ObjectiveTo analyze the epidemiological trends and characteristics of brucellosis in humans (hereinafter referred to as brucellosis) in Tangshan City, Hebei Province from 2016 to 2023, and to provide a scientific basis for formulating brucellosis prevention and control strategies in the region. MethodsThe incidence data of human brucellosis in Tangshan City from 2016 to 2023 were collected from the China Disease Prevention and Control Information System. The diagnosis time, infection route, and clinical characteristics of the cases were obtained from the case investigation reports. Descriptive epidemiological methods were used to analyze the temporal, spatial, demographic distributions, and clinical characteristics of human brucellosis. Brucella species were identified using agglutination tests with bacterial suspension and A/M antigen-positive serum. ResultsA total of 2 193 cases of human brucellosis were confirmed and clinically diagnosed in Tangshan City from 2016 to 2023, with the peak incidence occured from March to August, and which exhibited distinct geographic distribution patterns. The highest incidence rate was found in people aged 60‒<70 years. The occupation of cases were primarily farmers. The incidence rate in males (528/100 000) was higher than that in females (184/100 000). All cases had confirmed exposure to infected animals or contaminated animal products. ConclusionThe epidemic of human brucellosis in Tangshan exhibited an overall steady downward trend from 2016 to 2023, except for a slight increase in 2016 and 2021, with the incidence rate controlled at 289/100 000‒335/100 000. The prevention and control situation of human brucellosis still remains severe, with the highest incidence rate in the eastern region of Tangshan, which are characterized by the breeding, slaughtering, and processing of cattle and sheep. Therefore, it it is necessary to enhance the prevention and control of human brucellosis among the personnel engaged in these industries in the eastern areas.

T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive hematologic malignancy with a poor prognosis, despite advancements in treatment. Many patients struggle with relapse or refractory disease. Investigating the role of the super-enhancer (SE) regulated gene ubiquitin-specific protease 20 (USP20) in T-ALL could enhance targeted therapies and improve clinical outcomes. Analysis of histone H3 lysine 27 acetylation (H3K27ac) chromatin immunoprecipitation sequencing (ChIP-seq) data from six T-ALL cell lines and seven pediatric samples identified USP20 as an SE-regulated driver gene. Utilizing the Cancer Cell Line Encyclopedia (CCLE) and BloodSpot databases, it was found that USP20 is specifically highly expressed in T-ALL. Knocking down USP20 with short hairpin RNA (shRNA) increased apoptosis and inhibited proliferation in T-ALL cells. In vivo studies showed that USP20 knockdown reduced tumor growth and improved survival. The USP20 inhibitor GSK2643943A demonstrated similar anti-tumor effects. Mass spectrometry, RNA-Seq, and immunoprecipitation revealed that USP20 interacted with hypoxia-inducible factor 1 subunit alpha (HIF1A) and stabilized it by deubiquitination. Cleavage under targets and tagmentation (CUT&Tag) results indicated that USP20 co-localized with HIF1A, jointly modulating target genes in T-ALL. This study identifies USP20 as a therapeutic target in T-ALL and suggests GSK2643943A as a potential treatment strategy.

Objective:To investigate the potential mechanism of miR-1298-5p in non-small cell lung cancer(NSCLC)to regu-late the MSH2 gene and its effect on the biological behavior of tumor cells and the tumor immune microenvironment.Methods:Bioin-formatics was used to identify the key genes and miRNA involved in NSCLC.Cell proliferation was detected by CCK-8 assay,and cell invasion and migration were detected by Transwell assay.Levels of inflammatory factors were detected by ELISA assay.Western blot was used to measure the expression of MSH2 in cells,and fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expressions of miR-1298-5p and MSH2 gene in NSCLC cells.Dual luciferase reporter gene assay was performed to verify the targeting relationship between miR-1298-5p and MSH2.Spearman correlation analysis was performed to correlate miR-1298-5p with immune cells and immune factors in the tumor immune microenvironment.Results:The level of miR-1298-5p was down-regulated in NSCLC cells compared with normal lung tissue cells.miR-1298-5p overexpression inhibited the proliferation,migration and inva-sion of NSCLC cells.MSH2 was confirmed to be a target gene of miR-1298-5p using luciferase reporter gene assay.Furthermore,down-regulation of miR-1298-5p in NSCLC cells could be reversed by silencing MSH2.miR-1298-5p expression levels were negatively corre-lated with the levels of Treg,IL-10,and TGF-β,and positively correlated with the levels of CD3+T,CD4+T,CD8+T,NK cells,IL-2,and IFN-γ.Conclusion:miR-1298-5p negatively regulates MSH2 to inhibit the proliferation,invasion and migration of NSCLC cells and improve the tumor immune microenvironment.

Objective:To explore the application of PDCA (plan, do, check, act) cycle theory in the teaching of eight-year clinical medical students for the course of Laboratory Diagnostics.Methods:From September 2021 to November 2022, the students majoring in clinical medicine of eight-year system who were studying the course of Laboratory Diagnostics at Peking Union Medical College were selected as the research objects. The teaching reform of the course of Laboratory Diagnostics was carried out according to the four stages of PDCA cycle theory, and the teaching effect was evaluated by the final examination results and questionnaire survey scores of the students.Results:The score of theoretical examination of eight-year students in 2018 was (86.7±4.68) points, which was higher than that of students in 2017 [(83.3±3.89) points], and the difference was statistically significant ( P<0.05). The questionnaire survey results of the two groups of students showed that the rating of experimental course was higher than that of theoretical course ( P<0.05). After the teaching reform based on PDCA cycle theory, the questionnaire survey scores of theoretical course [(3.83±0.25) points vs (2.94±0.28) points] and experimental course [(4.13±0.09) points vs (3.32±0.12) points] in students of 2018 were higher than those of 2017 (all P<0.001). Conclusion:PDCA cycle theory provides new methods and ideas for teaching management, which helps to improve the performance of clinical medical students of eight-year system and their recognition of the course.

Based on Donabedian three-dimensional quality structure theory, this paper summarized the sensitive quality indicators of breast cancer nursing and summarized the research status from three aspects of structure, process and quality of results, to analyze the shortcomings of current research, and propose improvement strategies. The aim is to provide reference for the construction of scientific, standardized, practical and specialized sensitive quality indicators of breast cancer nursing in accordance with China′s national conditions.

Objective:To explore the value of iterative decomposition of water and fat with asymmetry and least squares estimation-quantitative fat imaging (IDEAL-IQ) in quantitative evaluation of thigh muscle fat content and its correlation with muscle strength in middle-aged and elderly volunteers.Methods:From December 2020 to April 2021, 30 volunteers aged 45 to 70 were recruited prospectively, including 15 males and 15 females with 52.5 (49.0, 56.3) years old. All subjects were scanned at 3.0 T MR, including axial T 1WI, IDEAL-IQ and coronal T 2WI of the left thigh. The region of interest of the knee extensors (quadriceps femoris) and knee flexors (hamstrings) in the left mid-thigh were delineated, and muscle cross-sectional area (CSA), skeletal muscle index (SMI), intermuscular fat fraction (FF) and intramuscular FF were obtained. In addition, isokinetic muscle strength measurement was performed on the left knee joint of all subjects at angular speeds of 60°/s and 180°/s to obtain peak torque (PT) and total work (TW) of knee flexors and extensors. Independent sample t-test, paired t-test or Mann-Whitney U test were used to compare the differences of CSA, SMI, intermuscular FF, intramuscular FF, PT and TW between different genders and muscle groups. Pearson or Spearman correlation analysis, and multiple linear regression analysis were used to analyze the correlation between CSA, SMI, intermuscular FF, intramuscular FF and PT, TW of thigh muscles. Results:The CSA, PT and TW of thighs in males were higher than those in females ( P<0.05), while the intermuscular FF in males was lower than that in females ( P=0.005). The CSA, SMI and PT of the thigh extensors were higher than those of the flexors ( P<0.001), while the intramuscular FF and intermuscular FF were lower than those of the flexors ( P<0.001). Intramuscular FF of flexors and extensors were moderately negatively correlated with PT ( r=-0.635, P<0.001; r=-0.546, P<0.001), and highly, moderately negatively correlated with TW ( r=-0.718, P<0.001; r=-0.616, P<0.001). Intermuscular FF of flexors and extensors were moderately negatively correlated with PT ( r=-0.519, P=0.003; r=-0.443, P=0.014), and negatively correlated with TW ( r=-0.363, P=0.049; r=-0.552, P=0.002). There was no significant correlation between CSA, SMI and PT, TW in flexors and extensors of thigh ( P>0.05). Multiple linear regression analysis showed that intramuscular FF was still significantly correlated with PT and TW of flexors and extensors (flexors: R 2adj=0.505, P=0.001; R 2adj=0.540, P<0.001; extensors: R 2adj=0.351, P=0.006; R 2adj=0.470, P=0.002). Conclusion:FF based on IDEAL-IQ technology can accurately quantify the intramuscular and intermuscular fat content of thighs, and there are negative correlations between intramuscular FF, intermuscular FF and isokinetic muscle strength measurements including PT and TW. Among them, intramuscular FF is more significant.

Objective:To investigate the effect of heme oxygenase-1 (HO-1) modified bone marrow mesenchymal stem cells (BMMSCs) combined with normothermic machine perfusion (NMP) on the intestinal barrier function in rats with acute rejection of liver transplantation.Methods:Specific pathogen free 2 male Brown Norway (BN) rats (4-5 weeks, 40-60 g) were used to isolat BMMSCs, and HO-1 was infected by adenovirus. Of 24 male Lewis rats (7-8 weeks old, 200-220g) were used as donors, 30 male BN rats (8-9 weeks old, 220-240 g) were used as recipients. Acute rejection models of orthotopic liver transplantation were established in rats using two cuff technique. BN recipient rats were randomly divided into five groups: sham group, abdomen of the mice was open and closed within 30 min; NMP livers were simply mechanically perfused for 4 h; the BMP group were perfused with BMMSCs through the portal vein; the HBP group were perfused with HO-1/BMMSCs through the portal vein; the FK506 livers were mechanically perfused for 4 h and administered intragastrically of tacrolimus daily following surgery, 6 per group, on days 14 after surgery, the relevant indicators were taken and the rejection activity index (RAI) changes were investigated. The changes of intestinal pathological were analyzed by HE staining and transmission electron microscope, the expression levels of zonula occludens-1 (ZO-1) and occludin protein in intestinal tissue were detected by Western blotting, the concentrations of lipopolysaccharide, D-lactic acid and diamine oxidase (DAO) in serum were detected by ELISA.Results:The RAI of HBP group (2.80±0.84) and FK506 group (2.20±0.84) were significantly lower than that of NMP group (7.60±1.14) and BMP group (6.00±1.58), the differences were statistically significant (all P<0.05). The intestinal villi in NMP group were significantly sparse, wrinkled and disorderly arranged while the degree of intestinal injury in BMP group, HBP group and FK506 group were more mitigated. Electron microscope observation showed that the microvilli of intestinal epithelial cells in HBP group were rich and orderly, and the tight junction structure between cells was complete. The protein expression levels of ZO-1 and Occludin in the intestinal tissues of HBP group [(0.87±0.06) (1.28±0.26)] were higher than those of NMP group [(0.41±0.12) (0.27±0.18)] and FK506 group [(0.52±0.15) (0.63±0.22)], the differences were statistically significant (all P<0.05). The concentration of lipopolysaccharide, D-lactic acid and DAO in serum of HBP group was lower than those of NMP group and FK506 group, the differences were statistically significant (all P<0.05). Conclusion:HO-1/BMMSCs combined with NMP protects the intestinal mucosal barrier function of BN rats with acute rejection after liver transplantation.

Objective:To explore the effect and possible mechanism of normal temperature mechanical perfusion(NMP)plus bone marrow mesenchymal stem cells(BMMSCs)on early endoplasmic reticulum stress(ERs)and cellular apoptosis after donation after cardiac death(DCD)donor liver transplantation.Methods:BMMSCs were isolated and cultured by adherence method.Sixty Sprague-Dawley(SD)rats were randomly divided into five groups of sham operation(sham), static cold storage(SCS), NMP, BMMSC and NMP plus BMMSCs(BP)( n=12 each). Liver tissue and serum sample of each group were harvested at Day 1/7 post-operation.Hematoxylin-eosin(HE)staining was employed for observing pathological changes of liver tissue; TdT-mediated dUTP nick end labeling(TUNNEL)staining for detecting cellular apoptosis; immunohistochemistry for detecting the expression of hepatocyte transcription factor C/EBP homologous protein(CHOP); Western blot for detecting the expressions of GRP-78, p-PERK, ATF4, CHOP and cleaved caspase-3. Results:Compared with SCS group, hepatic injury and inflammation significantly declined in NMP, BMMSC and BP groups and improvement of hepatic injury was the most pronounced in BP group.Cellular apoptosis lessened markedly in BP group at Day 1/7 as compared with SCS group and the difference was statistically significant.The expressions of ERs-related proteins GRP-78, p-PERK and ATF4 spiked in SCS group and the expressions of pro-apoptotic proteins CHOP and cleaved caspase-3 were significantly elevated and declined markedly in BP group.And the difference was statistically significant.Conclusions:BMMSCs plus NMP can significantly improve hepatocyte apoptosis and inflammatory response after DCD donor liver transplantation.And its mechanism may be correlated with suppressing early endoplasmic reticulum stress of hepatocytes.

Objective To evaluate the effect of 68 Ga-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-D-Phe1-Tyr3-Thr8-octreotide ( DOTATATE) PET/CT imaging combined with MRI on im-proving the diagnostic efficiency for tumor-induced osteomalacia (TIO). Methods From March 2012 to October 2018, 159 patients (89 males, 70 females; age 17-59 years) who were suspected with TIO and underwent both 99 Tcm-hydrazinonicotinamide-Tyr3-octreotide ( HYNIC-TOC ) SPECT imaging and 68 Ga-DOTATATE PET/CT imaging were recruited into this retrospective study. Lesions presenting abnormal in-creased focal uptake in any part of the body were suspected of culprit tumors, excluding those induced by fractures and inflammation. Based on the suspicious culprit lesions detected by 68 Ga-DOTATATE PET/CT imaging, 80 patients underwent corresponding MRI. The lesion presenting as the space-occupying nodule or mass with abnormal signal on MR image was diagnosed as the TIO culprit lesion. Compared with the results of pathological examination and clinical follow-up, the diagnostic efficiency of 99 Tcm-HYNIC-TOC SPECT imaging and 68 Ga-DOTATATE PET/CT imaging and the combination of 68 Ga-DOTATATE PET/CT imaging and MRI for TIO were analyzed. Results Of 159 patients, 135 were confirmed with TIO and 24 were with non-TIO. 99 Tcm-HYNIC-TOC SPECT imaging accurately diagnosed 92 TIO patients and 22 non-TIO pa-tients, with the sensitivity, specificity and accuracy of 68.1％(92/135), 91.7％(22/24) and 71.7％(114/159) respectively. 68 Ga-DOTATATE PET/CT imaging accurately diagnosed 134 TIO patients and 19 non-TIO patients, with the sensitivity, specificity and accuracy of 99.3％(134/135), 79.2％(19/24) and 96. 2％(153/159) respectively. A total of 74 TIO and 5 non-TIO patients were accurately diagnosed by 68Ga-DOTATATE PET/CT imaging combined with MRI, with the sensitivity, specificity and accuracy of 98.7％(74/75), 5/5 and 98.8％(79/80). Conclusion 68Ga-DOTATATE PET/CT imaging shows higher sensi-tivity than 99 Tcm-HYNIC-TOC SPECT imaging in diagnosing TIO, and the combination of 68 Ga-DOTATATE PET/CT imaging and MRI can improve the specificity and accuracy.

To evaluate the immunogenicity of HA globular head domain of H5 subtype influenza virus (H5HA), the gene of H5HA was optimized and the recombinant pPICZaA-H5HA expressing vector was constructed and transfected into Pichia pastoris. The expression of the recombinant H5HA was confirmed by SDS-PAGE and Western blotting and the results demonstrated that the recombinant H5HA (37 kDa) was highly expressed in Pichia pastoris with concentration of 0.2 mg/mL in medium. The recombinant H5HA was concentrated and purified using Ni-NTA affinity chromatography. The immunogenicity of H5HA was evaluated by immunizing eight groups of chicken through intranasal or intramuscular injection with different doses of purified H5HA combined with different adjuvants, respectively. The results showed that the recombinant H5HA could induce high level IgG (HI titer was 1:64 and neutralizing antibody titer was 1:218) and the optimal dosage of the recombinant H5HA was 50 μg combined with oil. In addition, intramuscular injection was better than nasal immunization. This study provided a theoretical support for subunit vaccine development.
Animals ; Antibodies, Viral ; Birds ; Hemagglutinin Glycoproteins, Influenza Virus ; Influenza A Virus, H5N1 Subtype ; Influenza Vaccines ; Influenza in Birds ; Pichia ; Vaccination