Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO₂), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO₂ maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.

Currently, the quality control of traditional Chinese medicine (TCM) encounters substantial technical challenges, necessitating the urgent development of innovative evaluation methods for key quality attributes. This study proposes the construction of a quality assurance system centered on key quality attributes, with systems biology and chemical biology serving as its core components: utilizing multi-omics technologies to systematically analyze the core functional modules of TCM, integrating pharmacological approaches to precisely identify key bioactive components; applying chemical biology to elucidate the interaction mechanisms between components and their targets, while screening for quality markers associated with specific functions and clinical indications; establishing an integrated quality control system based on quality markers and biological potency to ensure comprehensive quality management from raw herb materials through to finished products. Additionally, it emphasizes the formulation of compatibility rules grounded in TCM theory, integrates clinical practice and multi-dimensional value assessments, and establishes a novel quality evaluation model with distinct TCM characteristics.This offers a comprehensive and systematic approach to ensuring the safety and efficacy of TCM while facilitating the high-quality development of the TCM industry.

Objective To systematically evaluate the similarities between six-gene-modified pig red blood cells and human red blood cells in multiple dimensions, including cell morphology, hematological parameters, microbial safety and serological compatibility, and to verify the feasibility of using six-gene-modified pig red blood cells as an alternative for blood transfusion in cases of traumatic hemorrhage. Methods Blood samples from O-type human donor, six-gene-modified O-type pig and wild-type O-type pig were collected. The pig gene-modified phenotype was identified by flow cytometry. The morphology was observed using field emission scanning electron microscopy and the diameter was measured. Hematological tests, including blood routine, osmotic fragility test, adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) content and partial pressure of oxygen at 50% hemoglobin saturation (P50), were conducted to assess the deformability of red blood cells. Real-time fluorescent quantitative polymerase chain reaction was used to screen for zoonotic viruses, and cross-matching was performed using column gel method, condensation amine method and saline tube method. Results The six-gene-modified pig red blood cells successfully knocked out the α-Gal, Neu5Gc and Sda antigens and expressed hCD55. Wild-type pig red blood cells expressed these three antigens but did not express human complement regulatory proteins. All three types of red blood cells presented a biconcave discoid morphology. The diameter of O-type human red blood cells was larger than that of the six-gene-modified porcine red blood cells and wild-type pig red blood cells (all P < 0.05). The average red blood cell volume and average red blood cell hemoglobin of O-type humans were higher than those of the six-gene-modified pig red blood cells, and the distribution width-variable coefficient of O-type human red blood cells was lower than that of the six-gene-modified pig red blood cells (all P < 0.05). There were no statistically significant differences in the permeability fragility, ATP content, 2,3-DPG content, P50 and red blood cell deformability indicators of the three types of red blood cells (all P > 0.05). The screening for zoonotic viruses in human and animal cells of the six-gene-modified pigs and wild-type pigs was negative. The cross-matching results showed that in the column gel method and condensation amine method, the primary and secondary sides of the six-gene-modified pig red blood cells did not agglutinate with human red blood cells, while the wild-type pig red blood cells agglutinated with human red blood cells in all cross-matching methods. Conclusions The six-gene-modified pig red blood cells are highly similar to human red blood cells in terms of morphology, key hematological parameters, energy metabolism, oxygen-carrying capacity, mechanical properties, microbial safety and serological compatibility, providing an experimental basis for the clinical transformation of xenotransfusion.

This study was conducted retrospectively on a cohort of 68 patients with steroid 5 α-reductase 2 (SRD5A2) deficiency and 46,XY disorders of sex development (DSD). Whole-exon sequencing revealed 28 variants of SRD5A2 , and further analysis identified seven novel mutants. The preponderance of variants was observed in exon 1 and exon 4, specifically within the nicotinamide adenine dinucleotide phosphate (NADPH)-binding region. Among the entire cohort, 53 patients underwent initial surgery at Sichuan Provincial People's Hospital (Chengdu, China). The external genitalia scores (EGS) of these participants varied from 2.0 to 11.0, with a mean of 6.8 (standard deviation [s.d.]: 2.5). Thirty patients consented to hormone testing. Their average testosterone-to-dihydrotestosterone (T/DHT) ratio was 49.3 (s.d.: 23.4). Genetic testing identified four patients with EGS scores between 6 and 9 as having this syndrome; and their T/DHT ratios were below the diagnostic threshold. Furthermore, assessments conducted using the crystal structure of human SRD5A2 have provided insights into the potential pathogenic mechanisms of these novel variants. These mechanisms include interference with NADPH binding (c.356G>C, c.365A>G, c.492C>G, and c.662T>G) and destabilization of the protein structure (c.727C>T). The c.446-1G>T and c.380delG variants were verified to result in large alterations in the transcripts. Seven novel variations were identified, and the variant database for the SRD5A2 gene was expanded. These findings contribute to the progress of diagnostic and therapeutic approaches for individuals with SRD5A2 deficiency.
Humans ; 3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Disorder of Sex Development, 46,XY/blood* ; Male ; Membrane Proteins/genetics* ; Child, Preschool ; Child ; Retrospective Studies ; Adolescent ; Female ; Mutation ; Testosterone/blood* ; Infant ; Dihydrotestosterone/blood*

OBJECTIVE: To investigate the relationship between peripheral blood T-cell immunoglobulin mucin-3 (TIM-3) and iron overload in patients with myelodysplastic syndrome (MDS) undergoing red blood cell transfusion. METHODS: 120 MDS patients who received treatment at Wuhan Third Hospital from June 2020 to May 2022 were included and analyzed as research subjects, all of whom met the indications for red blood cell transfusion. Blood routine and biochemical indicators were tested before transfusion, and general clinical data of the patients were statistically analyzed. The iron metabolism status of the patients were evaluated. The clinical characteristics of patients with iron overload and the factors affecting iron overload were analyzed. And a correlation analysis was conducted between TIM-3 and other factors affecting iron overload. RESULTS: Among the 120 MDS patients included in this study, 82 cases (68.33%) were detected to have iron overload after red blood cell transfusion. The occurrence time of iron overload was 20-42 weeks, with an average time of 32.35±5.26 weeks, calculated from the first transfusion of red blood cells. The proportion of patients with high-risk and extremely high-risk according to the revised International Prognostic Scoring System (IPSS-R) and WHO classification-based Prognostic Scoring System (WPSS), the volume of blood transfusions, the proportion of transfusion-dependent patients, and the levels of serum hepcidin (Hepc), erythropoietin (EPO), and TIM-3 in patients with iron overload were higher than those in patients with normal iron metabolism, and the differences were statistically significant (P < 0.05). Logistic regression analysis showed that high-risk and extremely high-risk according to WPSS, blood transfusion volume, transfusion dependence, and upregulation of serum Hepc, EPO, and TIM-3 expression were factors affecting iron overload in MDS patients undergoing red blood cell transfusion (P < 0.05). Pearson correlation analysis showed that serum TIM-3 level in MDS patients were positively correlated with the other factors affecting iron overload (P < 0.05). CONCLUSION Serum TIM-3 is associated with iron overload in MDS patients undergoing red blood cell transfusion, and upregulation of serum TIM-3 expression increases the risk of iron overload after red blood cell transfusion.
Humans ; Myelodysplastic Syndromes/blood* ; Iron Overload/blood* ; Hepatitis A Virus Cellular Receptor 2/blood* ; Erythrocyte Transfusion ; Male ; Female ; Middle Aged ; Aged ; Iron

Objective: Hundreds of countries have implemented lockdown policies to slow the spread of coronavirus disease-2019 (COVID-19), but the impact of these measures on maternal mental health is not well understood. Methods: This study integrated a stress-process model to examine the pathways from lockdown-related stressors to prenatal psychological outcomes, with COVID-19 coping strategies (COP) and self-efficacy in managing negative affect (NEG) as mediators and lockdown duration, hours on pandemic-related information, and number of pregnancies as moderators. Pregnant women in Shanghai completed the Regulatory Emotional Self-Efficacy Scale, COVID-19 Coping Scale, Depression, Anxiety, and Stress Scale-21. Structural equation modeling (SEM) was used to test and modify the hypothetical model, and moderated mediation and slope analyses were undertaken. Results: In the final SEM demonstrating satisfactory fit, three stressors—decreased household income, insufficient daily supplies, and acquired infections—showed positive direct relationships with NEG and COP. Acquired infections, NEG, and COP were identified as direct predictors of mental health outcomes. The relationship between these three stressors and mental health was mediated by NEG and COP. Additionally, the number of pregnancies moderated the mediating effect of COP; this effect was more pronounced among first-time pregnant women than those with multiple pregnancies. Conclusion This study provides insights into how lockdown measures impact psychological outcomes in pregnant women quarantined at home. Interventions aimed at increasing coping strategies may be more effective for primiparous women during future public health emergencies.

Objective: Hundreds of countries have implemented lockdown policies to slow the spread of coronavirus disease-2019 (COVID-19), but the impact of these measures on maternal mental health is not well understood. Methods: This study integrated a stress-process model to examine the pathways from lockdown-related stressors to prenatal psychological outcomes, with COVID-19 coping strategies (COP) and self-efficacy in managing negative affect (NEG) as mediators and lockdown duration, hours on pandemic-related information, and number of pregnancies as moderators. Pregnant women in Shanghai completed the Regulatory Emotional Self-Efficacy Scale, COVID-19 Coping Scale, Depression, Anxiety, and Stress Scale-21. Structural equation modeling (SEM) was used to test and modify the hypothetical model, and moderated mediation and slope analyses were undertaken. Results: In the final SEM demonstrating satisfactory fit, three stressors—decreased household income, insufficient daily supplies, and acquired infections—showed positive direct relationships with NEG and COP. Acquired infections, NEG, and COP were identified as direct predictors of mental health outcomes. The relationship between these three stressors and mental health was mediated by NEG and COP. Additionally, the number of pregnancies moderated the mediating effect of COP; this effect was more pronounced among first-time pregnant women than those with multiple pregnancies. Conclusion This study provides insights into how lockdown measures impact psychological outcomes in pregnant women quarantined at home. Interventions aimed at increasing coping strategies may be more effective for primiparous women during future public health emergencies.

Objective: Hundreds of countries have implemented lockdown policies to slow the spread of coronavirus disease-2019 (COVID-19), but the impact of these measures on maternal mental health is not well understood. Methods: This study integrated a stress-process model to examine the pathways from lockdown-related stressors to prenatal psychological outcomes, with COVID-19 coping strategies (COP) and self-efficacy in managing negative affect (NEG) as mediators and lockdown duration, hours on pandemic-related information, and number of pregnancies as moderators. Pregnant women in Shanghai completed the Regulatory Emotional Self-Efficacy Scale, COVID-19 Coping Scale, Depression, Anxiety, and Stress Scale-21. Structural equation modeling (SEM) was used to test and modify the hypothetical model, and moderated mediation and slope analyses were undertaken. Results: In the final SEM demonstrating satisfactory fit, three stressors—decreased household income, insufficient daily supplies, and acquired infections—showed positive direct relationships with NEG and COP. Acquired infections, NEG, and COP were identified as direct predictors of mental health outcomes. The relationship between these three stressors and mental health was mediated by NEG and COP. Additionally, the number of pregnancies moderated the mediating effect of COP; this effect was more pronounced among first-time pregnant women than those with multiple pregnancies. Conclusion This study provides insights into how lockdown measures impact psychological outcomes in pregnant women quarantined at home. Interventions aimed at increasing coping strategies may be more effective for primiparous women during future public health emergencies.

Objective: Hundreds of countries have implemented lockdown policies to slow the spread of coronavirus disease-2019 (COVID-19), but the impact of these measures on maternal mental health is not well understood. Methods: This study integrated a stress-process model to examine the pathways from lockdown-related stressors to prenatal psychological outcomes, with COVID-19 coping strategies (COP) and self-efficacy in managing negative affect (NEG) as mediators and lockdown duration, hours on pandemic-related information, and number of pregnancies as moderators. Pregnant women in Shanghai completed the Regulatory Emotional Self-Efficacy Scale, COVID-19 Coping Scale, Depression, Anxiety, and Stress Scale-21. Structural equation modeling (SEM) was used to test and modify the hypothetical model, and moderated mediation and slope analyses were undertaken. Results: In the final SEM demonstrating satisfactory fit, three stressors—decreased household income, insufficient daily supplies, and acquired infections—showed positive direct relationships with NEG and COP. Acquired infections, NEG, and COP were identified as direct predictors of mental health outcomes. The relationship between these three stressors and mental health was mediated by NEG and COP. Additionally, the number of pregnancies moderated the mediating effect of COP; this effect was more pronounced among first-time pregnant women than those with multiple pregnancies. Conclusion This study provides insights into how lockdown measures impact psychological outcomes in pregnant women quarantined at home. Interventions aimed at increasing coping strategies may be more effective for primiparous women during future public health emergencies.

In analytical techniques such as ion mobility spectrometry(IMS)and mass spectrometry(MS),the ionization efficiency of target analytes is primarily constrained by the type of ionization source and factors such as the species and number density of the reactant ions.Systematic investigation into the reactivity differences of various reactant ions under varying conditions can not only significantly enhance the detection sensitivity of target compound product ions but also provide a theoretical foundation for establishing efficient detection methods based on ion-molecule reaction mechanisms.In this study,the pressure of a pressure-tunable photoionization tandem ion mobility spectrometry(PI-tandem-IMS)was reduced from ambient pressure(100 kPa)to low pressure(20 kPa)to systematically examine the reactivity differences between two negative reactant ions,CO3-and CO4-,and methyl salicylate(MeSA)under varying pressures.When the pressure decreased,the increased relative signal intensity of CO4-significantly influenced the detection sensitivity of the characteristic product ion[MeSA·O2]-.Based on differences in ion mobility(k0),the delay time for the opening of TPG2 was adjusted to selectively inject CO-3 and CO-4 in the drift region 2.Independent characterization of the reactivity of these reactant ions with MeSA in the reaction region confirmed that CO4-exhibited superior reactivity toward MeSA.The theoretical model revealed an Arrhenius plot for the ion-molecule reaction between CO4-and MeSA,showing a positive correlation between the reaction rate coefficient(k)and temperature,the activation energy Ea was 62.45 kJ/mol.Furthermore,controlling parameters such as pressure or temperature significantly influenced the progression of this ion-molecule reaction,demonstrating the technical advantages of PI-tandem-IMS in mechanistic studies and regulation of ion-molecule reactions.