BACKGROUND:Ursolic acid can promote the directed differentiation of bone marrow mesenchymal stem cells into osteoblasts.However,there are few reports on whether ursolic acid has osteogenic effect on human periodontal ligament stem cells. OBJECTIVE:To investigate the effect of ursolic acid on proliferation and osteogenic differentiation of human periodontal ligament stem cells. METHODS:The human periodontal ligament stem cells were isolated and cultured.Passage 3 cells were selected and treated with ordinary medium containing different concentrations(0,1,2,4,6,8 μmol/L)of ursolic acid.After intervention for 1,3,5,7 days,the cell proliferation was detected by CCK-8 assay and the appropriate intervention concentration was screened.Passage 3 human periodontal ligament stem cells were treated with osteogenic induction solution containing 0,1,2,4 μmol/L ursolic acid,respectively.After 7 days of intervention,the mRNA expressions of alkaline phosphatase,Runx2,and osteocalcin were detected by qRT-PCR.After 14 days of intervention,the formation of mineralized nodules was observed by alizarin red staining.Passage 3 human periodontal ligament stem cells were taken and the control group was added with osteogenic induction solution;the ursolic acid group and the antagonist group were added with osteogenic induction solution containing ursolic acid(2 μmol/L)and the bone morphogenetic protein signaling pathway antagonist Noggin,respectively.The ursolic acid+antagonist group was added with osteogenic induction solution containing ursolic acid(2 μmol/L)and Noggin,the inhibitor of bone morphogenetic protein signaling pathway,and cultured for 7 days.qRT-PCR and western blot assay were used to detect the mRNA and protein expressions of bone morphogenetic protein 2,Smad1,osteopontin,and Runx2. RESULTS AND CONCLUSION:(1)1,2,4 μmol/L ursolic acid could promote the proliferation of human periodontal ligament stem cells.6,8 μmol/L ursolic acid could inhibit the proliferation of human periodontal ligament stem cells,and 1,2,4 μmol/L ursolic acid was selected to intervene in subsequent experiments.(2)Compared with 0 μmol/L,1,2,4 μmol/L ursolic acid could promote the expression of alkaline phosphatase,Runx2,and osteocalcin mRNA and the formation of mineralized nodules(P<0.05),and the effect of 2 μmol/L ursolic acid was the most significant.(3)Compared with the control group,the mRNA and protein expressions of bone morphogenetic protein 2,Smad1,osteopontin,and Runx2 in the ursolic acid group were increased(P<0.05),while mRNA and protein expressions of the above indexes were decreased in the antagonist group(P<0.05).Compared with the ursolic acid group,mRNA and protein expressions of above indexes were decreased in ursolic acid+antagonist group(P<0.05).(4)The results indicate that ursolic acid promotes osteogenic differentiation of human periodontal ligament stem cells through bone morphogenetic protein signaling pathway.

Objective To investigate causal relationship between gut microbiota and stroke by using Mendelian randomization(MR).Methods The gut microbiota genome wide association study(GWAS)data of MiBioGen study in public database(n=13 266)and stroke GWAS data of IEU Open GWAS were used.Independent genetic loci significantly correlated with the relative abundance of gut microbiota at genus level were used for instrumental variables(Ⅳs)according to preset thresholds.The inverse variance weighted method was used to analyse,results were evaluted according to odds ratio(OR)of effect index and 95％confidence interval(CI).The stability and reliability of the results were verified by leave one out,heterogeneity test and horizontal gene pleiotropy test.Results The rise of abundance of LachnospiraceaeFCS020group genera,Lachnospiraceae NK4A136group genera,RuminococcaceaeUCG004 genera and Intestinimonas genera,could reduce the risk of stroke,the OR(95％CI)were 0.914(0.845-0.988),0.909(0.838-0.985),0.919(0.849-0.995),0.924(0.866-0.987),respectively.The increase of abundance of Allisonella genera,Gordonibacter genera,Paraprevotella genera and Streptococcus genera can increase the risk of stroke,and the OR(95％CI)were 1.103(1.039-1.171),1.056(1.009-1.105),1.078(1.013-1.147),1.110(1.020-1.209),respectively.Leave one out showed that the results were stable,there were no IVs that had a strong influence on the results,and the influence of heterogeneity and horizontal gene pleiotropy on the estimation of causal effect could be eliminated.Conclusion The levels of Lachnospiraceae FCS020group genera,LachnospiraceaeNK4A136group genera,RuminococcaceaeUCG004 genera and Intestinimonas genera protect factors of brain stroke,with the increase of bacterial abundance,the incidence of stroke may be reduced.On the contrary,the levels of Allisonella genera,Gordonibacter genera,Paraprevotella genera and Streptococcus genera are positively correlated with stroke occurrence.Probiotics in the treatment of stroke may become the trend of future clinical or mechanism research.

Objective To investigate the current status and awareness of pharmaceutical services in hospitals in Guizhou province and to provide a reference for exploring and carrying out pharmaceutical service fees.Methods The questionnaire was designed by the"wjx.cn"website.Three kinds of questionnaires were designed for pharmacists,doctors,nurses,and patients as the research objects,with corresponding differences in some questions,and promoted on WeChat,Dingxiangyuan,and other network platforms.Results A total of 655 questionnaires were collected,and 639 valid questionnaires were recovered,with an effective recovery rate of 97.56%.324 pharmacists(50.70%),82 doctors and nurses(12.83%),233 patients(36.46%)were surveyed.The average approval score of these three groups of respondents on pharmaceutical service fees was 4.67,4.23,and 4.22,respectively(full score:5).Conclusions Overall,pharmacists'professional services have received support from medical staff and patients.However,patients'pharmaceutical service projects currently focus on dispensing services.The recognition of pharmacists'work and the public's awareness of pharmaceutical services can be improved by enhancing the professional ability of pharmacists,strengthening publicity and guidance,and exploring"Internet+pharmaceutical services",etc.,to promote the sustainable development of pharmaceutical services.

OBJECTIVES: Acute ischemic stroke (AIS) is one of the main causes of disability in middle-aged and elderly people, and early activity plays an important role in functional recovery. This study aims to understand the factors that affect the implementation of early activity in patients with AIS and to provide reference for promoting early activity implementation and developing intervention strategies for AIS patients. METHODS: Using purposive sampling, 19 AIS patients and their caregivers who visited at Stroke Center in the Third Xiangya Hospital of Central South University and the Third Hospital of Changsha from June to December 2021, as well as 19 medical staff, hospital administrators, or community workers providing medical health services to stroke patients, were selected as interviewes. A semi-structured interview was conducted based on the social ecological theory model, and the Colaizzi seven-step method was used to analyze the interview data. RESULTS: According to qualitative interview results, the factors affecting early activity in AIS patients were summarized into 4 themes and 12 sub-themes: medical staff factors (insufficient knowledge and skills, insufficient knowledge of early activity, unclear division of responsibilities), patient factors (severity of the disease, lack of knowledge, psychological pressure, fear of falling), social environmental factors (lack of social support, shortage of human resources and rehabilitation equipment, insufficient medical insurance support), and evidence and norms (the evidence for early activity needs improvement, lack of standardized early activity procedures). CONCLUSIONS Early activity in AIS patients is impacted by factors at multiple levels, including medical staff, patients, social environment, and evidence and norms. Developing comprehensive intervention strategies to address these factors can promote early activity implementation in AIS patients.
Aged ; Middle Aged ; Humans ; Ischemic Stroke ; Accidental Falls ; Fear ; Social Environment ; Stroke

In growing children, growth plate cartilage has limited self-repair ability upon fracture injury always leading to limb growth arrest. Interestingly, one type of fracture injuries within the growth plate achieve amazing self-healing, however, the mechanism is unclear. Using this type of fracture mouse model, we discovered the activation of Hedgehog (Hh) signaling in the injured growth plate, which could activate chondrocytes in growth plate and promote cartilage repair. Primary cilia are the central transduction mediator of Hh signaling. Notably, ciliary Hh-Smo-Gli signaling pathways were enriched in the growth plate during development. Moreover, chondrocytes in resting and proliferating zone were dynamically ciliated during growth plate repair. Furthermore, conditional deletion of the ciliary core gene Ift140 in cartilage disrupted cilia-mediated Hh signaling in growth plate. More importantly, activating ciliary Hh signaling by Smoothened agonist (SAG) significantly accelerated growth plate repair after injury. In sum, primary cilia mediate Hh signaling induced the activation of stem/progenitor chondrocytes and growth plate repair after fracture injury.
Mice ; Animals ; Hedgehog Proteins/genetics* ; Receptors, G-Protein-Coupled/metabolism* ; Cilia/metabolism* ; Cartilage/metabolism* ; Regeneration

The mechanisms underlying autophagic defects in nonalcoholic steatohepatitis (NASH) remain largely unknown. We aimed to elucidate the roles of hepatic cyclooxygenase 1 (COX1) in autophagy and the pathogenesis of diet-induced steatohepatitis in mice. Human nonalcoholic fatty liver disease (NAFLD) liver samples were used to examine the protein expression of COX1 and the level of autophagy. Cox1^Δhepa mice and their wildtype littermates were generated and fed with 3 different NASH models. We found that hepatic COX1 expression was increased in patients with NASH and diet-induced NASH mice models accompanied by impaired autophagy. COX1 was required for basal autophagy in hepatocytes and liver specific COX1 deletion exacerbated steatohepatitis by inhibiting autophagy. Mechanistically, COX1 directly interacted with WD repeat domain, phosphoinositide interacting 2 (WIPI2), which was crucial for autophagosome maturation. Adeno-associated virus (AAV)-mediated rescue of WIPI2 reversed the impaired autophagic flux and improved NASH phenotypes in Cox1^Δhepa mice, indicating that COX1 deletion-mediated steatohepatitis was partially dependent on WIPI2-mediated autophagy. In conclusion, we demonstrated a novel role of COX1 in hepatic autophagy that protected against NASH by interacting with WIPI2. Targeting the COX1-WIPI2 axis may be a novel therapeutic strategy for NASH.

Objective:To understand the current situation of early mobilization in elder patients with acute ischemic stroke, and to analyze the influencing factors of early mobilization in patients with different degrees of neurological deficits, aiming to provide evidence for promoting early mobilization in patients with stroke and for formulating intervention strategies.Methods:Patients with acute ischemic stroke admitted to the stroke centers of two class A tertiary hospitals in Hunan Province between January and December 2021 were enrolled.The degree of neurological impairment, activities of daily living and muscle strength were evaluated within 24 hours after admission, and patients' performance of early mobilization was recorded.Based on whether patients performed out-of-bed activities such as sitting at the bedside, standing or walking, transferring from bed to chair within 48 hours after admission, they were divided into an early ambulation group and a non-early ambulation group.Univariate analysis and Logistic regression analysis were then used to examine the influencing factors of early ambulation in patients with mild to moderate neurological deficits.Results:Among 257 patients with acute ischemic stroke, 60.7%(156 cases)conducted early mobilization.The rate of early mobilization for patients with moderate neurological deficits was only 12.9%(12/193), significantly lower than that for patients with mild neurological deficits(87.8%, 144/164)( χ2=139.571, P<0.001).Univariate analysis showed that there were significantly differences in the level of activities of daily living, stroke history, strength of the upper and lower limbs of the affected side between the early and non-early ambulation groups with moderate neurological impairment(all P<0.05), and there were significant differences in the level of activities of daily living between the early ambulation group and the non-early ambulation group in patients with mild neurological impairment( χ2=11.907, P<0.001).Logistic regression analysis showed that requiring assistance in daily life( OR=4.189, 95% CI: 1.724-10.668, P=0.002), muscle strength of the affected lower extremity ≤ grade 3( OR=9.605, 95% CI: 2.627-35.113, P=0.001)and moderate neurological deficits( OR=8.585, 95% CI: 3.216-22.913, P<0.001)were the risk factors of early mobilization in stroke patients. Conclusions:The rate of stroke patients with moderate neurological impairment performing early mobilization is low within 48 hours after admission.Activities of daily living ability, limb muscle strength and degrees of neurological deficits are influencing factors of early mobilization in stroke patients.

Currently, cardiovascular disease has become a major threat to the health of Chinese residents, and prevention and control work is urgent. Self-determination theory is a widely applied theory of behavioral change, extensively used in the field of patient' rehabilitation. This article reviews the content, relevant theoretical models, motivation measurement methods, application status and influencing factors of self-determination theory in the rehabilitation of cardiovascular disease patients, so as to provide a basis for promoting the transition of cardiovascular disease patients to healthy lifestyles and maintaining healthy behavior.

BACKGROUND: The study aims to correlate osteogenesis with autophagy during the mineralization induction of MC3T3-e1 through exploring the expression of runt-related transcription factor 2 (RUNX2)/lysosomal-associated transmembrane protein 5 (LAMPT5). METHODS: The induction of mineralization in MC3T3-e1 was followed by detecting the expressions of osteogenesisrelated indexes such as RUNX2, alkaline phosphatase (ALP), osteocalcin (OCN), and LAPTM5 using RT-qPCR and Western blot from 0 to 14 days. Transmission electron microscope was utilised in visualizing the alterations of autophagosomes, which was followed by immunofluorescence detecting the subcellular localization of autophagy-related index sequestosome 1 (P62) and microtubule-associated protein 1 light 3 (LC3) protein and scrutinising the expression of P62 mRNA and P62 and LC3 proteins. RESULTS: Induction of MC3T3-e1 mineralization demonstrated an increased expression of osteogenesis-related indicators such as RUNX2, ALP, OCN, and LAPTM5 (p < 0.05), as evident from the results of RT-qPCR and Western blot. Meanwhile, the expression of autophagosomes increased one day after mineralization induction and then experienced a gradual decline, and enhanced expression of LC3 protein was noted on days 1–2 of mineralization induction but was then followed by a corresponding reduce. In contrast, a continuous increase was reported in the expression of P62 mRNA and protein, respectively (p < 0.05). Up- and down-regulating RUNX2/LAPTM5 expression alone confirmed the aforementioned results. CONCLUSION It was therefore proposed that RUNX2 may be responsible for an early increase and then a gradual decrease in LAPTM5-mediated autophagy through the regulation of its high expression. Meanwhile, increased LAPTM5 expression in osteogenic mineralization presumed that RUNX2/LAPTM5 promoted autophagy and osteogenic expression, which may play a bridging role in the regulation of autophagy and osteogenesis.

Objective:To assess the impact of integrated curriculum (vertically integrated curriculum system) and traditional curriculum system on the perceptions of post competency of undergraduate medical students.Methods:Bibliographic retrieval method and expert consultation were used to compile the questionnaire of perceptions on the post competency of undergraduate medical students. The questionnaire was conducted online to survey the undergraduate medical students from Shanghai Jiao Tong University School of Medicine during March to April, 2019. SPSS 20.0 software was applied for data analysis, chi-square test was used to analyze the classroom teaching forms of medical undergraduates who received different curriculum systems, and their cognition of the doctor post competency and cognitive pathways, and finally the top 10 important competencies were compared.Results:A total of 200 questionnaires were distributed and 167 were recovered, with a recovery rate of 83.5%. Workshop learning was more common in the vertically integrated undergraduate curriculum. Compared with Chinese traditional curriculum, the medical students taught by the vertically integrated undergraduate curriculum were much better informed about competency ( P<0.05). The access to knowledge of competency was also statistically different between two groups of students ( P<0.05). Conclusion:The vertically integrated curriculum is more efficient in helping undergraduate medical students to acquire knowledge about competency.