Objective To evaluate the clinical utility of metagenomic next-generation sequencing(mNGS) for etiologic diagnosis in severe pneumonia and to compare its performance with conventional microbiological testing and serological assays. Methods We retrospectively included 127 patients with severe pneumonia admitted to the intensive care unit of the Department of Pulmonary and Critical Care Medicine Ⅱ, The Second Hospital of Hebei Medical University from February2021 to October 2022; all underwent m NGS testing. Bronchoalveolar lavage fluid(BALF) was collected via flexible bronchoscopy. Samples underwent m NGS, conventional microbiology(smear, culture, and acid-fast staining), and serological testing(IgM panel for nine respiratory pathogens and PCR-based nucleic acid detection). Pathogen positivity rates and inter-method agreement were compared to assess the accuracy and timeliness of m NGS testing. Results The overall positivity rate of m NGS testing was 89. 76%, significantly higher than that of conventional microbiology(63. 78%) and serology testing(51.18%)(all P <0.05). For bacterial detection, the m NGS positivity rate(71.65%) exceeded those of conventional microbiology(55.12%,P = 0.006) and serology testing(10.24%, P <0.001). For fungal detection, m NGS likewise outperformed conventional microbiology(62.99% vs. 21.26%) and serology testing(11.81%), with all pairwise differences achieving statistical significance(all P <0.001). In contrast, serology testing was superior to m NGS for detecting atypical pathogens(9.45% vs.2.36%,P = 0.017). Agreement between m NGS and either conventional microbiology or serology testing was poor(Kappa <0.4). For mixed infections, m NGS yielded a significantly higher positivity rate(65.35%) than conventional microbiology(11.81%) and serology testing(17.32%), with all pairwise differences achieving statistical significance(all P <0.001). Conclusion m NGS demonstrates a high positivity rate and offers broad, rapid detection advantages for etiologic diagnosis in severe pneumonia, with particularly strong performance for bacterial and fungal pathogens and for identifying mixed infections.

Objective To investigate the expression levels of death-associated protein 3(DAP3) and programmed cell death molecule 10(PDCD10) in hepatocellular carcinoma(HCC) tissues, analyze their correlation with invasion and metastasis indicators, and evaluate their predictive value for the prognosis of HCC patients. Methods A total of 148 HCC tissue specimens surgically resected from the Department of Oncology, Yibin First People ' s Hospital from February 2020 to February 2022 were selected. Immunohistochemistry was performed to detect the protein expression levels of DAP3 and PDCD10. Quantitative real-time PCR was used to detect the m RNA expression of DAP3, PDCD10, and invasion and metastasis-related genes Snail, N-cadherin(N-cad), and E-cadherin(E-cad). The Kaplan-Meier method and Cox proportional hazards regression model were used to identify independent factors affecting the prognosis of HCC patients. Results The positive rate of DAP3 in cancer tissues [70.27%(104/148) vs. 6.76%(10/148)]was significantly higher than that in adjacent tissues;the positive rate of PDCD10 in cancer tissues [74.32%(110/148) vs. 5.41%(8/148)] was also significantly higher than that in adjacent tissues(χ2= 126.058, 146.619, both P <0.001). The m RNA expression levels of DAP3, PDCD10, Snail, and N-cad were significantly increased in HCC cancer tissues, while E-cad m RNA expression was significantly decreased(t = 52.620,48.361, 55.643, 39.695, 28.074, all P <0.001). The m RNA expression of DAP3 and PDCD10 in HCC tissues was positively correlated with Snail and N-cad m RNA(r = 0.668, 0.710; 0.654, 0.698, all P <0.001), and negatively correlated with E-cad m RNA expression(r =-0.654,-0.725, all P <0.001). The protein expression of DAP3 and PDCD10 in HCC tissues from patients with CNLC stage Ⅱ–Ⅲ was significantly higher than that in patients with CNLC stage Ⅰ(χ2= 13.954/<0.001, 4.913/0.027). The 3-year overall survival rate of the DAP3-positive group was 34.62%(36/104), which was significantly lower than that of the DAP3-negative group(84.09%, 37/44)(Log-rank χ2= 18.270, P <0.001). The 3-year overall survival rate of the PDCD10-positive group was 37. 27%(41/110), which was significantly lower than that of the PDCD10-negative group(84.21%, 32/38)(Log-rank χ2= 11.000,P = 0.001). CNLC stage Ⅱ–Ⅲ, DAP3 positivity, and PDCD10 positivity were independent risk factors affecting the prognosis of HCC patients [HR(95%CI) = 1.603(1.167-2.202), 1.840(1.319-2.568), 1.775(1.275-2.473)]. Conclusion The elevated expression of DAP3 and PDCD10 in HCC is closely associated with tumor invasion and metastasis, and these proteins may serve as potential biomarkers for evaluating the prognosis of HCC patients.

Objective To investigate the predictive value of serum macrophage colony-stimulating factor(M-CSF)and intercellular adhesion molecule-1(ICAM-1) levels for postoperative adhesive intestinal obstruction in children. Methods A total of 152 pediatric patients with intestinal adhesions after abdominal surgery who were admitted to the Department of General Surgery of the First Hospital of Hebei Medical University from January 2023 to June 2024 were enrolled. Based on imaging findings from upright abdominal radiographs, abdominal ultrasonography, and computed tomography(CT), patients were divided into an intestinal obstruction subgroup(n = 50) and a non-obstruction subgroup(n = 102). In addition, 50 age-and sex-matched healthy children who underwent physical examinations during the same period were included as the healthy control group. Fasting venous blood samples were collected from all participants, and serum levels of M-CSF and ICAM-1 were measured using enzyme-linked immunosorbent assay(ELISA). Pearson correlation analysis was performed to evaluate the relationships between serum M-CSF and ICAM-1 levels and inflammatory indicators, including white blood cell(WBC) count,C-reactive protein(CRP), and procalcitonin(PCT). Multivariate logistic regression analysis was used to identify independent risk factors for postoperative adhesive intestinal obstruction. Receiver operating characteristic(ROC) curves were generated to assess the predictive performance of serum M-CSF and ICAM-1 levels. Results Serum M-CSF and ICAM-1 levels were significantly higher in the adhesive intestinal obstruction group than in the healthy control group(M-CSF:t = 11.152,P <0.001;ICAM-1: t = 8.701,P <0.001). The incidence of intestinal obstruction among the 152 patients with adhesions was 32.89%(50/152). The intestinal obstruction subgroup showed significantly higher WBC, CRP, and PCT levels compared with the non-obstruction subgroup(WBC:t = 13.816,P <0.001; CRP:t = 34.123,P <0.001; PCT:t = 19.216,P <0.001). Pearson correlation analysis indicated that serum M-CSF levels were positively correlated with WBC(r = 0.683,P = 0.013), CRP(r = 0.701,P = 0.008),and PCT(r = 0.782,P = 0.027), while serum ICAM-1 levels were positively correlated with WBC(r = 0.659, P = 0.024), CRP(r = 0.712,P = 0.011), and PCT(r = 0.747,P = 0.003). Multivariate logistic regression analysis revealed that elevated WBC, CRP,and PCT levels were independent risk factors for postoperative adhesive intestinal obstruction [WBC: OR = 1.317, 95%CI(1.233-3.989); CRP: OR = 2.429, 95%CI(1.506-4.038); PCT:OR = 1.103, 95%CI(0.738-2.899)]. The area under the ROC curve(AUC) for M-CSF, ICAM-1, and their combined detection in predicting adhesive intestinal obstruction was 0.694, 0.752,and 0.820, respectively. The AUC of the combined detection was higher than that of either marker alone. Conclusion Serum M-CSF and ICAM-1 levels are elevated in children with postoperative adhesive intestinal obstruction and are associated with disease severity. Combined detection of these two biomarkers provides a higher predictive value for the occurrence of postoperative adhesive intestinal obstruction than individual testing.

Objective To investigate the correlation and clinical value of mi R-34a and PSPC1 expression in cancerous tissues with clinicopathological features of oral squamous cell carcinoma(OSCC). Methods A total of 101 OSCC patients(OSCC group) and 68 patients with benign oral diseases(control group) treated at Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2020 to January 2023 were selected as the study subjects.The expression of mi R-34a and PSPC1 was detected. Spearman rank correlation or Pearson correlation analysis was used to analyze their correlation with clinicopathological features. Receiver operating characteristic(ROC) curves and De Long' s method were used to analyze sensitivity, specificity, and AUC. Cox regression analysis was used to identify adverse prognostic risk factors. Kaplan-Meier curves were used for survival analysis. Results Mi R-34a expression in OSCC tissue was lower than that in adjacent normal tissue and the control group, while PSPC1 expression was higher than that in adjacent normal tissue and the control group(F/P = 401.394/<0.001, 215.425/<0.001). Patients with t TMB ≥20 mut/Mb, ct DNA positive, CTC positive, moderately or poorly differentiated tumors, and TNM stage Ⅲ-Ⅳ showed lower mi R-34a expression than patients with t TMB < 20 mut/Mb, ct DNA negative, CTC negative, well-differentiated tumors, and TNM stage Ⅰ-Ⅱ(t/P = 2.919/0.004,3.881/<0.001, 5.937/<0.001, 4.223/<0.001, 8.371/<0.001). PSPC1 expression was higher in patients with t TMB ≥20 mut/Mb,ct DNA positive, CTC positive, moderately or poorly differentiated tumors, and TNM stage Ⅲ-Ⅳ(t/P = 4.226/<0.001, 5.177/<0.001, 6.923/<0.001, 5.024/<0.001, 6.127/<0.001). In the OSCC group, mi R-34a expression was negatively correlated with t TMB, ct DNA, CTCs, pathological grade, and TNM stage(rs/P =-0. 659/0. 021,-0. 617/0. 035,-0. 645/0. 018,-0. 629/0.007,-0.627/0.029). PSPC1 expression was positively correlated with t TMB, ct DNA, CTCs, pathological grade, and TNM stage(rs/P = 0.605/0.011, 0.633/0.003, 0.645/0.031, 0.619/0.029, 0.644/0.025). The combination of mi R-34a and PSPC1 was significantly more effective than either marker alone in predicting poor prognosis in patients with oral squamous cell carcinoma(Z/P = 7.012/<0.05, 6.848/<0.05). Mi R-34a ≤0.79, PSPC1 ≥0.86, t TMB ≥20 mut/Mb, ct DNA positive, CTC positive, moderately to poorly differentiated pathological grade, and TNM stage Ⅲ-Ⅳ were risk factors for poor prognosis of oral squamous cell carcinoma[HR(95%CI) = 4.162(1.322-7.002), 3.357(1.276-5.438), 2.517(1.089-3.945), 2.447(1.013-3.882),2.748(1.127-4.370), 1.844(1.056-2.632), 2.106(1.101-3.112)]. OSCC patients with mi R-34a ≤0.79 and PSPC1 ≥0.86 had a significantly lower median survival than those with mi R-34a >0.79 or PSPC1 <0.86(Log Rank χ2= 9.421,P <0.001). Conclusion The expression of mi R-34a and PSPC1 in cancerous tissue is significantly associated with disease severity, poor prognosis, and overall survival in OSCC. Combined detection of both markers has greater clinical value in OSCC.

Objective To study the expression of transmembrane p24 trafficking protein 2(TMED2) and sialic acidbinding immunoglobulin-type lectins 15(Siglec15) in cutaneous malignant melanoma(CMM) and their clinicopathological features, immune cell infiltration, and prognostic significance. Methods A total of 109 patients with CMM admitted to the Department of Dermatology, the Fifth Medical Center of Chinese PLA General Hospital from April 2019 to April 2022 were selected as the research subjects. The protein and m RNA expressions of TMED2 and Siglec15 in CMM tissues were detected by immunohistochemical staining and quantitative real-time PCR(qPCR). R language was used to analyze the relationship between TMED2, Siglec15 expression and immune cell infiltration in the TCGA database. Kaplan-Meier survival analysis and Cox multivariate regression analysis were used to evaluate the effect of TMED2 and Siglec15 protein expression on the prognosis of CMM patients. Results The positive rates of TMED2 and Siglec15 in cancer tissues were 67.89%(74/109) and64.22%(70/109), respectively, which were significantly higher than those in adjacent tissues [7.34%(8/109) and 9.17%(10/109)](χ2/P = 85.151/<0.001, 71.087/<0.001). The m RNA expression levels of TMED2 and Siglec15 in CMM cancer tissues were 2.52 ± 0.54 and 3.03 ± 0.46, respectively, which were significantly higher than those in adjacent tissues(0.93±0.25 and1.05±0.28)(t/P = 27.896/<0.001, 38.387/<0.001). Analysis of TCGA database showed that TMED2 expression in CMM tissues was positively correlated with Treg cells, macrophages, and neutrophils(r/P = 0.310/<0.001, 0.254/<0.001, 0.226/<0.001), and negatively correlated with CD56+NK cells, CD8+T cells, and NK cells(r/P =-0.407/<0.001,-0.344/<0.001,-0.298/<0.001).Siglec15 expression was positively correlated with Treg cells, macrophages, and follicular helper T cells(TFH)(r/P = 0.433/<0.001, 0.488/<0.001, 0.416/<0.001). The positive rates of TMED2 and Siglec15 in CMM tissues were significantly higher in patients with clinical stage ⅢA – B and lymph node metastasis(χ2/P = 11.957/0.001, 10.366/0.001; 12.789/<0.001, 17.217/<0.001). The 3-year overall survival rate of the TMED2-positive group was 68.92%(51/74), which was significantly lower than that of the negative group(85.71%, 30/35)(Log-rank χ2= 3.320,P <0.001). The 3-year overall survival rate of the Siglec15-positive group was 67.14%(47/70), which was significantly lower than that of the negative group(87.18%, 34/39)(Log-rankχ2= 5.580,P <0.001). Clinical stage ⅢA – B, lymph node metastasis, TMED2 positivity, and Siglec15 positivity were independent risk factors affecting the prognosis of CMM patients [HR(95%CI) = 1.665(1.145-2.421), 1.383(1.101-1.736), 1.508(1.192-1.908), 1.391(1.139-1.699)]. Conclusion The elevated expression of TMED2 and Siglec15 in CMM is associated with tumor immune infiltration and adverse clinicopathological features, and they represent novel biomarkers for evaluating the prognosis of CMM.