BACKGROUND:In endodontics,revascularization and effective control of bacterial infection are prerequisite for regenerative repair of tissues and further development of the root apex.Ornidazole,carried in pulp-capping materials or vascularized scaffolding materials may control pulpal infections,but its effect on vascularization need to be investigated.OBJECTIVE:To investigate the residual concentration pattern of ornidazole in root canals and to evaluate the effects of ornidazole on endothelial cell proliferation,migration,and differentiation,as well as on vascular irritation.METHODS:(1)Ornidazole was encapsulated in the isolated pulp cavity and then immersed in Hank's balanced salt solution for 7 days.Ornidazole was then removed from the pulp cavity,reencapsulated in sterile water,and again immersed in Hank's balanced salt solution.The mass concentration of ornidazole in the pulp cavity fluid was measured periodically by colorimetric method.(2)Human umbilical vein endothelial cells were inoculated into well plates.Adherent cells were stimulated by the addition of lipopolysaccharide for 24 hours,and then co-cultured by the addition of 0,1,2,5,8,10 μg/mL ornidazole,to detect the cellular activity and migratory ability.Human umbilical vein endothelial cells were inoculated in well plates and co-cultured with different mass concentrations(0,1,2,5,8,10 μg/mL)of ornidazole or stimulated by lipopolysaccharide for 24 hours followed by the addition of different mass concentrations(0,1,2,5,8,10 μg/mL)of ornidazole.The gene expression of vascular endothelial growth factor and basic fibroblast growth factor as well as the protein expression of vascular endothelial growth factor was detected.(3)The chorioallantoic membrane assay was employed to assess the vascular irritation of 2 and 10 μg/mL ornidazole.RESULTS AND CONCLUSION:Residual ornidazole in exfoliated teeth was rapidly released within the initial 6 days,with a subsequent decrease in release rate,maintaining a concentration of approximately 2 μg/mL at the root apex after 8 days.Under lipopolysaccharide-induced inflammatory conditions,cell counting kit-8 and cell live-dead fluorescence staining showed that ornidazole(1-10 μg/mL)had no significant effect on the activity of human umbilical vein endothelial cells,and the cell scratch assay showed that ornidazole(1-10 μg/mL)had no obvious effect on the migratory ability of human umbilical vein endothelial cells.RT-qPCR assay showed that,after co-cultivation with ornidazole alone,the mRNA expression of vascular endothelial growth factor and basic fibroblast growth factor in human umbilical vein endothelial cells showed an overall decreasing trend.After co-culturing with ornidazole under lipopolysaccharide-induced inflammation,the mRNA expression of the two factors showed a rising trend in human umbilical vein endothelial cells.Western blot assay showed that vascular endothelial growth factor protein expression had an elevating trend in human umbilical vein endothelial cells after co-culture with ornidazole under lipopolysaccharide-induced inflammatory conditions.The chorioallantoic membrane assay showed that 2 and 10 μg/mL ornidazole were non-vascular irritating.To conclude,1-10 μg/mL ornidazole is non-cytotoxic and non-vascular irritating,promotes the expression of angiogenesis-related genes and proteins in inflammatory endothelial cells,and serves as a potential therapeutic agent for pulpal infection control.

Vertebral refracture following percutaneous vertebral augmentation (PVA) is commonly seen in elderly patients with osteoporotic thoracolumbar compression fractures (OTLCF). It can lead to recurrent pain, loss of vertebral height, progression of kyphosis, and even neurological dysfunction, significantly impairing patients′ quality of life. Current diagnosis and treatment face multiple challenges, including high misdiagnosis rate, difficulty in choosing between surgical and non-surgical treatment options, lack of standardized surgical protocols, interference from intralesional bone cement during procedures, inadequate stability of internal fixation in osteoporotic bone, and suboptimal compliance of anti-osteoporotic therapy. Establishing a standardized diagnostic and therapeutic framework is urgently needed. To standardize the management process and improve outcomes for vertebral refractures after PVA in elderly OTLCF patients, Spinal Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field to develop Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures ( version 2025), based on current literature and clinical experience, and adhering to principles of scientific rigor and clinical applicability. A total of 11 recommendations were proposed, encompassing diagnosis, treatment, and rehabilitation of vertebral refracture after PVA in elderly patients with OTLCF, aiming to provide a foundation for a standardized management.

Thoracolumbar spine fracture often leads to severe pain, functional impairments, and neurological deficits, for which open reduction and internal fixation can effectively restore the spinal structural stability. Open decompression and reduction with internal fixation can help relieve spinal cord compression and improve spinal function in cases of concomitant cord injury. Although spinal stability can be restored through surgery, patients often face chronic pain and functional impairments postoperatively. A postoperative rehabilitation program is critical in optimizing therapeutic outcomes, reducing complications, and minimizing the risk of secondary injuries. However, current rehabilitation methods, such as physical therapy, functional training, and pain management, are confronted with problems in clinical practice, including significant variation in efficacy, poor patient adherence, and prolonged rehabilitation period. There is an urgent need for a unified rehabilitation strategy to address these problems. To this end, the Spinal Trauma Group of the Orthopedic Physicians Branch of the Chinese Medical Association and the Spine Health Professional Committee of the Chinese Human Health Technology Promotion Association organized experts from relevant fields to formulate Evidence-based guidelines for rehabilitation treatment after internal fixation of thoracolumbar spine fracture in adults ( version 2025) by integrating evidences from clinical researches and advanced rehabilitation concepts at home and abroad. A total number of 14 recommendations concerning the rehabilitation treatment with multimodal analgesia, psychological intervention, deep vein thrombosis prevention, core muscle and extremity exercise, appropriate use of braces, early weight-bearing, device-aided rehabilitation exercise, neuroregulatory therapy, rehabilitation team were put forward, aiming to standardize the post-operative rehabilitation process following internal fixation, promote the functional recovery, and enhance patients′ quality of life.

Acute symptomatic osteoporotic thoracolumbar compression fracture (ASOTLF) can lead to chronic low back pain, kyphosis deformity, pulmonary dysfunction, loss of mobility, and even life-threatening complications. Vertebral augmentation is currently the mainstream treatment method for this condition. In 2019, the Editorial Board of Chinese Journal of Trauma and the Spinal Trauma Group of Orthopedic Surgeons Branch of Chinese Medical Doctor Association collaboratively led the development of Clinical guideline for vertebral augmentation for acute symptomatic osteoporotic thoracolumbar compression fractures. Six years later, with advances in clinical diagnosis and treatment techniques as well as accumulating evidence in related fields, the 2019 guideline requires updating. To this end, the Spinal Trauma Group of Orthopedic Surgeons Branch of Chinese Medical Doctor Association, the Spinal Health Professional Committee of China Human Health Science and Technology Promotion Association, and the Minimally Invasive Orthopedics Professional Committee of Shaanxi Medical Doctor Association have organized experts in the field to develop the Clinical guideline for vertebral augmentation of acute symptomatic osteoporotic thoracolumbar compression fractures ( version 2025) , based on the latest evidence-based medical researches. This guideline incorporates 3 recommendations retained from the 2019 version with updated strength of evidence, along with 12 new recommendations. It provides recommendations from six aspects of diagnosis, pain management, treatment option selection, prevention of postoperative complications, anti-osteoporosis therapy, and postoperative rehabilitation, aiming to provide a reference for standard treatment of vertebral augmentation for ASOTLF in hospitals at all levels.

Objective:To assess the impact of vanishing twin syndrome (VTS) on the accuracy of non-invasive prenatal testing (NIPT).Methods:Three pregnant women who underwent NIPT testing at Guangdong Women and Children′s from November 2019 to February 2020 were selected as the study subjects. The three women had either vanish twin syndrome or had undergone fetal reduction for other reasons in one of their twins, and were subsequently subject to NIPT, chromosome karyotyping, chromosome microarray analysis (CMA), and short tandem repeat (STR) analysis. This study has been approved by the Medical Ethics Committee of Guangdong Maternal and Child Health Hospital (Ethic No.: 20230132).Results:Case 1 underwent selective fetal reduction at 8 + weeks of gestation. At 17 + weeks, NIPT showed a fetal DNA fraction of 2.806%, with results indicating the presence of Y chromosome and abnormal sex chromosome ratios. However, the women had subsequent uncomplicated vaginal delivery of a female infant, and no abnormality noted. Case 2 experienced spontaneous demise of one twin at 13 weeks′ gestation. At 19 weeks, NIPT indicated a high risk for chromosome 21 ( Z-score 4.671) in the surviving fetus, but subsequent evaluation showed no abnormality. Case 3, a dichorionic diamniotic (DCDA) twin pregnancy, underwent selective reduction at 13 + weeks due to fetal abnormalities in one twin. At 22 + weeks, NIPT for the surviving fetus indicated a high risk for chromosome 21 (Z-score 17.549), but subsequent evaluation was unremarkable. Conclusion:In twin pregnancies, the relatively low cell-free fetal DNA (cffDNA) concentration can compromise the success rate and accuracy of NIPT compared to singleton pregnancies. Residual DNA from the demised fetus may persist for weeks following VTS or selective reduction, potentially causing false-positive NIPT results and interfering with sex chromosome prediction for the surviving fetus. Additionally, determining chorionicity is critical for reliable interpretation of NIPT results in twin pregnancies.

BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.

OBJECTIVE: To assess the impact of vanishing twin syndrome (VTS) on the accuracy of non-invasive prenatal testing (NIPT). METHODS: Three pregnant women who underwent NIPT testing at Guangdong Women and Children's from November 2019 to February 2020 were selected as the study subjects. The three women had either vanish twin syndrome or had undergone fetal reduction for other reasons in one of their twins, and were subsequently subject to NIPT, chromosome karyotyping, chromosome microarray analysis (CMA), and short tandem repeat (STR) analysis. This study has been approved by the Medical Ethics Committee of Guangdong Maternal and Child Health Hospital (Ethics No.: 20230132). RESULTS: Case 1 underwent selective fetal reduction at 8⁺ weeks of gestation. At 17⁺ weeks, NIPT showed a fetal DNA fraction of 2.806%, with results indicating the presence of Y chromosome and abnormal sex chromosome ratios. However, the women had subsequent uncomplicated vaginal delivery of a female infant, and no abnormality noted. Case 2 experienced spontaneous demise of one twin at 13 weeks' gestation. At 19 weeks, NIPT indicated a high risk for chromosome 21 (Z-score 4.671) in the surviving fetus, but subsequent evaluation showed no abnormality. Case 3, a dichorionic diamniotic (DCDA) twin pregnancy, underwent selective reduction at 13⁺ weeks due to fetal abnormalities in one twin. At 22⁺ weeks, NIPT for the surviving fetus indicated a high risk for chromosome 21 (Z-score 17.549), but subsequent evaluation was unremarkable. CONCLUSION In twin pregnancies, the relatively low cell-free fetal DNA (cffDNA) concentration can compromise the success rate and accuracy of NIPT compared to singleton pregnancies. Residual DNA from the demised fetus may persist for weeks following VTS or selective reduction, potentially causing false-positive NIPT results and interfering with sex chromosome prediction for the surviving fetus. Additionally, determining chorionicity is critical for reliable interpretation of NIPT results in twin pregnancies.
Adult ; Female ; Humans ; Pregnancy ; Diseases in Twins/diagnosis* ; Karyotyping ; Noninvasive Prenatal Testing/methods* ; Pregnancy, Twin ; Prenatal Diagnosis/methods*

Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

Objective:To assess the preliminary efficacy and safety of modified Société Internationale d′Oncologie Pédiatrique Epithelial Liver Tumor Study Group (SIOPEL)-4 protocol for pediatric hepatoblastoma (HB) with lung metastases.Methods:This prospective cohort study enrolled 27 newly diagnosed pediatric HB with lung metastases who received the modified SIOPEL-4 protocol at Shanghai Children′s Medical Center, Shanghai Jiao Tong University School of Medicine, and Shanghai Children′s Hospital between January 2020 to December 2023. Clinical characteristics, lung response rates to induction chemotherapy, treatment outcomes, prognostic factors and sever chemotherapy toxicities at different stages were analyzed. Survival analysis was performed by Kaplan-Meier method. Univariate prognostic analysis was conducted by Log-Rank test.Results:Of the 27 patients, there were 17 males and 10 females, with the age of 21 (15, 33) months. During the follow-up of 31 (12, 45) months for 17 continuous complete remission patients, 4 cases disease progression (2 cases death) and 6 cases relapse were observed. The 2-year event free survival (EFS) and overall survival (OS) rate was (58±11)% and (89±7)%, respectively. All the 27 patients had response to block 1-3 induction chemotherapy (cisplatin+doxorubicin), with 14 cases (52%) achieving complete response and 13 cases (48%) achieving partial response of lung metastatic lesions, the 2-year EFS rate was (81±12)% and (34±14)%, respectively ( χ 2=6.76, P=0.009), the 2-year OS rate was 100% and (79±13)%, respectively ( χ2=2.12, P=0.145). Patients with caudate lobe tumors or ≥10 pulmonary metastatic nodules had significantly lower EFS rates ( χ2=5.36, 7.84, P=0.021, 0.005, respectively). The incidence of grade 3/4 neutropenia after block 1-3 induction chemotherapy, CD (carboplatin+doxorubicin), and VI (vincristine+irinotecan) consolidation chemotherapy was 90% (73/81), 75% (58/77), and 31% (11/35), respectively. The incidence of grade 3/4 thrombocytopenia was 77% (62/81), 69% (53/77), and 14% (5/35), respectively. The incidence of grade 3/4 infections was 64% (52/81), 25% (19/77), and 20% (7/35), respectively. The differences between the groups were statistically significant ( χ2=43.51, 42.69, 33.00, all P<0.001). Two patients (10%) of the 20 evaluable patients for ototoxicity occurred grade 3 and higher hearing impairment, with 1 patient requiring a hearing aid. Conclusions:The modified SIOPEL-4 regimen shows good preliminary efficacy and safety in treating pediatric HB with lung metastases. The prognosis for patients with residual lesions in the lungs after induction chemotherapy needs to be improved. Attention should be given to the ototoxicity induced by high-dose cisplatin chemotherapy.

BACKGROUND:In endodontics,revascularization and effective control of bacterial infection are prerequisite for regenerative repair of tissues and further development of the root apex.Ornidazole,carried in pulp-capping materials or vascularized scaffolding materials may control pulpal infections,but its effect on vascularization need to be investigated.OBJECTIVE:To investigate the residual concentration pattern of ornidazole in root canals and to evaluate the effects of ornidazole on endothelial cell proliferation,migration,and differentiation,as well as on vascular irritation.METHODS:(1)Ornidazole was encapsulated in the isolated pulp cavity and then immersed in Hank's balanced salt solution for 7 days.Ornidazole was then removed from the pulp cavity,reencapsulated in sterile water,and again immersed in Hank's balanced salt solution.The mass concentration of ornidazole in the pulp cavity fluid was measured periodically by colorimetric method.(2)Human umbilical vein endothelial cells were inoculated into well plates.Adherent cells were stimulated by the addition of lipopolysaccharide for 24 hours,and then co-cultured by the addition of 0,1,2,5,8,10 μg/mL ornidazole,to detect the cellular activity and migratory ability.Human umbilical vein endothelial cells were inoculated in well plates and co-cultured with different mass concentrations(0,1,2,5,8,10 μg/mL)of ornidazole or stimulated by lipopolysaccharide for 24 hours followed by the addition of different mass concentrations(0,1,2,5,8,10 μg/mL)of ornidazole.The gene expression of vascular endothelial growth factor and basic fibroblast growth factor as well as the protein expression of vascular endothelial growth factor was detected.(3)The chorioallantoic membrane assay was employed to assess the vascular irritation of 2 and 10 μg/mL ornidazole.RESULTS AND CONCLUSION:Residual ornidazole in exfoliated teeth was rapidly released within the initial 6 days,with a subsequent decrease in release rate,maintaining a concentration of approximately 2 μg/mL at the root apex after 8 days.Under lipopolysaccharide-induced inflammatory conditions,cell counting kit-8 and cell live-dead fluorescence staining showed that ornidazole(1-10 μg/mL)had no significant effect on the activity of human umbilical vein endothelial cells,and the cell scratch assay showed that ornidazole(1-10 μg/mL)had no obvious effect on the migratory ability of human umbilical vein endothelial cells.RT-qPCR assay showed that,after co-cultivation with ornidazole alone,the mRNA expression of vascular endothelial growth factor and basic fibroblast growth factor in human umbilical vein endothelial cells showed an overall decreasing trend.After co-culturing with ornidazole under lipopolysaccharide-induced inflammation,the mRNA expression of the two factors showed a rising trend in human umbilical vein endothelial cells.Western blot assay showed that vascular endothelial growth factor protein expression had an elevating trend in human umbilical vein endothelial cells after co-culture with ornidazole under lipopolysaccharide-induced inflammatory conditions.The chorioallantoic membrane assay showed that 2 and 10 μg/mL ornidazole were non-vascular irritating.To conclude,1-10 μg/mL ornidazole is non-cytotoxic and non-vascular irritating,promotes the expression of angiogenesis-related genes and proteins in inflammatory endothelial cells,and serves as a potential therapeutic agent for pulpal infection control.