BACKGROUND: Doxorubicin hydrochloride (DOX) is extensively used in the treatment of various tumors. However, its clinical application is limited due to dose-dependent cardiotoxicity. Currently, few effective strategies exist to mitigate or eliminate DOX-induced cardiomyopathy (DIC). Although ferroptosis is implicated in DIC and its inhibition partially alleviates the condition, the direct targets of DOX in the progression of cardiotoxicity remain unclear. This study aimed to discover the direct targets of DOX in ferroptosis-mediated DIC. METHODS: A DOX pulldown assay was performed to identify proteins specifically binding to DOX in murine hearts, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify candidate proteins. A cardiac injury mouse model was established by DOX treatment. Based on this, multiple ferroptosis biomarkers were detected by flow cytometry, quantitative real-time polymerase chain reaction, western blotting, immunochemistry, etc. Besides, specific activator and inhibitor of signaling pathways were applied to illuminate molecular mechanisms. RESULTS: Glutathione S-transferase P1 (GSTP1) was identified as a DOX target. GSTP1 activity was inhibited in DOX-treated cardiomyocytes, while its overexpression significantly alleviated DIC. Moreover, GSTP1 overexpression inhibited acyl-CoA synthetase long-chain family member 4 (ACSL4)-dependent ferroptosis. Mechanistically, GSTP1 overexpression suppressed c-Jun N-terminal kinase (JNK) phosphorylation, thereby reducing reactive oxygen species (ROS) production and inhibiting ferroptosis in DIC. CONCLUSIONS This study identifies the DOX/GSTP1/JNK axis as a critical pathway mediating ACSL4-dependent ferroptosis in DIC. GSTP1 is highlighted as a potential key mediator of ferroptosis and a promising therapeutic target for DIC.

Breast cancer has become the malignant tumor with the highest incidence rate among women, of which human epidermal growth factor receptor 2 (HER2) positive breast cancer accounts for about 15%-20%. With the development of anti HER2 targeted drugs, the survival and prognosis of HER2 positive breast cancer has significantly benefited. About 45%-55% of breast cancer patients have low HER2 expression, and these patients usually do not receive anti HER2 treatment. However, there is a significant difference between the biological behavior and prognosis of breast cancer with low HER2 expression and breast cancer with zero HER2 expression. It is helpful to differentiate and adopt corresponding treatment strategies to improve the prognosis of patients. At present, there have been many advances in targeted therapy of breast cancer with low HER2 expression, which provides a useful reference for precision treatment of breast cancer with low HER2 expression.

Objective: To understand physical activity status and its relation with cardiopulmonary endurance among Chinese children and adolescents, so as to provide reference for cardiopulmonary endurance improvement and intervention of Chinese children and adolescents. Methods: During September to November 2018, a stratified cluster sampling method was used to select 4 269 children and adolescents aged 7-18 years in six major regions of China, 20 m shuttle run test and physical activity were assessed. Results: Average duration for daily lowintensity physical activity among boys was (31.49±30.01) min, for moderate to vigorous physical activity (68.37±46.50) min. Average duration for low intensity physical activity among girls was (34.02±31.58) min, for moderate to vigorous physical activity was (63.02±44.91) min. The number of 20 m shuttle run test for boys reached ( 36.17 ±21.94) times, and the maximal oxygen uptake per minute was (44.51±5.05) mL/kg; the number of 20 m shuttle run test for girls reached (26.59±12.56) times, and the maximal oxygen uptake per minute was (41.99±5.29) mL/kg. There were statistically significant differences in the number of 20 m shuttle run test and maximal oxygen uptake across age groups in boys ( F= 589.37 , 152.79, P <0.01), as well as that in girls ( F=305.21, 1 032.46, P <0.01). The proportion of boys and girls who met moderate to vigorous physical activity standard was 49.24% and 42.92%, respectively. Linear regression analysis showed that there was a significant positive correlation between the duration of moderate to vigorous physical activity and maximal oxygen uptake ( β=0.11, P ＜ 0.01). Conclusion There is a significant correlation between moderate to vigorous physical activity with cardiorespiratory endurance among Chinese children and adolescents. Higher level of moderate to vigorous physical activity is important to help improve cardiopulmonary endurance among children and adolescents.

Objective: To investigate the positive rate for 2019-nCoV tests and co-infections in Wuhan district. Methods: A total of 8 274 cases in Wuhan were enrolled in this cross-sectional study during January 20 to February 9, 2020, and were tested for 2019-nCoV using fluorescence quantitative PCR. Both respiratory tract samples (nasopharynx, oropharynx, sputum and alveolar lavage fluid) and non-respiratory tract samples (urine, feces, anal swabs, blood and conjunctival sac swabs) were collected. If both orf1ab and N genes are positive, they are classified as nucleic acid test positive group; if both orf1ab and N genes are negative, they are classified as negative group; if single gene target is positive, they are classified as suspicious group. Individuals were divided into male group and female group according to sex. At the same time, 316 patients were tested for 13 respiratory pathogens by multiplex PCR. Results: Among the 8 274 subjects, 2 745 (33.2%) were 2019-nCoV infected; 5 277 (63.8%) subjects showed negative results in the 2019-nCoV nucleic acid test; and 252 cases (3.05%) was not definitive (inconclusive result). The age of cases with COVID-19 patients and inconclusive cases was significantly higher than that of cases without 2019-nCoV infection (40 vs 56, t=27.569, P<0.001; 52 vs 56, t=6.774, P<0.001). The positive rate of 13 respiratory pathogens multiple tests was significantly lower in 104 subjects who were positive for 2019-nCoV compared with those in subjects who were negative for 2019-nCoV test (5.77% vs 18.39%, χ²=24.105, P=0.003). Four types of respiratory tract samples and five types of non-respiratory tract samples were found to be positive for 2019-nCoV nucleic acid test. Conclusion The 2019-nCoV nucleic acid positive rate in male is higher than in female. Co-infections should be pay close attention in COVID-19 patients. 2019-nCoV nucleic acid can be detected in non-respiratory tract samples.

We expressed B cell epitopes of dengue virus envelope protein and NS1 protein in prokaryotic cells,and purified and evaluated for its serological activities.A recombinant multi-epitope chimeric gene named rE including eight B cell epitopes was connected by linker peptide (EAAAK)2 and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The expressed recombinant protein was purified with 6× His purification media,and identified by SDS-PAGE and Western blot,and its antigenicity was analyzed by using an indirect ELISA assay.The recombinant expression vector pET28a-rE was constructed and expressed in BL21 (DE3) successfully,but the recombinant proteins mainly appeared as inclusion bodies.The target protein was obtained with high purity through the purification of affinity chromatography.SDS-PAGE and Western blot analysis showed that the molecular weight of fusion protein was in the expected line.The established indirect ELISA has high accuracy.This recombinant peptide antigen expressed in E.coli has good potential for serum testing.

Objective To evaluate the systolic function in patients with preeclampsia using three-dimensional speckle tracking imaging (3D-STI) technology.Methods Totally 73 patients with mild preeclampsia (mild group),64 patients with severe preeclampsia (severe group) and 60 healthy pregnant women with matched age and gestational weeks (control group) were enrolled in this study.All the biochemical indicators and conventional echocardiogram parameters were detected.The left ventricular global longitudinal systolic peak strain (GLS),global radial systolic peak strain (GRS),global circulation systolic peak strain (GCS) and global area systolic peak strain (GAS) were measured using 3D-STI,and the dyssynchronization index of area strain (ASDI) was calculated in all three groups.The correlation between sensitive three dimensional strain and biochemical indexes and echocardiogram parameters were analyzed.Results Compared with control group,the left atrial diameter (LAD) increased,E/A decreased in severe group,Tei index increased in turn at control group,mild group and severe group (all P＜0.05).Compared with control group,the |GLS| and |GAS| reduced and ASDI was increased in mild group,the changes were more significant in severe group.The GRS and GCS were also appeared lower in severe group (all P＜0.05).The correlation analysis showed a negative correlation between |GLS| and Tei index (r=-0.471,P=0.036),a positive correlation between | GAS| and LVEF (r=0.051,P=0.028),a negative correlation between | GAS| and Tei index (r=-0.612,P =0.017),a negative correlation between ASDI and H-FABP (r=-0.525,P=0.046),a positive correlation between ASDI and Tei index (r=0.489,P=0.037).Conclusion The myocardial strain capacity of left ventricular decrease and systolic function are significantly impaired in patients with preeclampsia.The 3D-STI technology can early noninvasively assess the change of left ventricular systolic function in patients with preeclampsia.

Objective:To establish a double antibody sandwich ELISA assay for detection of human IL-37 in serum.Methods: Mouse anti-human IL-37 monoclonal antibody was used as capturing antibody,rabbit anti-human IL-37 polyclonal antibodies served as detection antibody,HRP labeled goat anti-rabbit IgG employed as second antibody and recombinant human IL-37 protein used as reference standard for the establishment of a doubleantibody sandwich ELISA.The working conditions were optimized,such as sensitivity,linear range,reproducibility and evaluated the serum IL-37 in patients with Dengue fever.Results: The sensitivity of the established ELISA method was 1.465 μg/L approximately.Likewise,the linearity range of this method was about (1.465-46.875) μg/L.Further,the co efficient of variation (CV) of inter-batch and intra-batch in this study were 6.6％ and 11.7％,respectively.Notably,this method could be used in the detection of IL-37 in serum of the patients with Dengue fever,showing that the level of IL-37 in Dengue fever patients was much higher than that in healthy controls.Conclusion: The double antibody sandwich ELISA assay for the detection of human IL-37 was successfully established,which can be apply to detect of human IL-37 in clinical samples.

Objective To detect the genotype of hepatitis C virus(HCV)in chronic hepatitis C(CHC)infection patients using gene sequence method and observe the distributive characteristic of HCV genotype in Hubei.Methods A total of 447 HCV-RNA-positive plasma samples were collected from chronic hepatitis C patients in Infectious Diseases Department of Renmin Hospital of Wuhan University from February 2013 to July 2015.Then NS5B region gene sequence of HCV genome were de-tected by Sanger sequencing method and compared with HCV genotype in NCBI genebank database for analyzing HCV geno-type.Results A total of 11 kinds of genotypes were detected,including genotypes 1a,1b,2a,3a,3b,6a,6b,1b/2a,1b/2k,6a/1b and 6d/6k,respectively.Detection cases of various genotypes were respectively 7 cases (1.57%),325 cases (72.71%),67 cases (14.99%),7 cases (1.57%),20 cases (4.47%),14 cases (3.13%),2 cases (0.45%),2 cases (0.45%),1 case (0.22%),1 case (0.22%)and 1 case (0.22%).Conclusion Genotype 1b was the major type of HCV-RNA genotype,fol-lowed by 2a,also other genotypes existed,which prompted that the prevalence of HCV genotype was diversity in Hubei.

Objective To evaluate the accuracy and reliability of O-arm-based thoracolumbar pedicle screw installation.Methods A retrospective review was conducted on 75 patients who had undergone thoracolumbar pedicle screw fixation assisted with O-arm navigation (navigation group,n =32) and C-arm fluoroscopy (fluoroscopy group,n =43) from March to October 2014.All the patients were assessed with X-ray and CT images after operation.Accuracy of screw installation in both groups was compared.In navigation group,screw directions were measured on the sagittal and axial images of intraoperative navigation and post-operative CT scanning to evaluate the concordance.Results In fluoroscopy group,a total of 206 pedicle screws were placed with the one-time success rate of 93.2％ and accuracy of 90.8％ for screw placement,and one misplaced screw (grade llⅢ) led to L3 nerve root symptom.In navigation group,a total of 226 pedicle screws were placed with the one-time success rate of 100％ and accuracy of 96.9％ for screw placement,and no screw was grade Ⅲ.Further,there were no significant differences in screw directions on the sagittal and axial images between intraoperative navigation and postoperative CT scanning (P ＞ 0.05).Conclusion O-arm navigation that provides high-resolution images and high precision improves the accuracy of thoracolumbar pedicle screw installation,and possesses good reliability.

Objective To detect UGT1A1 gene promoter polymorphism with gene sequence method and analyze the distribu-tion of UGT1A1*28 and UGT1A1*6 gene polymorphism in Wuhan.Methods A total of 230 samples were collected from tumor patient in Medical Oncology Department of Renmin Hospital of Wuhan University from January 2013 to December 2014,then gene sequence of the target fragments were detected by Sanger sequencing method and analyzed patient UGT1A1 gene promoter polymorphism TA box.Results In this research examination 230 example tumor patient,198 patients (86.1%)were identified with TA6/TA6 genotype,29 patients (12.6%)were identified with TA6/TA7 genotype,3 pa-tients (1.3%)TA7/TA7 variant.Conclusion TA6/TA6 wild-typewas higher frequence in patients with malignant tumour in Wuhan.