OBJECTIVE To establish a method for simultaneous determination of ceftazidime， avibactam and aztreonam concentrations in human plasma.METHODS Stable-isotope-labeled compounds ceftazidime-D 5 ， avibactam- 13 C 5 and aztreonam- 2 H 6 were used as internal standards. Plasma samples were pretreated by protein precipitation with methanol， followed by nitrogen blowing. A ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） technique was applied to develop a quantitative analytical method for simultaneous measurement of ceftazidime， avibactam and aztreonam in human plasma. An ACQUITY UPLC BEH C 18 column was used as the chromatographic column. Gradient elution was performed with 0.01% formic acid-acetonitrile solution as the mobile phase at a flow rate of 0.3 mL/min. The column temperature was set at 40 ℃， and the injection volume was 5 μL. An electrospray ionization source was operated in both positive and negative ion modes for multiple reaction monitoring. Method validation was carried out according to the Guidelines for Bioanalytical Method Validation contained in volume Ⅳ of the 2020 edition of Chinese Pharmacopoeia .RESULTS The linear ranges of ceftazidime， avibactam and aztreonam were 0.04-200， 0.05-50，0.01-50 μg/mL， respectively， with all correlation coefficients no less than 0.996. The relative error （RE） for accuracy was no more than 12.25% （ n ＝5）， and the RSD for precision was no more than 13.49% （ n ＝5）. The RSD of internal-standard-normalized matrix factors was less than 15.00% （ n ＝6）. The RE in stability tests was lower than 15.00% （ n ＝3）.CONCLUSIONS This study successfully established an UPLC-MS/MS method for simultaneous determination of ceftazidime， avibactam and aztreonam concentrations in human plasma. The method possesses favorable accuracy， precision and stability， and can provide methodological support for therapeutic drug monitoring and individualized medication in patients treated with these three combined drugs.

Objective:To evaluate the clinical efficacy of Buyang Huanwu Decoction combined with medium-frequency pulsed electrical stimulation in the treatment of diabetic peripheral neuropathic pain (DPNP).Methods:A randomized controlled trial study was conducted. A total of 86 patients with DPNP in the endocrinology department of Xi'an Hospital of Traditional Chinese Medicine from September 2020 to September 2023 were selected as the observation objects and divided into 2 groups through the random number table method, with 43 cases in each group. The control group was combined with medium-frequency pulsed electrical stimulation on the basis of conventional Western medical treatment, while the observation group was supplemented with Buyang Huanwu Decoction on the basis of the control group. Both groups were treated for 12 weeks. TCM syndrome scores were conducted before and after the treatment respectively. The Toronto Clinical Scoring System (TCSS) was used to grade the severity of neuropathy. The motor nerve conduction velocity (MCV) and sensory nerve conduction velocity (SCV) of the median nerve and common peroneal nerve were detected by electromyography evoked potential tester. Adverse reactions during the treatment were recorded to evaluate the clinical efficacy.Results:The total effective rate was 93.02% (40/43) in the observation group and 74.42% (32/43) in the control group, with statistical significance between the two groups ( χ2=5.46, P=0.019). After treatment, the scores of shortness of breath and fatigue, end-limb pain, listlessness, dull tongue and the total score in the observation group were lower than those in the control group ( t values were 3.06, 3.42, 3.09, 3.57 and 3.98 respectively, P<0.05); the scores of neurological symptoms, neural reflexes and sensory function examinations were lower than those in the control group ( t values were 4.54, 6.28 and 5.16 respectively, P<0.01). After treatment in the observation group, MCV of the median nerve [(47.96±5.98) m/s vs. (42.02±5.74) m/s, t=4.70] and SCV [(54.90±6.29) m/s vs. (48.04±6.16) m/s, t=5.11] were higher than those in the control group. The MCV of the common perbular nerve [(41.76±6.27) m/s vs. (37.04±6.18) m/s, t=3.52] and SCV [(50.35±6.41) m/s vs. (46.14±6.13) m/s, t=3.12] were higher than those in the control group ( P<0.01 or P<0.05). During the treatment period, the incidence of adverse reactions was 11.63% (5/43) in the observation group and 9.30% (4/43) in the control group, without statistical significance between the two groups ( χ2=0.12, P=0.725). Conclusion:Buyang Huanwu Decoction combined with medium-frequency pulsed electrical stimulation can effectively improve the degree of neuropathy in patients with DPNP, increase nerve conduction velocity, alleviate TCM syndromes, enhance clinical efficacy and has good safety.

Objective:To explore the association between sublingual microcirculatory perfusion vessel proportion (PPV), high mobility group protein B1 (HMGB1), and sepsis-induced coagulopathy (SIC), and to identify early predictive markers for clinical intervention.Methods:A total of 66 septic patients admitted to the Respiratory Intensive Care Unit (RICU) between November 2021 and May 2024 were enrolled. Based on SIC diagnosis within 24 hours of admission, patients were categorized into SIC ( n=36) and non-SIC ( n=30) groups. Clinical parameters, including prothrombin time (PT), activated partial thromboplastin time (APTT), sublingual PPV, and serum HMGB1 levels (measured at 0 h and 6 h), were analyzed. Logistic regression and receiver operating characteristic (ROC) curve analyses were performed to identify independent predictors. Results:Compared with the non-SIC group, the SIC group demonstrated significantly prolonged PT ( P<0.05) and APTT ( P<0.05), elevated 0 h lactate levels, and increased 6 h HMGB1. Spearman analysis revealed a positive correlation between 6 h PPV and platelet count ( P<0.05), while an inverse correlation was observed with 6 h PT ( P<0.05). Multivariate regression identified APTT, 6 h PPV, and 6 h HMGB1 as independent predictors of SIC. ROC analysis yielded AUC values of 0.732 for APTT, 0.802 for 6 h PPV, and 0.765 for 6 h HMGB1. The combination of 6 h PPV and HMGB1 further improved predictive accuracy (AUC=0.873). Conclusions:Prolonged APTT, decreased 6 h PPV, and elevated 6h HMGB1 are key indicators of SIC. Integrated assessment of these markers enhances early risk stratification in septic patients, facilitating timely clinical intervention.

Objective To investigate the correlation between the levels of activated protein C(APC),thromboxane B2(TXB2),and soluble B7-H3(sB7-H3)in the peripheral blood of elderly patients with pneumonia and the severity as well as prognosis of the disease.Methods One hundred elderly pneumonia patients admitted to our hospital from March 2022 to June 2024 were enrolled as the study group,and 100 healthy volunteers during the same period were selected as the control group.The levels of APC,TXB2,and sB7-H3 in peripheral blood were com-pared between the two groups.Study group patients were further categorized into low-risk,medium-risk,and high-risk subgroups based on the CURB-65 score(Confusion,Uremia,Respiratory rate,Blood pressure,Age≥65 years).The levels of APC,TXB2,and sB7-H3 in peripheral blood were compared among patients with varying disease severities.Pearson correlation analysis was performed to evaluate the correlation between the levels of APC,TXB2,and sB7-H3 in peripheral blood and disease severity.The study group was followed up for 30 days and subsequently divided into good prognosis and poor prognosis subgroups according to their clinical outcomes.Clinical data and peripheral blood levels of APC,TXB2,and sB7-H3 were compared between patients with different prognoses.Partial correlation analysis was conducted to assess the relationship between peripheral blood levels of APC,TXB2,and sB7-H3 and prognosis.Finally,the predictive value of these biomarkers was evaluated using the Receiver Oper-ating Characteristic(ROC)curve.Results The level of APC in the peripheral blood of the study group was signifi-cantly lower than that of the control group,whereas the levels of TXB2 and sB7-H3 were significantly higher(P<0.05).The severity of the disease in the study group was assessed using the CURB-65 score,which categorized patients into 33 mild cases,39 moderate cases,and 28 severe cases.Severe patients exhibited a lower level of APC in peripheral blood compared to moderate and mild patients.Additionally,the levels of TXB2 and sB7-H3 in moderate patients were higher than those in mild patients,while severe patients demonstrated even higher levels of TXB2 and sB7-H3 compared to both moderate and mild patients(P<0.05).Pearson correlation analysis revealed that peripheral blood APC was negatively correlated with the CURB-65 score,whereas TXB2 and sB7-H3 were positively correlated with the CURB-65 score(P<0.05).During a 30-day follow-up period,the research team identified 79 patients with good prognoses and 21 patients with poor prognoses.Significant differences were observed in diabetes prevalence,disease severity,and APACHEⅡ scores between the poor prognosis subgroup and the good prognosis subgroup(P<0.05).The levels of APC in peripheral blood were significantly lower in the poor prognosis subgroup compared to the good prognosis subgroup,whereas the levels of TXB2 and sB7-H3 were significantly higher(P<0.05).Partial correlation analysis revealed that peripheral blood APC,TXB2,and sB7-H3 were significantly associ-ated with prognosis(P<0.05).The AUC values for predicting the prognosis of elderly pneumonia patients using peripheral blood APC,TXB2,and sB7-H3 were 0.752,0.738,and 0.761,respectively,with sensitivities of 66.67%,76.19%,and 66.67%,and specificities of 78.48%,67.09%,and 78.48%.When combining these three indicators for prognostic prediction,the AUC increased to 0.918,with a sensitivity of 85.71%and a specificity of 87.34%,demonstrating a significant improvement in predictive accuracy compared to each indicator used alone(Z=2.207,2.666,2.109,P=0.027,0.008,0.035).Conclusion The levels of APC,TXB2,and sB7-H3 in the peripheral blood of elderly patients with pneumonia are significantly associated with the severity and prognosis of the disease.Combined detection of these biomarkers can serve as a reliable predictor of clinical outcomes.

Objective:To investigate the improvement in nutritional indicators, clinical complications, and respiratory infections among children with cerebral palsy (CP) and malnutrition who meet the indications for tube feeding under oral feeding and tube feeding interventions.Methods:In this retrospective cohort study, 82 children with CP and malnutrition that met the indications for tube feeding from the Children′s Rehabilitation Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2022 to April 2024 were included.These patients were divided into an oral feeding group and a tube feeding group.The t-test, Mann-Whitney U test, or Kruskal-Wallis test was used to analyze nutritional indices such as weight-for-age Z-score (WAZ), height/length-for-age Z-score (HAZ), and weight-for-height/length Z-score (WHZ) before and after intervention in both groups.The improvement in clinical complications and respiratory infections was also analyzed. Results:After 3-month nutritional intervention, there were 60 cases in the oral feeding group and 22 cases in the tube feeding group.The tube feeding group showed better improvement in WHZ [-0.23(-1.79, 0.88) vs.-1.62(-2.02, -0.91) ] than the oral feeding group ( P<0.05).After 6-month nutritional intervention, there were 50 cases in the oral feeding group and 16 cases in the tube feeding group.The tube feeding group showed better improvement in WAZ [-0.80(-1.92, -0.42) vs. -2.26(-2.88, -1.84)], HAZ (-1.31±1.23 vs. -2.32±1.19), and WHZ (-0.74±1.39 vs. -1.58±1.03) than the oral feeding group (all P<0.05).After 12-month nutritional intervention, there were 30 cases in the oral feeding group and 10 cases in the tube feeding group.The tube feeding group showed better improvement in WAZ [-1.06(-1.77, -0.88) vs.-2.25(-3.06, -1.47) ] and HAZ (-1.22±1.63 vs. -2.54±1.50) than the oral feeding group (all P<0.05).Clinical complications improved better in the tube feeding group than those in the oral feeding group.In the tube feeding group, significant differences in choking ( χ2=16.29, P=0.001) and vomiting ( χ2=6.81, P=0.013) were observed before and after nutritional intervention.There was a statistically significant difference in frequency of lower respiratory infections 3 and 6 months after nutritional intervention between the 2 groups (all P<0.05). Conclusions:Compared with oral feeding, tube feeding can effectively improve the clinical nutritional indicators and reduce the clinical complications and the risk of respiratory infections in CP children that meet the indications for tube feeding.

Objective:Middle ear cholesteatoma is a common otolaryngological disease, and traditional diagnostic methods have certain limitations. This study aims to construct a computer-aided diagnosis model for middle ear cholesteatoma based on integrated convolutional neural networks (CNNs) to improve diagnostic accuracy and efficiency.Methods:Firstly, Data were collected from patients who visited the Department of Otorhinolaryngology Head and Neck Surgery at the First People′s Hospital of Yinchuan between January 2020 and December 2021. 8 000 temporal bone CT images were collected, including 5 000 images diagnosed pathologically as middle ear cholesteatoma and 3 000 normal images. A five-fold cross-validation method was used to divide the dataset into training and testing sets. Next, a transfer learning approach was used to initialize model parameters, and the AlexNet, GoogleNet, and ResNet networks were pre-trained to extract deep features from the images. Then, the Softmax classification algorithm was applied to classify the features, resulting in three independent classifiers. These classifiers were combined using an ensemble learning method with a weighted voting approach to obtain the final diagnostic results. Finally, the model was evaluated by comparing the ensemble classifier with individual classifiers to assess its accuracy, precision, sensitivity, specificity, and diagnostic time, and a comparison with low-mid-and high-experience physician groups was conducted to comprehensively evaluate the model′s diagnostic performance.Results:The experimental results showed that the model achieved an accuracy of 88.8%(178/200), precision of 92.9%,(112/120) sensitivity of 89.8%(108/120), and specificity of 88.1%(70/80). The average diagnostic time for individual patient temporal bone CT images was reduced to 2-3 seconds. Compared to the diagnostic results from low-mid-and high-experience physician groups, the model demonstrated significant advantages and effectively assisted clinicians in making rapid and accurate middle ear cholesteatoma diagnoses.Conclusion:The proposed middle ear cholesteatoma diagnostic model based on integrated convolutional neural networks exhibits high recognition accuracy and rapid diagnostic speed, significantly improving clinical diagnostic efficiency, especially in early screening and auxiliary diagnosis, making it of considerable value in clinical practice.

OBJECTIVES: To investigate the regulatory role of the NLRP3 signaling pathway in hepatocyte pyroptosis in nonalcoholic steatohepatitis (NASH) under hypoxia. METHODS: Twenty-four male C57BL/6 mice were randomized equally into hypoxic control (A), hypoxic NASH model (B), hypoxic NASH+NLRP3 inhibitor (C), and hypoxic NASH+caspase-1 inhibitor (D) groups. In groups B-D, the mice were fed a methionine choline-deficient (MCD) diet under hypoxic conditions (to simulate a 5000 m altitude) for 6 weeks; the mice in groups C and D received intraperitoneal injections of the respective inhibitors every other day. RESULTS: Compared with those in group A, the mice in group B showed significantly elevated serum levels of FBG, TC, TG, ALT and AST, increased liver lipid content, inflammatory cell infiltration and collagen fiber deposition, and enhanced hepatic expressions of NLRP3, caspase-1, IL-1β and GSDMD proteins, with obvious swelling, cristae breakage, vacuolization, and outer membrane disruption of the mitochondria, ribosome loss in the cytoplasm, destruction of the nuclear membrane, and pathological changes of the rough endoplasmic reticulum. Treatment with NLRP3 inhibitor and caspase-1 inhibitor both significantly lowered serum levels of TC, TG, ALT and AST (but without significantly affecting FBG) in the mouse models, and reduced liver lipid content, inflammatory cell infiltration, collagen deposition, and expression levels of NLRP3, caspase-1, GSDMD and IL-1β. The treatments also significantly improved pathological changes in the mitochondria, ribosomes and endoplasmic reticulum in liver tissues of the mice. CONCLUSIONS NLRP3 signaling pathway plays a key role in promoting hepatocyte pyroptosis in NASH mice under hypoxic condition, and inhibiting this pathway can effectively reduce liver inflammation, suggesting its potential as a therapeutic target for NASH treatment.
Animals ; Non-alcoholic Fatty Liver Disease/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Pyroptosis ; Mice, Inbred C57BL ; Male ; Hepatocytes/pathology* ; Signal Transduction ; Mice ; Hypoxia/metabolism* ; Caspase 1/metabolism* ; Interleukin-1beta/metabolism* ; Liver/metabolism*

Objective:To develop an artificial intelligence（AI）-based sequential ultrasound-magnetic resonance imaging（US-MRI）diagnostic strategy to optimize the imaging workflow for ovarian masses.Methods:A total of 1 120 patients with pathologically confirmed ovarian masses who underwent both preoperative pelvic ultrasound and MRI between January 2021 and December 2023 at Women's Hospital，Zhejiang University School of Medicine were retrospectively included. Patients were randomly divided into the training（ n=672）and internal test set（ n=448）at a ratio of 6∶4. An external test set（ n=128）was established at the Forth Affiliated Hospital of School of Medicine. Deep learning was used for automated segmentation of MRI lesions，followed by radiomic feature extraction and machine learning classification to construct both a US-MRI multimodal model and sequential US-MRI strategy. Diagnostic performance and potential healthcare cost-saving effects were evaluated across strategies. Results:In the internal test set（ n=448），the AI-based sequential US-MRI strategy achieved a F1 score of 0.863 and a diagnostic accuracy of 82.14%，with no significant difference compared to the US-MRI multi-modal model（ P>0.05）. The sequential strategy identified 82 cases（18.30%，82/448）of patients as low-risk true negatives during initial ultrasound screening，suggesting a potential to reduce the need for MRI examinations in future clinical practice. In the external test set（ n=128），the strategy achieved an F1 score of 0.800 and a confirmed diagnosis rate of 85.94%，with a theoretical reduction of 26.56%（34 cases）in MRI utilization while maintaining a diagnostic accuracy rate higher than that of the multi-modal model（82.18%）. Conclusions:The AI-based US-MRI sequential diagnostic strategy demonstrates favorable diagnostic accuracy while offering the potential to optimize MRI utilization. This approach may enhance the efficiency of imaging resource allocation and reduce healthcare burden in the management of ovarian masses.

BACKGROUND: The role of inflammation in the development of gestational diabetes mellitus (GDM) has recently become a focus of research. The systemic immune-inflammation index (SII) and systemic inflammation response index (SIRI), novel indices, reflect the body's chronic immune-inflammatory state. This study aimed to investigate the associations between the SII or SIRI and GDM. METHODS: A prospective birth cohort study was conducted at Beijing Obstetrics and Gynecology Hospital from February 2018 to December 2020, recruiting participants in their first trimester of pregnancy. Baseline SII and SIRI values were derived from routine clinical blood results, calculated as follows: SII = neutrophil (Neut) count × platelet (PLT) count/lymphocyte (Lymph) count, SIRI = Neut count × monocyte (Mono) count/Lymph count, with participants being grouped by quartiles of their SII or SIRI values. Participants were followed up for GDM with a 75-g, 2-h oral glucose tolerance test (OGTT) at 24-28 weeks of gestation using the glucose thresholds of the International Association of Diabetes and Pregnancy Study Groups (IADPSG). Logistic regression was used to analyze the odds ratios (ORs) (95% confidence intervals [CIs]) for the the associations between SII, SIRI, and the risk of GDM. RESULTS: Among the 28,124 women included in the study, the average age was 31.8 ± 3.8 years, and 15.76% (4432/28,124) developed GDM. Higher SII and SIRI quartiles were correlated with increased GDM rates, with rates ranging from 12.26% (862/7031) in the lowest quartile to 20.10% (1413/7031) in the highest quartile for the SII ( Ptrend <0.001) and 11.92-19.31% for the SIRI ( Ptrend <0.001). The ORs (95% CIs) of the second, third, and fourth SII quartiles were 1.09 (0.98-1.21), 1.21 (1.09-1.34), and 1.39 (1.26-1.54), respectively. The SIRI findings paralleled the SII outcomes. For the second through fourth quartiles, the ORs (95% CIs) were 1.24 (1.12-1.38), 1.41 (1.27-1.57), and 1.64 (1.48-1.82), respectively. These associations were maintained in subgroup and sensitivity analyses. CONCLUSION The SII and SIRI are potential independent risk factors contributing to the onset of GDM.
Humans ; Female ; Pregnancy ; Diabetes, Gestational/immunology* ; Prospective Studies ; Adult ; Inflammation/immunology* ; Glucose Tolerance Test ; Birth Cohort

Imprinted genes play a key role in regulating mammalian placental and embryonic development. Here, we generated glutaminyl-peptide cyclotransferase-knockout (Qpct^-/-) mice utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) platform and identified Qpct as a novel anti-angiogenic factor in regulating mouse placentation. Compared with Qpct^+/+ mice, placentae and embryos (Qpct^-/+ and Qpct^-/-) showed significant overgrowth at embryonic Day 12.5 (E12.5), E15.5, and E18.5. Using single-cell transcriptome analysis of 32 309 cells from Qpct^+/+ and Qpct^-/- mouse placentae, we identified 13 cell clusters via single-nucleus RNA sequencing (snRNA-seq) (8880 Qpct^+/+ and 13 577 Qpct^-/- cells) and 20 cell clusters via single-cell RNA sequencing (scRNA-seq) (6567 Qpct^+/+ and 3285 Qpct^-/- cells). Furthermore, we observed a global up-regulation of pro-angiogenic genes in the Qpct^-/- background. Immunohistochemistry assays revealed a notable increase in the number of blood vessels in the decidual and labyrinthine layers of E15.5 Qpct^-/+ and Qpct^-/- mice. Moreover, the elevation of multiple pairs of ligand-receptor interactions was observed in decidual cells, endothelial cells, and macrophages, promoting angiogenesis and inflammatory response. Our findings indicate that loss of maternal Qpct leads to altered phenotypic characteristics of placentae and embryos and promotes angiogenesis in murine placentae.
Animals ; Female ; Pregnancy ; Mice ; Placentation/genetics* ; Single-Cell Analysis ; Gene Expression Profiling ; Mice, Knockout ; Transcriptome ; Placenta/blood supply* ; Neovascularization, Pathologic/genetics* ; Genomic Imprinting ; Single-Cell Gene Expression Analysis ; Angiogenesis