ObjectiveThis study aims to investigate the regulatory effects of microcapsules with different matrix stiffness on the survival, proliferation, adhesion, and osteogenic differentiation of mesenchymal stem cells (MSCs), to further explore the relationship between matrix stiffness and MSCs osteogenic behavior, and to evaluate the in vivo histological responses and osteogenic-related protein expression of the microcapsules using an animal subcutaneous implantation model. MethodsIn this study, three-dimensional carbon nanofiber (CNF)/sodium alginate (SA) microcapsules with different matrix stiffness were constructed. By adjusting the CNF content, three groups of microcapsules with stiffness values of 27.05, 35.90, and 52.40 kPa were prepared, and their physicochemical properties, including morphology, mechanical properties, particle size, chemical structure, and degradation properties, were characterized. The effects of these microcapsules with different stiffness on the survival, adhesion, and osteogenic differentiation of MSCs were evaluated. Live/dead staining was used to observe cell survival and distribution in the microcapsules, while alkaline phosphatase (ALP) activity and the expression of osteopontin (OPN) and osteocalcin (OCN) were examined to assess osteogenic differentiation. Immunofluorescence staining was further performed to observe the expression of Yes-associated protein (YAP) and integrin β1 under different stiffness conditions. In addition, a rat subcutaneous implantation model was used to evaluate the in vivo tissue compatibility of the microcapsules. Histological staining and immunohistochemical analysis were carried out to examine local tissue responses and the expression of osteogenic-related proteins. ResultsThe results showed that all three groups of microcapsules exhibited regular and uniform morphology and similar particle size, and all groups showed good degradability. Except for the differences in mechanical properties, no obvious differences were found in the other physicochemical properties among the three groups. Live/dead staining indicated that all groups of microcapsules had good cytocompatibility. MSCs in the low-stiffness group were mostly round, whereas cells in the medium- and high-stiffness groups showed more obvious spreading. Higher matrix stiffness was more favorable for MSC adhesion, growth, and survival. ALP staining and quantitative analysis showed that ALP activity increased significantly with increasing matrix stiffness. The expression of the osteogenic-related proteins OPN and OCN also increased with increasing stiffness, and the high-stiffness group showed the strongest positive signals. Immunofluorescence results further indicated that YAP and integrin β1 fluorescence signals tended to increase in MSCs with increasing matrix stiffness, suggesting that matrix stiffness may influence MSC osteogenic differentiation through stiffness-related cell adhesion and mechanotransduction. In vivo experiments showed that all three groups of microcapsules exhibited good histocompatibility after subcutaneous implantation, although different degrees of degradation and local tissue response were observed. Immunohistochemical analysis showed that the expression levels of OPN and OCN increased with increasing matrix stiffness, and the high-stiffness group exhibited higher expression of these osteogenic-related proteins. ConclusionCNF endows SA microcapsules with tunable matrix stiffness. While maintaining good biocompatibility, the high-stiffness microenvironment can effectively promote the adhesion and osteogenic differentiation of MSCs, accompanied by increased ALP activity and enhanced OPN and OCN expression. The increased YAP and integrin β1 fluorescence signals in the high-stiffness group further indicate that matrix stiffness is closely related to cellular mechanical responses. These findings provide a three-dimensional cell carrier with potential application value for MSC delivery and bone tissue regeneration based on mechanical microenvironment regulation.

Objective To explore the perfusion distribution of vertebral arteries(VA)in healthy adults,and to observe the impact of VA diameter on perfusion distribution based on territorial arterial spin labeling(t-ASL)technique.Methods A total of 228 healthy adult volunteers were prospectively recruited.t-ASL imaging was utilized to analyze VA perfusion patterns in supratentorial region.Bilateral VA diameters,absolute inter-side VA diameter differences,as well as perfusion volumes(PV)supplied by each VA to supratentorial,cerebellar and their combined were recorded.Pearson correlation analysis was performed to analyze the correlations of VA diameters with PVsupratentorial,PVcerebellar and PVsupratentorial+cerebellar.Results Five patterns of distinct perfusion distribution were identified in supratentorial region,leading to participant stratification into 5 groups.Significant differences of absolute inter-side VA diameter differences was found among groups(P＜0.001).The mean bilateral VA diameter,PVsupratentorial,PVcerebellar and PVsupratentorial+cerebellar was(1.94±0.49)mm,(91.77±49.34)cm3,(74.72±27.68)cm3 and(166.49±59.53)cm3,respectively.VA diameter showed low-moderate positive correlations with PVsupratentorial,PVcerebellar and PVsupratentorial+cerebellar(r=0.483,0.179,0.484,all P＜0.001).Conclusion Perfusion imaging of t-ASL could visualize various perfusion distribution forms of VA in supratentorial region.Diameter of VA had certain impact on both spatial distribution characteristics and quantitative perfusion volumes.

Objective To investigate the knowledge and attitude of clinical nurses on Panvascular Disease(PVD)care in Zhejiang Province,and to analyze the influencing factors,so as to provide references for constructing a systematic PVD training program.Methods The convenient sampling method was used to investigate 454 nurses from 36 general hospitals in 11 prefectures and cities in Zhejiang Province in June 2024 with a self-designed questionnaire on general information and a questionnaire on PVD nursing knowledge and attitude questionnaire,and multiple stepwise linear regression was used to analyze the impact factors.Results 452 valid questionnaires were collected and the valid recovery rate was 99.56％.Nurses scored(8.57±2.02)in knowledge dimension,(60.82±6.45)in attitude dimension.The results of multiple stepwise linear regression analysis showed that hospital grade was the factor influencing the score of nurses'knowledge towards PVD nursing in Zhejiang Province(P＜0.05).Hospital grade,whether they had received systematic knowledge training related to PVD were the factors influencing the score of nurses'attitude towards PVD nursing(P＜0.05).Conclusion Nurses in Zhejiang Province are positive about PVD nursing,but their knowledge level needs to be improved.Nursing managers should pay full attention to the training intention of clinical nurses,establish a PVD knowledge training system,and strengthen the theoretical and practical training of clinical nurses on PVD nursing,so as to promote the development of PVD nursing work.

The " no-accompany" service is an important measure to meet the patients′ needs for comprehensive, continuous, and high-quality diversified care and to enhance their medical experience. Starting in March 2024, the Second Affiliated Hospital of Zhejiang University School of Medicine launched a pilot " no-accompany" service in the breast surgery and dermatology wards. The hospital established a working team led by the nursing department and involving multiple administrative departments such as the medical affairs department, medical insurance office, and logistics management department. And the hospital has successfully advanced the implementation of the " no-accompany" ward program by establishing a multi-department collaborative management mechanism, standardizing the management of medical caregivers, developing a tiered fee scheme for the " no-accompany" service, and improving relevant service measures. The acceptance of the " no-accompany" service by patients increased from (3.93±0.83) in the second quarter of 2024 to (4.69±0.59) in the fourth quarter, and overall satisfaction rose from (4.18±0.73) to (4.50±0.54) (all P<0.001). The job competency of medical caregivers also improved significantly from (64.80±3.49) before starting work in March 2024 to (94.00±2.40) in the fourth quarter ( P<0.001). These findings provide a reference for the implementation of " no-accompany" services in public hospitals.

Objective To investigate the current status and barriers to implementing non-accompanied care services in medical institutions in Zhejiang Province,and to provide a basis for the improvement of standardized management of such services.Methods A self-designed questionnaire was conducted from September to October 2024 among all secondary and tertiary medical institutions in Zhejiang Province to assess the implementation status and barriers to non-accompanied care services.Results A total of 397 questionnaires were distributed,with 389 valid responses,yielding a valid response rate of 97.98％.Non-accompanied care services were implemented in 118 institutions(30.33％).Among these,90 institutions(76.27％)had established management systems for non-accompanied wards;71 institutions(60.17％)had a medical nursing assistant-to-bed ratio lower than 1∶5;41 institu-tions(34.75％)provided tiered training for medical nursing assistants;93 institutions(78.81％)required patients to bear the full cost of the service.Compared with secondary medical institutions,tertiary medical institutions have more complete management system for non-accompanied care services.The main obstacles hindering the development of non-accompanied care services include an imperfect management system for non-accompanied wards,a shortage of medical nursing assistants,a lack of standardized training for such assistants,inconsistent charging standards,and low acceptance among patients and their families.Conclusion The promotion of non-accompanied care services in medical institutions in Zhejiang Province has achieved initial success.However,challenges persist,including incomplete management systems,uneven development across hospital tiers,and imperfect charging mechanisms.It is recommended that relevant authorities strengthen policy support,enhance standardized training for healthcare nursing assistants,refine cost-sharing mechanisms,and improve the quality and sustainability of non-accompanied care services through multi-party collaboration.

Objective:To investigate the efficacy of posterior cranial fossa in- situ floating bone flap osteotomy in the treatment of syndromic craniosynostosis(SCS). Methods:The clinical data of SCS children who underwent posterior cranial fossa in- situ floating bone flap osteotomy at the Shanghai Children’s Medical Center, Shanghai Jiao Tong University School of Medicine from April 2020 to August 2022 were retrospectively analyzed. The surgical procedures were as follows. The occipital bone was cut into several mosaic bone flaps of varying sizes, without peeling it off the dura, which were left as a small free floating bone flap. The anteroposterior cranial diameter, cranial height, intracranial volume, and degree of improvement in tonsillar herniation and hydrocephalus of the children (hydrocephalus quantification was performed using the ratio of the ventricular diameter to the biparietal diameter)were measured preoperatively, 7 days postoperatively, 3 months postoperatively, and at the last follow-up (at least 12 months after the operation) to evaluate the surgical outcomes. The measurement data of normal distribution were expressed as Mean±SD. The paired t-test was used for comparison within the repeated measurement data groups at the two time points. One-way repeated measures analysis of variance (ANOVA) was performed on the repeated measurement data at multiple time points, and pairwise comparisons in post hoc tests were corrected using the Bonferroni method. Results:A total of 17 pediatric patients with SCS were included, comprising 10 males and 7 females, with ages ranging from 4 to 18 months (mean age: 9.5 months). Among them, 12 patients were complicated with Chiari malformation and hydrocephalus (1 severe case, 8 moderate cases, and 3 mild cases). Postoperative follow-up lasted 12 to 35 months, with an average of 17 months. After surgery, the posterior cranial appearance of the children was enlarged, with increased convexity of the occiput and a full contour. At the last follow-up, the middle cranial height [(107.80±10.72) mm vs. (102.82±10.09) mm, P<0.05], the posterior cranial height [(124.91±10.40) mm vs. (107.58±13.46) mm, P<0.01] and anteroposterior diameter [(153.30±11.26) mm vs. (123.64±17.44) mm, P<0.01] as well as the intracranial volume [(1 317.92±225.77) cm 3 vs. (1 014.93±231.81) cm 3,P<0.01] were increased compared with the preoperative period, and the average improvement rate of intracranial volume was 37.0% (18.1%-79.2%). Among the 12 cases of tonsillar herniation, 7 cases had improvement. Moreover, all the 12 cases of hydrocephalus witnessed a mitigation in severity, from (46.33±9.34)% preoperatively to (35.24±9.88)% postoperatively, with a statistically significant difference ( P<0.01). Conclusion:Posterior cranial fossa in- situ floating bone flap osteotomy can effectively improve the appearance of patients with SCS, increase the intracranial volume, relieve the degree of hydrocephalus, and reduce the intracranial pressure.

Nitrogen oxides(NOx=NO+NO2)are important precursors of ozone(O3),and NOx and its oxides together constitute reactive nitrogen oxides(NOy)in the atmosphere.A comprehensive understanding of the total NOy level in the atmosphere is of great significance for a deeper understanding of the atmospheric nitrogen cycle and oxidation,as well as for formulating strategies for air pollution prevention and control.In this work,a thermal decomposition-broadband cavity enhanced absorption spectroscopy(TD-BBCEAS)technique for online measurement of total NOy in the atmosphere was developed.With this method,the NOy was efficiently converted into NO2,and the total NOy concentration in the atmosphere was indirectly obtained by measuring NO2.Focusing on the key factors affecting the measurement of total NOy,the influence of NO titration efficiency and other NOy component TD efficiency on measurement accuracy was emphasized.By changing the oxygen(O2)flow rate through the mercury lamp to alter the O3 concentration for titrating NO,the conversion efficiency of NO was evaluated.At O2 flow rate of 6 mL/min,the conversion efficiency of NO was greater than 99%.TD efficiency testing and analysis on NO2,peroxyacetyl nitrate(PAN),nitric acid(HNO3),and nitrous acid(HONO),which account for a large proportion of atmospheric NOy components,was carried out using 680℃as the optimal TD temperature for efficient conversion of NOy.With NO and HONO sample gases as typical verification gases,the conversion efficiency of NOy and the accuracy of NOy measurement by TD-BBCEAS system were verified by switching the on and off modes of mercury lamp and TD device.At integration time of 60 s,the detection limit of the system for NOy was 2.83×1010 molecules/cm3(60 s,2σ).A comparative measurement of actual atmospheric NOy was conducted between the TD-BBCEAS system and the NOy analyzer.The observation results showed a correlation coefficient(R2)of 0.98 and a slope of 0.93,further verifying the feasibility and accuracy of applying the TD-BBCEAS system to measurement of total NOy.

BACKGROUND:The authors found that when the bilateral percutaneous vertebral augmentation is used to treat osteoporotic vertebral compression fractures with a total bone cement injection of 4 mL or more,different distribution patterns were usually presented on the X-rays;however,there were few reports addressing the effects of these patterns of bone cement distribution on the biomechanical properties of fractural vertebrae. OBJECTIVE:To further explore the biomechanical effects of different bone cement filling doses and distribution patterns on biomechanics of the fractural vertebrae using the finite element method. METHODS:The L1-L3 finite element models of osteoporosis were established,and the vertebral compression fractures were simulated in L2.Four distribution patterns bilateral partial fusion(FH type),full fusion(FO type),symmetrical separation(SA type),and asymmetric segregation(SN type)were simulated in 4 and 6 mL injections in the osteoporotic vertebral compression fracture models,respectively,and a total of nine sets of models were obtained.These models were solved under the same boundary conditions and compared with the stress and displacement of the L2 fractural vertebra. RESULTS AND CONCLUSION:(1)The maximum stresses of the nine groups of models were concentrated in the L2 fractural area,and the maximum stress and maximum displacement of each filling model were lower than in the osteoporotic vertebral compression fracture model,indicating the effectiveness of bone cement filling in the treatment of osteoporotic vertebral compression fracture.(2)Compared with 4 mL bone cement filling,6 mL bone cement filling could significantly reduce the stress of fractured vertebrae and enhance the strength of fractured vertebrae while improving the stability of fractured vertebrae.(3)In the same state of movement,the FH type stress was the least,followed by the SA type,both of which were close.FO type stress was the largest,especially in the lateral bend,which might be associated with its cluster shape resulting in the concentration of lateral stress.In the aspect of displacement,FH type was the least and FO type was the largest.(4)The results show that increased dose of bone cement injection reduces fractural vertebral stress and improves stability,but increases the risk of leakage.Bilateral symmetrical dispersed bone cement(FH type,SA type)is superior in restoring vertebral strength and stability than full fusion(FO type),asymmetric separated(SN type)bone cement.Therefore,when clinically performing bilateral percutaneous vertebral augmentation treatment of osteoporotic vertebral compression fractures,the bilateral symmetric dispersions of the distribution are first guaranteed;priority is recommended for FH type distribution,for appropriate stress stimulation and best stability.

PURPOSE: To investigate the pathological changes of the synovium in mice with post-traumatic osteoarthritis (PTOA) treated with 4-octyl itaconate (4-OI) and evaluate the therapeutic effects of 4-OI. METHODS: In the phenotypic validation experiment, the mice were randomly divided into 3 groups: wild-type (WT) group, sham group, and destabilization of the medial meniscus (DMM) group. Through MRI, micro-CT, and histological analysis, it was determined that the DMM surgery induced a mouse PTOA model with significant signs of synovitis. At 12 weeks post-DMM surgery, synovial tissues from the DMM group and WT group mice were collected for ribonucleic acid sequencing analysis. In the 4-OI treatment experiment, mice were randomly divided into the sham group, DMM group, DMM + 4-OI (50 mg/kg) group, and DMM + 4-OI (100 mg/kg) group. Von Frey tests and open field tests were conducted at intervals during the 12 weeks following the DMM surgery. After 12 weeks of surgery, the efficacy of 4-OI treatment on PTOA in mice was evaluated using MRI, micro-CT, histological analysis, and quantitative real-time polymerase chain reaction. Finally, we utilized network pharmacology analysis to predict the mechanism of 4-OI in treating PTOA synovitis and conducted preliminary validation. Statistical analysis was performed using one-way ANOVA and the Kruskal-Wallis test. Difference was considered statistically significant at p < 0.05. RESULTS: The DMM surgery effectively induced a PTOA mouse model, which displayed significant symptoms of synovitis. These symptoms included a notable increase in both the number of calcified tissues and osteophytes (p < 0.001), an enlargement of the calcified meniscus and synovial tissue volume (p < 0.001), and thickening of the synovial lining layer attributable to M1 macrophage accumulation (p = 0.035). Additionally, we observed elevated histological scores for synovitis (p < 0.001). Treatment with 4-OI inhibited the thickening of M1 macrophages in the synovial lining layer of PTOA mice (p < 0.001) and reduced fibrosis in the synovial stroma (p = 0.004). Furthermore, it reduced the histological scores of knee synovitis in PTOA mice (p = 0.006) and improved the inflammatory microenvironment associated with synovitis. Consequently, this treatment alleviated pain in PTOA mice (p < 0.001) and reduced spontaneous activity (p = 0.003). Bioinformatics and network pharmacology analyses indicated that 4-OI may exert its therapeutic effects by inhibiting the differentiation of synovial Th17 cells. Specifically, compared to the lipopolysaccharide stimulation group, 4-OI reduced the levels of positive regulatory factors of Th17 cell differentiation (IL-1: p < 0.001, IL-6: p < 0.001), key effector molecules (IL-17A: p < 0.001, IL-17F: p = 0.004), and downstream effector molecules in the IL-17 signaling pathway (CCL2: p < 0.001, MMP13: p < 0.001). CONCLUSION 4-OI is effective in inhibiting synovitis in PTOA, thereby alleviating the associated painful symptoms.
Animals ; Synovitis/etiology* ; Mice ; Osteoarthritis/etiology* ; Disease Models, Animal ; Male ; Succinates/pharmacology* ; Mice, Inbred C57BL ; X-Ray Microtomography

The genome tagging project (GTP) plays a pivotal role in addressing a critical gap in the understanding of protein functions. Within this framework, we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411 (HA-tagged Ssp411) mouse model. This model is instrumental in probing the expression and function of Ssp411. Our research revealed that Ssp411 is expressed in the round spermatids, elongating spermatids, elongated spermatids, and epididymal spermatozoa. The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis. Nevertheless, rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions. Ssp411 is not detectable in metaphase II (MII) oocytes, zygotes, or 2-cell stage embryos, highlighting its intricate role in early embryonic development. These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP, fostering groundbreaking advancements in the fields of spermiogenesis and reproductive biology.
Animals ; Female ; Humans ; Male ; Mice ; Spermatids/metabolism* ; Spermatogenesis/physiology* ; Spermatozoa/metabolism* ; Thioredoxins/genetics*