BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

Objective To clarify the risk factors of diaphragmatic dysfunction (DD) after cardiac surgery with extracorporeal circulation. Methods A retrospective analysis was conducted on the data of patients who underwent cardiac surgery with extracorporeal circulation in the Department of Cardiovascular Surgery of Peking University People's Hospital from January 2023 to March 2024. Patients were divided into two groups according to the results of bedside diaphragm ultrasound: a DD group and a control group. The preoperative, intraoperative, and postoperative indicators of the patients were compared and analyzed, and independent risk factors for DD were screened using multivariate logistic regression analysis. Results A total of 281 patients were included, with 32 patients in the DD group, including 23 males and 9 females, with an average age of (64.0±13.5) years. There were 249 patients in the control group, including 189 males and 60 females, with an average age of (58.0±11.2) years. The body mass index of the DD group was lower than that of the control group [(18.4±1.5) kg/m2 vs. (21.9±1.8) kg/m2, P=0.004], and the prevalence of hypertension, chronic obstructive pulmonary disease, heart failure, and renal insufficiency was higher in the DD group (P<0.05). There was no statistical difference in intraoperative indicators (operation method, extracorporeal circulation time, aortic clamping time, and intraoperative nasopharyngeal temperature) between the two groups (P>0.05). In terms of postoperative aspects, the peak postoperative blood glucose in the DD group was significantly higher than that in the control group (P=0.001), and the proportion of patients requiring continuous renal replacement therapy was significantly higher than that in the control group (P=0.001). The postoperative reintubation rate, tracheotomy rate, mechanical ventilation time, and intensive care unit stay time in the DD group were higher or longer than those in the control group (P<0.05). Multivariate logistic regression analysis showed that low body mass index [OR=0.72, 95%CI (0.41, 0.88), P=0.011], preoperative dialysis [OR=2.51, 95%CI (1.89, 4.14), P=0.027], low left ventricular ejection fraction [OR=0.88, 95%CI (0.71, 0.93), P=0.046], and postoperative hyperglycemia [OR=3.27, 95%CI (2.58, 5.32), P=0.009] were independent risk factors for DD. Conclusion The incidence of DD is relatively high after cardiac surgery, and low body mass index, preoperative renal insufficiency requiring dialysis, low left ventricular ejection fraction, and postoperative hyperglycemia are risk factors for DD.

Attention is the cornerstone of effective functioning in a complex and information-rich world. While the neural activity of attention has been extensively studied in the cortex, the brain-wide neural activity patterns are largely unknown. In this study, we conducted a comprehensive analysis of neural activity across the mouse brain during attentional processing using EEG and c-Fos staining, utilizing hierarchical clustering and c-Fos-based functional network analysis to evaluate the c-Fos activation patterns. Our findings reveal that a wide range of brain regions are activated, notably in the high-order cortex, thalamus, and brain stem regions involved in advanced cognition and arousal regulation, with the central lateral nucleus of the thalamus as a strong hub, suggesting the crucial role of the thalamus in attention control. These results provide valuable insights into the neural network mechanisms underlying attention, offering a foundation for formulating functional hypotheses and conducting circuit-level testing.
Animals ; Attention/physiology* ; Mice ; Brain/physiology* ; Male ; Electroencephalography ; Reaction Time/physiology* ; Brain Mapping ; Mice, Inbred C57BL ; Choice Behavior/physiology* ; Proto-Oncogene Proteins c-fos/metabolism*

This study investigated the epidemic status of H3N2 influenza virus and the genetic evolution characteristics of hemagglutinin(HA)and neuraminidase(NA)of H3N2 subtype influenza viruses isolated in Shandong Province during 2023-2024,to understand their compatibility with vaccine strains and drug resistance status.A total of 25 H3N2 subtype influenza virus strains were randomly selected from the strains isolated by the influenza surveillance network laboratory.The HA and NA genes were sequenced with the vaccine strains recommended by the WHO as a reference.Monitoring of sensitivity to oseltamivir and zanamivir was conducted through neuraminidase inhibition experiments.The H3N2 influenza viruses in Shandong Province belonged to the 3C.2a1b.2a.2a.3a.1 clade.Nucleotide sequence analysis revealed that the HA1 and NA genes were closely related to the WHO-recommended vaccine strain A/Darwin/9/2021 for the current season,with homology rates of 97.8%-98.2%and 98.9%-99.3%,respectively.Amino acid sequence analysis indicated 22 amino acid sequence variations in the HA1 protein,and antigenic drift was detected in 8 strains.A glycosylation site was added at position 94 of the HA protein in all 25 strains.Variations occurred in the NA antigenic determinants of some strains.Neuraminidase inhibition experiments indicated that all tested influenza viruses were sensitive to oseltamivir and zanamivir.Some differences in HA and NA proteins were observed between the monitored strains and vaccine strains.Continued monitoring of the molecular evolution characteristics of influenza viruses is necessary to understand the risk of influenza outbreaks,and their effects on the effectiveness of influenza vaccines and therapeutic drugs.

Objective:To analyze the epidemiological, etiological and genetic characteristics of influenza virus in Shandong Province during 2023-2024.Methods:The surveillance data of influenza-like illness (ILI) in sentinel hospitals in Shandong from 2023 to 2024 were collected and analyzed. The isolated influenza strains with hemagglutination titers ≥8 were selected for antigenicity analysis, drug susceptibility test, gene sequencing and evolutionary analysis.Results:From 2023 to 2024, the positive rate of influenza virus in Shandong was 8.51% (23 663/277 995), the highest positive rate was in the age group of 5-14 years (15.78%, 6 073/38 478), and the highest positive rate was in the 49 th week (35.86%, 2 264/6 313). Both antigenicity analysis and evolutionary analysis showed that the A(H1N1)pdm09 subtype and B(Victoria) strain had good matching effect and close evolutionary distance with the 2023-2024 surveillance year vaccine strain. The A(H3N2) subtype strain did not have a high matching effect with the 2023-2024 vaccine strain and had a long evolutionary distance, but had a close evolutionary distance with the 2024-2025 vaccine strain. Drug susceptibility test showed that oseltamivir sensitivity of influenza A(H1N1)pdm09 strain decreased greatly, and the amino acid site mutation of neuraminidase was H275Y. Conclusions:In the 2023-2024 surveillance year, the peak of influenza virus epidemic in Shandong was mainly occurred in winter and spring, and the age group of 5-14 years was the focus of prevention and control. The dominant strain was subtype A(H3N2), which had poor matching effect with the vaccine strain in the 2023-2024 surveillance year. One A(H1N1)pdm09 resistant strain was found in the drug resistance monitoring work. Follow-up prevention and control work should be strengthen the surveillance for the epidemiological characteristics, genetic variation and drug resistance of influenza viruses, timely understand the epidemic trend and mutation of influenza viruses, timely discover drug-resistant strains of influenza viruses, promote influenza vaccination, and improve of influenza prevention and control.

Objective:To deeply analyze the epidemiological characteristics and changing trends of intestinal infectious diseases in China, and provide scientific evidence for the prevention and control of intestinal infectious diseases.Methods:The incidence data of notifiable intestinal infectious diseases in China from 2013 to 2022 were collected from China Disease Prevention and Control Information System. Descriptive statistical method was used to analyze the distributions of intestinal infectious diseases in China, and the annual change rate and seasonal index were calculated.Results:During 2013-2022, intestinal infectious diseases were reported nationwide, with the cases accounting for 43.50% of all notifiable infectious disease cases. The average reported incidence rate was 224.50/100 000, showing a decreasing trend year by year (average annual percent change=-6.45%, t=-2.76, P=0.025). The top 5 intestinal infectious diseases were hand foot and mouth disease (HFMD) (130.40/100 000), other infectious diarrhea (80.18/100 000), dysentery (7.45/100 000), acute hemorrhagic conjunctivitis (2.49/100 000) and viral hepatitis E (1.92/100 000). The incidences of dysentery, HFMD, typhoid fever/paratyphoid fever, viral hepatitis A and acute hemorrhagic conjunctivitis all showed decreasing trends year by year (all P<0.05), while the incidences of hepatitis E and other infectious diarrhea showed no significant changes with year (both P>0.05). The incidence of intestinal infectious diseases was high during May to October, with the peak in June. The incidence rate of intestinal infectious diseases was significantly higher in men than in women (all P<0.05). The HFMD, other infectious diarrhea and dysentery cases were mainly children aged 0-5 years, while the cholera, hepatitis A, hepatitis E, typhoid fever/paratyphoid fever and acute hemorrhagic conjunctivitis cases were mainly farmers aged ≥20 years. The annual reported incidence rate of intestinal infectious diseases was higher in southern provinces (283.66/100 000) than in northern provinces (142.63/100 000), and the annual reported incidence rate of intestinal infectious diseases was higher in coastal provinces (279.52/100 000) than in inland provinces (181.78/100 000), the differences were all significant (both P<0.001). Conclusions:During 2013-2022, the incidence of intestinal infectious diseases decreased significantly in China, with HFMD and other infectious diarrhea as the main diseases. Strengthened surveillance for intestinal infectious diseases should be carried out in key groups, such as children living scatteredly and farmers, and targeted prevention and control measures should be taken according to the epidemiological characteristics of different diseases to effectively reduce the incidence of intestinal infectious diseases.

Objective:To analyze the situation of rodents carrying Hantavirus and the genetic and evolutionary characteristics of the virus in Zibo city, Shandong province in 2022, and provide reference for the scientific prevention and control of hemorrhagic fever with renal syndrome (HFRS).Methods:Real-time quantitative PCR (RT-qPCR) was used to detect hantavirus (HV) nucleic acids in rodent lung tissues and identify HV genotypes. Each nucleic acid fragment was designed to amplify various gene fragments by segment, and the whole genome of Hantavirus was sequenced by second generation sequencing. Sequence assembly was performed using SeqMan 7.1.0.44, a subprogram of DNAStar. Sequence alignment and evolutionary analysis were conducted using MEGA 7.0 and BioEdit software.Results:A total of 270 host animals were captured in this survey. Among them, 13 rodent lung samples tested positive for Hantavirus, resulting in a virus-positive rate of 4.8%. The full-genome sequences of four hantavirus strains were successfully obtained, all identified as Seoul virus (SEOV) genotype. Four Hantavirus-positive samples showed high nucleotide sequence homology in the M gene and belonged to the SEOV S3 subtype. These strains exhibited high similarity with those from Hebei, Liaoning, and Beijing. The amino acid sequences of the nucleoprotein and glycoprotein immunogenic epitopes were identical to those of the vaccine strain Z37.Conclusions:This study successfully determined the full genome sequences of four hantavirus strains from Zibo city, Shandong province. The genotypes are primarily SEOV, with the subtype being S3. The homology of genes within the same subtype is high, with no significant variations observed. The alignment of immune epitopes in key proteins suggests that the current vaccine may provide protection against locally circulating strains, but further in-depth research is still required.

Objective:To understand the epidemiological characteristics of severe fever with thrombocytopenia syndrome (SFTS) in Shandong province from 2010 to 2022, analyze the trends in morbidity and mortality, and provide a scientific basis for the prevention, control, diagnosis, and treatment of SFTS.Methods:The data on SFTS cases and deaths were retrospectively collected through the China Disease Control and Prevention Information System′s infectious disease surveillance system and the Shandong province all-cause mortality surveillance system. Descriptive epidemiological method was used for epidemiological analysis, and the Joinpoint regression model was applied to analyze trends in morbidity and mortality.Results:From 2010 to 2022, 6 714 SFTS cases were reported in Shandong, with an average crude incidence rate (CIR) and age-standardized incidence rate (ASIR) of 0.52/100 000 and 0.43/100 000, respectively. Among these, 1, 064 cases died within 30 days of onset, with an average crude mortality rate (CMR) and age-standardized mortality rate (ASMR) of 0.08/100, 000 and 0.06/100 000, respectively, and an average case fatality rate of 15.85%. The average incidence rates for males and females were 0.52/100 000 and 0.53/100 000, respectively, with no statistically significant difference ( P>0.05). The average annual mortality rate for males (0.09/100 000) was higher than that for females (0.08/100 000), and the average annual case fatality rate for males (17.21%) was higher than that for females (14.48%), with statistically significant differences ( P<0.05); 93.95% of the cases were over 45 years old, and 86.56% of the deaths were over 60 years old. Both cases and deaths were primarily among farmers, accounting for over 85% of the total. From 2010 to 2022, the incidence rate of SFTS in Shandong showed an overall upward trend, with an average annual percentage change (AAPC) in CIR of 23.58% and an AAPC in ASIR of 19.97%, P<0.05. The mortality rate of SFTS cases in Shandong from 2010 to 2022 showed a continuous and significant upward trend, with a CMR AAPC of 27.64% and an ASMR AAPC of 22.70%, P<0.05. SFTS incidence and mortality in Shandong exhibited clear seasonality, with high occurrence from May to October, primarily concentrated in the Jiaodong Peninsula and the hilly areas of central Shandong. The number of affected counties increased in a wave-like pattern, expanding from 6 in 2010 to 79 in 2021 for cases and from 2 in 2010 to 53 in 2022 for deaths. Conclusions:From 2010 to 2022, the incidence and mortality of SFTS in Shandong showed an upward and expanding trend, with a particularly heavy disease burden among middle-aged and elderly farmers.