Objective To research the microbial population distribution on the outer surface and in the contents of SPF chicken embryos, so as to standardize the production of SPF chicken embryos, guide the selection of disinfectants for vaccine production, and improve the sterile guarantee level of vaccine production.Methods The cotton swab method was used to isolate the external surface microorganisms of three batches of SPF chicken embryos before and after treatment with 70%isopropanol. The microorganisms of SPF chicken embryo contents were isolated with tryptose soya agar(TSA) medium from the urine sac fluid of three batches of SPF chicken embryos that died after 3 d inoculation with attenuated yellow fever 17D strain. The colony was purified using the three-zone streaking method, and the microorganisms were identified using 16s rRNA/ITS sequencing.Results After disinfection with 70% isopropanol, the number of microorganisms detected on the outer surface of SPF chicken embryos significantly decreased(t = 2. 67, P = 0. 011). The microorganisms on the outer surface of SPF chicken embryos were mainly Bacillus, with Bacillus genus accounting for 52. 6% and other types of bacillus accounting for36. 8%. There were significant differences in the detection rates of microorganisms in the contents of three batches of SPF chicken embryos. Proteobacteria, Enterococcus and Escherichia were detected the most frequently in the contents of SPF chicken embryos, and their detection numbers were significantly higher than other microorganisms.Conclusion The dominant microorganisms on the outer surface of SPF chicken embryos are Bacillus, and the dominant microorganisms in the contents are Proteobacteria, Enterococcus and Escherichia. The microbial population distribution provides basis for the selection of disinfectants for vaccine production, suggesting that the control measures should be carried out at multiple stages such as SPF chicken embryos production, storage, transportation, and selection to improve the sterile assurance level of vaccine production.

Objective To quantitatively assess the association of short-term exposure to polycyclic aromatic hydrocarbons (PAHs) in ambient fine particulate matter (PM2.5) with residents mortality. Methods A time-stratified case-crossover study was conducted from 2020 to 2022 among 10606 non-accidental residents by using the Guangzhou Cause of Death Surveillance System in Conghua District, Guangzhou. Exposure levels of PAHs in PM_2.5 and meteorological data during the study period were obtained from the Center for Disease Control and Prevention in Conghua District and the China Meteorological Administration Land Data Assimilation System (CLDAS-V2.0), respectively. Conditional Poisson regression model was used to estimate the exposure-response association between PAHs and the mortality risk. Results Fluoranthene, chrysene, benzo[k]fluoranthene, benzo[a]pyrene, and indeno[1,2,3-cd]pyrene were significantly associated with an increased risk of mortality. For every one interquartile range increase in exposure levels, the non-accidental mortality risks increased by 8.33% (95% CI: 1.80%, 15.27%), 4.67% (95% CI: 1.86%, 7.57%), 6.07% (95% CI: 2.08%, 10.21%), 4.62% (95% CI: 1.85%, 7.47%), and 4.70% (95% CI: 0.53%, 9.03%), respectively. The estimated non accidental deaths attributable to exposure to fluoranthene, chrysene, benzo[k]fluorine, benzo[a]pyrene and indine[1,2,3-cd]pyrene were 5.91%, 6.08%, 6.51%, 6.46%, and 4.21%, respectively. Conclusions Short-term exposure to PAHs in PM_2.5, including fluoranthene, chrysene, benzo[k]fluoranthene, benzo[a]pyrene and indine[1,2,3-cd]pyrene, was significantly associated with an increased risk of mortality among residents.

Acute lung injury (ALI) is a severe disease caused by viral infection that triggers an uncontrolled inflammatory response. This study investigated the capacity of jasurolignoside (JO), a natural compound, to bind to Toll-like receptor 4 (TLR4) and treat ALI. The anti-inflammatory properties of JO were evaluated in vitro through Western blotting, enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining, and co-immunoprecipitation. The investigation utilized a lipopolysaccharide (LPS)-induced ALI animal model to examine the therapeutic efficacy and mechanism of JO in vivo. JO attenuated inflammatory symptoms in infected cells and tissues by modulating the NOD-like receptor family pyrin domain containing protein 3 (NLRP3) inflammasome and the nuclear factor κB (NF-κB)/mitogen-activated protein kinase (MAPK) pathway. Molecular docking simulations revealed JO binding to TLR4 active sites, confirmed by cellular thermal shift assay. Surface plasmon resonance (SPR) demonstrated direct interaction between JO and TLR4 with a Kd value of 35.1 μmol·L^-1. Moreover, JO inhibited tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and IL-6 secretion and reduced leukocyte, neutrophil, lymphocyte, and macrophage infiltration in ALI-affected mice. JO also enhanced lung function and reduced ALI-related mortality. Immunohistochemical staining demonstrated JO's ability to suppress TLR4 expression in ALI-affected mouse lung tissue. This study establishes that JO can bind to TLR4 and effectively treat ALI, indicating its potential as a therapeutic agent for clinical applications.
Toll-Like Receptor 4/chemistry* ; Animals ; Acute Lung Injury/chemically induced* ; Mice ; Humans ; Ilex/chemistry* ; Molecular Docking Simulation ; Male ; NF-kappa B/immunology* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology* ; Tumor Necrosis Factor-alpha/genetics* ; Interleukin-1beta/genetics* ; RAW 264.7 Cells ; Disease Models, Animal

Background Exposure to air pollutants increases the risk of diseases in multiple systems, including respiratory and cardiovascular systems, yet its association with neurological diseases remains unclear. Objective To quantitatively evaluate the association between short-term exposure to air pollutants and outpatient and emergency visits for neurological diseases, identify potential susceptible populations, and quantify associated disease burden. Methods Daily 24-hour average concentrations of fine particulate matter (PM_2.5), inhalable particulate matter (PM₁₀), sulfur dioxide (SO₂), nitrogen dioxide (NO₂), and carbon monoxide (CO), daily maximum 8-hour average concentration of ozone (O₃), daily meteorological data (24-hour average temperature, 24-hour average relative humidity), and data on daily outpatient and emergency department visits for neurological diseases from two hospitals in Conghua District, Guangzhou, China, were collected from 2015 to 2022. A time-stratified case-crossover design was adopted, and a conditional Poisson regression model was constructed to analyze the association between air pollution exposure and neurological disease visits. Two-pollutant models and sensitivity analysis were used to validate model stability. Stratified analyses by season (cold season: from November to March; warm season: from April to October), sex (male, female), and age (≤45 years, 46–60 years, ≥61 years) were performed to identify vulnerable group. Additionally, the number and proportion of neurological disease visits attributable to short-term air pollutant exposure were calculated. Results A total of 72 673 outpatient and emergency department visits for neurological diseases were included during the study period. Most of the patients were middle-aged and elderly individuals (69.89% were over 45 years old) and females (60.25%). The results of single-pollutant models showed that for each interquartile range (IQR) increase in exposure to PM_2.5, PM₁₀, SO₂, NO₂, CO, and O₃, the risk of outpatient and emergency department visits for neurological diseases increased by 7.54% (95%CI: 4.69%, 10.46%), 6.66% (95%CI: 3.92%, 9.46%), 16.72% (95%CI: 10.58%, 23.19%), 8.12% (95%CI: 4.82%, 11.53%), 5.60% (95%CI: 2.34%, 8.97%), and 6.11% (95%CI: 2.91%, 9.40%), respectively. The results of the two-pollutant model showed that the association between PM_2.5 and SO₂ exposure and outpatient and emergency department visits for neurological diseases were relatively stable. The stratified analyses showed that the effect of SO₂ was stronger in the cold season. It was estimated that 8.32% (95%CI: 5.55%, 10.96%) and 6.65% (95%CI: 4.27%, 8.96%) of the outpatient and emergency department visits were attributable to short-term exposure to SO₂ and PM_2.5, respectively. Conclusion Exposure to PM_2.5 and SO₂ is associated with increased risks of outpatient and emergency visits for neurological diseases. SO₂ shows stronger effects during the cold season, and exposure to air pollution contributes to up to 8.32% of neurological disease visits.

Objective To analyze the current research status of fibrobronchial bronchoalveolar lavage(BAL)in China,explore the development context and research hotspots.Methods This study was conducted from 2003 to 2023 by searching the Chinese literatures on fibrobronchial BAL collected from China National Knowledge Infrastructure,Wanfang Data Knowledge Service Platform,VIP and SinoMed,and extracting the information of keywords,authors,institutions and publication time of the literatures.Co-Occurrencel4.9,VOSviewer 1.6.19 and Microsoft Excel 2021 were applied to carry out data cleaning,data statistics and graph rendering,and visualized the research hotspots and trends of fibrobronchial BAL.Results A total of 4219 literatures meeting the requirements were included.The largest number of articles will be 438 in 2021.There were 522 core authors with more than 3 published papers.The institutions and journals with the largest number of publications were Hebei Children's Hospital with 26 articles and Journal of Clinical Pulmonary Medicine with 154 articles respectively.The research hotspots mainly include pneumonia,pulmonary infections,clinical efficacy,bronchoalveolar lavage fluid,bronchiectasis,and mechanical ventilation.Conclusion Fibrobronchial BAL is widely used in the diagnosis and treatment of respiratory diseases in China.In the future,it is necessary to strengthen the cooperation among researchers,deepen the theoretical basis,improve the clinical application system,and promote the further development of respiratory department in China.

Objective To explore the impact of artificial intelligence(AI)-assisted computing systems on the success rate of surgery,stereoscopic vision,and quality of life in patients with intermittent exotropia.Methods A total of 108 children(216 eyes)with intermittent exotropia who visited Ziyang Hospital of West China Hospital,Sichuan University from March 2021 to April 2023 were collected and randomly divided into Group A and Group B,with 54 children(108 eyes)in each group.Group A used the traditional calculation method,while Group B used the AI-assisted calculation method to determine the surgical volume of strabismus in children by the angles of strabismus at 6m and 33 cm.The success rate of surgery,postoperative angle of strabismus,stereoscopic visual function,horizontal and vertical perceptual eye positions,and quality of life[Child Intermittent Exotropia Questionnaire(CIXTQ)score]were compared between the two groups at 1 day,1 month,3 months and 6 months after surgery.Results At 1 day,1 month,3 months and 6 months after surgery,the success rate of surgery in Group B was significantly higher than that in Group A(all P＜0.05).Compared with the val-ues at 1 day after surgery,the angles of strabismus at 6 m and 33 cm in the two groups were significantly reduced at 1 month,3 months and 6 months after surgery(all P＜0.05);Group B had significantly lower angles of strabismus at 6 m and 33 cm than Group A at 1 day,1 month,3 months and 6 months after surgery(all P＜0.05).At 1 month,3 months and 6 months after surgery,the central and far stereoscopic visions of children in the two groups gradually increased,with the increase rate in Group B being significantly higher than that in Group A(Finter-group=76.59,P＜0.05);the proportion of children without stereoscopic vision in both groups gradually decreased,with a decrease rate in Group B being significantly higher than that in Group A(Finter-group=137.74,P＜0.05).At 1 month,3 months and 6 months after surgery,the horizon-tal and vertical perceptual eye positions in Group B showed a significant descend range compared with those in Group A(all P＜0.05).At 1 day,1 month,3 months and 6 months after surgery,the exotropia displacement of children in both groups gradually increased,and the displacement of children in Group B was significantly smaller than that in Group A at 1 month,3 months and 6 months postoperatively(all P＜0.05).At 1 month,3 months and 6 months after surgery,the CIX-TQ scores of children in the two groups increased gradually,with CIXTQ scores in Group B being significantly higher than those of Group A(all P＜0.05).Conclusion AI-assisted computing systems can effectively improve the success rate of intermittent exotropia surgery,enhance the establishment of stereoscopic visual function,improve visual quality,and sig-nificantly improve the quality of life of children.

AIM:To evaluate the effectiveness of heparin-binding protein(HBP)in combination with organ function indicators for early diagnosis and prognosis prediction in patients with severe trauma complicated with sepsis.METHODS:A retrospective analysis was conducted on 184 patients with multiple injuries who were admitted to the Emergency Medicine Department of the Second Affiliated Hospital of Zhejiang University Medical College between January 2019 and September 2020 and underwent HBP testing.Patients were classified according to the SEPSIS 3.0 diagnostic cri-teria into a sepsis group(n=89)and a non-sepsis group(n=95).Clinical outcomes were tracked,dividing patients into a deceased group(n=43)and a survival group(n=141).HBP levels were continuously measured,and the peak values of the two groups were compared to assess the efficacy of diagnosing sepsis.Further analysis on the correlation of HBP peak value median with clinical prognosis was conducted.The effectiveness of HBP alone and in combination with total biliru-bin(TBil)and white blood cell(WBC)count in prognosis assessment was evaluated.RESULTS:(1)No significant dif-ference was found in the peak level of HBP between the sepsis group(n=89)and the non-sepsis group(n=95)(71.7±68.6 vs 52.5±56.1,P=0.051).(2)Among the 184 patients,the peak level of HBP was positively correlated with WBC count(r=0.244,P<0.01)and TBil levels(r=0.241,P<0.01).(3)The area under curve(AUC)for independent diag-nosis of sepsis using TBil levels,WBC count,and PCT levels were 0.618,0.631,and 0.718,respectively,and the com-bined AUC was 0.684,with a diagnostic sensitivity of 60.7%and specificity of 71.6%(P<0.05).(4)Prognostic analy-sis of mortality showed that patients in the high HBP level group had a significantly higher mortality rate than those in the low-level group(30.4%vs 16.3%,P<0.05).The WBC count was also significantly higher in the deceased group than in the survival group(17.5±6.9 vs 12.8±4.7,P<0.01),especially in those with sepsis(P<0.01).The AUCs for predict-ing sepsis mortality prognosis using HBP peak level,TBil levels,WBC count,SOFA score,and APACHE-II score were 0.618,0.603,0.719,0.823,and 0.811,respectively.The combined AUC of HBP with TBil and WBC for assessing sepsis prognosis was 0.750,with a sensitivity of 74.4%and specificity of 74.5%,showing statistically significant differ-ences(P<0.05).(5)The combined assessment of these three indicators showed no statistically significant difference from artificial scoring systems in predicting sepsis prognosis(P>0.05).CONCLUSION:The combination of HBP,TBil,and WBC is highly effective in predicting the risk of sepsis in patients with multiple injuries and has significant clinical value in predicting the mortality risk of trauma patients with sepsis.

Objective Visual electrophysiology and optical correlation tomography angiography(OCTA)were used to investigate differences in preoperative retinal function in patients with type 2 diabetic cataract(DC)without obvious retinopathy(NDR)and to determine the clinical application of whole-process blood glucose man-agement(WBGM)for improving postoperative vision in DC patients.Methods This study investigated the preop-erative and postoperative visual electrophysiology(N75,P100,photopic FERG,and scotopic FERG),peripapil-lary retinal nerve fiber layer(pRNFL)and peripapillary capillary vessel density(ppVD)data,as well as trends in these data changes during blood glucose management intervention.Results As the course of T2DM progressed,FBG and HbA1c increased,the N75 and P100 lategraduancy periods of patients gradually lengthened,and the photopic FERG,scotopic FERG,pRNFL,and ppVD values decreased at each postoperative time point.At the same time,the best corrected visual acuity(BCVA)of patients after surgery gradually decreased(P<0.05).Compared with that at 1 week after surgery,the BCVA of Group A(without whole-process blood glucose manage-ment)gradually increased at 1 month and 3 months after surgery but showed a downward trend at 1 year after sur-gery.The BCVA of Group B(with whole-process blood glucose management)gradually stabilized at 1 month after surgery,and at all time points after surgery,the BCVA of Group B was better than that of Group A.The results showed that N75 and P100 in Group A were greater than those in Group B,while the photopic and scotopic FERG,pRNFL,and ppVD(%)in Group A were lower than those in Group B.In addition,N75 and P100 in Group A showed a gradual prolongation trend at various time points after surgery,while photopic FERG,scotopic FERG,pRNFL,and ppVD(%)showed a gradually decreasing trend.However,the above data in Group B gradu-ally stabilized at 3 months after DC surgery,approaching the preoperative level of the group(P<0.05).In the state of whole blood glucose management,although N75 and P100 both reached their longest durations at 1 week af-ter surgery,N75,P100,photopic FERG,scotopic FERG,and pRNFL showed a gradually decreasing trend at 1 month and 3 months after surgery,while ppVD(%)gradually increased(P<0.05).At various time points from 3 months to 1 year after surgery,the overall trend of the above indicators remained stable and close to the preoperative values(P>0.05).Conclusion According to the results of the quantitative analysis of visual electrophysiology and OCTA,in DC patients without obvious retinopathy and with the worsening of diabetes,retinal function decreased,but whole-process blood glucose management effectively restored retinal function and improved visual quality after surgery.

Objective:To establish a method via transfection of RNA to rescue coxsackievirus B3 B3 (CVB3).Methods:The efficiency of CVB3 genomic RNA extraction from three nucleic acid extraction reagents, Qiagen 57704, Qiagen 52904, and Trizol, and the transfection efficiency of viral RNA with two transfection reagents (Lipofectamine 2000 and Lipofectamine 3000) were compared. The efficiency of CVB3 rescue in Vero cells and HEK293T cells to determine the optimal conditions for rescuing CVB3.Results:The number of phagolysosomes for virus rescue by Qiagen 57704, Qiagen 52904, and Tizol kit extracted RNA was 13.33±1.53, 150±15.00, and 1.67±0.58, respectively, and there was a statistically significant difference in the efficiency of the three method of extracting CVB3 RNA to rescue the viral RNA ( F=268.920, P<0.001); The number of phage spots formed by Lipofectamine3000 and Lipofectamine2000 transfected RNA was 74.50±3.00 and 22.00±5.00, respectively, and the difference was statistically significant ( P<0.01); Qiagen 52904 reagent extracted CVB3 nucleic acid more efficiently than Qiagen 57704 and Trizol reagents; the transfection efficiency of transfection reagent Lipofectamine 3000 was 3 times more than than that of Lipofectamine 2000, and the efficiency of virus rescue of CVB3 in HEK293T cell culture was higher than that of HeLa and Vero cells, and the copy numbers of the three kinds of viruses rescuing the virus were 6.09×10 7±8.00×10 5, 5.18×10 3±6.17×10 2 and 0, the difference was statistically significant ( F=17 383.644, P<0.001), and it was also found that the efficiency of virus rescue could be improved by multiple elution when extracting RNA. Conclusions:In this study, we successfully established the method of transfecting RNA to rescue CVB3, which can effectively improve the efficiency of virus rescue by choosing Qiagen 52904 nucleic acid extraction kit, increasing the number of elution, using Lipofectamine 3000 transfection reagent, and transfection of HEK293T cells.