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 retrospectively analyze the early monitoring indicators before and after admission, the use of coagulation/anticoagulant medications, and the red blood cell transfusion within 24 hours in emergency trauma patients, and to identify the indicators related to the volume of red blood cells transfused during the first 24 hours of hopitalization, thereby assisting clinical judgment of the probability and required quantity of red blood cell transfusion. Methods: Data of 117 emergency trauma patients admitted to intensive care unit (ICU) from January 2022 to March 2024 were retrospectively analyzed. Patients were categorized according to whether the volume of red blood cells transfused within 24 hours exceeded specific quartile thresholds (Q1, Q2, Q3). Non-parametric tests were used for numerical variables and Chi-square tests were used for categorical variables to identify statistically significant single-factor indicators, which were subsequently incoporated into a binary logistic regression model to obtain a combined predictive probability. ROC curve analysis was performed on the multi-factor indicators and their combined predictive probability derived from the binary logistic regression model. Results: 1) The initial hemoglobin (Hb) and hematocrit (Hct) levle were independent influencing factors in the group with red blood cell transfusion volume exceeded Q1 (P<0.05), and the combined predictive probability demostrated by ROC curve analysis was AUC=0.858 (P<0.05). 2) In the group of red blood cell transfusion volume exceeding Q2, the initial Fib, transhulitic acid, human prothrombin complex, trauma category and primary trauma site were independent influencing factors (P<0.05), and the combined predictive probability of ROC curve analysis was AUC=0.966 (P<0.05). 3) Pulse pressure and trauma category were independent influencing factors in the group with red blood cell transfusion volume exceeding Q3 (P<0.05), and ROC curve analysis revealed that combined prediction probability was AUC=0.944 (P<0.05). Conclusion: Early monitoring indicators and the use of coagulation medications, before and after admission in emergency trauma patients show diagnostic value in predicting the amount of red blood cells transfused on the first day of admission. Early warning alerts established through patient monitoring indicators can reduce incidents of untimely blood supply from the blood transfusion department (blood bank) for emergency trauma patients with massive hemorrhage, especially for patients with rare blood types or during blood supply shortage.

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 investigate the anti-inflammatory activity and potential mechanism of Clematis ranunculoides extract. METHODS The ear swelling was induced by xylene to establish an acute inflammation model of mice； using aspirin （0.25 g/kg） as a positive control， the effects of 1.25， 2.5， 5 g/kg C. ranunculoides extract on the degree of ear swelling were investigated. The chronic inflammation model of rats was also established by implanting cotton balls； using aspirin （0.17 g/kg） as a positive control， the effects of 0.88， 1.75， 3.5 g/kg C. ranunculoides extract on the net weight of granulomas were investigated. Furthermore， RAW264.7 cells were induced by lipopolysaccharide to establish an inflammatory injury model； the effects of 12.5， 25， 50 μg/mL C. ranunculoides extract on the contents of nitric oxide（NO）， prostaglandin E2（PGE2）， tumor necrosis factor-α （TNF-α）， interleukin-6（IL-6） and monocyte chemotactic protein-1（MCP-1） in the cell supernatant， the protein expressions of inducible nitric oxide synthase（iNOS）， cyclooxygenase-2（COX-2）， p65 and phosphorylated p65（p-p65） in cells as well as nuclear translocation of p65 protein were assessed. RESULTS C. ranunculoides extract with 5 g/kg significantly relieved ear swelling in mice， and C. ranunculoides extract with 1.75， 3.5 g/kg significantly decreased the net weight of granulomas in rats （P＜0.05）. C. ranunculoides extract with 12.5， 25， 50 μg/mL significantly reduced the contents of NO （except for 12.5 μg/mL C. ranunculoides extract）， PGE2， TNF-α， IL-6 and MCP-1 in the cell supernatant， as well as the relative expressions of iNOS and COX-2 protein， and relative expression ratio of p-p65 and p65 protein （P＜0.05 or P＜0.01）； C. ranunculoides extract with 25， 50 μg/mL inhibited the translocation of p65 protein to the cell nucleus. CONCLUSIONS C. ranunculoides extract exhibits significant anti- inflammatory activity， the mechanism of which may be attributed to the inhibition of the activation of nuclear factor-κB signaling pathway， down-regulation of COX-2 and iNOS protein expression， and the reduction of inflammatory cytokines release.

The continued aging of the population has led to a surge in long-term care needs for the elderly.Proactive preparation for future care(PFC)for the elderly can effectively meet the care needs caused by aging,improve their physical and mental health,and promote the elderly to maximize their physiological,psychological and social functions,finally realizing healthy aging.This paper reviews the theoretical basis,concept connotation,measurement tools,influencing factors and intervention strategies of PFC for the elderly,and puts forward the prospects for future research,so as to provide references for potential mechanism and empirical research of PFC for Chinese seniors.

The orbitofrontal cortex (ORB), a region crucial for stimulus-reward association, decision-making, and flexible behaviors, extensively connects with other brain areas. However, brain-wide inputs to projection-defined ORB neurons and the distribution of inhibitory neurons postsynaptic to neurons in specific ORB subregions remain poorly characterized. Here we mapped the inputs of five types of projection-specific ORB neurons and ORB outputs to two types of inhibitory neurons. We found that different projection-defined ORB neurons received inputs from similar cortical and thalamic regions, albeit with quantitative variations, particularly in somatomotor areas and medial groups of the dorsal thalamus. By counting parvalbumin (PV) or somatostatin (SST) interneurons innervated by neurons in specific ORB subregions, we found a higher fraction of PV neurons in sensory cortices and a higher fraction of SST neurons in subcortical regions targeted by medial ORB neurons. These results provide insights into understanding and investigating the function of specific ORB neurons.
Animals ; Neurons/physiology* ; Mice ; Prefrontal Cortex/cytology* ; Parvalbumins/metabolism* ; Brain Mapping/methods* ; Neural Pathways/physiology* ; Somatostatin/metabolism* ; Male ; Interneurons/physiology* ; Mice, Inbred C57BL ; Thalamus/physiology* ; Mice, Transgenic

The secondary motor cortex (M2) encodes choice-related information and plays an important role in cue-guided actions. M2 neurons innervate the dorsal striatum (DS), which also contributes to decision-making behavior, yet how M2 modulates signals in the DS to influence perceptual decision-making is unclear. Using mice performing a visual Go/No-Go task, we showed that inactivating M2 projections to the DS impaired performance by increasing the false alarm (FA) rate to the reward-irrelevant No-Go stimulus. The choice signal of M2 neurons correlated with behavioral performance, and the inactivation of M2 neurons projecting to the DS reduced the choice signal in the DS. By measuring and manipulating the responses of direct or indirect pathway striatal neurons defined by M2 inputs, we found that the indirect pathway neurons exhibited a shorter response latency to the No-Go stimulus, and inactivating their early responses increased the FA rate. These results demonstrate that the M2-to-DS pathway is crucial for suppressing inappropriate responses in perceptual decision behavior.
Mice ; Animals ; Motor Cortex ; Corpus Striatum/physiology* ; Neostriatum ; Neurons/physiology* ; Reaction Time

Objective:To assess the ultrasonographic features and potential diseases of fetal abnormal sylvian fissure(SF), and to explore the value of whole-genome sequencing (WGS) in prenatal detection.Methods:A total of 28 fetuses with a sonographic diagnosis of abnormal SF in Shenzhen Maternal and Child Health Hospital Affiliated to Southern Medical University between October 2018 and October 2020 were prospectively included. The fetal brain was evaluated by neuroultrasound and intrauterine MRI in detail. Amniotic fluid/cord blood obtained by amniocentesis or tissue samples from umbilical cord after birth were collected for WGS. Pregnancy outcomes and postnatal MRI were recorded, and neurodevelopment of live-born infants was followed up for more than 24 months after delivery.Results:During the study period, 28 fetuses with abnormal SF were identified, with a gestational age of 21.3-30.0 (24.8±2.0) weeks. Abnormal SF presented in MCD ( n=15, 53.6%), chromosomal anomalies ( n=3, 10.7%) or single-gene genetic syndromes ( n=3, 10.7%) with the affected fetuses showing developmental delay, hydrocephalus or leukomalacia ( n=4, 14.2%), corpus callosal agenesis with large interhemispheric cysts ( n=1, 3.6%), benign subarachnoid space enlargement with arachnoid cysts ( n=1, 3.6%), and multiple malformations ( n=1, 3.6%). Among the 15 cases with MCD, the most common pathology was lissencephaly/pachygyria, followed by schizencephaly, severe microcephaly, hemimegalencephaly with paraventricular heterotopia, and polymicrogyria. Abnormal SF presented bilaterally in 23 fetuses and unilaterally in 5. All cases were categorized into six types depending on SF morphology in the transthalamic section: no plateau-like or a small insula, linear type, irregular corrugated SF, Z-shaped, and cyst occupying type. In addition to abnormal SF, associated anomalies or mild variations were identified in all fetuses. There were 17 cases underwent intrauterine MRI, and 13 cases underwent postnatal MRI examination.And 25 pregnancies were terminated; 3 were born alive, and 2 had typical syndromic changes with poor neurodevelopmental prognosis. A related pathogenic genetic variant was detected in 57.1% (16/28) fetus, and the incidence of single nucleotide variants(SNVs) was 42.9% (12/28), among which de novo SNVs accounted for 91.7% (11/12). Conclusions:Fetal abnormal SF could be classified based on the ultrasonographic features of transthalamic section. Fetal abnormal SF may indicate MCD, some chromosomal abnormalities or single-gene genetic syndromes that may lead to poor neurodevelopmental outcomes, and may be affected by extra-cortical factors. It is suggested to carry out targeted prenatal genetic diagnosis for fetuses with abnormal SF.

Objective:To explore the feasibility of deep learning-based restoration of obscured thyroid ultrasound images.Methods:A total of 358 images of thyroid nodules were retropectively collected from January 2020 to October 2021 at Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, and the images were randomly masked and restored using DeepFillv2. The difference in grey values between the images before and after restoration was compared, and 6 sonographers (2 chief physicians, 2 attending physicians, 2 residents) were invited to compare the rate of correctness of judgement and detection of image discrepancies. The ultrasound features of thyroid nodules (solid composition, microcalcifications, markedly hypoechoic, ill-defined or irregular margins, or extrathyroidal extensions, vertical orientation and comet-tail artifact) were extracted according to the Chinese Thyroid Imaging Reporting and Data System (C-TIRADS). The consistency of ultrasound features of thyroid nodules before and after restoration were compared.Results:The mean squared error of the images before and after restoration ranged from 0.274 to 0.522, and there were significant differences in the rate of correctness of judgement and detection of image discrepancies between physicians of different groups(all P<0.001). The overall accuracy rate was 51.95%, the overall detection rate was 1.79%, there were significant differences also within the chief physicians and resident groups (all P<0.001). The agreement rate of all ultrasound features of the nodules before and after image restoration was higher than 70%, over 90% agreement rate for features such as solid composition and comet-tail artifact. Conclusions:The algorithm can effectively repair obscured thyroid ultrasound images while preserving image features, which is expected to expand the deep learning image database, and promote the development of deep learning in the field of ultrasound images.

【Objective】 To investigate the predictive value of nutritional risk index (NRI), systemic immune inflammatory index (SⅡ) and triglyceride glucose (TyG) index on the condition and prognosis of patients with acute pancreatitis (AP). 【Methods】 A total of 173 AP patients were divided into mild acute pancreatitis (MAP) group (n=79), moderate acute pancreatitis (MSAP) group (n=44), and severe acute pancreatitis (SAP) group (n=50) according to their severity. All the 50 SAP patients were divided into death group (19 cases) and survival group (31 cases) according to the death situation. The NRI, SⅡ and TyG indexes of each group were recorded and compared. The values of NRI, SⅡ and TyG index in predicting the occurrence and death of SAP were analyzed with ROC curve. Pearson correlation analysis of the correlation between NRI, SⅡ, and TyG index in SAP patients was made. 【Results】 NRI was significantly lower in SAP group (89.25±4.50) than in MSAP group (93.40±6.25) and MAP group (97.62±8.60), while SⅡand TyG index in SAP group (2 706.30±1 052.74, 7.84±1.21) were significantly higher than those in MSAP group (1 937.24±983.48, 6.52±1.05) and MAP group (1 280.58±717.36, 4.65±0.58) (P<0.001). NRI in death group (86.40±3.70) was significantly lower than that in survival group (91.46±5.28), while SⅡ and TyG index in death group (3 085.73±1 192.48, 9.05±1.37) were significantly higher than those in survival group (2 270.26±994.53, 6.70±1.10) (P<0.001). The ROC curve showed that the AUC of NRI, SⅡ and TyG index jointly predicting SAP occurrence and death was 0.850 (95% CI: 0.792-0.908) and 0.905 (95% CI: 0.843-0.966), respectively. Correlation analysis showed that NRI was negatively correlated with SⅡ and TyG index in SAP patients (r=-0.761, P<0.001, r=-0.813, P<0.001), while SⅡ was positively correlated with TyG index (r=0.842, P<0.001). 【Conclusion】 NRI, SⅡ and TyG index are related to the severity and death of AP patients, and the combination of the three indexes has good value in predicting the occurrence and prognosis of SAP.