ObjectiveTo validate the efficacy of Yunpi Huatan Tongqiao prescription （YHTP） in down-regulating glycolysis to modulate microglia phenotype and improve inflammation and cognitive memory deficits in obstructive sleep apnea （OSA） mice. MethodForty-eight male Balb/C mice were randomly divided into a normal group， a model group， a montelukast sodium group （30 mg·kg^-1）， and low， medium， and high dose groups of YHTP （8.28， 16.56， and 33.12 g·kg^-1）， with 8 mice in each group. All groups， except the normal group， received intraperitoneal injections of lipopolysaccharide （LPS） and underwent chronic intermittent hypoxia （CIH） modeling for 4 weeks. Subsequently， the mice were treated with medications for 4 weeks and then sampled. Animal behavioral tests assessed memory impairment due to hypoxia. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to measure mRNA expression levels of M1-associated inflammatory factors interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and markers such as T lymphocyte activation antigen （CD86） and inducible nitric oxide synthase （iNOS）， as well as M2-associated inflammatory factors interleukin-10 （IL-10）， transforming growth factor-β （TGF-β）， and the marker mannose receptor （CD206） in hippocampal tissue. Western blot was employed to detect differences in the expression of M1 and M2 microglia phenotypic markers （CD86， CD206） and glycolysis-related proteins glucose transporter type 1 （GLUT1）， hexokinase 2 （HK2）， phosphofructokinase （PFKM）， pyruvate kinase 2 （PKM2）， and monocarboxylic acid transporter 1 （MCT1）. ResultBehavioral tests showed that compared to the results in the normal group， the Y-maze autonomous alternation rate was significantly reduced in the model group （P<0.01）. The latency time for the target hole in the Barnes' maze during the training period （days 2， 3， 4） and testing period （days 5， 12） was significantly increased （P<0.05， P<0.01）. M1 glial cell markers CD86 and iNOS， as well as inflammatory factors IL-1β and TNF-α mRNA， were significantly elevated （P<0.01）. In contrast， the mRNA expression of M2 glial cell markers IL-10， CD206， and TGF-β was significantly reduced （P<0.01）. The protein expression of glycolytic proteins HK2， PFKM， PKM2， MCT1， and the M1 marker CD86 was significantly increased （P<0.05， P<0.01）， while M2 marker CD206 protein expression was significantly decreased （P<0.01）. Compared to the results in the model group， the Y-maze autonomous alternation rate was significantly increased in the medium and high dose groups of YHTP （P<0.05， P<0.01）. The latency time for the target hole during the training （day 4） and testing periods （days 5， 12） was significantly reduced （P<0.01）. Real-time PCR results indicated that mRNA expression levels of M1-related pro-inflammatory factors in the hippocampal tissue were significantly reduced in the low， medium， and high dose groups of YHTP （P<0.01）， while M2-related inflammatory factors' mRNA expression was significantly increased （P<0.01）. Western blot results showed that in the medium and high dose groups of YHTP， the expression of the M1 marker CD86 in the hippocampus was reduced， whereas the expression of the M2 marker CD206 was significantly increased （P<0.01）， with a significant decrease in the expression of glycolysis-related proteins （P<0.01）. ConclusionYHTP can improve inflammation and cognitive impairment induced by hypoxia in OSA model mice. This is achieved by downregulating glycolysis in brain microglia， inhibiting M1 activation， reducing pro-inflammatory factor release， and promoting M2 activation， thereby exerting a therapeutic effect on inflammation and cognitive impairment caused by OSA.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective To construct and apply a multidrug-resistant bacteria information management system,and evaluate its effectiveness and accuracy in the management of patients with multidrug-resistant bacteria infection.Methods A system construction team was established to develop a multidrug-resistant bacteria information management system,which includes 4 modules:early warning,execution,monitoring,and statistical decision-making.Patients with positive detection of multidrug-resistant bacteria admitted to a tertiary A obstetrics and gynecology hospital in Zhejiang Province from January to December 2022,members of the system construction team,and clinical medical staff were selected as the research subjects.The execution efficiency of multidrug-resistant bacteria management,as well as the system's accuracy,usability,and satisfaction were compared before(January to June 2022)and after(July to December 2022)the application of the system.Results After the implementation of the system,the immediate feedback rate of multidrug-resistant bacteria early warning information increased from 62.87％to 89.78％;the rate of issuing isolation medical orders rose from 61.07％to 93.33％;the accuracy of the implementation of isolation measures for patients increased from 66.67％to 98.01％;all differences are statistically significant(P＜0.001).The accuracy rate of the system in making decisions is 88.44％;the usability score given by medical staff for the system is 44.04 points,and the satisfaction score of the system construction team members and medical staff to the system is 121.25 points,both at a high level.Conclusion The multidrug-resistant information management system is equipped with features such as immediate alerts,multi-channel notifications,infection control department supervision,and auxiliary decision-making,which can provide medical staff with accurate decision reports.Preliminary application results show that the system has a high level of accuracy and good usability.

Clinical cases treated by magnetic compression anastomosis (MCA) for different causes and types of intestinal stenosis/ atresia to successfully achieve intestinal recanalization were reviewed, so as to explore the clinical application of MCA. From May 2019 to August 2022, 4 patients underwent colorectal MCA for intestinal recanalization in the First Affiliated Hospital of Xi'an Jiaotong University and Northwest Women and Children's Hospital. All operations went well, and the intestinal anastomosis was recanalized. The magnetic ring was discharged in 7-15 days, and the postoperative colonoscopy or radiography showed that the anastomosis was intact. MCA can be used to treat different types of colorectal stenosis and atresia due to different reasons, and can also be used to assist intestinal anastomosis in colorectal surgery.

Objective    To evaluate the clinical value of in vitro fenestration and branch stent repair in the treatment of thoracoabdominal aortic aneurysm in visceral artery area assisted by 3D printing. Methods    The clinical data of 7 patients with thoracoabdominal aortic aneurysm involving visceral artery at the Department of Vascular Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University from March 2016 to May 2019 were analyzed retrospectively. There were 5 males and 2 females with an average age of 70.2±3.9 years. Among them 4 patients had near-renal abdominal aortic aneurysm, 3 had thoracic aortic aneurysm, 4 had asymptomatic aneurysm, 2 had acute symptomatic aneurysm and 1 had threatened rupture of aneurysm. According to the preoperative CT measurement and 3D printing model, fenestration technique was used with Cook Zenith thoracic aortic stents, and branch stents were sewed on the main stents in vitro, and then the stents were modified by beam diameter technique for intracavitary treatment. Results    All the 7 patients completed the operation successfully, and a total of 18 branch arteries were reconstructed. The success rate of surgical instrument release was 100.0%. The average operation time was 267.0±38.5 min, the average intraoperative blood loss was 361.0±87.4 mL and the average hospital stay was 16.0±4.2 d. Immediate intraoperative angiography showed that the aneurysms were isolated, and the visceral arteries were unobstructed. Till May 2019, there was no death, stent displacement, stent occlusion, ruptured aneurysm or loss of visceral artery branches. Conclusion    3D printing technology can completely copy the shape of human artery, intuitively present the anatomical structure and position of each branch of the artery, so that the fenestration technique is more accurate and the treatment scheme is more optimized.

The count and recognition of white blood cells in blood smear images play an important role in the diagnosis of blood diseases including leukemia. Traditional manual test results are easily disturbed by many factors. It is necessary to develop an automatic leukocyte analysis system to provide doctors with auxiliary diagnosis, and blood leukocyte segmentation is the basis of automatic analysis. In this paper, we improved the U-Net model and proposed a segmentation algorithm of leukocyte image based on dual path and atrous spatial pyramid pooling. Firstly, the dual path network was introduced into the feature encoder to extract multi-scale leukocyte features, and the atrous spatial pyramid pooling was used to enhance the feature extraction ability of the network. Then the feature decoder composed of convolution and deconvolution was used to restore the segmented target to the original image size to realize the pixel level segmentation of blood leukocytes. Finally, qualitative and quantitative experiments were carried out on three leukocyte data sets to verify the effectiveness of the algorithm. The results showed that compared with other representative algorithms, the proposed blood leukocyte segmentation algorithm had better segmentation results, and the mIoU value could reach more than 0.97. It is hoped that the method could be conducive to the automatic auxiliary diagnosis of blood diseases in the future.
Algorithms ; Leukocytes

OBJECTIVES: Nasopharyngeal carcinoma (NPC) is a highly invasive epithelial malignant tumor with unique geographical and ethnic distribution characteristics. NPC is mostly found in south China and Southeast Asia, and its treatment mainly depends on radiotherapy and chemotherapy. However, NPC is usually found in the late stage, and local recurrence and distant metastasis are common, leading to poor prognosis. The receptor tyrosine kinase AXL is up-regulated in various tumors and it is involved in tumor proliferation, migration, invasion, and other processes, which are associated with poor prognosis of tumors. This study aims to detect the expression of AXL in NPC cell lines and tissues, and to investigate its biological function of AXL and the underlying molecular mechanisms in regulation of NPC. METHODS: The expression levels of AXL in normal nasopharyngeal epithelial tissues and NPC tissues were analyzed by GSE68799, GSE12452, and GSE53819 data sets based on Gene Expression Omnibus (GEO) database. The Cancer Genome Atlas (TCGA) database was used to analyze the relationship between AXL and prognosis of head and neck squamous cell carcinoma (HNSC). The indicators of prognosis included overall survival (OS), disease-free interval (DFI), disease-specific survival (DSS), and progression-free interval (PFI). Western blotting assay was used to detect the AXL protein expression levels in normal nasopharyngeal epithelial cell line and NPC cell lines. Immunohistochemical method was used to detect AXL expression levels in normal nasopharyngeal epithelial tissues and NPC tissues. Cell lines with stable AXL knockdown were established by infecting 5-8F and Fadu cells with lentivirus interference vector, and cell lines with stable AXL overexpression were established by infecting C666-1 and HK-1 cells with lentivirus expression vector. Real-time PCR and Western blotting were used to detect the efficiency of knockdown and overexpression in stable cell lines. The effects of AXL knockdown or overexpression on proliferation, migration, and invasion of NPC cells were detected by CCK-8, plate colony formation, and Transwell assays, and the effect of AXL knockdown on tumor growth in nude mice was detected by subcutaneous tumor formation assay. The sequence of AXL upstream 2.0 kb promoter region was obtained by UCSC online database. The PROMO online database was used to predict AXL transcription factors with 0% fault tolerance, and the JASPAR online database was used to predict the binding sites of ETS1 to AXL. Real-time PCR and Western blotting were used to detect the effect of ETS1 on AXL protein and mRNA expression. The AXL upstream 2.0 kb promoter region was divided into 8 fragments, each of which was 250 bp in length. Primers were designed for 8 fragments. The binding of ETS1 to AXL promoter region was detected by chromatin immuno-precipitation (ChIP) assay to determine the direct regulatory relationship between ETS1 and AXL. Rescue assay was used to determine whether ETS1 affected the proliferation, migration, and invasion of NPC cells through AXL. RESULTS: Bioinformatics analysis showed that AXL was highly expressed in NPC tissues (P<0.05), and AXL expression was positively correlated with OS, DFI, DSS, and PFI in HNSC patients. Western blotting and immunohistochemical results showed that AXL was highly expressed in NPC cell lines and tissues compared with the normal nasopharyngeal epithelial cell line and tissues. Real-time PCR and Western blotting results showed that knockdown and overexpression efficiency in the stable cell lines met the requirements of subsequent experiments. The results of CCK-8, plate colony formation, Transwell assays and subcutaneous tumor formation in nude mice showed that down-regulation of AXL significantly inhibited the proliferation, migration, invasion of NPC cells and tumor growth (all P<0.05), and the up-regulation of AXL significantly promoted the proliferation, migration, and invasion of NPC cells (all P<0.05).As predicted by PROMO and JASPAR online databases, ETS1 was a transcription factor of AXL and had multiple binding sites in the AXL promoter region. Real-time PCR and Western blotting results showed that knockdown or overexpression of ETS1 down-regulated or up-regulated AXL protein and mRNA expression levels. ChIP assay result showed that ETS1 bound to AXL promoter region and directly regulate AXL expression. Rescue assay showed that AXL rescued the effects of ETS1 on proliferation, migration and invasion of NPC cells (P<0.05). CONCLUSIONS AXL is highly expressed in NPC cell lines and tissues, which can promote the malignant progression of NPC, and its expression is regulated by transcription factor ETS1.
Animals ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Gene Expression Regulation, Neoplastic ; Mice ; Mice, Nude ; Nasopharyngeal Carcinoma/genetics* ; Nasopharyngeal Neoplasms/metabolism* ; RNA, Messenger/genetics* ; Sincalide/metabolism* ; Transcription Factors/genetics*

Objective:To investigate the clinical characteristics of influenza A and influenza B pneumonia and the risk factors of severe influenza pneumonia in children.Methods:The epidemiology, clinical characteristics, laboratory tests and pathogens of co-infection in children with pneumonia caused by influenza A virus and influenza B virus, and the risk factors of severe influenza pneumonia were retrospectively analyzed.Results:(1) The cases of influenza A infection accounted for 65.1% and those with influenza B infection accounted for 32.9% among the 711 children with influenza pneumonia.The dominant strain was Influenza B Victoria virus in spring and summer, influenza A(H 3N 2) virus in autumn, and influenza A(H1N1) virus in winter.The dominant strain was influenza A virus at the age of < 1 year and ~3 years, influenza A virus and influenza B virus at the age of ~6 years, and influenza B virus at the age of ≥6 years.(2) The gastrointestinal symptoms were more common in children with influenza B pneumonia compared with those with influenza A pneumonia(53.4% vs 44.7%, χ2=4.728, P=0.030), but crackles and wheezing were more common in children with influenza A pneumonia compared with those with influenza B pneumonia(80.1% vs 70.5%, 36.9% vs 25.6%, χ2=8.945, 8.093, all P<0.05). (3) The percentage of decreased lymphocyte count in children with influenza B pneumonia was higher than those with influenza A pneumonia(5.6% vs 1.9%, χ2=6.633, P=0.010). (4) Mixed Mycoplasma Pneumoniae was more common in children with influenza B pneumonia compared with those with influenza A pneumonia(23.9% vs 10.8%, χ2=20.789, P<0.001), and mixed virus and bacteria were more common in children with influenza A pneumonia compared with those with influenza B pneumonia(15.8% vs 8.1%, 50.1% vs 41.9%, χ2=7.934, 4.221, all P<0.05). (5) Multivariate logistic regression analysis showed that age <2 years( OR=1.886, 95% CI 1.149~3.096, P=0.012), increased LDH( OR=1.736, 95% CI 1.080~2.790, P=0.023), the percentage of lymphocyte decreased( OR=2.762, 95% CI 1.669~4.571, P<0.001) and the percentage of CD3 + decreased ( OR=6.019, 95% CI 3.993~9.331, P<0.001)were risk factors for severe influenza pneumonia. Conclusion:Among hospitalized children with influenza pneumonia, there were some differences in the age of infection, clinical characteristics, laboratory tests and pathogens of co-infection between the cases caused by influenza B and influenza A, and clinicians should remain vigilant for the occurrence of severe influenza pneumonia.

Objective:To retrospectively analyze the efficacy and safety of dupilumab in the treatment of bullous pemphigoid (BP) .Methods:Clinical data were collected from BP patients who received injections of dupilumab at an initial dose of 600 mg followed by an every-2-week regimen at a dose of 300 mg (the frequency of injections could be increased if necessary) in Department of Dermatology, Peking University First Hospital from October 2020 to October 2021, and their clinical manifestations and changes in laboratory indices were analyzed.Results:A total of 21 BP patients treated with dupilumab were included in this study. Nineteen (90.5%) patients achieved complete or marked disease control after 2-week treatment with dupilumab; 12 patients were followed up for 16 weeks, and all maintained complete disease control at 16 weeks. All patients had a bullous pemphigoid disease area index (BPDAI) score of 122.5 ± 51.1 points at baseline, which decreased to 30.6 ± 27.4 points after 2-week treatment with dupilumab ( t = 8.53, P < 0.001) , and continued to decrease to 12.7 ± 9.1 points after 4-week treatment ( t = 9.73, P < 0.001) . Pruritus was markedly relieved in all the 21 patients within 4-week treatment with dupilumab. Among 10 patients with elevated eosinophil counts at baseline, the eosinophil counts markedly decreased in 9 after treatment. The serum IgE level was elevated in 7 patients at baseline, which markedly decreased in 6 after treatment. Viral conjunctivitis occurred in 1 (4.8%) patient, and no adverse reactions were observed in other patients. Conclusion:Dupilumab is effective in the control of BP and relief of pruritus, with a favorable safety profile.

Objective:To investigate the detection, epidemiological and clinical characteristics of human rhinovirus(HRV) in hospitalized children with respiratory tract infections.Methods:The study population comprised of 10 514 children with respiratory tract infections admitted to Department of Respiration, the Children′s Hospital of Soochow University, between January 2013 and December 2019.The nasopharyngeal aspirates and medical history were obtained by qualified medical personnel.Reverse transcription-polymerase chain reaction method was used to test HRV.Results:The total positive rate of human rhinovirus was 14.2%(1 493/10 514), and there was no significant difference between male and female( χ2=2.006, P=0.157). The positive rates from 2013 to 2019 were 9.7%, 14.6%, 19.1%, 18.6%, 18.1%, 11.0%, 11.4% respectively, and there were significant differences among these groups( χ2=116.580, P<0.001). HRV distributed throughout the year with a peak in summer and autumn(June to November), followed by spring, and the lowest in winter.The detection rates of HRV infection rates were 14.2%, 15.5%, 13.5% and 9.8% in the age group of 28 d~6 months, ~2 years, ~7 years and>7 years respectively, and there were significant differences among these age groups( χ2=16.124, P<0.001). The detection rate of HRV in children under 2 years was higher( χ2=7.711, P=0.005). The clinical characteristics of HRV infection were fever, cough, wheezing and even dyspnea.Bronchopneumonia had the highest percentage(68.9%), followed by bronchitis(13.2%). Compared with non-coinfection group, patients with coinfection with other viruses were more prone to wheezing and pulmonary rales( χ2=9.483, 10.821, P=0.024, 0.013), and coinfection with mycoplasma was more likely to cause fever and lobar pneumonia( χ2=51.585、96.060, P all<0.001); 57.8% presented leukocytosis, while 15.6% showed a higher CRP(>15 mg/ml). The increase of CRP and leukocytosis were more obvious in children under 2 years of age( χ2=26.097, 55.973, P all<0.001). Conclusion:HRV was a major viral pathogen of RTIs in recent 7 years, distributing throughout the year with a peak in summer and autumn, mainly involving children under 2 years of age.The clinical features were diverse, and the clinical symptoms were severe in childhood coinfections with other pathogens.