OBJECTIVE To investigate the impact of Netupitant and palonosetron hydrochloride capsules （NEPA） on the pharmacokinetics of Paclitaxel for injection （albumin bound） （i. e. albumin-bound paclitaxel） under different intestinal microenvironment conditions. METHODS Male SD rats were divided into a normal group and a model group （n＝16）. Rats in the model group were intragastrically administered vancomycin solution to establish an intestinal disorder model. The next day after modeling， intestinal microbiota diversity was analyzed， and the mRNA expressions of cytochrome P450 3A1 （CYP3A1） and CYP2C11 in small intestine and liver tissues as well as those protein expressions in liver tissue were measured. Male SD rats were grouped as described above （n＝16）. The normal group was subdivided into the TP chemotherapy group （TP-1 group） and the TP chemotherapy+NEPA group （TP+NEPA-1 group）； the model group was subdivided into the TP chemotherapy group （TP-2 group） and the TP chemotherapy+NEPA group （TP+NEPA-2 group） （n＝8）. Rats in the TP+NEPA-1 and TP+NEPA-2 groups received a single intragastric dose of NEPA suspension （25.8 mg/kg， calculated by netupitant）. One hour later， all four groups received a single tail vein injection of albumin-bound paclitaxel and cisplatin. Blood samples were collected at different time points after the last administration. Using azithromycin as the internal standard， plasma paclitaxel concentrations were determined by liquid chromatography-tandem mass spectrometry. The main pharmacokinetic parameters were calculated using DAS 2.0 software and compared between groups. RESULTS Compared with the normal group， the model group showed significantly decreased Chao1 and Shannon indexes （P＜0.05）， significant alterations in microbiota composition and relative abundance， and significantly downregulated expressions of CYP3A1 mRNA in liver tissue and CYP2C11 mRNA in both small intestine and liver tissues （P＜0.05）. Compared with the TP-1 group， the AUC0－t， AUC0－∞， MRT0－t of paclitaxel in the TP-2 group， the cmax， AUC0－t， AUC0－∞ of paclitaxel in the TP+NEPA-1 group and TP+NEPA-2 group were significantly increased or prolonged； CL of paclitaxel in the TP-2 group， Vd and CL of paclitaxel in the TP+NEPA-1 group and the TP+NEPA-2 group were significantly decreased or shortened （P＜0.05）. Compared with the TP-2 group， cmax of paclitaxel in the TP+NEPA-2 group was significantly increased， and Vd and MRT0－t were significantly decreased or shortened （P＜0.05）. CONCLUSIONS Intestinal microbiota disorder affects the mRNA expressions of CYP3A1 and CYP2C11， leading to decreased clearance and increased systemic exposure of paclitaxel. Concomitant administration of NEPA under normal intestinal microbiota condition increases paclitaxel exposure. However， under conditions of intestinal microbiota disorder， concomitant administration of NEPA has a limited impact on paclitaxel systemic exposure.

The CRISPR/Cas system has emerged as a promising platform for multiplex nucleic acid detection in recent years. Compared with traditional methods such as qPCR and NGS, CRISPR-based assays offer distinct advantages, including simplicity, high sensitivity, and low cost, making them particularly suitable for pathogen screening and early cancer diagnosis. To address the core challenges of CRISPR-based multiplex detection, three types of strategies have been developed: one-pot, spatial separation, and logic gate.These strategies enable simultaneous recognition of multiple targets and effective signal amplification. Applications span viral detection, antimicrobial resistance profiling, and tumor biomarker analysis. Despite persistent challenges such as signal cross interference and system complexity, the technology is evolving rapidly and holds strong potential for clinical translation.

Objective The high post-surgery recurrence rate of endometriosis(EMs)has emerged as a challenge in the long-term manaagement of the condition.This study is aimed at investigating the mechanisms of Neiyiting(NYT)decoction in preventing postoperative recurrence of EMs.Methods An animal model of EMs postoperative recurrence and a model of endometrial stromal cells(hEM15A)cocultured with macrophages(RAW 264.7 cell line)were established for both in vivo and in vitro experiments.An autotransplantation method was used to establish a rat model of EMs.The rats were divided into 4 groups(6 rats per group)and received the corresponding treatments:a Model group receiving distilled water,a Gestrinone group receiving gestrinone at 0.325 mg/kg,a low-dose NYT(NYT-L)group receiving NYT decoction at 5.04 g/(kg-d),and a high-dose NYT(NYT-H)group receiving NYT decoction at 10.08 g/(kg-d).The treatment was administered for 3 weeks via intragastric gavage.In addition,6 SD rats were randomly selected for the control group(Control group),and were given distilled water for 3 weeks via intragastric gavage.The sizes and pathological changes of recurrent lesions in EMs rats were observed.Immunohistochemistry and qRT-PCR were performed to assess the expression of M1 macrophage marker CD86 protein and mRNA in vivo.Additionally,immunohistochemistry and qRT-PCR were used to assess the expression of indicator proteins related to the triggering receptor expressed on myeloid cells 1(TREM1)/Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)signaling pathway and mRNA.The proliferation of hEM15A cells in the coculture experiment was observed.Flow cytometry was performed to determine the polarization of RAW264.7 macrophages,and qRT-PCR was used to determine the expression levels of inducible nitric oxide synthase(iNOS)and interleukin 1β(IL-1β)mRNA.Western blot was performed to determine the expression of signaling pathway-related indicator proteins in vitro.ELISA was performed to determine the levels of inflammatory factors in vitro.Results Compared with the Model group,the volume of recurrent lesions in the NYT-H group was reduced(P＜0.01).Findings from the macrophage M1 polarization assessment showed that the expression levels of CD86 protein and mRNA in the recurrent lesions of the Model group were higher than those in the control group(P＜0.01).The expression levels of CD86 protein and mRNA in the recurrent lesions of the NYT-H group were lower than those of the Model group(P＜0.01).In addition,the RAW 264.7 cell experiment further verified that NYT decoction could reduce the number of CD86-positive macrophages induced by plasmids overexpressing TREM1 and reduce the expression of IL-1β and iNOS mRNA(P＜0.01).The results of the hEM15A cell proliferation assay showed that NYT decoction down-regulated KI-67 protein expression in hEM15A cells induced by macrophage M1 polarization(P＜0.01).The results of TREM1/TLR4/NF-κB signaling pathway showed that the protein and mRNA expression levels of TREM1,TLR4,and NF-κB in the recurrent lesions of the Model group were higher than those of the control group(P＜0.01).Compared with those in the Model group,the protein and mRNA expression levels of TREM1,TLR4,and NF-κB in the recurrent lesions of the NYT-H group were lower(P＜0.01).In addition,the coculture experiment of RAW264.7 and hEM15A cells further confirmed that NYT decoction reduced the expression of TREM1,TLR4,and P-P65 proteins(P＜0.01).Conclusion NYT decoction can inhibit macrophage M1 polarization through the TREM1/TLR4/NF-κB signaling pathway,improve the inflammation level,and inhibit the formation of ectopic endometrial lesions,thereby preventing postoperative recurrence of EMs.

Objective:To assess the accuracy of preoperative enhanced CT in evaluating the degree of lymph node metastasis in gallbladder cancer.Methods:A retrospective analysis was performed on the enhanced CT imaging data of 124 gallbladder cancer patients who underwent surgical treatment at Henan Provincial People's Hospital from Jan 2017 to Dec 2018. Imaging staging was used to classify lymph node metastasis. Pathological and imaging data of 70 patients with confirmed postoperative lymph node pathology were compared to evaluate the accuracy of imaging methods in detecting lymph node involvement.Results:Lymph node metastasis in the 124 surgical patients was categorized into three groups using imaging evaluation methods. The overall accuracy of determining lymph node positivity and negativity was 63%, with a sensitivity of 64% and specificity of 62%. The accuracy of detecting lymph node metastasis in gallbladder cancer was higher when lymph node fusion and internal necrosis were observed. The overall survival rate differed significantly among gallbladder cancer patients at different lymph node imaging stages ( P<0.05). Conclusion:CT imaging evaluation has diagnostic value for lymph node metastasis in gallbladder cancer and has a certain predictive effect on the prognosis of patients.

Objective:To enhance the understanding of clinical doctors on the concurrence of Sj?gren′s syndrome (SS) and myasthenia gravis (MG), provide reference forclinical diagnosis and treatment.Methods:The medical record of SS、MG patient Taian Central hospital in october 2021 was reviewed and a retrospective analysis was conducted. Based on the key words, the main databases domestic and abroad from Jan 1973 to April 2024. The cases reported in the literature were searched out and merged with our case and analyzed by SPSS 27.0. The measurement data of normal distribution were expressed by mean standard deviation ± s, and the measurement data of abnormal distribution were expressed by M( Q1, Q3). Counting data were expressed as examples and percentages (%) . Results:Twenty-one patients with SS complicated with MG were reviewed. Among them, 19(90.5%) patients were female. The average age was (47.6 ± 3.1) years, and the median time from onset to diagnosis was 24 (11, 78) months. There were 11 patients (52.4%) with myasthenia gravis as the initial presentation. The main clinical manifestations of patients with myasthenia muscle fatigue (80.9%, 17/21), gravis include fatigue ptosis (71.4%, 15/21), diplopia, blurred vision (57.1%, 12/21), dysphagia (33.3%, 7/21), joint pain (33.3%, 7/21), odysarthria, and unclear speech (23.8%, 5/21), hair loss (14.3%, 3/21), and swelling of the excocrine glands(9.5%, 2/21). Eighteen patients (85.7%, 18/21) showed positive anti acetylcholine receptor antibodies. Sixteen patients (76.2%) were positive for ANA, and 14 patients (66.8%) were positive for anti SSA or (and) anti SSB antibodies. Four patients (19.0%) also had other autoimmune diseases (Hashimoto′s thyroiditis, rheumatoid arthritis, optic neuritis). In terms of treatment, cholinesterase inhibitors were the most common treatment measure, applied to 18 patients (85.7%), followed by glucocorticoid therapy, and applied to 14 patients (66.8%). In addition, 7 patients (33.3%) received immunosuppressive agents therapy, 5 patients (23.8%) underwent thymectomy, 2 patients (9.5%) received intravenous human immunoglobulin injection, and 1 patient (4.8%) underwent plasma exchange. All patients showed improvement after treatment.Conclusion:SS and MG are both autoimmune diseases, and their coexistence is rare. Clinicans should be aware of this rare association. Early diagnosis is crucial for the treatment and prognosis of patients,and requiresa comprehensive assessment of clinical symptoms, laboratory tests, and auto-antibody test results.

Objective:To explore the characteristics of systemic immune microenvironment during the progression of dextran sulfate sodium(DSS)-induced acute ulcerative colitis(UC)induced in mice by Mass cytometry(CyTOF).Methods:Male C57BL/6 mice were randomly divided into control group and model group.The control group was given normal drinking water for 15 d.The mouse in the model group were given 5%DSS in drinking water,which was changed to normal drinking water after 7 days.In the model group,peripheral blood was collected on days 4,9 and 15,respectively.CyTOF was used to detect the expressions of 33 immune cell markers and changes in cell subsets in peripheral blood of mice,and the characteristics of systemic immune microenvironment in mice with acute UC were analyzed.Results:The cluster analysis of 33 kinds of immune cell markers showed that CD45+cells in peripheral blood of mice with DSS induced acute UC were divided into 23 fine subgroups,among which the proportions of B cell subgroup,T cell subgroup and neutrophil subgroup showed significant changes.A further dimensional reduction cluster analysis of T cell subsets found significant differences in the composition and proportion of the 10 identified T cell subsets.Conclusion:The systemic immune micro-environment map of mice with acute UC induced by DSS has been successfully constructed,and heterogeneity has been found in the systemic immune microenvironment of mice with acute UC.The changes and activation degree of T cell subpopulations are closely re-lated to disease progression and inflammation level.The results of this study provide theoretical basis for assisting the diagnosis,moni-toring the risk,progression,treatment and prognosis of acute UC.

Objective:To identify independent risk factors for early recurrence following curative resection of resectable pancreatic cancer and establish a nomogram prediction model.Methods:Clinical data from 405 patients with resectable pancreatic cancer treated at Renji Hospital, Shanghai Jiao Tong University School of Medicine from February 2010 to December 2020 were retrospectively reviewed. Patients were stratified into a training cohort (265 patients form February 2010 to December 2018) and a validation cohort (140 patients from January 2019 to December 2020) based on surgery dates. Optimal cutoff values for clinical variables were determined using X-tile software. Independent risk factors were identified through univariate and multivariate Cox proportional hazards regression analyses. Kaplan-Meier curves for recurrence-free survival (RFS) were generated across subgroups, and a nomogram was developed to predict early recurrence (within 1 year post-surgery). Time-dependent receiver operating characteristic (tROC) curves was drawn and area under the curve (AUC) metrics were utilized to evaluate predictive accuracy, while model reliability was assessed by calibration curves. Individualized risk scores derived from the nomogram were stratified into high- and low-risk groups using X-tile-derived cutoff values. Survival differences between groups were analyzed via log-rank tests. The clinical application value was judged by decision curve analysis (DCA) compared to TNM staging. Results:In the training cohort, 139 patients (52.45%) experienced early recurrence, with a median RFS of 11.1 months [interquartile range ( IQR): 6.0-26.0]. The validation cohort reported 70 early recurrences (50.00%) and a median RFS of 11.8 months ( IQR: 4.9-21.4). Univariate analysis revealed significant associations between early recurrence and tumor diameter, carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), carbohydrate antigen 125 (CA125), systemic immune-inflammation index (SⅡ), and prognostic nutritional index (PNI). Multivariate analysis identified tumor diameter ≥3.75 cm ( HR=1.718, 95% CI 1.223-2.412, P=0.002), CA19-9≥218 U/ml ( HR=1.567, 95% CI 1.107-2.220, P=0.011), CA125≥20.98 U/ml ( HR=2.501, 95% CI 1.768-3.539, P<0.001), SⅡ≥388.28 ( HR=1.708, 95% CI 1.096-2.662, P=0.018), and PNI<53.18 ( HR=0.596, 95% CI 0.404-0.879, P=0.009) as independent risk factors for early recurrence. The nomogram achieved AUC values of 0.771 and 0.708 in the training and validation cohorts, respectively. Calibration curves demonstrated strong agreement between predicted and observed survival probabilities. Kaplan-Meier analysis revealed significantly lower 1-year RFS rates in high-risk versus low-risk groups for both cohorts (training: HR=3.65, 95% CI 2.45-5.44, P<0.001; validation: HR=2.37, 95% CI 1.39-4.06, P=0.001). DCA indicated superior net benefit of the nomogram over TNM staging across threshold probabilities of 0.2-0.9. Conclusions:The proposed nomogram effectively integrates clinical and serological biomarkers to preoperatively assess early recurrence risk in resectable pancreatic cancer patients, offering enhanced precision for clinical decision-making.

Objective:To explore the characteristics of systemic immune microenvironment during the progression of dextran sulfate sodium(DSS)-induced acute ulcerative colitis(UC)induced in mice by Mass cytometry(CyTOF).Methods:Male C57BL/6 mice were randomly divided into control group and model group.The control group was given normal drinking water for 15 d.The mouse in the model group were given 5%DSS in drinking water,which was changed to normal drinking water after 7 days.In the model group,peripheral blood was collected on days 4,9 and 15,respectively.CyTOF was used to detect the expressions of 33 immune cell markers and changes in cell subsets in peripheral blood of mice,and the characteristics of systemic immune microenvironment in mice with acute UC were analyzed.Results:The cluster analysis of 33 kinds of immune cell markers showed that CD45+cells in peripheral blood of mice with DSS induced acute UC were divided into 23 fine subgroups,among which the proportions of B cell subgroup,T cell subgroup and neutrophil subgroup showed significant changes.A further dimensional reduction cluster analysis of T cell subsets found significant differences in the composition and proportion of the 10 identified T cell subsets.Conclusion:The systemic immune micro-environment map of mice with acute UC induced by DSS has been successfully constructed,and heterogeneity has been found in the systemic immune microenvironment of mice with acute UC.The changes and activation degree of T cell subpopulations are closely re-lated to disease progression and inflammation level.The results of this study provide theoretical basis for assisting the diagnosis,moni-toring the risk,progression,treatment and prognosis of acute UC.

Objective:To identify independent risk factors for early recurrence following curative resection of resectable pancreatic cancer and establish a nomogram prediction model.Methods:Clinical data from 405 patients with resectable pancreatic cancer treated at Renji Hospital, Shanghai Jiao Tong University School of Medicine from February 2010 to December 2020 were retrospectively reviewed. Patients were stratified into a training cohort (265 patients form February 2010 to December 2018) and a validation cohort (140 patients from January 2019 to December 2020) based on surgery dates. Optimal cutoff values for clinical variables were determined using X-tile software. Independent risk factors were identified through univariate and multivariate Cox proportional hazards regression analyses. Kaplan-Meier curves for recurrence-free survival (RFS) were generated across subgroups, and a nomogram was developed to predict early recurrence (within 1 year post-surgery). Time-dependent receiver operating characteristic (tROC) curves was drawn and area under the curve (AUC) metrics were utilized to evaluate predictive accuracy, while model reliability was assessed by calibration curves. Individualized risk scores derived from the nomogram were stratified into high- and low-risk groups using X-tile-derived cutoff values. Survival differences between groups were analyzed via log-rank tests. The clinical application value was judged by decision curve analysis (DCA) compared to TNM staging. Results:In the training cohort, 139 patients (52.45%) experienced early recurrence, with a median RFS of 11.1 months [interquartile range ( IQR): 6.0-26.0]. The validation cohort reported 70 early recurrences (50.00%) and a median RFS of 11.8 months ( IQR: 4.9-21.4). Univariate analysis revealed significant associations between early recurrence and tumor diameter, carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), carbohydrate antigen 125 (CA125), systemic immune-inflammation index (SⅡ), and prognostic nutritional index (PNI). Multivariate analysis identified tumor diameter ≥3.75 cm ( HR=1.718, 95% CI 1.223-2.412, P=0.002), CA19-9≥218 U/ml ( HR=1.567, 95% CI 1.107-2.220, P=0.011), CA125≥20.98 U/ml ( HR=2.501, 95% CI 1.768-3.539, P<0.001), SⅡ≥388.28 ( HR=1.708, 95% CI 1.096-2.662, P=0.018), and PNI<53.18 ( HR=0.596, 95% CI 0.404-0.879, P=0.009) as independent risk factors for early recurrence. The nomogram achieved AUC values of 0.771 and 0.708 in the training and validation cohorts, respectively. Calibration curves demonstrated strong agreement between predicted and observed survival probabilities. Kaplan-Meier analysis revealed significantly lower 1-year RFS rates in high-risk versus low-risk groups for both cohorts (training: HR=3.65, 95% CI 2.45-5.44, P<0.001; validation: HR=2.37, 95% CI 1.39-4.06, P=0.001). DCA indicated superior net benefit of the nomogram over TNM staging across threshold probabilities of 0.2-0.9. Conclusions:The proposed nomogram effectively integrates clinical and serological biomarkers to preoperatively assess early recurrence risk in resectable pancreatic cancer patients, offering enhanced precision for clinical decision-making.

The CRISPR/Cas system has emerged as a promising platform for multiplex nucleic acid detection in recent years. Compared with traditional methods such as qPCR and NGS, CRISPR-based assays offer distinct advantages, including simplicity, high sensitivity, and low cost, making them particularly suitable for pathogen screening and early cancer diagnosis. To address the core challenges of CRISPR-based multiplex detection, three types of strategies have been developed: one-pot, spatial separation, and logic gate.These strategies enable simultaneous recognition of multiple targets and effective signal amplification. Applications span viral detection, antimicrobial resistance profiling, and tumor biomarker analysis. Despite persistent challenges such as signal cross interference and system complexity, the technology is evolving rapidly and holds strong potential for clinical translation.