Objective:To establish a rapid method for the determination of glyphosate and glufosinate residues in plasma samples by solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) .Methods:In March 2024, plasma samples were extracted and protein precipitated with methanol and purified using a weak cation exchange (WCX) SPE cartridge. Then the samples were separated on a Dikma Polyamino HILIC column (150 mm×2.0 mm, 5 μm) with a gradient elution of 1.0 mmol/L ammonium fluoride aqueous solution (pH=11) and acetonitrile as mobile phases. Analysis was performed using an electrospray ionization source (ESI) in multiple reaction monitoring (MRM) mode with internal standard quantification. The method's linearity, detection limits, spiked recovery and precision were then evaluated.Results:Glyphosate and glufosinate exhibited good linearity in the concentration range of 1.0-200.0 μg/L, with correlation coefficients of 0.9986-0.9992. The limits of detection (LODs) of glyphosate and glufosinate were 0.5 and 1.0 μg/L, respectively. At low, medium and high spiked concentrations, the spiked recovery rates and precision for glyphosate and glufosinate in plasma ranged from 92.5% to 113.2% and 3.01% to 11.23%, respectively.Conclusion:The SPE-LC-MS/MS method is simple, rapid, sensitive and accurate, and is suitable for the qualitative and quantitative analysis of glyphosate and glufosinate in plasma from poisoned patients.

Objective:To investigate the effect of olanzapine (OLZ) on the differentially-expressed circRNAs in prefrontal cortex of schizophrenia mouse models and predict the target genes.Methods:SPF grade C57BL/6 mice, 7~8 weeks-old, 20 male mice and 45 female mice were recruited and breeded offspring.Forty-four double-stimulation induced schizophrenia-like mouse models, the offspring mice exposed to dual stress were divided into the schizophrenia group(SZ group, n=23) and the olanzapine intervention group (SZ+ OLZ group, n=21), while the mice raised under normal conditions served as the control group (NC group, n=22). Whole transcriptome sequencing was used to sequence the expression level of RNAs from the prefrontal cortex of the mice. RT-qPCR was applied to verify the differentially-expressed circRNAs, then the target genes of miRNAs which have binding site to verified circRNAs were predicted. Results:RNA-seq results showed that there were 137 differentially-expressed circRNAs compared with NC group, 62 were significantly high-expressed and 75 were low-expressed. circRNA_06886 showed significant low-expressed in SZ group compared with NC group( Z=-3.259, P<0.01), and significant high-expressed in SZ+ OLZ group compared with SZ group( Z=-4.765, P<0.01). Bioinformatics analysis of miRNA target genes showed that the target genes were involved in the pathways related to neural pathways such as dopamine, glutamate and MAPK signaling pathways. Conclusions:There are differentially expressed circRNAs in the prefrontal cortex of schizophrenia mouse models, and circRNA_06886 is low-expressed in the prefrontal cortex of schizophrenia mice, Camk2b-201 and Plcb1-003 are the potential genes of circRNA_06886 involved in the regulation of schizophrenia pathogenesis by dopamine pathway.

Objective:To establish a rapid method for the determination of glyphosate and glufosinate residues in plasma samples by solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) .Methods:In March 2024, plasma samples were extracted and protein precipitated with methanol and purified using a weak cation exchange (WCX) SPE cartridge. Then the samples were separated on a Dikma Polyamino HILIC column (150 mm×2.0 mm, 5 μm) with a gradient elution of 1.0 mmol/L ammonium fluoride aqueous solution (pH=11) and acetonitrile as mobile phases. Analysis was performed using an electrospray ionization source (ESI) in multiple reaction monitoring (MRM) mode with internal standard quantification. The method's linearity, detection limits, spiked recovery and precision were then evaluated.Results:Glyphosate and glufosinate exhibited good linearity in the concentration range of 1.0-200.0 μg/L, with correlation coefficients of 0.9986-0.9992. The limits of detection (LODs) of glyphosate and glufosinate were 0.5 and 1.0 μg/L, respectively. At low, medium and high spiked concentrations, the spiked recovery rates and precision for glyphosate and glufosinate in plasma ranged from 92.5% to 113.2% and 3.01% to 11.23%, respectively.Conclusion:The SPE-LC-MS/MS method is simple, rapid, sensitive and accurate, and is suitable for the qualitative and quantitative analysis of glyphosate and glufosinate in plasma from poisoned patients.

Objective:To investigate the changes and significance of neutrophil-to-lymphocyte ratio, monocyte-to-lymphocyte ratio, and procalcitonin, C-reactive protein, and interleukin-6 levels in neonates with septicemia.Methods:This study used a retrospective analysis mehod. A total of 69 neonates with septicemia who received treatment at the Neonatal Ward, Children's Hospital, The First People's Hospital of Chu Zhou were included in the study group. A total of 43 neonates without septicemia were included in the control group. Neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), and procalcitonin (PCT), C-reactive protein (CRP), and interleukin-6 (IL-6) levels were compared between the two groups. The changes in NLR, MLR, PCT, and CRP levels in neonates with septicemia were observed after treatment, compared with their pre-treatment levels. The receiver operating characteristic curve was used to analyze the value of laboratory indicators NLR, MLR, and PCT, CRP, and IL-6 levels in the diagnosis of septicemia in neonates.Results:The NLR, MLR, and PCT, CRP, and IL-6 levels in the study group were (4.79 ± 1.87), (0.49 ± 0.21), (8.41 ± 3.21) μg/L, (11.97 ± 4.23) mg/L, and (892.13 ± 361.43) ng/L, respectively. These values were significantly higher than those in the control group [(0.89 ± 0.23), (0.13 ± 0.06), (0.51 ± 0.26) μg/L, (2.16 ± 1.26) mg/L, (111.28 ± 51.69) ng/L, t = 10.11, 9.76, 23.06, 36.72, 15.74, all P < 0.05]. After treatment, NLR, MLR, and PCT and CRP levels in the study group were (3.04 ± 1.42), (0.29 ± 0.13), (1.26 ± 0.52) μg/L, and (3.28 ± 1.87) mg/L, respectively. These values were significantly lower than those measured before treatment in the same group [(4.79 ± 2.47), (0.49 ± 0.19), (8.41 ± 3.41) μg/L, (11.97 ± 5.26) mg/L, t = 6.50, 6.33, 33.05, 39.04, all P < 0.05]. The areas under the curve for NLR, MLR, and PCT, CRP, and IL-6 levels in diagnosing septicemia in neonates were 0.785, 0.765, 0.665, 0.763, and 0.753, respectively, with NLR showing the highest area under the curve. Conclusions:NLR and MLR are valuable for diagnosing and evaluating the prognosis of neonatal septicemia. Combined detection of PCT, CRP, and IL-6 levels can enhance the sensitivity and specificity of septicemia diagnosis.

Objective:To investigate the effect of olanzapine (OLZ) on the differentially-expressed circRNAs in prefrontal cortex of schizophrenia mouse models and predict the target genes.Methods:SPF grade C57BL/6 mice, 7~8 weeks-old, 20 male mice and 45 female mice were recruited and breeded offspring.Forty-four double-stimulation induced schizophrenia-like mouse models, the offspring mice exposed to dual stress were divided into the schizophrenia group(SZ group, n=23) and the olanzapine intervention group (SZ+ OLZ group, n=21), while the mice raised under normal conditions served as the control group (NC group, n=22). Whole transcriptome sequencing was used to sequence the expression level of RNAs from the prefrontal cortex of the mice. RT-qPCR was applied to verify the differentially-expressed circRNAs, then the target genes of miRNAs which have binding site to verified circRNAs were predicted. Results:RNA-seq results showed that there were 137 differentially-expressed circRNAs compared with NC group, 62 were significantly high-expressed and 75 were low-expressed. circRNA_06886 showed significant low-expressed in SZ group compared with NC group( Z=-3.259, P<0.01), and significant high-expressed in SZ+ OLZ group compared with SZ group( Z=-4.765, P<0.01). Bioinformatics analysis of miRNA target genes showed that the target genes were involved in the pathways related to neural pathways such as dopamine, glutamate and MAPK signaling pathways. Conclusions:There are differentially expressed circRNAs in the prefrontal cortex of schizophrenia mouse models, and circRNA_06886 is low-expressed in the prefrontal cortex of schizophrenia mice, Camk2b-201 and Plcb1-003 are the potential genes of circRNA_06886 involved in the regulation of schizophrenia pathogenesis by dopamine pathway.

Objective:To investigate the changes and significance of neutrophil-to-lymphocyte ratio, monocyte-to-lymphocyte ratio, and procalcitonin, C-reactive protein, and interleukin-6 levels in neonates with septicemia.Methods:This study used a retrospective analysis mehod. A total of 69 neonates with septicemia who received treatment at the Neonatal Ward, Children's Hospital, The First People's Hospital of Chu Zhou were included in the study group. A total of 43 neonates without septicemia were included in the control group. Neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), and procalcitonin (PCT), C-reactive protein (CRP), and interleukin-6 (IL-6) levels were compared between the two groups. The changes in NLR, MLR, PCT, and CRP levels in neonates with septicemia were observed after treatment, compared with their pre-treatment levels. The receiver operating characteristic curve was used to analyze the value of laboratory indicators NLR, MLR, and PCT, CRP, and IL-6 levels in the diagnosis of septicemia in neonates.Results:The NLR, MLR, and PCT, CRP, and IL-6 levels in the study group were (4.79 ± 1.87), (0.49 ± 0.21), (8.41 ± 3.21) μg/L, (11.97 ± 4.23) mg/L, and (892.13 ± 361.43) ng/L, respectively. These values were significantly higher than those in the control group [(0.89 ± 0.23), (0.13 ± 0.06), (0.51 ± 0.26) μg/L, (2.16 ± 1.26) mg/L, (111.28 ± 51.69) ng/L, t = 10.11, 9.76, 23.06, 36.72, 15.74, all P < 0.05]. After treatment, NLR, MLR, and PCT and CRP levels in the study group were (3.04 ± 1.42), (0.29 ± 0.13), (1.26 ± 0.52) μg/L, and (3.28 ± 1.87) mg/L, respectively. These values were significantly lower than those measured before treatment in the same group [(4.79 ± 2.47), (0.49 ± 0.19), (8.41 ± 3.41) μg/L, (11.97 ± 5.26) mg/L, t = 6.50, 6.33, 33.05, 39.04, all P < 0.05]. The areas under the curve for NLR, MLR, and PCT, CRP, and IL-6 levels in diagnosing septicemia in neonates were 0.785, 0.765, 0.665, 0.763, and 0.753, respectively, with NLR showing the highest area under the curve. Conclusions:NLR and MLR are valuable for diagnosing and evaluating the prognosis of neonatal septicemia. Combined detection of PCT, CRP, and IL-6 levels can enhance the sensitivity and specificity of septicemia diagnosis.

Objective:To explore the role of arginine metabolism in the inflammatory response to influenza A virus (FluA) infection.Methods:Eighteen-month-old mice were infected with FluA via nasal drip, with samples collected on the 6th day post-infection. The concentration of cytokines was determined by the Luminex multifactor assay, while the metabolites in bronchoalveolar lavage fluid (BALF) were analyzed using targeted metabolomic method. Correlation analysis was employed to examine the correlation between cytokines and metabolites. Macrophages were infected with FluA at a multiplicity of infection (MOI) of 1 and cultured with different concentrations of arginine for 24 h. The mRNA and protein expression levels of interleukin (IL)-1β, IL-6, tumor necrosis factor α (TNF-α) and IL-10 were detected by real time fluorescence quantitative RT-PCR (qRT-PCR) and Cytometric Bead Array (CBA).Results:In comparison to the control group, the levels of surfactant protein D (SP-D), TNF-α, monocyte chemotactic protein-1 (MCP-1), IL-1α, IL-6, IL-10, recombinant S100 calcium binding protein (S100) A9, interferon inducible protein 10 (IP-10), macrophage inflammatory protein-1α (MIP-1α), matrix metalloprotein-9 (MMP-9), and Complement Factor D in BALF of FluA infection exhibited a significant elevation. The concentrations of arginine, aspartate, citrulline, glutamic acid, ornithine, proline, creatine, and sarcosine in arginine metabolism were up-regulated, which was correlated with most of elevated cytokine levels. The supplementation of arginine after FluA infection significantly decreased the levels of IL-1β, IL-6, and TNF-α, but increased the level of IL-10 in macrophages.Conclusions:Arginine reduces the inflammatory response induced by FluA infection in macrophages, suggesting that it may be a potential intervention target for severe pulmonary inflammation following FluA infection.

Objective:To evaluate the effect of hyperthermia on radiation pneumonitis (RP) in elderly patients with esophageal cancer undergoing intensity-modulated radiotherapy (IMRT).Methods:Clinical data of 177 elderly esophageal cancer patients (aged ≥60 years) receiving IMRT in the First Affiliated Hospital of Soochow University and Yixing Cancer Hospital from August 1, 2017 to February 6, 2023 were retrospectively analyzed. Patients were divided into the hyperthermia and non-hyperthermia groups based on whether they received hyperthermia treatment. Patients in two groups received IMRT with 6 MV X-rays. Patients in the hyperthermia group underwent high-frequency hyperthermia within 1 h before radiation using the external thermotherapy device HG-2000Ⅲ (heating temperature: 41-43 ℃ for 40 min, twice a week). After adjusting for confounding factors between two groups using propensity score matching (PSM), the short-term effective rates between two groups were compared using Chi-square test. Univariate analysis and logistic multivariate analysis were employed to compare the incidence of RP between two groups. Results:After applying PSM, 42 pairs were successfully matched, and the baseline data and radiotherapy parameters showed no statistically significant differences between two groups (all P>0.05). The objective response rate (ORR) in the hyperthermia group was significantly higher than that in the non-hyperthermia group (83.3% vs. 64.3%, P=0.047). Univariate analysis revealed that the incidence of RP and symptomatic RP (≥ grade 2) in the hyperthermia group was significantly lower than that in the non-hyperthermia group (61.9% vs. 85.7%, P=0.013; 21.4% vs. 47.6%, P=0.012). Logistic multivariate analysis indicated that hyperthermia was an independent protective factor for symptomatic RP ( P=0.011). Conclusions:The incidence and severity of RP in elderly esophageal cancer patients receiving IMRT can be reduced by hyperthermia. Hyperthermia, as a clinically beneficial green treatment, improves efficacy and reduces toxicity for patients with esophageal cancer.

Objective:To compare the prognosis of patients with hepatolithiasis-associated intrahepatic cholangiocarcinoma (ICC) and hepatitis B virus (HBV)-associated ICC after radical resection.Methods:The clinicopathological and survival data of 69 patients with ICC undergoing radical resection in the Department of Hepatobiliary and Pancreatic Surgery, Lihuili Hospital Affiliated to Ningbo University from January 2012 to February 2024 were retrospectively analyzed, including 40 males and 29 females, aged (62.9±10.2) years. Patients were divided into the stone group ( n=40, with hepatolithiasis) and HBV group ( n=29, with HBV). Kaplan-Meier method was used for survival analysis, and log-rank test was used for survival rate comparison. Cox proportional hazard regression model was used for multivariate analysis to analyze the effect of hepatolithiasis and HBV on the prognosis. Results:The median overall survival of ICC patients in the stone group was 16 months, and that in the HBV group was 27 months. The 1-, 3-, and 5-year cumulative survivals of the stone group were 56.6%, 23.2%, and 10.3%, respectively, which were lower than those of the HBV group (72.0%, 50.7%, and 43.4%, respectively, χ2=5.95, P=0.015). The median recurrence-free survivals (RFS) of the stone group and the HBV group were 12 months and 23 months, respectively. The 1-year and 3-year RFS of the stone group were 49.2% and 18.1%, which were lower than those of the HBV group (65.0% and 39.8%, respectively, χ2=3.94, P=0.047). Univariate analysis showed that hepatolithiasis was assciated with prognosis ( χ2=5.95, P=0.015). Multivariate Cox regression analysis showed that hepatolithiasis and hepatitis B virus infection had no effect on the prognosis of ICC patients after surgery (all P>0.05). Conclusion:Compared to HBV infection, ICC patients with hepatolithiasis have a worse prognosis. Hepatolithiasis and HBV infection have no effect on the prognosis of ICC after radical resection.

Objective:To investigate the clinicopathologic features of sarcomatoid carcinoma of the pancreas.Methods:The clinicopathological data of 7 cases with sarcomatoid carcinoma of the pancreas admitted in the Affiliated Lihuili Hospital of Ningbo University from September 2013 to August 2021 were retrospectively analyzed, including clinical manifestations, laboratory examination, imaging examination, pathological examination of tissue specimens, surgical methods and adjuvant treatments. Expressions of mesenchymal markers and epithelial markers in tumor tissues were determined by immunohistochemical staining.Results:Among the 7 cases of sarcomatoid carcinoma of the pancreas, there were 4 male and 3 female. The patient age ranged from 51 to 88 years old, and the mean age was 69 years old. All the patients underwent CT examimation before surgery. 3 tumors were located in the head, 3 in the body and 1 in the tail of the pancreas. CT examination also showed that 4 tumors were cystic solid and 3 were cystic. Six patients underwent radical surgery and one underwent partial resection for biopsy. Microscopically, the tumor was predominantly composed of sarcomatoid spindle-shaped cells. Immunohistochemical staining showed that the tumor expressed both mesenchymal markers vimentin and epithelial marker CK7, CK19, CK(pan) and CAM5.2. The overall prognosis of the patients was poor, 4 cases died within 1 year after surgery, and the other 3 cases survived without recurrence.Conclusions:The clinical manifestations of sarcomatoid carcinoma of the pancreas were not typical, but the pathological and immunohistochemical features are obvious and the prognosis is poor.