OBJECTIVE To explore the potential mechanism by which drug-containing serum of Dianxianqing granules （DXQ） inhibits microglial ferroptosis. METHODS Male SD rats were given normal saline and Dianxianqing granules solution via intragastric administration to prepare normal serum and DXQ， respectively. Mice microglia BV2 cells were collected and successfully transfected with a negative control small interfering RNA （si-NC）， and then they were included in the si-NC group and cultured under normal conditions. Cells successfully transfected with small interfering RNA targeting glutathione peroxidase 4 （GPX4） （si-GPX4） were divided into the si-GPX4 group， the CsA group （treated with 1 μmol/L cyclosporine A）， and the DXQ- L， DXQ-M and DXQ-H groups （treated with 5%， 7% and 10% DXQ， respectively）. These groups were subsequently treated with their corresponding drug solutions and ferroptosis inducer Erastin （10 μmol/L）. The intracellular levels of total iron ions， glutathione （GSH）， reactive oxygen species （ROS）， and the expression of mitochondrial superoxide were determined in each group after 48 h of treatment. Additionally， mitochondrial membrane potential， the opening degree of mitochondrial permeability transition pore （MPTP）， and mRNA expressions of GPX4 and cyclophilin D （CypD） were detected. Furthermore， the expressions of ferroptosis-related proteins[GPX4， transferrin receptor 1 （TfR1） and ferritin heavy chain 1 （FTH1）]， as well as MPTP-related proteins [adenine nucleotide translocator （ANT）， cytochrome C （CytC）， mitochondrial calcium uniporter （MCU） and CypD] were assessed. RESULTS Compared with si-NC group， the levels of total iron ions and ROS， the expression level of mitochondrial superoxide， the opening degree of MPTP， protein and its mRNA expressions of CypD as well as protein expressions of TfR1 and MCU were increased or up-regulated significantly （P＜0.01）； however， GSH content， mitochondrial membrane potential， protein and mRNA expressions of GPX4， and protein expressions of FTH1， ANT and CytC were decreased or down-regulated significantly （P＜0.01）. Compared with the si-GPX4 group， the cells in the DXQ-M， DXQ-H groups showed a general improvement in the above quantitative indicators （P＜0.01 or P＜0.05）. CONCLUSIONS DXQ can enhance antioxidant capacity by activating the GSH/GPX4 pathway， regulate the expressions of TfR1 and FTH1 protein to correct iron ion homeostasis， inhibit excessive opening of MPTP to improve mitochondrial function， and ultimately suppress microglial ferroptosis.

This review aimed to provide a comprehensive analysis of the etiology, epidemiology, pathology, and conventional treatment of heterotopic ossification (HO), especially emerging potential therapies. HO is the process of ectopic bone formation at non-skeletal sites. HO can be subdivided into two major forms, acquired and hereditary, with acquired HO predominating. Hereditary HO is a rare and life-threatening genetic disorder, but both acquired and hereditary form can cause severe complications, such as peripheral nerve entrapment, pressure ulcers, and disability if joint ankylosis develops, which heavily contributes to a reduced quality of life. Modalities have been proposed to treat HO, but none have emerged as the gold standard. Surgical excision remains the only effective modality; however, the optimal timing is controversial and may cause HO recurrence. Recently, potential therapeutic strategies have emerged that focus on the signaling pathways involved in HO, and small molecule inhibitors have been shown to be promising. Moreover, additional specific targets, such as small interfering RNAs (siRNAs) and non-coding RNAs, could be used to effectively block HO or develop combinatorial therapies for HO.
Humans ; Ossification, Heterotopic/genetics*

BACKGROUND:The occurrence of neuronal axonal injury can result in neurological dysfunction,and the facilitation of axonal elongation is anticipated to play a pivotal role in the treatment of diseases affecting the nervous system.OBJECTIVE:To investigate whether ectomesenchymal stem cells-derived extracellular vesicles can promote neuronal axonal elongation.METHODS:(1)Ectomesenchymal stem cells were obtained from nasal mucosa using the tissue adherence method,and the specific markers of were identified through immunofluorescence.Ectomesenchymal stem cells-derived extracellular vesicles were acquired via ultracentrifugation and identified.(2)Ectomesenchymal stem cells-derived extracellular vesicles(0,0.5,1.0,1.5 mg/mL)were incubated with PC12 cells for 72 hours.The cytotoxicity and proliferation of ectomesenchymal stem cells-derived extracellular vesicles on PC12 cells were assessed using the CCK-8 assay.(3)Ectomesenchymal stem cells-derived extracellular vesicles(1.0 mg/mL)were incubated with PC12 cells or neurons for 72 hours.The changes in axon length were observed using microscopic analysis.The expression levels of axon-related markers β3-tubulin(early stage),growth associated protein 43(middle stage),and neurofilament 200(mature stage)were analyzed through real-time fluorescence quantitative PCR and Western blotting.These investigations aimed to explore the potential of ectomesenchymal stem cells-derived extracellular vesicles in promoting neurite elongation within PC12 cells or neurons.RESULTS AND CONCLUSION:(1)The majority of the acquired ectomesenchymal stem cells exhibited a spindle-shaped morphology,while a minority displayed irregular shapes,and demonstrated high expression levels of mesenchymal stem cell-specific markers Nestin,CD44,and Vimentin.The obtained ectomesenchymal stem cells-derived extracellular vesicles fulfilled the biological criteria for extracellular vesicles.(2)Within the detected protein concentration range of 0.5 to 1.5 mg/mL,the proliferation of PC12 cells was promoted by ectomesenchymal stem cells-derived extracellular vesicles,and this effect was further enhanced with increasing concentrations.(3)Ectomesenchymal stem cells-derived extracellular vesicles increased the length of axons in PC12 cells and neurons and the expression of axon-related markers β3-tubulin,growth associated protein 43,and neurofilament 200.Above findings suggest that ectomesenchymal stem cells-derived extracellular vesicles have the potential to enhance neuronal axonal elongation.

Objective:To isolate Yersinia enterocolitica phage and study its biological and genomic characteristics, and explore its application value. Methods:Phages were isolated from cecum of Apodemus chevrieri in the plague focus of wild rodent in Lijiang employing Yersinia enterocolitica (CMCC52301) as the host strain. After purification, proliferation and concentration, the morphology was observed by transmission electron microscope, and the host spectrum of the phage was determined by double-layer plate method. Phage DNA was extracted and sequenced, and the raw sequences data were assembled and visually analyzed, also performed genome functional annotation, phage classification status, phylogenetic tree, genome collinearity analysis, etc. Results:One strain of Yersinia enterocolitica phage was isolated and named vB_Yen_YN301-151, which had a head and a contractile tail. The phage only lysed Yersinia enterocolitica, and could not lyse other tested strains. And at a lysis temperature of 37 ℃, more transparent plaques were observed compared to at a lysis temperature of 21 ℃. The gene structure of this phage was double-stranded DNA, with a genome length of 51 176 bp and a GC content of 46.96%, and 95 genes were predicted to encode 15 structural morphological related proteins, 9 proteins involved in replication and metabolism, 3 DNA assembly functional proteins, 1 phage lysis related protein, and 67 hypothetical proteins. It was identified that the phage belong to the Drexlerviridae family of the Caudoviricetes class, clustered with vB_YenM_534, vB_Yen_X1, vB_YenM_281, and vB_YenM_531 in the phylogenetic tree and exhibited good collinearity with PY100, vB_YenM_ 25, and vB_Yen_X1. Conclusions:The successfully isolated vB_Yen_YN301-151 is a novel phage belonging to the Drexlerviridae family, and its gene encoded proteins are mostly hypothetical proteins. It only lyses Yersinia enterocolitica, with host specificity and has the potential to be used as diagnostic phage for screening Yersinia enterocolitica.

Objective:To isolate Yersinia enterocolitica phage and study its biological and genomic characteristics, and explore its application value. Methods:Phages were isolated from cecum of Apodemus chevrieri in the plague focus of wild rodent in Lijiang employing Yersinia enterocolitica (CMCC52301) as the host strain. After purification, proliferation and concentration, the morphology was observed by transmission electron microscope, and the host spectrum of the phage was determined by double-layer plate method. Phage DNA was extracted and sequenced, and the raw sequences data were assembled and visually analyzed, also performed genome functional annotation, phage classification status, phylogenetic tree, genome collinearity analysis, etc. Results:One strain of Yersinia enterocolitica phage was isolated and named vB_Yen_YN301-151, which had a head and a contractile tail. The phage only lysed Yersinia enterocolitica, and could not lyse other tested strains. And at a lysis temperature of 37 ℃, more transparent plaques were observed compared to at a lysis temperature of 21 ℃. The gene structure of this phage was double-stranded DNA, with a genome length of 51 176 bp and a GC content of 46.96%, and 95 genes were predicted to encode 15 structural morphological related proteins, 9 proteins involved in replication and metabolism, 3 DNA assembly functional proteins, 1 phage lysis related protein, and 67 hypothetical proteins. It was identified that the phage belong to the Drexlerviridae family of the Caudoviricetes class, clustered with vB_YenM_534, vB_Yen_X1, vB_YenM_281, and vB_YenM_531 in the phylogenetic tree and exhibited good collinearity with PY100, vB_YenM_ 25, and vB_Yen_X1. Conclusions:The successfully isolated vB_Yen_YN301-151 is a novel phage belonging to the Drexlerviridae family, and its gene encoded proteins are mostly hypothetical proteins. It only lyses Yersinia enterocolitica, with host specificity and has the potential to be used as diagnostic phage for screening Yersinia enterocolitica.

BACKGROUND:The occurrence of neuronal axonal injury can result in neurological dysfunction,and the facilitation of axonal elongation is anticipated to play a pivotal role in the treatment of diseases affecting the nervous system.OBJECTIVE:To investigate whether ectomesenchymal stem cells-derived extracellular vesicles can promote neuronal axonal elongation.METHODS:(1)Ectomesenchymal stem cells were obtained from nasal mucosa using the tissue adherence method,and the specific markers of were identified through immunofluorescence.Ectomesenchymal stem cells-derived extracellular vesicles were acquired via ultracentrifugation and identified.(2)Ectomesenchymal stem cells-derived extracellular vesicles(0,0.5,1.0,1.5 mg/mL)were incubated with PC12 cells for 72 hours.The cytotoxicity and proliferation of ectomesenchymal stem cells-derived extracellular vesicles on PC12 cells were assessed using the CCK-8 assay.(3)Ectomesenchymal stem cells-derived extracellular vesicles(1.0 mg/mL)were incubated with PC12 cells or neurons for 72 hours.The changes in axon length were observed using microscopic analysis.The expression levels of axon-related markers β3-tubulin(early stage),growth associated protein 43(middle stage),and neurofilament 200(mature stage)were analyzed through real-time fluorescence quantitative PCR and Western blotting.These investigations aimed to explore the potential of ectomesenchymal stem cells-derived extracellular vesicles in promoting neurite elongation within PC12 cells or neurons.RESULTS AND CONCLUSION:(1)The majority of the acquired ectomesenchymal stem cells exhibited a spindle-shaped morphology,while a minority displayed irregular shapes,and demonstrated high expression levels of mesenchymal stem cell-specific markers Nestin,CD44,and Vimentin.The obtained ectomesenchymal stem cells-derived extracellular vesicles fulfilled the biological criteria for extracellular vesicles.(2)Within the detected protein concentration range of 0.5 to 1.5 mg/mL,the proliferation of PC12 cells was promoted by ectomesenchymal stem cells-derived extracellular vesicles,and this effect was further enhanced with increasing concentrations.(3)Ectomesenchymal stem cells-derived extracellular vesicles increased the length of axons in PC12 cells and neurons and the expression of axon-related markers β3-tubulin,growth associated protein 43,and neurofilament 200.Above findings suggest that ectomesenchymal stem cells-derived extracellular vesicles have the potential to enhance neuronal axonal elongation.

Objective:To explore the feasibility and consistency of a new digital classification system of pelvic fractures named as JST classification based on close reduction techniques.Methods:A retrospective collection was conducted of the data from the 63 patients with pelvic fracture who had undergone surgical treatment after JST classification at Department of Orthopaedics and Traumatology, Beijing Jishuitan Hospital from March 2021 to March 2023. Digital classification of the pelvic fractures was performed based on their locations and displacements. The classification first divides the pelvis into 4 parts: left half pelvis and right half pelvis; sacral Denis Ⅲ area and pubic symphysis. The symmetrical left and right sacral Denis Ⅰ and Denis Ⅱ areas are also included in the left/right half pelvis. Subsequently, the left half pelvis and right half pelvis are divided into 4 regions and marked by capitalized English letters: Sacrum Area (including Denis Ⅰ and Denis Ⅱ, denoted as S), Sacroiliac Joint Area (denoted as J), Iliac Area (denoted as I), and Pubic Area (denoted as P); to distinguish right/left, R and L are used as prefixes. The 2 asymmetric parts are also marked with English letters: Denis Ⅲ area of the sacrum (denoted as Sac), and pubic symphysis (denoted as C). Afterwards, the fracture line morphology and displacement in each region are marked digitally to form a complete JST classification system. The inter- and intra-observer reliabilities (Fleiss' and Cohen's Kappa) of the JST classification system were tested by 3 observers with more than 10 years of experience in pelvic fracture treatment.Results:Consistency analysis of the JST classification results showed that the mean κ value of the intra-observer reliability was 0.818 (from 0.658 to 0.946, P<0.001) and the inter-observer reliability 0.873 (from 0.674 to 1.000, P<0.001), both indicating excellent agreement. Of the 63 patients, 59 obtained successful closed reduction with the assistance of the Rossum Robot R-Universal intelligent orthopedic surgical robot system after fracture classification by the JST system, yielding a success rate of 93.7% (59/63). Conclusions:The new JST classification system for pelvic fractures demonstrates strong intra and inter-observer reliabilities compared with traditional classification systems. As JST classification system labels each fracture site and key bones, it is of great significance for the deep learning and intraoperative operations of intelligent fracture robots.

Objective:To study the effect of iodine excess on antioxidant capacity and blood lipid in adult.Methods:A survey was conducted in areas with different iodine nutrition levels in Shandong and Shanxi provinces to collect fasting morning urine and venous blood samples of adults. Urinary iodine, serum levels of superoxide dismutase(SOD), malondialdehyde (MDA), total cholesterol (TCHO), triglyceride (TG), apolipoprotein A1 (apoA1) and apolipoprotein B (apoB), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were determined. According to the median urinary iodine of the population in the investigated village, they were divided into appropriate iodine group (100-299 μg/L) and iodine excess group (≥300 μg/L) . Multiple linear regression was used to analyze the effects of iodine nutrition and other factors on oxidative stress indexes and blood lipids. Partial correlation analysis was used to analyze the relationship between iodine nutrition and oxidative stress indexes and blood lipids.Results:A total of 1 049 subjects were included, including 471 in the appropriate iodine group and 578 in the iodine excess group. The median (quartile) urinary iodine of the appropriate iodine group and the iodine excess group was 228.70 (157.02, 341.49) and 558.73 (298.06, 985.06) μg/L, respectively. The serum SOD level of the appropriate iodine group and the iodine excess group was 12.60 (10.83, 14.10) and 11.29 (9.18, 13.10) U/ml, respectively, and the difference between the groups was statistically significant ( U = 92 697.50, P < 0.001). There were statistically significant differences in serum TG, HDL-C and apoB levels between the appropriate iodine group and the iodine excess group ( U = 108 879.50, 96 613.50, 99 050.50, P < 0.05). The results of multiple linear regression showed that after excluding age, gender and body mass index (BMI), there was a negative correlation between iodine nutrition and serum SOD and HDL-C levels [standard regression coefficient ( β) = - 0.196, - 0.294, P < 0.001]. Partial correlation analysis showed that there was a negative correlation between iodine nutrition and serum SOD and HDL-C levels [correlation coefficient ( r) = - 0.16, - 0.09, P < 0.05]. Conclusion:Excessive iodine intake affects oxidative stress and lipid metabolism in human body.

Objective:To explore the short-term efficacy of fixation with a 3D printed individualized custom-made plate in the treatment of elderly patients with periprosthetic femoral fracture.Methods:Retrospectively analyzed were the 5 elderly patients with periprosthetic femoral fracture who had been treated by fixation with a 3D printed individualized custom-made plate from January 2022 to July 2022 at Department of Orthopaedics and Traumatology, Beijing Jishuitan Hospital. There were 3 males and 2 females, aged 81, 86, 77, 91 and 87 years, respectively. One left and 4 right limbs were affected. Vancouver classification: type B1 ( n=3), type B2 ( n=1), and type C ( n=1). The time from operation to injury was 5, 6, 10, 5 and 7 days, respectively. Preoperatively, the femur affected, prosthesis and individualized plate with a greater trochanteric hook, loop cable channel and bone-like trabecular microporous structure were custom-made by 3D printing according to 1:1 models. Virtual operations were simulated to formulate surgical protocols. The operation time, length of surgical incision, intraoperative blood loss and transfusion, hospital stay, hip function and complications at the last follow-up were recorded. Results:The 5 patients were followed up for 12, 7, 10, 3 and 6 months, respectively. There were no events of superficial incision or deep prosthesis infection. Respectively, the operation time was 1.8, 1.7, 2.3, 2.0 and 3.3 h; the length of surgical incision 31, 30, 38, 27 and 30 cm; the intraoperative bleeding volume 400, 300, 300, 500 and 600 mL; the length of hospital stay 8, 9, 15, 14 and 11 d. Four patients received intraoperative blood transfusion of 300, 900, 150 and 1, 050 mL, respectively. One patient died of a heart attack 3 months after discharge; another patient developed dyskinesia at the contralateral limb 3 months after discharge due to cerebral infarction and died of recurrent cerebral infarction 7 months after discharge. At the last follow-up, the Harris hip scores of 3 patients were 86, 77 and 69 points, respectively. None of the patients had complications like breakage or loosening of implants.Conclusion:In the treatment of elderly patients with periprosthetic femoral fracture, fixation with a 3D printed individualized custom-made plate may lead to fine limb function and good short-term curative efficacy.

Objective:To investigate the early clinical effectiveness of using customized 3D printed acetabular augment to repair large acetabular bone defects in delayed total hip arthroplasty after failed treatment of acetabular fractures.Methods:A retrospective study was conducted to analyze the 6 patients who had undergone 3D printed acetabular augment to reconstruct acetabular bone defects in delayed total hip replacement from July 2021 to January 2023 after failed treatment of acetabular fractures. They were all males, with an age of (51.3±15.0) years. Paprosky classification: 2 cases of type ⅡB, 1 case of type ⅡC, and 3 cases of type ⅢA. Recorded were surgical time, intraoperative bleeding, hospitalization time, and visual analogue scale (VAS), and modified Merle d'Aubigné & Postel score, Harris hip score, and leg length discrepancy at the last follow-up.Results:For the 6 patients, the mean surgical time was (222.5±46.9) min, the mean intraoperative bleeding 1,100 (1,000, 2,625) mL, the mean hospitalization time (9.0±4.5) d, and the mean follow-up time (11.8±7.9) months. At the last follow-up, the VAS [(2.5±1.0) points] significantly decreased compared with the preoperative value [(6.2±0.8) points], the modified Merle d'Aubigné & Postel score [(13.2±2.1) points] and Harris hip score [(67.8±15.3) points] significantly increased compared with the preoperative values [(6.7±0.8) and (34.2±8.4) points], the vertical position of center of rotation [(22.5±5.2) mm] and the horizontal position of center of rotation [(40.7±2.6) mm] were significantly reduced compared with the preoperative values [(38.1±14.2) and (53.1±10.0) mm] ( P<0.05). At the last follow-up, the leg length discrepancy was (6.2±3.6) mm, showing no statistically significant difference from the preoperative value [(18.7±1.7) mm] ( P>0.05). At the last follow-up, no clear line at the cup-bone or augment-bone interface, or no possible prosthetic loosening or displacement was observed on the X-ray films. Conclusion:In delayed total hip arthroplasty after failed treatment of acetabular fractures, the customized 3D printed augment for repair of large bone defects in the acetabulum can reconstruct the normal rotation center of the hip joint, provide reliable stability for the cup prosthesis, and enable patients to obtain significant improvements in hip function.