Objective To establish a homozygous zebrafish map3k15 knockout line using CRISPR/Cas9 gene editing technology so to provide an animal model for investigation of potential role of map3k15 in renal diseases.Methods 1)Analyzing the expression pattern of map3k15 in zebrafish;2)Establishing a homozygous map3k15 knockout zebrafish line using CRISPR/Cas9;3)Observing the phenotypic effects of map3k15 deficiency in ze-brafish.Results 1)map3k15 was expressed in the zebrafish pronephros,and map3k15/map3k15 protein was found highly conserved across species;2)map3k15-/-mutant zebrafish model was constructed with+2 bp mutation and+1 bp mutation;3)map3k15 mutant zebrafish exhibited edema in yolk sac,pericardium and head during embryonic development.Conclusions A homozygous zebrafish map3k15 knockout line is successfully developed which may provide an important model for future research on the role of map3k15 in renal development and disease.

Objective To rvaluate the therapeutic efficacy of balloon-dilatation covered stents in the treatment of renal artery stenosis(RAS).Methods The clinical data of 30 patients with RAS,who received intravascular ultrasonography(IVUS)-guided LifeStream balloon-dilatation covered stent implantation at the Affiliated Central Hospital of Dalian University of Technology(Dalian Municipal Central Hospital)of China from August 2022 to December 2023,were retrospectively analyzed.The various parameters of the lumen and the stent were measured,and the performance of the stent was evaluated.Results The minimum original blood vessel diameter below the base of the stenotic segment plaque was 5.40(5.17,5.80)mm and the maximum blood vessel diameter was 6.20(5.80,6.93)mm,which became 6.00(5.80,6.00)mm and 7.90(6.00,8.00)mm respectively after stent release,the differences were statistically significant(both P＜0.05).Before stent release the luminal eccentricity index was(14.72±9.37)％,which was(1.54±9.16)％after stent release,the difference was statistically significant P＜0.05).The instant stent symmetry after stent release was(82.69±14.61)％,and the stent expansion factor was(99.81±10.70)％.Ideal narrow coverage rate was obtained.During operation,poor stent adhesion occurred in 2 patients and renal artery rupture with bleeding occurred in one patient,which were solved after immediate re-expansion treatment.Spearman's correlation analysis showed that stent symmetry,stent expansion factor,and stent eccentricity index did not linearly correlate with the lumen cross-sectional area stenosis rate and the plaque eccentricity index(all P＞0.05).Conclusion For the treatment of RAS,the LifeStream balloon-dilatation covered stent is clinically safe,feasible,and effective with satisfactory immediate clinical outcomes.

Objective:To investigate and analyze the status quo of work stress and its influencing factors among residents in the standardized training of obstetrics and gynecology.Methods:A questionnaire survey was carried out among 232 training residents by random sampling from a standardized training base of a tertiary hospital in Beijing from June 2019 to May 2020. The collected general information included gender, age, specialty, education background, work experience, time of training, income satisfaction and recognition of the necessity of training. The current situation and influencing factors of pressure were analyzed through the work pressure source scale. The lower the score means the greater stress. SPSS 22.0 was used to process the data by performing t-test and Chi-square test. Results:The total points of work stress (67.02±12.65) of the residents was lower among all the departments ( P<0.001). The most important work stressors were workload and time allocation (7.11±2.42), job situation and resources (7.21±2.34), management and personal relationship (15.66±3.69), and those were significantly different in any other departments ( P=0.003). One-way analysis of variances showed that job stress scores were significantly different among types of income satisfaction ( P=0.003). Combined with the results of logistic regression equation, the total scores of work stressors in such three aspects as "specialty and career", "workload and time allocation", and "job situation and resources" decreased with the satisfaction, and the lower the score, the greater the pressure, with statistical significance ( P=0.006, 0.008, 0.012, respectively). However, in terms of "patient diagnosis and treatment", and "management and interpersonal relationship", there was no significant difference between the groups ( P=0.067, 0.057, respectively). Conclusions:The stress of obstetrics and gynecology residents is generally high. Clinical and medical affairs are the main sources of their work stress, and income satisfaction is the major influencing factor of the work stress.

Airway mucus hypersecretion is one of the pathological changes in children with community acquired pneumonia（CAP），and affects the severity，progression and prognosis of CAP.Diagnosis of airway mucus hypersecretion currently relies on fiberoptic bronchoscopy. To predict the risk of airway mucus hypersecretion and to take early action to avoid serious consequences such as plastic bronchitis and hypoxia and respiratory failure，the article summarizes the relationship between age，clinical characteristics and laboratory indices and the formation of airway mucus hypersecretion in children with CAP. Among them，age，pathogenic species，fever peak and fever range，neutrophil ratio，C-reactive protein，lactate dehydrogenase，D-dimer，serum 25（OH）D，and some interleukins，interferons，and acquired immune indicators have an early warning effect on the development of airway mucus hypersecretion in children with CAP.

Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.

Gynecological diseases are one of the important factors that threaten women's reproductive health.Endoplasmic reticulum(ER)stress is a stress response induced by the accumulation of unfolded or misfolded proteins in ER cavity when the female body is in a disease state,and the effects of different stress levels on the female reproductive system and the increase in the risk of female disease will vary greatly.Studies have shown that ER stress is closely related to the occurrence and development of gynecological diseases.Based on the relevant research reports at home and abroad in recent years,this paper summarizes the role of ER stress in several common gynecological diseases to provide new research ideas for preventing,diagnosing,and treating gynecological diseases.

Membranous nephropathy(MN)is the predominant cause of primary nephrotic syn-drome(NS)among adults.The identification of PLA2R as target antigen has brought about a pro-found transformation in the management of MN,offering a basis for the utilization of B-cell deplet-ing agents such as rituximab(RTX).The question of whether early intervention targeting antibodies can effectively impede the progression of MN,contrib-uting to enhanced disease control and long-term renal outcomes for patients,remains further explo-ration.We analyzed demographic data,laboratory parameters,and renal involvement in 13 patients with PLA2R antibody-related MN who received at least one RTX treatment at our center from Octo-ber 2019 to March 2023.Early-stage medium-to-high-risk MN was defined as baseline or admission anti-PLA2R antibody levels exceeding 50 RU/mL,ex-cluding patients who already presented with ne-phrotic syndrome at baseline.The median duration of MN at the initiation of the first RTX treatment was 4.1 months(IQR 1-7.7),and the median follow-up time after RTX therapy was 27 months(IQR 23-45).All patients had commenced renin-angiotensin system inhibitors before receiving RTX.Following RTX therapy,none of the 13 patients progressed to NS during the follow-up period,and 12 patients achieved complete or partial remission at the 2-year follow-up or the last visit.No deaths,severe infections,or other serious adverse reactions oc-curred during the follow-up period.In conclusion,RTX demonstrates favorable efficacy and safety in early-stage,medium-to-high-risk MN patients.Initi-ating antibody clearance therapy in these patients may be beneficial for long-term disease control and distant renal outcomes.

Objective To address the issue of missing entity relationship information in the knowledge graph of TCM ancient books;To propose a knowledge reasoning method based on deep reinforcement learning;To improve the completeness of the knowledge graph.Methods Based on the benchmark dataset in the field of knowledge graph and the dataset of TCM ancient books,a deep reinforcement learning inference method was adopted.Combined with knowledge one-step completion and multi hop path search,Python language was used in conjunction with the Neo4j graph database to achieve path to path inference.Results In the knowledge reasoning task dataset,compared with the baseline model,this reasoning method improved the MAP index by about 53.79%.When applied to the knowledge graph of TCM ancient books,its evaluation index reached the highest of 0.776.Conclusion The knowledge reasoning method based on deep reinforcement learning has significant advantages in improving the completeness of the knowledge graph of TCM ancient books,and this method can effectively fill the information gap in the graph of TCM ancient books.

Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.