OBJECTIVE To investigate the effects and mechanisms of Lycium ruthenicum Murr. in improving sleep. METHODS Network pharmacology was employed to identify the active components of L. ruthenicum and their associated disease targets， followed by enrichment analysis. A caffeine‑induced zebrafish model of sleep deprivation was established ， and the zebrafish were treated with L. ruthenicum Murr. extract （LRME） at concentrations of 0.1， 0.2 and 0.4 mg/mL， respectively； 24 h later， behavioral changes of zebrafish and pathological alterations in brain neurons were subsequently observed. The levels of inflammatory factors [interleukin-6 （IL-6）， IL-1β， IL-10， tumor necrosis factor-α （TNF-α）]， oxidative stress markers [superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， catalase （CAT）]， and neurotransmitters [5- hydroxytryptamine （5-HT）， γ-aminobutyric acid （GABA）， glutamic acid （Glu）， dopamine （DA）， and norepinephrine （NE）] were measured. The protein expression levels of protein kinase B1 （AKT1）， phosphorylated AKT1 （p-AKT1）， epidermal growth factor receptor （EGFR）， B-cell lymphoma 2 （Bcl-2）， sarcoma proto-oncogene，non-receptor tyrosine kinase （SRC）， and heat shock protein 90α family class A member 1 （HSP90AA1） in the zebrafish were also determined. RESULTS A total of 12 active components and 176 intersecting disease targets were identified through network pharmacology analysis. Among these， apigenin， naringenin and others were recognized as core active compounds， while AKT1， EGFR and others served as key targets； EGFR tyrosine kinase inhibitor resistance signaling pathway was identified as the critical pathway. The sleep improvement rates in zebrafish of LRME low-， medium-， and high-dose groups were 54.60%， 69.03% and 77.97%， 开发。E-mail：hjp_yft@163.com respectively， while the inhibition ratios of locomotor distance were 0.57， 0.83 and 0.95， respectively. Compared with the model group， the number of resting counts， resting time and resting distance were significantly increased/extended in LRME medium- and high-dose groups （P＜0.05）. Neuronal damage in the brain was alleviated. Additionally， the levels of IL-6， IL-1β， TNF-α， MDA， Glu， DA and NE， as well as the protein expression levels of AKT1， p-AKT1， EGFR， SRC and HSP90AA1， were markedly reduced （P＜0.05）， while the levels of IL-10， SOD， GSH-Px， CAT， 5-HT and GABA， as well as Bcl-2 protein expression， were significantly elevated （P＜0.05）. CONCLUSIONS L. ruthenicum Murr. demonstrates sleep-improving effects， and its specific mechanism may be related to the regulation of inflammatory responses， oxidative stress， neurotransmitter balance， and the EGFR tyrosine kinase inhibitor resistance signaling pathway.

Background: Influenza A virus (IAV) is a negative-sense RNA virus of the Orthomyxoviridae family and is the etiological agent of a highly contagious acute respiratory disease that can lead to acute lung injury. Objective: To elucidate the molecular mechanisms of IAV infection, an integrative research approach combining gene expression profiling, multinetwork analysis, and in vivo experimental validations was employed. Methods: First, a series of network-based analyses were performed, including protein-protein interaction network construction, weighted gene co-expression network analysis, and subsequent gene set enrichment analysis, to identify the major underlying mechanisms of IAV infection. Following gene expression analysis, core targets, both direct and indirect regulators, were screened. An IAV (H1N1) strain A/PR/8/34-induced acute lung injury mouse model was constructed for in vivo validations. Batch one included two groups to evaluate findings from the multi-network analysis: Mock (n = 10; 5 males and 5 females) and IAV (n = 10; 5 males and 5 females). Batch two included three groups to assess the role of toll-like receptor 3 (TLR3) in IAV infection: Mock (n = 6; 3 males and 3 females), IAV (n = 6; 3 males and 3 females), and TLR3 inhibitor (n = 6; 3 males and 3 females). Body weight was measured on days 0, 3, and 5 after infection. On day 5, lung tissues were collected to assess viral load and histopathological changes. Key targets were examined using enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining, both in sera and lung tissues. Results: IAV infection was significantly associated with dysregulation of the immune-inflammation system, such as the LTR, nucle-otide-binding oligomerization domain-(NOD) like receptor, retinoic acid-inducible gene I-like receptor, and nuclear factor kappa-B signaling pathways. Gene set enrichment analysis further indicated that the TLR and necroptosis signaling pathways played crucial roles in the progression of IAV infection (TLR signaling pathway normalized enrichment score = 2.3941, P = 1.00 × 10 ⁻¹⁰; necroptosis normalized enrichment score = 1.9421, P = 6.21 × 10 ⁻⁷). Among the core targets, TLR3 and mixed lineage kinase domain-like protein (MLKL) may regulate gene expression at the transcriptional level (all P < 0.05). In vivo validation using an IAV (PR8) infected acute lung injury mouse model demonstrated increased viral load and lung index, alveolar structural damage, and inflammatory cell infiltration. Immunofluorescence staining exhibited large gaps in Lamin B1 staining and breaches in Emerin signals following IAV-PR8 infection. Expression levels of TLR3, p-receptor-interacting serine/threonine-protein kinase 3 (RIPK3)/RIPK3, and p-mixed lineage kinase domain-like protein (MLKL)/MLKL proteins in lung tissues, as well as proinflammatory factors and mediators in sera, were significantly elevated after IAV infection. Moreover, enhanced neutrophil infiltration (myeloperoxidase) and citrullinated histone H3 (a neutrophil extracellular trap-specific marker), both established indicators of neutrophil extracellular trap formation, were observed. Notably, treatment with a TLR3 inhibitor significantly ameliorated IAV-induced acute lung injury by regulating necroptosis-related targets. Conclusion: Our study provides network-based in vivo evidence that TLR3-receptor-interacting serine/threonine-protein kinase 3-MLKL-mediated necroptosis may underlie IAV-induced acute lung injury and could serve as a potential therapeutic target in severe influenza cases.

[Objective] To explore the effects of different methods on antibody detection through investigating the causes of cross-matching incompatible in a patient with gastric malignant tumor, and to establish flow cytometry protocol for confirming hemolytic transfusion reaction (HTR). [Methods] Antibodies in the patient's serum were identified by red blood cells (RBCs) blood grouping, antibody screening and identification, acid elution test and PEG enhancement test. To confirm HTR, patient RBCs, proximal and distal ends RBCs, separated by capillary centrifugation, were tested by direct antiglobulin test (DAT) and Jk^a antigen single label and double label flow cytometry. [Results] Routine serological technology revealed the presence of anti-C, e (titer:2) and anti-Jka (titer >1) in the patient’s serum. After separation using capillary centrifugation technology, both the proximal and distal DAT and Jk^a antigen tests were negative. Both DAT and Jk^a antigen positive red blood cells (0.21%, 6/6 327) were found in the patient's blood samples by flow cytometry. After separation of blood samples by capillary centrifugation, there were significantly more DAT and Jk^a antigen double-positive RBCs in the distal end (0.43%, 33/7 707) than in the proximal end (0.09%, 15/7 225). Two blood samples were screened from over 100 donor blood samples that are compatible with the patient's cross-matching, and the transfusion effect was favorable. [Conclusion] Serological methods combined with flow cytometry could improve the sensitivity of antibody detection, provide a more accurate basis for the diagnosis of HTRs, and guarantee the safety of blood transfusion.

Objective Based on multi-objective optimization,a design method for Voronoi bionic porous scaffolds tailored to different degrees of bone defects was proposed.Methods First,the effects of design parameters on mechanical and biological properties of the scaffolds were investigated.The response surface models were then established respectively for the design parameters and performance indicators(specific surface area,elastic modulus,yield strength,and permeability).Using a cubic scaffold with side length of 15 mm as an example(assuming a corresponding bone defect of the same dimension),multi-objective optimization of the scaffold was finally conducted using the non-dominated genetic algorithm-Ⅱ algorithm,while considering the elastic modulus and permeability ranges of bone tissues as performance constraints.Results The degree of anisotropy in Voronoi scaffolds was influenced by the number of seed points,while the size and scaling factors of the scaffolds exclusively impacted the rod diameter and rod length.Using the design method of this study,the optimal scaffold with specific defect size satisfying mechanical and biological properties was designed.The optimal scaffold meeting different strength requirements was designed by adjusting the yield strength to change the utopia point.Conclusions A design method for Voronoi bionic porous scaffolds based on multi-objective optimization is proposed.This method can be applied to bone defects at varying degrees and provides a new idea for the personalized design of bone tissue engineering scaffolds.

Objective: To analyze the results of national external quality assessment (EQA) for transfusion compatibility test from 2018 to 2023, with the aim of providing references for improving laboratory testing quality and ensuring the safety of clinical blood transfusion. Methods: Three EQA programs were conducted annually, each distributing 22 quality assessment samples. Participating transfusion laboratories were required to complete testing within specified deadlines and to submit results along with documentation of testing methodologies, reagents, and equipment used. National Center for Clinical Laboratories (NCCL) conducted statistical analysis of laboratory results, evaluated testing outcomes and related circumstances, and provided feedback to participating laboratories. EQA data from transfusion laboratories across China from 2018 to 2023 were collected and systematically analyzed. Results: From 2018 to 2023, the qualification rates for all five items (ABO forward typing, ABO reverse typing, Rh blood group typing, antibody screening, and cross-matching) were 67.59%, 77.11%, 77.38%, 72.78%, 79.96%, and 85.16%, respectively. The mean qualification rates for ABO forward typing, ABO reverse typing, RhD blood group typing, antibody screening, and cross-matching over the past six years were 96.25%±0.59%, 90.45%±4.52%, 96.05%±0.71%, 90.88%±2.86%, and 88.34%±3.48%, respectively. The qualification rates in 2019, 2020, 2022, and 2023 all showed a stable trend of "blood stations>tertiary hospitals>secondary hospitals". The mean qualification rate of laboratories in secondary hospitals from 2018 to 2023 was significantly lower than those of laboratories in tertiary hospitals and blood stations (P<0.05), while no significant difference was observed between laboratories in tertiary hospitals and blood stations (P>0.05). The micro column agglutination method was the most widely used in all five tests. In the four test items, namely ABO forward typing, ABO reverse typing, antibody screening, and cross-matching, there was a statistically significant difference in the qualification rate of micro column agglutination method compared to other methods (P<0.05). There was a statistical difference in the qualification rate between manual and automated detection using micro column agglutination method in the cross-matching tests (P<0.05), whereas no significant difference was noted for the other test items (P>0.05). Conclusion: From 2018 to 2023, the number of laboratories participating in EQA activities has been increasing year by year, and the qualification rate has shown an overall upward trend. The type of laboratory is a key factor affecting the qualification rate, and the testing capabilities of some laboratories still need to be improved. The micro column agglutination method is widely used in transfusion compatibility tests. The established EQA program effectively monitors quality issues in laboratories, drives continuous improvement, and ensures sustained enhancement of testing standards to safeguard clinical blood safety.

Objective To investigate the situation of unconformity of positive and reverse ABO blood typ-ing between outpatient and inpatient patients in the hospital,and explore the reasons for unconformity of posi-tive and reverse ABO blood typing detected by microcolumn gel method.Methods A retrospective analysis was conducted on the medical records of totally 425 patients with ABO blood type mismatch who underwent ORTHO VISION Max testing in the hospital from August 2023 to December 2024.Comprehensive analysis was conducted using various methods including the saline tube method,irregular antibody screening,increas-ing plasma volume,4 ℃ enhancement test,absorption elution,and gene sequencing,etc.Results Among 128 192 cases of ABO blood type tests,425 cases showed unconformity of positive and reverse ABO blood typ-ing,accounting for 0.33％.The causes were as follows:antibody weakening in 316 cases(74.35％),bone mar-row transplantation in 40 cases(9.41％),interference from irregular antibodies in 34 cases(8.00％),antigen weakening in 13 cases(3.06％),subgroups in 10 cases(2.35％),cold agglutinins in 6 cases(1.41％),inter-ference from irregular and autoantibodies in 4 cases(0.94％),auto-sensitization of red blood cells in 1 case(0.24％),and other reasons in 1 case(0.24％).Among 316 cases with antibody weakening,239 cases(75.63％)involved type A(anti-B weakening),61 cases(19.30％)involved type B(anti-A weakening),8 ca-ses(2.53％)involved type O(anti-A weakening),6 cases(1.91％)involved type O(anti-B weakening),and 2 cases(0.63％)involved type O(both anti-A and anti-B weakening).In patients with antibody weakening,the disease distribution was mainly cardiovascular disease in 127 cases(40.19％),a history of tumors in 67 ca-ses(21.20％),kidney diseases in 11 cases(3.48％),and orthopedic diseases in 10 cases(3.16％).Conclusion The main reason for the unconformity of positive and reverse ABO blood typing in the hospital is the weakening of the antibodies,especially the predominance of type A(anti-B weakening).Weakened anti-bodies are predominantly in the distribution of cardiovascular disease.For subtype-induced disturbances,a comprehensive analysis combining serology and gene sequencing is recommended.For disease-induced uncon-formity of positive and reverse ABO blood typing,monitoring the patients should be strengthened.

Objective:To analyze the results of national external quality assessment (EQA) for transfusion compatibility test in 2023, and provide reference for quality management of clinical transfusion compatibility testing.Methods:The EQA of clinical transfusion compatibility testing by NCCL was performed 3 times in 2023 among included laboratories. The panel consisting of 22 samples was distributed to 4 186 laboratories across 31 provinces (Including 2 961 tertiary hospital laboratories, 1 085 secondary hospital laboratories, 23 primary hospital laboratories, 106 blood station laboratories and 11 independent clinical laboratories). Each panel contains 11 red blood cell and 11 plasma samples per 1.5 ml/tube. Each participant laboratory of the EQA program was required to carry out the detection and return results in expected time. Statistical analysis and evaluation on the reported results were conducted by NCCL from the aspects of regional distribution, laboratory grading, testing methodology, reagent and testing system usage.Results:The qualification rates of EQA for five items including ABO positive typing, ABO reverse typing, RhD blood type, antibody screening, and cross matching were 96.68%, 95.10%, 96.46%, 95.32%, and 91.04%, respectively. The EQA qualification rate of tertiary hospital laboratories was 87.77% (2 599/2 961), which was significantly higher than the 77.79% (844/1 085) of secondary hospital laboratories. There were significant differences in the qualification rate of participating laboratories among different regions. The utilization rates of micro column agglutination method in ABO positive typing, ABO reverse typing, RhD blood type, antibody screening, and cross matching were 80.81% (10 080/12 474), 75.06% (9 337/12 440), 81.38% (10 118/12 433), 89.59% (11 104/12 394) and 76.25% (9 495/12 453), respectively. The qualification rate of micro column agglutination method was significantly higher than that of saline slide method in ABO positive typing detection ( P<0.05). The qualification rate of micro column agglutination method was significantly higher than that of the polyamine method and anti-human globulin test tube method in antibody screening ( P<0.05). There were statistically significant differences in qualification rate of 7 reagents in ABO reverse typing, antibody screening and cross matching ( P<0.05). There was no statistically significant difference in the qualification rate between the two detection systems for other reagents, except for the ABO reverse typing where the qualification rate of reagent 1 in a single system was higher than that in a mixed system ( P<0.05). Conclusion:The testing capabilities of clinical laboratories in different regions and different type varied significantly in China. Micro column agglutination method was the most popular selection in transfusion compatibility testing. The regents used in these laboratories showed good performance. However, the detection efficiency of some reagents still need to be improved. EQA could be used to evaluate, monitor, and improve the quality of testing.

Objective:To investigate the clinical application effects of free transplantation of lobulated inguinal flaps.Methods:This study was a retrospective observational study. From July 2019 to April 2024, 34 patients with skin defect wounds whose wounds in one part met the inclusion criteria were admitted to Tongren Hospital of Wuhan University & Wuhan Third Hospital, including 28 males and 6 females, aged 26 to 59 years. The wound area in the recipient area ranged from 3.0 cm×2.0 cm to 25.0 cm×20.0 cm. The lobulated inguinal flap pedicled with the branch of the superficial circumflex iliac artery were obtained in 19 patients, and the lobulated inguinal flap pedicled with the main artery of the superficial circumflex iliac artery and the superficial inferior epigastric artery were obtained in 15 patients. The total area of the flaps ranged from 6.0 cm×2.2 cm to 27.0 cm×23.0 cm. The flaps were divided into 2 to 4 lobes, and the area of each lobe ranged from 2.0 cm×1.0 cm to 17.0 cm×12.0 cm. Each lobe of the flaps was reassembled, spliced, or directly transplanted onto the wounds, and the donor wounds were sutured in layers. The survival of each lobe of the flaps and wound healing in the recipient and donor areas were observed, and the wound recovery in the recipient and donor areas were followed up. At the last follow-up, the patient's satisfaction with the efficacy was assessed by 5-grade Likert scale.Results:A small amount of necrosis appeared in the tip of one lobe of the flaps in 4 patients after surgery, which healed after trimming. The flaps of the remaining 30 patients survived. The wounds in the recipient areas healed smoothly. There was a small amount of necrosis at the suture edge of the donor areas in 3 patients, which healed after local trimming and dressing change. The donor wounds healed well in the remaining 31 patients. During the follow-up of 6 to 42 months, all the recipient wounds were well repaired, and the shape of the donor areas was good. At the last follow-up, 15 patients were very satisfied with the efficacy, 15 were relatively satisfied, and 4 were generally satisfied.Conclusions:Through preoperative ultrasonic examination and positioning, the inguinal flap is designed according to the course of blood vessels and lobulated with the branch of the superficial circumflex iliac artery or the main artery of the superficial circumflex iliac artery and the superficial inferior epigastric artery as the pedicles. The anatomical process is reliable and the blood flow of the flap after being lobulated is rich, which can meet the repair needs of various skin defect wounds. The repair effect is good, and the damage in the donor area is small, which is worthy of promotion.

Objective:To explore the efficacy of free medial sural artery perforator flap transplantation in repairing electrical burn wounds on hands and feet.Methods:This study was a retrospective observational study. From November 2017 to September 2023, 21 male patients aged 28-51 years with electrical burns on hands and feet who met the inclusion criteria were admitted to Tongren Hospital of Wuhan University & Wuhan Third Hospital. There were 23 wounds, of which 14 were on the hand and 9 on the foot; 9 were associated with exposed tendon injury, 6 were associated with bone exposure or necrosis, and 8 were associated with joint injury. The wound area after debridement was 4.0 cm×2.5 cm-14.0 cm×10.0 cm. For 2 relatively wide wounds and 2 adjacent fingers/toes wounds, the lobulated flaps centered on 2 medial sural artery perforators were designed and incised for repair. For other wounds, medial sural artery perforator flaps were designed and incised. The flap area was 5.0 cm×3.0 cm-16.0 cm×11.0 cm. The arteriovenous vessels of flap were anastomosed end-to-end with the arteriovenous vessels of the recipient area; the cutaneous nerves of 10 flaps were anastomosed with the nerves in hand wound, and the sural nerve bundle was cut to repair one digital nerve defect. The donor site wound was closed with tension-relieving sutures. Postoperative flap survival and wound healing at donor site were recorded. During follow-up, subsequent flap revision was recorded, the texture and appearance of the flap, as well as the scarring and functional recovery of the donor area of the lower leg, were observed. At the last follow-up, the recovery of hand flap sensation was observed, the satisfaction of patients with the treatment effect of each operation was investigated by using Likert scale, the hand function of the affected hand in patients with hand wounds was evaluated by using the trial standards for evaluation of partial function of upper extremity by the Hand Surgery Society of Chinese Medical Association, and the weight-bearing walking ability of the affected foot in patients with foot wounds was evaluated by Holden walking ability grading.Results:There was a slight necrosis at the distal end of one flap after surgery, which healed after dressing change. All 23 flaps survived. The sutures of the two donor areas were poorly healed due to high local tension, and the second sutures were performed after debridement and drainage, and the healing was good. The wounds of the remaining 21 donor sites healed well. Follow-up of 6-26 months after surgery showed that 3 flaps were slightly bloated, and the appearance was improved after flap reconstruction; the other flaps did not undergo subsequent revision. All flaps were soft and similar to the surrounding tissue morphology. Linear scar remained in the donor site of the lower leg, and walking function was normal. At the last follow-up, the protective sensation of the hand flap was restored; the patients were very satisfied with the results of 21 surgeries and were relatively satisfied with the results of 2 surgeries. Among the 14 patients with hand wounds, the affected hand function was rated as excellent in 10 cases, very good in 3 cases, and acceptable in one case, and the weight-bearing walking ability of the affected foot in 7 patients with foot wounds was all rated as grade Ⅴ.Conclusions:The medial sural artery perforator flap has the advantages of reliable blood supply, appropriate thickness and smoothness, and can be lobulated or cut according to the shape of the wound. The flap demonstrates superior aesthetic and functional restoration in repairing electrical burn wounds on hands and feet, achieving high patient satisfaction with the surgical treatment effect.

Objective:To investigate the effect of transforming growth factor beta 1 (TGF-β1) on the biological activity of infantile hemangioma (IH) -derived endothelial cells (HemECs) .Methods:Three proliferating IH tissues and three involuting IH tissues were collected from IH patients receiving surgical resection at the Department of Pediatric Surgery, West China Hospital, Sichuan University from February to August 2021. Primary HemECs were isolated from proliferating IH tissues, and human umbilical vein endothelial cells (HUVECs) served as the control. The TGF-β1 expression levels in tissues and cells were detected by immunohistochemical study and Western blot analysis. Cell counting kit-8 (CCK8) assay was performed to assess the effect of 0 (control group) - 100 ng/ml TGF-β1 on HemEC proliferation. HemECs were treated with 5 ng/ml TGF-β1 or without (control group), and after several hours of treatment, Transwell assay was performed to evaluate cell migration ability, and immunofluorescence assay to assess the changes in the expression of endothelial markers (platelet-endothelial cell adhesion molecule-1 [CD31], vascular endothelial cadherin [VE-cadherin]) and mesenchymal markers (α-smooth muscle actin [α-SMA], collagen type Ⅰ α 1 [COL1A1]). Comparisons between groups were conducted by t test or one-way analysis of variance. Results:Immunohistochemical study showed that proliferating IH tissues were stained positively for TGF-β1, which was expressed relatively abundantly; the percentages of TGF-β1-positive signal area were higher in the proliferating IH tissues (24.68% ± 3.74%) than in the involuting IH tissues (almost no expression). Western blot analysis revealed that the relative expression level of TGF-β1 was significantly higher in HemECs (1.08 ± 0.13) than in HUVECs (0.30 ± 0.04, t = 9.93, P < 0.001). CCK8 assay showed increased proliferative activity of HemECs in the 3.125-, 6.25-, 12.5-, 25-, 50- and 75-ng/ml TGF-β1 groups compared with the control group (all P < 0.05), and no significant difference was found between the 100-ng/ml TGF-β1 group and the control group ( P > 0.05). Transwell assay revealed an increased number of migratory HemECs in the 5-ng/ml TGF-β1 group (127 ± 6) compared with the control group (103 ± 9; t = 5.32, P < 0.01). Immunofluorescence assay showed significantly decreased fluorescence intensity of endothelial markers CD31 and VE-cadherin in the 5-ng/ml TGF-β1 group (5.441 ± 1.254, 5.073 ± 0.412, respectively) compared with the control group (9.518 ± 1.728,7.671 ± 0.921, t = 3.31, 4.46, P = 0.030, 0.011, respectively), and significantly increased fluorescence intensity of mesenchymal markers α-SMA and COL1A1 in the 5-ng/ml TGF-β1 group (8.074 ± 0.846, 5.885 ± 0.216, respectively) compared with the control group (0.393 ± 0.342, 0.295 ± 0.125, t = 14.58, 38.76, P < 0.001, < 0.000 1, respectively) . Conclusion:TGF-β1 was relatively highly expressed in the proliferating IH tissues and HemECs, and could promote the proliferation, migration and endothelial-to-mesenchymal transition of HemECs.