Objective: To investigate the protective effects and underlying mechanisms of sodium pyruvate (SP) on RBC storage lesions using an oxidative damage model. Methods: Six units of leukocyte-depleted suspended RBCs (discarded for non-infectious reasons within three days post-collection) were randomly assigned to four groups: negative control (NS), positive control (PS), experimental group 1 (SP1), and experimental group 2 (SP2). Oxidative stress was induced in the PS group by the addition of hydrogen peroxide (H O ), while SP1 and SP2 received SP supplementation at different concentrations (25 mM and 50 mM, respectively) in the presence of H O . After 1 hour of incubation, RBC morphology was assessed microscopically, and biochemical indicators including glutathione (GSH), malondialdehyde (MDA), methemoglobin (MetHb), adenosine triphosphate (ATP), and Na /K -ATPase activity were measured. Results: RBCs in the PS group exhibited pronounced morphological damage, including cell shrinkage and echinocyte formation, whereas both SP-treated groups showed significantly reduced structural injury. SP treatment led to elevated GSH levels and decreased concentrations of MDA and MetHb, suggesting attenuation of oxidative stress. Additionally, SP enhanced intracellular ATP levels and Na /K -ATPase activity, thereby contributing to membrane stability. Notably, the SP2 group (50 mM) demonstrated superior protective effects compared to SP1 (25 mM). Conclusion: Sodium pyruvate effectively attenuates oxidative storage lesions in RBCs, primarily through its antioxidant properties, energy metabolism supporting ability, and celluar membrane stabilizing function. These findings suggest SP as a promising additive for enhancing the quality and safety of stored RBCs.

The gene GeDRP1E encoding dynamin-related protein 1E in Gastrodia elata was cloned by specific primers which were designed based on the transcriptome data of G. elata. Bioinformatics analysis on GeDRP1E gene was carried out by using ExPASy, ClustalW, MEGA, etc. Positive transgenic Arabidopsis plant and potato minituber were obtained with the genetic transformation system of Arabidopsis and potato. The plant height and seed setting rate of transgenic Arabidopsis, and agronomic characters, such as size, weight and starch content of potato minituber of transgenic potato were tested and analyzed. And GeDRP1E gene function was preliminarily investigated. The results showed that the open reading frame of GeDRP1E gene was 1 899 bp in length and 632 amino acids residues were encoded, with a relative molecular weight of 69.90 kDa and a molecular formula of C₃₀₇₉H₄₉₇₃N₈₈₃O₉₃₃S₁₉. It was predicted that the theoretical isoelectric point was 7.27, the instability coefficient was 43.34, and the average hydrophilicity index was -0.259, which was indicative of an unstable hydrophilic protein. GeDRP1E has no transmembrane structure and signal peptide, and was localized in the cytoplasm. The phylogenetic tree showed that GeDRP1E was highly homologous with DRP1E proteins of other plant species, among which GeDRP1E had the highest homology with DcDRP1E (XP_020689662.1) in Dendrobium candidum, reaching 90.05%. GeDRP1E plant expression vector pCambia1300-35Spro-GeDRP1E was constructed by double digests, and Arabidopsis complementary mutant and potato overexpression strain of GeDRP1E gene were obtained by Agrobacterium-mediated gene transformation. Compared with the Arabidopsis AtDRP1E mutant, the height and seed setting rate of the GeDRP1E complementation mutant were rescued. The minituber of GeDRP1E overexpression potato had larger size, heavier weight and higher starch content, comparing to wild-type potato. It was preliminarily induced that GeDRP1E was involved in mitochondrial morphology regulation, which related to the growth and development of Arabidopsis plants and potato miniature. The research results laid a foundation for further elucidating the molecular mechanisms underlying the growth and development of G. elata tuber development.

Liver fibrosis is a significant health burden,marked by the consistent deposition of collagen.Unfortunately,the currently available treatment approaches for this condition are far from optimal.Lysyl oxidase-like protein 2(LOXL2)secreted by hepatic stellate cells(HSCs)is a crucial player in the cross-linking of matrix collagen and is a significant target for treating liver fibrosis.Mesenchymal stem cell-derived small extracellular vesicles(MSC-sEVs)have been proposed as a potential treatment option for chronic liver disorders.Previous studies have found that MSC-sEV can be used for microRNA delivery into target cells or tissues.It is currently unclear whether microRNA-4465(miR-4465)can target LOXL2 and inhibit HSC activation.Additionally,it is uncertain whether MSC-sEV can be utilized as a gene therapy vector to carry miR-4465 and effectively inhibit the progression of liver fibrosis.This study explored the effect of miR-4465-modified MSC-sEV(MSC-sEVmiR-4465)on LOXL2 expression and liver fibrosis development.The results showed that miR-4465 can bind specifically to the promoter of the LOXL2 gene in HSC.Moreover,MSC-sEVmiR-4465 inhibited HSC activation and collagen expression by downregulating LOXL2 expression in vitro.MSC-sEVmiR-4465 injection could reduce HSC activation and collagen deposition in the CCl4-induced mouse model.MSC-sEVmiR-4465 mediating via LOXL2 also hindered the migration and invasion of HepG2 cells.In conclusion,we found that MSC-sEV can deliver miR-4465 into HSC to alleviate liver fibrosis via altering LOXL2,which might provide a promising therapeutic strategy for liver diseases.

[摘 要] 目的：探究泛素特异性蛋白酶21（USP21）在胆管癌（CCA）中的表达及其对CCA细胞增殖与迁移的作用及其机制。方法：通过生物信息学方法和免疫组化及WB法检测CCA组织及细胞中USP21的表达情况。利用体外克隆形成、EdU及Transwell实验检测敲低USP21对CCA细胞QBC939和RBE增殖及迁移的影响。通过RNA测序、质谱、免疫共沉淀（Co-IP）及WB法探究USP21的促癌机制。结果：TCGA等数据库分析结果显示，USP21 mRNA在CCA组织中呈高表达（均P < 0.05）。USP21蛋白在CCA组织和细胞中呈高表达（P < 0.05或P < 0.001或P < 0.000 1）。敲低USP21后，QBC939和RBE细胞的增殖和迁移能力均显著降低（P < 0.01或P < 0.001）。RNA测序结果表明，敲低USP21可以通过抑制PI3K/AKT信号通路抑制CCA细胞的增殖和迁移能力（P < 0.05）。质谱鉴定发现，USP21与胰岛素样生长因子2 mRNA结合蛋白1（IGF2BP1）相结合。Co-IP和WB实验结果表明，USP21与IGF2BP1结合并通过泛素化途径调控IGF2BP1的蛋白表达（P < 0.001或P < 0.000 1）。结论：USP21在CCA组织和细胞中均呈高表达，其通过IGF2BP1/PI3K/AKT信号通路增强CCA细胞的增殖及迁移能力。

Objective To investigate the benefits of using a stapler tractor in the treatment of segmental bronchus during lung segmentectomy through detailed video replay analysis of surgical procedures. Methods We collected data from patients who underwent segmentectomy performed by the same surgical team in the Department of Thoracic Surgery, the First Affiliated Hospital of Nanjing Medical University, from November 2020 to August 2023. After excluding data that lacked analysis parameters, the remaining patients were divided into four groups based on the methods used for dissociating segmental bronchus: a stapler tractor group (group A), a stapler with bronchial stretching group (group B), a stapler only group (group C), and a silk ligature group (group D). Then, we compared baseline data and videotaped surgical details across all groups. Surgical details included the success rate of one-time segmental bronchus dissociation and severance, the time taken for successful one-time dissociation and severance of the segmental bronchus, the incidence of bleeding during bronchus dissociation, the conversion rate to thoracotomy during surgery, and surgical outcomes such as total operative time, postoperative hospitalization days, postoperative thoracic drainage volume, and pulmonary air leakage rate. Results The study included 325 patients (203 in the group A, 62 in the group B, 29 in the group C, and 31 in the group D). There was no statistically significant difference in baseline data among the four groups. However, significant differences were found in terms of total operation time, postoperative hospitalization days, intraoperative blood loss, segmental bronchial stump length, postoperative air leakage rate, hemorrhage rate during segmental bronchial dissociation, and conversion to thoracotomy rate among the four groups (P<0.05). Conclusion Using a stapler tractor for dissociating segmental bronchus in lung segmentectomy results in shorter operative time, less risk of intraoperative bleeding, and less surgical complications. This study provides valuable evaluation methodologies through the analysis of video replay surgical details, contributing to the improvement of lung segmentectomy quality.

Objective To compare the consistency between the estimated values of nine common resting energy expenditure estimation equations and the measured values among the elderly population in pension institutions. Methods A total of 181 elderly individuals from Henan Guangda Ouan Leling Nursing Center were recruited as study objects through cluster sampling. The differences between the estimated values of nine common equations and the measured values obtained by indirect calorimetry were compared, and subgroup analyses were conducted to compare their consistency across different subgroups. Results Among the nine equations, the Cunningham equation and the Owen equation showed relatively small deviations from the measured values, with no significant differences (deviation rates of -1.9% and -2.0%, root mean square errors of 222.7 kcal/d and 239.4 kcal/d, and coincidence rates of 48.4% and 47.5%, respectively). In subgroups stratified by gender and body mass index (BMI), the Cunningham equation showed no significant difference compared with the measured values. The Owen equation showed no significant difference compared with the measured values only in some subgroups [males with BMI of 20~ < 27 kg/m² and ≥27 kg/m², and females with BMI < 20 kg/m²]. Conclusion The Cunningham equation, which utilizes body composition data, can assess the resting energy expenditure of the elderly in pension institutions with relatively high accuracy and speed, demonstrating strong applicability in this population.

Cough is a common symptom of several respiratory diseases. However, frequent coughing from acute to chronic often causes great pain to patients. It may turn into cough variant asthma, which seriously affects people's quality of life. For cough treatment, it is dominated by over-the-counter antitussive drugs, such as asmeton, but most currently available antitussive drugs have serious side effects. Thus, there is a great need for the development of new drugs with potent cough suppressant. BALB/c mice were used to construct mice model with cough to investigate the pharmacological effects of pectolinarigenin (PEC). Hematoxylin-eosin and Masson staining were used to assess lung injury and airway remodeling, and ELISA was used to assess the level of inflammatory factor release. In addition, inflammatory cell counts were measured to assess airway inflammation. Airway hyperresponsiveness assay was used to assess respiratory resistance in mice. Finally, we used Western blotting to explore the potential mechanisms of PEC. We found that PEC could alleviate lung tissue injury and reduce the release of inflammatory factors, inhibit of cough frequency and airway wall collagen deposition in mice model with cough. Meanwhile, PEC inhibited the Ras/ERK/c-Fos pathway to exhibit antitussive effect. Therefore, PEC may be a potential drug for cough suppression.

Objective To summarize the clinical characteristics of patients with acute posterior circulation large artery occlusive stroke within 6 hours via endovascular therapy and analyze the risk factors for poor prognosis. Methods Clinical data of 43 patients with acute posterior circulation large artery occlusive stroke within 6 hours from January 2017 to June 2023 in the Department of Neurosurgery of the Affiliated People's Hospital of Jiangsu University were collected. The baseline data, the vascular recanalization rate, symptomatic intracranial hemorrhage rate, 90 d good prognosis [modified Rankin Scale (mRS) score≤2] rate, and mortality were analyzed retrospectively. Univariate and multivariate Logistic regression analyses were applied to analyze the risk factors associated with poor prognosis (mRS score>2). Results After endovascular treatment, successful revascularization was achieved in 34 cases, and 4 cases developed symptomatic intracranial hemorrhage. At the 90-day follow-up, 20 patients had good outcomes, 12 had poor outcomes, and 11 died. Univariate analysis suggested that there was a statistically significant difference in preoperative NIHSS scores between the two groups (P < 0.05). Binary logistic regression analysis showed that a high preoperative NIHSS score was an independent risk factor for poor prognosis. Conclusion Early endovascular treatment can significantly improve the revascularization rate and prognosis of patients with acute posterior circulation large artery occlusion stroke. Preoperative NIHSS score can be used as an independent influencing factor to predict the prognosis of patients.

The gene GeDTC encoding the dicarboxylate-tricarboxylate carrier protein in Gastrodia elata was cloned by specific primers which were designed based on the transcriptome data of G. elata. Bioinformatics analysis on GeDTC gene was carried out by using ExPASY, ClustalW, MEGA, etc. Positive transgenic plants and potato minituber were obtained by virtue of the potato genetic transformation system. Agronomic characters, such as size, weight, organic acid content, and starch content, of potato minituber were tested and analyzed and GeDTC gene function was preliminarily investigated. The results showed that the open reading frame of GeDTC gene was 981 bp in length and 326 amino acid residues were encoded, with a relative molecular weight of 35.01 kDa. It was predicted that the theoretical isoelectric point of GeDTC protein was 9.83, the instability coefficient was 27.88, and the average index of hydrophilicity was 0.104, which was indicative of a stable hydrophilic protein. GeDTC protein had a transmembrane structure and no signal peptide and was located in the inner membrane of mitochondria. The phylogenetic tree showed that GeDTC was highly homologous with DTC proteins of other plant species, among which GeDTC had the highest homology with DcDTC(XP＿020675804.1) in Dendrobium candidum, reaching 85.89%. GeDTC overexpression vector pCambia1300-35Spro-GeDTC was constructed by double digests, and transgenic potato plants were obtained by Agrobacterium-mediated gene transformation. Compared with the wild-type plants, transgenic potato minituber harvested by transplanting had smaller size, lighter weight, lower organic acid content, and no significant difference in starch content. It is preliminarily induced that GeDTC is the efflux channel of tricarboxylate and related to the tuber development, which lays a foundation for further elucidating the molecular mechanism of G. elata tuber development.
Gastrodia/genetics* ; Phylogeny ; Amino Acids ; Cloning, Molecular

Humans ; M Cells ; Cell Line ; Liver Neoplasms ; Cholesterol ; Cell Line, Tumor ; Apoptosis ; Cell Cycle Checkpoints