Our previous research demonstrated that the Wenxia Changfu Formula (WCF), as a neoadjuvant therapy, inhibits M2 macrophage infiltration in the tumor microenvironment and prevents lung cancer metastasis. Given tumor-associated macrophages (TAMs) in epithelial-mesenchymal transition (EMT), this study investigated whether WCF impedes lung cancer metastasis by attenuating TAM-induced EMT in non-small cell lung cancer (NSCLC) cells. Utilizing a co-culture model treated with or without WCF, we observed that WCF downregulated cluster of differentiation 163 (CD163) expression in macrophages, reduced CCL18 levels in the conditioned medium, and inhibited the growth, invasion, and EMT of NSCLC cells induced by macrophage co-culture. Manipulation of CCL18 levels and Src overexpression in NSCLC cells revealed that WCF's effects are mediated through CCL18 and Src signaling. In vivo, WCF inhibited recombinant CCL18 (rCCL18)-induced tumor metastasis in nude mice by blocking Src signaling. These findings indicate that WCF inhibits NSCLC metastasis by impeding TAM-induced EMT via antagonistic modulation of CCL18, providing evidence for its potential development and clinical application in NSCLC patients.
Epithelial-Mesenchymal Transition/drug effects* ; Carcinoma, Non-Small-Cell Lung/metabolism* ; Humans ; Animals ; Lung Neoplasms/metabolism* ; Chemokines, CC/antagonists & inhibitors* ; Mice ; Mice, Nude ; Drugs, Chinese Herbal/administration & dosage* ; Cell Line, Tumor ; Neoplasm Metastasis ; Tumor-Associated Macrophages/drug effects* ; Mice, Inbred BALB C ; Signal Transduction/drug effects*

Objective:To validate and optimize the interpretation methods and cut-off values of prolonged activated partial thromboplastin time (APTT) mixing test, in order to elevate the clinical applicational value in differentiating single factor deficiency, positive lupus anticoagulant (LA) and FⅧ inhibitors.Methods:This cross-sectional retrospective study involved 130 cases with single APTT prolongation for unknown reasons who underwent mixing tests in the First Hospital of Jilin University from January 1st, 2022 to December 31st, 2023, including 84 males and 46 females. Rosner index (RI), Percentage correction ("Chang") method, Normal reference interval (NRI) method, differential analysis, post-incubation prolongation time and percentage (including mixed immediately and ) were respectively calculated and analyzed for both immediate mixing and 2-hour post-mixing incubation. According to clinical diagnosis, patients were divided into single factor deficiency (51 cases), positive lupus anticoagulant (LA) (56 cases), and positive factor Ⅷ (FⅧ) inhibitor (23 cases). Receiver operation characteristic (ROC) curve analysis was used to determine the cut-off values for APTT mixing test methods in patients with different reasons for prolonged APTT, which were evaluted and adjusted by AUC, sensitivity, and specificity, and compared with the methods currently used for interpretations.Results:For the single factor deficiency cases, the AUCs of immediate RI index <7.5% and incubated RI index <13.9% were 0.87 and 0.90, respectively, with sensitivities at 87.8% and 83.8%, and specificities at 72.0% and 80.0%, whose combination had sensitivity and specificity at 87.3%, and 94.0%, respectively; the judging accuracy increased from 84.6% (110/130) in currently used rule to 93.1% (121/130). For the positive lupus anticoagulant (LA) cases, the combination of immediate RI index >10.3% and prolonged post-incubation clotting time percentage <9.1% showed 88.5% sensitivity and 99.8% specificity; the judging accuracy improved from 80.0% (104/130) in currently used rule to 86.2% (112/130). For positive FⅧ inhibitor cases, the combination of incubated RI index >17.0% and prolonged post-incubation clotting time percentage >9.1% had 99.8% sensitivity and 100.0% specificity; the judging accuracy increased from 89.2% (116/130) in currently used rule to 94.6% (123/130).Conclusion:This study validated and optimized the interpretation methods and cut-off values for the prolonged APTT mixing test in differentiating single factor deficiencies, prositive FⅧ inhibitors and LA positive cases, significantly improving the judging sensitivity and specificity.

Objective:To validate and optimize the interpretation methods and cut-off values of prolonged activated partial thromboplastin time (APTT) mixing test, in order to elevate the clinical applicational value in differentiating single factor deficiency, positive lupus anticoagulant (LA) and FⅧ inhibitors.Methods:This cross-sectional retrospective study involved 130 cases with single APTT prolongation for unknown reasons who underwent mixing tests in the First Hospital of Jilin University from January 1st, 2022 to December 31st, 2023, including 84 males and 46 females. Rosner index (RI), Percentage correction ("Chang") method, Normal reference interval (NRI) method, differential analysis, post-incubation prolongation time and percentage (including mixed immediately and ) were respectively calculated and analyzed for both immediate mixing and 2-hour post-mixing incubation. According to clinical diagnosis, patients were divided into single factor deficiency (51 cases), positive lupus anticoagulant (LA) (56 cases), and positive factor Ⅷ (FⅧ) inhibitor (23 cases). Receiver operation characteristic (ROC) curve analysis was used to determine the cut-off values for APTT mixing test methods in patients with different reasons for prolonged APTT, which were evaluted and adjusted by AUC, sensitivity, and specificity, and compared with the methods currently used for interpretations.Results:For the single factor deficiency cases, the AUCs of immediate RI index <7.5% and incubated RI index <13.9% were 0.87 and 0.90, respectively, with sensitivities at 87.8% and 83.8%, and specificities at 72.0% and 80.0%, whose combination had sensitivity and specificity at 87.3%, and 94.0%, respectively; the judging accuracy increased from 84.6% (110/130) in currently used rule to 93.1% (121/130). For the positive lupus anticoagulant (LA) cases, the combination of immediate RI index >10.3% and prolonged post-incubation clotting time percentage <9.1% showed 88.5% sensitivity and 99.8% specificity; the judging accuracy improved from 80.0% (104/130) in currently used rule to 86.2% (112/130). For positive FⅧ inhibitor cases, the combination of incubated RI index >17.0% and prolonged post-incubation clotting time percentage >9.1% had 99.8% sensitivity and 100.0% specificity; the judging accuracy increased from 89.2% (116/130) in currently used rule to 94.6% (123/130).Conclusion:This study validated and optimized the interpretation methods and cut-off values for the prolonged APTT mixing test in differentiating single factor deficiencies, prositive FⅧ inhibitors and LA positive cases, significantly improving the judging sensitivity and specificity.

Inflammatory bowel disease (IBD) is a chronic gastrointestinal disorder resulting from genetic susceptibility, abnormal immune function in the intestinal mucosa, disruptions of gut microbiota, and other factors. With the population aging, the numbers of elderly patients with IBD and those with Alzheimer diseases (AD) are on the rise. Studies have demonstrated that there is a significant association between IBD and AD, as both conditions share similar pathophysiological mechanisms: immune imbalance, chronic inflammation and gut microbiota disruptions. This article reviews the comorbidity between IBD and AD and the common pathological mechanisms of these two conditions to provide reference for clinical management of these patients.

Objective:To establish a set of artificial intelligence (AI) verification rules for blood routine analysis.Methods:Blood routine analysis data of 18 474 hospitalized patients from the First Hospital of Jilin University during August 1st to 31st, 2019, were collected as training group for establishment of the AI verification rules,and the corresponding patient age, microscopic examination results, and clinical diagnosis information were collected. 92 laboratory parameters, including blood analysis report parameters, research parameters and alarm information, were used as candidate conditions for AI audit rules; manual verification combining microscopy was considered as standard, marked whether it was passed or blocked. Using decision tree algorithm, AI audit rules are initially established through high-intensity, multi-round and five-fold cross-validation and AI verification rules were optimized by setting important mandatory cases. The performance of AI verification rules was evaluated by comparing the false negative rate, precision rate, recall rate, F1 score, and pass rate with that of the current autoverification rules using Chi-square test. Another cohort of blood routine analysis data of 12 475 hospitalized patients in the First Hospital of Jilin University during November 1sr to 31st, 2023, were collected as validation group for validation of AI verification rules, which underwent simulated verification via the preliminary AI rules, thus performance of AI rules were analyzed by the above indicators. Results:AI verification rules consist of 15 rules and 17 parameters and do distinguish numeric and morphological abnormalities. Compared with auto-verification rules, the true positive rate, the false positive rate, the true negative rate, the false negative rate, the pass rate, the accuracy, the precision rate, the recall rate and F1 score of AI rules in training group were 22.7%, 1.6%, 74.5%, 1.3%, 75.7%, 97.2%, 93.5%, 94.7%, 94.1, respectively.All of them were better than auto-verification rules, and the difference was statistically significant ( P<0.001), and with no important case missed. In validation group, the true positive rate, the false positive rate, the true negative rate, the false negative rate, the pass rate, the accuracy, the precision rate, the recall rate and F1 score were 19.2%, 8.2%, 70.1%, 2.5%, 72.6%, 89.2%, 70.0%, 88.3%, 78.1, respectively, Compared with the auto-verification rules, The false negative rate was lower, the false positive rate and the recall rate were slightly higher, and the difference was statistically significant ( P<0.001). Conclusion:A set of the AI verification rules are established and verified by using decision tree algorithm of machine learning, which can identify, intercept and prompt abnormal results stably, and is moresimple, highly efficient and more accurate in the report of blood analysis test results compared with auto-vefication.

Objective:Clot waveform analysis (CWA) of activated partial thromboplastin time (APTT) was used to rapidly identify patients with hemophilia A (HA) and lupus anticoagulant (LA) positive, and to explore the diagnostic efficacy of APTT-CWA in HA and LA-positive patients.Methods:From July 25, 2022, to December 28, 2023, 145 patients with APTT prolongation were admitted to the First Hospital of Jilin University. This group comprised 106 males and 39 females, with an average age of (29.3±15.8) years, among whom there were 94 clinically confirmed HA cases and 51 LA-positive cases. The retrospective analysis employed the Mann-Whitney U test to compare APTT results and CWA parameters between HA patients and LA-positive individuals.CWA parameters including the first derivative of maximum reaction velocity (|min1|), the second derivative of maximum acceleration (|min2|), and maximum deceleration (|max2|) were investigated. Patients were classified based on the duration of APTT prolongation into mild (33 s53 s) groups. A comparison of APTT seconds and CWA parameters |min1|, |min2|, and |max2| was conducted for HA and LA-positive patients between each group. Furthermore, receiver operating characteristic (ROC) curves and area under the curve (AUC) were utilized to assess the diagnostic efficacy of CWA parameters and determine cut-off values. Results:The APTT results for HA and LA-positive patients were 70.2 s and 41.5 s, respectively, exhibiting a statistically significant difference ( P<0.001). CWA parameters |min1| were 1.459 %/s vs 3.868 %/s, |min2| were 0.124 %/s2 vs 0.502 %/s2, and |max2| were 0.054 %/s2 vs 0.388 %/s2in HA and LA patients, respectively ( P<0.001). In all APTT prolongation groups, the CWA parameters |min1|, |min2|and |max2| for HA were significantly lower than those for LA-positive patients ( P<0.05). CWA graph analysis revealed that the peak heights of |min1|, |min2|, and |max2| in HA were lower compared to LA patients. In the severe extension group, HA displayed significantly decreased peak values and a flatter, incomplete curve. ROC curve analysis indicated AUC values were greater than 0.8 ( P<0.01) for |min1|, |min2|, and |max2| in all APTT prolongation groups. Cut-off values for |min1|, |min2|, and |max2| were established for each prolongation group: mild (3.205 %/s, 0.423 %/s2, and 0.300 %/s2), moderate (2.884 %/s, 0.340 %/s2, 0.231 %/s2), and severe (2.209 %/s, 0.190 %/s2, 0.095 %/s2). Conclusion:When APTT was extended within the same range, there were significant differences in CWA parameters between HA patients and LA patients, ROC curve analysis demonstrated the effectiveness of CWA parameters in distinguishing HA from LA-positive patients,APTT-CWA can serve as a low-cost, efficient, and specific method for the rapid identification of hemophilia and lupus anticoagulant-positive patients.

Objective:To evaluate the predictive value of preoperative frailty combined with nutritional status for prolonged postoperative ileus (PPOI) in the patients with gynecological malignancies.Methods:Patients undergoing elective surgery for gynecological malignancies in the Affiliated Hospital of Jiangnan University from April 2022 to February 2023 were selected. The Frail scale was used to evaluate the frailty within 24 h of admission, and the nutritional status was evaluated by the Controlling Nutritional Status score. The general characteristics of patients and occurrence of PPOI were recorded, and the risk factors for PPOI were analyzed by multivariate logistic regression. The ability of frailty, nutritional status and their combination to predict PPOI was assessed by the receiver operating characteristic curve.Results:Two hundred and fourteen patients were finally included, 52 cases developed of PPOI, and 98 cases were frail patients. Preoperative frailty combined with moderate to severe malnutrition was an independent risk factor for PPOI in the patients with gynecological malignancies ( P<0.05), and the area under the curve in predicting the occurrence of PPOI was 0.796 (95% confidence interval 0.736-0.857) in the patients with gynecological malignancies. Conclusions:Preoperative frailty combined with moderate to severe malnutrition has a higher accuracy in predicting PPOI in the patients with gynecological malignancies.

Objective:To evaluate the role of Z-DNA-binding protein 1 (ZBP1)/receptor-interacting protein kinase 1 (RIPK1) signaling pathway in lipopolysaccharide (LPS)-adenosine triphosphate (ATP)-induced pyroptosis in macrophages of mice.Methods:The RAW264.7 macrophages from mice were routinely cultured and divided into 6 groups ( n=9 each) using a random number table method: control group (group C), LPS-ATP group, LPS-ATP+ transfection negative control scRNA group (group LPS-ATP+ scRNA), LPS-ATP+ ZBP1 small interference RNA group (group LPS-ATP+ siRNA), LPS-ATP+ dimethyl sulfoxide group (group LPS-ATP+ DSMO), and LPS-ATP+ RIPK1 inhibitor nec-1 group (group LPS-ATP+ nec-1). The siRNA technique was used to inhibit the expression of ZBP1 in group LPS-ATP+ siRNA. The RIPK1 inhibitor nec-1 was given to inhibit the expression of RIPK1 protein in group LPS-ATP+ nec-1. Group C was routinely cultured. Cells were incubated with 10 μg/ml LPS for 24 h, then 5 mmol/L ATP was added, and the cells were incubated for 30 min to develop the cell pyroptosis model in the remaining 5 groups. The cell survival was detected by the CCK-8 assay. The concentrations of interleukin-1beta (IL-1β), IL-6, IL-18 and tumor necrosis factor-alpha (TNF-α) in cell supernatant were determined by enzyme-linked immunosorbent assay. The pyroptosis was determined by propidium iodide fluorescence staining. Western blot was used to detect the expression of ZBP1, RIPK1, caspase-1 and GSDMD. Results:Compared with group C, the cell survival rate was significantly decreased, the cell pyroptosis rate and concentrations of IL-1β, IL-6, IL-18 and TNF-α in the supernatant were increased, and the expression of ZBP1, RIPK1, caspase-1 and GSDMD was up-regulated in group LPS-ATP ( P<0.05). Compared with group LPS-ATP, no significant change was found in the parameters mentioned above in group LPS-ATP+ scRNA and group LPS-ATP+ DSMO ( P>0.05). Compared with group LPS-ATP+ scRNA, the cell survival rate was significantly increased, the cell pyroptosis rate and concentrations of IL-1β, IL-6, IL-18 and TNF-α in the supernatant were decreased, and the expression of ZBP1, RIPK1, caspase-1 and GSDMD was down-regulated in group LPS-ATP+ siRNA ( P<0.05). Compared with group LPS-ATP+ DMSO, the cell survival rate was significantly increased, the cell pyroptosis rate and concentrations of IL-1β, IL-6, IL-18 and TNF-α in the supernatant were decreased, the expression of ZBP1, caspase-1 and GSDMD was down-regulated ( P<0.05), and no significant change was found in the expression of ZBP1 in group LPS-ATP+ nec-1 ( P>0.05). Conclusions:Activation of ZBP1/RIPK1 signaling pathway is involved in LPS-ATP-induced pyroptosis in macrophages of mice.

Objective:To evaluate the association between N-acylsphingosine amide hydrolase 1 (ASAH1) and pyroptosis during acute lung injury (ALI) in septic mice.Methods:Forty SPF-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-23 g, were divided into 4 groups ( n=10 each) by a random number table method: sham operation group (Sham group), ALI group, HCFU solvent+ ALI group (HA group) and ASAH1 inhibitor HCFU+ ALI group (AA group). The abdominal cavity was only opened in Sham group, and cecal ligation puncture was performed in ALI, HA and AA groups. HCFU solvent 0.2 ml was intraperitoneally injected at 2 h before operation in HA group, and HCFU 10 mg/kg was intraperitoneally injected at 2 h before operation in AA group. The mice were sacrificed at 24 h under deep anesthesia, the eyeballs were removed to collect the blood, the bronchoalveolar lavage fluid (BALF) was collected, and lung tissues and blood samples were collected for microscopic examination of the pathological changes (using HE staining) which were scored and for determination of concentrations of protein in BALF (by BCA method), concentrations of interleukin-1beta (IL-1β), IL-6 and tumor necrosis factor-alpha (TNF-α) in BALF (by enzyme-linked immunosorbent assay), and expression of NOD-like receptor thermoprotein structural domain-related protein 3 (NLRP3) in lung tissues (by Western blot), gasdermin D protein (GSDMD), ASAH1 and cysteine protease-1 (caspase-1) (by Western blot). The wet/dry lung weight (W/D) ratio was calculated. Results:Compared with Sham group, the lung injury score, W/D ratio and concentrations of protein in BALF, IL-1β, IL-6 and TNF-α in serum were significantly increased, and the expression of NLRP3, GSDMD and caspase-1 in lung tissues was up-regulated in ALI, HA and AA groups, and the expression of ASAH1 was significantly up-regulated in ALI and HA groups ( P<0.05). Compared with ALI and HA groups, the lung injury score, W/D ratio, and concentrations of protein in BALF, IL-1β, IL-6 and TNF-α in serum were significantly increased, the expression of NLRP3, GSDMD and caspase-1 in lung tissues was up-regulated, and the expression of ASAH1 was down-regulated in AA group ( P<0.05 or 0.01). Conclusions:ASAH1 is involved in the endogenous protective mechanism underlying ALI in septic mice, which may be related to the inhibition of cell pyroptosis.

Objective:To analyze the mutations of globin genes among patients suspected for thalassemia from the Shanghai area.Methods:A total of 4 644 patients diagnosed at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine between June 2016 and December 2019 were selected as the study subjects. The patients were tested for common mutations associated with thalassemia gene by Gap-PCR and reverse dot blotting (RDB). Patients were suspected to harbor rare mutations based on the inconsistency between hematological phenotypes and results of common mutation detection, and were further analyzed by Gap-PCR and Sanger sequencing.Results:Among the 4 644 patients, 2 194 (47.24%) were found to carry common thalassemia mutations, among which 701 (15.09%) were α-thalassemia, 1 448 (31.18%) were β-thalassemia, and 45 (0.97%) were both α- and β-thalassemia. Forty six samples were found to harbor rare mutations, which included 17 α-globin gene and 29 β-globin gene mutations. CD77(CCC>ACC) ( HBA2: c. 232C>A) of the α-globin gene, NG_000007.3: g. 70567_71015del449, codon 102(-A) ( HBB: c. 308_308delA) and IVS-Ⅱ-636 (A>G) ( HBB: c. 316-215A>G) of the β-globin gene were previously unreported new types of globin gene mutations. Conclusion:Among the 4 644 patients, the detection rate for common thalassemia mutations was 47.24%, whilst 46 samples were detected with rare gene mutations. The type of gene mutation types were diverse in the Shanghai area. The study has provided more accurate results for genetic diagnosis and counseling.