AIM: To investigate and analyze serum nitric oxide synthase 4(NOX4)and matrix metalloproteinase(MMP)-2 expressions in patients with diabetes mellitus(DM)complicated with neovascular glaucoma(NVG)and their clinical significance.METHODS: A prospective study was conducted on 161 patients with DM complicated with NVG admitted to Handan City Eye Hospital(The Third Hospital of Handan)from June 2020 to June 2023. Based on whether complications occurred 1 a after trabeculectomy in the study group patients, they were divided into a group with good prognosis(n=90)and a group with poor prognosis(n=71). During the same period, 161 patients with chronic angle-closure glaucoma without iris neovascularization were selected as the control group. ELISA method was applied to detect the expression levels of serum NOX4 and MMP-2. ROC curve was plotted to evaluate the diagnostic value and postoperative complication prediction value of NOX4 and MMP-2 in patients with DM complicated with NVG. Pearson method was applied to analyze the correlation of NOX4 and MMP-2 with vascular endothelial growth factor(VEGF)and interleukin-6(IL-6). Multivariate Logistic regression was applied to analyze the influencing factors of postoperative complications in the study group.RESULTS:The general information of the study group and control group of patients is comparable. Compared with the control group, the expression levels of serum NOX4, MMP-2, VEGF, and IL-6 in the study group were significantly increased(P<0.001). According to Pearson analysis, serum NOX4 and MMP-2 levels significantly positively correlated with VEGF and IL-6 levels, respectively(P<0.001). According to ROC curve, the AUC of NOX4 combined with MMP-2 in the diagnosis of DM complicated with NVG was better than that of individual diagnosis of NOX4(Z=3.341, P<0.05)and MMP-2(Z=2.788, P<0.05). The duration of DM, the proportion of people with intraocular pressure >21 mmHg, and the expression levels of NOX4, MMP-2, VEGF, and IL-6 in the poor prognosis group were all higher than those in the good prognosis group(P<0.001). The duration of DM, intraocular pressure >21 mmHg, the levels of NOX4, MMP-2, VEGF, and IL-6 were all risk factors for poor prognosis in DM patients with NVG(P<0.001). According to ROC curve, the combined prediction of NOX4 and MMP-2 for postoperative complications in patients with DM complicated by NVG was superior to the AUC predicted by NOX4(Z=3.727, P<0.05)and MMP-2(Z=2.219, P<0.05), respectively.CONCLUSION:NOX4 and MMP-2 are upregulated in the serum of patients with DM complicated by NVG. Combined detection of these two markers holds significant clinical value for the early diagnosis and prognosis of patients with DM complicated by NVG.

BACKGROUND:The femorofibular angle is a commonly used reference angle for mechanical axis correction in high tibial osteotomy on a single plane,but there has been no in-depth study on the application of this angle in high tibial osteotomy on a single plane.OBJECTIVE:To explore the application effect of the femorofibular angle as a reference for mechanical axis correction in high tibial osteotomy on a single plane.METHODS:A retrospective analysis was conducted on 64 cases of knee osteoarthritis patients who underwent high tibial osteotomy on a single plane at Third Affiliated Hospital of Beijing University of Chinese Medicine from January 2023 to January 2024.The patients were grouped according to the intraoperative mechanical axis correction reference method.The observation group consisted of 32 cases,which used the femorofibular angle as a reference for mechanical axis correction during surgery.The control group consisted of 32 cases,which used electrocautery line measurement as a reference for mechanical axis correction during surgery.Both groups received the same perioperative treatment and management.The lower limb mechanical axis ratio,hip-knee-ankle angle,proximal medial tibial angle,and femorofibular angle were recorded before and after surgery to assess changes in the lower limb mechanical state.The operation time and number of fluoroscopies were recorded.The visual analog scale score and Lysholm score for pain and knee function were recorded before surgery and at 1 and 3 months after surgery to evaluate changes in knee pain and function.RESULTS AND CONCLUSION:(1)All patients completed the treatment and follow-up,and no serious adverse events occurred.(2)Before surgery,there was no statistically significant difference in the lower limb mechanical axis ratio,proximal medial tibial angle,hip-knee-ankle angle,and femorofibular angle between the two groups(P＞0.05).At 3 months after surgery,the lower limb mechanical axis ratio,proximal medial tibial angle,hip-knee-ankle angle,and femorofibular angle were significantly improved in both groups compared to before surgery,with statistically significant differences(P＜0.05).There was no significant difference between the two groups(P＞0.05).(3)During surgery,the operation time and number of fluoroscopies in the observation group were less than those in the control group,with statistically significant differences(P＜0.05).(4)At each follow-up point before and after surgery,there was no statistically significant difference in the visual analog scale score and Lysholm score between the two groups(P＞0.05).With the passage of time after surgery,the visual analog scale score and Lysholm score in both groups were significantly improved compared to before surgery(P＜0.05).(5)It is suggested that the application of the femorofibular angle as a reference for mechanical axis correction in high tibial osteotomy on a single plane can accurately locate the range of lower limb mechanical axis correction,is simple and reliable,and reduces the number of fluoroscopies and operation time compared to traditional electrocautery line measurement.

Objective To evaluate the effect of LifePort combined with polymyxin B in preventing donor-derived infections caused by preservation solution contamination. Methods Clinical data of 110 kidney transplant recipients were retrospectively analyzed. According to the decontamination status of preservation solution, the recipients were divided into the decontamination group (n=62) and the non-decontamination group (n=48). The general data of the two groups were compared, and the preventive effect of polymyxin B on possible donor-derived infections (p-DDI) was analyzed, especially infections associated with multidrug-resistant Gram-negative bacteria (MDR GNB). Results There were no statistically significant differences in baseline data (gender, age, preservation solution contamination status, etc.) between the decontamination group and the non-decontamination group (all P > 0.05). The overall contamination rate of preservation solution was 80.0%, and 68 contaminated samples were with single microorganism and 20 with multiple microorganisms. Coagulase-negative staphylococci, Enterococcus and Klebsiella pneumoniae were the most common microorganisms in the positive samples. Fifteen cases of preservation solution were contaminated by MDR GNB, including 10 cases in the non-decontamination group and 5 cases in the decontamination group, with no statistically significant difference between the two groups (P = 0.053). Postoperative infection-related events occurred in 69 recipients, including 39 cases in the non-decontamination group and 30 cases in the decontamination group, with the incidence rate in the non-decontamination group significantly higher than that in the decontamination group (P < 0.001). Only 10 cases of infections were identified as p-DDI, all of which were positive for preservation solution culture, including 8 cases in the non-decontamination group and 2 cases in the decontamination group (P < 0.05). There were 5 cases of p-DDI related to MDR GNB in the non-decontamination group, while no such cases occurred in the decontamination group (P < 0.05). No adverse reactions related to polymyxin B were observed, and no recipient death or renal allograft dysfunction occurred in either group. Conclusions Adding polymyxin B to the preservation fluid during hypothermic machine perfusion with LifePort before renal transplantation may reduce p-DDI and its potential adverse consequences.

Informed consent constitutes a fundamental ethical principle in medical practice. With the in-depth integration of generative artificial intelligence （AI） represented by Chat Generative Pre-trained Transformer （ChatGPT） with medicine， it has brought revolutionary development to traditional informed consent while also introducing new ethical challenges. ChatGPT offers features such as improving the readability of informed consent content， enhancing its comprehensiveness and accuracy， and increasing the convenience of obtaining informed consent. However， as the application of ChatGPT in informed consent is still in the exploratory stage， it is imperative to proactively and fully consider the accompanying ethical issues， such as information security， liability determination， transparency， and fairness. This paper conducted an ethical analysis on the challenges faced by generative AI， represented by ChatGPT， in the application of informed consent and proposed countermeasures， such as upholding free and fully informed consent， strengthening the balance of rights and obligations in informed consent， and establishing a transparent and fair supervision mechanism. The aim was to promote the ethically compliant， orderly， and controllable development of generative AI in the field of medical informed consent.

BACKGROUND:The femorofibular angle is a commonly used reference angle for mechanical axis correction in high tibial osteotomy on a single plane,but there has been no in-depth study on the application of this angle in high tibial osteotomy on a single plane.OBJECTIVE:To explore the application effect of the femorofibular angle as a reference for mechanical axis correction in high tibial osteotomy on a single plane.METHODS:A retrospective analysis was conducted on 64 cases of knee osteoarthritis patients who underwent high tibial osteotomy on a single plane at Third Affiliated Hospital of Beijing University of Chinese Medicine from January 2023 to January 2024.The patients were grouped according to the intraoperative mechanical axis correction reference method.The observation group consisted of 32 cases,which used the femorofibular angle as a reference for mechanical axis correction during surgery.The control group consisted of 32 cases,which used electrocautery line measurement as a reference for mechanical axis correction during surgery.Both groups received the same perioperative treatment and management.The lower limb mechanical axis ratio,hip-knee-ankle angle,proximal medial tibial angle,and femorofibular angle were recorded before and after surgery to assess changes in the lower limb mechanical state.The operation time and number of fluoroscopies were recorded.The visual analog scale score and Lysholm score for pain and knee function were recorded before surgery and at 1 and 3 months after surgery to evaluate changes in knee pain and function.RESULTS AND CONCLUSION:(1)All patients completed the treatment and follow-up,and no serious adverse events occurred.(2)Before surgery,there was no statistically significant difference in the lower limb mechanical axis ratio,proximal medial tibial angle,hip-knee-ankle angle,and femorofibular angle between the two groups(P＞0.05).At 3 months after surgery,the lower limb mechanical axis ratio,proximal medial tibial angle,hip-knee-ankle angle,and femorofibular angle were significantly improved in both groups compared to before surgery,with statistically significant differences(P＜0.05).There was no significant difference between the two groups(P＞0.05).(3)During surgery,the operation time and number of fluoroscopies in the observation group were less than those in the control group,with statistically significant differences(P＜0.05).(4)At each follow-up point before and after surgery,there was no statistically significant difference in the visual analog scale score and Lysholm score between the two groups(P＞0.05).With the passage of time after surgery,the visual analog scale score and Lysholm score in both groups were significantly improved compared to before surgery(P＜0.05).(5)It is suggested that the application of the femorofibular angle as a reference for mechanical axis correction in high tibial osteotomy on a single plane can accurately locate the range of lower limb mechanical axis correction,is simple and reliable,and reduces the number of fluoroscopies and operation time compared to traditional electrocautery line measurement.

Objective To establish an epirubicin (EPI)-resistant murine triple-negative breast cancer (TNBC) (4T1/EPI) cell line and evaluate its biological characteristics and drug resistance. Methods The EPI-resistant cell line 4T1/EPI was developed through intermittent induction with gradually increasing EPI concentrations in vitro. Morphological changes were observed under an inverted microscope. Drug resistance index (MTT assay), cell doubling time (CCK-8 assay), and migration ability (wound healing assay) were evaluated. Western blot was used to detect the expression of drug resistance-related proteins. Transcriptome sequencing and KEGG pathway enrichment analysis were performed to identify the pathways and targets involved in EPI resistance, followed by experimental validation. Results The 4T1 cells eventually grew normally in a medium containing 100 ng/mL EPI, confirming the establishment of the 4T1/EPI resistant cell line. After stable resistance was acquired, morphological alterations were observed. Compared with their parental 4T1 cells, 4T1/EPI cells showed significantly prolonged doubling time (P<0.01) and enhanced migration ability (P<0.05). Expression levels of drug resistance-related proteins MDR1, MRP1 (P<0.01), and ABCG2 (P<0.05) were elevated in 4T1/EPI cells. In vivo models also demonstrated significant EPI resistance in 4T1/EPI tumors in terms of tumor weight and volume. Transcriptome sequencing highlighted the involvement of the PI3K/Akt signaling pathway and ABC transporter pathway. Validation experiments showed the upregulation of Erbb3, Egfr, PI3K, and Akt (P<0.05) and significant downregulation of Fgfr1 (P<0.01) in 4T1/EPI cells. Conclusion The EPI-resistant TNBC cell line 4T1/EPI was successfully established, exhibiting significant resistance in vitro and in vivo. The mechanism may involve the EPI-induced upregulation of Egfr and Erbb3, activating the PI3K/Akt pathway and subsequently enhancing ABC transporter expression.

OBJECTIVE To address the management challenges encountered by inpatient pharmacy of our hospital in undertaking “internet-based medicine delivery” （IMD） services， and to improve service efficiency and medication safety. METHODS SWOT analysis method was applied to systematically examine both internal and external factors， formulate comprehensive management strategies， and restructure the service processes. Process optimization included： establishing a closed-loop process for dispensing and returning drugs based on the drug traceability codes； unifying the drug inventory of the inpatient pharmacy with that of IMD， optimizing the management of storage locations； improving the inventory management function of the hospital information system， and adjusting the inventory counting plan and shift scheduling rules. The indicators of the inpatient pharmacy， including inventory structure， dispensing efficiency， inventory checks accuracy rate， the rationality rate of prescription， and consistency rate of human-machine review， were compared before and after the implementation of the strategies to evaluate the implementation effects. RESULTS After implementation， the structure of the drug inventory was significantly optimized. The proportion of drugs approaching their expiration dates decreased by 35.55%， the proportion of unsold drugs decreased by 64.52%， and the proportion of drugs that had been accumulated for more than 4 weeks decreased by 33.26%. The speed of prescription dispensing increased by 34.92%， and the daily drug requisition time was shortened by 62.03%. The accuracy rate of inventory checks rose from 86.33% to 90.33%， while the rationality rate of prescriptions and the consistency rate of human-machine review rose from 86.00% and 89.33% to 95.00% and 97.00% respectively. Furthermore， the launch of the drug traceability system reduced external dispensing errors from 4 to 1. CONCLUSIONS The comprehensive management strategy based on SWOT analysis can effectively improve the quality of drug management in the inpatient pharmacy and the operational efficiency of IMD， ensuring timely and safe medication for patients.

OBJECTIVE: To investigate the correlation between nucleated red blood cell (NRBC) level on the first day of intensive care unit (ICU) admission and 28-day mortality in adult septic patients, and to evaluate the value of NRBC as an independent predictor of death. METHODS: Single-cell transcriptomic analysis was performed using the GSE167363 dataset from the Gene Expression Omnibus (including 2 healthy controls, 3 surviving septic patients, and 2 non-surviving septic patients). A retrospective clinical analysis was conducted using the America Medical Information Mart for Intensive Care-IV (MIMIC-IV) database, including adult patients (≥ 18 years) with first-time admission who met the Sepsis-3.0 criteria, excluding those without NRBC testing on the first ICU day. The demographic information, vital signs, laboratory test indicators, disease severity score and survival data on the first day of admission were collected. The restricted cubic spline (RCS) curve was used to determine the optimal cut-off value of NRBC for predicting 28-day mortality in patients. Patients were divided into low-risk and high-risk groups based on this cut-off value for intergroup comparison, with Kaplan-Meier survival curve analysis conducted. Independent risk factors for 28-day mortality were analyzed using Logistic regression and Cox regression analysis, followed by the construction of regression models. RESULTS: NRBC were detected in the peripheral blood of septic patients by single-cell transcriptomic. A total of 1 291 sepsis patients were included in the clinical analysis, with 576 deaths within 28 days, corresponding to a 28-day mortality of 44.6%. RCS curve analysis showed a nonlinear relationship between the first-day NRBC level and the 28-day mortality. When NRBC ≥ 1%, the 28-day mortality of patients increased significantly. Compared to the low-risk group (NRBC < 1%), the high-risk group (NRBC ≥ 1%) had significantly higher respiratory rate, heart rate, sequential organ failure assessment (SOFA), and simplified acute physiology score II (SAPSII), and significantly lower hematocrit and platelet count. The high-risk group also had a significantly higher 28-day mortality [49.8% (410/824) vs. 35.5% (166/467), P < 0.05], and shorter median survival time (days: 29.8 vs. 208.6, P < 0.05). Kaplan-Meier survival curve showed that compared with the low-risk group, the survival time of high-risk group was significantly shortened (Log-rank test: χ ² = 25.1, P < 0.001). After adjusting for potential confounding factors including body mass, temperature, heart rate, respiratory rate, mean arterial pressure, serum creatinine, pulse oximetry saturation, hemoglobin, hematocrit, Na⁺, K⁺, platelet count, and SOFA score, multivariate regression analysis confirmed that NRBC ≥ 1% was an independent risk factor for 28-day mortality [Logistic regression: odds ratio (OR) = 1.464, 95% confidence interval (95%CI) was 1.126-1.902, P = 0.004; Cox regression: hazard ratio (HR) = 1.268, 95%CI was 1.050-1.531, P = 0.013]. CONCLUSIONS NRBC ≥ 1% on the first day of ICU admission is an independent risk factor for 28-day mortality in septic patients and can serve as a practical indicator for early prognostic assessment.
Humans ; Sepsis/blood* ; Intensive Care Units ; Risk Factors ; Retrospective Studies ; Prognosis ; Male ; Female ; Hospital Mortality ; Middle Aged ; Aged

The ability of cells to sense and adapt to metabolic changes and environmental variations is essential for their functions. Recent advances in synthetic biology have uncovered increasing mechanisms through which cells detect changes in metabolism and environmental conditions, leading to broader applications. However, a systematic review on the regulation of cellular metabolism and environmental adaption is currently lacking. This article presents a comprehensive overview of this field from three perspectives. First, it introduces key transmembrane and sensor proteins involved in the cellular perception of metabolic and environmental changes. Next, it summarizes the adaptive regulation mechanisms that natural cells employ when confronted with intracellular and extracellular metabolic changes. Finally, the review explores the application scenarios based on cellular adaptive regulation in three aspects: dynamic control, rational metabolic engineering, and adaptive evolution and makes an outlook on the future development directions in this field. This review not only provides a comprehensive perspective on the mechanisms by which cells sense metabolic and environmental variations, but also lays a theoretical foundation for further innovations in the field of synthetic biology. With the continuous advancement of future technologies, a deeper understanding of cellular adaptive regulation mechanisms holds great potential to drive the development and application of novel biomanufacturing platforms.
Adaptation, Physiological ; Synthetic Biology ; Metabolic Engineering/methods* ; Environment ; Bacteria/genetics*

L-valine is an important essential branched-chain amino acid widely used in industries such as feed, pharmaceuticals, and food. In order to further enhance the production performance of L-valine, this study systematically engineered the metabolism of a Corynebacterium glutamicum strain, preserved in the laboratory, which is capable of producing L-valine. First, strain VH-9 was obtained by enhancing the precursor supply, synthesis pathway, and transport system of L-valine. In a 5 L fermenter, the titer, yield, and productivity of L-valine were 76.6 g/L, 0.45 g/g, and 2.39 g/(L·h), respectively. Furthermore, strain VH-18 was obtained by enhancing the uptake of substrate glucose and balancing energy supply to reduce succinate accumulation, with the titer, yield, and productivity of L-valine increased to 82.7 g/L, 0.52 g/g, and 2.58 g/(L·h), respectively. After optimization of fermentation conditions, the titer, yield, and productivity of L-valine in strain VH-18 were further improved to 88.7 g/L, 0.54 g/g, and 2.77 g/(L·h), respectively. This study has achieved the high-efficiency production of L-valine through a systems metabolic engineering strategy.
Corynebacterium glutamicum/genetics* ; Metabolic Engineering/methods* ; Valine/biosynthesis* ; Fermentation ; Glucose/metabolism*