[Objective] To explore the accuracy and feasibility of non-invasive prenatal diagnosis of fetal RhE genotype using cell-free fetal DNA (cff-DNA) from maternal peripheral blood. [Methods] A total of 134 pregnant women with single fetuses and RhE-negative blood group were selected from our hospital from November 2023 to August 2024. Free DNA extraction kit was used to extract free DNA from peripheral blood of pregnant women, and the RhE blood group genotype of free DNA was detected by real-time fluorescent quantitative PCR (RT-qPCR). If the qPCR amplification signal of the sample was negative, the methylated RASSF1A gene was amplified, and the positive amplification result was used as a sign of successful extraction of cff-DNA. Serological microcolumn gel method was used to detect the phenotype of RhE blood group in neonatal peripheral blood. [Results] Among the 134 maternal peripheral blood samples, the cff-DNA detection of RhE blood group phenotypes was consistent with the RhE blood group genotyping of neonatal peripheral blood in 133 cases, including 90 cases of Rhee genotype and 43 cases of RhE genotype, with diagnostic concordance rate of 99.3%, sensitivity of 97.7%, specificity of 100%, youden index of 0.977, area under ROC curve of 0.995, the Kappa value of 0.983, positive predictive value of 100%, and negative predictive value of 98.9%. The sample of 1 case failed to be detected. After the amplification of methylated RASSFIA gene, it was confirmed that the reason for the failure was that no cff-DNA was extracted from the sample. The diagnostic concordance rates of the first, second and third trimesters were 93.8% (15/16), 100% (51/51) and 100% (67/67), respectively. Fisher's exact test method was used to calculate the P value, which was P>0.05, indicating that there was no statistical significance in the difference of diagnostic concordance rate among the three pregnancy periods, and there was no difference in the detection concordance rate of this method in different pregnancy periods. [Conclusion] The use of cff-DNA in maternal peripheral blood for the detection of fetal RhE blood group genotype is an accurate and highly feasible non-invasive prenatal diagnostic method, which is helpful for the clinical diagnosis of fetal and neonatal hemolytic disease caused by anti-E antibody.

Objective : To pathological changes and inducible nitric oxide synthase(iNOS), phosphorylated insulin receptor substrate 1 serine 307(p-IRS1ser 307), phosphorylated protein kinase B serine 473(p-AKTser 473), glycogen synthase kinase-3β(GSK-3β), and gluconeogenic synthase(GS) proteins were observed in the liver of rats under the condition of chronic intermittent hypoxia-replicated oxygen in control. And to explore the role of iNOS/IRS1/AKT/GSK-3β signaling pathway in chronic intermittent hypoxia-induced insulin resistance. Methods : Forty SD rats were randomly divided into a control group(NC group) and an experimental group(CIH group), with 20 rats in each group. The NC group was placed in a normoxic environment for 12 weeks, while the CIH group was first subjected to intermittent hypoxia for 8 weeks, and then resumed normoxic rearing until the 12th week. Fasting blood glucose(FBG) and fasting insulin(FINS) were measured at baseline, week 8 and week 12, and liver tissues were taken for pathology and measurement of iNOS, p-IRS1ser 307, p-AKTser 473, GSK3β and GS levels, to compare the differences between groups. Results: t baseline, there was no significant difference in liver pathology between the two groups, and the observed indexes were not statistically significant(P>0.05); at 8 weeks, compared with the NC group, liver pathology in the CIH group showed significant disorganization of hepatic blood sinusoids and hepatocyte cords, obvious hepatocyte edema, smaller nuclei, increased lymphocyte infiltration, and a small number of fat vacuoles, significantly higher levels of FBG, FINS, insulin resistance index(HOMA-IR), iNOS mRNA, p-IRS1ser 307 protein, GSK-3β protein levels, and decreased p-AKTser 473 protein and GS protein levels, all of which were statistically significant(allP<0.05). IRS1ser 307 protein, GSK-3β protein levels were increased, p-AKTser 473 protein and GS protein levels were decreased, and the differences were statistically significant(allP<0.05); at 12 weeks, no lymphocyte infiltration was seen in the CIH group compared with that of the NC group and fat vacuoles significantly increased, and there was no improvement in the other pathological damage that had already occurred, and the levels of p-AKTser 473 protein significantly increased. AKTser 473 protein level significantly increased, p-IRS1ser 307 protein and GS protein levels were significantly reduced, all of which were statistically significant(allP<0.05), and the rest of the observational indexes were not statistically significant. Pearson′s correlation analysis showed that HOMA-IR of CIH group was significantly positively correlated with the levels of iNOS mRNA, p-IRS1ser 307 protein, and GSK-3β protein at 8 weeks(r=0.874, 0.817,0.872;allP<0.05), and significantly negatively correlated with the levels of p-AKTser 473 protein and GS protein(r=-0.886,-0.879;allP<0.05). Conclusion Chronic intermittent hypoxia can lead to hepatic pathological damage that cannot be reversed even by reoxygenation interventions and may mediate the development of insulin resistance by upregulating the IRS1/AKT/GSK-3β signaling pathway through the upregulation of iNOS mRNA expression.

OBJECTIVES: The application of photodynamic therapy in solid tumors has attracted increasing attention in recent years, and the efficiency of photosensitizers is a crucial determinant of therapeutic efficacy. This study aims to evaluate the cytotoxic effects of a novel photosensitizer, PEG-MTPABZ-PyC, in photodynamic therapy against gastric cancer cells. METHODS: Gastric cancer MKN45 cells were treated with PEG-MTPABZ-PyC. A high-content live-cell imaging system was used to assess the cellular uptake kinetics and subcellular localization of the photosensitizer. The cytotoxic effects of PEG-MTPABZ-PyC-mediated photodynamic therapy were examined using the cell counting kit-8 (CCK-8) assay and flow cytometry, while the intrinsic cytotoxicity of the photosensitizer alone was verified by the CCK-8 assay. Intracellular reactive oxygen species (ROS) generation after photodynamic therapy was detected using 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA). RESULTS: PEG-MTPABZ-PyC alone exhibited no cytotoxicity toward MKN45 cells, indicating excellent cytocompatibility. The compound efficiently entered cells within 6 hours and localized predominantly in lysosomes. Upon light irradiation, PEG-MTPABZ-PyC-mediated photodynamic therapy induced significant cytotoxicity compared with the control group (P<0.05) and generated abundant intracellular ROS. CONCLUSIONS The novel photosensitizer PEG-MTPABZ-PyC demonstrates potent photodynamic cytotoxicity against gastric cancer cells, showing promising potential for further development in gastric cancer photodynamic therapy.
Humans ; Stomach Neoplasms/drug therapy* ; Photochemotherapy/methods* ; Photosensitizing Agents/pharmacology* ; Cell Line, Tumor ; Polyethylene Glycols/chemistry* ; Reactive Oxygen Species/metabolism* ; Mesoporphyrins/pharmacology*

In drug clinical trials,the protection of subjects'privacy should take precedence over scientific exploration and societal benefits.This paper compared the differences in legal systems for the protection of subjects'privacy in China and abroad by employing methods such as literature review,case analysis,and comparison of domestic and foreign situations.It also elucidated the current ethical challenges faced by the protection of subjects'privacy,which primarily included imperfect laws and regulations,insufficient protection awareness among relevant personnel,risks of privacy breaches in critical links,and increased difficulty in data confidentiality under emerging technologies.From the perspective of responsible innovation,specific paths were proposed to protect subjects'privacy by using the four-dimensional structure of"prediction-reflection-negotiation-feedback,"maintaining a balance between ethical principles and technological innovation,ensuring the ethical and compliant nature of drug clinical trials,and thus better contributing to the cause of human health.

Objective:To explore the impact of the Yiqi Huoxue formula combined with nucleos(t)ide analogs (NAs) antiviral therapy on the serum N-glycan abundance in patients with hepatitis B-related liver fibrosis so as to clarify the application value of the oligosaccharide chain test (GT) for dynamic monitoring of the liver fibrosis progression.Methods:Sixty-two cases diagnosed with chronic hepatitis B at the Department of Hepatology, the Third Hospital of Hebei Medical University, between January 2020 and December 2022 were enrolled and divided into a Yiqi Huoxue Formula (YQHX) combined with NAs group and an NAs monotherapy group ( n=31), with 31 cases in each group for a total of 96 weeks of follow-up. Patient's basic clinical characteristics and liver stiffness measurement (LSM) were collected. GT was used simultaneously to detect the serum N-glycan profile and abundance changes. The nonparametric Mann-Whitney U test was used to compare the differences between the two groups. Enumeration data were expressed as number of cases and percentages (%). The χ2 test was used to compare constituent ratios between two or more groups. Correlation analysis was performed using the Spearman method, with P<0.05 considered statistically significant. Results:The proportion of patients was significantly higher in the YQHX combined with NAs group than the NAs monotherapy group [61.29% (19/31) vs. 9.68% (3/31), P<0.05] with no progression in liver fibrosis staging following 96 weeks of follow-up. The abundance of the N-glycan marker peak 8 [triantennary N-glycan (NA3)] had resulted in significant change for liver fibrosis improvements ( P<0.05), which predicts liver fibrosis progression and reversal in populations sensitive to traditional Chinese medicine. Conclusion:The combined application of Yiqi Huoxue formula and NAs can significantly promote the improvement rate of hepatitis B-related liver fibrosis. Serum N-glycan peak 8 may serve as a potential biomarker for monitoring the reversal of hepatitis B-related liver fibrosis.

BACKGROUND: This study aimed to investigate programmed death-ligand 1 tumor proportion score in predicting the safety and efficacy of PD-1/PD-L1 antibody-based therapy in treating patients with advanced non-small cell lung cancer (NSCLC) in a real-world setting. METHODS: This retrospective, multicenter, observational study enrolled adult patients who received PD-1/PD-L1 antibody-based therapy in China and met the following criteria: (1) had pathologically confirmed, unresectable stage III-IV NSCLC; (2) had a baseline PD-L1 tumor proportion score (TPS); and (3) had confirmed efficacy evaluation results after PD-1/PD-L1 treatment. Logistic regression, Kaplan-Meier analysis, and Cox regression were used to assess the progression-free survival (PFS), overall survival (OS), and immune-related adverse events (irAEs) as appropriate. RESULTS: A total of 409 patients, 65.0% ( n = 266) with a positive PD-L1 TPS (≥1%) and 32.8% ( n = 134) with PD-L1 TPS ≥50%, were included in this study. Cox regression confirmed that patients with a PD-L1 TPS ≥1% had significantly improved PFS (hazard ratio [HR] 0.747, 95% confidence interval [CI] 0.573-0.975, P = 0.032). A total of 160 (39.1%) patients experienced 206 irAEs, and 27 (6.6%) patients experienced 31 grade 3-5 irAEs. The organs most frequently associated with irAEs were the skin (52/409, 12.7%), thyroid (40/409, 9.8%), and lung (34/409, 8.3%). Multivariate logistic regression revealed that a PD-L1 TPS ≥1% (odds ratio [OR] 1.713, 95% CI 1.054-2.784, P = 0.030) was an independent risk factor for irAEs. Other risk factors for irAEs included pretreatment absolute lymphocyte count >2.5 × 10 9 /L (OR 3.772, 95% CI 1.377-10.329, P = 0.010) and pretreatment absolute eosinophil count >0.2 × 10 9 /L (OR 2.006, 95% CI 1.219-3.302, P = 0.006). Moreover, patients who developed irAEs demonstrated improved PFS (13.7 months vs. 8.4 months, P <0.001) and OS (28.0 months vs. 18.0 months, P = 0.007) compared with patients without irAEs. CONCLUSIONS A positive PD-L1 TPS (≥1%) was associated with improved PFS and an increased risk of irAEs in a real-world setting. The onset of irAEs was associated with improved PFS and OS in patients with advanced NSCLC receiving PD-1/PD-L1-based therapy.
Humans ; Carcinoma, Non-Small-Cell Lung/metabolism* ; Male ; Female ; Retrospective Studies ; Middle Aged ; Lung Neoplasms/metabolism* ; Aged ; B7-H1 Antigen/metabolism* ; Programmed Cell Death 1 Receptor/metabolism* ; Adult ; Aged, 80 and over ; Immune Checkpoint Inhibitors/therapeutic use*

Objective To develop and evaluate a quality assessment system for palliative care nursing homes that integrate medical and elderly care services based on the SERVQUAL model,thereby providing references for assessment and improvement of the quality of palliative care.Methods Using SERVQUAL model,a preliminary evaluation of a quality assessment system for palliative care was developed through literature review and semi-structured interviews.Two rounds of expert consultation with Delphi method were conducted to refine and finalise the assessment system.Results The two rounds of expert consultation each involved 15 questionnaires,all effectively returned(100.00%response rate),reflecting strong expert engagement.The expert authority coefficients were 0.78.The Kendall's W of primary irdicators were 0.198 in the first round to 0.235 in the second(P<0.05),showing improved consensus.The mean importance scores in the second consensus were 3.93-5.00,with coefficients of variation of 0-0.20,and weights of 0.01-0.28,respectively.The finallised assessment system comprised 5 primary indicators,17 secondary indicators and 64 tertiary indicators.Conclusion The SERVQUAL-based palliative care quality assessment system for integrated medical-elderly care nursing homes is scientifically rigorous and practically applicable.It offers a standardised tool for assessment and improvement of the quality of palliative care in nursing homes with integrated medical and elderly care services,promotes homogenisation of nursing quality and ultimately meets the needs of residents and their families.

Objective To develop and evaluate a quality assessment system for palliative care nursing homes that integrate medical and elderly care services based on the SERVQUAL model,thereby providing references for assessment and improvement of the quality of palliative care.Methods Using SERVQUAL model,a preliminary evaluation of a quality assessment system for palliative care was developed through literature review and semi-structured interviews.Two rounds of expert consultation with Delphi method were conducted to refine and finalise the assessment system.Results The two rounds of expert consultation each involved 15 questionnaires,all effectively returned(100.00%response rate),reflecting strong expert engagement.The expert authority coefficients were 0.78.The Kendall's W of primary irdicators were 0.198 in the first round to 0.235 in the second(P<0.05),showing improved consensus.The mean importance scores in the second consensus were 3.93-5.00,with coefficients of variation of 0-0.20,and weights of 0.01-0.28,respectively.The finallised assessment system comprised 5 primary indicators,17 secondary indicators and 64 tertiary indicators.Conclusion The SERVQUAL-based palliative care quality assessment system for integrated medical-elderly care nursing homes is scientifically rigorous and practically applicable.It offers a standardised tool for assessment and improvement of the quality of palliative care in nursing homes with integrated medical and elderly care services,promotes homogenisation of nursing quality and ultimately meets the needs of residents and their families.

In drug clinical trials,the protection of subjects'privacy should take precedence over scientific exploration and societal benefits.This paper compared the differences in legal systems for the protection of subjects'privacy in China and abroad by employing methods such as literature review,case analysis,and comparison of domestic and foreign situations.It also elucidated the current ethical challenges faced by the protection of subjects'privacy,which primarily included imperfect laws and regulations,insufficient protection awareness among relevant personnel,risks of privacy breaches in critical links,and increased difficulty in data confidentiality under emerging technologies.From the perspective of responsible innovation,specific paths were proposed to protect subjects'privacy by using the four-dimensional structure of"prediction-reflection-negotiation-feedback,"maintaining a balance between ethical principles and technological innovation,ensuring the ethical and compliant nature of drug clinical trials,and thus better contributing to the cause of human health.

Objective:To explore the impact of the Yiqi Huoxue formula combined with nucleos(t)ide analogs (NAs) antiviral therapy on the serum N-glycan abundance in patients with hepatitis B-related liver fibrosis so as to clarify the application value of the oligosaccharide chain test (GT) for dynamic monitoring of the liver fibrosis progression.Methods:Sixty-two cases diagnosed with chronic hepatitis B at the Department of Hepatology, the Third Hospital of Hebei Medical University, between January 2020 and December 2022 were enrolled and divided into a Yiqi Huoxue Formula (YQHX) combined with NAs group and an NAs monotherapy group ( n=31), with 31 cases in each group for a total of 96 weeks of follow-up. Patient's basic clinical characteristics and liver stiffness measurement (LSM) were collected. GT was used simultaneously to detect the serum N-glycan profile and abundance changes. The nonparametric Mann-Whitney U test was used to compare the differences between the two groups. Enumeration data were expressed as number of cases and percentages (%). The χ2 test was used to compare constituent ratios between two or more groups. Correlation analysis was performed using the Spearman method, with P<0.05 considered statistically significant. Results:The proportion of patients was significantly higher in the YQHX combined with NAs group than the NAs monotherapy group [61.29% (19/31) vs. 9.68% (3/31), P<0.05] with no progression in liver fibrosis staging following 96 weeks of follow-up. The abundance of the N-glycan marker peak 8 [triantennary N-glycan (NA3)] had resulted in significant change for liver fibrosis improvements ( P<0.05), which predicts liver fibrosis progression and reversal in populations sensitive to traditional Chinese medicine. Conclusion:The combined application of Yiqi Huoxue formula and NAs can significantly promote the improvement rate of hepatitis B-related liver fibrosis. Serum N-glycan peak 8 may serve as a potential biomarker for monitoring the reversal of hepatitis B-related liver fibrosis.