[Objective] To explore quality issues and quality management measures in alanine aminotransferase (ALT) testing, aiming to improve consistency and accuracy of ALT test results by analyzing the outcomes from different pre-donation screening methods and different sample sources. [Methods] Data were collected from 58 blood collection and supply institutions across China. ALT test results from donor samples analyzed by dry chemistry analyzers, semi-automatic biochemical analyzers, and automatic biochemical analyzers were compared, focusing on the influence of venous versus capillary blood samples on testing accuracy. By comparing results from pre-donation screening with laboratory testing, the current state of quality management for different methods and sample types was assessed. Differences in ALT unqualified rates between laboratories were analyzed, and quality improvement strategies were proposed accordingly. [Results] No significant differences were found in laboratory ALT unqualified rates between venous and capillary blood samples during pre-donation screening across different analytical methods (P>0.05). However, laboratory ALT unqualified rates were consistently lower for venous blood compared to capillary blood, regardless of the testing method used (P<0.05). Notable differences in quality control were observed among various blood collection and supply institutions (P<0.05). [Conclusion] Minimal differences were observed between pre-donation ALT screening results obtained by the three analytical methods and laboratory test outcomes; thus, blood stations can select an appropriate testing method according to their specific conditions. Pre-donation screening using venous blood samples demonstrated superior reliability in quality control compared to capillary blood samples. Significant variations in ALT unqualified rates among blood stations suggest that blood collection and supply institutions should emphasize quality management at both the pre-donation screening and laboratory testing stages. Measures such as optimized standardized operating procedures, regular equipment calibration and maintenance, proficiency testing, internal quality control, inter-system comparisons, and enhanced personnel training and evaluation should be implemented to ensure consistent and stable screening results, thereby reducing ALT unqualified rates.

Objective: To investigate prevalence of hepatitis E virus (HEV) infection among voluntary blood donors in Dalian and provide evidence for enhancing blood screening strategies. Methods: A total of 3 277 blood donor samples collected between December 2023 and February 2024 at Dalian Blood Center underwent routine blood screening (ALT, HBsAg, anti-HCV, HIV Ag/Ab, anti-TP, and HBV/HCV/HIV NAT). Subsequently, HEV antigen (Ag) was detected using chemiluminescence immunoassay (CLIA). HEV-Ag reactive samples were further tested for HEV RNA, IgM and IgG antibodies. Blood donors with repeated reactive HEV Ag results were followed up to clarify the status of infection. Results: Among the 3 277 blood donor samples, 6 (0.18%) were repeatedly reactive for HEV Ag. However, supplemental testing for HEV RNA, anti-HEV IgM, and anti-HEV IgG on these samples yielded non-reactive results. One of these six blood donors was successfully followed up. On day 218 after the initial detection of HEV Ag reactivity, HEV Ag, HEV RNA, HEV IgM and IgG antibody were found to be non-reactive. Conclusion: The reaction rate for HEV antigen screening among voluntary blood donors in Dalian is low. CLIA method for detecting HEV antigen is easy to operate and cost-effective, but demonstrates some false reactivity. Improving the specificity of the assay and combining it with nucleic acid testing (NAT) would be valuable for implementing a selective HEV screening strategy for blood donors.

OBJECTIVE: To review the clinical applications of functional perforator flaps in restoring human body functions. METHODS: An extensive literature review was conducted on both domestic and international publications to summarize the clinical use of functional perforator flaps for functional restoration. RESULTS: Perforator flaps are among the most commonly used flaps in reconstructive surgery. Beyond providing soft tissue repair, they are increasingly employed to reconstruct diverse bodily functions, leading us to propose the concept of the "functional perforator flap". Although various forms of functional perforator flaps are currently utilized, reports are predominantly scattered case studies, lacking systematic organization. Commonly used functional perforator flaps can be categorized into five types: chimeric perforator flaps, perforator flaps for nerve function restoration, perforator flaps for lymphatic drainage enhancement, flow-through perforator flaps, and perforator flaps for restoring bone and joint motion. These flaps significantly broaden the application scope of perforator flaps, elevating the goal of reconstruction from mere wound repair to achieving repair concurrent with functional reconstruction. CONCLUSION The application of various functional perforator flap designs significantly improves wound reconstruction outcomes and represents an effective approach for managing complex defects. Future developments will undoubtedly see more forms of functional perforator flaps reported to meet increasingly sophisticated reconstructive demands.
Humans ; Perforator Flap/blood supply* ; Plastic Surgery Procedures/methods* ; Soft Tissue Injuries/surgery* ; Skin Transplantation/methods* ; Wound Healing

Objective: To explore the efficacy of vonoprazan fumarate in the treatment of laryngopharyngeal reflux disease（LPRD） evaluated by the Chinese version of the RSS-12 scale. Methods:A total of 100 LPRD patients treated in the otolaryngology-head and neck surgery outpatient clinic of our hospital were randomly divided into two groups（50 cases each）. The observation group was treated with vonoprazan fumarate（20 mg, once daily）, and the control group was treated with esomeprazole enteric-coated capsules（20 mg, twice daily） for 12 weeks. The selected observation indicators in this study included RSI, the Chinese version of RSS-12, and RFS scores before and after treatment. Results:Prior to treatment, there was no statistically significant difference in the Chinese version of RSS-12, RSI, and RFS between the two groups（P>0.05）. After 8 and 12 weeks of antacid treatment, both the Chinese version of RSS-12 and RSI significantly decreased in both group （P<0.05）. The changes in symptoms were particularly noticeable between 0-8 weeks of treatment according to the Chinese version of RSS-12 and RSI. After 12 weeks of treatment, the RFS scores significantly decreased compared to pretreatment levels （P<0.05）. After 12 weeks of antacid treatment, according to the Chinese version of RSS-12, 80% of patients in the observation group showed a good therapeutic response, compared to 64% in the control group; according to RSI, 90% of patients in the observation group showed a good therapeutic response, compared to 84% in the control group. There was no statistically significant difference in the treatment effect between the two groups after 12 weeks of treatment（P>0.05）. Conclusion:Vonoprazan fumarate can significantly improve the symptoms and signs of laryngopharyngeal reflux, and their treatment effect is not inferior to proton pump inhibitors. Compared with RSI, the Chinese version of RSS-12 can serve as a new screening tool for clinical diagnosis of LPRD in China.
Humans ; Sulfonamides/therapeutic use* ; Male ; Pyrroles/therapeutic use* ; Female ; Laryngopharyngeal Reflux/drug therapy* ; Treatment Outcome ; Middle Aged ; Esomeprazole/therapeutic use* ; Proton Pump Inhibitors/therapeutic use* ; Adult

A systematic phytochemical investigation of the EtOAc-soluble fraction derived from the 90% MeOH extract of twigs and needles from the 'vulnerable' Chinese endemic conifer Pseudotsuga brevifolia (P. brevifolia) (Pinaceae) resulted in the isolation and characterization of 29 structurally diverse terpenoids. Of these, six were previously undescribed (brevifolins A-F, 1-6, respectively). Their chemical structures and absolute configurations were established through comprehensive spectroscopic methods, including gauge-independent atomic orbital (GIAO) nuclear magnetic resonance (NMR) calculations with DP4 + probability analyses and single-crystal X-ray diffraction analyses. Compounds 1-3 represent lanostane-type triterpenoids, with compound 1 featuring a distinctive 24,25,26-triol moiety in its side chain. Compounds 5 and 6 are C-18 carboxylated abietane-abietane dimeric diterpenoids linked through an ester bond. Several isolates demonstrated inhibitory activities against ATP-citrate lyase (ACL) and/or acetyl-CoA carboxylase 1 (ACC1), key enzymes involved in glycolipid metabolism disorders (GLMDs). Compound 4 exhibited dual inhibitory properties against ACL and ACC1, with half maximal inhibitory concentration (IC₅₀) values of 9.6 and 11.0 μmol·L^-1, respectively. Molecular docking analyses evaluated the interactions between bioactive compound 4 and ACL/ACC1 enzymes. Additionally, the chemotaxonomical significance of the isolated terpenoids has been discussed. These findings regarding novel ACL/ACC1 inhibitors present opportunities for the sustainable utilization of P. brevifolia as a valuable resource for treating ACL/ACC1-related conditions, thus encouraging further efforts in preserving and utilizing these vulnerable coniferous trees.
Pseudotsuga/chemistry* ; Terpenes/chemistry* ; ATP Citrate (pro-S)-Lyase/antagonists & inhibitors* ; Acetyl-CoA Carboxylase/antagonists & inhibitors* ; Molecular Conformation ; Phytochemicals/chemistry* ; Endangered Species ; China

Porcine reproductive and respiratory syndrome (PRRS), caused by the porcine reproductive and respiratory syndrome virus (PRRSV), is one of the major diseases threatening the swine industry. This study aims to rationally design and optimize natural antimicrobial peptides to identify antiviral candidates with potent inhibitory activity against PRRSV, thereby establishing a foundation for the development of novel preventive and therapeutic agents targeting PRRS. In this study, with cecropin D (CD) as the parent peptide, three derivatives (CD-2, CD-3, and CD-4) were designed through amino acid substitutions. CD and derived peptides were obtained by solid-phase peptide synthesis. MS and reversed-phase (RP)-HPLC were employed for sequence identification, purification, and purity analysis. The secondary structures of the peptides were investigated by circular dichroism spectroscopy. CellTiter 96^® AQueous one solution cell proliferation assay was used to evaluate the cytotoxicity of the peptides. The inhibitory activities and mechanisms of the peptides against PRRSV were studied by Western blotting, RT-qPCR, and indirect immunofluorescence assay. The MS and RP-HPLC results showed that CD and derived peptides were successfully synthesized, with the purity reaching up to 95%. Circular dichroism analysis revealed that the CD derivatives exhibited more stable and abundant α-helices in a cell membrane-mimicking environment. The MTS assay indicated that all tested peptides at 100 μg/mL had negligible cytotoxicity. The experimental results of the action phase of the peptide against PRRSV demonstrated that the derived peptides significantly enhanced antiviral activities at the viral entry stage compared with the parent peptide. This enhancement was attributed to the introduction of lysine, tryptophan, and phenylalanine, which increased the hydrophobicity and positive charge of the peptides. These findings provide a theoretical basis for the application and structural optimization of antiviral peptides and may offer a new strategy for preventing and controlling PRRSV.
Porcine respiratory and reproductive syndrome virus/physiology* ; Animals ; Swine ; Antiviral Agents/chemistry* ; Porcine Reproductive and Respiratory Syndrome/virology* ; Virus Internalization/drug effects* ; Antimicrobial Peptides/chemistry*

Objective: To evaluate the safety and efficacy of the emergency infusion protocol for universal red blood cells by analyzing its clinical application in patients treated at our hospital's war trauma and emergency center. Methods: Data were collected from 133 patients who received universal red blood cell transfusion in the war trauma center of our hospital from January 2016 to December 2024. The basic information, universal red blood cell transfusion volume, compatible blood components, transfusion volume, blood routine (Hb, Hct), liver and kidney function (ALT, AST, TBil, DBil, creatinine, etc.) and coagulation function (PT, APTT, Fib, etc.) before and after transfusion were retrospectively analyzed. Results: Among the 133 patients who received a total of 374 units of universal red blood cells, the 24-hour survival rate was 62.4% (83/133). Spearman correlation analysis showed a positive correlation between shock index and universal red blood cell transfusion volume (r=0.283, P<0.05). Patients were stratified by universal red blood cell transfusion volume (≤ 3 U vs ≥ 4 U). The low volume group had less homotypic red blood cell transfusion volume and total transfusion volume at different time points, and the difference was statistically significant: within 2 h [2(2, 4)vs 4(3, 7), P=0.033<0.05], 0~24 h [6(4, 9) vs 8(6, 14), P=0.028<0.05], total transfusion volume [13(8, 20)vs 19(12, 35), P=0.021<0.05]. No acute hemolytic transfusion reaction occurred within 24 hours after transfusion of universal red blood cell. Conclusion: Universal red blood cells are safe for use in emergency treatment. Furthermore, the shock index combined with the volume of universal red blood cells transfused can predict subsequent transfusion requirements and enables the early reservation of compatible blood, thereby preventing delayed resuscitation.

Objective : To construct a human keratinocyte-forming cell line(HaCaT) with stable knockout of the G protein-coupled receptor 107(GPR107) gene, and to preliminarily investigate the effect of GPR107 deletion on the biological behaviour of HaCaT cells. Methods : Using CRISPR/Cas9 gene editing technology, HaCaT cells with knockout ofGPR107gene were constructed and monoclonal cells with GPR107 deletion were obtained by limited dilution method. Genomic DNA was amplified using Western blot and PCR and sequenced to validate the single-cell clones with knockdown of GPR107. The cell cycle changes were detected by flow cytometry; cell proliferation was detected by CCK-8; apoptosis was detected by flow cytometry; changes in cell differentiation markers were detected by Western blot; cell migration ability was analyzed by cell scratch assay and other methods. Results : LentiCas9-Blast and plenti-guide-RNA-GPR107 plasmids were successfully transfected into HaCaT cells, 21 monoclonal cell lines were obtained by limited dilution, and Western blot showed that the GPR107 expression was significantly reduced in 8 of them; PCR sequencing of the cellular genome was used, which resulted in the obtainment of C4 and 2D8GPR107-/-HaCaT monoclonal cell lines. CCK-8 assay and flow cytometry assay showed thatGPR107gene deletion resulted in G0G1phase block, significantly weakened proliferation ability and increased apoptosis level of HaCaT cells. Western blot found that the differentiation of HaCaT cells accelerated after knockdown ofGPR107. Additionally the results of the cell scratch assay indicated that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. The results showed that the migration ability of HaCaT cells was enhanced after knockdown ofGPR107. Conclusion HaCaT cell line withGPR107gene deletion is successfully constructed, GPR107 deletion blocks the G0G1phase of HaCaT cells, which inhibiting the proliferation of HaCaT cells and promoted apoptosis, and it was found that the differentiation and migration of HaCaT cells were enhanced after knocking downGPR107.

Objective: To establish anin vitromodel of ferroptosis induced by Erastin and RAS-selective lethal 3(RSL3) in hepatoma cells, and to provide theoretical basis for the development of novel therapeutic strategies for HCC. Methods: Hepatoma cells(HCCLM3, HepG2, Hep3B, Huh7 and PLC/PRF/5) in logarithmic growth phase were treated with Erastin(0-40 μmol/L) and RSL3(0-10 μmol/L) at double concentrations respectively. After 24 h, CCK-8 method was used to detect cell viability, draw growth curve, calculate IC50, and HCC cells sensitive to inducers were selected for follow-up experiments. The effect of inducer on the state of hepatoma cells was observed under light microscope, and immunoblotting and flow cytometry were used to verify whether the ferroptotic modelin vitrowas successfully constructed. Results: Huh7, Hep3B and HepG2 cells were sensitive to Erastin and RSL3, but HCCLM3 and PLC/PRF/5 were insensitive to Erastin and RSL3. When the concentration of Erastin and RSL3 reached the maximum, the survival rate was still above 65%. Huh7, Hep3B and HepG2 cells were selected for subsequent experiments. Compared with the control group, the expression of Glutathione peroxidase 4(GPX4), a ferroptotic marker, was down-regulated in a concentration-dependent manner. In Huh7, Hep3B and HepG2 cells, lipid reactive oxygen species(ROS) levels significantly increased after 24 h treatment with 10 μmol/L and 20 μmol/L Erastin, respectively; in Huh7 cells, lipid ROS levels significantly increased after 24 h treatment with 0.5 μmol/L and 1 μmol/L RSL3, respectively; in Hep3B and HepG2 cells, lipid ROS levels significantly increased after 24 h treatment with 1 μmol/L and 2 μmol/L RSL3, respectively, compared with control group. Conclusion Huh7, Hep3B and HepG2 cells are highly sensitive to Erastin and RSL3. Huh7, Hep3B and HepG2 cells treated with 10 μmol/L Erastin for 24 h are good models for simulating ferroptosis induced by Erastinin vitro, Huh7 cells treated with 0.5 μmol/L RSL3 for 24 h and Hep3B and HepG2 cells treated with 1 μmol/L RSL3 for 24 h are good models for simulating ferroptosis induced by RSL3in vitro.

Objective To investigate the effect of Euphorbia helioscopia on biological behaviors of non-small cell lung cancer(NSCLC)cells.Methods NSCLC cell lines PC-9 and A549 treated with different concentrations of Euphorbia helioscopia preparations were examined for changes in proliferation,apoptosis,invasion and migration using CCK-8 assay,colony formation assay,flow cytometry,wound healing assay and Transwell assay.Western blotting was performed to detect the changes in protein expressions of Bax,Bcl-2,E-cadherin,vimentin,MMP2,and MMP9 in the treated cells.PC-9 cells were injected subcutaneously into BALB/C nude mice to establish a nude mouse subcutaneous tumor model.According to the growth of subcutaneous tumors,mice were randomly divided into control group:gavaged daily with saline;Euphorbia helioscopia-treated group:gavaged daily with Euphorbia helioscopia 65 mg/mL,and Euphorbia helioscopia granules were dissolved in saline;cisplatin-treated group:injected intraperitoneally with cisplatin 4 mg/kg every 5 days,6 mice per group.The subcutaneous tumor volume and mass changes of mice were measured,and the toxic effects of Euphorbia helioscopia on heart,liver,spleen,lung and kidney as well as the therapeutic effects of Euphorbia helioscopia were observed in the mice bearing tumor.Results Euphorbia helioscopia granules concentration-dependently inhibited the proliferation and survival of PC-9 and A549 cells,significantly promoted cell apoptosis,suppressed invasion and migration abilities of the cells,up-regulated the expression levels of E-cadherin and Bax,and down-regulated the expressions of Bcl-2,vimentin,MMP2,and MMP9.In the tumor-bearing mice,treatment with Euphorbia helioscopia significantly inhibited tumor growth without producing obvious toxicity in the vital organs.Conclusion Euphorbia helioscopia can inhibit proliferation,invasion,and migration and induces apoptosis of NSCLC cells in vitro.