ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

Objective: To evaluate the efficacy and safety of ruxolitinib combined with low-dose hormone for patients with acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: Thirty patients with aGVHD after allo-HSCT admitted to the Department of Hematology of the General Hospital of Western Theater Command from November 2021 to November 2024 were retrospectively analyzed. All patients were treated with low-dose hormone (methylprednisolone 0.3-1 mg kg -d ) combined with ruxolitinib 5-10 mg d . The efficacy and adverse reactions were observed during the follow-up period to analyze the survival outcomes of the patients. Results: A total of 30 patients with aGVHD after allo-HSCT were included in this study, consisting of 15 (50%) males and 15 (50%) females with a median age of 34 year-old (ranging from 14 to 62). Classification by disease type: there were 18 cases of acute myeloid leukemia, 4 cases of acute lymphoblastic leukemia, 4 cases of aplastic anemia, and 4 cases of myelodysplastic syndrome. Classification by aGVHD severity: there were 27 cases (90%) of Ⅱ-Ⅳ degree aGVHD and 11 cases (36.7%) of Ⅲ-Ⅳ degree aGVHD. Ruxolitinib in combination with low-dose glucocorticoid treatment yield responses in 28 (93.3%) patients, of which 27 (90%) achieved complete remission (CR), while 1 (3.3%) showed partial remission (PR). One patient (3.3%) had no response (NR), and 1 patient (3.3%) exhibited progressed disease (PD). Overall survival (OS) at 1 year of transplantation was 73.9% (95%CI 49.5% to 87.7%), progression-free survival (PFS) was 93.3% (95%CI 75.9% to 98.3%), non-relapse mortality (NRM) was 20.6% (95%CI 7.9% to 47.4%), and median survival time was 27.6 months. Conclusion: Ruxolitinib combined with low-dose hormones is safe and effective in the treatment of aGVHD after allo-HSCT.

Objectives: . The prognostic value of the pretreatment albumin-to-alkaline phosphatase ratio (AAPR) in head and neck cancer (HNC) remains uncertain. This meta-analysis aimed to evaluate the predictive role of AAPR for survival outcomes in patients with HNC. Methods: . A comprehensive search of the Cochrane Library, PubMed, and Embase databases was conducted to identify relevant studies published up to July 30, 2024. We included studies on AAPR and survival outcomes in HNC patients. Results: . Eight studies comprising 1,737 HNC patients were analyzed using random-effects models. Lower AAPR values were significantly correlated with worse overall survival (hazard ratio [HR], 2.08), progression-free survival (HR, 2.00), and disease-free survival (HR, 2.18). Sensitivity analyses confirmed the robustness of these results, with no significant publication bias detected. Conclusion . Our findings suggest that pretreatment AAPR could serve as a valuable and cost-effective prognostic indicator in HNC, potentially aiding clinicians in risk stratification and treatment decision-making. However, additional validation studies are warranted to confirm its clinical applicability.

Objective To investigate the causes and management strategies for lower limb ischemic necrosis following xenogeneic heterotopic heart transplantation from a multigene-edited pig to a rhesus monkey. Methods A xenogeneic heterotopic heart transplantation was performed on December 16, 2023, at the Institute of Experimental Animals of Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, using a quintuple-gene-edited pig as the donor and a rhesus monkey as the recipient. On postoperative day (POD) 9, the recipient monkey underwent left lower limb amputation due to ischemic necrosis. Blood samples were collected at various time points after transplantation for analysis of hematologic parameters, liver and renal function, myocardial enzymes, and coagulation profiles. Ultrasound and computed tomography (CT) were used to evaluate anastomotic patency and cardiac structure. Immunological assays, including complement-dependent cytotoxicity (CDC) and IgG/IgM antibody detection, combined with clinical observations, were employed to assess rejection type and therapeutic response. Results The recipient monkey survived for 46 days after transplantation. Echocardiography demonstrated preserved biventricular systolic function in the recipient’s native heart, with left ventricular ejection fraction (LVEF) consistently exceeding 50%. In the donor pig heart, left ventricular endocardial thickening was noted on POD 9, followed by right ventricular endocardial thickening on POD 24, while LVEF remained around 35%. No hyperacute or acute rejection was detected immunologically. CDC positivity ranged between 3.4% and 5.1%, with IgG/IgM antibody binding trends consistent with CDC results. Following amputation, the recipient exhibited elevated inflammatory markers, coagulopathy, and reactive thrombocytosis, which later normalized. Immunohistochemical staining of the necrotic limb revealed arterial and venous thrombosis; however, no T-cell or B-cell infiltration was observed in vascular structures, thrombi, nerves, muscles, fascia, or skin tissues, with CD3 and CD20 staining both negative. Conclusion Limb ischemia after xenogeneic heart transplantation may be associated with lower extremity vascular thrombosis triggered by local trauma in the context of transplantation-induced inflammatory activation and coagulation dysfunction. While no clear lymphocyte-mediated rejection was observed, further studies are needed to explore the potential role of non-lymphocyte-mediated immune mechanisms.

ObjectiveTo explore the effects of electroacupuncture at governor vessel or Jiaji acupoints on the activation of lumbar spinal cord neurons and the recovery of motor function in paralyzed hind limbs in rats with complete transection of spinal cord injury. MethodsTwenty adult female SD rats were selected. Among them， 15 rats underwent the complete transection of the 10th thoracic spinal cord and were randomly divided into the Control group， the electroacupuncture at governor vessel acupoints （GV） group and the electroacupuncture at Jiaji acupoints （JJ） group， with 5 rats in each group. The other 5 rats served as the Normal group. Electroacupuncture treatment was initiated 3 days after surgery in the GV and JJ groups and performed every other day. Basso-Beatti-Bresnahan （BBB） behavioral score was evaluated weekly. Eight weeks after the operation， cortical motor evoked potential （CMEP） detection was performed， followed by perfusion fixation， tissue harvest， frozen sectioning， and immunofluorescence staining experiments. Results① Both electroacupuncture treatments activated the expression of the excitatory marker c-Fos in L1 spinal cord neurons. The c-Fos⁺ activated neurons were mainly distributed in lamina Ⅳ， Ⅴ and Ⅶ of the L1 spinal cord， while no c-Fos⁺ neuron was observed in the Normal group and the Control group. The percentages of c-Fos⁺ neurons activated by electroacupuncture in the GV group and the JJ group were （10.23±0.15） % and （10.77±1.12） %， respectively， and there was no significant difference between the two electroacupuncture groups. ② vGluT1⁺ proprioceptive afferent nerve fibers were mainly distributed in the dorsal horn and lamina Ⅶ of intermediate zone of L1 spinal cord gray matter. In the dorsal horn of L1 spinal cord， the relative fluorescence intensity of vGluT1⁺ nerve fibers in the GV group and the JJ group were greater than that in the Control group， and that in the JJ group was greater than that in the GV group. In the lamina Ⅶ in intermediate zone of L1 spinal cord， where the central pattern generator （CPG） for hindlimb rhythmic movement is located， the relative fluorescence intensity of vGluT1⁺ nerve fibers in the GV group and the JJ group were significantly higher than that in the Control group， and the JJ group was also higher than the GV group； The lamina Ⅶ neurons innervated by vGluT1⁺ axon terminals were activated to express the excitatory marker c-Fos in the GV group and the JJ group. ③ The immunofluorescence staining results of acetylcholinesterase （ChAT） in the ventral horn of L4 spinal cord of each group showed that the number of surviving ChAT⁺ motor neurons in the GV group （19.1±3.4， P=0.000 2） and JJ group （19.0±2.9， P=0.000 2） were higher than that in the Control group （9.9±1.9）. ④ The quantitative analysis of the vGluT2⁺ excitatory axonal terminals， vGAT⁺ inhibitory axonal terminals， and the ratio of vGluT2⁺/vGAT⁺ axonal terminals ［reflecting the excitation/inhibition （E/I） balance of motor neurons］ innervating ChAT⁺ motor neurons in the ventral horn of the L4 spinal cord revealed that all three indices were significantly higher in the GV and JJ groups than in the Control group. Moreover， the E/I ratio of motor neurons in the two electroacupuncture groups gradually reached the same level as normal and showed no statistical difference from the Normal group. ⑤ The electrophysiological and behavioral detection results showed that compared with the Control group， the latencies of CMEP in the GV group and the JJ group shortened， while their amplitudes increased. The BBB score of paralyzed hindlimbs showed that after 8 weeks of treatment， the scores of both the GV group （4.25±1.26， P=0.023 4） and the JJ group （4.75±0.96， P=0.000 2） were higher than those of the Control group （1.80±1.30）， but there was no statistical difference between the two electroacupuncture groups. ConclusionElectroacupuncture at GV or JJ acupoints may promote the afferent of vGluT1⁺ proprioceptive nerve fibers into the spinal cord， indicating the possibility to activate CPG neurons in lamina Ⅶ of L1 spinal cord； facilitate the survival of motor neurons in the L4 spinal cord and restore their excitatory/inhibitory balance， improving the motor function of the paralyzed hindlimbs in rats.

ObjectiveTo observe the clinical efficacy of Shentong Zhuyutang combined with Dilongtang in the treatment of lumbar disc herniation （LDH） with Qi stagnation and blood stasis syndrome， and its effect on nucleus pulposus reabsorption and immune-inflammatory factors， exploring its therapeutic mechanism from the perspective of reabsorption. MethodsA total of 120 patients with LDH from the Fourth Clinical Medical College of Guangzhou University of Chinese Medicine， treated between June 2020 and January 2023， were randomly divided into the control group （52 cases， with 8 dropouts） and the observation group （49 cases， with 11 dropouts） according to a random number table. The control group received routine treatment， while the observation group was treated with Shentong Zhuyutang combined with Dilongtang in addition to routine treatment. Visual Analogue Scale （VAS）， Oswestry Disability Index （ODI）， Japanese Orthopaedic Association （JOA） score， and traditional Chinese medicine （TCM） syndrome score were measured before treatment and after 3 courses of treatment. Venous blood samples were collected for the determination of serological indexes. MR examination was performed during the 6-month follow-up to calculate the absorption rate. ResultsAfter treatment， both groups showed significant reductions in VAS， ODI， TCM syndrome score， serum tumor necrosis factor （TNF）-α， matrix metalloproteinase （MMP）-9， and vascular endothelial growth factor （VEGF） levels， and a significant increase in JOA score compared with pre-treatment values （P<0.05）. Compared with the control group， the observation group showed significantly lower VAS， ODI， TCM syndrome score， serum TNF-α， MMP-9， and VEGF levels， and a significantly higher JOA score （P<0.05）. The proportion of nucleus pulposus reabsorption in the observation group was 57.14% （28/49）， significantly higher than 21.15% （11/52） in the control group （χ²=6.161， P<0.05）. ConclusionShentong Zhuyutang combined with Dilongtang can effectively relieve pain， improve lumbar function， and alleviate TCM clinical symptoms in LDH patients with Qi stagnation and blood stasis syndrome. Imaging findings suggest that the treatment promotes the reabsorption of nucleus pulposus protrusion， while laboratory testing shows reduced serum levels of TNF-α， MMP-9， and VEGF， which contribute to the rehabilitation of patients.

Objective:To compare the diagnostic value of contrast-enhanced ultrasound(CEUS)combined with conventional ultrasound and magnetic resonance imaging(MRI)in distinguishing benign and malignant non-mass lesions of breast.Methods:A retrospective analysis was conducted on 131 patients with non-mass lesions of breast(76 benign cases,55 malignant cases),who were pathologically confirmed at The 910th Hospital of People's Liberation Army Joint Service Support Force from January 2019 to January 2024.The CEUS and MRI characteristics between benign and malignant non-mass lesions of breast were compared.Using pathological results as the gold standard,the receiver operating characteristic(ROC)curve were employed to compare the area under curve(AUC)value,sensitivity and specificity of CEUS+conventional ultrasound and MRI in diagnosing malignant non-mass lesions of breast.The diagnostic performances of them on non-mass lesions with different sizes(maximum diameter<10 mm,10-20 mm,and>20 mm)were further explored.Results:Conventional ultrasound identified 131 non-mass lesions of breast,whereas MRI identified 79 non-mass lesions(60.31%),40 cases with mass-like lesions(30.53%),and 12 normal breast(9.16%).The overall consistency between conventional ultrasound and MRI was higher(Kappa=0.603).After CEUS combined with conventional ultrasound,38 lesions that were classified as BI-RADS 4 by conventional ultrasound downgraded to BI-RADS 3.In the 131 patients,the specificity(71.1%)of CEUS combined with conventional ultrasound was significantly higher than that(55.3%)of MRI,with a statistically significant difference(x2=5.775,P=0.016).For all patients,the AUC value(0.828)of CEUS combined with conventional ultrasound in diagnosing malignant non-mass lesions of breast was higher than that(0.776)of MRI(Z=3.246,P=0.006).Among of them,the diagnostic AUC value of CEUS combined with ultrasound was 0.842 for the patients whose size of the non-mass lesions of breast was between 10-20 mm,which was significantly larger than that(0.758)of MRI(Z=4.856,P<0.001).Conclusions:The diagnostic specificity of CEUS+conventional ultrasound is higher than that of MRI for malignant non-mass lesions of breast,which has a certain of advantage.

Objective:To investigate the predictive value of 18F-FDG PET related metabolic parameters on microsatellite instability-high (MSI-H) and human epidermal growth factor receptor-2(HER2) expression in colorectal cancer (CRC). Methods:The 18F-FDG PET imaging, clinical and pathological data of 101 CRC patients (58 males, 43 females; age 68.0(58.0, 75.0) years) admitted to Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University from January 2016 to March 2024 were retrospectively collected, including 17 cases in MSI-H group, 84 cases in microsatellite stability/microsatellite instability-low (MSS/MSI-L) group, 23 cases in HER2 expression group, 34 cases in non-HER2 expression group, and 44 patients without HER2 detection. Univariate analyses (independent-sample t test, Mann-Whitney U test, χ2 test) and multivariate logistic regression analysis were used to screen out independent risk factors, and ROC curve was used to evaluate the predictive efficacy. Bootstrap method was used to verify the model internally. Results:There were significant differences in metabolic tumor volume (MTV) 80% between MSI-H group and MSS/MSI-L group (2.1(1.6, 4.0) vs 1.4(1.0, 2.7) cm 3;Z=-2.10, P=0.036), and total lesion glycolysis (TLG) 80% and carcinoembryonic antigen (CEA) were significantly different between those 2 groups ( Z=-2.27, χ2=6.40, both P<0.05). There were significant differences in TLG 80% (29.0(16.1, 41.0) vs 14.3(9.4, 22.9) g; Z=-2.80, P=0.005) between HER2 expression group and non-HER2 expression group, and significant differences were also found in MTV 80%, heterogeneity index (HI) and CV ( Z=-2.24, t values: -2.26, 2.54, all P<0.05). The independent risk factors for MSI-H were MTV 80% (odds ratio ( OR)=1.326, 95% CI: 1.015-1.733, P=0.038) and CEA ( OR=0.200, 95% CI: 0.056-0.706, P=0.012), with the AUC for the combined model of 0.730 (95% CI: 0.605-0.856), and the concordance index (C-index) of 0.716. The independent risk factors for HER2 expression were TLG 80% ( OR=1.037, 95% CI: 1.001-1.073, P=0.041) and CV ( OR=1.467, 95% CI: 1.073-2.005, P=0.016), with the AUC for the combined model of 0.775 (95% CI: 0.645-0.875), and the C-index of 0.757. Conclusions:18F-FDG PET can be used as a noninvasive tool to evaluate CRC microsatellite status and HER2 gene amplification. MTV 80% and CEA are independent risk factors for MSI-H; TLG 80% and CV are independent risk factors for HER2 expression.

Objective:To evaluate the clinical prognostic value of intratumoral metabolic heterogeneity parameters of baseline 18F-FDG PET/CT in primary cutaneous malignant melanoma (CMM). Methods:From October 2015 to July 2023, the clinical data of 35 patients (24 males, 11 females, age (66.6±13.1) years) diagnosed with primary CMM who underwent baseline 18F-FDG PET/CT in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School were retrospectively analyzed. Conventional metabolic parameters (SUV max, metabolic tumor volume (MTV), total lesion glycolysis (TLG)) and PET intratumoral metabolic heterogeneity parameters (area under the cumulative SUV histograms curve (AUC-CSH), linear regression slope, SUV max/ SUV mean) were assessed. Using thresholds of 30%, 40%, 50%, 60%, 70%, and 80% SUV max or thresholds of 40%, 60%, and 80% SUV max to delineate MTV, linear regression was performed, with the slopes being heterogeneity index-1 (HI-1) and heterogeneity index-2 (HI-2), respectively. Using SUV thresholds of 2.5, and 40%, 50%, 60%, and 70%SUV max to calculate AUC-CSH and SUV max/SUV mean. Kaplan-Meier survival curves and Cox proportional hazards models were used to analyze the prognostic value of primary lesion PET metabolic parameters on overall survival (OS) and progression-free survival (PFS). Results:The median follow-up time of 35 patients was 20 months, with 25 patients (71%) experiencing disease progression and 16 patients (46%) deceased. Multivariate Cox regression analysis revealed that HI-1, HI-2, MTV, SUV max/SUV mean2.5, and SUV max were the independent prognostic factors for PFS (hazard rate ( HR) (95% CI): 0.32(0.13-0.82), 0.32(0.13-0.82), 3.86(1.34-11.12), 4.61(1.33-16.02), 4.06(1.55-10.61), all P<0.05), whereas SUV max and SUV max/SUV mean2.5 were the independent prognostic factors for OS ( HR: 8.04(1.96-32.87), 2.87(1.09-7.51), P values: 0.004, 0.032). Conclusion:HI-1, HI-2, and SUV max/SUV mean2.5 have prognostic value for CMM, while the value of AUC-CSH heterogeneity parameters are not significant.

Objective:To establish a combined model of tumor heterogeneity metabolic parameters using 18F-FDG PET/CT and explore its value in differentiating benign from malignant pulmonary lesions. Methods:A total of 251 patients (157 males, 94 females; age 15-88 years) who were diagnosed with malignant lung lesions by 18F-FDG PET/CT and with definitive pathological results at Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from February 2017 to February 2024 were retrospectively enrolled. Analysis was conducted on clinical data, traditional parameters (SUV max, metabolic tumor volume (MTV), total lesion glycolysis (TLG)) of primary lesions on 18F-FDG PET/CT, and intra-tumoral metabolic heterogeneity index (HI; such as cumulative SUV volume histogram AUC (AUC-CSH), linear regression slope, CV). AUC-CSH and CV were calculated using SUV thresholds of 2.5 and 40%SUV max. Logistic univariate and multivariate regression analyses were used to extract independent predictors in clinical features and PET/CT parameters for the differential diagnosis of pulmonary lesions. A multi-parameter combined model was established through logistic regression and validated for diagnostic efficacy using ROC curve analysis. Results:Among 251 patients, 101 were benign and 150 were malignant. In univariate analysis, gender, age, tumor markers, spiculation sign, lobulation sign, vessel convergence sign, air bronchogram, long diameter, short diameter, SUV max, AUC-CSH 2.5, AUC-CSH 40%, CV2.5, and CV40% were predictive factors for the diagnosis of benign and malignant tumors (odds ratio ( OR): 0.57-17.39, all P<0.05). In multivariate analysis, gender, age, tumor markers, lobulation sign, vessel convergence sign, SUV max, AUC-CSH 40%, and CV40% were independent predictors for the diagnosis of benign and malignant tumors ( OR: 2.30-13.18, all P<0.05). The AUC, sensitivity, specificity, and accuracy of the multi-parameter combined model established with the above independent predictors were 0.89, 77.33%(116/150), 84.16%(85/101), 80.08%(201/251), respectively. Conclusion:18F-FDG PET/CT multi-parameter combined model has high value in the differentiation of benign and malignant pulmonary lesions.