2-low and HER2 IHC 0 group. For patients who had neoadjuvant therapy, the pCR rate in the patients with HER2-low was lower than those with HER2 IHC 0 (13.3%, 23.9%), but there was no significant difference. Although HER2-low breast cancers showed a slightly lower pCR rate than HER2 IHC 0 tumors, no remarkable difference was observed between tumors with HER2-low and HER2 IHC 0 regardless of hormone receptor status. Conclusions:The clinicopathological features of HER2-low breast cancers are different from those with HER2 IHC 0. It is necessary to accurately distinguish HER2-low breast cancer from HER2 IHC 0 and to reveal whether HER2-low tumor is a distinct biological entity.

The prevention and treatment of cardiovascular diseases have always been a focus of attention in re-lated fields.This paper discusses the application and mechanism of exogenous and endogenous supplementation with β-hy-droxybutyrate in the prevention and treatment of cardiovascular diseases such as myocardial ischemia,myocardial infarc-tion,diabetic cardiomyopathy,hypertension,myocardial inflammation,hypertrophic cardiomyopathy and heart failure.Exogenous β-hydroxybutyrate supplementation can quickly and directly provide substitute energy for the heart and thus prevent and treat heart injury.The comprehensive effect of exogenous β-hydroxybutyrate supplementation is significantly better than that of endogenous β-hydroxybutyrate.Endogenous fatty acid,obtained through a ketogenic diet,needs to be oxidized to β-hydroxybutyrate in vivo for energy supply,which is greatly influenced by glucose and lipid metabolism under the conditions of the basic disease itself.At present,ketogenic diet is not recommended for use in the prevention and treat-ment of myocardial ischemia or myocardial infarction,and in the treatment of diabetic cardiomyopathy or hypertension.

Ketogenic diet can supplement blood ketone,provide energy for skeletal muscle,and affect the athletic performance and recovery of animals and humans after fatigue.In this paper,it was found that ketogenic diet transformed muscle fibers from Ⅱb to Ⅱa through axonal germination and nerve reinnervation,improved the quality and function of mitochondria of fast muscle and increased histone acetyltransferase to enhance skeletal muscle strength.Using ketone body to provide energy,it can reduce glycolysis and improve the ability of fatty acid oxidation in slow muscle to improve exercise endurance;It can reduce endoplasmic reticulum stress,oxidative stress and inflammatory reaction of skeletal muscle,protect the body from injury,reduce the consumption of muscle glycogen and the accumulation of lactic acid,reduce fatigue after exercise and promote recovery.It is considered that ketogenic diet has low negative effects on the body,can improve sports performance and accelerate fatigue recovery,and can be used as a diet scheme in competitive sports and rehabilitation training.

Identification of genetic variants via high-throughput sequencing (HTS) technologies has been essential for both fundamental and clinical studies. However, to what extent the genome sequence composition affects variant calling remains unclear. In this study, we identified 63,897 multi-copy sequences (MCSs) with a minimum length of 300 bp, each of which occurs at least twice in the human genome. The 151,749 genomic loci (multi-copy regions, or MCRs) harboring these MCSs account for 1.98％of the genome and are distributed unevenly across chromosomes. MCRs containing the same MCS tend to be located on the same chromosome. Gene Ontology (GO) anal-yses revealed that 3800 genes whose UTRs or exons overlap with MCRs are enriched for Golgi-related cellular component terms and various enzymatic activities in the GO biological function cat-egory. MCRs are also enriched for loci that are sensitive to neocarzinostatin-induced double-strand breaks. Moreover, genetic variants discovered by genome-wide association studies and recorded indbSNP are significantly underrepresented in MCRs. Using simulated HTS datasets, we show that false variant discovery rates are significantly higher in MCRs than in other genomic regions. These results suggest that extra caution must be taken when identifying genetic variants in the MCRs via HTS technologies.

Background Oxidative stress is a main cause of age-related macular degeneration (AMD).Lutein has a preventive role for AMD, but its antioxidant mechanism remains unclear.Objective Present study was to investigate the effect of lutein on oxidative stress of Müller cells and its signaling pathway.Methods Human Müller cells (human Müller cell strain) were cultured, and the cells at logarithmimic growth phase were incubated in 96 well plate overnightly.Oxidative stress cell models were established by adding 160 μmol/L H2O2, a median lethal dose for Müller cells.The models were divided into the model control group and 12.5,25.0,50.0 mg/L lutein groups,and the different concentrations of lutein were used to culture the cells for 24 hours, respectively.The routine cultured cells served as the blank control group.Growth of the cells was assayed by MTT method (absorbancy);the reactive oxygen species (ROS) content in the cells was assayed by flow cytometry;the mRNA and protein levels of nuclear factor-E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in the cells were detected by quantitative real-time PCR and Western blot, respectively.Results The inhibitory effects on the cells were gradually enhanced with the increase of H2O2 concentrations,showing a significant difference among the groups (F =43.890,P＜0.01).A significant difference was found in apoptotic rate of the cells among the blank control group,model control group and 12.5,25.0,50.0 mg/L lutein groups (F =346.770, P =0.000) , and the apoptosis rate was significant elevated with the increase of lutein dose (all at P＜0.05).The ROS contents in the cells were 1.92±0.18,64.89±2.86,52.70±2.80,32.61 ±4.20 and 5.68 ± 1.35 in the blank control group, model control group and 12.5,25.0,50.0 mg/L group, respectively, with significant difference among the groups (F =324.900, P =0.000), and the ROS content was gradually reduced as the increase of lutein dose (all at P＜0.05).The relative mRNA and protein expressions of Nrf2 and HO-1 were remarkedly higher in the 12.5,25.0,50.0 mg/L lutein groups than those in the model control group (F =236.960,242.620,186.830,263.120, all at P =0.000) , and no significant difference was seen in the relative expression level of nuclear Nrf2 protein among the groups (F =1.790, P =0.210).Conclusions Lutein can induce the expression of antioxidant enzymes by inducing the expression of nuclear translocation of Nrf2 and consequently inhibit the oxidative stress status.

Objective To reveal the effect of lutein on the status of oxidative stress in rats with high homocysteine levels (HHcy) and relevent molecular mechanisms.Methods The wistar rat HHcy model was established by intra-gastric administration with L-methionine suspension and treated with lutein.The oxidative stress status and the gene expression changes of transcription Nf-E2-related factor2 (Nrf2),a regulation factor of down stream antioxidant protein gene expression,were detected in HHcy rats and lutein intervention rats.Results Compared with the rat serum SOD activities (134.32 ± 12.65) U/mL in the control group,the serum SOD activities in the HHcy model group (95.6 ± 10.92) U/mL were significantly lower (P ＜ 0.05).The serum glutathione peroxidase (GPx) activities in the HHcy model group (121.66 ± 18.64) U/mL were also significantly lower as compared with the the control group (183.17 ± 21.29) U/mL,P ＜ 0.05.However,the serum SOD (126.75 ± 11.26) U/mL and GPx activities (167.18 ± 19.66) U/mL in the lutein intervention group were significantly higher as compared with the HHcy model group.As compared with the rat serum malondialdehyde (MDA) content (5.11 ± 0.68) μmol/L as well as the hydroxyl free radical levels (0.53 ± 0.05) U/L in the control group,the serum MDA content (7.65 ± 0.87) μmol/L and the hydroxyl free radical levels (0.92 ± 0.09) U/L in the HHcy model group were significantly higher.The serum MDA content (6.44 ±0.91) μmol/L and the hydroxyl free radical levels (0.74 ± 0.06) U/L in the lutein intervention group were significantly lower as compared with the HHcy model group.RT-PCR and Western blot results also showed decreased expression of SOD2 and GPxl mRNA in aorta epithelial tissues of HHcy model rats.With lutein intervention,the expressions ofSOD2 and GPx1mRNA were significantly increased and the gene expression of Nrf2 also up-regulated.Conclusions The Hhcy model rats were under the status of augmented oxidative stress,and carotenoid lutein could attenuate the Hcy-mediated oxidative stress,and its mechanism might be potentially associated with up-regulating the expression of Nrf2,thereby inducing the expression of its downstream antioxidant proteins.

Objective To revise the standards of the Language Function Evaluation for Hearing-impaired Children and to study the characteristics of language functions in hearing-impaired children.Methods A total of 741 normal hearing children at the age of 3~7 year old from 15 provinces in China were assessed using the Language Function Evaluation for Hearing-impaired Children.A total of 90 hearing-impaired children also completed the e-valuation.The scores of Representation Function,Tool Function,Coordination Function,Expression Function, Entertainment Function and Comprehensive Function of all the children were collected.ResuIts The regression e-quations of the language function were established:Representation Function score=3.34 +0.81 ×children's age, Tool Function score =6.00+0.51×children's age,Coordination Function score =5.72 +0.52 ×children's age,Ex-pression Function score =4.44+0.67×children's age,Entertainment Function score =6.34+0.47×children's age, Comprehensive Function score =4.96+0.62×children's age;Through the regression analysis,the reference stand-ards of the tool for children at the age of 3~6 year old were set down.Five-year-old hearing-impaired children's Tool Function and Expression Function were better than those of three or four-year-old ones(P<0.05).Five-year-old hearing-impaired children's Comprehensive Function were better than those of three-year-old ones(P<0.05).The language function scores had a big gap between the hearing-impaired children and the hearing-nor-mal children at the same ages.ConcIusion The new reference standards of the Language Function Evaluation for Hearing-impaired Children provides the Expectation values and Minimum values of language function from children at the age of 3 -6.They are rehabilitation criterions for hearing-impaired children.The older the hearing-im-paired children are,the higher their scores of language function evaluation.But their scores are much lower than those of the normal hearing children at the same age.

Objective To study the characteristics of language function development and establish the reference standards in children of4~6 years old. Methods 616 hearing normal children at the age of 3~7 from 13 provinces in China were assessed with Language FunctionEvaluation for Hearing-impaired Children. 583 usable data were analyzed via SPSS 17.0. Results The older the children were, the highertheir scores of language function evaluation were (P<0.001). The differences between boys and girls on coordination function and relaxationfunction were significant (P<0.01). The girls were better than the boys. The reference standards of children of 4~6 years old were set downbased on the regression analysis. Conclusion There are some laws on children's language function development. The reference standards ofchildren by 4~6 years old has been developed.