BACKGROUND:Overactive osteoclasts disrupt bone homeostasis and play a bad role in the pathological mechanisms of related skeletal diseases,such as osteoporosis,fragility fractures,and osteoarthritis.Studies have confirmed that ellagic acid and ellagtannin have the potential to inhibit osteoclast differentiation.As their natural metabolites,urolithin A has antioxidant,anti-inflammatory,anti-proliferative and anti-cancer effects,but its effect on osteoclast differentiation and its underlying molecular mechanisms remain unclear. OBJECTIVE:To explore the effect of urolithin A on osteoclast differentiation induced by receptor activator for nuclear factor-κB ligand and its mechanism. METHODS:Mouse mononuclear macrophage leukemia cells(RAW264.7)that grew stably were cultured in vitro.Toxicity of urolithin A(0,0.1,0.5,1.5,2.5 μmol/L)to RAW264.7 cells were detected by cytotoxic MTS assay to screen out the safe concentration.Different concentrations of urolithin A were used again to intervene with receptor activator for nuclear factor-κB ligand-induced differentiation of RAW264.7 cells in vitro.Then,tartrate-resistant acid phosphatase staining and F-actin ring and nucleus staining were performed to observe its effect on the formation and function of osteoclasts.Finally,the expressions of urolithin A on upstream and downstream genes and proteins in the MAPK signaling pathway were observed by western blot and RT-qPCR assays. RESULTS AND CONCLUSION:Urolithin A inhibited osteoclast differentiation and F-actin ring formation in a concentration-dependent manner and 2.5 μmol/L had the strongest inhibitory effect.Urolithin A inhibited the mRNA expression of Nfatc1,Ctsk,Mmp9 and Atp6v0d2 and the protein synthesis of Nfatc1 and Ctsk,related to osteoclast formation and bone resorption.Urolithin A inhibited the activity of osteoclasts by downregulating the phosphorylation of p38 protein to inhibit the mitogen-activated protein kinase signaling pathway.

Objective:To conduct evidence-based exercise training for patients with peripheral arterial disease, develop review indicators, analyze barriers and enablers in the evidence-based practice process, and develop change strategies.Methods:Guided by the clinical evidence application model of the Joanna Briggs Institute Evidence-Based Health Care Center, the study identified clinical nursing problems, formed the evidence-based practice team, systematically searched, evaluated, and summarized evidence of exercise training in patients with peripheral arterial disease, developed review indicators, and clarified review methods. The baseline review was conducted from October 1 to 31, 2022. The integrated-promoting action on research implementation in health services (i-PARIHS) framework was used to analyze the barriers and enablers of the baseline review results, and corresponding strategies were formulated.Results:A total of 20 best evidence were included, and 11 review indicators were developed, with only one indicator having a compliance rate of 100%. This study analyzed 22 barriers and 24 enablers, and formulated 14 change strategies.Conclusions:The review indicators constructed based on the best evidence are scientific, effective, appropriate, and feasible. The analysis of barriers and enablers, as well as the formulation of change strategies, can provide guarantees for promoting clinical practice of exercise training for patients with peripheral arterial diseases.

Background::We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) enhances their remuscularization capacity after human cardiac muscle patch transplantation in infarcted mouse hearts. Herein, we sought to identify the effect of magnesium lithospermate B (MLB) on hiPSC-CMs during myocardial repair using a myocardial infarction (MI) mouse model.Methods::In C57BL/6 mice, MI was surgically induced by ligating the left anterior descending coronary artery. The mice were randomly divided into five groups ( n = 10 per group); a MI group (treated with phosphate-buffered saline only), a hiPSC-CMs group, a MLB group, a hiPSC-CMs + MLB group, and a Sham operation group. Cardiac function and MLB therapeutic efficacy were evaluated by echocardiography and histochemical staining 4 weeks after surgery. To identify the associated mechanism, nuclear factor (NF)-κB p65 and intercellular cell adhesion molecule-1 (ICAM1) signals, cell adhesion ability, generation of reactive oxygen species, and rates of apoptosis were detected in human umbilical vein endothelial cells (HUVECs) and hiPSC-CMs. Results::After 4 weeks of transplantation, the number of cells that engrafted in the hiPSC-CMs + MLB group was about five times higher than those in the hiPSC-CMs group. Additionally, MLB treatment significantly reduced tohoku hospital pediatrics-1 (THP-1) cell adhesion, ICAM1 expression, NF-κB nuclear translocation, reactive oxygen species production, NF-κB p65 phosphorylation, and cell apoptosis in HUVECs cultured under hypoxia. Similarly, treatment with MLB significantly inhibited the apoptosis of hiPSC-CMs via enhancing signal transducer and activator of transcription 3 (STAT3) phosphorylation and B-cell lymphoma-2 (BCL2) expression, promoting STAT3 nuclear translocation, and downregulating BCL2-Associated X, dual specificity phosphatase 2 (DUSP2), and cleaved-caspase-3 expression under hypoxia. Furthermore, MLB significantly suppressed the production of malondialdehyde and lactate dehydrogenase and the reduction in glutathione content induced by hypoxia in both HUVECs and hiPSC-CMs in vitro. Conclusions::MLB significantly enhanced the potential of hiPSC-CMs in repairing injured myocardium by improving endothelial cell function via the NF-κB/ICAM1 pathway and inhibiting hiPSC-CMs apoptosis via the DUSP2/STAT3 pathway.

COVID-19 is caused by a novel coronavirus-severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), which has being spreading around the world, posing a serious threat to human health and lives. Neutralizing antibodies and small molecule inhibitors for virus replication cycle are the main antiviral treatment for novel coronavirus recommended in China. To further promote the rational use of antiviral therapy in clinical practice, the National Center for Infectious Diseases (Beijing Ditan Hospital Capital Medical University and the First Affiliated Hospital, Zhejiang University School of Medicine) invited experts in fields of infectious diseases, respiratory and intensive care to develop an Expert Consensus on Antiviral Therapy of COVID-19 based on the Diagnosis and Treatment Guideline for COVID-19 ( trial version 10) and experiences in the diagnosis and treatment of COVID-19 in China. The consensus is concise, practical and highly operable, hopefully it would improve the understanding of antiviral therapy for clinicians and provide suggestions for standardized medication in treatment of COVID-19.

Objective:A high performance liquid chromatography (HPLC) method was developed to determine the enzymatic activity of CD38 in blood, which was the major enzyme responsible for consuming nicotinamide adenine dinucleotide (NAD). Additionally, the study aimed to detect the differences in CD38 enzymatic activity among individuals of varying ages and health statuses.Methods:A 50 μl whole blood matrix and enzyme reaction substrate of 150 μl β-NAD at a concentration of 500 μmol/L were selected for the analysis. To eliminate the impact of endogenous β-NAD, the whole blood sample was pre-incubated at 37 ℃ for 20 minutes before adding the substrate. The reaction was terminated by perchloric acid (PCA) after incubation at 37 ℃ for 40 min. The change in product nicotinamide (NAM) before and after the enzymatic reaction was measured by HPLC to calculate the CD38 activity. The linearity, limit of detection, limit of quantification, precision, and stability of the method were evaluated. The CD38 enzymatic activities in 60 healthy volunteers and 30 colorectal cancer patients in blood were determined by the developed method.Results:Pre-incubation at 37 ℃ for 20 minutes eliminated the effect of endogenous β-NAD. The correlation coefficient of NAM was 0.999 in the concentration range of 0.1-3.2 μmol/L, with limit of detection of 0.5 nmol/L and limit of quantification of 2.1 nmol/L. The average within-run imprecision ( CV) and total CV were 3.22%-4.03% and 2.91%-4.70%, respectively. The recovery rate ranged from 94.82% to 96.81%. The CD38 activity of whole blood was stable by storage at 4 ℃ for 48 hours, storage at room temperature for 8 hours, thawing of frozen whole blood at room temperature for 2 hours, or repeated freeze-thawing three times. NAM, NAD standards, and pre-treatment samples were stable after 48 hours at 4 ℃ and 8 hours at room temperature. CD38 activity gradually decreased with increasing concentration of the added CD38 inhibitor 4-aminoquinoline derivative (78c). Measurement of 60 healthy physical examination population samples showed significantly higher CD38 enzyme activity in the elderly group than that in the young group ( t=-2.776, P=0.007) and measurement of 30 colorectal cancer patients showed significantly higher CD38 enzyme activity than that in healthy people ( t=-2.572, P=0.012). Conclusion:The established HPLC method for determining CD38 enzymatic activity is characterized by its simplicity, efficiency, accuracy, and reproducibility. This technique serves as a valuable tool for investigating aging and aging-related diseases.

Intracranial aneurysm is a common cerebrovascular disease, which has a high morbidity and mortality after rupture, usually resulting in poor prognosis. Intracranial aneurysms are mainly treated by craniotomy clipping and endovascular embolization, but there is still controversy about whether the unruptured aneurysms with a diameter of <5 mm need intervention. Studies have shown that inflammation plays an important role in the formation, progression, and rupture of intracranial aneurysms. Aspirin and statins can delay the development of intracranial aneurysms and help reduce the risk of rupture through anti-inflammatory. This article reviews the inflammatory mechanism and potential drug therapy of intracranial aneurysms.

Objective 　To evaluate the role of surgical resection on synchronous multiple pulmonary nodules identified difficultly in clinics. Methods 　The clinical data of 97 patients with synchronous multiple pulmonary nodules who received surgical resection between 2012 and 2019 in Hunan Cancer Hospital were retrospectively analyzed. There were 72 males and 25 females, aged 58.1卤9.0 years. Among these patients, there were 78 patients with ipsilateral and 19 patients with bilateral pulmonary nodules. Clinicopathological parameters between main nodules and secondary nodules were evaluated. Perioperative morbidity was also assessed. Results 　The operation was successfully completed on all patients for the ipsilateral and bilateral lesions. Totally, 71.1% of mian lesions was mostly removed by lobectomy, and the completion rate of video-assisted thoracoscopic surgery (VATS) was 69.1% (67/97); 80.4% of secondary lesions were mostly removed by wedge resection, and the completion rate of VATS was 71.1% (69/97). The incidence of grade 3 or higher complications after unilateral or bilateral surgery was 12.8% and 5.3%, respectively. Postoperative pathology confirmed that the main lesions were malignant in 65 patients (67.0%), mainly adenocarcinoma (63.1%), of which 43.1%were in the stage Ⅰ; 32 patients were benign, mainly tuberculoma (56.3%). There were 29 patients of malignant secondary lesions, 67 benign, and 1 both benign and malignant; the pathological agreement rate of primary and secondary lesions was 54.6% (lung cancer metastases in the lung and all the benign). When the primary lesion was malignant with its diameters of <3 cm, 3-<5 cm, 5-7 cm, >7 cm, the metastatic rate of secondary lesions was 42.5%, 15.8%, 20.0%, 0, respectively. When the primary lesion was malignant with lymph node metastasis, the probability of the secondary lesion being a metastatic tumor was higher than that without lymph node metastasis (46.7% vs. 30.0%, P>0.05). When the primary lesion was malignant and the primary and secondary lesions were located in the same lobe, the secondary lesions were more likely to metastasize (54.5%), while when they were located on different lobes on the same side or different sides, the secondary lesions were more likely to be benign (58.1%, 72.7%), and the possibility of metastasis was small ( 32.6%, 9.1%). When the primary lesion was benign and clinical differential diagnosis was difficult, the secondary lesion was benign. Conclusion    For synchronous multiple pulmonary nodules, the diameter of the primary lesion is large, the metastatic rate of secondary lesions tends to decrease. In ipsilateral synchronous multiple pulmonary nodules, especially with node metastasis, the risk of metastatic nodule increases. Bilateral surgical resection does not significantly increase the perioperative morbidity.

Objective:To realize the bacterial distribution and antibiotic resistance in children with severe pneumonia in this region.Methods:A total of 203 children with severe pneumonia diagnosed in Gansu Provincial People′s Hospital from April 2018 to March 2020 were divided into 0-1, 1-3, 3-7 and 7-14 years old groups.Bronchoalveolar lavage fluid was collected for bacterial culture and identification, and antibiotic susceptibility tests were performed.Results:The positive rate of pathogens was 69.5% (141/203), including 72.3% (102 strains) of Gram-negative bacteria and 30.5%(43 strains)of Gram-positive bacteria.The infection rates were highest in 0-1 years old group and the lowest in 7-14 years old group, which were 45.2%(19/42) and 16.9%(10/59), respectively.The infection rates of Haemophilus influenzae, Escherichia coli and Branhamella catarrhalis in the 1-3 years old group were 30.30%(10/33), 33.33% (11/33), and 21.21% (7/33), respectively, which showed significant differences compared with other groups( P<0.05). The infection rate of Streptococcus pneumoniae in the 0-1 years old group was 42.9%(18/42), which was significantly different compared with other groups ( P<0.001). The resistance rate of Haemophilus influenzae to trimethoprim/sulfamethoxazole was 89.5%(34/38), and the Streptococcus pneumoniae to trimethoprim/sulfamethoxazole and tetracycline were both 82.4%(28/34). The highest antibiotic resistance rate of Escherichia coli was 34.6%(9/26), and the Branhamella catarrhalis to clindamycin was 56.3%(9/16). Conclusion:The dominant bacteria for severe pneumonia in children are Haemophilus influenzae, Streptococcus pneumoniae, Escherichia coli and Branhamella catarrhalis.The bacterial infection rate is highest within 1 year old, but gradually decreases with the increase of age.Haemophilus influenzae and Streptococcus pneumoniae have severe resistance to several antibiotics.

ObjectiveTo determine the pathogenic characteristics and genotype of two outbreaks of herpangina in children in Dapeng New District， Shenzhen， in May 2021. MethodsA total of five throat swabs from children in the two outbreaks of herpangina were collected and examined for common enteroviruses by real-time PCR. The VP1 region was further amplified by nested RT-PCR. The CLUSTAL W program in MEGA7 software was used to conduct the alignment and reconstruct a phylogenetic tree. ResultsThe pathogen causing the 2 cluster outbreaks of herpangina was coxsackievirus A4 （CVA4）. The sequences of CVA4 VP1 genes revealed that a nucleotide identity of 92% between the strains in the two outbreaks. The three CVA4 strains isolated in kindergarten A had the closest phylogenetic relationship with that isolated in Shenzhen in 2018（MN840533）， with the nucleotide identity of 98.11%. The two strains in kindergarten B had the closest phylogenetic relationship with CVA4 strain isolated in Sichuan in 2018（MW178763）， with the nucleotide identity of 97.88%. The phylogenetic tree showed that all five CVA4 strains in this study belonged to the C2 genotype. ConclusionThe C2 genotype of CVA4 is the causative agent in both outbreaks of herpangina.

With the dramatically increasing detection rate of ground-glass nodules (GGN), exact understanding and treatment strategy of them has become the hottest issue currently. More and more studies have begun to explore the underlying mechanisms of their indolent characteristics and favorable prognosis from the perspectives of molecular evolution and immune microenvironment. GGN has different dominating gene mutations at different evolutional stages. The pure GGN has a lower tumor mutation burden and genomic instability, while a gradually evolutionary feature of genomic mutation along with the pathological progression can be observed. GGN has less infiltration of immune cells, and they are under the pressure of immune surveillance with weakened immune escape. With the increase of solid components, an inhibitory immune microenvironment is gradually established and immune escape is gradually enhanced, leading to rapid tumor growth. Further exploration of the molecular characteristics of GGN will help to more precisely distinguish these highly heterogeneous lesions, which could be helpful to make personalized treatment plans.