1.Effects of resistance training on quadriceps mass and knee joint function in patients with osteoporosis and sarcopenia
Jian ZHOU ; Tao ZHANG ; Weili ZHOU ; Xingcheng ZHAO ; Jun WANG ; Jie SHEN ; Li QIAN ; Ming LU
Chinese Journal of Tissue Engineering Research 2026;30(5):1081-1088
BACKGROUND:The quadriceps strength of patients with osteoporosis and sarcopenia is significantly reduced,which can further reduce the function of the knee joint,affect the function of the lower limbs and even lead to a decrease in whole-body coordination.It is speculated that a reasonable quadriceps training program and personalized guidance are beneficial to the recovery of knee joint function in patients with osteoporosis and sarcopenia.OBJECTIVE:To observe the effect of short-term moderate-intensity resistance rehabilitation training on the mass and function of the quadriceps and knee joint function in patients with osteoporosis and sarcopenia.METHODS:Using the integrated physical examination and rehabilitation model,375 patients with osteoporosis and sarcopenia were screened at the Health Management Center of Shanghai Public Health Clinical Center.They underwent 12 weeks of combined/comprehensive exercise rehabilitation based on resistance exercise,including quadriceps resistance isotonic and isometric contraction training twice a week(3-5 sets each time,10-15 minutes per set)and aerobic exercise/balance exercise two or three times a week(30 minutes each time).Assessments and data collection were performed before rehabilitation training,12 weeks after rehabilitation training,and at follow-up 12 weeks after stopping rehabilitation training,mainly including knee joint range of motion and proprioception,quadriceps muscle strength,and cross-sectional area(magnetic resonance imaging results),pain,knee joint function(Hospital for Special Surgery score)and walking function("up-and-go"time and 6 m pace test results)as well as the patient's psychological status assessment.RESULTS AND CONCLUSION:All 375 patients completed 12 weeks of rehabilitation training and 12 weeks of follow-up without any adverse events.(1)Compared with before training,the patients' gait speed and knee range of motion increased significantly after 12 weeks of rehabilitation training(P<0.01),the time of"stand-to-walk"decreased(P<0.01),and the proprioception of the knee joint and the strength of the quadriceps femoris were significantly improved(P<0.01);and at the follow-up visit 12 weeks after stopping training,the above indicators and functions of the patients were well maintained(P>0.05).(2)Magnetic resonance imaging results showed that the effective cross-sectional area of the quadriceps femoris did not improve significantly after 12 weeks of rehabilitation training(P>0.05);but the Hospital for Special Surgery score of knee joint function increased significantly(P<0.01),and the visual analog pain scale score decreased significantly(P<0.01),suggesting that this may be related to the improvement of quadriceps femoris quality by resistance rehabilitation training.(3)The results of the Hospital Anxiety and Depression Scale score showed that the anxiety and depression scores of the patients continued to decrease,both at 12 weeks of rehabilitation training and at 12 weeks after stopping training(P<0.01).It is suggested that resistance rehabilitation training of the quadriceps can help patients with osteoporosis and sarcopenia to restore quadriceps muscle strength,increase range of motion,improve proprioception and joint stability,thereby enhancing knee joint function,reducing pain,improving depression and anxiety,and to a certain extent promoting the coordinated recovery of the musculoskeletal system.
2.miR-433 inhibits cancer stem cell-like properties and enhances 5-FU sensitivity by targeting YAP1 in colorectal cancer HCT-116 cells
DONG Xinmin1 ; ZHENG Ti2△ ; LI Tiantian3 ; JUN Mei3 ; ZHANG Jian4
Chinese Journal of Cancer Biotherapy 2026;33(5):537-544
[摘 要] 目的:探讨miR-433对结直肠癌(CRC)细胞干细胞特性及5-氟尿嘧啶(5-FU)耐药性的影响,并阐明其潜在的分子调控机制。方法: 建立5-FU耐药的HCT-116细胞株。采用RT-qPCR和WB法检测miR-433及干细胞特性标志物(SOX2、OCT4、Nanog)的表达。通过双萤光素酶报告基因实验、免疫荧光和核质分离实验验证miR-433与靶基因YAP1的关系及YAP1对其亚细胞定位的影响。通过拯救实验确认miR-433/YAP1轴的功能。利用CCK-8法检测细胞对5-FU的敏感性。结果: 与亲代HCT-116/P细胞相比,耐药的HCT-116细胞株中miR-433表达显著下调(P < 0.05),而干细胞特性标志物(SOX2、OCT4、Nanog)表达上调(P < 0.05)。过表达miR-433能够抑制干细胞标志物的表达,并直接靶向YAP1,阻碍其核定位(P < 0.05)。恢复YAP1的表达能够部分逆转miR-433对干细胞标志物的抑制作用(P < 0.05)。功能上,上调miR-433显著增强了CRC细胞对5-FU的敏感性,而此效应同样可被YAP1的重新表达所削弱(P < 0.05)。结论: miR-433通过直接靶向YAP1通路,抑制CRC细胞的干细胞特性相关分子表型,并增强其对化疗药物的敏感性。miR-433/YAP1轴可能成为克服CRC 5-FU耐药的潜在新靶点。
3.Comparative Analysis of Clinical Efficacy of Traditional Chinese Medicine Manipulative Reduction Combined with Small Splint Fixation Versus Surgical Treatment for Type A Distal Radius Fracture
Yang SHAO ; Zihan WANG ; Jianwei WANG ; Guoda DAI ; Hengyan CUI ; Zhen HUA ; Tingchen ZHU ; Shaoshuo LI ; Jun MAO ; Fenghua CHEN ; Shuai TAO ; Mao WU
Journal of Traditional Chinese Medicine 2026;67(10):1078-1085
ObjectiveTo compare the clinical efficacy of traditional Chinese medicine (TCM) manipulative reduction combined with small splint fixation versus surgical treatment for type A distal radius fracture (DRF) and to explore the factors influencing the choice of treatment. MethodsA multi-center retrospective study was conducted, collecting data from 1237 type A DRF patients treated in 11 hospitals in Jiangsu province from September, 2023 to April, 2025. Among them, 851 patients in the TCM group received manipulative reduction combined with small splint fixation, and 386 patients in the surgical group underwent open reduction and internal fixation. Visual analog scale (VAS) scores for pain and radiographic indicators including palmar tilt, ulnar deviation, and radial height were compared before treatment, 5-7 days after treatment, and 4-6 weeks after treatment. The wrist joint function scores including Dienst and Gartland-Werley scores at 12 weeks after treatment were recorded. Subgroup analysis was conducted for the excellent rate of Dienst and Gartland-Werley scores, stratified by age (<50, 50-59, 60-69, ≥70 years old) and AO subtypes (A1, A2, A3). A multivariate logistic regression model was used to identify independent factors influencing treatment choice. ResultsOn 5-7 days after treatment, the surgical group had lower VAS scores than the TCM group, while 4-6 weeks after treatment, the TCM group showed lower VAS scores than the surgical group (P<0.01). In terms of radiographic indicators, except for the palmar tilt before treatment being higher in the surgical group than in the TCM group (P<0.01), there were no significant differences in palmar tilt, ulnar deviation, and radial height at other timepoints (P>0.05). Twelve weeks after treatment, the surgical group had a higher average Gartland-Werley score and the excellent rate than the TCM group (P<0.01). Subgroup analysis showed that in patients with A2 type DRF aged 50-59 and 60-69 years old, the excellent rates of Dienst and Gartland-Werley scores in the TCM group were higher than those in the surgical group (P<0.05). Multivariate logistic regression analysis revealed that age, palmar tilt, ulnar deviation, and the degree of swelling on the affected side were independent factors influencing the choice of treatment (P<0.05). ConclusionBoth TCM manipulative reduction combined with small splint fixation and surgical treatment for type A DRF can achieve good therapeutic effects. TCM manipulative reduction combined with small splint fixation has certain advantages in medium- and long-term pain relief, especially in elderly patients, where wrist joint function recovery is more stable. Age, palmar tilt, ulnar deviation, and swelling degree are the main factors influencing the treatment choice.
4.Relationship between screen time and addictive non-suicidal self-injury behaviors among adolescent female patients with depression disorder: the mediating role of family dysfunction
Yifan LIU ; Wenle ZHANG ; Weige WU ; Jun TANG ; Yiyin HUANG ; Yun LI
Sichuan Mental Health 2026;39(2):119-125
BackgroundAddictive non-suicidal self-injury (NSSI) behaviors among adolescents have become increasingly prominent, although previous studies have identified multiple related risk factors and have examined the association between screen time and NSSI behaviors, the impact of screen time on NSSI behaviors addiction, as well as the mediating role of family dysfunction in this relationship, remain to be further clarified. ObjectiveTo investigate the mediating role of family dysfunction in the relationship between screen time and NSSI behaviors addiction among adolescent female patients with depression disorder, with the aim of providing references for reducing NSSI behaviors addiction. MethodsFrom September 2024 to November 2025, a total of 652 adolescent female patients with depression disorder were enrolled from both outpatient and inpatient departments of Xiamen Xian-yue Hospital, all of whom met the diagnostic criteria for depressive episode (F32) or recurrent depressive disorder (F33) according to the International Classification of Diseases, tenth edition (ICD-10). Assessments included a self-developed demographic questionnaire, screen use questionnaire, Chinese Family Assessment Instrument (C-FAI), and Ottawa Self-injury Inventory Chinese Revised version (OSIC). Among participants with NSSI behaviors, Spearman correlation analysis was used to examine the correlation between screen time and scale scores. Model 4 of the Process 4.1 for SPSS 26.0 was then applied to test the mediating role, and Bootstrapping procedure involving 5 000 replicates was employed to confirm the statistical significance. ResultsAmong the 652 patients, 569 (87.27%) exhibited NSSI behaviors. Among them, 398 cases (69.95%) belonged to the addictive NSSI group, and 171 cases (30.05%) belonged to the non-addictive NSSI group. The OSIC addiction dimension score was positively correlated with screen time and C-FAI scores (rs=0.114, 0.224, P<0.01). Family dysfunction mediated the relationship between screen time and NSSI addiction, with an indirect effect value of 0.036 (95% CI: 0.016–0.062), accounting for 35.88% of the total effect. ConclusionScreen time may affect the NSSI behaviors addiction in adolescent female patients with depression disorder through family dysfunction. [Funded by Joint Funds for the Innovation of Science and Technology, Fujian Province (number, 2025Y9762)]
5.The Pathogenesis and Therapeutic Strategies of Nasal Inflammatory Diseases From The Perspective of Glycolytic Metabolic Reprogramming
Meng-Wei LI ; Ji-Tang CAI ; Jun-Jie WANG ; Yi-Bo CAI ; Meng-Ting TAN
Progress in Biochemistry and Biophysics 2026;53(5):1333-1355
Aberrant activation of glycolysis represents a key metabolic mechanism underlying the initiation and progression of nasal inflammation. Allergic rhinitis, chronic rhinosinusitis, and vasomotor rhinitis exhibit distinct etiologies, yet all are characterized by inflammatory responses, impaired epithelial barrier function, and neurovascular dysregulation, in which glycolytic metabolic reprogramming acts as a central hub connecting immunometabolism and inflammatory regulation.Recent evidence indicates that glycolysis-dependent activation of immune cells provides the essential energy basis for inflammatory onset. In dendritic cells, eosinophils, mast cells, and Th2 cells, the expression of key glycolytic enzymes including HK2, PKM2, and LDHA is upregulated, thereby promoting cellular activation and proinflammatory cytokine release via the mTOR-HIF-1α signaling axis. Notably, the metabolic reprogramming of eosinophils prolongs their survival and enhances the release of cytotoxic granules, while in mast cells, enhanced glycolysis facilitates IgE-mediated degranulation and histamine release. Furthermore, glycolysis also influences the Th17/Treg balance, with enhanced glycolytic flux promoting Th17 differentiation and contributing to the heterogeneous inflammatory profiles observed across different rhinitis subtypes.As a central metabolite, lactate contributes to the formation of a metabolism-inflammation vicious cycle through multiple mechanisms. Lactate acidifies the local microenvironment to activate TRPV1 channels and facilitate neuropeptide release, mediates immune cell chemotaxis through GPR81, and regulates gene expression via histone lactylation, thereby sustaining proinflammatory gene transcription. These lactate-mediated processes collectively amplify local inflammation and contribute to the persistence of nasal symptoms.Glycolytic reprogramming in epithelial cells is modulated by the EGF/EGFR pathway, and its dysregulation may result in disrupted tight junctions, abnormal goblet cell hyperplasia, and subsequent tissue remodeling. Substance P and calcitonin gene-related peptide released from sensory neurons, in conjunction with metabolic products, synergistically maintain persistent inflammatory stimulation by activating mast cells, forming a neuro-immune-metabolic regulatory network that drives disease chronicity.From a therapeutic perspective, glycolytic inhibitors such as 2-deoxyglucose, FX11, and 3-bromopyruvate exert anti-inflammatory effects by targeting key enzymes including HK2 and LDHA, each with distinct mechanisms: 2-DG competitively inhibits hexokinase, FX11 selectively targets LDHA to reduce lactate production, and 3-BrPA modulates multiple glycolytic enzymes. Moreover, traditional Chinese medicine formulas, monomeric active components, and small-molecule compounds have shown promising potential in alleviating nasal inflammation by regulating the mTOR-HIF-1α axis, exerting antioxidant effects, and modulating endoplasmic reticulum stress pathways. The multi-target characteristics of these natural products offer advantages in addressing the complex pathophysiology of nasal inflammatory diseases.Despite these advances, several challenges remain. The non-selective inhibition of glycolysis may interfere with epithelial repair and mucosal regeneration, leading to delayed wound healing. Technical limitations in dynamic metabolic monitoring and sampling precision hinder the accurate assessment of local nasal metabolism. Furthermore, current animal models, which predominantly rely on acute stimulation protocols, inadequately recapitulate the chronic tissue remodeling processes characteristic of human rhinitis.This review systematically summarizes glycolysis as a common metabolic node shared by different rhinitis subtypes, offering a novel theoretical basis for the development of precision therapeutic strategies targeting metabolic reprogramming.
6.The Pathogenesis and Therapeutic Strategies of Nasal Inflammatory Diseases From The Perspective of Glycolytic Metabolic Reprogramming
Meng-Wei LI ; Ji-Tang CAI ; Jun-Jie WANG ; Yi-Bo CAI ; Meng-Ting TAN
Progress in Biochemistry and Biophysics 2026;53(5):1333-1355
Aberrant activation of glycolysis represents a key metabolic mechanism underlying the initiation and progression of nasal inflammation. Allergic rhinitis, chronic rhinosinusitis, and vasomotor rhinitis exhibit distinct etiologies, yet all are characterized by inflammatory responses, impaired epithelial barrier function, and neurovascular dysregulation, in which glycolytic metabolic reprogramming acts as a central hub connecting immunometabolism and inflammatory regulation.Recent evidence indicates that glycolysis-dependent activation of immune cells provides the essential energy basis for inflammatory onset. In dendritic cells, eosinophils, mast cells, and Th2 cells, the expression of key glycolytic enzymes including HK2, PKM2, and LDHA is upregulated, thereby promoting cellular activation and proinflammatory cytokine release via the mTOR-HIF-1α signaling axis. Notably, the metabolic reprogramming of eosinophils prolongs their survival and enhances the release of cytotoxic granules, while in mast cells, enhanced glycolysis facilitates IgE-mediated degranulation and histamine release. Furthermore, glycolysis also influences the Th17/Treg balance, with enhanced glycolytic flux promoting Th17 differentiation and contributing to the heterogeneous inflammatory profiles observed across different rhinitis subtypes.As a central metabolite, lactate contributes to the formation of a metabolism-inflammation vicious cycle through multiple mechanisms. Lactate acidifies the local microenvironment to activate TRPV1 channels and facilitate neuropeptide release, mediates immune cell chemotaxis through GPR81, and regulates gene expression via histone lactylation, thereby sustaining proinflammatory gene transcription. These lactate-mediated processes collectively amplify local inflammation and contribute to the persistence of nasal symptoms.Glycolytic reprogramming in epithelial cells is modulated by the EGF/EGFR pathway, and its dysregulation may result in disrupted tight junctions, abnormal goblet cell hyperplasia, and subsequent tissue remodeling. Substance P and calcitonin gene-related peptide released from sensory neurons, in conjunction with metabolic products, synergistically maintain persistent inflammatory stimulation by activating mast cells, forming a neuro-immune-metabolic regulatory network that drives disease chronicity.From a therapeutic perspective, glycolytic inhibitors such as 2-deoxyglucose, FX11, and 3-bromopyruvate exert anti-inflammatory effects by targeting key enzymes including HK2 and LDHA, each with distinct mechanisms: 2-DG competitively inhibits hexokinase, FX11 selectively targets LDHA to reduce lactate production, and 3-BrPA modulates multiple glycolytic enzymes. Moreover, traditional Chinese medicine formulas, monomeric active components, and small-molecule compounds have shown promising potential in alleviating nasal inflammation by regulating the mTOR-HIF-1α axis, exerting antioxidant effects, and modulating endoplasmic reticulum stress pathways. The multi-target characteristics of these natural products offer advantages in addressing the complex pathophysiology of nasal inflammatory diseases.Despite these advances, several challenges remain. The non-selective inhibition of glycolysis may interfere with epithelial repair and mucosal regeneration, leading to delayed wound healing. Technical limitations in dynamic metabolic monitoring and sampling precision hinder the accurate assessment of local nasal metabolism. Furthermore, current animal models, which predominantly rely on acute stimulation protocols, inadequately recapitulate the chronic tissue remodeling processes characteristic of human rhinitis.This review systematically summarizes glycolysis as a common metabolic node shared by different rhinitis subtypes, offering a novel theoretical basis for the development of precision therapeutic strategies targeting metabolic reprogramming.
7.BMP3 inhibits the inflammatory response in rat adjuvant-induced arthritis
Dantong SUN ; Xiaofeng LI ; Jun LI ; Biao SONG
Acta Universitatis Medicinalis Anhui 2026;61(3):439-447
ObjectiveTo investigate the effect of bone morphogenetic protein 3 (BMP3) on the expression of inflammatory factors and joint damage in adjuvant arthritis (AA) induced by Freund′s complete adjuvant (FCA) in rats. MethodsThe AIA model was established in SD rats by intradermal injection of FCA into the toes of the left hind limb, and BMP3 overexpressing adenovirus (Ad-BMP3) or control adenovirus (Ad-NC) was injected in situ into the knee joint cavity on day 8 after modeling. Subsequently, HE staining was used to observe the histopathological changes in the synovium, immunohistochemistry was used to detect the expression of BMP3 in the synovium, and ELISA was used to analyze the expression levels of IL-6, IL-1β and TNF-α in the serum. Primary fibroblast-like synoviocytes (FLS) were isolated from AIA rats, the expression of BMP3 in FLS was knocked down or overexpressed, and Western blot and qRT-PCR were used to detect the expression levels of BMP3 and inflammatory factors in FLS. ResultsHE staining confirmed the successful establishment of the AIA model. Compared with normal rats, AIA rats showed decreased BMP3 expression in synovial tissue. Knockdown of BMP3 promoted the protein expression of inflammatory factors (IL-6, IL-1β, IL-17A, TNF-α) and the mRNA expression of chemokines [C-C motif chemokine ligand 2 (CCL2), C-C motif chemokine ligand 3 (CCL3), Vascular Cell Adhesion Molecule-1 (VCAM-1)] in FLS. In contrast, overexpression of BMP3 suppressed the expression of these inflammatory factors and chemokines. Intra-articular injection of BMP3-overexpressing adenovirus in AIA rats upregulated BMP3 expression in synovial tissue and inhibited synovial inflammation and bone erosion. ConclusionBMP3 suppresses the production of inflammatory factors and chemokines in FLS, thereby alleviating synovial hyperplasia and bone erosion in arthritis.
8.Effect of Xingeng No.Ⅱ Granules (心梗2号颗粒剂) on Left Ventricular Remodeling in Patients of ST-Segment Elevation Myocardial Infarction with Qi Deficiency and Blood Stasis Syndrome after PCI:An Open-Labbled Randomized-Controlled Trial
Chunkun YANG ; Qinwei ZHU ; Qingquan PAN ; Jun LI
Journal of Traditional Chinese Medicine 2026;67(11):1178-1184
ObjectiveTo observe the effectiveness and safety of Xingeng No.Ⅱ Granules (心梗2号颗粒剂) in preventing and treating left ventricular remodeling in patients with qi deficiency and blood stasis syndrome following percutaneous coronary intervention (PCI) for ST-segment elevation myocardial infarction (STEMI). MethodsIn this randomized controlled trial, patients with qi deficiency and blood stasis syndrome after PCI for STEMI were randomly assigned to treatment group and control group at a 1∶1 ratio, with 66 patients in each group. In the control group, patients only received conventional western medicine after surgery, while the treatment group additionally received the granules (8 g per dose, twice daily), for a treatment duration of 8 weeks in both groups. The primary outcome was the incidence of left ventricular remodeling within 6 months after surgery. The secondary outcomes were the incidence of major adverse cardiovascular and cerebrovascular events (MACCE) within 6 months, all-cause mortality, stent thrombosis, BARC Ⅲ and V bleeding events, rehospitalization due to acute heart failure, and severe complications of STEMI. Traditional Chinese medicine (TCM) syndrome scores at 1 day and 1, 2, and 6 months after surgery was evaluated. Adverse events during the study were recorded to evaluate safety. ResultsSix cases dropped out from both the treatment group and the control group. The full analysis set (FAS) analysis showed that the incidence of left ventricular remodeling in the treatment group was 16.67% (11/66), significantly lower than 40.91% (27/66) in the control group (P=0.004). The per protocol set (PPS) analysis also showed lower incidence of left ventricular remodeling in the treatment group (20.37%, 11/54) than in the control group (49.09%, 27/55) with significant difference (P=0.002). Within 6 month after surgery, 0 patients in the treatment group and 4 out of 60 patients (6.67%) in the control group were readmitted to hospital for acute heart failure, with significantly higher rate in the control group (P=0.042). Neither group of patients experienced recurrent myocardial infarction, target vessel revascularization, in-stent thrombosis, or severe complications of STEMI. There was no statistically significant difference between groups in the incidence of stroke, cardiovascular mortality, all-cause mortality, BARC Ⅲ and V bleeding events (P>0.05). At 1 day after surgery, there was no statistically significant difference in TCM syndrome score between the groups (P>0.05); while 1, 2 and 6 months after surgery, TCM syndrome score in the treatment group was significantly lower than that in the control group (P<0.05). Analysis of the safety dataset (SS) showed that the incidence of adverse events in the treatment group was 7.41% (4/60), while in the control group it was 16.36% (9/57), showing no statistically significant difference (P = 0.117). ConclusionIn addition to conventional western medicine, Xingeng No.Ⅱ Granules can reduce the incidence of left ventricular remodeling and the incidence of rehospitalization due to acute heart failure in STEMI patients with qi deficiency and blood stasis syndrome after PCI, with good safety profile.
9.The application value of deep learning image reconstruction algorithm in ultra-low dose abdominal CT scanning
Xing TANG ; Yuncheng LI ; Hongmin SHU ; Weishu HOU ; Jun WANG ; Xiaohu LI
Acta Universitatis Medicinalis Anhui 2026;61(4):758-762
ObjectiveTo evaluate the feasibility of various strength levels of deep learning image reconstruction (DLIR) algorithms for improving non-contrast abdominal CT image quality at ultra-low radiation doses, by comparing ultra-low-dose DLIR images with low-dose filtered back projection (FBP) images. MethodsA prospective collection of 85 patients undergoing non-contrast abdominal CT scans was performed, and a self-controlled study method was employed to conduct low-dose (LD) group and ultra-low-dose (ULD) group scans. The LD group used a noise index of 10 and employed FBP for image reconstruction (LD-FBP group). The ULD group used a noise index of 30 and employed DLIR at different levels (low, medium, high), resulting in three subgroups of reconstructed images: ULD-DLIR-L, ULD-DLIR-M, and ULD-DLIR-H. For each group, CT values, standard devia-tion (SD), signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) were measured and calculated for the liver, spleen, kidneys, aorta, psoas major, and subcutaneous fat. Effective dose (ED) was also recorded. Two radiologists independently performed subjective evaluations of image quality using a 5-point scale. ResultsCompared with the LD-FBP group, the ULD-DLIR-L group showed significantly lower SNR and CNR values in the liver, spleen, kidneys, aorta, and psoas major (P<0.001), while the ULD-DLIR-H group exhibited significantly higher values (P<0.001). The difference of SNR and CNR values for the ULD-DLIR-M group showed no statistically significant difference. For subjective evaluation, the scores of the ULD-DLIR-L and ULD-DLIR-M groups were lower than those of the LD-FBP group, while there was no statistically significant difference in scores between the ULD-DLIR-H group and the LD-FBP group. The ED value of the ULD group was approximately 88% lower than that of the LD group. ConclusionCompared with the LD-FBP group, the ULD-DLIR-H group significantly reduces SD values while increasing SNR and CNR values, effectively improving the image quality of non-contrast abdominal CT scans.
10.Prokaryotic expression of Echinococcus granulosus Polo-like kinase 2 and immunoprotective efficacy of its recombinant protein
Xue WANG ; Mingzhi YAN ; Wenjing QI ; Chuanchuan WU ; Guowu ZHANG ; An GENG ; Mengxiao TIAN ; Jun LI ; Wenbao ZHANG
Chinese Journal of Schistosomiasis Control 2026;38(2):184-193
Objective To prepare the recombinant Echinococcus granulosus Polo-like kinase 2 (rEgPLK2) protein and evaluate its immunoprotective efficacy against cystic echinococcosis, so as to provide insights into research and development of novel vaccines against echinococcosis. Methods The Polo-like kinase (PLK) protein sequences were retrieved from 12 species in the NCBI protein database, including E. granulosus and E. multilocularis. Multiple sequence alignment was performed using the Clustal Omega program, and structural visualization and homology analysis were conducted using the ESPript 3.2 program. The recombinant plasmid pET-30a-EgPLK2 was transformed into BL21(DE3) competent cells. Protein expression was induced with isopropyl-β-D-thiogalactoside (IPTG), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to characterize the expression and molecular weight of the rEgPLK2 protein. The purified rEgPLK2 protein was thoroughly emulsified with Freund’s complete adjuvant at a 1 : 1 volume ratio. Two New Zealand white rabbits were immunized with multipoint subcutaneous injection on the back at a dose of 300 μg per rabbit for primary immunization. For booster immunizations, the protein was emulsified with Freund’s incomplete adjuvant at a 1 : 1 volume ratio and administered on days 14, 28, and 42 after the primary immunization at a dose of 150 μg per rabbit. Serum was sampled from the rabbit ear vein on day 7 after the final immunization to yield anti-rEgPLK2 polyclonal antibodies. Antibody titer was determined by indirect enzyme-linked immunosorbent assay (ELISA), and antibody specificity was verified by Western blotting. The tissue localization of the EgPLK2 protein was detected in E. granulosus protoscoleces and adult worms using immunofluorescence assay (IFA). Eighteen 6- to 8-week-old female SPF-grade BALB/c mice were randomly divided into three groups, including the blank control group, rEgPLK2-ISA immunization group, and PBS-ISA adjuvant control group, of 6 mice each group. Mice in the rEgPLK2-ISA immunization group and PBSISA group received three primary immunizations via intramuscular injection, and animals in the rEgPLK2-ISA immunization group was inoculated with immunogens prepared by emulsifying rEgPLK2 protein with ISA 201 adjuvant at a 1 : 1 volume ratio (6 μg per mouse), while mice in the PBS-ISA adjuvant control group received an equal volume of PBS emulsified with ISA adjuvant at a 1 : 1 volume ratio. A fourth booster immunization was administered via intraperitoneal injection. Mice in the rEgPLK2-ISA immunization group received a booster immunization with 8 μg of rEgPLK2 protein per mouse, and animals in the PBS-ISA group received an equal volume of PBS, with immunizations given at 2-week intervals. Mice in the blank control group were given no treatment, and housed under standard conditions. Tail vein blood was collected from all mice 7 days after the final immunization, and levels of specific anti-rEgPLK2 IgG antibody and its subclasses (IgG1, IgG2a, IgG2b, IgG3) were measured by indirect ELISA. E. granulosus infection was modelled in mice through injection with 1 000 E. granulosus protoscoleces via intrahepatic portal vein in the rEgPLK2-ISA immunization group and PBS-ISA adjuvant control group 2 weeks after the last immunization. All mice were sacrificed and dissected. The number of cysts was counted in mouse livers, and the cyst reduction rate was calculated. Liver tissues were processed for paraffin sectioning and stained with hematoxylin and eosin (HE), and histopathological changes were examined under a light microscope. Results Sequence analysis revealed that EgPLK2 shared a high amino acid sequence homology with E. multilocularis PLK2 (EmPLK2) and contained the typical domains of the Polo-like kinase family, including the serine/threonine protein kinase catalytic domain (STKc) and Polo-box. The IPTG-induced rEgPLK2 protein was mainly expressed in the form of inclusion bodies, and the purified rEgPLK2 protein showed a relative molecular mass of approximately 70 kDa. The prepared rabbit anti-rEgPLK2 polyclonal antibody had a titer of 1 : 256 000, and Western blotting assay showed that this anti-body specifically recognized the rEgPLK2 protein with a relative molecular mass of approximately 70 kDa. Immunofluorescence assay showed that the EgPLK2 protein was localized in the excretory bladder and rostellum of E. granulosus protoscoleces, as well as the tegument, suckers, and inter-proglottid junctions of adult worms. Immunoprotective assay showed that the serum levels of specific anti-rEgPLK2 IgG, IgG1, IgG2a, and IgG2b antibodies were 2.92 ± 0.49, 0.33 ± 0.10, 0.31 (0.36), and 3.12 (1.73) in mice in the rEgPLK2-ISA immunization group, which were all significantly higher than those in the PBS-ISA adjuvant control group (0.14 ± 0.04, 0.07 ± 0.01, 0.12 ± 0.04, and 0.11 ± 0.04, respectively) (t = 19.28 and 8.46, Z = 3.75 and 4.15; all P values < 0.001); however, there was no significant difference in the serum anti-IgG3 antibody level between the rEgPLK2-ISA immunization group and the PBS-ISA adjuvant control group [0.07 (0.01) vs. 0.073 (0.07); Z = 0.69, P > 0.05)]. In the mouse model of E. granulosus infections, the area of hepatic lesions was reduced and the inflammatory infiltration was alleviated in the rEgPLK2-ISA immunization group than in the PBS-ISA adjuvant control group, and the number of hepatic cysts was higher in the PBS-ISA adjuvant control group than in the rEgPLK2-ISA immunization group [8.00 (2.00) vs. 1.00 (0.75); Z = −2.93, P < 0.01], with a cyst reduction rate of 80.40%. Indirect ELISA assay measured higher serum levels of specific anti-rEgPLK2 IgG (3.28 ± 0.48 vs. 0.11 ± 0.04; t = 15.86, P < 0.01), IgG1 (0.29 ± 0.02 vs. 0.09 ± 0.01; t = 15.67, P < 0.01), IgG2a [3.71 (1.09) vs. 0.08 (0.03); Z = 2.88, P < 0.01], and IgG2b antibodies [3.34 (1.01) vs. 0.08 (0.03); Z = 2.88, P < 0.01] in the rEgPLK2-ISA immunization group than in the PBS-ISA adjuvant control group, and there was no significant difference in the serum level of the specific anti-rEgPLK2 IgG3 antibody between the rEgPLK2-ISA immunization group and the PBS-ISA adjuvant control group (0.07 ± 0.01 vs. 0.07 ± 0.01; t = 1.29, P > 0.05). Conclusions The prokaryotic expression system has been successfully constructed for the EgPLK2 gene and the anti-rEgPLK2 polyclonal antibody has been obtained. The rEgPLK2 protein exhibits a high immunogenicity, and is effective to protect against E. granulosus infection, and inhibits cyst development, which is a promising candidate vaccine target against cystic echinococcosis.

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