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.

Humans ; Acute Disease ; Pancreatitis/diagnosis* ; Follow-Up Studies ; Patient Discharge ; Disease Progression

ObjectiveBased on cyclic adenosine monophosphate （cAMP）/cAMP-regulated guanine nucleotide exchange factor 1 （Epac1）/Ras-homologous protein 1 （Rap1） signaling pathway to explore the myocardial protective mechanism of Yiqi Huoxue prescription on coronary heart disease （CHD） rats with Qi deficiency and blood stasis syndrome. MethodEighty-eight specific-pathogen-free （SPF） grade male Sprague-Dawley （SD） rats were divided into a sham operation group （n=12） and an experimental group （n=76） according to the random number table. The experimental group underwent a restricted diet and exhaustive swimming combined with left anterior descending （LAD） coronary artery ligation to construct a model of CHD with Qi deficiency and blood stasis syndrome， and electrocardiograms （ECGs） of rats before and after the LAD operation were collected. After the model was successfully established， the rats were randomly divided into model group， Yiqi Huoxue prescription low-dose group （4.28 g·kg^-1）， medium-dose group （8.55 g·kg^-1）， high-dose group （17.1 g·kg^-1）， and Western medicine group （isosorbide mononitrate tablets， 3.6 mg·kg^-1）. Rats were intragastrically administered assigned drugs for 4 weeks consecutively， while the sham operation group and the model group were administered with equal volumes of double distilled water. Twenty-four hours after the final administration， the rats were anesthetized with isoflurane to detect the left ventricular end-diastolic diameter （LVEDD）， left ventricular end-systolic diameter （LVESD）， left ventricular fractional shortening （FS， %）， and ejection fraction （EF， %） by echocardiography. Blood samples were collected from the abdominal aorta for hemorheological measurements， and plasma cAMP levels were determined using enzyme-linked immunosorbent assay （ELISA）. Myocardial tissue was collected for hematoxylin-eosin （HE） staining and Masson staining to observe myocardial pathological damage， and a transmission electron microscope was used to observe ultrastructural changes of myocardial tissue， fluorescent quantitative real-time polymerase chain reaction （Real-time PCR） was used to detect the expressions of myocardial Epac1， Rap1 GTPase activating protein （Rap1GAP） and Rap1 mRNA， and Western blot was used to detect the expressions of myocardial Epac1， Rap1GAP and Rap1 protein. ResultCompared with those in the sham operation group， LVEDD and LVISD of rats in the model group significantly increased （P<0.01）， and the ratios of EF and FS significantly decreased （P<0.01）， indicating symptoms of heart function decline， referred to as "heart Qi deficiency". The viscosity of whole blood and plasma significantly increased （P<0.01）， and the content of cAMP significant increased （P<0.01）. In addition， there was a significant proliferation of collagen fibers in myocardial tissue （P<0.01）， and the ultrastructure of the myocardial tissue was severely damaged， indicating pathological changes consistent with "blood stasis". Real-time PCR results showed that Epac1 and Rap1 mRNA levels in the model group were significantly reduced （P<0.01）， while Rap1GAP mRNA levels were significantly increased （P<0.01）. Western blot analysis showed a significant decrease in Epac1 protein expression （P<0.01） and a significant increase in Rap1GAP protein expression （P<0.05）. Compared with the model group， Yiqi Huoxue prescription improved cardiac function， reduced blood viscosity， lowered plasma cAMP levels， decreased collagen fiber proliferation， and improved myocardial ultrastructure damage in CHD rats with Qi deficiency and blood stasis syndrome. The high-dose group showed the most significant effects. In the high-dose group， Epac1 mRNA and protein expression levels significantly increased （P<0.01）， Rap1 mRNA expression significantly increased （P<0.01）， and Rap1GAP mRNA and Rap1GAP/Rap1 protein expression levels significantly decreased （P<0.05， P<0.01）. ConclusionYiqi Huoxue prescription can improve cardiac function， reduce blood viscosity and plasma cAMP levels， improve myocardial damage， and reduce collagen fiber proliferation in CHD rats with Qi deficiency and blood stasis syndrome. The myocardial protection mechanism may be related to the regulation of the cAMP/Epac1/Rap1 signaling pathway.

Objective:To investigate the effect of the number of positive preoperative serological tumor markers on the surgical approach and prognosis of patients with intrahepatic cholangiocarcinoma.Methods:This is a retrospective case-series study. Data from 548 patients with intrahepatic cholangiocarcinoma after radical resection from October 2010 to April 2019 were retrospectively collected in 10 hospitals of China. There were 277 males and 271 females with an age of (57.8±10.2)years(range:23 to 84 years). Four hundred and twenty-six patients(77.7%) had at least one positive preoperative serum tumor marker. The data collection included the results of 4 preoperative serological tumor markers,other preoperative indicators(5 prodromal symptoms, 6 medical history,8 preoperative serological indicators,5 preoperative imaging indicators,and 14 preoperative pathological examination indicators),baseline data (gender and age),surgical methods,and prognostic follow-up data. Four preoperative results of serologic tumor marker and surgical procedure were converted into categorical variables. The number of positive preoperative serum tumor markers was used as the treatment variable,the surgical method was used as the mediating variable,and the survival time was used as the outcome variable. Univariate and multivariate analysis were used to screen for other preoperative indicators which were independent factors that influenced the surgical procedure and the prognosis of patients as covariates to analyze the mediating effect.Results:Of the 548 patients included in the study, 176 patients (32.1%) underwent partial hepatectomy,151 patients(27.5%) underwent hemihepatectomy, and 221 patients(40.3%) underwent partial hepatectomy or hemihepatectomy combined with other treatments. The results of the univariate and multivariate analysis showed that the number of positive serum tumor markers,intrahepatic bile duct dilatation,portal vein invasion,pathological differentiation,pathological type,vascular invasion,T stage,N stage and maximum tumor diameter were independent factors influencing the surgical procedure(all P<0.05). Intrahepatic bile duct dilatation,pathological differentiation and T stage were independent prognostic factors for patients with intrahepatic cholangiocarcinoma(all P<0.05). Intrahepatic bile duct dilatation,differentiation and T stage were included as covariates in the mediation effect model. The results showed that the number of positive serum tumor markers before surgery had a negative predictive effect on the survival time of patients with intrahepatic cholangiocarcinoma ( β=-0.092, P=0.039),and had a positive predictive effect on the surgical method ( β=0.244, P<0.01). The number of positive serum tumor markers had a negative predictive effect on the survival time of patients with intrahepatic cholangiocarcinoma ( β=-0.151, P=0.002). Direct and indirect effects accounted for 71.3% and 28.7% of total effects,respectively. Conclusions:The higher the positive number of preoperative tumor markers,the worse the prognosis of patients with intrahepatic cholangiocarcinoma. The number of positive cells not only directly affects the prognosis of patients,but also indirectly affects the prognosis of patients by affecting the surgical method.

Objective:To explore the repairing effect and mechanism of tangeretin in rats with spinal cord injury.Methods:The rats were divided into sham-operation group, model group and tangeretin group according to random number table, with 8 rats in each group. Except for the sham-operation group, Allen hit method was used to make rat models in the other groups. After the model was successfully established, the tangeretin group was intragastrically administered with tangeretin 50 mg/kg, and the sham-operation group and the model group were intragastrically administered with an equal volume of normal saline once a day for 14 days. On days 0, 3, 7, and 14 after modeling, the motor function recovery of rats was assessed using the Basso-Beattie-Bresnahan (BBB) score; the morphological changes of the spinal cord tissues were observed using HE staining and Nissl staining; the SOD and GSH activities and MDA, IL-1β, TNF-α, and IL-10 levels in the spinal cord tissues of rats in each group were measured using ELISA kit detection; the GFAP and Neun expressions in the spinal cord tissues were detected by immunofluorescence; the IL-1β, TNF-α, IL-10, nuclear factor E2-related factor 2 (Nrf-2), and NAD (P) H-quinone oxidoreductase 1 (NQO-1) expressions in the spinal cord tissues were detected by Western blot.Results:Compared with the model group, the BBB score increased in the tangeretin group ( P<0.05), HE staining score decreased ( P<0.05), and the number of Nissl bodies increased ( P<0.05); the level of IL-10, SOD and GSH activities increased ( P<0.05), and IL-1β, TNF-α and MDA levels decreased in the spinal cord tissue ( P<0.05); GFAP fluorescence intensity decreased ( P<0.05) and NeuN fluorescence intensity increased ( P<0.05); the relative expression of IL-1β and TNF-α decreased ( P<0.05), and the relative expressions of IL-10, Nrf-2 and NQO-1 protein increased ( P<0.05). Conclusions:Tangeretin can exert anti-inflammatory and anti-oxidative stress effects through the Nrf2/NQO1 signaling pathway and alleviate early spinal cord injury in rats. On the other hand, it may promote the recovery of spinal cord injury by reducing glial scar generation and promoting neural cellogenesis.

Objective To investigate the effect of varying blood pressure stratification on renal function in the diabetic population.Methods A prospective cohort study was conducted,enrolling 9 489 diabetic patients from a total of 101 510 Kailuan Group employees who underwent health examinations between July 2006 and October 2007.The follow-up period was(8.6±4.0)years.Participants were categorized into four groups based on their baseline blood pressure levels:normal blood pressure(systolic blood pressure<120 mmHg and diastolic blood pressure<80 mmHg),elevated blood pressure(systolic blood pressure 120-130 mmHg and diastolic blood pressure<80 mmHg),stage 1 hypertension(systolic blood pressure 130-140 mmHg and/or diastolic blood pressure 80-90 mmHg),and stage 2 hypertension(systolic blood pressure≥140 mmHg and/or diastolic blood pressure≥90 mmHg).The incidence density of chronic kidney disease(CKD)was compared among these groups.A multivariate Cox proportional hazards regression model was employed to assess the effects of different blood pressure levels on renal function in diabetic patients,with the stability of the results confirmed using a multivariate time-dependent Cox proportional hazards model.Sensitivity analysis was conducted after excluding cases of cardiovascular disease(CVD)during follow-up,and cases using antihypertensive and antidiabetic medications at baseline.Results(1)At baseline,stage 1 hypertension patients demonstrated statistically significant higher differences with age and body mass index(BMI)compared to normal blood pressure group(P<0.05).(2)By the end of the follow-up,2 294 cases of CKD were identified,including 1 117 cases of estimated glomerular filtration rate(eGFR)decline and 1 575 cases of urinary protein.The incidences density of CKD,eGFR decline and urinary protein for stage 1 hypertension group were 39.4,16.3 and 25.5 per thousand person-years,respectively,all of which were statistically significant different from normal blood pressure group(log-rank test,P<0.01).(3)Multivariate Cox regression analysis revealed that,compared to the normal blood pressure group,stage 1 hypertension was associated with a 29%increased risk of CKD(HR=1.29,95%CI 1.09-1.52)and a 40%increased risk of eGFR decline(HR=1.40,95%CI 1.08-1.80)in diabetic individuals.Conclusion Stage 1 hypertension significantly increases the risk of CKD and eGFR decline in diabetic individuals,with a particularly notable effect on the risk of eGFR decline.

Objective To explore the effects of 1α,25-dihydroxyvitamin D3 on airway inflammation in asthmatic mice and the potential mechanisms.Methods Twenty-four female BALB/c mice in SPF grade were randomly divided into three groups(n=8):control group,asthma group,and asthma+VD3 group.On the 1st,8th,and 15th day,asthma group and asthma+VD3 group were given 0.2 ml ovalbumin(OVA)suspension for sensitization,while control group received 0.2 ml normal saline.On the 22-28th day,asthma group and asthma+VD3 group were challenged with 1%OVA atomization inhalation,while control group received an equal amount of normal saline atomization,for 30 minutes each time,once a day,for a continuous 7 days.Asthma+VD3 group was given intraperitoneal injection of 1α,25-dihydroxyvitamin D3 injection(4 μg/kg)30 minutes before each atomization,while control group and asthma group were given an equal dose of normal saline.After the last challenge,all mice were anesthetized,and serum,bronchoalveolar lavage fluid(BALF)and lung tissue samples were collected.HE staining and Periodic Acid Schiff(PAS)staining were used to observe the pathological changes in lung tissue and changes in airway mucus levels.ELISA was employed to detect serum IgE and inflammatory cytokines IL-4,IL-5 and IL-13 in BALF.Immunohistochemical technique and Western blotting were used to detect the expressions of SIRT1 and GATA-3 in mouse lung tissue.Results Compared with control group,asthma group had a significant increase in inflammatory cell infiltration around lung tissue,bronchia and accompanying perivascular,mainly characterized by eosinophils.Bronchial lumen stenosis,airway mucosal epithelial hyperplasia,and increased tracheal mucus secretion were also observed.The above changes in asthma+VD3 group were reduced compared with asthma group.Compared with control group,serum levels of IgE,and IL-4,IL-5,IL-13 inflammatory factors in BALF and GATA-3 in lung tissue were increased in asthma group(P<0.05),and SIRT1 level in lung tissue was significant decreased(P<0.05).Compared with asthma group,IgE level in serum,inflammatory factors of IL-4,IL-5 and IL-13 in BALF,and GATA-3 in lung tissue in asthma+VD3 group were decreased(P<0.05),and SIRT1 level in lung tissue was increased(P<0.05).Correlation analysis showed that the expression level of lung tissue SIRT1 was negatively correlated with the expression of GATA-3,serum IgG,and the levels of IL-4,IL-5,and IL-13 in BALF(P<0.05);the expression level of lung tissue GATA-3 was positively correlated with serum IgG and the levels of IL-4,IL-5,and IL-13 in BALF(P<0.05).Conclusion 1α,25-dihydroxyvitamin D3 can alleviate airway inflammation in asthmatic mice,possibly by upregulating the expression of SIRT1 in lung tissue and inhibiting the expression of GATA-3,thereby inhibiting inflammatory factors(IL-4,IL-5,IL-13).

Objective:To investigate the effect of the number of positive preoperative serological tumor markers on the surgical approach and prognosis of patients with intrahepatic cholangiocarcinoma.Methods:This is a retrospective case-series study. Data from 548 patients with intrahepatic cholangiocarcinoma after radical resection from October 2010 to April 2019 were retrospectively collected in 10 hospitals of China. There were 277 males and 271 females with an age of (57.8±10.2)years(range:23 to 84 years). Four hundred and twenty-six patients(77.7%) had at least one positive preoperative serum tumor marker. The data collection included the results of 4 preoperative serological tumor markers,other preoperative indicators(5 prodromal symptoms, 6 medical history,8 preoperative serological indicators,5 preoperative imaging indicators,and 14 preoperative pathological examination indicators),baseline data (gender and age),surgical methods,and prognostic follow-up data. Four preoperative results of serologic tumor marker and surgical procedure were converted into categorical variables. The number of positive preoperative serum tumor markers was used as the treatment variable,the surgical method was used as the mediating variable,and the survival time was used as the outcome variable. Univariate and multivariate analysis were used to screen for other preoperative indicators which were independent factors that influenced the surgical procedure and the prognosis of patients as covariates to analyze the mediating effect.Results:Of the 548 patients included in the study, 176 patients (32.1%) underwent partial hepatectomy,151 patients(27.5%) underwent hemihepatectomy, and 221 patients(40.3%) underwent partial hepatectomy or hemihepatectomy combined with other treatments. The results of the univariate and multivariate analysis showed that the number of positive serum tumor markers,intrahepatic bile duct dilatation,portal vein invasion,pathological differentiation,pathological type,vascular invasion,T stage,N stage and maximum tumor diameter were independent factors influencing the surgical procedure(all P<0.05). Intrahepatic bile duct dilatation,pathological differentiation and T stage were independent prognostic factors for patients with intrahepatic cholangiocarcinoma(all P<0.05). Intrahepatic bile duct dilatation,differentiation and T stage were included as covariates in the mediation effect model. The results showed that the number of positive serum tumor markers before surgery had a negative predictive effect on the survival time of patients with intrahepatic cholangiocarcinoma ( β=-0.092, P=0.039),and had a positive predictive effect on the surgical method ( β=0.244, P<0.01). The number of positive serum tumor markers had a negative predictive effect on the survival time of patients with intrahepatic cholangiocarcinoma ( β=-0.151, P=0.002). Direct and indirect effects accounted for 71.3% and 28.7% of total effects,respectively. Conclusions:The higher the positive number of preoperative tumor markers,the worse the prognosis of patients with intrahepatic cholangiocarcinoma. The number of positive cells not only directly affects the prognosis of patients,but also indirectly affects the prognosis of patients by affecting the surgical method.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate the value of quantitative ultrashort-echo-time (UTE) adiabatic T 1ρ (AdiabT 1ρ) in the diagnosis of articular cartilage degeneration of knee. Methods:A prospective case-control study was used. From March 2023 to September 2024, 30 healthy volunteers and 50 patients with osteoarthritis (OA) were recruited in the Second Affiliated Hospital, South China University of Technology and the Fifth Affiliated Hospital of Sun Yat-sen University. The unilateral knee joints were scanned by using three-dimensional (3D) UTE-Cones-AdiabT 1ρ sequence. The articular cartilages of knee were divided into 13 subregions slice by slice, with a total of 3 837 subregions. The whole-organ magnetic resonance-imaging score (WORMS) was evaluated in each subregion. According to the extent of lesions, the subregions cartilages were divided into normal group (WORMS 0, 2 589 subregions), localized lesion group (WORMS 1, 2, 2.5, 911 subregions) and diffuse lesion group (WORMS 3, 4, 5, 337 subregions). According to the depth of lesions, they were divided into normal group (WORMS 0, 2 589 subregions), partial cartilage layer lesions group (WORMS 1, 2, 3, 4, 1 108 subregions) and full-thickness lesion group (WORMS 2.5, 5, 140 subregions). WORMS 1 was early cartilage degeneration. The Kellgren-Lawrence (KL) grade of the knee joints was assessed and divided into normal group (KL grade 0, 1 323 subregions), suspicious or mild OA group (KL grades 1 and 2, 1 738 subregions), and moderate or severe OA group (KL grades 3 and 4, 776 subregions). The UTE-Cones-AdiabT 1ρ values and curves for each subregion were obtained by Matlab 2017b software. The ANOVA was used to compare the differences in UTE-Cones-AdiabT 1ρ between different WORMS groups and KL groups. Spearman correlation analysis was used to evaluate the correlation between UTE-Cones-AdiabT 1ρ and WORMS and KL groups. The receiver operating characteristic curve and area under the curve (AUC) were used to determine the efficacy of UTE-Cones-AdiabT 1ρ in diagnosing early cartilage degeneration and suspicious or mild OA. Results:Both normal volunteers and OA patients were able to obtain good cartilage images and well-fitting UTE-Cones-AdiabT 1ρ curves. There were 2 589, 569, 277, 65, 199, 63 and 75 cartilage subregions with WORMS scores of 0, 1, 2, 2.5, 3, 4 and 5, respectively, and the UTE-Cones-AdiabT 1ρ values were (36.1±5.1), (41.5±5.2), (42.3±5.3), (45.2±6.3), (43.0±5.2), (44.1±5.4) and (47.5±5.7) ms, respectively. The difference was statistically significant ( F=161.63, P<0.001). There was a statistically significant difference in UTE-Cones-AdiabT 1ρ among normal group, localized lesion group and diffuse lesion group ( F=450.92, P<0.001), and there was a significant difference in UTE-Cones-AdiabT 1ρ value among normal group, partial cartilage lesion group and full-thickness lesion group ( F=479.13, P<0.001). There was a statistically significant difference in UTE-Cones-AdiabT 1ρ among the normal group, the suspected or mild OA group, and the moderate or severe OA group ( F=36.91, P<0.001). The UTE-Cones-AdiabT 1ρ values in all subregions were positively correlated with WORMS and KL grades ( r=0.55, P<0.001; r=0.26, P<0.001). The AUC of UTE-Cones-AdiabT 1ρ in the diagnosis of early cartilage degeneration was 0.773 (95% CI 0.750-0.796), and the AUC in the diagnosis of suspicious or mild OA was 0.580 (95% CI 0.560-0.601). Conclusion:3D UTE-Cones-AdiabT 1ρ is positively correlated with the degree of cartilage degeneration, which can quantitatively evaluate cartilage degeneration and has the value of early diagnosis of articular cartilage degeneration.