Objective:To explore the application value of high-resolution temporal bone CT and DW-MRI fusion technology in achieving precise diagnosis and anatomical localization of middle ear cholesteatoma during endoscopic surgery. Methods:Eighteen patients initially diagnosed with middle ear cholesteatoma in the Department of Otolaryngology Head and Neck Surgery, Shaanxi Provincial People's Hospital, from January to June 2024 were enrolled.Preoperative high-resolution temporal bone CT and DW-MRI were performed, and rtStation software was used for image fusion to construct CT-MRI fused images. The involvement of cholesteatoma in six anatomical subregions of the temporal bone was evaluated. Using surgical pathology as the gold standard, and combining surgical videos and anatomical records, the sensitivity, specificity, and accuracy of pure CT, pure DW-MRI, and CT-MRI fused images in evaluating middle ear cholesteatoma lesions were compared. Results:A total of 18 patients were included, and 17 cases were pathologically confirmed as middle ear cholesteatoma postoperatively. The sensitivity of the preoperative of preoperative CT was 100%, but the specificity was only 44.44%, with an overall accuracy of 72.22%; the sensitivity and specificity of DW-MRI evaluation were 81.46% and 85.19%, the accuracy was 83.33%, respectively. In contrast, the sensitivity and specificity of CT-MRI fusion image to the spatial localization of cholesteatoma were higher than that of DW-MRI alone（92.59% vs 81.46%; 98.15% vs 85.19%）, and the diagnostic accuracy was also significantly improved（95.37% vs 83.33%）. The Kappa values for the agreement between HRCT, DW-MRI, and CT-MRI segmentation localization and pathological results were 0.444, 0.667, and 0.907 respectively. The chi-square paired t-test confirmed statistically significant diagnostic differences between groups（P<0.001）. Results demonstrated that CT-MRI significantly outperformed HRCT and DW-MRI in diagnostic efficacy for segmental localization of primary posterior congenital middle ear cholesteatoma. Conclusion:High-resolution temporal bone CT combined with DW-MRI fusion technology demonstrates higher sensitivity, specificity, and accuracy in the diagnosis and spatial localization of middle ear cholesteatoma than single imaging modalities. It can provide more precise evaluation of lesion scope for endoscopic surgery, showing important clinical application value.
Humans ; Cholesteatoma, Middle Ear/diagnostic imaging* ; Tomography, X-Ray Computed ; Temporal Bone/diagnostic imaging* ; Diffusion Magnetic Resonance Imaging ; Female ; Male ; Adult ; Sensitivity and Specificity ; Middle Aged ; Endoscopy

Objective To prepare humanized monoclonal antibodies(Mabs)targeting Acinetobacter baumannii(Ab)based on single memory B cell sequencing technology,construct the immune repertoire of the core protein of Ab,hemolysin-coregulated protein(Hcp),and express its Mabs with binding activity.Methods E.coli BL21 harboring the recombinant plasmid pGEX-6p-1-Hcp was constructed.Hcp protein was obtained using protein expression and affinity chromatography.Female SPF BALB/c mice(6～8 weeks old,weighing 18～20 g)were immunized intramuscularly with antigen Hcp to generate specific memory B cells.Single antigen-specific memory B cells were sorted using flow cytometry.The immune repertoire of Hcp was constructed using single-cell sequencing technology,and bioinformatics analysis was performed on the sequencing results.Mabs were obtained using antibody humanization techniques.The in vitro binding activity of the antibodies was detected by ELISA.Results The target protein Hcp with a purity>95%was obtained after expression and purification.The immune repertoire of Hcp was successfully constructed,and the results of BCR clonotype identification and analysis,CDR3 region characteristic analysis,and V-J gene pairing characteristic analysis were achieved.Antibody humanization got 7 Mabs,that is,IgG1-1,IgG1-2,IgG2-1,IgG2-2,IgG3-1,IgG4-1 and IgG4-2.ELISA results showed IgG1-1,IgG3-1,IgG4-1,and IgG4-2 had an antibody binding titer of 1∶1 280,IgG2-2 of 1∶10 240,IgG2-1 of 1∶5 120,and IgG1-2 of 1∶160.Conclusion Single-cell sequencing technology enables rapid,accurate,and efficient construction of an Hcp protein immune repertoire containing extensive antibody information.Utilizing this immune repertoire allows for the expression of Mabs with binding activity.

Objective:To compare the effects of different test meals on postprandial triglycerides and to optimize the standard meal composition and the blood sampling protocol for the oral fat tolerance test.Methods:This study is a prospective, open-label, randomized, cross-over trial. In March 2023, 36 volunteers were recruited in Hebei General Hospital. They underwent a health examination and oral glucose tolerance test. Twenty-six healthy volunteers(11 males and 15 females) were included in this study, with an average age of(39.08±4.56) years. Each volunteer received 75 g protein meal, 75 g fat meal, 700 kcal fixed-calorie high-fat mixed meal, and a high-fat mixed meal with energy adjusted based on 10 kcal/kg body weight. A one-week washout period of regular diet was applied before each trial. Blood was collected at fasting status and 1, 2, 3, 4, 5, and 6 hours after a meal to detect serum triglycerides, total cholesterol, low density lipoprotein-cholesterol(LDL-C), high density lipoprotein-cholesterol(HDL-C), glucose, and insulin. The variations of postprandial metabolic indicators over time following the consumption of different test meals were analyzed. The disparities in postprandial metabolic responses between the two types of mixed meals were compared.Results:The protein meal, fat meal, fixed-calorie high-fat mixed meal, and adjusted-calorie high-fat mixed meal resulted in postprandial triglyceride increases of 22.45%, 115.40%, 77.14%, and 63.63%, and insulin increase of 560.43%, 85.69%, 554.18%, and 598.97%, respectively, and with reductions in total cholesterol, LDL-C, and HDL-C ranging from 5.64%-21.81%, respectively. The blood glucose changed slightly. Changes in metabolic indicators mainly occured within 4 hours. The comparison of the characteristics of postprandial triglycerides between the two high-fat mixed meals showed no statistically significant differences( P>0.05). Conclusion:A standardize protocol with a 700 kcal fixed-calorie high-fat mixed meal as test meal, and blood lipid levels measured at fasting and at 1, 2, 3, and 4 hours after consumption, can serve as an optimized approach for oral fat tolerance test.

BACKGROUND:As a dominant breed pig in southwest China,the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines,but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE:To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS:Twenty adult female Yunnan small-ear pigs were equally randomized into two groups.In the autologous Achilles tendon group,the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft,while in the sham-operated group,a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament.General conditions of each pig were observed and recorded before and 12 months after surgery.Ligaments and grafts were taken for gross observation and MAS scoring.Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments.The staining and arrangement of type I and type Ⅲ collagen were evaluated by immunohistochemistry.Transmission electron microscopy was used to observe the type,size,diameter,ratio,and distribution of collagen fibers in ligaments. RESULTS AND CONCLUSION:All animals had normal diet and activity,good wound healing,no obvious inflammatory reaction,no local purulent infection,and no significant changes in mental and urinary conditions compared with those before surgery.The reconstructed cruciate ligament of the knee was intact,with no stiffness and normal range of motion.Both the anterior drawer and Lachman tests were negative.Gross observation of the graft:12 months after surgery,the grafts was in good position,with good integrity,obvious tension,ligament color close to the original anterior cruciate ligament,and complete surface synovial coverage.Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS Ⅱ type.In the sham-operated group,the intraarticular ligament was defined as MAS I type.Hematoxylin-eosin staining indicated that,12 months after surgery,collagen fibers in the autologous Achilles tendon group began to appear bundled,isotropic,and uniformly arranged,with more obvious isotropic corrugations,and the nuclei were mainly linear or spindle-shaped,which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group.Immunohistochemistry results indicated that,12 months after surgery,there was a higher expression of type I collagen and significantly less expression of type Ⅲ collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group.The degree of type I and type Ⅲ staining was similar in the two groups.Under the transmission electron microscope,the diameter,arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery,indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery.Through a comprehensive evaluation of animal general conditions,ligament general view,MAS score,hematoxylin-eosin staining,immunohistochemistry,and transmission electron microscopy observation,we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs,with good morphological,histological and ultrastructural results.

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori(Hp)infection in mice.Methods L.paracasei TK1501 was cultured for 32 h at 37℃in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×107 CFU/mL.The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption,cation exchange chromatography and HPLC,and its stability and antibacterial activity were assessed.The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection,which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL.Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments,and gastric tissues of the mice were collected for HE staining.Results L.paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid,base,and organic solvents.In the in vitro experiment,the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus,Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract.In the mouse models,treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice.Conclusion L.paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.

Purpose To explore the predictive value of nomogram model for invasive breast cancer with axillary lymph node metastasis.Materials and Methods Retrospective analysis was made on 122 patients suspected to be breast cancer in the General Hospital of Ningxia Medical University from September 2020 to March 2022.According to whether there was axillary lymph node metastasis,all subjects were divided into 57 patients in the metastasis group and 65 patients in the non-metastasis group.All lesions were pathologically confirmed by surgery.The two groups received synthesis of magnetic resonance imaging(syMRI),dynamic contrast enhancement magnetic resonance imaging(DCE-MRI)and diffusion weighted imaging(DWI)scans.The syMRI parameters[including T1,T2,proton density(PD)],DCE-MRI time signal intensity curve,apparent diffusion coefficient(ADC)value of breast lesions were measured.Compared the difference of parameters between the two groups,and screened the independent risk factors of invasive breast cancer with axillary lymph node metastasis.Results Logistic regression results showed that Ki-67(OR=2.971,95%CI 1.306-6.762,P=0.009),lesion size(OR=1.652,95%CI 1.067-2.556,P=0.024),ADCratio(OR=1.685,95%CI 1.014-2.801,P=0.044),T2ratio(OR=3.015,95%CI 1.433-6.340,P=0.003),PDratio(OR=2.782,95%CI 1.471-5.262,P=0.002)were independent risk factors for invasive breast cancer with axillary lymph node metastasis.The comparison of the five models showed that the Logistic regression model had the best performance,with the area under curve of 0.729(95%CI 0.621-0.789),the accuracy,specificity and sensitivity were 70.65%,62.79%and 77.55%,respectively.The accuracy of the nomogram model was tested,and C-index=0.844,the accuracy of the nomogram model established was good,cut-off risk was 0.468,and the cut-off score was 143.50,which means that when the total score exceeds 143.50,the risk of axillary lymph node metastasis would be higher than 46.8%.Conclusion Nomogram model has a good predictive ability for invasive breast cancer patients with axillary lymph node metastasis.

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori(Hp)infection in mice.Methods L.paracasei TK1501 was cultured for 32 h at 37℃in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×107 CFU/mL.The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption,cation exchange chromatography and HPLC,and its stability and antibacterial activity were assessed.The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection,which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL.Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments,and gastric tissues of the mice were collected for HE staining.Results L.paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid,base,and organic solvents.In the in vitro experiment,the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus,Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract.In the mouse models,treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice.Conclusion L.paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.

This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7％NaCl,10％NaCl,3.5％ethanol,4.0％ethanol and pH5 stress(P＜0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P＜0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P＜0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.

Objective:To understand the monkeypox knowledge awareness, risk perception and vaccination intention in men who have sex with men (MSM) in five cities in northeast China.Methods:A cross-sectional study was conducted by using electronic questionnaire in MSM selected by convenience sampling in five cities in northeast China (Shenyang, Panjin, Changchun, Harbin and Jiamusi) from June 28 to July 8, 2023 by local centers for disease control and prevention and MSM communities. The sample size was estimated to be 220. Information about their demographics, monkeypox-related knowledge awareness, perceived concern about epidemic risk perception, and monkeypox vaccination intention were collected. Logistic regression model was used to analyze related factors for MSM's monkeypox vaccination intention.Results:In 355 MSM, 63.9% (227/355) had monkeypox vaccination intentions, and 55.5% (197/355) had high awareness of monkeypox related knowledge with a mean knowledge awareness score of 3.7±1.5. MSM with education level of high-school and above (a OR=1.93, 95% CI:1.01-3.69), higher knowledge awareness score (a OR=1.19, 95% CI:1.02-1.40) and higher risk perception of monkeypox infection (a OR=1.82, 95% CI:1.15-2.88), were more willing to receive monkeypox vaccination. The main reasons for willingness to receive monkeypox vaccine were preventing monkeypox (86.3%, 196/227) and worrying about appearance being affected (62.1%, 141/227). The main reasons for unwillingness for the vaccination included concerns about vaccine safety (53.1%, 68/128), clinical progression of AIDS being affected (46.1%, 59/128) and efficacy of antiretroviral therapy being affected (44.5%, 57/128). Conclusions:The levels of knowledge awareness and vaccine intentions still need to be improved among MSM in five cities of northeast China. It is necessary to improve the awareness of monkeypox and intention of monkeypox vaccination, promote protected sex behavior and self-assessment of infection risk, reduce vaccine hesitancy and increase monkeypox vaccination intention in MSM in 5 cities in northeast China.

Atherosclerosis is classified as"vessel bi"in traditional Chinese medicine,and cold evil is one of its important causes.Cold,having the properties of coagulation,stagnation,and contraction,tends to narrow the diameter of blood vessels and slow down blood flow,which can promote the pathophysiological process of atherosclerosis.At present,the etiological theory of vessel bi caused by cold evil has not been fully elaborated.Based on the explanations of"evening"in"pathogenic cold affecting in the evening",such as"ying","end of year","end of day",and"time of yang decline and yin exuberance",this paper proposes the viewpoint that the cold evil tends to affect ying-blood and cause vessel bi,and discusses its pathogenesis.From the perspective of the external environment,cold evil affecting the vessel is conditional,often occurring in winter and in the middle of the night.From the perspective of internal environment,the cold evil affects the vessel in a process which can be manifested in three stages:cold injury,cold stagnation,and cold congelation.The strength of yang qi is the factor that determine whether the cold evil can cause disease.Exploring the etiology and pathogenesis of atherosclerosis from the perspective of"pathogenic cold affecting in the evening"provides a new idea for further elucidating the pathogenesis of the disease and expanding its clinical treatment.