[摘 要] 目的：探讨环磷腺苷效应元件结合蛋白（CREB）对前列腺癌细胞恶性生物学行为的影响。方法：常规培养前列腺癌细胞PC3，用转染试剂将过表达对照质粒，CREB过表达质粒（CREB-oe)、敲减对照序列（si-NC）和si-CREB序列转染至PC3细胞，分为vector，CREB-oe、si-NC和si-CREB组。用划痕愈合实验、Transwell小室实验和流式细胞术分别检测各组细胞的迁移、侵袭能力和细胞周期情况。用CRISPR/cas9技术构建CREB敲除的PC3细胞，用PC3细胞移植瘤实验检测CREB敲除对移植瘤生长的影响。结果：在PC3细胞中成功地敲减或过表达了CREB（均P < 0.01），过表达或敲减CREB均可明显促进或抑制PC3细胞的迁移、侵袭能力（均P < 0.01），过表达CREB可促使PC3细胞进入S期，而敲减CREB表达则使PC3细胞阻滞于G1期（均P < 0.01）。成功地构建了CREB敲除PC3细胞，敲除CREB可明显抑制PC3细胞移植瘤的生长（P < 0.01）。结论：敲减或敲除CREB能够抑制PC3细胞的迁移和侵袭，并使其阻滞于G1期，进而抑制移植瘤的生长。

Objective: To explore the influencing factors for sarcopenia among elderly male patients with type 2 diabetes (T2DM), so as to provide the basis for the early prevention and treatment of sarcopenia. Methods: Male T2DM patients aged 60 and above admitted to Hangzhou First People's Hospital from January to December 2024 were selected as the study subjects. Demographic data, T2DM complications, and blood biochemical parameters were collected. Physical activity levels were assessed using the Chinese version of the International Physical Activity Questionnaire. The diagnosis of sarcopenia was made according to the diagnostic procedures and criteria established by the Asian Working Group for Sarcopenia in 2019. Factors affecting sarcopenia among elderly male patients with T2DM were analyzed using multivariable logistic regression model. Results: A total of 455 elderly male patients with T2DM were surveyed, with a mean age of (71.80±9.55) years. The predominant physical activity level was moderate with 226 cases accounting for 49.67%. The disease course of T2DM was mainly from 10-<20 years, with 229 cases accounting for 50.33%. There were 140 cases of T2DM complications, accounting for 30.77%. A total of 138 cases of sarcopenia were detected, with a prevalence of 30.33%. Multivariable logistic regression analysis showed that age (OR=1.077, 95%CI: 1.003~1.156), body mass index (<18.5kg/m2, OR=11.056, 95%CI: 3.343~36.547; 18.5~<25.0 kg/m2, OR=2.633, 95%CI: 1.420~4.881), physical activity level (low, OR=2.469, 95%CI: 1.421~4.292), chronic obstructive pulmonary disease (yes, OR=1.871, 95%CI: 1.091~3.206), T2DM complications (yes, OR=3.015, 95%CI: 1.516~6.001), glycated hemoglobin (≥7%, OR=2.822, 95%CI: 1.423~5.590) and albumin (OR=0.810, 95%CI: 0.662~0.991) were factors affecting sarcopenia among elderly male patients with T2DM (P<0.05). Conclusion Advanced age, body mass index <25.0 kg/m2, low physical activity level, chronic obstructive pulmonary disease, T2DM complications, high glycated hemoglobin and low albumin are associated with a higher risk of sarcopenia in elderly male patients with T2DM.

Objective To mine and analyze the routine blood test data of children with allergic rhinitis (AR), identify routine blood parameters related to childhood allergic rhinitis, establish an effective diagnostic model, and evaluate the performance of the model. Methods This study was a retrospective study of clinical cases. The experimental group comprised a total of 1110 children diagnosed with AR at the First Affiliated Hospital of Air Force Medical University during the period from December 12, 2020 to December 12, 2021, while the control group included 1109 children without a history of allergic rhinitis or other allergic diseases who underwent routine physical examinations during the same period. Information such as age, sex and routine blood test results was collected for all subjects. The levels of routine blood test indicators were compared between AR children and healthy children using comprehensive intelligent baseline analysis, with indicators of P≥0.05 excluded; variables were screened by Lasso regression. Binary Logistic regression was used to further evaluate the influence of multiple routine blood indexes on the results. Five kinds of machine model algorithms were used, namely extreme value gradient lift (XGBoost), logistic regression (LR), gradient lift decision tree (LGBMC), Random forest (RF) and adaptive lift algorithm (AdaBoost), to establish the diagnostic models. The receiver operating characteristic (ROC) curve was used to screen the optimal model. The best LightGBM algorithm was used to build an online patient risk assessment tool for clinical application. Results Statistically significant differences were observed between the AR group and the control group in the following routine blood test indicators: mean cellular hemoglobin concentration (MCHC), hemoglobin (HGB), absolute value of basophils (BASO), absolute value of eosinophils (EOS), large platelet ratio (P-LCR), mean platelet volume (MPV), platelet distribution width (PDW), platelet count (PLT), absolute values of leukocyte neutrophil (W-LCC), leukocyte monocyte (W-MCC), leukocyte lymphocyte (W-SCC), and age. Lasso regression identified these variables as important predictors, and binary Logistic regression further analyzed the significant influence of these variables on the results. The optimal machine learning algorithm LightGBM was used to establish a multi-index joint detection model. The model showed robust prediction performance in the training set, with AUC values of 0.8512 and 0.8103 in the internal validation set. Conclusion The identified routine blood parameters can be used as potential biomarkers for early diagnosis and risk assessment of AR, which can improve the accuracy and efficiency of diagnosis. The established model provides scientific basis for more accurate diagnostic tools and personalized prevention strategies. Future studies should prospectively validate these findings and explore their applicability in other related diseases.
Humans ; Male ; Female ; Rhinitis, Allergic/blood* ; Child ; Biomarkers/blood* ; Retrospective Studies ; Early Diagnosis ; Child, Preschool ; ROC Curve ; Logistic Models ; Hematologic Tests ; Algorithms ; Adolescent ; Machine Learning

Objective:To investigate the etiological distribution of severe community-acquired pneumonia (CAP) in children in Hebei Province.Methods:The nasopharyngeal swab samples and clinical data from 314 children with severe pneumonia were retrospectively analyzed between January 2014 and January 2016.Results:Among the 314 children, 298 (94.94%) showed positive result for pathogens in their nasopharyngeal swab samples, with 246 cases (78.34%) of multiple pathogens, predominantly mixed viruses and bacteria (206 cases, 65.61%). A total of 848 strains of pathogens were detected, including 483 strains (56.96%) of viruses, predominantly respiratory syncytial virus with 97 strains of subtype A and 86 strains of B. Bacteria and atypical pathogens ( Mycoplasma pneumoniae) had 365 strains (43.04%), mainly Streptococcus pneumoniae (220 strains) and Haemophilus influenzae (119 strains). The detection rate of pathogens was higher in children under one year of age ( χ2=21.389, P<0.001). There were no significant differences in different seasons, but the detection rates of respiratory syncytial virus A, respiratory syncytial virus B, and Streptococcus pneumoniae were higher in autumn, winter, and spring, respectively, with statistically significant differences ( χ2=22.205, P<0.001; χ2=37.874, P<0.001; χ2=11.380, P=0.009). Conclusions:Nasopharyngeal swab sample testing in children with severe pneumonia typically shows a coexistence of viral and bacterial pathogens, with detection rates varying among different age groups and seasons. Nasopharyngeal swab sample testing for pathogens provides valuable references for the identification of clinical pathogens.

ObjectiveTo explore the mechanism of Buzhong Yiqitang in ameliorating inflammatory injury in autoimmune thyroiditis (AIT) mice based on the Toll-like receptor 4（TLR4）/nuclear transcription factor-kappa B（NF-κB）/absent in melanoma 2（AIM2）inflammasome signaling pathway. MethodThe 120 genetically susceptible 8-week-old NOD.H-2^h4 mice were selected and randomly divided into control group， model group， low， medium and high dose groups of Buzhong Yiqitang （4.78， 9.56， 19.12 g·kg^-1）， and western medicine group （selenium yeast tablets， 3.033×10^-5 g·kg^-1）. The AIT model mice in each group drank ad libitum 0.05% sodium iodide aqueous solution for 8 weeks to establish the AIT model， and the control group drank ad libitum distilled water. Eight weeks later， the mice in each dosing group were divided into groups and gavage. The swelling of thyroid tissue was observed with the naked eye， and the weight of spleen was weighed. The content of serum inflammatory factors interleukin-1β （IL-1β）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α） was measured by enzyme-linked immunosorbent assay （ELISA）， and Real-time PCR was used to detect the expression of HMGB1， TLR4， AIM2， NF-κB p65，apoptosis-associated speck-like protein（ASC），cysteinyl aspartate-specific protease-1（Caspase-1）， IL-1β mRNA. Western blot was used to detect the expression of high motility group protein 1 （HMGB1）， TLR4， AIM2， NF-κB p65， phosphorylation（p）-NF-κB p65， ASC， Caspase-1， and IL-1β proteins in thyroid tissue， and immunofluorescence staining was used to observe the protein expression of HMGB1， AIM2， and NF-κB p65 in thyroid tissue of mice. ResultCompared with the control group， the thyroid tissue of mice in the model group was significantly swollen， the spleen quality was significantly increased， and the expression of HMGB1， TLR4， NF-κB p65， AIM2， ASC， Caspase-1， IL-1β in thyroid tissue was significantly increased （P<0.01）. Compared with the model group， the swelling of thyroid tissue in mice in each dose group of Buzhong Yiqitang was improved， the quality of spleen was significantly reduced， and the expression of HMGB1， TLR4， AIM2， NF-κB p65， p-NF-κB p65， ASC， Caspase-1， IL-1β in thyroid tissue was significantly reduced （P<0.05， P<0.01）. ConclusionBuzhong Yiqitang can effectively improve the inflammatory injury of AIT， and regulating the abnormal activation of the TLR4/NF-κB/AIM2 inflammasome signal pathway may be one of its intervention mechanisms.

Objectives To analyze the spatial and temporal aggregation of multidrug resistant pulmonary tuberculosis (MDR-TB) incidence in Nanning at the township / street scale from 2017 to 2021, to explore the spatial and temporal characteristics of the spread of MDR-TB in Nanning, and to provide a scientific reference basis for the health administrative departments to achieve the precise implementation of MDR-TB prevention and control. Methods Based on the data of MDR-TB cases in Nanning from 2017 to 2021, the spatial-temporal scanning analysis software SaTScan v9.7 was used to retrospectively detect and analyze the areas where MDR-TB cases gathered. Results Through simple spatial scanning analysis, it was found that there were three first-class aggregation areas (the aggregation center was Fujiayuan Street, Jiangnan District, 2017, Xinyang Street, Xixiangtang District, 2019, and Zhonghe Town, Yongning District, 2020), and one second-class aggregation area (the aggregation center was Jinchai Town, Mashan County, 2020). Simple time scanning showed that the clustering occurred from May 2019 to December 2020. Temporal and spatial aggregation analysis showed that Xinyang Street in Xixiangtang District was the center of the first-class aggregation area, Zhonghe Town in Yongning District was the center of the second-class aggregation area, and Jinchai Town in Mashan County was the center of the third-class aggregation area. Conclusion The multidrug resistant pulmonary tuberculosis epidemic in Nanning is distributed in an aggregated manner, especially in Xinyang Street, Xixiangtang District, which has the highest spatial and temporal aggregation. It is necessary to focus on and take regional prevention and control measures to control the epidemic.

Objective:To analyze the histopathological characteristics of peri-implant soft tissue in re-constructed jaws and the changes after keratinized mucosa augmentation(KM A)with free gingival graft(FGG).Methods:Twenty patients were enrolled in this study.Five patients of them,who were perio-dontal and systemic healthy and referred for crown lengthening before restoration with healthy keratinized gingiva collected were enrolled as healthy controls.15 patients of them were with fibula or iliac bone flaps jaw reconstruction(10 with fibula flap and 5 with iliac flap),who were referred to FGG and implant exposures before restoration.Soft tissue was collected before FGG in reconstructed jaws,and in 5 patients(3 with fibula flap and 2 with iliac flap)8 weeks after FGG if a second surgery was conducted.Histologi-cal analysis with hematoxylin-eosin stain and immunological analysis to interlukin-1(IL-1),interlukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were performed.Results:Thickness from the bottom of stratum basale to the top of stratum granulosum and thickness of keratinized layer in reconstructed jaws were significantly lower compared with that of natural healthy keratinized gingiva[0.27(0.20,0.30)mmvs.0.36(0.35,0.47)mm,P＜0.05;16.49(14.90,23.37)μm vs.26.37(24.12,31.53)μm,P＜0.05].In the reconstructed area,thickness from the bottom of stratum basale to the top of stra-tum granulosum increased after KMA with FGG[0.19(0.16,0.25)mm vs.0.38(0.25,0.39)mm,P=0.059]and the thickness of keratinized layer significantly increased after KMA with FGG[16.42(14.16,22.35)μm vs.28.57(27.16,29.14)μm,P＜0.05],which was similar to that in the con-trol group.Furthermore,the number of positive cells of IL-1,IL-6 and TNF-α significantly increased after KMA[0.67(0.17,8.93)vs.11.00(9.16,18.00);13.00(8.50,14.14)vs.21.89(15.00,28.12);0.22(0.04,0.63)vs.2.83(1.68,5.00),respectively,P＜0.05]as well as the average optical density value[0.15(0.14,0.17)vs.0.18(0.17,0.21);0.28(0.26,0.33)vs.0.36(0.33,0.37);0.23(0.22,0.29)vs.0.30(0.28,0.42),respectively,P＜0.05],which was similar to that in the healthy keratinized gingiva.Conclusion:The lack of rete pegs and inflammato-ry factors were common in soft tissue with jaw reconstruction.FGG can improve the quality of the epithe-lium and may improve the stability of the mucosa around implants.

Objective To explore the value of cytokines,procalcitonin(PCT)and neutrophil-lymphocyte ratio(NLR)in the early diagnosis and prognosis evaluation in the patients with sepsis.Methods 98 patients with sepsis admitted to the First Affiliated Hospital of Air Force Medical University from January 2020 to January 2023 were selected as research objects,including 82 patients in the sepsis non-shock group and 16 patients in the sepsis shock group.According to the death within 28 days,the patients was divided into survival group(n=82)and death group(n=16).Meantime,95 cases of non-septic infection group were included as control.The expression of interleukin(IL)-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,tumor necrosis factor(TNF)-α,interferon(IFN)-γ,IFN-α,PCT,and NLR were detected within 24h after admission,and their relationship with sepsis was analyzed by ROC curve.Results ①The IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,TNF-α,IFN-α,NLR and PCT in patients with sepsis were significantly higher than those in healthy subjects(Z=0.43～30.54,all P＜0.05)except IL-1β,IFN-γ.Further analysis of ROC showed that IL-8,NLR,PCT and IL-17 had strong predictive ability,with area under curve(AUC)of 0.78(95%CI:0.71～0.84),0.81(95%CI:0.75～0.87),0.83(95%CI:0.78～0.88),0.86(95%CI:0.81～0.92),respectively.Combined detection of the four indicators can effectively improve the diagnostic efficiency of sepsis,with the AUC of 0.90(95%CI:0.85～0.93).② There were no significant differences in cytokines,PCT and NLR concentration between positive and negative blood culture groups(P＞0.05),suggesting that these indexes were not affected by blood culture detection results.③Among the patients in the shock group,IL-6[122.10(10.77～10 000.00)ng/L]was significantly higher than that in non-shock group[25.56(1.02～9 096.74)ng/L],the difference was statistically significant(Z=74.55,P=0.01),with the AUC of 0.73(95%CI:0.59～0.87).The levels of IL-10[10.69(1.12～1 338.00)ng/L],IL-2[12.52(0.86～280.42)ng/L]and IL-5[9.55(0.93～259.57)ng/L]in sepsis death group were higher than those[2.55(0.34～695.13)ng/L,4.46(0.13～625.43)ng/L,2.75(0.01～117.88)ng/L]in survival group,the differences were statistically significant(Z=3.64,6.37,4.74,all P＜0.05),and the AUC were 0.69(95%CI:0.53～0.85),0.71(95%CI:0.56～0.85)and 0.72(95%CI:0.58～0.87),respectively.Conclusion The combined detection of IL-8,NLR,PCT and IL-17 is helpful for the early diagnosis of sepsis.The increase of IL-6 level can effectively predict the occurrence of septic shock,and the high expression of IL-10,IL-2 and IL-5 has a good predictive value for the death of sepsis patients.

ObjectiveTo observe the effects of Youguiwan on bone metabolism and bone morphogenetic protein-2 （BMP-2）/Smad signaling pathway in ovaries-removed rats with osteoporosis and study the mechanism of Youguiwan in the prevention and treatment of osteoporosis. MethodA postmenopausal rat model of osteoporosis was prepared by bilateral ovariectomy. The 40 female SD rats were randomly divided into five groups， including sham operation group， model group， alendronate sodium group （0.1 mg·kg^-1）， and high-dose and low-dose （5.36 and 2.68 g·kg^-1） groups of Youguiwan. The drug was given seven days after modeling， once a day for 12 weeks. After the treatment， the changes in femur tissue structure were observed by micro-CT， including bone mineral density （BMD）， bone volume/total volume （BV/TV）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， bone surface/bone volume （BS/BV）， and trabecular separation （Tb.Sp）. Ossification was observed by saffrane-solid green staining， and serum levels of bone metabolism markers， including bone alkaline phosphatase （BALP）， osteocalcin （BGP）， type Ⅰ procollagen amino terminal propeptide （PINP）， and tartrate-resistant acid phosphatase 5b （TRACP-5b）， were determined by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA expression levels of Runx2， BMP-2， and Smad1 in rat femur were detected by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the sham operation group， bone trabecula in the model group was sparse. BMD， BV/TV， Tb.N， and Tb.Th were decreased （P<0.05， P<0.01）. BS/BV （P<0.05） and Tb.Sp were increased. The content of BGP， BALP， PINP， and TRACP-5b in serum was significantly increased （P<0.01）. The mRNA and protein expressions of Runx2， BMP-2， and Smad1 in rat femur were significantly decreased （P<0.05， P<0.01）. Compared with the model group， the number of bone trabeculae in the high-dose and low-dose groups of Youguiwan was increased， and the bone microstructure was improved. BMD， BV/TV， Tb.N， and Tb.Th were increased significantly （P<0.05， P<0.01）， and BS/BV and Tb.Sp were increased. The content of bone metabolic markers decreased （P<0.05， P<0.01）. ConclusionYouguiwan has certain preventive and therapeutic effects on postmenopausal osteoporosis， and its mechanism may be related to promoting bone formation by regulating the BMP-2/Smad signaling pathway.

In recent years， the incidence of pulmonary nodules has kept rising. To give full play to the advantages of traditional Chinese medicine （TCM） in the treatment of pulmonary nodules and identify the breakthrough points of integrating TCM with Western medicine， the China Association of Chinese Medicine organized medical experts in TCM and western medicine to carry out in-depth discussion regarding this disease. The discussion encompassed the modern medical advances， TCM theories of etiology and pathogenesis， the role and advantages of TCM in the whole course management of pulmonary nodules， contents and methods of research on pulmonary nodules， and science popularization work， aiming to provide a reference for clinical practice and scientific research. After discussion， the experts concluded that the occurrence of pulmonary nodules was rooted in the deficiency of the lung and spleen and triggered by phlegm dampness， blood stasis， and Qi stagnation. TCM can treat pulmonary nodules by controlling and reducing nodules， improving physical constitution， ameliorating multi-system nodular diseases， reducing anxiety and avoiding excessive diagnosis and treatment， and serving as an alternative for patients who are unwilling or unfit for surgical treatment. At present， the optimal diagnosis and treatment strategy for pulmonary nodules has not been formed， which needs to be further studied from multiple perspectives such as clinical epidemiology， biology， and evidence-based medicine. The primary task of current research is to find out the advantages， effective prescriptions， and target populations and determine the effective outcomes of TCM in the treatment of pulmonary nodules. At the same time， basic research should be carried out to explore the etiology and biological behaviors of pulmonary nodules. The expert consensus on the diagnosis and treatment of pulmonary nodules with integrated TCM and Western medicine needs to be continuously revised to guide clinicians to conduct standardized， scientific， and accurate effective diagnosis and treatment.